[Development of drugs on the basis of high-polymeric double-stranded RNA for antiviral and antitumor therapy]. / Sozdanie lekarstvennykh preparatov na osnove vysokopolimernykh dvuspiral'nykh RNK dlia protivovirusnoi i protivoopukholevoi terapii.

The review summarizes literature data on the development of drugs based on natural and synthetic high-polymeric double-stranded RNA, and their antiviral, immunoadjuvant and antitumor properties. Special attention is paid to cell receptors responding to exogenous dsRNA, the paths of dsRNA-dependent antiviral reaction, ability of dsRNA to inhibit growth and induce apoptosis ofmalignant cells. It has been shown that enhancing the innate immune response with dsRNA can be an effective component in improving methods for treating and preventing infectious and cancer diseases. The further use of dsRNA for the correction of pathological processes of different origin is discussed.

Development of Drugs Based on High-Polymeric Double-Stranded RNA for Antiviral and Antitumor Therapy.

Abstract-The review summarizes literature data on the development of drugs based on natural and synthetic high-polymeric double-stranded RNA (dsRNA), their antiviral, immunoadjuvant, and antitumor properties. Special attention is paid to cell receptors responding to exogenous dsRNA, pathways of dsRNA-dependent antiviral reaction, ability of dsRNA to inhibit growth and induce apoptosis of malignant cells. It has been shown that enhancing the innate immune response with dsRNA can be an effective component in improving methods for treating and preventing infectious and cancer diseases. The further use of dsRNA for the correction of pathological processes of different origin is discussed.

[An antitumor osteotropic agent based on tumor necrosis factor].

A novel drug for treatment of bone metastases based on human recombinant tumor necrosis factor (TNF-alpha) has been designed. The drug is a molecular structure containing yeast double-stranded ribonucleic acid (dsRNA) covered by the conjugate of polyanion dextran with TNF-alpha and bisphosphonate, alendronic acid. The structure is characterized by the combination of substances possessing antitumor activity (TNF-alpha, dsRNA) and a vector molecule (bisphosphonate) providing tropism to hydroxyapatite, the main mineral component of the bone tissue matrix. The conjugation conditions were optimized and the conjugates of TNF-alpha and alendronic acid with dextran were synthesized. Molecular structures were obtained by self-assembly, and the resulting complexes were separated by gel filtration on Sepharose CL-6B. The electrophoretic analysis method revealed decreased mobility of dsRNA in the complex with the conjugate as compared to the mobility of the original dsRNA. This confirms formation of the designed structures. Transmission electron microscopy confirmed the presence of particles with sizes of 30-40 nm in the drug. Evaluation by the sorption/desorption method showed a higher affinity of TNF-alpha conjugates to hydroxyapatite as compared to the original TNF-alpha molecules (from 1.0 to 1.8 mol/L vs. 0.3 mol/L of potassium phosphate buffer for desorption, respectively).

[Efficacy of combined administration of ridostin and oseltamivir (tamiflu) for therapy and prophylaxis of experimental infection induced by influenza virus A (H5N1) in mice].

AIM: Study of possibility of treatment-prophylaxis effect increase during combined administration of ridostin and tamiflu in experiments in mice infected with highly pathogenic influenza virus strain A/chicken/Kurgan/05/2005 (H5N1). MATERIALS AND METHODS: Balb/c line mice infected intranasally with influenza virus at 100 and 10 LD50 doses received ridostin and tamiflu as monopreparation or the combined variant before or after the infection. The mice were observed for 16 days, lethality rate, protection coefficient and average life span were evaluated. Virus concentration in lungs was determined by using titration in MDCK cell line. RESULTS: Combined administration ofridostin and tamiflu after the infection increased survivability of the animals when compared with the control group, and reduced influenza virus concentration in lungs. CONCLUSION: Treatment effect during combined administration of ridostin and tamiflu after influenza virus infection increased.

[Biological properties of a prolonged action interferon inductor].

