Introduction. Hypertension is the most common non-infectious disease in the world. New clinical recommendations for the diagnosis and management of patients with arterial hypertension are considering the issue of prescribing combination therapy and prefer fixed combinations of drugs in a single pill. The study of the pharmacokinetics of medicinal substances and the consideration of their pharmacokinetic parameters today is a necessary step in the complex of work, both in the creation of new original medicines and in the application of known generic drugs, and this is primarily due to obtaining objective characteristics of all processes occur in the body of the animal (human) with the drug. Pharmacokinetics is assessed in individual studies or as part of efficacy, safety, and tolerability studies.

Aim. The study of the release of lercanidipine from bilayer tablets containing two API (ramipril and lercanidipine) in the dissolution medium used for quality control in vitro and release in vivo, after oral administration of the drug to rabbits.

Materials and methods. Studies have been conducted on the release of lercanidipine from the combined drug in vitro and in vivo. As a test system were used laboratory rabbits Soviet chinchilla breed. Pharmacokinetic parameters were determined.

Results and discussion. A graph of the release of lercanidipine from the combined drug was constructed and the dependence of the concentration of this substance in the blood plasma of rabbits on time was revealed. Calculated pharmacokinetic parameters. An in vivo release study shows that the pharmacokinetics of lercanidipine are consistent with literature data.

Conclusion. The test drug has all the advantages of a rational fixed combination of antihypertensive drugs and simplifies therapy, meets the requirements of the latest clinical guidelines.

Introduction. Herbal drugs of black walnut (Juglans nigra L.) are not registered on the territory of the Russian Federation at the present time. However, the State Register of Medicinal Products includes an extract of unripe fruits of a related species walnut (Juglans regia L.) «Yuglanex» (made in Russia) and the complex herbal drug «Tonsilgon» (made in Germany). A number of biologically active food supplements from black walnut raw materials have been registered by Russian Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing. They are an additional source of phenolic compounds as tannins and flavonoids. The purpose of the article is to study and systematize up-to-date information on the chemical composition of the medicinal plant raw material of black walnut and the pharmacological properties of its main biologically active compounds.

Text. All parts of the black walnut are used in the ethnomedicine of the indigenous population of North America according to indications similar to the walnut in Asia and the Manchurian nut in the Far East: snake bites, fever and disorders of the gastrointestinal tract.

The chemical composition of the medicinal plant raw material of black walnut is dencently walnut by its qualitative composition. Fruits, bark and leaves of black walnut contain a rich polyphenol complex (naphthoquinones, in particular, juglon and its derivatives, tannins, flavonoids, phenolic acids), vitamins, essential oil, organic acids. However, the quantitative analysis revealed a higher content of biologically active substances in the raw black walnut, especially in relation to polyphenolic compounds.

The scientific literature describes the results of experiments on animals, confirming antioxidant, antimicrobial, antifungal, antiviral, antiparasitic, hypoglycemic, antispasmodic, and anti-tumor effect on certain cell lines.

Conclusion. As a result of studying the literature and systematizing the current information on the chemical composition of the medicinal plant raw material of the black nut and the pharmacological properties of its main biologically active compounds, it has been established that the main properties are related to the presence of the phenol complex. However, a deeper study of the chemical composition is required. Both the total extracts of the black walnut from plant raw materials and individual compounds show predominantly antimicrobial, antifungal, antioxidant, antiviral, hypotensive, immunomodulatory, antitumor and antispasmodic activities in preclinical trials.

Introduction. Nowadays, due to the emergence of new broad-spectrum antibiotics and synthetic chemotherapeutic agents, the use of nitrofuran has significantly decreased. At the same time, they remain important for the treatment of urinary tract infections and intestinal infections, as well as local antiseptics. However, low solubility in water, the possibility of undesirable reactions, as well as the absence of prolonged action significantly limit their use. It is known that the inclusion of biologically active substances in the polysaccharide molecule allows improving the already known drugs while changing their properties. Thus, the introduction of the 5-nitrofuran cycle in native alginic acid and some of its derivatives led to the emergence of antifungal activity and improved solubility. However, the information about the antimicrobial activity of N-arylidene(alkyliden)hydrazides of carboxymethylalginic acid in the literature are practically absent.

