CS199295B2 - Process for preparing new 7-n-acylamino-alpha-aryl or thienylacetamido-3-/heterocyclothiomethyl/-3-cephem-4-carboxylic acids - Google Patents

Description

The invention relates to a process for the preparation of novel cephalosporins which can be used as active substances, in particular against the genus Pseudomonas, otherwise they usually have a broad spectrum of activity.

It is known that cephalosporin compounds such as cephalotin and cefazoline are very effective against a wide range of Gram positive and Gram negative bacteria.

However, these substances have no effect against Psudomonas aeruginosa infections, which have been increasing recently and which are often very difficult to treat. Cephalosporins active against Pseudomonas aeruginosa are not yet available.

It has now been found that certain cephalosporin derivatives of the same broad spectrum as other substances of this type and their pharmaceutically acceptable salts also have an effect against Pseudomonas aeruginosa and can be used to prevent and treat diseases caused by the microorganism.

These derivatives also have high activity against other microorganisms against which conventional cephalosporins are poorly active. Thus, it is highly effective against Pseudomonas aeruginosis, indolpositive Proteus, Serratia, Enterobacter aercgenus, and cephaloridine resistant Escherlchia coli.

Accordingly, the present invention provides a process for the preparation of novel 7- [N-acylamino-α-aryl- or thienyl-lacetamido] -3- (heterocyclothiomethyl) -3-cephem-4-carboxylic acids of the general formula I ho-a-conh-ch-conh

R

CH 2 -Ua-t mnu where

Λ means a ninnocyclic or polycyclic heteroaromatic radical containing at least 1 nitrogen atom as a heteroatom selected from the group consisting of naphthyridine, pyridopyrimidine, pyridine, pyrimidine or piridazine, optionally substituted by 1

A 193295 substituent selected from the group consisting of alkyl of 1 to 4 carbon atoms, or alkylthio of 1 to 4 carbon atoms,

R is a phenyl radical optionally substituted by 1 or 2 substituents from the group hydroxyl, amino or thienyl, and

Het means a tetrazolopyridazine residue, a triazolopyridazine residue or a triazolcipyridine residue, as well as non-toxic pharmaceutically acceptable salts of these compounds, characterized in that the compound of formula II is reacted

HO — A — COOH (II) where

A is as defined above, or a reactive derivative thereof with a compound of formula III

H.N-CH-CONH-, o (III)

COOH where

R and Het are as hereinbefore defined and the compound prepared is optionally converted to its non-toxic, pharmaceutically acceptable salt.

The reaction may be carried out in a solvent such as polar solvents such as dichloromethane, chloroform, acetone, tetrahydrofuran, dioxane, acetonitrile, methyl isobutyl ketone, ethanol, dimethylformamide, dimethylacetamide, dimethylsulfoxide, nitromethane, hexamethylphosphoric triamide, sulfolane and the like. toluene, petroleum ether, n-hexane and the like. Mixtures of these solvents with one another or mixtures thereof with water may also be used.

Reactive derivatives of the compound of formula (II) are carboxyl group derivatives, for example halogens, anhydride, azolide, active ester, azide and the like. Examples of such derivatives are mixed anhydrides or symmetrical anhydrides with acids such as dialkylphosphoric, phenylphosphoric, diphenylphosphoric, dibenzylphosphoric. suitable are halogenated phosphoric, dialkylphosphoric, sulfuric, methanesulfonic, toluenesulfonic, naphthalenesulfonic acids, furthermore alkyl carbonates, aliphatic carboxylic acids, e.g. tetrazole and the like and the active esters such as cyanomethyl, methoxymethyl, vinyl, propargyl, p-nitrophenyl, 2,4-dinitrophenyl, trichlorophenyl, pentachlorophenyl, methanesulfonylphenyl, phenylazophenyl, phenylthio phenyl ester, p-nitrophenylthioester, p-cresolthioester, carboxymethylthioester, pyranyl ester, pyridylester, piperidyl ester, 8-quinolylthioester and esters of N, N -dimethylhydroxylamine, 1-hydroxy-2- (1H) -pyridone, N-hydroxysuccinimide or N-hydroxyphthalimide.

