RU2253445C1 - Immunomodulator - Google Patents

Abstract

FIELD: drugs, medicine.

SUBSTANCE: invention relates to application of 2-hydroxyphenoxyacetic 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, autoimmune and lymphoproliferative disorders due to selective action on cell and humoral immunity.

EFFECT: new drug for treatment of infections, inflammation, and other diseases.

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, pharmacology and clinical immunology have paid great attention to the development of new immunomodulating drugs that stimulate or suppress the humoral and cellular immune responses of the body. Currently existing drugs do not have sufficient selectivity of action on individual parts of the immune system, and also cause the occurrence of undesirable side reactions.

One of the most famous immunomodulators for clinicians is the drug levamisole, which, depending on the dose used, can have both a stimulating and suppressive effect on the immune system, which determines the indications for its use: autoimmune conditions (rheumatoid arthritis, systemic lupus erythematosus, Reiter’s disease and others , requiring suppression of the severity of the body's immune responses), or immunodeficiency, requiring stimulation of the cells of the immune system (Russian Drug Register. - Issue 9. - Article .466-467). However, it should be noted that the use of levamisole can cause various side effects (headache, sleep disturbance, allergic reactions, agranulocytosis, leukopenia, generalized convulsions, damage to the liver and kidneys, etc.), which significantly limits its widespread clinical use.

In order to expand the arsenal of effective and low-toxic synthetic agents with immunomodulating properties, it is proposed to use tris (2-hydroxyethyl) ammonium salt of 2-hydroxyphenoxyacetic acid.

Derivatives of arylheteroalkanecarboxylic acids differ in a variety of biological effects. Tris- (2-hydroxyethyl) ammonium salt of 2-hydroxyphenoxyacetic acid is a stabilizer of cell membranes. (“Clinical efficacy and safety of cephalosporin antibiotics manufactured by LLC“ ABOLmed ”, collected works of NIIKI, ASMU and ICD. Novosibirsk, 2002, p. 179).

The essence of the invention lies in the fact that the effect of tris (2-hydroxyethyl) ammonium salt of 2-hydroxyphenoxyacetic acid on the number of antibody-forming spleen cells in mice (primary and secondary humoral response) and on delayed-type hypersensitivity (cellular immune response) was studied.

We used healthy sexually mature animal mice - hybrids (CBAxC57BL / 6) F1 (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 were processed using the nonparametric U Wilcoxon-Mann-Whitney test.

Example 1. The determination of the number of IgM 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) was administered intraperitoneally to mice once a day daily for 5 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 1 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 2-hydroxyphenoxyacetic acid Groups Dose 1/10 LD ₅₀ (mg / kg) IgM AOK / Splez. Dose 1/100 LD ₅₀ (mg / kg) IgM AOK / Splez. Control (n = 10) 1839 1839 Compound (n = 10) 300 19881 * thirty 773 * * significantly, P <0.05; n is the number of animals.

As can be seen from table 1, tris- (2-hydroxyethyl) ammonium salt of 2-hydroxyphenoxyacetic acid significantly stimulates the primary humoral response in a dose of 1/10 LD ₅₀ and suppresses in a dose of 1/100 LD ₅₀

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

The compound at a dose of 1/10 LD ₅₀ and 1/100 LD ₅₀ in a volume of 0.5 ml was administered intraperitoneally once a day daily for 5 days. The control animals in the same volume and mode were injected with a solvent of the compounds (water). On the day of the last administration of the compounds, primary immunization with 0.25% EB in a volume of 0.5 ml was performed intraperitoneally. 30 days after the primary immunization, secondary EB immunization was performed at a dose of 0.5 × 10 ⁷ / mouse intravenously. The amount of AAG IgG was determined in the spleen at the peak of the immune response (on the 5th day after secondary immunization) by local hemolysis (Cunningham, 1968: Dresser, Wortis, 1965). Spleen cells were incubated for 2 hours at 39 ° C in chambers with EB, complement of guinea pig and rabbit antiserum against mouse IgG (2000-fold dilution). Hemolysis zones were counted under x42 magnification. The results were expressed as the absolute amount of AOG IgG in the spleen.

table 2 Absolute amount of AOK IgG in the spleen of intact CBF1 mice under the influence of the course administration of tris (2-hydroxyethyl) ammonium salt of 2-hydroxyphenoxyacetic acid at a dose of 1/10 and 1 / 100LD ₅₀ . Groups Dose 1/10 LD ₅₀ (mg / kg) IgG AOK / Sles. Dose 1/100 LD ₅₀ (mg / kg) IgG AOK / spleen. Control (n = 12) 71928 71928 Compound (n = 12) 300 84027 thirty 47188 * * significantly, P <0.05; n is the number of animals.

As can be seen from table 2, the compound at a dose of 1 / 100LD ₅₀ significantly inhibits the formation of IgG AOK, while at a dose of 1/10 LD _{50 it} does not affect the number of Ig AOK (there is a tendency to stimulation).

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 percentages or absolute values.

Table 3 The effect of tris- (2-hydroxyethyl) ammonium salt of 2-hydroxyphenoxyacetic acid on delayed-type hypersensitivity. Groups HRT (%) 1. Control (n = 10) 47.7 2. Compound 30 mg / kg (n = 10) 42.5 3. The compound at a dose of 300 mg / kg (n = 10) 30.4 * * significantly, P <0.05; n is the number of animals.

As can be seen from table 3, the compound significantly inhibits the cellular immune response (HRT) only at a dose of 1/10 LD ₅₀ .

Evaluating the obtained results in general, it can be noted that the test compound significantly inhibited a 1 / 100LD ₅₀ IgM and IgG response, without affecting HRT, and at a dose of 1/10 LD _{50, it} significantly stimulated the IgM response, but suppressed HRT, t .e. possesses sufficient selectivity of action on the humoral and cellular immune response, depending on the dose used. The presence of these properties in the tris (2-hydroxyethyl) ammonium salt of 2-hydroxyphenoxyacetic acid will make it possible in the future to create drugs based on it for the treatment of various immunocompromised conditions.

Claims (1)

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