CS195287B2 - Method of preparing cephalosporin compounds - Google Patents

Description

(54) A method for preparing cephalosporin compounds

The present invention relates to a process for preparing ceralosporin compounds containing an alkoxycarbonyl group at the 3-position of the dihydrothlazine ring of the cephem cyclic system.

The invention relates to a process for the preparation of compounds of formula I, phenyl or benzyl, or X is an azido group or a group of the formula

wherein Z is a group of formula

where

R 5 is hydrogen or C 1-6 alkyl and

Q is hydrogen or alkyl of 1 to 6 carbon atoms, wherein

R is hydrogen, benzhydryl or alkyl of 1 to 6 carbon atoms wherein

R 1 is hydrogen or acyl of the formula

About where

R 1 is an arylalkyl of the formula wherein X is a group of the formula -OR 5, wherein R 3 is an alkyl of 1 to 6 carbon atoms, diphenylmethyl, or X is a group of the formula -SR 4, wherein R 4 is

195

R '(Y) _m -CH ₂ anywhere

R 1 is phenyl or substituted phenyl, wherein the substituents are 1 to 3 halogen atoms, hydroxyl, n-trio or alkyl of 1 to 4 carbon atoms.

Y is O or S and m is 0 or 1, or

R * represents a substituted arylalkyl of the formula H

E kde

R '' is R 'as defined above, 2-thlenyl or 3-thienyl, E is hydroxyl or hydroxyl protected by C 1 -C 4 alkanoyl, carboxyl or carboxyl protected by C 1 -C 6 alkyl, amino or alkoxycarbonyl protected amino (C 1 -C 6) alkoxy; or

R ‘denotes heteroarylmethyl of the formula R" CH 2 "where

R '' is 2-thienyl, 3-thlenyl, 2-furyl or

3-furyl, and when R is a hydrogen atom, a pharmaceutically acceptable nontoxic acid salt, characterized in that the compound of formula II,

Y “T ϊ (Μ

COOR / f where

Rio is Ri as defined above,

R 11 is a carboxyl protecting group and / or

Z ‘is a group of the general formula

O · where

A is bromine, chlorine or a radical of the formula mixed ¹ anhydrydu

O

II — O — C — O — alkyl wherein 1-8 alkyl is C1-C6 alkyl, reacted with a compound of formula

X — T where

X has the characteristics given for formula I above and

T is hydrogen or an alkali metal, and optionally reacting the corresponding 2-cephem derivatives with percarboxylic acid to give the 3-cephem derivatives and the resulting sulfoxide is reduced with phosphorus pentachloride and optionally the 7-acyl group is cleaved by reaction with phosphorus pentachloride and alcohol and optionally re-acylated in the 7-position by reaction with a halide or active ester of an acid R'-CO 2 H, wherein R 'is as defined above, optionally de-esterifying the 3-carboxylic acid and / or 4-carboxylic acid ester in an acid medium and recovering the product optionally in salt 4 -carbonyloxy acids, benzhydryl or C1 -C6 alkyl esters of 4-carboxylic acid.

A variety of cef alosporin series antibiotics are known. These antibiotics all have the same basic cyclic structure, containing a four-membered [beta] -lactam ring attached to a six-membered dihydrothiazine ring, and differ in structure and biological effects from one another. Structurally known eephalosporin antibiotics differ. in the 7-acylamido substituent type and in the 3-position of the dihydrothiazine ring. Desacetoxycephalosporanic acid, for example cephalexin, has a 3-methyl substituent. Many known cephalosporins have a substituted methyl group at the 3-position. The desacetoxycephalosporins have a 3-hydroxymethyl substituent. 3-Alkylthiomethyl a ..

3-heteroarylthiomethylcephalosporins are also described. Recently, certain 3-methoxymethylcephalosporins have been described in U.S. Patent No. 3,665,003 and 3-bromomethylcephalosporins have been described in U.S. Patent Nos. 3,647,788, 3,668,203 and 3,637,678.

In addition to 3-methyl- or 3-substituted methylcephalosporins, 3-formyl-substituted cephalosporins have also been reported. U.S. Pat. No. 3,351,596 of 7. XI. 1976 discloses a Chamberlin process involving the reaction of 3-hydroxymethyl-7-acylamino-3-cephem-4-carboxylic acid with a diazo compound to form an ester

3-formylcephalosporin, after which the ester is reacted with an oxidizing agent selected from the group consisting of manganese dioxide and chromium oxide. A similar reaction is described in Belgian Patent No. 768,653. The corresponding 3-formylcephalosporinsulfoxides are described in US Patent No. 3,674,784.

Another group of cephalosporins differing in the substituent type at the 3-position is described in Dutch Published Application No. 7206,931, which discloses certain 3-unsubstituted cephalosporins, as well as a process for their preparation by decarbonation of the corresponding 3-formyl compounds. /.

The 7-acylamino-3-carboxycephene compounds and their derivatives are prepared by oxidation of acyclic or cyclic acetate esters. 7-acylamino-3-formyl cephem-4-carboxylic acid with N-bromosuccinimide to give the intermediate 7-acylanino-3- [alkoxycarbonyl or 2-bromalkoxycarbonyl] derivative. The latter is then de-esterified to give the corresponding 3-carboxylic acid. The 3-carboxylic acid derivatives, such as esters, thioesters, amides, acyl azides and the like, are prepared by reacting the acid chloride or mixed cephem-3-carboxylic acid anhydride with the appropriate nucleophilic reagent. Removal of the ester protecting group at the 4-position yields new antibiotic active compounds which can be used for. combating Gram-positive and Gram-negative microorganism infections. "

The 7-methoxy compounds are prepared from the 7-methoxy-starting compounds, i.e. the corresponding 7-methoxy-3-carboxylic acid halides or anhydrides.

The terms used in the preceding definition of compounds have the following meanings:

- The term "alkyl of 1 to 6 carbon atoms" includes methyl, ethyl, n-propyl, n-butyl, isobutyl, pentyl, n-hexyl, cyclohexyl and similar aliphatic hydrocarbon chains,

Groups such as hydroxyl, methoxy, ethoxy, mpropoxyl, isopropoxyl, n-hutoxyl and the like are included in the definition of the group —OR 1.

Examples of groups of formula i

how. as defined above, are amino, methylamino, ethylamino, dimethylamino, diethylamino, isopropylamino, butylamino.

when R 'is substituted phenyl in the above definition, R' may be mono- or disubstituted halophenyl, such as 4-chlorophenyl, 2,6-dichlorophenyl, 2,5-dichlorophenyl, 3,4-dichlorophenyl, 3-chlorophenyl, -bromophenyl, 4-bromophenyl, 3,4-dibromophenyl, 3-chloro-4-fluorophenyl, 2-fluorophenyl and the like, mono- or dihydroxyphenyl such as 4-hydroxyphenyl, 3-hydroxyphenyl, 2, 4-dihydroxyphenyl and the like;

4-nitrophenyl, mono- or disubstituted alkylphenyl such as 4-methylphenyl, 2,4-dimethylphenyl, 2-methylphenyl, 4-isopropylphenyl, 4-ethylphenyl, 3-n-propylphenyl and the like.

Also, R 1 is disubstituted phenyl, where the substituents may be different, for example 3-methyl-4-hydroxyphenyl, 3-chloro-4-hydroxyphenyl, 2-methoxy-4-bromophenyl, 4-ethyl-2-hydroxyphenyl, 3-hydroxy- 4-nitrophenyl, 2-hydroxy-4-chlorophenyl and disubstituted phenyl with various substituents.

The term "protected carboxy" refers to a carboxy group protected by one of the commonly used ester-protecting groups for carboxyl groups that block or protect the carboxyl group during the course of the reaction at other functional sites of the compounds. Such protected carboxyl groups are used for easy cleavage by hydrolysis or hydrogenolysis to the corresponding carboxylic acid. Examples of carboxylic acid protecting groups are tert-butyl, benzyl, p-methoxy, alkanoyloxymethyl of 2 to 6 carbon atoms in alkanoyl, β-iodoethyl, p-nitrobenzyl, diphenylmethyl (benzhydryl), phenacyl, p-halophenacyl, 2,2 , 2-trichloroethyl and the like groups forming the ester portion of the molecule. The type of these ester-forming groups is not critical as long as the ester formed is stable under the reaction conditions described below. Preferred carboxyl protecting groups are benzhydryl, p-nitrobenzyl, tert-butyl, [beta], [beta], [beta] -trichloroethyl, p-methoxybenzyl, and [beta] 3-iodoethyl, most preferably benzhydryl, p-methoxybenzyl and tert-butyl.

In the foregoing definitions, the protecting groups for the carboxy group are not exhaustively defined. The function of these groups is to protect the reactive functional groups during the preparation of the desired products and then cleave without disturbing the remaining molecule. Many of these protecting groups are well known in the literature and the use of other groups, such as those described in J.F.W. McOmnie, "Protective." Groups in Organic Chemistry ”Plenum Press, 1973, is also suitable.

Examples of acyl groups as defined above are acetyl, propioyl, butyryl, hexanoyl, heptanoyl, 2-penenoyl, acryloyl, 5-aminoadlpoyl, chloroacetyl, bromoacetyl and the like;

Examples of acyl groups. · O ^; 'II. ·

R 4 -C 8 are benzoyl, 2,6-dimethoxybenzoyl, 4-chlorobenzoyl, 4-methylbenzoyl, 3,4-dichlorobenzoyl, 3-bromobenzoyl, 4-nitrobenzoyl and the like.

Examples of acyl groups

About where

R 1 is a group of the formula R 1 - (Y) m - CH 2 - and m is 0, are cyclohexa-1,4-dien-1-acetyl, phenylacetyl, 4-chlorophenylacetyl, 3-hydroxyphenylacetyl, 3-cyanophenylacetyl, 4-hydroixyl- 3-methylphenylacetyl, 4-bromophenylacetyl, 4188287

ethoxyphenylacetyl, 4-nitrophenylacetyl, 3,4-dimethoxyphenylacetyl and the like; Are acyl groups phenoxyacetyl, 3-hydroxyphenoxyacetyl, 4-chlorophenoxyacetyl, 3,4-dichlorophenoxyacetyl, 2-chlorophenoxyacetyl, 3,4-dimethylphenoxyacetyl, 4-isopropylphenoxyacetyl, 3-nitrophenoxyacetyl and the like substituted phenoxyacetyl groups; are examples of phenylthioacetyl phenylthioacetyl, 2,5-dichlorophenylthioacetyl, 3-chloro-4-fluorophenylthioacetyl, 3-bromophenylthioacetyl and the like acyl groups.

