Research Results in Pharmacology

Homozygous mice with mutant protein FUS[1-359] overexpression: Innovative possibilities for ALS treatment

Nikita S. Zhunusov — 2024-12-30

Introduction: This study investigates a mouse model with overexpression of the mutant FUS[1-359] protein, which can be used to evaluate the effectiveness of gene therapy and other pharmacological interventions for amyotrophic lateral sclerosis (ALS). The model enhances the deeper understanding of the mechanisms underlying disease progression and allows for testing new therapeutic strategies.

Materials and Methods: The study utilized tg_hFUS[1-359] animal lines with a transgenic cassette expressing the human mutant FUS[1-359] protein. Animal groups were formed by crossing hemizygous individuals, and analyses were conducted on lifespan, age of disease manifestation, as well as relative copy number and expression levels of the transgenic cassette.

Results and Discussion: The results demonstrated statistically significant differences in the age of onset of initial disease symptoms between homozygous and hemizygous mice. Differences in the copy number of the transgenic insertion were also identified, revealing that homozygous animals exhibited increased expression of the mutant FUS protein in various structures of the central nervous system, consistent with existing literature on ALS pathogenesis.

Conclusion: Mice with hyperexpression of the mutant FUS[1-359] protein represent a promising genetic model for evaluating therapeutic approaches to ALS treatment. This model exhibits clear phenotypic manifestations of the disease and can be utilized for investigating gene therapy methods.

Validation of a mouse model with Apoe gene knockout

Petr R. Lebedev — 2024-12-30

Introduction: The research focuses on the ApoE-/- transgenic mouse model. This model enhances the understanding of the mechanisms underlying disease progression and allows for the testing of new therapeutic strategies.

Materials and Methods: The study employed ApoE (-/-) knockout mice, which were bred to create cohorts of male mice. These males were divided into two equal groups, receiving either a standard diet or a high-fat diet. Analyses included lipid profile assessments, fasting glucose levels, and evaluation of aortic lesion area.

Results and Discussion: The findings revealed statistically significant differences in the ages at which disease symptoms appeared between the groups. The high-fat diet induced hyperlipidemia, leading to accelerated atherosclerosis in the aorta, aligning with existing literature.

Conclusion: ApoE knockout mice represent a promising genetic model for assessing therapeutic approaches to treat cardiovascular diseases. This model exhibits clear phenotypic manifestations of the disease and can be used for pharmacological correction of endothelial dysfunction.

Effectiveness of the tetrapeptide HAEE: an innovative approach to Alzheimer's treatment in experimentation

Evgenii A. Patrakhanov — 2024-12-30

Introduction: Alzheimer’s disease (AD) is the most common neurodegenerative disorder, characterized by a progressive decline in cognitive functions. According to the “Alzheimer’s Disease International”, there were 36 million reported cases of AD worldwide in 2009, with projections suggesting an increase to 66 million by 2030 and 115 million by 2050.

Materials and Methods: The study was conducted on three experimental groups consisting of male APPswe/PS1dE9/Blg mice on a mixed genetic background with C57Bl6/Chg animals. Each group included 10 mice. At the baseline (point 0), peptides and drugs were administered to two groups of animals aged 6 months. The treatments were given circadianly every 48 hours without breaks for one month. Subsequently, at point 1 of the experiment, half of the group (n=5) was selected for further histological analysis of the brain. The remaining half did not receive any treatments for one month before undergoing histological examination. Statistical significance between experimental and control groups was assessed using an unpaired Student’s t-test at p<0.05.

Results and Discussion: Histological analysis indicated the efficacy of the tetrapeptide HAEE at a dosage of 50 mg per kg of mouse weight, showing a significant reduction in amyloid plaques in the cerebral cortex and hippocampus of the mice.

Conclusion: The study supports the proposed hypotheses and suggests further investigation into additional drug groups recommended for Alzheimer’s treatment for comparative studies.

The influence of exogenous recombinant HSP 70 on the alteration of membrane stiffness in hippocampal neurons following the modeling of neonatal hypoxic-ischemic injury in mice

Vladimir M. Pokrovsky — 2024-12-30

Introduction: The use of atomic force microscopy (AFM) to investigate membrane stiffness in neurons provides valuable insights into cellular mechanisms and their alterations in response to various pathophysiological conditions. Heat shock protein HSP 70, a component of the cellular stress response system, plays a role in stabilizing the protein structures of cellular organelles. However, studies examining changes in the stiffness of hippocampal neuronal membranes in its presence, particularly following cerebral circulation disturbances, have not been conducted yet.

