RU2412177C2 - 2-hydroxy-4-oxo-4-phenyl-2-butenoate of benzothiazolylammonium, having hypoglycemic activity - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention relates to 2-hydroxy-4-oxo-4-phenyl-2-butenoate of benzothiazolylammonia, having hypoglycemic activity and formula:

.

EFFECT: high hypoglycemic activity.

1 cl, 1 tbl, 1 ex

Description

The invention relates to the field of organic chemistry, to new biologically active substances of the class 4-aryl-2-hydroxy-4-oxo-2-butenoates of heterylammonium, namely 2-hydroxy-4-oxo-4-phenyl-2-butenoate of benzothiazolylammonium ( 1) formulas

possessing hypoglycemic activity, which suggests its use in medicine as a hypoglycemic agent.

The closest analogue in structure to the claimed compound is 2-hydroxy-4-oxo-4-phenyl-2-butenoate 1,3,4-thiadiazolylammonium (2) [Synthesis and biological activity of salts of heterocyclic amines and heterylamides based on 4-aryl-2, 4-dioxobutanoic acids / N.A. Pulina., V.V. Zalesov, V.V. Yushkov [et al.] // Questions of biological, medical and pharmaceutical chemistry. - 2008. - No. 2. - S.37-40] formulas

Structural analogue 2 does not have hypoglycemic activity. Metformin (3), which is widely used in medical practice and is an analogue in action, was chosen as the reference standard [Mashkovsky M.D. Medicines - 15th ed., Rev., Rev. and add. - M .: New Wave LLC, 2005. - T.2. - S.562-563].

The objective of the invention is to search for a series of 4-aryl-2-hydroxy-4-oxo-2-butenoates of heterylammonium substances with pronounced hypoglycemic activity and low toxicity.

The problem is achieved by obtaining 2-hydroxy-4-oxo-4-phenyl-2-butenoyl benzothiazolylammonium (1), which has hypoglycemic activity.

The claimed compound 1 is synthesized by the interaction of 2-hydroxy-4-oxo-4-phenyl-2-butenoic acid (4) with 2-aminobenzothiazole in anhydrous chloroform at an equimolar ratio of reactants, at a temperature of 20-25 ° C, followed by isolation of the target product by known methods according to the scheme

An example of the preparation of compound 1. To a solution of 1.5 g (0.01 mol) of 2-amino-benzothiazole in 25 ml of anhydrous chloroform, a suspension of 1.92 g (0.01 mol) of 4-phenyl-2-hydroxy-4-oxo-2-butenoic acid is added (2) in 20 ml of the same solvent. The reaction mixture is stirred at a temperature of 20-25 ° C for two hours until the reagent is completely dissolved. The solution was incubated for 24 hours, the reaction mixture was cooled to 0 ° C, the precipitate was filtered off, recrystallized from ethyl alcohol. Yield 3.28 g (92%). Mp 160-162 ° C. Found,%: C 59.72; H 4.28; N, 8.06. C ₁₇ H ₁₄ N ₂ O ₄ S. Calculated,%: C 59.64%; H 4.12%; N, 8.18%.

IR spectrum (Specord M80, liquid paraffin, ν, cm ^-1 ): 3180ш (N ⁺ H ₃ , ОН), 1680 (С ¹ = 0), 1645, 1605 (С ⁴ = 0, C = N, С = FROM). ¹ H NMR spectrum (BS-567A (100 MHz), DMSO-d ₆ , GMDS, δ, ppm): 6.98 s (1H, CH), 7.04-7.99 m (13H, C ₆ H ₅ , C ₆ N ₄ , NH ₃ ).

The inventive compound 1 is a colorless crystalline substance, soluble in water, ethyl alcohol, dioxane, sparingly soluble in chloroform.

The hypoglycemic activity of compound 1 was studied in a model of alloxan diabetes. The experiments were performed on white outbred rats of both sexes weighing 180-200 g. Experimental hyperglycemia was modeled by subcutaneous administration of alloxan at a dose of 170 mg / kg. Compound 1 was administered orally to rats with alloxan diabetes at a dose of 50 mg / kg as a suspension in a 2% starch solution. Equivolume amount of 2% starch solution was administered to control animals. The glucose concentration in the blood of animals was determined by the glucose oxidase method [Zyuhno Z.I., Slavnov V.N., Panchenko N.I. and other Functional research methods in endocrinology. - Kiev: "Health", 1981. - S. 61-64]. Rats were deprived of food 16 hours before the experiment and for the duration of the experiment. The degree of hypoglycemic activity of compound 1 and the reference drug (metformin at a dose of 50 mg / kg) was determined 120 minutes after their administration. In addition, the toxicity of the test compound and the reference preparation were compared.

