Vol 22, No 6 (2019)

The pot marigold flowers (Calendula officinalis L.) are widely used in worldwide medical practice. The extensive spectrum of pharmacological activity of Calendula flowers (anti-inflammatory, regenerating, antimicrobial, cholagogue, expectorant properties) is based on the presence of various classes of biologically active substances, namely carotenoids, flavonoids (glycosides of kaempferol, quercetin and isorhamnetin), saponins. This factor makes Calendula a highly promising resource of new herbal medicines. Nevertheless, in the literature there are conflicting data on the chemical composition of medicinal marigolds, which require an additional study of the chemical composition of pot marigold flowers. The aim of this study is thephytochemical study of compounds isolated from flowers of pot marigold, cultivated in the Samara region. Materials and methods. The object of the study was the flowers of marigolds, cultivated on the pharmacopoeial site of the Botanical Garden of Samara University. The chemical structure of the substances was established using the data of 1H-NMR, 13C-NMR-spectroscopy, UV spectroscopy and mass spectrometry. Results.Calenduloside K, which is a new natural compound and having the structure: 3-O - [(1 → 4) -ß-D-glucopyranosyl- (1→ 6)-ß-D-glucuronopyranosyl] -ß-D-glucopyranoside of oleanolic acid, was isolated using column chromatography from the Сalendula flowers sort «Kalta» cultivated in the Samara region), as well as 3-O-ß-D-glucopyranoside of isorhamnetin, 3-O-a-L-rhamnopyranoside of isorhaminein and 3-O-p-coumaroylquinic acid. In addition, a diagnostically significant and dominant flavonoid narcissin was isolated from the Сalendula flowers. As a result of the carried out researches, saponin (calenduloside K), flavonoids (narcissin, 3-O-ß-D-glucopyranoside of isorhaminein, 3-O-a-L-rhamnopyranoside of isorhamnetin) and phenylpropanoids (3-O-p-coumaroylquinic acid). It was determined that flavonoid narcissin (3-O-rutinoside of isorhamnetin) is the dominant and diagnostically significant flavonoid of Calendula flowers. In our opinion, it is advisable to determine the authenticity of flowers of Calendula officinalis by detection by TLC of narcissin, rather than rutin, as provided by the current regulatory documentation. Conclusion. As a result of the carried out researches, saponin (calenduloside K), flavonoids (narcissin, 3-O-ß-D-glucopyranoside of isorhaminein, 3-O-a-L-rhamnopyranoside of isorhamnetin) and phenylpropanoids (3-O-p-coumaroylquinic acid). It was determined that the analysis of the raw material «Calendula officinalis flowers» is expedient to be carried out by TLC method by detection of narcissin, taking into account its specificity, and also the fact that this flavonoid is the dominant one in this plant.

Aim purpose. The search for optimal conditions for isolating felodipine, its purification and the development of methods for determining felodipine in biological material. Methods. The object of study is felodipine (3-ethyl-5-methyl-4- (2,3-dichlorophenyl) -2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylate). Conducted research on the comparative isolation of felodipine from the biomaterial with fluids of organic nature, water and aqueous solutions of various reactions. To perform the experiments, model mixtures of felodipine and crushed liver tissue (particle size 0.2-0.5 cm) with a content of 0.05% of the substance were prepared. Acetone has been proposed as an insulating agent for the recovery of felodipine from biological fluids. For identification and quantification of felodipine in extracts from biological material have been proposed methods of thin layer chromatography (TLC), spectrophotometry, and chromatography - mass spectrometry (GC - MS) Results. It has been established that optimal conditions for the extraction of felodipine from biological material are achieved already with double infusion of a biological object with an insulating agent, if the mass ratio of the insulating liquid and the biomaterial at each stage of infusion is at least 2 : 1, and the duration of infusion is at least 30 minutes. Optimal purification conditions for felodipine were achieved in the macrocolumn of the Silaborus S-18 sorbent 30 μm in height 150 mm and 10 mm in diameter with elution of the substance with acetonitrile-water polar eluant (7 : 3). When the content of felodipine in the amount of 25 mg in 25 g of biological material, the developed method allows to determine 83.91-85.55% in the liver, 83.82-85.63% in the putrefactive liver of this substance. The openable minimum by the method (100 g of a biological object) is: 0.150 mg of the substance in the liver, 0.220 mg in the rotten liver. Conclusion. Thus, the technique can be applied in the practice of chemical-toxicological analysis to study the tissues of fresh and putrefactive corpse organs for the presence of felodipine in them.