Examples of acyl groups wherein R 1 is substituted arylalkyl of the formula

H

E:

are hydroxy-substituted arylalkyl groups such as 2-hydroxy-2-phenylacetyl of the formula

or a 2-formyloxy-2-phenylacetyl group of the formula

OCHO and the like, wherein the phenyl ring is substituted, such as. for example 2-hydroxy-2- (3-chloro-4-hydroxyphenyl) acetyl, 2-formyl-oxy (2- (4-hydroxyphenyl) acetyl), 2-hydroxy-2- (3-bromophenyl) acetyl, 2-formyl- 2- (3,5-dichloro-4-hydroxyphenyl) acetyl, 2-formyloxy-2- (3-chloro-4-methoxyphenyl) acetyl, 2-hydroxy-2- (3-chlorophenyl) acetyl and the like.

Examples of the acyl group wherein R 'is a carboxyl or alkoxycarbonyl substituted arylalkyl group are 2-phenyl-2-carboxyacetyl, 2-phenyl-2- tert -butoxycarbonylacetyl, 2- (4-chlorophenyl) -2-benzyloxycarbonylacetyl, 2- (3) -nitrophenyl-2-carboxyacetyl and the like

When R * is an amino-substituted arylalkyl or derivative thereof, the acyl groups are 2-amino-2-phenylacetyl, 2-amino-2-phenyl-2-tert-butoxycarbonylamino) acetyl, 2- (4-hydroxyphenyl) -2-aminoacety 1 and similar acyl groups.

Examples of acyl groups O where

R 1 is a heteroarylmethyl group of the formula R 1 -CH 2 -, are 2-thlenylacetyl, 3-thienylacetyl, 2-furylacetyl.

The novel compounds of formula I are cephemes having first order derivatives such as amides, thioesters, esters and acyl azides of the carboxyl group at the G-3 position; prepared by reacting the appropriate nucleophilic ¹ reagent with a mixed 7-acylamido-3-carboxy-3- (or 2) -cephem-4-carboxylic acid ester anhydride or halide. These processes for the preparation of carboxylic acid derivatives are well known in the chemistry of cephalosporin and are common in other fields of organic chemistry. Thus, esters, thioesters and amides are conveniently prepared by reacting the corresponding alcohol, thiol or amine with an active carboxyl functional group, for example an acid halide or mixed anhydride.

Acid halides or mixed anhydrides are prepared from 7-acylamino-3-carboxy-3- (or 2) -phenyl-4-carboxylic acid esters by conventional methods. Generally, the mixed anbydrides are prepared by a two-step process comprising

a) reacting 3-carboxycephem with 1 equivalent of a tertiary amine such as. is triethylamine, N-methylmorpholine or diethylaniline, in an inert organic solvent at a temperature of about -10 ° C to 0 ° C; and

b) reacting the resulting tertiary amine salt with C 1 -C 5 alkyl chloroformates such as methyl chloroformate, ethyl chloroformate, propyl chloroformate, isobutyl chloroformate and the like lower alkyl chloroformates.

Examples of mixed anhydrides obtainable by this process are:

benzhydryl-7- (2-thienylation tamldo) -3-ethylcarbonyldioxycarbonyl-3-cephem-4-carboxylate

4'-Methoxybenzyl 7- (3-thienylation tamldo) -3-methylcarbonyldioxycarbonyl-3-ee-4-carboxylate, tert-butyl-7-phenyiacetamide-3-isobutylcarbonyldioxycarbonyl-3-cephem-4-carboxylate, benzhydryl-7 -phenoxyacetamido-3-ethylcarbonyldioxycarbonyl-3-cephem-4-carboxylic acid ocylate;

4'-Methoxybenzyl-7-acetamido-3-methylcarbonyldioxycarbonyl-3-cephem-4-carboxylate, benzhydryl 1-7- (2-thiazolylacetamido) -3-m-butylcarbonyldioxycarbonyl-3-Ce-4-carboxylate, tert-butyl-7 3-Ethylcarbonyldloxycarbonyl-3-cephem-4-carboxylate and the corresponding 2-cephem compound. These compounds can be reacted in situ with the desired nucleophilic reagent.

The acid halides of the 7-acylamino-3-carboxy-3- (or 2) cephem-4-carboxylic acid esters are prepared by reacting the corresponding halogenating agent and the cephemic acid sodium salt. The sodium salt of the carboxyl group at the C-3 position is simply prepared by suspending a solution of the acid in ethyl acetate with an aqueous solution of sodium bicarbonate. The sodium salt, which is soluble in ethyl acetate, remains in the organic phase, which can be separated, dried with conventional drying agents such as anhydrous sodium sulfate, anhydrous magnesium sulfate or molecular sieves. Thus, the sodium salt can be isolated or reacted in an ethyl acetate solution with the desired halogenating reagent to give the acid halide. Preferably, the sodium salt is isolated and dried prior to use in the preparation of the acid halide. Generally, acid chlorides are prepared by reacting the sodium salt of 7-acylamino-3-carboxy-3- (or 2J-cephem-4-carboxylic acid ester with 2-3 equivalents of oxalyl chloride in methylene chloride in the presence of a few drops of dimethylformamide at 0-5 ° C). C. The acid chloride is then isolated by evaporation of the reaction mixture at low temperature to dryness and used without further purification for the preparation of 7-acylamino-3-carboxy-3- (or 2'-cephem-4-carboxylic acid ester) derivatives. prepared in a similar manner using phosphorous tribromide or thionyl bromide as the halogenating agent Examples of acid halides that can be prepared as described above are:

benzhydryl-7- (2-thienylacetamido) -3-chlorocarboinyl-3-cephem-4-carboxylate, benzhydryl-7- (4-tert-butoxycarbonyl-4-tert-butoxycarbamylbutyl) -3-chlorocarbonyl-3-cephem-4- carboxylate, tert-butyl 7- (4-methoxyphenylacetamido) -3-chlorocarbonyl-3-cephem-4-carboxylate,

4'-methoxybenzyl-7-iodoacetamido-3-chlorocarbonyl-3-cephem-4-carboxylate, benzyl dryl-7- (4-chlorophenylthioacetamido) -3-chlorocarbonyl-3-cephem-4-carboxylate, tert-butyl-7 acetamido-3-chlorocarbonyl-3-cephem-4-carboxylate, benzhydryl 7- (3-thienylacetamido) -3-chlorocarbonyl-3-cephem-4-carboxylate,

4‘-Methoxybenzyl-7-benzamido-3-chlorocarbonyl-3-eephem-4-carboxylate

XO corresponding to the 2-cephem of the compound and the corresponding 2-cephem and 3-cephem-3-bromocarbonyl compounds.

The preparation of the starting compounds, 7-acylamino-3-carboxy-3-cephem-4-carboxylic acid esters 8e can be carried out as follows:

1) preparing 7-acylamino-3-formylcephem-4-carboxylic acid ester;

2) preparing acetal (cyclic or acyclic) of the 3-formyl functionality;

3) converting the acetal to the corresponding cephem-3,4-dicarboxylic acid diester by reacting acephal with N-bromosuccinimide in the presence of a free radical initiator in an inert organic solvent at a temperature of from 20 to 100 ° C;

4) de-esterifying the 3-carboxylic acid ester to give the corresponding 4-alkoxycarbonylcephem-3-carboxylic acid.

The process for preparing 3-carboxy-cephem starting materials can be carried out on materials with side chains most suitable for the preparative process (due to availability or stability to reaction conditions), after which the side chains can be replaced with other 7-acylamine side chains preferred for maximum biological activity by conventional cleavage and reaicylation. The intermediates of these transacylation processes are the corresponding 7-aminocephem derivatives. .

For simplicity, acid halides (chlorides and bromides) and mixed anhydrides of 7-acylamino-3-carboxy-3- (or 2) cephem-4-carboxylic acid esters are referred to below as "activated 3-carboxycephems" or "activated 3-carboxycephemic" starting compounds'.

As mentioned above, acid halides and mixed anhydrides of esters of 7-acylamino-3-carboxy-3- (or 2) cep-4-carboxylic acids are preferred intermediates for the preparation of other cephem compounds which are first-order derivatives of the carboxyl group in the position C-3 of the cephem ring. Thus activated 3-carboxycephemic intermediates are reacted with alcohols to form cephem-3,4-dicarboxylic acid diesters, with thiols to form cephemes with thioester. a group in the C-3 position or with amines or derivatives of amines to form cephemes having an aminocarbonyl (carbamoyl) group or derivatives thereof in the C-3 position. Similarly, reacting the activated 3-carboxycephem with an azide ion (e.g., sodium azide) provides the corresponding acylazide.

Cephem diesters are prepared by reacting alcohols with activated 3-carboxycephemic starting compounds (acid halides or mixed anhydrides) in an inert organic solvent, for example methylchloride, chloroform or tetrahydrofuran, usually in the presence of a basic reagent such as tertiary amine or sodium bicarbonate. For example, benzhydryl-7-phenylacetamido-3-chlorocarbonyl19S '' ¹ '·'

The 2-cephem-4-carboxylate is reacted with

4-methoxybenzyl alcohol with tetrahydrofuran in the presence of excess sodium bicarbonate to give benzhydryl-7-phenylacetamido-3- (4-methoxybenzoxycarbonyl) -2-cephem-4-carboxylate.

Cephemes having a thioester function in the C-3 position can be prepared by reacting the activated 3-carbotxycephem with mercaptan in an inert organic solvent, for example methylene chloride, chloroform, tetrahydrofuran, dioxane or acetonitrile.

This is a standard procedure for the preparation of thioesters, which is well documented in the chemical literature. While the presence of basic reagents such as triethylamine or sodium carbonate is not critical to the preparation of these thloesters, these reagents are used to reduce the acidity of the reaction mixture and thus shorten the reaction time and increase the product yield.

Examples of mercaptans that can be used for the preparation of thloesters are methanthiol, ethanethiol, 2-propanethiol, 1-butanethiol, 2-methyl-2-butanethiol, 1-pentanethiol, 1-hexanethiol, thiophenol, benzylthiol and heteroarylthiols such as 5- tetrazolthiol or 1,3,4-thiadiazole-2-thiol. For example, tert-butyl 7- (2,5-dichlorophenylthio) acetamido-3-ethylcarbonyldioxycarbonyl-3-cephem-4-carboxylate. is reacted with 1.1 equivalents of 2-propanethiol in tetrahydrofuran in the presence of 1 to 10 equivalents of sodium bicarbonate at room temperature to give tert-butyl-7- (2,5-dichlorophenylthio) acetamido-3- (2-propylthio) carbonyl -3-Cephem-4-carboxylate.

