Active Pharmaceutical Substances Research Articles

Production sustained release dosage forms from deliquescent substances by solid dispersion systems

Introduction. The production of solid dosage forms from hygroscopic substances is a complex technological task. The addition of hygroscopic substances into a polymer matrix allows to reduce their moisture absorption capacity.Aim. Reducing the hygroscopicity of deliquescent substances and obtaining sustained release solid dosage forms by using solid dispersion systems.Materials and methods. Substance: DEAE; excipients: Kollidon® VA 64, Soluplus®, PEG 1500, PEG 6000, PEG 8000, Poloxamer Kolliphor® P 188 and Kolliphor® P 407; reagents: hydrochloric acid, sodium dihydrogen phosphate, potassium hydrogen phosphate. The melt of the components was obtained on a twin-screw laboratory extruder and cast into silicone molds. The resulting solid dosage forms were studied for disintegration in three environments corresponding to the sections of the human gastrointestinal tract.Results and discussion. The active pharmaceutical substance DEAE is characterized as deliquescent substance. The introduction of DEAE into PEG-polymer matrices made it possible to reduce the hygroscopicity of solid dosage forms in the form of bars. The introduction of solubilizers into the melt made it possible to obtain bars with the appropriate organoleptic properties. The release of DEAE from bear- or toroid-shaped bars occurs on average 29–47 % faster than from heart-shaped bars.Conclusion. Compositions based on Soluplus® and Kolliphor® P 407 have proven promising for further development of tiles containing DEAE. In order to create tiles with a slow release, it is recommended to use heart-shaped forms, as they have a smaller surface area.

COMPARATIVE ANALYSIS OF METHODS FOR DETERMINING THE CONTENT OF THE TOTAL PROTEIN SARS-COV-2 OF THE COVID-19 VIRUS

One of the main requirements for the new vaccines being created is their safety in use, including the total protein content, which are one of the main indicators of the quality and safety of vaccines. This article presents the results of work on the comparative analysis of methods for determining the total protein in the active pharmaceutical substance (AFS) and also the bulk product of the vaccine "QazCovid-in ® .Determination of the total protein concentration was carried out by comparing the methods of Lowry, Bradford, commercial sets of Clini Test-BL and BSA Gold Protein. As a result of the determination of the total protein in the composition of the AFS by the classical Lowry method and the commercial CliniTest-BL kit, the highest values were obtained on average 616.67 and 589 μl/ml, and in the bulk product 51.11 and 47.38 μl/ml, respectively. The most minimal indicator of the total protein content of 38.39±0.000059 is shown by the method using the commercial BSA Gold Protein kit. The average values of total protein content showed the results obtained by the Bradford method and the determination of total protein by a commercial set of Clini Test-BL. The data obtained allow us to recommend the use of a set of Clini Test-BL for use in quality control, since this set is characterized by the ease and speed of the determination method, which is not inferior to the classical Lowry method. When determining the total protein content in the bulk product used to prepare the QazVac vaccine against COVID-19, the classical Lowry method showed the best result with a high degree of sensitivity.

The Influence of Silver-Containing Bionanomaterials Based on Humic Ligands on Biofilm Formation in Opportunistic Pathogens.

The uncontrolled use of antibiotics has led to a global problem of antimicrobial resistance. One of the main mechanisms of bacterial resistance is the formation of biofilms. In order to prevent the growth of antimicrobial resistance, it is crucial to develop new antibacterial agents that are capable of inhibiting the formation of biofilms. This makes this area of research highly relevant today. Promising candidates for these antibacterial agents are new bionanomaterials made from natural humic substances and silver nanoparticles. These substances have the potential to not only directly kill microorganisms but also penetrate biofilms and inhibit their formation. The goal of this study is to synthesize active pharmaceutical substances in the form of bionanomaterials, using ultradispersed silver nanoparticles in a matrix of coal humic substances, perform their characterization (NMR spectroscopy, TEM, and ICP-AES methods), and research their influence on biofilm formation in the most dangerous opportunistic pathogens (E. coli, Methicillin-resistant St. Aureus, K. pneumoniae, P. aeruginosa, St. aureus, A. baumannii, and K. Pneumonia). The results showed that all of the studied bionanomaterials had antibacterial activity against all of the opportunistic pathogens. Furthermore, they were found to have a suppressive effect on both pre-existing biofilms of these bacteria and their formation.

