Problems of Biological Medical and Pharmaceutical Chemistry

The topic of using plants in the treatment of various diseases is not new.  Since ancient times, people have accumulated knowledge and skills of their use and widely used them for health improvement. Moreover, both monoplants and their complexes in the form of herbal preparations were used. The latter was often more preferable, since it is believed that the multicomponent effect of plants on the body is more effective. However, despite this, herbal preparations currently account for about 20% of herbal medicines, which cannot be considered justified. This is particularly true for disen mixtures, which have a number of advantages over charges produced in bundles (in bulk).

This article discusses the current state of development, research and release of dosen mixtures of herbal medicines.

This review is devoted to the study of the chemical composition and pharmacological activity of the plant – Amorpha fruticosa L., growing in the Astrakhan region. According to scientific literature, it was revealed that A. fruticosa contains a unique complex of biologically active compounds, and various extracts based on it exhibit antioxidant, antidiabetic, cytotoxic, antimicrobial, anti-inflammatory, antitumor activity. Analysis of the results of modern studies indicates the uniqueness of the chemical, physiological and therapeutic properties of this plant. This creates real prerequisites for the scientifically substantiated use of Amorpha as a means of preventing and treating metabolic, oncological and many other diseases, the pathogenetic mechanisms of which include changes in the antioxidant system, the development of oxidative stress, and inflammatory processes.

Introduction. Despite the wide distribution and extensive use of Artemisia L. species in official and traditional medicine for the treatment of inflammatory, infectious, and other diseases caused by fungi, bacteria, and viruses, the chemical composition of Artemisia messerschmidtiana Bess. growing in the territory of Buryatia remains insufficiently studied.

Aim of the study. To perform a pharmacognostic analysis of the aerial part of A. messerschmidtiana.

Materials and Methods. The object of the study was the whole aerial part of A. messerschmidtiana collected during the flowering stage near Udinsk village, Khorinsky District, Republic of Buryatia (August 2023). Anatomical and diagnostic features, as well as the main quality indicators of the raw material, were determined according to the methods of the State Pharmacopoeia, 15th edition. The essential oil was obtained by steam distillation. Lipid fractions were isolated using a modified Bligh and Dyer method followed by acid methanolysis. The chemical composition of the essential oil and lipid fraction was analyzed using gas chromatography–mass spectrometry (GC–MS).

Results. The main anatomical and diagnostic characteristics of the raw material were determined. The leaf epidermis consists of isodiametric cells with slightly sinuous walls; stomata are of the anomocytic type, mainly located on the lower epidermis. T-shaped trichomes and oval, tiered essential oil glands are characteristic features. The main quality indicators were as follows: moisture content 7.6%, total ash 7.9%, acid-insoluble ash 0.6%, ethanol-soluble extractives (70%) 42.8%, organic impurities 0.8%, browned and blackened parts 2.0%. The yield of essential oil was 0.3%, its main components were 1,8-cineole (11.6%), camphor (19.6%), borneol (4.0%), α-terpinene (6.1%), isoterpinolene (3.4%), phytol (7.0%), dehydrosesquicineole (4.8%), and caryophyllene oxide (2.3%). The yield of the lipid fraction was 4.5%, containing ten higher fatty acids - palmitic (42.0%), linoleic (16.6%), linolenic (14.9%).

Conclusion. The main anatomical and diagnostic features and quality indicators of A. messerschmidtiana were established. The composition of the essential oil was found to be stable regardless of the growing region or extraction method. The dominant compounds are monoterpenoids – 1,8-cineole, camphor, and borneol, while caryophyllene oxide is the main sesquiterpenoid. The lipid fraction is characterized by palmitic, linoleic, and linolenic acids. The findings confirm the potential of A. messerschmidtiana as a promising medicinal plant source for the development of new herbal medicinal preparations.

This article provides an overview of platinum-based anticancer agents. Drugs based on coordination complexes of divalent platinum Pt(II) and tetravalent platinum Pt(IV) are widely used in chemotherapeutic treatment for malignant neoplasms. The development and mechanism of action of cisplatin and its derivatives, which have been approved for clinical use, are discussed. To overcome the disadvantages of Pt (II) drugs, prodrugs based on Pt (IV) have been proposed. Approaches to modifying Pt (II) and Pt(IV) drugs to increase their selectivity for tumor cells are being discussed for future use in photodynamic therapy and photoactivated chemotherapy. The photophysical properties of platinum complexes with photosensitizers suggest the possibility of a dual effect on cancer cells. Based on the analysis of available literature data, we conclude that, despite the achieved results, further development of methods to reduce toxicity and increase selectivity while maintaining therapeutic effectiveness of drugs is necessary. Additionally, an expansion of the range of action is required. We consider combined methods of cancer treatment using platinum-based drugs, which may lead to a synergistic effect, as promising approaches.

