Single-Dose Bioequivalence Study of Rivaroxaban-Containing Medicinal Products in Healthy Volunteers

Therapeutically, new oral anticoagulants (NOACs) are considered to be non-inferior or superior to vitamin K antagonists (warfarin). NOACs are included in current guidelines for the treatment of various cardiovascular diseases. Rivaroxaban medicinal products have been shown to effectively fight thrombotic complications of the new coronavirus infection, COVID-19. The wide clinical use of rivaroxaban products motivates the development of generics.The aim of the study was to compare the pharmacokinetics and safety of rivaroxaban medicinal products in a single-dose bioequivalence study in healthy volunteers under fasting conditions.Materials and methods: the bioequivalence study compared single-dose oral administration of Rivaroxaban, 10 mg film-coated tablets (NovaMedica Innotech LLC, Russia), and the reference product Xarelto®, 10 mg filmcoated tablets (Bayer AG, Germany), in healthy volunteers under fasting conditions. The open, randomised, crossover trial included 46 healthy volunteers. Each of the medicinal products (the test product and the reference product) was administered once; blood samples were collected during the 48 h after the administration. The washout between the study periods lasted 7 days. Rivaroxaban was quantified in plasma samples of the volunteers by high performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS).Results: no adverse events or serious adverse events were reported for the test and reference products during the study. The following pharmacokinetic parameters were obtained for Rivaroxaban and Xarelto®, respectively: Cmax of 134.6 ± 58.0 ng/mL and 139.9 ± 49.3 ng/mL, AUC0–48 of 949.7 ± 354.5 ng×h/mL and 967.6 ± 319.9 ng×h/mL, AUC 0–∞ of 986.9 ± 379.7 ng×h/mL and 1003.6 ± 320.4 ng×h/mL, T1/2 of 8.2 ± 3.2 h and 7.8 ± 3.3 h. The 90% confidence intervals for the ratios of Cmax, AUC0–48, and AUC0–∞ geometric means were 88.04–108.67%, 89.42–104.92% and 89.44–104.81%, respectively.Conclusions: the test product Rivaroxaban and the reference product Xarelto® were found to have similar rivaroxaban pharmacokinetics and safety profiles. The study demonstrated bioequivalence of the medicinal products

Актуальные направления применения клеточной терапии в регенеративной медицине

Cell therapy is a key tool of regenerative medicine, but until the beginning of the last decade, products based on viable human cells were used primarily to repair damaged tissues and organs. Currently, the field of application of biomedical cell products has expanded significantly, but researchers still show considerable interest in the use of human cells in regenerative medicine. The stage of development of cell products varies significantly depending on the type of tissue and pathology, and ranges from preclinical and pilot clinical trials to authorised drugs with a long history of use. On the one hand, this may be attributed to methodological differences in the production and use of cell products, and on the other, to specific aspects of differentiation of cell types used in regenerative medicine, primarily mesenchymal stem cells. The aim of this study was to analyse current trends in the use of cell therapy in regenerative medicine and prospects for using available technologies. The paper summarises the main achievements in the use of cell therapy for regeneration of skin, bone and cartilage, nervous and cardiovascular systems. The key mechanisms of cell therapy effect are determined, on the one hand, by the differentiation potential of multipotent cells, and on the other, by the complex (immunomodulating, angiogenic, proliferative) action of the proteome expressed by the administered cells. The paper describes viable cell-based products currently authorised for each indication, and analyses the level of their clinical use. It might be promising to use directed cell differentiation technologies, as well as induced pluripotent cells in regenerative medicine.Клеточная терапия является ключевым инструментом регенеративной медицины, и вплоть до 2010 года продукты на основе жизнеспособных клеток человека применялись преимущественно для восстановления поврежденных тканей и органов. В настоящее время сфера применения биомедицинских клеточных продуктов значительно расширилась, однако интерес исследователей к использованию клеток в регенеративной медицине остается стабильно высоким. В зависимости от типа ткани и патологии уровень разработки клеточных продуктов значительно отличается: от доклинических и пилотных клинических исследований до зарегистрированных препаратов с многолетним опытом применения. Данный факт, с одной стороны, может быть связан с методологическими особенностями производства и применения клеточных продуктов, с другой — с особенностями дифференцировки типов клеток, используемых в регенеративной медицине, прежде всего мезенхимальных стволовых клеток. Цель работы — анализ современных направлений использования клеточной терапии в регенеративной медицине и перспектив применения существующих технологий. Представлены основные достижения использования клеточной терапии в регенерации кожи, костно-хрящевого аппарата, нервной и сердечно-сосудистой систем. Ключевые механизмы лечебного действия клеточной терапии обусловлены, с одной стороны, потенциалом мультипотентных клеток к дифференцировке, с другой — комплексным (иммуномодулирующим, ангиогенным, пролиферативным) действием экспрессируемого протеома вводимых клеток. Для каждого из показаний описаны зарегистрированные в настоящее время продукты на основе жизнеспособных клеток человека и проанализирован их уровень применения в клинической практике. Перспективным представляется использование в регенеративной медицине разработанных технологий направленной дифференцировки, а также применение индуцированных плюрипотентных клеток.

