Background. Osteoarthritis (OA) is characterized by heterogeneity of clinical manifestations and, in some cases, a severe progressive course. In this regard, it is important to identify new molecular targets for the treatment of the disease.

Aim. To determine the role of pathological immune processes, specific genetic and epigenetic changes in OA, identification of OA-specific microRNAs and potential targets for targeted therapy.

Materials and methods. To prepare the review, scientific platforms PubMed, Scopus, ResearchGate, RSCI were used to search for information. The search words and phrases were: “osteoarthritis genes meta-analysis”, “osteoarthritis genes”, “miRNAs osteoarthritis”.

Results. Data were obtained on the involvement of pathological immune reactions in the mechanism of OA with changes in the expression of 34 specific genes involved in the functioning of the immune system by immune cells infiltrating joints. Clinical studies have determined the association of allelic variants of C5AR1, FCGR2B, HLA-DR2, HLA-DR5, IL1B, IL1RN, IL4R, IL6, IL10, IL17, TYROBP, TLR3, TLR4, TLR7, TLR9, TLR10 genes, involved in the regulation of immune system functioning. Changes in the expression of 11 specific microRNAs involved in inflammatory and degenerative processes in OA were identified.

Conclusion. Molecular genetic studies make it possible to find new markers of pathological immune reactions in OA, the presence of which in patients can be used to determine methods of treating the disease to prevent rapid progression of the disease, as well as to design targeted therapy. An important role of disturbances in the expression of genes involved in the functioning of the immune system in the pathogenesis of the disease was identified. MicroRNAs associated with OA involved in the pathogenesis of immune changes may become promising tools for targeted therapy of OA. Analysis of the reviewed materials indicates that the use of microRNAs that affect retroelements involved in the pathogenesis of OA can become the basis not only for suppressing the progression of the pathology, but also for slowing down the aging process.

Background. One of the approaches to expand natural killer (NK) and NKT cells for subsequent CAR (chimeric antigen receptor) therapy is cultivation in the presence of certain cytokines and/or feeder cells.

Aim. To evaluate the effect of combinations of interleukin (IL): IL-2 + IL-15, IL-2 + IL-15 + IL-21 or IL-2 alone on NK and NKT cells during long-term cultivation in the presence of autologous feeder cells, which were non-irradiated T lymphocytes activated by monoclonal antibodies (mAbs).

Materials and methods. Peripheral blood and leukocyte-platelet concentrate were used as a source of peripheral blood mononuclear cell. After peripheral blood mononuclear cell isolation, the lymphocyte culture was activated using adsorbed mAbs to CD3 and CD28 receptors. Then, activated lymphocytes were cultured in the presence of IL-2 + IL-15, IL-2 + IL-15 + IL-21, or IL-2 alone, periodically assessing the increase in the total number of cells, expression of surface markers, and cytotoxic activity.

Results. Using the combination of IL-2 + IL-15 + IL-21, the greatest increase in cells and maximum cytotoxicity were observed; the proportion of NKT cells (CD3⁺CD56⁺) by the end of the cultivation was also the highest compared to other cultivation modes and amounted to 59 %. In the presence of a combination of IL-2 and IL-15, lower cytotoxicity of the cultured lymphocytes was determined compared to other experimental options, which may be due to exhaustion of the cell population. Based on the literature data, the addition of IL-21 to the culture medium probably neutralizes this negative effect of IL-15.

Conclusion. Based on the dynamics of proliferation, receptor expression and cytotoxicity of cultured lymphocytes, it can be concluded that the cytokine combination consisting of IL-2, IL-15 and IL-21 is most effective for enriching the NKT cell population upon stimulation with mAb and in the presence of autologous activated T lymphocytes. For the most effective generation of the NK cell population, it is necessary to use other feeder cells, probably of tumor origin, or other expansion protocols.

Background. Medicinal plants have a high chemopreventive potential in oncology. The structural diversity of plant biologically active substances: phenolic compounds (simple phenols, flavonoids, phenolic acids, tannins, etc.), terpenoids (mono-, diterpenoids, triterpene saponins, etc.), essential oils, for example, results in a wide range of antitumor activity. Complex herbal preparations are more effective and safe due to their multi-target effects on various regulatory mechanisms in combination with relatively low toxicity. The development of effective natural pharmaceutical compositions based on the structural diversity of biologically active substances seems relevant.

