SU1264839A3 - Method of producing z-2-(acyl-amino)-3-monosubstituted propeonates - Google Patents

Abstract

The compounds called (Z)-2-acylamino-3-mono substituted propionates with a general formula (I), where a meaning of the substituents is explained in the description, are the dipeptidase inhibitors which can be used in the antibacterial preparations made based on the antibiotics closed to the group of thienamycine. This patent contains a description closed to the procedures dealing with the production of these compounds.

Description

 V The invention relates to organic chemistry, namely to a method for producing g-2- (acylamino) -3-monosubstituted propeonates of the formula. . (1) where R, is hydrogen, C is C-alkyl, nat16 RJ is Cg-alkyl; R- - C-alkyl, wherein one or two hydrogen atoms in R and R can be replaced by chlorine or bromine, when R.J is a straight-line C - C-alkyl, and R is not Cj-. C.-alkyl, which can be; used in Ka4ectBe inhibitors of metabolism of peptidase. The aim of the invention is to develop an accessible method for producing Z-2- (acylamino) -3-my-substituted pro-peonates, which have the property of inhibiting the metabolism of dipeptidase in the body. Example 1. Z-2-Isovaleramido 2-butenoic acid. A solution of 1.07 g (10.5 mmol) of 2-ke of hydrobutyric acid and 0.71 g (7.0 mmol of isovaleramide in 15 ml of toluene is stirred at boiling point and water is collected in a small Dean-Stark trap. After 5 h, the solution cooled, resulting in a strong crystallization. After aging, the sludge is collected and washed with toluene, then. The yield of the white crystalline substance is 0.47 T ". 172-174 (slight pre-PL - rhythmic softening). The product is recrystallized from diisopropyl ketone TLC (4: 1 toluene-AcOH) shows only minor traces of of the isomer. The yield of white crystals was 0.32 g (25%): mp 175 (slightly pre-softening) According to given NMR, the product obtained is exclusively the Z-isomer Found: C 58.89; H, 8.55; N, 7.43 C. Calculated: C, 58.36; H, 8.16; N, 7.56. Example 2. g-2- (2,2-Dimesh1 cyclopropanecarboxamido) -2-pentenoic acid. Solution 1 , 74 g (15 mmol) of 2-keto valeric acid and 1.13 g (10 mmol of 2,2-dimethylcyclopropanecarboxamide 8392 in 20 ml of toluene are boiled with stirring, collecting water in a small Dean-Stark trap. After 20 h, the solution is cooled and treated with a weak stream of nitrogen. Before evaporation of the main part of the solvent, crystallization is induced by friction. After aging, the residue is collected on a filter and washed with toluene and some (C, Hg), jO. The yield of white crystalline product is 0.63 g (30%). T „, 154.5-155, (slight pre-softening). TLC (4: 1 toluene - AcOH) only shows extremely, insignificant traces of another isomer. NMR spectrum corresponds to the 2-configuration. Found: C, 62.86; H 8.27; N 6.75. (C ,, H ,, NO,) Calculated: C 62.53; H 8.11; N 6.63. PRI me R 3. Z-2- (3-UHKnoneHtilpropionamido) -2-butenoic acid .; A solution of 1.41 g (10 mmol) of 3-cyclopentyl propionamide and 1.53 g (15 mmol) of 2-ketobutyric acid is stirred while boiling over a small Dean-Stark trap. After 8 hours, the solution is cooled, resulting in abundant crystallization. The residue is collected on a filter and washed with toluene and CH C1. The yield of the crystalline product is 1.44 g T 180.5-182 ° C (pre-softening). The product is recrystallized from metal ethyl ketone. The yield of white needles is 0.63 g (28%) Tn 184-185 ° (insignificant.; Preliminary softening). TLC (4: 1 toluene - AcOH) shows one spot, and the NMR spectrum corresponds completely to the Z-isomer. Found: C, 63.99; H 8.67; N 6.27. (C ,, H, N03) Calculated: C 63.97; H 8.50; N 6.22. PRI me R 4. E-2- (2-Ethylhexanamido) -2-butenoic acid. 10 g of 2 ethylhexanoyl chloride is added dropwise with stirring to 25 ml of a cooled concentrated solution of NH OH, and a precipitate is immediately precipitated. The mixture is stirred for 2 h, then filtered and dried in air, resulting in a gain of 6.5 g of amide. 