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All of these aspects are studied in radiobiology [1], a branch of biology closely related to physics. Applied to the medical field, radiobiology examines the effects of ionizing radiation (produced by radionuclides or charged particle beams) on cells and tissues, aiming to optimize dose administration, and minimize risks while maximizing benefits. As well as studying the internalization and externalization kinetics of drugs within cells, the mechanism of activation of specific biochemical pathways, as well as changes in cellular morphology, are also explored in this research. Furthermore, the creation of a standard in vitro model, using cells, is essential for correlating dose-related effects and biological endpoints, in order to highlight the risks and benefits of radioactivity exposure to radiopharmaceuticals.
The concept of radiotheranostics within the framework of treatments used in case of cancer disease. In this case, the objective of the research is to develop pharmaceutical formulations that keep the radionuclide component unchanged, but adapt the ligand component according to its (a) diagnostic or (b) therapeutic purpose.
Considering biologists and biotechnologists as the target audience, this publication has been structured in such a way as to offer an ordered overview of the various basic issues useful for understanding this review, in order to guide the reader to a multidisciplinary path, from imaging techniques to radiotheranostics [3], which are closely related to nuclear medicine, radiopharmaceutical radiobiology [9], physics [10], radiochemistry [11], artificial intelligence (machine learning and deep learning), radiomics techniques [12], and radiogenomics [13]. The reader will be directed to various works in which the use of a γ-counter for in vitro radiometric assays is considered. These assays are used for the evaluation of radioactivity in some cell lines. It should be noted that the radiometric measurements and in vitro tests referred to throughout the review do not take into consideration the quality control understood as Good Manufacturing Practice (GMP), but aim to introduce the case for radioactivity measurement using radiometric counters and two-dimensional cell cultures treated with radiopharmaceuticals, that can be carried out in carefully equipped research laboratories with radiation protection.
Two primary components characterize radiopharmaceuticals: (i) a radionuclide that confers tracing or damaging properties to the molecule by ionizing radiation, and (ii) a specific biomolecule or ligand defining the radiopharmaceutical binding and uptake of cell targets, as well as two additional significant components, a chelator and a linker, which ensure drug chemical stability (see Figure 2). Contrary to radiopharmaceuticals containing radionuclides such as iodine, fluorine, or carbon isotopes that are covalently attached to carriers, most imaging radiopharmaceuticals contain a metal as part of their design. As a result of the presence of the metal, high thermodynamic, and kinetic stability is required to avoid transmetallation reactions, i.e., the metal binds to other parts, decreasing selectivity and increasing background noise (and radio-toxicity). Radionuclides with long half-lives may also cause biological damage. It is also important to limit the production of metabolites, especially those labeled with radionuclides. Metal radionuclides require knowledge of the coordination chemistry of their metal ions to optimize their choice of ligands from the perspective of their chemistry [17]. The chelator and linker are often combined into one molecule, known as a bifunctional chelator. The bifunctional chelator is attached to the vector by a linker or spacer, and is a molecule that contains the appropriate donor groups for the formation of a coordination compound with the radionuclide. Radiometals are the only ones that apply to this concept, and to ensure that no interference with the interaction with the target is caused by the coordination of the metal, the linker is positioned separately from the bioactive part of the molecule.
The design of a canonical radiopharmaceutical compound. A radiometal represents a specific radionuclide. BFC stands for a chemical compound that chelates radionuclides; a linker/spacer is a link between two molecules; the tumor-targeting vector recognizes and binds a tumor biomarker to a cell.
Innovative radionuclides and chelators are increasingly in demand in nuclear medicine. This is because the physical characteristics of radioisotopes and the biological characteristics of compounds are not always compatible with each other. A number of radionuclides, in fact, have a long half-life compared to targeting vectors (e.g., antibody fragments, small-molecule peptides that bind specific membrane receptors that are abundant on the surface of target tumor cells), which makes radioactive labeling with these radioisotopes incompatible [18]. Therefore, the goal is to find radioisotopes with a half-life compatible with the biomolecule carrier and optimize a chelator that is chemically compatible with the radionuclide to stabilize the radiopharmaceutical [19]. A major challenge in optimizing novel radiopharmaceuticals concerns the decay time and the limited use of some radioisotopes used in some assays [20]. This concerns the method of administration of these radionuclides for SPECT diagnostic use, such as, for example, 67Ga (half-life time 78 h), which is administered as Ga3+ citrate, which hydrolyzes slowly and is transported in the body by transferrin [21]. The long half-life of 67Ga allows its use even at very long distances from the place of production for the imaging of inflammatory processes and tumors. The use of the 68Ga isotope is sometimes preferred in the PET technique. This is because on the one hand, it uses the same 3+ ion family chemistry that they have as the only stable oxidation state Ga3+ and forms more stable structures with smaller macrocycles, type NOTA, than with DOTA. On the other hand, it also has the advantage of having a much shorter lifetime of about 68 min [22]. Thus, the goal is to develop radiopharmaceuticals that have radioisotope decay times comparable to biological half-lives [19].
Depending on other parameters, such as the depth of the target tissue and the resolution of the PET image, the energy of the particles emitted by each radionuclide can vary [23]. Optimal image quality depends on the energy of the particles emitted [24]. Table 1 shows the physical characteristics of all the radionuclides mentioned in this manuscript. Moreover, the goal of radionuclide production is to achieve maximum quality and purity of the radioisotope, which is directly connected with the safety profile and, therefore, image quality [25].
In radiotheranostics, the use of one radioisotope or a pair of radionuclides to mark the same targeting vector for a dual diagnostic and therapeutic purpose, is a hot research topic, aiming to optimize the use of a single drugs that perform two distinct functions, visualization and treatment through the exchange of specific radionuclides [26]. A theranostic pair may consist of two isotopes of the same element or two different elements with similar chemical characteristics. In the latter case, we report the use of 68Ga for PET or 111In for SPECT [27], followed by radionuclide therapy with lutetium-177 (177Lu) [28], yttrium-90 (90Y) [29] or actinium-225 (225Ac) [30]. Copper-64/copper-67 (64Cu/67Cu) [31] is an example of a radiotheranostic pair involving two isotopes of the same element, along with iodine-123-124/iodine-131 (123/124I/131I) [3], yttrium-86/yttrium-90 (86Y/90Y) [26], terbium isotopes [27], and scandium isotopes as well.
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