RU2428412C2 - Succinate of n-(2,4,6-trimethylphenyl)amide 2-(beta-n,n-diethylamino ethylamino)-4-oxo-4-(4-methyylphenyl)-2-butenoic acid - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: novel compound relates to amine derivatives of pyruvic acid amides and is biologically active - succinate of N-(2,4,6-trimethylphenyl) amide 2-(β-N,N-diethylamino ethylamino)-4-oxo-4-(4-methyl-phenyl)-2-butenoic acid of formula

.

EFFECT: compound has marked local anaesthetic and antimicrobial activity.

1 cl, 1 ex, 3 tbl

Description

The inventive compound relates to the field of organic chemistry, to amino derivatives of amides of acylpyruvic acids, and in particular to new biologically active substances - succinate N- (2,4,6-trimethylphenyl) amide 2- (β-N, N-diethylaminoethylamino) -4-oxo -4- (4-methylphenyl) 2-butenoic acid of the formula (I):

which may find application as a local anesthetic, antiseptic drug.

Known structural analogues of the claimed compounds obtained by the interaction of amides of aroylpyruvic acids with aromatic amines [A.O. Belyaev, E.N. Kozminykh, P.P. Makhmudov, V.O. Kozminykh, T.F. Odegova. Amides and hydrazides of acylpyruvic acids. Communication 10. Synthesis, antimicrobial and analgesic activity of 4-aryl-arylamino-4-oxo-2-butenoic acid arylamides // Chem.-Pharm. journal - 2004. - T.38. - No. 7. - S.19-21].

A local anesthetic drug - trimecain [Mashkovsky M.D. Medicines In 2 volumes - the 14th ed., Revised., Rev. and add. - M.: Publishing House New Wave LLC, 2000].

The aim of the invention is the synthesis of a new, not previously described succinate, N- (2,4,6-trimethylphenyl) amide 2- (β-N, N-diethylaminoethylamino) -4-oxo-4- (4-methylphenyl) 2-butenoic acid, possessing local anesthetic and antimicrobial properties. This goal is achieved by decyclization of 5- (4-methylphenyl) -2,3-dihydro-2,3-furandione with 2,4,6-trimethylaniline under mild conditions, followed by interaction with N, N-diethylaminoethylenediamine according to the scheme:

The synthesis of N- (2,4,6-dimethylphenyl) amide 2-hydroxy-4-oxo-4- (4-chlorophenyl) 2-butenoic acid (III) was carried out by analogy with the previously described procedure [Andreichikov Yu.S., Milyutin A.V., Krylova I.V., Saraeva R.F., Dormidontova E.V., Drovosekova L.P., Nazmetdinov F.Ya., Kolla V.E. Synthesis and biological activity of aroylpyruvic acid heterylamides and 5-arylpyrazole-3-carboxylic acids // Chem. journal - 1990. - T. 24. - No. 7. - S.33-35].

An example of the preparation of compound (II)

To a solution of 3.25 g (0.01 mol) of 4- (4-methyl-phenyl) -2-hydroxy-4-oxo-2-butenoic acid (III) N- (2,4,6-trimethylphenyl) amide dissolved in 15-20 ml of absolute toluene, a solution of 1.16 g (0.01 mol) of N, N-diethylaminoethylamine was added with stirring and boiled for 15 minutes. The mixture was left for 1 day at room temperature. The solution was evaporated, the resin was triturated with hexane, and the dry residue was recrystallized from cyclohexane. Received 2.53 g (60%) of crystalline substance (II) with T _PL 119-121 ° C. C ₂₆ H ₃₅ N ₃ O ₂ . PMR spectrum, δ, ppm, 0.98 t (6Н, 2СН ₂ - СН ₃ ), 2.22 s (12Н, 4СН ₃ ), 2.55 gr.s, (4Н, СН ₂ -СН ₂ ), 3.43 sq, (4H, 2 CH ₂ -CH ₃ ), 5.90 s (1H, CH), 7.05-7.74 m (7H, C ₆ H ₄ , C ₆ H ₃ ), 10.06 s (1H, CO NH ); 10.56 t (1H, NH).

An example of the preparation of the claimed compound (I)

To a solution of 4.22 g (0.01 mol) of N- (2,4,6-trimethylphenyl) amide 4- (4-methylphenyl) -2- (β, β-N, N-diethylaminoethylamino) -4-oxo- 2-butenoic acid dissolved in 15-20 ml of alcohol, while stirring, a solution of 1.18 g (0.01 mol) of succinic acid in 10-15 ml of alcohol was added, boiled for 5-7 minutes. The mixture was left for 1 day at room temperature. The solution was evaporated, the resin was triturated with hexane, the dry residue was recrystallized from isopropyl alcohol, and dried in air to constant weight. Received 4.70 g (87%) of crystalline substance (I) with T _pl. 126-128 ° C. C ₃₀ H ₄₁ N ₃ O ₆ . IR spectrum, v, cm ^-1 (crystals): 3232 (NH), 1720 ( CO OH), 1656 ( CO NH), 1608, 1596 (CO chelate, C = C). PMR spectrum, δ, ppm, CDCl ₃ : 1.13 t (6H, 2CH ₂ - CH ₃ ), 2.18 s (3H, CH ₃ ), 2.22 s (6H, 2CH ₃ ), 2 35 s (3H, CH ₃ ), 2.96 gs (8H, CH ₂ -CH ₂ ), 3.66 q (4H, 2 CH ₂ -CH ₃ ), 5.92 s (1H, CH) 6.07 s (1H, CH), 6.83 s (2H, C ₆ H ₂ ), 7.07-7.78 m (5H, C ₆ H ₄ , NH ⁺ ), 8.74 s (1H , CO NH ), 10.53 t (1H, NH), 14.34 s (1H, CO OH ).