Specific examples of thioester intermediates obtainable by the above method are:

benzhy dryl-7-phenoxyacetamido-3- (methylthio) carbonyl-3-cephem-4-carboixylate,

4'-Methoxybenzyl 7-acetamido-3 - [(1-pentylthio) carbonyl] -2-cephem-4-carboxylate, tert-butyl 7- (1,4-cyclohexanedienylacetamido) -3 - [(2-propylthio) carbonyl] 3-cephem-4-carboxylate, benzhydryl-7- (2-thienylacetamido) -3- (phenylthio) carbonyl-3-cephem-4-carboxylate, benzhy dryl-7-phenylacetamido-3- (benzylthio) carbonyl -2-cephem-4-carboxylate, benzhydryl-7 - [(4-chlorophenylthio) acetamido] -3- (1,2,3,4-tetrazole-5-mercapto) carbonyl-3-cephem-4-carboxylate ,

4‘-Methoxybenzyl-7- (2-butenamido) -3 - ((1,

3,4-thiadiazole-2-mercapto) carbonyl] -3-cephem-4-carboxylate and tert-butyl 7- (2-thienylacetamido) -3- (ethylthio) carboinyl-2-cephem-4-carboixylate.

187

Biologically active cephem thioesters are prepared from the above intermediates by conversion to S-cephem derivatives (from the 2-cephem compounds) and cleavage of the C-4 carboxyl protecting group as described above. Examples of active thioesters available as described above are: ...

7- (2-thienylacetamido) -3- (butylthio) carbonyl-3-cephem-4-carboxylic acid,

7-Acetamido-3- (methylthio) carbonyl-3-cephem-4-carboxylic acid

7-iodo-tamido-3- (phenylthio) -acetamido-. -3-cephem-4-carboixylic acid,

7-phenylacetamido-3- (1,3,4-thiadiazole-2-mercaptocarbonyl) -3-cephem-4-carboxylic acid,

7- (1,4-cyclohexyldienylacetamido) -3 - ((2-propylthiO) carbonyl] -3-cephem-4-carboxylic acid,

7- (2-thienylacetamido) -3- (hexylthio-) carbonyl-3-cephem-4-carboxylic acid; and

7- (2-furylacetamido) -3- (ethylthio) carbonyl-3-cephem-4-carboxylic acid.

Activated 3-carboxycephemerivatives are converted to 3-aminocarbonyl- or 3-substituted aminocarbonylcephenes by reaction with 1 to 5 equivalents of an amino group reagent. This technique is a standard procedure for the preparation of amide derivatives and has been applied in many fields of organic chemistry, including the field of antibiotic penicillin and cephalosporin, for example, in the preparation of the amide side chain of these compounds. Because of the susceptibility of the (3-lactam function of the activated 3-carboxycephem derivative to the attack of amines and amino derivatives), the preparation of the 3-amino (unsubstituted amino +) carbonyl compound is preferably carried out at temperatures lower than those at amide formation; The reaction of amines or amino derivatives with activated 3-carboxycephemerivatives (acid chlorides or mixed anhydrides) is preferably carried out at a temperature of -70 ° C to -80 ° C in an inert atmosphere. an organic solvent such as tetrahydrofuran, methylene chloride, chloroform, acetonitrile and the like inert organic solvents.

Certain types of prepared cephem derivative are, of course, based on the type of amine or derivative used in the preparation.

Treatment of the activated 3-carboxycephem starting compound as defined above with ammonia affords the corresponding primary amide, (3-aminocarbonylcephene). These primary amide derivatives can also be prepared by reduction

1952-7 of the corresponding 3-azidocarbonylcephem compounds as described below.

The primary amines are reacted with 3-carboxycephemic acid chloride or the corresponding mixed anhydride under the conditions described above to give the corresponding secondary amide (3-monoalkyl or monoarylaminocarboxylic biphenyls). The primary amines useful for the preparation of these compounds are methylamine, ethylamine, 2-propylamine , butylamine, cyclohexylamine, aniline, p-toluidine, benzylamine and the like primary amines. When secondary amines are used in a similar procedure, the resulting products are tertiary amides (3-disubstituted aminocarbonylcephems). Compounds which may be included within the scope of the term "secondary amine" and which are useful include dialkylamines such as dimethylamine, diethylamine, diisopropylamine, cyclohexylmethylamine, benzylethylamine and di-n-butylamine, arylalkylamines such as N-methylaniline, N ethylaniline, N-methyl-o-toluidine and N-isopropylaniline, cyclic amines such as morpholine, piperidine and pyrrolidine and the like amines.

. Benzhydryl-7-phenylacetamido-3 * ethylcarbonyldioxycarbonyl-3-cephem-4-carboxylate was treated with 2 equivalents of tert-butylamine in tetrahydrofuran at -73 ° C (dry ice bath in acetone) to give benzhydryl-7-phenylacetamido-3- (terc.butylaminokarbosnylj-3-cephem-4-carboxylate. Similarly, morpholine was reacted under the same conditions of 4'-methoxybenzyl 7- (2-thienylacetamido) -3-bromkarbonyl-2-cephem-4-carboxylate ^and carboxylate to afford 4 ' -methoxybenzyl-7- (2-thienylacetamido) -3-morpholinecartionyl-3-cephem-4-carboxylate If tert-butyl 7- (1,4-cyclohexadienylacetamido) -3- (tert-butylcarbonyldioxycarbonyl) -3-cefem-4 The carboxylate is reacted with 2.5 equivalents of N-methylaniline in methylene chloride at -76 ° C for one hour, the product isolated and tert-butyl 7- (1,4-cylcohexadienylacetamido) -3- (n) is isolated by chromatography. (methylanilinocarbonyl) -3-cephem-4'-carboxylate.

Examples of 3-aminocarbonyl compounds available as described above are:

benzhydryl 1-7 - ((2,5-dlchlorophenylthio) acetamido] -3-aminocarbonyl-3-cephem-4-carboxylate, benzhydryl-7-propionamido-3-methylaminocarbonyl-3-cephem-4-carboxylate, tert-butyl- 7- (4-methoxyphenylacetamido) -3- (diisopropylamino) carbonyl-2-cephem-4-carboxylate,

4'-Methoxybenzyl 7- (3,4-dichlorobenzamido) -3-anilnocarbonyl-3-cephem-4-carboxylate, tert-butyl N - (1-tetrazolylacetamido) -3-piperidinecarbonyl-2-cephem-4 -carbhoxylate, benzhydryl 1-7- (2-thienylacetamido) -3-pyrrolidinocarbonyl-3-cephem-4-carboxylate, benzhydryl-7-phenylacetamido-3- (N-methyl-o-toluidinocarbonyl) -2-cephem-4 -carboxylate, tert-butyl-7-chloroacetamido-3-aminocarbonyl-3-cephem-4-carboxylate, and

methoxybenzyl 7 - [(2,5-dichlorophenylthio) acetamido] -3-ethylaminocarbonyl-3cephem-4-carboxylate.

Compounds having an amino group (i.e. compounds with nucleophilic nitrogen atom with active hydrogen atom) other than the amino compounds mentioned hitherto react similarly with the activated 3-carboxycephem compound to give the expected first order derivatives of 3-carboxycephams. Such amino compounds include hydroxylamines, C1-C7 alkoxyamines, hydrazines, phenylhydrazines, C1-C3 hydrazides and quanidines.

In general, the reactions of these compounds are carried out under the same conditions as described above for the reactions of primary or secondary amines with activated 3-carboxycephem derivatives.

Hydroxylamines react with 7-acylamino-3- "activated" carboxy-3- (or 2) cephem-4-carboxylic acid esters at low temperature (-70 ° C to -80 ° C) to give the corresponding hydroxamic acid derivatives. Examples of such hydroxylamine reagents are hydroxylamine, methylhydroxylamine, ethylhydroxylamine, ferylhydroxylamine, propylhydroxylamine and o-tolylhydroxylamine. Alkoxyamines react with activated 3-carboxycephemoyl derivatives to give the corresponding hydroxamic acid esters. Examples of alkoxyamines that can be used in the preparation of hydroxamic acid esters are methoxyamine, ethoxyamine or benzyloxyamine.

Hydroxylamine and its derivatives, as exemplified by the above compounds, are in many cases used or added to the reaction mixture in the form of an acid salt. Common salts used for this purpose are hydrochlorides or p-tosylates (p-toluenesulfonates).

Benzhy dryl-7- (2-thienylacetamido) -3-chlorocarbonyl-2-cephem-4-carboxylate was treated with 1.1 equivalents of methoxyamine in tetrahydrofuran at -76 ° C to give benzhydryl-7- (2-thienylacetamido) -3 -methylaminocarbonyl-4-cephem-a-carboxylate. 4'-Methoxybenzyl-7-butyramido-3-methylcarbonyldioxycarhoriyl-3-cephem-4-carboxylate is reacted with 1.1 equivalents of phenylhydroxylamine in methylene chloride at -76 ° C to give 4'-Methoxybenzyl-7-butyramido-3-phenylhydroxamidocarbonyl-3 -cephem-4-carboxylate.

Examples of hydroxamic acids and their esters are:

13,287 benzhydryl-7- (4-fluorophenylacetamido) -3-hydroxyaminocarbonyl-3-cephem-4-carboxylate,

4'-methoxybenzyl-7- (2-thienylacetamido) -3-methylhydroxaminocarbonyl-2-cephem-4-carboxylate, tert-butyl 7- (4-methoxyphenylacetamido) -3-methoxyaminocarbonyl-3-cephem-4-carboxylate, benzyryl 7 - [(2,5-dichlorophenylthio) acetamido] -3-benzyloxyaminocarbonyl-2-Cefem-4-carboxylate, benzhydryl-7-acetamido-3-ethylhydroxaminocarbonynyl-3-cephem-3-carboxylate, tert-butyl-7- phenoxyacetamido-3- (o-tolylhydroxaminocarbonyl-3-cephem-4-carboxylate),

4‘-Methoxybenzyl-7- (4-cyanobenzamido) -3-ethoxyaminocarbonyl-2-cephem-4-carboxylate

4‘-methoxybenzyl-7- (2-thienylacetamido} -3-phenylhydroxaminocarbonyl-2-cephem-4-carboxylate; benzhydryl-7- (4-trifluoromethylphenylacetamido) -3-methoxyaminocarbonyl-3-cephem-4-carboxylate.

