Problems of Biological Medical and Pharmaceutical Chemistry

According to their chemical structure, chlorophylls are magnesium complexes of various tetrapyrroles, the main plant sources of which are dioecious nettle, alfalfa, mulberry, eucalyptus, various types of algae and common spruce. Chlorophylls A, B, C, and D have been studied, but as a percentage, species A and B. Chlorophylls have high biological activity, which is why they are of interest for the development of medicines. Currently, chlorophyll derivatives are actively used for photodynamic therapy of cancer diseases. Proven pharmaceutical effects include hematopoietic, wound healing, bactericidal, antiviral, and immunostimulating.

The purpose of the review is to systematize information sources on the chemical structure, properties, biological activity, technology and standardization of chlorophylls isolated from plant raw materials.

Information about chlorophylls is summarized, their chemical structure, plant sources, properties and biological activity are studied. Special attention is paid to extraction methods and quantitative determination of chlorophylls in plant raw materials. It has been established that the main methods of chlorophyll extraction from plant raw materials are maceration with ethyl alcohol. To intensify the release of chlorophylls, an ultrasonic bath and a microwave oven are used, as well as methods of extraction with a supercritical liquid, deep eutectic solvents, using an accelerated dissolution extractor. Methods of spectrophotometry, photometry, inductively coupled plasma with optical emission spectrometry, nuclear magnetic resonance (NMR) spectroscopy, and high performance liquid chromatography (HPLC) are used to quantify chlorophylls in the obtained extracts. Currently, chlorophylls are mainly used as natural pigments. In this review, they are considered from the point of view of their use in medicine as an active pharmaceutical substance (APS). The state register of medicines and the pharmaceutical market have been studied. It has been established that in Russia, mainly medicinal products for external and topical use are registered with chlorophylls. Preparations with chlorophylls from eucalyptus and dietary supplements have been registered for internal use.

Introduction. Nifedipine, a typical representative of 1,4-dihydropyridine, is widely used in the treatment of various diseases due to its vasodilatory and antihypertensive effects. Low water solubility is a common problem for drugs such as nifedipine, limiting their bioavailability and effectiveness. This leads to a decrease in the therapeutic effect and the need to increase the dosage, which, in turn, can increase the risk of side effects. Nanotechnology offers a promising approach to overcome this barrier, allowing the creation of dosage forms with improved pharmacokinetic properties.

Aim of the study. Developing a method for increasing the solubility of nifedipine by obtaining nanoparticles using a high-energy ball mill.

Material and methods. The following were used in the work: nifedipine (Tamico, Syria), methanol (Biosolve Chimie, France), potassium dihydrogen phosphate, sodium heptanesulfonate and orthophosphoric acid (85%) (Isolab, Germany), purified water and Tween 20 (Merck, Germany). Nifedipine was ground using a high-energy ball mill (Retsch PM 400). During the research, the dissolution profile of nifedipine and nifedipine nanoparticles was studied by examining their saturated solution using HPLC. To characterize the sizes of the ground samples, they were examined using scanning electron microscopy (SEM).

Results. The use of a high-energy ball mill (Retsch PM 400) allowed us to obtain nifedipine nanoparticles with significantly improved solubility characteristics. It was found that the solubility of nifedipine nanoparticles was 9.05%, which is 38 times higher than that of the original substance (0.24%).

Conclusions. The findings of this study highlight the transformative potential of high-energy ball milling in enhancing the solubility of nifedipine, a crucial component in pharmaceutical formulations. By employing this innovative approach, we successfully produced nifedipine nanoparticles with demonstrably improved solubility compared to the conventional powder form. This achievement opens a new frontier in the development of nifedipine-based dosage forms with enhanced bioavailability. The increased solubility of nifedipine nanoparticles signifies a promising avenue for optimizing drug delivery and potentially improving therapeutic efficacy.

Introduction. Rabies is an infectious disease with absolute mortality. Currently, there are no effective drugs and treatment regimens for people with signs of rabies. The only strategy for post-exposure rabies prevention is vaccination and administration of rabies immunoglobulin. At the same time, the search for new treatment and prevention regimens for the disease remains relevant, which is why the development and study of antiviral drugs effective against the rabies virus is relevant.

Objective of the study. To study the antiviral activity of a drug based on single-stranded high-polymer RNA of Sacharomyces cerevisiae, which has previously shown antiviral activity against other pathogens of viral infections.

Material and methods. The work investigated the effect of the drug based on single-stranded high-polymer RNA of S. cerevisiae "Amphievovir" on the level of reproduction of the attenuated rabies virus strain "Saratov" on the Vero cell culture model.

Results. The studied drug in the concentration range from 10 to 20 μg/ml and added to the cell culture no later than 24 hours after infection reduces the titer of the virus by 24.6 to 36.2% compared to the control group. Adding the drug to the infected cell culture later than 24 hours reduces its antiviral effect.

Conclusions. The studied drug based on single-stranded high-polymer RNA of S. cerevisiae has antiviral activity against the rabies virus in vitro. The results allow us to justify further study of post-exposure rabies prophylaxis regimens using drugs based on immunostimulating and immunomodulatory RNA.

Introduction. Wharton's jelly of the human umbilical cord is a connective tissue of extraembryonic origin that maintains characteristics of the embryonic phenotype, including the capacity for rapid tissue regeneration and scar-free healing of fetal wounds. Decellularization refers to the removal of cells and cellular components from biological tissues while preserving the essential structural and compositional features of the extracellular matrix.

The aim of the work was to evaluate the structural and biological characteristics of the decellularized extracellular matrix from Wharton's jelly of the human umbilical cord.