Introduction. Long non-coding RNAs (LncRNAs) are transcripts whose role in carcinogenesis is poorly understood. LncRNA expression determines the biological properties of a cell, including invasive potential and ability to migrate. Methylation status changes lncRNA functions and can be a marker of tumor prognosis.

The aim of the work is study the methylation frequency of the LncRNA genes GAS5, ZEB1-AS1 in breast cancer and to evaluate the clinical significance.

Material and methods. 140 patients with breast cancer were examined, mainly with T1 – (38.7%) and T2 – (55.6%) stages, N0 status – (54.0%), N1 status – (37.9%). Luminal type of breast cancer – 80% (n = 113), non-luminal – 20% (n = 27). Morphologically: invasive ductal carcinoma (77.1%), G2 differentiation (62.9%) were predominantly detected. Methylation of CpG regions of promoter regions of the GAS5, ZEB1-AS1 lncRNAs genes in paired tumor and histologically unaffected breast tissue samples was studied using the methyl-specific PCR method. The T100 amplifier (Bio-Rad, USA) and Lasergene 17.1 primers from DNASTAR (USA) were used in the work. Statistical data processing was performed in the SPSS, v. 23 program.

Results. The frequency of methylation of the GAS5, ZEB1-AS1 genes in the tumor is higher than in histologically unaffected breast tissue. Gene methylation is associated with age, R = 0.43 (GAS5) and R = 0.26 (ZEB1-AS1), and increases the stage tumor process. A correlation was established between GAS5 gene methylation and the T index (R = 0.21, p < 0.05), N status (R = 0.31, p < 0.05). There is an inverse correlation with the biological subtype of luminal cancer (R = –0.29, p < 0.05). The absence of methylation of the GAS5 and ZEB1-AS1 genes is associated with a decrease in 5-year overall survival and 10-year progression-free survival in breast cancer.

Conclusions. In breast cancer, methylation of the lncRNAs GAS5, ZEB1-AS1 genes in the tumor exceeds that in the unchanged breast tissue. The methylation status of the genes GAS5, ZEB1-AS1 is associated with the stage, tumor size, regional metastasis, and does not affect the long-term treatment results.

Introduction. In the context of the ongoing COVID-19 pandemic, studying all aspects of the impact of SARS-CoV-2 on the human body remains a priority. This article investigates changes in the oral fluid of patients with COVID-19, focusing on the relationship between pro-inflammatory cytokines and metabolic alterations.

Objective: To study the biochemical composition of oral fluid in COVID-19 patients and to identify the relationship between pro-inflammatory cytokines and potential metabolic changes.

Material and Methods. The study was conducted at the Samara State Medical University. Oral fluid from 157 COVID-19 patients (study group) and 89 healthy individuals (control group) was used as the research material. The following were determined in oral fluid: total protein content, albumin, C-reactive protein, metabolites (urea, uric acid, glucose, lactate), electrolytes, cytokines (interleukin-6, interleukin-8), and enzyme activity (alanine aminotransferase, aspartate aminotransferase, creatine phosphokinase, gamma-glutamyl transpeptidase, alkaline phosphatase, lactate dehydrogenase). Statistical processing of the data was performed using the StatTech v. 4.8.7 software (developer – Stattech LLC, Russia).

Results. The study evaluated metabolic and immunological changes in the oral fluid of patients with COVID-19. A significant increase in the content of total protein, sodium, chlorine, calcium, magnesium, iron, as well as interleukins-6 and -8, C-reactive protein, and activity of creatine phosphokinase, gamma-glutamyl transpeptidase, and lactate dehydrogenase was revealed. An inverse correlation was established between the level of IL-6 and the activity of alanine aminotransferase, aspartate aminotransferase, creatine phosphokinase, concentration of urea and magnesium, which may indicate alternative mechanisms of COVID-19 pathogenesis in the oral cavity. A direct correlation was found between C-reactive protein and uric acid, likely associated with oxidative stress. IL-8 did not show significant relationships with the metabolic profile.

Conclusion. The results described above highlight the relationship between the immune response, metabolic status, and local processes occurring in the oral cavity. The identified patterns necessitate further in-depth research to study the molecular mechanisms of these relationships in detail. The knowledge gained can contribute to the search for potential biomarkers to assess disease severity, predict its outcome, and monitor the effectiveness of therapy, as well as to develop new therapeutic approaches aimed at modulating immune and metabolic disorders associated with COVID-19.

