RU2294199C2 - Maleic acid 3-methyl-2-pyridylamide eliciting direct anticoagulant effect - Google Patents

Abstract

FIELD: organic chemistry, chemical technology, medicine, hematology.

SUBSTANCE: invention relates to using maleic acid 3-methyl-2-pyridylamide of the formula:

in medicine as an anticoagulant agent. Compound is synthesized by interaction of 2-amino-3-methylpyridine with maleic anhydride. Synthesized compound is a white crystalline substance with value T_m = 136-137°C and soluble in water, ethanol, dimethylformamide (DMFA) and dimethylsulfoxide (DMSO). Synthesized substance provides increasing coagulating time of citrate blood and percent for coagulating capacity change in the concentration of this substance 1 mg/ml and 0.5 mg/ml by 502.4% and 92.2%, respectively. Value LD₅₀ for this substance is 20.5 (12.2-31.3) mg/kg after its intravenous injection in white mice.

EFFECT: valuable medicinal property of compound.

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Description

The invention relates to the use of biologically active compounds, namely, maleic acid 3-methyl-2-pyridylamide (I) of the formula

as a means of having a direct anticoagulant effect, which suggests its use in medicine as a direct anticoagulant.

Known maleic acid 2-pyridylamide of the formula

obtained and described by us earlier [Kolotova N.V., Kozminykh V.O., Dolzhenko A.V., Kozminykh E.N., Kotegov V.P., Godina A.T., Syropyatov B.Ya., Novoselova G .N., Chem.-farm. Journal, T.35, No. 3, p.26-30 (2001); Dolzhenko A.V., Kolotova N.V., Kozminykh V.O., Syropyatov B.Ya., Chem. Journal, T.36, No. 3, S.17-19 (2001)].

Compound II exhibits a weak hypertensive effect; data on its anticoagulant activity are not available in the literature.

Only one direct-acting anticoagulant is known and used in medical practice - heparin [Mashkovsky M.D. Medicines (a manual for doctors), M .: "Publishing House of Medicine", T.2, C.73, (1985)], which we have taken as a standard for comparing the anticoagulant activity of compound I.

The aim of the invention is to search for compounds with direct anticoagulant activity among known maleic acid heterylamides. This goal is achieved by the use of 3-methyl-2-pyridylamide maleic (I) acid [Dolzhenko A.V., Syropyatov B.Ya., Kozminykh V.O., Kolotova N.V., Zakhmatov A.V., Borodin A. YU. Chem.-farm. Journal, T.37, No. 8, C.12-13 (2003)], which is obtained by acylation of 2-amino-3-methylpyridine with maleic anhydride.

The method of obtaining 3-methyl-2-pyridylamide maleic acid. To a solution of 1.08 g (0.01 mol) of 2-amino-3-methylpyridine in 30 ml of dioxane, 0.98 g (0.01 mol) of maleic anhydride in 40 ml of dioxane was added with stirring at room temperature. After 1 h, the precipitate formed is filtered off, dried and crystallized from ethanol. Yield 1.65 g (80%). T _pl. = 136-137 ° C.

Compound (I) is a white crystalline substance, soluble in water, ethanol, DMSO, DMF. The ¹ H NMR spectrum ("РЯ-2310" (60 MHz) in DMSO-d ₆ , internal standard - HMDS) of compound I contains: a singlet of three protons of the methyl group at 2.18 ppm, a singlet of two protons of the group СН = CH at 6.22 ppm, the multiplet of three protons of the pyridine ring system and the proton signal of the amide group CONH in the range of 6.58-8.05 ppm and a broadened signal of one proton of the carboxyl group in the region of 13.58 ppm

Compound I was tested for direct anticoagulant activity. Experiments on the study of anticoagulant action were carried out using a Minilab 701 coagulometer. For the study, citrate (3.8%) blood (9: 1) of the dog was used. The effect of compound I was studied at concentrations of 1 mg / ml, 0.5 mg / ml and 0.25 mg / ml blood. Heparin was tested at a concentration of 1 U / ml of blood. In each series of experiments, eight animals were used. The degree of anticoagulant activity of the test compound was determined by the change in the coagulation time of citrate blood and the percentage change in coagulation in the control and experiment and was statistically processed using the Student coefficient [Sernov L.N., Gatsura V.V. Elements of experimental pharmacology, M., S.312-313, (2000)]. The test results are presented in the table.

As can be seen from the table, the test compound exhibits a dose-dependent direct anticoagulant effect, because significantly increases the coagulation time of citrate blood depending on its concentration at concentrations of 1 mg / ml and 0.5 mg / ml and reduces coagulation, respectively, by 502.4% and 92.2%. The reference standard heparin at a concentration of 1 IU / ml of blood inhibits coagulation by 22.4%.

Acute toxicity after intravenous administration was determined on nonlinear white mice weighing 20-25 g. The test substance was injected into the tail vein in the form of an aqueous solution at the rate of 0.1 ml per 10 g of animal weight in increasing doses. The results were processed according to Prozorovsky with the calculation of the average lethal dose (LD ₅₀ ) at P = 0.05 [Prozorovsky VV, Prozorovskaya MP, Demchenko VM Farmakol. Toxicol., T.41, No. 4, C.497-502 (1978)].

The acute toxicity of the claimed compounds was 20.5 (12.2-31.3) mg / kg

Thus, maleic acid (I) 3-methyl-2-pyridylamide exhibits a dose-dependent direct anticoagulant effect and can be used in medicine as a direct anticoagulant.

Literature

1. Dolzhenko A.V., Kolotova N.V., Kozminykh V.O., Syropyatov B.Ya., Chem. Journal, T.36, No. 3, S.17-19 (2001).

2. Kolotova N.V., Kozminykh V.O., Dolzhenko A.V., Kozminykh E.N., Kotegov V.P., Godina A.T., Syropyatov B.Ya., Novoselova G.N., Chem.-farm. Journal, T.35, No. 3, C.26-30 (2001).

3. Mashkovsky M.D. Medicines (a manual for doctors), M .: "Publishing House of Medicine", T.2, S.73 (1985).

4. Dolzhenko A.V., Syropyatov B.Ya., Kozminykh V.O., Kolotova N.V., Zakhmatov A.V., Borodin A.Yu. Chem.-farm. Journal, T.37, No. 8, S.12-13 (2003).

5. Sernov L.N., Gatsura V.V. Elements of experimental pharmacology, M., S.312-313, (2000).

6. Prozorovsky V.V., Prozorovskaya M.P., Demchenko V.M. Farmakol. Toxicol., T.41, No. 4, C.497-502 (1978).

Table The effect on the blood coagulation system of various concentrations of the claimed compound (I) and the reference standard (heparin) Compound concentration Coagulation time, sec. The control Coagulation time, sec.
Experience % coagulation changes R Compound I 41.4 ± 3.90 247.6 ± 39.62 -502.4 <0.001 1 mg / ml (n = 8) Compound I 32.2 ± 4.15 61.9 ± 6.89 -92.2 <0.001 0.5 mg / ml (n = 8) Compound I 22.2 ± 2.29 26.8 ± 1.95 -20.7 > 0.05 0.25 mg / ml (n = 8) Heparin 29.9 ± 0.48 36.6 ± 1.82 -22.4 <0.01 1 U / ml (n = 8) Note: P - compared to control

Claims (1)

The use of 3-methyl-2-pyridylamide maleic acid as a means of having direct anticoagulant action.