1.4 g (10 mmol) of the obtained compound and I, 5 g of ketone butyric acid (15 mmol) are boiled in 25 MP of toluene for 15 hours with water retention. The reaction mixture is cooled and partially evaporated in a stream of nitrogen. The product crystallizes after maturation for 3 hours. The crystals are collected, washed three times with toluene and dried in air. 1.13 (50%) of product was isolated, T-160-162, NMR spectra corresponded to a given structure and showed the presence of 5% E-isomer, TLC showed (4: 1 Tolu ol-AcOH) one spot. Found: C H 9.43; N, 5.88 (C.H.NO,) Calculated: C, 63.40; H 9.30; N 6.16. PRI me R 5. g-2- (2,2-Dimethylcyclopropanecarboxamido) -2-butenoic acid. 1.53 g (15 mmol) of 2-ketobutyric acid 1.13 g (10 mmol) of 2,2-dimethylcyclopropylcarboxamide and 20 ml of toluene are stirred at the boiling point for H h. After cooling, the crystalline product is filtered. It is washed, washed with toluene (3 x X 10 ml) and dried, resulting in a yield of 1.06 g of the product, 140-141s. those (4: 1 toluene - AcOH) show almost one spot, the NMR spectrum corresponds to the required structure. After recrystallization from Et OAc and drying, 0.533 g of the product, 142-143.5, are obtained, homogeneous by TLC. Found: C, 60.92; H 7.71; N 7.38 (C, OH.5 "Oz Calculated: C 60.90; H 7.67; N 7.10. PRI me R 6. D-2- (2,2-Dimethylcyclopropanecarboxamido) -2 - Hexanedioic acid. A mixture of 1.0 g of 2,2-dimethylcyclopropanecarboxamide, 2.4 g of 2-keto-adipic acid and 25 ml of methyl isovale; boiling for 4 hours, the water is removed using a modified Dean-Stark trap containing molecular sieve (4a). After aging at room temperature overnight, the crystalline residue is filtered off, washed with sulfuric acid and recrystallized from ethyl acetate to obtain 0.23 g of product T 163-165 °, NMR spectrum page corresponds to the required structure. Found: C 56.20; H 6.83; N 5.32. (C, iH, TNOa) Calculated: C 56.46; H 6.71; N 5.49. Example 7. g-2- (2,2-Diethyldicyclopropanecarboxamido) -2-butenoic acid. A mixture of 2.3 g of 2-ketobasic acid, 2.0 g of 2,2-diethylcyclopropanecarboxamide, and 25 ml of toluene is boiled for 16 h with removal water using a modified DiStark trap containing molecular sieves (4a). When cooled, the product does not precipitate. Sulfuric ether (25 ml) is then added and the mixture is extracted with a saturated solution of NaOHCO (three times). The combined fractions are acidified with concentrated HC1. A gummy precipitate crystallizes on mixing with water. After recrystallization from ethyl acetate, 0.31 g of product is obtained. T, 29-130C. The NMR spectrum corresponds to a product with the desired structure. Found: C, 64.01; H 8.62; N. C.2 N ,, N0. ) Calculated: C 63.98; H 8.50; N 6.22. PRI me R 8. 2- (2,2-Dimethyl-dicyclo-propan-c-amido) -2-hexenemic acid. Tertibutyl ester, D1 norleucine ester. To a suspension of 9.82 g (75 mmol) of DL norleucine in 80 ml of dioxane, placed in a 500 ml autoclave, cooled in an ice bath, was slowly (with stirring) added concentrated. Cool the mixture in an ice bath, add 80 ml of liquid isobutylene. The mixture is heated to room temperature and stirred under autogenous pressure for approximately 23 hours. After most of the isobutylene is released from the autoclave, the slightly cloudy solution is cooled in an ice bath, and then 400 ml of iH.NaOH and 500 ml. . After shaking, the layers are separated in a separate vronron; the aqueous fraction is washed with an additional 10 ml of Et, jO. The EtjO solution was shaken with 150 ml of 0.5 H.HCl. The acidic aqueous fraction is treated with 2.5 n. NaOH until strongly alkaline, and then shaken with 250 ml. EtsO solution is dried (MgSO,), filtered and