Introduction. The main principle of correction of ischemic conditions of the brain is to provide antioxidant protection of neural structures through the use of antioxidants in complex pathophysiological therapy. In the research and development of such products, much attention has recently been paid to medicinal plants as sources of safe antioxidant agents, mainly from the class of flavonoids.

The purpose of the work. Determination of the cerebroprotective activity of the amount of the substance of sophoricoside (SSPH), isolated from Styphnolobium japonicum fruits to assess the prospects for its use in ischemic brain disorders.

Material and methods. The experiments were performed on mature male Wistar rats. The animals were divided into four groups. Group 1 (control) included rats with cerebral ischemia who received purified water (without treatment); groups 2, 3, and 4 included animals who received SSPH per os once for 7 days before the experiment at doses of 20, 40, and 80 mg/kg, respectively. Cerebral ischemia was reproduced by occlusion of both common carotid arteries. The cerebroprotective activity of SSPH was assessed according to the standard scale of neurological deficit.

Results. In the control group, the neurological deficit was 9,2 points, and the survival rate was 20%. With preventive course administration to rats of groups 2, 3 and 4 of the studied substance at doses of 20, 40 and 80 mg/kg, the indicators of neurological deficit were lower compared to the control by 13–22%, and the survival rate was 50, 50 and 62.5%, respectively.  In in vitro tests, SSPH inhibited DPPH^●– and OH^●– radicals, with 50% inhibition rates of IC₅₀=1,15±0,25 mg/ml and IC₅₀=301,1±15,0 mcg/ml, respectively. SSPH in concentrations of 0,1–100 micrograms/ml showed membrane-stabilizing activity in the model of peroxide/osmotic hemolysis: the indicators were 3-7%, against 100% hemolysis in the control.

The content of the main substance in the test substance was 90.2%. By HPLC-UV-MS, impurities of other flavonoids were identified in the SSPH composition: kaempferol-rhamnoglucoside, kaempferol-glucoramnoglucoside, kaempferol-diglucoside, rutin, ginestin, genistein-glucoramnoside, genistein.

Conclusions. The cerebroprotective effect of SSPH has been established. The cerebroprotective effect of SSPH has been established.  The mechanism of action of the substance under study is presumably based on the antioxidant and antiradical activity of sophoricoside, as its main component, as well as concomitant flavonoids.

Introduction. Acetylcholinesterase is a key enzyme of the cholinergic system that catalyzes the hydrolysis of acetylcholine into choline and acetate. This enzyme is found in the brain tissues and red blood cells of mammals. Studying its activity in membrane preparations of erythrocytes is relevant for diagnosing the effects of organophosphorus compounds and for developing methods to correct neurodegenerative diseases. The cytoskeleton of erythrocytes, which is formed by a spectrin–actin complex, plays an important role in maintaining the cell’s mechanical properties. The effect of cytoskeletal components on acetylcholinesterase activity provides a deeper understanding of this enzyme’s regulation.

Aim. We investigated how the cytoskeleton influences the activity and kinetic parameters of acetylcholinesterase in rat erythrocytes.

Material and Methods. Experiments were carried out on Wistar rats weighing 200–230 g, kept on a standard diet. Erythrocytes were isolated and subjected to hypoosmotic hemolysis to prepare ghosts (membrane preparations) and spectrin-free vesicles by removing part of the cytoskeletal proteins. Acetylcholinesterase activity was evaluated by the Elman et. al., (1961) method, expressed in micromoles of hydrolyzed acetylthiocholine. Kinetic parameters (K_m and V_max) were determined using the Lineweaver–Burk method.

Results. The enzyme activity in spectrin-free vesicles decreased by approximately 30 percent, although subsequent treatment with Triton X-100 restored the activity to levels comparable to those in erythrocyte ghosts. Analysis showed that the absence of the cytoskeleton did not affect K_m or V_max values, which confirms the stability of the enzyme’s catalytic properties. A reduction in protein content in the vesicles was verified by the Lowry method.

Conclusions. Acetylcholinesterase retains its functional activity after partial removal of the cytoskeleton. Treatment with detergent reveals a latent portion of the enzyme’s activity, which is associated with altered membrane orientation during vesiculation. These findings broaden the possibilities of using erythrocyte membrane preparations to model cholinesterase function under various physiological and pathological conditions.