PREPARATION AND STANDARDIZATION OF EXPERIMENTAL BATCHES OF "PHOTORAN E6, LYOPHILIZATE FOR THE PREPARATION OF SOLUTION FOR INFUSIONS", A DRUG FOR PHOTODYNAMIC THERAPY USING HOGWEED LEAVES AS RAW MATERIALS

Introduction. In recent years, there has been a sharp increase in the number of oncological diseases associated with more frequent detection of malignant neoplasms, including in the early stages. Photosensitizers are widely used for the diagnosis and treatment of oncological diseases. Already at the beginning of the twentieth century, it was discovered that a cancer cell is able to accumulate and retain colored substances [1]. In Russia, the photosensitizer "Photoran E6, lyophilizate for the preparation of an infusion solution" is widely used for the diagnosis of malignant tumors. The sub-stance for its production is sodium chloride E6, obtained by chemical synthesis from methylpheophorbide A. which, according to the patent, is extract-ed from the biomass of the green microalgae Spirulina Platensis, native to Japan and California. In this regard, there is an urgent question of replacing imported intermediates with more affordable ones. For this purpose, Sosnowski's hogweed (Heraculum sosnowskyi) was chosen. In connection with the above, the aim of this study was to obtain and standardize in terms of quality the finished drug for photodynamic therapy "Pho-toran E6, lyophilizate for the preparation of an infusion solution" produced from sodium chloride E6, synthesized using a new type of raw material (leaves of borscht Sosnowski). Material and methods. The object of the study was three series of the finished medicine product (hereinafter referred to as GLS) "Photoran, lyophi-lizate for the preparation of an infusion solution", produced from an active pharmaceutical substance synthesized from methylpheophorbide a, ob-tained, in turn, from the leaves of borscht Sosnovsky as an alternative source of vegetable raw materials [2]. The quality indicators of the finished me-dicinal product were determined according to the methods described in the regulatory documentation in accordance with the Regulatory Documentation of the XIV and XV editions. Results. For the research, the aseptic production of three series of GLS was carried out from the previously produced active pharmaceutical sub-stance Sodium Chloride E6. AFS was synthesized from an intermediate obtained from the leaves of borscht Sosnowski. The standardization of the ob-tained series of the drug has been carried out. The possibility of using Sosnowski's hogweed as a plant raw material for the production of photosensi-tizers based on chlorophyll derivatives has been evaluated. Conclusion. In the course of the study, the technology of aseptic production of GLS "Photoran, lyophilizate for the preparation of solution for infusions" was implemented. The quality indicators of the three produced batches of the finished drug have been determined.

Modern methods for controlling and regulating impurities in pharmaceuticals

The article is dedicated to modern principles of regulation and control of impurities in pharmaceutical products and active pharmaceutical substances, which is a crucial task for the pharmaceutical industry. The relevance of this research stems from the fact that the presence of impurities in pharmaceutical products can significantly affect their safety and efficacy, making the development of stringent control methods throughout the entire life cycle of a pharmaceutical product essential. Even small amounts of impurities can cause adverse effects or reduce the therapeutic value of drugs, as evidenced by historical examples. The aim of the study is to analyze and systematize contemporary approaches to the detection and regulation of impurities in pharmaceutical products. The paper reviews current regulatory requirements, including guidelines from the The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) and regulatory documents of the Eurasian Economic Community, as well as national pharmacopeias of the Russian Federation, the United States of America, and the European Union. Conclusion. Additionally, the article emphasizes the need for harmonization of standards and practices for impurity control, which is especially relevant in the context of the globalization of the pharmaceutical market. Harmonization not only enhances the safety and efficacy of pharmaceutical products but also improves patients' quality of life by reducing the risk of adverse effects associated with impurities. The article also explores prospects for further improvement of regulatory acts and the implementation of advanced technologies aimed at enhancing standardization and quality control in the pharmaceutical industry.