Other nucleophilic nitrogen compounds useful for the preparation of the amide of analogous carboxylic acid derivatives at the C-3 position are hydrazines, phenylhydrazines, hydrazides such as acetylhydrazide or propionohydrazide, and quanidine. These compounds react with the activated 3-carboxycephem starting compound under the same conditions as defined above for the primary and secondary amines, i.e., in an inert organic solvent at low temperature. For example, benzhydryl-7-acetamido-3-ethylcarbonyldioxycarbonyl-2-cephem-4-carboxylate is reacted with 1.0 equivalent of phenylhydrazine in methylene chloride at -76 ° C to give benzhydryl-7-acetamido-3-phenylhydrazinocarbonyl-2-cephem- 4-carboxylate. Similarly, tert-butyl-7-benzamido-3-chlorocarbonyl-3-cephem-4-carboxylate is reacted with 1.0 equivalent of acethydrazide in tetrahydrofuran at -76 ° C to give tert-butyl-7-benzamido-3-acetylhydrazinocarbonyl-3. -cephem-4-carboxylate.

Examples of other cephem compounds obtainable by the reaction of an activated 3-carboxycephemic derivative and guanidine, hydrazine, phenylhydrazine or hydrazide are:

benzhydryl-7- (2-thienylacetamido) -3-guanidinocarbonyl-2-cephem-4-carboxylate, tert-butyl 7-phenoxyacetamido-3-hydrazinocarbonyl-3-cephem-4-carboxylate,

1B

4'-methoxybenzyl-7-chloroeetamido-3-phenylhydrazinocarbonyl-3-cephem-4-carboxylate, tert-butyl 7- (2-thienylacetamido} -3-guanidinocarbonyl-2-cephem-4-carboxylate, :: benzhydryl-7- (2-furylacetamido) -3-hydrazinocarbonyl-3-cephem-4-carboxylate, benzhydryl-7- (2-chlorophenylacetamido) -3-acetylhydrazinocarbonyl-2-cephem-4-carboxylate, and

4‘-Methoxybenzyl 7-propionamido-3-phenylhydrazinocarbonyl-3-cephem-4-carboxylate.

The activated 3'-carboxycephemic starting compounds also react with other nucleophilic agents, such as the azide ion, to form the corresponding acylazide. In general, the reaction with the ionic ion is carried out in a manner similar to the above described reactions of mixed anhydrides and acid halides. The azide ion source is, for example, sodium azide or tetramethylguanidiumazide, which react with the activated 3-carboxylate intermediate in an inert organic solvent such as dioxane, tetrahydrofuran or dimethylformamide at room temperature. The product, 7-acylamino-3-azidocarbonyl-2 (or 3) -cephem-4-carboxylic acid ester, is an important intermediate in the preparation of 7-acylamino-3-aminocarbonyl-3-cephem-4-carboxylic acids, active antimicrobial compounds according to invention. Azidocarbonylcephems are reduced by hydrogenation in methanol at 0.35 MPa in the presence of 5% palladium on carbon (pre-reduced at 15 MPa 15 mintite) to give the corresponding amide, 7-acylamino-3-aminocarboriyl-3- (or 2) cephem ester. -4-carboxylic acids.

Cefem-a-carboxylic acid esters containing first order derivatives on the carboxy group at the 3-position can be converted directly to ¹ biologically active compounds by: a) conversion to the 3-cefem derivative (if the compound is originally 2-cefem); -4 carboxyl group.

The rearrangement of the 2-cephem derivatives to the corresponding 3-cephem compounds is accomplished by an oxo reductive procedure well known in the chemistry of cephalosporin. Generally, this process is carried out by first oxidizing the 2-cephem compound, for example m-chloroperbenzoic acid, to give the corresponding 3-cephem-1-oxide derivative, which is then reduced with a trivalent phosphorus compound such as phosphorus tribromide or phosphorus trichloride, preferably using dimethylformamide solvents.

It will be appreciated that this rearrangement can be carried out in any of several stages of the process for preparing the active compounds. Preferably, the conversion of the 2-cephem compound to the 3-cephem compound can be carried out at a stage where the 2-cephem compound does not have a free carboxyl group or an amino group. Also, for the preparation of particularly active compounds, the conversion of the 2-cephemic acylamino intermediates (with any carboxy or amino-protected optionally present in the acyl group) at the C-7 position with the ester protected with a carbonyloxy group in the 4-position with the ester, thioster or amido group in position C-3. Although the order of rearrangement in the reaction sequences is not critical, then, if conditions permit, it is experimentally most advantageous to carry out the rearrangement until the C-3 substituent corresponds to the final product.

The cleavage of the ester moiety of the molecule at C-4 into the free 4-carboxyl group is carried out in a conventional manner, using specific methods depending on the presence of a particular ester protecting group. For example, benzhydryl, tert-butyl and p-methoxybenzyl groups are readily cleaved by reaction with an acid such as trifluoroacetic acid customary in the presence of a carbonate stabilizer such as anisole. The de-esterification of the N, β, β-trichloroethyl and 2-iodoethyl esters is carried out by reaction with zinc and an acid, such as formic, acetic or hydrochloric acid. Cleavage of the p-nitrobenzyl ester protecting group is usually accomplished. carried out by hydrogenation of the ester in the presence of palladium, rhodium and the like, in suspension or on a carrier such as barium sulfate, coal, aluminum oxide, etc. It should be noted that these methods may also be used to cleave protecting groups which may be present at any location cephem compounds.

The free acids form salts of carboxyl groups with any inorganic and organic bases. Pharmaceutically acceptable salts are formed by reaction of the free acids with bases such as sodium hydroxide, sodium carbonate, potassium hydroxide, potassium 2-ethylhexanoate, calcium carbonate, ethylamine, 2-hydroxyethylamine and the like. Preferred salts are alkali metal salts. The preferred potassium salt base is potassium 2-ethylhexanoate. Salts can be converted to the free acids by acidification. The free acids and their salts are considered equivalent.

Cephem antibiotics are relatively non-toxic compounds that are useful for combating infections in warm-blooded animals when administered parenterally at a pharmaceutically effective non-toxic dose. 3- (substituted) carbonyl-3-cephem compounds can be prepared in liquid pharmaceutical forms, for example, in water, isotonic saline, and the like, and are administered by the intramuscular injection or intravenous route at a dose of 125 mg to 16 g per day depending on body weight patient, disease and other patient-related factors. Infection may be controlled by repeated administration of small doses, in other cases a larger non-toxic dose may be administered. The antibiotic compounds may be administered as ‘free acids or in the form of nontoxic salts such as sodium or dragee salts. A very preferred class of active compounds are compounds of formula:

COOH where

R 1 is an acyl group of formula O and R 3 is hydrogen, alkyl of 1 to 6 carbon atoms:

A particularly preferred group of antibiotics is represented by the above formula wherein R 3 is alkyl of 1 to 6 carbon atoms and R 3 is a group

H

Rio—- C—

KdeΕ where

R 10 is phenyl or 4-hydroxyphenyl and E is hydroxyl, fortoxydioxo, or carboxyl. Examples of such preferred compounds are:

7- (D-mandelamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid,

7- (2-phenyl-2-carboxyacetamido) -3-ethoxycarbonyl-3-cephem-4-carboxylic acid,

7- [D-2- (4-hydroxyphenyl) -2-formyloxyacetamido) -3-n-propoxycarbonyl-3-cephem-4-carboxylic acid,

7- [D-2- (4-hydroxyphenyl) -2-hydroxyacetamido] -3-methoxycarbonyl-3-cephem-4-carboxylic acid,

7- (2-Phenyl-2-carboxyacetamido) -3-cyclopexyloxycarbonyl-3-cephem-4-carboxylic acid,

7- [D- (2-Phenyl-2-formyloxyacetamido)] -3-isopropoxycarbonyl-3-cephem-4-carboxylic acid,

7- (2-phenyl-2-carboxyacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid.

199287

Antimicrobial tests

The compounds of formula I have been tested for activity against certain microorganisms to determine their antimicrobial activity. Tests against the organism identified as X-12, a Gram-positive organism of Bacillus subtilis and Serratia marcescens, a strain called X-99, a Gram-negative organism, were used as tests.

The anti-X-12 assays were performed as a sensitivity disc assay where the compound dissolved, usually at a concentration of 1 mg / ml, and coated onto a 6.35 mm filter paper disc. After the disc is dried, the test is performed by placing the disc on the culture of the test organism on the plate. The test results are given as the mean of the zones in which the growth of the organism is inhibited.

Tests against X-99 were performed by allowing the test organism to grow in nutrient solutions containing low concentrations of the compound. Results are reported as the lowest concentrations that inhibit the growth of the organism.

The test results of typical compounds of formula I are as follows:

X-99 X-99

7- (2-thienylacetamido) -3-cephem-3,4-dicarboxylic acid 34 mm 7- (2-thlenylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid 41 mm - ) ··· ' Ϊ Wfe. methyl 7- (2-thienylacetamido) -3-carboxy-3-cephem-4-carboxylate 21 at 10 mg / ml and 7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid sodium salt > 200 µg / ml 7- (2-phenyl-2-formyloxyacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid sodium salt - 16.5 7- (2-phenyl-2-carboxyacetamido) disodium salt - 3-methoxycarbonyl-3-cephem-4-carboxylic acid acid 33 mm 1 7- [D- (2-phenyl-2-formyl-oxyacetamido)] -3- -methoxycarbonyl-3-cephem-4-carboxylic acid acid 40 and 7- (2-phenyl-2-formyl-oxyacetamido) -3- (4-; (nitrobenzyloxycarbonyl) -3-cephem-4-carboxylic acid 42 7 7- (2-phenyl-2-formyl-oxyacetamido) -3-cephem-3,4-dicarboxylic acid 130 7- (2,5-dichlorophenylthioacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid 41 22nd benzhydryl 7- [D- (2-phenyl-2-formylmethoxyacetamido)] -3-carboxy-3-cephem-4-carboxylate 21 at 5 mg / ml 7- (2-phenyl-2-carboxyacetamido) -3-cephem-3,4-dicarboxylic acid 20 May 7- (2-thienylacetamido) -3- (2-bromoethoxycarbonyl) -3-pefem-4-carboxylic acid 31 7- [D- (2-phenyl-2-formyl-oxyacetamido)] -3- (2-bromoethoxycarbonyl) -3-cephem-4-carboxylic acid 39 3.5 7- (2-phenyl-2-carboxyacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid 80

X-99 X-99

7- [D- (2-phenyl-2-aminoacetamido)] - 3-methoxycarbonyl-3-cephem-4-carboxylic acid 7- (2-phenyl-2-formyloxyacetamido) -3- (4-nitrobenzyloxycarbonyl) sodium salt -3-cephem-4-carboxylic acid 'i'. Λ '

7- (2-thienylacetamido) -3- (N, N-dimethylcarboxamido) -3-cephem-4-carboxylic acid µg / ml mm

Preparation 1

Benzhydryl-7- (2-thienylacetamido) -3-formyl-2-cephem-4-carboxylate

To a suspension of 7- (2-thienylacetamido) -3-hydroxymethyl-2-cephem-4-carboxylic acid (23.6 g, 67 mmol in 500 mL of ethyl acetate) was added dropwise a solution of diphenyldiazomethane (19.4 g, 0.1 mol). in 50 ml of ethyl acetate. the reaction mixture was heated to reflux 15 minutes, cooled to room temperature and evaporated vevakůuk dryness. the residue was -promyje 1 liter of ethyl ether and petroleum ether 1: 1 to afford a pink _solid:; benzhydryl 7- ( ^2-i oi thienylacetamido) -3-hydro xymethyl ^and 2-cephem-4-carboxylate (33 g, 94.2% yield).

