method for treating dermatological skin disease consisting of eczema, psoriasis, dermatitis, ulcers, shingles, rashes, bedsores, cold sores, blisters, boils, herpes, acne, pimples, warts, and a combination thereof.

[Ignent]

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actual pg lnk
http://patft.uspto.gov/netacgi/nph-Parser?Sect2=PTO1&Sect2=HITOFF&p=1&u=%2Fnetahtml%2FPTO%2Fsearch-bool.html&r=1&f=G&l=50&d=PALL&RefSrch=yes&Query=PN%2F5676977


Subsequent to the filing of the aforementioned patent,

further testing revealed complete 100% destruction of the

AIDS virus in vitro at 20 PPM,

and the fact that said devices were harmless

when ingested and inhaled, being non-toxic.



See pg…

http://patft.uspto.gov/netacgi/nph-Parser?Sect2=PTO1&Sect2=HITOFF&p=1&u=%2Fnetahtml%2FPTO%2Fsearch-bool.html&r=1&f=G&l=50&d=PALL&RefSrch=yes&Query=PN%2F5676977




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United States Patent
5,676,977
Antelman
October 14, 1997
Method of curing AIDS with tetrasilver tetroxide molecular crystal
devices

Abstract

The diamagnetic semiconducting molecular crystal tetrasilver tetroxide
(Ag.sub.4 O.sub.4) is utilized for destroying the AIDS virus,
destroying AIDS synergistic pathogens and immunity suppressing
moieties (ISM) in humans. A single intravenous injection of the
devices is all that is required for efficacy at levels of about 40 PPM
of human blood. The device molecular crystal contains two mono and two
trivalent silver ions capable of "firing" electrons capable of
electrocuting the AIDS virus, pathogens and ISM. When administered
into the bloodstream, the device electrons will be triggered by
pathogens, a proliferating virus and ISM, and when fired will
simultaneously trigger a redox chelation mechanism resulting in
divalent silver moieties which chelate and bind active sites of the
entities destroying them. The devices are completely non-toxic.
However, they put stress on the liver causing hepatomegaly, but there
is no loss of liver function.

Inventors: Antelman; Marvin S. (Rehovot, IL)
Assignee: Antelman Technologies Ltd. (Providence, RI)
Appl. No.: 08/658,955
Filed: May 31, 1996
Related U.S. Patent Documents

Application Number
Filing Date
Patent Number Issue Date
310859 Sep., 1994

Current U.S. Class: 424/618 ; 514/495
Current International Class: A61K 33/38 (20060101); A61K 033/38 ()
Field of Search: 424/618 514/495
References Cited [Referenced By]
U.S. Patent Documents

4415565 November 1983 Wysor
4915955 April 1990 Gomori
4952411 August 1990 Fox, Jr. et al.
5073382 December 1991 Antelman
5078902 January 1992 Antelman
5089275 February 1992 Antelman
5211855 May 1993 Antelman
5223149 June 1993 Antelman
5336499 August 1994 Antelman
5571520 November 1996 Antelman

Other References

"Is The AIDS Virus A Science Fiction?"
by Peter H. Duesberg and Bryan J. Ellison, Policy Review, Summer 1990,
pp. 40-51..

Primary Examiner: Hulina; Amy
Attorney, Agent or Firm: Salter & Michaelson
Parent Case Text

This application is a continuation-in-part of patent application Ser.
No. 08/310,859 filed Sep. 22, 1994, now abandoned.

Claims
What is claimed is:

1. A method of treating AIDS-afflicted humans comprising injecting a
multitude of tetrasilver tetroxide molecular crystals into the
bloodstream of the human subject.

2. A method for increasing white blood cell counts in AIDS-afflicted
humans comprising injecting a multitude of tetrasilver tetroxide
molecular crystals into the bloodstream of the human subject.

3. Methods of treating AIDS-affilicted humans according to claims 1-2
where the concentration of said molecular crystals is approximately 40
PPM of the total blood weight of the human subject.
Description


BACKGROUND OF THE INVENTION

The present invention relates to the employment of molecular crystals
as anti-AIDS devices, but more particularly to the molecular crystal
semiconductor tetrasilver tetroxide Ag.sub.4 O.sub.4 which has two
monovalent and two trivalent silver ions per molecule, and which
through this structural configuration enables intermolecular electron
transfer capable of killing viruses and binding them to the resulting
silver entity so that a single intravenous injection will completely
obliterate acquired immune deficiency syndrome (AIDS) in humans.
Furthermore, said devices are capable of killing pathogens and purging
the bloodstream of immune suppressing moieties (ISM) whether or not
created by the AIDS virus (HIV); so as to restore the immune system.

The present invention is based on concepts previously elucidated in
applicant's U.S. Pat. No. 5,336,499 which discloses the destruction
and inhibition of bacteria, algae and the AIDS virus in nutrient life
supporting systems by using said silver oxide devices. Example 3 of
said patent discloses that 18 PPM of said crystal devices could
totally suppress the AIDS virus (page 6, line 5). Subsequent to the
filing of the aforementioned patent, further testing revealed complete
100% destruction of the AIDS virus in vitro at 20 PPM, and the fact
that said devices were harmless when ingested and inhaled, being non-
toxic.

Encouraged by these evaluations and successes, applicant obtained
permission to evaluate the crystals in vitro against murine acquired
immune deficiency syndrome (MAIDS). Only one facility in the State of
Israel is licensed for these evaluations, namely, the Kaplan Hospital
in Rehovot, Israel, which is affiliated with the Hebrew University-
Hadassah Medical School where said evaluations were done.

The initial evaluations entailed experimenting with various silver
moieties cited in applicant's aforementioned patent, concentrations,
non-reactive buffers and modes of administration. After about 18
months of judicious efforts and initial failures, success was finally
achieved in destroying the MAIDS virus in C57BL mice with a single
intravenous injection. The results of this test program comprise
Example 5 of U.S. Pat. No. 5,336,499. After success with mice, the
inventor was able to test the efficacy of said devices on two select
etiological groups of terminal AIDS patients in a clinic in
Tegucigalpa, Honduras, Central America.

The AIDS patients comprised the etiological subgroups, Candidiasis and
Wasting Syndrome. Current indicator diseases for diagnosing AIDS which
have been expanded by the CDC, fall into the following five major
categories with the approximate percent distribution among AIDS
patients:

______________________________________ 1. P. carinii pneumonia 51% 2.
Wasting syndrome 19% 3. Candidiasis 13% 4. Kaposi's sarcoma 11% 5.
Dementia 6% ______________________________________

This invention concerns itself with the treatment and cure of
candidiasis and wasting syndrome AIDS patients with Tetrasil*. These
two groups account for approximately one third of AIDS cases.

Stedman's Medical Dictionary (Williams & Wilken's 26th Ed., 1995)
defines wasting syndrome "as a condition of 10% weight loss in
conjunction with diarrhea or fever . . . Associated with AIDS (p.
1744)."

OBJECTS OF THE INVENTION

The main object of the invention is to provide for a molecular scale
device of a single tetrasilver tetroxide crystalline molecule capable
of restoring the immunity of AIDS afflicted humans of the two AIDS
etiological subgroups, candidiasis and wasting syndrome.

Another object of the invention is to provide for immunity restoration
in said AIDS afflicted humans through a single injection.

Another object of this invention is to destroy ISM in humans
manifesting AIDS diseases of said AIDS etiological subgroups
irrespective as to whether said ISM was HIV induced, since it is known
that humans may manifest AIDS and still be HIV negative, and thus
restore the immune system in said humans.

Another object of this invention is to destroy the AIDS virus when
present in the systems of said AIDS afflicted humans.

SUMMARY OF THE INVENTION

This invention relates to a molecular scale device not only capable of
destroying the AIDS virus, but of purging the human bloodstream of
pathogens and restoring immunity to AIDS patients of the candidiasis
and wasting syndrome categories. Said molecular device consists of a
single crystal of tetrasilver tetroxide (Ag.sub.4 O.sub.4). The
crystal lattice of this molecule has a unique structure since it is a
diamagnetic semiconducting crystal containing two mono and two
trivalent silver ions, which in effect are capable of "firing"
electrons under certain conditions which will destroy AIDS viruses,
other pathogens and immune suppressing moieties (ISM), not only
through the electrocution mode, but also by a binding process which
occurs simultaneously with electron firing, namely, binding and
chelation of divalent silver, i.e., the resulting product of the
electron transfer redox that occur when the monovalent silver ions are
oxidized and the trivalent ions are reduced in the crystal. The
binding/chelation effect occurs at active sites of the AIDS virus,
pathogens and ISM. Because of the extremely minute size of a single
molecule of this crystal, several million of these devices may be
employed in concert to destroy a virus colony to purge a life support
system of ISM and pathogens with the consumption of only parts per
trillion of the crystal devices. Thus an optimum of 40 PPM of the
devices by weight of human blood was found to be sufficient to
completely obliterate AIDS. This concentration is slightly over double
of the optimum concentration recommended in applicant's aforementioned
U.S. patent for the destruction of the human AIDS virus in vitro.
Other details concerning the structure of the crystal and its
mechanism against pathogens, the AIDS virus and ISM would analogously
hold here, and have already been further elucidated in said patent.

The actual destruction of pathogens, ISM and the AIDS virus is
effectuated by injection of a suspension of these devices in distilled
or deionized water with a non-reacting electrolyte directly, i.e.
intravenously, into the bloodstream. A single injection is all that is
required under these conditions. Accordingly, humans injected in this
manner, upon being inspected after three weeks or more had elapsed and
compared with similar humans that had been given placebos, were
completely cured of AIDS. The control group still manifested AIDS.
Accordingly, the tetrasilver tetroxide device performed in concert
with and in full conformity with the ultimate objects of this
invention. Furthermore, three out of four wasting syndrome terminal
patients and four out of the five candidiasis terminal patients were
still alive in 1995 after a year and a half had elapsed from their
initial injection. By that time all the AIDS patients had been
released from the clinic and allowed to return home.

Other objects and features of the present invention shall become
apparent to those skilled in the art when the present invention is
considered in view of the accompanying examples. It should, of course,
be recognized that the accompanying examples illustrate preferred
embodiments of the present invention and are not intended as a means
of defining the limits and scope of the present invention.

EXAMPLE 1

Five patients afflicted with AIDS of the candidiasis etiological
category were segregated for Tetrasil treatment. The rationale for
selecting them was based on facts presented in an article by Peter H.
Duesberg and Brian J. Ellison entitled "Is The AIDS Virus A Science
Fiction?" (Policy Review, Summer 1990 pp. 40-51). Only the factual
presentations of the article were utilized and the hypothesis of the
authors was ignored. The facts presented in the article related to the
method of selecting AIDS patients based on the five aforementioned
etiological subgroups targeted by the CDC, and the evidence presented,
that there is AIDS without HIV as well as with it so that an anti-
viral agent in most instances will not necessarily restore the
immunity system.

Evaluations with Tetrasil were conducted on AIDS patients at Lucha
Contra el Sida, Comayaguela, Honduras. The patients two weeks prior to
inoculation were removed from their AZT, AIDS therapy. Tetrasil was
administered at approximately 40 PPM of blood volume per patient as a
suspension in a proprietary buffer solution (pH=6.5), supplied by
Holipharm Corporation.

The results of evaluations with candidiasis are tabulated in Table I
under its disease category. All patients evaluated were terminal.
Some, however, were in moderate (m) condition and others in poor (p)
as designated in the Table. The I and F designations refer to initial
and final values as shown. WBC indicates white cell blood count. The H
column, following CD 8, indicates whether hepatomegaly occurred. This
was an unfortunate consequence of the treatment which resulted in
enlarged livers in all patients except the second one. Despite
hepatomegaly, there was no interference with liver function.

The onset of hepatomegaly was not spontaneous and varied from patient
to patient, being in the range of 4-16 days.

It should also be noted that shortly after injection of Tetrasil there
were indications of fever (symbolized by T in the Ag.sub.4 O.sub.4
column), sometimes accompanied by fatigue (F). The body temperature
was invariably 38.5.degree. C. (101.3.degree. F.). This was indicative
of restoration of the immune response of the body, since normally the
body will destroy pathogens when the immune system is functional by
raising the temperature. The patient who died; first responded
favorably to Diflucan, which previously gave no response. He was cured
of his candidiasis, but unfortunately succumbed to his previous body
damage. All the other candidiasis syndrome people who previously did
not respond to the indicated medications subsequently responded after
the Tetrasil treatment. Further evidence of the recovery of the AIDS
patients manifested itself 30 days after the initial injection when
white blood cell counts were taken. They are shown in Table I under
the WBC column, which gives the initial and final WBC. All candidiasis
patients showed a dramatic increase in their white blood cell counts,
indicative of the restoration of their immunity systems.

EXAMPLE 2

The above protocol of Example 1 was repeated with AIDS patients
exhibiting wasting syndrome. The results of their treatment are
tabulated in Table I under the disease category of said syndrome. It
should be noted that two of the four wasting syndrome patients showed
improved white blood counts. The female patient, whose condition
improved from poor and terminal to be among the living, showed a
decrease in the WBC. However, she showed an increase in body
temperature which was indicative of immune response. The test results
indicate that one cannot rely on a single factor to indicate the
demise of AIDS. The usual HIV marker CD 4 initial and final are
irrelevant. ISM suppression appears to be more critical than the
destruction of HIV. AIDS was suppressed, any permanent damage that had
been done to the patients in the course of their succumbing to AIDS
was not obviously cured or corrected by said crystal device treatment,
rather said injury persisted and the patient was improved with respect
to AIDS but still suffered from said permanent injury or impairment
previously inflicted.

TABLE I
__________________________________________________________________________
Response of AIDS Patients to Single 40 PPM Ag.sub.4 O.sub.4
Inoculation Date Weight DISEASE PATIENT Inoc. WBC CD 4 DEATH Lbs.
Group Sex Age Medictn 1994 I F I F CD 8 H 1944 I F Ag.sub.4 O.sub.4
__________________________________________________________________________
Candidiasis M p 28 Diflucan 5/5 1,200 4,200 41 -- 221 + 6/11 82 76 T F
m 33 " 5/5 6,000 6,700 554 872 394 - 98 98 T F m 33 Ketaconzl 5/27
2,600 3,850 248 181 951 + 123 123 T M p 62 " 6/2 3,300 3,700 89 237 59
+ 105 92 F F m 31 Pentamidn 6/2 2,400 3,050 9 181 65 + 121 118 Pain
Wasting M m 27 5/27 3,600 4,600 39 14 709 + 119 120 T Syndrome M m 28
5/27 2,750 -- 10 -- 60 + 7/19 121 119 T, F F p 43 5/27 3,600 2,700 68
246 248 + 101 98 T, F M m 19 5/10 3,850 5,400 137 36 48 + 103 106 T, F
__________________________________________________________________________

As this invention may be embodied in several forms without departing
from the spirit or essential characteristics thereof, the present
embodiments are therefore illustrative and not restrictive, since the
scope of the invention is defined by the appended claims rather than
by the description preceding them, and all changes that fall within
the metes and bounds of the claims or that form their functional as
well as conjointly cooperative equivalents, are therefore intended to
be embraced by these claims.




United States Patent 5,676,977
Antelman October 14, 1997
Method of curing AIDS with tetrasilver tetroxide molecular crystal
devices

Abstract

The diamagnetic semiconducting molecular crystal tetrasilver tetroxide
(Ag.sub.4 O.sub.4) is utilized for destroying the AIDS virus,
destroying AIDS synergistic pathogens and immunity suppressing
moieties (ISM) in humans. A single intravenous injection of the
devices is all that is required for efficacy at levels of about 40 PPM
of human blood. The device molecular crystal contains two mono and two
trivalent silver ions capable of "firing" electrons capable of
electrocuting the AIDS virus, pathogens and ISM. When administered
into the bloodstream, the device electrons will be triggered by
pathogens, a proliferating virus and ISM, and when fired will
simultaneously trigger a redox chelation mechanism resulting in
divalent silver moieties which chelate and bind active sites of the
entities destroying them. The devices are completely non-toxic.
However, they put stress on the liver causing hepatomegaly, but there
is no loss of liver function.
Inventors: Antelman; Marvin S. (Rehovot, IL)
Assignee: Antelman Technologies Ltd. (Providence, RI)
Appl. No.: 08/658,955
Filed: May 31, 1996
Related U.S. Patent Documents

Application Number Filing Date
Patent Number Issue Date
310859 Sep., 1994

Current U.S. Class: 424/618 ; 514/495
Current International Class: A61K 33/38 (20060101); A61K 033/38 ()
Field of Search: 424/618 514/495
References Cited [Referenced By]
U.S. Patent Documents

4415565 November 1983 Wysor
4915955 April 1990 Gomori
4952411 August 1990 Fox, Jr. et al.
5073382 December 1991 Antelman
5078902 January 1992 Antelman
5089275 February 1992 Antelman
5211855 May 1993 Antelman
5223149 June 1993 Antelman
5336499 August 1994 Antelman
5571520 November 1996 Antelman

Other References

"Is The AIDS Virus A Science Fiction?" by Peter H. Duesberg and Bryan
J. Ellison, Policy Review, Summer 1990, pp. 40-51..

Primary Examiner: Hulina; Amy
Attorney, Agent or Firm: Salter & Michaelson
Parent Case Text


This application is a continuation-in-part of patent application Ser.
No. 08/310,859 filed Sep. 22, 1994,
now abandoned.
Claims

What is claimed is:

1. A method of treating AIDS-afflicted humans comprising injecting a
multitude of tetrasilver tetroxide molecular crystals into the
bloodstream of the human subject.

2. A method for increasing white blood cell counts in AIDS-afflicted
humans comprising injecting a multitude of tetrasilver tetroxide
molecular crystals into the bloodstream of the human subject.

3. Methods of treating AIDS-affilicted humans according to claims 1-2
where the concentration of said molecular crystals is approximately 40
PPM of the total blood weight of the human subject.
Description


BACKGROUND OF THE INVENTION

The present invention relates to the employment of molecular crystals
as anti-AIDS devices, but more particularly to the molecular crystal
semiconductor tetrasilver tetroxide Ag.sub.4 O.sub.4 which has two
monovalent and two trivalent silver ions per molecule, and which
through this structural configuration enables intermolecular electron
transfer capable of killing viruses and binding them to the resulting
silver entity so that a single intravenous injection will completely
obliterate acquired immune deficiency syndrome (AIDS) in humans.
Furthermore, said devices are capable of killing pathogens and purging
the bloodstream of immune suppressing moieties (ISM) whether or not
created by the AIDS virus (HIV); so as to restore the immune system.

The present invention is based on concepts previously elucidated in
applicant's U.S. Pat. No. 5,336,499 which discloses the destruction
and inhibition of bacteria, algae and the AIDS virus in nutrient life
supporting systems by using said silver oxide devices. Example 3 of
said patent discloses that 18 PPM of said crystal devices could
totally suppress the AIDS virus (page 6, line 5). Subsequent to the
filing of the aforementioned patent, further testing revealed complete
100% destruction of the AIDS virus in vitro at 20 PPM, and the fact
that said devices were harmless when ingested and inhaled, being non-
toxic.

Encouraged by these evaluations and successes, applicant obtained
permission to evaluate the crystals in vitro against murine acquired
immune deficiency syndrome (MAIDS). Only one facility in the State of
Israel is licensed for these evaluations, namely, the Kaplan Hospital
in Rehovot, Israel, which is affiliated with the Hebrew University-
Hadassah Medical School where said evaluations were done.

The initial evaluations entailed experimenting with various silver
moieties cited in applicant's aforementioned patent, concentrations,
non-reactive buffers and modes of administration. After about 18
months of judicious efforts and initial failures, success was finally
achieved in destroying the MAIDS virus in C57BL mice with a single
intravenous injection. The results of this test program comprise
Example 5 of U.S. Pat. No. 5,336,499. After success with mice, the
inventor was able to test the efficacy of said devices on two select
etiological groups of terminal AIDS patients in a clinic in
Tegucigalpa, Honduras, Central America.

The AIDS patients comprised the etiological subgroups, Candidiasis and
Wasting Syndrome. Current indicator diseases for diagnosing AIDS which
have been expanded by the CDC, fall into the following five major
categories with the approximate percent distribution among AIDS
patients:

______________________________________ 1. P. carinii pneumonia 51% 2.
Wasting syndrome 19% 3. Candidiasis 13% 4. Kaposi's sarcoma 11% 5.
Dementia 6% ______________________________________

This invention concerns itself with the treatment and cure of
candidiasis and wasting syndrome AIDS patients with Tetrasil*. These
two groups account for approximately one third of AIDS cases.

Stedman's Medical Dictionary (Williams & Wilken's 26th Ed., 1995)
defines wasting syndrome "as a condition of 10% weight loss in
conjunction with diarrhea or fever . . . Associated with AIDS (p.
1744)."

OBJECTS OF THE INVENTION

The main object of the invention is to provide for a molecular scale
device of a single tetrasilver tetroxide crystalline molecule capable
of restoring the immunity of AIDS afflicted humans of the two AIDS
etiological subgroups, candidiasis and wasting syndrome.

Another object of the invention is to provide for immunity restoration
in said AIDS afflicted humans through a single injection.

Another object of this invention is to destroy ISM in humans
manifesting AIDS diseases of said AIDS etiological subgroups
irrespective as to whether said ISM was HIV induced, since it is known
that humans may manifest AIDS and still be HIV negative, and thus
restore the immune system in said humans.

Another object of this invention is to destroy the AIDS virus when
present in the systems of said AIDS afflicted humans.

SUMMARY OF THE INVENTION

This invention relates to a molecular scale device not only capable of
destroying the AIDS virus, but of purging the human bloodstream of
pathogens and restoring immunity to AIDS patients of the candidiasis
and wasting syndrome categories. Said molecular device consists of a
single crystal of tetrasilver tetroxide (Ag.sub.4 O.sub.4). The
crystal lattice of this molecule has a unique structure since it is a
diamagnetic semiconducting crystal containing two mono and two
trivalent silver ions, which in effect are capable of "firing"
electrons under certain conditions which will destroy AIDS viruses,
other pathogens and immune suppressing moieties (ISM), not only
through the electrocution mode, but also by a binding process which
occurs simultaneously with electron firing, namely, binding and
chelation of divalent silver, i.e., the resulting product of the
electron transfer redox that occur when the monovalent silver ions are
oxidized and the trivalent ions are reduced in the crystal. The
binding/chelation effect occurs at active sites of the AIDS virus,
pathogens and ISM. Because of the extremely minute size of a single
molecule of this crystal, several million of these devices may be
employed in concert to destroy a virus colony to purge a life support
system of ISM and pathogens with the consumption of only parts per
trillion of the crystal devices. Thus an optimum of 40 PPM of the
devices by weight of human blood was found to be sufficient to
completely obliterate AIDS. This concentration is slightly over double
of the optimum concentration recommended in applicant's aforementioned
U.S. patent for the destruction of the human AIDS virus in vitro.
Other details concerning the structure of the crystal and its
mechanism against pathogens, the AIDS virus and ISM would analogously
hold here, and have already been further elucidated in said patent.

The actual destruction of pathogens, ISM and the AIDS virus is
effectuated by injection of a suspension of these devices in distilled
or deionized water with a non-reacting electrolyte directly, i.e.
intravenously, into the bloodstream. A single injection is all that is
required under these conditions. Accordingly, humans injected in this
manner, upon being inspected after three weeks or more had elapsed and
compared with similar humans that had been given placebos, were
completely cured of AIDS. The control group still manifested AIDS.
Accordingly, the tetrasilver tetroxide device performed in concert
with and in full conformity with the ultimate objects of this
invention. Furthermore, three out of four wasting syndrome terminal
patients and four out of the five candidiasis terminal patients were
still alive in 1995 after a year and a half had elapsed from their
initial injection. By that time all the AIDS patients had been
released from the clinic and allowed to return home.

Other objects and features of the present invention shall become
apparent to those skilled in the art when the present invention is
considered in view of the accompanying examples. It should, of course,
be recognized that the accompanying examples illustrate preferred
embodiments of the present invention and are not intended as a means
of defining the limits and scope of the present invention.

EXAMPLE 1

Five patients afflicted with AIDS of the candidiasis etiological
category were segregated for Tetrasil treatment. The rationale for
selecting them was based on facts presented in an article by Peter H.
Duesberg and Brian J. Ellison entitled "Is The AIDS Virus A Science
Fiction?" (Policy Review, Summer 1990 pp. 40-51). Only the factual
presentations of the article were utilized and the hypothesis of the
authors was ignored. The facts presented in the article related to the
method of selecting AIDS patients based on the five aforementioned
etiological subgroups targeted by the CDC, and the evidence presented,
that there is AIDS without HIV as well as with it so that an anti-
viral agent in most instances will not necessarily restore the
immunity system.

Evaluations with Tetrasil were conducted on AIDS patients at Lucha
Contra el Sida, Comayaguela, Honduras. The patients two weeks prior to
inoculation were removed from their AZT, AIDS therapy. Tetrasil was
administered at approximately 40 PPM of blood volume per patient as a
suspension in a proprietary buffer solution (pH=6.5), supplied by
Holipharm Corporation.

The results of evaluations with candidiasis are tabulated in Table I
under its disease category. All patients evaluated were terminal.
Some, however, were in moderate (m) condition and others in poor (p)
as designated in the Table. The I and F designations refer to initial
and final values as shown. WBC indicates white cell blood count. The H
column, following CD 8, indicates whether hepatomegaly occurred. This
was an unfortunate consequence of the treatment which resulted in
enlarged livers in all patients except the second one. Despite
hepatomegaly, there was no interference with liver function.

The onset of hepatomegaly was not spontaneous and varied from patient
to patient, being in the range of 4-16 days.

It should also be noted that shortly after injection of Tetrasil there
were indications of fever (symbolized by T in the Ag.sub.4 O.sub.4
column), sometimes accompanied by fatigue (F). The body temperature
was invariably 38.5.degree. C. (101.3.degree. F.). This was indicative
of restoration of the immune response of the body, since normally the
body will destroy pathogens when the immune system is functional by
raising the temperature. The patient who died; first responded
favorably to Diflucan, which previously gave no response. He was cured
of his candidiasis, but unfortunately succumbed to his previous body
damage. All the other candidiasis syndrome people who previously did
not respond to the indicated medications subsequently responded after
the Tetrasil treatment. Further evidence of the recovery of the AIDS
patients manifested itself 30 days after the initial injection when
white blood cell counts were taken. They are shown in Table I under
the WBC column, which gives the initial and final WBC. All candidiasis
patients showed a dramatic increase in their white blood cell counts,
indicative of the restoration of their immunity systems.

EXAMPLE 2

The above protocol of Example 1 was repeated with AIDS patients
exhibiting wasting syndrome. The results of their treatment are
tabulated in Table I under the disease category of said syndrome. It
should be noted that two of the four wasting syndrome patients showed
improved white blood counts. The female patient, whose condition
improved from poor and terminal to be among the living, showed a
decrease in the WBC. However, she showed an increase in body
temperature which was indicative of immune response. The test results
indicate that one cannot rely on a single factor to indicate the
demise of AIDS. The usual HIV marker CD 4 initial and final are
irrelevant. ISM suppression appears to be more critical than the
destruction of HIV. AIDS was suppressed, any permanent damage that had
been done to the patients in the course of their succumbing to AIDS
was not obviously cured or corrected by said crystal device treatment,
rather said injury persisted and the patient was improved with respect
to AIDS but still suffered from said permanent injury or impairment
previously inflicted.

TABLE I
__________________________________________________________________________
Response of AIDS Patients to Single 40 PPM Ag.sub.4 O.sub.4
Inoculation Date Weight DISEASE PATIENT Inoc. WBC CD 4 DEATH Lbs.
Group Sex Age Medictn 1994 I F I F CD 8 H 1944 I F Ag.sub.4 O.sub.4
__________________________________________________________________________
Candidiasis M p 28 Diflucan 5/5 1,200 4,200 41 -- 221 + 6/11 82 76 T F
m 33 " 5/5 6,000 6,700 554 872 394 - 98 98 T F m 33 Ketaconzl 5/27
2,600 3,850 248 181 951 + 123 123 T M p 62 " 6/2 3,300 3,700 89 237 59
+ 105 92 F F m 31 Pentamidn 6/2 2,400 3,050 9 181 65 + 121 118 Pain
Wasting M m 27 5/27 3,600 4,600 39 14 709 + 119 120 T Syndrome M m 28
5/27 2,750 -- 10 -- 60 + 7/19 121 119 T, F F p 43 5/27 3,600 2,700 68
246 248 + 101 98 T, F M m 19 5/10 3,850 5,400 137 36 48 + 103 106 T, F
__________________________________________________________________________

As this invention may be embodied in several forms without departing
from the spirit or essential characteristics thereof, the present
embodiments are therefore illustrative and not restrictive, since the
scope of the invention is defined by the appended claims rather than
by the description preceding them, and all changes that fall within
the metes and bounds of the claims or that form their functional as
well as conjointly cooperative equivalents, are therefore intended to
be embraced by these claims.




United States Patent 7,687,076
Djokic March 30, 2010
Deposition products, composite materials and processes for the
production thereof

Abstract

A method for producing a deposition product includes providing a
deposition solution having a pH below 7 and comprising an aqueous
solution of a silver salt comprised of silver ions and an oxidizing
agent. The silver salt is comprised of silver nitrate and has a
concentration in the deposition solution between about 1 and 20 grams
per liter. The oxidizing agent is comprised of a persulfate and has a
concentration in the deposition solution between about 1 and 50 grams
per liter. The method further includes producing the deposition
product by facilitating a chemical reaction between the silver ions
and the oxidizing agent while maintaining the deposition solution at a
temperature of between about 2 and 40 degrees Celsius. The deposition
product consists essentially of at least one oxidized silver species,
and is comprised of a compound having the formula Ag.sub.7O.sub.8X,
where X is an anion.
Inventors: Djokic; Stojan (Edmonton, CA)
Assignee: Exciton Technologies Inc. (CA)
Appl. No.: 11/934,459
Filed: November 2, 2007
Related U.S. Patent Documents

Application Number Filing Date Patent Number
Issue Date
10830574 Apr., 2004 7300673
Foreign Application Priority Data

May 16, 2003 [CA] 2428922
Mar 10, 2004 [CA] 2460585

Current U.S. Class: 424/618 ; 424/443; 424/445; 424/613; 424/703;
514/495; 514/709
Current International Class: A61K 33/38 (20060101); A61K 31/10
(20060101); A61K 33/40 (20060101); A61K 9/70 (20060101); A61L 15/00
(20060101); A61K 33/04 (20060101); A61K 31/28 (20060101); A01N 39/00
(20060101); A01N 41/10 (20060101); A01N 55/02 (20060101); A01N 59/02
(20060101); A01N 59/16 (20060101)
References Cited [Referenced By]
U.S. Patent Documents

4297249 October 1981 Przybyla et al.
4728323 March 1988 Matson
5098582 March 1992 Antelman
5211855 May 1993 Antelman
5372847 December 1994 Silver et al.
5413788 May 1995 Edwards et al.
5474797 December 1995 Sioshansi et al.
5676977 October 1997 Antelman
5681575 October 1997 Burrell et al.
5814094 September 1998 Becker et al.
5928174 July 1999 Gibbins
5985308 November 1999 Burrell et al.
6017553 January 2000 Burrell et al.
6080490 June 2000 Burrell et al.
6087549 July 2000 Flick
6224983 May 2001 Sodervall et al.
6238686 May 2001 Burrell et al.
6267782 July 2001 Ogle et al.
6306341 October 2001 Yokota et al.
6333093 December 2001 Burrell et al.
6355858 March 2002 Gibbins
6379712 April 2002 Yan et al.
6426195 July 2002 Zhong et al.
6436420 August 2002 Antelman
6444109 September 2002 Redline et al.
2001/0009831 July 2001 Schink et al.
2001/0024662 September 2001 Yang
2002/0051824 May 2002 Burrell et al.
2002/0127282 September 2002 Antelman
Foreign Patent Documents

0875146 Apr., 1998 CA
2102433 Mar., 2000 CA
1149389 May., 1997 CN
431656 Jul., 1935 GB
WO2004037187 May., 2004 WO

Other References

NR. Thompson, Silver, in Comprehensive Inorganic Chemistry, v.IIID,
J.C. Bailer et al, Editors, Pergamon Press (1973) 79-82. cited by
other .
K.J. Bundy et al, "An Investigation of the Bacteriostatic Properties
of pure Metals", Journal of Biomedical Materials Research, v. 14
(1980) 653-662. cited by other .
D. Acel, "Uber die oligodynamishe Wirkung der Metalle," Z. Biochem,
112 (1920) 23-26 with English Abstract. cited by other .
J. Gibbard, "Public Health Aspects of the Treatment of Water and
Beverages with Silver", Journal of American Public Health, v. 27
(1937) 112-11. cited by other .
S.S. Djokic and R.E. Burrell, "Behavior of Silver in Physiological
Solutions", Journal of Electrochemical Society, v. 145 (5) (1998)
1426-1430. cited by other .
S.S. Djokic et al, "An Electrochemical Analysis of Thin Silver
Produced by Reactive Sputtering", J. of Electrochemical Society v.
148(3) (2001) C191-C196. cited by other .
C.L. Fox, "Topical Therapy and the Development of Sulfadiazine",
Surgery, Gynecology & Obstetrics, 157 (1983) 82-88. cited by other .
M.C. Fung, D.L. Bowen, "Silver Products for Medical Indications: Risk-
Benefit Assessment", Clinical Toxicology, v. 34 (1) (1996) 119-126.
cited by other .
H.W. Margaf, T.H. Covey, "A Trial of Silver-Zinc-Allantoinate in the
Treatment of Leg Ulcers", Arch. Surg. v. 112 (1977) 699-704. cited by
other .
S.I. Zhdanov, "Sulfur, Selenium, Tellurium and Polonium", in Standard
Potentials in Aqueous Solutions, A.J. Bard et al, Editors, Marcel
Dekker Inc., New York (1985) 93, 5pp. cited by other .
M.S. Skanavi-Grigoreva, I.L. Shimanovich, Zh. Obsh., Khim., 24, (1954)
1490-1495 with English abstract. cited by other .
"Electrocrystallization" article from website: www.mpi-stuttgart.mpg.de/jansen/p110,
2pp. cited by other .
Leibecki, Harold F., "Argentic Oxysalt Electrodes", NASA Technical
Note NASA TN D-3208, Washington, D.C., Jan. 1966. cited by other .
McMillan, J.A., "Higher Oxidation of States of Silver", Chem. Rev.
(Washington D.C.) 62, 1962, pp. 65-80. cited by other .
Discussion Section, Journal of The Electrochemical Society, vol. 106,
No. 12, Dec. 1959, pp. 1072-1084. cited by other.

Primary Examiner: Arnold; Ernst V
Attorney, Agent or Firm: Kuharchuk; Terrence N. Rodman & Rodman
Claims


The embodiments of the invention in which an exclusive property or
privilege is claimed are defined as follows:

1. A method for producing a deposition product, the method comprising
the following steps: (a) providing a deposition solution comprising an
amount of an aqueous solution of a silver salt comprised of an amount
of silver ions and an amount of an oxidizing agent, wherein the silver
salt is comprised of silver nitrate, wherein the oxidizing agent is
comprised of a persulfate, wherein a concentration of the silver salt
in the deposition solution is between about 1 gram per liter and about
20 grams per liter, wherein a concentration of the oxidizing agent in
the deposition solution is between about 1 gram per liter and about 50
grams per liter, and wherein the deposition solution has a pH below 7;
and (b) producing the deposition product by facilitating a chemical
reaction in the deposition solution between the silver ions and the
oxidizing agent while maintaining the deposition solution at a
temperature of between about 2 degrees Celsius and about 40 degrees
Celsius, wherein the deposition product consists essentially of at
least one oxidized silver species, and wherein the deposition product
is comprised of a compound having the formula Ag.sub.7O.sub.8X, where
X is an anion.

2. The method as claimed in claim 1 wherein the persulfate is selected
from the group of persulfates consisting of potassium persulfate,
sodium persulfate, ammonium persulfate and mixtures thereof.

3. The method as claimed in claim 2 wherein the persulfate is
comprised of potassium persulfate.

4. The method as claimed in claim 1 wherein the amount of the
oxidizing agent is selected to be a stoichiometrically appropriate
amount relative to the amount of the silver ions.

5. The method as claimed in claim 2, further comprising the step of
adding an amount of a source of anions to the deposition solution for
combining with the silver ions in order to produce the deposition
product.

6. The method as claimed in claim 5 wherein the source of anions is
comprised of at least one acid.

7. The method as claimed in claim 6 wherein the acid is selected from
the group of acids consisting of carbonic acid, nitric acid,
perchloric acid, sulfuric acid, acetic acid, fluoroboric acid, citric
acid, acetylsalicylic acid and mixtures thereof.

8. The method as claimed in claim 5 wherein the amount of the source
of anions is selected to be a stoichiometrically appropriate amount
relative to the amount of the silver ions.

9. The method as claimed in claim 2 wherein the deposition product
producing step is comprised of maintaining the deposition solution at
a temperature of between about 10 degrees Celsius and about 25 degrees
Celsius.

10. The method as claimed in claim 2 wherein the deposition product
producing step is comprised of agitating the deposition solution
during at least a portion of the deposition product producing step.

11. The method as claimed in claim 1 wherein the deposition product is
further comprised of Ag.sub.2SO.sub.4.

12. The method as claimed in claim 1 wherein the deposition product is
further comprised of at least one silver oxide selected from the group
of silver oxides consisting of monovalent silver oxide, bivalent
silver oxide, trivalent silver oxide and mixtures thereof.

13. The method as claimed in claim 1 wherein X is derived from an
acid.

14. The method as claimed in claim 1 wherein the deposition product is
comprised of a plurality of valent states of silver.

15. The method as claimed in claim 1 wherein X is selected from the
group of anions consisting of HCO.sub.3.sup.-, CO.sub.3.sup.2-, NO.sub.
3.sup.-, ClO.sub.4.sup.-, SO.sub.4.sup.2-, F.sup.-, and mixtures
thereof.

16. The method as claimed in claim 15 wherein X is comprised of NO.sub.
3.sup.-.
Description


FIELD OF INVENTION

Deposition products, composite materials including deposition
products, and methods for producing the deposition products and the
composite materials.

BACKGROUND ART

The germicidal properties of silver, even not known as such, have been
utilized since the early Mediterranean cultures. It has been known
since 1000 BC and possibly before that water kept in silver vessels
and then exposed to light and filtered could be rendered potable.
Other forms of silver have been used throughout centuries for various
applications, such as coatings for prevention of beverages from
spoilage or silver plates and foils in the surgical treatments of
wounds and broken bones.

The lethal effects of metals towards bacteria and lower life forms
were first scientifically described by von Nageli in the late
nineteenth century, and this phenomenon has been defined as an
"oligodynamic effect" (N. R. Thompson, Silver, in Comprehensive
Inorganic Chemistry, Vol. III D, J. C. Bailer, H. J. Emeleus, R.
Nyholm and A. F. Trutman-Dickenson, Editors, Pergamon Press, Oxford
(1973)). The term oligodynamic effect is typically restricted to
describing solutions in which the metal concentration is several
orders of magnitude lower than that which would be lethal to higher
organisms.

The investigation of the bacteriostatic properties of pure metals such
as Fe, Mo, Cu, V, Sn, W, Au, Al, Ta, Nb, Ti, Zr, Ni, Co, Ag and Cr,
has proved that Co was the only element which was inhibitory for the
bacterial growth under anaerobic conditions (K. J. Bundy, M. F. Butler
and R. F. Hochman, "An Investigation of the Bacteriostatic Properties
of Pure Metals", Journal of Biomedical Materials Research, Vol. 14
(1980) 653-663). Under aerobic conditions both Cu and Co consistently
display inhibitory effects. Some antimicrobial effects have been seen
for Ni, Fe and V. However, other metals such as Mo, W, Al, Nb, Zr, Cr
and most importantly for the present invention Ag and Sn never showed
any tendency to inhibit the growth of Streptococcus mutans.

In the case of silver metal, it was in 1920, when Acel who was the
first to attribute the antimicrobial properties of silver to the
liberation of Ag.sup.+ ions from the material (D. Acel, "Uber die
oligodynamische Wirkung der Metalle", Z. Biochem., 112 (1920) 23).

Gibbard reported in 1937 that pure metallic silver has no
antimicrobial activity (J. Gibbard, "Public Health Aspects of the
Treatment of Water and Beverages with Silver", Journal of American
Public Health, Vol. 27 (1937) 112-119). His experiments showed that if
silver is cleaned mechanically with an abrasive cloth or paper it
becomes inactive. Similarly, if molten silver is allowed to cool in a
reduction atmosphere (e.g. hydrogen), no antimicrobial activity is
found. When cooling of molten silver is carried out in air, and
formation of surface oxide occurred, an antimicrobial activity may be
observed. Similar results were found when silver metal was treated
with nitric acid in an air atmosphere (dissolution and formation of an
oxide layer). Based on Gibbard's results, pure silver was devoid of
activity, but surface oxidized silver was active. Silver oxide, silver
nitrate and silver chloride were always active. Also, Gibbard observed
that the antimicrobial properties of silver and its compounds were
reduced in the presence of proteins or glucose.

Djoki investigated the behavior of silver films, e.g. physical vapor
deposited, electrodeposited, electroless deposited and metallurgical
in physiological saline solutions (S. S. Djoki and R. E. Burrell,
"Behavior of Silver in Physiological Solutions", Journal of the
Electrochemical Society, Vol. 145 (5) (1998) 1426-1430). Djoki found
that an essential factor leading to an antimicrobial activity of
metallic silver is a presence of Ag oxide(s) at the surface of this
material. It was demonstrated that only silver films containing silver
oxides (most likely Ag.sub.2O) showed an antimicrobial activity. The
behavior was attributed to the dissolution of Ag.sub.2O from the
"silver" material and formation of Ag.sup.+ or other complexed ions
which become antimicrobially active. There was no evidence that pure
metallic silver, no matter which way it was produced i.e., physical
vapor deposited, electrodeposited or electroless deposited could be
dissolved in physiological media, or that these materials would
exhibit antimicrobial activity.

It should be noted that when the physical vapor deposition of silver
was carried out in an atmosphere containing oxygen the resulted
product, as found by the XRD analysis contained silver oxide.
Consequently, these samples exhibited antimicrobial activity.
Conversely, when the physical vapor deposition was carried out from an
argon atmosphere (no presence of oxygen) pure metallic,
nanocrystalline silver film was deposited as confirmed by the XRD
analysis. However, these films did not dissolve in physiological
saline solutions, nor they exhibited antimicrobial activity at all.

For an in depth understanding of structural properties of silver films
produced by reactive sputtering, see Djoki et al. (S. S. Djoki , R. E.
Burrell, N. Le and D. J. Field, "An Electrochemical Analysis of Thin
Silver Produced by Reactive Sputtering", Journal of the
Electrochemical Society, Vol. 148 (3) (2001) C191-C196.). To prove the
concept that only oxidized silver species are responsible for the
antimicrobial activity, Djoki further oxidized pure metallic silver
samples (i.e. those produced by the electrodeposition, electroless
deposition, physical vapor deposition in an argon atmosphere or
metallurgically). The oxidation of these samples was carried out
electrochemically in 1 M KOH solutions, using a process very well
established in the art. The electrochemically oxidized silver samples
were tested for the antimicrobial activity against Pseudomonas
aeruginosa. Clear evidence was found that the electrochemically
oxidized silver samples exhibited antimicrobial activity.

The above referenced work shows that only oxidized silver species, but
not elemental silver will affect antimicrobial activity. The findings
to date show that the "nanocrystalline" or "macrocrystalline"
elemental silver does not have antimicrobial activity at all.
Elemental silver, either nanocrystalline or "macrocrystalline" may
exhibit some antimicrobial activity only if oxidized silver species
are present at these surfaces or within the silver metal. Only the
formation of silver oxide(s), carbonates or other silver salts (except
silver sulfide, due to its extremely low solubility) at the surface or
within the material, which may be influenced by an exposure of
elemental silver to various bases, acids or due to atmospheric
corrosion may lead to an antimicrobial activity of this material.

The use of silver on chronic wounds dates back in the 17.sup.th and
18.sup.th centuries. In the early 19.sup.th century, silver nitrate
began to be used on burns and in opthalmology. Concentrations of the
solution ranged from 0.20 to 2.5 wt. % with the weaker solutions being
reserved for children. Silver has been found to be active against a
wide range of bacterial, fungal and viral pathogens. Topical treatment
of acute and chronic wounds is a preferred and selective approach to
the prevention of infection and healing. In order to achieve these
requirements products that are used in the prevention of infections
must have certain physical and chemical properties.

When used for topical dressings, silver compounds must have relatively
low solubility. This is usually achieved by choosing compounds with a
relatively low solubility products (e.g. AgCl, Ag.sub.2SO.sub.4,
Ag.sub.2CO.sub.3, Ag.sub.3PO.sub.4, Ag-oxides). Kinetics of
dissolution of these compounds in neutral aqueous solutions is quite
slow. This property is very convenient for two reasons. First, a
sustained release of silver ions from the silver compounds is more
likely to provide a prolonged antimicrobial activity. Second, low
amounts of the silver ions released into wound exudates may not give
rise to transient high tissue blood and urine levels, thus avoiding
systemic toxicity. The choice of a particular silver compound will
depend upon its reactivity with wound exudates. This reactivity should
preferably be minimized in order to achieve the desired effect of the
released silver ions (i.e., antimicrobial activity without systemic
toxicity).

Besides silver nitrate, one of the most widely used topical
antimicrobial materials is silver sulfadiazine (C. L. Fox, "Topical
Therapy and the development of Silver Sulfadiazine", Surgery,
Gynecology & Obstetrics, 157 (1) (1983) 82-88). This compound is
synthesized from silver nitrate and sodium sulfadiazine. Silver
sulfadiazine has been used in treatments of burns, leg ulcers and also
as a topical antimicrobial agent in the management of infected wounds.

Products such as silver protein (argyrols) or mild silver protein are
mixtures of silver nitrate, sodium hydroxide and gelatin. These
products are recommended for internal use and are promoted as
essential mineral supplements. Although there is no theoretical or
practical justification for their use, this class of compounds has
been recommended for the treatment of diverse diseases such as cancer,
diabetes, AIDS and herpes (M. C. Fung, D. L. Bowen, "Silver Products
for Medical Indications: Risk--Benefit Assessment", Clinical
Toxicology, Vol. 34 (1) (1996) 119-126).

Silver-zinc-allantoinate has been formulated as a cream and represents
a combination of silver, zinc and allantoin (an agent that stimulates
debridement and tissue growth (H. W. Margaf, T. H. Covey, "A Trial of
Silver-Zinc-Allantoinate in the Treatment of Leg Ulcers", Arch. Surg.,
Vol. 112 (1977) 699-704). This composition exhibited promising effects
in preliminary studies.

In the past few decades several topical dressings containing silver
have been developed for wound care. Such materials include
Arglaes.TM., Silverlon.TM., Acticoat.TM., Actisorb.TM., and Silver
220.TM..

Antimicrobial coatings and methods of forming same are the subject of
U.S. Pat. No. 5,681,575 (Burrell et al) and U.S. Pat. No. 6,238,686
(Burrell et al). The coatings are formed by the physical vapour
deposition of biocompatible metal and the preferred biocompatible
metal is silver.

Burrell et al teach that atomic disorder may be created in metal
powders or foils by cold working and in metal coatings by depositing
by vapor deposition at low substrate temperatures and that such metal
coatings constitute a matrix containing atoms or molecules of a
different material. The presence of different atoms or molecules
results in atomic disorder in the metal matrix, for instance due to
different sized atoms. The different atoms or molecules may be one or
more second metals, metal alloys or metal compounds which are co- or
sequentially deposited with the first metal or metals to be released.
Alternatively, the different atoms or molecules may be adsorbed or
trapped from the working gas atmosphere during reactive vapor
deposition.

In U.S. Pat. No. 6,238,686 Burrell et al claim a modified material
comprising one or more metals in a form characterized by sufficient
atomic disorder such that the material, in contact with a solvent for
the material, releases atoms, ions, molecules or clusters containing
at least one metal at an enhanced rate relative to its normal ordered
crystalline structure. In U.S. Pat. No. 5,681,575 Burrell et al claim
a medical device which includes a coating of one or more anti-
microbial metals having a "sufficient atomic disorder".

It is unclear from either U.S. Pat. No. 5,681,575 or U.S. Pat. No.
6,238,686 what would constitute a material characterized by
"sufficient atomic disorder". In nature, most materials would exhibit
sufficient atomic disorder if the true atomic disorder described (by
drawings or mapping) in ordinary Chemistry or Physics handbooks were
insufficiently ordered (with a regular geometric structure or like).

In any event, the teachings of Burrell et al appear to connect "atomic
disorder" with an "enhanced rate" of release of "atoms, ions,
molecules or clusters". If the term "release" further relates to a
dissolution (as defined in textbooks of General Chemistry and
Physics), then this dissolution should lead to the liberation of ions
or molecules in solvent. When released in the solvent, these ions or
molecules are usually solvated i.e. surrounded by the molecules of the
solvent. It is very unlikely that atoms of a metal will be released
into a solution comprising of water such as in the wound environment.
If released into solution in its elemental state, metals would rather
represent a relatively larger particles comprising of more than one or
a few atoms.

As a result, the term "atom" as used in Burrell et al is not exactly
descriptive. It is not known yet scientifically whether atoms of
metals can be released into aqueous solutions at pH close to neutral
(e.g., pH range 6 to 8), except in the case of colloidal solutions
which are usually prepared by adequate chemical reactions in-situ.

U.S. Pat. No. 6,087,549 (Flick) discloses a multilayer laminate wound
dressing comprising a plurality of layers of a fibrous material, with
each layer comprising a unique ratio of metalized fibers to
nonmetalized fibers. In a preferred embodiment the wound dressing
consists of three layers and the metal is silver. The wound contact
layer has the highest ratio of metalized fibers to nonmetalized
fibers, the intermediate layer has a lower ratio of metalized fibers
to nonmetalized fibers, and the outer layer has the lowest ratio of
metalized fibers to nonmetalized fibers. The wound dressing described
by Flick is commercially available under the trade-mark Silverlon.TM..

U.S. Pat. No. 5,211,855 (Antelman), U.S. Pat. No. 5,676,977 (Antelman)
and U.S. Pat. No. 6,436,420 (Antelman) teach that tetrasilver
tetroxide (Ag.sub.4O.sub.4) containing two monovalent and two
trivalent silver ions exhibits bactericidal, fungicidal and algicidal
properties. As a result, "tetrasilver tetroxide" is suggested for use
for water treatment in U.S. Pat. No. 5,211,855 and for use in
destroying the AIDS virus in U.S. Pat. No. 5,676,977.

In U.S. Pat. No. 6,436,420, Antelman describes a method of deposition
or interstitial precipitation of tetrasilver tetroxide (Ag.sub.4O.sub.
4) crystals within the interstices of fibers, yarns and/or fabrics
forming such articles in order to produce fibrous textile articles
possessing enhanced antimicrobial properties. The interstitial
precipitation of Ag.sub.4O.sub.4 is achieved by immersion of the
article to be treated (e.g., fiber, yarn or fabric) in an aqueous
solution containing a water soluble silver salt, most preferably
silver nitrate. After uniformly wetting the article, the article is
removed into a second heated aqueous solution (having a temperature of
at least 85 degrees Celsius or more preferably at least 90 degrees
Celsius) containing strong alkali (most preferably NaOH) and a water
soluble oxidizing agent (most preferably potassium persulfate) for 30
seconds to 5 minutes to facilitate the precipitation of tetrasilver
tetroxide.

After the reaction is completed, the article is removed and washed.
The article treated in this way is described as exhibiting outstanding
antimicrobial resistance towards pathogens such as bacteria, viruses,
yeast and algae. The article is also described as being resistant to
ultraviolet light and as maintaining its antimicrobial properties
after a number of launderings.

SUMMARY OF INVENTION

The present invention is directed at deposition products, composite
materials and at methods for the production of deposition products and
composite materials. The deposition products are comprised of at least
one oxidized species of a metal.

The methods of the invention are based upon chemical deposition
principles and techniques. The methods of the invention may be carried
out under either acidic or alkaline conditions. The methods of the
invention may comprise the step of exposing ions of the metal to an
oxidizing agent to produce the deposition product. The methods of the
invention may involve the production of the deposition product itself
or the production of a composite material which comprises a substrate
and the deposition product.

The methods of the invention are particularly suited for producing a
composite material which is comprised of a substrate and a very thin
coating or deposition layer of the deposition product. This thin
coating or layer may be in the order of one or several atoms in
thickness, which facilitates the production of a composite material
which has a relatively high surface area to volume ratio. The coating
may also be deposited so that it does not completely cover the
substrate, thus leaving portions of the surface of the substrate
uncoated. Composite materials produced using the methods of the
invention may be useful for a variety of applications, including but
not limited to electronics, materials engineering and medical
applications.

The methods of the invention may be carried out at relatively low
temperatures. Preferably the methods of the invention are carried out
at temperatures of no greater than about 60 degrees Celsius. More
preferably the methods of the invention are carried out at room
temperature (i.e., between about 10 degrees Celsius and about 25
degrees Celsius).

The metal and the oxidizing agent are selected so that they are
compatible with the production of the desired deposition product. As a
result, any suitable metal and any suitable oxidizing agent may be
used in the invention. The metal may also be comprised of more than
one element, with the result that the deposition product may be
comprised of at least one oxidized species of more than one metal
element.

Preferably the metal is comprised of silver and the deposition product
is comprised of at least one oxidized species comprising silver. The
metal may, however, be further comprised of other metal elements such
as gold, copper, tin or zinc so that the deposition product is
comprised of at least one oxidized species comprising silver and one
or more other metals.

Where the metal is comprised of silver, the resulting deposition
product may exhibit significant antimicrobial properties. Without
intending to be limited by theory, it is believed that these
antimicrobial properties are due to the presence in the deposition
product of one or more oxidized silver species. The presence of other
metals in the deposition product may enhance these antimicrobial
properties or may provide other complementary properties to the
deposition product.

More particularly, it is believed that silver containing deposition
products produced using the methods of the invention may be comprised
of silver ions having valent states higher than one, such as for
example Ag (II) and Ag (III) valent states. It is also believed that
silver containing deposition products produced using the methods of
the invention may be comprised of silver ions having more than one
valent state so that the oxidized silver species may be comprised of a
multivalent substance. Finally, it is believed that silver containing
deposition products produced using the methods of the invention may be
comprised of a silver containing substance or a plurality of silver
containing substances which react over time to form other silver
containing substances which may exhibit differing antimicrobial
properties. It is believed that if this is the case, the deposition
products produced by the invention may be useful for providing a
varied antimicrobial response and for overcoming bacterial resistance.

In particular, in certain aspects, the methods of the invention may be
used to produce a deposition product which comprises a substance
having the general formula Ag.sub.7O.sub.8X, where X is an anion. The
deposition product may be further comprised of Ag.sub.2SO.sub.4. The
deposition product may also be comprised of other oxidized silver
compounds such as one or more silver oxides selected from the group of
silver oxides consisting of monovalent silver oxide, bivalent silver
oxide, trivalent silver oxide and mixtures thereof.

The anion X may be comprised of a single anion or may be comprised of
a plurality of different anions. The anion may therefore be comprised
of any anion or combination of ions. The anion may, for example, be
selected from the group of anions consisting of HCO.sub.3.sup.-,
CO.sub.3.sup.2-, NO.sub.3.sup.-, ClO.sub.4.sup.-, SO.sub.4.sup.2-,
F.sup.-, and mixtures thereof. The source of the anion may be a metal
compound which provides the ions of the metal. For example, where the
deposition solution is comprised of a silver salt such as silver
nitrate, the anion may be comprised of the nitrate ion (NO.sub.
3.sup.-). An alternative or secondary source of the anion X may
optionally be provided in order to provide sufficient quantities of
the anion for production of the deposition product. Where an
alternative or secondary source of the anion X is provided, the source
of anions may be comprised of any source, including but not limited to
any organic or inorganic acid.

Where the metal is comprised of silver, the composite materials
produced by the methods may therefore be useful as medical devices or
as components of medical devices due to their specific antimicrobial
properties. These composite materials may also provide other
advantages. As one example, the ability to provide a very thin coating
or layer of the deposition product on the substrate makes it possible
to minimize the amount of silver which must be used in the composite
material in order to provide a desired antimicrobial response. As a
second example, the ability to provide a very thin coating or layer of
the deposition product on the substrate minimizes the extent to which
the deposition product will interfere with the properties and
functions of the substrate, particularly if the deposition product is
deposited on the substrate so that it does not completely cover the
surface of the substrate. This second example may be particularly
significant where the substrate is comprised of an adhesive material
such as a skin adhesive layer.

In a first aspect, the invention is a method for producing a composite
material comprising a substrate and a deposition product, wherein the
deposition product is comprised of at least one oxidized species of a
metal, the method comprising the following steps: (a) first contacting
the substrate with a first basic environment comprising ions of the
metal in order to expose the substrate to the ions of the metal; and
(b) second contacting the substrate with a second basic environment in
order to produce the composite material.

The first basic environment may be comprised of any environment in
which metal ions are present under alkali conditions. The metal may be
comprised of any metal or combinations of metals but preferably the
metal is comprised of silver.

Preferably the first basic environment is comprised of a first basic
solution comprising an amount of a silver diamino complex. More
preferably, the first basic solution results from a mixture of a
silver compound and ammonium hydroxide in an aqueous medium.
Preferably the silver compound is selected from the group of silver
compounds consisting of silver salts, silver oxides and mixtures
thereof. More preferably the silver compound is comprised of silver
nitrate.

The first basic solution may have any alkaline pH. Preferably the
first basic solution has a pH in the range from about 8 to about 14.
Within these parameters, the amount of ammonium hydroxide in the first
basic solution is preferably selected such that a concentration of
ammonium hydroxide in the first basic solution is between about 25
percent and about 35 percent by volume of the first basic solution.
Preferably the amount of silver compound in the first basic solution
is selected such that a concentration of the silver compound in the
first basic solution is between about 1 gram per liter and about 20
grams per liter.

The second basic environment may be comprised of any environment
having alkali conditions. Preferably the second basic environment is a
strongly alkaline environment having a pH at least about 12.
Preferably the second basic environment is comprised of a second basic
solution containing an amount of a strong alkali compound. The strong
alkali compound may be comprised of any compound which can provide the
strong alkaline environment. For example, the strong alkali compound
may be comprised of one or more Group I elements, including lithium,
sodium, potassium, rubidium, cesium and francium. Preferably the
strong alkali compound is selected from the group of compounds
consisting of sodium hydroxide and potassium hydroxide and mixtures
thereof and more preferably the strong alkali compound is comprised of
sodium hydroxide. Preferably the amount of hydroxide compound in the
second basic solution is selected such that a concentration of the
hydroxide compound in the second basic solution is between about 15
grams per liter and about 35 grams per liter.

The first contacting step may be performed for any length of time
which is sufficient to expose the substrate to the ions of the metal.
Preferably the substrate is substantially completely exposed to the
ions of the metal. Preferably the first contacting step is performed
for between about 1 minute and about 10 minutes.

The second contacting step may be performed for any length of time
which is sufficient to cause the production of the deposition product.
Preferably the second contacting step is performed for a sufficient
time in order to maximize the yield of the deposition product.
Preferably the second contacting step is performed for between about 1
minute and about 60 minutes.

The first contacting step may be performed at any temperature. The
second contacting step may be performed at any temperature.
Preferably, however, the second contacting step is performed at a
temperature of between about 2 degrees Celsius and about 60 degrees
Celsius.

The method according to the first aspect may be further comprised of
the step of washing the composite material following the second
contacting step.

The method according to the first aspect may be further comprised of
the step of adding an amount of an oxidizing agent to the second basic
environment during the second contacting step. The oxidizing agent may
be comprised of any oxidizing agent which is compatible with the
metal, but the oxidizing agent is preferably selected from the group
of oxidizing agents consisting of persulfates, permanganates,
peroxides and mixtures thereof. More preferably the oxidizing agent is
comprised of a persulfate. The persulfate may be comprised of any
persulfate but preferably the persulfate is selected from the group of
persulfates consisting of potassium persulfate, sodium persulfate,
ammonium persulfate and mixtures thereof. More preferably the
persulfate is comprised of ammonium persulfate, potassium persulfate
or mixtures thereof, and most preferably the persulfate is comprised
of potassium persulfate.

The amount of the oxidizing agent is preferably selected to be
compatible with the amount of the ions of the metal so that the
deposition product can be produced as efficiently as possible. In
other words, the amount of the oxidizing agent is preferably selected
to be a stoichiometrically appropriate amount relative to the amount
of the ions of the metal. Preferably the amount of persulfate
oxidizing agent is selected such that a concentration of the
persulfate in the second basic solution is between about 1 gram per
liter and about 25 grams per liter.

The method according to the first aspect may be further comprised of
the step, prior to the first contacting step, of etching the substrate
by immersing the substrate in an etching solution in order to prepare
the substrate for the deposition product. The etching step may involve
either or both of a physical process or a chemical process. The
etching step preferably prepares the substrate for the deposition
product by increasing the roughness of the substrate surface and/or
creating attraction sites for adsorption and/or deposition of the
deposition product.

Any etching solution may be utilized which is suitable for a
particular substrate. For example, where the substrate is comprised of
an organic material or polymer such as polyethylene, the etching
solution is preferably comprised of a mixture of an alcohol and an
aqueous solution of a hydroxide compound. The hydroxide compound may
be comprised of any hydroxide compound but is preferably selected from
the group of hydroxide compounds consisting of sodium hydroxide,
potassium hydroxide and mixtures thereof. More preferably the
hydroxide compound is comprised of sodium hydroxide. The etching step
may be performed for any length of time sufficient to prepare the
substrate, but preferably the etching step is performed for less than
about 20 minutes and preferably is performed for at least 5 minutes.

The method according to the first aspect may be further comprised of
the step of adding a residual silver compound to the second basic
environment during the second contacting step. The residual silver
compound may be comprised of any suitable source of silver ions, but
preferably the residual silver compound is comprised of silver
nitrate. Preferably the amount of residual silver compound is selected
such that a concentration of the residual silver compound in the
second basic solution is between about 1 gram per liter and about 5
grams per liter.

The method according to the first aspect may be further comprised of
the step of agitating the second basic environment during at least a
portion of the second contacting step in order to enhance the
production of the deposition product and the composite material.

In a second aspect, the invention is a method for producing a
deposition product, wherein the deposition product is comprised of at
least one oxidized species of a metal, the method comprising the
following steps: (a) providing a deposition solution comprising an
amount of ions of the metal and an amount of an oxidizing agent; and
(b) producing the deposition product by facilitating a chemical
reaction in the deposition solution between the ions of the metal and
the oxidizing agent.

The metal may be comprised of any metal or combinations of metals but
preferably the metal is comprised of silver so that the ions of the
metal are comprised of silver ions. The deposition solution may be
comprised of silver ions from any source or in any form but preferably
the deposition solution is comprised of an aqueous solution of a
silver salt. More preferably the silver salt is comprised of silver
nitrate.

The ions of the metal may be present in any concentration. Preferably,
where the ions of the metal are comprised of silver ions, the amount
of the silver ions is selected so that a concentration of the silver
salt in the deposition solution is between about 1 gram per liter and
about 20 grams per liter.

The oxidizing agent may be comprised of any oxidizing agent which is
compatible with the metal, but the oxidizing agent is preferably
selected from the group of oxidizing agents consisting of persulfates,
permanganates, peroxides and mixtures thereof. More preferably the
oxidizing agent is comprised of a persulfate. The persulfate may be
comprised of any persulfate but preferably the persulfate is selected
from the group of persulfates consisting of potassium persulfate,
sodium persulfate, ammonium persulfate and mixtures thereof. More
preferably the persulfate is comprised of ammonium persulfate,
potassium persulfate or mixtures thereof, and most preferably the
persulfate is comprised of potassium persulfate.

The amount of the oxidizing agent is preferably selected to be
compatible with the amount of the ions of the metal so that the
deposition product can be produced as efficiently as possible. In
other words, the amount of the oxidizing agent is preferably selected
to be a stoichiometrically appropriate amount relative to the amount
of the ions of the metal. For example, where the metal is comprised of
silver nitrate the amount of silver nitrate is preferably selected
such that a concentration of the silver nitrate in the deposition
solution is between about 1 gram per liter and about 20 grams per
liter, in which case the amount of the oxidizing agent is preferably
selected so that a concentration of the oxidizing agent in the
deposition solution is between about 1 gram per liter and about 50
grams per liter.

The method according to the second aspect may be used to produce a
deposition product which comprises a substance having the general
formula Ag.sub.7O.sub.8X, where X is an anion. The deposition product
may be further comprised of Ag.sub.2SO.sub.4. The deposition product
may also be comprised of other oxidized silver compounds such as one
or more silver oxides selected from the group of silver oxides
consisting of monovalent silver oxide, bivalent silver oxide,
trivalent silver oxide and mixtures thereof.

The anion X may be comprised of a single anion or may be comprised of
a plurality of different anions. The anion may therefore be comprised
of any anion or combination of ions. The anion may, for example, be
selected from the group of anions consisting of HCO.sub.3.sup.-,
CO.sub.3.sup.2-, NO.sub.3.sup.-, ClO.sub.4.sup.-, SO.sub.4.sup.2-,
F.sup.-, and mixtures thereof. The source of the anion may be a metal
compound which provides the ions of the metal. For example, where the
deposition solution is comprised of a silver salt such as silver
nitrate, the anion may be comprised of the nitrate ion (NO.sub.
3.sup.-). An alternative or secondary source of the anion X may
optionally be provided in order to provide sufficient quantities of
the anion for production of the deposition product.

As a result, in the method according to the second aspect, the method
may be further comprised of the step of adding a source of anions to
the deposition solution. The source of anions may be comprised of one
or more acids. The acid may be comprised of any organic or inorganic
acid. For example, the acid may be selected from the group of acids
consisting of carbonic acid, nitric acid, perchloric acid, sulfuric
acid, acetic acid, fluoroboric acid, phosphoric acid, phosphorous
acid, citric acid, acetylsalicylic acid and mixtures thereof. The
amount of the source of anions which is added to the deposition
solution preferably is an amount which is selected to be compatible
with the amount of the ions of the metal. In other words, the amount
of the source of anions is preferably selected to be a
stoichiometrically appropriate amount relative to the amount of the
ions of the metal.

The deposition product producing step is preferably performed at a
relatively low temperature, since the deposition product may
experience increasing solubility with increasing temperature. The
deposition product producing step is preferably performed at a
temperature of between about 2 degrees Celsius and about 60 degrees
Celsius, more preferably at a temperature of between about 2 degrees
Celsius and about 40 degrees Celsius, and even more preferably at a
temperature of between about 10 degrees Celsius and about 25 degrees
Celsius.

Preferably the deposition solution is agitated during at least a
portion of the deposition product producing step in order to enhance
the production of the deposition product.

The method according to the second aspect may be used to produce the
deposition product as a product, or may be used to produce a composite
material comprising a substrate and the deposition product. Where the
method is used to produce a composite material, the method may be
further comprised of the following steps: (a) providing a substrate;
and (b) contacting the substrate with the deposition solution during
the deposition product producing step, thereby producing a composite
material comprising the substrate and the deposition product.

The substrate contacting step may be performed for any length of time
which is sufficient to produce the composite material having a desired
composition. The substrate contacting step is preferably performed for
at least about 1 minute, more preferably for between about 1 minute
and about 60 minutes, even more preferably for between about 1 minute
and about 20 minutes, and even more preferably for between about 2
minutes and about 10 minutes.

The method in the second aspect may be further comprised of the step,
following the substrate contacting step, of washing the composite
material.

The method according to the second aspect may be further comprised of
the step, prior to the substrate contacting step, of etching the
substrate by immersing the substrate in an etching solution in order
to prepare the substrate for the deposition product. The etching step
may involve either or both of a physical process or a chemical
process. The etching step preferably prepares the substrate for the
deposition product by increasing the roughness of the substrate
surface and/or creating attraction sites for adsorption and/or
deposition of the deposition product.

Any etching solution may be utilized which is suitable for a
particular substrate. For example, where the substrate is comprised of
an organic material or polymer such as polyethylene, the etching
solution is preferably comprised of a mixture of an alcohol and an
aqueous solution of a hydroxide compound. The hydroxide compound may
be comprised of any hydroxide compound but is preferably selected from
the group of hydroxide compounds consisting of sodium hydroxide,
potassium hydroxide and mixtures thereof. More preferably the
hydroxide compound is comprised of sodium hydroxide. The etching step
may be performed for any length of time sufficient to prepare the
substrate, but preferably the etching step is performed for less than
about 20 minutes and preferably is performed for at least 5 minutes.
Where the etching step is performed, the method according to the
second aspect preferably further comprises the step, following the
etching step, of washing the substrate to remove residual alkali from
the substrate.

The method according to the second aspect may be further comprised of
the step, following the substrate contacting step, of immersing the
composite material in boiling water. The immersing step may be useful
for converting the deposition product into other oxidized silver
species (such as silver oxides), thus potentially providing an
opportunity further to "engineer" the composite material to provide
desired properties of the deposition product. The immersing step may
be performed for any length of time, but preferably the immersing step
is performed for at least about 1 minute.

The composite material may be produced for many different applications
including for electronics, materials engineering and medical purposes.
The method according to the second aspect is particularly suited for
the production of medical devices in circumstances where the metal is
silver and the deposition product is comprised of an oxidized silver
species having the general formula Ag.sub.7O.sub.8X and optionally
Ag.sub.2SO.sub.4 and/or optionally one or more silver oxide compounds,
due to the antimicrobial properties exhibited by the deposition
product and to the capability to control the extent of the deposition
of the deposition product on the substrate.

The term "medical device" as used herein means any article which has a
medical application where antimicrobial properties may be desirable,
and includes all natural and synthetic materials and both fibrous and
non-fibrous materials. For example, the materials may be comprised of
a metal, plastic, paper, glass, ceramic, textile, rubber, polymer,
composite material or any other material or combination of materials.
Non-limiting examples of medical devices which are encompassed by the
invention include wound dressings, splints, sutures, catheters,
implants, tracheal tubes, orthopedic devices, drains, shunts,
connectors, prosthetic devices, needles, medical instruments,
laboratory, clinic and hospital equipment, furniture and furnishings,
dental devices, as well as health care products such as personal
hygiene products, sterile packaging, clothing, footwear etc.

Accordingly, the composite material may comprise a medical device or a
component of a medical device and the term "medical device" as used
herein extends to both medical devices and components of medical
devices.

In a preferred embodiment, the substrate is comprised of a wound
dressing. The wound dressing may be comprised of any material or
combination of materials, including but not limited to metals,
ceramics, glass, polymers, plastics, composite materials, natural
materials, synthetic materials, synthetic textiles such as HDPE,
rayon, nylon, polyacetates, polyacrylics and glass and natural
textiles such as cellulose, wool, jute and cotton, whether in fibrous
or non-fibrous form.

In a preferred embodiment of wound dressing, the wound dressing may be
comprised of a polymer material such as high density polyethylene and
may be further comprised of an adhesive material comprising a skin
adhesive layer. The skin adhesive layer may be comprised of a cross-
linked silicon gel material. The wound dressing and/or the cross-
linked silicon gel material may for example be comprised of a product
sold under the Mepitel.TM. trade-mark or the Safetac.TM. trade-mark,
both of which trade-marks are owned by Molnlycke Health Care AB of
Sweden.

In one application, the deposition product may be selectively
deposited on the skin adhesive layer and the production of the
deposition product is preferably controlled so that the deposition
product does not materially interfere with the adhesive properties of
the skin adhesive layer, yet still provides an acceptable
antimicrobial effect without significant undesirable toxic effects.
This result may be achieved by depositing the deposition product on
the skin adhesive layer such that the deposition product provides a
desired antimicrobial effect but does not completely cover the surface
of the skin adhesive layer. In this application, preferably the amount
of the deposition product which is deposited on the substrate is such
that the amount of total silver on the substrate is selected to be
between about 0.1 mg/cm.sup.2 and about 1.0 mg/cm.sup.2, or more
preferably between about 0.2 mg/cm.sup.2 and about 0.6 mg/cm.sup.2, in
order to achieve the desired result.

In other applications in which the deposition product is not deposited
on an adhesive such as the skin adhesive layer, the amount of the
deposition product is preferably controlled to balance the desired
antimicrobial effect, undesirable toxic effects, and economic
considerations.

In a third aspect, the invention is a medical device comprising a
composite material, wherein the composite material is comprised of a
substrate and a deposition product and wherein the deposition product
is comprised of an antimicrobially active oxidized silver species
comprising a silver salt and a silver oxide.

The medical device according to the third aspect may be produced using
any of the methods of the invention. Preferably the medical device is
produced using a method according to the second aspect of the
invention.

In certain preferred embodiments the invention provides methods for
depositing a deposition product comprising at least one oxidized
silver species onto a substrate, thus producing a composite material.
Since the oxidized silver species of the invention exhibit an
antimicrobial activity, composite materials comprising the oxidized
silver species can be used in various medical devices for prevention
or inhibition of infections. These medical devices may include but are
not limited to wound dressings, adhesives, sutures, catheters and
other articles where antimicrobial properties are desirable.

The preferred embodiments of the invention may be used to produce
deposition products and composite materials from aqueous solutions
under a wide range of pH conditions, involving reactions in either
acidic or alkaline solutions. The methods can be performed at, but are
not limited to, temperatures between about 2 degrees Celsius and about
60 degrees Celsius with about 10 degrees Celsius to about 40 degrees
Celsius being the most preferable.

The method steps for certain preferred embodiments of the invention
are as follows:

I. Under Acidic Conditions:

(a) immersing an article to be used as a medical device in an aqueous/
alcohol solution of NaOH for a sufficient time to provide a reasonable
etching and cleaning of the surface, followed by washing of the
article with distilled water until a pH of 7 is attained, in order to
remove residual alkali; (b) immersing the article in an aqueous silver
salt solution. The aqueous silver salt solution may be prepared from
any silver salt which is soluble in water with the most preferred
silver salt being silver nitrate; (c) adding a stoichiometrically
suitable quantity of an oxidizing agent to the mixed silver salt
solution containing the article. The oxidizing agent can be any
oxidizing substance such as persulfates, permanganates, hydrogen
peroxide and the like, with potassium persulfate (K.sub.2S.sub.2O.sub.
8) being the most preferred oxidizing agent; (d) adding a
stoichiometrically suitable quantity of an acid to the mixed silver
salt solution containing the immersed article in order to provide a
source of anions. The acids that can be used include any inorganic or
organic acids including, but not limited to carbonic acid, nitric
acid, perchloric acid, sulfuric acid, acetic acid, fluoroboric acid,
phosphoric acid, phosphorous acid, citric acid, acetylsalicylic acid
and mixtures thereof, but most preferably nitric acid, perchloric
acid, phosphoric acid, acetic acid or sulfuric acid; (e) agitating the
article in the mixed silver salt solution comprising the soluble
silver salt (preferably AgNO.sub.3), the acid (preferably nitric acid,
perchloric acid, phosphoric acid, acetic acid or sulfuric acid), and
the oxidizing agent (preferably potassium persulfate) at temperatures
between 2 degrees Celsius and 30 degrees Celsius with temperatures
between 10 degrees Celsius and 25 degrees Celsius being the most
preferred for between about 2 and 40 minutes until the article is
coated with a grayish, gray or black color; (f) removing the article
from the slurry and washing the article with distilled water until a
pH of 7 is achieved; and (g) drying the article at room temperature.

Alternatively after step (e) the article may be immersed in boiling
water (about 90 degrees Celsius to about 100 degrees Celsius) for at
least 1 minute.

II. Under Alkaline Conditions:

(a) immersing an article to be used as a medical device in an aqueous/
alcohol solution of NaOH for a sufficient time to provide a reasonable
etching and cleaning of the surface; (b) removing the article into a
solution containing a silver diamino complex in a concentration
sufficient to adsorb the silver ions at the surface of the article and
for a duration of about 2 minutes to about 5 minutes. The silver
diamino complex may be prepared by dissolving any silver salt or
silver oxide in ammonium hydroxide, and may be achieved by adding a
stoichiometrically suitable quantity of ammonium hydroxide to an
aqueous solution or suspension of the silver salt or silver oxide
until a clear colorless solution containing [Ag(NH.sub.3).sub.2].sup.+
is obtained. The pH of this solution is usually in the range from
about 8 to about 12; (c) removing the article without washing or
rinsing into another solution containing a strong alkali, most
preferably NaOH or KOH, and agitating the article in this solution
until a clear colorless solution is obtained and the article is
clearly dyed with a tan, gray, brown or black color, depending on the
desired amount of oxidized silver species. The time of contact of the
article with the alkaline solution may vary, depending on temperature
and silver ion concentration, but the most preferable duration is
about 1 minute to about 15 minutes at room temperature or about 1
minute to about 10 minutes at a temperature of between about 40
degrees Celsius and about 60 degrees Celsius; (d) removing the dyed
article from the solution and washing with distilled water until a pH
of 7 is achieved; and (e) drying the article at room temperature.

Alternatively, in step (c), the method may involve, depending on the
amount of silver required at the surface of the article, further
additions to the strong alkali solution of the silver diamino complex
solution and/or additions to the strong alkali solution of an
oxidizing agent such as a persulfate, permanganate, peroxide or a
mixture thereof, with potassium persulfate being the most preferred
oxidizing agent.

BRIEF DESCRIPTION OF DRAWINGS

Embodiments of the invention will now be described with reference to
the accompanying drawings, in which:

FIG. 1 is an XRD pattern generated from a deposition product obtained
from the reaction of AgNO.sub.3 and (NH.sub.4).sub.2S.sub.2O.sub.8
according to Examples 15-16.

FIG. 2 is an SEM micrograph (magnification=2000.times.) generated from
a deposition product obtained from the reaction of AgNO.sub.3 and
(NH.sub.4).sub.2S.sub.2O.sub.8 according to Examples 15-16.

FIG. 3 is an XRD pattern generated from a deposition product obtained
from the reaction of AgNO.sub.3 and K.sub.2S.sub.2O.sub.8 according to
Examples 15-16.

FIG. 4 is an SEM micrograph (magnification=2000.times.) generated from
a deposition product obtained from the reaction of AgNO.sub.3 and
K.sub.2S.sub.2O.sub.8 according to Examples 15-16.

FIG. 5(a) is an SEM micrograph (magnification=150.times.) generated
from a sample of uncoated HDPE mesh.

FIG. 5(b) is an SEM micrograph (magnification=1000.times.) generated
from a sample of HDPE mesh upon which a deposition product has been
deposited according to Examples 15-16.

FIG. 6(a) is a photograph depicting a controlled zone of inhibition
(CZOI) against Staphylococcus aureus for a sample of HDPE mesh coated
with a deposition product according to Examples 15-16.

FIG. 6(b) is a photograph depicting a controlled zone of inhibition
(CZOI) against Pseudomonas aeruginosa for a sample of HDPE mesh coated
with a deposition product according to Examples 15-16.

FIG. 6(c) is a photograph depicting a controlled zone of inhibition
(CZOI) against Candida albicans for a sample of HDPE mesh coated with
a deposition product according to Examples 15-16.

FIG. 7 is an SEM micrograph (magnification=30.times.) generated from a
substrate consisting of an uncoated sample of a perforated plastic
carrier material with a skin adhesive layer comprised of a hydrophobic
cross-linked silicon gel (trade-mark Mepitel.TM.).

FIG. 8 is an SEM micrograph (magnification=40.times.) generated from a
composite material consisting of a coated sample of a perforated
plastic carrier material with a skin adhesive layer comprised of a
hydrophobic cross-linked silicon gel (trade-mark Mepitel.TM.), in
which a relatively small amount of deposition product has been
deposited on the substrate in accordance with the second and third
aspects of the invention.

FIG. 9 is an SEM micrograph (magnification=2000.times.) generated from
the composite material of FIG. 8, depicting the density and coverage
of the deposition product on the substrate.

FIG. 10 is an SEM micrograph (magnification=40.times.) generated from
a composite material consisting of a coated sample of a perforated
plastic carrier material with a skin adhesive layer comprised of a
hydrophobic cross-linked silicon gel (trade-mark Mepitel.TM.), in
which a relatively larger amount of deposition product (relative to
FIG. 8 and FIG. 9) has been deposited on the substrate in accordance
with the second and third aspects of the invention.

FIG. 11 is an SEM micrograph (magnification=2000.times.) generated
from the composite material of FIG. 10, depicting the density and
coverage of the deposition product on the substrate.

FIG. 12 is an XRD pattern generated from a substrate consisting of an
uncoated sample of a perforated plastic carrier material with a skin
adhesive layer comprised of a hydrophobic cross-linked silicon gel
(trade-mark Mepitel.TM.).

FIG. 13 is an XRD pattern generated from a composite material
consisting of a coated sample of a perforated plastic carrier material
with a skin adhesive layer comprised of a hydrophobic cross-linked
silicon gel (trade-mark Mepitel.TM.), in which a deposition product
has been deposited on the substrate in accordance with the second and
third aspects of the invention.

FIG. 14 is a superimposition of the XRD patterns depicted in FIG. 12
and FIG. 13.

DETAILED DESCRIPTION

In preferred embodiments of the invention, antimicrobial properties of
medical devices are achieved by the adsorption and deposition of a
deposition product comprising an antimicrobially active silver species
within or at the surface of the medical device. These active silver
species may include but are not limited at all oxidized silver species
such as silver salts, silver oxide (Ag.sub.2O), higher silver oxides
i.e. Ag(II) and Ag(III) (AgO, Ag.sub.2O.sub.3, Ag.sub.3O.sub.4 or
like), silver oxy-salts with a general formula Ag.sub.7O.sub.8X where
X can include one of acid anions such as sulfates, chlorides,
phosphates, carbonates, citrates, tartrates, oxalates and like. The
deposition product may also contain some elemental silver deposited
during the process.

The term "total silver" as used in herein is the total amount of
silver as determined by a chemical analysis, which may include
elemental (metallic) silver as well as silver originating from
oxidized silver species.

The term "oxidized silver species" as used herein may involve but is
not limited at all compounds of silver where said silver is in +I, +II
or +III valent states or any combinations thereof. These oxidized
silver species include, for example silver (I) oxide, silver (II)
oxide, silver (III) oxide or mixtures thereof, all silver salts having
a solubility product higher than 10.sup.-20 (such as for example
Ag.sub.2SO.sub.4, AgCl, Ag.sub.2S.sub.2O.sub.8, Ag.sub.2SO.sub.3,
Ag.sub.2S.sub.2O.sub.3, Ag.sub.3PO.sub.4, and the like), and silver
oxy-salts such as Ag.sub.7O.sub.8X were X can include but is not
limited at NO.sub.3.sup.-, ClO.sub.4.sup.-, SO.sub.4.sup.2-, F.sup.-
etc.

The term "medical device materials" as used herein may include
materials such as metals, ceramics, glass, polymers, plastics,
composite materials, a variety of natural materials, fabrics, textile
made of either synthetic (HDPE, rayon, nylon, polyacetates,
polyacrylics, glass etc.) or natural (cellulose, wool, jute, cotton,
etc.) fibers.

The term "bacteriostatic activity", as used herein relates to the
inhibition of bacterial growth, but not to actually killing the
bacteria. Successful treatment therefore requires the host's immune
system to clear the pathogen. Treatment is compromised when the
antimicrobial materials are stopped before the pathogen has been
completely cleared.

The term "bactericidal activity" as used herein relates to killing
bacteria with or without lysis of the target cell. These types of
antimicrobial materials are particularly advantageous in
immunosuppressed individuals. A disadvantage to bactericidal activity
is cell lysis, which can release lipolysaccharides which are toxic to
the host. However, if the concentration of the said antimicrobial
material is relatively low so that toxic effects cannot occur, a
combination of both bacteriostatic and bactericidal activities may be
ideal for antimicrobial materials.

In the preferred embodiments, the deposition of the deposition product
comprising the oxidized silver species is accomplished by first
providing an aqueous solution of monovalent silver salt or a silver
complex such as silver nitrate, perchlorate or silver diamino complex,
with silver nitrate being the most preferable if the reaction is
carried out under acidic conditions or at close to neutral conditions
(i.e. at pH below 7), and with silver diamino complex, (i.e.,
[Ag(NH.sub.3).sub.2].sup.+) being the most preferable if the reaction
is carried out under alkaline conditions (i.e. at pH above 7).

Prior to the production of the composite material comprising the
article as a substrate and the deposition product, the article is
preferably immersed in an alkaline solution containing 50 vol. %
ethanol and 50 vol. % of an aqueous solution containing 30 g/L NaOH.
Other cleaning and etching solutions can be used depending upon the
material from which the medical device is made, upon the toxicity of
the said cleaning or etching solutions, and upon the possibility that
some toxic substances may adsorb at the surface of the article. Of
course any use of toxic or carcinogenic substances during the etching
step should be avoided. If production of the deposition product is
carried out under acidic conditions, the article is preferably washed
with distilled water after the etching step until a pH of 7 is
achieved in order to remove residual alkali remaining after the
etching step.

When the reaction is carried out in the pH range below 7 (i.e., under
acidic conditions), the clean pretreated article to be used as a
medical device containing oxidized silver species at the surface of
the same is simply immersed into an agitated 1% AgNO.sub.3 aqueous
solution as a deposition solution. After exposure of the said article
to the deposition solution for a duration preferably of about 2 to
about 10 minutes, a solution of an oxidizing agent is added.
Alternatively, the oxidizing agent may be added to the deposition
solution before the article is immersed into the deposition solution,
but this may result in some production of the deposition product
before the article is present in the deposition solution.

