Chondroitin sulfate is used for osteoarthritis combination therapy. It should be taken into account that the structure and properties of polysaccharides included in chondroitin sulfate, as well as the raw materials used for its production, have a significant effect on its absorption, bioavailability, and, as a consequence, on the safety and efficacy of orally administered products.The aim of the study was to assess toxic properties, local irritant effect, immunotoxicity, basic pharmacokinetic parameters, and therapeutic efficacy of the new Chondroitin sulfate product (produced by Federal State Unitary Enterprise “Moscow Endocrine Plant”, Russia) as compared to Structum (produced by “Pierre Fabre Medicament Production”, France).Materials and methods: White Giant rabbits were used in the experiments. Toxicity, immunotoxicity and local irritation effects of the products were assessed following daily oral administration at the dose of 168 mg/kg (about 6 Maximum Recommended Therapeutic Doses) to male and female rabbits for 28 days. The follow-up period was 14 days. The pharmacokinetic study included blood sampling on days 1‒2 of the experiment, complete blood count and blood chemistry tests were performed on days 28 and 43. After killing the animals, pathomorphological and histological examinations were performed on their organs and tissues. Therapeutic efficacy was studied in an osteoarthritis model made by cruciate ligament transaction in rabbits. The animals received therapy at doses of 16.8 mg/kg, 33.6 mg/kg, and 67.2 mg/kg for 56 days starting from day 8 after the pathology induction.Results: the medicines had no toxic, local irritant, or immunotoxic effect. The NOAEL was established at 168 mg/kg. The study demonstrated the comparability of the pharmacokinetic profiles of the studied products following single oral administration. The maximum concentration of the active ingredient (Cmax = 79 ± 6 μg/mL—Chondroitin sulfate; Cmax = 71 ± 4 μg/mL— Structum) in blood plasma was observed within 3–4 hours after administration. A decrease in the severity of cartilage structural damage was observed for the doses of 33.6 mg/kg and 67.2 mg/kg. The results of quantitative determination of sulfated glycosaminoglicans in the proteoglycans of the cartilage articular surface in the animals with osteoarthritis demonstrated an increase in the level of sulfated glycosaminoglicans in the groups treated with the maximum doses of the studied products, as compared to the other groups.Conclusions: the obtained data confirm that the test product has a favourable safety profile, and therapeutic (chondroprotective) effect. All the tested properties of Chondroitin sulfate were comparable to those of Structum.

Introduction. The pharmacokinetic profiling of active compounds is necessary for drug development and application. Comlex extract of biologically active compounds was isolated from gonads of green sea urchin Strongylocentrotus droebachiensis from Barents Sea. It contains fatty acids, carotenoids and tocopherols and have inhibition activity to ensyme dypeptydylpeptidase IV (DPP-4). Traditional approaches to pharmacokinetic study based on chromatographic methods are not effective for such complex mixtures because they had not enough selectivity and sensitivity. Methods of immunoassay for special enzyme, bioassay, etc. may be used as an alternative way.Aim. The aim: to find approach to pharmacokinetic study of sea urchin gonads extract in rabbits after a three doses oral administration.Materials and methods. Various spectroscopic and chromatographic (HPLC and TLC) methods were used for chemical characteristic of sea urchin gonads extract ant different target analytes quantification in biosamples. The pharmacokinetic of sea urchin gonads extract was studied after single dose oral administration to male rabbits. The correlation between concentration of sea urchin gonads extract in the blood plasma and its biological activity marked by DPP-4 activity was established. The activity of DPP-4 was determined by the chromogenic optical method.Results and discussion. Main groups of sea urchin gonads extract chemical compounds were characterized. Total peptides content was 15-22 %; a-tocopherol content - 0.05-0.15 %, total tocopherols content - 0.23-0.38 %; total carotenoid content - 0.005-0.07 %; total fatty acids content calculated to linolenic acid - 11,03 to 12,74 %. After sea urchin gonads extract chemical composition results it was established that concentrations of identified individual biologically active compound are low, there is no dominant compound or group of compounds responsible for pharmacological activity of sea urchin gonads extract which may be unambiguously chosen as target analyte for its pharmacokinetic study. Approach based on correlation of special marker activity (DPP-4) and extract concentration appears to be the best for pharmacokinetic study of complex extract from green sea urchin gonads. The method for the quantitative determination of sea urchin gonads extract in the blood plasma of rabbits by its effect on DPP-4 activity was developed. The method was validated by parameters: selectivity, lower limit of quantification, linearity range, accuracy, and precision. The pharmacokinetics was linear in the dose range of 5-25 mg/kg after oral administration. The mean maximal concentration in plasma (Cmax) was dose dependent and it was 37.12-114.71 pg/mL for various doses, area under the curve (AUC0-24) was 192.92-597.14 h • pg/mL, mean time to reach maximum plasma concentration (Tmax) was 3-3.5 h, half-life (T1/2) was 7.88-9.89 h, and the mean retention time (MRT) was 1.73-14.31.Conclusion. After nontraditional approach based on correlation of special marker activity (DPP-4) and active substance concentration the plasma pharmacokinetics of sea urchin gonads extract after oral administration to the rabbits in three doses was characterized. Similar approaches may be effective for compounds and complex mixtures when it is difficult or impossible to analyze them traditionally by chromatographic (HPLC-UV/FL/MS, GC-MS, etc.) methods.

