Research Results in Pharmacology

Ascorbic acid-containing compound efficacy in ischemic brain damage

Mon, 09 Sep 2024 00:00:00 +0300

Introduction: Ischemic brain injury remains one of the main causes of disability and mortality worldwide. Protection of cellular population, depriving from oxygen supply and nutrients, is of extreme importance for further both clinical and health outcomes of timely implemented intravascular intervention. The aim: to assess anti-ischemic activity of 3-hydroxypyridine ascorbate in the in vitro and in vivo models of brain cell and tissue response to ischemia and reoxygenation.

Materials and Methods: 3-hydroxypyridine ascorbate (laboratory code 3-EA) was assessed as chemical substance (purity 99.8%) diluted in sterile phosphate-buffered saline. Intracellular Ca²⁺ response to glutamate excitotoxicity (GluTox), ischemia and reoxygenation as well as cellular viability was evaluated on NMRI murine fresh cortical neuro-glial cell culture incubated with 2-EA by registering intracellular Fura-2 and propidium iodide fluorescence respectively. Expression of apoptosis regulating genes BCL-2, STAT3, SOCS3, inflammation regulating genes TRAIL, MLKL, Cas-1, Cas-3, IL-1β и TNFα, and genes MAO-A and MAO-B was determined by real-time PCR. The substance neuroprotection was studied in male Sprague-Dawley rats with intraluminal middle cerebral artery (MCA) occlusion/reperfusion treated with 18 mg/kg of 2-EA along with neurological deficiency evaluation and morphological assessment of brain sections.

Results: Preincubation of cortical cells with 10-100 μM of 3-EA leads to inhibition of [Ca²⁺]i in cytosol of neurons and astrocytes under GluTox and oxygen-glucose deprivation (OGD) conditions. Reducing [Ca²⁺]i inhibits necrotic cell death in an acute experiment. Incubation of cerebral cortex cells with 3-EA leads to an overexpression of anti-apoptotic genes BCL-2, STAT3, SOCS3, along with downregulation of genes TRAIL, MLKL, Cas-1, Cas-3, IL-1β and TNFα. Intraperitoneal administration of 3-EA reduces the volume of necrotic areas, perinecrotic edema, cell damage, and neurological deficits in rats with MCA occlusion.

Conclusion: 3-EA dose-dependently suppresses the death of cerebral cortex cells under the excitotoxic effects of glutamate and ischemia/reoxygenation. Cell-protective effect of 3-EA involves changes in the basal and ischemia/reoxygenation-induced expression of genes encoding anti-apoptotic proteins and oxidative status proteins, which leads to inhibition of the late irreversible stages of apoptosis. A course administration of 3-hydroxypyridine ascorbate at a dose of 18 mg/kg per day reduces the severity of damage both by preserving the population of neurons in the penumbra zone and by limiting the local stress response.

Efficacy of various combined treatment regimens in patients with stable effort angina, functional classes II-III

Svetlana G. Dorofeeva, Eugenia N. Konoplya, Oksana V. Mansimova — Sat, 17 Aug 2024 00:00:00 +0300

Introduction: Inflammation and metabolic disorders of cardiomyocytes are undoubtedly important pathogenetic links in the development of coronary heart disease and its complications. Our study assessed the effect of various combined treatment regimens on cycle ergometry performance and plasma concentrations of pro- and anti-inflammatory interleukins in patients with stable effort angina, functional classes II-III.

Materials and Methods: The clinical study included 120 patients diagnosed with coronary heart disease: stable effort angina, functional classes II–III, and 40 healthy participants who met the inclusion criteria. Further, four groups were randomly formed: a group receiving the standard treatment; a group receiving Mexicor for 10 days in addition to the standard treatment; a group receiving Mexicor and Phosphogliv for 10 days in an addition to the standard treatment; and a group receiving Mexicor and Polyoxidonium for 10 days in addition to the standard treatment. After the treatment, cycle ergometry performance and plasma concentrations of pro- and anti-inflammatory cytokines were assessed in the patients. Mathematical statistical analysis was carried out using Statistica 10.0 software. The statistical significance of differences between the qualitative indicators was assessed using the χ2 test. The results of the statistical analysis were considered statistically significant at p<0.05.