Although a wide range of oxidizing agents such as permanganates,
persulfates, hydrogen peroxide, hypochlorites etc., may be used under
specific conditions and with the proper concentrations, the preferred
oxidizing agent is a persulfate, more preferably either ammonium
persulfate or potassium persulfate., and most preferably potassium
persulfate The persulfate facilitates the precipitation and deposition
of the deposition product on or within the article.

The concentration of persulfate in the deposition solution may be in a
range from about 1 gram per liter to about 250 gram per liter with the
concentration of about 50 gram per liter being the most preferable.
After agitation for about 2 minutes to about 5 minutes, the solution
of 1% AgNO.sub.3 and persulfate may be acidified with an organic or
inorganic acid such as HNO.sub.3, HClO.sub.4, H.sub.2SO.sub.4 or
CH.sub.3COOH such that the concentration of the free acid preferably
is about 9% HNO.sub.3, 9% HClO.sub.4 acid, 5% H.sub.2SO.sub.4, or 5%
CH.sub.3COOH. Although other acids may be used the most preferable
acids are H.sub.2SO.sub.4, HClO.sub.4 or HNO.sub.3.

The agitation of the deposition solution is not strictly required, but
in order to achieve a more uniform distribution of the deposition
product and an efficient reaction yield, the agitation of the solution
is recommended. Agitation can be realized by many different ways such
as for example mechanical stirring, magnetic stirring or ultrasonic
agitation.

Following addition of the persulfate (preferably potassium persulfate)
to the deposition solution of 1% AgNO.sub.3 within the time of about 1
minute to about 10 minutes, and depending on the concentration of the
persulfate as well as on the conditions of agitation, the formation
first of a yellow brown color of the solution and then a black grayish
precipitate will occur. This brown color of the solution is attributed
to the oxidation of Ag(I) to Ag(II).

The black grayish deposit at the article or in the bulk solution is a
consequence of the formation of silver oxy-salts such as Ag.sub.7O.sub.
8X, were X is an anion, depending on the acid used in the method e.g.
HNO.sub.3 (NO.sub.3-), H.sub.2SO.sub.4 (SO.sub.4.sup.2-), etc. The
decomposition of the silver oxy-salts may be presented as: Ag(Ag.sub.
3O.sub.4).sub.2X.dbd.AgX+AgO (1)

Persulfates are powerful oxidizing agents. They can therefore be
reduced in aqueous solutions according to the following reactions:
S.sub.2O.sub.8.sup.2-+2e.sup.-=2SO.sub.4.sup.2-,with E.sup.0=1.96 V
(2) S.sub.2O.sub.8.sup.2-+2H.sup.++2e.sup.-=2HSO.sub.4.sup.-, with
E.sup.0=1.96 V (3) and S.sub.2O.sub.8.sup.2-+2H.sub.2O=2H.sup.+
+2SO.sub.4.sup.2-+H.sub.2O.sub.2, with .DELTA.G.sup.0=-36 kJ/mol (4)

A consequence of the reduction of persulfate is the oxidation of Ag(I)
to Ag(II) and Ag(III), probably according to the following reactions:
Ag.sup.+=Ag.sup.2++e.sup.-, with E.sup.0=1.98 V (5) Ag.sup.++H.sub.
2O.dbd.AgO.sup.++2H.sup.++e.sup.-, with E.sup.0=1.998 V (6) Ag.sup.2+
+H.sub.2O.dbd.AgO.sup.++2H.sup.++e.sup.-, with E.sup.0=2.06 V (7)
Ag.sup.++H.sub.2O.dbd.AgO+2H.sup.++e.sup.-, with E.sup.0=1.772 V (8)

In this way the composite material comprising the article to be used
as a medical device and the deposition product may include a
combination of oxidized silver species i.e. Ag(I)- and Ag(II)-oxides
as well as silver salts such as nitrates, persulfates, sulfates,
phosphates, perchlorates and like, silver salts of a general formula
Ag.sub.7O.sub.8X and perhaps traces of pure elemental silver. After
production of the composite material, the article is removed from the
deposition solution and then preferably washed with distilled water
until a pH of 7 is achieved. When the washing is completed, the
medical device comprising the composite material may be dried at room
temperature and packaged.

When the reaction is carried out in the pH range above 7 (i.e., under
alkaline conditions) the article to be used as a medical device is
first immersed in an etching solution comprising an alkaline solution
containing alcohol. The most preferable solution according to this
invention is either NaOH or KOH with concentrations 15 to 40 g/L. The
alcohol used in this solution may be ethyl alcohol, methyl alcohol or
mixtures therein in a concentration above 50 vol. %. The immersion of
the article into the etching solution is carried out in order to etch
and clean the surface of the article to provide a reasonable adhesion
of the deposition product comprising an oxidized silver species which
is deposited on or within the article thereafter. The immersion time
of the article is preferably in the range of between about 5 minutes
and about 20 minutes, with about 10 minutes being the most preferable.

After the exposure to the alkali/alcohol solution for about 10
minutes, the article is then removed without washing or rinsing into a
first basic environment comprising a first basic solution containing
silver diamino complex i.e. [Ag(NH.sub.3).sub.2].sup.+ in a
concentration sufficient to adsorb silver ions at the surface of the
article and for a duration of about 2 minutes to about 5 minutes. The
silver diamino complex is preferably prepared from a silver salt or
silver oxide dissolved or suspended in water by a dissolution with
NH.sub.4OH (28 vol. %).

Consequently, the first basic solution is prepared in a way such that
a solution of any silver salt (such as for example AgNO.sub.3 or
AgClO.sub.4) or any silver oxide (such as Ag.sub.2O or Ag.sub.2O.sub.2
or AgO) or any silver salt suspended in water (such as AgCl, Ag.sub.
2CO.sub.3, Ag.sub.2SO.sub.4 or the like), the ammonium hydroxide is
added in a stoichiometrically suitable concentration so that a clear
colorless solution is obtained. The concentration of silver ion in
this silver diamino complex solution, as calculated for Ag.sup.+ ion
can vary from 1 to 20 g/L with about 10 g/L being the most preferable.
The pH of the first basic solution is usually between about 8 and
about 12 with the most preferred pH being in the range of between
about 10 and about 11.

After exposure of the article to the first basic solution for about 2
minutes to about 5 minutes, the article is removed without washing or
rinsing into a second basic environment comprising a second basic
solution containing a strong alkali, most preferably NaOH or KOH. The
article is kept in this solution under agitation until a clear
colorless solution is obtained and the article is dyed with a tan,
gray, brown or black color, depending on the desired amount of
oxidized species to be deposited at or within the surface of the
article. The time of contact of the article with the second basic
solution may vary depending on temperature and the silver ion
concentration, but most preferable time is about 1 minute to about 15
minutes at room temperature or about 1 minute to about 10 minutes at a
temperature of between about 40 degrees Celsius and about 60 degrees
Celsius.

Alternatively, the method may involve an addition of an oxidizing
agent to the second basic solution, preferably a persulfate, more
preferably either ammonium persulfate or potassium persulfate, and
most preferably potassium persulfate. The oxidizing agent may be added
directly to the second basic solution containing the article. In
addition, depending on the amount of silver desired to be deposited as
the deposition product, addition of a residual silver compound such as
the silver diamino complex [Ag(NH.sub.3).sub.2].sup.+ may also be
beneficial.

Upon immersion of the article, previously exposed to the first basic
solution, into the second basic solution, the following reaction at
the surface of the article may occur: 2Ag(NH.sub.3).sub.2NO.sub.
3+2NaOH.dbd.Ag.sub.2O+4NH.sub.3+H.sub.2O+2NaNO.- sub.3 (9)

In this way, at the surface of the article, Ag.sub.2O will deposit as
the result of the reaction (9). The addition of an oxidizing agent
such as ammonium persulfate (i.e., (NH.sub.4).sub.3S.sub.2O.sub.8) to
the second basic solution may result in the oxidation of silver ions
and the reduction of S.sub.2O.sub.8.sup.2- ions pursuant to the
following reactions: Ag.sup.+=Ag.sup.2++e.sup.-, with E.sup.0=1.96 V
(10) and S.sub.2O.sub.8.sup.2-+2e.sup.-=2SO.sub.4.sup.2-, with E.sup.
0=1.96 V (11)

The reactions of Ag(NH.sub.3).sub.2.sup.+ ion with ammonium persulfate
can be represented as follows: Ag(NH.sub.3).sub.2NO.sub.3+(NH.sub.
4).sub.2S.sub.2O.sub.8=Ag.sub.2S.sub.2- O.sub.8+2NH.sub.4NO.sub.
3+4NH.sub.3 (12) Ag.sub.2S.sub.2O.sub.8+H.sub.2O=2AgO+2H.sub.2SO.sub.4
(13) Ag(NH.sub.3).sub.2NO.sub.3+(NH.sub.4).sub.2S.sub.2O.sub.8+2H.sub.
2O=2NH.s- ub.4NO.sub.3+2AgO+2H.sub.2SO.sub.4+4NH.sub.3 (14) or
Ag(NH.sub.3).sub.2NO.sub.3+(NH.sub.4).sub.2S.sub.2O.sub.8+2H.sub.
2O=2NH.s- ub.4NO.sub.3+2AgO+2(NH.sub.4).sub.2SO.sub.4 (15)

In this way, the deposition product may contain Ag.sub.2O, AgO or
other higher oxides of silver Ag(II), Ag(III) and mixtures therein.
Also, if alcohol is present in the reacting solution, due to
transferring from the etching solution some elemental silver may occur
in the deposition product. This is because in the presence of
persulfates, alcohols can be oxidized to aldehydes according to the
reactions: CH.sub.3OH.dbd.H.sub.2CO+2H.sup.++2e.sup.- (16) C.sub.
2H.sub.5OH.dbd.CH.sub.3CHO+2H.sup.++2e.sup.- (17)

Under the alkaline conditions, the aldehydes can reduce the silver
ions to the elemental silver according to the reaction: 2Ag(NH.sub.
3).sub.2OH+HCHO=2Ag+4NH.sub.3+HCOOH+H.sub.2O (18)

After production of the composite material comprising the article and
the deposition product comprising the oxidized silver species is
completed, the article is removed, carefully washed with water until a
pH of 7 is achieved. The article may then be dried at room temperature
and packaged.

Following are examples which illustrate the present invention.

EXAMPLES

Example 1

Nine (9) pieces of high density polyethylene mesh (HDPE), with
dimensions 10 .times.8 cm each, were immersed in 100 ml of an etching
solution containing 50 mL alcohol (95% C.sub.2H.sub.5OH and 5% CH.sub.
3OH) and 50 mL of 28 g/L NaOH solution for 5 minutes. After 5 minutes
of etching the HDPE mesh was transferred without washing or rinsing
into 40 mL of an Ag.sup.+ solution, containing 15.3 g/L AgNO.sub.3 and
a stoichiometrically suitable volume of NH.sub.4OH (28 vol. %). The
HDPE mesh was kept in this solution for 2 minutes. After 2 minutes of
exposure to the ammoniacal Ag(NH.sub.3).sub.2.sup.+ solution, the HDPE
mesh was transferred without washing or rinsing into 150 mL of a 28 g/
L NaOH solution stirred with a magnetic stirrer. As soon as the HDPE
mesh was immersed into NaOH solution, the formation of a precipitate
yellowish-brown in color occurred. Under agitation a residual silver
compound (about 38 mL of the Ag(I) solution) was added and after that
5 mL of a 250 g/L (NH.sub.4).sub.2S.sub.2O.sub.8 solution was added.
Agitation was continued for 10 minutes. During this time the solution/
precipitate became black. The HDPE mesh was uniformly coated and was
black and shiny in appearance. The coated HDPE mesh was then removed
from the solution and carefully washed with distilled water until pH
7.00, and dried at room temperature. After drying, the mesh was a
black and shiny in appearance.

Chemical analysis determined that the HDPE mesh coated with oxidized
silver species contained about 0.08 mg total silver per cm.sup.2 of
mesh. The coated mesh was further analyzed by XRD analysis. As found
by the XRD analysis the mesh included Ag.sub.2O, Ag(II) oxides, Ag.sub.
7O.sub.8NO.sub.3 and some traces of the elemental silver. Both
bacteriostatic and bactericidal activities of silver coated HDPE
substrates were tested against Pseudomonas aeruginosa and
Staphylococcus aureus. One hour bactericidal activity tests of coated
HDPE mesh against both Pseudomonas aeruginosa and Staphylococcus
aureus were positive. The bacteriostatic activity was also tested. The
controlled zone of inhibition surrounding the test sample, where no
bacteria growth occurred, was estimated at about 9 mm to about 10 mm.

Example 2

Samples of HDPE mesh with dimensions 10.times.8 cm were immersed in
100 mL of an etching solution containing 50 mL of 28 g/L NaOH and 50
mL of denatured ethanol (95% C.sub.2H.sub.5OH and 5% CH.sub.3OH) for 5
minutes. After 5 minutes of etching the HDPE mesh was transferred
without washing or rinsing into 40 mL of an ammoniacal Ag(I) solution
containing 15.3 g/L AgNO.sub.3 and a stoichiometrically suitable
quantity of NH.sub.4OH (28 vol. %). The HDPE mesh was kept in this
solution for 2 minutes. The HDPE mesh was then transferred without
washing or rinsing into 150 mL of a solution containing 28 g/L NaOH.
The NaOH solution immediately became brown. Upon addition of a
residual silver compound (about 38 mL of the Ag(I) solution) the
solution turned to a dark brown color and with a continued agitation
for about 5 minutes the solution became black. When the agitation was
stopped, the black precipitate occurred in the bulk solution as a
result of its separation from the HDPE mesh material. After washing
and rinsing with distilled water the mesh appeared to be light tan or
at the most slightly gray as a consequence of the coating with silver
compounds.

The amount of total silver deposited on the HDPE mesh as determined by
chemical analysis was estimated at about 0.04 mg/cm.sup.2.
Antimicrobial activities (bactericidal and bacteriostatic) were tested
against Pseudomonas aeruginosa and Staphylococcus aureus. One hour
bactericidal activity of the coated HDPE mesh was positive. The
bacteriostatic activity, as estimated according to the controlled zone
of inhibition (CZOI) for the bacterial growth was also positive. The
CZOI was estimated at about 4 mm.

Example 3

Samples of HDPE mesh were immersed in an etching solution containing
100 mL of 28 g/L NaOH solution for 5 minutes. The mesh was then
transferred without washing or rinsing into 40 mL of an ammoniacal
Ag(I) solution containing 15.3 g/L AgNO.sub.3 and a stoichiometrically
suitable volume of NH.sub.4OH (28%). After 2 minutes of immersion, the
mesh was transferred without washing or rinsing into 150 mL of a 28 g/
L NaOH solution stirred magnetically. The solution became immediately
brown due to formation of a precipitate. Addition of a residual silver
compound (about 38 mL of the Ag(I) solution) resulted in the formation
of a dark brown precipitate. The color of the solution did not change
further even after 30 minutes of mixing at room temperature. The HDPE
mesh was then washed and rinsed very carefully with distilled water.
The color of the HDPE mesh did not change significantly, but some
change in color from white to a light tan appeared.

The amount of total silver deposited on the HDPE mesh was estimated at
about 0.02 mg/cm.sup.2. The antimicrobial activities (both
bacteriostatic and bactericidal) of these samples were tested against
Pseudomonas aeruginosa and Staphylococcus aureus. The results showed a
positive bactericidal activity and the CZOI was estimated at about 3
mm.

Example 4

Samples of HDPE mesh were immersed in 100 mL of a solution containing
1 g AgNO.sub.3 and 1 mL of 67% HNO.sub.3 as a source of anions. After
5 minutes of immersion, 5 g of (NH.sub.4).sub.2S.sub.2O.sub.8
dissolved in 20 mL of water was added. The sample was left for 30
minutes at room temperature, during which the solution was stirred
occasionally with a glass rod. During this time the solution changed
color from colorless to a dark brown and a formation of a light gray
precipitate in the bulk solution appeared. After 30 minutes, the HDPE
mesh was removed from the solution and carefully washed with distilled
water. The washed HDPE mesh had a gray color. The coating was
uniformly distributed at the surface of this material.

The amount of total silver deposited on the HDPE mesh was estimated at
0.09 mg/cm.sup.2. The bactericidal activity for these samples was
positive. The CZOI was estimated at about 8 mm.

Example 5

HDPE mesh was coated with silver oxidized compounds using a method
similar to that described in Example 4, with a few differences as
outlined in the description that follows.

Samples of HDPE mesh were immersed in 100 mL of a solution containing
10 g/L AgNO.sub.3 and 15 mL/L HNO.sub.3 (67%) as a source of anions.
To this solution 10 mL of 500 g/L (NH.sub.4).sub.2S.sub.2O.sub.8 was
added. The solution was magnetically stirred. After 7 minutes of
stirring the solution became yellow-brown and formation of a very
small amount of precipitate occurred. The stirring was continued for
the next 30 minutes. After 30 minutes, the HDPE mesh was removed from
the slurry and carefully washed with distilled water. The washed HDPE
mesh had a gray color. The coating was uniformly distributed at the
surface of the HDPE mesh.

The amount of total silver deposited on the HDPE mesh was estimated at
0.08 mg/cm.sup.2. The bactericidal activities against Pseudomonas
aeruginosa and Staphylococcus aureus were positive. The CZOI was
estimated at about 7 mm.

Example 6

HDPE mesh was coated with silver oxidized compounds using a method
similar to that described in Example 4 and Example 5, with a few
differences as outlined in the description that follows.

Samples of HDPE mesh were immersed in 100 mL of a solution containing
10 g/L AgNO.sub.3 and 15 mL/L HNO.sub.3 (67%) as a source of anions.
To this solution 10 mL of 500 g/L (NH.sub.4).sub.2S.sub.2O.sub.8 was
added. The solution was agitated ultrasonically. After 2 minutes of
stirring the solution became yellow-brown and formation of a very
small amount of precipitate occurred. The stirring was continued for
the next 30 minutes. After 30 minutes, the HDPE mesh was removed from
the solution and carefully washed with distilled water. The washed
HDPE mesh had a gray color. The coating was uniformly distributed at
the surface of the HDPE mesh.

The amount of total silver deposited on the HDPE mesh was estimated at
0.08 mg/cm.sup.2. The bactericidal activities against Pseudomonas
aeruginosa and Staphylococcus aureus were positive. The CZOI was
estimated at about 7 mm.

Examples 7-9

In these examples the effect of different acids (i.e., sources of
anions) is clearly shown for coating of HDPE mesh with oxidized silver
species under acidic conditions. In Example 4, HNO.sub.3 was used as a
source of anions to supplement the anions contained in the AgNO.sub.3,
while in Examples 7-9 perchloric acid (HClO.sub.4), sulfuric acid
(H.sub.2SO.sub.4) and acetic acid (CH.sub.3COOH) respectively were
used as a source of anions.

Samples of HDPE mesh were immersed in 100 mL of a solution containing
1 g AgNO.sub.3. To this solution 1 mL of HClO.sub.4 (70%) (Example 7),
0.5 mL of H.sub.2SO.sub.4 (98%) (Example 8) and 15 mL of CH.sub.3COOH
(5%) (Example 9) were added. After 2 minutes of the exposure of HDPE
mesh to these solutions, 20 mL of 250 g/L (NH.sub.4).sub.2S.sub.2O.sub.
8 was added. The mixing was continued for the next 30 minutes. In the
solutions containing HClO.sub.4 (Example 7) and H.sub.2SO.sub.4
(Example 8) formation of a black grayish precipitate occurred similar
to Example 4. When the precipitate settled the solutions were clear
and yellow-brown in color. The yellow-brown color suggests the
presence of Ag(II) complexes in the solution. The coated HDPE mesh was
then removed from the slurry and carefully washed and rinsed with
distilled water and thereafter dried at room temperature. After drying
the HDPE mesh coated in the presence of 1 mL of HClO.sub.4 (70%)
(Example 7), or in the presence of 0.5 mL of H.sub.2SO.sub.4 (98%)
(Example 8) appeared to be grayish in color. However, the HDPE mesh
coated in the presence of 15 mL of CH.sub.3COOH (5%) (Example 9) was
white and it did not change its color.

The coated HDPE mesh (Examples 7-9) were analyzed for the total silver
content, and the antimicrobial activity was also evaluated against
Pseudomonas aeruginosa and Staphylococcus aureus. The amount of total
silver deposited on the HDPE mesh was estimated at 0.08 mg/cm.sup.2
(for samples coated in the presence of HClO.sub.4), 0.07 mg/cm.sup.2
(for samples coated in the presence of H.sub.2SO.sub.4) and 0.01 mg/
cm.sup.2 (for the samples coated in the presence of CH.sub.3COOH). The
bactericidal activities against Pseudomonas aeruginosa and
Staphylococcus aureus were positive. The CZOI was estimated at about 6
mm (for samples coated in the presence of HClO.sub.4 or H.sub.2SO.sub.
4) and about 1 to 2 mm (for samples coated in the presence of
CH3COOH).

Example 10

Samples of HDPE mesh with dimensions 10.times.8 cm were immersed in
100 mL of an etching solution containing 50 mL of 28 g/L NaOH and 50
mL of denatured ethanol (95% C.sub.2H.sub.5OH and 5% CH.sub.3OH) for 5
minutes. After 5 minutes of etching the HDPE mesh was transferred
without washing or rinsing into 40 mL of an ammoniacal Ag(I) solution
containing 15.3 g/L AgNO.sub.3 and a stoichiometrically suitable
quantity of NH.sub.4OH (28 vol. %). The HDPE mesh was kept in this
solution for 2 minutes. The HDPE mesh was then transferred without
washing or rinsing into 150 mL of a solution containing 28 g/L NaOH.
The NaOH solution immediately became brown. After mixing for 2
minutes, the solution became clear and colorless and the mesh was tan
in color. When the agitation was stopped, the HDPE mesh was removed
from solution and washed with distilled water. After washing and
rinsing the mesh appeared to be tan in color as a consequence of the
coating with silver compounds.

The coated HDPE mesh was analyzed for silver content and for
antimicrobial activity against Pseudomonas aeruginosa and
Staphylococcus aureus. These samples contained between 0.04 and 0.08
mg/cm total silver. The bactericidal activities against Pseudomonas
aeruginosa and Staphylococcus aureus were positive. The CZOI was
estimated at about 10 mm.

Example 11

A patterned wound dressing made of a perforated plastic carrier
material with a skin adhesive layer comprised of a hydrophobic cross-
linked silicon gel (trade-mark Mepitel.TM., product of Molnlycke
Health Care AB, Sweden), dimensions 8.times.15 cm was exposed to a
solution containing 15 g/L NaOH at room temperature for 5 minutes.
Under conditions of agitation 40 mL of a solution containing 15.3 g/L
AgNO.sub.3 and a proper volume of NH.sub.4OH (28 vol. %) was added.
The wound dressing was kept in this solution and agitated for the next
5 minutes. The wound dressing was then removed from the solution and
carefully washed with distilled water. Drops of water were removed
with a soft paper and the wound dressing was dried at room
temperature.

The coated wound dressing was analyzed for antimicrobial activity
against Pseudomonas aeruginosa and Staphylococcus aureus. MH plates
and Tryptic Soy Broth were used for analysis. Pseudomonas aeruginosa
standard was set to 0.5 McFarland standard. One hour of bactericidal
activity of the coated wound dressing against the bacteria where TSB
broths were incubated for 24 hours was positive. The controlled zones
of inhibition (CZOI), for the bacterial growth (bacteriostatic
activity) were above 8 mm. The same samples of coated wound dressing
were tested for seven days for antimicrobial activity. The values of
CZOI after 2 days were 20.5 mm, after 3 days 19 mm, after 4 days 20.5
mm, after 5 days 19 mm and after 7 days 7 mm. These results show very
good resistance towards bacteria for a relatively long time (7 days).

Example 12

A patterned wound dressing made of a perforated plastic carrier
material with a skin adhesive layer comprised of a hydrophobic cross-
linked silicon gel (trade-mark Mepitel.TM., product of Molnlycke
Health Care AB, Sweden), dimensions 8.times.15 cm was exposed to 500
mL of a 1% AgNO.sub.3 solution. To this solution was added 200 mL of a
solution containing 20 g K.sub.2S.sub.2O.sub.8 and mixing was
continuous for the next 20 minutes. The wound dressing was then
removed from the solution and carefully washed with distilled water.
Drops of water were removed with soft paper and the wound dressing was
dried at room temperature.

The coated wound dressing contained 0.25-0.55 mg/cm.sup.2 of total
silver. The coated wound dressing was then analyzed for antimicrobial
activity in the same manner as described in Example 11. The results
showed excellent antimicrobial activity for 7 days.

Example 13

A patterned wound dressing made of a perforated plastic carrier
material with a skin adhesive layer comprised of a hydrophobic cross-
linked silicon gel (trade-mark Mepitel.TM., product of Molnlycke
Health Care AB, Sweden), dimensions 8.times.15 cm was coated in a way
as described in Example 12, except that (NH.sub.4).sub.2 S.sub.2O.sub.
8 was used as an oxidizing agent instead of K.sub.2S.sub.2O.sub.8, in
the same amount and in the same manner as described in Example 12.

The coated wound dressing produced as described in this example was
analyzed for the antimicrobial activity. The results showed excellent
antimicrobial activity.

Example 14

A slurry was prepared by mixing 500 mL of a 1% AgNO.sub.3 solution and
200 mL of an aqueous solution containing 20 g K.sub.2S.sub.2O.sub.8
for 10 minutes. To this slurry a patterned wound dressing made of a
perforated plastic carrier material with a skin adhesive layer
comprised of a hydrophobic cross-linked silicon gel (trade-mark
Mepitel.TM., product of Molnlycke Health Care AB, Sweden), dimensions
of 8.times.15 cm was added and mixing was continued for the next 20
minutes. The coated wound dressing was then removed from the slurry,
carefully washed with water then dried as described in the Example 12.
The coated wound dressing was black-greyish in appearance.

The antimicrobial activity of the coated wound dressing was tested in
a way described in Example 11. The results showed excellent
antimicrobial activity for seven days.

Examples 15-16

All method steps were performed at room temperature (22 degrees
Celsius.+-.2 degrees Celsius), unless otherwise specified.

Samples of HDPE mesh were coated with oxidized silver species as
follows. HDPE mesh with dimensions 10.times.10 cm were immersed into
100 mL of a 1% AgNO.sub.3 solution and thoroughly wetted. After the
exposure of the HDPE mesh to the solution for 10 minutes, 20 mL of a
solution containing either 250 g/L of (NH.sub.4).sub.2S.sub.2O.sub.8
or 250 g/L of K.sub.2S.sub.2O.sub.8 was added under magnetic stirring.
The mixing was continued for the next 15 minutes. The coated HDPE mesh
was then removed from the slurry and was observed to be grayish-black
in appearance. After coating, the HDPE mesh was washed with water and
then dried.

The bacteriostatic activity for the controlled zone of inhibition
(CZOI) of bacterial or fungal growth was tested against Pseudomonas
aeruginosa, Staphylococcus aureus or Candida albicans, using standard
procedures as described in the literature.

Discussion of Examples 15-16

(a) Deposition of Silver Deposition Products Using (NH.sub.4).sub.
2S.sub.2O.sub.8

Upon addition of ammonium persulfate to the AgNO.sub.3 solution, a
gradual color change from colorless through yellow, brown and finally
to a cloud solution containing grayish-black precipitate was observed.
Time for the appearance of the grayish-black precipitate at room
temperature was estimated at 5 to 10 minutes. It was noted that if the
reaction takes place at temperatures above 30 degrees Celsius, the
precipitation and color change do not occur.

Persulfates are powerful oxidizing agents. In aqueous solutions
persulfates can be reduced to sulfates (S. I. Zhdanov, Sulfur,
Selenium, Tellurium and Polonium, in Standard Potentials in Aqueous
Solutions, A. J. Bard, R. Parsons and J. Jordan Editors, Marcel Dekker
Inc., New York (1985)). A consequence of the reduction of persulfate
is the oxidation of Ag(I) to Ag(II) and Ag(II) to Ag(III). The grayish-
black precipitate deposited on the HDPE mesh was formed as a result of
the reduction of persulfate and a consequent oxidation of Ag(I) ions.

During precipitation of the deposition product, the pH of the solution
dropped from about 2 to below 1. The decrease in pH of the solution
was more significant when K.sub.2S.sub.2O.sub.8 is used as an
oxidizing agent instead of (NH.sub.4).sub.2S.sub.2O.sub.8, in that a
decrease in pH from about 7 to below 1 was observed.

(b) Properties of Deposition Products Produced Using (NH.sub.4).sub.
2S.sub.2O.sub.8

The grayish-black precipitate itself represents a mixture of silver
argentic nitrate Ag(Ag.sub.3O.sub.4).sub.2NO.sub.3Ag.sub.7NO.sub.11
and Ag.sub.2SO.sub.4. Indeed, as found by XRD analysis, the peaks in
the patterns showed a reasonable match for Ag.sub.2SO.sub.4 and Ag.sub.
7O.sub.8NO.sub.3 (FIG. 1). It is apparent that the oxidation of
AgNO.sub.3 with (NH.sub.4).sub.2S.sub.2O.sub.8 leads to the
precipitation of silver oxy-salt Ag.sub.7NO.sub.11 and also Ag.sub.
2SO.sub.4. The precipitation of Ag.sub.2SO.sub.4 is usually not
observed when K.sub.2S.sub.2O.sub.8 is used as an oxidizing agent of
Ag(I) ions (see the discussion below relating to oxidation with K.sub.
2S.sub.2O.sub.8).

FIG. 2 provides a SEM micrograph of the grayish black precipitate. The
smaller "cubical" particles represent Ag.sub.7O.sub.8NO.sub.3 and
their size, based on SEM is estimated at about 2.5 .mu.m. The shape of
these particles was found to be in very good agreement with the
results of Skanavi-Grigoreva (M. S. Skanavi-Grigoreva, I. L.
Shimanovich, Zh. Obsh., Khim., 24, 1490(1954)). Who produced this
material by the electrolysis of an aqueous AgNO.sub.3 solution. The
larger, cylindrical particles represent silver sulfate (Ag.sub.2SO.sub.
4).

(c) Deposition of Silver Deposition Products Using K.sub.2S.sub.2O.sub.
8

Some differences in the formation of the grayish-black precipitate
were observed when K.sub.2S.sub.2O.sub.8 was used instead of (NH.sub.
4).sub.2S.sub.2O.sub.8, as the oxidizing agent of Ag(I). The
precipitation of the grayish-black compound was significantly faster,
and occurred within 1 minute upon addition of K.sub.2S.sub.2O.sub.8 to
the AgNO.sub.3 solution. During this time, the pH of the solution
changed from the initial pH of about 7 to below 1 after the
precipitation.

(d) Properties of Deposition Products Produced Using K.sub.2S.sub.
2O.sub.8

As determined by XRD analysis in FIG. 3, all the peaks in the pattern
exactly match the compound of composition Ag.sub.7O.sub.8NO.sub.3. No
other compounds were identified in this XRD pattern.

The theoretical amount of Ag in the compound Ag.sub.7O.sub.8NO.sub.3
is 79.90%. The chemical analysis determined that the grayish black
precipitate contained about 78.80% Ag. This result shows a good
agreement of the experiments with the theory.

The SEM micrographs of the powder produced by the chemical oxidation
of AgNO.sub.3 with K.sub.2S.sub.2O.sub.8 are presented in FIG. 4. It
appears that the particles are uniform and cubical in their shape. The
size of these particles is estimated at about 2.5 .mu.m.

(e) Antimicrobial Activity

The comparison of the SEM micrographs of uncoated and coated HDPE mesh
samples is presented in FIG. 5. As shown in FIG. 5, the surface of the
HDPE is partially covered with the Ag(Ag.sub.3O.sub.4).sub.2NO.sub.3
particulates.

These samples were tested for bioactivity against the bacteria
Pseudomonas aeruginosa, Staphylococcus aureus or fungi Candida
albicans. As can be seen from the photographs presented in FIG. 6,
clear zones surrounding the test samples (where a growth of tested
microorganisms did not occur) were observed in all cases for
Staphylococcus aureus (a gram-positive bacteria), Pseudomonas
aeuguginosa (a gram-negative bacteria) and Candida albicans (an
example of fungi). The size of the controlled zone of inhibition
(CZOI), where the growth of tested microorganisms was not observed,
was estimated at 3 mm to 5 mm for all tested samples. These results
suggest that the deposition products have antibacterial and antifungal
properties. Furthermore these results are in agreement with previously
published results, where was suggested that only oxidized silver
species, but not metallic silver exhibit an antimicrobial activity.

(f) Conclusions Relating to Examples 15-16

It has been demonstrated that deposition products, namely those of
composition Ag.sub.7NO.sub.11.times.3Ag.sub.2SO.sub.4 or Ag.sub.
7NO.sub.11 can successfully be deposited as powders or on a substrate
such as HDPE mesh, by a simple reaction between AgNO.sub.3 and (NH.sub.
4)S.sub.2O.sub.8 or K.sub.2S.sub.2O.sub.8. These compounds are soluble
in both concentrated HNO.sub.3 or NH.sub.4OH.

Example 17

Samples of a substrate consisting of a patterned wound dressing made
of a perforated plastic carrier material with a skin adhesive layer
comprised of a hydrophobic cross-linked silicon gel (trade-mark
Mepitel.TM., product of Molnlycke Health Care AB, Sweden) were
subjected to SEM micrography to observe the density and coverage on
the substrate of a deposition product deposited on the substrate in
accordance with the second and third aspects of the invention, and to
XRD analysis to analyze the composition of the deposition product
deposited on the substrate.

FIG. 7 depicts an uncoated sample of the Mepitel.TM. wound dressing at
a magnification of 30.times.. FIGS. 8-11 depict samples of composite
materials which have been produced according to the second and third
aspects of the invention in the same manner as described in Example
14.

FIG. 8 depicts a composite material comprising a coated sample of the
Mepitel.TM. wound dressing at a magnification of 40.times., in which a
relatively low amount of deposition product has been deposited on the
substrate. FIG. 9 depicts the composite material of FIG. 8 at a
magnification of 2000.times., and clearly shows that the density and
coverage of the deposition product is such that the skin adhesive
layer of the Mepitel.TM. wound dressing is relatively unobstructed by
the deposition product.

FIG. 10 depicts a composite material comprising a coated sample of the
Mepitel.TM. wound dressing at a magnification of 40.times., in which a
higher amount of deposition product has been deposited on the
substrate in comparison with FIG. 8 and FIG. 9. FIG. 11 depicts the
composite material of FIG. 10 at a magnification of 2000.times., and
clearly shows that the skin adhesive layer remains relatively
unobstructed by the deposition product.

FIG. 12 depicts an XRD pattern for an uncoated sample of the
Mepitel.TM. wound dressing. FIG. 13 depicts an XRD pattern for a
composite material comprising a sample of the Mepitel.TM. wound
dressing which has been coated with a deposition product according to
the second and third aspects of the invention in the same manner as
described in Example 14. FIG. 14 superimposes the XRD patterns from
FIG. 12 and FIG. 13.

Referring to FIG. 14, the peaks which are observed in the pattern from
FIG. 13 but which are not observed in the pattern from FIG. 12 may be
attributed to the deposition product. These peaks define the
deposition product as comprising at least some amount of Ag.sub.7O.sub.
8NO.sub.3.

Example 18

Samples of a substrate consisting of a patterned wound dressing made
of a perforated plastic carrier material with a skin adhesive layer
comprised of a hydrophobic cross-linked silicon gel (trade-mark
Mepitel.TM., product of Molnlycke Health Care AB, Sweden) coated with
0.6 mg/cm2 of total silver according to the second and third aspects
of the invention in the same manner as described in Example 14 were
exposed to a solution containing 10 g/L Na.sub.2S. After 10 minutes of
exposure to the Na.sub.2S solution the coated wound dressing samples
were carefully washed with water until pH 7.

After drying, the samples were tested for antimicrobial activity
against Pseudomonas aeruginosa and Staphylococcus aureus using
standard procedures. Clear zones of inhibition of bacterial growth
surrounding test samples were observed for both Pseudomonas aeruginosa
and Staphylococcus aureus, suggesting that a deposition product
produced according to the second and third aspects of the invention
will exhibit an antimicrobial activity even after exposure to a
sulfide containing environment.



United States Patent 7,300,673
Djokic November 27, 2007
Deposition products, composite materials and processes for the
production thereof

Abstract

Methods for the production of deposition products including an
oxidized metal species under both acidic and alkaline conditions,
methods for the production of composite materials including a
substrate and the deposition product, and products in the nature of
deposition products and composite materials. The methods are
particularly suited for depositing a very thin layer of a deposition
product on a substrate and may be used to produce composite materials
for use in many applications, including but not limited to
electronics, materials engineering and medical applications. In a
preferred embodiment, the metal is silver, the substrates are medical
devices or components of medical devices and the deposition product
includes an antimicrobially active oxidized silver species.
Inventors: Djokic; Stojan (Edmonton, CA)
Assignee: Exciton Technologies Inc. (Edmonton, CA)
Appl. No.: 10/830,574
Filed: April 23, 2004
Foreign Application Priority Data

May 16, 2003 [CA] 2428922
Mar 10, 2004 [CA] 2460585

Current U.S. Class: 424/618 ; 424/443; 424/445; 424/613; 424/703;
514/495; 514/709
Current International Class: A61K 33/38 (20060101); A61K 31/10
(20060101); A61K 31/28 (20060101); A61K 33/04 (20060101); A61K 33/40
(20060101); A61K 9/70 (20060101); A61L 15/00 (20060101); A01N 39/00
(20060101); A01N 41/10 (20060101); A01N 55/02 (20060101); A01N 59/02
(20060101); A01N 59/16 (20060101)
Field of Search: 514/495,709 424/613,618,443,445,703
References Cited [Referenced By]
U.S. Patent Documents

3619387 November 1971 Mindt et al.
3998602 December 1976 Horowitz et al.
4411981 October 1983 Minezaki
4439557 March 1984 Kawamura et al.
4728323 March 1988 Matson
5019096 May 1991 Fox et al.
5098582 March 1992 Antelman
5207873 May 1993 Sanduja et al.
5211855 May 1993 Antelman
5372847 December 1994 Silver et al.
5413788 May 1995 Edwards et al.
5474797 December 1995 Sioshansi et al.
5676977 October 1997 Antelman
5681575 October 1997 Burrell et al.
5814094 September 1998 Becker et al.
5928174 July 1999 Gibbins
5985308 November 1999 Burrell et al.
6017553 January 2000 Burrell et al.
6080490 June 2000 Burrell et al.
6087549 July 2000 Flick
6224983 May 2001 Sodervall
6238686 May 2001 Burrell et al.
6267782 July 2001 Ogle et al.
6306341 October 2001 Yokota et al.
6333093 December 2001 Burrell et al.
6355858 March 2002 Gibbins
6379712 April 2002 Yan et al.
6426195 July 2002 Zhong et al.
6436420 August 2002 Antelman
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2002/0127282 September 2002 Antelman
Foreign Patent Documents

2102433 Mar., 2000 CA
1149389 May., 1997 CN
0 875 146 Nov., 1998 EP
431656 Jul., 1935 GB
WO 2004/037187 May., 2004 WO

Other References

Djokic, S. Journal of the Electrochemical Society 2004, 151(6), C359-
C364. cited by examiner .
N.R. Thompson, Silver, in Comprehensive Inorganic Chemistry, v. IIID,
J. C. Bailer et al. , Editors, Pergamon Press, Oxford (1973) 79-80.
cited by other .
K.J. Bundy et al., "An Investigation of the Bacteriostatic Properties
of Pure Metals," Journal of Biomedical Materials Research, v. 14
(1980) 653-662. cited by other .
D. Acel, "Uber die oligodynamische Wirkung der Metalle," Z. Biochem,
112 (1920) 23-26, with English abstract. cited by other .
J. Gibbard, "Public Health Aspects of the Treatment of Water and
Beverages with Silver," Journal of American Public Health, v. 27
(1937) 112-119. cited by other .
S.S. Djokic and R.E. Burrell, "Behavior of Silver in Physiological
Solutions," Journal of Electrochemical Society. v. 145(5) (1998)
1426-1430. cited by other .
S.S. Djokic et al., "An Electrochemical Analysis of Thin Silver
Produced by Reactive Sputtering," J. of Electrochemical Society, v.
148(3) (2001) C191-C196. cited by other .
C.L. Fox, "Topical Therapy and the Development of Silver
Sulfadiazine," Surgery, Gynecology & Obstetrics, 157 (1983) 82-88.
cited by other .
M.C. Fung, D.L. Bowen, "Silver Products for Medical Indications: Risk-
Benefit Assessment," Clinical Toxicology, v. 34(1) (1996) 119-126.
cited by other .
H.W. Margaf, T.H. Covey, "A Trial of Silver-Zinc-Allantoinate in the
Treatment of Leg Ulcers," Arch. Surg., v. 112 (1977) 699-704. cited by
other .
S.I. Zhdanov, "Sulfur, Selenium, Tellurium and Polonium," in Standard
Potentials in Aqueous Solutions, A.J. Bard et al. Editors, Marcel
Dekker Inc., New York (1985) 93, 5 pp. cited by other .
M.S. Skanavi-Grigoreva, I.L. Shimanovich, Zh. Obsh., Khim., 24, (1954)
1490-1495, with English abstract. cited by other .
"Electrocrystallization," article from website: www.mpi-stuttgart.mpg.de/jansen/p110,
2 pages. cited by other .
Leibecki, Harold F., "Argentic Oxysalt Electrodes," NASA Technical
Note NASA TN D-3208, Washington, D.C., Jan. 1966. cited by other .
McMillan, J.A.,"Higher Oxidation States of Silver," Chem. Rev.
(Washington, D.C.), 62, 1962, pp. 65-80. cited by other .
Discussion Section, Journal of The Electrochemical Society, vol. 106,
No. 12, Dec. 1959, pp. 1072-1084. cited by other.

Primary Examiner: Richter; Johann R.
Assistant Examiner: Arnold; Ernst V
Attorney, Agent or Firm: Kuharchuk; Terrence N. Rodman & Rodman
Claims


The embodiments of the invention in which an exclusive property or
privilege is claimed are defined as follows:

1. A method for producing a composite material comprising a substrate
and a deposition product, the method comprising the following steps:
(a) providing a deposition solution comprising an amount of an aqueous
solution of a silver salt comprised of an amount of silver ions and an
amount of an oxidizing agent, wherein the silver salt is comprised of
silver nitrate, wherein the oxidizing agent is comprised of a
persulfate, wherein a concentration of the silver salt in the
deposition solution is between about 1 gram per liter and about 20
grams per liter, wherein a concentration of the oxidizing agent in the
deposition solution is between about 1 gram per liter and about 50
grams per liter, and wherein the deposition solution has a pH below 7;
(b) producing the deposition product by facilitating a chemical
reaction in the deposition solution between the silver ions and the
oxidizing agent while maintaining the deposition solution at a
temperature of between about 2 degrees Celsius and about 40 degrees
Celsius, wherein the deposition product consists essentially of at
least one oxidized silver species, and wherein the deposition product
is comprised of a compound having the formula Ag.sub.7O.sub.8X, where
X is an anion; (c) providing the substrate, wherein the substrate is a
medical device; and (d) contacting the substrate with the deposition
solution during the deposition product producing step (b), thereby
producing the composite material.

2. The method as claimed in claim 1 wherein the persulfate is selected
from the group of persulfates consisting of potassium persulfate,
sodium persulfate, ammonium persulfate and mixtures thereof.

3. The method as claimed in claim 2 wherein the persulfate is
comprised of potassium persulfate.

4. The method as claimed in claim 1 wherein the amount of the
oxidizing agent is selected to be a stoichiometrically appropriate
amount relative to the amount of the silver ions.

5. The method as claimed in claim 2, further comprising the step of
adding an amount of a source of anions to the deposition solution for
combining with the silver ions in order to produce the deposition
product.

6. The method as claimed in claim 5 wherein the source of anions is
comprised of at least one acid.

7. The method as claimed in claim 6 wherein the acid is selected from
the group of acids consisting of carbonic acid, nitric acid,
perchloric acid, sulfuric acid, acetic acid, fluoroboric acid, citric
acid, acetylsalicylic acid and mixtures thereof.

8. The method as claimed in claim 5 wherein the amount of the source
of anions is selected to be a stoichiometrically appropriate amount
relative to the amount of the silver ions.

9. The method as claimed in claim 2 wherein the deposition product
producing step is comprised of maintaining the deposition solution at
a temperature of between about 10 degrees Celsius and about 25 degrees
Celsius.

10. The method as claimed in claim 2 wherein the deposition product
producing step is comprised of agitating the deposition solution
during at least a portion of the deposition product producing step.

11. The method as claimed in claim 1 wherein the substrate contacting
step is performed for at least about 1 minute.

12. The method as claimed in claim 11 wherein die substrate contacting
step is performed for between about 1 minute and about 60 minutes.

13. The method as claimed in claim 12 wherein the substrate contacting
step is performed for between about 1 minute and about 20 minutes.

14. The method as claimed in claim 13 wherein the substrate contacting
step is performed for between about 2 minutes and about 10 minutes.

15. The method as claimed in claim 1, further comprising the step,
following the substrate contacting step, of washing the composite
material.

16. The method as claimed in claim 1, further comprising the step,
prior to the substrate contacting step, of etching the substrate by
immersing the substrate in an etching solution.

17. The method as claimed in claim 16 wherein the etching solution is
comprised of a mixture of an alcohol and an aqueous solution of a
hydroxide compound.

18. The method as claimed in claim 17 wherein the hydroxide compound
is selected from the group of hydroxide compounds consisting of sodium
hydroxide, potassium hydroxide and mixtures thereof.

19. The method as claimed in claim 17 wherein the hydroxide compound
is comprised of sodium hydroxide.

20. The method as claimed in claim 17 wherein the etching step is
performed for between about 5 minutes and about 20 minutes.

21. The method as claimed in claim 17, further comprising the step,
following the etching step, of washing the substrate to remove
residual alkali from the substrate.

22. The method as claimed in claim 1, further comprising the step,
following the substrate contacting step, of immersing the composite
material in boiling water.

23. The method as claimed in claim 22 wherein the immersing step is
performed for at least about 1 minute.

24. The method as claimed in claim 1 wherein the substrate is
comprised of a wound dressing.

25. The method as claimed in claim 24 wherein the substrate is
comprised of a high density polyethylene material.

26. The method as claimed in claim 25 wherein the substrate is
comprised of a skin adhesive layer, wherein the skin adhesive layer is
comprised of a cross-linked silicon gel, and wherein the deposition
product is deposited on the skin adhesive layer.

27. The method as claimed in claim 26 wherein the skin adhesive layer
has adhesive properties and wherein the deposition product does not
materially interfere with the adhesive properties of the skin adhesive
layer.

28. The method as claimed in claim 1 wherein the deposition product is
further comprised of Ag.sub.2SO.sub.4.

29. The method as claimed in claim 1 wherein the deposition product is
further comprised of at least one silver oxide selected from the group
of silver oxides consisting of monovalent silver oxide, bivalent
silver oxide, trivalent silver oxide and mixtures thereof.

30. The method as claimed in claim 1 wherein X is derived from an
acid.

31. The method as claimed in claim 1 wherein the deposition product is
comprised of a plurality of valent states of silver.

32. The method as claimed in claim 1 wherein X is selected from the
group of anions consisting of HCO.sub.3.sup.-, CO.sub.3.sup.2-, NO.sub.
3.sup.-, ClO.sub.4.sup.-, SO.sub.4.sup.2-, F.sup.-, and mixtures
thereof.

33. The method as claimed in claim 32 wherein X is comprised of NO.sub.
3.sup.-.
Description


FIELD OF INVENTION

Deposition products, composite materials including deposition
products, and methods for producing the deposition products and the
composite materials.

BACKGROUND ART

The germicidal properties of silver, even not known as such, have been
utilized since the early Mediterranean cultures. It has been known
since 1000 BC and possibly before that water kept in silver vessels
and then exposed to light and filtered could be rendered potable.
Other forms of silver have been used throughout centuries for various
applications, such as coatings for prevention of beverages from
spoilage or silver plates and foils in the surgical treatments of
wounds and broken bones.

The lethal effects of metals towards bacteria and lower life forms
were first scientifically described by von Nageli in the late
nineteenth century, and this phenomenon has been defined as an
"oligodynamic effect" (N. R. Thompson, Silver, in Comprehensive
Inorganic Chemistry, Vol. III D, J. C. Bailer, H. J. Emeleus, R.
Nyholm and A. F. Trutman-Dickenson, Editors, Pergamon Press, Oxford
(1973)). The term oligodynamic effect is typically restricted to
describing solutions in which the metal concentration is several
orders of magnitude lower than that which would be lethal to higher
organisms.

The investigation of the bacteriostatic properties of pure metals such
as Fe, Mo, Cu, V, Sn, W, Au, Al, Ta, Nb, Ti, Zr, Ni, Co, Ag and Cr,
has proved that Co was the only element which was inhibitory for the
bacterial growth under anaerobic conditions (K. J. Bundy, M. F. Butler
and R. F. Hochman, "An Investigation of the Bacteriostatic Properties
of Pure Metals", Journal of Biomedical Materials Research, Vol. 14
(1980) 653-663). Under aerobic conditions both Cu and Co consistently
display inhibitory effects. Some antimicrobial effects have been seen
for Ni, Fe and V. However, other metals such as Mo, W, Al, Nb, Zr, Cr
and most importantly for the present invention Ag and Sn never showed
any tendency to inhibit the growth of Streptococcus mutans.

In the case of silver metal, it was in 1920, when Acel who was the
first to attribute the antimicrobial properties of silver to the
liberation of Ag.sup.+ ions from the material (D. Acel, "Uber die
oligodynamische Wirkung der Metalle", Z. Biochem., 112 (1920) 23).

Gibbard reported in 1937 that pure metallic silver has no
antimicrobial activity (J. Gibbard, "Public Health Aspects of the
Treatment of Water and Beverages with Silver", Journal of American
Public Health, Vol. 27 (1937) 112-119). His experiments showed that if
silver is cleaned mechanically with an abrasive cloth or paper it
becomes inactive. Similarly, if molten silver is allowed to cool in a
reduction atmosphere (e.g. hydrogen), no antimicrobial activity is
found. When cooling of molten silver is carried out in air, and
formation of surface oxide occurred, an antimicrobial activity may be
observed. Similar results were found when silver metal was treated
with nitric acid in an air atmosphere (dissolution and formation of an
oxide layer). Based on Gibbard's results, pure silver was devoid of
activity, but surface oxidized silver was active. Silver oxide, silver
nitrate and silver chloride were always active. Also, Gibbard observed
that the antimicrobial properties of silver and its compounds were
reduced in the presence of proteins or glucose.

Djoki investigated the behavior of silver films, e.g. physical vapor
deposited, electrodeposited, electroless deposited and metallurgical
in physiological saline solutions (S. S. Djoki and R. E. Burrell,
"Behavior of Silver in Physiological Solutions", Journal of the
Electrochemical Society, Vol. 145 (5) (1998) 1426-1430). Djoki found
that an essential factor leading to an antimicrobial activity of
metallic silver is a presence of Ag oxide(s) at the surface of this
material. It was demonstrated that only silver films containing silver
oxides (most likely Ag.sub.2O) showed an antimicrobial activity. The
behavior was attributed to the dissolution of Ag.sub.2O from the
"silver" material and formation of Ag.sup.+ or other complexed ions
which become antimicrobially active. There was no evidence that pure
metallic silver, no matter which way it was produced i.e., physical
vapor deposited, electrodeposited or electroless deposited could be
dissolved in physiological media, or that these materials would
exhibit antimicrobial activity.

It should be noted that when the physical vapor deposition of silver
was carried out in an atmosphere containing oxygen the resulted
product, as found by the XRD analysis contained silver oxide.
Consequently, these samples exhibited antimicrobial activity.
Conversely, when the physical vapor deposition was carried out from an
argon atmosphere (no presence of oxygen) pure metallic,
nanocrystalline silver film was deposited as confirmed by the XRD
analysis. However, these films did not dissolve in physiological
saline solutions, nor they exhibited antimicrobial activity at all.

For an in depth understanding of structural properties of silver films
produced by reactive sputtering, see Djoki et al. (S. S. Djoki , R. E.
Burrell, N. Le and D. J. Field, "An Electrochemical Analysis of Thin
Silver Produced by Reactive Sputtering", Journal of the
Electrochemical Society, Vol. 148 (3) (2001) C191-C196.). To prove the
concept that only oxidized silver species are responsible for the
antimicrobial activity, Djoki further oxidized pure metallic silver
samples (i.e. those produced by the electrodeposition, electroless
deposition, physical vapor deposition in an argon atmosphere or
metallurgically). The oxidation of these samples was carried out
electrochemically in 1 M KOH solutions, using a process very well
established in the art. The electrochemically oxidized silver samples
were tested for the antimicrobial activity against Pseudomonas
Aeruginosa. Clear evidence was found that the electrochemically
oxidized silver samples exhibited antimicrobial activity.

The above referenced work shows that only oxidized silver species, but
not elemental silver will affect antimicrobial activity. The findings
to date show that the "nanocrystalline" or "macrocrystalline"
elemental silver does not have antimicrobial activity at all.
Elemental silver, either nanocrystalline or "macrocrystalline" may
exhibit some antimicrobial activity only if oxidized silver species
are present at these surfaces or within the silver metal. Only the
formation of silver oxide(s), carbonates or other silver salts (except
silver sulfide, due to its extremely low solubility) at the surface or
within the material, which may be influenced by an exposure of
elemental silver to various bases, acids or due to atmospheric
corrosion may lead to an antimicrobial activity of this material.

The use of silver on chronic wounds dates back in the 17.sup.th and
18.sup.th centuries. In the early 19.sup.th century, silver nitrate
began to be used on burns and in ophthalmology. Concentrations of the
solution ranged from 0.20 to 2.5 wt. % with the weaker solutions being
reserved for children. Silver has been found to be active against a
wide range of bacterial, fungal and viral pathogens. Topical treatment
of acute and chronic wounds is a preferred and selective approach to
the prevention of infection and healing. In order to achieve these
requirements products that are used in the prevention of infections
must have certain physical and chemical properties.

When used for topical dressings, silver compounds must have relatively
low solubility. This is usually achieved by choosing compounds with a
relatively low solubility products (e.g. AgCl, Ag.sub.2SO.sub.4,
Ag.sub.2CO.sub.3, Ag.sub.3PO.sub.4, Ag-oxides). Kinetics of
dissolution of these compounds in neutral aqueous solutions is quite
slow. This property is very convenient for two reasons. First, a
sustained release of silver ions from the silver compounds is more
likely to provide a prolonged antimicrobial activity. Second, low
amounts of the silver ions released into wound exudates may not give
rise to transient high tissue blood and urine levels, thus avoiding
systemic toxicity. The choice of a particular silver compound will
depend upon its reactivity with wound exudates. This reactivity should
preferably be minimized in order to achieve the desired effect of the
released silver ions (i.e., antimicrobial activity without systemic
toxicity).

Besides silver nitrate, one of the most widely used topical
antimicrobial materials is silver sulfadiazine (C. L. Fox, "Topical
Therapy and the development of Silver Sulfadiazine", Surgical
Gynecology & Obstetrics, 157 (1) (1983) 82-88). This compound is
synthesized from silver nitrate and sodium sulfadiazine. Silver
sulfadiazine has been used in treatments of burns, leg ulcers and also
as a topical antimicrobial agent in the management of infected wounds.

Products such as silver protein (argyrols) or mild silver protein are
mixtures of silver nitrate, sodium hydroxide and gelatin. These
products are recommended for internal use and are promoted as
essential mineral supplements. Although there is no theoretical or
practical justification for their use, this class of compounds has
been recommended for the treatment of diverse diseases such as cancer,
diabetes, AIDS and herpes (M. C. Fung, D. L. Bowen, "Silver Products
for Medical Indications: Risk--Benefit Assessment", Clinical
Toxicology, Vol. 34 (1) (1996) 119-126).

Silver-zinc-allantoinate has been formulated as a cream and represents
a combination of silver, zinc and allantoin (an agent that stimulates
debridement and tissue growth (H. W. Margaf, T. H. Covey, "A Trial of
Silver-Zinc-Allantoinate in the Treatment of Leg Ulcers", Arch. Surg.,
Vol. 112 (1977) 699-704). This composition exhibited promising effects
in preliminary studies.

In the past few decades several topical dressings containing silver
have been developed for wound care. Such materials include
Arglaes.TM., Silverlon.TM., Acticoat.TM., Actisorb.TM., and Silver
220.TM..

Antimicrobial coatings and methods of forming same are the subject of
U.S. Pat. No. 5,681,575 (Burrell et al) and U.S. Pat. No. 6,238,686
(Burrell et al). The coatings are formed by the physical vapour
deposition of biocompatible metal and the preferred biocompatible
metal is silver.

Burrell et al teach that atomic disorder may be created in metal
powders or foils by cold working and in metal coatings by depositing
by vapor deposition at low substrate temperatures and that such metal
coatings constitute a matrix containing atoms or molecules of a
different material. The presence of different atoms or molecules
results in atomic disorder in the metal matrix, for instance due to
different sized atoms. The different atoms or molecules may be one or
more second metals, metal alloys or metal compounds which are co- or
sequentially deposited with the first metal or metals to be released.
Alternatively, the different atoms or molecules may be adsorbed or
trapped from the working gas atmosphere during reactive vapor
deposition.

In U.S. Pat. No. 6,238,686 Burrell et al claim a modified material
comprising one or more metals in a form characterized by sufficient
atomic disorder such that the material, in contact with a solvent for
the material, releases atoms, ions, molecules or clusters containing
at least one metal at an enhanced rate relative to its normal ordered
crystalline structure. In U.S. Pat. No. 5,681,575 Burrell et al claim
a medical device which includes a coating of one or more anti-
microbial metals having a "sufficient atomic disorder".

It is unclear from either U.S. Pat. No. 5,681,575 or U.S. Pat. No.
6,238,686 what would constitute a material characterized by
"sufficient atomic disorder". In nature, most materials would exhibit
sufficient atomic disorder if the true atomic disorder described (by
drawings or mapping) in ordinary Chemistry or Physics handbooks were
insufficiently ordered (with a regular geometric structure or like).

In any event, the teachings of Burrell et al appear to connect "atomic
disorder" with an "enhanced rate" of release of "atoms, ions,
molecules or clusters". If the term "release" further relates to a
dissolution (as defined in textbooks of General Chemistry and
Physics), then this dissolution should lead to the liberation of ions
or molecules in solvent. When released in the solvent, these ions or
molecules are usually solvated i.e. surrounded by the molecules of the
solvent. It is very unlikely that atoms of a metal will be released
into a solution comprising of water such as in the wound environment.
If released into solution in its elemental state, metals would rather
represent a relatively larger particles comprising of more than one or
a few atoms.

As a result, the term "atom" as used in Burrell et al is not exactly
descriptive. It is not known yet scientifically whether atoms of
metals can be released into aqueous solutions at pH close to neutral
(e.g., pH range 6 to 8), except in the case of colloidal solutions
which are usually prepared by adequate chemical reactions in-situ.

U.S. Pat. No. 6,087,549 (Flick) discloses a multilayer laminate wound
dressing comprising a plurality of layers of a fibrous material, with
each layer comprising a unique ratio of metalized fibers to
nonmetalized fibers. In a preferred embodiment the wound dressing
consists of three layers and the metal is silver. The wound contact
layer has the highest ratio of metalized fibers to nonmetalized
fibers, the intermediate layer has a lower ratio of metalized fibers
to nonmetalized fibers, and the outer layer has the lowest ratio of
metalized fibers to nonmetalized fibers. The wound dressing described
by Flick is commercially available under the trade-mark Silverlon.TM..

U.S. Pat. No. 5,211,855 (Antelman), U.S. Pat. No. 5,676,977 (Antelman)
and U.S. Pat. No. 6,436,420 (Antelman) teach that tetrasilver
tetroxide (Ag.sub.4O.sub.4) containing two monovalent and two
trivalent silver ions exhibits bactericidal, fungicidal and algicidal
properties. As a result, "tetrasilver tetroxide" is suggested for use
for water treatment in U.S. Pat. No. 5,211,855 and for use in
destroying the AIDS virus in U.S. Pat. No. 5,676,977.

In U.S. Pat. No. 6,436,420, Antelman describes a method of deposition
or interstitial precipitation of tetrasilver tetroxide (Ag.sub.4O.sub.
4) crystals within the interstices of fibers, yams and/or fabrics
forming such articles in order to produce fibrous textile articles
possessing enhanced antimicrobial properties. The interstitial
precipitation of Ag.sub.4O.sub.4 is achieved by immersion of the
article to be treated (e.g., fiber, yam or fabric) in an aqueous
solution containing a water soluble silver salt, most preferably
silver nitrate. After uniformly wetting the article, the article is
removed into a second heated aqueous solution (having a temperature of
at least 85 degrees Celsius or more preferably at least 90 degrees
Celsius) containing strong alkali (most preferably NaOH) and a water
soluble oxidizing agent (most preferably potassium persulfate) for 30
seconds to 5 minutes to facilitate the precipitation of tetrasilver
tetroxide.

After the reaction is completed, the article is removed and washed.
The article treated in this way is described as exhibiting outstanding
antimicrobial resistance towards pathogens such as bacteria, viruses,
yeast and algae. The article is also described as being resistant to
ultraviolet light and as maintaining its antimicrobial properties
after a number of launderings.

SUMMARY OF INVENTION

The present invention is directed at deposition products, composite
materials and at methods for the production of deposition products and
composite materials. The deposition products are comprised of at least
one oxidized species of a metal.

The methods of the invention are based upon chemical deposition
principles and techniques. The methods of the invention may be carried
out under either acidic or alkaline conditions. The methods of the
invention may comprise the step of exposing ions of the metal to an
oxidizing agent to produce the deposition product. The methods of the
invention may involve the production of the deposition product itself
or the production of a composite material which comprises a substrate
and the deposition product.

The methods of the invention are particularly suited for producing a
composite material which is comprised of a substrate and a very thin
coating or deposition layer of the deposition product. This thin
coating or layer may be in the order of one or several atoms in
thickness, which facilitates the production of a composite material
which has a relatively high surface area to volume ratio. The coating
may also be deposited so that it does not completely cover the
substrate, thus leaving portions of the surface of the substrate
uncoated. Composite materials produced using the methods of the
invention may be useful for a variety of applications, including but
not limited to electronics, materials engineering and medical
applications.

The methods of the invention may be carried out at relatively low
temperatures. Preferably the methods of the invention are carried out
at temperatures of no greater than about 60 degrees Celsius. More
preferably the methods of the invention are carried out at room
temperature (i.e., between about 10 degrees Celsius and about 25
degrees Celsius).

The metal and the oxidizing agent are selected so that they are
compatible with the production of the desired deposition product. As a
result, any suitable metal and any suitable oxidizing agent may be
used in the invention. The metal may also be comprised of more than
one element, with the result that the deposition product may be
comprised of at least one oxidized species of more than one metal
element.

Preferably the metal is comprised of silver and the deposition product
is comprised of at least one oxidized species comprising silver. The
metal may, however, be further comprised of other metal elements such
as gold, copper, tin or zinc so that the deposition product is
comprised of at least one oxidized species comprising silver and one
or more other metals.

Where the metal is comprised of silver, the resulting deposition
product may exhibit significant antimicrobial properties. Without
intending to be limited by theory, it is believed that these
antimicrobial properties are due to the presence in the deposition
product of one or more oxidized silver species. The presence of other
metals in the deposition product may enhance these antimicrobial
properties or may provide other complementary properties to the
deposition product.

More particularly, it is believed that silver containing deposition
products produced using the methods of the invention may be comprised
of silver ions having valent states higher than one, such as for
example Ag (II) and Ag (III) valent states. It is also believed that
silver containing deposition products produced using the methods of
the invention may be comprised of silver ions having more than one
valent state so that the oxidized silver species may be comprised of a
multivalent substance. Finally, it is believed that silver containing
deposition products produced using the methods of the invention may be
comprised of a silver containing substance or a plurality of silver
containing substances which react over time to form other silver
containing substances which may exhibit differing antimicrobial
properties. It is believed that if this is the case, the deposition
products produced by the invention may be useful for providing a
varied antimicrobial response and for overcoming bacterial resistance.

In particular, in certain aspects, the methods of the invention may be
used to produce a deposition product which comprises a substance
having the general formula Ag.sub.7O.sub.8X, where X is an anion. The
deposition product may be further comprised of Ag.sub.2SO.sub.4. The
deposition product may also be comprised of other oxidized silver
compounds such as one or more silver oxides selected from the group of
silver oxides consisting of monovalent silver oxide, bivalent silver
oxide, trivalent silver oxide and mixtures thereof.

The anion X may be comprised of a single anion or may be comprised of
a plurality of different anions. The anion may therefore be comprised
of any anion or combination of ions. The anion may, for example, be
selected from the group of anions consisting of HCO.sub.3.sup.-,
CO.sub.3.sup.2-, NO.sub.3.sup.-, ClO.sub.4.sup.-, SO.sub.4.sup.2-,
F.sup.-, and mixtures thereof. The source of the anion may be a metal
compound which provides the ions of the metal. For example, where the
deposition solution is comprised of a silver salt such as silver
nitrate, the anion may be comprised of the nitrate ion (NO.sub.
3.sup.-). An alternative or secondary source of the anion X may
optionally be provided in order to provide sufficient quantities of
the anion for production of the deposition product. Where an
alternative or secondary source of the anion X is provided, the source
of anions may be comprised of any source, including but not limited to
any organic or inorganic acid.

Where the metal is comprised of silver, the composite materials
produced by the methods may therefore be useful as medical devices or
as components of medical devices due to their specific antimicrobial
properties. These composite materials may also provide other
advantages. As one example, the ability to provide a very thin coating
or layer of the deposition product on the substrate makes it possible
to minimize the amount of silver which must be used in the composite
material in order to provide a desired antimicrobial response. As a
second example, the ability to provide a very thin coating or layer of
the deposition product on the substrate minimizes the extent to which
the deposition product will interfere with the properties and
functions of the substrate, particularly if the deposition product is
deposited on the substrate so that it does not completely cover the
surface of the substrate. This second example may be particularly
significant where the substrate is comprised of an adhesive material
such as a skin adhesive layer.

In a first aspect, the invention is a method for producing a composite
material comprising a substrate and a deposition product, wherein the
deposition product is comprised of at least one oxidized species of a
metal, the method comprising the following steps: (a) first contacting
the substrate with a first basic environment comprising ions of the
metal in order to expose the substrate to the ions of the metal; and
(b) second contacting the substrate with a second basic environment in
order to produce the composite material.

The first basic environment may be comprised of any environment in
which metal ions are present under alkali conditions. The metal may be
comprised of any metal or combinations of metals but preferably the
metal is comprised of silver.

Preferably the first basic environment is comprised of a first basic
solution comprising an amount of a silver diamino complex. More
preferably, the first basic solution results from a mixture of a
silver compound and ammonium hydroxide in an aqueous medium.
Preferably the silver compound is selected from the group of silver
compounds consisting of silver salts, silver oxides and mixtures
thereof. More preferably the silver compound is comprised of silver
nitrate.

The first basic solution may have any alkaline pH. Preferably the
first basic solution has a pH in the range from about 8 to about 14.
Within these parameters, the amount of ammonium hydroxide in the first
basic solution is preferably selected such that a concentration of
ammonium hydroxide in the first basic solution is between about 25
percent and about 35 percent by volume of the first basic solution.
Preferably the amount of silver compound in the first basic solution
is selected such that a concentration of the silver compound in the
first basic solution is between about 1 gram per liter and about 20
grams per liter.

The second basic environment may be comprised of any environment
having alkali conditions. Preferably the second basic environment is a
strongly alkaline environment having a pH at least about 12.
Preferably the second basic environment is comprised of a second basic
solution containing an amount of a strong alkali compound. The strong
alkali compound may be comprised of any compound which can provide the
strong alkaline environment. For example, the strong alkali compound
may be comprised of one or more Group I elements, including lithium,
sodium, potassium, rubidium, cesium and francium. Preferably the
strong alkali compound is selected from the group of compounds
consisting of sodium hydroxide and potassium hydroxide and mixtures
thereof and more preferably the strong alkali compound is comprised of
sodium hydroxide. Preferably the amount of hydroxide compound in the
second basic solution is selected such that a concentration of the
hydroxide compound in the second basic solution is between about 15
grams per liter and about 35 grams per liter.

The first contacting step may be performed for any length of time
which is sufficient to expose the substrate to the ions of the metal.
Preferably the substrate is substantially completely exposed to the
ions of the metal. Preferably the first contacting step is performed
for between about 1 minute and about 10 minutes.

The second contacting step may be performed for any length of time
which is sufficient to cause the production of the deposition product.
Preferably the second contacting step is performed for a sufficient
time in order to maximize the yield of the deposition product.
Preferably the second contacting step is performed for between about 1
minute and about 60 minutes.

The first contacting step may be performed at any temperature. The
second contacting step may be performed at any temperature.
Preferably, however, the second contacting step is performed at a
temperature of between about 2 degrees Celsius and about 60 degrees
Celsius.

The method according to the first aspect may be further comprised of
the step of washing the composite material following the second
contacting step.

The method according to the first aspect may be further comprised of
the step of adding an amount of an oxidizing agent to the second basic
environment during the second contacting step. The oxidizing agent may
be comprised of any oxidizing agent which is compatible with the
metal, but the oxidizing agent is preferably selected from the group
of oxidizing agents consisting of persulfates, permanganates,
peroxides and mixtures thereof. More preferably the oxidizing agent is
comprised of a persulfate. The persulfate may be comprised of any
persulfate but preferably the persulfate is selected from the group of
persulfates consisting of potassium persulfate, sodium persulfate,
ammonium persulfate and mixtures thereof. More preferably the
persulfate is comprised of ammonium persulfate, potassium persulfate
or mixtures thereof, and most preferably the persulfate is comprised
of potassium persulfate.

The amount of the oxidizing agent is preferably selected to be
compatible with the amount of the ions of the metal so that the
deposition product can be produced as efficiently as possible. In
other words, the amount of the oxidizing agent is preferably selected
to be a stoichiometrically appropriate amount relative to the amount
of the ions of the metal. Preferably the amount of persulfate
oxidizing agent is selected such that a concentration of the
persulfate in the second basic solution is between about 1 gram per
liter and about 25 grams per liter.

The method according to the first aspect may be further comprised of
the step, prior to the first contacting step, of etching the substrate
by immersing the substrate in an etching solution in order to prepare
the substrate for the deposition product. The etching step may involve
either or both of a physical process or a chemical process. The
etching step preferably prepares the substrate for the deposition
product by increasing the roughness of the substrate surface and/or
creating attraction sites for adsorption and/or deposition of the
deposition product.

Any etching solution may be utilized which is suitable for a
particular substrate. For example, where the substrate is comprised of
an organic material or polymer such as polyethylene, the etching
solution is preferably comprised of a mixture of an alcohol and an
aqueous solution of a hydroxide compound. The hydroxide compound may
be comprised of any hydroxide compound but is preferably selected from
the group of hydroxide compounds consisting of sodium hydroxide,
potassium hydroxide and mixtures thereof. More preferably the
hydroxide compound is comprised of sodium hydroxide. The etching step
may be performed for any length of time sufficient to prepare the
substrate, but preferably the etching step is performed for less than
about 20 minutes and preferably is performed for at least 5 minutes.

The method according to the first aspect may be further comprised of
the step of adding a residual silver compound to the second basic
environment during the second contacting step. The residual silver
compound may be comprised of any suitable source of silver ions, but
preferably the residual silver compound is comprised of silver
nitrate. Preferably the amount of residual silver compound is selected
such that a concentration of the residual silver compound in the
second basic solution is between about 1 gram per liter and about 5
grams per liter.

The method according to the first aspect may be further comprised of
the step of agitating the second basic environment during at least a
portion of the second contacting step in order to enhance the
production of the deposition product and the composite material.

In a second aspect, the invention is a method for producing a
deposition product, wherein the deposition product is comprised of at
least one oxidized species of a metal, the method comprising the
following steps: (a) providing a deposition solution comprising an
amount of ions of the metal and an amount of an oxidizing agent; and
(b) producing the deposition product by facilitating a chemical
reaction in the deposition solution between the ions of the metal and
the oxidizing agent.

The metal may be comprised of any metal or combinations of metals but
preferably the metal is comprised of silver so that the ions of the
metal are comprised of silver ions. The deposition solution may be
comprised of silver ions from any source or in any form but preferably
the deposition solution is comprised of an aqueous solution of a
silver salt. More preferably the silver salt is comprised of silver
nitrate.

The ions of the metal may be present in any concentration. Preferably,
where the ions of the metal are comprised of silver ions, the amount
of the silver ions is selected so that a concentration of the silver
salt in the deposition solution is between about 1 gram per liter and
about 20 grams per liter.

The oxidizing agent may be comprised of any oxidizing agent which is
compatible with the metal, but the oxidizing agent is preferably
selected from the group of oxidizing agents consisting of persulfates,
permanganates, peroxides and mixtures thereof. More preferably the
oxidizing agent is comprised of a persulfate. The persulfate may be
comprised of any persulfate but preferably the persulfate is selected
from the group of persulfates consisting of potassium persulfate,
sodium persulfate, ammonium persulfate and mixtures thereof. More
preferably the persulfate is comprised of ammonium persulfate,
potassium persulfate or mixtures thereof, and most preferably the
persulfate is comprised of potassium persulfate.

The amount of the oxidizing agent is preferably selected to be
compatible with the amount of the ions of the metal so that the
deposition product can be produced as efficiently as possible. In
other words, the amount of the oxidizing agent is preferably selected
to be a stoichiometrically appropriate amount relative to the amount
of the ions of the metal. For example, where the metal is comprised of
silver nitrate the amount of silver nitrate is preferably selected
such that a concentration of the silver nitrate in the deposition
solution is between about 1 gram per liter and about 20 grams per
liter, in which case the amount of the oxidizing agent is preferably
selected so that a concentration of the oxidizing agent in the
deposition solution is between about 1 gram per liter and about 50
grams per liter.

The method according to the second aspect may be used to produce a
deposition product which comprises a substance having the general
formula Ag.sub.7O.sub.8X, where X is an anion. The deposition product
may be further comprised of Ag.sub.2SO.sub.4. The deposition product
may also be comprised of other oxidized silver compounds such as one
or more silver oxides selected from the group of silver oxides
consisting of monovalent silver oxide, bivalent silver oxide,
trivalent silver oxide and mixtures thereof.

The anion X may be comprised of a single anion or may be comprised of
a plurality of different anions. The anion may therefore be comprised
of any anion or combination of ions. The anion may, for example, be
selected from the group of anions consisting of HCO.sub.3.sup.-,
CO.sub.3.sup.2-, NO.sub.3.sup.-, ClO.sub.4.sup.-, SO.sub.4.sup.2-,
F.sup.-, and mixtures thereof. The source of the anion may be a metal
compound which provides the ions of the metal. For example, where the
deposition solution is comprised of a silver salt such as silver
nitrate, the anion may be comprised of the nitrate ion (NO.sub.
3.sup.-). An alternative or secondary source of the anion X may
optionally be provided in order to provide sufficient quantities of
the anion for production of the deposition product.

As a result, in the method according to the second aspect, the method
may be further comprised of the step of adding a source of anions to
the deposition solution. The source of anions may be comprised of one
or more acids. The acid may be comprised of any organic or inorganic
acid. For example, the acid may be selected from the group of acids
consisting of carbonic acid, nitric acid, perchloric acid, sulfuric
acid, acetic acid, fluoroboric acid, phosphoric acid, phosphorous
acid, citric acid, acetylsalicylic acid and mixtures thereof. The
amount of the source of anions which is added to the deposition
solution preferably is an amount which is selected to be compatible
with the amount of the ions of the metal. In other words, the amount
of the source of anions is preferably selected to be a
stoichiometrically appropriate amount relative to the amount of the
ions of the metal.

The deposition product producing step is preferably performed at a
relatively low temperature, since the deposition product may
experience increasing solubility with increasing temperature. The
deposition product producing step is preferably performed at a
temperature of between about 2 degrees Celsius and about 60 degrees
Celsius, more preferably at a temperature of between about 2 degrees
Celsius and about 40 degrees Celsius, and even more preferably at a
temperature of between about 10 degrees Celsius and about 25 degrees
Celsius.

Preferably the deposition solution is agitated during at least a
portion of the deposition product producing step in order to enhance
the production of the deposition product.

The method according to the second aspect may be used to produce the
deposition product as a product, or may be used to produce a composite
material comprising a substrate and the deposition product. Where the
method is used to produce a composite material, the method may be
further comprised of the following steps: (a) providing a substrate;
and (b) contacting the substrate with the deposition solution during
the deposition product producing step, thereby producing a composite
material comprising the substrate and the deposition product.

The substrate contacting step may be performed for any length of time
which is sufficient to produce the composite material having a desired
composition. The substrate contacting step is preferably performed for
at least about 1 minute, more preferably for between about 1 minute
and about 60 minutes, even more preferably for between about 1 minute
and about 20 minutes, and even more preferably for between about 2
minutes and about 10 minutes.

The method in the second aspect may be further comprised of the step,
following the substrate contacting step, of washing the composite
material.

The method according to the second aspect may be further comprised of
the step, prior to the substrate contacting step, of etching the
substrate by immersing the substrate in an etching solution in order
to prepare the substrate for the deposition product. The etching step
may involve either or both of a physical process or a chemical
process. The etching step preferably prepares the substrate for the
deposition product by increasing the roughness of the substrate
surface and/or creating attraction sites for adsorption and/or
deposition of the deposition product.

Any etching solution may be utilized which is suitable for a
particular substrate. For example, where the substrate is comprised of
an organic material or polymer such as polyethylene, the etching
solution is preferably comprised of a mixture of an alcohol and an
aqueous solution of a hydroxide compound. The hydroxide compound may
be comprised of any hydroxide compound but is preferably selected from
the group of hydroxide compounds consisting of sodium hydroxide,
potassium hydroxide and mixtures thereof. More preferably the
hydroxide compound is comprised of sodium hydroxide. The etching step
may be performed for any length of time sufficient to prepare the
substrate, but preferably the etching step is performed for less than
about 20 minutes and preferably is performed for at least 5 minutes.
Where the etching step is performed, the method according to the
second aspect preferably further comprises the step, following the
etching step, of washing the substrate to remove residual alkali from
the substrate.

The method according to the second aspect may be further comprised of
the step, following the substrate contacting step, of immersing the
composite material in boiling water. The immersing step may be useful
for converting the deposition product into other oxidized silver
species (such as silver oxides), thus potentially providing an
opportunity further to "engineer" the composite material to provide
desired properties of the deposition product. The immersing step may
be performed for any length of time, but preferably the immersing step
is performed for at least about 1 minute.

The composite material may be produced for many different applications
including for electronics, materials engineering and medical purposes.
The method according to the second aspect is particularly suited for
the production of medical devices in circumstances where the metal is
silver and the deposition product is comprised of an oxidized silver
species having the general formula Ag.sub.7O.sub.8X and optionally
Ag.sub.2SO.sub.4 and/or optionally one or more silver oxide compounds,
due to the antimicrobial properties exhibited by the deposition
product and to the capability to control the extent of the deposition
of the deposition product on the substrate.

The term "medical device" as used herein means any article which has a
medical application where antimicrobial properties may be desirable,
and includes all natural and synthetic materials and both fibrous and
non-fibrous materials. For example, the materials may be comprised of
a metal, plastic, paper, glass, ceramic, textile, rubber, polymer,
composite material or any other material or combination of materials.
Non-limiting examples of medical devices which are encompassed by the
invention include wound dressings, splints, sutures, catheters,
implants, tracheal tubes, orthopedic devices, drains, shunts,
connectors, prosthetic devices, needles, medical instruments,
laboratory, clinic and hospital equipment, furniture and furnishings,
dental devices, as well as health care products such as personal
hygiene products, sterile packaging, clothing, footwear etc.

Accordingly, the composite material may comprise a medical device or a
component of a medical device and the term "medical device" as used
herein extends to both medical devices and components of medical
devices.

In a preferred embodiment, the substrate is comprised of a wound
dressing. The wound dressing may be comprised of any material or
combination of materials, including but not limited to metals,
ceramics, glass, polymers, plastics, composite materials, natural
materials, synthetic materials, synthetic textiles such as HDPE,
rayon, nylon, polyacetates, polyacrylics and glass and natural
textiles such as cellulose, wool, jute and cotton, whether in fibrous
or non-fibrous form.

In a preferred embodiment of wound dressing, the wound dressing may be
comprised of a polymer material such as high density polyethylene and
may be further comprised of an adhesive material comprising a skin
adhesive layer. The skin adhesive layer may be comprised of a cross-
linked silicon gel material. The wound dressing and/or the cross-
linked silicon gel material may for example be comprised of a product
sold under the Mepitel.TM. trade-mark or the Safetac.TM. trade-mark,
both of which trade-marks are owned by Molnlycke Health Care AB of
Sweden.

In one application, the deposition product may be selectively
deposited on the skin adhesive layer and the production of the
deposition product is preferably controlled so that the deposition
product does not materially interfere with the adhesive properties of
the skin adhesive layer, yet still provides an acceptable
antimicrobial effect without significant undesirable toxic effects.
This result may be achieved by depositing the deposition product on
the skin adhesive layer such that the deposition product provides a
desired antimicrobial effect but does not completely cover the surface
of the skin adhesive layer. In this application, preferably the amount
of the deposition product which is deposited on the substrate is such
that the amount of total silver on the substrate is selected to be
between about 0.1 mg/cm.sup.2 and about 1.0 mg/cm.sup.2, or more
preferably between about 0.2 mg/cm.sup.2 and about 0.6 mg/cm.sup.2, in
order to achieve the desired result.

In other applications in which the deposition product is not deposited
on an adhesive such as the skin adhesive layer, the amount of the
deposition product is preferably controlled to balance the desired
antimicrobial effect, undesirable toxic effects, and economic
considerations.

In a third aspect, the invention is a medical device comprising a
composite material, wherein the composite material is comprised of a
substrate and a deposition product and wherein the deposition product
is comprised of an antimicrobially active oxidized silver species
comprising a silver salt and a silver oxide.

The medical device according to the third aspect may be produced using
any of the methods of the invention. Preferably the medical device is
produced using a method according to the second aspect of the
invention.

In certain preferred embodiments the invention provides methods for
depositing a deposition product comprising at least one oxidized
silver species onto a substrate, thus producing a composite material.
Since the oxidized silver species of the invention exhibit an
antimicrobial activity, composite materials comprising the oxidized
silver species can be used in various medical devices for prevention
or inhibition of infections. These medical devices may include but are
not limited to wound dressings, adhesives, sutures, catheters and
other articles where antimicrobial properties are desirable.

The preferred embodiments of the invention may be used to produce
deposition products and composite materials from aqueous solutions
under a wide range of pH conditions, involving reactions in either
acidic or alkaline solutions. The methods can be performed at, but are
not limited to, temperatures between about 2 degrees Celsius and about
60 degrees Celsius with about 10 degrees Celsius to about 40 degrees
Celsius being the most preferable.

The method steps for certain preferred embodiments of the invention
are as follows:

I. Under acidic conditions:

(a) immersing an article to be used as a medical device in an aqueous/
alcohol solution of NaOH for a sufficient time to provide a reasonable
etching and cleaning of the surface, followed by washing of the
article with distilled water until a pH of 7 is attained, in order to
remove residual alkali; (b) immersing the article in an aqueous silver
salt solution. The aqueous silver salt solution may be prepared from
any silver salt which is soluble in water with the most preferred
silver salt being silver nitrate; (c) adding a stoichiometrically
suitable quantity of an oxidizing agent to the mixed silver salt
solution containing the article. The oxidizing agent can be any
oxidizing substance such as persulfates, permanganates, hydrogen
peroxide and the like, with potassium persulfate (K.sub.2S.sub.2O.sub.
8) being the most preferred oxidizing agent; (d) adding a
stoichiometrically suitable quantity of an acid to the mixed silver
salt solution containing the immersed article in order to provide a
source of anions. The acids that can be used include any inorganic or
organic acids including, but not limited to carbonic acid, nitric
acid, perchloric acid, sulfuric acid, acetic acid, fluoroboric acid,
phosphoric acid, phosphorous acid, citric acid, acetylsalicylic acid
and mixtures thereof, but most preferably nitric acid, perchloric
acid, phosphoric acid, acetic acid or sulfuric acid; (e) agitating the
article in the mixed silver salt solution comprising the soluble
silver salt (preferably AgNO.sub.3), the acid (preferably nitric acid,
perchloric acid, phosphoric acid, acetic acid or sulfuric acid), and
the oxidizing agent (preferably potassium persulfate) at temperatures
between 2 degrees Celsius and 30 degrees Celsius with temperatures
between 10 degrees Celsius and 25 degrees Celsius being the most
preferred for between about 2 and 40 minutes until the article is
coated with a grayish, gray or black color; (f) removing the article
from the slurry and washing the article with distilled water until a
pH of 7 is achieved; and (g) drying the article at room temperature.