concentrated on a rotary evaporator. After prolonged vacuuming of the vessel with a high vacuum over a steam bath, a translucent, colorless oily product is obtained in an amount of 9.04 g (65%). The NMR spectrum shows the presence of diode dioxane, TLC (9: 1 CHC1 — MeOH) showed one spot.

Tertibutyl ester -N- (2, 2-dimethylcyclopropanecarbonyl) -B1-norleucine.

To a solution of 8.98 g mmol) of tertiary butyl ester of DL-norleucine and 5.05 g (50 weals) of triethylamine in 100 ml, stirred in an ice bath under a drying tube, are added dropwise (within 75 minutes) 6.39 g (48 mmol of 2,2-dimethylcyclopropanecarbonyl chloride) in 50 ml. Precipitation of Et3N. HC1 occurs during the addition of the reactant, especially towards the end. As the ice melts, the mixture warms to room temperature. After 16 h, the mixture was shaken with 200 ml of 0.5 H.HCl. CHjjCl fraction of washing with an additional 200 ml of 0.5 n.NS1, 2x200 ml of 0.5 n. NaOH and finally - 200 ml of N, the fraction is dried with MgSO4, treated with activated carbon and filtered through pelit, the filtrate is concentrated on a rotary evaporator (under high vacuum). The yield of the final product, which is a light orange substance, is 11.93 g (88%), TLC (2: 1 hexane - Et OA s) shows one spot. NMR and IR spectra correspond to the compound of a given structure. After aging for several days, the unused portion of this product crystallizes. T 52 - 65 ° C,

T-butyl 2- (2,2-dimethylcyclopropanecarboxamdo) -2-methoxyhexanoate

To a solution of 6.37 g (22.5 mmol) of N- (2,2-dimethylcyclopropanecarbonyl) -Bh-norleucine t-butyl ester in 35 ml, stirred at room temperature under a nitrogen atmosphere in the absence of light, add 2,69 ml (2, 45 g, 22.5 mmol of tert-butyl hypochlorite. After 15 minutes, a solution of sodium methoxide, obtained by dissolving 0.52 g (22.6 mmol) of sodium in 35 ml of CH „OH, is added. Stirring is continued at rooms 2683839

temperature under nitrogen in the absence of light. After 16.5 hours, the precipitated sodium chloride is filtered off. The filtrate is diluted () and washed successively with 3x50 ml of 0.5N. HC1, 50 ml of saturated

and 2x50 ml of water () phase

dried over MgSO4 and filtered. The filtrate is evaporated on a rotary

O evaporator. A pale, golden yellow oil (6.45 g) is subjected to high pressure preparative liquid chromatography, resulting in the separation and separation of 273 and 496 mg

5 two diastereomers - tert-butyl 2 (2,2-dimethylcyclopropanecarboxamides & 2-methoxyhexanoate) (corresponding to T „, .114-118 ° and 124-125.5 °), and also 1.97 g of the independent isomer (obviously Z) tertbutyl -2- (2,2-dimethylcyclopropanecarboxamido) -2-hexanoate (colorless masdo) ..

2- (2,2-Dimethylcyclopropanecarboxamido) -2-hexenoic acid.

A solution of 0.84 g (3.0 mmol) of tert-butyl-2- (2,2-dimethylcyclopropanecarboxamido) -2 hexenoate in 10 ml (CjHg), saturated with anhydrous HC1, is kept at room temperature under a drying tube. After 17 hours, the solution is evaporated and the remaining mass is dissolved in 10 ml of saturated NaHCO3. This solution is washed with an additional 15 ml of 0.5N. HC1, then dried (MgSO4), filtered and evaporated, resulting in a viscous oil. The oil is recrystallized from toluene. The yield of white crystals is 0.32 g (47%). T 119-122 °. TLC (4: 1 toluene AcOH) shows one point. The NMR spectrum confirms the formation of a practically pure Z-isomer. Note: When treating an adduct of methanol, tert-butyl 2- (2,2-dimethylcyclopropanecarboxamido) -2-methoxyhexenoate, anhydrous HC1 in the medium () O under the same conditions, the same product is formed.

