RU2009127C1 - Hydrobromide of 1-acetyl-2-isothioureido-3-indolinone protected liver against poisoning of carbon tetrachloride - Google Patents

Abstract

FIELD: medicine. SUBSTANCE: 1-acetyl-3-indoline is used as reagent 1, elementary bromine is used as reagent 2, thiourea is used as reagent 3. Reaction is carried out in the medium of dioxane. Desired product has melting point 256 C. EFFECT: improves desired product quality. 2 tbl

Description

_HBr защищающему печень от отравления четыреххлористым углеродом.The invention relates to a new compound of a number of thiocarbamide derivatives of indole, specifically to hydrobromide 1-acetyl-2-isothioureido-3-indolinone of formula I

_HBr protects the liver from carbon tetrachloride poisoning.

 This property suggests the possibility of using this compound for the prevention and treatment of acute inhalation poisoning in humans by carbon tetrachloride pairs.

 A significant increase in the use of technical fluids for various needs is accompanied by an increase in acute poisoning. Chlorinated fatty hydrocarbons have a specific cytotoxic effect, and poisoning by them often leads to death. Chlorinated hydrocarbons can be especially dangerous in emergency situations when, due to their high volatility, a large number of people can undergo inhalation poisoning.

 All patients have severe liver damage, up to fatty degeneration of the organ. The widespread use of carbon tetrachloride in industry, agriculture and military affairs makes it relevant to search for pharmacological agents that protect the body from exposure to carbon tetrachloride.

 For the situation of liver regeneration processes, derivatives of purine and pyrimidine bases are used - riboxin and potassium orotate (analogue II) (1), but their activity is insufficient.

Известно, что аналог III - наиболее активное соединение, обладающее способностью защищать печень от отравления четыреххлористым углеродом, однако его активность недостаточная (2).The closest structural analogue of the claimed compound, containing both the indole core and the thiocarbamide fragment, is N- (1-acetylindol-3-one-2-yl) thiocarbamide (analogue III).

It is known that analogue III is the most active compound with the ability to protect the liver from carbon tetrachloride poisoning, but its activity is insufficient (2).

 The aim of the invention is the creation of a new compound of a number of thiocarbamide derivatives of indole, which more effectively protects the liver from carbon tetrachloride poisoning.

 This goal is achieved by the chemical structure of a new compound of formula 1 - a thiocarbamide derivative of indole having the above property.

 The inventive compound is a white crystalline substance, hardly soluble in water, ethyl and isopropyl alcohols, readily soluble in methyl alcohol, dimethyl sulfoxide and dimethylformamide, readily soluble in ethanol when boiled. The substance is unstable to heat both in solutions and in the crystalline state. Elemental analysis and spectral characteristics are given in the example.

П р и м е р . К раствору 1 г (5,717 ммоль) 1 -ацетил-3-индолинона в 14 мл диоксана при 20^оС прибавляют раствор 0,29 мл (5,728 ммоль) брома в 14 мл диоксана, выдерживают в течение 1 мин, прибавляют приготовленный при нагревании и охлажденный до 25^оС пересыщенный раствор 0,52 г (6,84 ммоль) тиомочевины в 17 мл изопропилового спирта и кипятят с обратным холодильником 5 мин. Фильтруют в горячем состоянии, осадок промывают изопропиловым спиртом 2 х 1 мл), диоксаном (2 х 1 мл) и эфиром (4 х 2,5 мл), сушат в вакууме над фосфорным ангидридом.The inventive compound is obtained according to the following scheme. 1-Acetyl-3-indolinone is brominated with elemental bromine in dioxane followed by treatment of the resulting 1-acetyl-2-bromo-3-indolinone with thiourea.

PRI me R. To a solution of 1 g (5.717 mmol) of 1-acetyl-3-indolinone in 14 ml of dioxane at ²⁰ ° C was added 0.29 ml (5.728 mmol) of bromine in 14 ml of dioxane was allowed to stand for 1 minute, add prepared by heating and cooled ^to 25 ° C supersaturated solution of 0.52 g (6.84 mmol) thiourea in 17 ml of isopropyl alcohol and heated to reflux for 5 min. Filter while hot, the precipitate is washed with isopropyl alcohol 2 x 1 ml), dioxane (2 x 1 ml) and ether (4 x 2.5 ml), dried in vacuum over phosphoric anhydride.

1.25 g (66.4%) of 1-acetyl-2-isothiureido-3-indolinone hydrobromide are obtained in the form of grayish-white crystals with a faint pink tint, so pl. 525 ^{° C} (dec.). Under conditions of recrystallization, for example from alcohol, the substance undergoes conversion. For analytical purposes, the resulting substance was stirred with 7 ml of methyl alcohol, filtered, the precipitate was washed with methyl alcohol (2 x 2 ml. The filtrate and the washing liquid were combined, stirred with 0.38 g of activated carbon and filtered. 50 ml of ether was added to the obtained filtrate. the precipitate is filtered off, washed with ether (4 x 2 ml) and dried.