Alternatively after step (e) the article may be immersed in boiling
water (about 90 degrees Celsius to about 100 degrees Celsius) for at
least 1 minute.

II. Under Alkaline Conditions:

(a) immersing an article to be used as a medical device in an aqueous/
alcohol solution of NaOH for a sufficient time to provide a reasonable
etching and cleaning of the surface; (b) removing the article into a
solution containing a silver diamino complex in a concentration
sufficient to adsorb the silver ions at the surface of the article and
for a duration of about 2 minutes to about 5 minutes. The silver
diamino complex may be prepared by dissolving any silver salt or
silver oxide in ammonium hydroxide, and may be achieved by adding a
stoichiometrically suitable quantity of ammonium hydroxide to an
aqueous solution or suspension of the silver salt or silver oxide
until a clear colorless solution containing [Ag(NH.sub.3).sub.2].sup.+
is obtained. The pH of this solution is usually in the range from
about 8 to about 12; (c) removing the article without washing or
rinsing into another solution containing a strong alkali, most
preferably NaOH or KOH, and agitating the article in this solution
until a clear colorless solution is obtained and the article is
clearly dyed with a tan, gray, brown or black color, depending on the
desired amount of oxidized silver species. The time of contact of the
article with the alkaline solution may vary, depending on temperature
and silver ion concentration, but the most preferable duration is
about 1 minute to about 15 minutes at room temperature or about 1
minute to about 10 minutes at a temperature of between about 40
degrees Celsius and about 60 degrees Celsius; (d) removing the dyed
article from the solution and washing with distilled water until a pH
of 7 is achieved; and (e) drying the article at room temperature.

Alternatively, in step (c), the method may involve, depending on the
amount of silver required at the surface of the article, further
additions to the strong alkali solution of the silver diamino complex
solution and/or additions to the strong alkali solution of an
oxidizing agent such as a persulfate, permanganate, peroxide or a
mixture thereof, with potassium persulfate being the most preferred
oxidizing agent.

BRIEF DESCRIPTION OF DRAWINGS

Embodiments of the invention will now be described with reference to
the accompanying drawings, in which:

FIG. 1 is an XRD pattern generated from a deposition product obtained
from the reaction of AgNO.sub.3 and (NH.sub.4).sub.2S.sub.2O.sub.8
according to Examples 15-16.

FIG. 2 is an SEM micrograph (magnification=2000.times.) generated from
a deposition product obtained from the reaction of AgNO.sub.3 and
(NH.sub.4).sub.2S.sub.2O.sub.8 according to Examples 15-16.

FIG. 3 is an XRD pattern generated from a deposition product obtained
from the reaction of AgNO.sub.3 and K.sub.2S.sub.2O.sub.8 according to
Examples 15-16.

FIG. 4 is an SEM micrograph (magnification=2000.times.) generated from
a deposition product obtained from the reaction of AgNO.sub.3 and
K.sub.2S.sub.2O.sub.8 according to Examples 15-16.

FIG. 5(a) is an SEM micrograph (magnification=150.times.) generated
from a sample of uncoated HDPE mesh.

FIG. 5(b) is an SEM micrograph (magnification=1000.times.) generated
from a sample of HDPE mesh upon which a deposition product has been
deposited according to Examples 15-16.

FIG. 6(a) is a photograph depicting a controlled zone of inhibition
(CZOI) against Staphylococcus Aureus for a sample of HDPE mesh coated
with a deposition product according to Examples 15-16.

FIG. 6(b) is a photograph depicting a controlled zone of inhibition
(CZOI) against Pseudomonas Aeruginosa for a sample of HDPE mesh coated
with a deposition product according to Examples 15-16.

FIG. 6(c) is a photograph depicting a controlled zone of inhibition
(CZOI) against Candida Albicans for a sample of HDPE mesh coated with
a deposition product according to Examples 15-16.

FIG. 7 is an SEM micrograph (magnification=30.times.) generated from a
substrate consisting of an uncoated sample of a perforated plastic
carrier material with a skin adhesive layer comprised of a hydrophobic
cross-linked silicon gel (trade-mark Mepitel.TM.).

FIG. 8 is an SEM micrograph (magnification=40.times.) generated from a
composite material consisting of a coated sample of a perforated
plastic carrier material with a skin adhesive layer comprised of a
hydrophobic cross-linked silicon gel (trade-mark Mepitel.TM.), in
which a relatively small amount of deposition product has been
deposited on the substrate in accordance with the second and third
aspects of the invention.

FIG. 9 is an SEM micrograph (magnification=2000.times.) generated from
the composite material of FIG. 8, depicting the density and coverage
of the deposition product on the substrate.

FIG. 10 is an SEM micrograph (magnification=40.times.) generated from
a composite material consisting of a coated sample of a perforated
plastic carrier material with a skin adhesive layer comprised of a
hydrophobic cross-linked silicon gel (trade-mark Mepitel.TM.), in
which a relatively larger amount of deposition product (relative to
FIG. 8 and FIG. 9) has been deposited on the substrate in accordance
with the second and third aspects of the invention.

FIG. 11 is an SEM micrograph (magnification=2000.times.) generated
from the composite material of FIG. 10, depicting the density and
coverage of the deposition product on the substrate.

FIG. 12 is an XRD pattern generated from a substrate consisting of an
uncoated sample of a perforated plastic carrier material with a skin
adhesive layer comprised of a hydrophobic cross-linked silicon gel
(trade-mark Mepitel.TM.).

FIG. 13 is an XRD pattern generated from a composite material
consisting of a coated sample of a perforated plastic carrier material
with a skin adhesive layer comprised of a hydrophobic cross-linked
silicon gel (trade-mark Mepitel.TM.), in which a deposition product
has been deposited on the substrate in accordance with the second and
third aspects of the invention.

FIG. 14 is a superimposition of the XRD patterns depicted in FIG. 12
and FIG. 13.

DETAILED DESCRIPTION

In preferred embodiments of the invention, antimicrobial properties of
medical devices are achieved by the adsorption and deposition of a
deposition product comprising an antimicrobially active silver species
within or at the surface of the medical device. These active silver
species may include but are not limited at all oxidized silver species
such as silver salts, silver oxide (Ag.sub.2O), higher silver oxides
i.e. Ag(II) and Ag(III) (AgO, Ag.sub.2O.sub.3, Ag.sub.3O.sub.4 or
like), silver oxy-salts with a general formula Ag.sub.7O.sub.8X where
X can include one of acid anions such as sulfates, chlorides,
phosphates, carbonates, citrates, tartrates, oxalates and like. The
deposition product may also contain some elemental silver deposited
during the process.

The term "total silver" as used in herein is the total amount of
silver as determined by a chemical analysis, which may include
elemental (metallic) silver as well as silver originating from
oxidized silver species.

The term "oxidized silver species" as used herein may involve but is
not limited at all compounds of silver where said silver is in +I, +II
or +III valent states or any combinations thereof. These oxidized
silver species include, for example silver (I) oxide, silver (II)
oxide, silver (III) oxide or mixtures thereof, all silver salts having
a solubility product higher than 10.sup.-20 (such as for example
Ag.sub.2SO.sub.4, AgCl, Ag.sub.2S.sub.2O.sub.8, Ag.sub.2SO.sub.3,
Ag.sub.2S.sub.2O.sub.3, Ag.sub.3PO.sub.4, and the like), and silver
oxy-salts such as Ag.sub.7O.sub.8X were X can include but is not
limited at NO.sub.3.sup.-, ClO.sub.4.sup.-, SO.sub.4.sup.2-, F.sup.-
etc.

The term "medical device materials" as used herein may include
materials such as metals, ceramics, glass, polymers, plastics,
composite materials, a variety of natural materials, fabrics, textile
made of either synthetic (HDPE, rayon, nylon, polyacetates,
polyacrylics, glass etc.) or natural (cellulose, wool, jute, cotton,
etc.) fibers.

The term "bacteriostatic activity", as used herein relates to the
inhibition of bacterial growth, but not to actually killing the
bacteria. Successful treatment therefore requires the host's immune
system to clear the pathogen. Treatment is compromised when the
antimicrobial materials are stopped before the pathogen has been
completely cleared.

The term "bactericidal activity" as used herein relates to killing
bacteria with or without lysis of the target cell. These types of
antimicrobial materials are particularly advantageous in
immunosuppressed individuals. A disadvantage to bactericidal activity
is cell lysis, which can release lipolysaccharides which are toxic to
the host. However, if the concentration of the said antimicrobial
material is relatively low so that toxic effects cannot occur, a
combination of both bacteriostatic and bactericidal activities may be
ideal for antimicrobial materials.

In the preferred embodiments, the deposition of the deposition product
comprising the oxidized silver species is accomplished by first
providing an aqueous solution of monovalent silver salt or a silver
complex such as silver nitrate, perchlorate or silver diamino complex,
with silver nitrate being the most preferable if the reaction is
carried out under acidic conditions or at close to neutral conditions
(i.e. at pH below 7), and with silver diamino complex, (i.e.,
[Ag(NH.sub.3).sub.2].sup.+) being the most preferable if the reaction
is carried out under alkaline conditions (i.e. at pH above 7).

Prior to the production of the composite material comprising the
article as a substrate and the deposition product, the article is
preferably immersed in an alkaline solution containing 50 vol. %
ethanol and 50 vol. % of an aqueous solution containing 30 g/L NaOH.
Other cleaning and etching solutions can be used depending upon the
material from which the medical device is made, upon the toxicity of
the said cleaning or etching solutions, and upon the possibility that
some toxic substances may adsorb at the surface of the article. Of
course any use of toxic or carcinogenic substances during the etching
step should be avoided. If production of the deposition product is
carried out under acidic conditions, the article is preferably washed
with distilled water after the etching step until a pH of 7 is
achieved in order to remove residual alkali remaining after the
etching step.

When the reaction is carried out in the pH range below 7 (i.e., under
acidic conditions), the clean pretreated article to be used as a
medical device containing oxidized silver species at the surface of
the same is simply immersed into an agitated 1% AgNO.sub.3 aqueous
solution as a deposition solution. After exposure of the said article
to the deposition solution for a duration preferably of about 2 to
about 10 minutes, a solution of an oxidizing agent is added.
Alternatively, the oxidizing agent may be added to the deposition
solution before the article is immersed into the deposition solution,
but this may result in some production of the deposition product
before the article is present in the deposition solution.

Although a wide range of oxidizing agents such as permanganates,
persulfates, hydrogen peroxide, hypochlorites etc., may be used under
specific conditions and with the proper concentrations, the preferred
oxidizing agent is a persulfate, more preferably either ammonium
persulfate or potassium persulfate., and most preferably potassium
persulfate The persulfate facilitates the precipitation and deposition
of the deposition product on or within the article.

The concentration of persulfate in the deposition solution may be in a
range from about 1 gram per liter to about 250 gram per liter with the
concentration of about 50 gram per liter being the most preferable.
After agitation for about 2 minutes to about 5 minutes, the solution
of 1% AgNO.sub.3 and persulfate may be acidified with an organic or
inorganic acid such as HNO.sub.3, HClO.sub.4, H.sub.2SO.sub.4 or
CH.sub.3COOH such that the concentration of the free acid preferably
is about 9% HNO.sub.3, 9% HClO.sub.4 acid, 5% H.sub.2SO.sub.4, or 5%
CH.sub.3COOH. Although other acids may be used the most preferable
acids are H.sub.2SO.sub.4, HClO.sub.4 or HNO.sub.3.

The agitation of the deposition solution is not strictly required, but
in order to achieve a more uniform distribution of the deposition
product and an efficient reaction yield, the agitation of the solution
is recommended. Agitation can be realized by many different ways such
as for example mechanical stirring, magnetic stirring or ultrasonic
agitation.

Following addition of the persulfate (preferably potassium persulfate)
to the deposition solution of 1% AgNO.sub.3 within the time of about 1
minute to about 10 minutes, and depending on the concentration of the
persulfate as well as on the conditions of agitation, the formation
first of a yellow brown color of the solution and then a black grayish
precipitate will occur. This brown color of the solution is attributed
to the oxidation of Ag(I) to Ag(II).

The black grayish deposit at the article or in the bulk solution is a
consequence of the formation of silver oxy-salts such as Ag.sub.7O.sub.
8X, were X is an anion, depending on the acid used in the method e.g.
HNO.sub.3 (NO.sub.3.sup.-), H.sub.2SO.sub.4 (SO.sub.4.sup.2-), etc.
The decomposition of the silver oxy-salts may be presented as:
Ag(Ag.sub.3O.sub.4).sub.2X=AgX+AgO (1)

Persulfates are powerful oxidizing agents. They can therefore be
reduced in aqueous solutions according to the following reactions:
S.sub.2O.sub.8.sup.2-+2e.sup.-=2SO.sub.4.sup.2-, with E.degree.=1.96 V
(2) S.sub.2O.sub.8.sup.2-+2H.sup.++2e.sup.-=2HSO.sub.4.sup.-, with
E.degree.=1.96 V (3) and S.sub.2O.sub.8.sup.2-+2H.sub.2O=2H.sup.+
+2SO.sub.4.sup.2-+H.sub.2O.sub.2, with .DELTA.G.degree.=-36 kJ/mol (4)

A consequence of the reduction of persulfate is the oxidation of Ag(I)
to Ag(II) and Ag(III), probably according to the following reactions:
Ag.sup.+=Ag.sup.2++e.sup.-, with E.degree.=1.98 V (5) Ag.sup.++H.sub.
2O=AgO.sup.++2H.sup.++e.sup.-, with E.degree.=1.998 V (6) Ag.sup.2+
+H.sub.2O=AgO.sup.++2H.sup.++e.sup.-, with E.degree.=2.06 V (7) Ag.sup.
++H.sub.2O=AgO+2H.sup.++e.sup.-, with E.degree.=1.772 V (8)

In this way the composite material comprising the article to be used
as a medical device and the deposition product may include a
combination of oxidized silver species i.e. Ag(I)-- and Ag(II)--
oxides as well as silver salts such as nitrates, persulfates,
sulfates, phosphates, perchlorates and like, silver salts of a general
formula Ag.sub.7O.sub.8X and perhaps traces of pure elemental silver.
After production of the composite material, the article is removed
from the deposition solution and then preferably washed with distilled
water until a pH of 7 is achieved. When the washing is completed, the
medical device comprising the composite material may be dried at room
temperature and packaged.

When the reaction is carried out in the pH range above 7 (i.e., under
alkaline conditions) the article to be used as a medical device is
first immersed in an etching solution comprising an alkaline solution
containing alcohol. The most preferable solution according to this
invention is either NaOH or KOH with concentrations 15 to 40 g/L. The
alcohol used in this solution may be ethyl alcohol, methyl alcohol or
mixtures therein in a concentration above 50 vol. %. The immersion of
the article into the etching solution is carried out in order to etch
and clean the surface of the article to provide a reasonable adhesion
of the deposition product comprising an oxidized silver species which
is deposited on or within the article thereafter. The immersion time
of the article is preferably in the range of between about 5 minutes
and about 20 minutes, with about 10 minutes being the most preferable.

After the exposure to the alkali/alcohol solution for about 10
minutes, the article is then removed without washing or rinsing into a
first basic environment comprising a first basic solution containing
silver diamino complex i.e. [Ag(NH.sub.3).sub.2].sup.+ in a
concentration sufficient to adsorb silver ions at the surface of the
article and for a duration of about 2 minutes to about 5 minutes. The
silver diamino complex is preferably prepared from a silver salt or
silver oxide dissolved or suspended in water by a dissolution with
NH.sub.4OH (28 vol. %).

Consequently, the first basic solution is prepared in a way such that
a solution of any silver salt (such as for example AgNO.sub.3 or
AgClO.sub.4) or any silver oxide (such as Ag.sub.2O or Ag.sub.2O.sub.2
or AgO) or any silver salt suspended in water (such as AgCl, Ag.sub.
2CO.sub.3, Ag.sub.2SO.sub.4 or the like), the ammonium hydroxide is
added in a stoichiometrically suitable concentration so that a clear
colorless solution is obtained. The concentration of silver ion in
this silver diamino complex solution, as calculated for Ag.sup.+ ion
can vary from 1 to 20 g/L with about 10 g/L being the most preferable.
The pH of the first basic solution is usually between about 8 and
about 12 with the most preferred pH being in the range of between
about 10 and about 11.

After exposure of the article to the first basic solution for about 2
minutes to about 5 minutes, the article is removed without washing or
rinsing into a second basic environment comprising a second basic
solution containing a strong alkali, most preferably NaOH or KOH. The
article is kept in this solution under agitation until a clear
colorless solution is obtained and the article is dyed with a tan,
gray, brown or black color, depending on the desired amount of
oxidized species to be deposited at or within the surface of the
article. The time of contact of the article with the second basic
solution may vary depending on temperature and the silver ion
concentration, but most preferable time is about 1 minute to about 15
minutes at room temperature or about 1 minute to about 10 minutes at a
temperature of between about 40 degrees Celsius and about 60 degrees
Celsius.

Alternatively, the method may involve an addition of an oxidizing
agent to the second basic solution, preferably a persulfate, more
preferably either ammonium persulfate or potassium persulfate, and
most preferably potassium persulfate. The oxidizing agent may be added
directly to the second basic solution containing the article. In
addition, depending on the amount of silver desired to be deposited as
the deposition product, addition of a residual silver compound such as
the silver diamino complex [Ag(NH.sub.3).sub.2].sup.+ may also be
beneficial.

Upon immersion of the article, previously exposed to the first basic
solution, into the second basic solution, the following reaction at
the surface of the article may occur: 2Ag(NH.sub.3).sub.2NO.sub.
3+2NaOH=Ag.sub.2O+4NH.sub.3+H.sub.2O+2NaNO.sub.- 3 (9)

In this way, at the surface of the article, Ag.sub.2O will deposit as
the result of the reaction (9). The addition of an oxidizing agent
such as ammonium persulfate (i.e., (NH.sub.4).sub.3S.sub.2O.sub.8) to
the second basic solution may result in the oxidation of silver ions
and the reduction of S.sub.2O.sub.8.sup.2- ions pursuant to the
following reactions: Ag.sup.+=Ag.sup.2++e.sup.-, with E.degree.=1.96 V
(10) and S.sub.2O.sub.8.sup.2-+2e.sup.-=2SO.sub.4.sup.2-, with
E.degree.=1.96 V (11)

The reactions of Ag(NH.sub.3).sub.2.sup.+ ion with ammonium persulfate
can be represented as follows: Ag(NH.sub.3).sub.2NO.sub.3+(NH.sub.
4).sub.2S.sub.2O.sub.8=Ag.sub.2S.sub.2- O.sub.8+2NH.sub.4NO.sub.
3+4NH.sub.3 (12) Ag.sub.2S.sub.2O.sub.8+H.sub.2O=2AgO+2H.sub.2SO.sub.4
(13) Ag(NH.sub.3).sub.2NO.sub.3+(NH.sub.4).sub.2S.sub.2O.sub.8+2H.sub.
2O=2NH.s- ub.4NO.sub.3+2AgO+2H.sub.2SO.sub.4+4NH.sub.3 (14) or
Ag(NH.sub.3).sub.2NO.sub.3+(NH.sub.4).sub.2S.sub.2O.sub.8+2H.sub.
2O=2NH.s- ub.4NO.sub.3+2AgO+2(NH.sub.4).sub.2SO.sub.4 (15)

In this way, the deposition product may contain Ag.sub.2O, AgO or
other higher oxides of silver Ag(II), Ag(III) and mixtures therein.
Also, if alcohol is present in the reacting solution, due to
transferring from the etching solution some elemental silver may occur
in the deposition product. This is because in the presence of
persulfates, alcohols can be oxidized to aldehydes according to the
reactions: CH.sub.3OH=H.sub.2CO+2H.sup.++2e.sup.- (16) C.sub.2H.sub.
5OH=CH.sub.3CHO+2H.sup.++2e.sup.- (17)

Under the alkaline conditions, the aldehydes can reduce the silver
ions to the elemental silver according to the reaction: 2Ag(NH.sub.
3).sub.2OH+HCHO=2Ag+4NH.sub.3+HCOOH+H.sub.2O (18)

After production of the composite material comprising the article and
the deposition product comprising the oxidized silver species is
completed, the article is removed, carefully washed with water until a
pH of 7 is achieved. The article may then be dried at room temperature
and packaged.

Following are examples which illustrate the present invention.

EXAMPLES

Example 1

Nine (9) pieces of high density polyethylene mesh (HDPE), with
dimensions 10.times.8 cm each, were immersed in 100 ml of an etching
solution containing 50 mL alcohol (95% C.sub.2H.sub.5OH and 5% CH.sub.
3OH) and 50 mL of 28 g/L NaOH solution for 5 minutes. After 5 minutes
of etching the HDPE mesh was transferred without washing or rinsing
into 40 mL of an Ag.sup.+ solution, containing 15.3 g/L AgNO.sub.3 and
a stoichiometrically suitable volume of NH.sub.4OH (28 vol. %). The
HDPE mesh was kept in this solution for 2 minutes. After 2 minutes of
exposure to the ammoniacal Ag(NH.sub.3).sub.2+solution, the HDPE mesh
was transferred without washing or rinsing into 150 mL of a 28 g/L
NaOH solution stirred with a magnetic stirrer. As soon as the HDPE
mesh was immersed into NaOH solution, the formation of a precipitate
yellowish-brown in color occurred. Under agitation a residual silver
compound (about 38 mL of the Ag(I) solution) was added and after that
5 mL of a 250 g/L (NH.sub.4).sub.2S.sub.2O.sub.8 solution was added.
Agitation was continued for 10 minutes. During this time the solution/
precipitate became black. The HDPE mesh was uniformly coated and was
black and shiny in appearance. The coated HDPE mesh was then removed
from the solution and carefully washed with distilled water until pH
7.00, and dried at room temperature. After drying, the mesh was a
black and shiny in appearance.

Chemical analysis determined that the HDPE mesh coated with oxidized
silver species contained about 0.08 mg total silver per cm.sup.2 of
mesh. The coated mesh was further analyzed by XRD analysis. As found
by the XRD analysis the mesh included Ag.sub.2O, Ag(II) oxides, Ag.sub.
7O.sub.8NO.sub.3 and some traces of the elemental silver. Both
bacteriostatic and bactericidal activities of silver coated HDPE
substrates were tested against Pseudomonas Aeruginosa and
Staphylococcus Aureus. One hour bactericidal activity tests of coated
HDPE mesh against both Pseudomonas Aeruginosa and Staphylococcus
Aureus were positive. The bacteriostatic activity was also tested. The
controlled zone of inhibition surrounding the test sample, where no
bacteria growth occurred, was estimated at about 9 mm to about 10 mm.

Example 2

Samples of HDPE mesh with dimensions 10.times.8 cm were immersed in
100 mL of an etching solution containing 50 mL of 28 g/L NaOH and 50
mL of denatured ethanol (95% C.sub.2H.sub.5OH and 5% CH.sub.3OH) for 5
minutes. After 5 minutes of etching the HDPE mesh was transferred
without washing or rinsing into 40 mL of an ammoniacal Ag(I) solution
containing 15.3 g/L AgNO.sub.3 and a stoichiometrically suitable
quantity of NH.sub.4OH (28 vol. %). The HDPE mesh was kept in this
solution for 2 minutes. The HDPE mesh was then transferred without
washing or rinsing into 150 mL of a solution containing 28 g/L NaOH.
The NaOH solution immediately became brown. Upon addition of a
residual silver compound (about 38 mL of the Ag(I) solution) the
solution turned to a dark brown color and with a continued agitation
for about 5 minutes the solution became black. When the agitation was
stopped, the black precipitate occurred in the bulk solution as a
result of its separation from the HDPE mesh material. After washing
and rinsing with distilled water the mesh appeared to be light tan or
at the most slightly gray as a consequence of the coating with silver
compounds.

The amount of total silver deposited on the HDPE mesh as determined by
chemical analysis was estimated at about 0.04 mg/cm.sup.2.
Antimicrobial activities (bactericidal and bacteriostatic) were tested
against Pseudomonas Aeruginosa and Staphylococcus Aureus. One hour
bactericidal activity of the coated HDPE mesh was positive. The
bacteriostatic activity, as estimated according to the controlled zone
of inhibition (CZOI) for the bacterial growth was also positive. The
CZOI was estimated at about 4 mm.

Example 3

Samples of HDPE mesh were immersed in an etching solution containing
100 mL of 28 g/L NaOH solution for 5 minutes. The mesh was then
transferred without washing or rinsing into 40 mL of an ammoniacal
Ag(I) solution containing 15.3 g/L AgNO.sub.3 and a stoichiometrically
suitable volume of NH.sub.4OH (28%). After 2 minutes of immersion, the
mesh was transferred without washing or rinsing into 150 mL of a 28 g/
L NaOH solution stirred magnetically. The solution became immediately
brown due to formation of a precipitate. Addition of a residual silver
compound (about 38 mL of the Ag(I) solution) resulted in the formation
of a dark brown precipitate. The color of the solution did not change
further even after 30 minutes of mixing at room temperature. The HDPE
mesh was then washed and rinsed very carefully with distilled water.
The color of the HDPE mesh did not change significantly, but some
change in color from white to a light tan appeared.

The amount of total silver deposited on the HDPE mesh was estimated at
about 0.02 mg/cm.sup.2. The antimicrobial activities (both
bacteriostatic and bactericidal) of these samples were tested against
Pseudomonas Aeruginosa and Staphylococcus Aureus. The results showed a
positive bactericidal activity and the CZOI was estimated at about 3
mm.

Example 4

Samples of HDPE mesh were immersed in 100 mL of a solution containing
1 g AgNO.sub.3 and 1 mL of 67% HNO.sub.3 as a source of anions. After
5 minutes of immersion, 5 g of (NH.sub.4).sub.2S.sub.2O.sub.8
dissolved in 20 mL of water was added. The sample was left for 30
minutes at room temperature, during which the solution was stirred
occasionally with a glass rod. During this time the solution changed
color from colorless to a dark brown and a formation of a light gray
precipitate in the bulk solution appeared. After 30 minutes, the HDPE
mesh was removed from the solution and carefully washed with distilled
water. The washed HDPE mesh had a gray color. The coating was
uniformly distributed at the surface of this material.

The amount of total silver deposited on the HDPE mesh was estimated at
0.09 mg/cm.sup.2. The bactericidal activity for these samples was
positive. The CZOI was estimated at about 8 mm.

Example 5

HDPE mesh was coated with silver oxidized compounds using a method
similar to that described in Example 4, with a few differences as
outlined in the description that follows.

Samples of HDPE mesh were immersed in 100 mL of a solution containing
10 g/L AgNO.sub.3 and 15 mL/L HNO.sub.3 (67%) as a source of anions.
To this solution 10 mL of 500 g/L (NH.sub.4).sub.2S.sub.2O.sub.8 was
added. The solution was magnetically stirred. After 7 minutes of
stirring the solution became yellow-brown and formation of a very
small amount of precipitate occurred. The stirring was continued for
the next 30 minutes. After 30 minutes, the HDPE mesh was removed from
the slurry and carefully washed with distilled water. The washed HDPE
mesh had a gray color. The coating was uniformly distributed at the
surface of the HDPE mesh.

The amount of total silver deposited on the HDPE mesh was estimated at
0.08 mg/cm.sup.2. The bactericidal activities against Pseudomonas
Aeruginosa and Staphylococcus Aureus were positive. The CZOI was
estimated at about 7 mm.

Example 6

HDPE mesh was coated with silver oxidized compounds using a method
similar to that described in Example 4 and Example 5, with a few
differences as outlined in the description that follows.

Samples of HDPE mesh were immersed in 100 mL of a solution containing
10 g/L AgNO.sub.3 and 15 mL/L HNO.sub.3 (67%) as a source of anions.
To this solution 10 mL of 500 g/L (NH.sub.4).sub.2S.sub.2O.sub.8 was
added. The solution was agitated ultrasonically. After 2 minutes of
stirring the solution became yellow-brown and formation of a very
small amount of precipitate occurred. The stirring was continued for
the next 30 minutes. After 30 minutes, the HDPE mesh was removed from
the solution and carefully washed with distilled water. The washed
HDPE mesh had a gray color. The coating was uniformly distributed at
the surface of the HDPE mesh.

The amount of total silver deposited on the HDPE mesh was estimated at
0.08 mg/cm.sup.2. The bactericidal activities against Pseudomonas
Aeruginosa and Staphylococcus Aureus were positive. The CZOI was
estimated at about 7 mm.

Examples 7-9

In these examples the effect of different acids (i.e., sources of
anions) is clearly shown for coating of HDPE mesh with oxidized silver
species under acidic conditions. In Example 4, HNO.sub.3 was used as a
source of anions to supplement the anions contained in the AgNO.sub.3,
while in Examples 7-9 perchloric acid (HClO.sub.4), sulfuric acid
(H.sub.2SO.sub.4) and acetic acid (CH.sub.3COOH) respectively were
used as a source of anions.

Samples of HDPE mesh were immersed in 100 mL of a solution containing
1 g AgNO.sub.3. To this solution 1 mL of HClO.sub.4 (70%) (Example 7),
0.5 mL of H.sub.2SO.sub.4 (98%) (Example 8) and 15 mL of CH3COOH (5%)
(Example 9) were added. After 2 minutes of the exposure of HDPE mesh
to these solutions, 20 mL of 250 g/L (NH.sub.4).sub.2S.sub.2O.sub.8
was added. The mixing was continued for the next 30 minutes. In the
solutions containing HClO.sub.4 (Example 7) and H.sub.2SO.sub.4
(Example 8) formation of a black grayish precipitate occurred similar
to Example 4. When the precipitate settled the solutions were clear
and yellow-brown in color. The yellow-brown color suggests the
presence of Ag(II) complexes in the solution. The coated HDPE mesh was
then removed from the slurry and carefully washed and rinsed with
distilled water and thereafter dried at room temperature. After drying
the HDPE mesh coated in the presence of 1 mL of HClO.sub.4 (70%)
(Example 7), or in the presence of 0.5 mL of H.sub.2SO.sub.4 (98%)
(Example 8) appeared to be grayish in color. However, the HDPE mesh
coated in the presence of 15 mL of CH.sub.3COOH (5%) (Example 9) was
white and it did not change its color.

The coated HDPE mesh (Examples 7-9) were analyzed for the total silver
content, and the antimicrobial activity was also evaluated against
Pseudomonas Aeruginosa and Staphylococcus Aureus. The amount of total
silver deposited on the HDPE mesh was estimated at 0.08 mg/cm.sup.2
(for samples coated in the presence of HClO.sub.4), 0.07 mg/cm.sup.2
(for samples coated in the presence of H.sub.2SO.sub.4) and 0.01 mg/
cm.sup.2 (for the samples coated in the presence of CH.sub.3COOH). The
bactericidal activities against Pseudomonas Aeruginosa and
Staphylococcus Aureus were positive. The CZOI was estimated at about 6
mm (for samples coated in the presence of HClO.sub.4 or H.sub.2SO.sub.
4) and about 1 to 2 mm (for samples coated in the presence of
CH3COOH).

Example 10

Samples of HDPE mesh with dimensions 10.times.8 cm were immersed in
100 mL of an etching solution containing 50 mL of 28 g/L NaOH and 50
mL of denatured ethanol (95% C.sub.2H.sub.5OH and 5% CH.sub.3OH) for 5
minutes. After 5 minutes of etching the HDPE mesh was transferred
without washing or rinsing into 40 mL of an ammoniacal Ag(I) solution
containing 15.3 g/L AgNO.sub.3 and a stoichiometrically suitable
quantity of NH.sub.4OH (28 vol. %). The HDPE mesh was kept in this
solution for 2 minutes. The HDPE mesh was then transferred without
washing or rinsing into 150 mL of a solution containing 28 g/L NaOH.
The NaOH solution immediately became brown. After mixing for 2
minutes, the solution became clear and colorless and the mesh was tan
in color. When the agitation was stopped, the HDPE mesh was removed
from solution and washed with distilled water. After washing and
rinsing the mesh appeared to be tan in color as a consequence of the
coating with silver compounds.

The coated HDPE mesh was analyzed for silver content and for
antimicrobial activity against Pseudomonas Aeruginosa and
Staphylococcus Aureus. These samples contained between 0.04 and 0.08
mg/cm.sup.2 total silver. The bactericidal activities against
Pseudomonas Aeruginosa and Staphylococcus Aureus were positive. The
CZOI was estimated at about 10 mm.

Example 11

A patterned wound dressing made of a perforated plastic carrier
material with a skin adhesive layer comprised of a hydrophobic cross-
linked silicon gel (trade-mark Mepitel.TM., product of Molnlycke
Health Care AB, Sweden), dimensions 8.times.15 cm was exposed to a
solution containing 15 g/L NaOH at room temperature for 5 minutes.
Under conditions of agitation 40 mL of a solution containing 15.3 g/L
AgNO.sub.3 and a proper volume of NH.sub.4OH (28 vol. %) was added.
The wound dressing was kept in this solution and agitated for the next
5 minutes. The wound dressing was then removed from the solution and
carefully washed with distilled water. Drops of water were removed
with a soft paper and the wound dressing was dried at room
temperature.

The coated wound dressing was analyzed for antimicrobial activity
against Pseudomonas Aeruginosa and Staphylococcus Aureus. MH plates
and Tryptic Soy Broth were used for analysis. Pseudomonas Aeruginosa
standard was set to 0.5 McFarland standard. One hour of bactericidal
activity of the coated wound dressing against the bacteria where TSB
broths were incubated for 24 hours was positive. The controlled zones
of inhibition (CZOI), for the bacterial growth (bacteriostatic
activity) were above 8 mm. The same samples of coated wound dressing
were tested for seven days for antimicrobial activity. The values of
CZOI after 2 days were 20.5 mm, after 3 days 19 mm, after 4 days 20.5
mm, after 5 days 19 mm and after 7 days 7 mm. These results show very
good resistance towards bacteria for a relatively long time (7 days).

Example 12

A patterned wound dressing made of a perforated plastic carrier
material with a skin adhesive layer comprised of a hydrophobic cross-
linked silicon gel (trade-mark Mepitel.TM., product of Molnlycke
Health Care AB, Sweden), dimensions 8.times.15 cm was exposed to 500
mL of a 1% AgNO.sub.3 solution. To this solution was added 200 mL of a
solution containing 20 g K.sub.2S.sub.2O.sub.8 and mixing was
continuous for the next 20 minutes. The wound dressing was then
removed from the solution and carefully washed with distilled water.
Drops of water were removed with soft paper and the wound dressing was
dried at room temperature.

The coated wound dressing contained 0.25-0.55 mg/cm.sup.2 of total
silver. The coated wound dressing was then analyzed for antimicrobial
activity in the same manner as described in Example 11. The results
showed excellent antimicrobial activity for 7 days.

Example 13

A patterned wound dressing made of a perforated plastic carrier
material with a skin adhesive layer comprised of a hydrophobic cross-
linked silicon gel (trade-mark Mepitel.TM., product of Molnlycke
Health Care AB, Sweden), dimensions 8.times.15 cm was coated in a way
as described in Example 12, except that (NH.sub.4).sub.2 S.sub.2O.sub.
8 was used as an oxidizing agent instead of K.sub.2S.sub.2O.sub.8, in
the same amount and in the same manner as described in Example 12.

The coated wound dressing produced as described in this example was
analyzed for the antimicrobial activity. The results showed excellent
antimicrobial activity.

Example 14

A slurry was prepared by mixing 500 mL of a 1% AgNO.sub.3 solution and
200 mL of an aqueous solution containing 20 g K.sub.2S.sub.2O.sub.8
for 10 minutes. To this slurry a patterned wound dressing made of a
perforated plastic carrier material with a skin adhesive layer
comprised of a hydrophobic cross-linked silicon gel (trade-mark
Mepitel.TM., product of Molnlycke Health Care AB, Sweden), dimensions
of 8.times.15 cm was added and mixing was continued for the next 20
minutes. The coated wound dressing was then removed from the slurry,
carefully washed with water then dried as described in the Example 12.
The coated wound dressing was black-greyish in appearance.

The antimicrobial activity of the coated wound dressing was tested in
a way described in Example 11. The results showed excellent
antimicrobial activity for seven days.

Examples 15-16

All method steps were performed at room temperature (22 degrees
Celsius.+-.2 degrees. Celsius), unless otherwise specified.

Samples of HDPE mesh were coated with oxidized silver species as
follows. HDPE mesh with dimensions 10.times.10 cm were immersed into
100 mL of a 1% AgNO.sub.3 solution and thoroughly wetted. After the
exposure of the HDPE mesh to the solution for 10 minutes, 20 mL of a
solution containing either 250 g/L of (NH.sub.4).sub.2S.sub.2O.sub.8
or 250 g/L of K.sub.2S.sub.2O.sub.8 was added under magnetic stirring.
The mixing was continued for the next 15 minutes. The coated HDPE mesh
was then removed from the slurry and was observed to be grayish-black
in appearance. After coating, the HDPE mesh was washed with water and
then dried.

The bacteriostatic activity for the controlled zone of inhibition
(CZOI) of bacterial or fungal growth was tested against Pseudomonas
Aeruginosa, Staphylococcus Aureus or Candida Albicans, using standard
procedures as described in the literature.

Discussion of Examples 15-16

(a) Deposition of Silver Deposition Products Using (NH.sub.4).sub.
2S.sub.2O.sub.8

Upon addition of ammonium persulfate to the AgNO.sub.3 solution, a
gradual color change from colorless through yellow, brown and finally
to a cloud solution containing grayish-black precipitate was observed.
Time for the appearance of the grayish-black precipitate at room
temperature was estimated at 5 to 10 minutes. It was noted that if the
reaction takes place at temperatures above 30 degrees Celsius, the
precipitation and color change do not occur.

Persulfates are powerful oxidizing agents. In aqueous solutions
persulfates can be reduced to sulfates (S. I. Zhdanov, Sulfur,
Selenium, Tellurium and Polonium, in Standard Potentials in Aqueous
Solutions, A. J. Bard, R. Parsons and J. Jordan Editors, Marcel Dekker
Inc., New York (1985). A consequence of the reduction of persulfate is
the oxidation of Ag(I) to Ag(II) and Ag(II) to Ag(III). The grayish-
black precipitate deposited on the HDPE mesh was formed as a result of
the reduction of persulfate and a consequent oxidation of Ag(I) ions.

During precipitation of the deposition product, the pH of the solution
dropped from about 2 to below 1. The decrease in pH of the solution
was more significant when K.sub.2S.sub.2O.sub.8 is used as an
oxidizing agent instead of (NH.sub.4).sub.2S.sub.2O.sub.8, in that a
decrease in pH from about 7 to below 1 was observed.

(b) Properties of Deposition Products Produced Using (NH.sub.4).sub.
2S.sub.2O.sub.8

The grayish-black precipitate itself represents a mixture of silver
argentic nitrate Ag(Ag.sub.3O.sub.4).sub.2NO.sub.3.revreaction.Ag.sub.
7NO.sub.11 and Ag.sub.2SO.sub.4. Indeed, as found by XRD analysis, the
peaks in the patterns showed a reasonable match for Ag.sub.2SO.sub.4
and Ag.sub.7O.sub.8NO.sub.3 (FIG. 1). It is apparent that the
oxidation of AgNO.sub.3 with (NH.sub.4).sub.2S.sub.2O.sub.8 leads to
the precipitation of silver oxy-salt Ag.sub.7NO.sub.11 and also Ag.sub.
2SO.sub.4. The precipitation of Ag.sub.2SO.sub.4 is usually not
observed when K.sub.2S.sub.2O.sub.8 is used as an oxidizing agent of
Ag(I) ions (see the discussion below relating to oxidation with K.sub.
2S.sub.2O.sub.8).

FIG. 2 provides a SEM micrograph of the grayish black precipitate. The
smaller "cubical" particles represent Ag.sub.7O.sub.8NO.sub.3 and
their size, based on SEM is estimated at about 2.5 .mu.m. The shape of
these particles was found to be in very good agreement with the
results of Skanavi-Grigoreva (M. S. Skanavi-Grigoreva, I. L.
Shimanovich, Zh. Obsh., Khim., 24, 1490(1954)). who produced this
material by the electrolysis of an aqueous AgNO.sub.3 solution. The
larger, cylindrical particles represent silver sulfate (Ag.sub.2SO.sub.
4).

(c) Deposition of Silver Deposition Products Using K.sub.2S.sub.
2SO.sub.8

Some differences in the formation of the grayish-black precipitate
were observed when K.sub.2S.sub.2O.sub.8 was used instead of (NH.sub.
4).sub.2S.sub.2O.sub.8, as the oxidizing agent of Ag(I). The
precipitation of the grayish-black compound was significantly faster,
and occurred within 1 minute upon addition of K.sub.2S.sub.2O.sub.8 to
the AgNO.sub.3 solution. During this time, the pH of the solution
changed from the initial pH of about 7 to below 1 after the
precipitation.

(d) Properties of Deposition Products Produced Using K.sub.2S.sub.
2O.sub.8

As determined by XRD analysis in FIG. 3, all the peaks in the pattern
exactly match the compound of composition Ag.sub.7O.sub.8NO.sub.3. No
other compounds were identified in this XRD pattern.

The theoretical amount of Ag in the compound Ag.sub.7O.sub.8NO.sub.3
is 79.90%. The chemical analysis determined that the grayish black
precipitate contained about 78.80% Ag. This result shows a good
agreement of the experiments with the theory.

The SEM micrographs of the powder produced by the chemical oxidation
of AgNO.sub.3 with K.sub.2S.sub.2O.sub.8 are presented in FIG. 4. It
appears that the particles are uniform and cubical in their shape. The
size of these particles is estimated at about 2.5 .mu.m.

(e) Antimicrobial Activity

The comparison of the SEM micrographs of uncoated and coated HDPE mesh
samples is presented in FIG. 5. As shown in FIG. 5, the surface of the
HDPE is partially covered with the Ag(Ag.sub.3O.sub.4).sub.2NO.sub.3
particulates.

These samples were tested for bioactivity against the bacteria
Pseudomonas Aeruginosa, Staphylococcus Aureus or fungi Candida
Albicans. As can be seen from the photographs presented in FIG. 6,
clear zones surrounding the test samples (where a growth of tested
microorganisms did not occur) were observed in all cases for
Staphylococcus Aureus (a gram-positive bacteria), Pseudomonas
Aeuguginosa (a gram-negative bacteria) and Candida Albicans (an
example of fungi). The size of the controlled zone of inhibition
(CZOI), where the growth of tested microorganisms was not observed,
was estimated at 3 mm to 5 mm for all tested samples. These results
suggest that the deposition products have antibacterial and antifungal
properties. Furthermore these results are in agreement with previously
published results, where was suggested that only oxidized silver
species, but not metallic silver exhibit an antimicrobial activity.

(f) Conclusions Relating to Examples 15-16

It has been demonstrated that deposition products, namely those of
composition Ag.sub.7NO.sub.11.times.3Ag.sub.2SO.sub.4 or Ag.sub.
7NO.sub.11 can successfully be deposited as powders or on a substrate
such as HDPE mesh, by a simple reaction between AgNO.sub.3 and (NH.sub.
4)S.sub.2O.sub.8 or K.sub.2S.sub.2O.sub.8. These compounds are soluble
in both concentrated HNO.sub.3 or NH.sub.4OH.

Example 17

Samples of a substrate consisting of a patterned wound dressing made
of a perforated plastic carrier material with a skin adhesive layer
comprised of a hydrophobic cross-linked silicon gel (trade-mark
Mepitel.TM., product of Molnlycke Health Care AB, Sweden) were
subjected to SEM micrography to observe the density and coverage on
the substrate of a deposition product deposited on the substrate in
accordance with the second and third aspects of the invention, and to
XRD analysis to analyze the composition of the deposition product
deposited on the substrate.

FIG. 7 depicts an uncoated sample of the Mepitel.TM. wound dressing at
a magnification of 30.times.. FIGS. 8-11 depict samples of composite
materials which have been produced according to the second and third
aspects of the invention in the same manner as described in Example
14.

FIG. 8 depicts a composite material comprising a coated sample of the
Mepitel.TM. wound dressing at a magnification of 40.times., in which a
relatively low amount of deposition product has been deposited on the
substrate. FIG. 9 depicts the composite material of FIG. 8 at a
magnification of 2000.times., and clearly shows that the density and
coverage of the deposition product is such that the skin adhesive
layer of the Mepitel.TM. wound dressing is relatively unobstructed by
the deposition product.

FIG. 10 depicts a composite material comprising a coated sample of the
Mepitel.TM. wound dressing at a magnification of 40.times., in which a
higher amount of deposition product has been deposited on the
substrate in comparison with FIG. 8 and FIG. 9. FIG. 11 depicts the
composite material of FIG. 10 at a magnification of 2000.times., and
clearly shows that the skin adhesive layer remains relatively
unobstructed by the deposition product.

FIG. 12 depicts an XRD pattern for an uncoated sample of the
Mepitel.TM. wound dressing. FIG. 13 depicts an XRD pattern for a
composite material comprising a sample of the Mepitel.TM. wound
dressing which has been coated with a deposition product according to
the second and third aspects of the invention in the same manner as
described in Example 14. FIG. 14 superimposes the XRD patterns from
FIG. 12 and FIG. 13.

Referring to FIG. 14, the peaks which are observed in the pattern from
FIG. 13 but which are not observed in the pattern from FIG. 12 may be
attributed to the deposition product. These peaks define the
deposition product as comprising at least some amount of Ag.sub.7O.sub.
8NO.sub.3.

Example 18

Samples of a substrate consisting of a patterned wound dressing made
of a perforated plastic carrier material with a skin adhesive layer
comprised of a hydrophobic cross-linked silicon gel (trade-mark
Mepitel.TM., product of Molnlycke Health Care AB, Sweden) coated with
0.6 mg/cm.sup.2 of total silver according to the second and third
aspects of the invention in the same manner as described in Example 14
were exposed to a solution containing 10 g/L Na.sub.2S. After 10
minutes of exposure to the Na.sub.2S solution the coated wound
dressing samples were carefully washed with water until pH 7.

After drying, the samples were tested for antimicrobial activity
against Pseudomonas Aeruginosa and Staphylococcus Aureus using
standard procedures. Clear zones of inhibition of bacterial growth
surrounding test samples were observed for both Pseudomonas Aeruginosa
and Staphylococcus Aureus, suggesting that a deposition product
produced according to the second and third aspects of the invention
will exhibit an antimicrobial activity even after exposure to a
sulfide containing environment.



United States Patent 6,989,157
Gillis , et al. January 24, 2006
Dry powders of metal-containing compounds

Abstract

Dry powders of metal-containing compounds are disclosed. Methods of
preparing and using the dry powders, particularly in the treatment of
a subject having a condition, are also disclosed. The metal-containing
material can be, for example, an antimicrobial material, an
antibacterial material, an anti-inflammatory material, an anti-fungal
material, an anti-viral material, an anti-cancer material, a pro-
apoptosis material, and/or an MMP modulating material. In certain
embodiments, the metal-containing material is an atomically
disordered, silver-containing material.
Inventors: Gillis; Scott H. (Concord, MA), Schechter; Paul (Dover,
MA), Burrell; Robert E. (Alberta, CA)
Assignee: Nucryst Pharmaceuticals Corp. (Fort Saskatchewan, CA)
Appl. No.: 10/277,298
Filed: October 22, 2002
Related U.S. Patent Documents

Application Number Filing Date Patent Number Issue Date
10159587 May., 2002
10131568 Apr., 2002
10131511 Apr., 2002
10131509 Apr., 2002
10128208 Apr., 2002
09916757 Jul., 2001 6692773
09840637 Apr., 2001
09628735 Jul., 2000
60285884 Apr., 2001

Current U.S. Class: 424/618 ; 424/400; 424/489; 424/490; 424/617;
424/619; 424/646; 424/649; 514/492; 514/495; 514/849; 514/853;
514/888; 514/924; 514/951; 514/952; 514/958
Current International Class: A01N 59/16 (20060101)
Field of Search: 424/400,489,490,617-619,646,649
514/492,495,849,853,888,924,951,952,958
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Primary Examiner: Pak; John
Attorney, Agent or Firm: Fish & Richardson P.C.
Parent Case Text


CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part and claims the benefit of
priority under 35 U.S.C. .sctn.120 of: U.S. patent application Ser.
No. 09/628,735, filed Jul. 27, 2000, and entitled "Treatment of
Hyperproliferative Skin Disorders and Diseases" which is now
abandoned; U.S. patent application Ser. No. 09/916,757, filed Jul. 27,
2001, and entitled "Treatment of Hyperproliferative Skin Disorders and
Diseases," which is now U.S. Pat. No. 6,692,773; U.S. patent
application Ser. No. 09/840,637, filed Apr. 23, 2001, and entitled
"Treatment of Acne;" U.S. patent application Ser. No. 10/131,509,
filed Apr. 23, 2002, and entitled "Treatment of Mucosal Membranes;"
U.S. patent application Ser. No. 10/131,511, filed Apr. 23, 2002, and
entitled "Treatment of Inflammatory Skin Conditions;" U.S. patent
application Ser. No. 10/131,568, filed Apr. 23, 2002, and entitled
"Method of Induction of Apoptosis and Inhibition of Matrix
Metalloproteinases Using Antimicrobial Metals;" U.S. patent
application Ser. No. 10/159,587, filed May 30, 2002, and entitled
"Method of Induction of Apoptosis and Inhibition of Matrix
Metalloproteinases Using Antimicrobial Metals;" and U.S. patent
application Ser. No. 10/128,208, filed Apr. 23, 2002, and entitled
"Therapeutic Treatments Using the Direct Application of Antimicrobial
Metal Compositions," which claims priority from Provisional
Application Ser. No. 60/285,884, filed Apr. 23, 2001, and entitled
"Therapeutic Treatments Using the Direct Application of Noble Metal
Compositions." Each of these applications is incorporated by
reference.
Claims


What is claimed is:

1. A method of treating a human or animal subject having a condition,
the method comprising: contacting an area of the subject having the
condition with an antimicrobial nanocrystalline metal-containing
compound by dry inhaling a safe and therapeutically effective amount
of a free-standing powder of the antimicrobial nanocrystalline metal-
containing compound, wherein: the condition is selected from the group
consisting of bacterial respiratory conditions, microbial respiratory
conditions, fungal respiratory conditions and viral respiratory
conditions, the antimicrobial nanocrystalline metal-containing
compound contains a metal selected from the group consisting of
silver, gold, platinum and palladium, and the free-standing powder of
the antimicrobial nanocrystalline metal-containing compound has an
average particle size of less than about two microns.

2. The method of claim 1, wherein the antimicrobial nanocrystalline
metal-containing compound is selected from the group consisting of
silver, gold, platinum, palladium, and alloys thereof.

3. The method of claim 1, wherein the antimicrobial nanocrystalline
metal-containing compound is an oxide, nitride, boride, halide or
hydride of silver, gold, platinum or palladium.

4. The method of claim 1, wherein the antimicrobial nanocrystalline
metal-containing compound comprises silver.

5. The method of claim 1, antimicrobial nanocrystalline metal-
containing compound comprises an ionic compound of silver, gold,
platinum or palladium.

6. The method of claim 1, wherein the antimicrobial nanocrystalline
metal-containing compound comprises atoms, molecules or clusters of
silver, gold, platinum or palladium.

7. The method of claim 1, wherein the antimicrobial nanocrystalline
metal-containing compound comprises an atomically disordered,
crystalline compound of silver, gold, platinum or palladium.

8. The method of claim 1, wherein the condition is selected from the
group consisting of bronchitis, tuberculosis, pneumonia, sinusitis,
and pharyngitis.

9. The method of claim 1, wherein the free-standing powder is inhaled
with a dry powder inhaler.

10. The method of claim 1, wherein the free-standing powder has an
average particle size of about one micron or less.

11. The method of claim 1, wherein the condition is a bacterial
respiratory condition.

12. The method of claim 1, wherein the condition is a microbial
respiratory condition.

13. The method of claim 1, wherein the condition is a fungal
respiratory condition.

14. The method of claim 1, wherein the condition is a viral
respiratory condition.

15. The method of claim 1, wherein the condition is bronchitis.

16. The method of claim 1, wherein the condition is tuberculosis.

17. The method of claim 1, wherein the condition is pneumonia.

18. The method of claim 1, wherein the condition is sinusitis.

19. The method of claim 1, wherein the condition is pharyngitis.

20. The method of claim 1, wherein the condition is pneumonia and the
antimicrobial nanocrystalline metal-containing compound comprises
silver.
Description


TECHNICAL FIELD

The invention relates to dry powders of metal-containing compounds, as
well as their preparation and use, particularly in the treatment of a
subject having a condition.

BACKGROUND

It is generally desirable to treat a subject (e.g., a human) that has
an undesirable condition. Many different compositions have been
developed to treat undesirable conditions. For example, certain forms
of silver have been reported to be effective in treating some
undesirable skin conditions.

SUMMARY

The invention relates to dry powders of metal-containing compounds, as
well as their preparation and use, particularly in the treatment of a
subject having a condition.

In one aspect, the invention features a method of treating a subject
having a condition. The method includes contacting an area of the
subject having the condition with a nanocrystalline metal-containing
compound by injecting a free-standing powder of the nanocrystalline
metal-containing compound into the subject.

In another aspect, the invention features a method of treating a
subject having a condition. The method includes contacting an area of
the subject having the condition with a nanocrystalline metal-
containing compound by inhaling a free-standing powder of the
nanocrystalline metal-containing compound.

In a further aspect, the invention features a method of treating a
subject having a condition. The method includes contacting an area of
the subject having the condition with a nanocrystalline metal-
containing compound by inhaling a free-standing powder of the
nanocrystalline metal-containing compound.

Embodiments of the invention can include one or more of the following
features.

The metal-containing compound can be a metal or an alloy.

The metal-containing compound can be, for example, a metal oxide, a
metal nitride, a metal boride, a metal halide or a metal hydride.

The metal-containing compound can contain, for example, silver, gold,
platinum and/or palladium.

The metal-containing compound can be an ionic compound.

The metal-containing compound can be in the form of atoms, molecules
and/or clusters.

The metal-containing compound can be an antimicrobial compound.

The condition can be, for example, a bacterial condition, a microbial
condition, an inflammatory condition, a fungal condition, a viral
condition, an autoimmune condition, an idiopathic condition, a
noncancerous growth and/or a cancerous condition.

The condition can be, for example, a skin or integument condition, a
respiratory condition, a musculo-skeletal condition, a circulatory
condition, a mucosal or serosal condition and/or a cancerous
condition.

The free-standing powder can be injected with a needleless injector.

The free-standing powder can have an average particle size of about
two microns or less.

The methods can include monitoring a subject after contacting the
subject with the metal-containing material. For example, a subject can
be monitored at relatively regular intervals (e.g., about once an
hour, about once every eight hours, about once a day, about once a
week, about two times a month, about three times a month, about four
times a month).

Other features and advantages of the methods will be apparent from the
description and drawings, and from the claims.

DESCRIPTION OF DRAWINGS

FIG. 1 is a schematic view of a deposition system;

FIG. 2 is a graph showing the efficacy of different forms of silver on
erythema;

FIG. 3 is a graph showing the efficacy of different forms of silver on
edema;

FIG. 4 is a graph showing MMP activity of incision fluids recovered
from incisions dressed with materials;

FIG. 5 is a graph showing total protease activity of incision fluids
recovered from different dressings;

FIG. 6 is a graph showing the concentrations (ng/ml) of active MMP-9
in fluid samples recovered from ulcers dressed with different
materials;

FIG. 7 is a graph showing the concentrations (ng/ml) of active MMP-2
in fluid samples recovered from ulcers dressed with different
materials;

FIG. 8 is a graph showing the concentrations (pg/ml) of active
TNF-.alpha. in fluid samples recovered from ulcers dressed with
different materials; and

FIG. 9 is a graph showing the concentrations (pg/ml) of active
IL-1.beta. in fluid samples recovered from ulcers dressed with
different materials.

DETAILED DESCRIPTION

The inventors have discovered that certain metal-containing materials
(e.g., antimicrobial, atomically disordered, nanocrystalline silver-
containing materials) can be used to treat a subject with a condition
by contacting an area of the subject having the condition with the
metal-containing material. As explained below, the metal-containing
material can be in any of a variety of forms when delivered to a
subject, and the metal-containing material can be delivered to a
subject in a variety of ways. As also explained below, the metal-
containing material can be used to treat various subjects, conditions,
and condition locations.

Without wishing to be bound by theory, it is believed that the
therapeutic properties of the metal-containing materials may be
explained by one or more potential mechanisms. In one potential
mechanism, it is believed that the metal-containing material (e.g.,
antimicrobial, atomically disordered, nanocrystalline silver-
containing materials) forms one or more metastable, relatively high
level metal hydroxide species (e.g., Ag(OH).sub.4.sup.3-, Ag(OH).sub.
6.sup.3-) that either directly or indirectly (e.g., via the formation
of one or more biological mediators) provide the observed therapeutic
properties. In another potential mechanism, it is believed that the
metal-containing material is capable of releasing clusters of the
metal (e.g., clusters of Ag.sup.0, clusters of Ag.sup.1, clusters
containing both Ag.sup.+ and Ag.sup.0) that provide the observed
therapeutic properties. It is believed that combinations of potential
mechanisms may result in the observed therapeutic effect of the metal-
containing material.

In general, clusters refer to relatively small groups of atoms, ions
or the like. For example, a cluster can contain at least two (e.g., at
least three, at least four, at least five, at least six, at least
seven, at least eight, at least nine, at least 10, at least 11, at
least 12, at least 13, at least 14, at least 15, at least 20, at least
30, at least 40, at least 50, at least 60, at least 70, at least 80,
at least 90) atoms, ions or the like, and/or at most 1,000 (e.g., at
most 900, at most 800, at most 700, at most 600, at most 500, at most
400, at most 300, at most 200, at most 100) atoms, ions or the like.
Clusters are described, for example, in R. P. Andres et al., "Research
Opportunities on Cluster and Cluster-Assembled Materials", J. Mater.
Res. Vol. 4, No 3, 1989, p. 704. In certain embodiments, a cluster
(e.g., a cluster containing silver) can contain less than the 14 atoms
and have a normal face centered cubic crystal lattice.

Materials

The metal-containing material can be an ionic material or a non-ionic
material. The metal-containing material can be, for example, an atom,
a molecule, or a cluster.

In general, the metal-containing material is a metal or an alloy.
Examples of metals that can be contained in metal-containing materials
include Group I A metals, Group II A metals, Group III A metals, Group
IV A metals, Group V A metals, Group VI A metals, Group VII A metals,
Group VIII A metals, Group I B metals, Group II B metals, members of
the lanthanide metal series, and members of the actinide metal series.
In certain embodiments, metal-containing materials contain silver,
gold, platinum, palladium, iridium, zinc, copper, tin, antimony, and/
or bismuth. Examples of silver-containing metals include colloidal
silver, silver nitrate and silver sulfadiazine.

In addition to one or more metals, a metal-containing material can
contain oxygen, nitrogen, carbon, boron, sulfur, a halogen (e.g.,
fluorine, chlorine, bromine, iodine) and/or hydrogen. Examples of such
metal-containing materials include metal oxides, metal nitrides, metal
carbides, metal borides, metal sulfides, metal halides (e.g., metal
fluorides, metal chlorides, metal bromides, metal iodides) and metal
hydrides. In certain embodiments, a metal-containing material contains
at least about one atomic percent (e.g., at least about three atomic
percent, at least about five atomic percent) and/or at most about 20
atomic percent (e.g., at most about 15 atomic percent, at most about
12 atomic percent, at most about 10 atomic percent) of nonmetallic
elements. For example, in some embodiments, a silver-containing
material can contain oxygen in an amount from about five atomic
percent to about 20 atomic percent (e.g., from about five atomic
percent to about 15 atomic percent, from about eight atomic percent to
about 12 atomic percent).

In certain embodiments, the metal-containing materials are an
antimicrobial material, an atomically disordered crystalline material,
and/or a nanocrystalline material.

As used herein, an antimicrobial material herein refers to a material
that has sufficient antimicrobial activity to have a beneficial
therapeutic effect. In certain embodiments, an antimicrobial material
has a corrected zone of inhibition ("CZOI") of at least about two
millimeters (e.g., at least about three millimeters, at least about
four millimeters, at lest about five millimeters, at least about six
millimeters, at least about seven millimeters, at least about eight
millimeters, at least about nine millimeters, at least about 10
millimeters). The CZOI of a material is determined as follows. The
material is formed as a coating on a dressing (see discussion below).
Basal medium Eagle (BME) with Earle's salts and L-glutamine is
modified with calf/serum (10%) and 1.5% agar prior to being dispensed
(15 ml) into Petri dishes. The agar containing Petri dishes are
allowed to surface dry prior to being inoculated with a lawn of
Staphylococcus aureus ATCC #25923. The inoculant is prepared from
Bactrol Discs (Difco, M.) which are reconstituted as per the
manufacturer's directions. Immediately after inoculation, the coatings
to be tested are placed on the surface of the agar. The dishes are
incubated for 24 hours at 37.degree. C. After this incubation period,
the zone of inhibition ("ZOI") is measured and the CZOI is calculated
as the ZOI minus the diameter of the test material in contact with the
agar. It is to be noted that, while this test for antimicrobial
properties is performed on materials that are in the form of a coating
on a substrate (e.g., in the form of a dressing), antimicrobial
materials are not limited to materials that are coated on a substrate.
Rather, a material in any form may be antimicrobial, but it is in the
form of a coating on a substrate (e.g., in the form of a dressing)
when its antimicrobial properties are tested according to the
procedure described herein.

As referred to herein, an atomically disordered, crystalline material
(e.g., an atomically disordered, nanocrystalline material) means a
material that has more long range ordered, crystalline structure (a
lesser degree of defects) than the material has in a fully amorphous
state, but that also has less long range, ordered crystalline
structure (a higher degree of defects) than the material has in a bulk
crystalline state, such as in the form of a cast, wrought or plated
material. Examples of defects include point defects, vacancies, line
defects, grain boundaries, subgrain boundaries and amorphous regions.
Point defects are defects on a size scale of no more than about four
atomic spacings. A vacancy is the omission of an atom from its regular
atomic site in the crystal lattice. Line defects are defective regions
(e.g., edge dislocations, screw dislocations) that result in lattice
distortions along a line (which may or may not be a straight line),
and generally have a longer scale than point defects. In an edge
dislocation, a lattice displacement is produced by a plane of atoms
that forms a terminus of the lattice. In a screw dislocation, part of
the lattice is displaced with respect to an adjacent part of the
lattice. Grain boundaries separate regions having different
crystallographic orientation or misorientation (e.g., high angle grain
boundaries, low angle grain boundaries, including tilt boundaries and
twist boundaries). Subgrain boundaries refer to low angle grain
boundaries. An amorphous region is a region that does not exhibit long
range, ordered crystalline structure. In certain embodiments, an
atomically disordered, crystalline material (e.g., an atomically
disordered, nanocrystalline material) has a degree of atomic disorder
that is about the same as the degree of atomic disorder of the
nanocrystalline silver coating of a member of the Acticoat.RTM. family
of dressings (Smith & Nephew, Hull, UK) (e.g., an Acticoat.RTM.
dressing, an Acticoat7.RTM. dressing, an Acticoat.RTM. moisture
coating dressing, an Acticoat.RTM. absorbent dressings). In some
embodiments, an atomically disordered, crystalline material (e.g., an
atomically disordered, nanocrystalline material) has a degree of
atomic disorder that is about the same as the degree of atomic
disorder of the nanocrystalline silver coatings having a CZOI of at
least five millimeters that are disclosed in the examples of Burrell
et al., U.S. Pat. No. 5,958,440. In certain embodiments, an atomically
disordered, crystalline material (e.g., an atomically disordered,
nanocrystalline material), when contacted with an alcohol or water-
based electrolyte, is released into the alcohol or water-based
electrolyte (e.g., as ions, atoms, molecules and/or clusters) over a
time scale of at least about one hour (e.g., at least about two hours,
at least about 10 hours, at least about a day). Examples of alcohols
and/or water-based electrolytes include body fluids (e.g., blood,
urine, saliva) and body tissue (e.g., skin, muscle, bone).

As referred to herein, a nanocrystalline material is a single-phase
polycrystal or a multi-phase polycrystal having a maximum dimension of
about 100 nanometers or less (e.g., about 90 nanometers or less, about
80 nanometers or less, about 70 nanometers or less, about 60
nanometers or less, about 50 nanometers or less, about 40 nanometers
or less, about 30 nanometers or less, about 25 nanometers or less) in
at least one dimension.

Examples of antimicrobial metal-containing materials (which may or may
not also be an atomically disordered crystalline material or a
nanocrystalline material) include antimicrobial silver-containing
materials (e.g., antimicrobial silver, antimicrobial silver alloys,
antimicrobial silver oxides, antimicrobial silver carbides,
antimicrobial silver nitrides, antimicrobial silver borides,
antimicrobial silver sulfides, antimicrobial silver halides,
antimicrobial silver hydrides), antimicrobial gold-containing
materials (e.g., antimicrobial gold, antimicrobial gold alloys,
antimicrobial gold oxides, antimicrobial gold carbides, antimicrobial
gold nitrides, antimicrobial gold borides, antimicrobial gold
sulfides, antimicrobial gold halides, antimicrobial gold hydrides),
antimicrobial platinum-containing materials (e.g., antimicrobial
platinum, antimicrobial platinum alloys, antimicrobial platinum
oxides, antimicrobial platinum carbides, antimicrobial platinum
nitrides, antimicrobial platinum borides, antimicrobial platinum
sulfides, antimicrobial platinum halides, antimicrobial platinum
hydrides), antimicrobial palladium-containing materials (e.g.,
antimicrobial palladium, antimicrobial palladium alloys, antimicrobial
palladium oxides, antimicrobial palladium carbides, antimicrobial
palladium nitrides, antimicrobial palladium borides, antimicrobial
palladium sulfides, antimicrobial palladium halides, antimicrobial
palladium hydrides), antimicrobial iridium-containing materials (e.g.,
antimicrobial iridium, antimicrobial iridium alloys, antimicrobial
iridium oxides, antimicrobial iridium carbides, antimicrobial iridium
nitrides, antimicrobial iridium borides, antimicrobial iridium
sulfides, antimicrobial iridium halides, antimicrobial iridium
hydrides), antimicrobial zinc-containing materials (e.g.,
antimicrobial zinc, antimicrobial zinc alloys, antimicrobial zinc
oxides, antimicrobial zinc carbides, antimicrobial zinc nitrides,
antimicrobial zinc borides, antimicrobial zinc sulfides, antimicrobial
zinc halides, antimicrobial zinc hydrides), antimicrobial copper-
containing materials (e.g., antimicrobial copper, antimicrobial copper
alloys, antimicrobial copper oxides, antimicrobial copper carbides,
antimicrobial copper nitrides, antimicrobial copper borides,
antimicrobial copper sulfides, antimicrobial copper halides,
antimicrobial copper hydrides), antimicrobial tin-containing materials
(e.g., antimicrobial tin, antimicrobial tin alloys, antimicrobial tin
oxides, antimicrobial tin carbides, antimicrobial tin nitrides,
antimicrobial tin borides, antimicrobial tin sulfides, antimicrobial
tin halides, antimicrobial tin hydrides), antimicrobial antimony-
containing materials (e.g., antimicrobial antimony, antimicrobial
antimony alloys, antimicrobial antimony oxides, antimicrobial antimony
carbides, antimicrobial antimony nitrides, antimicrobial antimony
borides, antimicrobial antimony sulfides, antimicrobial antimony
halides, antimicrobial antimony hydrides), antimicrobial bismuth
containing materials (e.g., antimicrobial bismuth, antimicrobial
bismuth alloys, antimicrobial bismuth oxides, antimicrobial bismuth
carbides, antimicrobial bismuth nitrides, antimicrobial bismuth
borides, antimicrobial bismuth sulfides, antimicrobial bismuth
halides, antimicrobial antimony hydrides).

Examples of nanocrystalline metal-containing materials (which may or
may not also be an antimicrobial material or an atomically disordered
crystalline material) include nanocrystalline silver-containing
materials (e.g., nanocrystalline silver, nanocrystalline silver
alloys, nanocrystalline silver oxides, nanocrystalline silver
carbides, nanocrystalline silver nitrides, nanocrystalline silver
borides, nanocrystalline silver sulfides, nanocrystalline silver
halides, nanocrystalline silver hydrides), nanocrystalline gold-
containing materials (e.g., nanocrystalline gold, nanocrystalline gold
alloys, nanocrystalline gold oxides, nanocrystalline gold carbides,
nanocrystalline gold nitrides, nanocrystalline gold borides,
nanocrystalline gold sulfides, nanocrystalline gold halides,
nanocrystalline gold hydrides), nanocrystalline platinum-containing
materials (e.g., nanocrystalline platinum, nanocrystalline platinum
alloys, nanocrystalline platinum oxides, nanocrystalline platinum
carbides, nanocrystalline platinum nitrides, nanocrystalline platinum
borides, nanocrystalline platinum sulfides, nanocrystalline platinum
halides, nanocrystalline platinum hydrides), nanocrystalline palladium-
containing materials (e.g., nanocrystalline palladium, nanocrystalline
palladium alloys, nanocrystalline palladium oxides, nanocrystalline
palladium carbides, nanocrystalline palladium nitrides,
nanocrystalline palladium borides, nanocrystalline palladium sulfides,
nanocrystalline palladium halides, nanocrystalline palladium
hydrides), nanocrystalline iridium-containing materials (e.g.,
nanocrystalline iridium, nanocrystalline iridium alloys,
nanocrystalline iridium oxides, nanocrystalline iridium carbides,
nanocrystalline iridium nitrides, nanocrystalline iridium borides,
nanocrystalline iridium sulfides, nanocrystalline iridium halides,
nanocrystalline iridium hydrides), nanocrystalline zinc-containing
materials (e.g., nanocrystalline zinc, nanocrystalline zinc alloys,
nanocrystalline zinc oxides, nanocrystalline zinc carbides,
nanocrystalline zinc nitrides, nanocrystalline zinc borides,
nanocrystalline zinc sulfides, nanocrystalline zinc halides,
nanocrystalline zinc hydrides), nanocrystalline copper -containing
materials (e.g., nanocrystalline copper, nanocrystalline copper
alloys, nanocrystalline copper oxides, nanocrystalline copper
carbides, nanocrystalline copper nitrides, nanocrystalline copper
borides, nanocrystalline copper sulfides, nanocrystalline copper
halides, nanocrystalline copper hydrides), nanocrystalline tin-
containing materials (e.g., nanocrystalline tin, nanocrystalline tin
alloys, nanocrystalline tin oxides, nanocrystalline tin carbides,
nanocrystalline tin nitrides, nanocrystalline tin borides,
nanocrystalline tin sulfides, nanocrystalline tin halides,
nanocrystalline tin hydrides), nanocrystalline antimony-containing
materials (e.g., nanocrystalline antimony, nanocrystalline antimony
alloys, nanocrystalline antimony oxides, nanocrystalline antimony
carbides, nanocrystalline antimony nitrides, nanocrystalline antimony
borides, nanocrystalline antimony sulfides, nanocrystalline antimony
halides, nanocrystalline antimony hydrides), nanocrystalline bismuth
containing materials (e.g., nanocrystalline bismuth, nanocrystalline
bismuth alloys, nanocrystalline bismuth oxides, nanocrystalline
bismuth carbides, nanocrystalline bismuth nitrides, nanocrystalline
bismuth borides, nanocrystalline bismuth sulfides, nanocrystalline
bismuth halides, nanocrystalline antimony hydrides).

Examples of atomically disordered, crystalline metal-containing
material (which may or may not also be an antimicrobial material or a
nanocrystalline material) include atomically disordered, crystalline
silver-containing materials (e.g., atomically disordered, crystalline
silver; atomically disordered, crystalline silver alloys; atomically
disordered, crystalline silver oxides; atomically disordered,
crystalline silver carbides; atomically disordered, crystalline silver
nitrides; atomically disordered, crystalline silver borides;
atomically disordered, crystalline silver sulfides; atomically
disordered, crystalline silver halides; atomically disordered,
crystalline silver hydrides), atomically disordered, crystalline gold-
containing materials (atomically disordered, crystalline gold;
atomically disordered, crystalline gold alloys; atomically disordered,
crystalline gold oxides; atomically disordered, crystalline gold
carbides; atomically disordered, crystalline gold nitrides; atomically
disordered, crystalline gold borides; atomically disordered,
crystalline gold sulfides; atomically disordered, crystalline gold
halides; atomically disordered, crystalline gold hydrides), atomically
disordered, crystalline platinum-containing materials (e.g.,
atomically disordered, crystalline platinum; atomically disordered,
crystalline platinum alloys; atomically disordered, crystalline
platinum oxides; atomically disordered, crystalline platinum carbides;
atomically disordered, crystalline platinum nitrides; atomically
disordered, crystalline platinum borides; atomically disordered,
crystalline platinum sulfides; atomically disordered, crystalline
platinum halides; atomically disordered, crystalline platinum
hydrides), atomically disordered, crystalline palladium-containing
materials (e.g., atomically disordered, crystalline palladium;
atomically disordered, crystalline palladium alloys; atomically
disordered, crystalline palladium oxides; atomically disordered,
crystalline palladium carbides; atomically disordered, crystalline
palladium nitrides; atomically disordered, crystalline palladium
borides; atomically disordered, crystalline palladium sulfides;
atomically disordered, crystalline palladium halides; atomically
disordered, crystalline palladium hydrides), atomically disordered,
crystalline iridium-containing materials (e.g., atomically disordered,
crystalline iridium; atomically disordered, crystalline iridium
alloys; atomically disordered, crystalline iridium oxides; atomically
disordered, crystalline iridium carbides; atomically disordered,
crystalline iridium nitrides; atomically disordered, crystalline
iridium borides; atomically disordered, crystalline iridium sulfides;
atomically disordered, crystalline iridium halides; atomically
disordered, crystalline iridium hydrides), atomically disordered,
crystalline zinc-containing materials (e.g., atomically disordered,
crystalline zinc; atomically disordered, crystalline zinc alloys;
atomically disordered, crystalline zinc oxides; atomically disordered,
crystalline zinc carbides; atomically disordered, crystalline zinc
nitrides; atomically disordered, crystalline zinc borides; atomically
disordered, crystalline zinc sulfides; atomically disordered,
crystalline zinc halides; atomically disordered, crystalline zinc
hydrides), atomically disordered, crystalline copper-containing
materials (e.g., atomically disordered, crystalline copper; atomically
disordered, crystalline copper alloys; atomically disordered,
crystalline copper oxides; atomically disordered, crystalline copper
carbides; atomically disordered, crystalline copper nitrides;
atomically disordered, crystalline copper borides; atomically
disordered, crystalline copper sulfides; atomically disordered,
crystalline copper halides; atomically disordered, crystalline copper
hydrides), atomically disordered, crystalline tin-containing materials
(e.g., atomically disordered, crystalline tin; atomically disordered,
crystalline tin alloys; atomically disordered, crystalline tin oxides;
atomically disordered, crystalline tin carbides; atomically
disordered, crystalline tin nitrides; atomically disordered,
crystalline tin borides; atomically disordered, crystalline tin
sulfides; atomically disordered, crystalline tin halides; atomically
disordered, crystalline tin hydrides), atomically disordered,
crystalline antimony-containing materials (e.g., atomically
disordered, crystalline antimony; atomically disordered, crystalline
antimony alloys; atomically disordered, crystalline antimony oxides;
atomically disordered, crystalline antimony carbides; atomically
disordered, crystalline antimony nitrides; atomically disordered,
crystalline antimony borides; atomically disordered, crystalline
antimony sulfides; atomically disordered, crystalline antimony
halides; atomically disordered, crystalline antimony hydrides),
atomically disordered, crystalline bismuth-containing materials (e.g.,
atomically disordered, crystalline bismuth; atomically disordered,
crystalline bismuth alloys; atomically disordered, crystalline bismuth
oxides; atomically disordered, crystalline bismuth carbides;
atomically disordered, crystalline bismuth nitrides; atomically
disordered, crystalline bismuth borides; atomically disordered,
crystalline bismuth sulfides; atomically disordered, crystalline
bismuth halides; atomically disordered, crystalline bismuth hydrides).

Subjects

The metal-containing material can be used to treat, for example a
human or an animal (e.g., a dog, a cat, a horse, a bird, a reptile, an
amphibian, a fish, a turtle, a guinea pig, a hamster, a rodent, a cow,
a pig, a goat, a primate, a monkey, a chicken, a turkey, a buffalo, an
ostrich, a sheep, a llama).

Conditions and Condition Locations

The conditions that can be treated with the metal-containing material
include, for example, bacterial conditions, microbial conditions,
inflammatory conditions, fungal conditions, viral conditions,
autoimmune conditions, idiopathic conditions, noncancerous growths and/
or cancerous conditions (e.g., tumorous conditions, hematologic
malignancies). Such conditions can be associated with, for example,
one or more prions, parasites, fungi, viruses and/or bacteria. In
general, the location of the condition to be treated corresponds to
the type of condition to be treated.

In some embodiments, the condition can be a skin condition or a
integument condition (e.g., a microbial skin condition, an
inflammatory skin condition, a fungal skin condition, a viral skin
condition, an autoimmune skin condition, an idiopathic skin condition,
a cancerous skin condition, a microbial integument condition, an
inflammatory integument condition, a fungal integument condition, a
viral integument condition, an autoimmune integument condition, an
idiopathic integument condition, a cancerous integument condition).
Examples of skin conditions or integument conditions include burns,
eczema (e.g., atopic eczema, acrodermatitis continua, contact allergic
dermatitis, contact irritant dermatitis, dyshodrotic eczema,
pompholyx, lichen simplex chronicus, nummular eczema, seborrheic
dermatitis, stasis eczema), erythroderma, insect bites, mycosis
fungoides, pyoderma gangrenosum, eythrema multiforme, rosacea,
onychomyocosis, acne (e.g., acne vulgaris, neonatal acne, infantile
acne, pomade acne), psoriasis, Reiter's syndrome, pityriasis rubra
pilaris, hyperpigmentation, vitiligo, hypertropic scarring, keloids,
lichen plainus, age-related skin disorders (e.g., wrinkles, cellulite)
and hyperproliferative variants of the disorders of keratinization
(e.g., actinic keratosis, senile keratosis). Generally, the treatment
of skin or integument conditions involves contacting the metal-
containing material with the area of the skin having the condition. As
an example, a skin or integument condition can be treated by
contacting the area of skin having the condition with a dressing
having a coating of the metal-containing material. As another example,
a skin or integument condition can be treated by contacting the area
of skin having the condition with a solution containing the metal-
containing material. As an additional example, a skin or integument
condition can be treated by contacting the area of skin having the
condition with a pharmaceutical carrier composition containing the
metal-containing material. In the case of onychomycosis, the material
may be applied to the nail in an appropriate form (see below) such
that the material penetrates the hard nail to contact the affected
area.

In certain embodiments, the condition can be a respiratory condition
(e.g., a microbial respiratory condition, an inflammatory respiratory
condition, a fungal respiratory condition, a viral respiratory
condition, an autoimmune respiratory condition, an idiopathic
respiratory condition, a cancerous respiratory condition). Examples of
respiratory conditions include asthma, emphysema, bronchitis,
pulmonary edema, acute respiratory distress syndrome, bronchopulmonary
dysplasia, pulmonary fibrosis, pulmonary atelectasis, tuberculosis,
pneumonia, sinusitis, pharyngitis, mucositis, stomatitis, chronic
obstructive pulmonary disease, bronchiectasis, lupus pneumonitis and
cystic fibrosis. In general, the treatment of respiratory conditions
involves contacting the metal-containing material with the area of the
respiratory system having the condition. Areas of the respiratory
system include, for example, the oral cavity, the nasal cavity, and
the lungs. As an example, certain respiratory conditions can be
treated by inhaling a free standing powder of the metal-containing
material (e.g., with a dry powder inhaler). As another example,
certain respiratory conditions can be treated by inhaling an aerosol
containing the metal-containing material (e.g., with an inhaler).

In some embodiments, the condition can be a musculo-skeletal condition
(e.g., a microbial musculo-skeletal condition, an inflammatory musculo-
skeletal condition, a fungal musculo-skeletal condition, a viral
musculo-skeletal condition, an autoimmune musculo-skeletal condition,
an idiopathic musculo-skeletal condition, a cancerous musculo-skeletal
condition). A musculo-skeletal condition can be, for example, a
degenerative musculo-skeletal condition (e.g., arthritis) or a
traumatic musculo-skeletal condition (e.g., a torn or damaged muscle).
Examples of musculo-skeletal conditions include tendonitis,
osteomyclitis, fibromyalgia, bursitis and arthritis. Generally, the
treatment of musculo-skeletal conditions involves contacting the metal-
containing material compound with the area of the musculo-skeletal
system having the condition. Areas of the musculo-skeletal system
include, for example, the joints, the muscles, and the tendons. As an
example, certain musculo-skeletal conditions can be treated by
injecting (e.g., via a small needle injector) a solution containing
the metal-containing material into the subject. As another example,
certain musculo-skeletal conditions can be treated by injecting (e.g.,
via a needleless injector) a free standing powder of the metal-
containing material into the subject. As an additional example,
certain musculo-skeletal conditions can be treated by using a
pharmaceutical carrier composition of the metal-containing material,
such as a penetrating pharmaceutical carrier composition of the metal-
containing material (e.g., a composition containing DMSO).

In certain embodiments, the condition can be a circulatory condition
(e.g., a microbial circulatory condition, an inflammatory circulatory
condition, a fungal circulatory condition, a viral circulatory
condition, an autoimmune circulatory condition, an idiopathic
circulatory condition, a cancerous circulatory condition). As referred
to herein, circulatory conditions include lymphatic conditions.
Examples of circulatory conditions include arteriosclerosis,
septicemia, leukemia, ischemic vascular disease, lymphangitis and
atherosclerosis. In general, the treatment of circulatory conditions
involves contacting the metal-containing material with the area of the
circulatory system having the condition. Areas of the circulatory
system include, for example, the heart, the lymphatic system, blood,
blood vessels (e.g., arteries, veins). As an example, certain
circulatory conditions can be treated by injecting (e.g., via a small
needle injector) a solution containing the metal-containing material
into the subject. As another example, certain circulatory conditions
can be treated by injecting (e.g., via a needleless injector) a free
standing powder of the metal-containing material into the subject.

In some embodiments, the condition can be a mucosal or serosal
condition (e.g., a microbial mucosal or serosal condition, an
inflammatory mucosal or serosal condition, a fungal mucosal or serosal
condition, a viral mucosal or serosal condition, an autoimmune mucosal
or serosal condition, an idiopathic mucosal or serosal condition, a
cancerous mucosal or serosal condition). Examples of mucosal or
serosal conditions include pericarditis, Bowen's disease, stomatitis,
prostatitis, sinusitis, digestive disorders, esophageal ulcer, gastric
ulcer, duodenal ulcer, espohagitis, gastritis, enteritis,
enterogastric intestinal hemorrhage, toxic epidermal necrolysis
syndrome, Stevens Johnson syndrome, cystic fibrosis, bronchitis,
pneumonia (e.g., nosocomial pneumonia, ventilator-assisted pneumonia),
pharyngitis, common cold, ear infections, sore throat, sexually
transmitted diseases (e.g., syphilis, gonorrhea, herpes, genital
warts, HIV, chlamydia), inflammatory bowel disease, colitis,
hemorrhoids, thrush, dental conditions, oral conditions,
conjunctivitis, and periodontal conditions. Generally, the treatment
of mucosal or serosal conditions involves contacting the metal-
containing material with the area of a mucosal or serosal region
having the condition. Mucosal or serosal areas include, for example,
the oral cavity, the nasal cavity, the colon, the small intestine, the
large intestine, the stomach, and the esophagus. As an example,
certain mucosal or serosal conditions can be treated by inhaling a
free standing powder of the metal-containing material (e.g., with a
dry powder inhaler). As another example, certain mucosal or serosal
conditions can be treated by inhaling an aerosol containing the metal-
containing material (e.g., with an inhaler). As an additional example,
certain mucosal or serosal conditions can be treated by gargling or
spraying a solution of the metal-containing material.

In embodiments in which the metal-containing material is used to treat
hyperproliferation of cell growth (e.g., cancerous conditions, such as
malignant tumors, or non-cancerous conditions, such as benign tumors),
the metal-containing material can be used to induce apoptosis
(programmed cell death), modulate matrix metalloproteinases (MMPs) and/
or modulates cytokines by contacting affected tissue (e.g., a
hyperplastic tissue, a tumor tissue or a cancerous lesion) with the
metal-containing material. It has been observed that the metal-
containing material (e.g., an antimicrobial, atomically disordered,
silver-containing material) can be effective in preventing production
of a high number of MMPs and/or cytokines by certain cells without
necessarily reducing MMP and/or cytokine production by the same cells
to about zero. It is believed, however, that in certain embodiments,
the metal-containing material can be used to inhibit MMP and/or
cytokine production (e.g., bring MMP and/or cytokine production to
normal levels, desired levels, and/or about zero) in certain cells.