PRI me R 9. (+) - g-2- (2,2-Dimesh1 Cyclopropanecarbonylamino) -2-octenoic acid, sodium salt.

Reagents - (+) - 2,2-Dimethylcyclopropanecarboxamide, 7.0 g of ethyl ester 2 - ketoooctanoic acid, 14.7 g; 50 mg of para-toluenesulfonic acid; and 100 ml of toluene are loaded into a 250 ml three-neck flask under a Dean-Stark trap containing molecular sieve tablets. The mixture was intensively boiled for 27 hours. The resulting light yellow solution was cooled and evaporated under vacuum in a water bath at 45 ° C in the presence of water to facilitate the removal of toluene. The remaining mass was suspended in 230 ml of 2H. NaOH and stirred for 3 hours. h, then the temperature is raised to and heated at it for another 2-1 / 2 h until a clear solution is formed. Then the solution is cooled, 85 ml of methylene chloride is added and the medium is adjusted to pH 8.5 with 4N HC1 with stirring. The organic layer is separated and discarded. The aqueous layer (Zbbml) was analyzed by liquid chromatography and determined to contain 37.2 mg / ml, 87% Z-isomer. Then another batch of 85 ml is added, j and the pH is adjusted to 4.5 with stirring. The organic layer is separated, and the aqueous layer is re-extracted with 50 ml of CHjCl and adjusted to pH 4.5. The combined organic extracts were dried with Na-SO., Filtered, and the i was evaporated. The resulting residue was dissolved in 150 ml of isopropanol and 15 ml of water and the pH was adjusted to 8.2 with 2 n. NaOH. The resulting solution is concentrated to an oily residue, which is rolled with isopropanol until it becomes a crystalline substance, which indicates that most of the water has been removed. It is recrystallized from 120 ml of isopropanol (cooled on ice for 1 hour), filtered, washed with 50 ml of cold isopropyl, then with a large amount of acetone, dried at (0.1 mm) for 2 hours, resulting in 10 74 g (63.2%) of crystalline material, characterized by a single peak-chromatograph, T Т d 241-243 43. These solutions are poured all together with stirring. The resulting crystalline mass is heated and two transparent layers are obtained, again intensively mixed with cooling and a crystalline product is obtained. This product is allowed to mature at room temperature for two days. It is then filtered, washed with 2x10 ml of water and 2x10 ml of 50% methanol and dried under vacuum. The yield of crude quinine salt is 44.8 g (48.7%), the yield of monohydrate, T: 113-116 ° C, 94.4, C 1.0; СНС1д. This material is recrystallized from acetone and get 24,35 gprodukt, T 127-130 ° C. The purified quinone salt is converted into an acid by treatment with an aqueous base and chloroform, and subsequent treatment with an acid, to obtain (96%) 3.9 g of the product having Cot-1 146.0 ° as a result. The acid is converted to amide as follows. 30.5 g of (+) acid are added through a dropping funnel over a period of 5-10 minutes to 54 ml of oxalyl chloride, containing 1 drop of dimethylformamide, via a dropping funnel. The mixture is stirred overnight at room temperature. A prostate solution is formed which is added to 100 ml of methylene chloride for dilution. The excess oxalyl chloride is removed by concentration, and the mixture is repeated twice with methylene chloride. The resulting solution is diluted with an equal volume of methylene chloride and continuously added via a droplet) funnel to about 100 ml of anhydrous liquid ammonia, which is diluted with 100 ml of methylene chloride. Example 10. D-2- (2,2-Dichlorocyclopropanecarboxamido) -2-butenoic acid. 2,2-Dichlorocyclopropanecarboxamide. 7.1 g of a sample of 2,2-dichlorocyclopropeicarbonyl chloride is added dropwise to 75 ml of concentrated ammonium hydroxide with stirring intensity. The temperature of the reaction mixture is maintained below 10 ° C in an ice bath. The mixture is stirred in an ice bath for 30 minutes, then for 1 h at room temperature. Aqueous ammonia is evaporated under reduced pressure (ban at 50G). The remaining residue is extracted with hot thylacidate (3x30 ml). The extracts are boiled down to a volume of 40 ml and 20 ml of hexane is added to them. After cooling on ice, the solid residue is filtered off, the mixture is washed with ethyl acetate: hexane (1: 1) and dried, resulting in 2.7 g of 2.2-dipropyclopropanecarboxamide, Т „„ 14491 146. The NQR spectrum corresponds to the compound of the given structure. Found:. C, 31.26; H 3.31; N 9, 45.79. (С N0) Calculated: С 31,20; H 3.27; N 9.10; Cl 46.04. Another 1.3 g of amide, T „143-145 °, can be recovered from the mother liquor, Z -2- (2,2-Dichlorocyclopropanecarboxamido -2-butenoic acid. A mixture of 1.53 g (15 mmol) of 2-ketoic acid , 1.54 g (10 mmol) of 2,2 dichlorocyclopropanecarboxamide and 10 ml of toluene are boiled for 12 h with removal of water using a modified Dean-Stark trap containing molecular sieves (4a). Add an additional 0.7 g 2-ketobutyric acid and the reaction mixture is boiled for a further 12 hours. The mixture is cooled, diluted with 20 ml of toluene and extracted with an unsaturated sodium bicarbonate solution. (3x10 ml). The extracts are combined, washed with sulfuric ether and acidified to pH 3 (pH meter) with concentrated hydrochloric acid. A resin-like substance precipitates out, which soon hardens. It is filtered, washed with water, dried and recrystallized from nitromethane , resulting in a yield of 423 mg of Z-2- (2.2 dichlorocyclopropanecarboxamido) -.2-butenoic acid, T = 188-189.5 ° C. The NMR spectrum corresponds to the compound with the desired structure. Found: C, 40.48; H 3.80; N 5.91; C1 29.53. (CgHgCl NO,) Calculated: C 40.36; H 3.81; N 5.88; From 29.78. Example 11. g-2- (2,2-Dichlorocyclopropanecarboxamido) -2-octenoic acid. A mixture of 1.19 g (7.5 mmol) of 2-keto acid, 0.77 g (5.0 mmol) of 2,2-dichlorocyclopropanecarboxamide, and 5 ml of toluene is treated and reacted in the same manner as Example 10. Crude product (537 mg ) purified by converting it to methyl ester (BF / CHjOH), preparative TLC (boiled G, 4, 1 hexane: EtOAc) and om, to pure Z-methyl ester (0.3 M Li OH / CH OH), whereby 88 mg of Z-2 (2,2-dichlorocyclopropanecarboxamido) 2-octenoic acid are obtained in the form of a partially 39 retinal resin. NMR spectrum (DM50-a: 5 9.68 (S, iH, NH); 6.50 S (t, IH H); 2.835 (t, Sh. Jblfff (vjj, ub g, 0.87 Mt, ЗН, CH) .Example 12, Z-8-Bromo-2- (2.2Imethylcyclopropanecarboxamido) -2-octenoic acid. To a suspension of 14.4 g (0.3 mol); ..: 50% dispersion of NaOH in 360 ml toluene, cooled in an ice bath, under nitrogen for 45 minutes, a solution of 146 g (0.6 mol) of 1,6-dibromhexane and 57.6 g (0.3 mol) of ethyl 1, 3- dithian-2-carboxylate in 120 MP of DMF. The cooling bath is removed and the mixture is stirred at room temperature for 2 hours. The reaction mixture is dried with water (3x210 ml), dried over MgSO4 and evaporated under reduced pressure. resulting in 179.5 g of a yellow oily product containing the desired anhydrous dithian, 1,6-dibromhexane, and mineral oil. This crude product is used to carry out the next reaction without purification. To a suspension of 426 g (2.4 mol) N bromosuccinamide in 800 a ml of acetonitrile and 200 ml of water was added over 45 minutes a solution of crude dithian in 100 MP acetonitrile was added. The temperature of the reaction mixture is kept below stirring for 10 minutes. The dark red reaction mixture is poured into 2 liters of hexane CHjCl (1: 1). The solution is shaken with a saturated solution of Na HSO (2 x X 400 ml) and water (1 x 500 ml). Then, 400 K of a saturated solution of VA is added in small portions to the solution (intensive release of CO). After the foaming is complete, the funnel is dried for three days and the aqueous layer is separated. The organic solution is extracted with a saturated solution of Na COj (400 ml) and water (500 ml) and dried over MgSO4. After removal of the solvent under reduced pressure, 133.8 g of crude I-6-dibromhexane and mineral oil are obtained. This crude product is used without purification for the next reaction. A mixture of 133.8 g of crude bromo ketone ester, 133 ml of 50% hydrobromic acid and 267 ml of acetic acid is heated at 90 ° C (internal temperature) for 75 minutes. The dark solution is evaporated under reduced pressure until until the main part of the acetic acid is removed. The residue was dissolved in 500 ml of sulfuric ether, washed with water (2 x 100 ml) and extracted with a saturated NaHCOj solution (3 x 200 ml). The combined NaHCOj extracts are extracted with ether (2x100 ml) and acidified with concentrated HC1. The resulting oil is extracted