RU2445108C1 - Preparation showing bactericidal and antimetastatic activity - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: preparation shows high bactericidal and antimetastatic activity that is ensured by the presence of a water extract of shelf fungus and birch bark taken in weight proportions 1:4.

EFFECT: preparation shows bactericidal and antimetastatic activity.

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Description

The invention relates to medicine, in particular to the field of pharmacology and pathophysiology, and can be used as an agent with anti-inflammatory and immunostimulating effects.

It is known to use decoctions of birch buds and leaves as an anti-inflammatory agent for colds, as well as in the treatment of tuberculosis (V.P. Makhlayuk. Medicinal plants in folk medicine. - Saratov: Volga Book Publishing House, 1993 - P.58) .

However, this decoction has a weak bactericidal effect.

A known composition of ingredients with biological activity (options) containing the culture fluid of Kombucha and the infusion of Chaga (RF patent No. 2280394, publ. 16.12.04.).

However, this drug has a weak antimetastatic activity and is intended for use as a food additive in the production of various kinds of drinks.

The objective of the invention is to develop a drug with increased bactericidal and antimetastatic activity.

This problem is solved in that the preparation having bactericidal and antimetastatic activity based on an aqueous extract of plant material contains an aqueous extract (extract) of chaga and birch bark, taken in a weight ratio of 1: 4.

The invention explains the description of the drug and the results of laboratory studies of its bactericidal and antimetastic properties.

The drug is obtained according to well-known technology, using any type of installation to obtain aqueous extracts (extracts) from plant materials. As the latter, chaga and birch bark, taken in a weight ratio of 1: 4, are used. The obtained extract (preparation) is a light brown liquid, which can be used both directly - with external, and in the form of aqueous and alcoholic solutions of the required concentration - for internal use.

Determination of the bactericidal activity of the drug was carried out by the method of vertical diffusion on a solid nutrient medium "NEW" (G. G. Mordovskaya, 1972) using a laboratory strain of MBT H37Rv.

During the study, a sample of the culture of the laboratory strain H37Rv was placed in a sterile porcelain mortar, ground to a homogeneous mass, gradually adding sterile saline. The resulting culture suspension was standardized according to the bacterial turbidity standard corresponding to 1 billion microbial bodies (10 units of GKI). 0.1 ml suspension of MBT was seeded in test tubes with nutrient medium, after which they were placed in a horizontal position in a thermostat for 24 hours at a temperature of 37 ° C.

Were prepared aqueous solutions of the study drug in a concentration of: 100; fifty; 25; 12.5; 6.25%.

The test preparation of the appropriate concentration was added to each tube seeded with the H37Rv strain and placed in a thermostat at a temperature of 37 ° C.

The effect of the drug of each concentration on the office was studied in three parallel rows. For comparison, isoniazid in similar concentrations was used as a control. The results were recorded after 10 days, taking into account the size of the zones of growth inhibition of the office (in mm). The results are presented in the table.

Table 1 The results of the study of the bactericidal properties of the drug Preparations The concentration of the drug,% one hundred fifty 25 12.5 6.25 The studied drug (Water extract of chaga and birch bark) 58 fifty 40 35 27 55 fifty 40 thirty 25 52 46 43 thirty 27 55 ± 2 48.7 ± 1.8 41 ± 1.3 31.7 ± 2.2 26.3 ± 0.9 The concentration of isoniazid, mcg / ml Isoniazid 25.0 12.5 6.25 3.12 1,5 > 70 60 60 45 40 > 70 58 60 40 40 > 70 55 45 40 37 > 70 57.7 ± 1.8 55 ± 6.7 45 ± 3.3 39 ± 1.3

From the data presented in table 1 it follows that according to its bactericidal properties, the proposed preparation (water extract (extract) of chaga and birch bark), including at a 50% concentration, corresponds to the bactericidal properties of isoniazid at a concentration of 25.0 μg / ml.

To determine the antimetastatic activity of the drug, experiments were carried out on mice with subcutaneously transplanted tumors that are part of the panel of tumor strains for screening ICBM antitumor activity: breast adenocarcinoma Ca755, lymphocytic leukemia P388 and melanoma B16. Tumor strains were introduced in the volume: P388 - 1 million cells / mouse, Ca755 and B16 - 40 mg of tumor tissue per mouse in 0.3-4 ml of culture medium 199.

In the experiment with Ca755, from 6 groups of 8 mice, 5 groups received treatment with the proposed drug, and one group, the control group, was injected with saline. The drug was used in 100% and 20% concentrations after dilution with sterile saline by introducing into the abdominal cavity in a single dose of 0.1-0.5-1.0 ml per mouse. The introduction of the drug began 72 hours after transplantation of the tumor strain and was carried out daily for 5 days. The total dose of the drug was 0.5-2.5-5.0 ml / mouse, respectively.