MMPs refer to any protease of the family of MMPs which are involved in
the degradation of connective tissues, such as collagen, elastins,
fibronectin, laminin, and other components of the extracellular
matrix, and associated with conditions in which excessive degradation
of extracellular matrix occurs, such as tumor invasion and metastasis.
Examples of MMPs include MMP-2 (secreted by fibroblasts and a wide
variety of other cell types) and MMP-9 (released by mononuclear
phagocytes, neutrophils, corneal epithelial cells, tumor cells,
cytotrophoblasts and keratinocytes). Cytokine refers to a
nonimmunoglobulin polypeptide secreted by monocytes and lymphocytes in
response to interaction with a specific antigen, a nonspecific
antigen, or a nonspecific soluble stimulus (e.g., endotoxin, other
cytokines). Cytokines affect the magnitude of inflammatory or immune
responses. Cytokines can be divided into several groups, which include
interferons, tumor necrosis factor (TNF), interleukins (IL-1 to IL-8),
transforming growth factors, and the hematopoietic colony-stimulating
factors. An example of a cytokine is TNF-.alpha.. A fibroblast is an
area connective tissue cell which is a flat-elongated cell with
cytoplasmic processes at each end having a flat, oval vesicular
nucleus. Fibroblasts which differentiate into chondroblasts,
collagenoblasts, and osteoblasts form the fibrous tissues in the body,
tendons, aponeuroses, supporting and binding tissues of all sorts.
Hyperplastic tissue refers to tissue in which there is an abnormal
multiplication or increase in the number of cells in a normal
arrangement in normal tissue or an organ. A tumor refers to
spontaneous growth of tissue in which multiplication of cells is
abnormal, uncontrolled and progressive. A tumor generally serves no
useful function and grows at the expense of the healthy organism. A
cancerous lesion is a tumor of epithelial tissue, or malignant, new
growth made up of epithelial cells tending to infiltrate surrounding
tissues and to give rise to metastases. As used in reference to the
skin, a cancerous lesion means a lesion which may be a result of a
primary cancer, or a metastasis to the site from a local tumor or from
a tumor in a distant site. It may take the form of a cavity, an open
area on the surface of the skin, skin nodules, or a nodular growth
extending from the surface of the skin.

Conditions characterized by undesirable MMP activity include ulcers,
asthma, acute respiratory distress syndrome, skin disorders, skin
aging, keratoconus, restenosis, osteo- and rheumatoid arthritis,
degenerative joint disease, bone disease, wounds, cancer including
cell proliferation, invasiveness, metastasis (carcinoma, fibrosarcoma,
osteosarcoma), hypovolemic shock, periodontal disease, epidermolysis
bullosa, scleritis, atherosclerosis, multiple sclerosis, inflammatory
diseases of the central nervous system, vascular leakage syndrome,
collagenase induced disease, adhesions of the peritoneum, strictures
of the esophagus or bowel, ureteral or urethral strictures, and
biliary strictures. Excessive TNF production has been reported in
diseases which are characterized by excessive MMP activity, such as
autoimmune disease, cancer, cachexia, HIV infection, and
cardiovascular conditions.

Forms of the Material and Methods of Applying the Material

In general, the metal-containing material can be in any desired form
or formulation. For example, the material can be a coating on a
substrate (e.g., in the form of a dressing), a free standing powder, a
solution, or disposed within a pharmaceutically acceptable carrier.

In some embodiments, the metal-containing material can act as a
preservative. In such embodiments, a form or formulation containing
the metal-containing material can be prepared without additional
preservatives. Moreover, in embodiments in which the metal-containing
material acts as a preservative, the metal-containing material may be
included in a therapeutic formulation containing other therapeutic
agents (e.g., the metal-containing material may be included primarily
in certain therapeutic compositions to act as a preservative).

Moreover, the material can be applied to the subject in any of a
variety of ways, generally depending upon the form of the material as
applied and/or the location of the condition to be treated. In
general, the amount of material used is selected so that the desired
therapeutic effect (e.g., reduction in the condition being treated) is
achieved while the material introduces an acceptable level of toxicity
(e.g., little or no toxicity) to the subject. Generally, the amount of
the material used will vary with the conditions being treated, the
stage of advancement of the condition, the age and type of host, and
the type, concentration and form of the material as applied.
Appropriate amounts in any given instance will be readily apparent to
those skilled in the art or capable of determination by routine
experimentation. In some embodiments, a single application of the
material may be sufficient. In certain embodiments, the material may
be applied repeatedly over a period of time, such as several times a
day for a period of days or weeks.

Substrate Coatings

Examples of commercially available metal-containing materials include
the Acticoat.RTM. family of dressings (Smith & Nephew, Hull, UK),
which are formed of antimicrobial, atomically disordered,
nanocrystalline silver-containing material coated on one or more
substrates. Such dressings include the Acticoat.RTM. dressings, the
Acticoat7.RTM. dressings, the Acticoat.RTM. moisture coating
dressings, and the Acticoat.RTM. absorbent dressings.

A coating of a metal-containing material (e.g., an antimicrobial,
atomically disordered, nanocrystalline silver-containing material) can
be formed on a substrate using a desired technique. In certain
embodiments, the coating is formed by depositing the material on the
substrate surface using chemical vapor deposition, physical vapor
deposition, and/or liquid phase deposition. Exemplary deposition
methods include vacuum evaporation deposition, arc evaporation
deposition, sputter deposition, magnetron sputter deposition and ion
plating.

In some embodiments, the coating is prepared using physical vapor
deposition. FIG. 1 shows a vapor deposition system 100 that includes a
vacuum chamber 110, an energy source 120 (e.g., an electron beam
source, an ion source, a laser beam, a magnetron source), a target 130
and a substrate 140. During operation, energy source 120 directs a
beam of energy 122 to target 130, causing material 132 to be removed
(e.g., by evaporation) from target 130 and directed to a surface 142
of substrate 140. At least a portion of the removed material 132 is
deposited on surface 142.

In general, the values of the system parameters (e.g., the temperature
of surface 142, the pressure of chamber 110, the angle of incidence of
removed material 132 on surface 142, the distance between target 130
and surface 142) can be selected as desired. The temperature of
surface 142 can be relatively low during the deposition process. For
example, during the deposition process, the ratio of the temperature
of substrate 140 to the melting point of the material forming target
130 (as determined in using Kelvin) can be about 0.5 or less (e.g.,
about 0.4 or less, about 0.35 or less, about 0.3 or less).

The pressure in chamber 110 can be relatively high. For example,
vacuum evaporation deposition, electron beam deposition or arc
evaporation, the pressure can be about 0.01 milliTorr or greater. For
gas scattering evaporation (pressure plating) or reactive arc
evaporation, the pressure in chamber 110 can be about 20 milliTorr or
greater. For sputter deposition, the pressure in chamber 110 can be
about 75 milliTorr or greater. For magnetron sputter deposition, the
pressure in chamber 110 can be about 10 milliTorr or greater. For ion
plating, the pressure in chamber 110 can be 200 milliTorr or greater.

The angle of incidence of removed material 132 on surface 142
(.theta.) can be relatively low. For example, the angle of incidence
of removed material 132 on surface 142 can be about 75.degree. or less
(e.g., about 60.degree. or less, about 45.degree. or less, about
30.degree. or less).

The distance between target 130 and surface 142 can be selected based
upon the values of the other system parameters. For example, the
distance between target 130 and surface 142 can be about 250
millimeters or less (e.g., about 150 millimeters or less, 125
millimeters or less, about 100 millimeters or less, about 90
millimeters or less, about 80 millimeters or less, about 70
millimeters or less, about 60 millimeters or less, about 50
millimeters or less, about 40 millimeters or less).

As noted above, it is believed that, the metal-containing material,
when contacted with an alcohol or water-based electrolyte, can be
released into the alcohol or water-based electrolyte (e.g., as ions,
atoms, molecules and/or clusters). It is also believed that the
ability to release the metal (e.g., as atoms, ions, molecules and/or
clusters) on a sustainable basis from a coating is generally dependent
upon a number of factors, including coating characteristics such as
composition, structure, solubility and thickness, and the nature of
the environment in which the device is used. As the level of atomic
disorder is increased, it is believed that the amount of metal species
released per unit time increases. For example, a silver metal film
deposited by magnetron sputtering at a ratio of substrate temperature
to the target melting point of less than about 0.5 and a working gas
pressure of about 0.93 Pascals (about seven milliTorr) releases
approximately 1/3 of the silver ions that a film deposited under
similar conditions, but at four Pascals (about 30 milliTorr), will
release over 10 days. Coatings formed with an intermediate structure
(e.g., lower pressure, lower angle of incidence etc.) have been
observed to have metal (e.g., silver) release values intermediate to
these values as determined by bioassays. In general, to obtain
relatively slow release of the metal, the coating should have a
relatively low degree of atomic disorder, and, to obtain relatively
fast release of the metal, the coating should have a relatively high
degree of atomic disorder.

For continuous, uniform coatings, the time for total dissolution is
generally a function of coating thickness and the nature of the
environment to which the coating is exposed. The release of metal is
believed to increase approximately linearly as the thickness of the
coating is increased. For example, it has been observed that a two
fold increase in coating thickness can result in about a two fold
increase in longevity.

In certain embodiments, it is possible to manipulate the degree of
atomic disorder, and therefore the metal release from a coating, by
forming a thin film coating with a modulated structure. For example, a
coating deposited by magnetron sputtering such that the working gas
pressure was relatively low (e.g., about two Pascals or about 15
milliTorr) for about 50% of the deposition time and relatively high
(e.g., about four Pascals or 30 milliTorr) for the remaining time, can
result in a relatively rapid initial release of metal (e.g., ions,
clusters, atoms, molecules), followed by a longer period of slow
release. This type of coating is can be particularly effective on
devices such as urinary catheters for which an initial rapid release
is advantageous to achieve quick antimicrobial concentrations followed
by a lower release rate to sustain the concentration of metal (e.g.,
ions, clusters, atoms, molecules) over a period of weeks.

It is further believed that the degree of atomic disorder of a coating
can be manipulated by introducing one or more dissimilar materials
into the coating. For example, one or more gases can be present in
chamber 110 during the deposition process. Examples of such gases
include oxygen-containing gases (e.g., oxygen, air, water), nitrogen-
containing gases (e.g., nitrogen), hydrogen-containing gases (e.g.,
water, hydrogen), boron-containing gases (e.g., boron), sulfur-
containing gases (e.g., sulfur) and halogen-containing gases (e.g.,
fluorine, chlorine, bromine, iodine). The additional gas(es) can be co-
deposited or reactively deposited with material 132. This can result
in the deposition of an oxide, nitride, carbide, boride, sulfide,
hydride and/or halide material (e.g., an oxide of a metal-containing
material, a nitride of a metal-containing material, a carbide of a
metal-containing material, a boride of a metal-containing material, a
sulfide of a metal-containing material, a hydride of a metal-
containing material, a halide of a metal-containing material). Without
wishing to be bound by theory, it is believed that atoms and/or
molecules of the additional gas(es) may become absorbed or trapped in
the material, resulting in enhanced atomic disorder. The additional
gas(es) may be continuously supplied during deposition, or may be
pulsed to (e.g., for sequential deposition). In embodiments, the
material formed can be constituted of a material with a ratio of
material 132 to additional gas(es) of about 0.2 or greater. The
presence of dissimilar atoms or molecules in the coating can enhance
the degree of atomic disorder of the coating due to the difference in
atomic radii of the dissimilar constituents in the coating.

The presence of dissimilar atoms or molecules in the coating may also
be achieved by co-depositing or sequentially depositing one or more
additional metals (e.g., one or more additional antimicrobial metals).
Such additional metals include, for example, Ta, Ti, Nb, Zn, V, Hf,
Mo, Si, Al, and other transition metals. It is believed that the
presence of dissimilar metals (one or more primary metals and one or
more additional metals) in the coating can reduce atomic diffusion and
stabilize the atomically disordered structure of the coating. A
coating containing dissimilar metals can be formed, for example, using
thin film deposition equipment with multiple targets. In some
embodiments, sequentially deposited layers of the metals are
discontinuous (e.g., islands within a the primary metal). In certain
embodiments, the weight ratio of the additional metal(s) to the
primary metal(s) is greater than about 0.2.

While FIG. 1 shows one embodiment of a deposition system, other
embodiments are possible. For example, the deposition system can be
designed such that during operation the substrate moves along rollers.
Additionally or alternatively, the deposition system may contain
multiple energy sources, multiple targets, and/or multiple substrates.
The multiple energy sources, targets and/or substrates can be, for
example, positioned in a line, can be staggered, or can be in an
array.

In certain embodiments, two layers of the material are deposited on
the substrate to achieve an optical interference effect.
Alternatively, the two layers can be formed of different materials,
with the outer (top) of the two layers being formed of an
antimicrobial, atomically disordered, nanocrystalline silver-
containing material, and the inner of the two layers having
appropriate reflective properties so that the two layers can provide
an interference effect (e.g., to monitor the thickness of the outer
(top) of the two layers).

The substrate can be selected as desired. The substrate may be formed
of one layer or multiple layers, which may be formed of the same or
different materials.

In certain embodiments, the substrate can include one or more layers
containing a bioabsorbable material. Bioabsorbable materials are
disclosed, for example, in U.S. Pat. No. 5,423,859. In general,
bioabsorbable materials can include natural bioabsorbable polymers,
biosynethetic bioabsorbable polymers and synthetic bioabsorbable
polymers. Examples of synthetic bioabsorbable polymers include
polyesters and polylactones (e.g., polymers of polyglycolic acid,
polymers of glycolide, polymers of lactic acid, polymers of lactide,
polymers of dioxanone, polymers of trimethylene carbonate,
polyanhydrides, polyesteramides, polyortheoesters, polyphosphazenes,
and copolymers of the foregoing). Examples of natural bioabsorbable
polymers include proteins (e.g., albumin, fibrin, collagen, elastin),
polysaccharides (e.g., chitosan, alginates, hyaluronic acid). Examples
of biosynthetic polymers include polyesters (e.g., 3-hydroxybutyrate
polymers).

In some embodiments, the substrate includes multiple layers (e.g., two
layers, three layers, four layers, five layers, six layers, seven
layers, eight layers, nine layers, 10 layers). The layers can be
laminated together (e.g., by thermal fusing, stitching and/or
ultrasonic welding).

One or more layers (e.g., an outer layer) of a multi-layer substrate
can be formed of a perforated (and optionally non-adherent) material
(e.g., a woven material or a non-woven material) that can allow fluid
to penetrate or diffuse therethrough. Such materials include, for
example, cotton, gauze, polymeric nets (e.g., polyethylene nets, nylon
nets, polypropylene nets, polyester nets, polyurethane nets,
polybutadiene nets), polymeric meshes (e.g., polyethylene meshes,
nylon meshes, polypropylene meshes, polyester meshes, polyurethane
meshes, polybutadiene meshes) and foams (e.g., an open cell
polyurethane foam). Examples of commercially available materials
include DELNET.TM. P530 non-woven polyethylene veil (Applied Extrusion
Technologies, Inc., Middletown, Del.), Exu-Dry CONFORMANT2.TM. non-
woven polyethylene veil (Frass Survival Systems, Inc., NY, N.Y.),
CARELLE.TM. material (Carolina Formed Fabrics Corp.), NYLON90.TM.
material (Carolina Formed Fabrics Cop.), N-TERFACE.TM. material
(Winfield Laboratories, Inc., Richardson, Tex.), HYPOL.TM. hydrophilic
polyurethane foam (W. R. Grace & Co., NY, N.Y.).

One or more layers (e.g., an inner layer) of a multi-layer substrate
can be formed of an absorbent material (e.g., a woven material or a
non-woven material) formed of, for example, rayon, polyester, a rayon/
polyester blend, polyester/cotton, cotton and/or cellulosic fibers.
Examples include creped cellulose wadding, air felt, air laid pulp
fibers and gauze. An example of a commercially available material is
SONATRA.TM. 8411 70/30 rayon/polyester blend (Dupont Canada,
Mississauga, Ontario).

One or more layers (e.g., an outer layer) of a multi-layer substrate
can be formed of an occlusive or semi-occlusive material, such as an
adhesive tape or polyurethane film (e.g., to secure the device to the
skin and/or to retain moisture).

In some embodiments, the layers in a multi-layer substrate are
laminated together (e.g., at intermittent spaced locations) by
ultrasonic welds. Typically, heat (e.g., generated ultrasonically) and
pressure are applied to either side of the substrate at localized
spots through an ultrasonic horn so as to cause flowing of at least
one of the plastic materials in the first and second layers and the
subsequent bonding together of the layers on cooling. The welds can be
formed as localized spots (e.g., circular spots). The spots can have a
diameter of about 0.5 centimeter or less.

The shape of the substrate can generally be varied as desired. For
example, the substrate can be in the shape of a film, a fiber or a
powder.

The substrate/coating article can be used in a variety of articles.
For example, the article can be in the shape of a medical device.
Exemplary medical devices include wound closure devices (e.g.,
sutures, staples, adhesives), tissue repair devices (e.g., meshes,
such as meshes for hernia repair), prosthetic devices (e.g., internal
bone fixation devices, physical barriers for guided bone
regeneration), tissue engineering devices (e.g., for use with a blood
vessel, skin, a bone, cartilage, a liver), controlled drug delivery
systems (e.g., microcapsules, ion-exchange resins) and wound coverings
and/or fillers (e.g., alginate dressings, chitosan powders). In some
embodiments, the article is a transcutaneous medical device (e.g., a
catheter, a pin, an implant), which can include the substrate/coating
supported on, for example, a solid material (e.g., a metal, an alloy,
latex, nylon, silicone, polyester and/or polyurethane). In some
embodiments, the article is in the form of a patch (e.g., a patch
having an adhesive layer for adhering to the skin, such as a
transdermal patch).

Subsequent to deposition, the material can optionally be annealed. In
general, the anneal is conducted under conditions to increase the
stability (e.g., shelf life) of the material while maintaining the
desired therapeutic activity of the material. In certain embodiments,
the material can be annealed at a temperature of about 200.degree. C.
or less (e.g., about room temperature).

The substrate/coating is typically sterilized prior to use (e.g.,
without applying sufficient thermal energy to anneal out the atomic
disorder). The energy used for sterilization can be, for example,
gamma radiation or electron beam radiation. In some embodiments,
ethylene oxide sterilization techniques are used to sterilize the
substrate/coating.

Free Standing Powders

A free standing powder can be prepared by, for example, cold working
or compressing to impart atomic disorder to the powder. In certain
embodiments, a free standing powder is prepared by forming a coating
of the material as described above, and then removing the material
from the surface of the substrate. For example, the material can be
scraped from the surface of the substrate by one or more scrapers. In
embodiments in which the substrate moves during deposition of the
material, the scrapers can remove the material as the substrate moves.
The scrapers can be, for example, suspended above the substrate. Such
scrapers can be, for example, weighted and/or spring loaded to apply
pressure sufficient to remove the material as the substrate moves. In
some embodiments (e.g., when a continuous belt is used), the scrapers
can be located above the end rollers to remove the material with a
reverse dragging action as the substrate rounds the end roller.

A free standing powder can be used to treat a condition in various
ways. As an example, the powder can sprinkled onto the subject's skin.
As another example, the powder can be inhaled using an inhaler, such
as a dry powder inhaler.

In certain embodiments (e.g., when the free standing powder is
inhaled), the average particle size of the free standing powder is
selected to reduce the likelihood of adverse reaction(s) of the
particles in the tissue (e.g., tissue contacted by the free standing
powder during inhalation). In embodiments, a free standing powder can
have an average particle size of less than about two microns (e.g.,
less than about one micron, less than about 0.5 micron).

Powder Impregnated Materials

The metal-containing material can be in the form of a powder
impregnated material. Such powder impregnated materials can, for
example, be in the form of a hydrocolloid having the free standing
powder blended therein. A powder impregnated material can be, for
example, in the form of a dressing, such as a hydrocolloid dressing.

Solutions

The compound can be in the form of a solution (e.g., a solvent-based
solution). The solution can be formed, for example, by dissolving a
free standing powder of the material in a solvent for the powder. As
an example, a container (e.g., a tea bag-type container) with the free
standing powder within it can be immersed in the water or solvent. As
another example, a substrate (e.g., in the form of a strip or a
bandage) carrying the material can be immersed in the solvent. In
certain embodiments, it can be preferable to form a solution by
dissolving a free standing powder of the compound in a solvent because
this can be a relatively approach to forming a solution.

In certain embodiments, the solution containing the compound is
contacted with the subject relatively soon after formation of the
solution. For example, the solution containing the compound can be
contacted with the subject within about one minute or less (e.g.,
within about 30 seconds or less, within about 10 seconds or less) of
forming the solution containing the compound. In some embodiments, a
longer period of time lapses before the solution containing the
compound is contacted with the subject. For example a period of time
of at least about 1.5 minutes (e.g., at least about five minutes, at
least about 10 minutes, at least about 30 minutes, at least about one
hour, at least about 10 hours, at least about a day, at least about a
week) lapses between the time the solution containing the compound is
formed and the solution containing the compound is contacted with the
subject.

In some embodiments, lowering the pH of the solution (e.g., to less
than about 6.5, such as from about 3.5 to about 6.5) can allow for a
higher concentration of the dissolved material and/or a faster rate of
dissolution. The pH of the solution can be lowered, for example, by
adding acid to the solution (e.g., by adding CO.sub.2 to the solution
to form carbonic acid).

A solution containing the compound can be contacted with the subject
with or without the use of a device. As an example, a solution
containing the compound can be contacted with the skin, mouth, ears or
eyes as a rinse, a bath, a wash, a gargle, and/or drops. As another
example, the solution can be injected using a small needle injector
and/or a needleless injector. As an additional example, a solution
containing the compound can be formed into an aerosol (e.g., an
aerosol prepared by a mechanical mister, such as a spray bottle or a
nebulizer), and the aerosol can be contacted with the subject using an
appropriate device (e.g., a hand held inhaler, a mechanical mister, a
spray bottle, a nebulizer, an oxygen tent). As a further example, a
solution containing the compound can be contacted with the second
location via a catheter.

In embodiments in which onychomycosis is being treated, the method can
include first hydrating the nail with urea (1-40%) or lactic acid
(10-15%), followed by treatment with the metal-containing material,
which may contain an appropriate solvent (e.g., DMSO) for penetration
through the nail. Alternatively or additionally, onychomycosis can be
treated by injecting (e.g., via a needleless injector and/or a needle)
the metal-containing material to the affected area.

Typically, the solvent is a relatively hydrophilic solvent. Examples
of solvents include water, DMSO and alcohols. In certain embodiments,
a water-based solution is a buffered solution. In some embodiments, a
water-based solution contains carbonated water. In embodiments, more
than one solvent can be used.

In some embodiments, the solution can contain about 0.001 weight
percent or more (e.g., about 0.01 weight percent or more, about 0.02
weight percent or more, about 0.05 weight percent or more, about 0.1
weight percent or more, about 0.2 weight percent or more, about 0.5
weight percent or more, about one weight percent or more) of the
compound and/or about 10 weight percent or less (e.g., about five
weight percent or less, about four weight percent or less, about three
weight percent or less, about two weight percent or less, about one
weight percent or less) of the compound.

Pharmaceutical Carrier Compositions

The metal-containing material can disposed (e.g., suspended) within a
pharmaceutically acceptable carrier. The formulation can be, for
example, a semi-solid, a water-based hydrocolloid, an oil-in-water
emulsion, a water-in-oil emulsion, a non-dried gel, and/or a dried
gel. Typically, when disposed in a pharmaceutically acceptable
carrier, the metal-containing material is applied to the skin.

Examples of pharmaceutically acceptable carriers include creams,
ointments, gels, lotions, pastes, foams and liposomes.

The formulation can contain about 0.01 weight percent or more (e.g.,
about 0.1 weight percent or more, about 0.5 weight percent or more,
about 0.75 weight percent or more, about one weight percent or more,
about two weight percent or more, about five weight percent or more,
about 10 weight percent or more) of the metal-containing material and/
or about 50 weight percent or less (e.g., about 40 weight percent or
less, about 30 weight percent or less, about 20 weight percent or
less, about 20 weight percent or less, about 15 weight percent or
less, about 10 weight percent or less, about five weight percent or
less) of the metal-containing material.

In certain embodiments, the metal-containing material can be
effectively used in the oral cavity when in the form of an article
(e.g., a tape, a pill, a capsule, a tablet or lozenge) that is placed
within the oral cavity (e.g., so that the subject can suck on the
tape, pill, capsule, tablet or lozenge). In some embodiments, the
article can be a sustained release article (e.g., a sustained release
capsule) In certain embodiments, the article can be an enteric article
(e.g., an enteric coated tablet).

Formulations can optionally include one or more components which can
be biologically active or biologically inactive. Examples of such
optional components include base components (e.g., water and/or an
oil, such as liquid paraffin, vegetable oil, peanut oil, castor oil,
cocoa butter), thickening agents (aluminum stearate, hydrogen
lanolin), gelling agents, stabilizing agents, emulsifying agents,
dispersing agents, suspending agents, thickening agents, coloring
agents, perfumes, excipients (starch, tragacanth, cellulose
derivatives, polyethylene glycols, silicones, bentonites, silicic
acid, talc), foaming agents (e.g., surfactants), surface active
agents, preservatives (e.g., methyl paraben, propyl paraben) and
cytoconductive agents (e.g., betaglucan). In certain embodiments, a
pharmaceutical carrier composition can include a constituent (e.g.,
DMSO) to assist in the penetration of skin.

While the foregoing has described embodiments in which a single
condition is treated, in some embodiments multiple conditions can be
treated. The multiple conditions can be the same type of condition
(e.g., multiple skin or integument conditions) or different types of
conditions. For example, a dressing formed of one or more substrates
coated with an appropriate metal-containing material (e.g.,
antimicrobial, atomically disordered, silver-containing material) can
be applied to an area of the skin having multiple skin or integument
conditions (e.g., a burn and psoriasis) so that the metal-containing
material treats the multiple skin or integument conditions.

Moreover, while the foregoing has described embodiments that involve
one method of contacting a subject with the metal-containing material,
in other embodiments, more than one method of contacting a subject
with the metal-containing material can be used. For example, the
methods can include one or more of ingestion (e.g., oral ingestion),
injection (e.g., using a needle, using a needleless injector), topical
administration, inhalation (e.g., inhalation of a dry powder,
inhalation of an aerosol) and/or application of a dressing.

Furthermore, while the foregoing has described embodiments in which
one form of the metal-containing material is used, in other
embodiments, more than one form of the metal-containing material can
be used. For example, the methods can include using the metal-
containing material in the form of a coating (e.g., a dressing), a
free standing powder, a solution and/or a pharmaceutical carrier
composition.

Moreover, the metal-containing material can be used in various
industrial applications. For example, the metal-containing material
can be used to reduce and/or prevent microbial growth on industrial
surfaces (e.g., industrial surfaces where microbial growth may occur,
such as warm and/or moist surfaces). Examples of industrial surfaces
include heating pipes and furnace filters. In certain embodiments, the
metal-containing material can be disposed (e.g., coated or sprayed) on
the surface of interest to reduce and/or prevent microbial growth.
This can be advantageous in preventing the spread of microbes via, for
example, heating and/or air circulation systems within buildings.

The following examples are illustrative and not intended as limiting.

EXAMPLES

Treatment of Hyperproliferative Skin Conditions

Example 1

Preparation of Nanocrystalline Silver Coatings on Dressings

This example shows the preparation of a bilayer nanocrystalline silver
coating on a dressing material. A high density polyethylene dressing,
DELNET.TM. or CONFORMANT 2.TM. was coated with a silver base layer and
a silver/oxide top layer to generate a coloured anti-microbial coating
having indicator value. The coating layers were formed by magnetron
sputtering under the conditions set out in the following table.

TABLE-US-00001 Sputtering Conditions: Base Layer Top Layer Target
99.99% Ag 99.99% Ag Target Size 20.3 cm diameter 20.3 cm diameter
Working Gas 96/4 wt % Ar/O.sub.2 96/4 wt % Ar/O.sub.2 Working Gas
Pressure 5.33 Pa (40 mT) 5.33 Pa (40 mT) Power 0.3 kW 0.15 kW
Substrate Temperature 20.degree. C. 20.degree. C. Base Pressure
3.0 .times. 10.sup.-6 Torr 3.0 .times. 10.sup.-6 Torr Anode/Cathode
Distance 100 mm 100 mm Sputtering Time 7.5-9 min 1.5 min Voltage
369-373 V 346 V

The resulting coating was blue in appearance. A fingertip touch was
sufficient to cause a colour change to yellow. The base layer was
about 900 nm thick, while the top layer was 100 nm thick.

To establish that silver species were released from the coated
dressings, a zone of inhibition test was conducted. Mueller Hinton
agar was dispensed into Petri dishes. The agar plates were allowed to
surface dry prior to being inoculated with a lawn of Staphylococcus
aureus ATCC#25923. The inoculant was prepared from Bactrol Discs
(Difco, M.), which were reconstituted as per the manufacturer's
directions. Immediately after inoculation, the coated materials to be
tested were placed on the surface of the agar. The dishes were
incubated for 24 hr. at 37.degree. C. After this incubation period,
the zone of inhibition was calculated (corrected zone of
inhibition=zone of inhibition-diameter of the test material in contact
with the agar). The results showed a corrected ZOI of about 10 mm,
demonstrating good release of silver species.

The coating was analyzed by nitric acid digestion and atomic
absorption analysis to contain 0.24+/-0.04 mg silver per mg high
density polyethylene. The coating was a binary alloy of silver (>97%)
and oxygen with negligible contaminants, based on secondary ion mass
spectroscopy. The coating, as viewed by SEM, was highly porous and
consisted of equiaxed nanocrystals organized into coarse columnar
structures with an average grain size of 10 nm. Silver release studies
in water demonstrated that silver was released continuously from the
coating until an equilibrium concentration of about 66 mg/L was
reached (determined by atomic absorption), a level that is 50 to 100
times higher than is expected from bulk silver metal
(solubility .ltoreq.1 mg/L).

By varying the coating conditions for the top layer to lengthen the
sputtering time to 2 min, 15 sec., a yellow coating was produced. The
top layer had a thickness of about 140 nm and went through a colour
change to purple with a fingertip touch. Similarly, a purple coating
was produced by shortening the sputtering time to 1 min, to achieve a
top layer thickness of about 65 nm. A fingertip touch caused a colour
change to yellow.

To form a three layer dressing, two layers of this coated dressing
material were placed above and below an absorbent core material formed
from needle punched rayon/polyester (SONTARA.TM. 8411). With the
silver coating on both the first and third layers, the dressing may be
used with either the blue coating side or the silver side in the skin
facing position. For indicator value, it might be preferable to have
the blue coating visible. The three layers were laminated together by
ultasonic welding to produce welds between all three layers spaced at
about 2.5 cm intervals across the dressing. This allowed the dressing
to be cut down to about 2.5 cm size portions for smaller dressing
needs while still providing at least one weld in the dressing portion.

The coated dressings were sterilized using gamma radiation and a
sterilization dose of 25 kGy. The finished dressing was packaged
individually in sealed polyester peelable pouches, and has shown a
shelf life greater than 1 year in this form. The coated dressings can
be cut in ready to use sizes, such as 5.1.times.10.2 cm strips, and
slits formed therein before packaging. Alternatively, the dressings
may be packaged with instructions for the clinician to cut the
dressing to size and form the desired length of the slit for the
medical device.

Additional silver coated dressings were prepared in a full scale roll
coater under conditions to provide coatings having the same properties
set out above, as follows: the dressing material included a first
layer of silver coated DELNET, as set out above, laminated to STRATEX,
AET, 8.0NP.sub.2-A/QW, which is a layer of 100% rayon on a
polyurethane film. Silver Foam Dressing--three layers of silver coated
high density polyethylene prepared as above, alternating with two
layers of polyurethane foam, L-00562-6 Medical Foam, available from
Rynel Ltd., Bootbay, Me., USA.

Example 2

Preparation of Nanocrystalline Silver Powders

Nanocrystalline silver powder was prepared by preparing silver
coatings on silicon wafers, under the conditions set forth in the
table above, and then scraping the coating off using a glass blade.

Nanocrystalline silver powder was also prepared by sputtering silver
coatings on silicon wafers using Westaim Biomedical NGRC unit, and
then scraping the coating off. The sputtering conditions were as
follows:

TABLE-US-00002 Target: 99.99% Ag Target Size: 15.24 cm .times.
1216.125 cm Working Gas: 75:25 wt % Ar/O.sub.2 Working Gas Pressure:
40 mTorr Total Current: 40 A Base Pressure: 5.0 .times. 10.sup.-5 Torr
Sandvik Belt Speed: 340 mm/min Voltage: 370 V

The powder has a particle size ranging from 2 .mu.m to 100 .mu.m, with
crystallite size of 8 to 10 nm, and demonstrated a positive rest
potential.

Example 3

Treatment of Psoriasis

This patient was a 58 year old female with psoriatic plaques covering
up to sixty percent of her body. For this patient, psoriatic plaques
first occurred ten years ago and have been treated with the following:

1. Adrenal corticosteroids. Injections gave relief from pruritus and
general discomfort. Treatments led to a rebound effect; i.e. psoriasis
would flare up after treatments wore off. Corticosteroids were
discontinued.

2. UV Light and Methotrexate treatments. UV light treatments were
given in conjunction with methotrexate. The UV light treatments caused
burns and new lesions. The methotrexate caused severe nausea. Both
treatments were discontinued.

3. Ice Cap Spray. This treatment contained a potent corticosteroid,
and gave some relief but it was taken off the market and is no longer
available.

4. Soriatone (acetretin 10 mg). This systemic retinoid treatment was
associated with joint aches and was discontinued.

5. Diet. The patient was attempting to control the disease through
diet.

Nanocrystalline silver was tested as follows. Nanocrystalline silver
was deposited on sheets of high-density polyethylene (HDPE) using a
vapour deposition process as set forth in

Example 1. Two sheets of this coated HDPE were laminated together
around a core of non-woven rayon polyester, as set forth in Example 1.
A 50 mm.times.50 mm (2''.times.2'') piece of this composite material
was saturated with water and placed centrally on a one and a half year
old 150 mm.times.100 mm (6''.times.4'') psoriatic plaque on the
patient's flank. The nanocrystalline silver coated material was
covered with a piece of low moisture vapour transmission thin polymer
film. The polymer sheet extended 50 mm (2'') beyond the
nanocrystalline silver coated HDPE to provide control data regarding
occlusion of the psoriatic plaque.

The dressing was removed after three days. There was no discernible
change in the plaque at this time. However two days later the area
that was covered with the nanocrystalline silver had the appearance of
normal skin while the rest of the plaque was still rough and
unchanged, including the untreated but occluded area.

The nanocrystalline silver therapy caused the treated psoriatic plaque
to resolve.

Example 4

Treatment of Psoriasis

The subject was a 58 year old female with psoriatic plaques over up to
sixty percent of her body. Psoriatic plaques had first occurred 10
years ago and had been treated with the following:

1. Adrenal corticosteroids. Injections gave relief from pruritus and
general discomfort. Treatments led to a rebound effect i.e. psoriasis
would flare up after treatments wore off. Corticosteroids were
discontinued.

2. UV Light and Methotrexate treatments. UV light treatments were
given in conjunction with methotrexate. The UV light treatments caused
bums and new lesions. The methotrexate caused severe nausea. Both
treatments were discontinued.

3. Ice Cap Spray. This treatment contained a potent corticosteroid,
and gave some relief but it was taken off the market and is no longer
available.

4. Soriatone (acetretin 10 mg). This systemic retinoid treatment was
associated with joint aches and was discontinued.

5. Diet. The patient was attempting to control the disease through
diet.

Nanocrystalline silver was tested as follows. Nanocrystalline silver
was deposited on sheets of high-density polyethylene (HDPE) using a
vapour deposition process as set forth in Example 1 (top layer). Two
sheets of this coated HDPE were laminated together around a core of
non-woven rayon polyester, as set forth in Example 1. A 50 mm.times.50
mm (2''.times.2'') piece of this composite material was saturated with
water and placed centrally on a 125 mm.times.100 mm (5''.times.4'')
psoriatic plaque on the patient's upper left thigh. The
nanocrystalline silver coated material was covered with a piece of low
moisture vapour transmission thin polymer film. The polymer sheet
extended 50 mm (2'') beyond the nanocrystalline silver coated HDPE to
provide control data regarding occlusion of the psoriatic plaque.

The dressing was removed and the plaque examined after two days. The
area that was covered with the nanocrystalline silver was free of
scaling and only slightly erythematous while the rest of the plaque
was still erythenatous and scaly, including the untreated but occluded
area. The plaque was redressed with a similar 50 mm.times.50 mm
(2''.times.2'') piece of nanocrystalline silver coated dressing, which
was left in place for a further period of 2 days. The area that was
covered with the nanocrystalline silver remained free of scale and
only slightly erythenatous, while the rest of the plaque was still
erythenatous and scaly, including the area under the occlusive film.

The nanocrystalline silver therapy caused the treated psoriatic plaque
to resolve.

Example 5

Preparation of Nanocrystalline Gels

A commercial carboxymethyl cellulose/pectin (Duoderm Convatec.TM.) was
combined with nanocrystalline silver powder prepared as in Example 2
to produce a gel with 0.1% w/v. silver. Carboxymethyl cellulose (CMC)
fibers were coated by magnetron sputtering, under conditions similar
to those set out in Example 1 for the top layer to produce a defective
nanocrystalline silver coating. The CMC was then gelled in water by
adding 2.9 g to 100 mL volume. An alginate fibrous substrate was
directly coated with a defective nanocrystalline silver coating by
magnetron sputtering under coating conditions similar to those set
forth in Example 1 for the top layer. The alginate (5.7 g) was added
to 100 mL volume of water to create a gel. A commercial gel containing
CMC and alginate (Purilon gel Coloplast.TM.) was mixed with an atomic
disordered nanocrystalline silver powder prepared as in Example 2 to
give a gel product with 0.1% w/v silver. A commercially available gel
(Lubriderm.TM.--glyceryl polymethacrylate) was blended with atomic
disordered nanocrystalline silver powder prepared as in Example 2, to
prepare a gel with a silver content of 0.1% w/v. A further gel was
formulated with, on w/v basis, 0.1% methyl paraben, 0.02% propyl
paraben, 0.5% polyvinyl alcohol (Airvol.TM. PVA 540), 2% CMC, 0.1%
nanocrystalline silver powder prepared as in Example 2, and was
brought up to 1000 g with water.

Treatment of Inflammatory Skin Conditions

Example 1

Preparation of Nanocrystalline Silver Coatings on Dressings

This example shows the preparation of a bilayer nanocrystalline silver
coating on a dressing material. A high density polyethylene dressing,
DELNET.TM. or CONFORMANT 2.TM. was coated with a silver base layer and
a silver/oxide top layer to generate a coloured antimicrobial coating
having indicator value as described in Example 1 of the Treatment of
Hyperproliferative Skin conditions examples. The coating layers were
formed by magnetron sputtering under the conditions set out in the
following table.

Example 2

Preparation of Nanocrystalline Silver Coating on HDPE Mesh

The silver coated mesh was produced, as set forth in Example 1, by
sputtering silver onto Delnet, a HDPE mesh (Applied Extrusion
Technologies, Inc., Middletown, Del., USA) using Westaim Biomedical
TMRC unit under the following conditions:

TABLE-US-00003 Target: 99.99% Ag Target Size: 15.24 cm .times. 152.4
cm Working Gas: 99.375:0.625 wt % Ar/O.sub.2 Working Gas Pressure:
5.33 Pascals (40 mTorr) Total Current: 22 A Base Pressure: 5.0 .times.
10.sup.-5 Torr Sandvik Belt Speed: 577 mm/min Voltage: 367 V

The coating was tested and found to have a weight ratio of reaction
product to silver of between 0.05 and 0.1. The dressing was non-
staining to human skin.

Example 3

Preparation of Atomic Disordered Nanocrystalline Silver Powders

Nanocrystalline silver coatings were prepared by sputtering silver in
an oxygen-containing atmosphere directly onto an endless stainless
steel belt of a magnetron sputtering roll coater, or onto silicon
wafers on the belt. The belt did not need to be cooled. The coatings
were scraped off with the belt with suspended metal scrapers as the
belt rounded the end rollers. For the coated silicon wafers, the
coatings were scraped off with a knife edge. The sputtering conditions
were as follows:

TABLE-US-00004 Target: 99.99% Ag Target Size: 15.24 cm .times.
1216.125 cm Working Gas: 75:25 wt % Ar/O.sub.2 Working Gas Pressure:
5.33 Pascals (40 milliTorr) Total Current: 40 A Base Pressure:
5.0 .times. 10.sup.-5 Torr (range: 1 .times. 10.sup.-4-9 .times.
10.sup.-7 Torr or 1 .times. 10.sup.-2-1.2 .times. 10.sup.-4 Pa)
Sandvik Belt Speed: 340 mm/min Voltage: 370 V Note - pressure
conversions to Pa herein may not be accurate, most accurate numbers
are in torr, mTorr units.

The powder had a particle size ranging from 2 .mu.m to 100 .mu.m, with
grain or crystallite size of 8 to 10 nm (i.e., nanocrystalline), and
demonstrated a positive rest potential.

Similar atomic disordered nanocrystalline silver powders were formed
as set forth hereinabove by magnetron sputtering onto cooled steel
collectors, under conditions taught in the prior Burrell et al.
patents to produce atomic disorder.

Example 4

In vitro Activity of Silver Solution against Propionibacterium acne

An in vitro test was conducted to determine if silver solutions
according to the present invention effectively control
Propionibacterium acne. The silver solution was obtained by static
elution of Acticoat.TM. Burn Wound Dressing (lot #: 00403A-05, Westaim
Biomedical Corp., Fort Saskatchewan, Canada) with nanopure water in a
ratio of one square inch of dressing in five milliliters of water for
24 hours at room temperature. The silver concentration of the silver
solution was determined by an atomic absorption method. The silver
elute was diluted with nanopure water to 20 .mu.g/ml. The
Propionibacterium acne (ATCC No. 0919) was provided by Biofilm
Research Group, University of Calgary.

The inoculum was prepared by inoculating freshly autoclaved and cooled
tubes of Tryptic soy broth (TSB) with P. acne and incubating them for
2 days at 37.degree. C. in an anaerobic jar. At this time, the optical
density of the suspensions was .about.0.3 at a wavelength of 625 nm.

The bacterial suspension (100 .mu.L) was mixed with 100 .mu.L of the
silver solution being tested. The final concentration of silver in
these mixtures was 10 .mu.g/ml. The mixtures were incubated in an
anaerobic jar at 37.degree. C. for two hours. The silver was
neutralized by addition of 0.4% STS (0.85% NaCl, 0.4% Sodium
thioglycolate, 1% Tween.TM. 20) and the solution was serially 10-fold
diluted with phosphate-buffered saline. 20 .mu.L aliquots of the
original solution and subsequent dilutions were plated onto TSA drop
plates. The drops were allowed to dry and the plates were incubated in
an anaerobic jar at 37.degree. C. for 72 hours at which time the
colonies were counted. The control consisted of 100 .mu.L of bacterial
suspension mixed with 100 .mu.L of nanopure water and treated as
above.

The results showed that the silver solution according to the present
invention, at a final concentration of 10 .mu.g/ml, gave 4.3 logarithm
reduction in viable P. acne counts in two hours.

Example 5

Treatment of Acne

A sixteen year old female was diagnosed with acne vulgaris. She had
numerous red papules and pustules on her forehead. Various skin
cleansing regimes and antibiotic (erythromycin and clindomycin)
treatments had been tried and had failed to control the acne. Prior to
bedtime, the papules and pustules on one side of her forehead were
moistened and covered with a nanocrystalline silver coated high
density polyethylene mesh, prepared as in Example 1 (single layer,
blue coating). The mesh was then occluded with a thin film dressing
which remained in place for 10 hours. Upon removal, the papules and
pustules were no longer red and were only slightly raised. Some brown
staining of the skin was observed.

Example 6

Treatment of Acne

A sixteen year old male was diagnosed with acne vulgaris. He had
numerous raised, red papules and pustules on his forehead. Various
skin cleansing regimes and antibiotic treatments had been tried and
had failed to control the acne. The patient was placed on isotretinoin
treatment which controlled his acne well. He did develop a single
large pustule on his forehead which was embarrassing for him. Prior to
bedtime, the pustule was moistened and covered with a nanocrystalline
silver coated high density polyethylene mesh prepared as in Example 2.
The mesh was then occluded with a thin film dressing which remained in
place for 10 hours. Upon removal the pustule was no longer red and was
only slightly raised. A second treatment resulted in the disappearance
of the pustule.

Example 7

Treatment of Acne

A sixteen year old female was diagnosed with acne vulgaris. She had
numerous red papules and pustules on her forehead. Various skin
cleansing regimes and antibiotic (erythromycin and clindomycin)
treatments had been tried and had failed to control the acne. Prior to
bedtime, the papules and pustules on one side of her forehead were
moistened and covered with a nanocrystalline silver coated high
density polyethylene mesh, prepared as in Example 2. The mesh was then
occluded with a thin film dressing which remained in place for 10
hours. Upon removal the papules and pustules were no longer red and
were only slightly raised. A second treatment resulted in the
disappearance of the papules and virtual elimination of the pustules.
The silver coated mesh, when prepared as set forth in Example 2, did
not result in any staining of the skin.

Example 8

Treatment of Adult Acne with Silver-Impregnated Hydrocolloid Dressing

A 49 year old white male experienced occasional acne vulgaris. He had
painful, raised, red papules and pustules on his shoulders. The
patient was treated with a thin hydrocolloid dressing (Craig Medical
Products Ltd., Clay Gate House 46 Albert Rd. North Reigate, Surrey,
United Kingdom) which was impregnated with 1% nanocrystalline silver
powder formed with atomic disorder as in Example 3. Following
cleansing, the pustule was covered with a small disc of the dressing,
which remained in place for 24 hours. Upon removal, the pustule was no
longer painful, red, or raised.

Example 9

Treatment of Eczema

A twenty-nine year old white female presented with acrodermatitis. The
erythematous area was located on the dorsal surface of the first web
space of the left hand. It was bounded by the metacarpal bones of the
thumb and index finger. The patient also complained of pruritus
associated with the dermatitis. A gel consisting of 0.1%
nanocrystalline silver powder (formed with atomic disorder as in
Example 3) and 2% carboxymethylcellulose was applied to the inflamed
area before bedtime. There was an immediate antipruritic effect that
provided the patient with relief in the short term. The next morning
all evidence of acrodermatitis (i.e. redness disappeared) was gone.
The condition had not returned after two weeks.

Example 10

Allergic Contact Dermatitis

Skin allergic contact hypersensitivity is caused by excessive
infiltration of eosinophils. An animal model may be used for in vivo
evaluation of eosinophil infiltration in the contact sensitivity
reaction and to determine whether it is associated with allergic skin
conditions such as contact dermatitis. On a gross histology level,
this can be measured by the degree of erythema and edema at the
dermatitis site. Current drugs used for treatment of this and other
related eczema conditions include high potency steroids
(Ultravate.TM.), medium potency steroids (Elocon.TM.) and non
steroidal anti-inflammatory compounds (Protopic.TM. or tacrolimus).
These compounds do not always work and may have undesirable side
effects. Several commercially available anti-inflammatory products
were compared to a nanocrystalline silver powder for the treatment of
allergic contact dermatitis as follows.

Four healthy domestic pigs (approximate weight 20 kg) were used in the
study. All pigs had normal skin prior to induction of eczema with 10%
2,4-dinitrochlorobenzene (DNCB) in acetone. The animals were housed in
appropriate animal facilities with 12 hour light-dark cycles. The pigs
were fed antibiotic-free feed and water ad libitum. The pigs were
housed and cared for in accordance with Canadian Council of Animal
Care guidelines. On day 0, the hair on both left and right back and
side were clipped. The DNCB solution was painted over this area. This
was repeated on day 7 and 11. On day 11, the solution was painted
approximately 4 hours before treatment was initiated.

Treatment groups are shown in the following table. Protopic.TM.
(tacrolimus), Elocon.TM. and Ultravate.TM. were purchased as creams
from the local pharmacy. The nanocrystalline silver powder (1 g/L) was
mixed into a 2% sodium carboxymethyl cellulose (CMC) and water
solution at 30.degree. C. using a magnetic stirrer at a high speed
(Vista Scientific). Petrolatum, commercially known as Vaseline.TM.,
was used as a control for Elocon.TM. and Ultravate.TM..

TABLE-US-00005 Day of Pig # Treatment (Left Side) Control (Right Side)
Treatment 1 Protopic .TM. (tacrolimus) Protopic .TM. Control Day 0 2
Medium Potency Steroid Petrolatum Day 0 (Elocon .TM.) 3 2% CMC + 1% 2%
CMC Day 0 nanocrystalline silver (Vista Scientific) 4 High Potency
Steroid Petrolatum Day 0 (Ultravate .TM.)

Pigs were placed under general anesthetic with ketamine (Ketalean.TM.,
MTC Pharmaceuticals, Cambridge, ON; 4-500 mg) and halothane (MTC
Pharmaceuticals). The skin was wiped with a moist gauze and allowed to
dry. Bandages (n=8) containing each treatment were applied to the left
side of the thoracolumbar area of the pig, while control bandages
(n=8) were applied to the right side of the thoracolumbar area of the
pig. Following placement of bandages, they were covered with
Tegaderm.TM. (3M Corp., Minneapolis, Minn.) which was secured with an
Elastoplast.TM. (Smith and Nephew, Lachine, QC) wrap. Bandages with
active agents were changed daily. The skin associated with each
bandage site was scored for severity of erythema (0=normal, 1=slight,
2=moderate, 3=severe, 4=very severe) and swelling (0=normal, 1=slight,
2=moderate, 3=severe, 4=very severe). This was performed on days 0, 1,
2 and 3.

All pigs remained healthy during the study. Results are shown in the
following tables, and indicated in FIGS. 3 and 4. FIGS. 3 and 4 show
the efficacy of the nanocrystalline silver powder compared to
Protopic.TM., Elocon.TM. and Ultravate.TM. in the treatment of contact
dermatitis in the pig model.

TABLE-US-00006 Day 0 Day 1 Day 2 Day 3 Treatment (Erythema)
Nanocrystalline silver 3 2 1 0 Protopic .TM. 3 3 1.9 0.4 Elocon .TM. 3
2.4 2.6 2.6 Ultravate .TM. 3 3 3 3 Treatment (Edema) Nanocrystalline
silver 2 1 0 0 Protopic .TM. 2 2 2 0 Elocon .TM. 2 2 2 0
Ultravate .TM. 2 2 1 0

The pigs treated with the high (Ultravate.TM.) and medium (Elocon.TM.)
strength steroids showed little to no improvement in the degree of
erythema associated with contact dermatitis. They did, however,
improve in terms of edema in that at Day 3, no swelling was apparent.
Protopic.TM. showed a marked improvement when compared to the steroids
in both the degree of erythema and edema. The largest improvement
occurred with the nanocrystalline silver powder suspended in a 2%
carboxymethyl cellulose gel. Both erythema and edema scores were lower
after a single treatment and were normal after Day 2 (edema) and Day 3
(erythema) of treatment. Clearly the nanocrystalline silver product
was more efficacious in treating contact dermatitis than the
commercially available products.

Example 11

Preparation of Gels

No. 1

A commercial carboxymethyl cellulose/pectin gel (DuoDERM.TM., ConvaTec
Canada, 555, Dr. Frederik Philips, Suite 110, St-Laurent, Quebec, H4M
2.times.4) was combined with nanocrystalline silver powder prepared as
set forth in Example 3 to produce a gel with 0.1% silver. A
logarithmic reduction test was performed as follows in the gel using
Pseudomonas aeruginosa. The inoculum was prepared by placing 1
bacteriologic loopful of the organism in 5 mL of trypticase soy broth
and incubating it for 3-4 h. The inoculum (0.1 mL) was then added to
0.1 mL of gel and vortexed (triplicate samples). The mixture was
incubated for one-half hour. Then 1.8 mL of sodium thioglycollate-
saline (STS) solution was added to the test tube and vortexed. Serial
dilutions were prepared on 10.sup.-1 to 10.sup.-7. A 0.1 mL aliquot of
each dilution was plated in duplicate into Petri plates containing
Mueller-Hinton agar. The plates were incubated for 48 h and then
colonies were counted. Surviving members of organisms were determined
and the logarithmic reduction compared to the initial inoculum was
calculated. The logarithmic reduction for this mixture was 6.2,
indicating a significant bactericidal effect.

No. 2

Carboxymethyl cellulose (CMC) fibers were coated directly to produce
an atomic disordered nanocrystalline silver coating, using magnetron
sputtering conditions similar to those set forth in Example 1. The CMC
was then gelled in water by adding 2.9 g to 100 mL volume. This
material was tested using the method of No. 1. The material generated
a 5.2 logarithmic reduction of Pseudomonas aeruginosa, demonstrating
that the gel had a significant bactericidal effect.

No. 3

An alginate fibrous substrate was directly coated with an atomic
disordered nanocrystalline silver coating using magnetron sputtering
conditions similar to those set forth in Example 1. The alginate (5.7
g) was added to 100 mL volume of water to create a gel. This material
was tested using the method of No. 1. The material generated a 5.2
logarithmic reduction of Pseudomonas aeruginosa, demonstrating that
the gel had a significant bactericidal effect.

No. 4

A commercial gel containing CMC and alginate (Purilin gel, Coloplast)
was mixed with a atomic disordered nanocrystalline silver powder to
give a product with 0.1% silver. This was tested as above with both
Pseudomonas aeruginosa and Staphylococcus aureus. Zone of inhibition
data was also generated for this gel as follows. An inoculum
(Pseudomonas aeruginosa and Staphylococcus aureus)was prepared as in
No. 1 and 0.1 mL of this was spread onto the surface of Mueller-Hinton
agar in a Petri dish. A six mm hole was then cut into the agar at the
center of the Petri dish and removed. The well was filled with either
0.1 mL of the silver containing gel, a mupirocin containing cream or a
mupirocin containing ointment. The Petri plates were then incubated
for 24 h and the diameter of the zone of inhibition was measured and
recorded.

The silver containing gel produced 9 mm zone of inhibition against
both Pseudomonas aeruginosa and Staphylococcus aureus, while the
mupirocin cream and ointment produced 42 and 48 mm zones against
Staphylococcus aureus and 0 mm zones against Pseudomonas aeruginosa.

The silver containing gel reduced the Pseudomonas aeruginosa and
Staphylococcus aureus properties by 4.4 and 0.6 log reductions,
respectively, showing good bactericidal activity. The mupirocin cream
and ointment generated 0.4 and 0.8, and 0.8 and 1.6, log reductions
against Staphylococcus aureus and Pseudiomonas aeruginosa,
respectively. The silver gel had both a greater bactericidal effect
and spectrum of activity than the mupirocin containing products.

Nos. 5-10

The formula for Nos. 5-10 are summarized in the following table. Zones
of inhibitions were determined as in No. 4 and log reductions were
determined as in No. 1.

All formulae provided a broader spectrum of activity and a greater
bactericidal effect than did mupirocin in a cream or ointment form.
The mupirocin cream produced zones of inhibition of 42 and 0, and log
reduction of 0.4 and 0.8, against Staphylococcus aureus and
Pseudomonas aeruginosa, respectively.

TABLE-US-00007 Log Log Ag CZOI CZOI red'n red'n CMC PVA Powder Beta-
Methyl Propyl S. P. S. P. # (%) (%) (%) glucan paraben paraben aureus
aeruginosa aureus aeruginosa 5 2 0.1 11 13 1.4 >6 6 2 0.5 0.1 0.1 0.02
14 15 3.3 >6 7 2 0.5 0.1 13 14 2 N/A 8 2 0.5 0.1 0.1 14 14 2 N/A 9 2
0.5 0.1 0.20 14 14 2 N/A 10 2 0.5 0.1 0.5 0.1 0.20 14 14 2 >6

No. 11

A commercially available gel (glyceryl polymethacrylate) was blended
with nanocrystalline silver powder to produce a gel with a silver
content of 0.1%. This gel was tested as in Nos. 5-10 and was found to
produce zones of 15 mm against both Staphylococcus aureus and
Pseudomonas aeruginosa. Log reductions of 1.7 and >5 were produced
against Staphylococcus aureus and Pseudomonas aeruginosa. This gel
product had a greater spectrum of activity than did mupirocin cream or
ointment.

Example 12

Treatment of Adult Acne with Nanocrystalline Silver Gel Occluded by a
Hydrocolloid Dressing

A 49 year old white male experienced occasional acne vulgaris. He had
painful, raised, red papules and pustules on his shoulders. The
patient was treated with gel formulation No. 5 as set forth in Example
11. Gel formulation No. 5 was applied to the problem area of the
patient's shoulders and then occluded by a thin hydrocolloid dressing
(Craig Medical Products Ltd., Clay Gate House 46 Albert Rd. North
Reigate, Surrey, United Kingdom). The dressing remained in place for
24 hours. Upon removal the pustule was no longer painful, red, or
raised.

Treatment of Mucosal or Serosal Conditions

Example 1

Preparation of Nanocrystalline Silver Coatings on Dressings

This example shows the preparation of a bilayer nanocrystalline silver
coating on a dressing material. A high density polyethylene dressing,
DELNET.TM. or CONFORMANT 2.TM. was coated with a silver base layer and
a silver/oxide top layer to generate a coloured antimicrobial coating
having indicator value as described in Example 1 of the Treatment of
Hyperproliferative Skin conditions examples. The coating layers were
formed by magnetron sputtering under the conditions set out in the
following table.

Example 2

Preparation of Atomic Disordered Nanocrystalline Silver Powders

Atomically disordered, nanocrystalline silver powder was prepared as
described in Example 3 in the Treatment of Inflammatory Skin
conditions examples above.

Example 3

Silver solutions were prepared by immersing AgHDPE mesh from dressings
prepared as in Example 1 in reverse osmosis water that had been
pretreated with CO.sub.2 in order to reduce the pH. Two different
concentrations of silver solutions were prepared by this method, the
concentrations being 85 .mu.g/ml, and 318 .mu.g/ml. Solutions of
silver nitrate were also prepared to use as comparisons in the
experiments. The concentrations of the silver nitrate were 103 ppm of
silver and 295 ppm of silver as determined by Atomic Absorption
Spectroscopy.

The solutions were in turn placed in an ultrasonic nebulizer that
created small droplets containing dissolved and suspended parts of the
silver solution. The output from the nebulizer was directed into a
chamber made from a stainless steel frame and base. Petri dishes
containing Mueller Hinton agar streaked with 4 hold cultures of
Pseudomonas aeruginosa and Staphylococcus aureus, were exposed to the
silver solution aerosols and the silver nitrate aerosols.

The results of the tests show that silver aerosols of this invention
transmit the antimicrobial activity of the dressings to remote sites,
and such aerosols are more effective than comparable concentrations of
silver nitrate.

In many instances the delivery of antimicrobial materials may most
expeditiously be accomplished by using aerosols (e.g. treatment of
pneumonia). The drawback of aerosols is the requirement for a high
concentration of the active ingredient to ensure that a sufficient
amount is delivered to achieve the biological effect desired without
causing problems with the carrier solvent (e.g. water). It is
preferably that the equipment for producing an aerosol that contains
the dissolved and suspended components of nanocrystalline silver form
droplets of aerosol direct from the liquid form, and the aerosol
droplets must be small enough to reach the lungs. This means the
droplets should be less than approximately 10 .mu.m. To meet these
requirements the aerosol is not created by first evaporating the
liquid then condensing it to form droplets. Rather, aerosols are
generated by 1) mechanical disruption of the liquid, or 2) air under
pressure passing through some form of orifice that combines the air
and the liquid in a way that creates droplets instead of evaporating
the liquid.

Several experiments were carried out with silver solutions of this
invention and silver nitrate solutions to determine if the
antimicrobial activity of the dressing could be transferred through a
direct droplet aerosol to a Petri dish.

a) Methods

i) Equipment

The method used for the current tests was the mechanical method in the
form of an ultrasonic nebulizer. For convenience an ultrasonic
humidifier was used. The liquid containing the dissolved and suspended
silver from the dressing of Example 1 was placed in the water
reservoir of the humidifier. When power was applied to the humidifier
aerosol droplets of dissolved and suspended silver were generated and
flowed from the output nozzle.

A test chamber was constructed using a stainless steel frame with a
transparent plastic covering. The frame was placed on a stainless
steel base plate. The output nozzle from the humidifier was modified
so that the aerosol could be directed into the chamber at a height of
approximately 30 cm from the base. The plates and other test samples
were placed on the stainless steel plate and exposed to the aerosol
for a prescribed length of time.

ii) Solutions

Solution 1--A silver containing solution was prepared by immersing 518
sq. inches of the dressing from Example 1 in 1 L of reverse osmosis
water, which was treated with CO.sub.2 to maintain a pH of 6.5. After
20 minutes the concentration of silver in the water was 85 .mu.g/ml.

Solution 2--A solution containing 370 .mu.g/ml of silver from a
dressing from Example 1 was prepared as follows: 1 L of reverse
osmosis water was purged with commercial grade carbon dioxide until
the pH was 4.3.

Sufficient dressing was added to bring the pH up to 6.5. At that time,
the silver concentration was 370 .mu.g/ml.

Solution 3--Ag as AgNO.sub.3 was prepared by dissolving 0.157 g of
AgNO.sub.3 into 1 L of reverse osmosis water and mixed until
dissolved. The solution was analyzed by Atomic Absorption Spectroscopy
and found to be 102.9 ppm of silver.

Solution 4--Ag as AgNO.sub.3 was prepared by dissolving 0.427 g of
AgNO.sub.3 into 1 L of reverse osmosis water and mixed until
dissolved. The solution was analyzed by Atomic Absorption Spectroscopy
and found to be 295 ppm of silver.

iii) Aerosolization

Petri dishes, containing Mueller Hinton agar, were streaked with 4
hold cultures of Pseudomonas aeruginosa or Staphylococcus aureus. The
plates were then weighed and their exposed outer surfaces were coated
with Parafilm to prevent condensation from occurring on these
surfaces. These plates were placed in the aerosol chamber uncovered.
The ultrasonic nebulizer was then started and run for 53 minutes. The
plates were then removed from the chamber, the plastic was removed and
the dishes re-weighed so that the amount of moisture loss/gain could
be determined.

The plates were then placed in a 35.degree. C. incubator for 16 h.
After incubation the pattern and amount of growth was assessed on the
plates for both organisms.

iv) Viability Assessment

Three of the six plates made for each organism were tested to
determine if the antimicrobial effect was cidal or static in nature.
This was accomplished by rinsing or placing a piece of the clear
section of agar in the Petri dish plates into Tryptic soy broth in a
test tube and incubating for 4 h or 16 h. If the medium turned turbid
in 4 h it would indicate that the antimicrobial affect was
bacteriostatic in nature. If the organisms took more than 16 h to
grow, as indicated by turbidity, it was considered an indication that
both static and cidal effects occurred. If no growth occurred the
effect was bactericidal.

v) Results--The results are summarized in the following table.

TABLE-US-00008 Silver from Dressing AgNO.sub.3 P. P. Organism
aeruginosa S. aureus aeruginosa S. aureus Solutions 1 and 3 Ag
concentration 85 85 99 99 (.mu.g/ml) pH of test solution 6.5 6.5 About
about 6.5 6.5 Exposure Time 53 53 53 53 (minutes) Exposed are (sq.
in.) 9.8 9.8 9.8 9.8 Exp 0.8 0.8 1.05 1.05 Weight Gain (g) Growth at
16 h 0 0 0 0 (0-++++) at 48 h 0 ++ 0 ++++ Viable 4 h No No No No 16 h
No No No N/A Solutions 2 and 4 Ag concentration 370 370 300 300 (.mu.g/
ml) pH of test solution 6.5 6.5 About about 6.3 6.3 Exposure Time 53
53 53 53 (minutes) Exposed are (sq. in.) 9.8 9.8 9.8 9.8 Exp 1.14 1.14
1.12 1.12 Weight Gain (g) Growth at 16 h 0 0 0 0 (0-++++) at 48 h 0 0
0 +++ Viable 4 h No No No No 16 h No No No N/A

vi) Discussion

At the low concentration of silver in solution, the dressing generated
silver was effective in controlling the growth of both organisms while
the silver nitrate only prevented the growth of P. aeruginosa.
Viability tests showed that at the low concentration, neither form of
silver was completely bacteriocidal although the poor growth on the
dressing aerosol treated plates compared to the silver nitrate treated
plates suggests that a significant log reduction occurred in the
dressing aerosol treated plates.

At a higher concentration of silver, both dressing generated silver
(370 .mu.g/ml) and AgNO.sub.3 (300 .mu.g/ml) were effective at
controlling P. aeruginosa. Since no re-growth occurred, it is assumed
that the agent at this concentration were bactericidal. Silver
generated from the dressing was more effective than AgNO.sub.3 at
controlling S. aureus. No re-growth occurred on any plates or in the
broth indicating a total kill of the organism while in the AgNO.sub.3
treatment, a large number of organisms grew at 16 h.

Based on weight gain during aerosol treatments a dose per unit area
can be calculated. In each case for solution 1 the dose was 8.5 .mu.g/
sq. inch while for solution 2 the dose was 38 .mu.g/sq. inch. These
doses, on a per lung basis, would be less than 10 mg of silver per
hour of treatment. Each hour of treatment with dressing generated
silver aerosols appears to provide at least 48 h of protection.
Therefore the dose per day, from the high concentration treatment,
would be about 5 mg or a little less than the silver released by 2 sq.
inches of SSD per day.

A most significant advantage of using dressing generated silver may be
the lack of a toxic cation such as NO.sub.3 or sulfadiazine.

Overall, the example demonstrated that the dressing generated aerosols
are operative to transmit the antimicrobial activity of the dressings
to remote sites. Furthermore, the dressing generated aerosols were
more effective than comparable concentrations of silver nitrate.

Example 4

Aerosolized Silver Solutions in Rats

a) Materials and Methods

i) Solutions from Atomically Disordered Silver Dressings

A solution was prepared by sparging CO.sub.2 through 400 ml of reverse
osmosis water for 30 minutes at a flow rate of 1 L/min. The beaker of
water was covered with a piece of parafilm to assist in maintaining a
saturated CO.sub.2 environment. This process resulted in the pH of the
water dropping to about 4. At this point, approximately 600 square
inches of silver-coated net (AgHDPE) prepared as in Example 1 was
added to the water and stirred for approximately 40 minutes resulting
in an elevation of the pH to approximately 6.5. The solution was then
transferred to a medical nebulizer and connected to an oxygen cylinder
with a flow rate of 10 L/min. The outflow of the nebulizer was
connected to a sealed animal chamber housing the rats to be dosed.
Only the "test" rats (15 randomly assigned animals) received the
dosing. The rats received two, one-hour aerosol administrations of the
solution on the day of infection. Thereafter, the test rats were dosed
three times per day for an additional three days.

ii) Animals

Thirty male Sprague-Dawley rats were obtained from the University of
Calgary, Alberta, Canada breeding colony. These animals were specific-
pathogen free and weighed approximately 300 g. The animals were housed
in groups of 5 in plastic cages with wire mesh tops. The rats had
access to fresh water and rat chow ad libitum. All animals were
maintained in an appropriate facility with 12-hour light/dark cycles
and constant temperature and humidity, according to facility standard
operating procedures. The protocol was approved by the University of
Calgary Animal Care Committee and was conducted in accordance with
guidelines established by the Canadian Council on Animal Care.

iii) Bacteria

The bacteria used for infection of these animals were Pseudomonas
aeruginosa strain 579. The dose was previously titrated to ascertain
that a dose of up to 10.sup.10 CFU was not lethal for the animals. The
bacteria were grown overnight in Tryptic soy broth, washed once in
sterile PBS, and re-suspended in a 1/10 volume of sterile PBS.

iv) Infection

The rats were anesthetized by inhalation of 2% halothane. A 50
microliter volume of bacterial suspension was intratracheally
administered into the bronchi of each rat. This was performed non-
surgically on intubated animals. The infection process resulted in the
instillation of approximately 2.times.10.sup.9 CFU into the lungs of
each animal.

v) Sampling

On each day, a number of animals were sacrificed. The lungs of the
animals were aseptically removed, homogenized, and plated to determine
bacterial levels. A few animals were also subjected to bronchoalveolar
lavage prior to removal of the lungs. In several cases, lung
homogenates and/or lavage fluids were reserved for silver analysis.

After the first batch of the silver solution was prepared, total
silver analysis indicated that there was about 225 ppm of total silver
in the solution. The solution was reserved for several hours until
after next dosing of the animals. A second silver analysis indicated
that the total silver in solution had dropped to about 166 ppm. The
reason for the drop was immediately apparent as the silver had visibly
precipitated out of solution and had deposited on the surface of the
nebulizer. One other batch of freshly prepared solution had a total
silver concentration of 337 ppm. Regardless of the actual numbers, the
process of generating the silver solution results in a significant
quantity of silver in the solution that is aerosolized into the dosing
chamber.

The dosing chamber is not perfect. Although significant amounts of
mist are generated into the chamber, the rates tend to lie on top of
one another and are probably exposed to vastly different levels of the
silver mist.

vi) Results

i) Pulmonary Bacterial Levels

TABLE-US-00009 Day Log CFU/Test Lung Log CFU/Control Lung 1 6.2 7.3 2
4.1 7.8 3 0 6.2 4 3.5 4.8

The bacteriological results gathered from the lungs of the treated and
control animals demonstrated a sharper decline in the numbers of
bacteria present in the lungs following treatment with silver mist as
compared to controls. The results indicated that, in spite of the
small sample sizes and inconsistent exposures, a difference could
still be noted. There was considerable variation in the numbers of
bacteria recovered from individual animals within each treatment group
and, therefore, there was no significant difference in the results.
Gross examination of excised lungs suggested that there may have been
less apparent damage to the lungs in the animals treated with the
silver mist as compared to the untreated, infected animal. This was
very encouraging given the potential anti-inflammatory effects of the
nanocrystalline silver technology.

ii) Pulmonary Silver Levels

TABLE-US-00010 Sacrifice Date Rat Description Total Silver Level
Average 36999 Silver mist 1 0.50 ppm 36999 Silver mist 2 1.13 ppm 0.74
ppm 36999 Silver mist 3 0.58 ppm 37000 Silver mist 4 0.73 ppm 37000
Silver mist 5 0.70 ppm 0.72 ppm 37000 Control 1 0.08 ppm 37000 Control
2 0.10 ppm 0.09 ppm

The results of the silver analysis appear to indicate that the amount
of silver in the lung either plateaus or each dose of silver mist
deposits a certain amount of silver within the lung and this level is
significantly diminished prior to the next dosing of the animals.

The results of this experiment indicated that the method employed to
prepare the silver mist solution was reasonably reproducible and
yielded relatively high concentrations of silver in solution. However,
the silver was prone to precipitation and should be freshly prepared
prior to each dosing period. A lengthy period between preparation and
dosing, although resulting in a decrease in the amount of silver in
solution, did not result in a complete elimination of the silver from
the solution or even result in the silver concentration dropping to
very low levels.

The method employed for exposing the rats to the mist is also prone to
significant variation due to the piling up of the rats and the
resultant inconsistent exposure to the silver-containing mist.
However, the silver analyses suggested that a reasonably uniform dose
of silver was achieved when only a few animals were present within the
dosing chamber.

Regardless of the difficulties associated with the experiment, the
results were indicative of a therapeutic modality for pulmonary
infections. The results showed that the presence of silver mist was
effective in more rapidly clearing the bacterial load of the infected
lungs than is the host immune system alone. The apparently less severe
pathology associated with the rat lungs treated with the silver mist
showed that the treatment was effective for more than simply assisting
in the killing of invading organisms.

Example 5

Pulmonary Anti-Inflammatory Activity

A solution form nanocrystalline silver coated dressings (AgHDPE) from
Example 1 was prepared by sparging CO.sub.2 through 1000 ml of reverse
osmosis water using commercial CO.sub.2 Soda Syphon Charger. This
process resulted in the pH of the water dropping to about 4. At this
point, approximately 333 ml of the carbonated water was decanted into
a plastic bottle and 333 square inches of nanocrystalline silver-
coated net was added to the water. The nanocrystalline silver-coated
net and water were placed in 37.degree. C. shaker incubator and shaken
at 180 RPM for 30 minutes to elevate the pH to approximately 5.8. The
solution was then transferred to a beaker and stirred vigorously for 2
minutes to raise the pH to approximately at 7.3. The dissolution
solution was transferred to a commercial nebulizer which was connected
to a medical air cylinder with a flow rate of approx. 20 L/min. The
outflow of the nebulizer was connected to a animal chamber housing the
rats to be dosed. Only the "test" rats (12 randomly assigned animals)
received the dosing. The rats received two -1/2 hour aerosol
administrations of the test solution on the day of infection.
Thereafter, the test rats were dosed 3 times per day for an additional
one and a half days.

Thirty male Spragu-Dawley rats were obtained. These animals are
specific-pathogen free and weighed approximately 400 g. The animals
are housed in groups of four in plastic cages with wire mesh tops. The
rats had access to fresh water and rat chow ad libitum. All animals
were maintained in an appropriate facility standard operating
procedures.

The bacteria used for infection of these animals were Pseudomonas
aeruginosa strain 5588. The dose was previously titrated to ascertain
that a dose of up to 10.sup.9 CFU was not lethal for the animals. The
bacteria were grown overnight in Tryptic soy broth, washed once in
sterile PBS and resuspend in sterile PBS. The final concentration of
the inoculum was 4.times.10.sup.9 CFU/ml.

The rats were anesthetized by inhalation of 2% halothane. A 400
microliter volume of bacterial suspension was intratracheally
administered in the bronchi of each rat. This was performed non-
surgically on intubated animals. The infection process resulted in the
instillation of approximately 10.sup.9 CFU into the lungs of each
animal.

The three treatment groups of rats and treatment schedules were as
follows:

TABLE-US-00011 Group 1 Infected, not treated (12 Rats) Group 2
Infected, animal will be treated by intramuscularly injection of
Tobramycin at 30 mg/kg (12 mg/rat) once daily (12 Rats) Group 3
Infected and treated, using nanocrystalline silver solution and
nebulizer (Nebulized Ag), three times a day (12 Rats) Day One 10:00 AM
Infection 4:00 PM First treatment (For Group 2, Nebulized Ag for Group
3) 8:00 PM Nebulized Ag treatment for Group 3 Day Two 9:00 AM
Injection treatment for Group 2, Nebulized Ag for Group 3 1:00 PM
Sacrifice and sample six Rats in each group 3:00 PM Nebulized Ag
treatment for Group 3 8:00 PM Nebulized Ag treatment for Group 3 Day
Three 9:00 AM Injection treatment for Group 2, Nebulized Ag for Group
3 1:00 PM Sacrifice and sample six Rats in each group

On each day, six rats of each group of animals were sacrificed. The
lungs of the animals were aseptically removed, homogenized and plated
to determine bacterial levels. Lung samples were collected for
histological examination. Three lung homogenates were reserved for
silver analysis. Lungs were grossly scored (absent=0, mild=1,
moderate=2, and severe=3) based on the degree and involvement of
consolidation, hemorrhage, edema and necrosis based upon gross
observation.

Histopathology was scored (0-4) based upon the degree of consolidation
and inflammation (neutrophil infiltration). The entire right middle
lobes of all rats were collected for histopathology. As whole lobes
were selected there was no bias toward any sample. All samples were
fixed in neutral buffered formalin at the time the lung was removed
from the thorax. It was fixed overnight, dehydrated and embedded in
was. Sections were obtained which were hydrated and stained with
haematoxylin and eosin.

All sections were examined by a veterinary pathologist who was blinded
to the treatment groups, until after the samples were scored and
comments were provided. The Scores and comments are provided in Table
5. (0=normal, 1=slight, 2 moderate, 3 severe, 4 very severe).

Tissue Colony Counts:

At 24 hours, there was not a significant reduction in the number of
colony forming units (cfu) in the nebulized Ag group compared to the
control but at 48 hours there was a significant reduction in the
bacterial numbers in the nebulized Ag animals. The Tobramycin treated
animals had a similar cfu counts to the controls at time 24 hours and
48 hours.

TABLE-US-00012 Control Tobramycin Nebulization 24 h animal 1 0 2 1 2 0
3 1 3 3 1 0 4 3 3 0 5 2 2 3 6 3 2 1 48 h animal 7 2 1 1 8 1 2 1 9 1 1
0 10 1 1 0 11 3 1 1 12 Dead Dead Dead

Histopathology of Lung Samples:

Both the control and the Tobramycin treated rats had similar
pathology. These are outlined in Table 6. At 24 and 48 hours severe
infiltration of polymorphonuclear leukocytes (PMN's) into the
interstitial spaces of the lung was observed. These cellular elements
could also be identified in alveolar and bronchiolar spaces but to a
lesser extent. The pulmonary vessels were dilated and the alveolar
spaces were filled with proteinaceous material. The silver-nebulized
rats had occasional infiltration of PMN's and no evidence of
accumulation of fluids in alveolar or bronchiolar spaces.

TABLE-US-00013 Histopathology of Lung Samples Removed from Rats Inflam
Consol Treatment Time Score Score Comments Control (1) 24 3 3 Severe
infiltration of PMN into interstitial spaces. Proteinaceous secretion
in alveolar spaces. Occasional PMN in alveolar and bronchiolar space.
Consolidation in affected areas. Involvement of 70% of sample.
Interstitial Pneumonia. Control (2) 24 3 3 Severe infiltration of PMNs
into interstitial spaces. Proteinaceous secretion in alveolar spaces.
Occasional PMN in alveolar and bronchiolar space. Consolidation in
affected areas. Involvement of 80% of sample. Interstitial Pneumonia
Tobramycin (1) 24 3 3 Severe infiltration of PMNs into interstitial
spaces. Proteinaceous secretion in alveolar spaces. Occasional PMN in
alveolar and bronchiolar space. Consolidation in affected areas.
Involvement of 90% of sample. Interstitial Pneumonia. Tobramycin (2)
24 3 3 Severe infiltration of PMNs into interstitial spaces.
Proteinaceous secretion in alveolar spaces. Occasional PMN in alveolar
and bronchiolar space. Consolidation in affected areas. Involvement of
80% of sample. Interstitial Pneumonia. Nebulized Ag (1) 24 0 1 No PMNs
in area. Slight consolidation. Normal Lung Nebulized Ag (2) 24 1 1
Slight infiltration of PMNs around vessels and brocheoli. Control (1)
48 3 3 Severe infiltration of PMNs into interstitial spaces.
Proteinaceous secretion in alveolar spaces. Occasional PMN in alveolar
and bronchiolar space. Consolidation in affected areas. Involvement of
80% of sample. Interstitial Pneumonia. Control (2) 48 2 2 Severe
infiltration of PMNs into interstitial spaces. Proteinaceous secretion
in alveolar spaces. Occasional PMN in alveolar and bronchiolar space.
Consolidation in affected areas. Involvement of 60% of sample.
Interstitial Pneumonia. Tobramycin (1) 48 3 3 Severe infiltration of
PMNs into interstitial spaces. Proteinaceous secretion in alveolar
spaces. Occasional PMN in alveolar and bronchiolar space.
Consolidation in affected areas. Involvement of 70% of sample.
Interstitial Pneumonia. Tobramycin (2) 48 3 3 Severe infiltration of
PMNs into interstitial spaces. Proteinaceous secretion in alveolar
spaces. Occasional PMN in alveolar and bronchiolar space.
Consolidation in affected areas. Involvement of 70% of sample.
Interstitial Pneumonia. Slight infiltration of PMNs around vessels and
brocheoli. Nebulized Ag (1) 48 1 0 Slight infiltration of PMNs around
vessels and brocheoli. Nebulized Ag (2) Normal lung.

The nebulized nanocrystalline silver reduced bacterial colonization in
Pseudomonas infected lungs reduced injury as determined by gross
pathology (consolidation, hemorrhage, edema) in Pseudomonas infected
lungs. Further, the nanocrystalline silver delivered by aerosol
reduced pulmonary inflammation (primarily PMN infiltration) in
Pseudomonas infected lungs compared to Tobramycin (IM).

Example 6

Pulmonary Anti-Inflammatory Activity

A solution was prepared by sparging CO.sub.2 through 1000 ml of
reverse osmosis water using commercial CO.sub.2 Soda Syphon Charger.
This process results in the pH of the water dropping to about 4. At
this point, approximately 333 ml of the carbonated water was decanted
into a plastic bottle and 333 square inches of nanocrystalline silver-
coated net was added to the water. The nanocrystalline silver-coated
net and water were placed in 37.degree. C. shaker incubator and shaken
at 180 RPM for 30 minutes to elevate the pH to approximately 5.8. The
solution was then transferred to a beaker and stirred vigorously for 2
minutes to raise the pH to approximately at 7.3. The solution had a
final silver concentration of approximately 400 ppm.

Test solutions of silver nitrate (400 ppm) and silver acetate (400
ppm) were prepared by dissolving the silver salts in deionized water.
A colloidal silver solution (20 ppm) in was obtained from a commercial
source.

The dissolution solutions were transferred to commercial nebulizers
which were connected to a Medical air cylinder with a flow rate of
approx. 20 L/min. The outflows of the nebulizers were connected to an
animal chamber housing the rats to be dosed. All rats (40 randomly
assigned animals) received the dosing. The rats received two -1/2 hour
aerosol administrations of the test solutions on the day of infection.
Thereafter, the test rats were dosed 3 times per day for an additional
one and a half days.

Forty male Sprague-Dawley rats were obtained. These animals are
specific-pathogen free and weighed approximately 400 g. The animals
were housed in groups of four in plastic cages with wire mesh tops.
The rats had access to fresh water and rat chow ad libitum. All
animals were maintained in an appropriate facility with 12-hour light/
dark cycles and constant temperature and humidity, according to
facility standard operating procedures.

The bacteria used for infection of 20 these animals were Pseudomonas
aeruginosa strain 5588. The dose was previously titrated to ascertain
that a dose of up to 10.sup.9 CFU was not lethal for the animals. The
bacteria were grown overnight in Tryptic soy broth, washed once in
sterile PBS and resuspend in sterile PBS. The final concentration of
the inoculum was 4.times.10.sup.9 CFU/ml.

The rats were anesthetized by inhalation of 2% halothane. A 400
microliter volume of bacterial suspension was intratracheally
administered into the bronchi of each rat. This was performed non-
surgically on intubated animals. The infection process resulted in the
instillation of approximately 10.sup.9 CFU into the lungs of each
animal. Group 1 & 2: Not infected and infected, treated with nebulized
silver nitrate. (10 Rats) Group 3 & 4: Not infected and infected,
treated with nebulized colloidal silver. (10 Rats) Group 5 & 6: Not
infected and infected, treated with nebulized nanocrystalline silver.
(10 Rats) Group 7 & 8: Not infected and infected, treated with
nebulized silver acetate. (10 Rats)

TABLE-US-00014 The treatment schedule was as follows: Day One Day Two
10:00 AM Infection 9:00 AM Third Treatment 4:00 PM First Treatment
1:00 PM Sacrifice, sample 5 rats/Gp 8:00 PM Second Treatment

All rats of each group of animals were sacrificed after 24 h. The
lungs of the animals were aseptically removed, homogenized and plated
to determine bacterial levels. Lung samples were collected for
histological examination. Three lung homogenates were reserved for
silver analysis. Lungs were grossly scored (absent=0, mild=1,
moderate=2, and severe=3) based on the degree of involvement of
consolidation, hemorrhage, edema and necrosis based upon gross
observation.

Histopathology was scored (0-4) based upon the degree of consolidation
and inflammation (neutrophil infiltration). The entire right middle
lobes of all rats were collected for histopahtology. As whole lobes
were selected there was no bias toward any sample. All samples were
fixed in neutral buffered formalin at the time the lung was removed
from the thorax. It was fixed overnight, dehydrated and embedded in
wax. Sections were obtained which were hydrated and stained with
haematoxylin and eosin.

All sections were examined by a veterinary pathologist who was blinded
to the treatment groups, until after the samples were scored and
comments were provided, with scoring being (0=normal, 1=slight,
2=moderate, 3=severe, 4=very severe).

All rats in the silver nitrate, silver acetate and colloidal silver
groups had lung that were grossly scored as moderately to severely
inflamed while the lungs of the nanocrystalline group were grossly
scored as normal to slightly inflamed. This was confirmed by the
histopathological analyses.

The nanocrystalline derived silver solution had pulmonary anti-
inflammatory properties while the other forms of silver did not.

Example 7

Treatment of an Infected Throat with a Nanocrystalline Silver Derived
Solution

A forty-nine year old male was suffering from an infected throat. The
condition was accompanied by fever and a severe pain that made
swallowing very difficult and limited the patients ability to sleep. A
solution of nanocrystalline derived silver was prepared using a method
similar to Example 1. This solution was gargled for one minute and
repeated 3 times over the next ten minutes. Within an hour the pain
was reduced to the point where the patient could sleep. The treatment
was repeated every four hours for 16 h and then once 8 h later. The
throat infection was cleared as determined by the elimination of fever
and pain. No further symptoms occurred.

Example 8

Preparation of Gels

Gels were prepared as described above in Example 11 in the Treatment
of Inflammatory Skin Conditions examples above.

Apoptosis Induction/MMP Modulation

Example 1

Preparation of Nanocrystalline Silver Coatings on Dressings

This example shows the preparation of a bilayer nanocrystalline silver
coating on a dressing material. A high density polyethylene dressing,
DELNET.TM. or CONFORMANT 2.TM. was coated with a silver base layer and
a silver/oxide top layer to generate a coloured antimicrobial coating
having indicator value as described in Example 1 of the Treatment of
Hyperproliferative Skin conditions examples. The coating layers were
formed by magnetron sputtering under the conditions set out in the
following table.

