KR950005303B1 - Antibiotics - Google Patents

Abstract

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Description

[Name of invention]

Sepha antibiotic

Detailed description of the invention

The present invention relates to an antimicrobial agent which is a novel cefem derivative.

Conventionally, many compounds are known as compounds having a substituted thiazozolyl acetoamide group or a substituted thiazolyl acetoamide group at the 7 position of the cepam skeleton. For example, Japanese Patent Publication Nos. 55-11,600, 55-105,689, 57-24,389, 57-81,493, 58-47,789, 58-41,887, 58-59,992, 51-149,296, 52-102,293 52-116,492, 52-125,190, 54-154,786, 57-192,394, 59 The compound of Unexamined-Japanese-Patents, such as -219,292, 60-97,982, 60-197,693, 60-231,683, is mentioned.

Patents, Japanese Patent Laid-Open Nos. 59-219,292, 60-197,693, and 60-231,683, disclose 7β-[(Z) -2- (2-aminothiazol-4-yl) -2- Methoxyiminoacetamido] -3- (1-quinuclidinio) methyl-3-cepem-4-carboxylate is described, but the compound has acute toxicity [LD ₅₀ mouse venous injection]] Is a compound that is extremely toxic even at less than 100 mg / kg and cannot be used clinically in practice.

The inventors have described 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (4-carba Moyl-1-quinuclidinio) -methyl-3-cepem-4-carboxylate and its nontoxic salts have been found to have high stability and good antimicrobial activity and have completed the present invention.

Accordingly, it is an object of the present invention to provide a novel antibacterial agent.

The present invention provides 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido represented by the following structural formula. The present invention relates to an antibacterial agent comprising (4-carbamoyl-1-quinuclidinio) methyl-3-cepem-4-carboxylate or a nontoxic salt thereof.

Examples of the non-toxic salts include pharmaceutically acceptable salts such as alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts and magnesium salts, hydrochloride salts, bromate salts, iodide salts, sulfate salts, carbonate salts and bicarbonate salts. Organic sulfonates such as acid carbonates, maleates, lactates, tartarates, methane sulfonates, benzene sulfonates, toluene sulfonates, arginine salts, lysine salts, serine salts, asparagine salts and glutamate salts. Amino acid salt, trimethylamine salt, triethylamine salt, pyridine salt, procaine salt, picoline salt, dicyclohexylamine salt, N, N'-dibenzylethylenediamine salt, N-methylglucamine salt, diethanolamine Amine salts such as salts, triethanolamine salts, tris (hydroxymethylamino) methane salts, and phenethylbenzylamine salts.

The compound of the present invention can be prepared by the method described below.

Manufacturing Method I

General formula

Wherein X is a halogen atom, the amino group and / or the compound in which the carboxyl group is protected by a protecting group, or a salt thereof

The compound represented by the above or its salt can be reacted, and if necessary, the protecting group can be removed to obtain the compound of formula (I) and its nontoxic salt.

Examples of the halogen atom of X in the general formula (II) include an iodine atom, a bromine atom, and a chlorine atom, and an iodine atom and a bromine atom are particularly preferable.

The reaction can be carried out at a reaction temperature of -10 ° -60 ° C, preferably 0 ° -40 ° C. Moreover, as a reaction solvent, anhydrous organic solvent is preferable. Examples of the organic solvent that can be used include lower alkylnitriles such as acetonitrile and propionitrile, halogenated lower alkyls such as chloromethane, methylene chloride and chloroform, ethers such as tetrahydrofuran and dioxane, and N, N-diethylformamide. Hydrocarbons such as amides such as ethyl acetate, and mixed solvents thereof.

As the protecting group in the salts of the compounds of the formulas (II) and (III), the amino groups and the carboxyl groups of the compounds of the formula (III), those which are usually used can be used.

For example, protecting groups for amino groups include formyl group, acetyl group, chloroacetyl group, dichloroacetyl group, t-butoxycarbonyl group, benzyloxycarbonyl group, trityl group, p-methoxybenzyl group, diphenylmethyl group and the like. Examples of the protecting group include p-methoxybenzyl group, p-nitrobenzyl group, t-butyl group, methyl group, 2,2,2-trichloroethyl group, diphenylmethyl group and pivaloyloxymethyl group. In addition, when silylating agents such as bis (trimethylsilyl) acetamide, N-methyl-N- (trimethylsilyl) acetamide, and N-methyl-N-trimethylsilyl-trifluoroacetamide are used, the amino and carboxyl groups are simultaneously protected. It is convenient because it can be made.

