Метилювання положення 8 піридинової частини молекули N-(бензил)-2-гідрокси-4-оксо-4Н-піридо[1,2-a]піримідин-3-карбоксамідів як спроба посилення їх аналгетичних властивостей

The chemical modification of the pyridine moiety of the molecule – displacement of the methyl group in position 8 of pyrido[1,2-a]pyrimidine nucleus has been considered as one of the possible versions to optimize the biological properties of N-(benzyl)-2-hydroxy-4-oxo-4H-pyrido[1,2-a]pyrimidine-3-carboxamides. The synthesis of the research targets was carried out by the reaction of the corresponding benzylamines and ethyl 2-hydroxy-8-methyl-4-oxo-4H-pyrido[1,2-a] pyrimidine-3-carboxylate, in its turn obtained by condensation of 2-amino-4-methylpyridine (i.e. the product with the methyl group in the intentionally required position) and triethyl methanetricarboxylate. The structure of the compounds obtained has been confirmed by the data of elemental analysis and NMR 1H spectroscopy, and in the case of optically active 1-phenylethylamides additionally by polarimetry. The study of the analgesic properties of all N-(benzyl)-2-hydroxy-8-methyl-4-oxo-4H-pyrido[1,2-a]pyrimidine-3-carboxamides was performed on the standard experimental “acetic acid writhing” model. At the same time, it has been found that our modification is accompanied with the increased biological activity of exclusively para-substituted derivatives. For profound research 4-fluorobenzylamide exceeding Piroxicam and Nabumetone by the level of the specific effect has been recommended as a potential new analgesic.В качестве одного из возможных вариантов оптимизации биологических свойств N-(бензил)-2-гидрокси-4-оксо-4Н-пиридо[1,2-a]пиримидин-3-карбоксамидов рассмотрена химическая модификация пиридиновой части их молекулы – перемещение метильной группы в положение 8 пиридо[1,2-a]пиримидинового ядра. Синтез целевых объектов исследования осуществлен реакцией соответствующих бензиламинов с этил-2-гидрокси-8-метил-4-оксо-4Н-пиридо[1,2-a]пиримидин-3-карбоксилатом, в свою очередь, полученным конденсацией 2-амино-4-метилпиридина (т. е. продукта с метильной группой в заведомо требуемом положении) с триэтилметантрикарбоксилатом. Строение синтезированных веществ подтверждено данными элементного анализа и спектроскопии 1Н ЯМР, а в случае оптически активных 1-фенилэтиламидов дополнительно еще и поляриметрически. Изучение анальгетических свойств всех N-(бензил)-2-гидрокси-8-метил-4-оксо-4Н-пиридо[1,2-a]пиримидин-3-карбоксамидов проведено на стандартной экспериментальной модели уксуснокислых корчей. При этом найдено, что предпринятая нами модификация сопровождается усилением биологической активности исключительно паразамещенных производных. Для углубленных испытаний в качестве нового потенциального анальгетика рекомендован 4-фторбензиламид, превосходящий по уровню специфического эффекта Пироксикам и Набуметон.Як один з можливих варіантів оптимізації біологічних властивостей N-(бензил)-2-гідрокси-4-оксо-4Н-піридо[1,2-a]піримідин-3-карбоксамідів розглянуто хімічну модифікацію піридинової частини їх молекули – переміщення метильної групи у положення 8 піридо[1,2-a]піримідинового ядра. Синтез цільових об’єктів дослідження здійснено реакцією відповідних бензиламінів з етил-2-гідрокси-8-метил-4-оксо-4Н-піридо[1,2-a]піримідин-3-карбоксилатом, у свою чергу, одержаним конденсацією 2-аміно-4-метилпіридину (тобто продукту з метильною групою в завідомо необхідному положенні) з триетилметантрикарбоксилатом. Будову синтезованих речовин підтверджено даними елементного аналізу та спектроскопії 1Н ЯМР, а у випадку оптично активних 1-фенілетиламідів додатково ще й поляриметрично. Вивчення аналгетичних властивостей усіх N-(бензил)-2-гідрокси-8-метил-4-оксо-4Н-піридо[1,2-a]піримідин-3-карбоксамідів проведено на стандартній експериментальній моделі оцтовокислих корчів. При цьому знайдено, що здійснена нами модифікація супроводжується посиленням біологічної активності виключно паразаміщених похідних. Для поглиблених випробовувань як новий потенційний аналгетик рекомен