To a stirred solution of the benzhydryl ester in 1 liter of acetone was added dropwise 33.6 ml (76 mmol,

1.2 eq) of chromic acid. The reaction mixture was stirred at room temperature for 8 minutes. Isopropanol (35 ml) was then added and the mixture was stirred for an additional 5 minutes. The reaction mixture was concentrated in vacuo to a small volume and extracted with ethyl acetate (2 X 40 mL). The organic extracts were combined and washed successively with water (4X), sodium bicarbonate solution, water, 1N hydrochloric acid and brine, and then dried (Na 2 SO 4). Evaporation in vacuo to dryness yielded 31.3 g (95.4%) of crude benzhydryl-7- (2-thienylacetamido) -3-formyl-2-cephem-4-carboxylate, which was purified by either crystallization from toluene (4S). or silica gel chromatography (50 g) using a benzene-ethyl acetate gradient (22 g, 62% yield). The product was recrystallized from methylene chloride-hexane to give white needles (mp 149-150 C): IR (CHC) 1785 (YJ-lactam C = 0), 1680 (amide C-0) and 2830 ^cm-1 (formyl C = O); NMR (CDCl 3) 3.80 (s, 2, CH 2 sidechain), 5.12 (d, 1, J =) 4.0: Hz, C 6 -H), 5.40 (q, 1, J-4) , 0 and 8.0 Hz, C 7 -H), 5.51 (s, 1, C 4 -H) and 9.20 (s, 1, CHO). Analysis for C27H22N2O5S2:

H, 4.28; N, 5.40. Found: C, 62.33; H, 4.19; N, 5.17.

Preparation 2 ....... _______

Benzhydryl-7- (2-thienylacetamido) -3- (1,3-dioxolan-2-yl) -2-cepine-4-carboxylate

Benzhydryl 7- (2-thienylacetamido) -3-formyl-2-cephem-4-carboxylate (21.5 g, 41.5 mmol) was treated with 11.6 mL of ethylene glycol (0.2 mol) and p-toluenesulfonic acid monohydrate. acid (0.197 g, 1.04 mmol) in 500 mL benzene. The mixture was heated to boiling for 10 hours using a Dean-Stark trap (1.5 mL of water was collected), cooled and evaporated to dryness in vacuo. The product is taken up in ethyl acetate and washed successively with sodium bicarbonate solution (2X), water (2Xj) and brine, then dried over sodium sulfate. Evaporation in vacuo to dryness affords the product which is chromatographed on 40 g silica gel using a benzene gradient and Crystallization of the purified product from methylene chloride and hexane afforded benzhydryl-7- (2-thlenylacetamido) -3- (1,3-dioxolan-2-yl) -2-cephem-4-carboxylate as colorless needles (15, 15). 07 g (64,2θ / ο):

mp 142-143 ° C,. IR (CHCl 3 1780 cm -1 ⁾ (δ-lactam C = 6),

NMR (CDCl3) 3.3-3.9 (m, 4, --CH2 --CH2 -), 3.83 (s, 2, side chain CH2),

5.10 (d, 1, J = 4.0 Hz, Ce-H), 5.17 (s, 1, acetal CH), 5.21 (5.1, Ca-H) and 5.45 ppm ( q, 1, J = 4.0 and 8.0 Hz, C7-H).

For C29H26O6S2: C, 61.69; H, 4.66; N, 4.98. found, C 61.69, H 4.43, N 5.10.

P rin g 3

Benzhydryl-7- (2-thienylacetamido) -3- (2-bromoethoxycarbonyl) -2-cephem-4-carboxylate

Benzhydryl-7- (2-thienylacetamido) -3- (1,3-dioxolan-2-yl) -2-cephem-4-carboxylate (15.07 g, 25.8 mmol) was mixed with N-bromosuccinimide (15.07 g, 25.8 mmol). 5.25 g, 29.5 mmol) and azobisisobutyro24

198287 nitrile (36.5 mg, 0.25 mmol, 0.01 eq) in 1,200 mL benzene. The mixture was heated to gentle boiling for 20 minutes, cooled and evaporated to dryness in vacuo to give a dark colored product. Chromatography on 30 g of silica gel with a toluene / ethyl acetate gradient gives 7.61 g (44.4%) of benzhydryl 7- (2-thienylacetamido) -3- (2-bromomethoxycarbonyl) -2-cephem-4-carboxylate, mp 129-130 ° C, IR (CHCl 3) 1785 cm -1 ( ¹ H -lactam C = O), NMR (CDCl 3)

3.25 (t, 2, J = 6.0 Hz, CHžBr), 3.83 (s, 2, side chain CH2), 4.30 (t, 2, J ^=! 6.0 Hz, O-CH2 -), 4.95 (d, 1, J ^=! 4.0 Hz, C H)

5.45 (q, 1, J = 4.0 and 8.0 Hz, C7-H), 5.50 (s, 1, C4-H) and 7.80 ppm (s, 1, C2-H) . Analysis for C29H28BrN2O6S2:

calculated C 54.29, H 3.93, N 4.37. Found C 54.22, H 3.90, N 4.27.

Preparation 4 '

Benzhydryl-7- (2-thienylacetamido) -3- (2-iodoethoxycarbonyl) -2-cephem-4-carboxylate

Benzhydryl-7- (2-thienylacetamido) -3- (2-bromoethoxycarbonyl) -2-cephem-4-carboxylate (7.61 g, 12 mmol) was treated with sodium iodide (6.75 g, 45 meq) in 100 ml acetone. The reaction mixture was degassed and then heated to 35 ° C with stirring for 16 hours. The reaction mixture was then filtered and evaporated to dryness. The residue was dissolved in ethyl acetate, washed with water (3X), brine and dried over Na 2 SO 4. Evaporation in vacuo to dryness yielded 7.78 g (95.5%) of benzhydryl-7- (2-thienylacetamido) -3- (2-iodoethoxycarbonyl) -2-cephem-4-carboxylate: IR (CHGI3) 1785 cm @ -1. ¹ (δ-lactam C = O), NMR (CDCl 3) 2.96 (tt 2, J = 7.0 Hz, CH 2 I), 3.80 (s, 2, side chain CH 2), 4.24 µ , 2, J = 7.0 Hz, —OCHa—),

4.95 (d, 1, J · = Ί 4.0 Hz, Ce-H), 5.24 (q, 1, J = 4.0 Hz, C7-H), the remainder of the signal overlaid with C4-H), 5.50 (s, 1, C4H), and 7.80 ppm (s, 1, C2-H).

Preparation 5

Benzhydryl-7- (2-thlenylacetamido) -3-carboxy-2-cephem-4-carboxylate

Benzhydryl-7- (2-thienyl-acetamide) -3- (2-iodoethoxycarbonyl) -2-cephem-4-carboxylate (2.79 g, 4.05 mmol) was dissolved in a mixture of 8 mL glacial acetic acid and 48 mL. dimethylformamide at 0 ° C and then treated with 2.79 g of zinc dust (10.5 eq.)

1.5 hours. The reaction mixture was diluted with ethyl acetate and filtered through celite. The filtrate was washed successively with sodium bicarbonate solution (3X), water, 1N hydrochloric acid, brine and then dried over anhydrous sodium sulfate. Evaporation in vacuo to dryness yielded

1.92 g (89%) of benzhydryl-7- (2-thienylacetamido) -3-carboxy-2-cephem-4-carboxylate NMR (CDCl 3) 3.84 (s, CH 2 sidechain),

4.99 (d, 1, J = ^f 4.0 Hz, C 5 -H), 5.45 (m, C 4 -H and C 7 -H) and 7.80 ppm (s, C 2 -H).

Preparation 6

Benzhydryl-7- (2-thienylacetamido) -3-ethylcarbonyldioxycarbonyl-2-cephem-4-carboxylate

To a cooled (1010 ° C), stirred solution of benzhydryl-7- (2-thienylacetamido) -3-carboxy-2-cephem-4-carboxylate (0.267 g, 0.5 mmol) in 20 mL of methylene chloride was added under argon atmosphere. 0.051 g triethylamine (0.5 mmol). After stirring at -10 ° C for several minutes, the mixture was cooled to -20 ° C and 0.152 g (1.5 mmol) of ethyl chloroformate was added. The reaction mixture was stirred at -20 ° C for 30 minutes and then allowed to warm to 0 ° C. Cold ethyl acetate was added and the resulting solution was washed successively with cold water, cold 1N hydrochloric acid, cold sodium chloride solution and then dried over anhydrous sodium sulfate. Evaporation in vacuo to dryness yielded 283 mg (93.5%) of mixed anhydride. in the form. colorless foam. IR (CHCl) 1798 cm @ ^-1 (lactam Ji-C = 0) NMR (CDCl₃) 1.34 (t, 3, J - 7.0 Hz, CH2CH3),

3.80 (s, 1, CH 2 sidechain), 4.30 (q, 2, J = 7.0 Hz, CH 3 CH 3), 5.02 (d, 1, J = 4.0 Hz, Ce-H) ), 5.40 (q, 1, 4.0 and 8.0 Hz, C7-H), 5.55 (s, 1, C4-H), and 4.72 ppm (s, 1, C2-H) ). Example 1

Benzhydryl-7- (2-thienylacstamido) -3-azidocarbonyl-2-methyl-4-carboxylate

To a stirred solution of benzhydryl-7- (2-thienylacetamido) -3-ethylcarbonyldioxycarbonyl-2-cephem-4-carboxylate (0.283 g, 0.468 mmol) in 20 mL of tetrahydrofuran at room temperature was added 0.12 g of sodium azide (1.85). mmol). The mixture was stirred at room temperature for 10 minutes and then transferred to a separatory funnel with dilution with ethyl acetate. The solution was washed with water, brine and then dried over anhydrous sodium sulfate. Evaporation to dryness afforded 265 mg of acylazide as a brown foam. IR (CHCl3)

O «,

2143 (C-Ns) and CMI ¹ 1785 (/ S-lactam C = 0); NMR (CDCl 3) 3.77 (s, 2, CH 2 sidechain), 4.95 (d, 1, J = 4.0 Hz, C 6 -H, 5.35 (q, 1, J 1 = 4.0) and 8.0 Hz, C7-H), 5.49 (s, 1, Ct-H) and 7.72 ppm (s, 1, C2-H).