Aim. Number plant extracts antiproliferative effect against CaOv human ovarian adenocarcinoma cells investigation and potential phytocomponents selection for the novel antitumor pharmaceutical composition development.

Materials and methods. CaOv human ovarian adenocarcinoma cell line was used as a test model. The antiproliferative effect of plant extracts was measured by the radiometric method based on the ³H-thymidine incorporation into CaOv cells DNA. Half maximal inhibitory concentration (IC₅₀) was calculated using probit analysis. The selection of vegetable oil as an optimal extraction agent was carried out on C57BL/6 male mice based on the wound healing effect.

Results. The antiproliferative activity of 27 medical plant extracts was studied. The 14 most effective ones in these conditions were selected based on the IC₅₀. Linseed oil was chosen as an optimal extraction agent considered the wound-healing effect of individual vegetable oils (linseed, sunflower, corn), as well as the corresponding oil extracts of the phytocomposition.

Conclusion. The selected phytocomponents can be used as a foundation for an optimal compound development of a new pharmaceutical composition aimed at chemoprevention in cancer care. The choice of linseed vegetable oil as an experimental phytocomposition extractant is justified.

Background. Small or short double-stranded interfering RNAs (siRNAs, small interfering RNAs) 20–25 nucleotides long are known to be able to target block uncontrolled malignant proliferation. As target genes, apoptosis inhibitors are considered, including the cellular glycoprotein CD47 (cluster of differentiation 47), and genes of the replicative complex that regulate the cell cycle in the S phase. This determines the relevance of the study in tumor models of siRNAs aimed at these targets as adjuvants.

Aim. To evaluate the antiproliferative effects of novel siRNAs as adjuvants for immune-/chemotherapy in human colorectal and renal cancer models.

Materials and methods. SiRNA/antiCD47 and two-component siRNA antiMSM4/antiLIVIN were developed at the Research Centre for Medical Genetics and studied in lipid dispersion for intravenous (IV) administration. Preclinical models – subcutaneous xenographs of RTK-8 colon cancer and human kidney cancer Rpoch-1/CD47, Balb/c nude mice were obtained from the N.N. Blokhin National Medical Research Center of Oncology. In the adjuvant mode, siRNA/antiCD47 was studied in combination with activated human macrophages (AM), siRNA antiMSM4/antiLIVIN (1:1) – with cyclic-dependent cytostatic oxaliplatin (OXP). Administration regimens are justified earlier. Efficacy parameters and criteria (treatment/control (T/C) ≤42 %), tolerability of effects and statistical analysis at p <0.05 are standard for experimental cancer therapy. Laboratory manipulations are regulated by the current recommendations of the Ministry of Health of the Russia.

Results. The siRNA/anti-CD47 + AM regimen was practically ineffective at Rpoch-1/CD47, T/C = 45 % (p >0.05). The antiMCM4/antiLIVIN + 24 h siRNA regimen on an OXP-insensitive RTK-8 showed a significant adjuvant effect against cytostatic, T/C = 33–21 % versus T/C_min = 49 % (p ≤0.05). Both combinations were tolerable.

Conclusion. Preclinical study showed the controversy of the assumption about the possibility of adjuvant use of siRNA/antiCD47 with AM and the promise of antiMSM4/antiLIVIN siRNA on low-sensitivity to cycle-dependent OXR in human colon cancer with the possibility of cell cycle synchronization.

Background. Neurotoxicity is a side effect of anthracycline antibiotics that has been identified during clinical use. while this type of toxicity may not be limiting, it can significantly affect the quality of life for patients. In the Gause Institute of New Antibiotics an antitumor compound called anthrafuran has developed, that is similar in structure to anthracyclines. This compound has shown high activity in experiments using mouse models of transplanted tumors when administered orally. Anthrafuran has the ability to penetrate the blood-brain barrier, so a study of its neurotoxicity was previously conducted at the maximum tolerated dose.