Current and Future Pharmacopoeial Requirements for the Quality of Extemporaneous Medicinal Products: A Review of Regulatory Standards

INTRODUCTION. The organisation of extemporaneous compounding in pharmacies is currently considered to be a priority for the national healthcare system. Extemporaneous medicinal products used by patients should meet high quality, safety, and efficacy standards. Provision of consistent and science-based requirements for the quality of medicines, particularly those compounded in pharmacies, is a key area of concern for standardisation.AIM. This study aimed to analyse international experience and determine areas for the improvement of standardisation approaches of the State Pharmacopoeia of the Russian Federation to the quality of medicinal products compounded in pharmacies.DISCUSSION. The study investigated the common and distinctive properties of extemporaneous preparations compounded in pharmacies (and not subject to registration) and medicinal products manufactured by pharmaceutical companies. The authors analysed the requirements and recommendations for compounding and quality assurance of extemporaneous medicinal products provided by the regulatory and pharmacopoeial authorities of the Russian Federation, the Republic of Belarus, the European Union, the United Kingdom, the USA, and Canada. The United States Pharmacopeia includes circa 150 individual monographs for non-sterile compounded drug products with particular compositions and over 20 individual monographs for sterile compounded drug products. The European Pharmacopoeia requires that extemporaneous preparation should be organised within the quality assurance framework of a pharmacy after a proper risk assessment. In the Republic of Belarus, extemporaneous medicinal products are compounded in line with the requirements of the Good Compounding Practices and the State Pharmacopoeia of the Republic of Belarus, which provide rapid testing procedures for extemporaneous medicinal products.CONCLUSIONS. The study identified the following areas for the development of requirements for the quality of medicinal products compounded in pharmacies: the determination of rapid testing approaches; the development of pharmacopoeial monographs for active pharmaceutical substances used in extemporaneous compounding, with the Identification section supplemented with an additional subsection on pharmacy-specific analytical procedures; and the development of pharmacopoeial monographs for frequently used compounding formulae.

Identification and synthesis of metabolites of the new 4.5-dihydroisoxazol-5-carboxamide derivate

Introduction: 3-(2-butyl-5-chloro-1H-imidazole-4-yl)-N-[4-methoxy-3-(trifluoromethyl)phenyl]-4,5-dihydro-1,2-oxazole-5-carboxamide is new antirheumatic drug. It is necessary to identify and synthesize the biotransformation products for its complete pharmacokinetic study. Materials and Methods: A biotransformation study was carried out by intraperitoneal administration of the drug to Wistar rats and Soviet Chinchilla breed rabbits. Animal blood sampling was performed before the injection and 0.5 h, 1 h, 2 h, 4 h, 24 h after the injection of the investigated compound. The samples were immediately centrifuged for plasma separation. Urine was simultaneously collected from rats before the administration and at intervals of 0-2 h, 2-4 h, 4-6 h, 6-24 h after administration, faeces – before administration and at intervals of 0-12 h and 12-24 h after administration. The samples were analyzed by HPLC-MS/MS after immediate preparation by adding acetonitrile. Results and Discussion: 3-(2-butyl-5-chloro-1H-imidazole-4-yl)-4,5-dihydro-1,2-oxazole-5-carboxylic acid and 4-methoxy-3-(trifluoromethyl)aniline – hydrolysis products of the active substance were found during the analysis of plasma, urine and fecal samples. The 4,5-dihydro-1,2-oxazole-5-carboxylic acid derivative has been synthesized. The second metabolite is a raw material for production of active pharmaceutical substance. During comparative tests, no significant difference between the retention times, ratio areas of chromatographic peaks at the main MRM-transitions and mass spectra of these metabolites on chromatograms of standard and animal samples was found, which indicates the correct identification of biotransformation products. Conclusion: The studied drug undergoes biotransformation by hydrolysis to form two main metabolites: 3-(2-butyl-5-chloro-1H-imidazole-4-yl)-4,5-dihydro-1,2-oxazole-5-carboxylic acid and 4-methoxy-3-(trifluoromethyl)aniline. The structure of the metabolites was confirmed by comparison with the synthesized standard samples using HPLC-MS/MS.

Effect of pH on Toxicity of Zinc Sulphate Hydrate Solutions Using the Spirotox Test

Introduction: Spirostomum ambiguum is commonly used as a biological indicator due to its high sensitivity to heavy metals. It functions normally across a wide pH range of 4.5 to 8.0. Testing on protozoan organisms provides a general understanding of the impact of pH on the toxicity of zinc in pharmaceuticals. By investigating the toxicity of zinc in environment with varying pH levels, the optimal environment for zinc absorption can be determined. Objective: The objective of this study is to assess the toxicity of zinc sulphate solutions using different forms of hydrating and at various pH levels, applying the Spirotox method. Methods: An experiment was conducted using a solution of pharmaceutical substances, specifically zinc sulphate heptahydrate and zinc sulphate monohydrate, with a concentration of 0.01 M. The study aimed to investigate the effect of the solution on the lifespan of S.ambiguum within the pH range of 4.8-6.5. Results: Arrhenius activation energy (Ea linear correlates with LD50 oral toxicity for rats) decreases as pH increases from 4.8 to 6.5 for ⋅7 and ⋅ solutions. The values decrease from 140±10 kJ/mol to 55±10 kJ/mol and from 110±2 kJ/mol to 60±10 kJ/mol, respectively. Conclusion: It was found that the toxicity of zinc sulphate hydrate solution towards S. ambiguum increased as the pH value increased. Hydrating of the active pharmaceutical substance was a crucial factor, it is critically important for preparation of solutions of an active pharmaceutical substance. Keywords: zinc sulphate, Spirotox test, pH, activation energy, toxicity.

