Research Results in Pharmacology

Efficacy and side effects of different regimens of experimental postmenopausal osteoporosis therapy with zoledronic acid and their correction by combination with resveratrol

Olga A. Shevchenko, Alexander A. Dolzhikov, Oleg S. Gudyrev — Fri, 26 Jun 2026 00:00:00 +0300

Introduction: As an age-associated metabolic disease of the skeleton, osteoporosis (OP) causes a significant medical and socio-economic burden. One of the challenges in clinical practice is the adverse effects of widely used bisphosphonates. Therefore, the search for preventive pharmacotherapeutic strategies is needed.

Materials and Methods: Sixty female Wistar rats randomized into 6 equal groups were used to study the effects of a single and prolonged intravenous administration of zoledronic acid (ZA) without and in combination with an endothelioprotective dose of resveratrol (2 mg/kg intraperitoneally daily), compared with sham-operated and osteoporotic animals. X-ray densitometry and histological methods were used.

Results and Discussion: Already at 4 weeks after bilateral ovariectomy (OVx), the typical signs of osteoporotic lesions comparable with OP in humans are present. A single administration of ZA following 4 weeks of OP had a significant corrective effect on bone structure and the fatty component of bone marrow. After a second dose of ZA at 8 weeks after OVx, signs of bone damage were detected by week 12. The addition of 4 weeks of resveratrol therapy after the second dose of ZA (until 12 weeks after OVx) leads to switching of bone marrow precursors to osteoblastic differentiation, reducing the adipocytic component; cytoprotective action on osteocytes with their switching to a synthetic phenotype, reducing the potential activation of osteoclasts and blocking periosteocytic osteolysis; and protection of bone vessels via the endothelioprotective properties of resveratrol.

Conclusion: An endothelioprotective dose of resveratrol exerts multitarget cell- and endothelium-dependent mechanisms of protective action when combined with prolonged zoledronic acid therapy, preventing its adverse effects.

Evaluation of cardiotoxicity of a new antitumor compound, anthrafuran, a derivative of anthracycline antibiotics

Fri, 26 Jun 2026 00:00:00 +0300

Introduction: Anthracyclines and anthracenedione derivatives are a group of anticancer drugs that are widely used in clinical practice. Their application is frequently limited by cardiotoxic effects. The aim of this study was to evaluate the cardiotoxicity of the new anthracenedione derivative Anthrafuran (AF) in comparison with Doxorubicin (Dox) and Mitoxantrone (MT).

Materials and Methods: The study was conducted on 50 adult female Wistar rats. AF, MT, and Dox were administered in therapeutic doses and regimens for this animal species (AF: 20 mg/kg x 3/48 per os; MT: 1 mg/kg x 3/96 i.v.; Dox: 2.5 mg/kg x 3/72 i.v.). Electrocardiographic (ECG) parameters and heart weight index were determined; histological evaluation of the myocardium was performed on days 1 and 30 post-treatment.

Results: Dox intravenous injections in therapeutic doses caused a pronounced cardiotoxic effect. The signs of cardiac decompensation manifested as a significant decrease in heart rate (HR) (Dox 486±6, control 522±12.8), prolongation of the QT interval (Dox 0.06±0.002, control 0.048±0.001), an increase in weight index of the heart (Dox 0.41±0.03, control 0.36±0.006), and myocardial damage. Although the cardiotoxicity of MT was less pronounced, it nevertheless negatively affected both ECG parameters (HR 465±15, QT 0.056±0.002) and the structure of the heart muscle. The use of AF did not cause any pathological changes and had no effect on the heart weight index or ECG parameters (HR 498±7.3, QT 0.052±0.003, weight index of the heart 0.35±0.009).

Conclusion: Unlike Dox (2.5 mg/kg x 3/72 i.v.) and MT (1 mg/kg x 3/96 i.v.), the new anthracenedione derivative AF (20 mg/kg x 3/48 per os) in therapeutic doses does not possess cardiotoxic properties, supporting its potential for clinical study.

