A method of obtaining a medicinal product, which has antiviral, hepatoprotective and immunostimulatory activity

 

(57) Abstract:

The invention relates to medicine. The substance obtained by the fractionation of a solution floral honey crystallization in ice, incubation for 3 to 30 days at a temperature of 4 - 30oC, respectively, with the following clarification by filtration or centrifugation, freeze to ice. Then put her radiation treatment dose of ionizing radiation of about 0.1 - 100 kGy. The invention allows a 30% increase antiviral activity of the drug. table 4.

The invention relates to medicine, namely to methods for drugs with antiviral, hepatoprotective and immunostimulatory therapeutic activity.

"Vetmedin-M is a complex of biologically active substances derived from natural bee honey. The drug has a corrective effect on the system, nonspecific resistance, increases its resistance to viral infections such as influenza and other acute respiratory infections, herpes, bacterial infections, and it is indicated for diseases that cause abnormal liver function and to reduce the hepatotoxic action Lek is artenum Committee of the Russian Federation under N 97/202/4 and approved for use and commercial production. It is a solution of a light yellow color with a characteristic smell of honey and is intended for intranasal /nose drops/. At the specified route of administration of drug healing properties of honey is completely preserved, while in normal, oral, receiving its therapeutic properties largely lost under the action of acids and enzymes of the gastric juice. "Vetmedin-M contains only natural ingredients of honey without preservatives and synthetic additives. It is, first of all, easily digestible monosaccharides - glucose and fructose in a biologically active form, as well as biologically active acid-formic, malic, citric, acetic, gluconic, glucuronic, and other vitamins, micro - and macronutrients that are part of natural honey. The product does not contain impurities that cause allergic reactions, proteins, pollen and polysaccharides, therefore, the tests showed the practical absence of allergenic properties.

Found that "Vetmedin-M" at the same time possesses antiviral, hepatoprotective, and immunokorrigiruyuschy, and hypolipidemic, and antioxidant effect that can be attributed to natural adaptogens, such Keane, Essentiale, acyclovir, timogen.

A method of obtaining bio-stimulating means by dissolving the honey in water to a hypertonic concentration /Medicinal properties of honey and bee venom. M: "Midges", 1956, S. 135-136, Kuzmin K. A. Treatment of bee honey and poison. Publishing house of Saratov University, 1983/. To improve the biological activity of the honey solution obtained by fractionation and crystallization in ice/application N 5002353 from 23.08.91/ further processing of the product of gamma-irradiation /application N 5064879 from 16.07.92/.

However, the target product obtained by these methods has a low lipid-lowering, antioxidant and hepatoprotective activity.

Closest to the claimed technical essence and the achieved result is a method of obtaining compounds having hypolipidemic, antioxidant and hepatoprotective activity of honey bee /patent RF N 2084232, B. I. N 20, 1997/, consisting in the crystallization of honey in ice, followed by incubation for 3-30 days at a temperature of from 4 to 30oC and clarification by filtration or centrifugation. Further, the radiation treatment product gamma-irradiation.

However, the target Peremoga method of obtaining drugs "Vetmedin-M" is the enhancement of antiviral activity of the target product while maintaining all its curative properties.

This objective is achieved in that the substance "of Vitamedica-M, obtained by fractionation of a solution floral honey crystallization in ice, incubation for 3 to 30 days at a temperature of 4 - 30oC, respectively, followed by clarification by filtration or centrifugation, frozen to ice, and then subjecting it to radiation processing dose of ionizing radiation, a component of 0.1 - 100 kGy.

Antiviral activity of Vitamedica-M, obtained by the claimed method, in experiments on the reproduction of the herpes virus in vitro was comparable to that of the most effective foreign drug acyclovir.

Radiation treatment is used in the known methods in obtaining biologically active solutions of honey, but the first time we have found that pre-freezing of the drug before irradiation enhances existing antiviral properties, without weakening other medicinal properties.

Thus, the proposed technical solution has novelty, inventive step and is industrially applicable.

Examples of the implementation of the proposed method.

the surface of the ice, put a layer of previously stored at room temperature non-crystalline floral honey the obtained organic apiaries and held tight control on all parameters of the standard and in the absence of salts of heavy metals and radionuclides in a weight ratio of 4:1 respectively and stand for 20 hours at -20oC. During this period the dissolution occurs honey in ice caused teplomassoperenosa, with subsequent crystallization.

