Grigory V. Mokrov

Introduction: Exchange proteins directly activated by cyclic AMP (Epac) represent a promising therapeutic target for developing novel cardiotropic agents. Epac-dependent signaling regulates intracellular calcium dynamics, intercellular coupling, and the contractile function of cardiomyocytes. Preclinical studies have demonstrated the potential of Epac2 inhibitors for the treatment of cardiovascular diseases (CVDs). Methods: New potential Epac protein inhibitors from the series of arylsulfonylaminopyridines were designed using pharmacophore modeling, molecular docking, ADMET analysis, and molecular dynamics. The compounds were synthesized via the reaction of N-substituted aminopyridines with arylsulfonyl chlorides. The potential cardiotropic activity of the synthesized compounds was evaluated ex vivo using a model of an isolated rat atrial myocardial strip. results: Pharmacophore-based design led to novel aminopyridine derivatives predicted to interact with Epac2. Computational studies suggested stabilization of the inactive conformation of Epac2. Ex vivo experiments demonstrated modulation of myocardial contractility consistent with a potential Epac-related mechanism. Results: Pharmacophore-based design led to novel aminopyridine derivatives predicted to interact with Epac2. Computational studies suggested stabilization of the inactive conformation of Epac2. Ex vivo experiments revealed changes in myocardial contractility that are consistent with the proposed hypothesis of Epac2 involvement, although further studies are required to establish direct target engagement. Discussion: A key objective of this study was to refine the ligand-binding pocket of Epac2, as preliminary in silico analyses revealed discrepancies with previously reported models. In parallel, the potential for PKA-independent Epac2 involvement was assessed using ZMEI-26 in a forskolin-based experimental setup. The combined computational and functional data suggest possible Epac2 engagement and are consistent with the proposed mechanism, although further studies are required to confirm direct target interaction. Conclusion: A new series of aminopyridine derivatives was created using molecular modeling methods as potential Epac2 inhibitors. During the synthesis, 11 compounds (coded ZMEI) were obtained, several of which (ZMEI-3, ZMEI-15, ZMEI-18, ZMEI-19, ZMEI-22, ZMEI-26, and ZMEI-14) demonstrated the ability to reduce the automaticity and contractility of isolated rat myocardial strips.

Background. Parkinson’s disease has a multifactorial pathogenesis involving glutamate excitotoxicity, oxidative stress, neuroinflammation, and dysregulation of neurotransmitter systems. Therefore, the development of multitarget compounds capable of interacting with several pathogenetically relevant targets is a promising research direction. Objective. Evaluation of the interaction of the hybrid molecule ADK-1113, designed on the basis of structural fragments of fabomotizole and memantine, with σ₁, NMDA, and AMPA receptors using molecular modeling methods. Methods. Molecular docking of ADK-1113 was performed using the structures of σ₁-, NMDA-, and AMPA-receptors, followed by molecular dynamics (MD) simulations of the resulting complexes. Binding stability was assessed based on ligand retention within the binding site, root mean square deviation (RMSD) values, the pattern of noncovalent interactions, and binding free energy calculated using the MM/PBSA method (Molecular Mechanics/Poisson–Boltzmann Surface Area). Results. ADK-1113 bound to all studied receptors and remained within their binding sites during molecular dynamics simulations. The most stable complex was observed for the σ₁-receptor, which was consistent with the most negative binding free energy values obtained by MM/PBSA. The complex with the NMDA-receptor showed intermediate stability, whereas the least favorable binding energy values were obtained for the AMPA-receptor. Conclusion. The obtained data indicate the potential multitarget activity of ADK-1113 and its ability to interact with several components of neurotransmitter systems. The results of in silico modeling require further experimental confirmation.