In the experiment with P388, there were 2 groups of 10 mice: one group was injected with a preparation of 20% concentration obtained as a result of its dilution with physiological saline, and the other, control, physiological saline. The drug was administered into the abdominal cavity at a single dose of 0.5 ml / mouse and was carried out daily for 5 days, starting 72 hours after transplantation of the tumor strain. The total dose was 2.5 ml / mouse.

In the experiment with B16, out of 3 groups of mice, two groups received a preparation of 20% concentration and one, control, was injected with saline. The drug was administered into the abdominal cavity daily also for 5 days, starting 72 hours after transplantation of the tumor strain. Single doses were 0.25 and 0.5 ml per mouse; total doses, respectively, 1.25 and 2.5 ml / mouse.

After the end of the experiment, the tumors were measured, the average volumes were determined, and the standard efficiency indicator, tumor growth inhibition (SRW), was calculated, which was expressed as a percentage of the control group. In the assessment, we proceeded from the fact that the effectiveness indicator in the initial study (screening) was TPO≥50%. The reliability of the result was evaluated by the standard student method with the calculation of the confidence interval in the compared groups. The differences were considered significant at p≤0.05.

When analyzing the results of the experiments, the following is established.

From experience with Ca755. In the group of mice that were injected with the drug 100% concentration, all mice died during the first day after the second injection. An injected drug was found in the abdominal cavity of fallen mice that was not absorbed into the body. The cause of death is the local irritating effect of the administered drug on the wall of the abdominal cavity. In the groups receiving the diluted preparation of 20% concentration, its tolerance was good. A reliable antitumor effect of TPO = 68-69-72% (the last value is reliable, p <0.05), corresponding to the minimum criterion, was obtained with the introduction of a 20% solution in a single dose of 0.5 ml per mouse, the total dose of 2.5 ml / mouse. Summary data are presented in table 2.

table 2 Antimetastatic efficacy of the drug in relation to Ca755 Group n = 10 Single dose (ml / mouse), mode of administration Day after tumor inoculation 10 fourteen eighteen Vcp, mm ³ TPO% Vcp, mm ³ TPO% Vcp, mm ³ TPO% Control, nat. rr 0.5 × 5 times 1070 ± 600 * - 6308 ± 3946 - 12673 ± 5172 - Preparation (Water extract of chaga and birch bark, 100% concentration) 1.0 × 2 times 10 mice fell from toxicity on day 2 0.5 × 2 times 9 out of 10 mice fell from toxicity on day 2, in one surviving mouse the size did not differ from the control 0.1 × 5 times 1001 ± 714 6 2517 ± 1577 60 5252 ± 4403 54 Preparation (Water extract of chaga and birch bark, 20% concentration) 1.0 × 5 times 546 ± 374 49 35639 ± 2843 42 5425 ± 4123 57 0.5 × 5 times 339 b scatter 68 1918 ± 1477 69 3369 ± 2697 72 ** Notes: all solutions were administered intraperitoneally daily for 3-7 days after tumor transplantation;
SRW was determined on the 3rd, 7th and 11th day after the end of treatment; * - confidence interval; ** - p <0.05

In the experiment with B16, a group of mice that were injected with the drug at 20% concentration, tolerated it well. A long-term reliable antitumor effect corresponding to the minimum criterion was noted, TPO = 95-96-94% (p <0.05) on days 2-6-12 after the end of the experiment. Summary data are presented in table 3.

Table 3 Antimetastatic efficacy of the drug in relation to strain B16 Group n-10 Single dose (ml / mouse), mode of administration Day after tumor inoculation 9 13 19 Vcp, mm ³ TPO% Vcp, mm ³ TPO% Vcp, mm ³ TPO% Control, nat. rr - 601 ± 174 * - 2137 ± 682 - 6299 ± 1862 - Preparation (Water extract of chaga and birch bark 20% concentration) 0.5 × 5 times 63 ± 56 90 ** 246 ± 682 88 ** 938 ± 429 85 * 0.25 × 5 times 32 ± 31 95 ** 74 ± 63 96 ** 409 ± 283 94 ** Notes: all solutions were administered intraperitoneally daily for 3-7 days after tumor transplantation;
SRW was determined on the 2nd, 6th and 12th day after the end of treatment; * - confidence interval; ** - p <0.05

In the experiment with P388, in the group of mice that received the drug at 20% concentration, a significant short-term antitumor effect was observed on day 4 of TPO = 52% (p <0.05), which corresponds to the minimum criterion.

In general, studies have shown that the proposed drug has a high bactericidal and antimetastatic activity.

Claims (1)

A drug with bactericidal and antimetastatic activity based on an aqueous extract of plant materials, characterized in that it contains an aqueous extract (extract) of chaga and birch bark, taken in a weight ratio of 1: 4.