Example 2

Ben zhy dryl-7- (2-thienylacetamido) -3-carbamoyl-2-cephem-4-carboxylate

A solution of benzhydryl-7- (2-thienylacetamido) -2,3-aza: dicyclohexyl (0.253 g, 0.34 mmol) in 2 mL of methylene chloride and 50 mL of methanol was treated with 0.267 g of 5% palladium on of carbon (pre-reduced for 15 minutes at 50 psi hydrogen) in 30 mL of ethanol. The hydrogenation of the acylazide is carried out at 0.35 MPa at room temperature for 3 hours. The mixture is filtered. The filtrate is evaporated to dryness, the product is dissolved in a mixture of methylene chloride and acetone. The resulting solution was filtered through celite, and the filtrate was evaporated to dryness in vacuo to give 0.179 g (74.5%) of benzenesulfonyl-7 - (2-thiopheneateamido) -3-carbamoyl-2-cerium-4-carboxylate in white solid, identical to the product prepared by reaction of ammonia and the corresponding 3-carboxylic acid chloride.

Example 3

Benzhydryl-7- (2-thienylacstamido) -3- (phenylthio) carbonyl-2-cephem-4-carboxylate

To a cooled (1010 ° C), stirred solution of benzoylryl-γ-γ-thlenylaioetamido) -3-carboxy-2-cephem-4-carboxylate (0.267 g, 0.5 mmol) in 25 mL of methylene chloride was added 0.152 under argon. g (1.5 mmol) of N-methylmorpholine. The reaction mixture was then cooled to -20 a and 0.135 g (1.25 mmol) of ethyl ether formate was added. The mixture was stirred at 0 ° C for 30 min, then the mixture was cooled to -10 a and 0.165 g of thiophenol was added. After stirring the reaction mixture at 0 ° C for 6 hours, ethyl acetate was added and the resulting solution was washed successively with aqueous sodium bicarbonate solution, water, brine and then dried over anhydrous sodium sulfate. Evaporation in vacuo to dryness afforded the product which was chromatographed on silica gel using a toluene / ethyl acetate gradient. This afforded 0.208 g (66.5 percent) of the title product. IR (CHCl3) 1798 cm- ¹ (zMacat C10); NMR (CDCl 3) 3.80 (s, 2, CH 2 sidechain), 5.06 (d, 1, J 1 = 4.0 HZ, C 6 -H), 5.42 (q, 1, J 4, 0 and 8.0 Hz, C 7 -H), 5.70 (s, 1, Gr-H), and 7.87 ppm (s, 1, G 2 -H).

Example 4

Benzhydryl-7- (2-thlenyl-acetamide) -3- (N, N-dimethylcarboxainido) -2-cis-4-carboxylate

To a cooled (5 ° C), stirred solution of benzhydryl-7- (2-thienylacetamido) -s-R-carboxy-4-carboxylate (0.724 g, 1.305 mmol) sodium salt in 40 mL methylene chloride and two drops A solution of oxalyt chloride (0.430 g, 3.4 mmol) in 3 mL of methylene chloride was added to dimethylformamide. The mixture was stirred under cooling for one hour, then evaporated to dryness in vacuo at low temperature. The residue was dissolved in 25 mL of methylene chloride and cooled to -73 °. To this was added a solution of dimethylamine (0.147 g, 3.58 mmol) in 3 mL cold methylene chloride. the dark brown solution was stirred at -73 ° C for 20 minutes, then 3 mL of 1N hydrochloric acid was added. The reaction mixture was allowed to warm to 10 ° C, ethyl acetate was added and the resulting solution was washed successively with 1N hydrochloric acid, sodium bicarbonate solution (2X), sodium chloride solution, and dried over sodium sulfate. Evaporation in vacuo to dryness afforded 0.883 g of a brown solid which was recrystallized from acetone-hexane to give 0.369 g (51%) of benzhydryl-7- (2-thienylacetamido) -3- (N, N-dimethylkatboxamido) - 2-cephem-4-carboxylate as yellow-brown flakes. NMR (DMSO-d 6) 2.70 (δ, 6, N (CH 3) 2),

3.75 (s, 2, side chain C1-h), 5.12 (d, 1, J = 4.0 Hz, Cs-H) and 5.52 ppm (m, 2,

7-H (CnH).

Analysis for C29H27N3O5Š2:

calculated C 62.46, H 4.16, N 7.54; Found C 62.22, H 4.3.37, N 7.46.

Example 5

Benzhydroxy-7- (2-thienylmethylphetainido) -3- (N, N-dimethylcarboxamide) -3-Cefefli-4-carboxylate

Example 4 was oxidized with m-Chloroperbenzoic acid to give the corresponding 3-cephemesulfoxide. The sulfoxide is then reduced with phosphorus trichloride.

The compound 3-cephem ester was isolated in 39.6% yield from the corresponding 2-cephem compound: IR (CHCl) 1788 cm ^-1 () 3-lactam C = 0) NMR (CDC13) 2.66 (s, 6, N (CH3) 2), 3.35, 3.67 (ABq, 2, J = 18.0 Hz, Ca-H), 3.84 (s, 2, CH2 side chain), 4.95 (d, 1) J = 4.0 Hz, Cs-H) and 5.86 ppm (q, 1, J = 4.0 and 8.0 Hz, C7-H).

Example 6 ·

7- (2-thienylacetamido) -3- (N, N-dimethylcarboxamido) -3-methyl-4-carboxylic acid

To a cooled (5 ° C) stirred solution of 0.16 g of benzhydryl-7- (2-thienylacetamido) -3- (N, N-dimethyicarboxamides) -3-cephem-4-carboxylate in 5 ml of anisole is added 5 ml of cold acid. trifluoroacetic. The mixture was stirred for 10 minutes while cooling, and 40 ml of n-heptane was added. Concentration in vacuo to a small volume gave a precipitate which was filtered off and dissolved in acetone. The acetone solution was filtered and the filtrate was evaporated to dryness. The product was dissolved in ethyl acetate and extracted with sodium bicarbonate solution. The aqueous extracts were combined, overlaid with cold ethyl acetate and acidified with 1N hydrochloric acid. The organic phase was separated, washed with brine and dried

19SJ & 87 27 with sodium sulfate. Evaporation in vacuo to dryness gave 24 mg (21 ´%) of acid. Example7

Benzhydryl-7- (2-thienylacelamido) -3-methoxycarbonyl-2-cephem-4-carboxylate

NMR (CDCl 3), 3.67 (s, CO 2 CH 3), 3.84 (s, CH 2 side chain), 5.00 (d, J = 4.0 Hz, C 6 H), 5.00 (m, J = 4.0 and 8.0 Hz, C7-H) and 7.66 ppm (s, C2-H).

Example 8

Benzhydryl-7- (2-thienylacetamido) -3-isopropoxycarbonyl-2-cephem-4-carboxylate

IR (CHCI3J 1790 cm ^-1 (/ S-lactam); NMR (CDCl3) 1.14 (2 overlapping doublets, 6, -CH (CH3) 2), 3.84 (s, 2, side chain CH2) 5.00 (m, 2, C 6 -H and —CH (CH 3) 2), 5.45 (q, 1, J = 4.0 and 8.0 Hz, C 7 -H), 5.55 (m, 1, C4-H), 6.9-7.5 (m, benzhydryl ArH and thienyl) and 7.72 (m, 1, C2-H).

Example 1 a d 9

Benzhydryl-7- (2-thienylacelamido) -3- (n-propoxycarbonyl) -2-cephem-4-carboxylate

IR (CHCl3) 1790 cm @ ^-1 (/ S-lactam C = 0) NMR (CDCI3J 0.71 (m, 3, CH3), 1.30 (m, 2, OCH2CH2CH3), 3.7 l (s, 2 , side chain (CH 2 -), 3.90 (m, 2, OCH 2 CH 2 CH 3), 4.86 (d, 1, J = 4.0 Hz, C 6 -H), 5.35 (q, 1, J> = ! 4.0 Hz and 8.0 Hz, C7-H), 5.44 (d, 1, J = 1.0 Hz, C4-H) and 7.65 ppm (d, 1, J ^-! 1 (Hz, C2-H) Example 10

Methyl 7- (2-thienylacetamido) -3-benzhydryloxycarbonyl-2-cephem-4-carboxylate

NMR 3.58 (s, 3, CO 2 CH 3), 3.78 (s, 2, side chain CH 2), 5.08 (d, 1, J = 4.0 Hz, Ce-H), 5.42 (s 1, C4-H), 5.57 (q, 1, J = 4.0 and 8.0 Hz, C7-H) and 7.75 ppm (s, 1, C2-H),

They did

Benzhydryl-7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylate

A solution of m-chloroperbenzoic acid (2.34 g, 11.5 mmol) in 40 mL of chloroform was added dropwise over 15 minutes to a cooled (ice bath for 10 minutes) stirred solution of benzhydryl-7- (2-thienylacetamido) -3-methoxycarbonyl-2. -cephem-4-carboxylate (5.72 g, 10.25 mmol) in 250 mL of chloroform. After two hours, the reaction mixture was washed with sodium bicarbonate solution (3X), brine, dried over sodium sulfate and evaporated to dryness in vacuo to give the corresponding 3-cephemulfoxide (thin layer chromatography - one spot). The product is dissolved in 100 ml of dimethylformamide (cooled in an ice bath) and then 1.37 ml of phosphorus pentachloride (15.7 µm.pl,

1.5 eq.]. The mixture was removed from the ice bath and then allowed to react at room temperature for 45 minutes. Ethyl acetate was added to the reaction mixture, and the resulting solution was washed sequentially with sodium bicarbonate solution (3X), water (2X) and brine. After drying over anhydrous sulfate. The solution was evaporated to dryness in vacuo. The crude product is chromatographed on 10 g of silica gel using a toluene / ethyl acetate gradient. There was thus obtained 3.82 g (66.8%) of benzhydryl-7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylate as a yellow solid, mp 201 ° C, dec. (methyl lenchlorldhexan), UV max (EtOH) 283 nm (ε 13.6 X10 ³⁾ inflection 243 nm, IR (CHCl3) 1793 cm @ ^-1 (.beta.-lactam C = 0) NMR (DMSO-d 6) 3.30 (s, 3, CO 2 CH 3), 3.74 3.80 (m, 4, side chain CH 2, C 2 -H), 5.25 (d, 1, J = 1 5.0 Hz, Ce-H 5.90 (q, 1, J = 4.0 and 8.0 Hz, C7-H1) and 7.0, 7.4 ppm (m, 13, benzhydryl, ester, thienyl).