Materials and Methods: The study was performed on a mixed culture of hippocampal neurons derived from 9-day-old male CD-1 mice, obtained 24 hours after modeling neonatal hypoxia-ischemia. The following groups were formed: Intact culture; HI culture; HI + rhHSP70 10^{-6 M}; HI + rhHSP70 10^{-8 M}; HI + rhHSP70 10^{-9 M}; HI + rhHSP70 10^{-12 M}, with the substance added in dilutions from an initial dose of 0.1 µg/g. The Young's modulus was measured using force spectroscopy, and maps of local stiffness of various surface areas were generated.

Results and Discussion: The neonatal hypoxia-ischemia model resulted in an 18% increase in the stiffness of the neuronal cell surface compared to the control group (p<0.001). The addition of rhHSP70 at concentrations of 10^{-6 M} and 10^{-8 M} to the HI culture led to an increase in membrane stiffness by 20% (p<0.001) and 3% (p<0.0034), respectively, while dilutions of rhHSP70 at 10^{-9 M} and 10^{-12 M} resulted in a decrease in membrane stiffness by 35% (p<0.001) and 22% (p<0.001) compared to the intact group, respectively. In comparison to such in the neuronal culture group after neonatal hypoxia-ischemia modeling, membrane stiffness with the addition of rhHSP70 at 10^{-8 M}, 10^{-9 M}, and 10^-12 M decreased by 17% (p<0.0004), 65% (p<0.001), and 49% (p<0.001), respectively.

Conclusion: Thus, the addition of rhHSP 70 results in a reduction in membrane stiffness in the mixed culture of hippocampal neurons in mice, compared to the intact culture obtained after neonatal hypoxia-ischemia. The AFM method allows for the assessment of how various molecules, such as heat shock proteins (e.g., rhHSP70), influence the mechanical properties of membranes, which may be critically important for the development of new therapeutic agents.

Pharmacological correction of post-contrast acute kidney injury with a functional product based on fermented grape juice

Olesya V. Shcheblykina — 2024-12-28

Introduction: Post-contrast acute kidney injury (PC-AKI) is a serious complication when takingiodine-containing radiopaque agents. The study is aimed at pharmacological correction of PK-AKIwith a functional product obtained from fermented red grape berry juice with a high content of stilbenoids (trans-resveratrol and its glycoside polydatin).

Materials and Methods: The study was conducted on 36 male CD1 mice, 8-10 weeks old,weighing 25±3 g. The animals were divided into 3 groups (n=12): a group of intact animals; a group with PC-AKI modeling; and a group with PC-AKI modeling and preliminary intragastricadministration of a functional product in a volume of 0.1 mL/10 g per animal. A morphologicalassessment of kidney changes was performed, as well as a blood test for creatinine, urea, andalbumin.

Results: Preliminary administration of the functional product to mice led to a statisticallysignificant decrease in creatinine and urea concentrations to 43.5±1.6 μmol/L and 34.5±1.4μmol/L, respectively; the glomerular filtration rate increased to 66.2±4.1 µL/min and the urea/albumin ratio decreased to 1.9±0.03, with mortality also decreasing to 8.3%.

Conclusion: The preventive use of a functional product based on fermented red berry juice inan amount of 0.1 mL/kg of animal effectively reduces the severity of post-contrast acute kidneyinjury in a mouse model.

Antiparkinsonian activity of selective MAO-B inhibitors of different chemical structures.

Vladimir N. Fedorov — 2024-12-30

Introduction: Parkinson’s disease (PD) is a chronic neurodegenerative disease of the central nervous system, the pathogenesis of which is associated with the death of dopaminergic neurons of the midbrain substantia nigra. The mainstay of therapy for PD is levodopa. However, in the initial stages of PD or in case of levodopa intolerance, MAO B inhibitors are used. Purpose of the study was to determine antiparkinsonian activity of newly synthesized selective MAO-B inhibitors in vivo on the model of experimental parkinsonism in white mice.