Применение проточной цитометрии для оценки качества биомедицинских клеточных продуктов

Flow cytometry is the most common method of identification and quantitation of cell surface markers. Flow cytometry can be used for  cell counting and characterization of cell types and subtypes by  labeling cells with fluorochrome-conjugated monoclonal antibodies.  Manufacturers of human cell-based medicinal products have accumulated significant experience in flow cytometry and  developed a large number of procedures that can be validated and  included into cell products specifications. The present review  summarises the experience gained with the use of flow cytometry for characterization of human cell lines used to develop cell therapy  products. Since all biomedical cell products (BMCPs) have a cellular  component, it will be necessary to use the flow cytometry method for identification testing of BMCPs during evaluation of their quality.Метод проточной цитометрии — наиболее информативный метод идентификации и количественного определения поверхностных маркеров клеток. Проточная цитометрия дает  возможность проводить подсчет клеток, а также характеризацию их типов и подтипов путем  мечения клеток моноклональными антителами, конъюгированными с флуорохромом. В  настоящее время производителями продуктов на основе клеток человека накоплен  значительный опыт применения проточной цитометрии, разработано большое количество  методик, подлежащих валидации и включению в спецификацию на клеточный продукт. В  обзоре авторами рассмотрен опыт применения метода проточной цитометрии для оценки  качества клеточных линий человека, используемых, в частности, для создания препаратов с целью применения в клеточной терапии. Учитывая обязательное наличие клеточного  компонента в составе биомедицинских клеточных продуктов (БМКП), метод проточной  цитометрии будет являться обязательным при подтверждении подлинности в ходе экспертизы качества БМКП в Российской Федерации

Международный опыт нормативно-правового регулирования препаратов, содержащих жизнеспособные клетки человека

The intensive development of cellular technologies stipulates the introduction at the global level of medicinal products based on viable human cells, which in most countries are referred to as biomedical cell products. The authors conducted a comparative analysis of the regulatory framework in different countries and determined special aspects of regulation of cell therapy products (analogues of biomedical cell products). Some countries have mechanisms for priority review of cell therapy products for marketing authorization, such as accelerated assessment, accelerated approval, or conditional marketing authorisation. These mechanisms are currently absent in Russia, because of the novelty of the regulatory framework, and the biological properties of innovative cell products. Biomedical cell products are regarded as a separate class of medicinal products in Russia, they are not treated as biologicals and are regulated by the Federal Law No. 180-FZ «On Biomedical Cell Products» of June 23, 2016. The main difference in regulation of cell-based products in the Russian Federation is the principle of unified requirements for marketing authorisation of autologous, allogeneic, and combined biomedical cellular products, and the absence of the «hospital exemptions» mechanism that exists in many countries. This mechanism allows prescription and use of personalised autologous medicines produced in the laboratory of a medical institution for a particular patient.Интенсивное развитие клеточных технологий обусловливает внедрение в мировую медицинскую практику препаратов на основе жизнеспособных клеток человека, которые в большинстве стран определяются как биологические лекарственные препараты. Авторами проведен сравнительный анализ нормативно-правовой базы разных стран мира и определены особенности регулирования препаратов для клеточной терапии (аналогов биомедицинских клеточных продуктов). В некоторых странах существуют механизмы приоритетного рассмотрения препаратов для клеточной терапии для вывода на рынок, например процедуры ускоренного рассмотрения, ускоренного утверждения, условной регистрации. Учитывая новизну нормативной базы и биологические особенности инновационных препаратов — биомедицинских клеточных продуктов, в Российской Федерации подобные механизмы в настоящее время отсутствуют. Биомедицинские клеточные продукты в России являются отдельным классом медицинских средств, отличным от биологических лекарственных препаратов, и регулируются Федеральным законом № 180-ФЗ «О биомедицинских клеточных продуктах» от 23 июня 2016 г. Основным отличием регулирования клеточных препаратов в России является принцип единых требований вывода на рынок аутологичных, аллогенных и комбинированных биомедицинских клеточных продуктов и отсутствие механизма «исключения для больничного производства» (hospital exemptions), действующего во многих странах и заключающегося в допущении применения персонифицированного аутологичного препарата, произведенного в конкретной лаборатории при медицинской организации для определенного пациента по назначению конкретного врача