with ether, (3x200 ml). The ether extracts are washed with water (1x100 ml) and the liquors (1x100 mp) are dried over MgSO4. After removal of the sulfuric ether under reduced pressure, 46.2 g of pure bromo-keto acid homogeneous according to those obtained (silica gel, 4: 1 toluene — acetic acid) are obtained. The NMR spectrum corresponds to the compound of the desired structure. A mixture of 46.1 g (0.194 mol) of bromo keto acid, 17.6 g (0.156 mol) of 2,2 dimethylcyclopropanecarboxamide, and 450 ml of toluene is heated at boiling point for 13 h, collecting water in a small Dean-Stark trap. After cooling, the clear reaction mixture is extracted with a saturated solution of NaHCOj (4x100 ml), the combined extracts are washed with sulfuric ether (2x100 ml) and then brought to 3.5 (pH meter) by addition of concentrated HCl. The resulting BCKopg oil crystallizes. The residue is filtered off, washed well with water and dried. After recrystallization from acetonitrile, 22.5 g of g-8-bromo-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid are obtained. T „d 151-153С homogeneous according to those (4: 1 toluene-acetic acid). The NMR spectrum corresponds to the compound of the desired structure. Found: C 50.66; H 6.96; N 4.45; Br 23.95. (C, H ,, BrNOp Calculated: C 50.61; H 6.67; N 4.22; Br 24.05. Based on this procedure, the following α-bromo compound was obtained: d-6-bromo-2- ( 2,2-dimethylcyclopro: pancarboxamido) hexeneic acid; Z-7-6pOM-2- (2,2-dimethylcyclopropanecarboxamido) -heptenic acid; Z-9-6pOM-2- (2,2-dimeTylcyclopropanecarboxamido) -2-nonenoic acid ; Z-1O-bromo-2- (2,2-dimethylcyclopropanecarboxamic) - decenic acid; Z-8-6pOM-2- (2,2-dimethylcyclopropanecarboxamido) -octeneic acid. The study of the location of thienamycin, its natural analogs and semi-synthetic derivatives allowed you are based on obvious metabolic cleavage by eluting in various studied species of living organisms (mice, rats, dogs, chimpanzees, rhesus monkeys). The degree of metabolism can be judged by low urinary excretion and short half-lives of plasma. by the action of renal dipeptidase (E.C.3.4.13.11) .In order to demonstrate the ability of the compounds of formula 1 to inhibit the action of the renal enzyme dipeptidase, an in vitro method was used. It measures the ability of compounds to inhibit the hydrolysis of glycyl dehydrophenylalanine (HDP) using a solubilized dipeptidase isolated from porcine kidney. The technique is as follows: To 1 ml of a system containing 50 mm MOPS (3-N morpholino propanesulfonic acid) buffer, pH 7.1, 6 mg of the lyophilized enzyme is added and the test compound at a final concentration of 0.1 mm. After 5 minutes of incubation in a thermostat at 37 ° C, HDF is added at a final concentration of 0.05 mm. The thermostatting is continued for another 10 min at 37 ° C and the hydrolysis of HDF is measured by the change in optical density with time at 275 im. The inhibition of enzyme action is determined by comparison with the results of a control experiment carried out without the Ki inhibitor. This is the inhibitor concentration at which 50% inhibition of the enzyme is achieved. HDF is used as a substrate in this method compared to thienamycin, since it has a significantly higher maximum rate of hydrolysis of renal dipeptidase, which reduces the amount of enzyme required. Both HDF and thienamycin have the same similarity to renal dipeptidase, and Ki tested inhibitors were found to be identical for the two substrates. In addition to this in vitro experiment, an i.n vivo experiment was conducted to measure the ability of test compounds to inhibit metabolism to increase the yield of thienamycin in the urine of mice. The method involves co-administration of the test compound intravenously or subcutaneously at a dose of 10-100 mg / kg with thienamycin in an amount of 10 mg / kg. Then compare the content of thienamycin in the urine over a period of 4 hours and its content in the urine of the control group of animals that were not injected with the test compound. The urinary thienamycin content is in all cases measured using a cylindrical or disk diffusion cell. This bioassay with Stapky lococcus aureus ATCC 6538 as test organisms has a sensitivity range from 0.04 to 3.0 g / m. The results of an in vitro experiment.