Aim. To experimentally evaluate the neurotoxicity of anthrafuran when it is administered orally at both a therapeutic dose and three times the therapeutic dose.

Materials and methods. Female Albino rats were used in the experiment. The animals were kept under conditions accordance to GOST 33044–2014 “Principles of good laboratory practice”. Anthrafuran substance was administered orally as a 1,2 % solution in 5 % glucose for injection at doses of 20 and 60 mg/kg once. Motor and research activity of the animals was evaluated in an Open Field test setting 4 hours, one day, and one month after administration. To detect cognitive dysfunction, rats were trained in a T-maze with food reward 3–5 days after drug administration.

Results. Administration of the drug at a therapeutic dose of 20 mg/kg did not cause any abnormal behavior in animals in the Open Field or affect the ability to learn in the T-maze. However, at a dose three times higher than the therapeutic dose (60 mg/kg), anthrafuran decreased the research activity of rats in the Open Field 4 and 24 hours after administration and inhibited the ability to acquire learning in T-maze.

Conclusion. The use of anthrafuran in a therapeutic dose did not cause pronounced neurotoxic reactions. In order to further promote the drug, it is necessary to conduct an in-depth study on the effect of the substance and dosage forms on the behavioral responses and cognitive abilities of rats.

Background. Currently, various approaches can be used to increase the solubility and dissolution rate of poorly water-soluble pharmaceutical substances, such as salt formation, solubilization with co-solvents, particle size reduction, or preparation of solid dispersions. A promising and relevant area in pharmaceutical science is the production of solid dispersions. Polyvinylpyrrolidone and polyethyleneglycols of various molecular weights are most often used as carrier polymers in the production of solid dispersions.

Aim. Analysis of desloratadine solid dispersions by physicochemical and thermal methods in order to substantiate the most optimal composition and technology for obtaining solid dispersions.

Materials and methods. Solid dispersions of desloratadine with polyethyleneglycol-1500, polyethyleneglycol-4000, polyethyleneglycol-6000, polyvinylpyrrolidone-10000 as carriers in the ratios of 1:1, 1:2, 1:5 were used as objects of study. To determine the morphological features of the obtained samples, scanning electron microscopy was used on a JSM-6380LV device (JEOL, Japan). IR spectroscopy was performed on a Vertex-70 device (Bruker Optik GmbH, Germany), in the mid-IR region of 4000–400 cm^–1 using the total internal reflection method. In order to study the crystal structure of solid dispersions with polymer carriers, X-ray phase analysis was performed using the powder X-ray diffractometry method on a DRON device. Studies by the differential scanning calorimetry (DSC) method were carried out on a synchronous thermal analysis device model STA 449 F3 (Netzsch, Germany).

Results. IR spectra of desloratadine solid dispersions demonstrated fluctuations in the areas corresponding to the functional groups of the pharmaceutical substance and polymers. The X-ray diffraction pattern of samples of desloratadine solid dispersions with polymers shows a loss of the crystalline structure of the pharmaceutical substance. when conducting differential scanning calorimetry, the lowest value of specific heat of complexation was found for solid dispersions of desloratadine with polyethyleneglycol –1500 and polyethyleneglycol –6000.

Conclusion. The conducted studies showed that the optimal polymer for obtaining solid dispersions is polyethyleneglycol-1500.

Background. Currently, the issues of skin and mucous membrane wound therapy are becoming increasingly clear. One of the most modern and popular dosage forms of wound healing agents is a gel, which has undeniable pharmacological and technological advantages. L-arginine, which is a substrate for nitric oxide (II) (NO (II) synthases), which are involved in the formation of NO (II), was used as the main active ingredient in the developed dosage form.

Aim. Selection of the optimal gelling agent and development of the composition of a soft dosage form used for healing wounds of the skin and mucous membranes.

Materials and methods. Three model samples of gels made using potentially promising gelling agents were selected as objects of study: carbopol 940, sodium carboxymethyl cellulose, methyl cellulose. Rheological characteristics of the samples were studied by rotational viscometry on a Fungilab Premium viscometer (Spain) using a “cylinder in cylinder” measuring system. To study the biodegradation of the model sample, a FADT-1202A dissolution tester (FOCS Co., Ltd, China) equipped with a paddle stirrer was used. The degree of arginine release from the samples was estimated by the Kruvchinsky equilibrium dialysis method.