Crystallized honey solution is divided into portions, which are incubated at temperatures and for periods specified in the table. 1. After incubation produce clarification of the target product by filtration through a cotton-gauze filter, then the substance of Vitamedica-M frozen at a temperature of -14oC to ice and subjected to gamma irradiation with a dose of 25 kGy.

The biological activity of the drug have on white mice in the 9 - to 12 individuals in the group, introducing his oral 1 drop of specimen for 10 days, then the mice intraperitoneally injected with 80 mg/kg geksenala - hypnotic, neutralization which occurs only in the liver. On the biological activity of the target product is judged by the duration geksenalovy sleep mice compared with the duration geksenalovy sleep mice following degausse irradiation after freezing of the drug prior to the formation of ice /prototype/.

Control 2. Geksenal injected mice not protected by any drug.

The test results are given in table. 1.

Example 2. A solution of honey get as in example 1, with the following values of the operational parameters of the stage of incubation: duration - 15 days, the temperature of the 22oC. the Yield of the target product is 95%. The radiation dose was 25 kGy, the temperature of the freezing substance of Vitamedica-M" before gamma-irradiation -20oC.

Biological /antioxidant activity of drugs tested on rats male weighing 240 - 260 g /8 - 10 individuals in the group/. Rats get hypercholesterinemia diet /GHD/: 2.5% cholesterol, 0,12% 6-thiuragyl and 30% mixture of fats /heated sunflower oil and lard in a ratio of 1: 1/. The test drug is administered intranasally in a dilution of 1:10 to 1 drop in for 24 days. About the antioxidant activity of the target product is judged by the content of lipid hydroperoxides in plasma.

Control 1 - prototype. Control 2 - unprotected rats.

The test results are given in table. 2.

Example 3. A solution of honey get as in example 2, and at the end of the incubation solution of honey lighten centrifuging the ice at the temperature of -10oC, the absorbed dose of gamma irradiation amount is 0.1 kGy and 25 kGy; 100 kGy.

Test hepatoprotective action of the drug on white mice /9 - 12 individuals in the group/. It is introduced in a dilution of 1:100 route 50 ál for 10 days, then spend a single seed of mice against carbon tetrachloride /CCl4/ intraperitoneally with 0.2 ml of 25% oil solution CCl4. The effect of the drug is estimated by the survival of mice for 4 days.

Control 1. Mice protects irradiated product of the same series, but without pre-freezing /prototype/.

Control 2. CCl4enter unprotected mice.

The results are shown in table. 3.

Example 4. A solution of honey get as in example 3. Portions of the obtained preparation is frozen to ice at a temperature of -10oC, the absorbed dose of gamma irradiation amount is 0.1 kGy, 25 kGy.

Test antiviral activity carried out on the model experiencing fragments chorioallantois shell /FHEO/ chick embryos. For this influenza virus A/Hong Kong/1/68/H3N2/ infectious activity of 100 EID50/0.2 ml contribute to the wells planchettes with FAO in culture medium 199 and irusa titration on chicken embryos. The results are shown in table. 4.

Control 1. Infected cells treated with bioactive honey solution of this party, is not subjected to freezing before irradiation /prototype/.

Control 2. Infected cells not treated with a protective agent.

Example 4. Preparation and testing of the preparations was carried out as in example 4, but was pre-irradiated frozen at a temperature of -10oC substance "Vitamedica-M" flow accelerated electrons produced at the accelerator "Elite 2" /pulse width 3 μs, the electron energy of 1.5 MeV/t and the exposure dose was 100 kGy. The results are shown in table. 4.

Radiation treatment of substance "Vitamedica-M doses more than 100 kGy not economically viable and that doses less than 0.1 kGy does not lead to a noticeable positive effect from exposure.

As can be seen from the table. 1, 2, 3 biological activity obtained by the claimed method of the medicinal product "Vetmedin-M" on a number of indicators stored at the level of performance of the preparation obtained by the method of the prototype.

As can be seen from the table. 4 antiviral activity of the drug "Vetmedin-M received the claimed enables you to get the drug "Vetmedin-M with higher antiviral activity while saving the most complex therapeutic properties of the drug.

The method of obtaining drugs by fractionation of a solution floral honey crystallization in ice, incubation for 3 to 30 days at a temperature of 4 - 30oaccordingly, with the subsequent clarification by filtration or centrifugation, characterized in that obtained after clarification of the substance is frozen to ice, and then subjecting it to radiation processing dose of ionizing radiation, a component of 0.1 - 100 KGy.

 

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