As salts of the compounds of the general formulas (II) and (III), alkali metal salts (e.g. sodium salts, potassium salts, etc.), alkaline earth metal salts (e.g. calcium salts, magnesium salts, etc.), ammonium salts, inorganic acid salts (e.g. hydrochlorides) , Bromate, sulfate, carbonate, iodide, bicarbonate, etc., organic carbonates (e.g. acetates, maleates, lactates, tartarates, etc.), organic sulfonates (e.g. methanesulfonates, benzenesulfonates, toluene Sulfonates, etc.), amine salts (e.g., trimethylamine salt, triethylamine salt, pyridine salt, procaine salt, picoline salt, dicyclonuclear amine salt, N, N'-dibenzylethylenediamine salt, N-methyl Glucamine salt, diethanolamine salt, triethanolamine salt, tris (hydroxymethylamino) methane salt, phenethylbenzylamine salt, etc.), amino acid salt (e.g. arginine salt, asparagine salt, lysine salt, glutamate salt, serine salt) Etc.) can be appropriately selected.

Removal of a protecting group can be performed by a conventional method such as hydrolysis and reduction, depending on the type of protecting group used.

Manufacturing Method II

Structural formula

A compound represented by the following formula, a compound whose -COO- group is protected with a protecting group, or a salt thereof

The compound of formula (I) and the non-toxic salt thereof can be obtained by reacting with a compound represented by the above, a compound whose amino group is protected by a protecting group, a reactive derivative in the carboxyl group thereof or a salt of the compound thereof, and removing the protecting group as necessary. have.

This method can be performed according to usual N-acylation reaction conditions. For example, in an inert solvent, it may be carried out at a temperature of -50 ° -50 ° C, suitably -20 ° -30 ° C, in the presence or absence of a base. Examples of the inert solvent include acetone, tetrahydrofuran, N, N-dimethylformamide, N, N-dimethylacetamide, dioxane, dichloromethane, chloroform, benzene, toluene, acetonitrile, or a mixed solvent thereof. Examples of the base include N, N-dimethylaniline, triethylamine, pyridine, N-methylmorpholine, and the like.

In the case of using the carboxylic acid (-COOH) represented by the structural formula (V) in the method of the present invention, for example, N, N'-dicyclohexylcarbodiimide, N, N'-diethylcarbodiimide, It is suitable to carry out the reaction in the presence of a condensing agent such as N-cyclohexyl-N'-morpholinoethylcarbodiimide, phosphite trialkyl ester, polyphosphate ethyl ester, p-toluene sulfonate and the like. In addition, when using a reactive derivative in the carboxyl group of structural formula (V), as a reactive derivative, an acid halide (for example, a chloride, an acid bromide, etc.), a symmetric acid anhydride, ethyl chlorocarbonate, trimethyl acetic acid, thioacetic acid, diphenyl Mixed acid anhydrides with carboxylic acids such as acetic acid, active esters (e.g. 2-mercaptopyridine, cyanomethanol, p-nitrophenol, 2,4-dinitrophenol, pentachlorophenol, etc.), saccharin and the like Amides and the like are used.

As the protecting group for -COO- of the compound of formula (VI), the group exemplified as the protecting group for the carboxyl group of the compound of formula (II) can be used. As the protecting group for the amino group of the compound of formula (V), a group exemplified as the protecting group for the amino group of the compound of formula (II) can be used. These protecting groups can be removed by conventional methods such as hydrolysis and reduction, depending on the type of protecting group used.

As a salt of the compound of structural formula (IV) and (V), it can select suitably from the thing illustrated as a salt of said general formula (II) and formula (III), for example.

The compound of the present invention has a strong antimicrobial activity against Gram-negative bacteria and Gram-positive bacteria, and is useful as an antibacterial agent.