Example 1 a d 1 2

7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid

To a cooled (5 ° C) suspension of benzhydryl-7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylate (0.316 g, 0.579 mmol) in 5 mL of anlsol was added 5.0 mL of cold trifluoroacetic acid. The solid dissolved immediately and the resulting colorless solution was stirred under cooling for 10 minutes, then 50 ml of n-heptane was added. The resulting solution was concentrated in vacuo to a small volume and a white solid precipitated. This solid was filtered off and then dissolved in acetone. The acetone solution was evaporated to dryness in vacuo. The residue was dissolved in ethyl acetate and extracted three times with cold sodium bicarbonate solution. The aqueous extracts were combined, overlaid with ethyl acetate and acidified with 1N hydrochloric acid. The organic layer was separated, washed with brine and dried (Na 2 SO 4). Evaporation in vacuo to dryness gave 0.214 g (97%) of 7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid.

Example 13

7- (2-thlenylacetamido) -3- (isopropyloxycarbonyl) -3-cephem-4-carboxylic acid

Benzhydryl-7- (2-thienylacetamido) -3-isopropoxycarbonyl) -3-cephem-4-carboxylate was de-esterified (83%) with the trifluoroacetic acid-anisole mixture described in Example 10 to give the title compound as a form white solids. The product exhibits biological activity against both gram-positive and gram-negative organisms.

Example 1 4

Benzhydryl-7- (2-thienylacetamido) -3- (n-propoxycarbonyl) -3-cephem-4-carboxylate ¹ To a cooled (5 ° G) stirred solution of 0.561 g benzhydryl-7- (2-thienylacetamido) -3- (n-propoxycarbonyl) -2-cephem-4-carboxylate in 50 ml of chloroform was added a solution of 0.218 g of m-chloroperbenzoic acid in 2 ml of chloroform. After 12.5 hours during which the mixture was allowed to warm to room temperature, the reaction mixture was washed with sodium bicarbonate solution (2X) and brine, and then dried over anhydrous sodium sulfate. Evaporation to dryness gave the product which was chromatographed on 5.0 g of silica gel. The purified product thus obtained is dissolved in 25 ml of dimethylformamide. The solution was cooled in an ice bath, and 0.214 ml of phosphorus trichloride was added. The reaction mixture was stirred without cooling for one hour and then transferred into ethyl acetate in a separatory funnel. The mixture was washed successively with sodium bicarbonate solution (3X), water (4X) and brine (2X), and then dried over anhydrous sodium sulfate. Evaporation in vacuo to dryness gave the product which was chromatographed on 5.0 g of silica gel using a gradient of toluene and ethyl acetate. 0.328 g (58%) of benzhydryl-7- (2-thienylacetamido) -3- (m-propoxycarbonyl) -3-cephem-4-carboxylate was obtained as a light yellow solid. IR (CHC) 1800 ^cm-1 (C = 0 0laktam) NMR (CDCbj 0.65 (m, 3, -GH2CH2CH3), 1.18 (m, 2, -OCHžCHzCHs), 3.68 (s, 2, side chain CH 2), 3.68 (m, 4, -OCH 2 CH 2 CH 3 and C 2 -H), 4.85 (d, 1, J = 5.0 Hz, C 6 -) and 5.82 ppm (q, 1, I '= 5.0 and 9.0 Hz, Gz-H).

Example 15

7- (2-thienylacetamido) -3- (m-propoxycarbonyl) -3-cafem-4-carbaxylic acid

Benzhydryl-7- (2-thienylacetamido) -3- (n-propoxycarbonyl) -3-cephem-4-carboxylate was de-esterified (65%) with a mixture of trifluoroacetic acid and anisole according to the procedure of Example 10 to give the title acid in form of white cloth. The product exhibits biological activity against Gram-positive and Gram-negative organisms.

Example 16

Benzhydryl-7-amino-3-methoxycarboinyl-3-cephem-4-carboxylate

The same side chain cleavage procedure is as described in the example

13. The corresponding 2-thienylacetamido compound is used as starting material. The cephem compound diester is isolated in 93% yield (one spot by thin layer chromatography). IR (CHC) 1795 cm @ ^-1 (.beta.-lactam C = 0) NMR (CDCl3) 1.84 (br s, 2, NH2, exchangeable with D2 O)

3.27 (s, 3, CO2CH3), 3.41, 4.90 (ABq, 2, J = 18.0 Hz, C2-H), 4.70 (d, 1, J = 50 Hz) C 6 -H), 4.92 (d, 1, J = 5.0 Hz, C 2 -H), 7.11 (s, 1, benzhydryl CH) and 7.4 ppm (s, 10, benzhydryl ArH) .

Example 17

Benzhydryl-7- [D- (2-phenyl-2-formyloxyacetamido)] -3-methoxycarbonyl-3-cephem-4-carboxylate

Following the same acylation procedure as described in Example 14, only benzhydryl-7-amino-3-methoxycarbonyl-3-cephem-4-carboxylate was used as the starting material. Chromatography yielded 0.211 grams (61.2%) of product which crystallized from methylene chloride and hexane as white needles (ht 207-208 ° C): IR (CHCl3) 1800 ^cm-first (£ lactam C = 0) NMR (acetone de) 3.34 (s,

3, CO2CH 3), 3.7 (m, C 2 -H), 5.15 (d, 1, J = 5.0 Hz, C 6 -H), 5.95 (q, J 1 = 5.0 and 9.0 Hz, Cz-H),

6.30 (s, 1, CH side chain), 7.10 (s, 1, benzhydryl CH), 7.5 (m, 10, benzhydryl ArH), 8.34 (s, 1, CHO), and 8 43 ppm (d, 1, J = 9.0 Hz, NH).

· Ϊάά

Analysis for C31H26N2O8S:

calculated C 63.47, H 4.47, N 4.78, found C 63.40, H 4.73, N 4.53.

Example 18

7- [D- (2-phenyl-2-formyl-acetamido)] -3-methoxycarbonyl-3-cephem-4-carboxylic acid

The same procedure is carried out as in the example

4, wherein the benzhydryl ester is de-esterified by reaction with trifluoroacetic acid in anisole. The product was crystallized from acetone, methylene chloride and hexane to give white crystals (m.p. 177-178 ° C).

Analysis for C18H16N2O8S:

H, 3.84; N, 6.66.

Found C 51.70, H 4.06, N 6.77.

Example 19 ..... _ _____________ __

Benzhydryl-7 - [(2,8-dichlorophenylthio) acetamido] -3-methoxycarbonyl-3-cephem-4-carboxylate

To cooled (5 ° C), stirred solution of benzhydryl-7-amino-3-methoxycarbonyl199287

32

-S-Cephem-a-carboxylate (0.068 g, 0.632 mmol) in 30 mL of tetrahydrofuran was added. 0.239 g (28.4 mmol) of sodium hydrogen carbonate and 0.390 g (1.52 mmol) of 2,5-dichloro-thiophenoic acid chloride. The mixture was stirred with cooling for one hour. Ethyl acetate was added and the resulting solution was added. washed with water and brine and dried over anhydrous sodium sulphate. Chromatography on silica gel using a toluene / ethyl acetate gradient afforded 0.288 g (71%) of the title compound as a white solid, which crystallized from methylene chloride / hexane to give 173 mg of colorless crystals (b.p. 170-180 DEG C.-IC. CHGls) 1805 ^cm-1 NMR (GDCis, acetone-d 6j, 3.30 (s, 3, CO2CH3}, 3.50, 3.85 (AB q, 2, J = '16 Hz, G-H)

I p

AND

3.93 (s, 2, S-CH2-C), 5.15 (d, 1, J = 4.0 Hz, C6-H) and 5.85 ppm (q, 1, J * = -4, 0 and 8.0 Hz, C7-H).

Analysis for ¹ C ⁵⁰ H ¹⁵ F ³ O ⁵ S ² Cl 2;

C, 55.99; H, 3.76; N, 4.35;

Found: C 56.02, H 3.79, N 4.17.

The corresponding 7- ((2,5-dichlorophenylthio) acetamido] -3-methoxycarbonyl-3-cephem-4-carboxylic acid is prepared by cleavage of the benzhydryl ester with trifluoroacetic acid and anisole as described in Example 4. The product shows antimicrobial activity.

Example 20

Benzhydryl-7- (2-phenyl-2-tert-butoxycarbonylacetamide) -3-inoxycarbonyl-3-methyl-4-carboxylate

Following the procedure of Example 14, acylation of benzhydryl-7-amino-3-methoxycarbonyl-3-cephem-4-carboxylate is acylated with dl-2- (tert-butoxycarbonyl) -2-phenylacetyl chloride as acylating agent. Chromatography of the reaction product gave the titled diester (63.6%) which was crystallized from methylene chloride-hexane to give a white solid (m. P. 184-185 ° C): IR (CHC13) 1801 ^cm-1 (Β-lactam C = O), NMR (CDCl 3) 1.47 (s, 9, t-butyl), 3.27 (s, 3, CO 2 CH 5), 3.40 3.80 (ABq, 2.1) 18.0 Hz, C2-H), 4.45, 4.50 (2-s, 1, sidechain CH), 4.93, 4.96 (2-d, 1, J = 4.0 Hz,

C 6 H, 5.87 (q, 1, J = 5.0 and 9.0 Hz, C 7 -H), 7.10 (s, 1, CH 2), 7.44 (Ar), 7.29 and

8.15 ppm (2-d, 1, J = 9.0 Hz, NH).