Thermal decomposition of active pharmaceutical substances and accuracy of the related kinetic predictions: The case of nifedipine

Non-isothermal thermogravimetry was used to study the thermal decomposition of the nifedipine drug. The dominant process of thermal molecular degradation (as confirmed by Raman spectroscopy) starts very slowly at ∼ 150 °C, which is below the melting point of Tm ≈ 170 °C. The decomposition kinetics was described in terms of the autocatalytic Šesták-Berggren kinetics; a novel approach denoted as single-curve multivariate kinetic analysis (sc-MKA) was used to determine the following set of parameters: activation energy of decomposition Ed = 115.5 ± 2.4 kJ mol-1, decomposition pre-exponential factor log(Ad/s-1) ≈ 8.5–8.9, Šesták-Berggren kinetic exponents M ≈ 0–0.2 and N ≈ 0.3–0.5. The consequent kinetic predictions based on these results have confirmed the good thermal stability of nifedipine, which allows for the melt-quenching preparation of the amorphous phase as well as for the processing via hot melt extrusion and 3D printing. An in-depth analysis of the relevant performance of the sc-MKA approach was done based on the akin literature data for indomethacin, nimesulide, and griseofulvin. The comparison has revealed more than sufficient performance of the sc-MKA method with regard to its application to the thermal decomposition data of active pharmaceutical substances. The most critical aspect of the sc-MKA procedure was demonstrated to be the accurate determination of the model-free parameters (activation energy Ed and pre-exponential factor Ad) and their temperature trends. Considering the specificity of the sc-MKA method, i.e., the fixed value of Ed(T), the determination of the temperature dependence of Ad(T) is of utmost importance and worth extensive repeatability checks.

Воспроизведенная комбинация модуляторов белка CFTR — ивакафтора и лумакафтора. Этические и фармакокинетические аспекты

The lack of effective and affordable therapies for rare diseases is an important ethical issue. One example is cystic fibrosis (CF), a chronic, progressive disease characterized by an impaired function of all exocrine glands. The combination of ivacaftor and lumacaftor (CFTR potentiator and corrector) should lead to a sufficient level of protein on the cell surface and to an increase in its activity, thereby correcting the impaired function. Development of a generic drug containing ivacaftor and lumacaftor as active pharmaceutical substances will increase the availability of this medication and improve patient survival. To study comparative pharmacokinetics and bioequivalence of drugs containing ivacaftor and lumacaftor in healthy volunteers. It was conducted as an open-label, randomized, crossover bioequivalence study involving a single intake of the drug during each period under fed condition in healthy male and female volunteers. The conclusion about bioequivalence was made if 90% confidence interval for primary pharmacokinetic parameters (Cmax, AUC0-t) fell within the accepted bioequivalence limits of 80–125%. According to the results of the study, it was shown that the values of 90% CI of the geometric mean of the main pharmacokinetic parameters for ivacaftor and lumacaftor fall within the acceptance limits for bioequivalence. According to the applied criteria, the drugs are bioequivalent, which makes it possible to recommend the investigational drug to the Ministry of Health of the Russian Federation for obtaining the registration status.

<i>In vitro</i> influence on fibrinolysis parameters of aqueous extracts of mushrooms growing on woody plants