The results were processed statistically with the definition of student criterion using statistical programs Windows XP (Excel).

The acute toxicity of the claimed compound 1 when administered orally was studied on white mice of both sexes weighing 18-20 g with a determination of LD ₅₀ [Guidelines for the study of the general toxic effects of pharmacological substances // Guide to the experimental (preclinical) study of new pharmacological substances. / Under the general editorship of .-corr. RAMS, prof. R.U. Khabrieva. - 2-ed., Revised. and add. - M .: OJSC "Publishing house" Medicine ", 2005 - S.41-54]. In each series, 6 animals were used. Observation was carried out hourly during the day after the introduction of compound 1 in the form of a suspension in a 2% starch solution. The control animals were injected with an equivolume amount of 2% starch solution. The results were processed according to Prozorovsky with the calculation of the average lethal dose (LD ₅₀ ) at P = 0.05 [Prozorovsky VB, Prozorovskaya MP, Demchenko V.I. // Pharmacology and toxicology. - 1978. - No. 4. - S.497-502].

It was found that the LD _{50 of} compound 1 is 4500 mg / kg. According to the classification of toxicity of drugs, compound 1 belongs to the class of low toxic substances [Izmerov I.F., Sanotsky I.V., Sidorov K.K. Toxicometry parameters of industrial poisons. M .: Medicine, 1977, p.196-197].

Hypoglycemic effect in alloxan diabetes and acute toxicity of compound 1 Series of experiments LD ₅₀ , mg / kg Dose mg / kg Number of animals Glucose level, mmol / l 120 min Control ₁ (intact) - - 6 4.0 ± 0.1 Control ₂ (alloxan diabetes) - - 6 17.2 ± 1.3 ^{∗∗∗ 1} Compound 1 4500 fifty 6 9.9 ± 0.8 _{∗∗∗ 1 ∗∗∗ 2} Metformin 1266 fifty 6 8.4 ± 0.9 _{∗∗∗ 1 ∗∗∗ 2} (1050-1485) Notes: ^∗∗∗ = p <0.001
1 - in relation to control ₁
2 - in relation to control ₂

As can be seen from the table, compound 1 showed hypoglycemic activity in the model of alloxan diabetes and it is practically not inferior in action to metformin. In this case, the claimed substance 1 is 3.5 times less toxic than the comparison drug.

Thus, benzothiazolylammonium 2-hydroxy-4-oxo-4-phenyl-2-butenoate (1) exhibits high hypoglycemic activity and has significantly lower toxicity compared with metformin, which makes it possible to use it in medicine to create new drugs with hypoglycemic activity.

Literature

1. Synthesis and biological activity of salts of heterocyclic amines and heterylamides based on 4-aryl-2,4-dioxobutanoic acids / N.A. Pulina, V.V. Zalesov, V.V. Yushkov [et al.] // Biological Issues , medical and pharmaceutical chemistry. - 2008. - No. 2. - S. 37-40.

2. Mashkovsky M.D. Medicines - 15th ed., Rev., Rev. and add. - M .: New Wave LLC, 2005. - T.2. - S.562-563.

3. Tsyukhno Z. I., Slavnov V. N., Panchenko N. I. and other Functional research methods in endocrinology. - Kiev: "Health", 1981. - S. 61-64.

4. Guidelines for the study of the general toxic effects of pharmacological substances // Guide to the experimental (preclinical) study of new pharmacological substances / Edited by the corr. RAMS, prof. R.U. Khabrieva. - 2-ed., Revised. and add. - M .: OJSC "Publishing house" Medicine ", 2005 - S.41-54.

5. Prozorovsky V. B., Prozorovskaya M. P., Demchenko V. I. The express method for determining the average effective dose and its errors // Pharmacology and Toxicology. - 1978. - No. 4. - S.497-502.

6. Izmerov I.F., Sanotsky I.V., Sidorov K.K. Toxicometry parameters of industrial poisons. - M .: Medicine, 1977. - S.196-197.

Claims (1)

Benzothiazolylammonium 2-hydroxy-4-oxo-4-phenyl-2-butenoate of the formula:

possessing hypoglycemic activity.