Material and methods. Decellularization was performed by a detergent method using a sterile solution of sodium dodecyl sulfate at a concentration of 0.01% for 24 h at room temperature. The component composition of Wharton's jelly of the human umbilical cord before and after the decellularization process was assessed using spectral analysis methods. To study the biological characteristics of the decellularized extracellular matrix from Wharton's jelly of the human umbilical cord, the MTT test and the subcutaneous implantation model in mice were used.

Results. The content of total collagen by hydroxyproline in Wharton's jelly of the human umbilical cord before decellularization ranged from 244.8 to 507.2 μg/mg, and after – from 398.9 to 777.3 μg/mg, hyaluronic acid and sulfated glycosaminoglycans – from 11.5 to 16.5 and from 16.1 to 22.5 μg/mg before decellularization and from 15.6 to 22.1 and from 25.6 to 29.6 μg/mg after decellularization, respectively. Multiple collagen types (I, III, IV, V, VI, XII), as well as fibronectin, lumican, decorin, biglycan, and tenascin were identified in Wharton's jelly of the human umbilical cord. The absence of cytotoxicity of model media based on extracts from decellularized extracellular matrix from Wharton's jelly of the human umbilical cord was revealed. No signs of rejection or enhanced cellular inflammatory response were observed during subcutaneous implantation in mice.

Conclusions. The practical possibility of developing a drug and/or medical device for regenerative medicine based on decellularized extracellular matrix from Wharton's jelly of the human umbilical cord was demonstrated.

Introduction. Carbonylation, as a variant of oxidative modification of proteins, serves as an indicator of tissue damage during oxidative stress, including that caused by hypoxia. Some regulators of the metabolic response to oxygen deficiency, such as the NO donor arginine and the mitochondrial metabolite succinate, are also able to influence the modulation of cellular antioxidant defense and oxidative stress.

The aim of the study was to evaluate the effect of arginine and succinate in combination on the degree of protein carbonylation and antioxidant status during hypoxia.

Material and methods. The study included 40 wistar rats divided into 5 groups (n=8): animals exposed to normobaric chronic hypoxia while being kept in a hermetic chamber until the oxygen level decreased to 10% once a day for 14 days (1); receiving arginine at a dose of 500 mg/kg b.w. per day for 10 days (2); succinate at a dose of 100 mg/kg b.w. per day for 14 days (3); receiving arginine and succinate (4); exposed to hypoxia and receiving arginine and succinate (5). In the mitochondria of the seminal vesicles and epididymis, the activity of superoxide dismutase was assessed by inhibition of the reaction with quercetin, the degree of protein carbonylation by the reaction with 2,4-dinitrophenolhydrazine, from which the reserve-adaptive oxidative modification potential of proteins was calculated. The Mann-Whitney criterion was used for statistical analysis.

Results. The combination of arginine and succinate resulted in an increase in the degree of protein carbonylation and a decrease in the reserve-adaptive oxidative modification of mitochondrial proteins of the seminal vesicles and epididymis, while the superoxide dismutase activity remained at the level of group 2, which is lower than in group 3. The complex effect of the studied metabolites under hypoxia had a protective effect on mitochondrial proteins, reducing the degree of their oxidative modification, which is especially true for mitochondrial proteins of the epididymis head.

Conclusions. Under conditions of physiological NO synthesis, succinate helps maintain superoxide dismutase activity, which does not occur under hypoxia-like conditions. Activation of NO synthesis by arginine in a complex with succinate is an effective way to correct the oxidative modification of mitochondrial proteins under hypoxia.

Introduction. Sequoia sempervirens (D.Don) Endl. are the tallest long–lived relict plants capable of accumulating unique secondary metabolites that can be used in phytopharmacognosy. To preserve the biodiversity of rare plant species, it is advisable to apply biotechnology methods and create in vitro genetic banks, including by the method of clonal micropropagation. Stress-resistant and highly productive plants can be created using methods of cellular biotechnology, in particular, in vitro cell selection.

The purpose of the study. development of technology for in vitro production of highly productive microclones of relict gymnosperms of the genus Sequoia, study of the formation and localization of secondary metabolites and their biological activity.

Material and methods. The object of the study was Sequoia sempervirens (D.Don) Endl. plants. Aseptic culture was obtained from shoots of the first year of vegetation of intact plants. Explants were cultured on MS nutrient medium with different hormonal composition. The localization of phenolic compounds was studied in leaves, stems, apical buds of intact sequoia plants and in microclones, as well as in callus tissue. Histochemical methods were used for this: the material was stained with 0.08% Fast Blue reagent raster for the sum of phenolic compounds, and a reaction with vanillin reagent in hydrochloric acid vapor was used to study the localization of flavans (catechins and proanthocyanidins). The quantitative content of different classes of polyphenols was determined using spectrophotometric methods. The cytotoxic properties of the extracts were studied using an MTT test.

Results. Microclone cultures of evergreen sequoia (Sequoia sempervirens (D.Don) Endl.) were obtained in vitro. The dependence of culture growth in vitro on the hormonal composition of the nutrient medium and the endogenous content of polyphenols in the primary explant has been studied. The accumulation and localization of secondary compounds in in vitro Sequoia sempervirens microclones has been shown for the first time. Secondary metabolites are predominantly localized in epidermal, parenchymal and conductive tissues in both intact plants and microclones in vitro. The localization of secondary compounds in in vitro cultures as producers of substances with high biological activity is characterized. Studies on the cytotoxicity of extracts of sequoia cultures in vitro demonstrate high rates in relation to the HeLa cervical cancer cell line and the human glioblastoma A172 line.