Aim. Synthesis and study of antimicrobial activity of N-arylidene(alkyliden)hydrazides of carboxymethylalginic acid with different content of a biologically active fragment.

Materials and methods. For the proof of the structure of the synthesized substances were used IR- and UV-spectroscopy, antimicrobial activity was analyzed by double serial dilution on the test-cultures Staphylococcus aureus, Escherichia coli and Candida albicans.

Results and discussion. With the introduction of 5-nitrofurfural and β-(5-nitro-2-furyl)-acrolein into the carboxymethylalginic acid, samples with antimicrobial activity were obtained, whose spectrum of action differs from furacilin. Thus, in the polymer acylhydrazones β-(5-nitro-2-furyl)-acrolein of alginic acid, a high antifungal action against C. albicans was found, the fungistatic and fungicidal action reaches 1,8 and 3,6 µg/ml.

Conclusion. The synthesized samples of polymer acylhydrazones 5-nitrofurfural and β-(5-nitro-2-furyl)-acrolein have antimicrobial action, and unlike furacilin are highly soluble in water and do not irritate the skin and mucous membranes. The content of 5-nitrofuran fragment, at which the maximum antimicrobial activity is achieved, is revealed.

Introduction 

Introduction. According to the literature, modern technologies, in particular, extraction in an ultrasonic field, make it possible to obtain concentrates of biologically active substances (BAS) with almost complete preservation of the chemical composition inherent in natural raw materials and high yield of extractive substances. At the same time, the ability to regulate the concentration of recoverable active substances during the technological process opens up prospects for the use of natural components as the main pharmaceutical substance.

Aim. The purpose of this study was to select the optimal extraction method for obtaining extracts with a high content of BAS from fenugreek seeds. 

Materials and methods. Extracts were obtained at a temperature of 60±2 °С, by the method of dynamic maceration using an ultrasonic device. Ultrasonic extraction was performed using an ultrasonic installation I100-6/4. In both cases, purified water and ethanol solutions in various concentrations from 40 to 90% were used as extractants. The ratio of raw materials: extractant was as 1:10 (by weight). A portion of vegetable raw materials (10 g) was placed in a glass and poured 100 cm³ of extractant. Next, maceration or sonication was performed. Maceration was performed with magnetic stirring, the rotation speed was 100 rpm. Extraction was carried out at a temperature of 55–60 °C for 5 hours. Ultrasonic effects on solid plant materials were carried out with an intensity in the range from 17 to 22 kHz for 30–60 minutes.

Results and discussion. Studies have allowed to determine the amount of extractives in the seeds of fenugreek hay, and choose the most promising method of extraction to obtain extracts with a high content of biologically active substances. A comparison was made between dynamic maceration and ultrasonic extraction. The most promising was the method of ultrasonic extraction at an oscillation frequency of 22 kHz. For 1 hour of extraction, it was possible to achieve depletion of raw materials. It is established that the content of extractable substances directly proportional depends on the duration of treatment and the frequency of ultrasonic waves. The correlation coefficient was 0.78.

Conclusion. Ultrasonic extraction when exposed for 60 minutes allows you to get a greater amount of extractives compared with dynamic maceration. To achieve the same level of extractive substances by the method of dynamic maceration, the raw materials must be processed within 5 hours.

Introduction. The article presents data on the development and implementation of a method for measuring the moisture content of bulk material during its pneumatic transport in pharmaceutical production. The method of measuring moisture is theoretically based on the physical Pockels effect. An analysis of the currently existing pharmacopoeial methods for monitoring the quantitative water content and methods of in-process control of the drying process is presented. The development of new methods and technical means that provide the necessary speed and accuracy of moisture measurement is an important task, especially in the pharmaceutical industry to control the residual moisture of the starting materials, intermediate products during their production and finished products.