Example 2

Preparation of Atomic Disordered Nanocrystalline Silver Powders

Atomically disordered, nanocrystalline silver powders were prepared as
described in Example 3 in the Treatment of Inflammatory Skin
Conditions examples above.

Example 3

Preparation of Gels

Gels were prepared as described above in Example 11 in the Treatment
of Inflammatory Skin Conditions examples above.

Example 4

Effects of Antimicrobial Silver on Apoptosis and Matrix
Metalloproteinases in a Porcine Model

A porcine model was used to examine the effects of an antimicrobial
metal formed with atomic disorder, preferably silver, on apoptosis and
matrix metalloproteinases. Young, commercially produced, specific
pathogen free domestic swine (20-25 kg) were used in these studies.
The animals were conditioned in an animal facility for one week prior
to any experimental manipulation. Typically, three animals were used
in each experiment. The animals received water and hog ration
(Unifeed.TM., Calgary, Alberta) without antibiotics ad libitum, were
housed individually in suspended stainless steel cages (5'.times.6'),
and maintained in a controlled environment with 12 hours of light per
day. The study was approved by the University of Calgary Animal Care
Committee and was conducted in accordance with guidelines established
by the Canadian Council on Animal Care.

Antimicrobial silver metal was administered in the form of a dressing.
The dressings included:

i) AB--nanocrystalline silver-coated dressing (the non-foam, three-
layer dressing as set out in Example 1), comprising two layers of
silver-coated high density polyethylene (HDPE) bonded on either side
of an absorbent rayon/polyester core;

ii) AgHDPE--nanocrystalline silver coated HDPE layers aseptically
separated from the absorbent core of the AB dressings;

iii) Control--identical in construction to the AB dressing except that
the HDPE was not coated with nanocrystalline silver;

iv) Gauze--the absorbent rayon/polyester core of the AB dressings;

v) cHDPE--the uncoated HDPE aseptically removed from the absorbent
core of the control dressings; and

vi) SN--sterile piece of the gauze dressing to which 24 .mu.g silver/
square inch (from silver nitrate) was added. This amount of silver is
equivalent to the amount of silver released from a square inch of the
AB dressing immersed in serum over a 24 hour period, as determined by
atomic absorption analysis.

Dressings (i)-(iii) were gamma sterilized (25 kGy) prior to use. All
dressings were moistened with sterile water prior to application to
the incision. In some cases, the incisions were covered with a layer
of occlusive polyurethane (Tegaderm.TM., 3M Corp., Minneapolis,
Minn.).

Three isolates of bacteria were used in the inoculum, including
Pseudomonas aeruginosa, Fusobacterium sp., and coagulase-negative
staphylococci (CNS) (Culture Collection, University of Calgary,
Calgary, Alberta). The bacterial strains were grown under appropriate
conditions overnight prior to the day of surgery. On the morning of
surgery, the organisms were washed with sterile water and resuspended
to a final density of approximately 10.sup.7 CFU/mL. The bacteria were
mixed together in a ratio of 1:0.5:1 (Pseudomonas:CNS:Fusobacterium)
in water. Sufficient inoculum was prepared to wet the incision created
in each experiment. This procedure resulted in the incisions initially
being evenly contaminated with approximately 8.times.10.sup.4 CFU/
cm.sup.2.

Prior to treatment, animals were sedated by an intramuscular injection
of a mixture of 10 mg/kg ketamine (Ketalean.TM., MTC Pharmaceuticals,
Cambridge, ON) and 0.2 mg/kg acepromazine (Atravet.TM., Ayerst
Laboratories), followed by complete anesthesia induced by mask
inhalation of 1-2% halothane (MTC Pharmaceuticals). Following
induction of anesthesia, the dorsal and lateral thorax and abdomen of
each animal was clipped using a #40 Osler blade and the skin
subsequently scrubbed with a non-antibiotic soap, and allowed to dry
prior to incision.

Animals typically received 20 full-thickness incisions, 10 on each
side of the dorsal thorax. The incisions were created using a 2 cm
diameter trephine. An epinephrine solution was then applied to the
incisions to provide for complete hemostasis prior to inoculation. The
incisions were contaminated by covering them with gauze sponges soaked
with the bacterial inoculum. The wet sponges were covered with an
occlusive barrier and allowed to stand for 15 minutes. In some
instances, an incision was then sampled to determine the initial
inoculum. Following any requisite sampling, the incisions were dressed
with the appropriate dressings and covered with an occlusive layer
that was secured with Elastoplast.TM. tape (Smith & Nephew, Lachine,
QC). All animals received narcotic pain medication (Torbugesic.TM.,
Ayerst Laboratories, Montreal, QC, 0.2 mg/kg), as required.

The experimental and control groups are summarized in the following
table:

TABLE-US-00015 Animal # Left Side (Silver Treatment) Right Side
(Controls) Pig 1 silver nitrate (SN) on gauze gauze moistened with
water Pig 2 AgHDPE cHDPE Pig 3 AB control

A 2 cm diameter circle of the appropriate dressing was applied to each
incision. For Pig 1, incisions on the left side were dressed with
silver nitrate-moistened (SN) gauze, while control incisions on the
right side received water-moistened gauze dressing. For Pig 2, the
incisions on the left side were dressed with silver-coated HDPE
(AgHDPE), while the control incisions on the right side received non-
coated HDPE (cHDPE). For Pig 3, the incisions on the left side were
dressed with AB dressing, while incisions on the right side received
the vehicle control. For these experiments, each incision was
individually covered with an occlusive film dressing (Tegaderm.TM., 3M
Corp., Minneapolis, Minn.).

Each day following incision (up to 5 days), the dressing materials
from each of the experimental and control groups were collected and
pooled within each group. These dressing materials were then placed in
conical centrifuge tube containing glass wool. The tubes and contents
were centrifuged to remove all liquid from the dressings. The glass
wool was then placed into a 5-mL syringe and pressed to recover the
incision fluid from each of the six sample sets. The incisions were
rebandaged in an identical manner to the original dressing format each
time. Incision fluid which collected under the occlusive dressing was
also aspirated and reserved for analysis. Due to the small volumes
collected from each incision, it was necessary to pool the collected
fluid from each of 10 incisions dressed with the same type of
dressing. All incision fluids were stored at -80.degree. C. until
analysis.

Prior to enzyme zymography or activity assays, the protein
concentrations of the incision fluid samples were compared to ensure
that the protein levels in each sample were similar. The samples were
diluted 1:100 in water and assayed using the BCA Protein Assay
SysteM.TM. (Pierce Chemical, Rockford, Ill.). A standard curve was
concurrently constructed using dilutions of bovine serum albumin.
Incision fluid was diluted in water and then mixed with an equal
volume of sample buffer (0.06 M Tris-HCl, pH 6.8; 12% SDS; 10%
glycerol; 0.005% bromophenol blue). The samples were then
electrophoresed on 10% polyacrylamide (BioRad, Mississauga, ON) gels
containing 0.1% gelatin. The proteins were then incubated in
renaturing buffer (2.5% Triton.TM. X-100) for 90 minutes at 37.degree.
C. Following enzyme renaturation, the gels were incubated overnight in
substrate buffer (50 mM Tri-HCl, pH 7.8; 5 mM CaCl.sub.2; 200 mM NaCl;
0.02% Brij-35) with or without 10 mM 1,10 phenanthroline. The gels
were subsequently stained with a standard Coomassie Blue stain and
destained in methanol/acetic acid. Unless otherwise indicated, all
chemicals were obtained from Sigma-Aldrich (Oakville, ON).

The incision fluid samples were assayed for the total amount of
protein present. These values were between 30-80 mg/mL. The samples
from individual animals were even more similar, varying by only 10-20
mg/mL in the pooled incision fluid.

i) Assay for Activity of Total MMPs

The total MMP activity of the incision fluid samples was determined by
incubating diluted incision fluid with a quenched fluorescein-
conjugated substrate (EnzChek DQ gelatin.TM., Molecular Probes,
Eugene, Oreg.) for approximately 20 hours. Following incubation, the
samples were read in a fluorometer (excitation 1=480 nm; emission
1=520 nm). Activity was compared to a collagenase standard as well as
experimental versus control incision fluids.

FIG. 4 shows the change in total MMP activity from differently treated
incision sites over a five-day period. The silver-coated HDPE (AgHDPE)
results were essentially identical to those obtained using the silver-
coated dressing (AB). Similarly, the gauze, non-coated HDPE (cHDPE),
and control dressings yielded results essentially identical to each
other and to untreated incisions under occlusion from which incision
fluid was collected. Only the results from the control, silver-coated
dressing (AB), silver-coated HDPE (AgHDPE), and silver nitrate
moistened gauze (SN) are thus shown. The total MMP activity of the
incision fluid sample from the control dressing was low for the first
few days, then rose dramatically and remained high for the duration of
the experiment. Similarly, the silver-nitrate moistened gauze (SN)
demonstrated an almost identical pattern of total MMP activity.
Results from the silver-coated dressing (AB) yielded dramatically
different results. The level of MMP activity remained steady for the
duration of the experiment and did not spike to high levels. Instead,
it remained at a level roughly 60% lower than the highest level of
activity reached in control or silver-nitrate moistened gauze (SN).

ii) Assay for Activity of Gelatinases

Gelatinases include MMP-2 (secreted by fibroblasts and a wide variety
of other cell types) and MMP-9 (released by mononuclear phagocytes,
neutrophils, corneal epithelial cells, tumor cells, cytotrophoblasts
and keratinocytes). The gelatinases degrade gelatins (denatured
collagens) and collagen type IV (basement membrane). Zymograms were
run to examine changes in the levels and activity of MMP-9 and MMP-2
over the duration of the experiment.

Results of the zymograms for the control and silver nitrate moistened
gauze (SN) appeared to be identical. The levels of MMP-9 declined over
the period examined, particularly levels of the active form of MMP-9.
The silver-coated dressing (AB) demonstrated higher levels of active
MMP-9 than for the control. On Day 2, the silver-coated dressing (AB)
showed lower levels of active MMP-9 than in the control. On Day 4, the
silver-coated dressing (AB) showed little active MMP-9. In the
control, the amount of the latent enzyme appeared to decrease while
the active form of MMP-9 increased, particularly up to Day 4.

There was not much difference in the levels of MMP-2 activity for the
silver-coated dressing (AB) over the duration of the experiment.
However, there was an increase in the level of active MMP-2 in the
control dressing by Day 5. It was also observed that the levels of
some other, unidentified, gelatinolytic enzymes also decreased in the
silver-coated dressing (AB) compared to the control.

iii) Assay of Total Protease Activity

Since MMPs have proteolytic activity, the total protease activity in
incision fluid samples was assessed by incubating the samples with 3
mg/mL azocasein in 0.05 M Tris-HCl, pH 7.5 for 24 hours at 37.degree.
C. The undigested substrate was then precipitated by the addition of
20% trichloroacetic acid. The absorbance of the supernatant was then
assessed using a spectrophotometer, 1=400 nm. The absorbance was
compared to a standard curve prepared with bovine pancreatic trypsin.

Results paralleled those obtained in the total MMP assay above. The
incision fluid samples for the control and silver nitrate moistened
gauze (SN) demonstrated a pronounced increase in activity after Day 2
(FIG. 5). Incision fluid from the silver nitrate moistened gauze (SN)
also demonstrated a marked increase in the total protease activity
compared to control dressing incision fluid (FIG. 4). However, the
total protease activity in the incision fluids of the silver coated
dressings (AB) remained constant over the duration of the experiment.

Antimicrobial silver was thus demonstrated to be effective in
modulating overall MMP activity. However, silver nitrate was not
effective in modulating MMP activity in spite of the Ag.sup.+
concentration being approximately the same levels as were expected to
be released from the silver-coated dressing (AB) over the same period
of time (24 h) between applications. The reason for the difference in
effects may be related to the inherent nature of the two silver
formulations. In the case of silver nitrate, although the silver was
added to provide a similar final concentration of Ag.sup.+ as was
anticipated to be released from the silver-coated dressing (AB), the
Ag.sup.+ ions were added at once. It would thus be expected that the
serum proteins and chlorides within the incision fluid would quickly
inactivate a large portion of the Ag.sup.+. In the case of the silver-
coated dressing (AB), the silver is continuously released to maintain
a steady-state equilibrium, maintaining an effective level of silver
in the incision for a prolonged period.

iv) Apoptosis

Histological assessment of cell apoptosis was carried out in order to
determine whether the silver-coated dressing (AB) affected apoptosis
within the incision.

Histological Observations of Porcine Tissue

Samples of tissue from the incisions were collected daily for the
duration of the experiment, except for Day 1, and examined for
evidence of apoptosis. The samples were fixed in 3.7% formaldehyde in
PBS for 24 hours, then embedded in paraffin, and cut into 5 mm thick
sections. The samples were then de-waxed with Clearing Solvent.TM.
(Stephan's Scientific, Riverdale, N.J.) and rehydrated through an
ethanol:water dilution series. The sections were treated with 20 mg/mL
proteinase K (Qiagen, Germantown, Md.) in 10 mM Tris-HCl (pH 7.4) for
30 minutes at room temperature.

Terminal deoxynucleotidyl transferase nick end labeling (TUNEL
staining) was performed using an In Situ Cell Death Detection POD
Kit.TM. (Boehringer Mannheim, Indianapolis, Ind.). Using this
technique, cells which stain brown are those being eliminated by
apoptosis. Endogenous peroxidase was blocked with 3% hydrogen peroxide
in methanol for 10 minutes at room temperature then cells were
permeabilized with 0.1% Triton.TM. X100 (in 0.1% sodium citrate) for 2
minutes on ice. After permeabilization, the samples were treated with
the terminal transferase enzyme solution incubated in a humidified
chamber at 37.degree. C. for 60 minutes. Following labelling, the
samples were washed once with 1.0% Triton.TM. X-100 and twice with
PBS. The sections were incubated with Converter-POD.TM. (Boehringer
Mannheim, Indianapolis, Ind.) in a humidified chamber at 37.degree. C.
for 30 minutes, and repeated washing with 1.0% Triton.TM. X-100 and
PBS. Subsequently, the samples were incubated with DAB substrate
(Vector Laboratory Inc., Burlingame, Calif.) for 10 minutes at room
temperature and washed with 1.0% Triton.TM. X-100 and PBS. It was also
necessary to counterstain the sections with hematoxylin nuclear
counterstain (Vector Laboratory Inc., Burlingame, Calif.) for 10
seconds.

The prepared samples were then ready to be observed by light
microscopy for evidence of apoptosis. For a positive control, the
permeabilized sections were treated with 100 mg/mL DNase I in PBS for
10 minutes at room temperature to induce DNA strand breaks. For
negative controls, the terminal transferase enzyme, POD or DAB were
omitted between each labelling step.

In all samples examined, there was little difference between the
control and silver nitrate moistened gauze (SN). However, significant
apoptosis of the cell population was observed in incisions of the
silver-coated dressing (AB). In the control incision, there were
significant numbers of polymorphonuclear leukocytes (PMNs) and few
fibroblasts, while in incisions of the silver-coated dressing (AB),
there were significantly more fibroblasts and few PMNs.

Histopathological Scoring of Porcine Tissue

Animals were anesthetized as described above of Days 1, 4, and 7. A
mid-incision biopsy was collected with a sterile 4 mm biopsy punch.
The tissue was fixed in 10% neutral buffered formalin, embedded in
methacrylate and sectioned (2-5 mm thick). The sections were stained
with Lee's methylene blue and basic fuschin to demonstrate the
cellular organization and bacteria. A pathologist blinded to the
treatments scored the sections based on the presence of fibroblasts,
PMNs and bacteria as follows: 0=absent; +=occasional with 1-5 per high
power field of view; ++=moderate with 6-20 per high power field of
view; +++=-abundant with 21-50 per high power field of view; ++++=very
abundant with more than 50 per high power field of view (see the
following table).

TABLE-US-00016 Day Post- Incision Dressing Fibroblasts PMNs Bacteria 1
Silver coated ++ ++ + (AB) 1 Control 0 +++ ++++ 4 Silver coated ++++ +
+ 0 (AB) 4 Control + ++++ ++++ 7 Silver Coated ++++ + 0 (AB) 7 Control
+++ +++ +++

The microscopic observation of the biopsy samples revealed that the
infiltrating cell types were significantly different between the
control and silver-coated dressings (AB). The control incisions were
characterized by a large numbers of PMNs, while the silver-coated
dressings (AB) demonstrated a larger proportion of fibroblasts and
monocytes. The relative abundance of the fibroblasts in incisions of
the silver-coated dressings (AB) became increasingly pronounced
through to Day 7, as compared to the control incisions that remained
populated largely by PMNs and monocytes. The staining method enabled
staining also of bacteria, which was abundant in the control incision
but generally absent in the incisions of the silver-coated dressings
(AB).

Incisions treated with the nanocrystalline antimicrobial silver thus
demonstrated more extensive apoptosis than did cells from incisions
treated with either control or silver nitrate dressings. During the
first two days following incision, the cell type which demonstrated
the most pronounced increase in apoptosis were neutrophils. This
suggests that part of the reason for the moderated neutrophil presence
and the resultant modulation of MMP levels was due to neutrophil
apoptosis. It has been shown that the number of apoptotic cells
increases as the incision closes and that this is part of the
mechanism involved in the decrease in cellularity of the maturing scar
tissue (Desmouliere, A., Badid, C., Bochaton-Piallat, M. and Gabbiani,
G. (1997) Apoptosis during wound healing, fibrocontractive diseases
and vascular wall injury. Int. J. Biochem. Cell Biol. 29: 19-30.). The
results suggest that the maturing of the nascent dermal and epidermal
tissues may also be accelerated in the presence of the nanocrystalline
antimicrobial metals. The findings indicated that acceleration in
healing induced by the nanocrystalline antimicrobial metals is
associated with a reduction of local MMP activity, as well as with an
increased incidence of cell apoptosis within the incision.

Example 5

Clinical Study on the Effect of Silver-Coated Dressings on MMPs and
Cytokines

This study was conducted to assess the effect of the silver-coated
dressing on the concentrations of MMPs and cytokines in non-healing
wounds over time during treatment. The modulation of the levels of
active MMPs and cytokines may alleviate the inflammatory response in a
wound, allowing the wound to advance through the subsequent stages of
wound healing culminating in a healed wound.

A total of 10 patients with non-healing venous stasis ulcers were
randomly assigned to treatment with a silver-coated dressing (5
patients) or a control dressing (5 patients). The silver-coated
dressing was prepared as in Example 1. The control dressing was
identical in construction to the silver-coated dressing of Example 1,
except that the HDPE was not coated with silver. The ulcers were
dressed in appropriate pressure dressings to correct the underlying
medical problem. Samples of the ulcer fluid were collected before
treatment (day 0) and at weekly intervals (days 1, 7, 14 and 21) by
removing the silver-coated dressing or control dressing, and replacing
the dressing with Tegaderm.TM. occlusive dressing (3M Corp.,
Minneapolis, Minn.) for one hour to allow wound fluids to collect. The
fluid samples were aspirated from below the dressing in a syringe, and
were frozen at -80.degree. C. until assayed.

Assays were conducted for active MMP-9, active MMP-2, Tumor necrosis
factor-.alpha. (TNF-.alpha.) and Interleukin-1.beta. (IL-1.beta.).
High levels of MMP-9 and MMP-2 are predominant in non-healing wounds,
with levels decreasing over time in normal healing wounds. Released by
activated macrophages, TNF-.alpha. and IL-1.beta. are indicators of
wound inflammation. Levels of TNF-.alpha. and IL-1.beta. are elevated
in non-healing wounds and increase release of pro-MMPs, for example,
MMP-9 and MMP-2.

To measure the levels of active MMP-9 and MMP-2, enzyme capture assays
(BioTrak, N.J.) were conducted. In this method, active enzyme is
detected through activation of a modified pro-detection enzyme and the
cleavage of its chromogenic peptide substrate. The resultant color is
read by spectrophotometer, and the concentration of MMP is determined
by interpolation of a standard curve, expressed in ng/ml (see results
in FIGS. 6 and 7).

To assay the levels of cytokines, IL-1.beta. levels were measured
using a sandwich immunoassay (BioTrak, N.J.), while TNF-.alpha. levels
were measured by a high sensitivity sandwich antibody assay (BioTrak,
N.J.). In both methods, endogenous cytokine is bound to an immobilized
antibody and then detected by an addition of a biotinylated antibody,
followed by a colorimetric substrate. The color is measured by a
spectrophotometer, and the concentrations of TNF-.alpha. and
IL-1.beta. are determined by interpolation of a standard curve and
expressed as pg/ml (see results in FIGS. 8 and 9).

Total protein levels were measured for each sample to standardize the
measures of the MMPs and cytokines. Total protein levels were measured
using BCA Protein Assay System.TM. (Pierce Chemical, Rockford, Ill.).
No protein level of any sample was significantly different from the
total mean.

FIG. 6 is a graph showing the concentrations (ng/ml) of active MMP-9
in fluid samples recovered from ulcers dressed with silver-coated
dressing (Silver) and control dressing (Control) at days 0, 1, 7, 14
and 21. The levels of active MMP-9 decreased to a normal level, and
were suppressed over time with the silver-coated dressing compared to
the control dressing, demonstrating a modulating effect of the silver-
coated dressing.

FIG. 7 is a graph showing the concentrations (ng/ml) of active MMP-2
in fluid samples recovered from ulcers dressed with silver-coated
dressing (Silver) and control dressing (Control) at days 0, 1, 7, 14
and 21. The levels of active MMP-2 were not significantly different
with the silver-coated dressing and the control dressing.

FIG. 8 is a graph showing the concentrations (pg/ml) of TNF-.alpha. in
fluid samples recovered from ulcers dressed with silver-coated
dressing (Silver) and control dressing (Control) at days 0, 1, 7, 14
and 21. The levels of TNF-.alpha. were suppressed over the treatment
period, and did not increase significantly over the treatment period
with the silver-coated dressing, while the levels in the control
dressing increased, demonstrating a modulating effect of the silver-
coated dressing.

FIG. 9 is a graph showing the concentrations (pg/ml) of IL-1.beta. in
fluid samples recovered from ulcers dressed with silver-coated
dressing (Silver) and control dressing (Control) at days 0, 1, 7, 14
and 21. The levels of IL-1.beta. were not significantly different with
the silver-coated dressing and the control dressing.

The study suggests that the modulation of the MMP-9 and TNF-.alpha.
levels is responsible for improved wound healing and reduced
inflammation with silver-coated dressings. In comparison, the levels
of MMPs and cytokines did not decrease over time with the control
dressings.

This example and Example 4 above, taken together with the evidence
that the silver materials herein disclosed are capable of reducing
inflammation (see co-pending

U.S. patent application Ser. Nos. 10/131,568 filed on Apr. 23, 2002;
10/131,511 filed on Apr. 23, 2002; 10/131,509 filed on Apr. 23, 2002;
10/131,513 filed on Apr. 23, 2002; demonstrates a method of reducing
inflammation in a patient in need thereof, by contacting an area of
inflammation or an inflammatory cell with a therapeutically effective
amount of the antimicrobial metals in a crystalline form. The
antimicrobial metals are characterized by sufficient atomic disorder,
such that the metal, in contact with an alcohol or water-based
electrolyte, releases atoms, ions, molecules, or clusters of at least
one antimicrobial metal at a concentration sufficient to modulate the
release of one or both of MMP-9 and TNF-.alpha.. Excessive TNF
production has been reported in diseases, such as cancer and
autoimmune diseases, which are characterized by elevated MMP activity.
In this regard, use of the nanocrystalline silver of the present
invention, when in therapeutically effective amounts, provides the
dual modulation of MMP-9 and TNF-.alpha. to alleviate the particular
condition.

Additional Examples

Example 1

6 milligrams of antimicrobial metals with atomic disorder, in free-
standing powder form, are sprinkled lightly onto 6.5 cm2 of burned
tissue, and thereafter wet with a light spray of water or wound
exudate or TDWL (Trans Dermal Water Loss) or other bodily fluids, so
as to provide an antimicrobial treatment to the burned tissue. The
treatment is repeated every 24 hours until the therapeutic effects are
no longer needed.

Example 2

0.5 milligrams of antimicrobial metals with atomic disorder, in free-
standing powder form, are injected, using a small-needle drug delivery
system or a needle-less drug delivery system, into gum tissue so as to
treat gingivitis. The treatment is repeated every 3 days until the
therapeutic effects are no longer needed.

Example 3

A solution of antimicrobial metals with atomic disorder is prepared by
dissolving 6 milligrams of antimicrobial metals with atomic disorder
in 1 gram of water. The solution of antimicrobial metals with atomic
disorder is applied as a rinse or bath or wash to a wound site so as
to provide an antimicrobial treatment to the wound site. The treatment
is repeated every 24 hours until the therapeutic effects are no longer
needed.

Example 4

A solution of antimicrobial metals with atomic disorder is prepared by
dissolving 6 milligrams of antimicrobial metals with atomic disorder
in 1 gram of water. The solution of antimicrobial metals with atomic
disorder is applied to the interior of the bladder via a catheter so
as to provide antimicrobial treatment to the bladder. The treatment is
repeated every 8 hours until the therapeutic effects are no longer
needed.

Example 5

A solution of antimicrobial metals with atomic disorder is prepared by
dissolving 6 milligrams of antimicrobial metals with atomic disorder
in 1 gram of water. The solution of antimicrobial metals with atomic
disorder is injected (using a small-needle or needle-less injection
system) under the toenails or into the nail bed and/or the surrounding
tissue of a person suffering from onychomycosis so as to provide an
antimicrobial treatment to the tissue. The treatment is repeated 2
times a day until the therapeutic effects are no longer needed.

Example 6

Summary

Solutions of nanocrystalline noble metals were prepared by immersing
Acticoat.RTM. burn dressings (distributed by Smith & Nephew) in
reverse osmosis water that had been pretreated with CO2 in order to
reduce the pH. Two different concentrations of antimicrobial metals
with atomic disorder solutions were prepared by this method, the
concentrations being 85 mg/mL and 318 mg/mL. Solutions of silver
nitrate were also prepared to use as comparisons in the experiments.
The concentrations of the silver nitrate were 103 ppm of silver and
295 ppm of silver as determined by Atomic Absorption Spectroscopy.

The solutions were in turn placed in an ultrasonic nebulizer that
created small droplets containing dissolved and suspended parts of the
solution of nanocrystalline noble metals. The output from the
nebulizer was directed into a chamber made from a stainless steel
frame and base. Petri dishes containing Mueller Hinton agar streaked
with 4 h old cultures of Pseudomonas aerugiona and Staphylococcus
aureus were exposed to nanocrystalline noble metal aerosols and the
silver nitrate aerosols.

The results of the tests show that nanocrystalline noble metal
aerosols transmit the antimicrobial activity of the dressings to
remote sites, and nanocrystalline noble metal aerosols are more
effective than comparable concentrations of silver nitrate.

Introduction

In many instances the delivery of antimicrobial materials may most
expeditiously be accomplished by using aerosols (e.g., in the
treatment of pneumonia). The drawback of aerosols is the requirement
for a high concentration of the active ingredient to ensure that a
sufficient amount is delivered to achieve the biological effect
desired without causing problems with the carrier solvent (e.g.,
water). The essential requirement of the equipment for producing an
aerosol that contains dissolved and suspended components of
antimicrobial metals with atomic disorder is that it must form
droplets of aerosol directly from the liquid form, and the aerosol
droplets must be small enough to reach the lungs. This means that the
droplets should be preferably less than approximately 10 mm. To meet
these requirements, the aerosol cannot be created by first evaporating
the liquid and then condensing it to form droplets, since this would
remove the desired antimicrobial metals with atomic disorder from the
aerosol. There are two methods that can be used to relatively easily
form the droplets directly: (1) mechanical disruption of the liquid;
and (2) air, under pressure, passing through some form of orifice that
combines the air and the liquid in a way that creates droplets instead
of evaporating the liquid.

Several experiments were carried out with antimicrobial metals with
atomic disorder and silver nitrate solutions to determine if the
antimicrobial activity of the dressing could be transferred through a
direct droplet aerosol to a Petri dish.

Equipment

The method used to create an aerosol for these tests was the
mechanical method in the form of an ultrasonic nebulizer. For
convenience, an ultrasonic humidifier was used. The liquid containing
the dissolved and suspended antimicrobial metals with atomic disorder
was placed in the water reservoir of the humidifier. When power was
applied to the humidifier, aerosol droplets of dissolved and suspended
antimicrobial metals with atomic disorder were generated and flowed
from the output nozzle.

A test chamber was constructed using a stainless steel frame with a
transparent plastic covering. The frame was placed on a stainless
steel plate. The output nozzle from the humidifier was modified so
that the aerosol could be directed into the chamber at a height of
approximately 30 cm from the base. The plates and other test samples
were is placed on the stainless steel plate and exposed to the aerosol
for a prescribed length of time. Solution 1

A solution of antimicrobial metals with atomic disorder was prepared
by immersing 518 sq. inches of Acticoat.RTM. burn dressing in 1 L of
reverse osmosis water, which was treated with CO2 to maintain a pH of
6.5. After 20 minutes the concentration of silver in the water was 85
mg/mL.

Solution 2

A solution containing 370 mg/mL of silver from a Acticoat.RTM.
dressing was prepared as follows: 1 L of reverse osmosis water was
purged with commercial grade carbon dioxide until the pH was 4.3.
Sufficient Acticoat.RTM. dressing was added to bring the pH up to 6.5.
At that time, the silver concentration was 370 mg/mL.

Solution 3

Ag as AgNO.sub.3 was prepared by dissolving 0.157 g of AgNO.sub.3 into
1 L of reverse osmosis water and mixed until dissolved. The solution
was analyzed by Atomic Absorption Spectroscopy and found to be 102.9
ppm of silver.

Solution 4

Ag as AgNO.sub.3 was prepared by dissolving 0.427 of AgNO.sub.3 into 1
L of reverse osmosis water and mixed until dissolved. The solution was
analyzed by Atomic Absorption Spectroscopy and found to be 295 ppm of
silver.

Aerosolization

Petri dishes, containing Mueller Hinton agar, were streaked with 4 h
old cultures of Pseudomonas aeruginosa or Staphylococcus aureus. The
plates were then weighed and their exposed outer surfaces were coated
with Parafilm to prevent condensation from occurring on these
surfaces. These plates were placed in the aerosol chamber uncovered.
The ultrasonic nebulizer was then started and run for 53 minutes. The
plates were then removed from the chamber, the plastic was removed and
the dishes re-weighed so that the amount of moisture loss/gain could
be determined.

The plates were then placed in a 35.degree. C. incubator for 16 h.
After incubation the pattern and amount of growth was assessed on the
plates for both organisms.

Viability Assessment

Three of the six plates made for each organism were tested to
determine if the antimicrobial effect was cidal or static in nature.
This was accomplished by rinsing or placing a piece of the clear
section of agar in the Petri dish plates into Tryptic soy broth in a
test tube and incubating for 4 h or 16 h. If the medium turned turbid
in 4 h it would indicate that the antimicrobial affect was
bacteriostatic in nature. If the organism took more than 16 h to grow,
as indicated by turbidity, it was considered an indication that both
static and cidal effects occurred. If no growth occurred, the effect
was bactericidal.

Results

The results for Solutions 1 and 3 are summarized in the following two
table.

TABLE-US-00017 Antimicrobial Metals With Atomic Disorder AgNo.sub.3
Ps. S. Ps. S. Organism Aeruginosa aureus Aeruginosa aureus Ag
concentration 85 85 99 99 (.mu.g/mL) pH of test solution 6.5 6.5
Approx. Approx. 6.5 6.5 Exposure time 53 53 53 53 (minutes) Exposed
area (sq. in) 9.8 9.8 9.8 9.8 Weight gain (g) 0.8 0.8 1.05 1.05 Growth
at 16 h 0 0 0 ++++ (0-++++) at 48 h 0 ++ 0 ++++ Viable 4 h No Yes No
Yes 16 h Yes Yes Yes Yes

The results for Solutions 2 and 4 are summarized in the following two
table.

TABLE-US-00018 Antimicrobial Metals With Atomic Disorder AgNo.sub.3
Ps. S. Ps. S. Organism aeruginosa aureus aeruginosa aureus Ag
concentration 370 370 300 300 (.mu.g/mL) pH of test solution 6.5 6.5
Approx. Approx. 6.3 6.3 Exposure time 53 53 53 53 (minutes) Exposed
area (sq in) 9.8 9.8 9.8 9.8 Weight gain (g) 1.14 1.14 1.12 1.12
Growth at 16 h 0 0 0 0 (0-++++) at 48 h 0 0 0 +++ Viable 4 h No No No
No 16 h No No No N/A

Discussion

At the low concentration of silver in solution, the Acticoat.RTM.
dressing generated silver was effective at controlling the growth of
both organisms while the silver nitrate only prevented the growth of
Ps. aeruginosa. Viability tests showed that at the low concentration,
neither form of silver was completely bactericidal although the poor
growth on the plates treated with antimicrobial metals with atomic
disorder compared to the silver nitrate treated plates suggests that a
significant log reduction occurred in the plates treated with the
aerosol of antimicrobial metals with atomic disorder.

At a higher concentration of silver, both antimicrobial metals with
atomic disorder (370 mg/mL) and AgNO.sub.3 (300 mg/mL) were effective
at controlling P. aeruginosa. Since no re-growth occurred, it is
assumed that the agent at this concentration was bactericidal.
Antimicrobial silver with atomic disorder was more effective than
AgNO.sub.3 at controlling S. aureus. No re-growth occurred on any
plates or in the broth indicating a total kill of the organism while,
in the AgNO.sub.3 treatment, a large number of organisms grew at 16 h.

Based on weight gain during aerosol treatments, a dose per unit area
can be calculated. In each case for Solution 1, the dose was 8.5 mg/
sq. inch, while for Solution 2, the dose was 38 mg/sq. inch. These
doses, on a per lung basis, would be less than 10 mg of silver per
hour of treatment. Each hour of treatment with antimicrobial silver
with atomic disorder aerosols appears to provide at least 48 h of
protection. Therefore, the dose per day, from the high concentration
treatment, would be about 5 mg or a little less than the silver
released by 2 sq. inches of SSD per day.

The most significant advantage of using antimicrobial silver with
atomic disorder may be the lack of a toxic action such as NO.sub.3 or
sulfadiazine.

Conclusions

(1) Aerosols of antimicrobial metals with atomic disorder transmit the
antimicrobial activity of the dressings to remote sites.

(2) Aerosols of antimicrobial metals with atomic disorder are more
effective than comparable concentrations of silver nitrate.

(3) The dose delivered is acceptable and would not appear to be
excessive.

(4) No toxic cations (NO.sub.3 or sulfadiazine) are introduced to the
patient.

Example 7

(Gels of Antimicrobial Metals with Atomic Disorder)

Gel products of antimicrobial metals with atomic disorder encompass
both wet and dry materials.

A wet gel product of antimicrobial metals with atomic disorder is a
product that provides moisture to a dry skin condition (psoriasis,
eczema, acne, wound, etc.) and facilitates autolytic debridement of
necrotic tissue. It also delivers the antimicrobial and anti-
inflammatory properties of the suspended antimicrobial metals with
atomic disorder powders.

In many instances it is also beneficial to supply biologically active
molecules to elicit a specific response such as cell migration, etc.
Since these biologically active molecules are susceptible to microbial
degradation if not protected, it is beneficial to include them in gels
of antimicrobial metals with atomic disorder that will provide the
necessary protection.

Dry gel products of antimicrobial metals with atomic disorder are
physically stabilized (dry or cross-linked) materials that provide
lubricious, antimicrobial, antithrombogenic, and anti-inflammatory
properties to a variety of implantable, trancutaneous or topically
applied devices. The coatings may also provide other benefits such as
accelerating or otherwise facilitating tissue integration by creating
a favorable environment for cell proliferation. This favorable
environment may be created by including cyto-conductive agents or anti-
adhesion agents such as bone morphogenetic proteins, B-glucan
hyaluronic acids in the gel. The gel may be stabilized by cross-
linking the gel components (collagen, gelatin, etc.) or by drying the
coated materials.

Examples of the primary gelling agents are listed in the following
table. Biologically active ingredients that may be used, in any
combination with the primary gelling agents, are given in the
subsequent table. Materials that should not be used with gels of
antimicrobial silver with atomic disorder are given in the final
table.

TABLE-US-00019 Percentage Composition Material Carboxymethyl cellulose
(CMC) 0.1-10 Polyvinyl alcohol (PVA) 0.1-10 Collagen 0.1-10 Pectin
0.1-10 Gelatin 0.1-10 Chitin 0.1-10 Chitosan 0.1-10 Alginate 0.1-10
Poly (.alpha.-amino acids) Polyester Poly-1-caprolactone PEG Cocoa
butter Sepigel Biologically Active Ingredients Methyl paraben <3
Propyl paraben <3 B-glucan <5 Hyaluronic acid <5 Epidermal growth
factor <1 Platelet derived growth factor <1 Transforming growth factor
<1 Vascular endothelial growth factor <1 Interleukins <1 Heparin <5
Bone morphogenetic proteins <1 Non-Compatible Materials Chloride salts
>0.01 Aldehydes >0.01 Ketones >0.01 Long chain alcohols >0.01 Glycerol
>0.01 Triethanolamine >0.01

Example 8

(Examples of Gels with Antimicrobial Metals with Atomic Disorder)

Gels were prepared as described above in Example 11 in the Treatment
of Inflammatory Skin Conditions examples above.

Other embodiments are in the claims.



United States Patent 6,669,966
Antelman December 30, 2003
Compositions for facilitating skin growth and methods and articles
using same

Abstract

Skin-growth-enhancing compounds and compositions including a
therapeutically effective amount of at least one electron active
compound, or a pharmaceutically acceptable derivative thereof, that
has at least two polyvalent cations, at least one of which has a first
valence state and at least one of which has a second, different
valence state. Preferred compounds include Bi(III,V) oxide, Co(II,III)
oxide, Cu(I,III) oxide, Fe(II,III) oxide, Mn(II,III) oxide, and
Pr(III,IV) oxide, and Ag(I,III) oxide, or a combination thereof. These
compounds may be in a crystalline state having metallic cations of two
different valences, or electronic states, in the inorganic crystal.
Also included are articles containing such compositions, such as wound
dressings, and methods for facilitating or enhancing skin growth using
these compounds, compositions, and articles, such as for the treatment
or management of burns or skin grafts.
Inventors: Antelman; Marvin S. (Rehovot, IL)
Assignee: Marantech Holding LLC (East Providence, RI)
Appl. No.: 09/692,127
Filed: October 20, 2000
Related U.S. Patent Documents

Application Number Filing Date Patent Number Issue Date
552172 Apr., 2000 6258385

Current U.S. Class: 424/635 ; 424/405; 424/407; 424/443; 424/445;
424/446; 424/447; 424/489; 424/600; 424/613; 424/617; 424/639;
424/646; 424/647; 424/648; 424/653; 424/724; 424/725; 424/DIG.13; 424/
DIG.6; 514/165; 514/772; 514/772.3; 514/788.1; 514/836; 514/904;
514/905; 514/925; 514/928; 514/950
Current International Class: A61K 47/02 (20060101); A61K 8/02
(20060101); A61K 8/19 (20060101); A61Q 1/12 (20060101); A61K 33/24
(20060101); A61K 33/26 (20060101); A61K 33/32 (20060101); A61K 33/34
(20060101); A61K 33/38 (20060101); A61K 9/06 (20060101); A61Q 19/00
(20060101); A61K 033/34 (); A61K 009/00 (); A61K 031/60 (); A61K
033/00 (); A61K 035/78 (); A61K 047/00 (); A61L 015/00 (); A01N 025/00
()
Field of Search:
424/600,617,618,630,635,639,646,647,648,653,405,407,443,445,446,447,489,613
514/165,772,772.3,788.1,836,904,905,925,928,950
References Cited [Referenced By]
U.S. Patent Documents

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4447254 May 1984 Hughes et al.
4828832 May 1989 De Cuellar et al.
4952411 August 1990 Fox, Jr. et al.
5017295 May 1991 Antelman
5073382 December 1991 Antelman
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5098582 March 1992 Antelman
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759, and 760. .
Gennaro, A., Remington's Pharmaceutical Sciences, Easton, PA: Mack
Publishing Company, 1985, 17.sup.th Edition, p. 1573-1575, 1585-1594,
and 1601. .
Antelman, Marvin S.; "Silver (II,III) Disinfectants"; Soap/Cosmetics/
Chemical Specialties, Mar. 1994, pp. 52-59. .
Antelman, Marvin S.; Abstracts of the American Chemical Society;
1992(203). .
Antelman, Marvin S.; "Anti-Pathogenic Multivalent Silver Molecular
Semiconductors"; Precious Metals; 1992(16); pp. 141-149. .
Antelman, Marvin S.; "Multivalent Silver Bacterides"; Precious Metals;
1992(16); pp. 151-163. .
Fung, Man C. and Bowen, Debra L.; "Silver Products for Medical
Indications: Risk-Benefit Assessment", Clinical Toxicology, 1996, pp.
119-126..

Primary Examiner: Clardy; S. Mark
Assistant Examiner: Choi; Frank
Attorney, Agent or Firm: Pennie & Edmonds LLP
Parent Case Text


CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation-in-part of application Ser. No.
09/552,172, filed Apr. 18, 2000, now U.S. Pat. No. 6,258,385, and
claims the benefit of Provisional Application Nos. 60/174,793, filed
Jan. 6, 2000, 60/184,053, filed Feb. 22, 2000, and 60/214,503, filed
Jun. 28, 2000.
Claims


What is claimed is:

1. A method for facilitating or enhancing growth of a patient's skin,
which comprises administering at least one electron active compound
that has at least two polyvalent cations, at least one of which has a
first valence state and at least one of which has a second different
valence state, said at least one electron active compound being
administered in an amount and for a period of time which is
therapeutically effective to facilitate or enhance skin growth, and
wherein the electron active compound comprises at least one of
Bi(III,V) oxide, Co(II,III) oxide, Cu(I,III) oxide, Fe(II,III) oxide,
Mn(II,III) oxide, or Pr(III,IV) oxide.

2. The method of claim 1, wherein the patient is a mammal and wherein
the therapeutically effective amount of the electron active
compound(s) administered is from about 1 ppm to 500,000 ppm.

3. The method of claim 2, wherein the therapeutically effective amount
is from about 50 ppm to 100,000 ppm.

4. The method of claim 2, wherein the mammal is a human and wherein
the at least one electron active compound is administered topically or
transdermally.

5. The method of claim 1, wherein the method further comprises
administering at least one additional different therapeutic agent
present in an amount sufficient to facilitate or enhance growth of the
patient's skin and wherein the at least one additional therapeutic
agent comprises at least one of a chelating agent, a vitamin, a
mineral, silica hydride microclusters, an analgesic, elderberry
extract, and aspirin.

6. The method of claim 5, wherein the at least one additional
therapeutic agent is administered concurrently with the at least one
electron active metal oxide compound.

7. The method of claim 1, further which comprises combining the at
least one electron active compound with a carrier medium before
administration to the patient, wherein the carrier medium comprises
petroleum jelly or a thixotropic agent sufficient to increase
adherence of the composition to the skin without excessive runoff.

8. The method of claim 1, wherein the composition is administered
topically directly to the skin in the form of a powder.

9. The method of claim 1, wherein the administering comprises
application of the at least one electron active compound to the skin
at a dosage level of about 10 mg to 500 mg per cm.sub.2 of skin
surface.

10. The method of claim 1, wherein the amount is insufficient to cause
adverse effects.

11. The method of claim 1, wherein the facilitating or enhancing of
skin growth comprises the treatment or management of a burn or skin
graft, or a symptom thereof.

12. A skin-growth-enhancing composition comprising: (a) a
therapeutically effective amount of at least one electron active
compound that has at least two polyvalent cations, at least one of
which has a first valence state and at least one of which has a second
different valence state, the at least one electron active compound
being present in an amount which is therapeutically effective to
facilitate or enhance skin growth, and wherein the electron active
compound comprises at least one of Bi(III,V) oxide, Co(I,III) oxide,
Cu(II,III) oxide, or Pr(III,IV) oxide; and (b) a carrier comprising at
least one of a petroleum jelly or a thixotropic agent present in an
amount sufficient to increase adherence of the composition to skin
without excessive runoff.

13. The skin-growth-enhancing composition of claim 12, wherein the at
least one electron active compound is present in an amount of from
about 1 ppm to 500,000 ppm.

14. The skin-growth-enhancing composition of claim 12, in the form of
a powder, or a plurality of powder crystals or granules.

15. The skin-growth-enhancing composition of claim 12, further
comprising an oxidizing agent present in an amount sufficient to
enhance efficacy of the active compound but insufficient to cause skin
irritation.

16. A wound dressing comprising the composition of claim 12.

17. The wound dressing of claim 16, wherein the wound dressing
comprises an adhesive-containing bandage, a cotton roll bandage, or a
gellable polymer.

18. A dressing comprising at least one electron active compound, the
electron active compound having at least two polyvalent cations, at
least one of which has a first valence state and at least one of which
has a second different valence state, wherein the electron active
compound comprises at least one of Bi(III,V) oxide, Co(II,III) oxide,
Cu(I,III) oxide, Mn(II,III) oxide, or Pr(III,IV) oxide.

19. The dressing of claim 18, wherein the dressing comprises a
therapeutically sufficient amount of the electron active compound to
manage, or treat at least one of wound, a burn, a lesion, an ulcer, a
sore, a boil, a wart, or combination thereof.

20. The dressing according to claim 18, further comprising an
oxidizing agent present in an amount sufficient to enhance efficacy of
the electron active compound but insufficient to cause skin
irritation.

21. The dressing according to claim 18, wherein the dressing comprises
at least one of a bandage or a gellable polymer.

22. The dressing according to claim 18, wherein the electron active
compound is in the form of a powder, a plurality of powder crystals,
granules, or combination thereof.

23. A method treating or managing one or more conditions, which
comprises topically administering to a patient at least one dressing
comprising at least one electron active compound, the electron active
compound having at least two polyvalent cations, at least one of which
has a first valence state and at least one of which has a second
different valence state, wherein the electron active compound
comprises at least one of Bi(III,V) oxide, Co(II,III) oxide, Cu(I,III)
oxide, Mn(II,III) oxide, or Pr(III,IV) oxide, and wherein the
condition comprises at least one of a wound, a burn, a lesion, an
ulcer, a sore, a boil, a wart, or combination thereof.

24. The method according to claim 23, wherein each electron active
compound is present in a therapeutically effective amount.

25. The method of claim 23, wherein the dressing comprises at least
one of a bandage or a gellable polymer.
Description


FIELD OF THE INVENTION

The invention relates to skin-growth-enhancing compositions including
certain compounds having polyvalent cations in their crystal lattices,
particularly certain inorganic metal oxides. Articles and methods of
facilitating or enhancing skin growth to treat or manage certain
conditions, such as burn therapy or skin grafts, using such skin
growth compositions are included in the invention.

BACKGROUND OF THE INVENTION

Animal and mammalian skin, in particular, human skin, is a
multifunctional organ. Not only does the skin provide an external
covering to protect the body, but it also performs several specialized
functions, such as breathing, perspiring, sensory information
processing, and oil production. Oil production, essential to the
protective features of the skin, works when an oily substance known as
seburn is released from the sebaceous glands, which are large glands
located at the base of a hair follicle. This permits the skin to
moisturize and waterproof itself, thereby protecting itself from the
environment.

The skin is the most environmentally-stressed organ in mammals,
particularly in humans. The skin is subjected to toxic chemicals and
hostile environments, as well as being the only organ directly exposed
to Ultraviolet ("UV") light in the presence of oxygen. Lengthy
exposure of the skin to UV light typically damages the skin,
resulting, in sunburn, photoaging, carcinogenesis, and other related
skin disorders.

In particular, human skin is a composite material of the epidermis and
the dermis. The topmost part of the epidermis is the stratum corneum.
This layer is the stiffest layer of the skin, as well as the one most
affected by the surrounding environment. Below the stratum corneum is
the internal portion of the epidermis. Below the epidermis, the
topmost layer of the dermis is the papillary dermis, which is made of
relatively loose connective tissues that define the micro-relief of
the skin. The reticular dermis, disposed beneath the papillary dermis,
is tight, connective tissue that is spatially organized. The reticular
dermis is also associated with coarse wrinkles. At the bottom of the
dermis lies the subcutaneous layer.

The principal functions of the skin include protection, excretion,
secretion, absorption, thermoregulation, pigmentogenesis,
accumulation, sensory perception, and regulation of immunological
processes. These functions are detrimentally affected by the
structural changes in the skin due to aging and excessive sun
exposure. The physiological changes associated with skin aging include
impairment of the barrier function and decreased turnover of epidermal
cells, for example.

The mechanical properties of the skin, such as elasticity, are
believed to be controlled by the density and geometry of the network
of collagen and elastic fiber tissue therein. Damaged collagen and
elastin lose their contractile properties, resulting in skin wrinkling
and skin surface roughness. As the skin ages or becomes unhealthy, it
acquires sags, stretch marks, burns, bruises or wrinkles, it roughens,
and it has reduced ability to synthesize Vitamin D. Aged skin also
becomes thinner and has a flattened dermoepidermal interface because
of the alterations in collagen, elastin, and glycosaminoglycans.

UV light exposure in the presence of oxygen results in the undesirable
creation of free radicals, which is believed to lead to various skin
disorders, diseases, or conditions. In the skin, these free radicals
frequently trigger the release of inflammatory mediators, commonly
manifested as sun burn; cytoskeletal alterations, breaking down the
collagen in the skin; and may also result in structural DNA changes,
such as DNA strand breaks and dimer formation. The body attempts to
neutralize the free radicals generated by UV light through the use of
antioxidants. Antioxidants are commonly found in two forms--enzymatic
and non-enzymatic.

Various ingredients have been used alone or in certain combinations to
form pharmaceuticals designed to prevent and treat certain cellular,
skin, and other conditions, such as burns. Although a variety of
compositions and methods for treating various skin conditions are
presently available to those of ordinary skill in the art, the
treatments are often not completely effective and often involve
adverse effects, such as overdrying of the skin. Furthermore, some
existing treatments simply address the symptoms and fail to treat the
underlying condition, as well as helping to reduce the incidence of
remission or the appearance of recurring or new disorders.

Multivalent silver molecules have also been disclosed for various
uses, as they are reported to be non-toxic to animals and humans. M.
Antelman, "Anti-Pathogenic Multivalent Silver Molecular
Semiconductors," Precious Metals, vol. 16:141-149 (1992); M. Antelman,
"Multivalent Silver Bactericides," Precious Metals, vol. 16:151-163
(1992). For example, tetrasilver tetroxide activated with an oxidizing
agent is disclosed for use in bactericidal, fungicidal, and algicidal
use, such as in municipal and industrial water treatment applications
and for the treatment of AIDS.

A variety of sources also report the use of certain divalent silver
compounds for water treatment, as well as the use of such compounds,
typically in combination with certain oxidizing agents, metals, or
other compounds, as disinfectants, bactericides, algicides, and
fungicides. One source also reports a single in vitro study of the use
of such compounds for the treatment of AIDS. These sources include M.
Antelman, "Silver (II, III) Disinfectants," Soap/Cosmetics/Chemical
Specialties, pp. 52-59 (Mar., 1994), and U.S. Pat. Nos. 5,017,295;
5,073,382; 5,078,902; 5,089,275; 5,098,582; 5,211,855; 5,223,149;
5,336,416; and 5,772,896.

U.S. Pat. No. 5,336,499 discloses tetrasilver tetroxide and persulfate
compositions having certain in vitro anti-pathogenic properties, i.e.,
bactericidal, fungicidal, viricidal, and algicidal, in certain
concentrations as low as 0.3 ppm, particularly in nutrient broth
cultures. The persulfate or another oxidizing agent is required to
activate the tetroxide crystals. Also disclosed are: an in vitro study
regarding the inhibition of yeast growth in nutrient broth and the
formulation of a gynecological cream and douche based on these
results, and a report of an in vitro AIDS test with the compositions
indicating total suppression of the virus at 18.0 ppm.

U.S. Pat. No. 5,571,520 discloses the use of molecular crystals of
tetrasilver tetroxide, particularly with oxidizing agents to enhance
the efficiency of such devices, for killing pathogenic microorganisms,
such as staph infections. Amounts of 10 ppm sodium persulfate as an
oxidizing agent were used with certain amounts of silver tetroxide in
the reported in vitro testing. One human study involved in vivo curing
of a gynecological yeast infection with 10 ppm of the silver tetroxide
and 40 ppm sodium persulfate. Other in vivo topical studies report in
conclusory fashion the cure of a single case of athlete's foot with a
solution of 100 ppm of the composition and the cure of a single case
of toenail fungus with a 25% suspension of the composition.

U.S. Pat. No. 5,676,977 discloses intravenously injected tetrasilver
tetroxide crystals used for destroying the AIDS virus, AIDS
synergistic pathogens, and immunity suppressing moieties (ISM) in
humans. The crystals were formulated for a single injection at about
40 ppm of human blood. This reference also discloses the compositions
cause hepatomegaly, also known as enlarged liver, albeit with no
reported loss of liver function.

The aforementioned references report detailed descriptions of the
mechanism via which the multivalent silver molecular crystal devices
were believed to operate. A discussion of such results and concepts
was presented at a Seminar entitled "Incurable Diseases
Update" (Weizmann Institute of Science, Rehovot, Israel, Feb. 11,
1998). The title of this presentation was "Beyond Antibiotics, Non
Toxic Disinfectants and Tetrasil.TM. (a composition including
tetrasilver tetroxide)." In this article, it was reported that the
effects of the electron transfer involved with respect to the
tetroxide, rendered it a more powerful germicide than other silver
entities. Other patents cover multivalent silver antimicrobial
compositions, e.g., U.S. Pat. Nos. 5,017,295 for Ag(II) and 5,223,149
for Ag (III). These are stronger antimicrobial agents than Ag (I)
compounds, but they pale by comparison to tetrasilver tetroxide.

Likewise, colloidal silver that derives its germicidal properties from
trace silver (I) ions it generates in various environments is also
less effective. Accordingly, the oligodynamic properties of these
entities may be summarized as follows, which is referred to as the
Horsfal series:

Another property of the tetrasilver tetroxide is that it does not
stain organic matter such as skin in like manner as Ag(I) compounds
do. In addition, it is light stable.

Further, synthetic routes for making Bi(III,V) oxide are detailed and
reviewed in Gmelins Handbuch DerAnorganischen Chemie, vol. 16:642
(1964). Also, Co(II,III) oxide, Fe(II,III) oxide, Mn(II,III) oxide,
and Pr(III,IV) oxide can all be found in nature. These five
multivalent metal oxides are also all available commercially.

Certain skin conditions, such as burns, skin cancer, and skin grafts,
however, require the growth or regrowth of damaged or eradicated
tissue. Burns to skin are caused by thermal, chemical, or electrical
contact, which results in, for example, protein denaturation, burn
wound edema, loss of intravascular fluid volume due to increase
vascular permeability, and combinations thereof. Systemic effects, for
example, hypovolemic shock, infection, respiratory tract injury, or a
combination thereof, pose a greater threat to the life of the victim
that do the above-noted local effects.

In spontaneous burn wound healing, dead tissue sloughs off as new
epithelium begins to cover the injured area. In superficial burns,
regeneration or growth of skin tissue occurs rapidly from, for
example, uninjured epidermal elements, hair follicles, and sweat
glands. Minimal scarring typically results unless infection occurs
during the healing process. With deep burns, i.e., destruction of the
epidermis and much of the dermis, reepithelialization typically begins
from the edges of the wound or from the scattered remains of
integument. The process is typically slow, and excessive granulation
tissue often forms before being covered by new epithelium. Such wounds
generally contract and develop into disfiguring or disabling scars
unless treated promptly by, for example, skin grafting. Unfortunately,
some skin grafts are rejected by the host's body in the absence of
immune suppression treatment, which adds additional expense to the
treatment and often creates additional adverse effects in the patient.

The severity of a burn is judged by the quantity of tissue involved.
This quantity is represented by the percentage of body surface area (%
BSA) burned and by the depth of the burn. A conventional
classification of burns by severity is: small burn, or less than 15%
BSA; moderate burn, or 15% to 49% BSA; large burn, or 50% to 69% BSA;
and massive burn, or greater than 70% BSA.

The depth of a burn may be described as a first, second, or third
degree burn. First degree burns are red, very sensitive to the touch,
and usually moist. Blisters typically do not form and the surface
markedly and widely blanches under light pressure. Second degree burns
may or may not have blisters, but the wound base is sensitive to touch
and may blanch to pressure. Third degree burns may, but generally do
not, present blisters. The skin surface may be white and pliable when
pressure is applied, or it may be black, charred, and leathery. Third
degree burns may be pale in color and even mistaken for normal skin,
but the subdermal vessels do not blanch to pressure. The wound may
alternatively be bright red, due to fixed hemoglobin in the subdermal
region. The third degree burns are generally anasthetic or
hypoesthetic, with hair being easily pulled from the follicles. Often,
the distinction between deep second and third degree burns can be made
only after 3 to 5 days of observation.

U.S. Pat. No. 4,828,832 to De Cuellar et al. discloses metallic silver
particles and an oxidizing agent, such as benzoyl peroxide, dispersed
in a carrier for application to a skin lesion, such as for the
treatment of burns.

The above-noted compositions are not believed to have suitable
efficacy in treating or managing conditions that require skin growth,
such as the treatment or management of burns.

Thus, it is desired to find skin-growth-enhancing pharmaceutical
compositions and methods for facilitating or enhancing skin growth to
treat or manage one or more dermatological conditions. It is also
desired to facilitate or enhance the rate of skin growth while
avoiding adverse effects present when administering certain
conventional treatments or skin replacement.

SUMMARY OF THE INVENTION

The invention relates to methods for facilitating or enhancing skin
growth of a patient's skin, by administering at least one electron
active compound, or a pharmaceutically acceptable derivative thereof,
that has at least two polyvalent cations, at least one of which has a
first valence state and at least one of which has a second different
valence state, to treat or manage the condition, or a symptom thereof,
in an amount and for a period of time which is therapeutically
effective to facilitate or enhance skin growth.

In one embodiment, the patient is a mammal and the therapeutically
effective amount of the electron active metal oxide compound(s)
administered is from about 1 ppm to 500,000 ppm. In another
embodiment, the therapeutically effective amount is from about 50 ppm
to 100,000 ppm. In yet another embodiment, the mammal is a human and
the at least one electron active metal oxide compound is administered
topically, parenterally, or transdermally.

In one embodiment, the method further includes administering at least
one additional different therapeutic agent present in an amount
sufficient to facilitate or enhance the treatment or management of the
condition. It is possible to administer the at least one additional
therapeutic agent concurrently with the at least one electron active
metal oxide compound, although in other embodiments administration may
be sequential in either order.

Preferably, the at least one electron active compound includes a metal
oxide. In one embodiment, the electron active compound includes at
least one of Bi(III,V) oxide, Co(II,III) oxide, Cu(I,III) oxide,
Fe(II,III) oxide, Mn(II,III) oxide, or Pr(III,IV) oxide, or a
pharmaceutically acceptable derivative thereof.

It is also possible to combine the at least one electron active
compound with a carrier medium before administration to the patient.
In one preferred embodiment, the carrier medium includes petroleum
jelly. In some embodiments, no carrier is required, and the
composition is administered in the form of a powder, or a plurality of
powder crystals or granules. In one topical embodiment, the carrier
medium includes a thixotropic agent sufficient to increase adherence
of the composition to the skin without excessive runoff. The at least
one electron active compound may be applied to the skin at a dosage
level of about 10 mg to 500 mg per cm.sup.2 of skin surface.
Preferably, the therapeutically effective amount administered is
insufficient to cause adverse effects.

Preferably, the facilitating or enhancing of skin growth comprises the
treatment or management of a burn or skin graft. In one embodiment, a
pathogen is killed concurrently with the treatment or management of a
burn or skin graft. Alternatively, the growth of a pathogen is halted,
diminished, or inhibited concurrently with the treatment or management
of a burn or skin graft.

The present invention also relates to skin-growth-enhancing
compositions that include at least one electron active compound, or a
pharmaceutically acceptable derivative thereof, that has at least two
polyvalent cations, at least one of which has a first valence state
and at least one of which has a second, different valence state, in an
amount and for a period of time which is therapeutically effective to
facilitate or enhance skin growth. Advantageously, the pharmaceutical
composition may have antipathogenic efficacy. Preferably, the at least
one electron active compound includes a metal oxide. In one
embodiment, the metal oxide includes at least one of bismuth, cobalt,
copper, iron, manganese, praseodymium, or a combination thereof.
Preferably, in that embodiment, the metal oxide includes at least one
of Bi(III,V) oxide, Co(II,III) oxide, Cu(I,III) oxide, Fe(II,III)
oxide, Mn(II,III) oxide, Pr(III,IV) oxide, Ag(I,III) oxide, or a
combination thereof. Alternately, the pharmaceutical composition does
not include tetrasilver tetroxide. In another alternate embodiment,
the pharmaceutical composition does not include tricobalt tetroxide.
In one embodiment, the pharmaceutical composition may include at least
two different electron active compounds. In another embodiment, the
compound may be in powder, powder crystal, or granular form.

In a preferred embodiment, the first valence and the second valence of
the at least two polyvalent cations differ by at least 1, preferably
by 1 or 2. In another preferred embodiment, the first valence and the
second valence of the at least two polyvalent cations differ by more
than 2. Advantageously, the electron active compound has at least one
polyvalent cation which has an EMF.sup.OX of at least about +0.1
Volts.

In one embodiment, the amount of the at least one electron active
compound is present in an amount from about 1 ppm to 500,000 ppm,
based on the weight of the composition. If desired, the pharmaceutical
composition can include a pharmaceutically acceptable carrier. In one
embodiment, the carrier includes a petroleum jelly. In one preferred
embodiment, the carrier medium is adapted for topical administration
comprises a thixotropic agent sufficient to increase adherence of the
composition to skin without excessive runoff. Optionally, the
composition can also include an oxidizing agent, preferably present in
an amount sufficient to enhance the efficacy of the active compound
but insufficient to cause skin irritation. Preferably, the oxidizing
agent includes a peroxy acid salt of a persulfate.

The invention also relates to articles including the skin-growth-
enhancing compounds or compositions according to the invention. One
preferred embodiment includes a wound dressing including the at least
one compound or composition of the invention. In a more preferred
embodiment, the wound dressing includes an adhesive-containing
bandage, a cotton roll bandage, or a gellable polymer. The gellable
polymer may be any polymer, or combination thereof, available to those
of ordinary skill in the art, that has sufficiently low viscosity to
flow onto a skin area requiring treatment or management and that
subsequently thickens sufficiently upon application to a wound so as
to remain substantially affixed to the wound for a time sufficient to
provide treatment or management to the skin condition. The thickening
may occur, for example, by exposure to air, moisture in the air or
wound, by combination of two polymers directly on the afflicted skin
area, or due to heat from the afflicted skin area.

DEFINITIONS

Some of the terms used in connection with the invention can be defined
as follows:

The term "condition," as used herein, should be understood to refer to
a traditionally identified disease, as well as a disorder, an
affliction, or an ailment, particularly including those noted herein.
In particular, as used herein the term "condition" includes burns,
wounds, or sores, or a symptom thereof, such as where treatment or
management thereof requires skin growth. In one embodiment, condition
refers to burns or wounds other than mere sores.

The terms "prevent," "preventing," and "prevention," as used herein,
refer to stopping or hindering a condition, symptom, or pathogen
causing a condition, in a patient who is at risk of suffering from
such a condition. This also includes reducing the frequency or
severity, or both, of the occurrence of such conditions or one or more
symptoms thereof.

The terms "manage," "managing," and "management," as used herein,
includes controlling those conditions which cannot be cured
completely, reducing the time of affliction of such conditions, and
the like. Preferably, the compositions prevent, treat, or manage such
conditions without superficially discoloring the skin, i.e., no
discoloration to the naked eye. In one embodiment, the invention
relates to the treatment or management, while in another embodiment
the invention relates to the prevention, of the diseases or conditions
disclosed and claimed herein. The terms also include the use of the
compounds or compositions of the invention to facilitate the halting,
diminishing, or inhibiting of the growth or proliferation of pathogens
that may accentuate, amplify, exacerbate, or cause, either directly or
indirectly, a condition and/or a symptom thereof.

The term "patient" as used herein refers to animals, particularly to
mammals. In one preferred embodiment, the term patient refers to
humans.

The terms "adverse effects," "adverse side effects," and "side
effects," as used herein, include, but are not limited to, cardiac
arrhythmia, cardiac conduction disturbances, appetite stimulation,
weight gain, sedation, gastrointestinal distress, headache, dry mouth,
constipation, diarrhea, drug-drug interactions, superficial
discoloration of the skin, dry skin, hepatomegaly, fever, fatigue, and
the like. The term "cardiac arrhythmia" includes, but is not limited
to, ventricular tachyrhythmia, torsades de pointes, Q.sub.T
prolongation, and ventricular fibrillation.

The phrase "therapeutically effective amount" when used herein in
connection with the compositions and methods of the invention, means
that amount of electron active metal oxide compound(s) or
composition(s), or a derivative thereof, which, alone or in
combination with other drugs, provides a therapeutic benefit in the
treatment or management of a condition. In one embodiment, the
effective amount is one or more metal oxide compounds or compositions
as the sole active ingredient. Different therapeutically effective
amounts may be applicable for each condition, as will be readily known
or determined by those of ordinary skill in the art.

The term "substantially free" means less than about 10 weight percent,
preferably less than about 5 weight percent, more preferably less than
about 1 weight percent, and most preferably less than about 0.1 weight
percent. For example, a composition may be substantially free of added
oxidizing agent or of added persulfate according to the invention.

The term "about," as used herein, should generally be understood to
refer to both numbers in a range of numerals. Moreover, all numerical
ranges herein should be understood to include each whole integer
within the range.

The term "substantial," as used herein, means at least about 75%,
preferably at least about 90%, more preferably at least about 95%,
most preferably at least about 99%.

The term "valence state," as used herein, should be understood to
refer to the charge on a given ion or to the charge that may be
assigned to a given ion based on its electronic state.

The term "wound," as used herein, should be understood to refer to a
cut, laceration, abrasion, puncture or the like, especially in or on
the skin.

The terms "inhibit," "inhibiting," or "inhibits," as used herein when
referring to growth of an item, should be understood to refer to the
act of stopping that growth, whether permanently or temporarily, or of
reducing the rate of that growth, either permanently or temporarily.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

The tetrasilver tetroxide compounds mentioned in the background are
one type of electron active compound having multivalent cations in its
crystal lattice. Various additional electron active compounds have now
also been identified, as well as methods for making and using the same
for treating various pathogenic and non-pathogenic conditions or
disorders. It has now been discovered that such electron active
compounds, and compositions, beneficially enhance or facilitate the
growth of skin. Thus, the compounds, compositions, and methods and
articles using same, are advantageously used to treat or manage
various conditions that require rapid soft tissue growth, such as burn
therapy and skin grafts. The compounds and compositions of the
invention can increase the rate at which soft tissue grows, or heals,
compared to the ordinary rate in the absence of the invention. It is
also believed that the compounds and compositions of the invention
heal burns and other such conditions more rapidly and more
comprehensively than conventional burn treatments, such as silver
nitrate, silver sulfadiazine, and monovalent silver oxide (Ag.sub.2
O).

The electron active compounds of the present invention are believed to
have unique crystal structures in that, in the case of the metal
oxides, there are generally atoms of the same element in the crystal
that have at least two different valences, typically at least one
lower-valent metal cation and at least one higher-valent metal cation,
for example, such as Co(II) and Co(III), respectively. Exemplary
electron active metal oxide compounds according to the invention
include, but are not limited to, Ag(I,III), Co(II,III), Pr(III,IV),
Bi(III,V), Fe(II,III), Mn(II,III), and Cu(I,III) oxides. As discussed
below, pharmaceutical compositions including one or more of such oxide
compounds are useful for treating various conditions. The composition
of such exemplary electron active metal oxides is shown in tabular
form below:

Metal Lower-valent Higher-valent e Formula cations ion # ion # 2
Ag.sub.4 O.sub.4 Ag(I,III) Ag.sup.+ 2 Ag.sup.+3 2 1 Co.sub.3 O.sub.4
Co(II,III) Co.sup.+2 1 Co.sup.+3 2 2 Pr.sub.6 O.sub.11 Pr(III,IV)
Pr.sup.+3 2 Pr.sup.+4 4 2 Bi.sub.2 O.sub.4 Bi(III,V) Bi.sup.+3 1
Bi.sup.+5 1 1 Fe.sub.3 O.sub.4 Fe(II,III) Fe.sup.+2 1 Fe.sup.+3 2 1
Mn.sub.3 O.sub.4 Mn(II,III) Mn.sup.+2 1 Mn.sup.+3 2 2 Cu.sub.4 O.sub.4
Cu(I,III) Cu.sup.+ 2 Cu.sup.+3 2 e-total number of electrons believed
to be exchanged; #-number of particular ion type per formula unit.

Without being bound to theory, it is believed that the electron active
compounds operate against pathogens by transferring electrons between
their lower-valent ions and their higher-valent ions in the crystal,
thereby contributing to the death of pathogens by traversing their
cell membrane surface. It would seem that this, in effect,
"electrocutes" the pathogens. While these compounds have also been
discovered to be suitable for use in the prevention, treatment, and
management of other non-pathogenic conditions and disorders, such as
autoimmune disorders, circulatory disorders, neurological disorders,
and the like, the mechanism by which such conditions or disorders are
prevented, treated, or managed has not yet been fully understood. In
any event, the electrons in pathogens are believed to be perturbed
from their balanced crystals by such labile groups as NH, NH.sub.2, S--
S, and SH, which can be present, for example, in a pathogen cell
membrane. It is believed, however, that normal cells will not be
significantly affected because they do not proliferate rapidly enough
to expose these labile bonds sufficiently for the bonds to be
substantially affected.

The crystals in the electron active compounds are not believed to be
disturbed unless more stable complexes are formed with ligands, for
example, such as those comprising a pathogen cell membrane surface in
a dynamic state. Indeed, the end result of electron transfer, which is
a redox reaction, results in the lower-valent metal ions being
oxidized to one valence state higher and the higher-valent metal ions
being reduced to one valence state lower. In one embodiment, the
oxidation of the lower-valent metal ions and the reduction of the
higher-valent metal ions both result in ions having the same oxidation
state. Examples of such an embodiment occur when the valence
difference between the metal ions in the electron active molecular
crystal is 2 and such examples include, but are not limited to,
Ag(I,III), Bi(III,V), and Cu(I,III) oxides. In another embodiment, the
oxidation of the lower-valent metal ions and the reduction of the
higher-valent metal ions ?result in ions having opposite oxidation
states (e.g., ions with a +2 valence state are oxidized to +3, while
the ions with a +3 valence state are reduced to +2). Examples of such
an embodiment occur when the valence difference between the metal ions
in the electron active molecular crystal is 1 and such examples
include, but are not limited to, Co(II,III), Fe(II,III), Mn(II,III),
and Pr(III,IV) oxides.

The metal ion of certain electron active compounds may exhibit a
distinct affinity for certain elements of ligands, for example, such
as sulfur, oxygen, or nitrogen, particularly when present in a
pathogen's cell membrane. In many cases, the metal ion will not merely
bind to these elements, but will actually form chelate complexes with
their ligands. The classic example of this is Ag(I,III) oxide, the
monovalent silver ion of which has an affinity for sulfur and nitrogen
and the oxidized/reduced divalent ion of which forms chelate complexes
with, for example, mercapto or amino groups. Thus, the electron active
compound attraction for the cell membrane surfaces, for example, of
pathogens, is believed to be driven by powerful electrostatic forces.

Without being bound by theory, the electron exchange may be depicted,
for example, by the following series of redox half reactions:

metal(III,IV) metal(III,V) metal(I,III) oxides metal(II,III) oxides
oxides oxides Ag.sup.+ - e = Ag.sup.+2 Co.sup.+2 - e = Co.sup.+3
Pr.sup.+3 - e = Pr.sup.+4 Bi.sup.+3 - e = Bi.sup.+4 Ag.sup.+3 + e =
Ag.sup.+2 Co.sup.+3 + e = Co.sup.+2 Pr.sup.+4 + e = Pr.sup.+3 Bi.sup.
+5 + e = Bi.sup.+4 Cu.sup.+ - e = Cu.sup.+2 Fe.sup.+2 - e = Fe.sup.+3
Cu.sup.+3 + e = Cu.sup.+2 Fe.sup.+3 + e = Fe.sup.+2 Mn.sup.+2 - e =
Mn.sup.+3 Mn.sup.+3 + e = Mn.sup.+2

For each redox reaction, there is believed to be an electromotive
force, which is the voltage potential when oxidizing the higher-valent
ion in the metal oxide crystal. This is denoted herein as EMF.sup.OX.
In addition to the electromotive force of oxidation, there is believed
to be an associated reduction reaction involving the lower-valent ion
in the metal oxide crystal. This reduction reaction may be represented
simply, as tabulated above, or may represent the interaction with, for
example, a ligand present on a pathogen cell membrane surface, such as
one containing sulfur or nitrogen. Associated with the reduction
reaction is another electromotive force, or voltage potential when
reducing the lower-valent ion. This is denoted herein as EMF.sup.RE.

When the metal ions of the electron active metal oxide interact with,
for example, a sulfur-containing ligand, the affinity of the metal ion
for sulfur affects EMF.sup.RE. The stability of a particular metal
sulfide is an approximation of the affinity of a metal ion for sulfur.
The following approximate association constants for sulfides indicate
the trend in relative affinity of each metal ion for sulfur:

Ag(I) 49 Cu(I) 47 Co(II) 26 Fe(II) 19 Mn(II) 15

In general, the more stable the compound, the more negative its
reduction potential in the reduction reaction, for example, in the
case of elemental silver:

In the case of tetrasilver tetroxide, there is a reduction reaction
where Ag(I) is oxidized and an oxidation reaction where Ag(III) is
reduced, as follows:

Ag.sup.+ -e+S.sup.-2 -e.fwdarw.AgS EMF.sup.RE =-0.90

The voltage that is discharged from a redox reaction of the electron
active metal oxides of the present invention, which voltage is denoted
herein as the "electrocution voltage," is the combination of the
oxidation and reduction potentials (i.e., EMF.sup.OX -EMF.sup.RE). In
the case of tetrasilver tetroxide, the "electrocution voltage" is 2.92
volts. The oxidation potentials, EMF.sup.OX, of exemplary metal oxides
according to the present invention are tabulated below:

Formula Metal cations EMF.sup.OX Ag.sub.4 O.sub.4 Ag(I,III) 2.02
Co.sub.3 O.sub.4 Co(II,III) 1.81 Pr.sub.6 O.sub.11 Pr(III,IV) 2.86
Bi.sub.2 O.sub.4 Bi(III,V) 1.59 Fe.sub.3 O.sub.4 Fe(II,III) 0.77
Mn.sub.3 O.sub.4 Mn(II,III) 1.54 Cu.sub.4 O.sub.4 Cu(I,III) 1.80

As noted from the above table, praseodymium-, cobalt-, and copper-
based oxides are believed to be stronger antipathogenic agents or to
form better pharmaceutical compositions than manganese-, bismuth-, and
iron- based oxides, and in one embodiment they are preferred for this
reason. Nevertheless, in certain cases, iron exhibits stronger
antipathogenic characteristics, particularly antimicrobial
characteristics, compared to manganese.

Another factor, however, particularly in antipathogenic or
antimicrobial efficacy, can be the sulfur/nitrogen composition, for
example, of cell membranes. For example, Staphylococcus aureus
bacteria, in a culture having a cell density of 30,000 CFU/mL, exhibit
significant mortality from exposure to 100 ppm of Bi(III,V) oxide for
about 10 minutes, but no significant mortality from exposure to the
same concentrations of Fe(II,III) and Mn(II,III) oxides for the same
contact time. This result might be explained by the far greater
stability of bismuth(III) sulfide, and thus the far greater affinity
of bismuth(III) for sulfur, than either of the iron(II) or
manganese(II) analogs.

The electron active metal oxide compounds and compositions of the
present invention may be used in any form which sufficiently retains
their antipathogenic character, or other non-pathogenic ability, to
prevent, treat, or manage one or more of the conditions noted herein.

These compounds or compositions may be used as antipathogenic agents,
such as antimicrobial, antibacterial, antiviral, or anti-algal agents,
or a combination thereof. In another embodiment, the compounds or
compositions may be used for preventing, treating, and/or managing
various conditions that are non-pathogenic. For example, non-
pathogenic conditions are believed to include certain autoimmune
disorders, neurological disorders, and circulatory disorders. While
the exact mechanism of the activity of such compounds or compositions
is not described herein, nonetheless, suitable prevention, treatment,
and/or management of such non-pathogenic conditions may be obtained by
administering the compounds or compositions of the invention as
described herein and as will be readily apparent to one of ordinary
skill in the art.

The compositions and methods of the invention advantageously prevent,
treat, or manage dermatological diseases or conditions. The conditions
against which the electron active compounds, such as metal oxides, of
the present invention have utility include, but are not limited to,
Madura foot, actinomycosis, oral actinomycosis, anthrax, food
poisoning, botulism, wound infections, pseudomembranous colitis,
colitis, gas gangrene, gangrene, tetanus, diphtheria, pharyngeal
diphtheria, pleomorphic laryngeal diphtheria, cutaneous diphtheria,
endocarditis, bacteremia, urinary tract infections, listerosis,
meningitis, miscarriage, narcodiosis, acne, skin lesions, abscesses,
toxic shock syndrome, prosthesis contamination, dental caries, plaque,
gum disease, gingivitis, subacute endocarditis, bacterial pneumonia,
otitis, sinusitis, cat scratch fever, septicemia, abdominal and pelvic
abscesses, Oroya fever, systemic Oroya fever, verruga peruana,
cutaneous verruga peruana, whooping cough, Lyme disease, epidemic
relapsing fever, brucellosis, granuloma inguinale granulomatic,
donovanosis, gastroenteritis, nosocomial infections, tularemia,
bacterial vaginitis, urethritis, bacterial conjunctivitis, chancroid,
otitis media, chronic gastritis, peptic ulcer, diarrhea, Legionnaires'
disease, leptospirosis, gonorrhea, arthritis, periodontal disease,
salmonellosis, typhoid fever, shigellosis, rat bite fever,
pharyngitis, scarlet fever, syphilis, cholera, Asiatic cholera,
Yersina arthritis, bubonic plague, chronic pulmonary disease, Hansen's
disease, leprosy, tuberculosis, dermal tuberculosis, psittachosis,
omithosis, conjunctivitis, trachoma, lymphogranuloma venereum, genital
tract infections, Q fever, primary atypical pneumonia, rickettsial
pox, typhus, epidemic typhus, Rocky Mountain spotted fever,
tsutsugamushi fever, nongonococcal urethritis, human erlichiosis,
meningococcal meningitis, skin infections, corneal infections,
external ear infections, candidiasis, monoiliasis, thrush, candidosis,
mucositis, bacteremia, hepatitis, hepatitis A, hepatitis B, hepatitis
C, hepatitis E, coccidiomycosis, lymphadenitis, balantidiasis
cryptosporidosis, amoebiasis, amoebic dysentery, giardiasis, giardia
enteritis, leishmaniasis, Kala-azar, malaria, toxoplasmosis,
trypanosomiasis, Chagas disease, African sleeping sickness, dengue,
Japanese encephalitis, Rift Valley fever, Ebola hemorrhagic fever,
Venezuelan hemorrhagic fever, hantavirus pulmonary syndrome,
hemorrhagic fever with renal syndrome, cytomegalovirus infection,
poliomyelitis, West Nile virus disease, influenza, measles, condyloma,
encephalitis, ankylosing spondylitis, arteritis, inflammatory bowel
disease, polyarteritis nodosa, rheumatic fever, systemic Lupus
erythematosus, Alzheimer's disease, multiple sclerosis, osteoporosis,
Crohn's disease, strep throat, yellow fever, eczema, psoriasis,
dermatitis, disease-induced skin ulcers, undefined tropical diseases,
shingles, rashes, heat rashes, bedsores, cold sores, blisters, boils,
herpes simplex, acne, pimples, skin chafing, skin cracking, itchiness,
skin peeling, warts, one or more symptoms thereof, or any combination
thereof. In another embodiment, the condition includes HIV (AIDS), or
one or more symptoms. It should be understood that the invention
includes the use of the compounds or compositions to prevent, treat,
or manage each of these conditions individually or multiple conditions
concurrently or sequentially. Thus, the prevention, treatment, or
management of each condition should be understood as a separate
embodiment.

The pathogens which may be killed by, or the growth or proliferation
of which may be halted, diminished, or inhibited by, the electron
active metal oxides of the present invention include, but are not
limited to, gram-positive bacilli and cocci; gram-negative bacilli and
cocci; acid-fast bacteria; other bacteria; fungi; parasitic microbes,
e.g., protozoa; and viruses.

Examples of gram-positive bacilli and cocci include, but are not
limited to, Actinomedurae, Actinomyces israelii, Bacillus anthracis,
Bacillus cereus, Clostridium botulinum, Clostridium difficile,
Clostridium perfringens, Clostridium tetani, Corynebacterium,
Enterococcus faecalis, Listeria monocytogenes, Nocardia,
Propionibacterium acnes, Staphylococcus aureus, Staphylococcus
epiderm, Streptococcus mutans, Streptococcus pneumoniae, and
combinations thereof.

Examples of gram-negative bacilli and cocci include, but are not
limited to, Afipia felis, Bacteriodes, Bartonella bacilliformis,
Bortadella pertussis, Borrelia burgdorferi, Borrelia recurrentis,
Brucella, Calymmatobacterium granulomatis, Campylobacter, Escherichia
coli, Francisella tularensis, Gardnerella vaginalis, Haemophilius
aegyptius, Haemophilius ducreyi, Haemophilius influenziae, Heliobacter
pylori, Legionella pneumophila, Leptospira interrogans, Neisseria
meningitidia, Porphyromonas gingivalis, Providencia sturti,
Pseudomonas aeruginosa, Salmonella enteridis, Salmonella typhi,
Serratia marcescens, Shigella boydii, Streptobacillus moniliformis,
Streptococcus pyogenes, Treponema pallidum, Vibrio cholerae, Yersinia
enterocolitica, Yersinia pestis, and combinations thereof.

Examples of acid-fast bacteria include, but are not limited to,
Myobacterium avium, Myobacterium leprae, Myobacterium tuberculosis,
and combinations thereof.

Examples of other bacteria not falling into the other three categories
include, but are not limited to, Bartonella henseiae, Chlamydia
psittaci, Chlamydia trachomatis, Coxiella burnetii, Mycoplasma
pneumoniae, Rickettsia akari, Rickettsia prowazekii, Rickettsia
rickettsii, Rickettsia tsutsugamushi, Rickettsia typhi, Ureaplasma
urealyticum, Diplococcus pneumoniae, Ehrlichia chafensis, Enterococcus
faecium, Meningococci, and combinations thereof.

Examples of fungi include, but are not limited to, Aspergilli,
Candidae, Candida albicans, Coccidioides immitis, Cryptococci, and
combinations thereof.

Examples of parasitic microbes include, but are not limited to,
Balantidium coli, Cryptosporidium parvum, Cyclospora cayatanensis,
Encephalitozoa, Entamoeba histolytica, Enterocytozoon bieneusi,
Giardia lamblia, Leishmaniae, Plasmodii, Toxoplasma gondii,
Trypanosomae, trapezoidal amoeba, and combinations thereof.

Examples of viruses include, but are not limited to, Arboviruses,
Ebola virus, Guanarito virus, Hanta virus, Hantaan virus, Hepatitis A,
Hepatitis B, Hepatitis C, Hepatitis E, other Hepatitis viruses, Herpes-
type viruses, Poliovirus, West Nile virus, Echo virus, and
combinations thereof.

The antipathogenic or non-pathogenic compositions of the present
invention may optionally further include the use of one or more
additional therapeutic agents known to treat a condition, or a symptom
thereof. Examples of such additional therapeutic agents include, but
are not limited to, chelating agents, vitamins, minerals, silica
hydride microclusters, analgesics, SambucolTM, aspirin, and the like.

The administration of one or more active ingredients and/or optional
therapeutic agent(s), in accordance with the methods of the invention
may occur together, concurrently but separately, sequentially, or a
combination thereof. The optional additional therapeutic agent is
generally a compound other than an electron active metal oxide
compound.

The antipathogenic or antimicrobial performance of certain metal
oxides may be improved or enhanced by the presence of an oxidizing
agent. This is particularly the case when the metal oxide compounds or
compositions are present in low amounts, i.e., typically less than 45
ppm, and more commonly when present in an amount less than about 40
ppm, based on the weight of the composition. In such situations, an
oxidizing agent may be included in certain compositions of the
invention in small amounts when the compositions are administered by
certain routes. In such an embodiment, the oxidizing agent includes a
peroxy acid salt, preferably a Group I salt of a persulfate, more
preferably potassium persulfate. In another embodiment, the oxidizing
agent includes the same peroxy acid salt which was present as a
starting material in the reaction to form the particular electron
active metal oxide. The oxidizing agent may advantageously be present
in the composition in amounts from about 1 ppm to 500 ppm, based on
the weight of the composition. In alternate embodiments, there may be
from about 5 ppm to 200 ppm or from about 10 ppm to 100 ppm of
oxidizing agent, based on the weight of the composition.