Introduction. Matrix metalloproteinases (MMPs) belong to the family of zinc-dependent endopeptidases capable of degrading the main components of extracellular matrix proteins that form the basis of connective tissue; they are also involved in many pathological processes in the body. MMP-2 is actively expressed in lung cancer tissues, while MMP-2 and MMP-9 play a significant role in the development of colorectal cancer. Objective. To evaluate the effect of 1-({4-[(2,4-dichlorobenzoyl)amino]phenyl}sulfonyl-(2S,4R)-4-hydroxypyrrolidine-2 carboxylic acid (GGM-27) on tumor growth and survival in BALB/c mice using the AKATOL colon adenocarcinoma model. Materials and methods. AKATOL colon adenocarcinoma was implanted subcutaneously into the axillary region of male BALB/c mice at a dose of 1×10⁶ cells. From day 2 to day 15 of tumor development, GGM-27 was administered intraperitoneally (i.p.) at doses of 1 mg/kg and 10 mg/kg. Doxorubicin was used as a reference drug, administered at a dose of 5 mg/kg i.p. on days 2 and 4 of tumor development. The antitumor activity of GGM-27 was assessed by tumor growth inhibition (TGI) and lifespan extension (LSE). Results. After a 14-day course of i.p. administration of GGM-27 at a dose of 1 mg/kg, TGI was 72.8 % on day 21 of the experiment and 71 % on day 28. After i.p. administration of GGM-27 at a dose of 10 mg/kg, TGI was 63 % on day 21 and 56% on day 28. LSE after a course of GGM-27 at a dose of 10 mg/kg was 24 %. After two i.p. administrations of doxorubicin at a dose of 5 mg/kg, TGI was 82 % on day 21 and 69 % on day 28; however, two of the 12 animals died due to drug toxicity. Conclusions. Administration of 1-({4-[(2,4-dichlorobenzoyl)amino]phenyl}sulfonyl-(2S,4R)-4-hydroxypyrrolidine-2-carboxylic acid at a dose of 1 mg/kg caused TGI. On day 21 of the experiment, with GGM-27 administration, TGI was 73 %, and on day 28, TGI was 71 %.

Introduction. As a screening result in a number of alkoxyphenyltriazoalkanes, the compound ALM-802, which in the form of its trihydrochloride has pronounced antiarrhythmic and antiischemic activity, was identified at the Federal State Budgetary Scientific Institution «Federal Research Center for Innovator and Emerging Biomedical and Pharmaceutical Technologies». However, the pH of the aqueous solution of this salt is quite low – 3.5, – which does not exclude the possibility of some local irritant activity in injectable dosage forms of the compound. Therefore, the succinate and citrate of compound ALM-802 – ALM-802S and ALM-802C, respectively, – were synthesized. The pH of the aqueous solutions of these compounds is significantly closer to neutral (ALM-802S – 5.8 and ALM-802C – 5.55), which does not suggest their local irritant activity, suggesting no local irritancy. Objective. To study in a comparative aspect acute toxicity with a 24-hour observation period, antiarrhythmic and anti-ischemic activity of compounds ALM-802T, ALM-802S and ALM-802S. Materials and methods. The toxicity parameters were determined on white outbred male mice, the calculation was carried out according to Finney. The antiarrhythmic activity of the compounds (2 mg/kg) was studied on a model of aconitine cardiac arrhythmias in male white rats. The anti-ischemic effect of the substances was tested on a model of acute subendocardial ischemia caused by the nonselective beta-adrenomimetic isoproterenol (20 mg/kg/min for 5 minutes). The intensity of ischemic damage was judged by the magnitude of ST segment depression on the ECG (standard lead II) 5 minutes after the start of the isoproterenol infusion. Results. Analysis of the data obtained showed that the LD50/24 of the studied ALM-802 salts are comparable, they belong to the IV class of toxicity – low-toxic compounds. They are equally highly effective in models of aconitine heart rhythm disorders and subendocardial ischemia. Conclusion. The ALM-802T, ALM-802S and ALM-802S compounds on the studied models do not differ in efficiency.

The synthesis, purification, and comprehensive characterization of the E- and Z-isomers of the prospective anticonvulsant compound GIZH-298 were carried out. It was demonstrated that both isomers could be obtained in a pure form by isolating the corresponding oximes at the initial stage of synthesis. The E- and Z-isomers exhibit pronounced differences in their physicochemical characteristics, spectral features, and pharmacological profiles. Z-GIZH-298 was found to be photosensitive, undergoing Z-E isomerization upon irradiation, as confirmed by both theoretical analysis and experimental observations. Additionally, partial conversion to the E-form occurs during prolonged reflux of Z-GIZH-298 in polar solvents, whereas the compound remains stable in aqueous and acidic media at ambient temperature. In the maximal electroshock assay, both isomers demonstrated anticonvulsant activity, with Z-GIZH-298 showing substantially higher efficacy than its E-counterpart. These findings highlight the need to account for potential E-isomer formation when considering further development of Z-GIZH-298 as a promising anticonvulsant agent.