SORPTION-CHROMATICITY DETERMINATION OF ACTIVE INGREDIENTS IN DRUG SUBSTANCE TETRACYCLINE HYDROCHLORIDE AND IN CYANOCOBALAMIN SOLUTION FOR INJECTION

The simple and available technique of determination of tetracycline in the form of colored complex with iron(III) ions and B12 vitamin in the form of the colored thiocyanate complex of cobalt(II) after a chemical mineralization of cyanocobalamin is offered. The analytes were separated from the accompanying components by sorption to polyurethane foam based on ethers. The conditions of sorption separation and measurement of analytical signal of these substances are optimized. The obtained results are in satisfactory agreement with data declared by the manufacturer. The adequacy of proposed approach is confirmed by results of indirect X-ray fluorescence analysis of tetracycline drug substance and injection solution B12 vitamin (cyanocobalamin)

Digital Colorimetry of Non-steroidal Anti-inflammatory Drugs: Identification Using Principal Component Method

Introduction. Digital colorimetry is one of the available and simple methods that can be used for the rapid detection of low-quality drugs. The main limitation of the method is its lack of selectivity. To increase the selectivity, the use of molecular sensors is proposed. Molecular sensors are substances that change color during physicochemical interaction with the analyte. Digital colorimetric analysis using a set of sensors allows one to obtain a large amount of information about the sample, however, such a significant amount of data is rather difficult to interpret and use for rapid assessment of the composition of the drug. In addition, the use of a large set of sensors significantly increases the level of information noise. To reduce the influence of the noise component, as well as to reduce the dimensionality of the data, it is advisable to use chemometric algorithms, in particular, the method of principal components (principal component analysis, PCA). It is shown that the using of PCA will make it possible to replace 24 values of the luminosity of color channels with 2-3 numerical values of the main components without loss of analytical information.Aim. Aim of our investigation is the development of a new approach to identifying non-steroidal anti-inflammatory drugs using multisensory digital colorimetry by the principal component method.Materials and methods. The analysis was performed in 96-well transparent polypropylene plates with flat bottom (Thermo Fischer Scientific, USA, № 430341). 100 μl of the correspond-ing sensor and 100 μl of alcohol solutions of non-steroidal anti-inflammatory substance sub-stances were consistently added to the wells of the plate. Sensor solutions were added to a separate row of wells for comparison without adding substance solutions (intact wells). After adding solutions of the substance, the plate was sealed with a film, shaken on a PST-100HL plate shaker (BioSan, Latvia) for 5 minutes and left for 20 minutes to complete the reaction. To obtain raster images an Epson Perfection 1670 office flatbed scanner (CCD matrix) with a removable cover was used. The difference in the lightness of the color channels between the analyte well and the intact well was used as an analytical signal. The obtained digital images of the cells were processed in the ImageJ program using the RGB 24 bit color model (8 bits per channel).Results and discussion. It is shown that the use of chemometric algorithms for processing the results of multisensor colorimetric analysis allows to use the entire data array in obtaining an-alytical information, and not just the lightness values of individual channels of some sensors. The method of principal components allows you to simultaneously get rid of the noise com-ponent of the colorimetric signal and highlight the most sensitive sensors for this sample. The adequacy of the proposed combined approach is confirmed by the identification of active sub-stances in 5 drugs of the group of non-steroidal anti-inflammatory drugs.Conclusion. The approach proposed in this work can be successfully applied as an express and available way to assess the authenticity of medications of the group of non-steroidal anti-inflammatory drugs