Text. Analysis of the theoretical assumptions for the method and technical systems for measuring the mass of bulk dielectric materials during their pneumatic conveying shows the ability to measure the moisture content of the bulk dielectric material transported by air through a pipeline in a new non-contact manner. The proposed method is based on one measurable parameter – the intensity of the light wave passing through the Pockels cell. A block diagram of the system developed by the authors for automated moisture measurement of bulk dielectric materials with an electrooptical Poсkels cell is presented. The article describes a method for measuring moisture, based on the Pockels effect, for bulk material during its pneumatic transport.

Conclusion. The proposed method of measuring moisture is contactless, just technically feasible, has a high speed and measurement accuracy, since it is based on the Pockels effect. The method is universal for determining the moisture content of bulk dielectric materials in the process of their pneumatic transportation. The above advantages of this method suggest the possibility of its successful application in pharmaceutical production.

Introduction. The development of the composition of drugs is very difficult and long process. To implement it, it is currently recommended to apply a modern approach – quality through the development of Quality by design. One of the methods for introducing this approach is the use of multifactorial experiments.

Aim. The aim of the work was to develop the composition and technology of effervescent tablets using the Minitab program.

Materials and methods. For the study used the program Minitab (ver. 18.), Minitab LLC, USA. The program created an experiment plan and evaluated those obtained on the basis of its models using regression analysis.

Results and discussion. The composition and technology of effervescent tablets was developed by direct compression using the principles of Quality by design. Algorithms of actions for using the Minitab software package for development are proposed. An experiment plan was constructed and models of the dependence of quality indicators on the composition and technological parameters were generated. The resulting sub-sections were analyzed using regression analysis tools.

Conclusion. Based on the results of the work, the composition of effervescent tablets obtained by direct compression technology, as well as the algorithm of drug development in the Minitab program, was proposed.

Introduction. Today, innovative technologies are widely used in pharmacology (in particular, in the production of herbal preparations), in cosmetology (obtaining various extracts and oils, complex preparations), as well as in the food industry (as natural dyes, etc.). Plant extracts with a high level of risk do not have a harmful effect on the human body, except that they provide environmental safety, which creates a special interest for the pharmaceutical industry . vibrocavitation and supercritical fluid СО₂ extraction.

Aim. The choice of the optimal level of extraction to obtain a high level of fenugreek seeds.

Materials and methods. Obtaining a vibrocavitation extract with an experimental vibration-explosive installation made at the Department of Processes and Apparatus of the St. Petersburg State Technology Institute. Extracts at a temperature of 60±2 ºС. The frequency of revolutions of the homogenizer ranged from 1000 to 5000 rpm. Ultrasonic impact using ultrasound unit I100-6/4 Ultrasonic effect on solid plant material with an intensity of 22 kHz for 60 minutes. The time of extraction in a vibro-cavitation extractor-homogenizer was studied for optimal values.

Supercritical fluid СО₂-extraction is obtained in two versions with the use of extragent (96% ethanol in the presence of carbon dioxide: ethanol 9:1) and without it. Extraction of the sound of a supercritical fluid extraction system with a 1-liter vessel. SFE1000-2-BASE with a kit for upgrading the SFE1000-2-BASE system to an SFE1000M1-2-FMC50 system (Waters, USA). The flow rate of the extractant was 60 g/min. Emergency listening for one hour and pressure 200, 300 and 400 bar. Extraction is observed three times. The obtained extracts indicate the amount of extractive substances according to article of Russian state pharmacopoeia 1.5.3.0006.15 «Determination of the content of extractive substances in plant raw materials and medicinal plant preparations». Quantitative determination of the saponin complex of parasitic seed seeds on an Agilent QTOF-6530 chromatograph with two ESI and APCI ionization sources according to Gravel et al.

Results and discussion. Studies have allowed to determine the amount of extractives in the seeds and choose the most promising method. 

Conclusion. As a result of our research, it was found that the most promising extraction method for extracting seeds is a parasitic with vibration extraction at frequent revolutions of the homogenizer of 5000 rpm and an extraction time of 60 minutes.