When the compounds of formula (II) are used in the form of the free acid or a salt thereof, it is preferable to carry out the reaction in the presence of agents which facilitate it. Such agents are, for example

N, N‘-dicyclohexylcarbodiimide,

N-cyclohexyl-N-morpholinethylcarbodiimide, N-cyclohexyl-N- (4-diethylaminocyclohexyl) carbodiimide,

N-N‘-diethylcarbodiimide,

N, N‘-diisopropylcarbodiimide,

N, N‘-carbonyldi (2-methylimidazole), N-ethyl-N- (3-dimethylaminopropyl) carbodimide, pentamethylenketene-N-cyclohexylimine, diphenylketene-N-cyclohexylimine, alkoxyacetylenes,

1-alkoxy-1-chlorethylenes, trialkylphosphites, polyphosphoric acids in the form of ethyl ester, isopropyl ester of polyphosphoric acid, phosphorus oxychloride, oxalyl chloride, triphenylphosphine, diethylphosphonylazide, diphenylphosphonylazide,

2-ethyl-7-hydroxybenzisoxazolium salts, 2-ethyl-5- (m-sulfonyl) isoxazolium inner salts with hydroxide, chloromethylene) dimethylammonium chloride and the like.

Examples of salts of compounds of formula III are alkali metal or alkaline earth metal salts, for example sodium, potassium, calcium and the like, salts with organic amines such as trimethylamine, triethylamine, quinoline, conidin and the like and with organic sulfonic acids such as toluenesulfonic acid, naphthalenesulfonic acid , tetralin sulfone, trifluoroacetic acid, hydrochloric acid and the like.

The compounds of formula (I) are valuable antibacterial agents and can be used as additives to livestock feed, poultry and livestock medicaments as well as in human medicine. These agents are particularly suitable for the treatment of infectious diseases caused by Gram-positive bacteria such as

Staphylococcus aureus,

Staphylococcus epidermidis,

Staphylococcus pyogenes,

Diplococcus pneumoniae,

Sarcina lutea,

Bacillus subtilis,

Clostridium perfringens a

CoTynebacterium diphtheriae and gram-negative bacteria such as Escherichia coli,

Neisseria genorrhoeae,

Salmonella typhi,

Klebsiella pneumoniae,

Shigella dysenteriae,

Shigella flexneri,

Shigella sonnei,

Enterobacter aerogenes,

Próteus mirabilis,

Próteus vulgaris,

Pseudomonas aeruginosa and Serratia marcescens.

For this purpose, the compounds according to the invention are used singly or in admixture with pharmaceutically acceptable carriers or diluents or with other active substances intramuscularly or intravenously.

The dosage of the compounds of the formula I depends on the weight, age and conditions of administration, on the type of bacterium and on the pharmacological properties of the selected substance. In general, a dose of 2 to 400 mg / kg body weight per day, preferably 8 to 120 milligrams / kg body weight per day, in a single dose or 1 to 5X daily is used for intramuscular or intravenous administration of the compounds of formula (I).

For intramuscular or intravenous administration, the compounds of the invention are used in the form of sterile solutions or suspensions containing a pharmaceutical diluent or carrier such as water, saline, Ringer's solution, glycerin, polyethylene glycol and the like. These pharmaceutical preparations may contain other excipients, emulsifiers, topical anesthetics or salts for adjusting the osmotic pressure. The compounds of the invention may also be applied topically in the form of an ointment or cream to the skin or other organs for sterilization or disinfection.

The invention will be illustrated by the following examples and comparative examples, which are not intended to limit the invention. All parts, percentages and ratios are by weight unless otherwise indicated.

Example 1

Preparation of 7- [D-α- (4-hydroxypyridine-3-carboxamido) -α-p-hydroxyphenylacetamido-3-phthrazolo [4,5-b] pyridazin-6-ylthiomethyl] -3-cephem-4-carboxylic acid

OH

N '

To a solution of 9 ml of dimethylsulfoxide is added 0.606 g of triethylamine and 0.472 g of 4-hydroxypyridine-3-carboxylic acid N-hydroxysuccinimide ester, 1.25 g of trifluoroacetic acid in the form of 7- (D -? - amino-.alpha.-hydroxyphenylacetamido). 3- (tetrazolo [4,5-b] pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid, and the mixture was allowed to react for 12 minutes at room temperature of 20-30 ° C. The solution was then added dropwise to 200 mL of acetone and 100 ml of diethyl ether was added to give crystals which were collected by filtration to give 0.99 g (yield 70.2%) of the title compound as a triethylamine salt.

