RU2253447C1 - Immunomodulator - Google Patents

Abstract

FIELD: drugs, medicine.

SUBSTANCE: invention relates to application of 4-chlorophenoxyacetic acid tris-(2-hydroxyethyl)ammonia salt, which represents known antioxidant, as immunomodulator. Present invention makes it possible to produce drugs for treatment of infections, inflammation, and autoimmunity disorders, wherein stimulating action to leukocyte amount and primary humoral response is combined with hypersensitivity inhibiting.

EFFECT: new drug for treatment of infections, inflammation, and autoimmune disorders.

3 tbl, 3 ex

Description

The invention relates to new biologically active substances from the class of alkane carboxylic acids and can be used in medicine and biology as the basis for the creation of drugs

In recent years, immunopharmacology pays much attention to the development and study of specific agents that stimulate or suppress the body's immune responses. Of particular interest in this regard are chemical compounds that can affect immunity in different directions, for example, stimulate humoral and inhibit cellular reactions, i.e. manifest themselves simultaneously as an immunostimulant and immunosuppressant.

The well-known immunomodulator of synthetic nature Levamisole (M.D. Mashkovsky. Medicines. - M., 2001. P.194-195) can perform the functions of an immunomodulator that can enhance a weak reaction of cellular immunity, weaken a strong one and not act on a normal one. Levamisole has a dose-dependent effect on the immune system, and it is necessary to use it, for example, as an immunostimulant with caution, since when doses are exceeded, it is possible not to have an immunostimulating, but a pronounced immunosuppressive effect against all immunity reactions, and in some cases, from small doses of levamisole. In addition, the use of levamisole causes various side effects (fever, headache, sleep disturbance, allergic reactions, agranulocytosis, leukopenia, etc.).

Modern knowledge about the pathogenesis of many diseases associated with the disruption of interclonal relationships of immunocompetent cells and the balance of cytokines produced by them put forward urgent requirements for the search for new immunoactive drugs. In this regard, drugs are of interest that exhibit a multidirectional effect on integral indicators, such as antibody formation and HRT, during primary in vivo screening for intact animals, affecting either humoral or cellular immunity only and not toxic to the body.

In order to expand the arsenal of synthetic agents with immunomodulating properties, it is proposed to use the tris (2-hydroxyethyl) ammonium salt of 4-chlorophenoxyacetic acid as an immunomodulator.

Tris- (2-hydroxyethyl) ammonium salt of 4-chlorophenoxyacetic acid and its analogues have antioxidant properties (“Clinical efficacy and safety of cephalosporin antibiotics manufactured by ABOLmed LLC. Proceedings of NIIKI, ASMU and ICD. - Novosibirsk, 2002. P.179. ; www.abolmed.ru/pages/article33.htm).

The essence of the invention lies in the fact that the immunomodulating properties of tris (2-hydroxyethyl) ammonium salt of 4-chlorophenoxyacetic acid were studied by the ability of this compound to affect the number of nucleated blood cells in mice (myeloactive properties), the number of antibody-forming cells (IgM AOK) in vivo (primary humoral immune response) and delayed-type hypersensitivity (cellular immune response).

We used healthy sexually mature animal mice - hybrids (CBAxC57BL / 6) Fl (CBF1) of both sexes, 8–10 weeks old, weighing 18–20 g. The spread in the groups according to the initial body weight did not exceed ± 10%. Control and experimental animals of the same age and obtained simultaneously from the same cattery ("Dawn", Tomsk). Before and during the experiment, control and experimental animals were kept in a vivarium under the same conditions: standard plastic cages with small wood shavings (no more than 10 individuals) on a standard diet. All studies were carried out at the same time of the day (in the morning). The data obtained were processed according to the nonparametric criterion U of Wilcoxon-Mann-Whitney.

Example 1. Determination of myeloactive properties.

In an experimental group of animals, the compound was administered at a dose of 1/10 LD 50 intraperitoneally in a volume of 0.5 ml of solvent daily for 7 days. 24 hours after the last administration, the number of nucleated cells in the peripheral blood was determined. The control animals in the same volume and mode were injected with a solvent (water). The criterion for assessing myeloactive properties is a change in the leukocyte formula of ± 20%. The results were expressed as the number of leukocytes in one ml of blood and as a percentage of the corresponding control.