When the compound of the present invention is used as an injection, it can be administered intravenously or intramuscularly by dividing 100 mg-10 g daily into 1-4 times. In addition, the dosage increases and decreases according to age and symptoms.

Injectables can be prepared by conventional methods. For example, the compound of the present invention may be dissolved in distilled water in the presence of an isotonicity agent, a dissolution aid, or the like to form an injection solution, if necessary. It may also be filled into a vial or the like in powder form to form a dissolvable injection for use. This injection is used by dissolving in distilled water for injection, physiological saline, glucose injection, amino acid sap and the like at the time of administration.

Next, 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminosaetamido] -3- (4) The data on the safety and pharmacological effects of -carbamoyl-1-quinuclidinio) methyl-3-cepem-4-carboxylate were described.

(1) mouse acute toxicity

The compound of the present invention was dissolved in physiological saline and intravenously administered to 5 ICR male and 6 week old mice. As a result, death was not confirmed even at a dose of 10 g / kg.

(2) Antibacterial activity [MIC (μg / ml)]

Staphylococcus aureus 209-P 3.13

Staphylococcus aureus E31 106 ^* 25

Eserihia, Collie NIHJ 0.1

Citrobacta and Puroindi EC-34 ^* 0.1

Ceracia Marussetsense ES-75 0.2

Pseudomonas aeruginosa EP-0.1 0.4

Pseudomonas Marudophyria EO4004 3.13

^* β-lactamase live bacteria

(3) Therapeutic Effect on Intraperitoneal Infected Mice

ICR-based, male and 4-week old mice (18-24 g body weight) were used in 10 groups. Pseudomonas aeruginosa E03235, Pseudomonas aeruginosa EO3284, and Acinetobacta powderycoaceticase E13024 were incubated at 37 ° C for 20 hours in triplicate gauze sobris to which 0.4% potassium nitrate was added. After dilution with sterile saline solution, the mixture was mixed in an equal amount with 10% gastric gumchin and 0.5 ml thereof was inoculated intraperitoneally of the mouse. One hour after the inoculation, 0.2 ml of each pharmaceutical solution was administered once under the mouse abdomen.

LD ₅₀ (mg / kg) was obtained by the Richfield Walcoxon method as the number of surviving mice 7 days after infection. CAZ (seputadidim), CFS (sepurrodin), CMX (sefamenoxime), and CTX (cell lactose core) were used as reference drugs.

The medicament of the present invention showed an excellent therapeutic effect against Pseudomonas aeruginoza and acinetobacta gircoacetigas, which are non-fermentative bacteria, compared to other medicaments.

Next, an Example and a test example are given and this invention is demonstrated in detail.

[Test Example 1]

(Production of Raw Material Compound)

7β-formamido-3- (4-carbamoyl-1-quinuclidinio) methyl-3-cepem-4-carboxylate

1.2 g of 7β-formamido-3-acetoxymethyl-3-cepem-4-carboxylic acid is suspended in 12 ml of methylene chloride, where 815 µl of N-methyl-N- (trimethylsilyl) trifluoroacetamide is added. It was added and stirred for 30 minutes. After cooling the mixture with ice, 1.25 ml of iodotrimethylsilane was added thereto, stirred for 5 minutes, allowed to come to room temperature, and stirred for another 15 minutes. The solvent was distilled off under reduced pressure, and the residue was dissolved in 12 ml of acetonitrile, and 616 mg of 4-carbamoylquinuclidin was added under ice cooling and stirred for 1 hour. 3 ml of methanol and 300 ml of ethyl ether were further added to the reaction solution, and the resultant precipitate was obtained by filtration.

This precipitate was purified by column chromatography on silica gel [developing solvent: acetone-water (7: 1) to (5: 1)] to obtain 140 mg of the target substance.

Infrared absorption spectrum (cm ^-1 , New sol): 1770.

NMR Spectrum (δ, D20): 2.30 (6H, m), 3.2-5.0 (m), 5.39 (1H, d, J = 6Hz), 5.89 (1H, d, J = 6Hz), 8.35 (1H, s)

Experimental Example 2

(Production of Raw Material Compound)

7β-amino-3- (4-carobamoyl-1-quinuclidinio) methyl-3-cepem-4-carboxylate hydrochloride

130 mg of the compound obtained in Experimental Example 1 was suspended in 5 ml of methanol, and 0.52 ml of concentrated hydrochloric acid was added at room temperature, followed by stirring for 4 hours. The reaction solution was concentrated under reduced pressure and crystallized using diethyl ether methanol to obtain 115 mg of the target substance.