This triethylamine salt was dissolved in 10 ml of water, the solution was adjusted to pH 3 with 2 N hydrochloric acid with stirring and ice-cooling. 15 ml of water and 6 ml of methanol are added to the gelled crystals to crystallize. The crystals were filtered, washed with methanol and dried over phosphorus pentoxide under reduced pressure to give 0.84 g of the title compound.

This material was dissolved in 8 ml of dimethylsulfoxide and 0.26 g of sodium 2-ethylhexanoate was added, followed by stirring at room temperature for 20 minutes. The solution was then added dropwise to 200 ml of acetone. The formed crystals were filtered off, washed with acetone and diethyl ether and dried over phosphorus pentoxide under reduced pressure. 0.85 g of product is obtained in the form of the sodium salt.

Melting point: 253-257 ° C with decomposition.

139295

Absorption spectrum in IC light: _pn- ¹ in _nuj1 = 1760, 1660, 1630, 1610. Example 2

Using the method of Example 1, 7- [D-α- (4-hydroxy-1,5-naphthyridine-3-carboxamido) -α-hydroxyphenylacetamido] -3- (pyrido [2,1-c] 1-s-triazol-3-ylthiomethyl) -3-cephem-4-carboxylic acid

mp 221-224 ° C with decomposition.

Example 3

In a similar manner to Example 1, using the appropriate starting materials, 7- (Da- (4-hydroxypyridine-3-carboxamido) -α-hydroxyphenylacetamido] -3- (tetrazolo [4,5-b] pyridazine-6-) can be obtained ylthiomethyl) -3-cephem-4-carboxylic acid

OH

X ^ yCONH-CH-CONH -] -

COOH J, m.p. 253-257 ° C.

Other compounds listed in the following table can be obtained as described above.

HO — A-Het

Melting point ° C (dec.)

254 to 262

242 to 251

261 to 270

263 to 272

HO — A—

R — Het

Melting point ° C (dec.)

253 to 261

251 to 261

248 to 255

245 to 253

252 to 259

242 to 250

250 to 257> 270

248 to 256

253 to 260

The minimum inhibitory concentration has the results shown in the following table. Minimum inhibitory concentration (jng / ml)

Microorganism Example compound

122 CET * CEZ **

Staphylococcus 0.2 1.56 0.05 0.1 0.2 Aureus 209P Escherichia 0.78 0.78 3.13 12.5 1.56 coli NIHJ Klebsiella 0.78 3.13 6.25 3.13 1.56 pneumoniae 602 Pseudomonas 0.78 3.13 3.13 > 200 > 200

* CET (cephalotin)

ΓΧ ^ S ^ CHtCONHCH £COCH _i

COONa.

** CEZ (cefazoline)

N = N | N-CH 2 CONHO

COON.

The following comparative examples illustrate processes for the preparation of the starting materials used in the preparation of the compounds of the invention. Other starting materials can be prepared in a similar manner.

Comparative Example 1

Method for the preparation of 7-amino-3-tetrazolo [4,5-b] pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid

N

2.72 g of 7-aminocephalosporanic acid are suspended in 16 ml of water and 8 ml of acetone and a suspension of 2.1 g of sodium carbonate in 16 ml of water are added dropwise over 7 minutes. The internal temperature of the reaction mixture was then raised to 50 ° C, then a suspension of 2.3 g of 6-mercaptotetrazolo [4,5-b] pyridazine in 35 ml of acetone was added dropwise over 10 minutes. The resulting mixture was heated at reflux for 6 hours, then allowed to stand at an internal temperature of 61 ° C. The solution had a pH of 8, then the mixture was adjusted to pH 3.6 by addition of 6 N hydrochloric acid under ice-cooling, and the pH was measured with a pH meter, followed by stirring for one hour. The precipitated crystals were collected by filtration and dried over phosphorus pentoxide under reduced pressure to give 2.39 g of the title compound, m.p. 202 DEG-205 DEG C. with decomposition.