The aim of this work was to study the distribution of the labeled 123I peptide-protein drug Cellex in organs and tissues of rats after single intravenous injection and accumulation. After a single intravenous injection of radiolabeled [123I]Cellex it was found that the drug is characterized by rapid distribution into tissues and organs. The highest content of the drug in all organs and tissues was observed in 15 minutes after its administration. It was established that the drug penetrates the blood-brain barrier and about 6% of the drug accumulates in the brain, which is 10% of the total amount of the drug found in all organs in 15 minutes after administration.

The high demand for insulin preparations by the population of the country makes it urgent to create domestic bioanalogues for the treatment of diabetes mellitus. The equivalency of the quality, safety and efficacy of the bioanalogue with the reference medicinal preparation registered in Russia is required by authority at the stage of preclinical studies. The aim of the study was to study the interaction of insulin substances of domestic production, including short and long-acting insulin with the insulin receptor in comparison with the reference substances registered in Russia by the surface plasmon resonance method. Comparing the interaction kinetics curves and calculated dissociation constants determined by the surface plasmon resonance method it was confirmed that there are no statistically significant differences between the potentially bioanalogical insulin substances Rosinsulin R, Rosinsulin Glargin and Rosinsulin Aspart (LLC ”Zavod Medsintez”, Russia) and the corresponding substances of registered in Russia medicines.

Ethnopharmacological relevance: Aqueous autolysate from the snake Eryx miliaris (SNA) has been used in traditional medicine of Uzbekistan as anti-inflammatory, hepatoprotective and immunomodulatory agent. However, little is known about the chemical composition and its mechanisms of activity. Aim of the study: This is our first attempt to analyse the composition of snake autolysate using gas chromatography with mass spectrometry (GC-MS) and to investigate the mechanisms of anti-inflammatory and hyaluronidase activity of fingerprinted E. miliaris autolysate to support their use in the traditional Uzbek medicine. Materials and methods: Aqueous autolysate was evaporated and derivatised for GC-MS analysis of metabolites. For quantification, lipids were extracted from autolysate by solvent extraction and derivatised by esterification and silylation. Biological activity was evaluated with lipid peroxidation, cyclooxygenase (COX) inhibition and antihyaluronidase activity tests. Results: GC-MS analysis of SNA enabled the identification of 27 compounds. Short chain fatty acids (SCFA, 21%), amino acid/derivatives 39% (incl. 2-piperidinone 19%), phenyl (7%), and OH-Phenyl (10%) derivatives covered 77%. Other derivatives (9%) included succinic acid and 3-indole acetic acid). Long chain fatty acids (C-16-C-18) accounted for 3%. The lipid concentration of SNA was 1.2 mg/mL (0.12%). Three concentration levels (1.0-20.0 mu g/mL) did not inhibit COX-1 and COX-2 in vitro and malondialdehyde level was not decreased by SNA in lipid peroxidation model. However, SNA was a potent inhibitor of the hyaluronidase enzyme activity in a dose dependent manner with IC50 = 0.086 mL/mL. Conclusion: The results from GC-MS analyses of SNA lead us to the identification of a wide range of major chemical structures of the metabolites and their derivatives with several categories. Pharmacological studies support the traditional use of SNA and show one of its possible mechanisms of activity via inhibition of hyaluronidase.

Extraction of lipophilic substances from shells and spines of green sea urchins (Strongylocentrotus droebachiensis) was studied. It was found that the extraction time had the greatest influence on the yield of lipophilic substances whereas temperature and extraction modulus had insignificant effects. Ethanol (95%) was chosen as the extractant affording the maximum amount of lipophilic substances. Optimization of the defatting step established that the maximum yield of lipophilic substances (1.2%) was attained after extraction for 3 h at 55°C with a 1:8 raw-material—extractant ratio. Free fatty acids (57%), phospholipids (~7%), diglycerides, sterols, triglycerides, and hydrocarbons were detected in the lipophilic extract. Phospholipids in the lipophilic extract were represented mainly by phosphatidylcholine. Lysophosphatidylcholine, lysophosphatidylethanolamine, phosphatidylinositol, and phosphatidylethanolamine were detected in trace quantities. The (poly)hydroxynaphthoquinone pigment content after purifying the raw material of lipophilic substances was 98.5% according to HPLC.