Results: The combined treatment with using the above-mentioned pharmacological agents resulted in a statistically significant improvement in cycle ergometry indicators: Mexicor made it possible to increase the threshold load power by 43.0% and the total load power – by 70.3%. In the groups of patients with coronary heart disease who had received Mexicor and Phosphogliv or Mexicor and Polyoxidonium, there was an increase in the load duration by an average of 69.0% and in the rate-pressure product – by 25%, respectively. When analyzing the dynamics of plasma concentrations of pro-inflammatory cytokines, it was found that a statistically significant decrease in the concentrations of tumor necrosis factor-α, interleukin-1ß, interleukin-6 and interleukin-10 was registered only in the groups receiving Mexicor and Phosphogliv or Mexicorand Polyoxidonium in addition to the standard treatment.

Conclusion: Applying the combined treatment regimens using Mexicor and Phosphogliv or Polyoxidonium results in the most effective improvement of the cycle ergometry performance and the dynamics of plasma concentrations of pro- and anti-inflammatory interleukins in patients with stable effort angina, functional classes II-III.

Chemoproteomic analysis of the promising candidate molecule of the indole derivative with lab code SV-1010 and other non-steroidal anti-inflammatory drugs

Fri, 19 Jul 2024 00:00:00 +0300

Introduction: For effective and safe pharmacotherapy of pain, it is important to evaluate the mechanisms and spectrum of action of non-steroidal anti-inflammatory drugs (NSAIDs), including their effect on the proteom, central effect, as well as pain relieving and anti-inflammatory effects. The aim of the study was to evaluate the complex of differences between the promising candidate-molecule of indole derivative SV-1010 and the well-known NSAIDs.

Materials and Methods: Chemoproteomic moduling of pharmacological effects of SV-1010 and NSAID diclofenac, nimesulide and ketorolac on the rat proteom by means of topological analysis of chemographs.

Results: The significant differences in the effects of the studied molecules were found for 820 proteins of the rat proteom. SV-1010, to a lesser degree than the other molecules, can inhibit dopamine D1- and D2-type receptors and, at the same time, stimulate the release of dopamine in the neostriatum (EC50 = 27 nM). SV-1010, to a greater extent than the other molecules, can inhibit the GABA conveyor (EC50 = 65 nM) and the NMDA receptors Grin1/Grin2b (IC50 175 nM). SV-1010 can activate Cannabinoid CB2 receptors, inhibit enzymes of leukotriene biosynthesis, CC receptors of pro-inflammatory chemokines and leukotrienes.

Conclusion: The chemoreactomic and chemoproteomic profiling of SV-1010 indicated its potential central effect through dopaminergic and GABA-neurotransmission and additional anti-inflammatory mechanisms, which can help increase pain-relieving effects.

Ethical aspects of clinical trials in Russia and BRICS countries: an overview

Tue, 17 Sep 2024 00:00:00 +0300

Introduction: In the context of the globalization of the clinical trials market, the rapid growth in their number, the fast development of biomedical research using new technologies, and insufficient control over their conduct by state regulatory authorities and independent ethics committees, ethical aspects of conducting clinical trials and the issue of protecting patients’ rights remain relevant. The aim of the study: To review and compare the legislative frameworks and regulations of ethical aspects of clinical trials in Russia and the BRICS countries, which possess significant scientific, industrial, and economic potential – China, India, and Brazil.

Material and Methods: The search was conducted using PubMed, Medline, and Google Scholar databases, with descriptors including ethics in clinical trials, legislative regulation of clinical trials, and ethic committee. The selection criteria included publications from 2010 to 2024 and articles focusing on the regulation of clinical trials in Russia, China, India, and Brazil, along with their histories and evaluation forms.

Results and Discussion: In Russia, the work of Ethics Committees is based on the European model, grounded in modern international ethical norms and regulatory documents of the Russian Federation and the Eurasian Economic Union (EAEU). A drawback is the lack of structured interaction between Ethics Committees at both national and local levels. The work of Ethics Committees in China and India faces several problems, such as weak organizational structure, unjustified membership composition, low training, incompetence, weak control and management mechanisms, and flawed systems for obtaining informed consent. The Brazil’s ethical and regulatory system meets global requirements and ethical standards, aimed at protecting the rights of clinical trials participants. However, in all developing countries, there remains a potential danger for clinical trial participants with a low socio-economic standard of living.

