Approaches to Pharmaceutical Analysis of Modern Peptide and Oligonucleotide Products as Illustrated by a Small Interfering RNA-Based Novel Therapeutic for the Treatment of Bronchial Asthma

Methods used to control the quality of peptide products depend on the level of development of analytical and bioorganic chemistry, and the level of instrumentation. Peptide identification is a difficult task and largely depends on the complexity of its structure. There does not exist a comprehensive and simple test, except for NMR, which, however, is rather expensive and time-consuming and involves complex data interpretations. Moreover, it does not allow for unambiguous determination of the peptide purity and formula (amino acid composition, sequence, chirality of amino acid residues). For this reason, a combination of methods is often used, including amino acid analysis, TLC/HPLC and mass spectrometry, and, less frequently, sequencing. Current international practice of peptide analysis is to use HPLC in combination with mass spectrometric, mainly tandem (HPLC-MS/MS), detection. According to literature sources the amino acid sequence of linear peptides can be analysed using various enzymes and subsequent identification of proteolysis products by mass spectrometry. This article presents approaches to the development of test methods for analysis of purity and identification testing of a small interfering RNA-based novel medicinal product, which will help standardise and control the quality of the production process

Подходы к фармацевтическому анализу современных пептидных и олигонуклеотидных препаратов на примере инновационного препарата на основе малой интерферирующей РНК для лечения бронхиальной астмы

Methods used to control the quality of peptide products depend on the level of development of analytical and bioorganic chemistry, and the level of instrumentation. Peptide identification is a difficult task and largely depends on the complexity of its structure. There does not exist a comprehensive and simple test, except for NMR, which, however, is rather expensive and time-consuming and involves complex data interpretations. Moreover, it does not allow for unambiguous determination of the peptide purity and formula (amino acid composition, sequence, chirality of amino acid residues). For this reason, a combination of methods is often used, including amino acid analysis, TLC/HPLC and mass spectrometry, and, less frequently, sequencing. Current international practice of peptide analysis is to use HPLC in combination with mass spectrometric, mainly tandem (HPLC-MS/MS), detection. According to literature sources the amino acid sequence of linear peptides can be analysed using various enzymes and subsequent identification of proteolysis products by mass spectrometry. This article presents approaches to the development of test methods for analysis of purity and identification testing of a small interfering RNA-based novel medicinal product, which will help standardise and control the quality of the production process.Используемые методы для контроля качества лекарственных средств пептидной природы зависят от уровня развития аналитической и биоорганической химии и развития приборной базы. Анализ подлинности пептида представляет собой непростую задачу и во многом зависит от сложности его структуры. Не существует однозначного и достаточно простого теста, за исключением ЯМР, который, однако, является дорогостоящим и длительным методом со сложной интерпретацией данных. Причем этот метод не позволяет однозначно установить чистоту и формулу пептида (аминокислотный состав, последовательность, хиральность аминокислотных остатков). По этой причине нередко используется комбинация методов, включая аминокислотный анализ, ТСХ/ВЭЖХ и масс-спектрометрию и, более редко, секвенирование. В мировой практике для исследования пептидов наиболее распространен метод ВЭЖХ в сочетании с масс-спектрометрическим, преимущественно тандемным (ВЭЖХ-МС/МС) детектированием. Для установления аминокислотной последовательности линейных пептидов описано применение различных ферментов с последующей идентификацией продуктов протеолиза масс-спектрометрически. В данной статье представлены подходы к разработке методик определения подлинности и чистоты инновационного лекарственного препарата пептидной природы на основе малой интерферирующей РНК с целью стандартизации и контроля качества на производстве

Use of combined drugs in treatment of respiratory viral infections

Diseases caused by viral infections of the respiratory tract occupy the first place in prevalence among all viral pathologies. Pathogens such as influenza virus, respiratory syncytial virus, rhinovirus and others that are dangerous not only due to their direct negative effect on the human body, but by the fact that they strengthen the course of other inflammatory diseases of the respiratory tract, for example bronchial asthma. Development of new molecular research methods in immunology allowed revealing in detail the peculiarities of the life cycle of certain respiratory viruses, and understanding of the mechanisms of antiviral defense. According to modern concepts the immune system is a delicate and complicated mechanism of the antiviral response, in which one of the main roles is played by cells containing on their surface CD4-receptors. Also many studies proved the key role of interferon I, II and III type as the first line of antiviral defense. The knowledge accumulated as of today in the field of virology and immunology allows us to implement a comprehensive approach in the development of new antiviral drugs with the aim to achieve greater efficiency. This approach was successfully implemented with the creation of the drug Ergoferon containing a combination of affinity purified antibodies to CD4, interferon-γ and histamine in a releaseactive form with antiviral, anti-inflammatory and antihistamine activity. Influence on the CD4-receptors enables the drug to activate the cascade of immunologic responses aimed at improving the protective properties of the body – to fight infectious agents, primarily viral in origin. Effects on the IFN system make an equivalent contribution to the effect on CD4-receptor to the antiviral properties of the drug. A unique combination of properties, high level of evidence of clinical efficacy, good tolerability and no adverse drug effects can refer Ergoferon to effective and safe drugs for treatment of the main respiratory viral infections

Development of blast-resistant rice varieties based on application of DNA technologies

Diseases of agricultural crops are the main reason for decreased yield and quality of product. Blast (causative agent: Pyricularia oryzae Cav.) is the most harmful disease on rice fields. Economic damage caused by the pathogen is significant in all areas of the world’s rice cultivation. The most effective, economically justified and environmentally friendly strategy for combating it is development of resistant varieties. The application of DNA markers linked to loci of resistance to blast is relevant in this area. This makes it possible to significantly shorten the breeding process and promptly develop disease-resistant rice forms. In this regard, the aim of the work was to develop source material for breeding as well as highly productive rice varieties and lines with genes of resistance to blast based on the use of molecular marking method. To achieve this goal, we have launched a program since 2007 aimed at introduction of the blast resistance Pi-ta gene, effective for the south of Russia, into the domestic rice cultivar Flagman. After a number of recurrent crosses, the breeding material was obtained, which was studied for economically valuable traits in breeding nurseries. As a result of evaluation and rigorous discarding as well as according to the results of a phytopathological test for blast resistance, several lines were identified that have high indicators of milled rice quality, resistance to blast, yield and economically valuable traits. Rice accession KP-171-14 with the Pi-ta gene, adapted to growing conditions in the south of Russia, resistant to the Krasnodar population of P. oryzae, and having high yield and grain quality, in 2017 was submitted for State Variety Trials (SVT) under the name Alyans. Accessions KP-30 and KP-23 are tested for economically valuable traits and disease resistance in competitive variety trials. The best accession will be submitted to SVT. The introduction and cultivation of such varieties will reduce the use of chemical protection products, obtain environmentally friendly agricultural products and avoid contamination of grain ecosystem