Comparative Example 2

A process for preparing 7-amino-3- (s-triazolo [4,5-b] pyridine pyridazin-6-yl-thiomethyl) -3-cephem-4-carboxylic acid «? 'Η r ^of>

CH ^ OCOCH,

COOH

2.72 g (0.01 mol) of 7-aininocephalosporanic acid are suspended in 20 ml of water and 10 ml of acetone, and then a suspension of 1.89 g of sodium hydrogen carbonate in 10 ml of water is added dropwise over 10 minutes. The internal temperature of the reaction mixture was then raised to 50 ° C and a suspension of 1.9 g (0.0125 mol) of 1,2,3-triazolo-7, O-pyridazin-6-ylthiol in 20 ml of acetone was added over 8 minutes drop by drop. The resulting mixture was heated at reflux for 4 hours at a constant internal temperature of 63 ° C and then cooled with ice. When the internal temperature of the reaction mixture reached 3-4 ° C, the pH of the mixture was adjusted to 5 by adding 6 N hydrochloric acid under ice-cooling, followed by stirring for 10 minutes, bringing the pH to 3.6. After stirring for one hour, the precipitated crystals were collected by filtration, washed with 10 ml of water and twice with 10 ml of acetone and dried over phosphorus pentoxide under reduced pressure to give 2.58 g of the title compound in a yield of 72.5%. for decomposition.

Cleaning

A suspension of 1 g of the crude product thus obtained in 10 ml of water is stirred under ice-cooling, and 9 ml of 12 N hydrochloric acid are added dropwise over 10 minutes. After stirring for 20 minutes, the insoluble material was removed by filtration and the mother liquor was adjusted to pH 3.6 by addition of 7 ml of 30% sodium hydroxide and 3 ml of 10% sodium hydroxide. After stirring for one hour, the resulting crystalline precipitate was collected by filtration and dried over phosphorus pentoxide under reduced pressure to give 0.84 g (84%) of the title compound, mp 194 ° C dec.

The whole procedure was repeated using 10.74 grams (0.0395 mol) of 7-aminocephalosporanic acid, 134 ml of water, 7.5 g of sodium bicarbonate and 7.5 g of 2,3-triazolo-7,0-py12 ridazine-7-amino-2-carboxylic acid. 6-thiol to give 10.53 g (75% yield) of the title compound as crude crystals. This material had a melting point of 189-192 ° C with decomposition.

The same purification was carried out using 10 g of the crude product obtained above, yielding 8.2 g of the purified title compound in a yield of 82%, m.p. 194 DEG C. with decomposition.

Comparative example

7-Amino-3- (pyrido [2,1-c] -s-triazol-3-ylthiomethyl) -3-cephem-4-carboxylic acid

2.72 g (0.01 mol) of 7-aminocephalosporanic acid are suspended in 18 ml of water and a suspension of 2.1 g (0.025 mol) of sodium bicarbonate in 20 ml of water is added dropwise over 10 minutes. The internal temperature of the reaction vessel was then raised to 50 ° C and a suspension of 1.81 g (0.0125 mol) of 3-mercaptopyrido [2,1-c] -s-triazole in 35 ml of acetone was added dropwise over 10 minutes.

The resulting mixture was refluxed at a constant internal temperature of 63 ° C for 4 hours and then cooled with ice. When the internal temperature of the mixture reached 4 ° C, the pH was adjusted to 3.6 by addition of 6 N hydrochloric acid. After stirring for one hour, the precipitated crystals were collected by filtration and dried under reduced pressure with phosphorus pentoxide to give 2.31 g (61%) of the title compound, m.p. 173-175 ° C with decomposition.

Comparative Example 4

Process for the preparation of 7- (D-α-amino-α-p-hydroxyphenylacetamido) -3- (tetrazolo (4,5-b) pyridazin-6-ylthiomethyl) -3-cephem-4-carboxylic acid

'·

NHCOOCH.