Conclusion: The experience of BRICS countries, which are intensively developing in the field of clinical trials, is interesting in terms of developing possible approaches to monitoring activities, ensuring interaction, certifying Ethics Committees, and centrally training Ethic Committee members in Russia.

Study of the neuroprotective properties of the heteroreceptor EPOR/CD131 agonist of peptide structure in tau-proteinopathy modeling

Mon, 17 Jun 2024 00:00:00 +0300

Introduction: Tau proteinopathy is a pathology associated with the activation of post-translational modifications and interactions of pathophysiological cascades of neuroinflammation with hyperphosphorylation of Tau aggregates. Therefore, preference is given to agents that have properties in reducing or slowing down the processes of neuroinflammation and post-translational modifications in the brain.

Materials and Methods: The study was conducted on male and female homozygous individuals of a transgenic murine line with overexpression of mutant human Tau gene (P301S) and a background wild mouse line C57Bl/6J. To assess the progression of Tau proteinopathy, behavioral tests were used at two control time points, and the last one measured the level of neuroinflammation markers and tau-proteinopathy.

Results and Discussion: In the group of P301S mice treated with ARA-290, an improvement in the phenotypic picture of Tau proteinopathy was demonstrated compared with intact animals. In the Barnes circular maze test, mice showed a decrease in the total distance traveled and the latent time spent on the platform, which indicates a rapid entry into the shelter. In the O-shaped maze test, the group maintained a fairly high level of spontaneous exploratory behavior. In the vertical rod test, the animals recorded the best time indicators that they needed to turn and maintain balance compared to the intact group. A statistically significant decrease in the level of GSK-3β and an increase in CDK5 and PP2A were revealed, which indicates a dephosphorylating effect on Tau protein, as well as markers of neuroinflammation. NF-KB and TNF-α were significantly reduced by 57% and 32%, respectively, compared to the intact group.

Conclusion: In the model of transgenic P301S murine line with overexpression of the mutant human Tau gene, the peptide agonist of the EPOR/CD 131 heteroreceptor demonstrated neuroprotective properties, which were confirmed by indicators of behavioral tests and markers of neuroinflammation and tau-proteinopathy.

Research on the influence of Siberian fir polyprenols on learning and memory of mice with an experimental model of Alzheimer's disease

Nikolay I. Suslov, Yulia S. Fedorova, Maxim L. Korobov — Thu, 08 Aug 2024 00:00:00 +0300

Introduction: Alzheimer’s disease is increasingly becoming a cause of early disability and death. Attempts to create a pharmacotherapeutic agent that provides an effective result in treating this pathology have so far been unsuccessful. This article presents the results of experimental studies aimed at creating a more effective treatment of Alzheimer’s disease.

Materials and Methods: The object of the study was polyisoprenoids isolated from Siberian fir (Abies sibirica Ledeb.). Experiments were conducted on 108 outbred adult male mice from the CD1 (cluster of differentiation 1) stock, weighing 28-30 g (at 5 weeks old). The effect of the sum of polyisoprenoids on learning and memory was studied in a dose range of 5, 20, 50, 100, 200, and 500 mg/kg. Gliatilin was used as a reference drug at a dose of 90 mg/kg. The experimental model of cognitive dysfunction was created by chronic (for 20 days) intraperitoneal injection of scopolamine at a dose of 2 mg/kg. Cognitive dysfunction was assessed by changes in the indicators of conditioned passive avoidance reflex (CPAR).

Results and Discussion: The results of the experiments showed that chronic administration of scopolamine caused a pronounced decrease in the reproducibility of CPAR. Simultaneously, the studied substance at doses of 20 and 50 mg/kg restored the reproducibility of the reflex to a level close to the values of intact control. Gliatilin showed a similar effect. The authors believe that cholinergic receptors are involved in the development of the anti-amnestic effect.