The inventive compound (I) is a colorless crystalline substance, soluble in water, DMF, ethyl alcohol, sparingly soluble in isopropyl alcohol, insoluble in hexane.

The study of biological activity

Acute toxicity with intraperitoneal administration was determined on white non-linear male mice weighing 18-24 g. The test substance and the reference drug were injected into the abdominal cavity in the form of a solution in 1% starch mucus at the rate of 0.1 ml per 10 g of animal weight in increasing doses. The results were processed according to Prozorovsky with the calculation of the average lethal dose (LD ₅₀ ) at P <0.05 [Prozorovsky V.V., Prozorovskaya M.P., Demchenko V.M. Farmakol. toxicol., t. 41, No. 4, p. 497-502 (1978)].

The study of the compounds for the presence of local anesthetic activity. In the experiment, non-anesthetized rabbits were used - males weighing 2.0-2.5 kg. The sensitivity threshold of the cornea of the rabbit eye for tactile effects was determined. The initial sensitivity of the rabbit eye cornea (control) was determined twice with an interval of 5 minutes. A solution of the test substance in a volume of 0.4 ml was injected into the conjunctival sac of the rabbit's eye for 2 times, with an interval of 30 seconds. Each concentration of the substance was checked 8 times using the cornea of different eyes. The Rainier index was determined, which characterizes the depth of surface anesthesia and represents the sum of mechanical irritations of the cornea within an hour, and the duration of anesthesia in minutes. [Guide to the experimental (preclinical) study of new pharmacological substances. M., Ministry of Health of the Russian Federation, 2005].

The results were processed statistically using Student's criterion [Belenky M.L. Elements of a quantitative assessment of the pharmacological effect. L., 1963]. The results are considered reliable at p≤0,05. The results of the study of acute toxicity and local anesthetic activity of the claimed compounds and the comparison drug are presented in tables 1 and 2.

Table 1 Acute toxicity of compounds upon intraperitoneal administration Connections Toxicity with intraperitoneal administration, mg / kg I 255.0 (290.0 ÷ 420.0) Trimecaine 154.0 (132.0 ÷ 178.0)

table 2 The activity of compounds with surface anesthesia Connections Superficial anesthesia Rainier Index Duration of anesthesia, min I 1257.5 ± 42.6 73.8 ± 2.7 Trimecaine 559.0 ± 26.1 31.8 ± 4.4

The determination of bacteriostatic activity was carried out by the method of double serial dilutions in a liquid nutrient medium [Guide to the experimental (preclinical) study of new pharmacological substances. - M., 2000, p. 264-273]. For all test compounds, MICs were determined for pharmacopoeial strains: S. aureus ATCC 6538-P, E. coli ATCC 25922, C. albicans ATCC 885-653. Crops were made in meat and peptone broth, pH 7.0 with different concentrations of the tested compounds. The cultures were grown in test tubes on a beveled agar medium (meat peptone agar, dense Saburo medium). To determine the antimicrobial activity, 18-20-48-hour cultures of microorganisms were used. To prepare a working suspension of microbes, the grown culture was washed off with an isotonic sodium chloride solution and the microbial suspension density was set according to the turbidity standard of 5 units. Then, from the obtained microbial suspension (500 million mt / ml), a working solution of bacteria with a concentration of 5 million mt / ml was prepared. This suspension of microbes was introduced in an amount of 0.1 ml into test tubes with serial dilutions of the studied drug. Thus, the microbial load in the determination of PMA was 250,000 mt / ml. The studied compound in an amount of 0.05 g was dissolved in 5 ml of dimethyl sulfoxide, 1 ml of the obtained dilution of 1: 100 was combined with 4 ml of meat-peptone broth or Saburo liquid medium (1: 500). Next, a series of serial dilutions of the compound with a doubling concentration was prepared.

The results were taken into account after 18-20 hours of exposure of control and experimental tubes in an thermostat at a temperature of 37 ° C for bacteria and after 48 hours at 22 ° C for fungi. The minimum inhibitory concentration (MIC) was determined by the absence of signs of growth on a nutrient medium: the last tube with growth retardation (clear broth) corresponds to the MIC of the drug for this strain. The bacteriostatic effect of the studied compounds was compared with the action of dioxidine [Padeiskaya E.N. Infections and antimicrobial therapy, 2001, v. 3, No. 5, p. 150-155], fungicostatic - fluconazole [Rex J.Y., Walsh T.J., Sobel J. d / et all, Clin. Infect. Dis., 2000, v.30, No. 4, p. 662-678]. The research results are presented in table 3.

Table 3 Antimicrobial activity (μg / ml) of compound (I) Compound S. aureus E. coli FROM.
albic. I 31-62 31-62 125-250 Dioxidine 62.5-1000 3.9-62.5 - Fluconazole - - 16-32

Compound (I) has antimicrobial activity against Staphylococcus aureus and Escherichia coli (IPC 31-62 μg / ml) and yeast (IPC 125-250 μg / ml).

Thus, the succinate of N- (2,4,6-trimethylphenyl) amide 2- (β-N, N-diethylaminoethylamino) -4-oxo-4- (4-methylphenyl) -2-butenoic acid exhibits a more pronounced local anesthetic effect when less toxicity, superior to comparison drugs in terms of activity and duration of action. Therefore, the claimed compound can be used in medicine as a local anesthetic and antimicrobial drug.

Claims (1)

Succinate of N- (2,4,6-trimethylphenyl) amide 2- (β-N, N-diethylamino-ethyl amino) -4-oxo-4- (4-methylphenyl) -2-butenoic acid of the formula