RU2454234C1 - 1,3-diethylbenzimidazolium salts as immunostimulating drugs and based pharmaceutical compositions - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to a pharmaceutical composition and to a drug of 1,3-diethylbenzimizadolium salts of general formula

: wherein X^- = anion.

EFFECT: there are prepared new compositions on the basis of salts of said formula showing immunostimulating action.

2 cl, 2 tbl, 6 ex

Description

FIELD OF TECHNOLOGY

The invention relates to the field of immunology, in particular to means of stimulating the immune system.

BACKGROUND

It is known that the drug Procodazole - a benzimidazole derivative (1H-benzimidazole-2-propanoic acid) is a non-specific active immunoprotective agent (Patent RU 2197964, publ. 02.10.2003).

The disadvantage of this drug is that it is effective only in combination with other benzimidazole drugs in the treatment of viral infections.

Closest to the proposed is the drug dibazole, used as a mild immunostimulating drug (Mashkovsky MD 2003. Medicines. M: Novaya Volna, T.1, p. 398-399), also a benzimidazole derivative - 2 -benzylbenzimidazole hydrochloride. However, dibazole has insufficient immunostimulating activity. In addition, dibazole is not a salt of the benzimidazolium cation, which has high pharmacological activity.

SUMMARY OF THE INVENTION

The objective of the invention, which determines its purpose, is the creation of new drugs, as well as their acceptable pharmaceutical compositions having a pronounced immunostimulating effect, with high therapeutic efficacy, while reducing unwanted side effects, reducing the cost of treatment.

The technical result of the claimed invention, which can be achieved by its implementation, is the creation of a group of compounds, as well as their acceptable pharmaceutical compositions with high activity in the treatment of immunodeficiency diseases.

The technical result is achieved with substances active in the treatment of immunodeficiency diseases from the group of 1,3-diethylbenzimidazolium salts of the general formula:

where X ^- = anion.

The technical result is also achieved by a pharmaceutical composition comprising a 1,3-diethylbenzimidazolium salt of the above general formula and at least one pharmaceutically acceptable excipient.

The following substances may be used as pharmaceutically acceptable excipients in a composition having immunostimulating activity, including a salt of 1,3-diethylbenzimidazolium:

for tablets, potato starch, or sodium stearate, or talc, or other excipients, or various combinations thereof;

for capsules - gelatin (shell), potato starch, or sodium stearate, or talc, or other excipients, or various combinations thereof;

for injection form - saline or water for injection;

for ointments - polyethylene glycol 400, or polyethylene glycol 1500, or low molecular weight polyvinylpyrrolidone, or other excipients, or various combinations thereof;

for suppositories - polyethylene glycol 400, or polyethylene glycol 1500, or low molecular weight polyvinylpyrrolidone, or other excipients, or various combinations thereof;

for drops for oral administration and topical use, ethyl alcohol, or potassium iodide, or low molecular weight polyvinylpyrrolidone, or other excipients, or various combinations thereof.

A method of obtaining salts of 1,3-diethylbenzimidazolium and their acceptable pharmaceutical compositions for the treatment of immunodeficiency diseases is safe for individuals involved in the process.

MODES FOR CARRYING OUT THE INVENTION

The proposed funds are obtained as follows.

Example 1. 1,3-diethylbenzimidazolium chloride

59 g (0.5 M) of benzimidazole and 68 g (0.5 M) of sodium trihydrate are placed in 150 ml of o-xylene, 64.5 g (1 M) of ethyl chloride are added and the reaction mixture is boiled for 3 hours. It is cooled on ice, the obtained crystalline product is washed with ice water, filtered and dried for 24 hours at + 55 ° C. Fine crystals from white to light yellow. Yield 72%. The structure of the obtained compound is confirmed by elemental analysis. Found: N 13.4%, Cl 16.8%. Calculated: N 13.3%, Cl 16.9%.

Example 2. 1,3-diethylbenzimidazolium iodide

59 g (0.5 M) of benzimidazole and 68 g (0.5 M) of sodium trihydrate are placed in 150 ml of o-xylene, 156 g (1 M) of ethyl iodide are added and the reaction mixture is boiled for 3 hours. It is cooled on ice, the obtained crystalline product is washed with ice water, filtered and dried for 24 hours at + 55 ° C. Small crystals from white to light yellow or light pink. Yield 68%. The structure of the obtained compound is confirmed by elemental analysis. Found: N, 9.2%; J, 43.7%. Calculated: N 9.3%, J 42.0%.