Introduction. Antiseptic Dorogov's stimulator 3^rd fraction (ASD-3) is a multicomponent substance, which is obtained by the thermal processing of cattle-origin tissues, which contains more than 120 substances, including biologically active substances. Among the components of ASD-3, necessary for quantitative analysis, pyrrole, phenol, cresol and its derivatives, and indole are distinguished as components. Despite the fact that antiseptic Dorogov's stimulator 3^rd fraction was discovered more than half a century ago, there is no information about the pharmacokinetics of the drug components to date. Phenol is one of the preferred substances for research because that is a substance applied in the pharmaceutical technology for the manufacture of medicines.

Aim. Study of pharmacokinetic parameters of phenol as a component of a drug containing an antiseptic Dorogov's stimulator 3^rd fraction in a dose of 0.5 g/kg, with a single use in a blood serum of 84 female rats weighing 100–120 g.

Materials and methods. In the blood serum of the 84 female Wistar rats weighing 100–120 g a study of a phenol pharmacokinetics as a component of ASD-3 drug in a dose of 0.5 g/kg was performed by a single skin application. The first group of animals was intraperitoneally injected with an aqueous emulsion containing 5% ASD-3, while the second one received zinc paste containing 5% ASD-3 on a skin surface. After the application of the emulsion or paste, the animals were euthanized after 0.25; 0.5; 1; 1.5; 2; 6 and 8 hours with further collection of blood from the abdominal aorta into centrifuge tubes. The study was carried out by high performance liquid chromatography.

Results and discussion. The elimination rate constant for intraperitoneal drug emulsion injection is 0,0968 h^-1, and when the drug paste is applied, 0.0581 h^-1. As a result, the half-life of phenol in the first case is 7.2 hours, and in the second case – 11.9 hours. The relative bioavailability of the phenol from the paste containing 5% of the ASD-3 is 14.38% to the 5% emulsion injected into the rats body intraperitoneally.

Conclusion. Comparison of the phenol concentration in the blood serum of experimental animals in two ways of application showed that the skin application of a drug paste containing 5% of ASD-3 provides a 10 times lower content of phenol in the blood serum compared to the intraperitoneal application of an emulsion with 5% ASD-3. 15 minutes after the skin application, the components of the drug start to be detected in the blood serum, reaching its maximum concentration in 1.5–2 hours, subsequently gradually decreasing.

Introduction. Even though a wide range of the products for caries treatment is available, the efficiency of its treatment with the traditional methods is 62,5–75,4%. As a result of the set of researches carried out at the Department of Pharmaceutical Technology of the Perm State Pharmaceutical Academy, in cooperation with the Department of Dentistry of the Advanced Training and Professional Retraining Faculty of E. A. Wagner Perm State Medical University, a medical method of moderate and advanced caries treatment with remineralizing gel and films is proposed. During the preparation of the product specification file for the dentin caries treatment film, validation of the methods of the active ingredients quality control has been undertaken.

Aim. Validation of the methods of identity tests and of potency assays of dentin caries treatment films.

Materials and methods. In order to achieve the aim, active pharmaceutical substances of pharmacopoeial quality have been used. As a base for developing the methods of identity tests the methods with distinctive reactions to cations and anions have been taken. For the potency assay of calcium chloride a complexometric back-titration has been used; photoelectric colorimetry for potassium phosphate dibasic and sodium fluoride; spectrophotometry for chlorhexidine bigluconate. Five production samples of the films were the object of the research.

Results and discussion. Basing on the research results, the choice was made in favour of the methods of identity tests that are characterized by negative analytical response when applied to the model mixtures which are free from analyte and placebo, and by positive analytical response when applied to the model mixtures of various composition containing analyte. The validation characteristics of the methods of dentin caries treatment films potency assays, which may be included in the product specification file, have been studied.

Conclusion. Validation results show that the chosen methods are specific for both identity tests and potency assays of active substances in the dentin caries treatment films; they are characterized by precision, repeatability and linear dependency in the analytical range of ±30% in regard to the specified active substance concentration in the films. All that allows using them for a reliable quality assessment.

Introduction. IImmunogenicity identification of therapeutic proteins, such as human insulin analogues, is one of the most relevant and significant area in medicine and pharmaceuticals. Determination the possibility of producing neutralizing antibodies to insulin reducing the therapeutic effect of the drug, is an important step to understand the pharmacological profile of the drug. Applying of cell-based methods one allows to determinate neutralizing antibodies to insulin.