Synthesis and ophthalmic hypotensive effect of new potential benzimidazole-based Rho-kinase-2 inhibitors

Fri, 26 Jun 2026 00:00:00 +0300

Introduction: Based on the chemical structures of known Rho-kinase-2 inhibitors, 18 benzimidazole derivatives were synthesized and studied for their ophthalmic hypotensive activity in animals with normal intraocular pressure. The dependence of the pharmacological effect on the chemical structure of the compounds was analyzed. The effect of the most active compounds on Rho-kinase-2 activity was assessed.

Materials and Methods: Ophthalmic hypotensive activity was assessed by measuring intraocular pressure with a TonoVet veterinary tonometer in 120 mongrel rats (6 in each group) before and after instillation of reference drug solutions (timolol and melatonin) and 18 test substances. The effect of the test compounds on Rho-kinase activity was assessed using an in vitro enzyme-linked immunosorbent assay spectrophotometrically.

Results: The most active compounds among the new benzimidazole derivatives after a single instillation at a concentration of 0.4% were compound 7a (1-(4-fluorobenzyl)-3-(2-(pyrrolidin-1-yl)ethyl)-1,3-dihydro-2H-benzo[d]imidazol-2-imine hydrochloride), which reduced intraocular pressure in normotensive animals by 28.21%, exceeding the effect of the reference drug timolol(26.84%), but inferior to melatonin (30.95%), and compound 1d (1-(1-(2-(azepan-1-yl)ethyl)-1H-benzo[d]imidazol-2-yl)-3-(3-trifluoromethyl)phenyl)urea hydrochloride), which reduced ophthalmotonus by 23.96%, slightly inferior to timolol. The test compounds do not affect intraocular pressure dynamics in the contralateral eye and, therefore, do not have a systemic effect, unlike the reference drugs. It was also found that compounds 7a and 1d at a concentration of 1*10-4 mol/L inhibit Rho-kinase-2 by 26.72% and 18.11%, respectively.

Conclusion: The most active compounds, 7a and 1d, were identified as Rho kinase-2 inhibitors that exhibit ocular hypotensive effects in normotensive animals in vivo.

From PDC to integral quantitative metrics: an innovative approach to assessing adherence to combination pharmacotherapy in outpatient practice

Sergey B. Fitilev, Irina I. Shkrebniova, Dmitry A. Klyuev — Fri, 29 May 2026 00:00:00 +0300

Introduction: Current methods for assessing adherence to combination therapy based on aggregated measures (e.g., PDC, proportion of days covered) poorly capture treatment intensity and sustainability over time. We attempted to describe adherence trajectories for combination therapy using integral metrics. Aim: To develop a set of integral metrics analogous to key pharmacokinetic (PK) parameters – area under the concentration-time curve (AUC, area under the curve), maximum concentration (C_max), and time to maximum concentration (T_max) – to assess adherence to combination therapy as time-based exposure on the PDC methodology among outpatients.

Materials and Methods: As a specific example and a substrate for method development, we used results from a retrospective pharmacoepidemiologic cohort study of patients with heart failure (HF) who had experienced an acute myocardial infarction. Data were extracted from the Unified Medical Information and Analytical System of Moscow (Russia). Three patients with a follow-up duration of ≥24 months were randomly selected. We analyzed benefit-covered electronic prescription fills for key HF therapy classes and calculated adherence for each class using PDC across four half-year intervals. We constructed a stepwise trajectory of combination therapy as the function N(t), where N is the number of therapy classes for which the patient was covered at a given time point based on dispensed supply (accounting for days’ supply and overlaps), and t is follow-up time. By analogy with the “concentration-time” curve, we considered N(t) as a dynamic “therapy class coverage curve” and used it to calculate PK-analog adherence metrics: exposure to actually dispensed combination therapy, AUC_N(t)(class-months); normalized AUC_norm(0-1); C_maxN(t) (number of classes); and T_maxN(t) (months). In addition, we set a target threshold of N(t)≥3, determined the time to first attainment, T_optN(t), and calculated the time (or proportion of time) with N(t)≥k, denoted as T_N(t)≥k. We also quantified therapy “losses” between intervals based on a decrease in the modal value of N(t) and failure to reach the threshold over the follow-up period.

