RU2480219C1 - Method of treating pox - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science and medicine. What is presented is the use as an antivariolic agent of a soapy amphiphilic high-polymer RNA of Saccharomyces cerevisiae recovered from dry baker's yeast with alkali titrated oleic acid or sodium dodecyl sulphate by treating recovered RNA in oleic acid.

EFFECT: invention provides the rapid and effective treatment of the diseases caused by poxviruses, with natural interferon inductor recovered from dry baker's yeast.

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Description

The invention relates to the field of veterinary medicine and medicine, in particular to agents having anti-arterial action, and can be used for the prevention and treatment of diseases caused by poxviruses.

According to WHO, mortality from infections continues to occupy a leading place on the globe, and in fact, infectious diseases claim the lives of about 15 million people annually. The second half of the XX and the beginning of the XXI century are characterized by an increase in the frequency of diseases caused by viruses, as well as by the identification of the viral nature of diseases of previously unknown etiology and the discovery of new viruses pathogenic to humans.

Another aspect of this problem is the emergence of drug resistance in well-known viral agents. This makes it necessary to constantly test modern viral strains for resistance to chemotherapy and requires antiviral therapy with several antiviral drugs with different mechanisms of antiviral action to prevent the formation of drug-resistant viral strains. Therefore, the question of the need to develop and search for new antiviral drugs seems extremely important and especially relevant.

The existence of rodent-related natural foci of at least 6 out of 11 known viruses belonging to the genus Orthopoxvirus; the growing aggravation of the epidemic of monkeypox in equatorial Africa with an increase in mortality among people by an average of 9.8%; the ability to save the virus in the corpses of people buried in the permafrost of Eurasia and America; the threat of bioterrorism due to unaccounted for virus stocks stored elsewhere and in someone else; the lack of vaccine immunity in the population after the termination of vaccination and vaccine production 30 years ago on the recommendation of WHO - all this makes the risk of an exacerbation of the epidemic situation with catastrophic consequences now and in the foreseeable future even higher than 20-30 years ago [1].

Antiviral drugs are likely to be the main tools to minimize the catastrophic consequences of a sudden exacerbation of the epidemic.

In connection with the foregoing, and the fact that the scientific community and health authorities do not have effective prophylactic and therapeutic drugs adapted for mass use against poxviruses, research is needed to find and develop such drugs.

In the development of anti-fire remedies for initial screening of the effectiveness of drugs, including against smallpox virus, other poxviruses can be used, in particular in animal models, the ectromelia virus. At the final stages of evaluating the effect of drugs, confirmation of the preliminary results obtained requires the use of live variola virus.

The aim of the present invention is a quick and effective treatment of diseases caused by poxviruses with a simple, low-cost preparation in production.

The goal is achieved in that a natural highly effective interferon inducer is used as an anti-foaming agent, extracted from dry baker's yeast using alkali-titrated oleic acid or sodium dodecyl sulfate by treating the isolated RNA with oleic acid - a soapy amphiphilic complex of high-polymer Saccharomyces cerevisia rna with market name - Vitalang-2). It seems relevant to study its effect on the ectromelia virus in laboratory animal models.

An example implementation of the proposed method

Animals

We used healthy sexually mature animals — outbred white mice of both sexes weighing 14–16 g, obtained from the laboratory animal nursery of the Federal State Institution of Science and Science of the World Bank “Vector”. All experiments were carried out in accordance with [2].

168 mice were used in the experiment, of which 126 animals were used in the experiment (18 groups of 7 mice per group) and 42 in the control (6 groups of 7 mice in the group).

The drug Vitalang-2

Vitalang-2 was obtained according to the method [3], sterilized according to the method [4], dissolved in sterile physiological saline (RF), shaking for 15-30 minutes on the day of the experiment.

Viruses

We used the ectromelia virus, strain K-1, obtained from the department of the collection of microorganisms of the Federal State Institution of Science and Science of the Scientific Center "Vector" and developed on the transplanted Vero cell line (green monkey kidney cells) with a titer of 9.33 × 10 ⁵ PFU / ml.