(Z) - 2-Acylamine-2-butene acids

C02H

sns

Table I o Use 1 ml of the system 50 mm MOPS buffer, pH 1.7. To this is added 5 g of porcine renal enzyme and an amount of the test compound such that its final cohort is 0.1 mn. After 5 minutes of incubation at 37 ° C, the amount of HDF is added so that its final concentration is 0.05 mM. The system is again thermostatic for 10 min. Hydrolysis of HDF is determined by the change in its optical density with time at 275 im. Enzyme inhibition is determined by comparison with the results of a control experiment conducted in the absence of an inhibitor, and expressed as a percentage of inhibition. Ki is an indication of the inhibitor concentration required to achieve 50% inhibition of the enzyme. This is a calculated value resulting from several in vitro samples at concentrations that lead to inhibition below or above the 50% inhibition point. The results are presented in table 1.

CH, CHj -CHjCH (

(СН,) ЗСН I | Д I 3 I fCHzHQ 176-177VIII (С1% г-О 180.5-181VII нн, 144-145IV 167-168V

142-143,5 XII

45.5-146,5 IV

145.5-146,5Y IV

201-202IV

131-132 IV

183-184IV

s

129-130IV

189-190,5 XII

188-189,5 V

191-192V 1264839 29 "O 3 5

27 C H N0,

C, H, N 4

10 15 3

.И.5 «0з

C, H, N 19

19.8

53 С, Н N0

C, H, N (10)

tch tg 3

C, H, N45

12 С Н N0

11 f7 3

22 Ch, I., “Oz

C, H, N3

ten ,

C, H.N 86

32 С Н N0,

C, H, N1,6

10 13 3

C, H, N, Cl 08

18 C N SSO 893

C, H, N, Br 03

23 CgHgBrNO 18 Continuation of Table 1 C, H, N25 p, “. C, H, N (54) St “.,“ H, N, C (16) N0 C, H, N12,

.СНз

172-174,5 XII

The results of the in vivot in vivo experiment were tested in mice as follows. Mice weighing 20 g, female M25 (Diabetes Charles River) were subcutaneously injected with a selected dose of chemical inhibitor. After about 2 minutes, a certain dose of thienamycin is administered intravenously. The control with thienami-jg qing is also carried out. The level of thienamycin in the urine, expressed as a percentage of the administered dose, is measured using a biochemical test. The results are presented in table 2. Two numbers of the studied compounds coincide with the numbers of the compounds from table. 1. Compound 7 is 2-isovaleram-2-butenoic acid, compound 10 is g-2-cyclopropylcarboxamido-2-butenoic acid. Compounds of 2-isovaleramide-2-butenoic acid, compound 7, Z-2 (2,2-dimethylcyclopropanecarboxamido) Effect of co-administered 2-nd acid (compound 7) or no urine of mice, individuals, SL, weight 20 g