Results: AUC_N(t) values were 26.0, 87.37, and 36.67 class-months, and AUC_norm values were 0.27, 0.74, and 0.39 in patients with internal IDs 13, 39, and 110, respectively. The proportion of time with N(t)≥3 was 0.079, 0.96, and 0.23 (1.9, 22.9, and 5.5 months), and the modal N(t) values by half-year corresponded to patterns of 2→0, 3→3→4→4, and 3→2→1→0. The numeric profiles were consistent with the stepwise N(t) trajectories and reflected three distinct adherence patterns.

Conclusion: The proposed PK-associated approach enabled quantification of complex adherence trajectories for combination therapy in HF by representing them as an interpretable set of integral numeric metrics. These metrics characterize time-based exposure to combination therapy while accounting for adherence. The results support the conceptual validity of the method and may serve as a basis for further studies of its prognostic value.

Effect of solid herbal extract of Primula veris L. on the psychoemotional state of rats after chronic alcohol intoxication

Mon, 22 Jun 2026 00:00:00 +0300

Introduction: The aim of the study was the effect of solid herbal extract of Primula veris L. and comparison drugs phenotropil and mildronate on the psycho-emotional state of rats after chronic alcohol intoxication (CAI).

Materials and Methods: CAI was modeled by replacing drinking water with a 10% solution of ethyl alcohol with sucrose (50 g/L) for 6 months. After alcoholization, rats at the age of 16 months were divided into the following groups: 1 – intact – animals without CAI received oral 0.9% sodium chloride solution for 14 days; 2 – control – rats after ethanol withdrawal, which were injected with a 0.9% sodium chloride solution in a similar regimen; 3, 4 and 5 – experimental – females after CAI received oral solid herbal extract of Primula veris L. at a dose of 30 mg/kg, phenotropil 25 mg/kg, mildronate 50 mg/kg, respectively. After the end of treatment, to assess the psycho-emotional state of the animals, the Open field test, Elevated plus maze test, Marble burying test and Porsolt forced swim test were performed.

Results and Discussion: In animals after CAI, increased anxiety is observed, which manifested itself in a greater number of stands with support, urinations and boluses in the Open field test compared to the intact group. In the Elevated plus maze test, the trend remained the same: rats in the control group entered the open compartment installation less often, hung from it less often, and spent less time in the open compartment. In addition, females after CAI exhibited compulsive behavior in the Marble burying test. In the Porsolt forced swim test, animals in the control group had a shorter period of active swimming compared to intact females and a longer period of immobilization, which indicates depressive behavior in the former. Solid herbal extract of Primula veris L., phenotropil and mildronate contributed to the improvement of the psycho-emotional state of animals after CAI.

Conclusion: Thus, solid herbal extract of Primula veris L. at a dose of 30 mg/kg, which was administered orally once a day to female rats for 14 days after the withdrawal of ethanol, has a pronounced anxiolytic and antidepressant effect, while phenotropil and mildronate (25 mg/kg and 50 mg/kg, respectively, in a similar regimen) had a greater anticompulsive effect.

In silico exploring the mechanisms of action of diterpenoids from Rabdosia serra against lung cancer through inhibition of the anti-apoptotic pathway

Hung Duc Nguyen — Thu, 11 Jun 2026 00:00:00 +0300

Introduction: Apoptosis resistance in non-small cell lung cancer is frequently sustained by pro-survival Bcl-2 family proteins such as Mcl-1, motivating the search for new Mcl-1 inhibitors from natural products, including diterpenoids from Rabdosia serra.

Materials and Methods: An integrated in silico approach was applied to evaluate R. serra diterpenoids as putative Mcl-1 (PDB: 6QFQ) inhibitors, integrating molecular docking, molecular dynamics simulation, MM/GBSA rescoring, pkCSM-based ADMET prediction, and DFT calculations at the B3LYP/6-31G(d,p) level, with Tivantinib as the reference ligand.

Results and Discussion: Docking prioritized CPD1 (-10.31 kcal/mol) over Tivantinib (-9.09 kcal/mol). Molecular dynamics simulation indicated stable complexes, with CPD1-6QFQ showing a more compact, less solvent-exposed ensemble (Rg 1.42-1.45 nm; SASA 83-89 nm²; RMSD 0.09-0.15 nm). MM/GBSA favored CPD1 (ΔTOTAL -26.17 ± 2.79 kcal/mol) versus Tivantinib (-24.25 ± 4.50 kcal/mol). ADMET predicted high intestinal absorption for CPD1 (98.243%), a higher unbound fraction, and fewer liabilities (negative hepatotoxicity; negative hERG II inhibition). DFT supported CPD1 with a smaller ΔE (3.7165 eV) and higher softness (0.5381 eV⁻¹) than Tivantinib.