Relevance. The results of molecular biological studies served as the basis for conducting a clinical study on the possibility of using antiangiogenic drugs in the treatment of bone sarcomas. Their mechanism of action is to inhibit tyrosine kinases involved in tumor growth, including inhibiting the binding of a key activator of neoangiogenesis of vascular endothelial growth factor (VEGF) with its type 1 and 2 receptors (VEGFR1, VEGFR2), which leads to a decrease in vascularization and inhibition tumor growth. In addition to VEGF, other biologically active substances, including the interleukin-16 cytokine (IL-16), are also involved in tumor neoangiogenesis activators. Goal. Comparative study of the initial levels of VEGFR1 and VEGFR2 receptors in the serum of patients with malignant, borderline and benign bone neoplasms with significant levels of IL-16 and their role in the bone sarcomas prognosis. Material and methods. We examined 138 patients with bone tumors (malignant - 106; benign - 10; borderline (giant cell tumor) - 22). The markers were studied by an enzyme immunoassay method in the blood serum of patients before the specific treatment with Biosource (USA) reagents - IL-16 and "R & D" (USA) - VEGFR1 and VEGFR2. Statistical processing of the results was performed using the program "Statistica 7.0". Results. Significant levels of IL-16 were detected in serum samples of 93% of patients with bone neoplasms. Differences in the content of IL-16 in the serum with regard to the morphological structure of the tumor was not detected. In typical osteosarcoma and Ewing's sarcoma, serum levels of VEGFR1 and VEGFR2 are significantly higher than in chondrosarcoma (p = 0.0003 and p = 0.00009; and p = 0.032 and p = 0.005, respectively). The serum levels of VEGFR1 and VEGFR2 in patients with bone tumors were significantly higher at baseline levels of IL-16 <33.0 pg / ml than in patients with high levels of IL-16 > 33.0 pg / ml (p = 0.027 and p = 0.026 respectively). A significant direct relationship was found between serum levels of VEGFR1 and VEGFR2 (r = 0.34, p = 0.002). The overall 5-year survival in the entire group was 56%. The overall 5-year survival in the entire group was 56%. With a combination of IL-16 <33 pg / ml and VEGFR2 <11.3 ng / ml, the overall 5-year survival was 85%, whereas at high IL-16 values >33 pg / ml and low VEGFR2 < 11.3 ng / ml - it was equal to 25%. The overall 5-year survival rate in the presence of all high values of IL-16, VEGFR1, VEGFR2 in the blood was 67%, for all low values of marker - 78%. It should be noted that with a combination of levels of IL-16> 33.0 pg / ml, VEGFR1 <108.5 pg / ml, VEGFR2 >11.3 ng / ml and IL-16 <33.0 pg / ml, VEGFR1 >108, 5 pg / ml, VEGFR2 <11.3 ng / ml -the indicators of total 5-year survival were maximum - 100%. The worst outcomes of overall 5-year survival (0%) were observed with a combination of IL-16 >33.0 pg / ml, VEGFR1 >108.5 pg / ml, VEGFR2 < 11.3 ng / ml and IL-16 <33, 0 pg / ml, VEGFR1 <108.5 pg / ml, VEGFR2 >11.3 ng / ml. Conclusion. The results indicate the promise of conducting studies on the content of IL-16, VEGFR1 and VEGFR2 in the peripheral blood of patients with bone neoplasms for studying the possibility of their use as prognostic markers.

Purpose of the study. A comparative study of the anticonvulsant activity of microencapsulated forms of midazolame on the model of pentilenetetrazol seizures. Materials and methods. Anticonvulsant activity of midazolam and microencapsulated samples of midazolam based on alginate and polylactide-co-glicolide was evaluated on white outbred male mice on a model of convulsive syndrome caused by the intramuscular administration of pentylenetetrazole at a dose of 1 LD99. Results. It has been established that the inclusion of midazolam in polymer particles based on sodium alginate with a ratio of midazolam and polymer 1:9 contributes to an increase in the duration of the anticonvulsant effect and the protective action of prophylactic administration 5-30 minutes before poisoning. Findings. The advantage of sodium alginate-based polymer particles over polylactide-co-glicolide-based particles with respect to the anticonvulsant and protective efficacy of midazolam in experiments on mice is shown.