Aim. Development and validation methods for detection of neutralizing antibodies against insulin in human plasma.

Materials and methods. The method is based on the use of the iLiteTM Insulin Assay Ready Cells [1], in the genome of which the firefly luciferase reporter gene is introduced under the control of an insulin-dependent promoter. As the insulin concentration increases, the firefly luciferase expression (Firefly) increases, allowing one to use this cell line to estimate the number of neutralizing antibodies against insulin. For normalization by the number of cells and considering the matrix effect of studied samples, the second reporter gene luciferase Renilla is used, which is expressed under the control of a constitutive promoter. The activity of both luciferases was measured using the DualGlo Luciferase Assay System (Promega) assay [2].

Results and discussion. Optimal insulin concentration and plasma/serum dilution were determined to identify neutralizing antibodies to insulin. The long-term stability of neutralizing antibodies to insulin were shown in human plasma for more than 3 months. The developed method was applied in a comparative research of the safety and immunogenicity of insulin analogues (Glargine). Method for the determination of antibodies to insulin was.

Conclusion. A method for determination of neutralizing antibodies to insulin in human K₂EDTA plasma was developed and validated using iLiteTM Insulin Assay Ready Cells system; based on the binding of the insulin alpha chain to the high-affinity heterodimeric CD220 receptor.

Introduction. Busereline, being a synthetic gonadotropin-releasing hormone analog, is widely used for hormone-dependent cancer treatment (e.g. prostate cancer and breast cancer). Based on the accumulated scientific data for busereline quantitation in biosamples, the main analytical method that is used for this purpose is high-performance liquid chromatography (HPLC) with fluorescence detection, combined with protein precipitation (TCA 10%) for sample preparation. However, due to several limitations of this method resulting in low sensitivity (at the µg/mL level of concentrations), the HPLC-MS/MS analytical method was chosen for peptide determination in biosamples. The HPLC-MS/MS method is considered to have higher accuracy and specificity. The main sample preparation method for gonadotropin-releasing hormone analogs is solid-phase extraction. In our work, we’ve chosen protein precipitation as an alternative – easier and less laborious biosamples preparation process.

Aim. The main objective of this study was the development and validation of HPLC-MS/MS method for busereline quantitation in animal (mini pigs) plasma samples and its further application to pharmacokinetic studies.

Materials and methods. Busereline quantitative determination in plasma samples was performed using HPLC-MS/MS method. A protein precipitation procedure (methanol, 1:2, v/v) was used for busereline extraction from pig plasma.

Results and discussion. The developed analytical method was validated for selectivity, linearity, matrix effect, accuracy (intra-day, inter-day), precision (intra-day, inter-day), LLOQ, carryover and stability.

Conclusion. A new HPLC-MS/MS method for busereline quantitation in blood plasma was developed and successfully validated. The developed method showed linearity over the quantitation range from 1 to 20 ng/mL. The developed method can be successfully applied to busereline pharmacokinetic studies.

Introduction. The article is devoted to the relevance of choosing sample preparation method for heavy metals concentration determination in biological sample by using atomic emission spectrometry. Much attention is given to compare 2 methods of sample preparation: solid-phase extraction and microwave decomposition. As the result of analysis of heavy metals Fe, Zn, Rb, Cu, Ni, Al, Mn content using these sample preparation methods the following conclusions are drawn: maximum extraction of heavy metals is achieved by using microwave decomposition method.

Aim. The purpose of this work was to compare the methods and conditions for the sample preparation of biological samples for the determination of the content of heavy metals using atomic emission spectrometry.

Materials and methods. The object of the study to determine the content of heavy metals (Zn, Rb, Cu, Ni, Al, Mn) was the liver of golden hamsters of both sexes weighing 60–145 g of the type Golden Syrian SPF category, obtained from the Federal Research Center Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences (Novosibirsk). Quantitative determination of heavy metals in biological samples was carried out on an Agilent 4100 atomic emission spectrometer with a microwave plasma (Agilent Technologies, USA).

