RU2160260C1 - 4-oxo-1,4-dihydropyrimide derivative having immunodepressive activity - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: described is new 4-oxo-1,4-dihydropyrimidine, more specifically 2,6-diethyl-5-phenyl -1-(5,6-dimethylbenzylimidazolyl-1)-4-oxo-1,4- di-hydropyrimidine I

Description

 The invention relates to new synthetic derivatives of 4-oxo-1,4-dihydropyrimidine. Derivatives of the 4-oxopyrimidine series are known containing heterocyclic substituents, namely, imidazole, pyrimidine and pyridine together with thiosol [Spengier J. P. Schurack W. // Arch. Pharm-1983-Vol. 316-p. 82-83: C.A.-1983-Vol. 98-125979 f] possessing antihistamine activity.

 Derivatives of 4 (3H) - pyrimidines possessing tranquilizing [Sidney S.G. // Eur. Path.AppI. E.R. 107914 C 07 D 401/02090584; C.A. 1984 -Vol. 101-1919537z] and salts of 4-oxo-1,4-dihydropyrimidine possessing psychostimulant, antidepressant [Cashyap M. M. //J.labelled Compd. Radiophann -1989 -Vol. 22-N 12-p. 1239. 1250; C.A. -1987 -Vol. 106-18475 a], antiallergic and immunostimulating activity [Prediction of biological activity and synthesis of new N-hetero derivatives of 4-oxo-1,4-dihydropyrimidine. Pyatigorsk. 1995, p. 22 - Dep. in VINITI N 2796-B-95 10/23/95].

 The closest in structure to the claimed compound are: 2,6-dimethyl-5-phenyl-1- (5,6-dimethylbenzimidazolyl-1 -) - 4-oxo-1,4-dihydropyrimidine and 2,6-diethyl-5- phenyl-1- (2-methylbenzimidazolyl-1) - 4-oxo-1,4-dihydropyrimidine with immunostimulating activity [Patent N2037491.].

 The aim of this invention is to obtain a new derivative of 4-oxo-1,4-dihydropyrimidine, which has a more pronounced immunosuppressive activity compared to known substances.

 The inventive object is a derivative of pyrimidine 4-oxo-1,4-dihydropyrimidine and its biological properties are not described in the literature available to us. This new substance relates to pyrimidine derivatives containing a heterocyclic substituent and having an immunotropic effect.

обладающий иммунодепрессивной активностью
Описанное соединение получается кипячением N-пропионил-2- фенилацетопропионамида с 1-амино-5,6-диметилбензимидазолом в ледяной уксусной кислоте по схеме:

Пример. Получение 2,6-диэтил-5-фенил-1 -(5,6- диметилбензимидазолил-1)-оксо-1,4-дигидропиримидина. В смеси из 2 мл ледяной уксусной кислоты и 2 мл ДМФА растворяют 2,47 г (0,01 моль) N-пропионил-2-фенилацетопропионамид и 1,61 г (0,01 моль) 1- амино-5,6-диметилбензимидазола. Реакционную смесь кипят 1 час, затем охлаждают и выливают в 100 мл эфира. Выделившийся осадок отфильтровывают и промывают бензолом. Выход 49% (1,99г). Т_пл =149-150^oC (из этанола). Найдено,%: C 74,23; H 6,61; N 15,56; О 3,60. Вычислено %: С 74,21; H 6,53; N 15,64; О 3,62.The goal is achieved as a result of the synthesis of compounds of the following structure:

immunosuppressive
The described compound is obtained by boiling N-propionyl-2-phenylacetropionamide with 1-amino-5,6-dimethylbenzimidazole in glacial acetic acid according to the scheme:

Example. Preparation of 2,6-diethyl-5-phenyl-1 - (5,6-dimethylbenzimidazolyl-1) -oxo-1,4-dihydropyrimidine. 2.47 g (0.01 mol) of N-propionyl-2-phenylacetropionamide and 1.61 g (0.01 mol) of 1-amino-5,6-dimethylbenzimidazole are dissolved in a mixture of 2 ml of glacial acetic acid and 2 ml of DMF . The reaction mixture is boiled for 1 hour, then cooled and poured into 100 ml of ether. The precipitate formed is filtered off and washed with benzene. Yield 49% (1.99g). _Mp = 149-150 ^° C (from ethanol). Found,%: C 74.23; H 6.61; N, 15.56; About 3.60. Calculated%: C 74.21; H 6.53; N, 15.64; About 3.62.

IR spectrum, cm ^-1 : 1705, 1653, 1570, 1535 (liquid paraffin).

UV spectrum λ _max : 207 nm; 252 them; 278 nm (C 1 • 10 ⁵ mol / L in ethanol).