Introduction. Various compounds of mushroom extracts are of interest as a source of new active pharmaceutical substances with antithrombotic activity.Aim. The aim of the work is to determine the potential antithrombotic activity in vitro of aqueous extracts of xylotrophic mushrooms: Lentinula edodes, Pholiota squarrosa, Flammulina velutipes (Curtis) Signer, Kuehneromyces mutabilis, Armillaria cepistipes, Armillaria borealis, Hypholoma capnoides, Lentinellus cochleatus, Léccinum aurantíacum, Pleurotus ostreatus.Materials and methods. The study of fibrinolytic activity of water extracts of mushrooms in vitro was carried out. The effect of test objects on plasminogen activation, on the activity of enzyms of the fibrinolysis system α2-antiplasmin, on the time of euglobulin clot lysis and on the concentration of fibrinogen was evaluated. For the analysis we used reagent kits of SPD «RENAM» and LLC Company «Тechnology-Standard» (Russia).Results and discussion. Pronounced antithrombotic activity is demonstrated by an aqueous extract of L. edodes and fractions of aqueous extract. Extracts of L. edodes affect all studied parameters of fibrinolysis to varying degrees. Caps and stems of P. squarrosa, K. mutabilis, and F. velutipes aqueous extracts and fractions of aqueous extracts also demonstrate antithrombotic (fibrinolytic) activity in vitro, having a statistically significant effect on various parameters of fibrinolysis. A less pronounced effect on fibrinolysis parameters in comparison with the above samples is observed when using an aqueous extract of L. cochleatus marked. A. cepistipes, A. borealis, H. capnoides, L. aurantiacum, P. ostreatus fruiting bodies aqueous extracts do not affect the studied parameters of fibrinolysis and the concentration of fibrinogen.Conclusion. The data obtained make it possible to recommend extracts from L. edodes, F. velutipes, P. squarrosa and K. mutabilis for further study, for example, in order to develop functional food products for cardiovascular diseases or to create antithrombotic drugs.

Modern extemporaneous pharmaceutical manufacturing: problems and prospects

Relevance. Extemporaneous manufacturing of preparations is an important component of modern patient-oriented medicine. This article provides an analysis of regulatory documents regulating the manufacture of drugs in a pharmacy, identifies problems and weaknesses in rationing production, and proposes constructive ways to solve the problems presented.Methodology. When conducting the study, methods of content analysis, comparative and logical analysis, and observation were used.Results. This article provides a critical analysis of the legal framework regulating the extemporaneous production of drugs. Weaknesses in the regulation of pharmaceutical production of dosage forms have been identified. In particular, the following problems are presented: checking the doses of active pharmaceutical substances and assessing the compatibility of prescription ingredients; issues related to the expiration dates of extemporaneously manufactured medicinal products; and problems with regulating conditions for the manufacture of drugs in pharmacies. Difficulties in equipping pharmacies with small-scale mechanization and aspects of the provision of modern active pharmaceutical ingredients and excipients are reflected, and the potential of physicians in prescribing extemporaneous prescriptions is noted.Conclusion. The proposed solutions to the identified problems will allow extemporaneous production to a new, more advanced level and expand the capabilities of personalized medicine.

Sensitive and validated UPLC-MS/MS method for the identification and characterization of forced degradation products for felodipine

AbstractA rapid, stability indicating reverse phase liquid chromatographic method was developed for the determination of purity of Felodipine in active pharmaceutical substance form in the presence of its impurity and its degradation products. To develop the method which is also compatible to liquid chromatographic mass spectroscopic technique. The developed method is also used to determine the assay of Felodipine in bulk drug form. The drug is subjected to various stress conditions like acidic, basic, oxidation, UV light and thermal conditions. Considerable degradation was observed during base hydrolysis. Two degradation products were identified. The Waters Acquity UPLC BEH C18, 2.1 × 100 mm, 1.7 µm Column was used to achieve chromatographic separation. The gradient conditions, diluent and injection volume were optimized to achieve the acceptable resolution between impurities and its degradation products from Felodipine and to get good peak shapes. The masses were determined for main compound and its identified degradation products. Further, the characterization studies for main compound and its degradation products were performed using LCMSMS Q-TOF.

ИЗУЧЕНИЕ ВАЛИДАЦИОННЫХ ХАРАКТЕРИСТИК МЕТОДИК ИСПЫТАНИЯ НА ПОДЛИННОСТЬ И КОЛИЧЕСТВЕННОГО ОПРЕДЕЛЕНИЯ АЦИЗОЛА В СТОМАТОЛОГИЧЕСКОМ ГЕЛЕ

