Method for production of sugar-based agents having antiviral and hepatoprotective action

FIELD: medicine, biotechnology, healthy medical and veterinary preparations.

SUBSTANCE: claimed method includes exposing of aqueous isotonic solution of sugar-containing raw materials such as mixture of glucose and fructose in mass ratio of 1:1 with gamma-irradiation in absorbed dose of 25-40 kGy.

EFFECT: agent of standard composition with high antiviral and hepatoprotective action.

2 tbl, 2 ex

 

The invention relates to medicine and biotechnology and can be used to produce the sugars treatment-and-prophylactic drugs for medical and veterinary purposes.

You know getting biostimulant by separating the antigen from bacterial cells or their metabolic products containing patterns belonging to the group of sugars (patent applications France No. 2399845, publ. 13.06.79, MKI A 61 K 39; EPO No. 0045718, publ. 02.10.82, MKI a 61 K 9/06).

These methods of ethnologica, as they require a high degree of purification of secreted active components and their subsequent binding with a pharmaceutical carrier. This reduces biological activity of the target product.

Also known is a method of obtaining biostimulating funds from sugar-containing raw materials by dissolving the honey in water to a hypertonic concentration (Joiris BTW, the Curative properties of honey and bee venom, M., "Midges", 1956, s-136; Kuzmin K.A. Treatment of bee honey and poison, publishing house of Saratov Univ-TA, 1983).

Target product, obtained in a known manner, has a low biological activity. In addition, it is non-standard, because of the complexity and produces different varieties of honey.

Closest to the claimed technical essence is a way of obtaining drugs, budushego antivirus, hepatoprotective and immunomodulatory activity (Patent RF №2155051, epubl. BI No. 24 - prototype)by fractionation of a solution floral honey crystallization in ice, incubation within 3-30 days at a temperature 4-30°s, respectively, followed by clarification by filtration or centrifugation, freezing and radiation treatment to the absorbed dose of 0.1-100 kGy.

There is a method allows to obtain a biologically active agent with a complex therapeutic properties, but having a non-standard composition due to the use as a raw material of the non-standard multi-component substances, namely - flower honey. Furthermore, the method is sequential and time-consuming.

The objective of the proposed technical solution is the standardization of the target product, while maintaining its biological activity and simplification of the method of its production.

The problem is solved by ionizing radiation absorbed dose is 10 to 40 kGy) on aqueous isotonic solution of sugar containing raw material, which is used as glucose and/or fructose.

The inventive method allows to obtain the substance of the standard composition having a high protivovirusnoi and hepatoprotective activity of glucose and/or fructose. Compared to what procedure prototype, the proposed method is simpler and more accessible.

We have found that the radiolysis products of glucose and fructose or mixtures thereof have protivovirusnoi and hepatoprotective activity in terms of the absorbed dose of gamma radiation 10 to 40 kGy, and found that when used in the method of obtaining compounds having these kinds of biological activity of a mixture of two sugars: glucose and fructose, there is a synergistic effect of increasing this activity.

In the study of the prior art applicants were found similar way, with the above-mentioned differences from the prototype to achieve the same technical result, which is obtained in the present method. This allows to make a conclusion about conformity of the present invention, the criterion of "novelty" and "inventive step".

The inventive method also meets the criterion of "industrial applicability", because it offers the use of known materials and technical means.

The method is illustrated by the following examples.

EXAMPLE 1. Distilled water to prepare isotonic solutions of glucose, fructose, and mixtures thereof. Sample ingredients are taken from the calculation of 0.29 gram-molecules per 1 liter of solution (I. Belova, Karczewski CENTURIES, Rudakova N.A. Sokolova, L. F. manufacturing Technology sterile RA the solutions in terms of pharmacies. M, "Medicine", 1982, p.51). So, based on the molecular weight of anhydrous glucose (fructose) 180,18, the amount of glucose (fructose) in 1 l of isotonic is: 0,h,18=52,25, Therefore, isotonic concentration of glucose, fructose or mixtures thereof) equal to 5.2 wt.%. Here and below we take equal sample sugars in the mixture. Similarly prepared control solutions, including additional sugar xylose. Another control is isotonic honey (prototype).

The prepared solutions are subjected to γ-irradiation dose of 10 kGy and 25 kGy. While the honey solution and the solutions of the mixture of sugars is divided into two parts, one of which left unirradiated (control). The biological activity of drugs is examined for white mice (10-14 individuals in the group). Prepared solutions administered intranasally 50 ál for 10 days, then spend a single seed mice intraperitoneally with 0.2 ml of 25% oil solution of the CCI4. The effect of the drugs is estimated by the survival of mice for 4 days. The results are shown in table 1.

The table shows that:

1) sugar xylose pronounced hepatoprotective effect does not possess;

2) of the tested sugars most potent hepatoprotective effect has fructose (death of mice 20%);

3) effect of fructose increases in the presence of glucose (with adequate isotonic concentration is the mixture of ingredients of the death of the mice just 7.7%);

4) γ-irradiation leads to increased hepatoprotective properties of the obtained products in 2-3 times;

5) irradiated mixed solution of glucose and fructose has a more pronounced hepatoprotective properties than the prototype.

EXAMPLE 2. Subjects and control preparations prepared as in example 1 at the absorbed dose of gamma radiation 40 kGy. They have an antiviral effect on the model experiencing fragments chorioallantois shell (FAO) 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 incubated 1 h at 37°for adsorption is carried virus. Later in the wells make subjects and control preparations in 20-fold dilution (10 holes on the drug) and incubated at 37°C for 3 days, after which determine residual infectious activity (IA) virus titration on chicken embryos. The results are shown in table 2.

As can be seen from table 2, the irradiated solutions of glucose and fructose have antiviral action, the most effective use of their irradiated mixture, which is somewhat superior to the prototype (the drug of honey solution).

As explained above examples, the use of the proposed method in comparison with the prototype allows jet to preserve the biological activity of the target product at a high level, and, in the case of using a mixture of glucose and fructose, even more to increase it. The inventive method provides for the standard description of the target product and is more simple in comparison with the method of the prototype.

20,0
Table 1

Hepatoprotective effects of drugs after priming mice CCI4
Ingredients preparation*Absorbed dose, kGyThe percentage of death:
glucosefructosexylosehoney1 day4 days
+---2550,060,0
-+--2520,020,0
--+-2585,792,9
++--257,77,7
++--1014,321,4
++--020,0
+++-258,38,3
---+042,042,0
---+258,08,0
(control: mice do not protect)100,0-
* Presence(+)or absence(-).

Table 2

Antiviral activity of drugs on the model FHA
Ingredients preparation*Absorbed

a dose of 40 kGy
Residual IA, lg EID50Index reduction IA, lg EID50
glucosefructosexylosehoney
+---+3,0±0,052,5±0,03
-+--+3,3±0,072,2±0,04
--+-+5,0±0,05 0,5±0,02
++--+2,3±0,063,2±0,02
++---4,2±0,061,3±0,03
+++-+2,6±0,052,9±0,07
---+-3,7±0,041,8±0,07
---++2,5±0,053,0±0,01
(control: infected cells not treated with a protective tool)5,5±0,03-
* Presence(+)or absence(-).

The method of obtaining funds on the basis of sugars with antiviral and hepatoprotective activity by exposure to gamma-irradiation at an absorbed dose of 25-40 kGy water isotonic solution of sugar containing raw material, characterized in that as the sugar-containing raw materials, use glucose and fructose in a ratio of 1:1 by weight.



 

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