RU1389235C - Derivatives of thieno-[2,3-d]-pyrimidine showing anti- herpetic and antituberculosis activity - Google Patents

Abstract

SUBSTANCE: invention relates to substituted pyrimidine, in particular, thieno I2,3-d pyrimidino8 of the general formula CH-O-C N-CH N-C C-S-CR CR where e 1I1 2 a) R -CH, and R-NO. b) R-NO, and R-CH, oba13221223 giving antipyretic and anti-tuberculous activity), which can be used in medical practice. The purpose of the invention is the creation of new more active substances with an extended spectrum of action. Their synthesis is carried out from the corresponding 3,4-dihydrothieno 2,3-d | pyrimide - 4 - she and ROS in the presence of dimethylformamide followed by boiling in methanol. Yield%: t, gross ph; a) 50, 119 - 120; b) 55; 95 -96. C H N O S. The new substances are active against f and 11 types of herpes at a concentration of 1 μg / ml against 5 μg / ml for the known (chemotherapeutic index 5); the minimum inhibitory concentration for tuberculosis is 3 and 0.55 µg / ml versus 1-2 MU- / ML for areptomycin. 3 tapp

Description

FIELD OF THE INVENTION This invention relates to novel derivatives of THeHo 2,3-d nHpHMHAMHa having biological activity, which can be used in medicine.

The purpose of the invention is the creation of new derivatives of thieno (2,3-th) pyrimidine with higher antiherpetic activity in combination with anti-tuberculosis activity.

Example 1. Preparation of 4-methoxy-5-methyl-6-nitrothieno {2,3-d pyrimidium a (1 a).

Preparation of the starting material, 4-chlorop-5-methyl-6-nitrothieno 2.3-d} pyrimidine (2a).

A mixture of 1.7 g {0.008 mol) of 5-methyl-6-nitpo-3.4-dihydrothienoi 2.3 d pyrimid-4-one, 1 ml (0.01 mol) of phosphorus oxychloride, 0.84 ml (0.01 mol ) dimethylformamide (DMF) in 30 ml dry dichloroethane (DCE) kipT1BT with stirring for 30 minutes. The reaction solution was washed with water, DCE was evaporated in vacuo, the residue was crystallized from a mixture of hexane and methylene chloride (1: 1).

1.5 g (82%) of 4-chloro-5-methyl-b-nvltrothieno 2,3-d pyrimidine are obtained in the form of a yellow crystalline powder; so pl. 134-135 ° C. Found,%; C 36.5; H 1.9; C1 15.2; N, 18.2; S 14.1.

C7H4CIN302S

Calculated,%; C 36.6; H 1.8; C1 15.4; N, 18.3; S 14.0.

Nuclear Magnetic Resonance Spectrum (СНС1з, int.st. TMS). 6, M.D .; 9.15 (s, 2H). 2.52 (s, 5СНе).

Obtaining the target product - 4-methoxy-5-methyl-6-nitrotitno {2,3-d pyrimidine (1a).

A solution of 2.29 g (0.01 mol) of compound 2a in 40 ml of methanol was boiled for 30 minutes. The solution is cooled, the precipitate is filtered off,

Get 1.12. g (50%) of 4-methoxy-5-methyl-6-nitrothieno 2,3-d pyrimidine in the form of a crystalline yellow powder, soluble in chlorohydrocarbons, alcohols, mp, 119-120 ° C,

Found,%: C42.4; H 3.0; N, 18.7; S 14.1.

SHIPPING

Calculated. %; C 42.6; H 3.1; N, 18.6; S 14.2.

Example 2. Preparation of 4-methoxy-5-nitro-6-methylthieno 2,3-d pyrimidine (16).

The preparation of the starting material, 4-chlorop-5-nitro-6-methylthiene 2,3-d pyrimidine (26), was carried out under the conditions of Example 1.

From 3.8 g (0.018 mol) of 5-nitro-6-methylthi-, 3-d nirimide-4-one. 1.65 ml of POCl3, 1.39 ml of DFMA in 100 ml of dry DCE give 2 g (70%) of 4-chloro-5-nitro-6-methyltia- .3-d pyrimidine in powder form

cream color, soluble in alcohols, chlorohydrocarbons, dioxane, ketones. so pl. 84-85 ° C (methylene chloride; hexane 1; 1).

Found; %; C 36.6; H 1.8; C 15.6; N, 18.4; 514.1,

C7H4CIN302S

Calculated,%; C 36.6; H 1.8; C1 15.4; N t8.3; S 14.0.

Nuclear Magnetic Resonance Spectrum (СНС1з, int.st. TMS), b. m, d .; 8.93 (s, 2H); 275 (s. 6-CH3).

Obtaining the target product is 4-methoxy-5-nitro-6-methylthieno {2, 3-d pyrimidine

(15) - is carried out analogously to example 1 of

1.12 g (0.005 mol) of compound 2a in 30 ml

methanol.