Obtain 0.5 g of the claimed compound in the form of white crystals, so pl. 256 ^{° C} (dec.). The substance is homogeneous according to TLC on silofole in UV-254.

 When mixing 5-6 mg of compound 1 with 1 ml of 5 N. caustic soda solution after 30 minutes an intense blue color occurs due to the formation of indigo (qualitative reaction).

Found,%: C 39.77, H 3.92; N, 12.89; S 10.00
C ₁₁ H ₁₁ N ₃ O ₂ ˙ HBr.

 Calculated,%: C 40.01, H 3.66; N, 12.73; S 9.71.

₁ H-NMR spectrum at D ₂ O, 100 MHz, δ ppm: 2.44 s (3H, CH ₃ ), 6.11 s (1H, H ² ), 7.15-7.61 m (4H, Ar); in DM ₃ OD: 2.42 s (3H, CH ₃ ), 6.06 s (1H, H ² ), 7.20-7.66 m (4H, Ar).

 The study of the biological activity of the claimed compound 1.

The experiments were performed on male rats weighing 140-170 g. Carbon tetrachloride was injected subcutaneously daily in a 50% molar solution for 4 consecutive days in an amount of 0.4 ml / 100 g of mass. At the same time as CCl _4, compound 1 was administered at a dose of 50 mg / kg ip. The drug was compared with the known hepatoprotector P, as well as compound III. Riboxin and potassium orotate were administered at a dose of 100 mg / kg, analog III at a dose of 50 mg / kg.

We took into account that the clinical picture of CCl ₄ poisoning develops slowly. The main symptoms of severe intoxication appear only at the end of 2 days and after 4-7 days lead to death. All patients have severe liver damage up to fatty degeneration of the organ. The protective effect of compound 1 and analogues II and III was evaluated by examining the activity of liver-specific alanine transferase (AlAT) and aspartate aminotransferase (AsAT) in blood serum. In addition, the duration of hexanal sleep was evaluated, which reflects the degree of restoration of the microsomal system of the liver metabolizing xenobiotics. A change in the activity of AlAT and AsAT in blood serum, as well as an increase in the duration of hexanal sleep, indicates the damaging effect of CCl ₄ on cell membranes and organelles of the cell. The results of the experiments are given in table. 1.

 The data obtained indicate that the use of compound 1 in 1.59 times reduces the concentration of AlAT, 1.25 times the concentration of AsAT and reduces 1.54 times the duration of hexanal sleep. Thus, in its ability to protect the liver from carbon tetrachloride poisoning, taking into account the totality of the indicators and the effect of the drug on cytolysis, compound 1 surpasses the well-known remedy II - a mixture of potassium riboxin and orotate, as well as analogue III.

 Acute toxicity was determined on white nonlinear male muscles weighing 18-22 g by a known method (3). The studied compound was administered intraperitoneally once in the form of a suspension in water with the addition of Tween-80. The group for the introduction of a single dose included 2 individuals. The total number of groups was four. Observation of the state of experimental animals was carried out for 3 days. The death of experimental animals occurred in the first 48 hours. The surviving animals then did not differ in appearance and behavior from the control.

 The toxicity of the claimed compound 1 was judged by the value of the average lethal dose calculated by the method of Prozorovsky.

 The experimental data underlying the calculation of the average flying dose are presented in table. 2.

According to the table in (3), we determine that the desired LD ₅₀ is 1030 (880-1180). Confidence limits with a 95% probability are (676-1384).

= 6,86, следовательно, уточненная средняя достоверна ( 6,86 > >2,36).t =

= 6.86, therefore, the adjusted average is reliable (6.86>> 2.36).

 The obtained result indicates that the claimed compound can be attributed to low toxicity substances, and it is less toxic than analogue III. The inventive compound I is characterized by sufficient pharmacological breadth. (56) Rychnev V.E., Frolov V.M. Regeneration stimulants in the treatment of viral hepatitis and other liver diseases. Ed. Voronezh University, 1984, p. 23.

 USSR author's certificate N 1736149, cl. C 07 D 209/36, 1990.

 Prozorovsky V. B. et al. "Express method for determining the average effective dose and its errors", Pharmacology and Toxicology, 1978, N 4, p. 494-502.

Claims (1)

1-Acetyl-2-isothioureido-3-indolinone hydrobromide of the formula

protects the liver from carbon tetrachloride poisoning.

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