In the quality assurance system for medicinal products, an important role is played by the procedure for validation of analytical methods at the stages of development, testing, technology and serial production. The need for validation at the stage of drug development is due to the fact that it is possible to promptly identify the shortcomings of the method and significantly improve it. The purpose of this study is to validate methods for testing «The authenticity» and «Quantitative determination» of acyzole in dental gel. Materials and methods. To achieve this goal, an active pharmaceutical substance of pharmacopoeial quality was used. When developing methods for testing for “Authenticity,” a reaction with oxalic acid to the zinc cation determined in the composition of acyzole was used. For the quantitative determination of acyzole, the complexometric method was used. The objects of the study were three series of gel samples. Results and discussion. As a result of the research, a method for testing «Authenticity» was developed; it was found that the proposed method is characterized by a negative analytical signal on model mixtures free of the analyte component and placebo, and a positive analytical signal on model mixtures of various compositions containing the analyte component. The validation characteristics of methods for the quantitative determination of acyzole in the gel were studied. positive results were obtained. The proposed methods can be used for inclusion in the regulatory documentation for the developed gel. Conclusion. When assessing the validation characteristics of the proposed methods, it was established that the methods for testing the «Authenticity» and «Quantitative determination» of acyzole in the gel are specific, the methods of quantitative determination are characterized by accuracy and repeatability, a linear dependence in the analytical range of ±20% of the declared amount of acyzole, which allows them to be used for a reliable assessment quality of the developed dental gel.

Development and validation of a method for the quantitative determination of metoprolol succinate in the air of the working area by high-performance liquid chromatography using mass spectra

Introduction. Metoprolol is a cardioselective blocker of β-adrenergic receptors. Drugs containing metoprolol have hypotensive, antianginal, antiarrhythmic effects. Moderately dangerous when taken orally, has a general toxic effect. Metoprolol is produced in two forms: in the form of salts of tartaric and succinic acids, the approximate safe level of exposure to which is 0.3 mg/m3. To ensure the safety of workers, it is necessary to carry out sanitary and hygienic air control. Methods for the quantitative analysis of metoprolol in the air of the working area, presented in the current scientific literature, allow monitoring only for metoprolol salts in the form of tartrate. 
 The aim of the study is to develop and validate an analytical method for the quantitative determination 
 of metoprolol succinate in the air of the working area by HPLC-MS. 
 Materials and methods. For the analysis, standard samples of metoprolol succinate and air samples taken at all stages of the life cycle of the production of the drug metoproolol were used. Chromatographic separation and detection were performed on an Agilent 1260 Infinity G6125B high performance liquid chromatograph (Agilent Technologies, USA) equipped with a column and sample thermostat, a degasser, a pump, an autosampler, and a G6125B mass selective detector. A Waters Xbridge® C18 3.5 µm column was used; 2.1×30 mm (Waters, Ireland). Primary data were processed using Open Lab CDS ChemStation Edition (Rev. C.01.07 SR3) software (Agilent Technologies, USA).
 Results. A method for the quantitative determination of metoprolol succinate in air samples by HPLC-MS has been developed and validated. During the validation of the developed methodology, it was found that the validation characteristics (specificity, linearity, accuracy and precision, correctness, reproducibility) are within the acceptance criteria. The analytical range of the technique was 0.1–10.0 µg/ml and allows the developed technique to be used to control the average shift MPCs in the air of the working area.
 Limitations. The results of the study of the air in the working area are applicable only to the production of the drug metoprolol in the form of tablets; the study of air during the synthesis and packaging of the active pharmaceutical substance was not carried out. 
 Conclusion. An analysis of air samples from the production of the drug metoprolol was carried out and the safety of working conditions was proved.

Optimization of the thin layer chromatography method conditions for the detection of related dioxidine impurities

When developing methods for analysing active pharmaceutical substances (API), it is necessary to take into account modern requirements for the content of impurities in them. At the same time, for some drugs that have been on the pharmaceutical market for a long time, analysis methods are still used that are unable to provide the necessary sensitivity and specificity. One of these API is 2,3-bis(hydroxymethyl)quinoxaline-1,4-dioxide (dioxidin), which has high bactericidal activity against a wide range of microorganisms. According to the requirements of the State Pharmacopoeia of the Russian Federation XIV edition, related impurities of this API are analysed by thin layer chromatography (TLC). However, this method is semi-quantitative and relative. The purpose of this work is to determination of impurities in the dioxydine substance using high-performance liquid and thin-layer chromatography methods. In accordance with the requirements of the monograph of the State Pharmacopoeia of the Russian Federation, the total content of all impurities in the API is assessed and only the content of quinoxidine is determined separately, which is often absent from the substance. For the first time, three typical dioxidine related impurities were revealed using HPLC. Next, we selected the optimal conditions for their detection by TLC. Methanol/acetonitrile (75/25) and dichloromethane/acetonitrile (50/50) mixtures were the most suitable as a solvent for impurities and the TLC elution system, respectively. Under the selected conditions, satisfactory separation of the chromatographic zones of all four impurities and the dioxidine substance was observed. The corresponding zones were clearly visible at a substance concentration of 1%. However, reducing their concentration to the required 0.1% made it difficult to visually detect impurities. Thus, even when optimizing TLC conditions, it is impossible to obtain reliable data on the content of related impurities, which is due to the limitations of the method. On the contrary, the analysis of dioxidine substances by HPLC allows fairly accurate quantitative determination of individual related impurities.