AIM: Evaluation of composite formulation of yeast double stranded RNA with polyglucinum (dsRNA-PG) effect on non-specific antiviral resistance factors in mice in comparison with commercial formulation Ridostin. MATERIALS AND METHODS: dsRNA and Ridostin formulations were injected intramuscularly once at the dose of 5 mg/ml, polyglucinum--at the dose of 3.75 mg/ml. 3, 5, 24, 48 and 72 hours after the injection serum interferon levels, neutrophil oxidation-reduction activity parameters, peritoneal macrophage phagocyte activity levels were analyzed in mice blood samples. RESULTS: New dsRNA and polyglucinum containing composite formulation is a non-specific resistance system stimulator. dsRNA-PG effect on interferon synthesis and mice phagocyte activity was higher than with Ridostin and developed earlier. Neutrophil function activation by the formulation had a prolonged effect. A possible explanation for increased activity of dsRNA and polyglucinum composite formulation is a modulating effect by the polysaccharide component. CONCLUSION: The new formulation may have a more intensive and prolonged protective effect against influenza virus in comparison with Ridostin.

[Antitumor properties of a liposomal form of a differentiation factor HLDF fragment in cultured cells].

Cytotoxic properties of a liposomal form of the HLDF6 hexapeptide, representing an HL-60 cell differentiation factor fragment, have been studied on a murine primary lymphosarcoma cell culture. It is established that the liposomal HLDF6 peptide is capable of inhibiting proliferation and enhancing death of the cells of both LS and RLS lymphosarcoma strains distinguished by their sensitivity to cytostatic agents. The effect of the preparation is determined by its antiproliferative and apoptogenic actions on the cells. Free HLDF6 peptide showed a lower cytotoxic activity with respect to the tumor cells as compared to the liposomal preparation.

Pharmacokinetics and antitumor effects of the drug containing TNF-α in nanoparticles.

Antitumor activity of TNF-α incorporated in nanoparticles (VLP-TNF-α) and dynamics of its accumulation and elimination from the blood and tumor tissue were studied in ICR mice. The VLP-TNF-α preparation exhibited higher antitumor activity compared to free TNF-α, presumably due to longer circulation of the cytokine in the blood and its more intensive accumulation by tumor tissue.

Antitumor and immunomodulating effects of a fragment of HL-60 cell differentiation factor in mice with lewis pulmonary carcinoma.

A six-member HLDF6 peptide, fragment of HL-60 cell differentiation factor, exhibited antimetastatic and immunomodulating effects.

[Investigation of antitumor activity of the hexamer fraction of HLDF-6 peptide as a differentiation factor in hemoblastoma-bearing mice].

A study of antitumor properties of HLDF-6 was carried out in DBA/2 mice with transplantable ascites lympholeukemia P-388 as well as in tumor cell culture. Immunomodulating characteristics were evaluated. Five-fold administration of HLDF-6 peptide to tumor-bearing mice inhibited tumor growth thus extending survival. As a result metabolic activity decreased which was followed by longer survival of lympholeukemia P-388 cells and enhanced cytological effect of peritoneal macrophages on tumor cells of the same strain.

[Detection of the markers of hepatites B and C and herpesvirus infection during pregnancy].

AIM: Identification of clinico-epidemiologic features of pareneteral hepatites (HB and HC) and herpesvirus infections (cytomegalovirus, CMVand herpes simplexvirus, HSV) duringpregnancy. MATERIALS AND METHODS: Two hundred pregnant women as well as 150 women--blood donors who comprised a control group were tested in Cheboksary (Chuvash Republic). There were no persons vaccinated against HB in both groups. Diagnostics of the HB and HC as well as CMV and HSV infections was performed by ELISA--HBsAg, anti-HBs, anti-HBc IgM and IgG, anti-HCV as well as IgM and IgG to CMV and HSV were determined; PCR was used to detect HBV DNA and HCV RNA. RESULTS: Moderate prevalence of HB and HC markers in pregnant (31% and 3% respectively) and donor women (34% and 2% respectively) as well as widespread prevalence of herpesvirus infections' markers (from 71% to 94.5%) was established. The studied women had no clinical manifestations of HB or HC as well as CMV or HSV infections at the time of the study. The study revealed the following: association between complications of pregnancy and detected markers of HB, HC, and herpesvirus infections according to trimester; detection rate of HBV and HCV markers in combination with CMV and HSV markers in pregnant women; association of pregnancy complications with presence of HB and HC markers with combination of herpesvirus infection markers. CONCLUSION: It was shown that pregnant women with presence of markers of studied infections present a risk group for development of miscarriage threat, inflammatory processes in placenta and amniotic membranes, and untrauterine fetal growth retardation.