3.31 g (0.01 mol) of D - (-) - N-p-methoxybenzyloxycarbonylamino-p-hydroxyphenylglycine are suspended in 25 ml of anhydrous acetonitrile and 1.01 g of N-methylmorpholine are added. The resulting mixture was then cooled to an internal temperature of -9 ° C and a solution of 1.36 g of chlorisobutyl carbonate in 2-3 ml of acetonitrile was added dropwise. Then the resulting mixture was stirred at -5 to -9 ° C for 1.5 hours. Separately, a suspension of 4.01 g (0.011 mol) of 7-amino-3- (tetrazolo [4,5-b] pyridazin-6-ylthiomethyl) -3-cephem-5-carboxylic acid in 20 ml of anhydrous acetonitrile, 6.4 ml of N, O-bis (trimethylsilyl) acetamide are added, followed by stirring at room temperature for 20 minutes to obtain a solution containing the bissilyl compound. This solution was then added all at once to the mixed anhydride solution described above and the mixture was allowed to react for 5 hours at a temperature of -10 ° C to -2 ° C. The mixture is stirred for a further 20 minutes at 0 to 8 ° C and then 20 ml of methanol are added. The insoluble material was collected by filtration, and the filtrate was concentrated and dehydrated to give an oily liquid, to which 1.68 g of sodium bicarbonate and 20 ml of water were added, followed by addition of 20 ml of methanol and 40 to 50 ml of ethyl acetate. homogenous solution. The resulting solution is then concentrated under reduced pressure, mainly to remove methanol, and the solution is then separated into two liquid phases. The resulting aqueous phase was washed twice with 50 ml of ethyl acetate, then 50 ml of fresh ethyl acetate was overlaid. The resulting solution was adjusted to a pH of 2 to 1 by adding 6 N hydrochloric acid with stirring and ice cooling. A rubbery material appears in the solution. The aqueous solution was extracted twice with ethyl acetate, and the resulting ethyl acetate layer was washed with a saturated aqueous sodium chloride solution and dried with magnesium sulfate to give 2.55 g of caramel brown crystals. These crystals contained a small amount of D-N-p-methoxybenzyloxycarbonylamino-p-hydroxyphenylglycine, purified with ethyl acetate and diethyl ether to give 1.38 g of the title compound. The gummy material was mixed with normal butanol to crystallize. 2.6 g of pure title compound are obtained. Prior purification gave 1.38 g of the title compound, yielding a total of 3.98 g of product in 58.7% of theory.

To a solution of 16 ml of trifluoroacetic acid and 4 ml of ice-cooled anisole was added

2.4 g of the title compound described above and the resulting mixture was stirred at room temperature for 35 minutes. To the resulting solution is then added dropwise 350 ml of diethyl ether. The resulting precipitate was filtered, washed with diethyl ether and dried over phosphorus pentoxide under reduced pressure to give 1.94 g (87.5%) of the title compound.

Claims (1)

SUBJECT A process for the preparation of the novel 7- [N-acylamino-aaryl- or thlenylacetamido] -3-heterocyclo-ynyl (methylmethyl) -3-methyl-4-carboxylic acids of the formula I HO-A-CONH-CH-CONH ^R0 (I) cooh where A represents a monocyclic or polycyclic heteroaromatic radical containing at least one nitrogen atom as a heteroatom selected from the group consisting of naphthyridine, pyridopyrimidine, pyridine, pyrimidine or pyridazine, these radicals being optionally substituted by 1 substituent of C1 -C4 alkyl , or (C 1 -C 4) alkylthio, R represents a phenyl radical optionally substituted by 1 or 2 substituents selected from hydroxyl, amino or thienyl, and Het means a tetrazolopyridazine residue, a triazolopyridazine residue or a triazolopyridine residue, as well as non-toxic pharmaceutically acceptable salts of these compounds, characterized in that the compound of formula II is reacted HO — A — COOH {II], where A is as defined above, or a reactive derivative thereof with a compound of formula III H, N-CH-CONH -r ⁱ | R O (lfi) CU ^ S-Hat COOH where R and Het are as hereinbefore defined and the compound is optionally converted to its non-toxic, pharmaceutically acceptable salt.