ПРИМЕНЕНИЕ СОРБЕНТА ДЛЯ ТВЕРДОФАЗНОЙ ЭКСТРАКЦИИ НА ОСНОВЕ ГИПРОМЕЛЛОЗЫ ДЛЯ КОЛИЧЕСТВЕННОГО ОПРЕДЕЛЕНИЯ КИСЛОТЫ САЛИЦИЛОВОЙ В КОМБИНИРОВАННЫХ ЛЕКАРСТВЕННЫХ ПРЕПАРАТАХ

Introduction. In quality control of pharmaceutical formulations, there are methodological approaches based on the preliminary separation of sample components and approaches without preliminary separation. The use of solid-phase extraction (SPE) allows to separate analytes during sample preparation, with the separation selectivity determining the analysis selectivity as a whole. Currently, sorbents that provide selectivity in interaction with the analyte are of interest. Various derivatives of cellulose are used to obtain selective sorbents: ethylcellulose, hydroxypropylmethylcellulose (hypromellose), carboxymethylcellulose, etc. On the pharmaceutical market of the Russian Federation, there are pharmaceutical formulations with the active pharmaceutical substance salicylic acid. The use of selective SPE for analysis is a promising direction of research in drug quality control. Aim. Evaluation of the metrological characteristics of benzocaine quantitation by UV-spectrophotometry in certain pharmaceutical formulations using hypromellose sorbent for solid-phase extraction. Material and methods. The sorbent using a method developed by us was obtained. Salicylic acid (FS.2.1.0634, Russia State Pharmacopoeia XV ed.) to form active binding sites in sorbent structure was applied. The sorbent structure is polycyanoacrylate matrix with hypromellose fragments, the surface area is 255.50 m2/g, pore volume is 0.1433 cm3, pore diameter is 5.32 nm. The sorption capacity of hypromellose sorbent for benzocaine was 12.2 ± 0.8 μg/g. The SPE technique is proposed. SPE includes the stages: conditioning, sample addition and step-by-step elution with purified water and sodium hydroxide solution 0.1 mol/L. The spectrophotometer SF-56 was used to measure the absorbance at an analytical wavelength of 286 nm. To evaluate the selectivity (specificity) of sample preparation during chromatographic (SPE) separation of pharmaceutical formulations, absorption spectra of eluates in the wavelength range of 200-400 nm were scanned. The salicylic acid identification in the eluate was carried out based on absorption peaks at 228 and 298 nm.The salicylic acid calibration curve in the range of 1-20 μg/ml. To determination the metrological characteristics of salicylic acid quantitation method, 16 parallel determinations of samples of each pharmaceutical formulation were made. Statistical processing of the experimental results and determination of the metrological characteristics evaluation of analytical technique were carried out in accordance with general pharmacopoeial monograph (OFS.1.1.0013 "Statistical processing of results of physical, physicochemical, and chemical tests"). Results. When realizing the salicylic acid quantitation technique for measuring absorbance, it is advisable to use only the first portions of eluate (a solution of sodium hydroxide) in a volume of 5-10 ml were obtained. The relative error in determining the average content of salicylic acid in the analyzed preparations ranged from 1.18 to 1.44% for methods with solid-phase extraction (SPE) and 1.86-2.62% in its absence. It has been shown that there is a statistically significant difference in reproducibility for all preparations, as indicated by F-criteria values ranging from 7.81 to 13.5, exceeding their tabulated values. For the method without solid-phase extraction (SPE), there was an overestimation of the salicylic acid content in all cases, increasing the systematic error on average by 87.5%. Conclusion. The possibility of using a selective hypromellose sorbent for sample preparation for salicylic acid quantitation determination in pharmaceutical formulations has been showed. The use of sorbent for analysis provides to systematic error decrement of salicylic acid quantitation by an average of 87.5%.

