RU2312660C2 - 4-acetyl-3-hydroxy-1-(2-hydroxyethylaminoethyl)-5-phenyl-3-pyrrolin-2-one eliciting anti-coagulant activity - Google Patents

Abstract

FIELD: organic chemistry, hematology.

SUBSTANCE: invention proposes an agent decreasing blood coagulation representing 4-acetyl-3-hydroxy-1-(2-hydroxyethylaminoethyl)-5-phenyl-3-pyrrolin-2-one of the formula (1):

Agent elicits direct anti-coagulant effect as it reliably increases the time coagulation value of citrate blood by 30.4%.

EFFECT: valuable medicinal property of agent.

1 tbl, 1 ex

Description

The invention relates to a new biologically active substance of the series 3-hydroxy-3-pyrrolin-2-one, namely to 4-acetyl-3-hydroxy-1- (2-hydroxyethylaminoethyl) -5-phenyl-3-pyrrolin-2-one formulas:

possessing anticoagulant activity, which suggests the possibility of using it in medicine as a drug that effectively reduces blood coagulation.

The closest structural analogue of the claimed compound is 3-hydroxy-1- (3-carboxypropyl) -4-methylsulfonyl-5-phenyl-3-pyrrolin-2-one (Kataeva A.V. Synthesis, properties and biological activity of 1-substituted 5- aryl-3-hydroxy-4-methylsulfonyl-3-pyrrolin-2-ones: diss .... Candidate of Pharmaceutical Sciences / Perm State Pharmacist Acad. - Perm, 2000. - P.60) formulas:

which is taken as a prototype and does not have anticoagulant effect. The comparison standard is heparin (Mashkovsky MD Medicines: practical aid. - 15th ed. - M .: "New Wave", 2005. - P.475), which is used in medical practice and is an analogue in action.

The essence of the invention is the search in a series of 3-hydroxy-3-pyrrolin-2-ones for compounds with pronounced anticoagulant activity that reduces blood coagulation. This is achieved by the synthesis of 4-acetyl-3-hydroxy-1- (2-hydroxyethylaminoethyl) -5-phenyl-3-pyrrolin-2-one, which has an anticoagulant effect.

The inventive compound is obtained by the interaction of methyl ester of acetylpyruvic acid, benzaldehyde and 2-hydroxyethylaminoethylamine according to the scheme:

An example of obtaining the claimed compounds.

Example 1

4-acetyl-3-hydroxy-1- (2-hydroxyethylaminoethyl) -5-phenyl-3-pyrrolin-2-one

To a solution of 1.02 ml (0.01 M) of benzaldehyde and 1.01 ml (0.01 M) of 2-hydroxyethylaminoethylamine in 5 ml of 1,4-dioxane at room temperature, a solution of 1.44 g (0.01 M) of acetylpyruvic acid methyl ester in 5 ml of 1,4-dioxane is added and left at the same temperature for 1 day. The precipitate formed is filtered off, recrystallized from ethanol and dried. Yield 1.76 g (58%). Mp: 180-182 ° C. Found,%: C 63.08; H 6.47; N 9.15. C ₁₆ H ₂₀ N ₂ O ₄ . Calculated,%: C 63.14; H, 6.62; N, 9.21.

The inventive compound is a colorless crystalline substance, soluble in water, ethanol, dimethylformamide, dimethyl sulfoxide.

IR spectrum (SPECORD, liquid paraffin, ν, cm ^-1 ): 1648 (СО), 1706 (CON), 3150 (С ₍₃₎ OH), 3360 (NH), 3420 (AlkOH).