It is believed that the additional presence of certain types or
amounts of oxidizing agent(s) may tend to irritate the skin,
particularly when the compound or composition including metal oxide(s)
is present in large amounts, such as greater than 50 ppm, based on the
weight of the composition. In one embodiment, as more compound or
composition is administered, a correspondingly smaller amount of
undesirable oxidizing agent is required. Thus, in some embodiments, it
has been found that the additional oxidizing agent is unnecessary and
in fact undesirable for the purpose of treating certain conditions
described herein, since the additional oxide may have or contribute to
an undesirable side effect, for example, such as skin irritation when
applied topically. For those embodiments, the compositions minimize
the amount of additional oxidizing agent, such as persulfate, or are
substantially or completely free of added persulfates or other
oxidizing agents.

Certain of the electron active metal oxides may be black in color,
such that care must be taken when formulating suitable topical
pharmaceutical compositions according to the invention to inhibit
blackening or superficial discoloration of the skin. Without being
bound by theory, it is believed that larger amounts of such
compositions promote increased superficial discoloration. Thus, in one
embodiment, the pharmaceutical compositions preferably have an
insufficient amount of metal oxide composition to cause visible skin
discoloration.

Additionally, it was found by rigorous testing that certain silver
tetroxide-containing compositions were comparatively non-toxic
compared to silver salts, such as conventional formulations of silver
nitrate, silver sulfadiazine, and benzoyl peroxide. Since these silver
tetroxide compositions were effective at certain ppm concentrations in
killing pathogens in nutrient broth and for water treatment,
commercial concentrates were formulated with 2% of the tetrasilver
tetroxide. For acceptance of the oxide in commerce, for which EPA
registration No. 3432-64 was obtained, it was necessary for the Ag.sub.
4 O.sub.4 to undergo a series of toxicity tests. A 3% concentrate was
used and evaluated by a certified laboratory employing good laboratory
practice (GLP) according to the Code of Federal Regulations for this
purpose. The results were as follows:

Acute Oral Toxicity LD.sub.50 Greater than 5,000 mg/Kg Acute Dermal
Toxicity LD.sub.50 Greater than 2,000 mg/Kg Primary Eye Irritation
Mildly irritating Primary Skin Irritation No irritation Skin
Sensitization Non-Sensitizing

Subsequent evaluations conducted according to the invention showed
that unless persons were prone to silver allergies, the pure
tetrasilver tetroxide compositions according to the invention could be
applied to the skin without any ill effects or evidence of irritation,
despite the fact that the compositions of the invention can be a
powerful oxidizing agent.

Where the electron active compositions according to the invention are
applied to the skin, they may be combined with a carrier in an amount
from about 1 ppm to 500,000 ppm, more preferably from about 50 ppm to
250,000 ppm, of the electron active metal oxide composition, based on
the weight of the composition. In various embodiments, the
compositions are provided in amounts from about 100 ppm to 100,000
ppm, from about 500 ppm to 70,000 ppm, from about 5,000 ppm to 50,000
ppm, or from about 10,000 ppm to 40,000 ppm, based on the weight of
the composition. In one preferred embodiment, the compositions are
formulated with about 25,000 ppm to 35,000 ppm of metal oxide, based
on the weight of the composition. It will be readily understood by
those of ordinary skill in the art that the ppm concentration of
electron active compound(s), such as metal oxide, in the composition
is based on the total weight of the composition.

When treating or managing conditions that require skin growth, such as
burn therapy or skin graft management or treatment, a preferred
embodiment employs amounts of about 0.1 to 10 percent by weight, about
0.25 to 5 percent by weight, or about 2 to 4 percent by weight of the
compounds or compositions of the invention. The compositions, when
applied topically, can be applied to the skin about 1 to 3 times per
day until the condition is suitably cured or satisfactorily
controlled. In one embodiment, the composition may generally be
topically applied at a dosage level of from about 1 mg to 1000 mg per
cm.sup.2 of skin surface, preferably about 10 mg to 500 mg per cm.sup.
2 of skin surface. When applied topically, a preferred carrier
includes petroleum jelly, such as white petroleum jelly. For example,
a suitable white petroleum jelly is available from Penreco of Houston,
Tex.

Most of the metal oxide compounds, for example, for use according to
the invention are commercially available from various sources.
Tetrasilver tetroxide compositions for use according to the invention
have been commercially sold under the poorly named "Ag(II) OXIDE"
tradename. They may be obtained from Aldrich Chemical Co., Inc.,
having a place of business in Milwaukee, Wis. The chemical synthesis
of tetrasilver tetroxide compounds can also be performed according to
the method described on page 148 in M. Antelman, "Anti-Pathogenic
Multivalent Silver Molecular Semiconductors," Precious Metals, vol.
16:141-149 (1992) by reacting silver nitrate with potassium
peroxydisulfate according to the following equation in alkali
solutions:

To the extent necessary to understand the present invention, the
disclosure of Antelman is hereby incorporated herein by express
reference thereto.

Tetracopper tetroxide, also referred to herein as Cu(I,III) oxide or
CU.sub.4 O.sub.4, may be prepared as follows.

Suitable copper-based starting materials for this reaction include at
least one copper(I)-containing material. In one embodiment, a water
soluble copper(I) salt can be used. Typically, a water soluble
copper(I) salt can be prepared by dissolving an inorganic copper(I)
compound, for example, such as cuprous oxide, in an appropriate acid,
for example, an organic acid, such as acetic acid. Since soluble
copper(I) salts are not readily commercially available at the present
time, however, a non-solvated inorganic copper(I) compound, such as
cuprous oxide itself, can be used as the copper(I)-containing starting
material. In addition, other copper(I)-containing materials, either
inorganic, such as a copper(I) oxide, or organic, such as an
organometallic copper(I) compound, or both, may be used, where the
copper(I)-containing material(s) are sufficiently soluble in an
aqueous or organic solution to allow reaction with other materials to
form an electron active copper oxide compound.

The copper(I)-containing starting material is combined with an aqueous
caustic solution. This caustic solution preferably contains two
components: a strong caustic base and a peroxy acid salt. Examples of
suitable strong caustic bases include Group I and Group II hydroxides,
preferably sodium hydroxide or potassium hydroxide. Examples of
suitable peroxy acid salts include Group I salts of persulfates,
preferably potassium persulfate.

The copper-based starting material is typically the limiting reagent
in such a preparation. The ratio of each of the components in the
caustic solution to that of the copper-based starting material is
theoretically set by the stoichiometry of the particular reaction. In
one preferred embodiment, there is a relative molar excess, i.e., an
amount more than stoichiometrically necessary, of each of the
components in the caustic solution with respect to the copper-based
starting material. When a strong caustic base and a peroxy acid salt
are present in the caustic solution, the relative molar excesses of
the components may be at least about 50% and at least about 10%,
respectively, preferably at least about 100% and at least about 20%,
respectively, more preferably, at least about 250% and at least about
40%, respectively, most preferably at least about 500% and at least
about 75%, respectively.

Generally, the reactants may be added together in any manner that
comports with typical laboratory procedure. In one embodiment, the
copper(I)-containing starting material is placed in a reactor, to
which the strong caustic base and the peroxy acid salt are added, each
typically in their own solutions. The solution containing the
reactants is then typically heated to a temperature sufficient to
activate a reaction, preferably sufficient to activate a reaction with
no major undesirable side reactions or other undesirable effects, more
preferably above about 80.degree. C., most preferably about 90.degree.
C. to 95.degree. C. The solution is heated for a time sufficient to
facilitate the reaction, preferably to provide substantial completion
of the reaction, preferably for at least about 5 minutes, more
preferably for at least about 15 minutes, after which time the
solution is allowed to cool or is cooled, preferably to below about
45.degree. C., more preferably to about room temperature.

The color change of the solution, from its original color, red, to a
color indicating a reaction has occurred, in this case black, may
occur at the heated temperature or during or after cooling.

The purification and isolation of the desired product can be
accomplished by any suitable method available to those of ordinary
skill in the art. In the majority of situations, the desired reaction
product is primarily a solid, but may be dissolved or dispersed in at
least part of the solution. In one preferred embodiment, the solution
is carefully decanted off, and then the remaining product is washed
multiple times with distilled water, before being sufficiently dried.
In another preferred embodiment, the solution is vacuum filtered to
remove the filtrate, and the remaining product is sufficiently dried.

The yield of solid tetracopper tetroxide material, based on the
reactants, is typically at least about 10%, preferably at least about
45%, more preferably at least about 75%, most preferably at least
about 80%.

In addition, Fe(II,III) oxide and Mn(II,III) oxide are commercially
available from Aldrich Company of Milwaukee, Wis., and Co(II,III)
oxide and Pr(III,IV) oxide are commercially available from Noah
Technologies of San Antonio, Tex. Also, Bi(III,V) oxide synthetic
routes are detailed and reviewed in Gmelins Handbuch Der Anorganischen
Chemie, vol. 16:642 (1964), and the oxide is available commercially
from City Chemicals of New York, N.Y.

The magnitude of a prophylactic or therapeutic dose of electron active
composition(s), or a derivative thereof, in the acute or chronic
management of diseases and disorders described herein will vary with
the severity of the condition to be prevented, treated, or managed and
the route of administration. For example, oral, mucosal (including
rectal and vaginal), parenteral (including subcutaneous,
intramuscular, bolus injection, and intravenous, such as by infusion),
sublingual, transdermal, nasal, buccal, and like may be employed. In
one embodiment, a patient may gargle using the composition of the
present invention. Dosage forms include tablets, troches, lozenges,
dispersions, suspensions, suppositories, solutions, capsules, soft
elastic gelatin capsules, patches, and the like. The dose, and perhaps
the dose frequency, will also vary according to the age, body weight,
and response of the individual patient. Suitable dosing regimens can
be readily selected by those of ordinary skill in the art with due
consideration of such factors. In general, the total daily dosage for
the conditions described herein, is from about 0.1 mg to 1,000 mg of
the active ingredient, i.e., one of the metal oxides described herein,
or a derivative thereof. In another embodiment, the daily dosage can
be from about 1 mg to 500 mg, while in another embodiment, the daily
dosage can be from about 2 mg to 200 mg of the metal oxide
composition. A unit dosage can include, for example, 30 mg, 60 mg, 90
mg, 120 mg, or 300 mg of metal oxide composition. Preferably, the
active ingredient is administered in single or divided doses from one
to four times a day, such as by topical administration. In another
embodiment, the compositions are administered by an oral route of
administration. The oral dosage forms may be conveniently presented in
unit dosage forms and prepared by any methods available to those of
ordinary skill in the art of pharmacy.

In managing the patient, the therapy may be initiated at a lower dose,
e.g., from about 1 mg, and increased up to the recommended daily dose
or higher depending on the patient's global response. It is further
recommended that children, patients over 65 years, and those with
impaired renal or hepatic function, initially receive low doses when
administered systemically, and that they be titrated based on
individual response(s) and blood level(s). It may be necessary to use
dosages outside these ranges in some cases, as will be apparent to
those of ordinary skill in the art. Furthermore, it is noted that the
clinician or treating physician will know how and when to interrupt,
adjust, or terminate therapy in conjunction with individual patient
response.

Any suitable route of administration may be employed for providing the
patient with an effective dosage of electron active metal oxide, or a
derivative thereof. The most suitable route in any given case will
depend on the nature and severity of the condition being prevented,
treated, or managed. In one embodiment where burns or skin grafts are
being treated or managed, the compounds or compositions may be
administered topically.

In practical use, the electron active compound, such as a metal oxide,
or a derivative thereof, can be combined as the active ingredient in
intimate admixture with a pharmaceutical carrier medium according to
conventional pharmaceutical compounding techniques. The carrier may
take a wide variety of forms and may include a number of components
depending on the form of preparation desired for administration. The
compositions of the present invention may include, but are not limited
to, suspensions, solutions and elixirs; aerosols; or carriers,
including, but not limited to, starches, sugars, microcrystalline
cellulose, diluents, granulating agents, lubricants, binders,
disintegrating agents, and the like.

Suitable forms in which the electron active compounds or compositions
of the present invention may be used include, but are not limited to,
powder, granule, flake, solution, suspension, emulsion, slurry,
aerosol spray, gel, paste, and combinations thereof. In one preferred
embodiment, the form is a powder or solution. When the electron active
compounds are in the form of a solution, the solution may be aqueous,
non-aqueous, or a combination thereof, preferably at least partially
aqueous, more preferably substantially aqueous. In a preferred
embodiment, the metal oxides are in an aqueous solution.

The compositions of the invention may be applied topically, e.g.,
either directly as a powder, powder crystals, or granules, or in other
non-sprayable or sprayable forms. Non-sprayable forms can be semi-
solid or solid forms including a carrier indigenous to topical
application and preferably having a dynamic viscosity greater than
that of water. Suitable formulations include, but are not limited to,
suspensions, emulsions, creams, ointments, powders, liniments, salves
and the like. If desired, these may be sterilized or mixed with any
available auxiliary agents, carriers, or excipients, e.g.,
thixotropes, stabilizers, wetting agents, and the like. One or more
thixotropic agents can be included in types and amounts sufficient to
increase adhesion of topically applied compositions of the invention
to the skin, so as to inhibit or prevent runoff or other loss of the
composition from the treatment zone on the skin. Preferred vehicles
for non-sprayable topical preparations include ointment bases, e.g.,
polyethylene glycol-1000 (PEG-1000); conventional ophthalmic vehicles;
creams; and gels, as well as petroleum jelly and the like. In one more
preferred embodiment, the carrier includes a petroleum jelly. In
another preferred embodiment, the carrier is formulated as a cream,
gel, or lotion. In another preferred embodiment, the carrier is 3
weight percent active ingredient, 36 weight percent heavy mineral oil,
47 weight percent petroleum jelly, and 14 weight percent Tivawax P,
which is available from Tivian Laboratories, Inc., of Providence, R.I.
In yet another preferred embodiment, the composition may be a dry
powder, such as with 5 weight percent active ingredient and 95 weight
percent bismuth subgallate. These topical preparations may also
contain emollients, perfumes, and/or pigments to enhance their
acceptability for various usages.

The compositions may also be formulated for parenteral administration
by injection (subcutaneous, bolus injection, intramuscular, or
intravenous, such as by infusion), and may be dispensed in a unit
dosage form, such as a multidose container or an ampule. Compositions
of the electron active metal oxide, or a derivative thereof, for
parenteral administration may be in the form of suspensions,
solutions, emulsions, or the like, in aqueous or oily vehicles, and in
addition to the active ingredient, may contain one or more formulary
agents, such as dispersing agents, suspending agents, stabilizing
agents, preservatives, and the like.

In the case where an intravenous injection or infusion composition is
employed, a suitable dosage range can be, e.g., from about 0.5 mg (0.1
ppm) to about 1,000 mg (200 ppm) total dose, preferably from about 5
mg (1 ppm) to 400 mg (80 ppm). In one preferred embodiment, the total
dose can be from about 50 mg (10 ppm) to 200 mg (40 ppm). It should be
understood that any suitable amount of the composition according to
the invention may be administered if effective to prevent, treat, or
manage one or more conditions described herein.

Pharmaceutical compositions of the present invention may be orally
administered in discrete pharmaceutical unit dosage forms, such as
capsules, cachets, soft elastic gelatin capsules, tablets, or aerosols
sprays, each containing a predetermined amount of the active
ingredient, as a powder or granules, or as a solution or a suspension
in an aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion,
or a water-in-oil liquid emulsion. Such compositions may be prepared
by any of the methods of pharmacy, but all methods include the step of
bringing into association the active ingredient with the
pharmaceutically acceptable carrier which constitutes one or more
necessary ingredients. In general, the compositions are prepared by
uniformly and intimately admixing the active ingredient with liquid
carriers or finely divided solid carriers or both, and then, if
necessary, shaping the product into the desired presentation. Suitable
types of oral administration include oral solid preparations, such as
capsules or tablets, or oral liquid preparations. If desired, tablets
may be coated by standard aqueous or nonaqueous techniques.

For example, a tablet may be prepared by compression or molding,
optionally, with one or more accessory ingredients. Compressed tablets
may be prepared by compressing in a suitable machine the active
ingredient in a free-flowing form such as powder or granules,
optionally mixed with a binder, lubricant, inert diluent, granulating
agent, surface active agent, dispersing agent, or the like. Molded
tablets may be made by molding, in a suitable machine, a mixture of
the powdered compound moistened with an inert liquid diluent. In one
embodiment, each tablet, capsule, cachet, or gel cap contains from
about 0.5 mg to about 500 mg of the active ingredient, while in
another embodiment, each tablet contains from about 1 mg to about 250
mg of the active ingredient. The amount of active ingredient found in
the composition, however, may vary depending on the amount of active
ingredient to be administered to the patient.

The electron active compound(s), or a derivative thereof, may be
formulated as a pharmaceutical composition in a soft elastic gelatin
capsule unit dosage form by using conventional methods well known in
the art, such as in Ebert, Pharm. Tech, 1(5):44-50 (1977). Soft
elastic gelatin capsules have a soft, globular gelatin shell somewhat
thicker than that of hard gelatin capsules, wherein a gelatin is
plasticized by the addition of plasticizing agent, e.g., glycerin,
sorbitol, or a similar polyol. The hardness of the capsule shell may
be changed by varying the type of gelatin used and the amounts of
plasticizer and water. The soft gelatin shells may contain an
additional preservative, such as methyl- and propylparabens and sorbic
acid, to prevent the growth of fungi, although this is not essential
since the compounds and compositions of the invention provide anti-
fungal efficacy. Thus, in one embodiment, the invention includes a
compositions formulated as a gelatin shell with the composition of the
invention, e.g.,a metal oxide, completely free of added preservatives.
The active ingredient may be dissolved or suspended in a liquid
vehicle or carrier, such as vegetable or mineral oils, triglycerides,
surfactants such as polysorbates, or a combination thereof.

In addition to the common dosage forms set out above, the compounds of
the present invention may also be administered by controlled release
means, delivery devices, or both, as are well known to those of
ordinary skill in the art, such as those described in U.S. Pat. Nos.:
3,845,770; 3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533;
5,059,595; 5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; and
5,733,566, the disclosures of which are hereby incorporated herein by
express reference thereto. These pharmaceutical compositions can be
used to provide slow or controlled-release of the active ingredient
therein using, for example, hydropropylmethyl cellulose in varying
proportions to provide the desired release profile, other polymer
matrices, gels, permeable membranes, osmotic systems, multilayer
coatings, microparticles, liposomes, microspheres, or the like, or a
combination thereof. Suitable controlled-release formulations
available to those of ordinary skill in the art, including those
described herein, may be readily selected for use with the
compositions of the invention. Thus, single unit dosage forms suitable
for topical or oral administration, such as gels, lotions, cremes,
tablets, capsules, gelcaps, caplets, and the like, that are adapted
for controlled-release are encompassed by the present invention.

All controlled-release pharmaceutical products have a common goal of
improving drug therapy over that achieved by their non-controlled
counterparts. Ideally, the use of an optimally designed controlled-
release preparation in medical treatment is characterized by a minimum
of drug substance being employed to cure or control the condition in a
minimum amount of time. Advantages of controlled-release formulations
may include: 1) extended activity of the drug; 2) reduced dosage
frequency; and 3) increased patient compliance.

Most controlled-release formulations are designed to initially release
an amount of drug that promptly produces the desired therapeutic
effect, and gradual and continual release of other amounts of drug to
maintain this level of therapeutic effect over an extended period of
time. In order to maintain this constant level of drug in the body,
the drug should be released from the dosage form at a rate that will
replace the amount of drug being metabolized and excreted from the
body.

The controlled-release of the active ingredient may be stimulated by
various inducers, for example pH, temperature, enzymes, water, or
other physiological conditions or compounds. The term "controlled-
release component" in the context of the present invention is defined
herein as a compound or compounds, including polymers, polymer
matrices, gels, permeable membranes, liposomes, microspheres, or the
like, or a combination thereof, that facilitates the controlled-
release of the active ingredient (e.g., tetrasilver tetroxide or
another metal oxide) in the pharmaceutical composition.

The skin-growth-enhancing compositions for use in the present
invention include electron active metal oxides, or a derivative
thereof, as the active ingredient, and may also contain a
pharmaceutically acceptable carrier, and optionally, other therapeutic
ingredients. Suitable derivatives include any available
"pharmaceutically acceptable salts," which refer to a salt prepared
from pharmaceutically acceptable non-toxic acids including inorganic
acids, organic acids, solvates, hydrates, or clathrates thereof.
Examples of such inorganic acids are nitric, sulfuric, lactic,
glycolic, salicylic, and phosphoric. Appropriate organic acids may be
selected, for example, from aliphatic, aromatic, carboxylic and
sulfonic classes of organic acids, examples of which are formic,
acetic, propionic, succinic, camphorsulfonic, citric, fumaric,
gluconic, isethionic, lactic, malic, mucic, tartaric, para-
toluenesulfonic, glycolic, glucuronic, maleic, furoic, glutamic,
benzoic, anthranilic, salicylic, phenylacetic, mandelic, embonic
(pamoic), methanesulfonic, ethanesulfonic, pantothenic,
benzenesulfonic (besylate), stearic, sulfanilic, alginic,
galacturonic, and the like. Particularly preferred acids are lactic,
glycolic, and salicylic acids. The pharmaceutically acceptable salts
preferably do not include halide-containing salts when tetrasilver
tetroxide is present, as these salts are believed to facilitate
breakdown of the oxide lattice present in the silver oxide
compositions of the invention.

EXAMPLES

These and other aspects of the present invention may be more fully
understood with reference to the following non-limiting examples,
which are merely illustrative of the preferred embodiment of the
present invention, and are not to be construed as limiting the
invention, the scope of which is defined by the appended claims.

Example 1

Method of Treating Diabetes-Induced Foot Ulcers According to Invention

Twenty eight patients in the age group ranging from 45 to 65 having
diabetes-induced foot ulcers were arranged in two groups. All of the
patients were taking insulin injections and were diagnosed as Type I
insulin dependent. Moreover, all of the patients had presented the
diabetic foot condition for at least 10 days prior to treatment

Group I included fourteen patients where culture swabs of the
ulcerated skin indicated the presence of bacteria (infection). Group
II included fourteen patients where culture swabs of the ulcerated
skin did not indicate the presence of abnormal amounts of bacteria (no
infection).

The patients in each group were treated by applying 200 mg of a
petroleum jelly containing 3 wt % tetrasilver tetroxide twice daily to
the ulcerated sores for a 30-day period. Daily evaluations of the skin
condition were conducted by a dermatologist.

Summary of Results

Group I: Within 48 hours of the onset of treatment, the sores on the
feet of all patients began to dry out. After 72 hours, the ulcers on
all patients started to heal at the borders. By the fourth day,
inflammation of the diseased tissue eased, and by the sixth day the
ulcers were completely dry with no surface secretions. By the tenth
day, the ulcers on all patients feet had completely disappeared. Lab
tests indicated no sign of infection on the feet of any patient by the
tenth day.

Group II: Within 24 hours of the onset of treatment the sores on the
feet of all patients began to dry out and heal at the borders with no
secretion. By the third day, the sores on all patients were covered
with new healthy tissue. By the tenth day, the ulcers had healed and
completed the process of forming scar tissue by 80%. At day 14 of the
treatment, all of the ulcers were 100% healed with no sign of
infection.

Continuous monitoring of both groups over the 30-day period indicated
no reappearance of the ulcers.

The above tests demonstrated that tetrasilver tetroxide treatment was
effective in both curing infections associated with diabetes-induced
ulcers and healing the ulcers themselves. Without being bound by
theory, it is believed that the active tetroxide compositions of the
present invention accelerated the neovascularization process of the
affected tissue and facilitated the treatment.

Example 2

Effect of Compositions of Invention on Burn Therapy

Fourteen (14) patients ranging in age from 11 to 38 years old were
diagnosed as having tissue injury caused by thermal contact. These
patients were arranged in two groups, and all previous treatments were
removed before any new treatments were applied.

Group I: 7 patients were diagnosed as having second degree burns, 20%
BSA, of moderate severity on the hands and arms. These patients had
received anterior treatment for one week. The conventional anterior
treatment was a standard burn therapy composition including 0.5 weight
percent of silver nitrate solution, mafenidate acetate, and 1 percent
silver sulfadiazine. All patients presented bacterial invasion
including streptococci and staphylococci. The tissue on all patients
had blisters and fibrinous exudate. Each patient applied 200 mg of
ointment containing 3 wt % tetrasilver tetroxide in 97 wt % petroleum
jelly three times daily to the arm burns, which were then covered by 3
layers of cotton roll bandages. The hands of each was covered by
gloves, which were changed every 10 days, that contained 500 mg of
ointment (3 wt % tetrasilver tetroxide and 97 wt % petroleum jelly).
The patients were evaluated every day for 30 days in the hospital,
without any outpatient treatment and with daily laboratory tests.

Group II: The other 7 patients were diagnosed as having third degree
burns, 40% to 65% BSA, large severity, on both hands, arms, feet, and
legs. All members received the same conventional anterior treatment as
the Group I patients.

Group IIA: Four of these patients had 40% BSA burns with lab results
showing no bacterial invasion. These patients had normal temperature
curves and hemograms, and they had skin surfaces that were pale and
anesthetic. These patients received electrolyte volume replacement.
They each applied 300 mg of the same ointment noted above 3 times per
day for 30 days, with hands in gloves containing 500 mg of the same
ointment and changed every ten days. Laboratory tests were performed
every week.

Group IIB: Three of the patients had 65% BSA burns with bacterial
invasion confirmed by lab testing. The temperature curve of each was
in the range of 39.5.degree. C. to 39.9.degree. C. at the beginning of
treatment. The hemograms of each showed a marked leucositosis, and the
skin surfaces were black, charred, and leathery. All patients in this
group had escharotomies before treatment and the fever did not
disappear. This group received the same treatment as Group IIA above.

Results

Group I: Over a period of 6 to 9 days, all patients observed the burn
lesions dry out, and all blisters and erythematous areas disappear.
Minimal fibrinous exudate was present, with no patient having a fever.
By day 14, the hemograms became normal. In 10 more days, i.e., by day
24, the lesions in the hands were evaluated. The skin was completely
healed and had no more fibrinous exudate. Skin cultures were normal
with no bacterial invasion reported. Skin necrosis did not develop in
any patient in this group, and all joint functions were preserved.
None of the patients had to be in a surgery room for wound cleaning,
removal, or debridement. Even after six months of conventional
treatment, the results do not match those after 30 days of treatment
using the compositions and methods of the present invention.

Group IIA: At day 10, the gloves were removed from all patients and
the skin of the hands was healed with no sign of skin necrosis. All
joint functions were preserved and no patient had wound bed
contractions. Escharotomies were not required, and none of the
patients developed sodium loss, hypokalemia, hypochloremia, alkalosis,
or methemoglobinuria, each of which is an adverse effect that may
occur in burn victims during or following conventional treatment. Over
a period of 13 to 20 days, all patients in this group had the injured
skin become normal in color and texture. No bacterial invasion was
present after the treatment and lab tests were normal.

Group IIB: At day 10, the gloves were removed from all patients and
the skin of the hands was healed about 50%. At day 20, the gloves were
removed again and the injured skin was all healed with no sign of skin
necrosis. All of the skin on the patients developed wound bed
contractions, but all joint functions were preserved. About one-third
of the patients developed sodium loss and hypokalemia, but none
developed hypochloremia, alkalosis, or methemoglobinuria. Over a
period of 23 to 28 days, the injured skin on all patients in this
group dried out and lab tests did not indicate the presence of
bacterial invasion. No fever was present and the black, charred,
leathery skin became normal skin. The patients of this group continued
with outpatient follow-up to evaluate the wound bed contractions,
examine the skin for cellulitis, and to consider excisional therapy.

Conclusions

The composition according to the invention used for treating second
degree burns, 20% BSA, of moderate severity with bacterial invasion
seemed to eliminate 100% of the bacteria and heal the injured skin in
a record time of 10 days. The use of gloves filled with a composition
prepared according to the invention to treat hand burns healed these
burns faster than any known conventional hand burn treatment. The
composition of the invention also prevented bacterial invasion of skin
burn injury having no bacterial invasion, prevented skin necrosis in
burn injuries, and increased the speed of healing of tissue caused by
thermal burn contact. With respect to third degree burns, the
composition of the invention caused sodium losses and hypokalemia, but
helped to preserve joint functions.

Example 3

Compositions of Invention Compared to Art

A comparison study was conducted of tetrasilver tetroxide, prepared in
accordance with the present invention, and a conventional monovalent
silver oxide. Benchmark Analytics of Center Valley Pa., a licensed
certifying laboratory, tested the efficacy of 2 ppm of each compound
against E. Coli. A 100,000 CFU/mL culture had a 43% kill, i.e., of
43,000 in 10 minutes contact time with the composition of the
invention, compared with a 37.3% reduction with the conventional
silver oxide of a 75,000 CFU/mL culture under the same conditions.
Since the tetrasilver tetroxide contains 87% silver by weight, and the
conventional silver oxide contains 93% silver by weight, the silver
does not appear to be primarily responsible for the antimicrobial
efficacy of the composition of the invention. Indeed, when one adjusts
the results to a content of 2 ppm silver, the tetrasilver tetroxide
kills 49,000, and the conventional compound kills only 30,000, each
calculated for a 100,000 CFU/mL culture, of E. Coli. Thus, the
antibacterial efficacy of the composition of the invention is believed
to facilitate the treatment or management of burns. In one embodiment,
the electron active compound can inhibit or prevent bacterial or other
pathogenic invasion of burns or skin grafts, which can facilitate
healing.

Based on all of the test data described above, the healing mechanism
associated with the use of the metal oxides of the invention to treat
and manage at least some skin diseases, without being bound by theory,
appears to involve mechanisms other than merely inhibiting or killing
pathogens and curing infections that tend to aggravate disease and
retard the natural healing process. The data indicate that healing is
brought about even in cases where no abnormal bacteria counts or
infection is evident. This suggests that the electron active
compound(s) may also act against auto-antibodies that trigger
autoimmune reactions associated with diseased tissue, as well as
against other non-pathogenic conditions or diseases, such as
circulatory or neurological conditions or diseases.

Although preferred embodiments of the invention have been described in
the foregoing Detailed Description, it will be understood that the
invention is not limited to the embodiments disclosed, but is capable
of numerous rearrangements and modifications of parts and elements
without departing from the spirit of the invention. It will be further
understood that the chemical and pharmaceutical details of the
compositions and methods of prevention, treatment, or management
herein may be slightly different or modified by one of ordinary skill
in the art without departing from the claimed invention.



United States Patent 6,645,531
Antelman November 11, 2003
Multivalent electron active compositions and methods of making and
using same

Abstract

The present invention is directed to pharmaceutical compositions that
include a therapeutically effective amount of at least one electron
active compound, or a pharmaceutically acceptable derivative thereof,
that has at least two polyvalent cations, at least one of which has a
first valence state and at least one of which has a second, different
valence state. Preferred compounds include Bi(III,V) oxide, Co(II,III)
oxide, Cu(I,III) oxide, Fe(II,III) oxide, Mn(II,III) oxide, and
Pr(III,IV) oxide, and optionally Ag(I,III) oxide. These compounds may
be in a crystalline state having metallic cations of two different
valences, or electronic states, in the inorganic crystal. In addition,
the invention relates to methods for prevention, management, or
treatment of a condition using these compounds or pharmaceutical
compositions including the same.
Inventors: Antelman; Marvin S. (Rehovot, IL)
Assignee: Marantech Holding LLC (East Province, RI)
Appl. No.: 09/692,126
Filed: October 20, 2000
Related U.S. Patent Documents

Application Number Filing Date Patent Number Issue Date
552172 Apr., 2000 6258385

Current U.S. Class: 424/635 ; 424/617; 424/630; 424/639; 424/646;
424/647; 424/648; 424/653
Current International Class: A61K 47/02 (20060101); A61K 8/02
(20060101); A61K 8/19 (20060101); A61Q 1/12 (20060101); A61K 33/24
(20060101); A61K 33/26 (20060101); A61K 33/32 (20060101); A61K 33/34
(20060101); A61K 33/38 (20060101); A61K 9/06 (20060101); A61Q 19/00
(20060101); A61K 033/34 (); A61K 033/24 (); A61K 033/26 (); A61K
033/32 ()
Field of Search: 424/600,617,618,630,634,639,646,647,648,653
References Cited [Referenced By]
U.S. Patent Documents

3923982 December 1975 Lamand et al.
4447254 May 1984 Hughes et al.
4828832 May 1989 De Cuellar et al.
4952411 August 1990 Fox, Jr. et al.
5017295 May 1991 Antelman
5073382 December 1991 Antelman
5078902 January 1992 Antelman
5089275 February 1992 Antelman
5098582 March 1992 Antelman
5211855 May 1993 Antelman
5223149 June 1993 Antelman
5334588 August 1994 Fox, Jr. et al.
5336416 August 1994 Antelman
5336499 August 1994 Antelman
5571520 November 1996 Antelman
5612019 March 1997 Gordon et al.
5676977 October 1997 Antelman
5772896 June 1998 Denkewicz, Jr. et al.
Foreign Patent Documents

2000060976 Feb., 2000 JP

Other References

STN/CAS online, file CAPLUS, Acc. No. 1999:748588, Doc. No. 131:340719
(JP 11322408 A2 (Ohne et al.), Nov. 24, 1999), Abstract.* .
STN/CAS online, file CAPLUS, Acc. No. 1997:195107, Doc. No. 126:182656
(JP 09012415 A2 (Doi et al.), Jan. 14, 1997), Abstract.* .
Antelman, Marvin S.; "Silver (II,III) Disinfectants"; Soap/Cosmetics/
Chemical Specialties, Mar. 1994, pp. 52-59. .
Antelman, Marvin S.; Abstracts of American Chemical Society;
1992(203). .
Antelman, Marvin S.; "Anti-Pathogenic Multivalent Silver Molecular
Semiconductors"; Precious Metals; 1992(16); pp. 141-149. .
Antelman, Marvin S.; "Multivalent Silver Bactericides"; Precious
Metals; 1992(16); pp. 151-163. .
Fung, Man C. and Bowen, Debra L.; "Silver Products for Medical
Indications: Risk-Benefit Assessment", Clinical Toxicology, 1996, pp.
119-126. .
Dorland et al., Dorland's Illustrated Medical Dictionary,
Philadelphia: W.B. Saunders Company, 1994, 28.sup.th Edition, p. 351,
759, and 760. .
Gennaro, A., Remington's Pharmaceutical Sciences, Easton, PA: Mack
Publishing Company, 1985, 17.sup.th Edition, p. 1573-1575, 1585-1594,
and 1601..

Primary Examiner: Dees; Jose' G.
Assistant Examiner: Choi; Frank
Parent Case Text


CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of Application No.
09/552,172, filed Apr. 18, 2000, now U.S. Pat. No. 6,258,385 and
claims benefit of Provisional Application No. 60/174,793, filed Jan.
6, 2000, No. 60/184,053, filed Feb. 22, 2000, and No. 60/214,503,
filed Jun. 28, 2000.
Claims


What is claimed is:

1. A method of halting, diminishing, or inhibiting the growth of at
least one of a bacterium, a fungus; a parasitic microbe, and a virus,
which method comprises administering to a human being a
therapeutically effective amount of at least one electron active
compound that has at least two polyvalent cations, at least one of
which has a first valence state and at least one of which has a second
different valence state, wherein the at least one electron active
compound comprises a metal oxide selected from the group consisting of
Bi(III,V) oxide, Co(II,III) oxide, Cu(I,III) oxide, Mn(II,III) oxide,
Pr(III,IV) oxide, Tb.sub.4 O.sub.7, or a mixture thereof.

2. Tetracopper tetroxide, which comprises two copper(I) ions, two
copper(III), ions and four oxygen atoms in a crystal lattice.

3. A process for preparing tetracopper tetroxide, having two copper(I)
ions, two copper(III) ions, and four oxygen atoms in a crystal
lattice, which process comprises: combining a copper(I)-containing
compound and a caustic solution to form a reactant solution; and
heating the reactant solution to a temperature and for a time
sufficient to produce a detectable amount of a tetracopper tetroxide.

4. The process of claim 3, wherein the copper(I)-containing compound
comprises a non-solvated inorganic copper(I) oxide.

5. The process of claim 4, wherein non-solvated inorganic copper(I)
oxide comprises cuprous oxide.

6. The process of claim 3, wherein the caustic solution comprises a
strong caustic base and a peroxy acid salt.

7. The process of claim 3, wherein the strong caustic base comprises a
hydroxide salt, and wherein the peroxy acid salt comprises a
persulfate.
Description


FIELD OF THE INVENTION

The present invention relates to electron active compounds and
compositions that have polyvalent cations in their crystal lattices.
In addition, the present invention also includes a method of making
such electron active compounds. The present invention also relates to
methods for the prevention, treatment, or management of conditions, or
symptoms thereof, by administering one or more such compounds or
compositions.

BACKGROUND OF THE INVENTION

Tetrasilver tetroxide has been demonstrated to possess unique
properties arising from electrostatic concepts of metal cation
interaction. Such silver molecules have also been disclosed for
various uses, as they are reported to be non-toxic to animals and
humans. M. Antelman, "Anti-Pathogenic Multivalent Silver Molecular
Semiconductors," Precious Metals, vol. 16:141-149 (1992); M. Antelman,
"Multivalent Silver Bactericides," Precious Metals, vol. 16:151-163
(1992). For example, tetrasilver tetroxide activated with an oxidizing
agent is disclosed for use in bactericidal, fungicidal, and algicidal
use, such as in municipal and industrial water treatment applications
and for the treatment of AIDS.

A variety of sources also report the use of certain divalent silver
compounds for water treatment, as well as the use of such compounds,
typically in combination with certain oxidizing agents, metals, or
other compounds, as disinfectants, bactericides, algicides, and
fungicides. One source also reports a single in vitro study of the use
of such compounds for the treatment of AIDS. These sources include M.
Antelman, "Silver (II, III) Disinfectants," Soap/Cosmetics/Chemical
Specialties, pp. 52-59 (Mar., 1994), and U.S. Pat. Nos. 5,017,295;
5,073,382; 5,078,902; 5,089,275; 5,098,582; 5,211,855; 5,223,149;
5,336,416; and 5,772,896.

U.S. Patent No. 5,336,499 discloses tetrasilver tetroxide and
persulfate compositions having certain in vitro anti-pathogenic
properties, i.e., bactericidal, fungicidal, viricidal, and algicidal,
in certain concentrations as low as 0.3 ppm, particularly in nutrient
broth cultures. The persulfate is disclosed as being an oxidizing
agent that activates the tetroxide crystals. Also disclosed are an in
vitro study regarding the inhibition of yeast growth in nutrient broth
and the formulation of a gynecological cream and douche based on these
results, and a report of an in vitro AIDS test with the compositions
indicating total suppression of the virus at 18 ppm.

U.S. Pat. No. 5,571,520 discloses the use of molecular crystals of
tetrasilver tetroxide, particularly with oxidizing agents to enhance
the efficiency of such devices, for killing pathogenic microorganisms,
such as staph infections. Amounts of 10 ppm sodium persulfate as an
oxidizing agent were used with certain amounts of silver tetroxide in
the reported in vitro testing. One human study involved in vivo curing
of a gynecological yeast infection with 10 ppm of the silver tetroxide
and 40 ppm sodium persulfate. Other in vivo topical studies report in
conclusory fashion the cure of a single case of athlete's foot with a
solution of 100 ppm of the composition and the cure of a single case
of toenail fungus with a 25% suspension of the composition.

U.S. Pat. No. 5,676,977 discloses intraveneously injected tetrasilver
tetroxide crystals used for destroying the AIDS virus, AIDS
synergistic pathogens, and immunity suppressing moieties (ISM) in
humans. The crystals were formulated for a single injection at about
40 ppm of human blood. This reference also discloses the compositions
cause hepatomegaly, also known as enlarged liver, albeit with no
reported loss of liver function.

The aforementioned references report detailed descriptions of the
mechanism via which the multivalent silver molecular crystal devices
were believed to operate. A discussion of such results and concepts
was presented at a Seminar entitled "Incurable Diseases
Update" (Weizmann Institute of Science, Rehovot, Israel, Feb. 11,
1998). The title of this presentation was "Beyond Antibiotics, Non
Toxic Disinfectants and Tetrasil.TM. (a composition including
tetrasilver tetroxide)." In this paper, it was reported that the
effects of the electron transfer involved with respect to the
tetroxide, rendered it a more powerful germicide than other silver
entities. Other patents cover multivalent silver antimicrobial
compositions, e.g., U.S. Pat. No. 5,017,295 for Ag(II) and U.S. Pat.
No. 5,223,149 for Ag (III). These are stronger antimicrobial agents
than Ag (I) compounds, but they pale by comparison to tetrasilver
tetroxide. Likewise, colloidal silver that derives its germicidal
properties from trace silver (I) ions it generates in various
environments is also less effective. Accordingly, the oligodynamic
properties of these entities may be summarized as follows, which is
referred to as the Horsfal series:

Another property of the tetrasilver tetroxide is that it does not
stain organic matter such as skin in like manner as Ag(I) compounds
do. In addition, it is light stable.

Further, synthetic routes for making Bi(III,V) oxide are detailed and
reviewed in Gmelins Handbuch DerAnorganischen Chemie, vol. 16:642
(1964). Also, Co(II,III) oxide, Fe(II,III) oxide, Mn(II,III) oxide,
and Pr(III,IV) oxide can all be found in nature. These five
multivalent metal oxides are also all available commercially.

In view of the beneficial properties of tetrasilver tetroxide, it
could be desirable to find other medicinal uses for this compound, as
well as to discover other electron active metal oxides that provide
similar properties.

SUMMARY OF THE INVENTION

The present invention relates to pharmaceutical compositions that
include a therapeutically effective amount of at least one electron
active compound, or a pharmaceutically acceptable derivative thereof,
that has at least two polyvalent cations, at least one of which has a
first valence state and at least one of which has a second, different
valence state. Advantageously, the pharmaceutical composition may have
antipathogenic efficacy. Preferably, the at least one electron active
compound includes a metal oxide. In one embodiment, the metal oxide
includes at least one of bismuth, cobalt, copper, iron, manganese,
praseodymium, or a combination thereof. Preferably, in that
embodiment, the metal oxide includes at least one of Bi(III,V) oxide,
Co(II,III) oxide, Cu(I,III) oxide, Fe(II,III) oxide, Mn(II,III) oxide,
Pr(III,IV) oxide, or a combination thereof. In another embodiment, the
metal oxide can include Ag(I,III) oxide. Alternately, the
pharmaceutical composition does not include tetrasilver tetroxide. In
another alternate embodiment, the pharmaceutical composition does not
include tricobalt tetroxide. In one embodiment, the pharmaceutical
composition may include at least two different electron active
compounds. In another embodiment, the compound may be in powder or
granular form.

In a preferred embodiment, the first valence and the second valence of
the at least two polyvalent cations differ by at least 1, preferably
by 1 or 2. In another preferred embodiment, the first valence and the
second valence of the at least two polyvalent cations differ by more
than 2. Advantageously, the electron active compound has at least one
polyvalent cation which has an EMF.sup.ox of at least about +0.1
Volts.

In one embodiment, the amount of the at least one electron active
compound is present in an amount from about1 ppm to 500,000 ppm, based
on the weight of the composition. If desired, the pharmaceutical
composition can include a pharmaceutically acceptable carrier.
Optionally, the composition can also include an oxidizing agent,
preferably present in an amount sufficient to enhance the efficacy of
the active compound but insufficient to cause skin irritation.
Preferably, the oxidizing agent includes a peroxy acid salt of a
persulfate.

In a preferred embodiment, the at least one compound has antimicrobial
efficacy, preferably of at least about 20%. In another embodiment, the
antimicrobial efficacy is at least about 50%. In yet another
embodiment, the antimicrobial efficacy is at least about 80%. In these
embodiments, about 100 ppm of the at least one compound is placed in
contact for about 10 minutes with microbes having a cell density of
approximately 75,000 CFU/mL.

Also an aspect of the present invention is a pharmaceutical
composition comprising tetracopper tetroxide compound. Advantageously,
the tetracopper tetroxide contains two copper(I) ions, two copper(III)
ions, and four oxygen atoms in a crystal lattice.

Another aspect of the present invention is a method of preventing,
treating, or managing a condition of a patient which includes
administering a therapeutically effective amount of at least one of
the electron active compounds described herein, or a pharmaceutically
acceptable derivative thereof, to prevent, treat, or manage the
condition, or a symptom thereof. In one embodiment, the method
excludes tetrasilver tetroxide. In a preferred embodiment, the patient
is a mammal, preferably, a human. Advantageously, the electron active
compound(s) can be administered topically, parenterally, or
transdermally, preferably in an amount from about 5 ppm to 500,000
ppm, based on the weight of the composition. In one embodiment, at
least two different electron active compounds are administered.

In another embodiment, the method can include administering one or
more additional different therapeutic agents, present in an amount
sufficient to facilitate the prevention, treatment, or management of
the condition. In this embodiment, the one or ore additional
therapeutic agents may optionally be administered concurrently with
the electron active compound(s).

Another aspect of the present invention relates to a method of
facilitating the killing of a pathogen which includes administering a
therapeutically effective amount of at least one electron active
compound, or a pharmaceutically acceptable derivative thereof, that
has at least two polyvalent cations, at least one of which having a
first valence state and at least one of which having a second
different valence state.

The present invention also involves a method of inhibiting the growth
of a pathogen which comprises administering a therapeutically
effective amount of at least one electron active compound, or a
pharmnaceutically acceptable derivative thereof, that has at least two
polyvalent cations, at least one of which having a first valence state
and at least one of which having a second different valence state.

In one embodiment, these methods exclude the administration of
tetrasilver tetroxide. In either of these methods, the pathogen can
include a gram-positive bacillus or coccus; a gram-negative bacillus
or coccus; an acid-fast bacterium; another type of bacterium; a
fungus; a parasitic microbe; a virus; or a combination thereof.

Tetracopper tetroxide, containing two copper(I) ions, two copper(III)
ions, and four oxygen atoms, is one preferred electron active
compound, while the administration of tetrasilver tetroxide for
treating certain conditions is excluded.

In addition, the present invention relates to a process for preparing
tetracopper tetroxide, which includes: combining a copper(I)-
containing compound and a caustic solution to form a reactant
solution; and heating the reactant solution to a temperature and for a
time sufficient to produce a detectable amount of the tetracopper
tetroxide compound. Advantageously, the copper(I)-containing compound
includes a non-solvated inorganic copper(I) oxide, such as cuprous
oxide.

The caustic solution generally contains a strong caustic base and a
peroxy acid salt. Preferably, the strong caustic base includes a
hydroxide salt, and the peroxy acid salt includes a persulfate.

Another aspect of the invention relates to method water with the
compounds or compositions of the present invention.

DEFINITIONS

Some of the terms used in connection with the invention can be defined
as follows:

The term "condition," as used herein, should be understood to refer to
a traditionally identified disease, as well as a disorder, an
affliction, or an ailment, particularly including those noted herein.

The terms "prevent," "preventing," and "prevention," as used herein,
refer to stopping or hindering a condition, symptom, or pathogen
causing a condition, in a patient who is at risk of suffering from
such a condition. This also includes reducing the frequency or
severity, or both, of the occurrence of such conditions or one or more
symptoms thereof.

The terms "manage," "managing," and "management," as used herein,
includes controlling those conditions which cannot be cured
completely, reducing the time of affliction of such conditions, and
the like. Preferably, the compositions prevent, treat, or manage such
conditions without superficially discoloring the skin, i.e., no
discoloration to the naked eye. In one embodiment, the invention
relates to the treatment or management, while in another embodiment
the invention relates to the prevention, of the diseases or conditions
disclosed and claimed herein. The terms also include the use of the
compounds or compositions of the invention to facilitate the halting,
diminishing, or inhibiting of the growth or proliferation of pathogens
that may accentuate, amplify, exacerbate, or cause, either directly or
indirectly, a condition and/or a symptom thereof.

The term "patient" as used herein refers to animals, particularly to
mammals. In one preferred embodiment, the term patient refers to
humans.

The terms "adverse effects," "adverse side effects," and "side
effects," as used herein, include, but are not limited to, cardiac
arrhythmia, cardiac conduction disturbances, appetite stimulation,
weight gain, sedation, gastrointestinal distress, headache, dry mouth,
constipation, diarrhea, drug-drug interactions, superficial
discoloration of the skin, dry skin, hepatomegaly, fever, fatigue, and
the like. The term "cardiac arrhythmia" includes, but is not limited
to, ventricular tachyrhythmia, torsades de pointes, Q.sub.T
prolongation, and ventricular fibrillation.

The phrase "therapeutically effective amount" when used herein in
connection with the compositions and methods of the invention, means
that amount of electron active metal oxide compound(s) or
composition(s), or a derivative thereof, which, alone or in
combination with other drugs, provides a therapeutic benefit in the
prevention, treatment, or management, of a condition. In one
embodiment, the effective amount is one or more metal oxide compounds
or compositions as the sole active ingredient. Different
therapeutically effective amounts may be applicable for each
condition, as will be readily known or determined by those of ordinary
skill in the art.

The term "substantially free" means less than about 10 weight percent,
preferably less than about 5 weight percent, more preferably less than
about 1 weight percent, and most preferably less than about 0.1 weight
percent. For example, a composition may be substantially free of added
oxidizing agent or of added persulfate according to the invention.

The term "about," as used herein, should generally be understood to
refer to both numbers in a range of numerals. Moreover, all numerical
ranges herein should be understood to include each whole integer
within the range.

The term "substantial," as used herein, means at least about 75%,
preferably at least about 90%, more preferably at least about 95%,
most preferably at least about 99%.

The term "valence state," as used herein, should be understood to
refer to the charge on a given ion or to the charge that may be
assigned to a given ion based on its electronic state.

The terms "inhibit," "inhibiting," or "inhibits," as used herein when
referring to growth of an item, should be understood to refer to the
act of stopping that growth, whether permanently or temporarily, or of
reducing the rate of that growth, either permanently or temporarily.

DETAILED DESCRIPTION OF THE INVENTION

The tetrasilver tetroxide compounds mentioned in the background are
one type of electron active compound having multivalent cations in its
crystal lattice. Various additional electron active compounds have now
also been identified, as well as methods for making and using the same
for treating various pathogenic and non-pathogenic conditions or
disorders. The electron active compounds of the present invention are
believed to have unique crystal structures in that, in the case of the
metal oxides, there are generally atoms of the same element in the
crystal that have at least two different valences, typically at least
one lower-valent metal cation and at least one higher-valent metal
cation, for example, such as Co(II) and Co(III), respectively.
Exemplary electron active metal oxide compounds according to the
invention include, but are not limited to, Ag(I,III), Co(II,III),
Pr(III,IV), Bi(III,V), Fe(II,III), Mn(II,III), and Cu(I,III) oxides.
In another embodiment, Tb(III,IV) oxide, Tb.sub.4 O.sub.7, or
tetraterbium heptoxide, is one electron active metal oxide compound
according to the invention. As discussed below, pharmaceutical
compositions including one or more of such oxide compounds are useful
for treating various conditions. The composition of such exemplary
electron active metal oxides is shown in tabular form below:

Lower-valent Higher-valent e Formula Metal cations ion # ion # 2
Ag.sub.4 O.sub.4 Ag(I,III) Ag.sup.+ 2 Ag.sup.+3 2 1 Co.sub.3 O.sub.4
Co(II,III) Co.sup.+2 1 Co.sup.+3 2 2 Pr.sub.6 O.sub.11 Pr(III,IV)
Pr.sup.+3 2 Pr.sup.+4 4 2 Bi.sub.2 O.sub.4 Bi(III,V) Bi.sup.+3 1
Bi.sup.+5 1 1 Fe.sub.3 O.sub.4 Fe(II,III) Fe.sup.+2 1 Fe.sup.+3 2 1
Mn.sub.3 O.sub.4 Mn(II,III) Mn.sup.+2 1 Mn.sup.+3 2 2 Cu.sub.4 O.sub.4
Cu(I,III) Cu.sup.+ 2 Cu.sup.+3 2 e - total number of electrons
believed to be exchanged; # - number of particular ion type per
formula unit.

Without being bound to theory, it is believed that the electron active
compounds operate against pathogens by transferring electrons between
their lower-valent ions and their higher-valent ions in the crystal,
thereby contributing to the death of pathogens by traversing their
cell membrane surface. It would seem that this, in effect,
"electrocutes" the pathogens. While these compounds have also been
discovered to be suitable for use in the prevention, treatment, and
management of other non-pathogenic conditions and disorders, such as
autoimmune disorders, circulatory disorders, neurological disorders,
and the like, the mechanism by which such conditions or disorders are
prevented, treated, or managed has not yet been fully understood. In
any event, the electrons in proximity to pathogens are believed to be
perturbed from their balanced crystals by such labile groups as NH,
NH.sub.2, S--S, and SH, which can be present, for example, in a
pathogen cell membrane. It is believed, however, that normal cells
will not be significantly affected because they do not proliferate
rapidly enough to expose these labile bonds sufficiently for the bonds
to be substantially affected.

The crystals in the electron active compounds are not believed to be
disturbed unless more stable complexes are formed with ligands, for
example, such as those comprising a pathogen cell membrane surface in
a dynamic state. Indeed, the end result of electron transfer, which is
a redox reaction, results in the lower-valent metal ions being
oxidized to one valence state higher and the higher-valent metal ions
being reduced to one valence state lower. In one embodiment, the
oxidation of the lower-valent metal ions and the reduction of the
higher-valent metal ions both result in ions having the same oxidation
state. Examples of such an embodiment occur when the valence
difference between the metal ions in the electron active molecular
crystal is 2 and such examples include, but are not limited to,
Ag(I,III), Bi(III,V), and Cu(I,III) oxides. In another embodiment, the
oxidation of the lower-valent metal ions and the reduction of the
higher-valent metal ions result in ions having opposite oxidation
states (e.g., ions with a +2 valence state are oxidized to +3, while
the ions with a +3 valence state are reduced to +2). Examples of such
an embodiment occur when the valence difference between the metal ions
in the electron active molecular crystal is 1 and such examples
include, but are not limited to, Co(II,III), Fe(II,III), Mn(II,III),
and Pr(III,IV) oxides.

The metal ion of certain electron active compounds may exhibit a
distinct affinity for certain elements of ligands, for example, such
as sulfur, oxygen, or nitrogen, particularly when present in a
pathogen's cell membrane. In many cases, the metal ion will not merely
bind to these elements, but will actually form chelate complexes with
their ligands. The classic example of this is Ag(I,III) oxide, the
monovalent silver ion of which has an affinity for sulfur and nitrogen
and the oxidized/reduced divalent ion of which forms chelate complexes
with, for example, mercapto or amino groups. Thus, the electron active
compound attraction for the cell membrane surfaces, for example, of
pathogens, is believed to be driven by powerful electrostatic forces.

Without being bound by theory, the electron exchange may be depicted,
for example, by the following series of redox half reactions:

metal(III,IV) metal(III,V) metal(I,III) oxides metal(II,III) oxides
oxides oxides Ag.sup.+ - e = Ag.sup.+2 Co.sup.+2 - e = Co.sup.+3
Pr.sup.+3 - e = Pr.sup.+4 Bi.sup.+3 - e = Bi.sup.+4 Ag.sup.+3 + e =
Ag.sup.+2 Co.sup.+3 + e = Co.sup.+2 Pr.sup.+4 + e = Pr.sup.+3 Bi.sup.
+5 + e = Bi.sup.+4 Cu.sup.+ - e = Cu.sup.+2 Fe.sup.+2 - e = Fe.sup.+3
Cu.sup.+3 + e = Cu.sup.+2 Fe.sup.+3 + e = Fe.sup.+2 Mn.sup.+2 - e =
Mn.sup.+3 Mn.sup.+3 + e = Mn.sup.+2

For each redox reaction, there is believed to be an electromotive
force, which is the voltage potential of the oxidizing the higher-
valent ion in the metal oxide crystal. This is denoted herein as
EMF.sup.OX. In addition to the electromotive force of oxidation, there
is believed to be an associated reduction reaction involving the lower-
valent ion in the metal oxide crystal. This reduction reaction may be
represented simply, as tabulated above, or may represent the
interaction with, for example, a ligand present on a pathogen cell
membrane surface, such as one containing sulfur or nitrogen.
Associated with the reduction reaction is another electromotive force,
or voltage potential of the reducing the lower-valent ion. This is
denoted herein as EMF.sup.RE.

When the metal ions of the electron active metal oxide interact with,
for example, a sulfur-containing ligand, the affinity of the metal ion
for sulfur affects EMF.sup.RE. The stability of a particular metal
sulfide is an approximation of the affinity of a metal ion for sulfur.
The following approximate association constants for sulfides indicate
the trend in relative affinity of each metal ion for sulfur:

Ag(I) 49 Cu(I) 47 Co(II) 26 Fe(II) 19 Mn(II) 15

In general, the more stable the compound, the more negative its
reduction potential in the reduction reaction, for example, in the
case of elemental silver:

In the case of tetrasilver tetroxide, there is a reduction reaction
where Ag(I) is oxidized and an oxidation reaction where Ag(III) is
reduced, as follows: ##EQU1##

The voltage that is discharged from a redox reaction of the electron
active metal oxides of the present invention, which voltage is denoted
herein as the "electrocution voltage," is the combination of the
oxidizing cation's reduction potentials and the reducing cation's
reduction potential (i.e., EMF.sup.OX -EMF.sup.RE) In the case of
tetrasilver tetroxide, the "electrocution voltage" is 2.92 volts. The
oxidizing cation's reduction potentials, EMF.sup.OX, of exemplary
metal oxides according to the present invention are tabulated below:

Formula Metal cations EMF.sup.ox Ag.sub.4 O.sub.4 Ag(I,III) 2.02
Co.sub.3 O.sub.4 Co(II,III) 1.81 Pr.sub.6 O.sub.11 Pr(III,IV) 2.86
Bi.sub.2 O.sub.4 Bi(III,V) 1.59 Fe.sub.3 O.sub.4 Fe(II,III) 0.77
Mn.sub.3 O.sub.4 Mn(II,III) 1.54 Cu.sub.4 O.sub.4 Cu(I,III) 1.80

As noted from the above table, praseodymium-, cobalt-, and copper-
based oxides are believed to be stronger antipathogenic agents or to
form better pharmaceutical compositions than manganese-, bismuth-, and
iron-based oxides, and in one embodiment they are preferred for this
reason. Nevertheless, in certain cases, iron exhibits stronger
antipathogenic characteristics, particularly antimicrobial
characteristics, compared to manganese.

Another factor, however, particularly in antipathogenic or
antimicrobial efficacy, can be the sulfur/nitrogen composition, for
example, of cell membranes. For example, Staphylococcus aureus
bacteria, in a culture having a cell density of 30,000 CFU/mL, exhibit
significant mortality from exposure to 100 ppm of Bi(III,V) oxide for
about 10 minutes, but no significant mortality from exposure to the
same concentrations of Fe(II,III) and Mn(II,III) oxides for the same
contact time. This result might be explained by the far greater
stability of bismuth(III) sulfide, and thus the far greater affinity
of bismuth(III) for sulfur, than either of the iron(II) or
manganese(II) analogs.

The electron active metal oxide compounds and compositions of the
present invention may be used in any form which sufficiently retains
their antipathogenic character, or other non-pathogenic ability, to
prevent, treat, or manage one or more of the conditions noted herein.

These compounds or compositions may be used as antipathogenic agents,
such as antimicrobial, antibacterial, antiviral, or anti-algal agents,
or a combination thereof. In another embodiment, the compounds or
compositions may be used for preventing, treating, and/or managing
various conditions that are non-pathogenic. For example, non-
pathogenic conditions are believed to include certain autoimmune
disorders, neurological disorders, and circulatory disorders. While
the exact mechanism of the activity of such compounds or compositions
is not described herein, nonetheless, suitable prevention, treatment,
and/or management of such non-pathogenic conditions may be obtained by
administering the compounds or compositions of the invention as
described herein and as will be readily apparent to one of ordinary
skill in the art.

The compositions and methods of the invention advantageously prevent,
treat, or manage dermatological diseases or conditions. The conditions
against which the electron active compounds, such as metal oxides, of
the present invention have utility include, but are not limited to,
Madura foot, actinomycosis, oral actinomycosis, anthrax, food
poisoning, botulism, wound infections, pseudomembranous colitis,
colitis, gas gangrene, gangrene, tetanus, diphtheria, pharyngeal
diphtheria, pleomorphic laryngeal diphtheria, cutaneous diphtheria,
endocarditis, bacteremia, urinary tract infections, listerosis,
meningitis, miscarriage, narcodiosis, acne, skin lesions, abscesses,
toxic shock syndrome, prosthesis contamination, dental caries, plaque,
gum disease, gingivitis, subacute endocarditis, bacterial pneumonia,
otitis, sinusitis, cat scratch fever, septicemia, abdominal and pelvic
abscesses, Oroya fever, systemic Oroya fever, verruga peruana,
cutaneous verruga peruana, whooping cough, Lyme disease, epidemic
relapsing fever, brucellosis, granuloma inguinale granulomatic,
donovanosis, gastroenteritis, nosocomial infections, tularemia,
bacterial vaginitis, urethritis, bacterial conjunctivitis, chancroid,
otitis media, chronic gastritis, peptic ulcer, diarrhea, Legionnaires'
disease, leptospirosis, gonorrhea, arthritis, periodontal disease,
salmonellosis, typhoid fever, shigellosis, rat bite fever,
pharyngitis, scarlet fever, syphilis, cholera, Asiatic cholera,
Yersina arthritis, bubonic plague, chronic pulmonary disease, Hansen's
disease, leprosy, tuberculosis, dermal tuberculosis, psittachosis,
ornithosis, conjunctivitis, trachoma, lymphogranuloma venereum,
genital tract infections, Q fever, primary atypical pneumonia,
rickettsial pox, typhus, epidemic typhus, Rocky Mountain spotted
fever, tsutsugamushi fever, nongonococcal urethritis, human
erlichiosis, meningococcal meningitis, skin infections, corneal
infections, external ear infections, candidiasis, monoiliasis, thrush,
candidosis, mucositis, bacteremia, hepatitis, hepatitis A, hepatitis
B, hepatitis C, hepatitis E, coccidiomycosis, lymphadenitis,
balantidiasis cryptosporidosis, amoebiasis, amoebic dysentery,
giardiasis, giardia enteritis, leishmaniasis, Kala-azar, malaria,
toxoplasmosis, trypanosomiasis, Chagas disease, African sleeping
sickness, dengue, Japanese encephalitis, Rift Valley fever, Ebola
hemorrhagic fever, Venezuelan hemorrhagic fever, hantavirus pulmonary
syndrome, hemorrhagic fever with renal syndrome, cytomegalovirus
infection, poliomyelitis, West Nile virus disease, influenza, measles,
condyloma, encephalitis, ankylosing spondylitis, arteritis,
inflammatory bowel disease, polyarteritis nodosa, rheumatic fever,
systemic Lupus erythematosus, Alzheimer's disease, multiple sclerosis,
osteoporosis, Crohn's disease, strep throat, yellow fever, eczema,
psoriasis, dernatitis, disease-induced skin ulcers, undefined tropical
diseases, shingles, rashes, heat rashes, bedsores, cold sores,
blisters, boils, herpes simplex, acne, pimples, skin chafing, skin
cracking, itchiness, skin peeling, warts, one or more symptoms
thereof, or any combination thereof. In another embodiment, the
condition includes HIV (AIDS), or one or more symptoms. It should be
understood that the invention includes the use of the compounds or
compositions to prevent, treat, or manage each of these conditions
individually or multiple conditions concurrently or sequentially.
Thus, the prevention, treatment, or management of each condition
should be understood as a separate embodiment.

The pathogens which may be killed by, or the growth or proliferation
of which may be halted, diminished, or inhibited by, the electron
active metal oxides of the present invention include, but are not
limited to, gram-positive bacilli and cocci; gram-negative bacilli and
cocci; acid-fast bacteria; other bacteria; fungi; parasitic microbes,
e.g., protozoa; and viruses.

Examples of gram-positive bacilli and cocci include, but are not
limited to, Actinomedurae, Actinomyces israelii, Bacillus anthracis,
Bacillus cereus, Clostridium botulinum, Clostridium difficile,
Clostridium perfringens, Clostridium tetani, Corynebacterium,
Enterococcusfaecalis, Listeria monocytogenes, Nocardia,
Propionibacterium acnes, Staphylococcus aureus, Staphylococcus
epiderm, Streptococcus mutans, Streptococcus pneumoniae, and
combinations thereof.

Examples of gram-negative bacilli and cocci include, but are not
limited to, Afipia felis, Bacteriodes, Bartonella bacilliformis,
Bortadella pertussis, Borrelia burgdorferi, Borrelia recurrentis,
Brucella, Calymmatobacterium granulomatis, Campylobacter, Escherichia
coli, Francisella tularensis, Gardnerella vaginalis, Haemophilius
aegyptius, Haemophilius ducreyi, Haemophilius influenziae, Heliobacter
pylori, Legionella pneumophila, Leptospira interrogans, Neisseria
meningitidia, Porphyromonas gingivalis, Providencia sturti,
Pseudomonas aeruginosa, Salmonella enteridis, Salmonella typhi,
Serratia marcescens, Shigella boydii, Streptobacillus moniliformis,
Streptococcus pyogenes, Treponema pallidum, Vibrio cholerae, Yersinia
enterocolitica, Yersinia pestis, and combinations thereof.

Examples of acid-fast bacteria include, but are not limited to,
Myobacterium avium, Myobacterium leprae, Myobacterium tuberculosis,
and combinations thereof.

Examples of other bacteria not falling into the other three categories
include, but are not limited to, Bartonella henseiae, Chlamydia
psittaci, Chlamydia trachomatis, Coxiella bumetii, Mycoplasma
pneumoniae, Rickettsia akari, Rickettsia prowazekii, Rickettsia
rickettsii, Rickettsia tsutsugamushi, Rickettsia typhi, Ureaplasma
urealyticum, Diplococcus pneumoniae, Ehrlichia chafensis,
Enterococcusfaecium, Meningococci, and combinations thereof.

Examples of fungi include, but are not limited to, Aspergilli,
Candidae, Candida albicans, Coccidioides immitis, Cryptococci, and
combinations thereof.

Examples of parasitic microbes include, but are not limited to,
Balantidium coli, Cryptosporidium parvum, Cyclospora cayatanensis,
Encephalitozoa, Entamoeba histolytica, Enterocytozoon bieneusi,
Giardia lamblia, Leishmaniae, Plasmodii, Toxoplasma gondii,
Trypanosomae, trapezoidal amoeba, and combinations thereof.

Examples of viruses include, but are not limited to, Arboviruses,
Ebola virus, Guanarito virus, Hanta virus, Hantaan virus, Hepatitis A,
Hepatitis B, Hepatitis C, Hepatitis E, other Hepatitis viruses, Herpes-
type viruses, Poliovirus, West Nile virus, Echo virus, and
combinations thereof.

The antipathogenic or non-pathogenic compositions of the present
invention may optionally further include the use of one or more
additional therapeutic agents known to treat a condition, or a symptom
thereof. Examples of such additional therapeutic agents include, but
are not limited to, chelating agents, vitamins, minerals, silica
hydride microclusters, analgesics, Sambucol.TM., aspirin, and the
like.

The electron active metal oxide compounds of the present invention may
also be used for water treatment, for example, as disclosed in U.S.
Pat. No. 5,223,149 and 5,336,416. Optionally but preferably, the
electron active metal oxides used for treating a body of water are any
listed above, more preferably provided that the metal oxide does not
include tetrasilver tetroxide. It is also more preferable, in the
previous embodiment, that the metal oxide does not include tetracopper
tetroxide.

The administration of one or more active ingredients and/or optional
therapeutic agent(s), in accordance with the methods of the invention
may occur together, concurrently but separately, sequentially, or a
combination thereof. The optional additional therapeutic agent is
generally a compound other than an electron active metal oxide
compound.

The antipathogenic or antimicrobial performance of certain metal
oxides may be improved or enhanced by the presence of an oxidizing
agent. This is particularly the case when the metal oxide compounds or
compositions are present in low amounts, i.e., typically less than 45
ppm, and more commonly when present in an amount less than about 40
ppm, based on the weight of the composition. In such situations, an
oxidizing agent may be included in certain compositions of the
invention in small amounts when the compositions are administered by
certain routes. In such an embodiment, the oxidizing agent includes a
peroxy acid salt, preferably a Group I salt of a persulfate, more
preferably potassium persulfate. In another embodiment, the oxidizing
agent includes the same peroxy acid salt which was present as a
starting material in the reaction to form the particular electron
active metal oxide. The oxidizing agent may advantageously be present
in the composition in amounts from about 1 ppm to 500 ppm, based on
the weight of the composition. In alternate embodiments, there may be
from about 5 ppm to 200 ppm or from about 10 ppm to 100 ppm of
oxidizing agent, based on the weight of the composition.

It is believed that the additional presence of certain types or
amounts of oxidizing agent(s) may tend to irritate the skin,
particularly when the compound or composition including metal oxide(s)
is present in large amounts, such as greater than 50 ppm, based on the
weight of the composition. In one embodiment, as more compound or
composition is administered, a correspondingly smaller amount of
undesirable oxidizing agent is required. Thus, in some embodiments, it
has been found that the additional oxidizing agent is unnecessary and
in fact undesirable for the purpose of treating certain conditions
described herein, since the additional oxide may have or contribute to
an undesirable side effect, for example, such as skin irritation when
applied topically. For those embodiments, the compositions minimize
the amount of additional oxidizing agent, such as persulfate, or are
substantially or completely free of added persulfates or other
oxidizing agents.

Certain of the electron active metal oxides may be black in color,
such that care must be taken when formulating suitable topical
pharmaceutical compositions according to the invention to inhibit
blackening or superficial discoloration of the skin. Without being
bound by theory, it is believed that larger amounts of such
compositions promote increased superficial discoloration. Thus, in one
embodiment, the pharmaceutical compositions preferably have an
insufficient amount of metal oxide composition to cause visible skin
discoloration.

Additionally, it was found by rigorous testing that certain silver
tetroxide-containing compositions were comparatively non-toxic
compared to silver salts, such as conventional formulations of silver
nitrate, silver sulfadiazine, and benzoyl peroxide. Since these silver
tetroxide compositions were effective at certain ppm concentrations in
killing pathogens in nutrient broth and for water treatment,
commercial concentrates were formulated with 2% of the tetrasilver
tetroxide. For acceptance of the oxide in commerce, for which EPA
registration No. 3432-64 was obtained, it was necessary for the Ag.sub.
4 O.sub.4 to undergo a series of toxicity tests. A 3% concentrate was
used and evaluated by a certified laboratory employing good laboratory
practice (GLP) according to the Code of Federal Regulations for this
purpose. The results were as follows:

Acute Oral Toxicity LD.sub.50 Greater than 5,000 mg/Kg Acute Dermal
Toxicity LD.sub.50 Greater than 2,000 mg/Kg Primary Eye Irritation
Mildly irritating Primary Skin Irritation No irritation Skin
Sensitization Non-Sensitizing

Subsequent evaluations conducted according to the invention showed
that unless persons were prone to silver allergies, the pure
tetrasilver tetroxide compositions according to the invention could be
applied to the skin without any ill effects or evidence of irritation,
despite the fact that the compositions of the invention can be a
powerful oxidizing agent. This can perhaps be explained by the
stability manifested by the K.sub.A of the tetrasilver tetroxide
compositions, which is approximately 7.9.times.10.sup.-13.

Where the electron active compositions according to the invention are
applied to the skin, they may be combined with a carrier in an amount
from about 5 ppm to 500,000 ppm, more preferably from about 50 ppm to
250,000 ppm of the electron active metal oxide composition, based on
the weight of the composition. In various embodiments, the
compositions are provided in amounts from about 400 ppm to 100,000
ppm, from about 1,000 ppm to 70,000 ppm, from about 10,000 ppm to
50,000 ppm, or from about 20,000 ppm to 40,000 ppm, based on the
weight of the composition. In one preferred embodiment, the
compositions are formulated with about 25,000 ppm to 35,000 ppm of
metal oxide, based on the weight of the composition. It will be
readily understood by those of ordinary skill in the art that the ppm
concentration of electron active compound(s), such as metal oxide, in
the composition is based on the total weight of the composition.

When prevent, treating, or managing conditions, a preferred embodiment
employs amounts of about 0.1 to 10 percent by weight, about 0.25 to 5
percent by weight, or about 2 to 4 percent by weight of the compounds
or compositions of the invention. The compositions, when applied
topically, can be applied to the skin about1 to 3 times per day until
the condition is suitably cured or satisfactorily controlled. In one
embodiment, the composition may generally be topically applied at a
dosage level of from about 1 mg to 1000 mg per cm.sup.2 of skin
surface, preferably about 10 mg to 500 mg per cm.sup.2 of skin
surface. When applied topically, a preferred carrier includes
petroleum jelly, such as white petroleum jelly. For example, a
suitable white petroleum jelly is available from Penreco of Houston,
Tex.

Most of the metal oxide compounds for use according to the invention
are commercially available from various sources. Tetrasilver tetroxide
compositions for use according to the invention have been commercially
sold under the poorly named "Ag(II) OXIDE" tradename. They may be
obtained from Aldrich Chemical Co., Inc., having a place of business
in Milwaukee, Wis. The chemical synthesis of tetrasilver tetroxide
compounds can be performed according to the method described on page
148 in M. Antelman, "Anti-Pathogenic Multivalent Silver Molecular
Semiconductors," Precious Metals, vol. 16:141-149 (1992) by reacting
silver nitrate with potassium peroxydisulfate according to the
following equation in alkali solutions:

To the extent necessary to understand the present invention, the
disclosure of Antelman is hereby incorporated herein by express
reference thereto.

Tetracopper tetroxide, also referred to herein as Cu(I,III) oxide or
Cu.sub.4 O.sub.4, is a preferred electron active compound in
accordance with the invention. This compound may be prepared as
follows.

Suitable copper-based starting materials for this reaction include at
least one copper(I)-containing material. In one embodiment, a water
soluble copper(I) salt can be used. Typically, a water soluble
copper(I) salt can be prepared by dissolving an inorganic copper(I)
compound, for example, such as cuprous oxide, in an appropriate acid,
for example, an organic acid, such as acetic acid. Since soluble
copper(I) salts are not readily commercially available at the present
time, however, a non-solvated inorganic copper(I) compound, such as
cuprous oxide itself, can be used as the copper(I)-containing starting
material. In addition, other copper(I)-containing materials, either
inorganic, such as a copper(I) oxide, or organic, such as an
organometallic copper(I) compound, or both, may be used, where the
copper(I)-containing material(s) are sufficiently soluble in an
aqueous or organic solution to allow reaction with other materials to
form an electron active copper oxide compound.

The copper(I)-containing starting material is combined with an aqueous
caustic solution. This caustic solution preferably contains two
components: a strong caustic base and a peroxy acid salt. Examples of
suitable strong caustic bases include Group I and Group II hydroxides,
preferably sodium hydroxide or potassium hydroxide. Examples of
suitable peroxy acid salts include Group I salts of persulfates,
preferably potassium persulfate.

The copper-based starting material is typically the limiting reagent
in such a preparation. The ratio of each of the components in the
caustic solution to that of the copper-based starting material is
theoretically set by the stoichiometry of the particular reaction. In
one preferred embodiment, there is a relative molar excess, i.e., an
amount more than stoichiometrically necessary, of each of the
components in the caustic solution with respect to the copper-based
starting material. When a strong caustic base and a peroxy acid salt
are present in the caustic solution, the relative molar excesses of
the components may be at least about 50% and at least about 10%,
respectively, preferably at least about 100% and at least about 20%,
respectively, more preferably, at least about 250% and at least about
40%, respectively, most preferably at least about 500% and at least
about 75%, respectively.

Generally, the reactants may be added together in any manner that
comports with typical laboratory procedure. In one embodiment, the
copper(I)-containing starting material is placed in a reactor, to
which the strong caustic base and the peroxy acid salt are added, each
typically in their own solutions. The solution containing the
reactants is then typically heated to a temperature sufficient to
activate a reaction, preferably sufficient to activate a reaction with
no major undesirable side reactions or other undesirable effects, more
preferably above about 80.degree. C., most preferably about 90.degree.
C. to 95.degree. C. The solution is heated for a time sufficient to
facilitate the reaction, preferably to provide substantial completion
of the reaction, preferably for at least about 5 minutes, more
preferably for at least about 15 minutes, after which time the
solution is allowed to cool or is cooled, preferably to below about
45.degree. C., more preferably to about room temperature.

The color change of the solution, from its original color, red, to a
color indicating a reaction has occurred, in this case black, may
occur at the heated temperature or during or after cooling.

The purification and isolation of the desired product can be
accomplished by any suitable method available to those of ordinary
skill in the art. In the majority of situations, the desired reaction
product is primarily a solid, but may be dissolved or dispersed in at
least part of the solution. In one preferred embodiment, the solution
is carefully decanted off, and then the remaining product is washed
multiple times with distilled water, before being sufficiently dried.
In another preferred embodiment, the solution is vacuum filtered to
remove the filtrate, and the remaining product is sufficiently dried.

The yield of solid tetracopper tetroxide material, based on the
reactants, is typically at least about 10%, preferably at least about
45%, more preferably at least about 75%, most preferably at least
about 80%.

In addition, Fe(II,III) oxide and Mn(II,III) oxide are commercially
available from Aldrich Company of Milwaukee, Wis., and Co(II,III)
oxide and Pr(III,IV) oxide are commercially available from Noah
Technologies of San Antonio, Tex. Also, Bi(III,V) oxide synthetic
routes are detailed and reviewed in Gmelins Handbuch Der Anorganischen
Chemie, vol. 16:642 (1964), and the oxide is available commercially
from City Chemicals of New York, N.Y.

The magnitude of a prophylactic or therapeutic dose of electron active
composition(s), or a derivative thereof, in the acute or chronic
management of diseases and disorders described herein will vary with
the severity of the condition to be prevented, treated, or managed and
the route of administration. For example, oral, mucosal (including
rectal and vaginal), parenteral (including subcutaneous,
intramuscular, bolus injection, and intravenous, such as by infusion),
sublingual, transdermal, nasal, buccal, and like may be employed. In
one embodiment, a patient may gargle using the composition of the
present invention. Dosage forms include tablets, troches, lozenges,
dispersions, suspensions, suppositories, solutions, capsules, soft
elastic gelatin capsules, patches, and the like. The dose, and perhaps
the dose frequency, will also vary according to the age, body weight,
and response of the individual patient. Suitable dosing regimens can
be readily selected by those of ordinary skill in the art with due
consideration of such factors. In general, the total daily dosage for
the conditions described herein, is from about 0.1 mg to 1,000 mg of
the active ingredient, i.e., one of the metal oxides described herein,
or a derivative thereof. In another embodiment, the daily dosage can
be from about 1 mg to 500 mg, while in another embodiment, the daily
dosage can be from about 2 mg to 200 mg of the metal oxide
composition. A unit dosage can include, for example, 30 mg, 60 mg, 90
mg, 120 mg, or 300 mg of metal oxide composition. Preferably, the
active ingredient is administered in single or divided doses from one
to four times a day, such as by topical administration. In another
embodiment, the compositions are administered by an oral route of
administration. The oral dosage forms may be conveniently presented in
unit dosage forms and prepared by any methods available to those of
ordinary skill in the art of pharmacy.

In managing the patient, the therapy may be initiated at a lower dose,
e.g., from about 1 mg, and increased up to the recommended daily dose
or higher depending on the patient's global response. It is further
recommended that children, patients over 65 years, and those with
impaired renal or hepatic function, initially receive low doses when
administered systemically, and that they be titrated based on
individual response(s) and blood level(s). It may be necessary to use
dosages outside these ranges in some cases, as will be apparent to
those of ordinary skill in the art. Furthermore, it is noted that the
clinician or treating physician will know how and when to interrupt,
adjust, or terminate therapy in conjunction with individual patient
response.

Any suitable route of administration may be employed for providing the
patient with an effective dosage of electron active metal oxide, or a
derivative thereof. The most suitable route in any given case will
depend on the nature and severity of the condition being prevented,
treated, or managed.

In practical use, the metal oxide, or a derivative thereof, can be
combined as the active ingredient in intimate admixture with a
pharmaceutical carrier according to conventional pharmaceutical
compounding techniques. The carrier may take a wide variety of forms
and may include a number of components depending on the form of
preparation desired for administration. The compositions of the
present invention may include, but are not limited to, suspensions,
solutions and elixirs; aerosols; or carriers, including, but not
limited to, starches, sugars, microcrystalline cellulose, diluents,
granulating agents, lubricants, binders, disintegrating agents, and
the like.

Suitable forms in which the electron active compounds or compositions
of the present invention may be used include, but are not limited to,
powder, granule, flake, solution, suspension, emulsion, slurry,
aerosol spray, gel, paste, and combinations thereof. In one preferred
embodiment, the form is a powder or solution. When the electron active
compounds are in the form of a solution, the solution may be aqueous,
non-aqueous, or a combination thereof, preferably at least partially
aqueous, more preferably substantially aqueous. In a preferred
embodiment, the metal oxides are in an aqueous solution.

The compositions of the invention may be applied topically, e.g.,
either directly as a powder or in non-sprayable or sprayable form. Non-
sprayable forms can be semi-solid or solid forms including a carrier
indigenous to topical application and preferably having a dynamic
viscosity greater than that of water. Suitable formulations include,
but are not limited to, suspensions, emulsions, creams, ointments,
powders, liniments, salves and the like. If desired, these may be
sterilized or mixed with any available auxiliary agents, carriers, or
excipients, e.g., thixotropes, stabilizers, wetting agents, and the
like. One or more thixotropic agents can be included in types and
amounts sufficient to increase adhesion of topically applied
compositions of the invention to the skin, so as to inhibit or prevent
runoff or other loss of the composition from the treatment zone on the
skin. Preferred vehicles for non-sprayable topical preparations
include ointment bases, e.g., polyethylene glycol-1000 (PEG-1000);
conventional ophthalmic vehicles; creams; and gels, as well as
petroleum jelly and the like. In one more preferred embodiment, the
carrier includes a petroleum jelly. In another preferred embodiment,
the carrier is formulated as a cream, gel, or lotion. In another
preferred embodiment, the carrier is 3 weight percent active
ingredient, 36 weight percent heavy mineral oil, 47 weight percent
petroleum jelly, and 14 weight percent Tivawax P, which is available
from Tivian Laboratories, Inc., of Providence, R.I. In yet another
preferred embodiment, the composition may be a dry powder, such as
with 5 weight percent active ingredient and 95 weight percent bismuth
subgallate. These topical preparations may also contain emollients,
perfumes, and/or pigments to enhance their acceptability for various
usages.

The compositions may also be formulated for parenteral administration
by injection (subcutaneous, bolus injection, intramuscular, or
intravenous, such as by infusion), and may be dispensed in a unit
dosage form, such as a multidose container or an ampule. Compositions
of the electron active metal oxide, or a derivative thereof, for
parenteral administration may be in the form of suspensions,
solutions, emulsions, or the like, in aqueous or oily vehicles, and in
addition to the active ingredient may contain one or more formulary
agents, such as dispersing agents, suspending agents, stabilizing
agents, preservatives, and the like.

In the case where an intravenous injection or infusion composition is
employed, a suitable dosage range can be, e.g., from about 0.5 mg (0.1
ppm) to about 1,000 mg (200 ppm) total dose, preferably from about 5
mg (1 ppm) to 400 mg (80 ppm). In one preferred embodiment, the total
dose can be from about 50 mg (10 ppm) to 200 mg (40 ppm). It should be
understood that any suitable amount of the composition according to
the invention may be administered if effective to prevent, treat, or
manage one or more conditions described herein.

Pharmaceutical compositions of the present invention may be orally
administered in discrete pharmaceutical unit dosage forms, such as
capsules, cachets, soft elastic gelatin capsules, tablets, or aerosols
sprays, each containing a predetermined amount of the active
ingredient, as a powder or granules, or as a solution or a suspension
in an aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion,
or a water-in-oil liquid emulsion. Such compositions may be prepared
by any of the methods of pharmacy, but all methods include the step of
bringing into association the active ingredient with the
pharmaceutically acceptable carrier which constitutes one or more
necessary ingredients. In general, the compositions are prepared by
uniformly and intimately admixing the active ingredient with liquid
carriers or finely divided solid carriers or both, and then, if
necessary, shaping the product into the desired presentation. Suitable
types of oral administration include oral solid preparations, such as
capsules or tablets, or oral liquid preparations. If desired, tablets
may be coated by standard aqueous or nonaqueous techniques.

For example, a tablet may be prepared by compression or molding,
optionally, with one or more accessory ingredients. Compressed tablets
may be prepared by compressing in a suitable machine the active
ingredient in a free-flowing form such as powder or granules,
optionally mixed with a binder, lubricant, inert diluent, granulating
agent, surface active agent, dispersing agent, or the like. Molded
tablets may be made by molding, in a suitable machine, a mixture of
the powdered compound moistened with an inert liquid diluent. In one
embodiment, each tablet, capsule, cachet, or gel cap contains from
about 0.5 mg to about 500 mg of the active ingredient, while in
another embodiment, each tablet contains from about1 mg to about 250
mg of the active ingredient. The amount of active ingredient found in
the composition, however, may vary depending on the amount of active
ingredient to be administered to the patient.

Another suitable route of administration is transdermal delivery, for
example, via an abdominal skin patch.

