The human respiratory syncytial virus (RSV) is one of the most common viral pathogens that affects the lower respiratory tract and could be a reason of bronchiolitis and/or pneumonia. Currently, there are no available effective ways of treating the RSV infection. Attempts to develop preventive vaccine have been unsuccessful. The only therapeutic agent used for RSV treatment is virazole (ribavirin); however, it induces adverse effects. Medications based on neutralizing monoclonal antibodies, such as IGIV (Respigam), palivizumab (Synagis), and MEDI-524 (Numab), are under clinical trials; however, their use will be limited by their high cost. One of the promising approaches for antiviral therapy is the use of natural peptides (defensins and cathelicidins), or their synthetic analogs. The majority of currently described antiviral peptides are developed against the human immunodeficiency virus, the herpes simplex virus, and the influenza virus. At the same time, a body of experimental data evidencing anti-RSV activity of peptides has been accumulated. The main advantages of peptide drugs are their wide spectrum of antiviral activity and low toxicity. However, there are obstacles in implementing peptide-based drugs in clinical practice. Due to their low resistance to the action of serum proteases, most authors consider peptides promising only for local application. Given that RSV affects the epithelium of the respiratory tract, where the protease activity is lower than in the systemic circulation, it is possible to develop locally active peptide drugs, for example, as inhalation forms. Their stability could also be increased by the synthesis of dendrimer peptides and by the development of recombinant peptides as precursor proteins. Anti-RSV peptides can be divided into several groups: (1) attachment and/or fusion blockers; (2) peptides displaying direct virucidal activity, disrupting the viral envelope. Such peptides, which suppress early stages of the viral life cycle, are considered prophylactic agents. However, for several peptides, their immunoregulatory properties have been described, which opens the possibility for therapeutic use. This review summarizes the information on the antiviral properties of such peptides and mechanisms of their action and describes the prospects of the future development of antiviral peptides.

The composite Y14/siUTR, a complex consisting of cationic lipopeptide Y14 as an excipient and pharmaceutical substance of small interfering RNAtargeted against the UTR region of hepatitis C virus (siUTR), was investigated. The composite was intended to inhibit the hepatitis C virus replication cycle. The present work was aimed at developing pharmaceutical analytical methods for the components of this composite using HPLC-UV and UV spectroscopy.

Bronchial asthma is a chronic inflammatory disease of the airways that is characterized by episodes of shortness of breath, expiratory dyspnea, cough, wheezing, and pulmonary emphysema. At the present time, asthma is a global public health problem and affects about 5% of the worldwide population. Although a wide range of anti-inflammatory drugs is available, uncontrolled or poorly controlled asthma is still a problem, requiring the development of novel therapeutic approaches. Intense studies of the molecular mechanisms of asthma in transgenic animals performed since the 1990s implicated cytokines, such as IL-4, IL-5, and IL-13, and their receptors in the initiation and maintenance of asthma. These findings led to anticytokine therapy as a novel approach for bronchial asthma treatment. To date, many preclinical and clinical studies have been performed in this field especially with drugs based on humanized monoclonal antibodies, soluble receptors, peptide inhibitors, etc. The review summarizes the data from preclinical and clinical studies of anti-cytokine therapeutics in humans.

For evaluation of effects of release-active antibodies to CD4 on cultured lymphocytes from human peripheral blood, we measured intracellular content of lck-kinase cell-based ELISA. In cells treated with release-active antibodies to CD4, the content of intracellular lck-kinase significantly (p < 0.01) decreased in comparison with the control (purified water processed in a similar way). Phytohemagglutinin had no effect on the concentration of lck-kinase in cells. The decrease in the content of CD4-associated lck protein suggests that the preparation enhanced intracellular coupling of lck-kinase with T-cell receptor and potentiated T-cell immune response.

Asthma is among the most common chronic disorders of airways, which affects both children and adults. Asthma being a common disease among different segments of population, it has a high mortality rate and, in the absence of appropriate care, affects the quality of life and leads to economics losses. In a view of continuing growth in the incidence of asthma, it is important to find relevant biological targets for developing new approaches to astma therapy. Recent advances in molecular immunology, genetics, and bioinformatics allowed genes involved in the pathogenesis of asthma to be identified, which provided prerequisites for the development of new types of drugs that can regulate the activity of pathogenically significant genes. To date, a number of technologies for sequence-specific gene regulation (ASO, ribozymes, DNAzymes, EGS, DNA-decoys, U 1-adapters) are available, but RNA interference is the most promising approach in both terms of efficacy and financial cost. This review focuses on the generalization and analysis of experimental data regarding the use of RNA interference technology for the treatment of astma.