), 3.03 (2Н, m, C₍₄₎H₂), 3.12 (1H, m,

, 3.25 (1Н, m,

), 3.68 (1Н, m,

), 3.73 (2Н, m, C₍₅₎H₂), 5.13 (1Н, s, C₍₅₎H), 7.16 (3Н, Н_аром), 7.24 (2Н, Н_аром), 9.40 (1Н, b.s, NH).1 H-NMR spectrum (Bruker DRX500, 500.13 MHz, DMSO-D ₆ , TMS, δ, ppm): 2.11 (3H, s, CH ₃ CO), 2.74 (1H, m,

), 3.03 (2H, m, C ₍₄₎ H ₂ ), 3.12 (1H, m,

3.25 (1H, m,

), 3.68 (1H, m,

), 3.73 (2H, m, C ₍₅₎ H ₂ ), 5.13 (1H, s, C ₍₅₎ H), 7.16 (3H, H _arom ), 7.24 (2H, H _arom ), 9.40 (1H, bs , NH).

The inventive compound and analogue in structure were examined for the presence of anticoagulant activity. The experiments 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 the compounds on blood coagulation was tested at a concentration of 1 mg / ml of blood. A heparin solution was used as a reference for anticoagulant activity, which 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 compounds was determined by the change in the coagulation time of nitrate blood in comparison with the control and the standard. The results were processed using Student's coefficient (Sernov L.N., Gatsura V.V. Elements of experimental pharmacology. M., 2000, pp. 313-313). The test results are presented in the table.

As can be seen from the table, the claimed compound exhibits a direct anticoagulant effect, since it significantly increases the coagulation time of citrate blood by 30.4%. Its structural analogue does not have a reliable anticoagulant effect. Benchmark heparin slows down coagulation by 22.4%.

The acute toxicity of the claimed compound was determined on outbred white mice of both sexes weighing 16-18 g. The compound was administered intraperitoneally and intravenously 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 DL ₅₀ at P = 0.05 (Prozorovsky V.B. Express method for determining the average effective dose and its errors. / VB Prozorovsky, MP Prozorovskaya, V.M. .Phemchenko. // Pharmacol. And Toxicol. - 1978. - T.41, No. 4. - S.497-502).

It was found that acute toxicity (DL ₅₀ ) of the claimed compound with intraperitoneal administration is 1460.0 (610.4-2734.4) mg / kg, and with intravenous administration - 467.2 (195.3-875.0) mg / kg .

Table The effect on the blood coagulation system of the claimed compound and the reference standard Compound Coagulation time, s % coagulation changes R the control experience 4-acetyl-3-hydroxy-1- (2-hydroxyethylaminoethyl) -5-phenyl-3-pyrrolin-2-one 35.5 ± 1.44 46.3 ± 2.75 -30.4 <0.01 3-Hydroxy-1- (3-carboxypropyl) -4-methylsulfonyl-5-phenyl-3-pyrrolin-2-one 40.2 ± 2.63 46.7 ± 2.69 -16.2 > 0.05 Heparin 29.9 ± 0.48 36.6 ± 1.82 -22.4 <0.01

This work was supported by the Russian Foundation for Basic Research (project No. 04-03-96042).

LITERATURE

1. Kataeva A.V. Synthesis, properties and biological activity of 1-substituted 5-aryl-3-hydroxy-4-methylsulfonyl-3-pyrrolin-2-ones: diss .... cand. pharmacist. sciences. / Perm. state pharmacist. Acad. - Perm, 2000 .-- S.60.

2. Mashkovsky M.D. Medicines: pract. allowance. - 15th ed. - M.: "New Wave", 2005. - S. 475.

3. Sernov L.N., Gatsura V.V. Elements of experimental pharmacology. M., 2000, p. 312-313.

4. Prozorovsky VB Express method for determining the average effective dose and its errors. / V.B. Prozorovsky, M.P. Prozorovskaya, V.M.Demchenko. // Farmakol. and toxicol. - 1978. - T.41, No. 4. - S.497-502.

Claims (1)

The use of 4-Acetyl-3-hydroxy-1- (2-hydroxyethylaminoethyl) -5-phenyl-3-pyrrolin-2-one as an agent with anticoagulant activity.