Results. As a result of studying the structural and mechanical properties of the model samples, it was revealed that methylcellulose in combination with L-arginine allows creating a gel composition with the greatest stability. when studying the biodegradation of the methylcellulose-based composition, it was found that the gel is capable of being on the oral mucosa for no more than 60 minutes. Then, using the Kruvchinsky equilibrium dialysis method during this period of time, the nature of the L-arginine release from the developed composition was established.

Conclusion. The conducted studies substantiated the choice of the wound-healing gel base. Based on the obtained results, methylcellulose was chosen as the optimal gelling agent for the production of a soft medicinal form with a wound-healing effect.

Background. The glycosylated derivative of indolo[2,3-a]carbazole LCS-1269, an DNA intercalator and a chromatin remodulator, synthesized in N.N. Blokhin National Medical Research Center of Oncology (N.N. Blokhin NMRCO), is of great interest for the treatment of malignant tumors. To carry out pharmacokinetic studies of LCS-1269 dosage forms in vivo, it is necessary to develop a method for LCS-1269 assay in biological samples.

Aim. To develop and validate an analytical technique using high-performance liquid chromatography with ultraviolet detection (HPLC-UV) for LCS-1269 pharmacokinetic studies.

Materials and methods. The study used the LCS-1269 substance containing 99.4 % of the active component, blood plasma and liver of healthy cyclic immunocompetent laboratory mice hybrid (C₅₇Bl/6J×DBA2) F₁ of N.N. Blokhin NMRCO breeding. Acetonitrile and sodium chloride were used to prepare biological samples. Reversed-phase HPLC-UV method in the gradient regimen was used for LCS-1269 assay. Statistical processing of the obtained results was carried out in Microsoft Office Excel 2010 software.

Results. The technique based on cooled acetonitrile deproteinization was developed for blood plasma sample preparation; liver samples were prepared using combined cooled acetonitrile and sodium chloride deproteinization. The developed technique validation confirmed its specificity, linearity (r >0.99), correctness (response >80 % at low concentrations) and precision (relative standard deviation <3.5 %) in the entire analytical range; LCS-1269 transfer effect wasn’t observed. It was found that the analyzed samples are stable for 24 hours at room temperature, and the model mixtures are stable during a month when stored at temperatures less than –18 °C.

Conclusion. A bioanalytical technique for LCS-1269 assay in the blood plasma and liver of animals using HPLC-UV has been developed. Has been shown that the technique meets the requirements of the Eurasian Economic Union for the validation of biological samples analysis.

Background. Gemcitabine is used in oncology for the treatment of various solid tumors, including lung cancer. Gemcitabine exhibits high therapeutic efficacy by inhibiting DNA synthesis and inducing apoptosis in tumor cells. However, the drug has drawbacks, such as a short half-life and rapid metabolic degradation, necessitating frequent administration of high doses, which increases the risk of side effects. To reduce toxicity and enhance therapeutic efficacy, we developed a liposomal formulation of gemcitabine.

Aim. To compare the cytotoxic activity of liposomal gemcitabine and gemcitabine solution on the A549 lung cancer cell line.

Materials and methods. The cytotoxicity of liposomal gemcitabine and gemcitabine solution was evaluated on the human lung adenocarcinoma cell line A549. Cells were cultured in the presence of liposomal gemcitabine and gemcitabine solution in vitro for 72 hours, and the cytotoxicity of the drugs was assessed using the MTT assay.

Results. Liposomal gemcitabine demonstrated higher cytotoxicity compared to free gemcitabine. Half maximal inhibitory concentration value for liposomal gemcitabine was 0.47 μM, while that for free gemcitabine was 9.6 μM.

Conclusion. The liposomal form of gemcitabine exhibits significantly higher cytotoxic activity compared to the free gemcitabine solution against the A549 lung cancer cell line. Incorporation of gemcitabine into liposomes improved drug penetration into tumor cells and protected it from premature degradation.