0.65 g (55%) of 4-methoxy-5-nit-pp-6-methylthieno 2,3-d pyrimidine is obtained in the form of a white powder with a cream tint, so pl. 95-96 ° C.

Found,%; C42.7; H 3.2; N, 18.9; S 14.3. SAN UMZOZZ

Calculated,%; C 42.6; H 3.1; N, 18.6; S 14.2.

The subject compounds have been studied in cell culture experiments to suppress the multiplication of antigenic herpes simplex viruses type I and II.

To study the antiherpetic activity of the compounds, a primary trilsinized chicken embryo fibroblast cell culture (FEC) was used, which was obtained by trypsinization of 9-day-old chicken embryos. Herpes simplex viruses I and II of antigenic types (strains L2 and TP) were used as test viruses.

A two-day monolayer of cell culture was infected with virus dilutions containing 10 to 1000 TCDbo in 0.4 ml. After adsorption and removal of the non-adsorbed virus (1 h after the inoculation of the virus), the studied compounds were introduced in concentrations of 1/2 of the maximum tolerated for cell culture

concentrations, and in lower concentrations.

The results were taken into account after 48 h according to the ability of the studied compounds to prevent the cytopathic effect of the virus on cells and expressed by the amount of 50% inhibited tissue cytopathic doses of the virus (TCDbo).

The study of the virus-inhibiting activity of the compounds of the invention in relation to herpes simplex viruses was carried out on

1,600 tubes with a monolayer of FEC cell culture; statistical processing results.

The anti-tuberculosis activity of the compounds of the invention was studied in vitro, determining the minimum inhibitory concentration (MIC) by serial dilution using Soton medium. As a test culture used mycobacterium tuberculosis of the human type (M. tuberculosis H37Rv). The cultivation time of the tested concentrations of compounds with test cultures was 14 days at 37 ° C.

The toxicity of the compounds of the invention was studied in experiments on cell culture FEC (96 tubes) and white non-linear mice weighing 18-20 g (120 mice). Nutrient medium No. 199 containing various concentrations of the test compound was applied to a monolayer of a 48-hour cell culture of FEK and microscopy of cell cultures was carried out for 5 days to reveal the cytotoxic effect of the compounds and determine the concentration maximum tolerated by the cells.

In mice, test compounds were administered orally as a suspension in a 1% starch gel in a volume of 0.5 ml once daily for 5 days. The survival of the animals was monitored for 14 days and the maximum tolerated dose (MTD) of the compound was determined.

 The results of a study of the antiherpetic activity of the compounds are given in table. 1 and 2.

The comparison was carried out with a known compound of formula A

N

possessing a virus-inhibiting effect against the herpes simplex virus in the cell culture of FEC,

The results of a study of the compounds of the invention indicate that these compounds have pronounced virigin inhibitory activity against the herpes simplex virus. Compound 1a at concentrations of 2.5 and 2.0 μg / ml feed5

0 5 0

5

0

5

0

5

it reproduces the herpes simplex virus type I on 1.0-2.0 Ig TCDyo. and type II 1.0-1.5 Ig TCDc). Compound 16 is effective only against type I herpes virus: at a concentration of 10 and 5 µg / ml, its reproduction is suppressed by 2.0-1.0 Ig TCDM as compared to the control. Compound A is inferior in antiherpetic activity to the compounds of the invention with respect to herpes simplex virus type I and inactive against herpes virus type II.

The results of a study of the anti-tuberculosis activity of the compounds are given in Table. 3.

As follows from the table. 3, the compounds of the invention, in addition to antiviral activity, have high anti-tuberculosis activity against human tuberculosis mycobacteria, similar in activity to the known anti-tuberculosis antibiotic streptomycin.

The toxicity studies of the compounds of the invention indicate that they are well tolerated by animals: when administered per os once a day for 5 days, the MTD for mice of compounds 1a and 1b is 500 mg / kg, which is higher than the MTD of compound A in 4 times and indicates less toxicity of the compounds of the invention.

An analysis of the results shows that the compounds of the invention have a more pronounced effect and less toxicity compared to the compound of formula A. In addition, the compounds of the invention have a high anti-tuberculosis activity comparable to that of the known antibiotic streptomycin.

(66) Kharizomenova I.A. et al. Synthesis and antiviral activity of derivatives of 3-aminothieio 2.3-b pyrimidine, Chem. Pharmaceutical Journal. 1981, No. 9, p. 40,

Mashkovsky M, D. Medicines M .: Medicine, 1984, v. 2.p.230.

Table 1

 CTI is the reciprocal of the ratio of the minimum active concentration to the maximum tolerated p 0.05,, 01.

Compound

1 a

1 b

Streptomycin

table 2

Table 3

inhibitory concentration, μg / ml

3: 5

0.55 1.0-2.0