Materials and Methods: Parkinsonian syndrome in mice was modeled by systemic administration of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). As the criteria for its evaluation, we used the determination of muscle rigidity severity by step length of mice and evaluation of oligokinesia severity and emotional and mental disturbances in the open field test. A total of 9 candidate compounds were studied, assigned with laboratory codes S1-S5, S9, S10, S14, and S15. Rasagiline was used as a comparison drug.

Results and Discussion: Of the 9 compounds used at a dosage of 2mg/kg, the degree of rigidity was significantly reduced by compounds S1, S9, and S15; locomotor activity was restored to the level reached through rasagiline only by compound S9; the decline in exploratory activity was prevented only by S9 and to some extent by S5.

Conclusion: Only compound S9 having benzenesulfonamide chemotype showed significant therapeutic potential in a model of experimental parkinsonism and only in relation to it additional studies can be planned.

The mitigating effect of a topical preparation of amlodipine on imiquimod-induced psoriasis-like lesion in mice

Zainab A. Mahdi — 2024-12-30

Introduction: Psoriasis is a chronic inflammatory skin disease depicted by deforming, inconsistent penetrations, and repeated proliferative and inflammatory skin ailment. Local treatment with topical agents is one of the substantial modalities in the remediation of this ailment. Efforts are directed toward the development of novel efficacious topical medicaments. The study examined the potential anti-psoriatic effect of topical amlodipine as a 5% ointment in mice based on observational, histopathological, and biomarker outcomes.

Materials and Methods: Forty-five male Swiss Albino mice were randomly allocated into five distinct groups (I-V), with 9 mice in each group (n=9) that underwent shaving of the dorsal hair. In Group I; animals served as control, while animals of other groups received imiquimod (IMQ) on their shaved backs for six consecutive days to induce psoriasis. Groups from III to V received continuous application of IMQ following day 6 along with a vehicle, clobetasol, andamlodipine 5% ointment for 8 consecutive days. During the timeline of the experiment, observation changes were assessed daily, followed by animal euthanasia and sample collection for biochemical and histopathological evaluation.

Results: Topical amlodipine ointment markedly reduced the inflammatory signs of psoriatic lesions, and histopathological inspection confirmed these findings. The levels of interleukin-17 (IL-17), tumor necrosis factor-alpha (TNF-a), and vascular endothelial growth factor (VEGF) were significantly ameliorated by topical amlodipine Ointment in comparison with the untreated imiquimod-induced psoriatic mice group.

Conclusion: The study concludes that topical amlodipine showed an anti-psoriatic effect comparable to that of topical clobetasol owing to antioxidant and anti-inflammatory effects making amlodipine a promising agent in the future of psoriasis management as an adjuvant treatment to the standard.

The evaluation of pharmacokinetic parameters of 4-(5-methyl-1,3,4-oxadiazole-2-yl)-benzenesulfonamide and its metabolites in rat plasma

Alexander L. Khokhlov — 2024-12-28

Introduction: 4-(5-methyl-1,3,4-oxadiazole-2-yl)-benzenesulfonamide (ODASA) is a new pharmacologically activecompound, which is capable of reducing intraocular pressure by inhibiting carbonic anhydrase II. It is necessary to calculate the pharmacokinetic parameters of this compound and its metabolites in blood plasma of laboratory animals during a preclinical study. Aim: Evaluation of pharmacokinetic parameters of ODASA and its metabolites in rat bloodplasma after instillation of ocular suspension and its intraperitoneal administration.

Materials and Methods: The study was conducted on 2 groups of Wistar rats, each including 6 individuals. The firstgroup underwent instillation of 1% ocular suspension of ODASA of 20 µL into each eye (1.6 mg/kg). The drug was injected intraperitoneally at the same dose to the second group. Blood sampling was performed before the administration,0.5h, 1h, 2h, 4h, 6h, 8h, 12h, 24h, 48h, 72 h, 96h, 120h, 144h, 192h, 240h, 288h after the administration. Plasma was immediately stabilized by 10% solution of ascorbic acid and frozen to a temperature no higher than -70°C. The sampleswere analyzed using the HPLC-MS/MS method. Chromatographic separation was performed on a Kinetex Phenyl Hexylcolumn (50*4.6 mm, 2.6 microns) in a gradient mode. Detection was carried out in the MRM mode using electrosprayionization.