Example 3. 1,3-diethylbenzimidazolium sulfate

59 g (0.5 M) of benzimidazole and 20 g (0.5 M) of sodium hydroxide are placed in 300 ml of ethyl alcohol, 77 g (0.5 M) of diethyl sulfate are added and the temperature of the reaction mixture is maintained for 3 hours in the range of +50 -60 ° C. It is cooled at -20 ° C, the obtained crystalline product is washed with ice water, filtered and dried for 24 hours at + 55 ° C. Small crystals of white color. Yield 65%. The structure of the obtained compound is confirmed by elemental analysis. Found: N 12.4%, S 7.1%. Calculated: N 12.6%, S 7.2%.

Pharmaceutical compositions are prepared as follows.

Example 1

Fine crystals of 1,3-diethylbenzimidazolium chloride (0.4 g) are mixed with potato starch (9.6 g) and 10 mg tablets are formed under oral pressure for oral administration.

Example 2

Finely ground crystals of 1,3-diethylbenzimidazolium iodide (0.4 g) are mixed with potato starch (9.6 g) and 10 mg tablets are formed under oral pressure for oral administration.

Example 3

Fine crystals of 1,3-diethylbenzimidazolium sulfate (200 mg) were dissolved in physiological saline (10 ml), the resulting 2% injection was sealed in ampoules.

The pharmacological efficacy of the obtained 1,3-diethylbenzimidazolium salts was studied in white mice. For in vivo studies, the influenza virus A / Aichi / 2/68 (H3N2) was used, which was previously adapted for propagation in the lungs of mice at a dose of 100 LD ₅₀ . Infectious activity of the virus was determined by the method of Reed and Mench upon infection of mice with ten-fold dilutions of the suspension in the lungs.

The ability of the drug to have an immunostimulating effect was judged by changes in the immune status of mice infected with adapted influenza virus in the experimental groups compared with the group of animals that did not receive the drug.

5 groups of animals were formed, 10 mice in each group.

Animals of the 1st group were not exposed to any effects and they constituted a group of intact animals.

Animals of the 2nd group were infected, but did not receive drugs.

Used an infectious dose of the virus, causing 60-70% mortality in mice.

The drug composition of 1,3-diethylbenzimidazolium chloride was administered orally to animals of the 3rd group, 1 tablet (corresponding to a dose of 20 mg / kg) 2 times a day 24 hours before the infection of mice with the virus.

The drug composition of 1,3-diethylbenzimidazolium iodide was administered orally to animals of the 4th group, 1 tablet (corresponding to a dose of 20 mg / kg) 2 times a day 24 hours before the infection of the mice with the virus.

Animals of the 5th group were injected orally in the form of a 2% solution of 1,3-diethylbenzimidazolium sulfate 0.2 ml (corresponding to a dose of 20 mg / kg) 2 times a day 24 hours before infection of mice with the virus

The animals were observed for 20 days after infection, taking into account the death of mice from influenza in the groups of treated animals and in the control group (mice that did not receive the drug).

To study the pharmacological activity of the drug, the blood and lungs of 5 mice were taken 6 and 12 days after infection.

Analysis of the results of the experiments was carried out according to the indicators of the immune system of infected animals in groups receiving the drug in relation to the control group.

The characterization of the immune status was carried out using a complex of methods, allowing to evaluate both cellular and humoral parts of the immune system.

The average values for each group of animals with the average deviation in the Microcoft Excel system were calculated.

Digital material was subjected to statistical processing using the student method.

The study of drugs against the background of infection of animals with the influenza virus revealed their distinct participation in the formation of the body's immune response to infection. It should be noted that salts of 1,3-diethylbenzimidazolium had a protective effect against experimental influenza infection in all regimens of drug administration.

The revealed therapeutic effect of the drugs proceeded against the background of a statistically significant decrease in the titer of influenza virus in the lungs of animals relative to the control, which was noted on the 6th and especially on the 12th day of the experiment (tables 1,2). That is, it must be concluded that the salts of 1,3-diethylbenzimidazolium have protective activity against the influenza virus A / Aichi / 1/68 (H3N2).

Data on the protective effect of the drug are explained by the results of studying the immune status of animals. So, on the 6th day after infection of animals in mice treated with 1,3-diethylbenzimidazolium salts, the blood level of IgM immunoglobulins, which are associated with the primary immune response, was statistically significantly higher than their concentration in untreated animals (Table 1).

In addition, there was a significant excess on the 6th and especially on the 12th day of the experiment in mice of the experimental groups in comparison with control animals the content of IgG immunoglobulins in the blood (Tables 1, 2), which play a major role in ensuring long-term humoral immunity in infectious diseases.