[Effects of the dsRNA interferon inducer on the interaction between macrophages and Mycobacterium tuberculosis in vitro]. / Vliianie induktora interferona na osnove dvuspiral'noi RNK n vzaimodeistvie makrofagove i Mycobacterium tuberculosis in vitro.

Effects of the IFN inducer, yeast dsRNA, produced on the interaction between mouse macrophages and phagocytized mycobacteria were experimentally studied in vitro. Mycobacteria were shown to reproduce in macrophages in their initial infection at a ratio of 1:1.25, 1:2.5, 1:5 and 1:10, which was confirmed by an increased insertion of 5.6-[3H]-uracil in M. tuberculosis H37Rv; they also had a destructive impact on macrophages as verified by a higher release of lactic dehydrogenase (LDG) from macrophages. The dsRNA preparation, 40.0, 80.0 and 120.0 microg/ml, was demonstrated to decrease the insertion amount of labeled uracil in phagocytized mycobacteria at a macrophage:mycobacteria ratio of 1:10 and 1:100. The effect depended on a preparation dose and infection degree of macrophages. A decreased release of specific LDG from infected macrophages was shown under the same conditions. The dsRNA affects the interplay of macrophages and phagocytized mycobacteria through inhibiting the vitality of intracellular mycobacteria and through enhancing the stability of macrophages. Special studies denoted that dsRNA, 40.0 microg/ml and 120 microg/ml, activated the production of peroxidation compounds by neutrophils, which phagocytized the sheep erythrocytes. Finally, a possible mechanism of dsRNA impact on the interaction between macrophages and mycobacteria phagocytized by them is under discussion.

[Results of Befnorin clinical trails in healthy volunteers]. / Rezul'taty klinicheskogo ispytaniia preparata benforina na zdorovykh dobrovol'tsakh.

Clinical trails of Befnorin based on the human recombinant TNF-beta elaborated at the Research Design and Technology Institute of Biologically Active Substances, "Vector" State Research Center of Virology and Biotechnology, were carried out on healthy volunteers in compliance with a decision passed by the Committee of Medical and Immunobiological Preparations, Russia's Health Ministry. Single Befnorin doses of 5-10(4) U, 10(5) U, 5-10(5) U, and 10(6) U were administered as intramuscular injections. Clinical, biochemical and immunological parameters were registered for 7 days after a single dose. The drug had an impact on the below immunity indices: Fc-phagocytosis of monocytes, migration index and migration inhibition index. The dose of 10(5) U was proven to be most effective and safe. Supposedly, the drug can be effective in the treatment of herpetic diseases.

[Outlooks for using the interferon inducers in the treatment and prevention of influenza and acute respiratory viral diseases]. / Perspektivy ispol'zovaniia induktorov interferona v lechenii i profilaktike grippa i ORVI.

The methods of treatment by using the stimulators of the non-specific resistance system have been growing even more topical. Drugs based on interferon (INF) and its inducers belong to the group. IFN is of key importance in the regulation of antiviral immunity. The use of IFN inducers can be referred to as the most promising approach to enhancing and activating the body resistance mechanisms. The advantages of IFN inducers before IFN drugs, a wide spectrum of antiviral effects, compatibility with many pharmaceuticals, good tolerability by patients and effectiveness in different administration modes served as a basis for designing a new class of antiviral drugs--IFN inducers. A lot of experimental medical-and-biological research was made to evaluate the efficiency of IFN inducers in influenza. They were shown to possess a pronounced therapeutic-and-preventive action in experiment and clinical trials. Synthetic and natural dsRNA belong to IFN inducers. Multiple research made in cell culture and with experimental animals demonstrated a high interferon-inducing and antiviral activity of double-stranded RNA against influenza virus. A number of dsRNA-based IFN inducers are permitted for clinical use. Data obtained in clinical trials of such inducers are indicative of their pronounced therapeutic-and-clinical effect. Hence, the IFN inducers having different structures and origins can be regarded as promising in the treatment and prevention of influenza and acute respiratory viral diseases.