The metal oxide(s), or a derivative thereof, may be formulated as a
pharmaceutical composition in a soft elastic gelatin capsule unit
dosage form by using conventional methods well known in the art, such
as in Ebert, Pharm. Tech, 1(5):44-50 (1977). Soft elastic gelatin
capsules have a soft, globular gelatin shell somewhat thicker than
that of hard gelatin capsules, wherein a gelatin is plasticized by the
addition of plasticizing agent, e.g., glycerin, sorbitol, or a similar
polyol. The hardness of the capsule shell may be changed by varying
the type of gelatin used and the amounts of plasticizer and water. The
soft gelatin shells may contain an additional preservative, such as
methyl- and propylparabens and sorbic acid, to prevent the growth of
fungi, although this is not necessary since the compounds and
compositions of the invention provide anti-fungal efficacy. Thus, in
one embodiment, the invention includes a compositions formulated as a
gelatin shell with an electron active metal oxide compound of the
present invention, completely free of added preservatives. The active
ingredient may be dissolved or suspended in a liquid vehicle or
carrier, such as vegetable or mineral oils, glycols such as
polyethylene glycol and propylene glycol, triglycerides, surfactants
such as polysorbates, or a combination thereof.

In addition to the common dosage forms set out above, the compounds of
the present invention may also be administered by controlled release
means, delivery devices, or both, as are well known to those of
ordinary skill in the art, such as those described in U.S. Pat. Nos.
3,845,770; 3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533;
5,059,595; 5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; and
5,733,566, the disclosures of which are hereby incorporated herein by
express reference thereto. These pharmaceutical compositions can be
used to provide slow or controlled-release of the active ingredient
therein using, for example, hydropropylmethyl cellulose in varying
proportions to provide the desired release profile, other polymer
matrices, gels, permeable membranes, osmotic systems, multilayer
coatings, microparticles, liposomes, microspheres, or the like, or a
combination thereof. Suitable controlled-release formulations
available to those of ordinary skill in the art, including those
described herein, may be readily selected for use with the tetrasilver
tetroxide compositions of the invention. Thus, single unit dosage
forms suitable for topical or oral administration, such as gels,
lotions, cremes, tablets, capsules, gelcaps, caplets, and the like,
that are adapted for controlled-release are encompassed by the present
invention.

All controlled-release pharmaceutical products have a common goal of
improving drug therapy over that achieved by their non-controlled
counterparts. Ideally, the use of an optimally designed controlled-
release preparation in medical treatment is characterized by a minimum
of the active ingredient being employed to cure or control the
condition in a minimum amount of time. Advantages of controlled-
release formulations may include: 1) extended activity of the active
ingredient; 2) reduced dosage frequency; and 3) increased patient
compliance.

Most controlled-release formulations are designed to initially release
an amount of active ingredient that promptly produces the desired
therapeutic effect, and gradual and continual release of other amounts
of active ingredient to maintain this level of therapeutic effect over
an extended period of time. In order to maintain this constant level
of active ingredient in the body, the active ingredient should be
released from the dosage form at a rate that will replace the amount
of active ingredient being metabolized and excreted from the body.

The controlled-release of the active ingredient may be stimulated by
various inducers, for example pH, temperature, enzymes, water, or
other physiological conditions or compounds. The term "controlled-
release component" in the context of the present invention is defined
herein as a compound or compounds, including polymers, polymer
matrices, gels, permeable membranes, liposomes, microspheres, or the
like, or a combination thereof, that facilitates the controlled-
release of the active ingredient (e.g., tetrasilver tetroxide) in the
pharmaceutical composition.

The pharmaceutical compositions for use in the present invention
include electron active metal oxides, or a derivative thereof, as the
active ingredient, and may also contain a pharmaceutically acceptable
carrier, and optionally, other therapeutic ingredients. Suitable
derivatives include any available "pharmaceutically acceptable salts,"
which refer to a salt prepared from pharmaceutically acceptable non-
toxic acids including inorganic acids, organic acids, solvates,
hydrates, or clathrates thereof. Examples of such inorganic acids are
nitric, sulfuric, lactic, glycolic, salicylic, and phosphoric.
Appropriate organic acids may be selected, for example, from
aliphatic, aromatic, carboxylic and sulfonic classes of organic acids,
examples of which are formic, acetic, propionic, succinic,
camphorsulfonic, citric, fumaric, gluconic, isethionic, lactic, malic,
mucic, tartaric, para-toluenesulfonic, glycolic, glucuronic, maleic,
furoic, glutamic, benzoic, anthranilic, salicylic, phenylacetic,
mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic,
pantothenic, benzenesulfonic (besylate), stearic, sulfanilic, alginic,
galacturonic, and the like. Particularly preferred acids are lactic,
glycolic, and salicylic acids. The pharmaceutically acceptable salts
preferably do not include halide-containing salts when tetrasilver
tetroxide is present, as these salts are believed to facilitate
breakdown of the oxide lattice present in the silver oxide
compositions of the invention.

EXAMPLES

These and other aspects of the present invention may be more fully
understood with reference to the following non-limiting examples,
which are merely illustrative of the preferred embodiments of the
present invention, and are not to be construed as limiting the
invention, the scope of which is defined by the appended claims.

EXAMPLES 1-16

Antipathogenic Efficacy of Compositions

EXAMPLES 1-2

In Vitro Treatment of Salmonella with Compositions of Invention

A culture of Salmonella of cell density 500,000 CFU/mL was contacted
for 10 minutes with approximately 4 ppm hexapraseodymium undecoxide
(Pr.sub.6 O.sub.11), which is believed to contain two distinct
oxidation states of praseodymium, Pr(III) and Pr(IV), in its crystal
lattice, at a pH of about 9, followed with a culture adjusted to a pH
of about 10. The percentages of bacterial colonies killed by this
treatment were 96.4% and 93.8%, respectively. The experiment was
repeated with the same cell density of Salmonella using about 5 ppm of
tricobalt tetroxide (Co.sub.3 O.sub.4), which is believed to contain
two distinct oxidation states of cobalt, Co(II) and Co(III), in its
crystal lattice, at a pH of about 10. The percentage of bacterial
colonies killed by this treatment was 92.8% after 10 minutes of
contact with the oxide-containing composition.

EXAMPLES 3-4

Antipathozenic Effect of Compositions in Water Purification

The praseodymium oxide crystals of Example 1 were tested against the
standard AOAC coliform culture used in water purification studies and
having a 375,000 CFU/mL density. The results of this study are
tabulated below:

ppm Pr.sub.6 O.sub.11 Contact time (mins.) pH Bacteria mortality (%) 4
5 7 68 4 10 7 71 10 5 7 65 10 10 7 76 5 5 9 84 10 10 9 88

The experiment of Example 3 was repeated with about 4 ppm of tricobalt
tetroxide (Co.sub.3 O.sub.4) according to Example 2, at a contact time
of about 5 minutes at a pH of about 7. The percentage of bacterial
colonies killed by this treatment was 75%.

EXAMPLE 5

In vitro Treatment of Stayhylococcus aureus with Compositions

1 gram of Pr (III,IV) oxide (Pr.sub.6 O.sub.11) was dissolved in 20 mL
of 85% phosphoric acid, which underwent substantially no redox
reaction with the praseodymium oxide, such that an active solution was
formed. The solution was subsequently diluted to yield a 100 ppm
solution, based on the oxide component. The Pr (III,IV) oxide
solution, when put in contact with Staphylococcus aureus at 220,000
CFU/mL cell density, served to kill substantially all the bacteria
(100% mortality) after 10 minutes of contact with the oxide-containing
composition.

EXAMPLES 6-9

In vitro Treatment of E. coli with Compositions of the Invention

A culture of E. coli bacteria having a cell density of 420,000 CFU/mL
was contacted for about 10 minutes with about 6 ppm of Co (II,III)
oxide, Co.sub.3 0.sub.4, at a pH of about 7, also in the presence of
10 ppm potassium monopersulfate, which is commercially available under
the trademark OXONE from DuPont De Nemours, Inc., of Wilmington, Del.
The percentage of bacteria killed by this contact was 47.6%. When
repeating the previous experiment using a culture having a cell
density of 380,000 CFU/mL and with about 5 ppm of Pr (III,IV) oxide in
the presence of about 50 ppm OXONE.TM., the percentage of bacteria
killed was 39.5%.

A culture of E. coli bacteria having a cell density of 160,000 CFU/mL
was contacted for about 10 minutes with about 100 ppm of Cu (1,111)
oxide, Cu.sub.4 O.sub.4. The percentage of bacteria killed by this
contact was 63.8%. When repeating the previous experiment using only
about half the Cu (I,III) oxide concentration, i.e., about 50 ppm, in
the presence of about 200 ppm OXONE.TM., the percentage of bacteria
killed was 97.8%.

EXAMPLES 10-13

In vitro Treatment of E. coli with Compositions of the Invention

Cultures of E. coli bacteria, each having a cell density around
100,000 CFU/mL, were each contacted for about 10 minutes with various
electron active molecular metal oxide crystals according to the
invention, resulting in the following percentages of bacteria killed:

Composition of the Invention % Bacteria Killed Bi (III,V) oxide,
Bi.sub.2 O.sub.4 38% Fe (II,III) oxide, Fe.sub.3 O.sub.4 32% Mn
(II,III) oxide, Mn.sub.3 O.sub.4 28%

These experiments were repeated using reduced triiron tetroxide,
Fe.sub.3 O.sub.4, concentrations and E. coli cultures, each having a
reduced cell density of 75,000 CFU/mL, with variable OXONE.TM.
concentrations. When Fe (II, III) oxide was used in about 50 ppm
concentration in the presence of about 200 ppm OXONE.TM., the
percentage of bacteria killed was about 73.3%. When Fe (II,III) oxide
was used in about 20 ppm concentration, in the presence of about 100
ppm OXONE.TM., the percentage of bacteria killed was about 49.3%.

EXAMPLES 14-16

In vitro Treatment of Staphylococcus aureus Using Compositions

Compositions containing about 100 ppm of Bi (III,V) oxide, Bi.sub.2
O.sub.4, Fe (II,III) oxide, Fe.sub.3 O.sub.4, or Mn (II,III) oxide,
Mn.sub.3 O.sub.4, were tested for antimicrobial efficacy by contacting
cultures of Staphylococcus aureus bacteria having cell densities of
75,000 CFU/mL for about 10 minutes. The iron and manganese oxide
compositions were observed to kill substantially no bacteria, whereas
the composition containing dibismuth tetroxide was observed to kill
about 37.3% of the bacteria.

EXAMPLE 17

Preparation of Tetracopper Tetroxide

2.4 grams each of sodium hydroxide and potassium persulfate were
dissolved, each in 25 mL of distilled water, each in its own 50 mL
beaker. These solutions were mixed together in another beaker, to
which 700 mg of red cuprous oxide was added. This beaker was heated to
approximately 90.degree. C. and was maintained from about 90.degree.
C. and 95.degree. C. for about 15 minutes before being allowed to cool
to room temperature. The heating of the solution caused a color change
from red to black, indicating a reaction of the oxide.

The solid product was purified and isolated by one of two methods: a)
decanting off the solution, washing the remaining product at least
seven times with distilled water, and drying the product; or b) vacuum
filtering the solution and drying the product. The experimental yield
was similar using either isolation method.

The average theoretical yield of Cu(I,III) oxide, or Cu.sub.4 0.sub.4,
was 83%, based on the following equation:

Based on all of the test data described above, the healing mechanism
associated with the use of the metal oxides of the invention to treat
and manage at least some skin diseases, without being bound by theory,
appears to involve mechanisms other than merely inhibiting or killing
pathogens and curing infections that tend to aggravate disease and
retard the natural healing process. The data indicate that healing is
brought about even in cases where no abnormal bacteria counts or
infection is evident. This suggests that the electron active
compound(s) may also act against auto-antibodies that trigger
autoimmune reactions associated with diseased tissue, as well as
against other non-pathogenic conditions or diseases, such as
circulatory or neurological conditions or diseases.

Although preferred embodiments of the invention have been described in
the foregoing Detailed Description, it will be understood that the
invention is not limited to the embodiments disclosed, but is capable
of numerous rearrangements and modifications of parts and elements
without departing from the spirit of the invention. It will be
understood that the chemical and pharmaceutical details of every
design may be slightly different or modified by one of ordinary skill
in the art without departing from the compositions and methods taught
by the present invention.



United States Patent 6,485,755
Antelman November 26, 2002
Methods of using electron active compounds for managing cancer

Abstract

The present invention provides methods for preventing, treating, and/
or managing one or more cancerous conditions in a patient, such as a
human. A multivalent metal oxide, such as Ag(I,III), Cu(I,III),
Pr(III,IV), and Bi(III,V) oxides or a pharmaceutically acceptable
derivative thereof, may be administered to the patient in an amount
and for a period of time which is therapeutically effective to
prevent, treat, and/or manage such condition(s). These cancerous
conditions include systemic and external cancers, and may also include
conditions and symptoms associated with cancer. The present invention
also provides a pharmaceutical composition suitable for treating such
cancerous conditions. The compositions of the invention may be adapted
for at least one of subcutaneous injection, intramuscular injection,
intravenous injection, infusion, transdermal, or topical application.
Inventors: Antelman; Marvin S. (Rehovot, IL)
Assignee: Marantech Holding (Providence, RI)
Appl. No.: 09/692,488
Filed: October 20, 2000
Related U.S. Patent Documents

Application Number Filing Date Patent Number Issue Date
552172 Apr., 2000 6258385

Current U.S. Class: 424/618 ; 424/600; 424/617; 424/630; 424/635;
424/639; 424/646; 424/653; 514/788.1
Current International Class: A61K 47/02 (20060101); A61K 8/02
(20060101); A61K 8/19 (20060101); A61Q 1/12 (20060101); A61Q 19/00
(20060101); A61K 33/24 (20060101); A61K 33/26 (20060101); A61K 33/32
(20060101); A61K 33/34 (20060101); A61K 33/38 (20060101); A61K 9/06
(20060101); A61K 033/38 (); A61K 033/00 (); A61K 047/00 (); A61K
047/30 ()
Field of Search: 424/600,617,618,630,639,646,647,648,653,635
514/788.1
References Cited [Referenced By]
U.S. Patent Documents

3923982 December 1975 Lamand et al.
4447254 May 1984 Hughes et al.
4574782 March 1986 Borrelli et al.
4735796 April 1988 Gordon
4828832 May 1989 De Cuellar et al.
4952411 August 1990 Fox, Jr. et al.
5017295 May 1991 Antelman
5073382 December 1991 Antelman
5078902 January 1992 Antelman
5089275 February 1992 Antelman
5098582 March 1992 Antelman
5211855 May 1993 Antelman
5223149 June 1993 Antelman
5320906 June 1994 Eley et al.
5334588 August 1994 Fox, Jr. et al.
5336416 August 1994 Antelman
5336499 August 1994 Antelman
5571520 November 1996 Antelman
5612019 March 1997 Gordon et al.
5676977 October 1997 Antelman
5772896 June 1998 Denkewicz, Jr. et al.
5928958 July 1999 Pilgrimm
Foreign Patent Documents

2000060976 Feb., 2000 JP

Other References

STN/CAS online, file CIN, Acc. No. 13(8):6866B, China Dly. (North Am.
Ed.), Jan. 30, 1984, p. 5), Abstract.* .
Antelman, Marvin S.; "Silver (II,III) Disinfectants"; Soap/Cosmetics/
Chemical Specialties, Mar. 1994, pp. 52-59. .
Antelman, Marvin S.; Abstracts of the American Chemical Society;
1992(203). .
Antelman, Marvin S.; "Anti-Pathogenic Multivalent Silver Molecular
Semiconductors"; Precious Metals; 1992(16); pp. 141-149. .
Antelman, Marvin S.; "Multivalent Silver Bactericides"; Precious
Metals; 1992(16); pp. 151-163. .
Fung, Man C. and Bowen, Debra L.; "Silver Products for Medical
Indications: Risk-Benefit Assessment", Clinical Toxicology, 1996, pp.
119-126. .
Dorland et al., Dorland's Illustrated Medical Dictionary,
Philadelphia: W.B. Saunders Company, 1994, 28.sup.th Edition, p. 351,
759, and 760. .
Gennaro, A., Remington's Pharmaceutical Sciences, Easton, PA: Mack
Publishing Company, 1985, 17.sup.th Edition, p. 1573-1575, 1585-1594,
and 1601..

Primary Examiner: Dees; Jose' G.
Assistant Examiner: Choi; Frank
Attorney, Agent or Firm: Pennie & Edmonds LLP
Parent Case Text


CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation-in-part of application Ser. No.
09/552,172, filed Apr. 18, 2000, now U.S. Pat. No. 6,258,385, and
claims the benefit of Provisional Application No. 60/174,793, filed
Jan. 6, 2000, No. 60/184,053, filed Feb. 22, 2000, and No. 60/214,503,
filed Jun. 28, 2000.
Claims


What is claimed is:

1. A method for preventing, treating, or managing one or more
cancerous conditions or dysplastic proliferations in an animal, which
method comprises: administering at least one metal oxide compound
selected from the group consisting of Bi(III,V) oxide, Co(II,III)
oxide, Cu(I,III) oxide, Mn(II,III) oxide, Pr(III,IV) oxide, and
Ag(I,III) oxide to the animal in an amount and for a period of time
which is therapeutically effective to treat such condition(s).

2. The method of claim 1, wherein the metal oxide compound is
administered via intravenous injection or infusion and the animal is a
human.

3. The method of claim 2, wherein the administering is subcutaneous,
intramuscular, or by infusion into a blood stream of the animal.

4. The method of claim 3, wherein the metal oxide compound is
administered via infusion over a period of from about 30 minutes to
about 300 minutes to inhibit adverse side effects.

5. The method of claim 4, wherein the at least one other
chemotherapeutic agent is administered concurrently with the metal
oxide compound.

6. The method of claim 2, wherein the cancer includes skin cancer that
has metastasized.

7. The method of claim 1, wherein the metal oxide compound is
administered via intravenous injection or infusion and the animal is a
human.

8. The method of claim 7, wherein the metal oxide compound is
administered in an amount sufficient to provide about 1 to about 75
ppm of the metal oxide compound in the bloodstream.

9. The method of claim 1, wherein the metal oxide compound is
administered in conjunction with at least one other chemotherapeutic
agent.

10. The method of claim 1, wherein the cancerous condition or
dysplastic proliferation includes at least one of colon cancer, lung
cancer, throat cancer, breast cancer, kidney cancer, pancreatic
cancer, bladder cancer, prostate cancer, uterine cancer, brain cancer,
liver cancer, skin cancer, testicular cancer, stomach cancer, adrenal
gland cancer, cancer of the ovaries, thyroid cancer, bronchial cancer,
trachea cancer, eye cancer, bone cancer, cervical cancer, oral cavity
cancer, soft tissue cancer, pituitary gland cancer, myeloma, rectal
cancer, esophageal cancer, leukemia, lymphoma, cancerous fibroid
tumors, non-cancerous fibroid tumors, or liver cancer.

11. The method of claim 10, wherein the controlled release vehicle is
implanted in the body at a location suitable for providing a
therapeutically effective amount of metal oxide compound to the
patient without affecting proper functioning of the animal's liver.

12. The method of claim 1, wherein the metal oxide compound is
administered by a controlled release vehicle.

13. The method of claim 1, wherein the metal oxide compound is
substantially free of added persulfate.

14. A method for preventing, treating, or managing one or more
cancerous conditions or dysplastic proliferations associated with a
patient's skin, which method comprises administering the at least one
metal oxide compound selected from the group consisting of Bi(III,V)
oxide, Co(II,III) oxide, Cu(I,III) oxide, Mn(II,III) oxide, Pr(III,IV)
oxide, and Ag(I,III) oxide to the skin in an amount and for a period
of time which is therapeutically effective to treat such cancerous
condition(s).

15. The method of claim 14, wherein the at least one metal oxide
compound is substantially free of added persulfate.

16. The method of claim 14, wherein the cancerous condition or
dysplastic proliferation comprises at least one of dysplastic nevi,
neurofibromatosis, basal cell carcinoma, squamous carcinoma, or
melanoma.

17. The method of claim 14, wherein the cancerous condition or
dysplastic proliferation comprises symptoms of cancer or conditions
associated with a predisposition to cancer.

18. The method of claim 14, further comprising a carrier medium in
which the at least one metal oxide compound is dispersed, wherein the
therapeutically effective amount is from about 50 ppm to 500,000 ppm,
based on the weight of the carrier medium.

19. The method of claim 18, wherein the carrier medium comprises
petroleum jelly or mineral oil.

20. The method of claim 14, wherein the at least one metal oxide
compound is administered in the form of a powder.

21. The method of claim 14, wherein the therapeutically effective
amount is from about 400 ppm to 100,000 ppm.

22. The method of claim 14, wherein the administering is topical or
transdermal.

23. The method of claim 22, wherein the composition is topically
administered directly to the skin.

24. The method of claim 23, wherein the at least one metal oxide
compound further comprises a thixotropic agent sufficient to increase
adherence of the composition to the skin without excessive runoff.

25. The method of claim 14, wherein the administering comprises
application of the at least one metal oxide to the skin at a dosage
level of about 10 mg to 500 mg per cm.sup.2 of skin surface.

26. A method for preventing, treating, or managing one or more
cancerous conditions associated with a cervix of a female animal,
which method comprises administering at least one metal oxide compound
selected from the group consisting of Bi(III,V) oxide, Co(II,III)
oxide, Cu(I,III) oxide, Mn(III,II) oxide, Pr(III,IV) oxide, and
Ag(I,III) oxide to the cervix in an amount and for a period of time
which is therapeutically effective to treat such condition(s).

27. The method of claim 26, wherein the at least one metal oxide
compound is substantially free of added persulfate.

28. The method of claim 27, wherein the at least one metal oxide
compound is applied directly to the cervix.

29. The method of claim 28, further comprising a carrier medium in
which the at least one metal oxide compound is dispersed, wherein the
therapeutically effective amount is from about 50 ppm to 500,000 ppm,
based on the weight of the carrier medium.

30. The method of claim 29, wherein the carrier medium comprises
petroleum jelly.

31. The method of claim 26, wherein the at least one metal oxide
compound is applied in an amount sufficient to obtain a desired effect
and to substantially inhibit
Description


FIELD OF THE INVENTION

The invention relates to pharmaceutical compositions including at
least one metal oxide, such as an electron active metal oxide, and
methods of using such compositions, for the prevention, treatment, and
management of cancer and conditions or diseases related to the
presence of cancer or a predisposition to cancer.

BACKGROUND OF THE INVENTION

Cancers are a leading cause of death in animals and humans. The exact
cause of cancer is not known, but links between certain factors, such
as smoking or exposure to carcinogens including tobacco smoke and
chromium (VI), exposure to radiation, such as from x-rays,
radioisotopes, and ultra-violet light, viruses, such as papaloma,
Espstein Barr, and Raus sarcoma virus and the incidence of certain
types of cancers and tumors has been shown by a number of researchers.
Genetic factors and genome defects such as those found a chromosome 11
have also been linked to cancer. Traditional methods of cancer therapy
include treatment with chemotherapeutic agents that inhibit cell
division or radiation therapy that disrupts DNA in dividing cells.
These treatments, however, may also adversely affect normal cells that
happen to be dividing or synthesizing DNA at the time of treatment.
Dosage levels low enough to insure survival of a cancer patient often
are not sufficiently cytotoxic to tumor cells to retard continuing
cell division after treatment. Additionally, the mechanism for the
action of these chemotherapeutic agents is frequently unknown, which
complicates the safe and effective use of these agents. Several
different cancers and conditions associated with cancer are discussed
below as examples illustrating the importance of combating cancer and
associated conditions.

Breast carcinoma is the most common malignancy among women and shares
with lung carcinoma the highest fatality rate of all cancers affecting
females. For example, approximately one of every 11 women in the
U.S.A. will develop breast cancer. For white women, the probability is
about 1 in 10; for African American women, the rate is close to 1 in
14. The annual mortality rate from 1930 to the present has remained
fairly constant at about 27 deaths per 1000,000 females, and is
slightly higher for whites than African Americans.

In women, breast carcinoma is rare before age 30 but the incidence
rises rapidly after menopause. Post-menopausal breast masses are
typically considered cancerous until biopsy proves otherwise.
Cystosarcoma phyllodes, which are a non-cancerous tumor, are the most
common tumor of the breast; other malignancies are significantly more
rare. Breast cancer in men is rare and tends not to be recognized
until late with poor therapeutic results.

Most breast cancers, including those frequently designated as
scirrhus, infiltrative, papillary, ductile, medullary, and lobular,
appear as a slowly growing, painless mass, though a vague discomfort
may be present. Physical signs typically include a retracted nipple,
bleeding from the nipple, a distorted areola or breast contour, skin
dimpling over the lesion, attachment of the mass to surrounding
tissue, including the underlying fascia and overlying skin, edema of
the skin of the breast with an orange peel appearance, and axillary or
supraclavicular lymph nodes. In advanced cases, skin nodules with
ultimate breakdown and ulcer formation may be seen.

The presence of metastases should always be suspected as the disease
metastasizes by direct extension and via the lymph system and the
bloodstream. Among the most common sites are the lungs and pleura, the
skeleton (especially skull, spine, and pelvis), and the liver.
Although the exact causes of breast cancer are not known, a doctor
from France discovered a virus called mice breast tumor virus (vtmr)
in 1985 that was later described as an oncomavirus with particules
type B. This virus caused breast cancer in mice breast. It can be
transmitted by breast milk or it can be incorporated in the human
genome. Current treatments for breast cancer in general include
surgery, radiotherapy, chemotherapy and hormonal therapy.

Cervical cancer includes those cancer moieties which are indigenous to
the cervix. These cancer moieties are referred to generally as
cervical carcinomas of which 85-90% are squamous cell carcinomas, and
the balance are largely adenocarcinomas. The severity of cervical
cancers are gauged by the clinical tests called PAP smears which
indicate whether the carcinoma cells are confined to the cervix or
have penetrated beyond it but not to the pelvic wall, or to the pelvic
wall itself and even beyond the pelvis. Cervical cancers kill about
33% of their victims annually in the United States.

Carcinoma of the uterine cervix, the second most common malignancy of
the female reproductive tract, most commonly affects women aged 40 to
56 years old. The incidence is higher among women from lower
socioeconomic groups and among those with a history of early and
frequent coitus and multiple sexual partners. Recently, venereal
transmission of human papilloma virus (hpv) and herpes virus type 2
(nsv-2) have been implicated as important in the etiology of cervical
neoplasia.

The earliest histologic change in what is considered a continuum from
normal to invasive cancer is minimal cervical dysplasia, in which
abnormal cell proliferation occurs in the lower third of the
epithelium. Most of the minimal dysplasias are self-limiting and
regress to normal tissue. Most severe dysplasias in the upper two-
thirds of the epithelium showing abnormal proliferation, however,
progress to carcinoma in situ, in which a full thickness of the
epithelium contains abnormal calls. When cancer cells penetrate the
basement membrane and invade the stroma (invasive carcinoma) they can
spread by direct extension to adjacent pelvic organs or by lymphatic
permeation and dissemination.

Of cervical carcinomas, 85 to 90% are squamous cell carcinoma. These
vary from well-differentiated cells with keratinization to the highly
anaplastic spindle cells of cervical tumors. Adenocarcinomas, observed
in only 10 to 15% of cases, are more rare.

Early cervical neoplasia can be detected pre-clinically by cytologic
examination of cervical smears obtained during routine annual pelvic
examinations. At this stage, the disease is asymptomatic. The cervical
smears (pap test) can detect 90% of early cervical neoplasias. Thus,
the use of cervical smears has reduced the death rate from cervical
cancer by more than 50% through recognition and treatment of pre-
invasive neoplasia. Treatment of cervical cancer typically involves
conization, radiotherapy, surgical therapy, and chemotherapy.

For diagnostic and prognostic purposes, the results of cervical smear
tests may be grouped into four categories: class I characterized by
the absence of observed abnormal cells; class II characterized by the
presence of atypical cells and usually associated with inflammation;
class III characterized by the presence of cells representative of or
suspicious of carcinoma; and classes IV and V each characterized by
the presence of carcinoma cells.

Additionally, the clinical stage or progression of the cervical
carcinoma may be further characterized as follows. Stage 0 is
characterized by carcinoma in situ with intra epithelial carcinoma.
Stage I includes carcinomas strictly confined to the cervix. Stage IA
is characterized by micro invasive carcinoma and stage IB is
characterized by occult cancer.

In stage II, the carcinoma extends beyond the cervix but not onto the
pelvic wall. Stage IIA exhibits no obvious parametrial involvement
while stage IIB exhibits obvious parametrial involvement. In stage
III, the carcinoma extends onto the pelvic wall. Stage IIIA is
characterized by the lack of extension onto the pelvic wall and stage
IIIB is characterized by extension onto the pelvic wall.

In stage IV, the carcinoma has extended beyond the true pelvis or has
clinically involved the mucosa of the bladder. Stage IVA is
characterized by the spread of the growth to adjacent organs. Stage
IVB is characterized by the spread to distant organs.

Skin cancer is a disease in which cancer (malignant) cells are found
in the layers of the skin. The skin has two main layers and several
kinds of cells: a top layer called the epidermis, which contains three
kinds of cells: flat, scaly cells on the surface called squamous
cells; round cells called basal cells; and cells called melanocytes,
which give the skin its color. The dermis is the inner, second layer
of the skin.

The skin is the most environmentally-stressed organ in mammals,
particularly in humans. The skin is subjected to toxic chemicals and
hostile environments, as well as being the only organ directly exposed
to ultraviolet ("UV") light in the presence of oxygen. Lengthy
exposure of the skin to UV light typically damages the skin,
resulting, in sunburn, photo-aging, carcinogenesis, and other related
skin disorders. "Skin cancer" is generally used to describe the three
major forms of skin cancer; basal cell and squamous carcinoma together
with melanoma. These carcinomas account for about 97% of skin cancers.
Melanoma, however, accounts for over 87% of deaths due to said
cancers.

Melanoma is a disease of the skin in which cancer (malignant) cells
are associated with the cells that color the skin (melanocytes).
Melanoma usually occurs in adults, but it may occasionally be found in
children and adolescents. Melanoma is sometimes called cutaneous
melanoma or malignant melanoma. Melanoma can spread (metastasize)
quickly to other parts of the body through the lymph system or through
the blood.

About 80% of non-melanoma skin cancer will be basal cell carcinoma. It
can occur at any location on the body surface, but occurs more
commonly on sun-exposed surfaces, such as the face. The earliest sign
may be a red flat area, a small nodule, a small spot that bleeds on
rubbing, a small ulcer, or a scaly patch. About 20% of non-melanoma
skin cancer will be squamous cell carcinoma. The difference between
basal cell carcinoma and squamous cell carcinoma is often discernable
only at the microscopic level, as the two may look identical. Squamous
cell carcinoma, however, tends to grow more rapidly, and form an ulcer
sooner. Squamous cell carcinoma may afflict any skin surface, but is
common on the lips and ears.

Neurofibromatosis is a hereditary autosomal dominant disorder that is
accompanied by a predisposition to cancer. Neurofibromatosis produces
pigmented spots and tumors of the skin and of peripheral, optic and
acoustic nerves. Subcutaneous and bony deformities may also be
observed. One third of the patients with neurofibromatosis are
asymptomatic and the condition is discovered during routine
examination. In one-third of patients, cosmetic problems are the
initial complaints. Characteristic skin lesions, apparent at birth or
in infancy in 90% of the patients, include medium brown patches
distributed most commonly over the trunk, pelvis, and flexor creases
of elbows and knees. For diagnostic purposes, the presence of six or
more of these freckle-like lesions with one larger than 1.5 cm is
characteristic of neurofibromatosis. Multiple cutaneous tumors, flesh
colored and of variable size and shape typically appear in late
childhood.

The above-mentioned discussion merely illustrates the breadth and
importance of cancer as an affliction of animals and humans in
particular. Those of ordinary skill in the art will understand that
various other types of cancers exist that also require suitable
prevention, treatment, and/or management. In view of this discussion,
there is a need for pharmaceutical compositions that can be
administered at dosage levels low enough to insure survival of a
cancer patient but which are sufficiently cytotoxic to cancer cells or
cells associated with cancer to retard or eliminate continuing cell
division after treatment, i.e., management or treatment of the cancer.

Metal oxides, such as electron active metal oxides comprising
multivalent silver cations, have been disclosed for various uses, as
they are reported to be non-toxic to animals and humans. M. Antelman,
"Anti-Pathogenic Multivalent Silver Molecular Semiconductors,"
Precious Metals, vol. 16:141-149 (1992); M. Antelman, "Multivalent
Silver Bactericides," Precious Metals, vol. 16:151-163 (1992). For
example, tetrasilver tetroxide activated with an oxidizing agent is
disclosed for use in bactericidal, fungicidal, and algicidal use, such
as in municipal and industrial water treatment applications and for
the treatment of AIDS.

A variety of sources also report the use of certain divalent silver
compounds for water treatment, as well as the use of such compounds,
typically in combination with certain oxidizing agents, metals, or
other compounds, as disinfectants, bactericides, algicides, and
fungicides. One source also reports a single in vitro study of the use
of such compounds for the treatment of AIDS. These sources include M.
Antelman, "Silver (II, III) Disinfectants," Soap/Cosmetics/Chemical
Specialties, pp. 52-59 (March, 1994), and U.S. Pat. Nos. 5,017,295;
5,073,382; 5,078,902; 5,089,275; 5,098,582; 5,211,855; 5,223,149;
5,336,416; and 5,772,896.

U.S. Pat. No. 5,336,499 discloses tetrasilver tetroxide and persulfate
compositions having certain in vitro anti-pathogenic properties, i.e.,
bactericidal, fungicidal, viricidal, and algicidal, in certain
concentrations as low as 0.3 ppm, particularly in nutrient broth
cultures. The persulfate is disclosed to be an oxidizing agent that
activates the tetroxide crystals. Also disclosed are an in vitro study
regarding the inhibition of yeast growth in nutrient broth and the
formulation of a gynecological cream and douche based on these
results, and a report of an in vitro AIDS test with the compositions
indicating total suppression of the virus at 18 ppm.

In vitro assays, such as those disclosed in Ahmed, S. A., Gogal Jr.,
R. M. and Walsh, J. E., a New Rapid and Simple Non-radioactive Assay
to Monitor and Determine the Proliferation of Lymphocytes: an
Alternative to [.sup.3 H]-thymidine Incorporation Assay, Journal of
Immunological Methods 1994; 170: 211-224; Boyd, M. R., Status of the
NCI Preclinical Antitumor Drug Discovery Screen, J. B. Lippincott
Company, Philadelphia, Principles & Practices of Oncology Updates
1989; 3 # 10: 1-12, and Boyd, M. R. et al. Data Display and Analysis
Strategies for the NCI Disease-oriented in Vitro Antitummor Drug
Screen in Cytotoxic Anti-cancer Drugs: Models and Concepts for Drug
Discovery and Development, Kluwer Academic, Boston, 1992: 11-34; each
of which is hereby incorporated herein in its entirety by express
reference thereto, have been used to estimate the cyto-toxicity of
anti-cancer therapeutics. One of ordinary skill in the art
understands, however, that, although in vitro testing provides a
useful screen for potentially useful compounds, animals such as humans
are sufficiently complex that the actual in vivo cytotoxicity of a
compound is often surprisingly different than that predicted upon the
basis of an in vitro toxicity screen.

U.S. Pat. No. 5,571,520 discloses the use of molecular crystals of
tetrasilver tetroxide, particularly with oxidizing agents to enhance
the efficiency of such devices, for killing pathogenic microorganisms,
such as staph infections. Amounts of 10 ppm sodium persulfate as an
oxidizing agent were used with certain amounts of silver tetroxide in
the reported in vitro testing. One human study involved in vivo curing
of a gynecological yeast infection with 10 ppm of the silver tetroxide
and 40 ppm sodium persulfate. Other in vivo topical studies report in
conclusory fashion the cure of a single case of athlete's foot with a
solution of 100 ppm of the composition and the cure of a single case
of toenail fungus with a 25% suspension of the composition.

U.S. Pat. No. 5,676,977 discloses intravenously injected tetrasilver
tetroxide crystals used for destroying the AIDS virus, AIDS
synergistic pathogens, and immunity suppressing moieties (ISM) in
humans. The crystals were formulated for a single injection at about
40 ppm of human blood. This reference also discloses the compositions
cause hepatomegaly, also known as enlarged liver, albeit with no
reported loss of liver function.

The aforementioned references report detailed descriptions of the
mechanism via which the multivalent silver molecular crystal devices
were believed to operate. The instant inventor also presented a
discussion of such results and concepts at a Seminar entitled
"Incurable Diseases Update" (Weizmann Institute of Science, Rehovot,
Israel, Feb. 11, 1998). The title of this presentation was "Beyond
Antibiotics, Non Toxic Disinfectants and Tetrasil.TM. (Trademark of
applicant for the tetroxide)."

In this paper, it was reported that the effects of the electron
transfer involved with respect to the tetroxide, rendered it a more
powerful germicide than other silver entities. The instant inventor
holds patents for multivalent silver antimicrobials, e.g., U.S. Pat.
No. 5,017,295 for Ag(II) and U.S. Pat. No. 5,223,149 for Ag (III); and
while these entities are stronger antimicrobials than Ag (I)
compounds, they pale by comparison to the tetroxide and so does
colloidal silver that derives its germicidal properties from trace
silver (I) ions it generates in various environments. Accordingly, the
oligodynamic properties of these entities may be summarized as
follows, which is referred to as the Horsfal series:

The other unique property of the tetroxide was that it did not stain
organic matter such as skin in like manner as Ag(I) compounds do. In
addition, it was light stable.

Thus, it is desired to find pharmaceutical compositions and methods
for preventing, treating, or managing one or more cancers or
associated conditions. It is also desired to facilitate the prevention
of future outbreaks of one or more disorders, as well as preventing,
treating, and managing one or more cancerous or related disorder while
avoiding the adverse effects present in many conventional treatments.

SUMMARY OF THE INVENTION

The present invention relates to a method for preventing, treating, or
managing one or more cancerous conditions or dysplastic proliferations
in an animal. The method preferably comprises administering at least
one metal oxide compound or a pharmaceutically acceptable derivative
thereof, to the animal. The metal oxide compound or derivative thereof
preferably comprises a first metal cation having a first valence state
and a second metal cation having a second, different valence state,
such as, for example, an electron active metal oxide compound. The at
least one metal oxide compound or a pharmaceutically acceptable
derivative thereof is preferably administered in an amount and for a
period of time which is therapeutically effective to treat such
condition(s).

In a preferred embodiment, the at least one metal oxide compound or
pharmaceutically acceptable derivative thereof comprises at least one
of Bi(III,V) oxide, Co(II,I) oxide, Cu(I,III) oxide, Fe(II,III) oxide,
Mn(II,III) oxide, Pr(III,IV) oxide, or Ag(I,III) oxide.

The metal oxide compound or derivative thereof is preferably
substantially free of added persulfate.

The invention is preferably adapted to preventing, treating, or
managing systemic cancerous conditions. Preferably, the animal is an
mammal, such as, for example, a human. The metal oxide compound is
preferably administered via intravenous injection or infusion, when
the animal is a human. The intravenous injection or infusion is
preferably subcutaneous, intramuscular, or comprises infusion into the
bloodstream of the animal. Preferably, the administration provides an
amount of the metal oxide sufficient to provide about 1 to about 75
ppm of the metal oxide compound or derivative thereof in the
bloodstream. The metal oxide is preferably administered via infusion
over a period of time sufficient to inhibit adverse side effects, such
as over a time period of from about 30 minutes to about 300 minutes.

The metal oxide compound or derivative thereof may preferably be
administered by a controlled release vehicle. The controlled release
vehicle is preferably implanted in the body at a location suitable for
providing a therapeutically effective amount of metal oxide compound
or derivative thereof to the patient, preferably, without affecting
proper functioning of the animal's liver.

The method of the invention is preferably suitable for cancers or
dysplastic proliferations including at least one of colon cancer, lung
cancer, throat cancer, breast cancer, kidney cancer, pancreatic
cancer, bladder cancer, prostate cancer, uterine cancer, brain cancer,
liver cancer, skin cancer, testicular cancer, stomach cancer, adrenal
gland cancer, cancer of the ovaries, thyroid cancer, bronchial cancer,
tracheal cancer, eye cancer, bone cancer, cervical cancer, oral cavity
cancer, soft tissue cancer, pituitary gland cancer, myeloma, rectal
cancer, esophageal cancer, leukemia, lymphoma, cancerous fibroid
tumors, non-cancerous fibroid tumors, or liver cancer. The method is
preferably suitable for cancers including skin cancer that has
metastasized.

In a preferred embodiment, the metal oxide compound or derivative
thereof is administered in conjunction with at least one other
chemotherapeutic agent. The at least one other chemotherapeutic agent
is preferably administered concurrently with the metal oxide compound
or derivative thereof.

Another embodiment of the invention relates to a method for
preventing, treating, or managing one or more cancerous conditions or
dysplastic proliferations associated with a patient's skin, which
method preferably comprises administering at least one metal oxide
compound or a pharmaceutically acceptable derivative thereof to the
skin in an amount and for a period of time which is therapeutically
effective to treat such cancerous or associated condition(s). The
metal oxide compound or derivative thereof preferably comprises a
first metal cation having a first valence state and a second metal
cation having a second, different valence state.

In a preferred embodiment, the at least one metal oxide compound or
pharmaceutically acceptable derivative thereof comprises at least one
of Bi(III,V) oxide, Co(II,III) oxide, Cu(I,III) oxide, Fe(II,III)
oxide, Mn(II,III) oxide, Pr(III,IV) oxide, or Ag(I,III) oxide.

The metal oxide compound or derivative thereof is preferably
substantially free of added persulfate.

The method of the invention is preferably suitable for preventing,
treating, or managing cancerous conditions or dysplastic
proliferations comprising at least one of dysplastic nevi,
neurofibromatosis, basal cell carcinoma, squamous carcinoma, or
melanoma. The method is preferably suitable for preventing, treating,
or managing conditions comprising symptoms of cancer or conditions
associated with a predisposition to cancer, such as neurofibromatosis.

The administering preferably comprises a carrier medium in which the
at least one metal oxide compound or pharmaceutically acceptable
derivative thereof, is dispersed. Preferably the therapeutically
effective amount of the metal oxide or derivative thereof is from
about 50 ppm to 500,000 ppm, such as from about 400 ppm to about
100,000 ppm, based on the weight of the carrier medium. The carrier
medium may preferably comprise petroleum jelly. The administering of
the composition is preferably topical or transdermal, such as directly
to the skin.

Preferably, the at least one metal oxide compound or pharmaceutically
acceptable derivative thereof, further comprises a thixotropic agent
sufficient to increase adherence of the composition to the skin
without excessive runoff.

The at least one metal oxide compound or pharmaceutically acceptable
derivative thereof may, preferably, be administered in the form of a
powder, such as in the form of metal oxide crystals. The administering
of the powder is preferably topical or transdermal, such as directly
to the skin. Preferably the metal oxide or derivative thereof is
administered at a dosage level of about 10 mg to 500 mg per cm.sup.2
of skin surface. A preferred embodiment of a composition suitable for
application as a powder comprises about 5% metal oxide, such as
tetrasilver tetroxide, and about 95% bismuth subgallate.

Yet another embodiment of the invention relates to a method for
preventing, treating, or managing one or more cancerous conditions
associated with a cervix of a female animal. The method preferably
comprises administering at least one metal oxide compound or a
pharmaceutically acceptable derivative thereof to the cervix in an
amount and for a period of time which is therapeutically effective to
treat such cancerous or associated condition(s). Each metal oxide
compound or derivative thereof preferably comprises a first metal
cation having a first valence state and a second metal cation having a
second, different valence state.

In a preferred embodiment, the at least one metal oxide compound or
pharmaceutically acceptable derivative thereof comprises at least one
of Bi(III,V) oxide, Co(II,III) oxide, Cu(I,III) oxide, Fe(II,III)
oxide, Mn(II,III) oxide, Pr(III,IV) oxide, or Ag(I,III) oxide.

The metal oxide compound or derivative thereof is preferably
substantially free of added persulfate.

The metal oxide compound or derivative thereof are preferably applied
directly to the cervix. The administering preferably comprises a
carrier medium, such as petroleum jelly, in which the at least one
metal oxide compound or pharmaceutically acceptable derivative
thereof, is dispersed, preferably in a therapeutically effective
amount from about 50 ppm to 500,000 ppm, based on the weight of the
carrier medium. The at least one metal oxide compound or
pharmaceutically acceptable derivative thereof is preferably applied
in an amount sufficient to obtain a desired effect and to
substantially inhibit undesirable side effects.

Definitions Section

Suitable definitions are provided herein for some of the terms
relating to the present invention.

The terms "patient" or "subject" as used herein refer to animals,
particularly to mammals. In a preferred embodiment, the terms
"patient" or "subject" refer to humans.

As used herein, the terms "adverse effects," "adverse side effects,"
and "side effects" include, but are not limited to, staining of the
skin, headache, dry mouth, constipation, diarrhea, gastrointestinal
disorders, dry skin, staining of the skin, hepatomegaly, fever,
fatigue, and the like.

The phrase "therapeutically effective amount" when used herein in
connection with the compositions and methods of the invention, means
that amount of metal oxide composition, or a derivative thereof,
which, alone or in combination with other drugs or treatment
modalities, provides a therapeutic benefit in the prevention,
treatment, or management, of one or more of forms of cancer or a
symptom or related condition thereof. Preferably, the therapeutically
effective amount of a component yields the desired therapeutic benefit
without undue adverse side effects (such as toxicity, irritation, or
allergic response) commensurate with a reasonable benefit/risk ratio
when used in the manner of this invention.

The term "substantially free" means less than about 10 weight percent,
preferably less than about 5 weight percent, more preferably less than
about 1 weight percent, and most preferably less than about 0.1 weight
percent of added persulfate is present according to the invention. In
another embodiment, the term "substantially free" refers to the same
amounts of other added oxidizing agents present in the compositions.

The term "controlled-release component" in the context of the present
invention is defined herein as a compound or compounds, including
polymers, polymer matrices, gels, permeable membranes, liposomes,
microspheres, or the like, or a combination thereof, that facilitates
the controlled-release of the active ingredient (e.g., tetrasilver
tetroxide) in the pharmaceutical composition.

The term "about," as used herein, should generally be understood to
refer to both numbers in a range of numerals. Moreover, all numerical
ranges herein should be understood to include each whole integer
within the range.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

It has now been discovered that pharmaceutical compositions comprising
at least one oxide compound or a pharmaceutically acceptable
derivative thereof can be used as advantageous active ingredients in
the prevention, treatment, or management of various cancerous
conditions. The oxide compound preferably comprises a metal oxide,
such as an electron active metal oxide. The metal oxide compound or
pharmaceutically acceptable derivative thereof preferably comprise a
first metal cation having a first valence state and a second metal
cation having a second, different valence state. One of ordinary skill
in the art understands that, in general, the valence state of a
species, such as a metal cation, is related to the charge associated
with or assigned to the species.

Preferably, the at least one metal oxide compound or a
pharmaceutically acceptable derivative comprises at least one electron
active metal oxide compound, such as, for example, at least one of
Bi(III,V) oxide, Co(II,III) oxide, Cu(I,III) oxide, Fe(II,III) oxide,
Mn(II,III) oxide, Pr(II,IV) oxide, or Ag(I,III) oxide. Preferred
compounds of the invention comprise at least one metal tetroxide, such
as silver tetroxide. The terms metal tetroxide and metal tetraoxide,
are synonymous as used herein.

In one preferred embodiment, the metal oxide compound compositions are
substantially free of added persulfate or other added oxidizing
agents, since, when applied topically, such agents may cause adverse
effects, such as skin irritation and skin over-drying. In another
preferred embodiment, the compositions are substantially free of any
oxidizing agents. More particularly, the invention relates to methods
for preventing, treating, and managing cancerous conditions and
conditions associated with cancer.

In one embodiment, the compositions include a molecular scale device
comprising at least one crystal of a metal oxide compound. A plurality
of these metal oxide crystals, such as on the order of trillions, may
be employed in various pharmaceutical formulations and therapies to
effectuate the prevention, treatment, and/or management of various
cancers and conditions associated with cancer. The compositions of the
invention include powders comprising metal oxide crystals of the
invention.

The compositions and methods of the invention advantageously provide a
desired effect such as preventing, treating, or managing cancer or
conditions associated with cancer. "Management," as used herein,
includes controlling one or more cancers, or conditions associated
with such cancer(s), that cannot be cured completely, reducing the
severity of affliction of such cancers or related conditions, and the
like. Thus, a preferred embodiment of the invention relates to a
method of inducing cytotoxicity (cell killing) in cancer cells or
reducing the viability of cancer cells. In one embodiment, the
invention relates to the treatment or management of cancer and/or
diseases or conditions associated with cancer, while in another
embodiment the invention relates to the prevention, of cancer and/or
diseases or conditions associated with cancer.

Preferred metal oxides of the invention comprise a first metal cation
having a first valency state and a second metal cation having a second
valency state, which differs from the first valency, preferably by at
least one charge. The first and second metal cations are preferably
the same metal. Without being bound by theory, it is believed that the
metal oxides of the present invention operate by transferring
electrons between cations of differing valency, the electrons
contributing to the death of the cancer cells by traversing the cell
membrane. By way of non-limiting example, it is believed that the
crystal lattice of a silver tetroxide (Ag.sub.4 O.sub.4) molecular
device operates against cancer, tumors, or cells associated with
cancer by transferring electrons from its two monovalent silver ions
to the two trivalent silver ions in the crystal, contributing to the
death of the cancer cells by traversing their cell membrane surface.
This in effect "electrocutes" the cancer cells. The electrons are
forced out of their balanced crystals by such labile groups as NH,
NH.sub.2, S--S, and SH associated with the cellular surface. Normal
cells are not believed to be affected, because they are not believed
to proliferate fast enough to expose these labile bonds.

The metal oxides of the invention are preferably stable as determined
by the dissociation constants of the compounds. For example, the
dissociation constant (K.sub.A) of Ag.sub.4 O.sub.4 is 7.9.times.
10.sup.-13. Therefore the molecule is not believed to be disturbed
unless more stable complexes are formed with such ligands as those
associated with the cancer cell membrane surface in a dynamic state.
Indeed, the end result of the electron transfer, which is a redox
reaction, is believed to result in the metal ions of a lower valency
being oxidized to a higher valency state and metal ions of a higher
valency state being reduced to a lower valency state.

Returning to the non-limiting example of silver tetroxide, it is
believed that monovalent Ag ions are oxidized to Ag(II) and the
trivalent Ag ions are reduced to the same end product, Ag(II).
Accordingly, the well-known affinity of monovalent silver for certain
elements such as sulfur and nitrogen is believed to be far exceeded
here, for divalent silver is believed to not merely bind to these
elements as does silver, but to actually form chelate complexes with
their ligands. The molecular crystal attraction for the cell membrane
surfaces is thus believed to be driven by powerful covalent bonding
forces.

The electron transfer occurring in the example of silver tetroxide can
be depicted by the following redox half reactions:

It was found by rigorous testing that certain silver tetroxide
containing-compositions were comparatively non- toxic in comparison to
monovalent silver salts. Since these silver tetroxide compositions
were effective at certain ppm concentrations in killing pathogens in
nutrient broth and for water treatment, commercial concentrates were
formulated with 2% of the tetroxide. Prior to the acceptance of the
oxide in commerce, for which EPA registration No. 3432-64 was
obtained, it was necessary for the oxide to undergo a series of
toxicity tests. A 3% concentrate was used and evaluated by a certified
laboratory employing good laboratory practice (GLP) according to the
Code of Federal Regulations for this purpose.

The results were as follows:

Acute Oral Toxicity LD.sub.50 Greater than 5,000 mg/Kg Acute Dermal
Toxicity LD.sub.50 Greater than 2,000 mg/Kg Primary Eye Irritation
Mildly irritating Primary Skin Irritation No irritation Skin
Sensitization Non-Sensitizing

Subsequent evaluations conducted according to the invention showed
that unless persons were prone to silver allergies, the pure tetroxide
compositions according to the invention could be applied to, for
example, the skin without any ill effects or evidence of irritation,
despite the fact that the compositions of the invention can be a
powerful oxidizing agent. This can perhaps be explained by the
stability manifested by the above-noted K.sub.A of the silver
compositions. Accordingly, in a preferred embodiment, the metal oxides
of the invention are applied directly in a powder or composition form
to afflicted areas, such as the skin, cervix, or cervical pelvic
region of an animal afflicted with cancer. Preferred routes of
administration include topically and application to mucosa.
Application can be made, for example, digitally or using a suitable
applicator.

One embodiment of the present invention relates to compositions and
methods of using the metal oxide compositions of the invention while
minimizing the amount of additional oxidizer, such as persulfate. It
has been found in accordance with the present invention that the
additional oxide is not required and in some circumstances is
undesirable when the oxide is applied to, for example, the skin or
cervix, in part due to the undesirable side effect of irritation. In
one embodiment, the compositions are substantially free of added
persulfates, while in a preferred embodiment, the compositions are
completely free of added persulfates. In one preferred embodiment, the
compositions are substantially free of added oxidizer, while in
another preferred embodiment they are completely free of added
oxidizer. The aforementioned compositions may be applied topically or
to mucosa associated with, for example, the skin, cervix, vagina, or
colon.

The metal oxide compound, such as tetrasilver tetroxide, may be black
in color, such that care must be taken when formulating suitable
topical pharmaceutical compositions according to the invention to
inhibit or avoid blackening or staining of the skin. Without being
bound by theory, it is believed that larger amounts of the silver
tetroxide composition promote increased staining. Thus, in one
embodiment, the pharmaceutical compositions preferably have an
insufficient amount of metal oxide compound to cause visible skin
staining.

Where the metal oxide compositions according to the invention are
applied to the skin, they may be combined with a carrier at an amount
from about 5 ppm to 500,000 ppm, more preferably from about 50 ppm to
250,000 ppm of the metal oxide composition, based on the weight of the
carrier. In various embodiments, the compositions are provided in
amounts from about 400 ppm to 100,000 ppm, from about 1,000 ppm to
70,000 ppm, from about 10,000 ppm to 50,000 ppm, or from about 20,000
ppm to 40,000 ppm. In one preferred embodiment, the compositions are
formulated with about 25,000 ppm to 35,000 ppm of metal oxide. It will
be readily understood by those of ordinary skill in the art that the
administration of 0.005 g of metal oxide to an adult human being
provides about 1 ppm of the metal oxide in the bloodstream of the
human. In another embodiment, the concentration of the metal oxide
crystals dispersed in the carrier ranges from about 0.1 to 10% by
weight, more preferably from about 0.25 to 5% by weight and most
preferably from about 2 to 4% by weight. The compositions, when
applied topically, can be applied to the skin about 1 to 3 times per
day until the condition is suitably cured or satisfactorily
controlled. In one embodiment, the composition may generally be
topically applied at a dosage level of from about 1 mg to 1000 mg per
cm.sup.2 of skin surface, preferably about 10 mg to 500 mg per cm.sup.
2 of skin surface.

A preferred carrier for topical formulations and administration
includes petroleum jelly, such as white petroleum jelly. For example,
a suitable white petroleum jelly is available from Penreco of Houston,
Tex.

A preferred mode of application of the oxide of the invention is as an
ointment. Suitable formulations include, but are not limited to,
salves and the like. If desired, these may be sterilized or mixed with
auxiliary agents, e.g., thixotropes, stabilizers, wetting agents, and
the like. Preferred vehicles include ointment bases, e.g.,
polyethylene glycol-1000 (PEG-1000); conventional ophthalmic vehicles;
creams; and gels, as well as petroleum jelly and the like.

The cancerous conditions and diseases that may be prevented, treated,
or managed with the compositions of the invention vary and include,
but are not limited to, cancers including any of the various malignant
neoplasms, tumors, or cells, such as, for example, those marked by a
proliferation of anaplastic cells. In particular, the term cancer
includes any cancers that involve specific organs or regions of the
body such as the colon, lung, throat, breast, kidney, pancreas,
bladder, prostate, uterus, brain, liver, skin, testicles, stomach,
adrenal gland, ovaries, thyroid, rectum, bronchus, trachea, eye, bone,
cervix, oral cavity, soft tissue, pituitary gland, myeloma, rectum,
esophagus or liver. The invention is also suited for the prevention,
treatment, or management of cancerous fibroid tumors and non-cancerous
fibroid tumors. Prevention, treatment, or management of any of the
above conditions, as well as any others described herein, individually
or in any combination, simultaneously or concurrently, is contemplated
according to the invention.

Also included are various cell proliferations such as leukemia, which
is a malignant overproduction of white blood cells, lymphoma, and
metastasized melanoma which has proliferated from skin via blood and/
or the lymphatic system. Conditions or diseases associated with a
predisposition to cancer, such as, for example, neurofibromatosis are
also included. The present invention preferably allows treatment or
management of conditions or diseases associated with a predisposition
to cancer even if those conditions or diseases have not fully
progressed to a cancerous or malignant stage.

The present invention is also adapted to treating or managing atypical
proliferations of cells, such as those characterized by nuclear
enlargement and failure of maturation and differentiation. Such
proliferations may be short of malignancy. Atypical proliferations
suitable for treatment, management, or prevention by the present
invention include dysplasia or dysplastic proliferations, such as
dysplastic nevi or neurofibromatosis, which are recognized by
alterations in the appearance of cells (cytology). Dysplastic cells
may have some of the features of malignant cells but the changes are
less pronounced. As the dysplasia progresses, the nuclei of cells
become more hyperchromatic and the nuclear membranes become more
irregular; the size of the nucleus increases and the cytoplasm does
not increase proportionately, so the that the nuclear:cytoplasmic
ratio increases.

Different therapeutically effective amounts and deliver systems may be
applicable for each disorder, as will be readily known or determined
by those of ordinary skill in the art.

Tumors or neoplasms include new growths of tissue in which the
multiplication of cells is uncontrolled and progressive. Some such
growths are benign, but others are termed "malignant," leading to
death of the organism. Malignant neoplasms or "cancers" are
distinguished from benign growths in that, in addition to exhibiting
aggressive cellular proliferation, they invade surrounding tissues and
metastasize. Moreover, malignant neoplasms are characterized in that
they show a greater loss of differentiation (greater
"dedifferentiation"), and a greater loss of their organization
relative to one another and their surrounding tissues. This property
is also called "anaplasia."

Neoplasms preventable, treatable, or manageable by the present
invention include all solid tumors, i.e., carcinomas and sarcomas.
Carcinomas include those malignant neoplasms derived from epithelial
cells which tend to infiltrate (invade) the surrounding tissues and
give rise to metastases. Adenocarcinomas are carcinomas derived from
glandular tissue or in which the tumor cells form recognizable
glandular structures. Sarcomas broadly include tumors whose cells are
embedded in a fibrillar or homogeneous substance like embryonic
connective tissue.

The invention can also be practiced by administering the metal oxide
compositions in conjunction with one or more other anti-cancer
compatible chemotherapeutic agents, such as any conventional
chemotherapeutic agent. The combination of the metal oxide with such
other agents can potentiate the chemotherapeutic protocol. Numerous
chemotherapeutic protocols will present themselves in the mind of the
ordinary-skilled practitioner as being capable of use according to the
methods of the invention. Any compatible chemotherapeutic agent can be
used, including antimetabolites, hormones and antagonists,
radioisotopes, as well as natural products. For example, the metal
oxide can be administered with taxol and its natural and synthetic
derivatives, and the like, and combinations thereof. As another
example, in the case of mixed tumors, such as adenocarcinomas of the
breast and prostate, in which the tumors can include gonadotropin-
dependent and gonadotropin-independent cells, the metal oxide can be
administered in conjunction with leuprolide or goserelin (synthetic
peptide analogs of LH-RH), or both. Other antineoplastic protocols
include the use of a metal oxide with another treatment modality,
e.g., surgery, radiation, other chemotherapeutic agent, etc., referred
to herein as "adjunct antineoplastic modalities." Thus, the method of
the invention can be employed with such conventional regimens with the
benefit of reducing side effects and enhancing efficacy. These other
anti-cancer chemotherapeutic agents and modalities may be administered
either concurrently or sequentially with the metal oxide compositions
of the invention.

A preferred metal oxide for use according to the invention,
tetrasilver tetroxide, has been commercially sold under the poorly
named "Ag(II) OXIDE" tradename. It may also be obtained from Aldrich
Chemical Co., Milwaukee, Wis. The chemical synthesis of silver oxide
compounds according to the invention can be performed according to the
method described on page 148 in M. Antelman, "Anti-Pathogenic
Multivalent Silver Molecular Semiconductors," Precious Metals, vol.
16:141-149 (1992) by reacting silver nitrate with potassium
peroxydisulfate according to the following equation in alkali
solutions:

The magnitude of a prophylactic or therapeutic dose of metal oxide
composition(s), or a derivative thereof, in the acute or chronic
management of diseases and disorders described herein will vary with
the severity of the condition to be prevented, treated, or managed and
the route of administration. For example, oral, mucosal (including
vaginal and rectal), parenteral (including subcutaneous,
intramuscular, bolus injection, and intravenous, such as by infusion),
sublingual, transdermal, nasal, buccal, and like may be employed.
Dosage forms include tablets, troches, lozenges, dispersions,
suspensions, suppositories, solutions, capsules, soft elastic gelatin
capsules, patches, and the like. The dose, and perhaps the dose
frequency, will also vary according to the age, body weight, and
response of the individual patient. Suitable dosing regimens can be
readily selected by those of ordinary skill in the art with due
consideration of such factors.

In general, for topical and mucosal application, such as application
to the skin or cervix, the total daily dosage for the conditions
described herein can be from about 1 mg to 500 mg of the metal oxide
or derivative thereof, while in another embodiment, the daily dosage
can be from about 2 mg to 200 mg of the metal oxide composition. A
unit dosage can include, for example, 30 mg, 60 mg, 90 mg, 120 mg, or
200 mg of metal oxide composition. Preferably, the active ingredient
is administered in single or divided doses from one to four times a
day.

In another embodiment, the compositions are administered by an oral
route of administration. The oral dosage forms may be conveniently
presented in unit dosage forms and prepared by any methods available
to those of ordinary skill in the art of pharmacy.

In managing the patient, the therapy may be initiated at a lower dose,
e.g., from about 1 mg, and increased up to the recommended daily dose
or higher depending on the patient's global response. It is further
recommended that children, patients over 65 years, and those with
impaired renal or hepatic function, initially receive low doses when
administered systemically, and that they be titrated based on
individual response(s) and blood level(s). It may be necessary to use
dosages outside these ranges in some cases, as will be apparent to
those of ordinary skill in the art. Furthermore, it is noted that the
clinician or treating physician will know how and when to interrupt,
adjust, or terminate therapy in conjunction with individual patient
response.

Any suitable route of administration may be employed for providing the
patient with an effective dosage of metal oxide, or a pharmaceutically
acceptable derivative thereof. The most suitable route in any given
case will depend on the nature and severity of the condition being
prevented, treated, or managed. One preferred route is parenterally,
preferably intravenously. In this embodiment, a preferred intravenous
route of administration is by infusion.

In practical use, metal oxide, or a derivative thereof, can be
combined as the active ingredient in intimate admixture with a
pharmaceutical carrier according to conventional pharmaceutical
compounding techniques. The carrier may take a wide variety of forms
and may include a number of components depending on the form of
preparation desired for administration. The compositions of the
present invention include, but are not limited to, suspensions,
solutions and elixirs; aerosols; or carriers, including, but not
limited to, starches, sugars, microcrystalline cellulose, diluents,
granulating agents, lubricants, binders, disintegrating agents, and
the like.

Another suitable route of administration of the silver tetroxide
compositions of the invention is topically, e.g., either directly as a
powder or in non-sprayable or sprayable form. Topical administration
is a preferred route of administration for treating topical cancerous
conditions, such as skin cancer that has not metastasized or cervical
cancer. In one embodiment, the metal oxide may be applied topically to
the affected skin areas directly in powder form or in compounded
formulations.

Non-sprayable forms can be semi-solid or solid forms including a
carrier indigenous to topical application and preferably having a
dynamic viscosity greater than that of water. Suitable formulations
include, but are not limited to, suspensions, emulsions, creams,
ointments, powders, liniments, salves and the like. If desired, these
may be sterilized or mixed with any available auxiliary agents,
carriers, or excipients, e.g., thixotropes, stabilizers, wetting
agents, and the like. One or more thixotropic agents can be included
in types and amounts sufficient to increase adhesion of topically
applied compositions of the invention to a surface or mucosa
associated with a treatment zone such as, for example, the skin,
vagina, or cervix, so as to inhibit or prevent runoff or other loss of
the composition from the treatment zone, particularly when the
compositions are formulated for topical administration. With respect
to conditions associated with the skin, the compositions preferably
prevent, treat, or manage such conditions or diseases without visibly
staining the skin, i.e., no staining to the naked eye.

Preferred vehicles for non-sprayable topical preparations include
ointment bases, e.g., polyethylene glycol-1000 (PEG-1000);
conventional ophthalmic vehicles; creams; and gels, as well as
petroleum jelly and the like. In one preferred topical embodiment, the
carrier includes a petroleum jelly. In another preferred topical
embodiment, the carrier is formulated as a cream, gel, or lotion. A
preferred composition comprises about 3% metal oxide, such as
tetrasilver tetroxide, about 47% white petrolatum, about 36% heavy
mineral oil, and about 14% TIVAWAX P Tivian Laboratories Inc.,
Providence, R.I. These topical preparations may also contain
emollients, perfumes and/or pigments to enhance their acceptability
for various uses.

In a preferred embodiment, a metal oxide, or a derivative thereof, is
formulated for parenteral administration by injection (subcutaneous,
bolus injection, intramuscular, or intravenous, such as by infusion),
and may be dispensed in a unit dosage form, such as a multidose
container or an ampule. Parenteral administration is a preferred
administration route when the cancer is systemic, i.e., has a locus
inside the body. Preferably, the formulation adapted for parenteral
administration includes an insufficient amount of persulfate to induce
irritation or adverse side effects. In one preferred embodiment, the
formulation is substantially free of added persulfate, while in
another more preferred embodiment, the formulation is completely free
of added persulfate.

When administered intravenously, such as by infusion, the dosage
preferably provides a concentration of the metal oxide in the blood
stream of about 1 ppm to about 75 ppm, more preferably from about 5
ppm to about 50 ppm, such as from about 10 ppm to about 40 ppm or
about 50 to 200 mg. In a preferred embodiment, a one-time dosage is
infused or injected directly into the bloodstream.

The intravenous dosage is preferably delivered over a period of time
sufficient to substantially inhibit or even avoid the occurrence of
side effects. For example, the dosage can be delivered by
intravenously or by infusion over a time from about 10 minutes to
about 300 minutes, preferably from about 20 minutes to about 240
minutes.

Compositions of the metal oxide, or a pharmaceutically acceptable
derivative thereof, for parenteral administration may be in the form
of suspensions, solutions, emulsions, or the like, in aqueous or oily
vehicles, and in addition to the active ingredient may contain one or
more formulary agents, such as dispersing agents, suspending agents,
stabilizing agents, preservatives, and the like.

Pharmaceutical compositions of the present invention may be orally
administered in discrete pharmaceutical unit dosage forms, such as
capsules, cachets, soft elastic gelatin capsules, tablets, or aerosols
sprays, each containing a predetermined amount of the active
ingredient, as a powder or granules, or as a solution or a suspension
in an aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion,
or a water-in-oil liquid emulsion. Such compositions may be prepared
by any of the methods of pharmacy, but all methods include the step of
bringing into association the active ingredient with the
pharmaceutically acceptable carrier which constitutes one or more
necessary ingredients. In general, the compositions are prepared by
uniformly and intimately admixing the active ingredient with liquid
carriers or finely divided solid carriers or both, and then, if
necessary, shaping the product into the desired presentation. Suitable
types of oral administration include oral solid preparations, such as
capsules or tablets, or oral liquid preparations. If desired, tablets
may be coated by standard aqueous or non-aqueous techniques.

For example, a tablet may be prepared by compression or molding,
optionally, with one or more accessory ingredients. Compressed tablets
may be prepared by compressing in a suitable machine the active
ingredient in a free-flowing form such as powder or granules,
optionally mixed with a binder, lubricant, inert diluent, granulating
agent, surface active agent, dispersing agent, or the like. Molded
tablets may be made by molding, in a suitable machine, a mixture of
the powdered compound moistened with an inert liquid diluent. In one
embodiment, each tablet, capsule, cachet, or gel cap contains from
about 0.5 mg to about 500 mg of the active ingredient, while in
another embodiment, each tablet contains from about 1 mg to about 250
mg of the active ingredient. The amount of active ingredient found in
the composition, however, may vary depending on the amount of active
ingredient to be administered to the patient.

Another suitable route of administration is transdermal delivery, for
example, via an abdominal skin patch.

The metal oxide, or a suitable derivative thereof, may be formulated
as a pharmaceutical composition in a soft elastic gelatin capsule unit
dosage form by using conventional methods well known in the art, such
as in Ebert, Pharm. Tech, 1(5):44-50 (1977). Soft elastic gelatin
capsules have a soft, globular gelatin shell somewhat thicker than
that of hard gelatin capsules, wherein a gelatin is plasticized by the
addition of plasticizing agent, e.g., glycerin, sorbitol, or a similar
polyol. The hardness of the capsule shell may be changed by varying
the type of gelatin used and the amounts of plasticizer and water. The
soft gelatin shells may contain a preservative, such as methyl- and
propylparabens and sorbic acid, to prevent the growth of fungi. The
active ingredient may be dissolved or suspended in a liquid vehicle or
carrier, such as vegetable or mineral oils, triglycerides, surfactants
such as polysorbates, or a combination thereof.

In the case of tumors having loci inside the body, e.g., brain tumors,
prostate tumors, and the like, the metal oxide can be delivered via a
controlled release delivery vehicle. In a preferred embodiment, the
controlled release vehicle includes a polymeric material, delivered or
surgically implanted at or near the lesion site. One of ordinary skill
in the art will be familiar with controlled release means and delivery
devices, such as those described in U.S. Pat. Nos.: 3,845,770;
3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533; 5,059,595;
5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; and 5,733,566,
the disclosures of which are hereby incorporated herein by express
reference thereto. These pharmaceutical compositions can be used to
provide slow or controlled-release of the active ingredient therein
using, for example, hydropropylmethyl cellulose in varying proportions
to provide the desired release profile, other polymer matrices, gels,
permeable membranes, osmotic systems, multilayer coatings,
microparticles, liposomes, microspheres, or the like, or a combination
thereof. Suitable controlled-release formulations available to those
of ordinary skill in the art, including those described herein, may be
readily selected for use with the metal oxide compositions of the
invention. Thus, single unit dosage forms suitable for topical,
parenteral, or oral administration, such as infusions, intravenous
drips, gels, lotions, cremes, tablets, capsules, gelcaps, caplets, and
the like, that are adapted for controlled-release are encompassed by
the present invention.

All controlled-release pharmaceutical products have a common goal of
improving drug therapy over that achieved by their non-controlled
counterparts. Ideally, the use of an optimally designed controlled-
release preparation in medical treatment is characterized by a minimum
of drug substance being employed to cure or control the condition in a
minimum amount of time. Advantages of controlled-release formulations
may include: 1) extended activity of the drug; 2) reduced dosage
frequency; and 3) increased patient compliance.

Most controlled-release formulations are designed to initially release
an amount of drug that promptly produces the desired therapeutic
effect, and gradual and continual release of other amounts of drug to
maintain this level of therapeutic effect over an extended period of
time. In order to maintain this constant level of drug in the body,
the drug should be released from the dosage form at a rate that will
replace the amount of drug being metabolized and excreted from the
body.

The controlled-release of the active ingredient may be stimulated by
various inducers, for example pH, temperature, enzymes, water, or
other physiological conditions or compounds. The pharmaceutical
compositions for use in the present invention include the metal oxide,
or a derivative thereof, as the active ingredient, and may also
contain a pharmaceutically acceptable carrier, and optionally, other
therapeutic ingredients. Suitable derivatives include any available
"pharmaceutically acceptable salts," which refer to a salt prepared
from pharmaceutically acceptable non-toxic acids including inorganic
acids, organic acids, solvates, hydrates, or clathrates thereof.
Preferably, in the case of silver (I,III), the salts do not comprise
halides. Examples of such inorganic acids are hydrochloric,
hydrobromic, hydroiodic, nitric, sulfuric, and phosphoric. Appropriate
organic acids may be selected, for example, from aliphatic, aromatic,
carboxylic and sulfonic classes of organic acids, examples of which
are formic, acetic, propionic, succinic, citric, fumaric, gluconic,
isethionic, lactic, malic, mucic, tartaric, para-toluenesulfonic,
glycolic, glucuronic, maleic, furoic, glutamic, salicylic, mandelic,
methanesulfonic, ethanesulfonic, benzenesulfonic (besylate),
sulfanilic, alginic, galacturonic, and the like. Particularly
preferred acids phosphoric, methanesulfonic, and glycolic.

EXAMPLES

These and other aspects of the present invention may be more fully
understood with reference to the following non-limiting examples,
which are merely illustrative of the preferred embodiment of the
present invention, and are not to be construed as limiting the
invention, the scope of which is defined by the appended claims.

Example 1

Treatment of Breast Cancer According to the Invention

A subject group was formed of thirty female residents of Central
America aged 32 to 52 years who had been diagnosed as having breast
cancer. The subjects had the following general characteristics: 38%
were older than 45 years old; 47% had never borne children; 60% were
around menopause;18% had their first period before age 13 years; 60%
of had used oral contraceptives; 38% of the subjects had cancers
associated with the left breast; and 40% of the subjects had a family
history of breast cancer. The diagnosis of each subject was confirmed
by biopsy and a mammogram was acquired from each subject.

Each subject was evaluated daily by an oncologist(s) and each received
a single dosage of tetrasilver tetroxide by IV, i.e., intravenously,
sufficient to provide a concentration in the bloodstream of 10 ppm.
One-half of the patients received the dosage in 10 minutes by IV
injection. The other half of the subjects received the dosage by IV
injection over a 4-hour period.

As discussed below, the subjects were arranged in three histology
groups. Within each histologic group, 50% of the subjects received the
dosage via the 10 minute IV and the other 50% of the subjects received
the dosage over the 4 hour period of time.

Prior to initiating treatment, the tetrasilver tetroxide was subjected
a quality assurance protocol to reduce side effects.

Group I: Group I included 10 patients who had been diagnosed as having
infiltrative canalicular breast carcinoma.

A 1 cm diameter biopsy fragment was sent to a pathologist laboratory.
The pathologist reported an increase of the dense fibrous tissue,
anaphasic cells in the gland ducts, forming lines, tubes, ducts,
glands and cell anastomosis. The histologic report confirmed the
diagnosis of the 10 patients in this group: infiltrative canalicular
breast carcinoma.

Group II: Group II included 10 subjects were diagnosed as having
ductile carcinoma special type, medular breast cancer.

Group III: Group III included ten patients diagnosed as having
infiltrative lobular breast cancer.

For each of the three groups, the dosage was 10 ppm for every subject.
The patients had 24-hour evaluations by three oncologists who were
each responsible for an 8-hour time period. Every 4 hours, a
professional nurse acquired vital signs from the subjects. Twenty-four
hour hemodynamic monitoring was performed. The subjects walked 2 hours
after they received the dosage and they were prescribed a free diet.
Every twenty-four hours, urine was collected. Every seven days, the
following laboratory tests were performed: a complete blood count,
hemoglobin, hematocracity, vcm, vhcm, complete red blood cells,
complete white blood cells, albumin, bilirubin, calcium, and
cholesterol, creatine, glucose, ldh, potassium, sodium, triglyceride,
uric acid, urea nitrogen, AST and SGOT.

Results of IV Tetrasilver Tetroxide Injection

Group I: Forty-eight hours after treatment, the texture of the nodules
had changed from hard as a stone to a mild, soft nodule, the redness
was almost gone, and the retraction of the nipple was the same. At 12
days after treatment, the redness was gone, the nodule was no longer
discernable by touch, and the nipple retraction had disappeared. Three
of the patients who had received 10 minute IV injection exhibited an
increase in body temperature and a slight liver enlargement. Liver
function was not affected, however, as demonstrated by normal liver
function tests, a normal red blood cell count, and a normal white
blood cell count. An electrocardiogram did not reveal abnormalities.
Sodium, potassium, and magnesium blood levels decreased. Albumin,
bilirubin calcium, cholesterol, creatinine, LDH, AST, SGOT, and
triglyceride remained normal.

At 19 days after the treatment, the oncologist ordered a new biopsy of
this group. The pathologist reported 100% of the biopsies as
intralobular ducts normal in number and size. No change in shapes.
Cylindric cells in the mammary ducts were normal in shape and in size.
Basal membranes were intact. Diagnosis: normal mammary tissue.

After 21 days of treatment, the patients were again evaluated and all
of them had tissue retractions in the area where the nodule was
located. The subjects had no more symptoms. The patients were allowed
to return home after 25 days and given a return appointment in 30
days. The final biopsy revealed no difference between the group that
received direct injection and the group that received IV solution. A
little quicker response in the IV solution patients was observed in
comparison with the direct injection group.

Group II: At 36 hours after treatment, the formerly 10 cm on average
tumors had decreased to an average of 8.4 cm and the lymph nodes were
smaller and the texture was changed. At day number 16 after treatment,
the huge tumor mass was gone, the nodule was no longer palpable, and
the lymph enlargement had disappeared. One patient of the sub-group
that received direct IV injection exhibited an increase in body
temperature and a slight liver enlargement. Liver function was not
affected, however, as demonstrated by normal liver function tests, a
normal red blood cell count, and a normal white blood cell count. An
electrocardiogram did not reveal abnormalities. Sodium, potassium, and
magnesium blood levels decreased. Albumin, bilirubin calcium,
cholesterol, creatinine, LDH, AST, SGOT, and triglyceride remained
normal.

At day number 23 after the treatment, the oncologist ordered a new
biopsy of Group II. The biopsy was acquired after 24 days of
treatment. The oncologist reported that there were no more
hemorrhaging and necrosis zones in the breast tissue. The pathologist
reported 90% of the biopsy as intralobular ducts normal in number and
size. No change in shape and normal mitosis was observed. Cilindric
cells in the mammary ducts were normal in shapes and in sizes and
basal membranes were intact. Diagnosis: normal mammary tissue.

Two days later, the patients were evaluated and all of them had tissue
retractions in the area where the nodule was located. No more symptoms
were exhibited. Six days later, the doctor allowed them to return home
with a follow-up appointment scheduled for 30 days. The final biopsy
result did not present a difference between the group that received
direct injection and the group that received IV solution. A slightly
quicker response was observed in the IV solution patients in
comparison with the direct injection group.

Group III: At 5 days after treatment, the nipple retraction, bleeding
from the nipple, the distorted areola, and the attachment of the mass
to surrounding tissues were almost gone. The average nodule size
decreased from 4 cm to 2.6 cm. At day number 20 after treatment, the
retracted nipples, the bleeding from the nipple, the distorted areola
and the attachment of the mass to surrounding tissues was in 90%
remission. The average size of the nodules had decreased further from
2.6 cm to 1 cm.

None of the patients of this group that had received direct injection
exhibited an increase in the body temperature or even slight liver
enlargement. The red blood cells count was normal, and the white blood
cells remained normal. The electrocardiogram did not reveal
abnormalities. Sodium, potassium, and magnesium blood levels deceased.
Albumin, bilirubin calcium, cholesterol, creatinine, LDH, AST, SGOT
and triglyceride remained normal.

At day number 29 after the treatment, the oncologist ordered a new
biopsy of this group. The pathologist reported 80% of the biopsies as
intralobular ducts normal in number and size. No change in shape.
Cilindric cells in the mammary ducts normal in shape and in size and
the basal membranes were intact. Diagnosis: normal mammary tissue.

Seven days later, the patients were evaluated and all of them had
tissue retractions in the area where the nodule was located and did
not exhibit any more symptoms. One day later, the patients were
allowed to return home and given a follow up appointment in 30 days.
The final biopsy result did not indicate a difference between the
group that received direct injection and the group that received IV
solution. A slightly quicker response was observed, however, for the
IV solution patients in comparison with the direct injection group.

Conclusions of IV Tetrasilver Tetroxide Study

1. Tetrasilver tetroxide is preferably delivered in an IV solution to
inhibit undesirable side effects.

2. Tetrasilver tetroxide administered by IV appears to stop the growth
of the breast cancer.

3. Tetrasilver tetroxide appears to stimulate the normal breast cells
and allows them to replace the anaphasic cells in the breast
carcinoma.

4. Tetrasilver tetroxide appears to cure infiltrative breast carcinoma
in a 24 day period.

5. Tetrasilver tetroxide appears to cure ductile carcinoma special
type, medular breast carcinoma in a 30 day period.

6. Tetrasilver tetroxide appears to cure infiltrative lobular breast
cancer in a 30 days period.

7. Although certain patients developed mild cases of hepatomegaly, the
liver functioning was not impaired as evidenced by the normal levels
of liver function enzymes in the blood stream.

Example 2

Treatment of Paget's Disease of the Nipple

A study was performed to determine the affect of the tetrasilver
tetroxide compositions of the invention on patients suffering from
Paget's disease of the nipple. The compositions were applied in
ointment form in this study. Paget's disease of the nipple is a rare
type of carcinoma that appears as a unilateral dermatitis of the
nipple and represents extension to the epidermis of an underlying
mammary duct carcinoma. The redness, oozing, and crusting closely
resemble dermatitis, but the physician should suspect carcinoma
because the lesion is unilateral.

This study were performed in 25 patients, between 42 to 59 years old.
Each patient suffered from Paget's of the nipple, a diagnosis
confirmed by biopsy. As discussed below, each patient was placed in
one of two groups.

Group 1: Group I included 13 patients with Paget's disease of the
nipple, with an average depth of invasion of 0.76 to 1.5 mm. All of
the laboratory results indicated the same diagnosis and were taken
weekly for 4 weeks. Each patient in this group exhibited metastasis.
The lesions were all ulcered and sized on average from 2.3 cm to 3.4
cm. The treatment protocol began with 200 mg of ointment containing 3%
tetrasilver tetroxide 3 times per day as applied by a physician. All
of the patients were evaluated daily for 4 weeks. None of the patients
received anterior treatment.

Group II: Group II included 12 patients suffering from Paget's disease
of the nipple, with an average depth of invasion of 2.26 cm to 3.0 cm.
All the laboratory results for 4 weeks confirmed the diagnosis. The
lesions were all ulcered and sized from an average of 4.3 cm to 5.2
cm. The treatment protocol began with 200 mg of ointment containing 3%
tetrasilver tetroxide 3 times per day as applied by a physician. All
of the patients were evaluated daily for 4 weeks. None of the patients
had anterior treatment.

Results of Paget's Disease Treatment

Group I: Over a period of 15 days, the lesions from Paget's disease
began to regress and dry out in all the patients. Pain was gone and
the patients began to regain an appetite. The color of the lesions
changed from a red more to white. At day 27 of the study, all of the
lesions were healed and the lesions were not visible at all. No
recurrences of lesions were observed. The last biopsy showed minimum
amount of atypical cells, and the mammary ducts were normal.

Group II: Over a period of 23 days the lesions began to regress and
became dryer. By the 29.sup.th day of the study, it was almost
impossible to distinguish the lesions. All the patients began to eat
normally again and the last biopsy revealed 18% of atypical cells. No
recurrences of lesions were found. None of the patients had exhibited
side effects.

Conclusions of Paget's Disease Study

The final biopsies, however, revealed 18% atypical cells, suggesting
that additional time is required for the tetrasilver tetroxide to
completely eliminate the presence of atypical cells.

Example 3

Treatment of Rhabdomyosarcoma with Tetrasilver TetroxideOintment 3%

A study group was formed of twelve patients aged between 45 and 65
years. Each patient had been diagnosed with ulcerative
Rhabdomyosarcoma. A pathologist confirmed the diagnosis of each
patient based upon a biopsy. All the patients were Caucasian and had
similar exposures to sunlight during their lifetimes. All patients had
infections demonstrating the presence of pathogens received during
chemotherapy and surgical resection treatment. None of the patients
exhibited signs of metastasis. As discussed below, the patients were
divided into two groups.

Group I: Group I was formed of seven patients with confirmed biopsy
diagnosis of ulcerative Rhabdomyosarcoma with extreme infection. Each
patient exhibited ulcerative injuries at the inferior member with an
extension of 9 cm to 12 cm in diameter. The ulcers exhibited
serosanguinous secretions in abundance. The evaluation period began
from 4 to 7 months prior to initiating treatment with 3% tetrasilver
tetroxide. Each patient started treatment with 200 mg of ointment 3%,
three times a day for 30 days.

Group II: Group II was formed of five patients with confirmed biopsy
diagnosis of ulcerative Rhabdomyosarcoma with extreme infection. Each
case presented extensive ulcerative injuries with indurated edges
larger than 12 cm in diameter. The period of evaluation was more than
7 months and all the patients received chemotherapy and surgical
resection. Each patient was treated with ointment of 3% of tetrasilver
tetroxide of the invention three times a day for thirty days.