Conclusion: Convergent computational evidence nominates CPD1 as a leading putative Mcl-1 inhibitor for optimization, while target engagement, efficacy, and safety require experimental validation.

Morphometric visualization, analysis, and assessment of atherosclerotic lesions in an apolipoprotein E–deficient mouse model (ApoE−/−)

Petr R. Lebedev, Natalya V. Syrykh, Mikhail V. Pokrovskii — Thu, 19 Mar 2026 00:00:00 +0300

Introduction: The improvement of diagnostic methods in atherosclerosis is aimed at accurately determining the stage of the pathological process and eliminating subjective interpretation of data. Implementation of this approach increases the reliability of comparative assessment of the effectiveness of therapeutic and interventional strategies and reduces statistical variability within experimental studies.

Materials and Methods: The study was performed on 20 male ApoE−/− mice (40 weeks old) divided into groups receiving a standard diet or a Western diet. After anesthesia, PBS perfusion was performed; the heart with the aorta was excised and fixed in 10% formalin; the aorta was cleaned and stained with Oil Red O. The aorta was longitudinally opened, photographed (GelDoc), and quantitatively analyzed using a Python script. Data are presented as mean ± SD; statistical significance was assessed using Student’s t-test at p < 0.05.

Results and Discussion: Thus, the use of the Oil Red O staining method in combination with automated image analysis in Python enables a transition from subjective assessment to precise quantitative measurement of the area of atherosclerotic lesions. This approach increases the reliability of screening therapeutic interventions in experimental models and enhances the statistical power of the study.

Conclusion: In this study, an original approach to morphometric visualization, analysis, and quantitative assessment of atherosclerotic lesions of the vascular wall in an apolipoprotein E–deficient mouse model (ApoE−/−) is proposed and validated for the first time.

Evaluation of pharmacological correction of hypoxic-ischemic encephalopathy sequelae using peptide erythropoietin analogs in a mouse model of mild and moderate hypoxic-ischemic encephalopathy during the late remodeling phase

Vladimir M. Pokrovsky — Thu, 19 Mar 2026 00:00:00 +0300

Introduction: Hypoxic-ischemic encephalopathy (HIE) is an early-life brain injury that remains a leading cause of long-term neurodevelopmental deficits, including cerebral palsy, epilepsy, and cognitive-behavioral impairment. Because persistent neuroinflammation and progressive neurodegeneration contribute to delayed outcomes, targeting EPOR-CD131 with nonerythropoietic erythropoietin derivatives represents a promising strategy to modulate neurovascular unit function and limit secondary injury beyond the acute therapeutic window.

Materials and Methods: CD-1 mice underwent neonatal hypoxia–ischemia at postnatal day 9 (modified Rice–Vannucci) and were stratified 3 h later into mild or moderate injury using laser speckle imaging (RFLSI-ZW), then allocated to treatment groups. Starting on day 7 post-injury, EPO (5000 IU/kg) and Epobis (20 mg/kg) were administered subcutaneously once daily and Ara-290 (30 µg/kg) intraperitoneally twice daily for 7 days, after which exploratory behavior and caffeine-induced neurostimulation responses were evaluated.

Results: In this experimental study, Epobis and Ara-290 normalized mild HIE hyperlocomotion in the open field, whereas EPO mainly improved anxiety-like behavior and only Epobis increased peripheral time toward intact levels. In the caffeine challenge, all treatments reversed paradoxical suppression in mild HIE, and Epobis most effectively reduced caffeine-induced hyperreactivity and restored baseline activity in moderate HIE.

Conclusion: Epobis and Ara-290 normalized mild HIE hyperlocomotion in the open field, whereas Epobis provided the most comprehensive behavioral correction and EPO mainly improved anxiety-related measures. All treatments restored a physiological caffeine response in mild HIE, and Epobis most effectively improved behavior and reduced caffeine-induced hyperreactivity in moderate HIE, supporting further preclinical and subsequent clinical evaluation.