Results: The developed method was validated in the range of 2-2000 ng/mL for ODASA and 4-[5-(hydroxymethyl)-1,3,4-oxadiazole-2-yl]-benzenesulfonamide (M1) and 0.5 -500.0 ng/mL for N-hydroxy-4-(5-methyl-1,3,4-oxadiazole-2-yl)-benzenesulfonamide (M2). The size of the maximum plasma concentration after instillation of ODASA into eyes was 349.85±62.50 ng/mL, in M1 – 30.91±6.00 ng/mL and in M2 – 2.70±0.62 ng/mL (M±SEM). The half-life time of ODASA after ocular administration was 46.4±3.8 h, M1 – 70.0±14.3 h, and M2 - 36.5±15.2 h (M±SEM). The relative bioavailability of ODASA compared with injection was 81.03%.

Conclusion: The performed validation of the method guaranteed the accuracy of the obtained results. The activesubstance has a high relative bioavailability after instillation into eyes. M1 is a major metabolite, and M2 is a minormetabolite. The studied compounds had a long half-life.

CAR-T cells: prospective genetic engineering approach to orchestrate solid tumor in lung cancer

Angeline Felisca Tanujaya — 2024-12-30

Introduction: Various strategies, starting with surgery, systemic radiation therapy with cytotoxic chemotherapy, and immunotherapy, have been carried out to address mortality from lung cancer. None of these therapies has showed remarkably successful treatments for lung cancer. Developing technology leads to innovation of cancer-targeted therapy as an ideal strategy. Chimeric antigen receptor T cells (CAR-T cells) are one of the novel immunotherapy approaches in genetic engineering. CAR-T therapy has showed promising results as the cancer-targeted therapy.

Methods: This review identifies relevant research by the keywords “Lung Cancer”, “CAR-T for Lung Cancer”, “CAR-T for solid tumor”, “CAR-T for SCLC” or “CAR-T for NSCLC”. The articles are extracted from Pubmed, Web of Science, Scopus, Science Direct, and Google Scholar. Data are shown based on discussions of the modification receptor, co-stimulator, and CAR-T cells.

Results: CAR-T acts to overcome lung cancer by targeting several receptors, such as mesothelin and c-Met. Several generations have also evolved to more specific and effective therapy. CAR-T modifications like BBIR, SUPRA CAR, and UIR have also been developed to minimize side effects.

Conclusion: CAR-T developments are not limited to CAR modification, but include its components and structure.

Diagnosis and treatment of arterial stiffness, pulmonary hypertension, diastolic cardiac dysfunction against the background of ischaemic heart disease in comorbid patients

Nadezhda N. Pribylova — 2024-12-30

Introduction: In the context of cardiology and neurology, special attention is paid to the problems of cardiovascular and cerebrovascular pathologies, often caused by vascular dysfunction. Haemodynamic parameters, in particular arterial stiffness (AS), and epicardial fat thickness (EFT), play a significant role in the assessment of cardiovascular health. The aim of the study was to evaluate the correlation between arterial stiffness, pulmonary hypertension (PH), epicardial fat thickness and diastolic myocardial dysfunction in comorbid patients.

Materials and Methods: The comparative study was conducted in three groups of patients with the most frequent comorbid pathology. The first group (n=75) included patients with ischaemic heart disease (IHD), arterial hypertension stage II-III and chronic obstructive pulmonary disease (COPD) stage II-III. The second group (n=50) consisted of patients with IHD and arterial hypertension without COPD. The third group (n=33) included patients with IHD without comorbidities.

Results: The study revealed a significant correlation between indices of AS, blood pressure (BP), EFT and PH in patients with comorbid conditions. The study found that the addition of the combined antihypertensive drug amlodipine-perindopril to standard therapy contributed to normalization of AS, PH and BP during three months of treatment. There was also a significant improvement in the patients’ quality of life, reduction of dyspnoea and heart failure symptoms.

Conclusion: The obtained data confirm the presence of correlation between AS, BP, PH and EFT in patients with IHD, AH and COPD, which may serve as indicators of comorbid diseases. The use of combined antihypertensive drug amlodipine-perindopril seems reasonable and scientifically justified, especially after coronary stenting.