Testing the antiviral activity of Vitalang-2 (prophylactic regimen)

Vitalang-2 was administered to mice intramuscularly three times during the day with an interval of 4 hours (11.00, 15.00 and 19.00) in a total dose of 0.1 mg / kg of animal weight (3 groups of 7 mice per group), 0.5 mg / kg masses (3 groups of 7 mice per group) and 1.0 mg / kg mass (3 groups of 7 mice per group) in a volume of 200 μl / single administration. As a negative control, 3 groups of 7 mice were used, which were intramuscularly injected with RF in a volume of 200 μl / one injection three times during the day at the same time as in the experiment. One hour after the third injection of all preparations (20.00), the animals were infected intranasally under ether anesthesia with a lethal dose (10 LD ₅₀ containing 2.57 lg PFU) of ectromelia in a volume of 40 μl / mouse. The death of animals was taken into account daily for the next 16 days, the percentage of death, the coefficient of protection and the average life expectancy were estimated.

Testing the antiviral activity of Vitalang-2 (treatment regimen)

The mice of the experimental and control groups were infected intranasally under ether anesthesia with a lethal dose (10 LD ₅₀ containing 2.57 lg PFU) of ectromelia virus in a volume of 40 μl / mouse (10.00).

After 1 h (11.00) after infection, the mice were injected with Vitalang-2 intramuscularly and then twice more during the day with an interval of 4 hours (15.00 and 19.00) in a total dose of 0.1 mg / kg of animal weight (3 groups of 7 mice per group), 0.5 mg / kg of weight (3 groups of 7 mice per group) and 1.0 mg / kg of weight (3 groups of 7 mice in the group) in a volume of 200 μl / single administration. As a negative control, we used 3 groups of 7 infected mice, which after 1 h were injected intramuscularly with RF in a volume of 200 μl / one injection and then twice more during the day with an interval of 4 hours (15.00 and 19.00) in the same volume. The death of animals was taken into account daily for the next 16 days, the percentage of death, the coefficient of protection and the average life expectancy were estimated.

Statistical processing

Statistical data processing was performed using the Statistica 6.0 application package using Student t-test, as well as Mann-Whitney U-test. The significance of differences in average values was established using t-student test at p≤0,05.

Research results

Due to the fact that there is currently no commercial antiviral drug for poxviruses, no positive control was used in these experiments, and FR was used as a negative control. A significant advantage in the number of surviving animals compared with the control (19.1% in the prophylactic and therapeutic regimens) was observed both in the groups receiving Vitalang-2 before infection and in the groups receiving this drug after infection with a lethal dose of mice (10 LD ₅₀ ) ectromelia virus. In these experiments, the protective effect depended on the dose of Vitalang-2 (the number of surviving mice was 28.6; 38.1 and 47.6% when using the treatment regimen and 42.9; 52.4 and 61.9% when using the prophylactic schemes at doses of the drug - 0.1; 0.5 and 1.0 mg / kg of animal weight, respectively). The average life expectancy in mice infected with 10 LD ₅₀ ectromelia virus in all groups treated with Vitalang-2 before infection and after infection, with the exception of one group (at a dose of 0.1 mg / kg in the prophylactic regimen), was significantly higher than in control.

Thus, the drug Vitalang-2 is an effective anti-arsenic and after appropriate preclinical and clinical trials it can be recommended for the treatment of diseases caused by poxviruses. Treatment can be started at any stage of the disease, the therapeutic dose is 1 mg / kg of animal or human body weight.

This work was financially supported by the Ministry of Education and Science of the Russian Federation (State contract No. 16.512.11.2018 of February 11, 2011).

Information sources

1. Lvov D.K., Zverev V.V., Gunzburg A.L., Marennikova S.S., Paltsev M.A. Smallpox - dormant volcano // Issues of Virology. - 2008. - No. 4. - S. 4-8.

2. European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes. Strasbourg, 1986.

3. Pat. 2403288 RF. The method of obtaining high-polymer RNA from dry baker's yeast / Yamkova T.V., Zagrebelny S.N., Panin L.E. Yamkova V.I .; publ. 11/10/2010, Bull. No. 31.

4. Pat. 2397988 RF. The method of terminal sterilization of high-polymer yeast RNA / Yamkova T.V., Kuzovkova E.V., Zheleznova Yu.P., Zagrebelny S.N., Panin L.E. Yamkova V.I .; publ. 08/27/2010, Bull. Number 24.

Claims (1)

Use as an anti-fire remedy for the treatment of diseases caused by poxviruses with soap of the amphiphilic high-polymer RNA Saccharomyces cerevisiae extracted from dry baker's yeast using alkali-titrated oleic acid or with sodium dodecyl sulfate by treating the isolated RNA with oleic acid.