Results

Group I: All patients experienced a commencement of healing of the
ulcers by day 27 from the start of the treatment. The regions of
irritation receded and the color of the lesions became darker and more
similar to normal skin color. By day 30, the ulcers were dry and began
the scarring process. A biopsy control preformed at the 30.sup.th day
revealed normal muscular tissue with no signs of metaplasia. At the
40.sup.th day of treatment, a biopsy control was preformed and
indicated that the injured metaplastic cells had been replaced by
cells of normal appearance. By the 45.sup.th day, the ulcers were no
longer visible. The post-treatment evaluations showed no signs of
recidivism.

Group II: All patients showed a commencement of the healing of the
ulcers by day 28 from the start of the treatment. By day 40, the
ulcers had almost disappeared and a biopsy confirmed that 80% of the
diseased tissue had been replaced with healthy tissue. From a clinical
standpoint, the ulcers were no longer visible.

The tests demonstrated that the topical application of tetrasilver
tetroxide was effective in curing infections and healing skin ulcers
associated with Rhabdomyosarcoma, and without significant adverse
effects.

Example 4

Topical Treatment with Tetrasilver Tetroxide on Neurofibromatosis

This study was performed to determine the effect of topical treatment
with tetrasilver tetroxide on neurofibromatosis. A study group was
formed of twelve patients aged 5 months to 3 years who had been
diagnosed as having neurofibromatosis. These diagnoses were
reconfirmed in conjunction with this study. Diagnoses were made by
clinical study and by biopsy of the lesions. None of the patients had
received prior treatment. None of the patients exhibited skeletal
anomalies or lesions of the optic nerve or acoustic nerve. The
patients were arranged in 2 groups as discussed below.

Group I: Group I included eight patients having von Recklinghausen's
disease and neurofibromatosis type plexiform neuromas. All of them
were symptomatic. The brown spots of the skin were located in the
trunk. Each patient applied 200 mg of ointment with 3% tetrasilver
tetroxide, 3 times per day for 30 days. Daily evaluations were made to
observe progress and to determine the presence of side effects.

Group II: Group II included four patients who had been diagnosed as
having von Recklinohausen's disease and neurofibromatosis type
neurofibroma. Each diagnosis was confirmed by biopsy. All of the
patients were symptomatic. The lesions were located in the trunk,
pelvis, and elbows. All of the patients of this group applied 200 mg
of ointment with 3% of tetrasilver tetroxide 2 times per day for 30
days. Daily evaluations were made to observe progress and determine
the presence of side effects.

Results of Neurofibromatosis Study

Group I: By day 20, five of the eight patients were cured of the spot-
type skin lesions. No more symptoms were found. The biopsy result
showed normal cells and no reoccurrences were observed. The biopsy
results of the other 3 patients showed some atypical cells and the
spots, although reduced in color, were still visible.

Group II: At the end of the study, these patients were still
symptomatic and the biopsies confirmed the presence of atypical
Schwann tumor cells.

Conclusions of Neurofibromatosis Study

Tetrasilver tetroxide ointment seems to have had a positive effect in
neurofibromatosis of the plexiform neuromas type. Higher dosages
produced a better effect for treatment and management of plexiform
neurofibromas.

Tetrasilver tetroxide ointment did not generally seem to produce as
thorough a curative result for neurofibroma.

Example 5

Topical Treatment with Tetrasilver Tetroxide on Cervical Carcinoma

In the following examples, the cervical cancer afflictions are divided
into two main categories. The first is based on cervical smear (PAP
smear) test results following the Bethesda System for reporting
cervical cytologic diagnoses bearing designations CIN-- the acronym
for cervical intraepithelial neoplasia. The second is based on
designated NIC stages of which: 0=carcinoma in situ, intraepithelial
carcinoma; 1=carcinoma strictly confined to the cervix; and 1A
indicates microinvasive carcinoma.

Exetec Lab S.A. located in Honduras, Central America, which performs
clinical tests for major pharmaceutical companies did the clinical
evaluations of tetrasilver tetroxide on the patients having various
cervical cancers. All the clinical testing involved applying 300 mg of
an ointment comprising 3% tetrasilver tetroxide dispersed in a
hydrocarbon base comprising mostly white petrolatum and mineral oil
once a day to the affected cervical/pelvic area.

Five patients classified with CIN 1 cervical cancer, according to
cytologic diagnoses were selected. The diagnoses were conducted two
months prior to the tetrasilver tetroxide clinical trials. The
ointment was applied directly to the cervix and its entrance
(endocervix and exocervix) by a skilled physician. The period of
administration was tend days per patient. Evaluations were made for
any side effects. A biopsy was taken at the end of the treatment. The
biopsies indicated that all of the patients were cured without any
recurrences.

Five patients who were confirmed as CIN 2, i.e., having high grade
squamous intraepithelial lesions including moderate dysplasia 3 months
prior to the clinical studies. Two of the patients had familial
history of the cervical cancers. This group was treated as outlined by
the protocol shown above for 10 days. All patients were cured. The
cytologist reported no more atypical cells present and the
cytopathologist reported normal cells with no recurrences.

Five patients were confirmed as CIN 3, i.e., having high grade
squamous intraepithelial lesions, severe dysplasia and carcinoma in
situ one month prior to clinical studies. The ointment was
administered to the patients in conformity with the protocol of
Example 1 for 10 days. All patients were cured as confirmed by both
cytologist and cytopathologist.

Five patients were confirmed as Stage 0 cervical cancer. Three of the
patients selected had a familial history of cervical cancer. One
patient had a non-bleeding cervical ulcer 1.2.times.1.5 centimeters.
The diagnoses were made 15 days prior to clinical evaluations. The
ointment was administered to the patients in conformity with the
protocol of Example 1 for 10 days. All patients were cured, including
the one with the ulcer, said ulcer healing in 5 days. This was
confirmed by both cytologist and cytopathologist with no recurrences.

Five patients were selected who suffered from Stage 1 cervical cancer.
Three of the patients had bleeding ulcers. Four had a familial history
of cervical cancers. Diagnoses were made 12 days prior to the
commencement of clinical trials. The ointment was administered in
accordance with the protocol of Example 1 for 15 days. Four out of the
five patients were cured. The fifth did not respond. As for the
patients with the bleeding ulcers, the ulcers stopped bleeding on the
fourth day of the treatment. There were no recurrences in those who
were cured.

Five patients with Stage 1A cervical cancer were selected for therapy.
Four of the five had bleeding ulcers. Diagnoses were made 3 weeks
prior to clinical evaluations. Treatment entailed the protocol of
Example 1 over a 20 day period. Three of the patents were completely
cured. Those with bleeding ulcers, including the one who was not
cured, had all bleeding arrested on the sixth day of therapy. There
were no side effects in this group as with all the other groups cited
in the previous examples.

All of the 30 patients in these examples were in the age range of
40-60 years old.

Example 6

Treatment of Malignant Melanoma with Tetrasilver Tetroxide

A study group was formed of thirty-one patients between the ages of 48
and 78 years who had been diagnosed with malignant melanomas. The
diagnoses were confirmed by biopsy before the study was conducted. The
study group included four subgroups as discussed below.

Group I: Group I included fourteen female patients between the ages of
52 to 70 years. Biopsies confirmed superficial spreading melanomas
with a depth of invasion ranging from minor to 0.76 mm. The melanomas
were located in the interior extremities of the females. Each of the
patients began treatment with 200 mg of the 3% tetrasilver tetroxide
composition in lotion form applied twice a day. The fourteen patients
indicated that they had not previously been treated for the melanoma.

Group II: Group II included eight patients ranging between the ages of
65 to 78 years. A biopsy confirmed the presence of lentigo maligna
melanoma. Seven of the eight patients had a depth invasion of 0.76 mm
and one of them exhibited a 1.5mm depth of invasion. Each patient was
Caucasian and indicated that they had not previously received
treatment. Additional laboratory tests demonstrated no pain or
secondary effects in the patients. Each of the patients began
treatment with 200 mg of the lotion at 3% tetrasilver tetroxide
applied three times a day.

Group III: Group III included five patients ranging from ages 60 to 70
years old who had been diagnosed with nodular melanoma. The patients
had not previously received treatment. Three of the patients having a
depth invasion ranging from minor to 0.76 mm were placed in a subgroup
IIIA. Two patients exhibited a depth of invasion ranging 0.51 mm to
2.25 mm and were designated group IIIB. Group IIIB included patients
ranging in age from 64 to 68 years old. Two of these patients
exhibited discomfort associated with the lungs. Group III initiated
treatment with 200 mg of the lotion at 3% tetrasilver tetroxide
applied three times per day for 30 days.

Group IV: Group IV included four patients between the ages of 45 to 54
years old who had been diagnosed with acrolentiginous melanoma. Two of
the patients had received prior treatment with no success. The depth
invasion ranged between 1.51 to 2.25 mm. These two patients were
designated Group IVA. The remaining patients, Group IVB, exhibited a
depth of invasion ranging from minor to 0.76 mm. No discomfort was
reported by Group IV patients. Group IV began treatment with 200 mg of
the lotion at 3% tetrasilver tetroxide applied three times per day for
thirty days.

Results of the Melanoma Study

Each patient from Group I exhibited improvement by the sixth day of
treatment. The dark blue spots were losing color. By the eighth day of
treatment, the border on the injuries indicated improvement. By the
fourteenth day of treatment, none of the injuries exhibited
inflammation. On the fifteenth day of treatment, a biopsy was
conducted, which indicated that the malignant melanomas had been
replaced by normal epidermis while the dermis showed a decrease in the
malignant melanomas compared to the first biopsy. By day 22, the blue
spots had disappeared completely and the borders of the spots were not
visible at all. A new biopsy was conducted at the thirtieth day of
treatment, which showed the absence of the malignant melanomas from
both the dermis and epidermis.

The eight patients of Group II exhibited improvement by the eighth day
of treatment. The melanomas of the dermis had changed from black and/
or dark brown to a lighter color. By the fourth day, the injuries had
disappeared. At the fifth day, a new biopsy was conducted. The biopsy
indicated that the ratio of malignant melanomas to normal melanocytes,
i.e., benign melanomas, had decreased in relation to the pre-study
biopsy. The patients observed that the injuries had disappeared and
healed. A biopsy conducted at the twentieth day of treatment indicated
normal melanocytes. The patients have been monitored without any
changes from the above described results.

The patients of Group IIIA observed that their injuries had begun to
heal between the 9th and 10th day of the treatment. By the fourteenth
day of treatment, the grey colored spots were no longer visible. At
the sixteenth day of treatment, a new biopsy was obtained showing just
a few malignant melanomas and the appearing of normal melanocytes. By
the twenty-sixth day, the injuries were hardly visible, appearing only
as small scars. A biopsy obtained at the thirtieth day indicated the
presence of only normal melanocytes.

The patients of group IIIB exhibited few changes by the twenty-second
day of treatment. A biopsy obtained at the twenty-fifth day of
treatment indicated the presence of malignant melanomas. No
significant changes were reported at the thirtieth day. A biopsy
obtained at the this time indicated the presence of malignant
melanomas and minimum inflamation characteristics.

The patients of group IVA exhibited no change by the thirtieth day of
the treatment. A biopsy obtained at the this time indicated the
presence of malignant melanomas. The patients of group IVB, however,
exhibited changes to the border and color of the injuries by the
second day of treatment. By the sixth day of treatment, the injuries
were not visible and a biopsy acquired at the fifteenth day indicated
normal melanocytes.

Conclusions of the Melanoma Study

The present compositions, in ointment form, healed superficial
spreading melanomas within 8 to 14 days of treatment when the depth of
invasion was less than 0.76 mm. The biopsies indicated that the cancer
was eliminated. The compositions healed the lentigo-maligna having an
invasion depth between 0.76 mm to 1.5 mm. No discomfort or side
effects reported. The compositions also eliminated nodular melanoma
with a depth invasion of 0.76 mm or less within a sixteen day
treatment period. The compositions did not eliminate the nodular
melanoma having a metastasis condition with a depth of invasion of
1.51 mm to 2.25 mm within thirty days in this study.

The compositions of the invention were highly effective at treating
the acrolentiginous melanoma condition with a depth invasion ranging
from minor to 0.76 mm (without metastasis). The compositions
eliminated the cancer in 100% of these cases. The present compositions
were an effective treatment for the melanomas where chemotherapy and
surgical resources were ineffective. The compositions did not,
however, have a noticeable effect upon acrolentiginous melanoma with a
depth of invasion ranging from 1.5 mm to 2.25 mm within a period of 30
days of treatment.

Example 7

Preparation of a 3% Tetrasilver Tetroxide Ointment

A paraffinic hydrocarbon ointment was prepared by heating with
agitation a mixture comprising about 33 wt % heavy mineral oil and
about 67 wt % of petroleum jelly to a temperature of 70.degree. C. and
then dispersing in this mixture the finely divided tetrasilver
tetroxide powder sufficient to provide a 3% by weight concentration of
the oxide in the carrier. The mixture was then cooled to room
temperature with continuous stirring until the mixture was no longer
liquefied. It should be understood that the methods of this example
can be used in preparing the ointments and lotions of Examples 1-6, as
well as other compositions according to the invention, as will be
readily understood by those of ordinary skill in the art.

Example 8

Treatment of Melanoma

A male age 47 was diagnosed as having melanoma at least for three
years. He exhibited at least 15 blotchy brown lesions scattered on
both forearms. The size of the affected areas varied from 0.3 mm to
1.2 mm. The ointment of Example 7 was applied to the affected skin
areas of the arms at least once a day. After one week, the brown
blotches had been modified or replaced by pink ones. After 2 weeks,
the pink areas were gone. After 3 and 4 weeks, there was no evidence
of any melanoma.

Example 9

Treatment of Basal Cell Carcinoma

Two patients, one a male age 83 and the other a female age 63, were
treated in the same manner as in Example 8 for basal cell carcinoma.
The female had one large brown cancer growth varying from 0.6-1.4 mm
by 0.9-3.6 mm on top of the scalp. The male had several white head
pimples 0.3-0.7 mm. One was on the right ear. Five were on the scalp
and another was on the left leg. After one week the female observed
that her tumor had diminished to just a minor pink skin irritation.
After two weeks, the pink irritation had vanished. After four weeks
had elapsed, the skin was clear and normal in appearance. As for the
male, after one week all white head pimples had regressed to a pink
inflammation except for the ear, which appeared to be completely
healed. After the second week, all pink areas were gone, as well as
any indications of inflammation. After the third and fourth weeks, all
previously afflicted areas showed no evidence of basal cell carcinoma.

Example 10

Treatment of Squamous Cell Carcinoma

A male patient, age 74, was afflicted with squamous cell carcinoma.
The sores were on both arms, with five on the right and seven on the
left. Their appearance was oval shaped ranging from 2.5-5.5 mm in
length. This patient was treated in the same manner as in Example 8 by
applying the ointment of Example 7 to the affected skin areas. After
one week of treatment, all open sores had closed and were in the
process of healing. During the second week, sores continued to heal at
a rapid pace. Scabs were completely gone by the end of the second
week. At the end of the third week, only slight pink areas remained in
the place of the previous sores. By the end of the fourth week, there
were no traces of the previous affliction.

Example 11

Treatment of Basal Cell Carcinoma with Tetrasilver Tetroxide Ointment
3%

This study included twenty patients ranging in age from 45 to 65 years
old, all of whom had been diagnosed with basal cell carcinoma. The
diagnoses were confirmed by a pathologist via biopsy. All the patients
were Caucasian and commonly experienced a similar exposure level to
sunlight during their lives.

Group I: Group I included ten patients exhibiting injuries of less
then 1 cm in size with nodules that were bright and ulcerated. At the
time of diagnosis, no form of treatment had been implemented. The
patients had been evaluated for one month prior to treatment with
tetrasilver tetroxide composition of the invention formulated as an
ointment of 3% tetrasilver tetroxide. Each of the affected areas was
located in a region of the body exposed to sunlight. Each patient
began treatment with 200 mg tetrasilver tetroxide ointment 3%, three
times a day for thirty days.

Group II: Group II was formed by 10 patients having injuries larger
than 1 cm. The injuries were clinically present as nodules that were
ulcerated and with indurated edges. In each case, the period of
evaluation exceeded one month. Each patient began treatment with 200
mg tetrasilver tetroxide ointment 3%, three times a day for 30 days.

Results

Group I: Each patient within this group displayed a healing process of
the ulcerated nodules by the seventh day of treatment. It was no
longer possible to visualize the irritations and the bright color
became darker and more similar to the normal skin. By the twelfth day
of treatment, the ulcers were not visible at all and the scarring
process had begun. Controlled biopsies taken at the fifteenth day of
treatment indicated that the metaplastic cells from the injuries had
been replaced by normal appearing cells. The nodules were smaller than
0.5 cm. By the twenty-fourth day of treatment, the nodules were not
visible and only showed a light zone of papular tissue. The biopsy
control performed at 30 days revealed normal basal cellular tissue
with no signs of metaplastic cells. No recidivism was observed at the
thirty day examination.

Group II: Each patient in this group exhibited changes in the nodules
up to the fifteenth day of treatment when the beginning of the drying
process began and the nodules stopped exhibiting signs of peripheral
irritation areas. Biopsies performed on the 16th day of treatment
indicated a reduction in metaplastic cell with newly formed cells. The
ration of metaplastic cells to the normal cells began to invert by the
thirtieth day of treatment, and the ulcerated nodules had disappeared
and were only indicated by a few light zones. Biopsies performed on
the thirtieth day revealed that 80% of the affected tissue was
occupied by normal basal cells, but also indicated the presence of
metaplastic cells. The post-treatment evaluation at 30 days did not
clinically indicate any of the previous skin injuries.

In general, the results of the Group I and Group II studies indicate
that tetrasilver tetroxide ointment 3% was effective in the treatment
of basal cell carcinoma on injuries both smaller and larger than 1 cm.
The results, however, indicated that in the treatment of basal cell
carcinoma, it is most important to obtain and begin treatment as
rapidly as possible. Treatment was most effective the earlier the time
of diagnosis.

Example 12

Treatment of Dysplastic Nevi According to the Invention

Ten patients between the ages of 25 to 40 were clinically diagnosed by
biopsy with dysplastic nevi. The patients were divided into two
groups.

Group I had six patients with dermal injuries of 5 to 10 mm.

Group II had four patients with dermal injuries of more than 12 mm.
The illness was well developed on the skin.

A petroleum jelly containing 3 wt % tetrasilver tetroxide was applied
to both groups at a dosage of about 100 mg to all affected skin areas
of each patient twice daily. Observations of both groups were made for
a 30 day period to ensure there were no additional changes in the
condition.

Summary of Results

Group I: Within 36 hours of the onset of treatment, the color and size
of the injuries began to change, i.e., turn into smaller spots. By the
sixth day, the dysplastic nevi were no longer visible. By the eighth
day, a biopsy was conducted revealing new normal melanocytes. The
patients were evaluated for the duration of the period, with no
further changes reported.

Group II: By the fourth day of treatment, changes had started to
occur. The color from the spots had started to disappear, and they
were also turning smaller. By the fifteenth day, a new biopsy was
taken showing normal melanocytes, with the injuries no longer visible.
Patients did not experience any further changes in their condition
over the 30 days.

In conclusion, it is believed that early diagnosis resulted in better
and faster treatment results according to the invention, with no
significant adverse effects reported. Also, the larger the injuries,
the longer the treatment time required. The above test also showed
that the tetrasilver tetroxide treatment was effective to prevent the
malignant melanoma, since 90% of them derive from dysplastic nevi.

Example 13

Cytotoxicity Tumor Cell Proliferation Studies and Evaluations

An independent laboratory performed cytotoxicity tumor cell
proliferation studies and evaluations. A culture of leukemia K562 was
tested in vitro against media concentrations of tetrasilver tetroxide
at: 0.5, 1,5, 10, 50, 100, 500 and 1000 ppm.

The tetroxide was dispersed in pure dimethyl sulfoxide and then
diluted with Holipharm NPS buffer PH=7.4 (Holipharm International Co.,
Wilmington, Del.) in culture media to achieve the aforesaid final
assay concentrations. The acronym NPS refers to non phosphate non
saline. The culture media comprised RPMI 1640, 90% and fetal bovine
serum, 10%. The cancer cultures were obtained from a cell line
provided by the American Type Culture Collection (ATCC) and were
incubated at 37.degree. C. with 5% CO.sub.2 in air atmosphere. The
culture was a human chronic myelogenous leukemia, of cell line source
ATCCCCL-243.

As for the actual evaluation, cell proliferation analysis was based on
the ability of viable cells to cause alamar blue to change from
oxidized (non-fluorescent, blue) to reduced (fluorescent, red) form.
Details of the procedure are described in an article by S. Ansar Ahmed
et. al. in the Journal of Immunological Methods 170 (1994). The
results were as follows: IC.sub.50 (50% Inhibition Concentration)=2.2
ppm TGI (Total Growth Inhibition)=4.6 ppm LC.sub.50 (50% Lethal
Concentration)=6.0 ppm

The above methodology was applied to a human malignant melanoma cell
line of source ATCCHTB-70 having the cell name SK-MEL-5. The culture
media comprised 90% Minimum Essential Medium and fetal bovine serum,
10%. The results were as follows: IC.sub.50 (50% Inhibition
Concentration)=3.7 ppm TGI (Total Growth Inhibition)=4.9 ppm LC.sub.50
(50% Lethal Concentration)=6.5 ppm

The .sup.a IC.sub.50 (50% Inhibition Concentration) is the test
compound concentration where the increase from times in the number or
mass of treated cells was only 50% as much as the corresponding
increase in the vehicle-control at the end of experiment. The bTGI
(Total Growth Inhibition) is the test compound concentration where the
number or mass of treated cells at the end of experiment was equal to
that at time. The CLC (50% Lethal Concentration) is the test compound
concentration where the number or mass of treated cells at the end of
experiment was half that at time. Table 1.1 shows other results from
this study.

TABLE 1.1 Estimated IC.sub.50, TGI and LC.sub.50 Assay Name .sup.a
IC.sub.50 .sup.b TGI .sup.c LC.sub.50 Theor, Breast, T47D 1.1 ppm 1.6
ppm 2.6 ppm Tumor, Breast, MCF-7 3.0 ppm 3.9 ppm 5.0 ppm Tumor. Colon.
DLD-i 2.5 ppm 3.0 ppm 3.6 ppm Tumor, Prostate PC-3 7.7 ppm 14 ppm 25
ppm Tumor, Kidney, A498 1.8 ppm 2.4 ppm 3.1 ppm Tumor Pancreas MIA 2.0
ppm 4.0 ppm 8.1 ppm PaCa2 Tumor, Leukemia, 2.1 ppm 3.1 ppm 4.6 ppm
HL-60 Tumor Pancreas 4.8 ppm 6.4 ppm 8.6 ppm PANC-i Tumor. Liver.
HeDG2 2.6 ppm 3.5 ppm 4.7 ppm Tumor, Stomach 3.2 ppm 3.6 ppm 4.1 ppm
KATO III Tumor, Lung A549 4.8 ppm 5.8 ppm 7.0 ppm Tumor, Lung, pc-6
0.9 ppm 1.0 ppm 1.1 ppm Tumor, Lymphoma, 0.79 ppm 1.3 ppm 3.8 ppm
H33HJ-.JA1 Tumor, Lymphoma, 1.3 ppm 2.2 ppm 3.6 ppm U937

Although preferred embodiments of the invention have been illustrated
in the foregoing Summary, Detailed Description, and Examples, it will
be understood that the invention is not limited to the embodiments
disclosed, but is capable of numerous rearrangements and modifications
of parts and elements without departing from the spirit of the
invention. It will be further understood that the chemical and
pharmaceutical details of the compositions and methods of prevention,
treatment, or management herein may be slightly different or modified
by one of ordinary skill in the art without departing from the claimed
invention.



United States Patent 6,436,420
Antelman August 20, 2002
High performance silver (I,III) oxide antimicrobial textile articles

Abstract

Fibrous textile articles possessing enhanced antimicrobial properties
are prepared by the deposition or interstitial precipitation
of tetrasilver tetroxide (Ag.sub.4 O.sub.4) crystals within the
interstices of fibers, yarns and/or fabrics forming such articles.
Inventors: Antelman; Marvin S. (Rehovot, IL)
Assignee: Marantech Holding, LLC (East Providence, RI)
Appl. No.: 09/477,883
Filed: January 5, 2000

Current U.S. Class: 424/404 ; 210/758; 210/764; 424/402; 424/443;
424/449; 424/618; 514/495
Current International Class: A01N 25/34 (20060101); A01N 025/34 ()
Field of Search: 210/758,764 424/404,618,402,443,449 514/495
References Cited [Referenced By]
U.S. Patent Documents

2791518 May 1957 Stokes et al.
4101719 July 1978 Uetani et al.
4410593 October 1983 Tomibe et al.
5211855 May 1993 Antelman
5271952 December 1993 Liang et al.
5336499 August 1994 Antelman
5458906 October 1995 Liang
5676977 October 1997 Antelman
Primary Examiner: Page; Thurman K.
Assistant Examiner: Di Nola-Baron; Liliana
Attorney, Agent or Firm: Pennie & Edmonds, LLP
Claims


What is claimed is:

1. A fibrous textile article containing an antimicrobial agent
selected from the group consisting of tetrasilver tetroxide and
derivatives thereof interstitially deposited within said article, said
agent present in said article in an amount sufficient to impart
antimicrobial properties to said article.

2. The article of claim 1 wherein said agent is tetrasilver tetroxide.

3. The article claim 2 wherein said crystals are interstitially
deposited by interstitial precipitation.

4. The article of claim 3 wherein said textile article is a woven or
non-woven fabric.

5. The article of claim 4 wherein said agent is present within said
fabric at a level in the range of about 0.5 to about 50,000 weight
PPM, based on the weight of silver.

6. The article of claim 5 containing about 30 to about 10,000 PPM of
said agent.

7. The article of claim 3 wherein said antimicrobial properties are
sufficient to yield microbial inhibition zones extending beyond 1 mm
of fabric swatch borders as measured by AOAC test 972.04.

8. The article of claim 4 wherein said fabric is capable of
withstanding at least 100 hours of laundering without significant loss
of antimicrobial efficacy.

9. The article of claim 4 wherein said fabric is capable of
withstanding at least 600 hours of exposure to ultraviolet light
without significant loss of antimicrobial efficacy.

10. A method for producing a textile article having antimicrobial
properties, comprising: contacting the article with a first solution,
the first solution comprising silver; and contacting the article with
a second solution, the second solution comprising an amount of
oxidizing agent sufficient to deposit an antimicrobially active amount
of tetrasilver tetroxide within the article.

11. The method of claim 10 wherein said silver comprises silver
nitrate.

12. The method of claim 10 wherein said second solution comprises an
amount of alkali sufficient to provide a pH of at least about 13.

13. The method of claim 10 wherein said oxidizing agent is potassium
persulfate.

14. The method of claim 10 wherein said bath is heated to a
temperature in excess of 85.degree. C.

15. The method of claim 10 wherein said article contains from about
0.5 to about 50,000 weight PPM of precipitate, based on the weight of
silver.

16. The method of claim 15 wherein said article contains about 30 to
about 10,000 PPM of said precipitate.

17. A method for imparting antimicrobial properties to a fibrous
article, comprising: providing an aqueous dispersion of Ag.sub.4 O.sub.
4 crystals; and contacting the article with the aqueous dispersion.

18. The method of claims 17, wherein the article comprises low density
or loosely associated fibers.

19. The method of claim 18, wherein the article comprises a bandage,
gauze pad, or a laminated product.
Description


BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention relates to textile articles possessing antimicrobial
properties and a method for their preparation.

2. Description of Related Art

Textile articles which have been treated to render such articles
microbicidal to microorganisms coming in contact with the article are
known in the prior art. Such articles include those made from paper,
fibers, woven and non-woven textiles and like fabrics which are
designed for use in environments such as hospitals, food processing
plants, laboratories and other areas where maintenance of germ-free
conditions is essential.

For example, U.S. Pat. No. 2,791,518 discloses a method of imparting
microbicidal properties to articles such as textiles by immersing the
article in a first aqueous solution containing a water-soluble basic
nitrogen compound (ammonia) and a monovalent silver salt soluble in
said solution, followed by a second immersion in a second solution
containing a second salt capable of ion exchange with the silver salt
such that a monovalent silver salt precipitate is formed within the
article. The formed silver precipitate is sparingly water soluble and
imparts microbicidal properties to the articles so treated.

Similarly, U.S. Pat. No. 5,271,952 discloses a method of treating
fibers to render them electrically conductive as well as anti-
bacterial comprising immersing the fibers in a bath comprising an
aqueous solution of a source of divalent copper ions, a reducing
agent, sodium thiosulfate and a source of iodide ions, whereby copper
iodide is adsorbed into the fibers. Similar techniques for rendering
fibers conductive or resistant to bacteria involving the use of copper
compounds are disclosed in U.S. Pat. Nos. 4,410,593 and 5,458,906.

It has also been disclosed that materials such as chlorinated
hydantions may be grafted to textiles for the purpose of imparting
antimicrobial properties, ie Williams et al, 218.sup.th ACS National
Meeting (1999) Abstracts, Cell 32; C&EN September 6, page 36. However,
textiles so treated tend to suffer severe diminishment of
antimicrobial properties after as few as 5 hours of laundering and are
UV unstable over long durations of exposure.

SUMMARY OF THE INVENTION

The invention provides a fibrous textile article containing an
antimicrobial agent selected from the group consisting of tetrasilver
tetroxide and derivatives thereof interstitially deposited within said
article, said agent present in said article in an amount sufficient to
impart antimicrobial properties to said article.

The invention also provides a process for imparting antimocrobial
properties to a fibrous textile article comprising: a. providing an
aqueous solution containing a water soluble silver salt; b. contacting
said article with said solution for a period of time sufficient to
uniformly wet said article with said solution; c. immersing said
wetted article in a bath containing a second aqueous solution
containing a strong alkali and a water soluble oxidizing agent and
heating said bath for a period of time sufficient to interstitially
precipitate tetrasilver tetroxide within said article; and d. removing
said article from said bath.

Textile articles prepared in accordance with this invention,
particularly woven and non-woven hydrophilic fabrics, exhibit
outstanding antimicrobial resistance with respect to pathogens such as
bacteria, viruses, yeast and algae, are resistant to degradation upon
exposure to sunlight (ultraviolet light) and maintain their excellent
antimicrobial properties even after a number of launderings.

DETAILED DESCRIPTION OF THE INVENTION

Imparting antimicrobial properties to fiber and its derived textile
products is achieved in the instant invention by interstitial
deposition of the molecular crystal compound tetrasilver tetroxide,
i.e., silver (I, III) oxide. Said silver moiety is the subject of
several patents. U.S. Pat. No. 5,336,499, the disclosure of which is
incorporated herein by reference, describes the anti-pathogenic
properties of said silver oxide of formula Ag.sub.4 O.sub.4 and also
the mechanism of operation of the molecular device, based on a unique
crystal having two monovalent silver (Ag I) ions and two trivalent
silver (Ag III) ions in the molecule. The mechanism of killing
pathogens described in the patent is based on the differential silver
electronic activity between Ag (I) and Ag (III) resulting in
electrocution of pathogens, followed by binding chelation of
pathogenic surfaces.

U.S. Pat. No. 5,211,855 also discloses the use of Ag.sub.4 O.sub.4
crystals to kill pathogens in utilitarian water bodies such as
swimming pools.

An antimicrobial spectrum of Ag.sub.4 O.sub.4 is to be found in a
published article written by the instant inventor in the annual R&D
issue of Soap Cosmetics Chemical Specialties 1994, 70, 3 p. 52-59
entitled "Silver (II, III) Disinfectants", shown in Table 1. The
spectrum is based on specifications of the Association of Official
Analytical Chemists (AOAC).

TABLE 1 Antimicrobial Spectrum of Ag.sub.4 O.sub.4 MICROORGANISM MIC*
(PPM) Gram Negatives Escherichia coll 10231 2.50 Escherichia coll
25254 2.50 Enterobacter cloacae 13047 2.50 Pseudomonas aeruginosa 9027
1.25-2.50 Gram Positives Bacillus subtilis 6633 5.00 Micrococcus
lutena 9341 1.25-2.50 Staphylococcus aureus 0927 2.50-5.00
Staphylococcus aureus 27543 5.00 Staphylococcus epidermidis 12228
0.625 Streptococcus agalactiae 27956 1.25-5.00 Streptococcus faecium
10541 5.00 Streptococcus-pyogenes 7958 2.50 Yeast and Mold Candida
albicans 16404 2.50-5.0 Saccharomyces cerevisiae 2601 1.25 *MIC =
Miniinal Inhibitory Concentration.

In said article, reference is made to the fact that monovalent silver
is more anti-pathogenic than mercury which is more anti-pathogenic
than copper, based on their oligodynamic activity as articulated by J.
G. Horsfal in his "Principles of Fungicidal Action" (Chronica Botanica
1956). The relative efficacy of metallic moieties against pathogens
has been called the Horsfal series, and is as follows:

The present inventor has found that with respect to a Horsfal series
dedicated to silver, the relative efficacy against pathogens is as
follows:

The term "fibrous textile article" as used herein is intended to
encompass a wide variety of materials including paper, natural or
synthetic fibers, threads and yarns made from materials such as
cotton, rayon, wool, jute, nylon, polyesters, polyacetates,
polyacrylics as well as cellulosics in general. More particularly, the
term refers to fibers woven into a fabric such as knitting, and non-
woven hydrophilic fabrics or webbing used in anti-pathogenic
applications such as in the medical field, hospitals, biotechnology
and food and dairy processing. Exemplary textile products of this
genre include bandages, gauze, bandage pads, skin patches, work
clothes (both disposable and reusable), bed sheets, masks, dust
cloths, safety belts, surgical gowns, ambulance blankets, stretchers,
filter materials, diapers, underwear, pajamas, video display terminal
screens and the like.

For some antimicrobial applications, Ag.sub.4 O.sub.4 crystals may be
deposited within the interstices of fibrous articles by simply soaking
the article in an aqueous dispersion of the crystals or by combining
the crystals with a carrier medium and applying this composition to
the fibrous article. This method of physical incorporation of the
crystals is useful where the article is composed of low density or
loosely associated fibers such as bandage pads, gauze pads and loosely
non-woven products, and particularly laminated products wherein the
treated fibrous article is subsequently sandwiched between one or two
peelable layers which tend to keep the crystals trapped in the fibrous
article until ready for use. Also, antimicrobial paper products may be
made by simply mixing an aqueous dispersion of the Ag.sub.4 O.sub.4
crystals with paper pulp prior to calendaring the pulp.

However, physical incorporation of the crystals is less effective
where the treated article is a fiber or yarn or a higher density woven
or non-woven fabric, since the pre-formed crystals can not
sufficiently penetrate into the interstices of such articles. In such
cases, deposition of Ag.sub.4 O.sub.4 material via interstitial
precipitation is preferred.

Interstitial precipitation of Ag.sub.4 O.sub.4 material is
accomplished by first providing an aqueous solution of a monovalent
water soluble silver salt such as the nitrate perchlorate, acetate,
methanesulfonate or fluoride, most preferably silver nitrate. Next the
article to be treated, e.g., a fiber, yarn of a woven or non-woven
fabric, is thoroughly wetted with this solution such that the article
absorbs solution on fiber surfaces as well at one or more of the
interstices between fibrils forming the fiber, between fibers forming
the yarn or non-woven fabric, or between the weft and woof yarns
present in woven fabrics. Wetting may be accomplished by uniformly
spraying the article or more preferably by dipping the article in a
bath of the silver salt solution for a period of time sufficient for
the article to absorb the requisite amount of silver salt solution.

Next the wetted article is optionally squeezed to remove excess
solution and immersed in a heated bath containing a second aqueous
solution comprising a strong alkali and a water soluble oxidizing
agent, and heated for a period of time sufficient to cause reaction
leading to the interstitial precipitation of tetrasilver tetroxide
(Ag.sub.4 O.sub.4) crystal material in the interstices of the fibrous
article. Suitable alkalis for this purpose include sodium or potassium
hydroxide, with sodium hydroxide most preferred. Suitable oxidizing
agents include alkali metal persulfates, permanganates or
hypochlorites, but sodium and more preferably potassium persulfate is
the preferred oxidizer. Reaction in the bath is accomplished by
heating at a temperature of at least 85.degree. C., more preferably at
least 90.degree. C. for a period of time sufficient to maximize yield
of Ag.sub.4 O.sub.4, generally from about 30 seconds to about 5
minutes. After the reaction is completed, the treated article is
removed from the bath and may be washed several times with water to
remove soluble inpurities or unreacted reagant.

The quantity of Ag.sub.4 O.sub.4 material present in the resulting
article will generally be a function of the quantity of silver salt
sorbed by the article, which can vary depending on the nature of the
article, e.g., loose vs. tight weave fabrics or whether the fiber is
natural or synthetic, the former being more absorbtive of the silver
salt solution.

In general, the quantity of alkali present in the second bath should
be sufficient to maintain a strongly basic pH, i.e., about 13+, and
providing a slight molar excess of silver salt over oxidizing agent is
suitable to complete the reaction. Thus the content of tetrasilver
tetroxide interstitially precipitated within any given fibrous article
may be controlled by varying the concentration of the silver salt in
the solution used to first wet the article and appropriately adjusting
the quantities of alkali and oxidizing agent present in the immersion
solution at approximately stoichiometric levels.

The term "derivatives of Ag.sub.4 O.sub.4 " is intended to include
Ag.sub.4 O.sub.4 reaction products prepared by reacting Ag.sub.4 O.sub.
4 with suitable water soluble acids to give the corresponding Ag (II)
salts, e.g., reactions with fluoroboric acid or phosphoric acid to
give the Ag (II) fluoroborate or phosphate, as disclosed in U.S. Pat.
No. 5,107,295. Also included are divalent silver nitrate and divalent
silver halides prepared by reacting Ag.sub.4 O.sub.4 with nitric acid
or the corresponding haloacids, e.g. HBr, HI or HCl as disclosed in
U.S. Pat. No. 5,078,902. Trivalent silver derivatives such as Ag (III)
biguanide prepared in accordance with U.S. Pat. No. 5,223,149 are also
included.

Textile articles containing such derivatives would be prepared by
further contacting the Ag.sub.4 O.sub.4 containing article in an
additional step with an aqueous solution containing up to
stoichiometric amounts of the appropriate reagant(s) sufficient to
convert at least a portion of the Ag.sub.4 O.sub.4 to the Ag (II) or
Ag (III) derivative.

Textile articles containing such derivatives are less preferred for
the purposes of this invention because some derivatives may be
generally more water soluble than Ag.sub.4 O.sub.4, require a further
processing step in their manufacture and are less effective as
antimicrobial agents than Ag.sub.4 O.sub.4 as shown in the silver
Horsfal series described above. However, the Ag(II) or Ag(III)
derivatives of Ag.sub.4 O.sub.4 are useful as antimicrobial agents in
fabrics which are designed for a single use such as bandages or
disposable garments.

The content of the Ag.sub.4 O.sub.4 or its derivatives (based on
weight PPM silver) in the fabric may preferably range from as little
as 0.5 weight PPM up to about 50,000 weight PPM, based on the weight
of the textile article. The minimum content should be sufficient to
kill pathogens from which protection is sought, whereas the maximum
content is dictated by factors such as economy and affect on fabric
properties. Generally speaking, the higher the silver content, the
more effective will be the antimicrobial properties of the fabric. For
most applications, silver content in the range of from about 30 to
about 10,000 weight PPM will provide satisfactory antimicrobial
properties.

Antimicrobial properties are evaluated in accordance with this
invention using the Association of Official Analytical Chemists (AOAC)
test method 972.04, which is used primarily to evaluate the
bacteriostatic activity of laundry additive disinfectants. In this
test, a square or rectangular sterile swatch of fabric is pressed into
a petri dish containing a layer of nutrient agar which has been
inoculated with a pathogen. Following a fixed period of incubation,
each fabric sample is evaluated by measuring the clear zones adjacent
the four sides of each test swatch as an index of antimicrobial
activity. The presence of clear zones along all four sides of the
swatch is indicative of antimicrobial activity, rated 4/4. The width
of the clear zones in millimeters is reasonably indicative of the
degree of antimicrobial activity.

The following examples are illustrative of the invention.

EXAMPLE 1

A swatch of virgin nylon webbing was immersed in an aqueous solution
containing dissolved silver nitrate at a concentration of about 100
PPM silver maintained at room temperature. After 30 seconds immersion
time, the swatch was removed from this solution and immersed in a hot
aqueous solution containing 7.2 g/liter each of NaOH and sodium
persulfate, which solution is then boiled for one minute
(95-100.degree. C.). The swatch was then removed from the boiling
solution, washed with water and dried. There was perceived a hardly
visible tan coating on the swatch fibers. The content of Ag.sub.4
O.sub.4 in the fabric swatch, measured as silver, was 89 weight PPM as
verified by gravimetric analysis.

EXAMPLES 2-6

Example 1 was repeated except that the concentration of silver nitrate
in the first solution and reagants in the second solution were varied
to provide the following Ag content in the fabric swatch:

SOLUTION (PPM Ag) FABRIC (PPM Ag) Ex 2 890 35* Ex 3 890 541 Ex 4
10,000 3970 Ex 5 10,000 9140 Ex 6 10,000 9670 *Nylon fabric for this
test was of a tighter weave than that used in Example 1 which accounts
for the lower silver take up.

AOAC antipathogenic tests on these textiles were performed by an
independent laboratory which was licensed by a State environmental
regulatory body. The marker organisms used in conformity with AOAC
test method 972.04 were Pseudemonas Aeroginosa (PA) as the Gram
negative bacteria marker, and Staphylococcus Aureus (SA) for Gram
positive bacteria.

The tests were conducted in terms of inhibition of cultures of the
bacteria. Two swatches were used for the tests in contact with the
cultures. The swatches were 1.5 inches wide and weighed about 69 mg./
cm.sup.2. Each swatch had four sides, and two swatches were used with
each representative culture so that a total of eight trials were
reflected with each bacterium. An 8/8 inhibition would indicate 100%
efficacy. However, the test protocol went beyond the specifications of
the AOAC method insofar that the actual average inhibition zone width
in millimeters was recorded for both swatches tested. These results
were then combined with the aforementioned anti-microbial spectrum
shown in Table 1 which includes the marker bacteria of the AOAC tests,
and extrapolated. The conclusion was that the preferred embodiments of
the invention were 100% effective against all of the microbes shown in
Table 1 and against salmonella and the AIDS virus based on the
previous independent results obtained with silver (I, III) oxide.

Representative results with nylon fabric are shown in Table 2.
Generally, the degree of microbial activity varies directly with the
silver concentration. In addition to the high performance anti
microbial properties of the fabrics, they withstood wear and could be
considered permanent insofar that tested fabrics withstood 100 hours
of laundering and 600 hours of ultra violet exposure. Laundering is
evaluated using hot water and detergent using the standard test of the
American Association of Textile Chemists (AATC).

TABLE 2 Antimicrobial Performance of Precipitated Ag.sub.4 O.sub.4
Inhibition Inhibition Example Silver Zone-SA Zone-PA Inhibition Number
PPM (mm) (mm) Index 1 89 3.2 1.3 8/8 2 35 1.8 2.0 8/8 3 541 5.5 5.1
8/8 4 3970 5.8 2.6 8/8 5 9140 5.8 2.8 8/8 6 9670 6.1 4.8 8/8

EXAMPLE 7

An independent medical researcher in Israel obtained a very virulent
strain of Staph from a patient at the Shaarei Tzedek Hospital in
Jerusalem. The patient subsequently died from infection. This strain
was evaluated as more virulent than any of the other Staph
microorganisms listed in Table 1 by the pathology staff at the
hospital. This Staph strain was utilized as the Staph source, and the
otherwise exact test protocol described above was repeated. The silver
concentration of the test swatch was found to be 9,138 PPM. Only one
test swatch was used for the Staph evaluation. It tested at 4/4 with a
much diminished average inhibition zone of 0.50 mm, which was to be
expected for the more virulent strain. The values were extrapolated
for all Gram positive bacteria listed in Table 1. It was concluded
that precipitated Ag.sub.4 O.sub.4 was capable of inhibiting all of
the listed Gram positive bacteria. The extrapolation took into
consideration a theoretical calculation of the reduction of the Staph
inhibition zone, were the conventional Staph aureus organisms to
display the listed MIC range of 2.5-5.0 PPM. Since the inhibition zone
is inversely proportional to the MIC, one can calculate that the MIC
for the virulent Staph strain was 40.5-61.0 PPM. By applying the same
reasoning to the Gram Negative microorganisms for their PA marker, one
can claim inhibition as well for all Gram Negative bacteria listed in
Table 1 by Ag.sub.4 O.sub.4.

EXAMPLE 8

The method of Example 1 was repeated with larger amounts of webbing
utilizing one-foot lengths. Accordingly, webbing was obtained having
respectively 4730 and 9430 PPM of silver. These materials were dyed
orange, and the dye completely covered and hid the brown/black color
imparted to the virgin webbing by the tetroxide at these relatively
high levels. After dyeing, swatches of the webbing were cut from the
master rolls and were then evaluated in the same manner as described
above by exposure to Staph aureus. All swatches indicated an 8/8
score, with average inhibition zones of 6.3 and 6.0 respectively for
the 4730 and 9430 PPM samples. Lengths of the dyed webbing were
subjected to 100 hours of laundering in accordance with the AATC
method, after which bacteriostatic efficacy was again evaluated.
Visual inspection after laundering revealed frayed webbing.
Nevertheless, both materials exhibited an 8/8 score with an
improvement in inhibition zones to 7.0 for Staph aureus. This
indicated that the laundry wear tended to expose fresh surface of
tetroxide from the fabric interstices. Swatches were again taken from
these materials and exposed to 600 hours of ultra violet light in a
weathering test. Evaluations of the UV exposed samples with Staph
aureus again indicated scores of 8/8 for both concentrations of
silver, with inhibition zones of 6.5 and 4.6, respectively for the
4730 and 9430 PPM silver concentration webbing. Accordingly, ultra
violet exposure did not interfere with bacteriostatic activity.

Comparative Example

Monovalent silver iodide was interstitially precipitated within nylon
fabric such that the fabric contained 4895 PPM silver. Test swatches
were prepared and evaluated against SA and PA pathogens by the test
procedure described above. The Inhibition Index for SA was 7/8 and for
PA was 0/8. In addition, the SA inhibition zone was only about 0.5 mm.

Fabrics treated in accordance with this invention hold promise for
many antimicrobial applications ranging from preventing jock itch when
applied to athletic supporters to preventing scabies and bed sores
with treated bed sheets or hospital gowns used in nursing homes and
hospitals.



United States Patent 6,258,385
Antelman July 10, 2001
Tetrasilver tetroxide treatment for skin conditions

Abstract

The invention relates to the use of electron active molecular crystals
comprising tetrasilver tetroxide (Ag.sub.4 O.sub.4) for the treatment
and cure of dermatological skin conditions (diseases) ranging from
dermatitis, acne and psoiasis to herpes and skin ulcers.
Inventors: Antelman; Marvin S. (Pehovot, IL)
Assignee: Marantech Holding, LLC (Providence, RI)
Appl. No.: 09/552,172
Filed: April 18, 2000
Related U.S. Patent Documents

Application Number Filing Date Patent Number Issue Date
296998 Apr., 1999

Current U.S. Class: 424/618 ; 424/405; 424/439; 514/859; 514/861;
514/863; 514/925; 514/928; 514/931; 514/934; 514/937; 514/938; 514/969
Current International Class: A61K 47/02 (20060101); A61K 8/02
(20060101); A61K 8/19 (20060101); A61Q 1/12 (20060101); A61K 33/24
(20060101); A61K 33/26 (20060101); A61K 33/32 (20060101); A61K 33/34
(20060101); A61K 33/38 (20060101); A61K 9/06 (20060101); A61K 033/38
(); A61K 009/00 ()
Field of Search: 424/405,489,618
514/859,861,863,925,928,931,934,937,938,969
References Cited [Referenced By]
U.S. Patent Documents

4828832 May 1989 De Cuellar et al.
5017295 May 1991 Antelman
5073382 December 1991 Antelman
5078902 January 1992 Antelman
5089275 February 1992 Antelman
5098582 March 1992 Antelman
5211855 May 1993 Antelman
5223149 June 1993 Antelman
5334588 August 1994 Fox, Jr.
5336416 August 1994 Antelman
5336499 August 1994 Antelman
5571520 November 1996 Antelman
5676977 October 1997 Antelman
5772896 June 1998 Denkewicz, Jr. et al.

Other References

The Merck Manual (16th Ed. 1992), `Dermatologic Disorders`, pp.
2399-2460.* .
Dorland's Illustrated Medical Dictionary (28th Ed. 1994), pp.
351,759-60.* .
Remington's Pharmaceutical Sciences (17th Ed. 1985), pp. 1573-1575,
1585-1602.* .
Antelman, Marvin S.; "Silver (II,III) Disinfectants"; Soap/Cosmetics/
Chemical Specialties, Mar. 1994, pp. 52-59. .
Antelman, Marvin S.; Abstracts of the American Chemical Society;
1992(203). .
Antelman, Marvin S.; "Anti-Pathogenic Multivalent Silver Molecular
Semiconductors"; Precious Metals; 1992(16); pp. 141-149. .
Antelman, Marvin S.; "Multivalent Silver Bactericides"; Precious
Metals; 1992(16); pp. 151-163..

Primary Examiner: Dees; Jose' G.
Assistant Examiner: Choi; Frank
Attorney, Agent or Firm: Pennie & Edmonds LLP
Parent Case Text


This Application is a Continuation-In-Part of application Ser. No.
09/296,998, filed Apr. 22, 1999, now abandoned.
Claims


What is claimed is:

1. A method for treating dermatological skin disease comprising
applying a composition comprising a therapeutically effective amount
of tetrasilver tetoxide directly to the affected skin of a patient in
need of treatment, said composition being free of added oxidizing
agent, wherein the dermatological skin disease is selected from the
group consisting of eczema, psoriasis, dermatitis, ulcers, shingles,
rashes, bedsores, cold sores, blisters, boils, herpes, acne, pimples,
warts, and a combination thereof.

2. The method of claim 1, wherein the tetrasilver tetoxide is
dispersed in a carrier medium at a concentration of from about 50 to
250,000 wt PPM, based on the weight of said carrier medium.

3. The method of claim 2 wherein said concentration is from about 400
to 50,000 PPM.

4. The method of claim 2 wherein said carrier medium comprises
petroleum jelly.

5. The method of claim 1, wherein said skin disease includes cold
sores.

6. The method of claim 1, wherein said skin disease includes herpes.

7. The method of claim 1, wherein said skin disease includes shingles.

8. The method of claim 1, wherein said skin disease includes acne.

9. The method of claim 1, wherein said skin disease includes
psoriasis.

10. The method of claim 1, wherein said skin disease includes
dermatitis.

11. The method of claim 1, wherein said skin disease includes disease-
induced skin ulcers.

12. The method of claim 1, wherein said skin disease includes eczema.

13. The method of claim 1, wherein said tetrasilver tetroxide is
applied in powered form.

14. The method of claim 1, wherein said composition is applied to the
skin at a dosage level of from about 10 to 500 mg per cm.sup.2 of skin
surface.

15. The method of claim 1, wherein said skin disease includes warts.

16. The method of claim 1, wherein said skin disease includes pimples.

17. The method of claim 1, wherein said skin disease includes
blisters.

18. The method of claim 1, wherein said skin disease includes
bedsores.

19. The method of claim 1, wherein said skin disease includes rashes.

20. The method of claim 1, wherein said composition is applied in non-
sprayable form.
Description


BACKGROUND OF THE INVENTION

The present invention relates to the use of electron active molecular
crystals comprising tetrasilver tetroxide (Ag.sub.4 O.sub.4) for the
treatment and cure of dermatological conditions or diseases.

The present invention is related to concepts previously elucidated and
disclosed in U.S. Pat. No. 5,336,499 (1994); U.S. Pat. No. 5,571,520
(1996) and U.S. Pat. No. 5,676,977 (1997). Said concepts have also
been published in an article entitled "Silver (II, III)
Disenfectants" (Soap/Cosmetics/Chemical Specialties, 1994; March, pp.
52-59).

The aforementioned references disclose that said tetrasilver tetroxide
devices kill and/or inhibit a wide variety of pathogens ranging from
gram positive and negative bacteria (e.g., E coli and Staphylococcus
aureus), algae and mold (e.g., Chorella and Candida albicans) and the
AIDS virus. Said references also contain detailed descriptions of the
mechanism via which said molecular crystal devices operate. The
instant inventor also presented his subsequent results and concepts at
a Seminar entitled "Incurable Diseases Update" (Weizmann Institute of
Science, Rehovot, Israel, Feb. 11, 1998). The title of his
presentation was "Beyond Antibiotics, Non Toxic Disinfectants and
Tetrasil Mademark of applicant for the tetroxide)".

In the aforecited article it was shown that the effects of the
electron transfer involved with respect to the tetroxide,
phenomenally, rendered it a more powerful germicide than other silver
entities. The instant inventor holds patents for multivalent silver
anti microbials, e.g., U.S. Pat. No. 5,017,295 for Ag(II) and U.S.
Pat. No. 5,223,149 for Ag (III); and while these entities are stronger
anti microbials than Ag (I) compounds they pale by comparison to the
tetroxide and so does colloidal silver which derives its germicidal
properties from trace silver (I) ions it generates in various
environments. Accordingly, the oligodynamic properties of these
entities may be summarized as follows, which is referred to as the
Horsfal series:

The other unique property of the tetroxide was that it did not stain
organic matter such as skin in like manner as Ag(I) compounds do. In
addition, it was light stable.

The main object of this invention is to utilize tetrasilver tetroxide
molecular devices to cure dermatological diseases or conditions.

Another object of this invention is to use tetrasilver tetroxide
molecular devices to control those dermatological conditions or
diseases which cannot be cured completely by said devices.

Still another object of this invention is to use tetrasilver tetroxide
molecular devices to reduce the time of affliction of dermatological
conditions or diseases.

Still another object of this invention is to utilize said devices in
the aforesaid dermatological applications without staining the skin.

SUMMARY OF THE INVENTION

This invention relates to a molecular scale device comprising a single
crystal of tetrasilver tetroxide (Ag.sub.4 O.sub.4). Several trillion
of these molecules may be employed in various pharmaceutical
formulations and therapies to effectuate the treatment and cure of
various dermatological conditions and diseases. These conditions and
diseases vary and include eczema, psoriasis, dermatitis, disease
induced skin ulcers, undefined tropical diseases, shingles, rashes,
bedsores, cold sores, blisters, boils, herpes simplex, acne, pimples,
skin chafing, cracking, itchiness, skin peeling, and warts.

More particularly, the invention relates to a method for treating
dermatological skin disease comprising applying a composition
comprising tetrasilver tetroxide directly to the affected skin areas,
said composition being free of added oxidizing agent.

Even more particularly, the invention relates to a method for treating
dermatological skin conditions selected from the group consisting of
eczema, psoriasis, dermatitis, ulcers, shingles, rashes, bedsores,
cold sores, blisters, boils, herpes, acne, pimples, skin chafing,
cracking, skin itch, skin peeling and warts comprising applying
tetrasilver tetroxide directly to the affected skin area.

DESCRIPTION OF THE INVENTION

The crystal lattice of the Ag.sub.4 O.sub.4 molecular device operates
against pathogens by transferring electrons from its two monovalent
silver ions to the two trivalent silver ions in the crystal
contributing to the death of pathogens by traversing their cell
membrane surface. This in effect "electrocutes" the pathogens. The
electrons are forced out of their balanced crystals by such labile
groups as NH, NH.sub.2, S--S and SH comprising pathogen cell membrane
surface. However, normal cells will not be affected because they do
not proliferate fast enough to expose these labile bonds.

The K.sub.A of Ag.sub.4 O.sub.4 is 7.9.times.10.sup.-13, therefore the
molecule will not be disturbed unless more stable complexes are formed
with such ligands as those comprising the pathogen cell membrane
surface in a dynamic state. Indeed the end result of the electron
transfer, which is a redox reaction, results in the monovalent Ag ions
being oxidized to Ag(II) and the trivalent Ag ions being reduced to
the same end product, Ag(II). Accordingly, the well-known affinity for
monovalent silver for certain elements such as sulfur and nitrogen is
far exceeded here, for divalent silver will not merely bind to these
elements as does silver, but will actually form chelate complexes with
their ligands. The molecular crystal attraction for the cell membrane
surfaces is thus driven by powerful covalent bonding forces.

The electron transfer can be depicted by the following redox half
reactions: ##EQU1##

It was found by rigorous testing that silver tetroxide was
comparatively non toxic. Since said oxide was effective at PPM
concentrations in killing pathogens, commercial concentrates were
formulated with 2% of the tetroxide. For acceptance of said oxide in
commerce for which an EPA registration number was obtained (no.
3432-64), it was necessary for the oxide to undergo a series of
toxicity tests, for which a 3% concentrate was used and evaluated by a
certified laboratory employing good laboratory practice (GLP)
according to the Code of Federal Regulations.

The results were as follows:

Acute Oral Toxicity LD.sub.50 Greater than 5,000 mg./Kg. Acute Dermal
Toxicity LD.sub.50 Greater than 2,000 mg./Kg. Primary Eye Irritation
Mildly irritating Primary Skin Irritation No irritation Skin
Sensitization Non Sensitizing

Subsequent evaluations showed that unless persons were prone to silver
allergies, the pure tetroxide could be applied to the skin without any
ill effects or evidence of irritation, despite the fact that said
oxide is a powerful oxidizing agent. This can perhaps be explained by
its stability manifested by its aforecited K.sub.A.

It was previously postulated in earlier patents relating to the
various uses of the oxide that it was required to use it in
combination with an excess of strong oxidizing agent such as a
persulfate in order to effectively kill pathogens. However, the
additional presence of oxidizing agent tends to be irritating to the
skin. It has been found in accordance with the present invention that
the additional oxide is unnecessary and in fact undesirable for the
purpose of treating the skin diseases described herein. Therefore, the
present invention contemplates application of the oxide to the skin in
the absence of the additional oxidizer and the use of formulations
which are free of added additional oxidizing agent such as a
persulfate.

A preferred mode of application of the oxide of the invention is as a
topical agent, either directly as a powder or in non-sprayable or
sprayable form. Non-sprayable forms can be semi-solid or solid forms
comprising a carrier indigenous to topical application and having a
dynamic viscosity preferably greater than that of water. Suitable
formulations include, but are not limited to, suspensions, emulsions,
creams, ointments, powders, liniments, salves and the like. If
desired, these may be sterilized or mixed with auxiliary agents, e.g.,
thixotropes, stabilizers, wetting agents, and the like. Preferred
vehicles for non-sprayable topical preparations include ointment
bases, e.g., polyethylene glycol-1000 (PEG-1000); conventional
ophthalmic vehicles; creams; and gels, as well as petroleum jelly and
the like. These topical preparations may also contain emollients,
perfumes and/or pigments to enhance their acceptability for various
usages.

Where the oxide is applied to the skin combined with a carrier such
one or more of the carriers described above, it may be combined with
the carrier at a level of from about 50 to 250,000 PPM, more
preferably from about 400 to 50,000 PPM, based on the weight of the
carrier, and applied to the skin 1 to 3 times per day until the
condition is cured or satisfactorily controlled. Generally, the
composition may be applied at a dosage level of from about 10 to 500
mg per cm.sup.2 of skin surface.

Accordingly, the oxide was used directly in powder form as well as in
several compounded formulations for treating a wide assortment of skin
conditions and diseases. Success was achieved in all cases except for
certain stubborn nail fungi. After much experimenting it was found
that the best carrier for a commercial product was white petroleum
jelly.

Other objects and features of the present invention shall become
apparent to those skilled in the art when the present invention is
considered in view of the accompanying examples. It should, of course,
be recognized that the accompanying examples illustrate preferred
embodiments of the present invention.

EXAMPLE 1

A female, age 28, resident of Central America, had a red rash caused
by an unidentified dermatological tropical disease on her thigh. The
condition was cured by a light dusting of Ag.sub.4 O.sub.4 crystals on
the area. Similar occurrences in the past to the subject failed to be
cured by other dermatological preparation sold as cures for said
condition.

EXAMPLE 2

A female, age 27, had a fungus infection in her navel. She was cured
by direct application of Ag.sub.4 O.sub.4 to the affected area within
24 hours.

EXAMPLE 3

A female in her early thirties had suffered from recurrent cold sores
for five years. The subject stated in a written communication, "I have
tried every over-the-counter medication for this ailment without even
marginal success. I even tried the five times a day for five days
herpes medication that my doctor prescribed with disappointing
results." Subject tried various concentration of Ag.sub.4 O.sub.4
dispersed in petroleum jelly. All formulation improved the severity
and duration of the herpes simplex. Subject was given a final
formulation of 10,000 PPM Ag.sub.4 O.sub.4 dispersed in white
petroleum jelly. In many instances quick application of the ointment
resulted in disappearance of the cold sore the next day. Otherwise if
not caught quickly the sore could be contained within 36 hours which
was a vast improvement over the previous treatments.

EXAMPLE 4

An 82-year-old female had suffered six months from an external vaginal
itch which defied treatment. Application of Ag.sub.4 O.sub.4 ointment
(as described in Example 3) cured the condition.

EXAMPLE 5

Twenty-two samples of Ag.sub.4 O.sub.4 ointment were distributed to
individuals who were suffering from the herpes simplex. They all
applied the ointment. While there was no attempt made to record the
exact condition of the herpes subjects, all 22 cases were cured within
48 hours.

EXAMPLE 6

Having achieved success against herpes simplex, it was decided to test
Ag.sub.4 O.sub.4 ointment against shingles which is caused by herpes
zoster. Accordingly, a 67-year-old male applied the ointment three
times a day for two days, after which time the shingles condition was
completely gone.

EXAMPLE 7

Two individuals, one male, the other female, ages 33 and 48, who were
suffering from external acne condition, treated their skin three times
a day with Ag.sub.4 O.sub.4 ointment. They were completely cured after
two days of applying the ointment.

EXAMPLE 8

Fifteen patients with an age ranging from 30 to 35 which were
diagnosed as having oral viral herpes were arranged in two groups.
Group I consisted of five patients which suffered from severe oral
viral outbreaks with a recurring frequency of 21-28 days. The sizes of
the herpes sores ranged from 3.5-5.0 mm. Group II consisted of ten
patients who suffered from normal oral viral outbreaks with a
recurring frequency of 28-42 days. The sizes of the herpes sores
ranged from 1.25-1.75 mm. Both groups applied tetrasilver tetroxide
ointment to the effected areas with 50-200 mg. of ointment containing
3% tetrasilver tetroxide. Group I applied the ointment (within 12
hrs.) after the herpes sores broke through the skin and blistered.
Group II was divided into two subgroups. Group IIA applied the
ointment (within 12 hrs.) after the herpes sores broke through the ski
and blistered. Group IIB applied the ointment 4-12 hrs. before the
herpes sores broke through the skin and blistered. Application was
twice daily. Patients reported daily on the pharmacological effects.
Sizes of the herpes growth were observed on a daily basis for five
days and frequency of reoccurrence was recorded.

Summary of Results

Group I: Over a period of 24-48 hours all of the patients observed the
herpes sore regress and dry out. By day three the sores were not
visible and the skin was healed. All patients exhibited a longer
recurrence time from 32-44 days excluding one patient who did not have
a recurrence for eight months. The sizes of the herpes upon recurrence
were significantly smaller at 2.2-3.5 mm.

Group IIA: Over a period of 24-48 hours all the patients observed the
herpes sore regress and dry out. By the end of day the sores were not
visible and the skin was healed. All patients exhibited a longer
recurrence time from 34-55 days. The sizes of the herpes upon
reoccurrence were significantly smaller at 0.8-1.4 mm.

Group IIB: Over a period of 12-24 hours all the patients observed that
the herpes sore was retained and never broke through the skin as a
blister. By the end of day two there were no signs of the herpes sore
at all. There was not even the slightest amount of discomfort around
the area where the blisters would have flourished. All patients
exhibited a longer recurrence time from 36-62 days. The sizes of the
herpes upon recurrence were 0.7-1.6 mm.

Conclusions

Tetrasilver tetroxide used as a topological ointment:

1. Eliminates oral viral herpes sores within a period of 48 hours from
the time of the first application.

2. Extends the recurrence period of the viral herpes breakout cycle.

3. Prevents the herpes virus from breaking through the skin when used
before an outbreak occurs.

EXAMPLE 9

Twenty eight patients in the age group ranging from 45 to 65
presenting with diabetes--induced foot ulcers were arranged in two
groups. All of the patients were taking insulin injections and were
diagnosed as Type I insulin dependent.

Group I consisted of fourteen patients where culture swabs of the
ulcerated skin indicated the presence of bacteria (infection). Group
II consisted of fourteen patients where culture swabs of the ulcerated
skin did not indicate the presence of abnormal amounts of bacteria,
(no infection).

The patients in each group were treated by applying 200 mg of a
petroleum jelly containing 3 wt % tetrasilver tetronide twice daily to
the ulcerated sores for a 30 day period. Daily evaluations of the skin
condition were conducted by a dermatologist.

Summary of Results

Group I: Within 48 hours of the onset of treatment the sores on the
feet of all patients began to dry out. After 72 hours, the ulcers on
all patients started to heal at the borders. By the fourth day,
inflammation of the diseased tissue eased and by the sixth day the
ulcers were completely dry with no surface secretions. By the tenth
day the ulcers on all patients had completely disappeared. Lab tests
indicated no sign of infection on the feet of any patient.

Group II: Within 24 hours of the onset of treatment the sores on the
feet of all patients began to dry out and heal at the borders with no
secretion. By the third day the sores on all patients were covered
with new healthy tissue. By the tenth day the ulcers had healed and
completed the process of forming scar tissue by 80%. At day 14 of the
treatment all of the ulcers were 100% healed with no sign of
infection.

Continuous monitoring of both groups over the 30 day period indicated
no reappearance of the ulcers.

The above testes demonstrate that tetrasilver tetroxide treatment is
effective in both curing infections associated with diabetes-induced
ulcers and in healing the ulcers themselves. The tetroxide accelerated
the neovascularization` process of the affected tissue.

EXAMPLE 10

Twenty patients ranging from age 8 months to 12 years were clinically
diagnosed as suffering from atopic dermatitis involving inflamed
lesions of the face and extremities, but without bacterial
involvement. These patients were previously treated by the application
of topical steroids to the affected skin areas, which was
discontinued. The patients were divided into two groups.

Group I consisted of ten randomly selected patients. A petroleum jelly
containing 3 wt % tetrasilver tetroxide was applied at a dosage of
about 100 mg. to all affected skin areas of each patient twice daily
for a period of days. Daily evaluation of the skin condition was made
by a dermatologist.

Group II consisted of a control group comprising the remaining ten
patients. This group was treated by twice daily application of about
100 mg of pure petroleum jelly which was free of added tetrasilver
tetroxide to the affected skin areas.

Summary of Results

Group I: Within 12 hours of the onset of treatment the lesions on all
patients began to show healing and drying and no longer exhibited
prurito in the affected skin areas. Within 24 hours of the onset of
treatment, signs of irritation of the skin areas had subsided and
after 24 hours signs of irritation had disappeared and the lesions
were no longer visible. Treatment on all patients was discontinued
after 5 days, but the group was assessed daily for any recurrence of
the lesions. Two of the patients presented a reappearance of lesions
by the twenty-fourth day, but these lesions were smaller and less
irritating than the original lesions. Treatment was resumed on these
two patients and after 24 hours the lesions had disappeared.

Group II: At 12 hours after the onset of the application of pure
petroleum jelly to the affected skin areas there were no signs of
improvement of the skin. Over the next 23 day period the lesions
gradually became more irritated with no sign of healing of the atopic
dermatitis.

The above tests demonstrate that the tetrasilver tetroxide treatment
is effective in most patients in healing atopic dermatitis within 24
hours of the commencement of treatment and appears to halt the self
immunological reaction of atopic dermatitis at the local level. It is
also effective in reversing disease when it recurs, increasing the
period of recession of the condition.

EXAMPLE 11

Twenty four patients between the ages of 13 and 40 years were
diagnosed as suffering from psoriasis, exhibiting irritation,
scaliness and both the Auspitz sign and the Koebner phenomenon. All
patients had been previously treated with topical steroids. The
patients were divided into two groups.

Group I consisted of 12 patients where psoriasis was diagnosed less
than 60 days prior to treatment A petroleum jelly containing 3 wt %
tetrasilver tetroxide was applied to affected skin areas twice a day
over a 30 day period and each patient was evaluated twice daily by a
dermatologist.

Group II consisted of 12 patients who were diagnosed more than 60 days
prior to treatment. Disease in this group was more severe than Group I
and most had been suffering from psoriasis for many years, exhibiting
extensive disease on their backs. This group was treated by the same
protocol as Group I and was evaluated three times daily by a
dermatologist.

Summary of the Results

Group I: By the tenth day of treatment the psoriatic plates and
inflamed areas of the treated skin started to heal. By the twentieth
day the Auspitz sign had disappeared on all patients. By day 27 the
psoriatic plates present in the diseased skin of all patients had
disappeared. By day 35 the skin on all patients appeared to be healed
and the pigmentation process of the skin had been initiated.

Group II: By the twentieth day of treatment, the healing process on
all patients had commenced as evidenced by the reduction of psoriatic
plates and appearance of new tissue. By day 28 the diseased areas were
of smaller sizes and the Auspitz sign was no longer visible. By day
30, the Koebner phenomenon had disappeared on all patients. By day 35,
the psoriatic plates were barely visible on all patients.

Continued observation of both groups showed no further changes in
their condition after treatment was halted.

The above tests demonstrate that topical application of tetrasilver
tetroxide to the affected skin areas of psoriasis sufferers
effectively heals and/or controls this disease.

EXAMPLE 12

Twelve Caucasian patients between the ages of 45 to 65 presented with
infected bleeding skin ulcers associated with diagnosed
Rabdomiosarcoma, which had persisted up to 7 months prior to
treatment. The patients were divided into two groups:

Group I: Seven patients where the ulcerated lesions averaged 9 cm long
and 12 cm in diameter.

Group II: Five patients where the ulcerated lesions were greater than
12 cm.times.2 cm.

The patients of each group were treated by the application to affected
skin areas of about 200 mg of petroleum jelly containing 3 wt % of
tetrasilver tetroxide three times daily over a period of 30 days.

All patients were examined daily by a dermatologist to evaluate the
effectiveness of the treatment.

Summary of Results

Group I: All patients experienced a commencement of healing of the
ulcers by day 27 from the start of the treatment. By day 30 the ulcers
were dry. On day 40 new cells had replaced infected cells. By day 45
ulcers and sores were no longer visible and new tissue was evident
replacing the ulcerated tissue.

Group II: All patients showed a commencement of the healing of the
ulcers by day 28 from the start of the treatment. By day 40 the ulcers
had almost disappeared and a biopsy confirmed that 80% of the diseased
tissue had been replaced with healthy tissue. From a clinical
standpoint, the ulcers were no longer visible.

The tests demonstrate that the topical application of tetrasilver
tetroxide is effective in curing infections and healing skin ulcers
associated with Rabdomiosarcoma, and without side effects.

Based on all of the test data described above, the healing mechanism
associated with the use of tetrasilver tetroxide to treat and cure at
least some skin diseases appears to be more than simply the killing of
pathogens and curing infections which tend to aggravate disease and
retard the natural healing process. The data indicate that healing is
brought about even in cases where no abnormal bacteria counts or
infection is evident. This suggests that tetrasilver tetroxide perhaps
also acts against autoantibodies which trigger autoimmune reactions
associated with diseased tissue.



United States Patent 5,676,977
Antelman October 14, 1997
Method of curing AIDS with tetrasilver tetroxide molecular crystal
devices

Abstract

The diamagnetic semiconducting molecular crystal tetrasilver tetroxide
(Ag.sub.4 O.sub.4) is utilized for destroying the
AIDS virus, destroying AIDS synergistic pathogens and immunity
suppressing moieties (ISM) in humans. A single intravenous
injection of the devices is all that is required for efficacy at
levels of about 40 PPM of human blood. The device molecular crystal
contains two mono and two trivalent silver ions capable of "firing"
electrons capable of electrocuting the AIDS virus, pathogens
and ISM. When administered into the bloodstream, the device electrons
will be triggered by pathogens, a proliferating virus and
ISM, and when fired will simultaneously trigger a redox chelation
mechanism resulting in divalent silver moieties which chelate
and bind active sites of the entities destroying them. The devices are
completely non-toxic. However, they put stress on the
liver causing hepatomegaly, but there is no loss of liver function.
Inventors: Antelman; Marvin S. (Rehovot, IL)
Assignee: Antelman Technologies Ltd. (Providence, RI)
Appl. No.: 08/658,955
Filed: May 31, 1996
Related U.S. Patent Documents

Application Number Filing Date Patent Number Issue Date
310859 Sep., 1994

Current U.S. Class: 424/618 ; 514/495
Current International Class: A61K 33/38 (20060101); A61K 033/38 ()
Field of Search: 424/618 514/495
References Cited [Referenced By]
U.S. Patent Documents

4415565 November 1983 Wysor
4915955 April 1990 Gomori
4952411 August 1990 Fox, Jr. et al.
5073382 December 1991 Antelman
5078902 January 1992 Antelman
5089275 February 1992 Antelman
5211855 May 1993 Antelman
5223149 June 1993 Antelman
5336499 August 1994 Antelman
5571520 November 1996 Antelman

Other References

"Is The AIDS Virus A Science Fiction?" by Peter H. Duesberg and Bryan
J. Ellison, Policy Review, Summer 1990, pp. 40-51..

Primary Examiner: Hulina; Amy
Attorney, Agent or Firm: Salter & Michaelson
Parent Case Text


This application is a continuation-in-part of patent application Ser.
No. 08/310,859 filed Sep. 22, 1994, now abandoned.
Claims


What is claimed is:

1. A method of treating AIDS-afflicted humans comprising injecting a
multitude of tetrasilver tetroxide molecular crystals into
the bloodstream of the human subject.

2. A method for increasing white blood cell counts in AIDS-afflicted
humans comprising injecting a multitude of
tetrasilver tetroxide molecular crystals into the bloodstream of the
human subject.

3. Methods of treating AIDS-affilicted humans according to claims 1-2
where the concentration of said molecular crystals is
approximately 40 PPM of the total blood weight of the human subject.
Description


BACKGROUND OF THE INVENTION

The present invention relates to the employment of molecular crystals
as anti-AIDS devices, but more particularly to the
molecular crystal semiconductor tetrasilver tetroxide Ag.sub.4 O.sub.4
which has two monovalent and two trivalent silver
ions per molecule, and which through this structural configuration
enables intermolecular electron transfer capable of killing
viruses and binding them to the resulting silver entity so that a
single intravenous injection will completely obliterate acquired
immune deficiency syndrome (AIDS) in humans. Furthermore, said devices
are capable of killing pathogens and purging the
bloodstream of immune suppressing moieties (ISM) whether or not
created by the AIDS virus (HIV); so as to restore the immune
system.

The present invention is based on concepts previously elucidated in
applicant's U.S. Pat. No. 5,336,499 which discloses the
destruction and inhibition of bacteria, algae and the AIDS virus in
nutrient life supporting systems by using said silver oxide
devices. Example 3 of said patent discloses that 18 PPM of said
crystal devices could totally suppress the AIDS virus
(page 6, line 5). Subsequent to the filing of the aforementioned
patent, further testing revealed complete 100% destruction
of the AIDS virus in vitro at 20 PPM, and the fact that said devices
were harmless when ingested and inhaled, being non-toxic.

Encouraged by these evaluations and successes, applicant obtained
permission to evaluate the crystals in vitro against
murine acquired immune deficiency syndrome (MAIDS). Only one facility
in the State of Israel is licensed for these evaluations,
namely, the Kaplan Hospital in Rehovot, Israel, which is affiliated
with the Hebrew University-Hadassah Medical School where
said evaluations were done.

The initial evaluations entailed experimenting with various silver
moieties cited in applicant's aforementioned patent,
concentrations, non-reactive buffers and modes of administration.
After about 18 months of judicious efforts and initial failures,
success was finally achieved in destroying the MAIDS virus in C57BL
mice with a single intravenous injection. The results of
this test program comprise Example 5 of U.S. Pat. No. 5,336,499. After
success with mice, the inventor was able to test the
efficacy of said devices on two select etiological groups of terminal
AIDS patients in a clinic in Tegucigalpa, Honduras,
Central America.

The AIDS patients comprised the etiological subgroups, Candidiasis and
Wasting Syndrome. Current indicator diseases for
diagnosing AIDS which have been expanded by the CDC, fall into the
following five major categories with the approximate
percent distribution among AIDS patients:

______________________________________ 1. P. carinii pneumonia 51% 2.
Wasting syndrome 19% 3. Candidiasis 13% 4.
Kaposi's sarcoma 11% 5. Dementia 6%
______________________________________

This invention concerns itself with the treatment and cure of
candidiasis and wasting syndrome AIDS patients with Tetrasil*.
These two groups account for approximately one third of AIDS cases.

Stedman's Medical Dictionary (Williams & Wilken's 26th Ed., 1995)
defines wasting syndrome "as a condition of 10% weight loss
in conjunction with diarrhea or fever . . . Associated with AIDS (p.
1744)."

OBJECTS OF THE INVENTION

The main object of the invention is to provide for a molecular scale
device of a single tetrasilver tetroxide crystalline molecule
capable of restoring the immunity of AIDS afflicted humans of the two
AIDS etiological subgroups, candidiasis and wasting
syndrome.


Another object of the invention is to provide for immunity restoration
in said AIDS afflicted humans through a single injection.

Another object of this invention is to destroy ISM in humans
manifesting AIDS diseases of said AIDS etiological subgroups
irrespective as to whether said ISM was HIV induced, since it is known
that humans may manifest AIDS and still be HIV negative,
and thus restore the immune system in said humans.

Another object of this invention is to destroy the AIDS virus when
present in the systems of said AIDS afflicted humans.

SUMMARY OF THE INVENTION

This invention relates to a molecular scale device not only capable of
destroying the AIDS virus, but of purging the human
bloodstream of pathogens and restoring immunity to AIDS patients of
the candidiasis and wasting syndrome categories.
Said molecular device consists of a single crystal of tetrasilver
tetroxide (Ag.sub.4 O.sub.4). The crystal lattice of this molecule
has a unique structure since it is a diamagnetic semiconducting
crystal containing two mono and two trivalent silver ions,
which in effect are capable of "firing" electrons under certain
conditions which will destroy AIDS viruses, other pathogens
and immune suppressing moieties (ISM), not only through the
electrocution mode, but also by a binding process which occurs
simultaneously with electron firing, namely, binding and chelation of
divalent silver, i.e., the resulting product of the electron
transfer redox that occur when the monovalent silver ions are oxidized
and the trivalent ions are reduced in the crystal. The binding/
chelation effect occurs at active sites of the AIDS virus, pathogens
and ISM. Because of the extremely minute size of a
single molecule of this crystal, several million of these devices may
be employed in concert to destroy a virus colony to purge a
life support system of ISM and pathogens with the consumption of only
parts per trillion of the crystal devices. Thus an
optimum of 40 PPM of the devices by weight of human blood was found to
be sufficient to completely obliterate AIDS. This
concentration is slightly over double of the optimum concentration
recommended in applicant's aforementioned U.S. patent
for the destruction of the human AIDS virus in vitro. Other details
concerning the structure of the crystal and its mechanism
against pathogens, the AIDS virus and ISM would analogously hold here,
and have already been further elucidated in said
patent.

The actual destruction of pathogens, ISM and the AIDS virus is
effectuated by injection of a suspension of these devices in
distilled or deionized water with a non-reacting electrolyte directly,
i.e. intravenously, into the bloodstream. A single injection
is all that is required under these conditions. Accordingly, humans
injected in this manner, upon being inspected after three
weeks or more had elapsed and compared with similar humans that had
been given placebos, were completely cured of AIDS.
The control group still manifested AIDS. Accordingly, the tetrasilver
tetroxide device performed in concert with and in full
conformity with the ultimate objects of this invention. Furthermore,
three out of four wasting syndrome terminal patients and
four out of the five candidiasis terminal patients were still alive in
1995 after a year and a half had elapsed from their initial
injection. By that time all the AIDS patients had been released from
the clinic and allowed to return home.

Other objects and features of the present invention shall become
apparent to those skilled in the art when the present invention
is considered in view of the accompanying examples. It should, of
course, be recognized that the accompanying examples
illustrate preferred embodiments of the present invention and are not
intended as a means of defining the limits and scope of
the present invention.

EXAMPLE 1

Five patients afflicted with AIDS of the candidiasis etiological
category were segregated for Tetrasil treatment. The rationale
For selecting them was based on facts presented in an article by Peter
H. Duesberg and Brian J. Ellison entitled
"Is The AIDS Virus A Science Fiction?" (Policy Review, Summer 1990 pp.
40-51). Only the factual presentations of the
article were utilized and the hypothesis of the authors was ignored.
The facts presented in the article related to the
method of selecting AIDS patients based on the five aforementioned
etiological subgroups targeted by the CDC, and the
evidence presented, that there is AIDS without HIV as well as with it
so that an anti-viral agent in most instances will not
necessarily restore the immunity system.

Evaluations with Tetrasil were conducted on AIDS patients at Lucha
Contra el Sida, Comayaguela, Honduras.
The patients two weeks prior to inoculation were removed from their
AZT, AIDS therapy. Tetrasil was administered
at approximately 40 PPM of blood volume per patient as a suspension in
a proprietary buffer solution (pH=6.5), supplied
by Holipharm Corporation.

The results of evaluations with candidiasis are tabulated in Table I
under its disease category. All patients evaluated were
terminal. Some, however, were in moderate (m) condition and others in
poor (p) as designated in the Table. The I and F
designations refer to initial and final values as shown. WBC indicates
white cell blood count. The H column, following CD 8,
indicates whether hepatomegaly occurred. This was an unfortunate
consequence of the treatment which resulted in enlarged
livers in all patients except the second one. Despite hepatomegaly,
there was no interference with liver function.

The onset of hepatomegaly was not spontaneous and varied from patient
to patient, being in the range of 4-16 days.

It should also be noted that shortly after injection of Tetrasil there
were indications of fever
(symbolized by T in the Ag.sub.4 O.sub.4 column), sometimes
accompanied by fatigue (F). The body temperature was invariably
38.5.degree. C. (101.3.degree. F.). This was indicative of restoration
of the immune response of the body, since normally
the body will destroy pathogens when the immune system is functional
by raising the temperature. The patient who died;
first responded favorably to Diflucan, which previously gave no
response. He was cured of his candidiasis, but unfortunately
succumbed to his previous body damage. All the other candidiasis
syndrome people who previously did not respond to the
indicated medications subsequently responded after the Tetrasil
treatment. Further evidence of the recovery of the AIDS
patients manifested itself 30 days after the initial injection when
white blood cell counts were taken. They are shown in Table
I under the WBC column, which gives the initial and final WBC. All
candidiasis patients showed a dramatic increase in their
white blood cell counts, indicative of the restoration of their
immunity systems.

EXAMPLE 2

The above protocol of Example 1 was repeated with AIDS patients
exhibiting wasting syndrome. The results of their treatment are
tabulated in Table I under the disease category of said syndrome. It
should be noted that two of the four wasting syndrome patients showed
improved white blood counts. The female patient, whose condition
improved from poor and terminal to be
among the living, showed a decrease in the WBC. However, she showed an
increase in body temperature which was indicative of immune response.
The test results indicate that one cannot rely on a single factor to
indicate the demise of AIDS.
The usual HIV marker CD 4 initial and final are irrelevant. ISM
suppression appears to be more critical than the destruction of HIV.
AIDS was suppressed, any permanent damage that had been done to the
patients in the course of their succumbing to AIDS was not obviously
cured or corrected by said crystal device treatment, rather said
injury persisted and the patient was improved with respect to AIDS but
still suffered from said permanent injury or impairment previously
inflicted.

TABLE I
__________________________________________________________________________
Response of AIDS Patients to
Single 40 PPM Ag.sub.4 O.sub.4 Inoculation Date Weight DISEASE
PATIENT Inoc. WBC CD 4 DEATH Lbs. Group Sex Age
Medictn 1994 I F I F CD 8 H 1944 I F Ag.sub.4 O.sub.4
__________________________________________________________________________
Candidiasis M p 28 Diflucan 5/5 1,200
4,200 41 -- 221 + 6/11 82 76 T F m 33 " 5/5 6,000 6,700 554 872 394 -
98 98 T F m 33 Ketaconzl 5/27 2,600 3,850 248 181 951
+ 123 123 T M p 62 " 6/2 3,300 3,700 89 237 59 + 105 92 F F m 31
Pentamidn 6/2 2,400 3,050 9 181 65 + 121 118 Pain Wasting
M m 27 5/27 3,600 4,600 39 14 709 + 119 120 T Syndrome M m 28 5/27
2,750 -- 10 -- 60 + 7/19 121 119 T, F F p 43 5/27 3,600
2,700 68 246 248 + 101 98 T, F M m 19 5/10 3,850 5,400 137 36 48 + 103
106 T, F
__________________________________________________________________________

As this invention may be embodied in several forms without departing
from the spirit or essential characteristics thereof, the present
embodiments are therefore illustrative and not restrictive, since the
scope of the invention is defined by the appended claims rather than
by the description preceding them, and all changes that fall within
the metes and bounds of the claims or that form their functional as
well as conjointly cooperative equivalents, are therefore intended to
be embraced
by these claims.


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