Infrared absorption spectrum (cm ^-1 , New sol): 1780.

NMR spectrum (δ, D ₂ O): 2.30 (6H, m), 3.3-4.9 (m), 5.31 (1H, d, J = 6 Hz), 5.53 (1H, d, J = 6 Hz).

Example 1

(Production of the target compound)

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (4-carbamoyl-1 -Quinuclidinio) methyl-3-cepem-4-carboxylate

2- (5-Amino-1,2,4-thiadiazole)-(Z) -2-methoxyiminoacetic acid 46 mg, 1-hydroxy-1 H-benzotriazole hydrate 35 mg, N, N'- A mixture of 52 mg of dicyclohexylcarbodiimide and 1 ml of N, N-dimethylformamide was stirred at room temperature for 3 hours, then filtered and the filtrate was cooled to 0 ° C. This solution was prepared with 100 mg of 7β-amino-3- (4-carbamoyl-1-quinuclidinio) methyl-3-cepem-4-carboxylic acid hydrochloride, 2 ml of N, N-dimethylformamide and N, N-dimethyl Aniline was added to 72 μl of ice cold solution. After the mixture was stirred at room temperature for 14 hours, the reaction solution was filtered and the filtrate was added dropwise while stirring in 100 ml of ethyl ether. The precipitate that precipitated was filtered and washed with ethyl ether. 10 ml of water was added thereto, the insoluble substance was filtered off, and the filtrate was purified by reverse phase silica gel column chromatography to obtain 3 mg of the target substance.

Infrared absorption spectrum (cm <^-1> , New sol): 1775.

NMR Spectrum (δ, D ₂ O): 2.30 (6H, m), 3.1-4.0 (m), 4.16 (3H, s), 5.43 (1H, d, J = 6 Hz), 5.97 (1H, d, J = 6 Hz).

Example 2

(Production of the target compound)

7β-[(Z) -2- (5-amino-01,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (4-carbamoyl-1 -Quinuclidinio) methyl-3-cepem-4-carboxylate

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) 2-methoxyiminoacetamino] -3-acetoxymethyl-3-cepem-4- 790 mg of carboxylic acid was suspended in 10 ml of methylene chloride, and 2.1 ml of N-methyl-N- (trimethylsilyl) trifluoroacetamide was added thereto and stirred at room temperature for 1 hour. After the mixture was ice-cooled, 660 µl of iodotrimethylsilane was added thereto, the mixture was stirred for 15 minutes, and the reaction solution was concentrated under reduced pressure to obtain 7β-[(Z) -2- (5-amino-1,2, 4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3-iodomethyl-23-cefe-4-carboxylic acid was obtained. This was dissolved in 9 ml of acetonitrile, and 240 mg of 4-carbamoyl quinuclidin was added to the solution, followed by stirring for 1 hour while ice-cooling. 0.6 ml of methanol was added to the reaction solution, followed by stirring for 15 minutes. The resulting precipitate was filtered off and the precipitate was washed with acetonitrile. It was dissolved in 30% ethanol and concentrated under reduced pressure, then the residue was dissolved in acetone-water (7: 1). This solution was purified by silica gel column chromatography (developing solvents: acetone-water 7: 1 and 5: 1) to give the title compound (326 mg). The solubility of this compound in water was 30% or more, and other physical properties were consistent with those of Example 1.

Example 3

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamino] -3- (4-carbamoyl-1- 100 g of quinuclidinio) ethyl-3-cepem-4-carboxylate was dissolved in 500 ml of distilled water, and 5 ml of this solution was poured into 1 vial. It was lyophilized and used as an injection.

Claims (2)

7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamino] -3- [4-carbamoyl-1- An antibacterial agent consisting of quinuclidinio) methyl-3-cepem-4-carboxylate or a nontoxic salt thereof. 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (4-carbamoyl-1 -Quinuclidinio) methyl-3-cepem-4-carboxylate or a non-toxic salt thereof.