Modeling of cisplatin-induced acute kidney injury and its correction using polydatin

Sun, 31 May 2026 00:00:00 +0300

Introduction: Cisplatin is a key drug used for anticancer therapy. However, its use is often accompanied by the development of acute kidney injury. The aim of this study was to develop an optimal model of cisplatin-induced kidney injury in rats and use it to study the nephroprotective properties of polydatin.

Materials and Methods. The experiment was performed in 100 male Wistar rats. To test the cisplatin-induced acute kidney injury model, seven groups (n=10) were formed. Animals were administered cisplatin intraperitoneally at doses of 1 mg/kg, 5 mg/kg, 10 mg/kg, and 20 mg/kg once or twice on days 1 and 8. Nephrotoxicity was corrected using polydatinat doses of 4 mg/kg and 12 mg/kg. Alpha-tocopherol acetate at a dose of 75 mg/kg was used as a reference drug. The test drug and the reference drug were administered intragastrically daily for 14 days. Nephroprotection was assessed based on the following parameters: creatinine, urea, potassium and sodium ions in the blood serum, glomerular filtration rate, fractional sodium extraction, and renal parenchyma microcirculation.

Results and Discussion. The optimal model of cisplatin-induced acute kidney injury was the one in which cisplatin was administered at a dose of 5 mg/kg on days 1 and 8 of the experiment. This was evidenced by an increase in creatinine level to 124.0 ± 8.6 μmol/L and urea to 20.3 ± 1.2 mmol/L, a decrease in glomerular filtration rate to 0.08 ± 0.01 mL/minand a 2-fold deterioration in microcirculation. Other models were significantly inferior in representativeness. Polydatindemonstrated dose-dependent nephroprotective properties, which was confirmed by improvement in laboratory and instrumental parameters.

Conclusions. The use of cisplatin at a dose of 5 mg/kg intraperitoneally on days 1 and 8 results in optimal modeling of cisplatin-induced acute kidney injury in rats in the experiment. The potential of intragastric use of polydatin for nephroprotection in cisplatin-induced acute kidney injury has been proven in dosages of 4 mg/kg and 12 mg/kg per day for 14 days.

Evaluation of the neuroprotective effects of synthetic erythropoietin derivatives in a mouse model of mild and moderate hypoxic-ischemic encephalopathy

Tue, 17 Mar 2026 00:00:00 +0300

Introduction: Hypoxic-ischemic encephalopathy (HIE) is a leading cause of neonatal brain injury and long-term neurodevelopmental impairment, and current therapies only partially prevent adverse outcomes. Although recombinanterythropoietin is neuroprotective via EPOR–CD131 (βcR), its use is limited by hematopoietic side effects, motivating evaluation of non-hematopoietic peptide analogs such as Ara-290 and Epobis in HIE.

Materials and Methods: Neonatal hypoxia-ischemia was induced in 9-day-old CD-1 mice (n = 204) using a modified Rice–Vannucci model, and animals were stratified into mild and moderate severity groups 3 hours later by laser speckle contrast imaging (RFLSI-ZW) before assignment to treatment. EPO (5000 IU/kg) and Epobis (20 mg/kg) were administered subcutaneously every 24 hours and Ara-290 (30 µg/kg) intraperitoneally every 12 hours for 7 days, after which motor-coordination performance, macroscopic lesion volume, and expression of IL-4, IL-1b, IL-6, and TNF-α were assessed.

Results: In both mild and moderate HIE, Epobis provided the most consistent neuroprotection, improving motor performance and neurological outcomes and being the only treatment to significantly reduce macroscopic lesion volume. EPO produced moderate functional benefits, whereas Ara-290 showed a less stable efficacy profile, with limited or absent effects in several behavioral and morphological endpoints.

Conclusion: In mild HIE, Epobis showed the strongest neuroprotection, improving motor-coordination performance, reducing neurological deficits, and producing the greatest reduction in macroscopic lesion volume, whereas EPO and Ara-290 had only moderate effects. In moderate HIE, Epobis and EPO most consistently improved motor outcomes, but only Epobis significantly reduced lesion volume and neurological symptom severity and produced the most pronounced immunomodulation.