Table 1 Effect of Tris (2-hydroxyethyl) ammonium salt of 4-chlorophenoxyacetic acid on the number of nucleated blood cells in intact CBF1 mice. Group Dose 1/10 LD ₅₀ (mg / kg) Number of nucleated cells % to control Control (n = 14) 9.87 × 10 ⁶ / ml Compound (n = 15) 150 12.2 × 10 ⁶ / ml * +24 * significantly, P <0.05; n is the number of animals.

As can be seen from table 1, the compound reliably has myelostimulating properties (causes an increase in the number of leukocytes by 24%).

Example 2. The determination of the number of Ig M antibody-forming cells in vivo.

The compound at a dose of 1/10 LD ₅₀ and at a dose of 1/100 LD ₅₀ in a volume of 0.5 ml (solvent) was administered intraperitoneally to experimental animals once a day daily for 7 days. The control animals in the same volume and mode were injected with the solvent of the compound (water). On the day of the last administration, the compounds of animals were immunized intravenously with ram erythrocytes (EB) at a dose of 0.5 × 10 ⁷ / mouse. The amount of IgM AOK in the spleen of mice was evaluated on the 4th day after immunization by the number of local hemolysis zones in a semi-liquid medium using the modified Cunningham method (1968). The results were expressed as the absolute amount of IgM AOK in the spleen.

table 2 The absolute amount of IgM AOK in the spleen of intact CBF1 mice under the influence of the course administration of tris- (2-hydroxyethyl) ammonium salt of 4-chlorophenoxyacetic acid Groups Dose 1/10 LD ₅₀ (mg / kg) IgM AOK / spleen. Dose 1/100 LD ₅₀ (mg / kg) IgM AOK / spleen. Control (n = 15) 1839 1839 Compound (n = 15) 150 11138 * fifteen 1481 * significantly, P <0.05; n is the number of animals

As can be seen from the data in Table 2, the tris (2-hydroxyethyl) ammonium salt of 4-chlorophenoxyacetic acid exhibits a pronounced stimulating effect on the primary humoral response at a dose of 150 mg / kg.

Example 3. The reaction of delayed-type hypersensitivity (HRT) in vivo.

In an experimental group of animals, the compound was administered at a dose of 1/10 LD ₅₀ intraperitoneally daily for 5 days in a volume of 0.5 ml. On the day of the last administration of the compound, the mice were sensitized by intraperitoneal administration of 0.25% EB in a volume of 0.5 ml; on the 4th day after sensitization, a resolving dose of antigen was administered under the plantar aponeurosis of the right hind paw (50% EB in a volume of 50 μl). The solvent was injected into the contralateral paw in the same volume. The control animals in the same volume and mode were injected with the solvent of the compound (water). HRT response was evaluated by local HRT (Crowie, 1975). The reaction was taken into account 24 hours after the introduction of the resolving dose of EB, the magnitude of the edema was evaluated with a caliper. The results were expressed as a percentage.

Table 3 Effect of Tris (2-hydroxyethyl) ammonium salt of 4-chlorophenoxyacetic acid on delayed-type hypersensitivity Groups HRT (%) 1. Control (n = 14) 47.5 2. Compound 150 mcg / kg (n = 15) 23.4 * * significantly, P <0.05; n is the number of animals

As can be seen from table 3, the compound at a dose of 150 mg / kg significantly inhibits the cellular immune response.

Assessing the overall results, it can be noted that, according to the effect on the myeloid activity and the primary humoral response, tris- (2-hydroxyethyl) ammonium salt of 4-chlorophenoxyacetic acid can be classified as immunostimulants, and by its effect on the cellular immune response, to immunosuppressants.

The examples given illustrate the presence of tris- (2-hydroxyethyl) ammonium salt of 4-chlorophenoxyacetic acid with immunomodulatory properties, which will make it possible in the future to develop drugs based on it for treating various immunocompromised conditions.

Claims (1)

The use of tris- (2-hydroxyethyl) ammonium salt of 4-chlorophenoxyacetic acid as an immunomodulator.