Current Trends in the Use of Cell Therapy in Regenerative Medicine

Cell therapy is a key tool of regenerative medicine, but until the beginning of the last decade, products based on viable human cells were used primarily to repair damaged tissues and organs. Currently, the field of application of biomedical cell products has expanded significantly, but researchers still show considerable interest in the use of human cells in regenerative medicine. The stage of development of cell products varies significantly depending on the type of tissue and pathology, and ranges from preclinical and pilot clinical trials to authorised drugs with a long history of use. On the one hand, this may be attributed to methodological differences in the production and use of cell products, and on the other, to specific aspects of differentiation of cell types used in regenerative medicine, primarily mesenchymal stem cells. The aim of this study was to analyse current trends in the use of cell therapy in regenerative medicine and prospects for using available technologies. The paper summarises the main achievements in the use of cell therapy for regeneration of skin, bone and cartilage, nervous and cardiovascular systems. The key mechanisms of cell therapy effect are determined, on the one hand, by the differentiation potential of multipotent cells, and on the other, by the complex (immunomodulating, angiogenic, proliferative) action of the proteome expressed by the administered cells. The paper describes viable cell-based products currently authorised for each indication, and analyses the level of their clinical use. It might be promising to use directed cell differentiation technologies, as well as induced pluripotent cells in regenerative medicine

Design of preclinical studies of biomedical cell products: characteristics, key principles and requirements

According to the Federal Law «On Biomedical Cell Products», preclinical trials are an integral part of biomedical cell product (BCP) development. This article describes the basic principles of fulfilling requirements laid down in the Rules for conducting preclinical trials of BCPs. The main objective of preclinical trials is evaluation of efficacy, safety and biodistribution of cell products. Properly justified markers of biological activity must be used for reliable identification of BCP pharmacodynamic action in the host organism. BCP safety assessment must be comprehensive and include identification, characterization and quantitative evaluation of potential local and systemic toxicity, estimation of the onset of toxicity and possibility of its reduction, and the effect of a particular drug dose on the results of toxicity studies. The ultimate goal of preclinical toxicity studies is to obtain data sufficient for making a conclusion on the possibility of conducting clinical trials of BCP and determining associated risks. The key principles of preclinical trials design are a rational approach and justification of all decisions made during the study. The results of preclinical trials that were conducted in compliance with the law, can be included in the BCP dossier and considered during the product authorization by the expert institution of the Ministry of Health

International Practice of Storing Human Cell Lines Intended for Clinical Use

Currently, the Russian Federation does not have a well-established state-controlled market for cell banks (CB) containing cell material that is potentially applicable for clinical purposes. Cryopreservation of cells in cell bank (CB) is an important step in the production of a number of biomedical cell products and makes it possible to overcome difficulties faced by manufacturers during production and storage of large amounts of cell material. At present there are a large number of human cell lines in the world, which are stored in CB owned by commercial and public organisations in different countries. In addition, new cell lines are being banked every year. All this makes it difficult to find cell material suitable for production purposes or that could potentially be used as donor material in clinics. This study analysed the international practice of storing human cell lines for clinical use. The authors of the study systematised the existing CB worldwide and analysed regulatory documents governing the activities of these banks in different countries. The analysis revealed a trend towards formation of CB, often specialising in certain types of cells, as well as a trend towards creation of registries giving full information about cell lines including data on their scientific application. The increasing development and clinical use of cell therapy products in the Russian Federation and abroad will most likely lead to the increase in the number of CB and registry systems, as well as amounts of materials stored in them, including cell lines intended for clinical use

Use of Flow Cytometry for Quality Evaluation of Biomedical Cell Products

Flow cytometry is the most common method of identification and quantitation of cell surface markers. Flow cytometry can be used for  cell counting and characterization of cell types and subtypes by  labeling cells with fluorochrome-conjugated monoclonal antibodies.  Manufacturers of human cell-based medicinal products have accumulated significant experience in flow cytometry and  developed a large number of procedures that can be validated and  included into cell products specifications. The present review  summarises the experience gained with the use of flow cytometry for characterization of human cell lines used to develop cell therapy  products. Since all biomedical cell products (BMCPs) have a cellular  component, it will be necessary to use the flow cytometry method for identification testing of BMCPs during evaluation of their quality