SU1124559A1 - 1-(2-oxyethoxymethyl)-1,2,4-triazole-5-carboxamide possessing activity against herpes virus - Google Patents

Description

The invention relates to the field of organic chemistry, medical chemistry and virology, specifically, the eurbatch relates to a new compound I-C2-hydroxyethoxymethyl) -1,2,4-triazole 5-carboxamide of the formula I G jLcr 0 -NH2 CH20CH2Sh20N which is a new antiviral substance selectively repressing the herpes simplex virus. The invention can be used in virology to suppress the reproduction of the herpes virus. Known as the antiviral drug ribavirin (virazole, 1, 1 -) -ribufuranosyl, 2,4-triazole-3-carbox mid, which is a structural analogue of guanosine, Ribavirin has a moderate ability to suppress the reproduction of herpes virus. In general, the number of researches is N-J- (1QQC2 1. MaN 2. СНзСООСН2СН Т CJ-COOC2H5 3 / methanol СН20СН2 СН20ССНз

The alkylation reaction of the ethyl ester of I, 2,4-triazole-3-carboxylic acid obtained according to the well-known scheme gives 1- (1-acetoxyethoxymethyl-1,2, i -triazole-5-carboxylic acid ethyl ester, treatment of which with a semi-dilute ammonia solution in methanol at room temperature leads to the production of the desired product, isolated by known methods.

Example 1. Synthesis of 1-C2-hydroxy. Ethoxymethyl / -, 2,4-triazole-5-carboxamide and its isolation.

To a solution of 5.64 g (40 mmol) of 1,2,4-triazole-ZcarbLnoyl ethyl ester in 160 ml of absolunthooro dimethylformamide is added

1.16 g (48 mmol of sodium hydride, stirred for 30 minutes at room temperature until the hydrogen bubbles disappear) and add

9.16 g (60 mmol) -acetoxyethoxymethyl chloride. The reaction mixture is stirred for 24 hours at room temperature, evaporated on a rotary evaporator, 100 ml are added to the residue.

water and extract the aqueous solution with chloroform. The chloroform solution is dried over, the solution is filtered and evaporated. To the residue obtained is added 200 ml of half-diluted

saturated at O With a solution of dry ammonia in absolute methanol, incubated 48 hours at room temperature and evaporated. The isolation and 592 scientific research institutes made repeated attempts to enhance its antiviral properties through various chemical modifications, which, however, 6 gave the desired effect. It is also known that 9-f2-hydroxyethoxymethyl guanine (acyclovir) is capable of suppressing the reproduction of the herpes virus. The aim of the invention is to expand the range of substances with an improved spectrum of antiherpetic properties. The goal is achieved by the properties of a new compound of Formula 1 that is active against the herpes virus. the compound contains ribavirin nitrogenous base as a heterocycle, and the alkyl substituent acyclovir as a pentose analogue. the antiviral activity has not been previously studied. The method for preparing this compound is based on a known reaction and is carried out according to the following scheme: 0CH2Cl g-N 1 (JUCONH2 CH20CH2CH20H

the preparation is purified in two ways.

Method A. The residue is dissolved in 50 ml of ethanol, 40 cm L 40/100 µm silica gel (Chemapol. Czechoslovakia) is added (volume-wise) and evaporated to dryness. The silica gel thus treated is applied to a column (4x15 cm) with silica gel of the same degree of dispersion (the total silica gel volume is not more than 220 cm y. The elution is carried out with a mixture of chloroform ethanol (linear gradient from 0 to 30% ethanol, total volume 2 l). Fractions, containing compound I (eluting concentration of ethanol 10%, evaporated to dryness. Substance I is recrystallized from ethanol, yield 3.35 g, 45%.

Method B. The residue is dissolved by heating in ethanol, left overnight at room temperature, the precipitate is separated, the mother liquor is evaporated to dryness, the residue is recrystallized from ethanol. The yield of substances 1 3.19 g, 43%.

Both methods of isolation allow quantitative separation from the by-products.

Physico-chemical characteristics of the compound 1: t; pl.111-P2 C; (): 205 (8600), 215.3 f8600; (Sagu 118, CUIA); PMR spectrum in T) ;, 0 C (5;: m, d.): H (3) 8.18 s (W), CH2 (1), 6.01 s (2H), CHg (3) and CHa (4 3.75 s (4H) (spectrometer XL -100-15,

CUIA);

Study of the effect of drugs on the morphology of cells SU4

Found,%: C 38.61; H 5.48.

Ct} i ,.