Molecular Genetic Studies of Intraoperative Ligamentum Flavum Bioptates of Patients with Spinal Canal Stenosis

Background. Ligamentum flavum is involved in the development of stenosis of the spinal canal and dural sac, but the mechanisms underlying degenerative-dystrophic changes have not yet been sufficiently studied. Aim. To study the expression of candidate genes potentially involved in connective tissue metabolism in intraoperative samples of Ligamentum flavum of patients with stenosis processes of spinal canal and duralsac on the lumbar spine.Materials and methods. During the surgical treatment, Ligamentum flavum biopsies were collected from 33 people with stenosis processes of spinal canal and dural sac on the lumbar spine (16 men, 17 women). RNA was isolated, then reverse transcription was performed. After that, real time PCR was performed (CFX96, Biorad) with the specific primers (RealTimePrimers.com).Results. The characteristics of the expression of candidate genes that are active in the Ligamentum flavum of  the examined patient population are given: NAT1 and NAT2 acetyltransferase genes; genes that determine the intensity of local metabolism (Dio1, Dio2 and Dio3); estradiol receptor genes ESR1 and ESR2; genes encoding receptors for growth factors and parathyroid hormone. All Ligamentum flavum samples showed activity of the NAT, ESR 2, FGFR1, FGFR3, NA20, PTH1R, and PTH2R genes. In the vast majority of samples (93.9–97.0 %), the following genes were active: PDGFA, ESR1, CALCR, and PDGFB. In 21.2–39.4 % of samples, “silence” of the Dio1, Dio2, Dio3, NAT2 and GDF5 genes was detected, and only 39.4 % of samples revealed NAT1 transcripts. The highest heterogeneity of transcript content was observed for Dio1, NAT2, and Dio2. The most stable expression in Ligamentum flavum tissue and a narrow range of fluctuations were characterized by genes with a high level of activity AANAT, ESR2, NAA20 and genes with an average level of activity FGFR1, FGFR3, PTH1R and PTH2R.Conclusions. Ligamentum flavum is a promising little-studied substrate for molecular genetic research. The revealed features of gene expression provide new information about pathogenesis, and the new fundamental knowledge obtained can provide a basis for developing methods for preventing the development of epidural fibrosis

SOME ASPECTS OF THE ESTIMATION OF STRUCTURAL-FUNCTIONAL CHANGES IN MEMBRANE OF ERYTHROCYTES AT THE CARDIOVASCULAR PATHOLOGY

The article presents the estimation of structural-functional changes in erythrocytic membranes accompanied with the change of activity of different enzymes that determine normal functioning of erythrocytes. Change of lipid-protein and protein-protein interactions causes the reapportionment of charges on the surface of erythrocytes and. the decrease of total charge of cell and. leads to the increase of aggregation of erythrocytes and. the change of rheological properties of blood. In the end. it reflects on the effectiveness of oxygen transportation, transcapillary diffusion and perfusion of blood in microcirculation channel

MORPHOLOGICAL CHANGES IN ORGANS AND TISSUES AT SUBCHRONIC INTOXICATION WITH BENZODIAZEPINE MEDICATIONS (EXPERIMENTAL RESEARCHES)

Morphological changes in tissues and organs at subchronic intoxication of white rats with 1,4-benzodiazepines synthetic medications (alprazolame, mezapame, diazepame, and nozepame) are studied in an experiment. Pathomorphological changes in tissues of liver, kidneys, brain and reproductive system that can lead to the development of industrially caused diseases in the employees of benzodiazepines production are revealed