Table 1 Indicators of the immune system of mice 6 days after infection of animals with the virus and drug administration Immune status indicator Intact mice Control group: virus without treatment Experimental groups: the use of salts of 1,3-diethylbenzimidazolium chloride iodide sulfate Monocytes 1.4 ± 0.5 1.2 ± 0.4 3.0 ± 0.7 * 4.4 ± 1.0 * 3.5 ± 0.2 * T-lymphocytes - CD3 +,% 63.8 ± 3.6 45.8 ± 1.7 63.7 ± 1.2 * 60.7 ± 2.3 * 55.6 ± 1.1 * B-lymphocytes - CD20 +,% 14.2 ± 1.33 11.8 ± 2.0 15.4 ± 1.2 * 14.4 ± 1.0 * 15.1 ± 1.4 * T-Helpers / Inductors - CD4 + 52.2 ± 3.92 29.6 ± 1.9 48.8 ± 1.7 * 49.1 ± 1.7 * 42.8 ± 5.4 * IgM immunoglobulins, g / l 0.5 ± 0.4 2.2 ± 0.8 4.2 ± 0.8 * 3.8 ± 0.7 * 4.7 ± 0.6 * IgG immunoglobulins total, g / l 3.0 ± 0.9 4.2 ± 0.8 5.6 ± 0.5 5.0 ± 0.6 5.4 ± 0.8 Infectious virus titer in the lungs (lg) - 2.0 ± 0.7 1.0 ± 0.8 * 1.0 ± 0.5 * 0.8 ± 0.4 *

table 2 Indicators of the immune system of mice 12 days after infection of animals with the virus and drug administration Immune status indicator Intact mice Control group: virus without treatment Experimental groups: the use of salts of 1,3-diethylbenzimidazolium chloride iodide sulfate Monocytes 1.4 ± 0.5 2.2 ± 0.4 1.2 ± 0.1 2.6 ± 0.5 1.3 ± 0.5 T-lymphocytes - CD3 +,% 63.8 ± 3.6 59.2 ± 3.2 67.2 ± 2.4 * 67.8 ± 1.0 * 63.8 ± 4.2 B-lymphocytes - CD20 +,% 14.2 ± 1.33 14.8 ± 1.3 15.1 ± 1.2 14.6 ± 0.8 14.7 ± 1.3 T-Helpers / Inductors - CD4 + 52.2 ± 3.92 47.2 ± 2.8 56.5 ± 3.1 * 56.4 ± 0.5 * 52.0 ± 1.4 IgM immunoglobulins, g / l 0.5 ± 0.4 1.4 ± 0.5 3.5 ± 0.2 * 2.9 ± 0.8 * 3.6 ± 1.1 * IgG immunoglobulins total, g / l 3.0 ± 0.9 5.6 ± 1.4 11.0 ± 1.0 * 10.7 ± 0.7 * 12.3 ± 1.5 * Infectious virus titer in the lungs (lg) - 0.8 ± 0.4 0.2 ± 0.2 * 0.1 ± 0.2 * 0.2 ± 0.1 * * - Significant differences from the control group (p <0.05).

The use of drugs increased the performance of CD4 + markers relative to the control group (Tables 1, 2), which indicates an increase in the number of T-helpers / inductors. These cells are divided into subtypes. Th1 cells activate specific cellular immunity. Th2 cells activate a specific humoral immune response, stimulating the proliferation and differentiation of B cells into antibody-forming cells. The activation of the synthesis of immunoglobulins noted above with the use of drugs can be achieved by increasing the level of T-helpers / inducers of Th2 type.

The effect of drugs on the relative content of monocytes in the peripheral blood was also observed mainly on the 6th day of the experiment (Table 1). An increase in the level of this type of white blood cells, which are the most active phagocytes, was detected relative to the control group.

Thus, the studies showed the undoubted activating effect of 1,3-diethylbenzimidazolium salts on immunity indicators when animals are infected with the influenza virus. A significant difference between the experimental and control results indicates a pronounced immunostimulating effect of pharmaceutical compositions of 1,3-diethylbenzimidazolium salts.

When using the invention, the following conditions are met.

Substances of the claimed structure can be obtained chemically, and their acceptable pharmaceutical compositions can be obtained by technological means. The claimed funds can be used as medicines for the treatment of immunodeficiency diseases in medicine.

Claims (2)

1. The drug having immunostimulating activity, which is a salt of 1,3-diethylbenzimidazolium of the General formula

where X ^- is the anion. 2. A pharmaceutical composition having immunostimulating activity, comprising a 1,3-diethylbenzimidazolium salt of the general formula according to claim 1 and at least one pharmaceutically acceptable excipient.