Calculated,%: C 38.70, H 5.42.

those. R: 0.58 (.System A); 0.66 (System B); 0.57 (system VL

Systems for TLC: n butanol - water acetic acid (5: 3: 2) (A); isopropanol - ammonia - water (8: 1: 1) (B); chloroform - alcohol (7: 3) (c). Example 2. Determination of the cytotoxic effect and antiviral activity of 1-G2-hydroxyethoxymethyl -1, 2,4-triazole-5-carboxamide,

Taking into account that the claimed substance contains the nitrogenous base of ribavirin as a heterocycle, the biological activity of the new substance was studied in comparison with ribavirin

(iCN, craA).

The study of the cytotoxic effect was carried out on a transplantable cell line CUf (kidney cells of African green martghaek. Nutrient

medium No. 199 containing 103 fetal bovine serum (Gibco, Scofland).

Compound I was shown to have a significantly lower cytotoxic effect than ribavirin. Thus, at a concentration of 250 µg / ml, it practically does not change the morphology of the cells during 96 h of contact, while ribavirin at a similar concentration produces a 50% toxic effect.

Data on the effect of the claimed drug on the morphology of the cells presented in table 1.

T a b l i. c a

250

Ribavirin

125

60

250

125

Connection 1

60

1+ 2+

1 +

1+ 1 +

2+

$ 1

These data are in complete agreement with the results of studying the effect of the drug on the synthesis of cellular DNA. The determination of DNA synthesis was carried out by the inclusion of H-thymidine (α-USSR, Leningrad, sp.activate 22 curie / mmol) in the acid-insoluble fraction.

The administration of compound 1 at a concentration of 250 µg / ml resulted only in partial DNA synthesis in a CV-t cell culture in the presence of Preparations

Control

Ribavirin

(250 µg / ml) Compound 1.32x10 M (250 µg / ml

Notes: - control cells, untreated ;. Contact with H-thymidine continued for 1 hour at 37 ° C,

The study of antiviral active-30 for cytopathic effect and vyti compounds were carried out in the same cellular system, as a infectious material used herpes simplex virus type I; strain. The multiplicity of infection OjOOl 35 | ID5o on the cell. The preparations were introduced into the support medium immediately after the adsorption of the virus. Infectious virus titer determined

The effect of compound I on the development of a cytopathic effect caused by the herpes simplex virus in CVi cells

245594

inhibition of cellular DNA synthesis (44 and 49%) after 24 and 48 hours of incubation, respectively; under similar conditions, ribavirin inhibited this process by 86% (Table 2); a concentration of 125 µg / ml of compound 1 vyzgoal an unreliable decrease in the synthesis of cellular DNA, not exceeding 15%,

table 2

6.14 + 0.3

59% 0.87 + 0.3 86

44% 3.17 + 0.3 49

razhali in Rd.

A study of the effect of the test compound on the development of an infectious process in CV-i cells showed that in all concentrations used, compound I retards the development of a viral infection.

The data presented in Table. 3

T a b l and c a 3

71

A study of the effect of compound I on the accumulation of an infectious virus is presented in Table 4. Obviously, the test median has a significantly greater antiherpetic

Treatment of the herpes virus with Compound I in working concentrations hardly makes it infectious. The effect of drugs on the reproduction of the herpes virus

in cell culture Cj

Concentration

Drug mkg / ml

125

60 nome

125 data

60

thirty

15

7.5 P

eight

activity than ribavirin; a concentration of 125 µg / ml reduces the titer of the virus by 4 9 GDSA / mA and even a concentration of 7.5 µg / ml results in a significant suppression of the infectious titer of the virus, such as Stabl. thus, the test compound does not have a virucidal effect.

TableA

Suppression of virus titer, T1SCH, 5p /

Ma.

2.5 5.5 4.0 3.5 3.0 2.0 1.5 Note: The multiplicity of infection is 0.001 ID per cell. Compounds were introduced into the support medium immediately after virus adsorption. Study of; virucidal effect of compound I Variant Concentration, Virus titer, Virus-7 Virus (4 hours) - 6.5 Virus treated with compound 1 {4h, 1256.5 Virus treated with compound 1 (4h, 37 ° C) 62, 5 6.5 T a b l and c a 5 µg / ml Agtcd 50 / ml

9112455910

Thus, the test-conducted activity and selectively show that the new compound suppresses the reproduction of the viral virus has a pronounced opposite of herpes.

Claims (1)

1- (2-hydroxyethoxymethyl) -], 2,4-triazole-5-carboxyamide formula sn ₂ osn ₂ sn ₂ he with activity against the herpes virus. Zi „, 1124559 1124559