ФАРМАКОЭКОНОМИЧЕСКИЕ ПРЕИМУЩЕСТВА СТРАТЕГИИ ОБЩЕЙ ПРОФИЛАКТИКИ ЦИТОМЕГАЛОВИРУСНОЙ ИНФЕКЦИИ У ПАЦИЕНТОВ ПОСЛЕ ТРАНСПЛАНТАЦИИ ПОЧКИ

CMV infection is a major problem in kidney transplantation patients. Valcyte, Cimevene are the first line thera- py of CMV infection. This pharmacoeconomic analysis shown economic benefits of Valcyte prophylaxis tactic compare preemptive strategy. Развитие цитомегаловирусной инфекции у пациентов после трансплантации почки является серьезной клинической проблемой. Препаратами первой линии фармакотерапии при данном заболевании являют- ся препараты ганцикловир и валганцикловир, однако клинико-экономические аспекты их применения в России изучены недостаточно. Проведенный фармакоэкономический анализ продемонстрировал эконо- мическую обоснованность проведения общей профилактики ЦМВ-инфекции препаратом валганцикло- вир по сравнению со стратегией превентивной терапии.

Modern state of irrigated soils at the south of the Volga upland

The goal is to assess the impact of 50-year irrigation by sprinkling on soil processes occurring in the light-chestnut soils (Luvic Kastanozem (Loamic, Aric, Protosodic, Bathysalic)) of the southern slopes of the Volga upland at the Volga-Don interfluve (FSUE “Oroshaemoe”, the Volgograd region) with deep ground water. Water for irrigation is supplied from the Varvarovsky reservoir of the Volga-Don Canal system. It is characterized by a total dissolved salts of about 1 g/l, a bicarbonate-chloride-sulfate compositionwith an increased sodium content. Detailed morphological description of soil profiles, granulometric composition, content of soluble salts in soils and sediments of the vadoze zone up to the depth of 3.5 m, dynamics of salts in the layer of 0-50 cm for 2011-2019 are presented. Until the autumn of 2015, the studied soils were deep saline, being no saline in the layer of 0-100 cm. In recent years, a weak salinity degree of soda-chloride sodium chemistry has been observed in the 0-50 cm layer as a result of gradual accumulation of irrigation water salts during irrigation organized according to water consumption of agricultural crops. Irrigated soils have acquired a complex of signs of secondary salinity: (1) the presence of light accumulations of sandy and silt mineral grains in the arable horizon, resulting from the destructive effect of irrigation water drops during sprinkling; (2) toxic alkalinity associated with sodium (residual sodium carbonate), according to water extraction 1 : 5 (soil : water), in the horizons from the depth of 10-20 to 60100 cm; (3) abundant humus-clay cutans on the lateral side faces of prismatic structural units in the undisturbed part of the soil profile from 30 to 100 cm

Risk assessment of the adverse impact of industrial synthesis of benzodiazepine drugs on human health

Among the occupational and work-related diseases of employees in pharmaceutical industry the leading positions belong to the diseases caused by long-term work under the conditions of relatively low concentrations of hazard substances. The study and evaluation of risks will eliminate or reduce the impact, thereby, enhancing the quality of life. Synthetic drugs - benzodiazepine derivatives by the values of middle-lethal doses are substances of the 111 class of hazard and cause slight irritating effect to the skin and mucous membranes of the eyes; skin-resorptive and sensitizing effects have not been identified. For the studied benzodiazepine derivatives the safe exposure levels have been approved in the following values: nozepam -1 mg/m3; mezapam - 0.3 mg/m3; sibazon - 0.2 mg/m3; alprazolam - 0.1 mg/m3. On the base of the toxicological studies the parameters of the hazardous effects on the body have been identified allowing for a final risk characterization. The article presents the results of the calculation of the potential dose that may be received by the employee during the working shift in the synthesis of products of this series. Calculation of safe level of exposure is based on a threshold concentration of substances. Drags risk factors by inhalation have been calculated by comparing the value of the potential dose of a xenobiotic received for one shift by inhalation with its level of safe effect under the same route of exposure. The estimated hazard ratios for the studied substances demonstrate a low probability of adverse effects on the workers during the production of nozepam and alprazolam. High risk of negative impact on the health of workers has been identified during the production of sibazon and mezapam. This information can then be used to control the working environment during the synthesis of drugs - benzodiazepine derivatives, aimed at creating a safe working environment and, consequently, improving the workers' life quality