Methodological and regulatory aspects of pharmaceutical development of biological products

The aim of the work was to conduct an analysis of the current state and current trends in the approval of drugs, as well as some aspects of the methodology for their development based on biological molecules and registration.Materials and methods. The material for the analysis was taken from the abstract databases of PubMed, Google Scholar and e-library.ru. The search was carried out using publications for the period from 2008 to 2023, the keywords were as follows: “biologicals”, “new drug approval”, “drug authorization”, “drug development”, “biosimilar”, taking into account various spellings.Results. Over the past 15 years, scientists have been observing revolutionary trends and processes in the field of the drug development, especially biopharmaceuticals. Significant advances have been made in gene, immune and cell therapies, resulting in the approval of such drugs more than doubling over the past ten years. The development of biological drugs includes the identification and testing of molecular targets and requires a deep understanding of the structure and functioning of the polypeptides involved in the development of the effect. The features of these active pharmaceutical substances are a high molecular weight, a complex three-dimensional structure and a high immunogenic potential. Preclinical and clinical studies of biologics have unique challenges. Selecting appropriate animal species, understanding the immunogenicity, and assessing pharmacodynamics and toxicological properties require a multilevel, detailed approach. The article discusses the regulatory framework under which these drugs are registered, summarizing the guidelines provided by international organizations such as the International Council for Harmonization and various national agencies.Conclusion. The analysis highlights the current advances and prospects in the development of biologics, highlighting their key role in future transformations in the treatment of rare diseases and oncology, approaching the era of personalized medicine. Monitoring the development directions and technological approaches, as well as the commitment to global methodological and regulatory aspects can become a catalyst in the development of the Russian pharmacology.

On the cardiovascular effects mechanism of humic nature macromolecular compounds

Introduction. The search for new drugs to reduce effectively myocardial damage during ischemia and reoxygenation is relevant considering that many pharmacological drugs have a number of limitations for patients and their use is accompanied by negative side effects.Aim: To assess the NO-synthase possible role in the cardiovascular effects mechanism of the standardized active pharmaceutical substance (AFS) of the humic acids (HA) isolated from peat.Material and Methods. The experiments were carried out on the isolated perfused rat heart model using the Langendorff method. We studied the effect of a standardized sample of peat HA at concentrations of 0.001; 0.01; 0.1 mg/ml on coronary perfusion rate and myocardial contractility parameters. To assess the NO-synthase value in the HA effects realization, the enzyme was pre-inhibited using L-NAME (10 µM/L) before the test sample addition.Results. HA sample using contributed to an effective increase in the heart coronary perfusion rate due to the NO-synthase signaling mechanism activation. There was some decrease in contractility and end-diastolic pressure associated with the NOsynthase activation because the enzyme inhibition with L-NAME removed all effects of the test HA sample.Conclusion. The obtained data show to the peat HA have vasodilating properties associated with the NO-synthase activation. The such effect presence indicates on the prospects for further investigation of these compounds cardiotropic properties in the order developing new effective means for improving intracardiac hemodynamics and limiting Ca2+ overload of cardiomyocytes in conditions of ischemia and reperfusion.

Determination of the Active Pharmaceutical Ingredients in Saridon Tablets Using an Economical and Sensitive Thin Layer Chromatography Method Combined with Densitometry

A new TLC–densitometric method has been developed for the identification and quantification of paracetamol (PA), propyphenazone (PP) and caffeine (C) in Saridon tablets using the NP-TLC technique combined with densitometry. This method allows for the simultaneous determination of PA, PP, and C in the same sample. Among all the tested chromatographic conditions, the mixture consisting of chloroform + toluene + ethyl acetate + ethanol + acetic acid (18:18:7.5:5.0:0.3, v/v/v/v/v) and a silica gel 60F254 plate proved to be the most effective for the separation of the three tested active pharmaceutical ingredients (APIs) and substances related to paracetamol. The full validation of the proposed NP-TLC method proved that it is specific, precise, accurate, robust and sensitive. The percentage content in relation to the content declared by the manufacturer was for propyphenazone 99.8%, paracetamol 101.6% and caffeine 100.8%, which was in accordance with pharmacopoeial requirements. The results presented indicate the possibility of using the developed method in the routine control of pharmaceutical preparations containing these APIs. The proposed method is economical and more sensitive compared to the previously proposed planar methods for the simultaneous determination of APIs. What is more, the presented method may be an excellent economical alternative when the HPLC method is unavailable for such a determination.