Results and discussion. Comparison of the content of elements in the sample using the sample preparations under study showed that the maximum extraction of heavy metals from biological samples is achieved when using the method of microwave decomposition. The most complete extraction of heavy metals from biological samples is achieved by extraction within 30 minutes.

Conclusion. This method warrants the maximum metal extraction, is more accurate, rapid and less labor intensive compared to other methods considered, and is suitable for determining most heavy metals.

Introduction. Bevacizumab is a monoclonal IgG1 antibody that binds to and inhibits the biologic activity of human vascular endothelial growth factor (VEGF). Bevacizumab is used as a targeted monoor combination therapy for different solid tumors. Phase I clinical trial was performed to assess pharmacokinetics (PK) and immunogenicity of bevacizumab drugs. For this study 80 healthy male volunteers were recruited and randomized to either Avastin or RPH-001 group.

Aim. To assess and compare pharmacokinetics and immunogenicity (safety) following single intravenous administration of Avastin® (manufactured by F. Hoffmann-La Roche Ltd., Switzerland) and bevacizumab biosimilar RPH-001 (manufactured by R-Pharm Group, Russia).

Materials and methods. Bevacizumab quantitation and quasi-quantitative anti-bevacizumab antibodies detection in human blood serum were carried out using photometric ELISA. Two different methods were successfully validated.

Results and discussion. Bevacizumab quantitation method was validated for selectivity and specificity, calibration curve, sensitivity, accuracy and precision, minimal required dilution, dilution linearity and stability. The anti-bevacizumab antibodies detection method was validated for cut-point (with normalization factor calculation), selectivity, sensitivity, precision, drug tolerance, dilution linearity, matrix effect (in case of serum hemolysis), and stability. The validated methods were successfully applied to pharmacokinetic and immunogenicity assessment of bevacizumab drugs.

Conclusion. The results of the PK-study showed that test and reference bevacizumab drugs were equivalent. Immunogenicity study did not show any evidence of anti-bevacizumab antibodies in blood serum samples.

Introduction. Telmisartan is a modern antihypertensive drug, from the group of angiotensin II receptor antagonists, which, recently, has been the purpose of the development and registration of generic drug. The pharmaceutical development of telmisartan generics and their evaluation in bioequivalence studies is complicated by many factors that influence the results of comparison with the reference drug. Therefore, it is relevant to study these factors for the proper planning of studies and for evaluating their results from the generic drugs of telmisartan.

Aim. The aim of the study is to identify the key factors that should be considered in planning and evaluation of bioequivalence studies of telmisartan’s generics.

Materials and methods. To identify the key factors influencing the planning and evaluation of studies of telmisartan generics, a retrospective analysis of the results of seven telmisartan studies was conducted. Calculated pharmacokinetic parameters C_max, AUC_0-t, t_max; their average values; the values of intraindividual variability for each study and the average of the coefficients of intraindividual variability obtained.

Results and discussion. The results of the study demonstrated that the drugs of telmisartan are highly variable, there are sex differences in the pharmacokinetics of telmisartan. According to the averaged values of the concentrations and pharmacokinetic profiles of telmisartan, the following areas were determined: the area of maximum exposure of telmisartan, the duration and schedule of blood sampling, to obtain the optimal pharmacokinetic profile. The most optimal method for the determination of telmisartan is HPLC MS/MS with a lower limit of quantification in the range of 0.5–3 ng/ml. The upper limit of the 90% confidence interval was calculated for the pooled average of the coefficients of intraindividual variability obtained in the considered studies.

Conclusion. Recommendations were developed for planning and evaluating bioequivalence studies of generic telmisartan preparations. It is necessary to plan bioequivalence studies with replicated design in three or four periods, with two crossovers, with two sequences of drug intake. To assess the pharmacokinetics of telmisartan one should use the data on its exposure given in the article. When determining the sample size, one should be guided by the upper limit of the confidence interval of the averaged values of the coefficient of intra-individual variability C_max  of telmisartan (45%). Evaluation of results should be carried out taking into account the possibility of scaling bioequivalence limits for the C_max. The bioequivalence limit for the AUC_0-t  parameter should remain in the range of 80.00–125.00%, as well as the ratio of geometric means for both parameters.