Conclusion: The presented study demonstrates the discovery of a new agent with anti-amnestic action based on the sum of polyprenols from Siberian Fir (Abies sibirica Ledeb.), family Pinaceae. This agent can be used for the treatment of neurodegenerative pathologies, particularly Alzheimer’s disease.

Development of a transcutaneous form of desloratadine

Fri, 28 Jun 2024 00:00:00 +0300

Introduction: For people with allergies, it is not always possible to prevent allergens admission to the body. For this reason, it is essential to create a long-acting drug that can prevent an allergic reaction. The very promising option is a transdermal therapeutic system (TTS), with H1 blockers. The aim of the study: To create a technology for the production of desloratadine-based TTS, suitable for routine use with prolonged drug releasing.

Materials and Methods: Polyvinylpyrrolidone K30 (PVP) was used as the main matrix polymer for TTS, and the Kruofilm polymer membrane was used as the adhesive layer. The formation of the polymer membrane was carried out by drying a water-alcohol solution of the components at a temperature of 60ºC. Characterization of the obtained dosage forms was made by spectrophotometry. The resulting micelles size was measured by dynamic light scattering.

Results and Discussion: Round shaped TTS (patches) with water-dissolved DL and its micellar form were obtained. The size of micelles with DL was 22.8±5.2 nm. The spectrophotometric method for determining DL has been developed. The containing of DL in the patches ranged from 5 to 25 mg per patch, depending on the manufacturing technology.

Conclusion: We have developed TTSs containing DL by a solution and a micellar form. A top adhesive layer has been selected that has moisture resistance, strength and reliable fixation of the matrix with the active substance on the skin. A method for the spectrophotometric determination of DL in solutions and patches was developed and validated.

Synthesis and pharmacological properties of novel guanidine derivatives of quinazoline-2,4(1H,3H)-dione

Mon, 23 Sep 2024 00:00:00 +0300

Introduction: Na⁺/H⁺ exchanger type 1 (NHE-1) is a validated drug target for the treatment of cardiovascular and ophthalmic diseases due to the cytoprotective, anti-ischemic and anti-inflammatory properties of NHE-1 inhibitors. This article presents data on the synthesis and pharmacological activity studies of novel guanidine derivatives of quinazoline-2,4(1H,3H)-dione 6-11 and reference drugs amiloride, rimeporide, zoniporide, dexamethasone, aminoguanidine, and acetylsalicylic acid.

Materials and Methods: Pharmacological properties were assessed using pH-dependent platelets deformation assay, anti-inflammatory activity assay on LPS-stimulated peritoneal macrophages, antiglycation assay, analysis of platelet aggregation in vitro and measurement of intraocular pressure in vivo.

Results: Several compounds combine NHE-1 inhibition with antiglaucomic and antiplatelet activity. Compound 11 significantly inhibits pro-inflammatory activation of murine macrophages (IC₅₀15.64 μM) and effectively suppresses the formation of glycated proteins (38.1±2.6% in C 1 mM).

Conclusion: The investigated compounds represent a promising scaffold for development of agents for the treatment of cardiovascular pathologies, glaucoma, excessive inflammation, and late diabetic complications including retina diabetics and thrombosis.

New compounds with a dihydropyridine framework as promising hypolipidemic and hepatoprotective agents

Elena S. Ketova, Anna V. Myazina, Elena Yu. Bibik, Sergey G. Krivokolysko — Mon, 23 Sep 2024 00:00:00 +0300

Introduction: To solve the problem of complex and safe pharmacological correction of metabolic disorders, including hyperlipidemia, hyperglycemia, and liver lesions is currently very important. With this in mind, the new derivatives of cyanothioacetamide with a dihydropyridine framework, with a potential effect on lipid and carbohydrate metabolism and the functioning of the liver are of great interest.