где - P_о - масса опытной лапки;
P_к - масса контрольной лапки,
Введение исследуемого вещества вызывает сдвиг индекса. Исследуемые вещества вводили однократно в дозе 25 мг/кг массы мышей в 1% крахмальной взвеси в желудок с помощью специального зонда за 2 суток до антигенной нагрузки. В качестве объекта сравнения использовали официнальный препарат метилурацил, обладающий иммунотропной активностью. Метилурацил вводили в той же дозе и по той же схеме, что и исследуемое вещество.The study of the immunotropic effect of the 4-oxo-1,4-dihydropyrimidine derivative was carried out on a delayed-type hypersensitivity reaction (HRT) model, which is one of the most commonly used methods for assessing cellular immunity [Immunological methods / Ed. Frimelya G.M. , Medicine, 1987, p. 346 p. 64.]. A total of 18–20 g of CBA lines weighing 18–20 g were immunized intraperitoneally with ram erythrocytes (EB) in an amount of 2 • 10 ⁹ cells / ml. All animals were divided into 3 groups of 25 mice: 1 gr. - control; 2 g - comparison drug - methyluracil; 3 gr. - experience, i.e. research new substance (I). In each group, 5 series of studies were carried out, in which 5 mice were used per series. To determine the HRT reaction, after 4 days, a resolving dose of EB suspension in the amount of 0.01 ml was injected subcutaneously in the right hind paw, and in the other, the control, the same amount of sterile saline. The severity of the HRT reaction to EB was evaluated by the degree of edema of the experimental paw compared to the control by measuring the mass of the paws and the reaction index (IR) was calculated as a percentage

where - P _about - the mass of the experimental paws;
P _to - the mass of the control paw,
The introduction of the test substance causes a shift in the index. The test substances were administered once at a dose of 25 mg / kg of mouse weight in 1% starch suspension in the stomach using a special probe 2 days before antigenic loading. An official preparation methyluracil with immunotropic activity was used as a comparison object. Methyluracil was administered at the same dose and in the same manner as the test substance.

 Control animals received 1% starch suspension in an appropriate amount and were immunized according to the same scheme as the experimental animals.

 The results of the experiments are statistically processed and are presented in table 1.

The effect of substance (I) on the humoral immune response was assessed by the number of antibody-forming cells (AOKs) in the spleen of mice by bezagarovnogo local hemolysis in a monolayer of red blood cells [M.D. Mashkovsky. Medicines - M.: Medicine, 1971, p. 157.]
The experiments were performed on 75 mice of the CBA line of both sexes weighing 20-22 g. All animals were divided into 3 groups of 25 mice: 1 g. - control; 2 g - comparison drug - methyluracil; 3 gr. - experience, i.e. research new substance (I). In each group, 5 series of studies were carried out, in which 5 mice were used per series. Animals were immunized with EB intraperitoneally (2 x 10 ⁹ cells / ml). The number of AOKs in the spleen of mice was determined on the fifth day after immunization and counted on 10 ⁶ splenocytes. 2 days before immunization, the test substance (I) (group 3) and methyluracil (group 2) were injected into the stomach with mice in doses of 25 mg / kg of body weight in 1% starch suspension using a probe. Control animals (group 1) were immunized according to the same scheme as the experimental animals, they were given 1% starch suspension in an appropriate amount. The experimental results are processed statistically and are shown in table 2.

The determination of acute toxicity and the calculation of LD _{50 was} carried out according to the method of Kerber and found that for compound (I) LD ₅₀ is 1500 mg / kg of animal body weight. Thus, the test compound (I) has low toxicity.

 The results of the studies show that the studied compound affects both cellular and humoral immunity.

 Thus, compound (I) inhibits cellular immunity, tested in the HRT reaction to sheep erythrocytes, by 24.6% compared with the control.

 Significant inhibition of humoral immunity was manifested in a decrease in the amount of AOK in the spleen of mice. Compound (I) in its activity depressed antibody production by 30.9% compared with the control.

List of references
1. Spengler JP, Schurack W. // Arch. Pharm. - 1983.- Vol. 316.-p. 82-83; CA - 1983. Vol. 98 - 125979F.

 2. Sidney S.G.//Eur. Path Appl. E. P. 107 914 C 07 D 401/02090584; C.A. 1984 - Vol. 101 - 1919537z.

 3. Cashyap M.M.// J. Labelled. Compd.Radiophann. -1989. - Vol. 22 - N12 p 1239.1250; C.A. - 1987. - Vol. 106 - 18475a.

 4. Forecast of biological activity and synthesis of new N-heteryl derivatives of 4-oxo-1,4-dihydropyridine. Pyatigorsk. - 1995. - S. 22. - Dep. in VINITI N 2796 - B-9523.10.95.

 5. Patent N 2037491.

 6. Immunological methods / Ed. Frimelya G.M., Medicine, 1987, p. 346 p. 64.

 7. M. D. Mashkovsky. Medicines -M.: Medicine, 1971, p. 157.

Claims (1)

Derived 4-oxo-1,4-dihydropyrimidine-2,6-diethyl-5-phenyl-1- (5,6-dimethylbenzimidazolyl-1 -) - 4-oxo-1,4-dihydropyrimidine of the formula I

showing immunosuppressive activity.

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