Изучен процесс экстракции липофильных веществ из панциря с иглами зеленого морского ежа. Установлено, что наибольшее влияние на выход липофильных веществ оказывает время экстракции, в то время как температура и модуль экстракции оказывают незначительное влияние. В качестве экстрагента, извлекающего максимальное количество липофильных веществ, выбран 95 % этиловый спирт. В результате оптимизации процесса обезжиривания установлено, что максимальный выход липофильных веществ (1,2 %) достижим при обработке сырья в течение 3 ч, при температуре экстракции 55 °C и соотношении сырьё — экстрагент (1:8). В липофильном экстракте обнаружены 57 % свободных жирных кислот, около 7 % фосфолипидов, диглицериды, стерины, триглицериды и углеводороды. Фосфолипиды в липофильном экстракте представлены преимущественно фосфатидилхолином. Лизофосфатидилхолин, лизофосфатидилэтаноламин, фосфатидилинозитол и фосфатидилэтаноламин обнаружены в следовых количествах. Содержание полигидроксинафтохинонового пигмента после очистки сырья от липофильных веществ по данным ВЭЖХ составило 98,5 %.

•Immunomodulatory and antioxidant properties of fixed combination of fish oil with schisandra oil extract (CFS) and chamomilla oil extract (CFM) were evaluated in vivo and in vitro.•The fixed combinations exhibited inhibition of the delayed-type hypersensitivity in mice.•CFM showed stimulation of humoral immunity and increase of secondary antibody titer.•Combination of fish oil with plant extracts lead to significant synergistic antioxidant effect.

During this work it was studied the influence of the new entretenimiento of natural origin (DPP-4 inhibitor, the drug KLS-073) on basic biochemical parameters for a long period (12 months) of application. The experiment was carried out on outbred mice of both sexes. The introduction of the drug was carried out during 12 months in three doses: higher therapeutic (ITD) (2 mg/kg), 5 ITD (10 mg/kg) and 10 ITD (20 mg/kg). As of the evaluation criteria in the assessment we used overall biochemical and hematological parameters and histological examination of the thyroid and the pancreas over time (the 31st, the 91st, 181st and 361 th days). According to the results, the study showed no differences in biochemical parameters and structure of the thyroid and pancreas glands of intact animals and animals treated with the study drug KLS-073.

Fish, like other aquatic organisms, are a potential source of structurally diverse bioactive compounds. Studies of the pharmacological effects of fish peptides have revealed their antihypertensive, immunomodulatory, antioxidant, antitumor, and antimicrobial activities. Analysis of the literature data confirms that fish can be used not only for nutritional purposes, but also as a source of unique peptides with a broad spectrum of biological activities. Further investigations will allow the inclusion of fish peptides as acting agents in modern medicinal drugs.

The regular sea urchin Strongylocentrotus pallidus is a widespread species in high-Arctic waters. However, little is known about lipid composition of sea urchin eggs from northern Barents Sea.

The aim of work was comparison of two detection methods of TLC plates for quantification of phosphatidylcholine (PC) in sea urchins eggs. Lyophilized sea urchins eggs from Barents Sea were used. Lipids fraction was extracted with chloroform/methanol (2/1) by sonification in 30min. A samples were spotted on Silica gel 60 F 254s glass plates (Merck, Germany) using a Linomat V system (Camag, Switzerland). The plates were developed in mixture of chloroform/methanol/acetic acid/water (7/2/0.8/0.5) [1]. PC was quantified by direct densitometric scanning of the developed plate at 202nm and after derivatization with 2% phosphomolybdic acid solution at 700nm using a Camag TLC Scanner 3. The best separation of PC, sterols, fatty acids and triglycerides was obtained. Characteristics of various techniques of PC quantification are resulted in Table 1. The quantitative results of both detection methods did not show any statistically significant differences between each other.

Nanosizing is an advanced formulation approach to address the issues of poor aqueous solubility of active pharmaceutical ingredients. Here we present a procedure to prepare a nanoparticulate formulation with the objective to enhance dissolution kinetics of taxifolin dihydrate, a naturally occurring flavonoid with antioxidant, anti-inflammatory, and hepatoprotective activities. Polyvinylpirrolidone was selected as a carrier and the solid nanodispersions of varying compositions were prepared by a co-precipitation technique followed by lyophilization. The formulation technology reported herein resulted in aggregate-free, spherical particles with the mean size of about 150nm, as observed by scanning electron microscopy and measured by photon correlation spectroscopy. Furthermore, the co-precipitation process caused taxifolin dihydrate to convert into an amorphous form as verified by X-ray powder diffraction, differential scanning calorimetry, hot stage microscopy and Raman spectroscopy. Finally, in vitro dissolution behavior of the nanodispersion of taxifolin was shown to be superior to that of either pure drug or a drug–polymer physical mixture, reaching 90% of taxifolin released after 30min. Such enhanced drug release kinetics from the nanodispersion was attributed to both the reduced particle size and the loss of crystallinity.

Two chromatographic methods, high-performance TLC (HPTLC) and HPLC, were developed and used for separation and quantitative determination of chlorogenic acid in green coffee bean extracts. For HPTLC silica gel Kieselgel 60 F 254 plates with ethyl acetate/dichlormethane/formic acid/acetic acid/water (100:25:10:10:11, v/v/v/v/v) as mobile phase were used. Densitometric determination of chlorogenic acid by HPTLC was performed at 330 nm. A gradient RP HPLC method was carried out at 330 nm. All necessary validation tests for both methods were developed for their comparison. There were no statistically significant differences between HPLC and HPTLC for quantitative determination of chlorogenic acid according to the test of equality of the means.