Antiviral activity of Ergoferon was studied in vitro on an experimental model of rotavirus infection in MA-104 cell line. In infected cells treated with Ergoferon, rotavirus titer was shown to decrease by 83 and 90% in comparison with cells treated with solvent used for Ergoferon preparation (p<0.05) and distilled water (p<0.05), respectively. These findings demonstrate high anti-rotavirus activity of Ergoferon.

Бронхиальная астма является одним из самых распространенных хронических воспалительных заболеваний дыхательных путей, от которого страдают как взрослые, так и дети. Данное заболевание распространено среди разных слоев населения, имеет высокий уровень смертности, а при отсутствии квалифицированной помощи приводит к ухудшению качества жизни населения и экономическим потерям для государства. Ввиду продолжающегося роста заболеваемости бронхиальной астмой остается актуальной задача поиска значимых биологических мишеней с целью разработки новых способов терапии. Недавние успехи в области молекулярной иммунологии, генетики и биоинформатики позволили выявить гены, вовлеченные в патогенез бронхиальной астмы, что создало предпосылки для появления лекарственных препаратов нового типа, способных регулировать активность патогенетически значимых генов. К настоящему моменту известен ряд технологий для сиквенс-специфической регуляции генов (АСО, Рибозимы, ДНКзимы, EGS, ДНК-ловушки, U1-адапторы), однако использование феномена РНК-интерференции является наиболее перспективным с точки зрения эффективности и экономической целесообразности. Данный обзор посвящен обобщению и анализу экспериментальных данных по использованию технологии РНК-интерференции для терапии бронхиальной астмы.

Activation of proinflammatory cytokines including IL-4, IL-5, IL-9 and IL-13 plays an important role in the induction and maintenance of the majority of the allergic diseases associated with allergic inflammation. Interleukin-4 makes an especially important contribution to pathogenesis of allergic conditions, such as bronchial asthma. Specifically, IL-4 provides a switch of antibody synthesis from IgM to IgE; moreover, it functions as the growth factor for mast cells and basophils. Successful attempts to create therapeutic preparations that suppress IL-4 activation by monoclonal antibody have been reported. However, these investigations were not continued. Therefore, the search for the methods to effectively suppress IL-4 remains a topical problem. The recent discovery of RNA-interference (RNAi) phenomena provides new possibilities for the regulation of gene expression. The activation of RNAi requires the use of siRNA molecules (short interfering RNAs) the sequence of which is 100% identical with that of the respective mRNA-target. In the present study, we designed siRNA molecules for the effective suppression (silencing) of murine IL-4 gene in vitro. Six potentially effective variants of siRNA molecules were designed in silico and their ability to suppress the expression and production of the target gene in vitro was estimated. It was demonstrated that two of these siRNAs variants were able to significantly suppress both IL-4 expression and production.

Interfering RNA (RNAi) is a powerful tool to silence gene expression on the level of mRNA. To knock-down gene expression by using RNAi two major methods of mRNA silencing exist. First method utilizes siRNA (small interfering RNA), a readily processed dsRNA, that enters RISC complex and destroy target mRNA after transfection into the cells. The second method based on the construction of plasmid DNA that expresses shRNA (short harpin RNA) from U6 or CMV promoter. shRNA gets processed by Drosha and Dicer RNAses inside the cell before it translocates to the cell cytoplasm and affects the level of target RNA. In this study we modified lentiviral vector pGIPZ expressing tFP-IRES-Puro-shRNA(mir30) cassette by introducing BamH I restriction site downstream of this cassette. This modification makes possible to clone specific shRNA sequences in pGIPZ vector using XhoI/BamHI restriction sites instead of the original recombination. Three shRNAs against phosphoprotein P of respiratory sinthitial virus (RSV) and shRNA against human CD43 as a control were generated and cloned into modified so-called pCIPD vector. Monkey kidney cells MA-104 were stably transduced with four shRNA constructs. In conclusion, the generated lentiviral vector pCIPD can be successfully used for efficient gene silencing and virus replication in a broad variety of cells.

Two fragments of conjugative plasmid p19 (95 kb) from the Bacillus subtilis 19 soil strain were cloned and sequenced; these fragments carry genes, products of which are indispensable for the conjugative transfer. One of the fragments 4518 bp in size carries five open reading frames and their fragments (ORF1–ORF5). The protein corresponding to ORF4 is homologous to proteins from the family VirD4. Inactivation of ORF4 and ORF1 by insertional mutagenesis caused a three-to-fivefold decrease in the frequency of plasmid p19 conjugative transfer. Another 2932-bp fragment of p19 was shown to possess a rep region homologous to the rep region of plasmid pBS72 from the B. subtilis 72 soil strain and a novel ORF (ORF6); the protein corresponding to this ORF contains the HTH motif typical for DNA-binding proteins.