Assessment of the State of Cell Membranes against the Background of Prolonged Use of Anticytokine Therapy in Patients with Ulcerative Colitis

Biological therapy in the treatment of immune-mediated conditions has changed their course, the quality of life of patients and the prognosis of diseases. The accumulated by mankind 20 years of experience with the use of genetically engineered drugs has led to a number of questions regarding, among other issues, safety in the long-term administration of biological therapy. Patients suffering from ulcerative colitis revealed changes in cell membranes, reflecting their structural and energy characteristics. Long-term administration of Infliximab leads to the stabilization of energy processes in the erythrocyte membrane and improves homeostatic function of the kidneys.The aim of the study was to evaluate the effect of long-term use of TNF-α blockers (Infliximab) on the structural and functional characteristics of cell membranes and the functional state of the kidneys in patients with moderate to severe ulcerative colitis.Materials and methods. We examined 103 patients with moderate to severe ulcerative colitis during the period of acute attack and remission, of which 28 patients received basic therapy using the drug Infliximab (IFX) for 10 years, 75 patients received standard basic treatment. The patients of the biological therapy group took the original drug Infliximab – Remicade. The comparison group consisted of 30 healthy volunteers, comparable by sex and age. The analysis of the state of erythrocyte membranes was carried out using a set of physicochemical methods: UV spectroscopy (SF-46m spectrophotometer), high-performance thin-layer reaction paper chromatography, membrane ultrafiltration, erythrocyte NMR spectroscopy on phosphoric (31P) and proton (1H) nuclei. The functional state of the kidneys was evaluated using a dynamic scintigraphic study (with the technemage –Tc-99m).Results. Prolonged use of anticytokine therapy with Infliximab for 10 years in patients with ulcerative colitis, upon reaching deep remission, improves endogenous intoxication, restores the structural and functional characteristics of cell membranes, normalizes cell energy metabolism and does not negatively affect the functional state of the kidneys

Interaction of erythrocyte membranes with derivatives of 3-oxypiridines

In connection with the increased interest in the use of auto-erythrocytes for directional drug transport, the study of the ability of erythrocytes to sorb an antioxidant drug is becoming topical. The article considers the method of spectrophotometry developed for evaluating the ability of erythrocytes to load with a mexidol preparation under conditions of natural sorption and a method created for determining the drug concentration in erythrocyte biological medias. Materials and methods. The peripheral blood of 15 clinically healthy males aged 20 to 45 years was used as an object of the study. A pharmaceutical preparation of oxymethylethylpyridine succinate-mexidol (ZAO «Pharmasoft», Russia) was used for clinical studies. The drug belongs to the group of 3-hydroxypyridines. The inclusion of mexidol was carried out by direct incubation of erythrocytes in a medium containing this preparation. The concentrations of the preparation were 1.25,2.5 and 5 ßg/ml and the incubation time was 15,20 and 3o minutes. The supernatant was obtained by centrifugation for 10 minutes at 3000 rpm on a SF-2000 spectrophotometer at a wavelength of 630 nm. In addition to the pharmacopoeial method, the oxidation-reduction reaction of the supernatant with methylene blue was used. Results. When measuring the spectra of the supernatant with different concentration of the preparation and using methylene blue, a regression relationship between mexidol and optical density was established, and the optimal exposure time of red blood cells and the drug was determined. These data can be used to control the directional transport of the drug to target organs. A model of the equation for determination of the mexidol content in biomedids is proposed