Materials and Methods: To conduct an experiment, three samples were selected from an extensive library of new cyanothioacetamide derivatives, which, according to in silico studies, proved promising towards the positive effect on lipid and carbohydrate metabolism, as well as towards the protective effect on the liver. To study the compounds in vivo, metabolic disorders were simulated in Wistar rats by long -term alimentary and subsequent dexamethasone loads. The pharmacological efficacy of new compounds AZ-383, AZ-257, AZ-020 (administered in a dose of 1 mg/kg for 14 days) was assessed in comparison with metformin (300 mg/kg for 14 days) and vildagliptin (8 mg/kg for 14 days) by determining the biochemical indicators of blood (ALT, AST, total bilirubin, total cholesterol, triglycerides, and glucose), morphological and morphometric studies of the liver sections, and the study of immunohistochemical indicators of hepatocyte proliferation. The experimental results were statistically processed using the recognized methods of mathematical statistics. When processing the experimental data, the average arithmetic (AA) was determined. The statistical significance of the compared options was determined on the basis of the Student’s t-test, with the critical value of the Student’s t-test equal to 2.101 and the significance level α = 0.05.

Results: The study shows that all the new compounds studied in the experiment – AZ-383, AZ-257, AZ-020 – have hypolipidemic and hepatoprotective properties, which manifested in the reduced levels of liver biochemical markers of blood, which had increased after simulating metabolic disorders, in the reduced concentration of total cholesterol and triglycerides in blood, in the normalization of liver microarchitecture, as well as in the proliferative activity of hepatocytes.

Discussion: The hepatoprotective and hypolypidemic properties of the new derivatives of cyanothioacetamide with a dihydropyridine framework, which were determined while conducting the experiment, can be accounted for by their effects on the biotargets, identified for AZ-383, AZ-257, and AZ-020 in silico. According to the results, the most pronounced hepatoprotective activity was found in AZ-383 (intragastrically, 1 mg/kg for 14 days). The Ki-67 proliferation index under the influence of this compound was registered at the level of 1.48±0.03%, which exceeds this indicator in the control animals and proves a significant hepatoprotective activity of AZ-383.

Conclusion: The results show good prospects and high efficacy of the new studied cyanothioacetamide derivatives, such as AZ-383, AZ-257, AZ-020 (in a dose of 1 mg/kg), in terms of comprehensive correction of metabolic disorders. Further study is needed for this class of compounds.

Identification and synthesis of metabolites of the new 4.5-dihydroisoxazol-5-carboxamide derivate

Sun, 30 Jun 2024 00:00:00 +0300

Introduction: 3-(2-butyl-5-chloro-1H-imidazole-4-yl)-N-[4-methoxy-3-(trifluoromethyl)phenyl]-4,5-dihydro-1,2-oxazole-5-carboxamide is new antirheumatic drug. It is necessary to identify and synthesize the biotransformation products for its complete pharmacokinetic study.

Materials and Methods: A biotransformation study was carried out by intraperitoneal administration of the drug to Wistar rats and Soviet Chinchilla breed rabbits. Animal blood sampling was performed before the injection and 0.5 h, 1 h, 2 h, 4 h, 24 h after the injection of the investigated compound. The samples were immediately centrifuged for plasma separation. Urine was simultaneously collected from rats before the administration and at intervals of 0-2 h, 2-4 h, 4-6 h, 6-24 h after administration, faeces – before administration and at intervals of 0-12 h and 12-24 h after administration. The samples were analyzed by HPLC-MS/MS after immediate preparation by adding acetonitrile.

Results and Discussion: 3-(2-butyl-5-chloro-1H-imidazole-4-yl)-4,5-dihydro-1,2-oxazole-5-carboxylic acid and 4-methoxy-3-(trifluoromethyl)aniline – hydrolysis products of the active substance were found during the analysis of plasma, urine and fecal samples. The 4,5-dihydro-1,2-oxazole-5-carboxylic acid derivative has been synthesized. The second metabolite is a raw material for production of active pharmaceutical substance. During comparative tests, no significant difference between the retention times, ratio areas of chromatographic peaks at the main MRM-transitions and mass spectra of these metabolites on chromatograms of standard and animal samples was found, which indicates the correct identification of biotransformation products.

Conclusion: The studied drug undergoes biotransformation by hydrolysis to form two main metabolites: 3-(2-butyl-5-chloro-1H-imidazole-4-yl)-4,5-dihydro-1,2-oxazole-5-carboxylic acid and 4-methoxy-3-(trifluoromethyl)aniline. The structure of the metabolites was confirmed by comparison with the synthesized standard samples using HPLC-MS/MS.