[The impact of compound of double-stranded RNA and hyaluronic acid on the parameters of non-specific resistance in mice]. / Vliianie kompozitsii dvuspiral'nykh RNK i gialuronovoi kisloty na pokazateli nespetsificheskoi ustoichivosti myshei.

The effect of yeast double-stranded RNA (dsRNA) and of hyaluronic acid (HA) compositions produced on the interferon synthesis, peritoneal phagocytic activity of macrophages and on the hematological parameters were studied in an experiment with white noninbred mice. HA was shown to enhance the dsRNA-induced interferon synthesis, to inhibit the leukopenic reaction and to produce no effect on the phagocytosis-stimulating activity. The data obtained are indicative of that HA is a promising preparation regarding its use within the interferon-inducing compositions based on dsRNA.

[Effect of tumor necrosis factor alpha on the efficiency of anti-rabies vaccination]. / Vliianie faktora nekroza opukholi alpha na éffektivnost' vaktsinatsii protiv beshenstva.

Recombinant tumor necrosis factor-alpha (TNF-alpha) improved the survival of random-bred albino mice vaccinated with antirabies vaccine after infection with rabies CVS strain. The agent dose of 0.1 microgram/animal, injected 1 day postvaccination, was the most effective. Improvement of antiviral resistance of non-vaccinated mice under the effect of TNF-alpha suggests that the effect of this factor on nonspecific resistance factors is one of the probable mechanisms of its modulating effect.

[Study of the antiviral activity, pharmacokinetics and toxic properties of liposomal form of human alpha-2b interferon administered topically]. / Izuchenie protivovirusnoi aktivnosti, farmakokinetiki i toksicheskikh svoistv liposomal'noi formy rekombinantnogo alfa-2b-interferona cheloveka pri naruzhnom primenenii.

Human liposomal recombinant alpha 2b-interferon topically applied to laboratory animals was tested for antiviral activity, pharmacokinetics, and toxicity. The interferon was shown to penetrate through the skin and to circulate in the blood of experimental animals longer than its injectable form, and to exhibit its antiviral activity against genital herpes in guinea pigs. It produced no toxic, skin-irritant, or allergic effects in laboratory animals.

[The activation of phagocytosis by recombinant human tumor necrosis factor-beta]. / Aktivatsiia fagotsitoza rekombinantnym chelovecheskim faktorom nekroza opukholi beta.

The functional activity of phagocytic cells of various types was studied in white non-inbred mice by administering recombinant human tumor necrosis beta (rhTNF-beta). It was shown that rhTNF-beta increased phagocytic activity of the peritoneal exudate, spleen and liver macrophages as well as blood polynuclears. Stimulation of neutrophils was demonstrated in earlier times after administration of the preparation as compared to macrophages (3 h and 24 h, respectively). The duration of the macrophage activation effect and its expression depended on the dose of the preparation and were the most notable when rhTNF-beta was administered in doses of 10(3)-10(5) U/20 g. The addition of reopolyglucin, the polysaccharide filler, didn't remove a stimulatory effect of rhTNF-beta on macrophages, but influenced its dynamics. Multiple administration of the preparation didn't cause the phagocytosis stimulation effect.

[The immunoregulatory properties of recombinant human tumor necrosis factor-beta in mice opposite-reacting to antigen]. / Issledovanie immunoreguliatornykh svoistv rekombinantnogo faktora nekroza opukholi beta cheloveka u oppozitnoreagiruiushchikh na antigen myshei.

The effect of 10(3)-10(5) E/20 g doses of the recombinant factor of human beta tumor necrosis (rFNT-beta) on formation of the immune response and macrophage functional activity was studied in CBA and C57Bl/6 mice that differ in genetically determined level of the immune response to an antigen (sheep erythrocytes). The rFNT-beta was found to cause a modulating effect on the cell and humoral links of the immune response. The effect of the agent depended on the dose and the genotype of the experimental animals. It is suggested that the interlinear differences in the intensity of the humoral immune response in rFNT-beta administration may be connected with the different sensitivity to the agent of the peritoneal macrophages of mice of the used lines.