Objective. To study the potential use of Epac regulatory protein inhibitors to prevent alcohol-induced myocardial fibrosis. Materials and Methods . To simulate holiday heart syndrome (HHS), rats were given a 10 % aqueous ethanol solution as the sole fluid source for 10 days, followed by drinking water for 10 days, and then a 10 % aqueous ethanol solution for the following 10 days. Connective tissue and adipose tissue density were assessed in histological preparations obtained from atrial, interatrial septal, and ventricular tissues stained with gallocyanin-eosin and picrofuchsin according to Van Gieson. A scoring system was used to quantify the intensity of the identified pathological changes. Results. Following experimental therapy with ZMEI-3, the degree of connective tissue proliferation in the right atrium and interatrial septum in these animals was significantly lower than that observed in the control group (p = 0.0079 and p = 0.0013, respectively). The intensity of fatty deposits was significantly lower in the right and left atria (p = 0.035 and p = 0.0034, respectively). Conclusion . Experimental therapy with ZMEI-3 in animals with HHS syndrome significantly reduced both the intensity of connective tissue and adipocyte deposits in atrial myocardium and interatrial septum, which may underlie the antiarrhythmic effect of the compound.

Introduction. The compound ALM-802S, which has a pronounced cardiotropic effect, has been identified among the alkoxyphenyltriazoalkanes. The structure of this substance includes 2 pharmacophore components of the cardioprotective agent trimetazidine, which can potentially cause extrapyramidal disorders. The ALM-802S compound, in addition to its cardiotropic activity, has anxiolytic and analgesic effects. It is possible that the ALM-802S compound, acting on the central and peripheral serotonergic and dopaminergic mediator systems, may, like trimetazidine, cause central side effects. The study aim. It was to evaluate the effect of the ALM-802S compound on the content of biogenic catecholamines, indolamines and their metabolites in blood plasma and separate brain structures in rats. Materials and methods. The experiments were carried out on white male rats weighing 180–200 g. The test substance was administered at a dose of 5 mg/ kg i./p. 60 minutes before decapitation. The content of indolamines and catecholamines in blood plasma and brain structures was determined using HPLC/ED. Results. Analysis of the obtained data showed that the compound ALM-802S with a single systemic administration does not affect the content of indolamines in blood plasma and brain structures of rats. Measurements of the catecholamine content showed that the ALM-802S compound significantly reduced the concentration of DOPA and HCV in blood plasma, the content of A, HA, DA, DOPA and the ratio of metabolites remained virtually unchanged. Of all the studied brain structures, only in the hypocampus there was a decrease in the GVK/DA ratio, which characterizes the DA decay rate. No changes were found in the rest of the brain structure. Conclusion. Using neurochemical analysis, information was obtained on the absence of a pronounced effect of ALM-802S on the functional activity of the central dopaminergic and serotonergic systems, which minimizes the possibility of extrapyramidal disorders and the development of drug dependence when using ALM-802S in therapeutic doses.

Background. Matrix metalloproteinases (MMPs) are a group of zinc dependent enzymes that play a key role in regulating both physiological and pathological processes in humans, particularly in oncological diseases. MMP-2 (gelatinase A) overexpression promotes metastatic progression in tumor cells. The development of selective MMP inhibitors as anticancer agents represents a promising approach in modern pharmacology. Previously, the compound 1-({4-[(4-chlorobenzoyl)amino]phenyl}sulfonyl)-L-proline, which received the laboratory code AL-828, was designed and synthesized using the pharmacophoric approach, possessing inhibitory activity against MMP-2.Objective. Antitumor and antimetastatic activity of AL-828 compared with gemcitabine as the first-line antitumor drug.Methods. This study was conducted on an experimental model of Lewis lung carcinoma (LLC) in C57BL/6 mice. AL-828 was administered intraperitoneally (i.p.) at doses of 10 and 30 mg/kg from day 1 to day 14 of tumor development. The antitumor drug gemcitabine was administered intraperitoneally as a reference drug at a dose of 50 mg/kg on days 2 and 9 of tumor development. The antitumor effect was assessed on days 7, 9, 15, and 21 of tumor development, and the metastasis inhibition index (MII) was measured on day 21 of tumor development. In this study, the average lifespan and survival rates were measured. Survival analysis was performed using the Kaplan–Meier method.Results. AL-828 at doses of 10 and 30 mg/kg did not cause inhibition of tumor growth (ITG), whereas gemcitabine at a dose of 50 mg/kg significantly reduced tumor growth at all stages of registration at the level of 60 %. AL- 828 at a doses of 10 and 30 mg/kg, AL-828 showed antimetastatic effects, with MII values of 48.3 % and 39.2 %, respectively. Gemcitabine showed an MII of 75.8 %. Lungs from 50 % of the control group exhibited both light and high colonization potential with metastases. In contrast, the administration of either gemcitabine or AL-828 resulted in only light colonization potential in 90 % of treated animals.Conclusion. Systemic course administration of AL-828 demonstrated antimetastatic activity in a model of LLC.