BB ILIpkzh

pkzh pkzh

Continuation of table 1

4.7

19 C.

C.H.N

10 10

30jt5 33 2-butenoic acid were studied in more detail in experiments in viVo in combination with thienamycium (TM) in experiments on mice. The general test procedure is similar to that described in Example 2. The results obtained are presented in Tables 3 and 4. tl Control. Table 3 vaperamidobutenova d thienamycin in the Charles River,

(b) Entered together.

Influenced in conjunction with Z-2- (2,2-dimesh1Cyclopropanecarboxamido) butenoic acid (compound 2) on the output of thienamycin in the urine of mice. Mice, females, SL Charles River, weighing 20 g.

(b) Co-administration.

: UNIT ".,

, 2

: TMtlOO mg / kg inhibitor 0.06

In another experiment on mice, systemically in three raes by jointly veterans antibacterial action of dyne with 2-isovaleramido-2-butene t enamshdan is increased by approximate acid.

Table4 231264 The effect of co-administered 2-invaramido-2-butenoic acid on the systematic efficacy of thienamycin in the treatment of Staphylococcus aureus infection.5 Male dogs were used to study the effect of dipeptidase inhibitors on urinary N-formimidoyl thienamycin in the urine. In the control experiment, the dog was injected with 5 mg / kg IV N-Yu formimidoyl thienamycin without inhibitor. In the second O1tyte use the same amount of N-formimidoylthienes in the urine N-forms (5 mg) kg (IV) through 3 or in the dog's body. The studied compound

Y-Formimidoylthienamycin

plus Z -2-isovaleramido2-butenoic acid

shs z-2- (2,2-dimesh1cyclopropanecarboxamido) 2-butenoic acid. Thus, the resulting compounds-35 and the general formula 1 possess the property of inhibiting inventively 839 24 qin and also introducing z-2-isovaleramido-2-butenoic acid to three doses, each pSg 20 mg / kg of the test compound. N-doses were administered immediately after the injection of N-formimidoylthienamycin, the second at the 40th minute and the third at the 60th minute. In the third trial, one 2 mg / mg g-2- (2,2-dimethylcyclopropanecarboxamido) -2-butenoic acid is used, administered just prior to the injection of N formimidoylthienamycin. The results obtained are presented in Table 5. Table 3 milking up after its introduction: Exit in 1 urine,% metabolism of dipeptidase and can be used in medicine.

Claims (1)

METHOD FOR PRODUCING g-2- (ACIPAMINO) -3-MONOSubstituted PROPEONATES of the general formula 1 RjCONH · ^ / А С = С R ^ OOC'- where R ₁ is hydrogen, C ₁ is C _g -alkyl, sodium; R ₂ —C ₃ —Cd-alkyl; R ₃ - C ₄ - _{1 °} C -alkyl, wherein one or two hydrogen atoms in R ₂ and R ₃ may be substituted by chlorine or bromine when R ₃ - straight C ₁ - C ₄ alkyl, a Rj is not C ₃ ~ C ₄ -alkyl, characterized in that the compound of general formula II ABOUT R CH _g COOR ,,, where R ₁ and R ₃ have the indicated meanings, are reacted with an amide of the general formula III ABOUT II R ₂ cnh ₂ , where R ₂ has the indicated value, in the solvent medium at its boiling point and the process is carried out at a mass ratio of compounds of the formula ΪΙ and III (1-4): 1 with simultaneous azeotropic distillation of the resulting water. SU < _w 1264839> O ' I 264839