Analysis for C35H34N2O8S:

C, 65.41; H, 5.33; N, 4.36. Found C 65.26, H 5.55, N 4.19.

Example 21

7- (2-Phenyl-2-carboxyacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid disodium salt

Benzhydryl-7- (2-phenyl-2-tert-butoxycarbonylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylate (0.257 g, 0.399 mmol) was dissolved in 40 mL of 9.7-109% formic acid and allowed to react one hour at room temperature. The reaction mixture was evaporated to dryness to give a residue which was dissolved in ethyl acetate and extracted with sodium bicarbonate solution. The aqueous extracts were combined, overlaid with cold ethyl acetate and acidified with 1N acid. chtorovoíffieoviMi. The organic phase was separated, washed with brine and dried over anhydrous sodium sulfate. Evaporation to dryness yielded 0.209 g of acid which was dissolved in 40 ml of ethanol and treated with 2 equivalents of sodium 2-ethylhexanoate. The solution was evaporated in vacuo until a precipitate formed. The solution was placed in a refrigerator overnight and then 27 mg of the disodium salt was filtered. NMR (acetone-del 3.75 (m, 5, 10aCH 3, G 2 -H)), 4.85 (2s, 1, sidechain CH), 5.17, 5.28 (2-d, 1, s); 5.0 Hz, GB-1H), 5.92 ppim (q, 1H) = 5.0 Hz and 9.0 Hz, G7-H). . :

Note: doubling of some strips is due to the 'dl-mixture'.

Example 2 2

Benzyryl-7-D - ((2-phenyl-2-t-butoxycarbonylamino) acetamido] -3-methoxycarbonyl-3-cephem-4-carboxylate

To a cooled (2020 ° C), stirred solution of Nt-butoxycarbonylphenylglycine (0.222 g, 0.882 mmol) in 30 mL of tetrahydrofuran was added N-methylhynophenol (0.089 g, 0.882 mmol) under argon, followed by methyl chloroformate (0.097 g, 1.03 mmol). The mixture was allowed to react for 10 minutes at -20 ° C, then the mixture was cooled to -30 ° and a solution of benzhydryl-7-amino-3-methoxycarbonyl-3-cephem-4-carboxylate (0.312 g, 0.735 mmol) was added dropwise. in 8 mL of tetrahydrofuran. The mixture was allowed to react at -25 ° C to -5 ° C for 30 minutes. Cold ethyl acetate was then added, the mixture was washed with cold 1N hydrochloric acid solution, cold sodium bicarbonate solution, brine and dried over anhydrous sodium sulfate. Evaporation and chromatography on silica gel using a gradient of toluene and ethyl acetate afforded 0.305 g (63%) of the title compound: IR (CHCl) 1800 ^cm-1 (1 / J-lactam C = 0) NMR fGDClsj 1.42 (s, 9, tert-butyl], 3.25 (s, 3, CO 2 CH 3), 3.26, 3.75 (ABq, 2, J = 18.0 Hz, C 2 H), 4.85 (d, 1, J = 5.0 Hz), 5.32 (d, 1, J = 6.0 Hz, side chain CH and Cs-H), and 5.82 ppm (m, 2; Ct-H and NH).

1 & W8?

When the aminoester is reacted with N-tert-butoxycarbonylglycine in the presence of a condensing agent (EEDQ), the above product is isolated in 42% yield. Example 23

7-D- (2-Phenyl-2-aminoacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid

Benzhydryl-7-D - [(2-phenyl-2-tert-butoxycarbonylamino) acetamido] -3-methoxycarbonyl-3-cephem-4-carboxylate (0.168 g, 0.256 mmol) was dissolved in 95 mL of 97-100% formic acid and the mixture was stirred at room temperature for 30 minutes. Evaporation in vacuo to dryness gave a product which is insoluble in CDCl 3 and acetone-de. Dissolve the substance in 3 mL cold trifluoroacetic acid. The mixture was stirred in an ice bath for 10 minutes, diluted with n-heptane and evaporated to dryness. The product was dissolved in acetonitrile (9 mL), 1 mL of water was added and the pH was adjusted to 4.5 with dilute sodium hydroxide solution. No precipitate is ruled out. Freeze-drying yields a crystalline product which crystallizes after addition of two drops of water, 5 drops of acetonitrile, 4 drops of water and 4 ml of acetonitrile.

Example 1 a d 2 4

Benzhydryl-7-amino-3- (4-nitrobenzyloxycarbanyl) -3-cephem-4-carboxylate

Using the corresponding 7-thienylacetamido compound as starting material, the amide side chain was cleaved as described in Example 13. The amine ester obtained as product was isolated in 89% yield: IC (CHCl 3) 1799 cm -1 (β-lactam C = O), NMR ( CDCI3)

2.80 (broad singlet, NH 2), 3.74 (center ABq for C 2 -CH) and 4.85 ppm (m, 4, C 6 -H, G 7 -H and CH 2 ester).

Example 25.

!

Benzhydryl-8- (D- (2-phenyl-2-formyloxyacetamido)) - 3- (4-nitrobenzyloxycarbonyl) -3-cephem-4-carboxylate

To a cooled (5 ° C), stirred solution of benzhydryl-7-amino-3- (4-nitrobenzyloxycarbonyl) -3-cephem-4-carboxylate (0.305 g, 0.559 mmol) in 25 mL of tetrahydrofuran was added sodium bicarbonate (0.052). G,

0.514 mmol) was added followed by D-2-phenyl-2-formyloxyacetyl chloride (0.122 g, 0.615 mmol). The mixture was allowed to react with cooling for 30 minutes. Ethyl acetate was added, the mixture was washed with cold water, and with sodium chloride solution, and dried over anhydrous sodium sulfate. Evaporation to dryness and chromatography of the product on silica gel with toluene-ethyl acetate gave 0.183 g (46.4%) of the acylated benzhydryl: IR (CHCl3) 1802 cm @ ^-1 (.beta.-lactam C = Ό) NMR (CDCl3)

3.37, 3.88 (ABq, 2, J = 18.0 Hz, C2-H), 4.67,

4.85 (ABq, 2, J = 14.0 Hz, p-nitrobenzyl CH2), 4.88 (d, 1, J = 5.0 Hz, C5-H), 5.75 (q, 1, J = 5.0 and 9.0 Hz, C7-H), 6.18 (s, 1, side CH), and 8.05 ppm (s, 1, CHO).

Example 1 a d 2 6

7- [D- (2-phenyl-2-formyloxyacetamido)] -3- (4-nitrobanzyloxycarbonyl) -3-cephem-4-carboxylic acid

Following the procedure described in Example 4 wherein the benzhydryl group was removed by treatment with trifluoroacetic acid, the corresponding acid was obtained. The product crystallized from acetone / hexane to give 0.081 g (65%) of tan crystals (m.p. 154-155 ° C).

Analysis for C24H19N5O10S:

calculated C 53.24, H 3.54, N 7.76;

Found C 53.28, H 3.68, N 7.47.

Example 27

Benzhydryl-7 [D- (2-phenyl-2-formyloxyacetamido)] -3-carboxy-3-cephem-4-carboxylate

Benzhydryl-7- [D- (2-phenyl-2-formyl-oxyacetamido)] -3- (4-nitro-benzyloxycarbonyl) -3-methyl-4-carboxylate (0.224 g, 0.3 mmol) was dissolved in 5 ml of methylene chloride and 40 ml of methanol. The re-reduced 5% Pd / C catalyst (0.224 g) was added and hydrogenation was carried out at room temperature (50 psi hydrogen pressure) for two hours. Filtration of the reaction mixture and evaporation of the filtrate to dryness afforded the title compound (190 mg). The product shows antimicrobial activity on gram-positive and gram-negative microorganisms,

SUBJECT

Claims (4)

A process for the preparation of cephalosporin compounds of general formula I, wherein Z is a group of the formula wherein X is a group of formula OR 3 wherein R 3 is alkyl of 1 to 6 carbon atoms or diphenylmethyl, or χ is a group of formula -SR 4 where R 4 is phenyl or benzyl, or X is an azidocupl or a group of formula VYNALEZU H AND R '‘- ΟΙ E kde R "‘ is R "as defined above 2-thienyl or 3-thienyl, E is hydroxyl or hydroxyl protected by alkanoyl having from 1 to 4 carbon atoms, a carboxyl group or a carboxyl group protected with an alkyl of 1 to 6 carbon atoms, amino group or protected alkoxycarbonyl ^one having 1 to 6 carbon atoms in the alkoxy, or R 'denotes heteroarylmethyl of the formula R "" CH2here R '' is 2-thienyl, 3-thienyl, 2-furyl or 3-furyl, and when R is a hydrogen atom, a pharmaceutically acceptable non-toxic acid salt, characterized in that the compound of formula (II) wherein: R 5 is hydrogen or alkyl of 1 to 6 carbon atoms and Q is hydrogen or alkyl of 1 to 6 carbon atoms, wherein: R is hydrogen, benzhydryl or alkyl of 1 to 6 carbon atoms and wherein R 1 is hydrogen or acyl of the formula O II R‘ — C— where R 1 is, arylalkyl of the formula R '~ (Y) m - CH 2 - where R 1 is phenyl or substituted phenyl wherein the substituents are 1 to 3 halogen atoms, hydroxyl, nitro or C 1 -C 4 alkyl, Y is O or S and m is 0 or 1, or R 1 represents a substituted arylalkyl of the formula wherein R 10 is R 1 as defined above, R 1 is a carboxyl protecting group a Z * is a group of the formula wherein A is a bromine, chlorine atom or a mixed anhydride radical of formula O-O-C-O-alkyl wherein alkyl is alkyl of 1 to 6 carbon atoms, is reacted with a compound of formula X — kde where 1952 ⁵⁷ X is as defined for Formula I above and T is hydrogen or an alkali metal, and optionally reacting the corresponding 2-cephem derivatives with a percarboxylic acid to give 3 -cephem derivatives and the resulting sulfoxide is reduced with phosphorus halide and optionally 7-acyl is cleaved by reaction with phosphorus pentachloride and alcohol and optionally re-acylated at the 7-position by reactions With a halide or active ester of an acid R'-CO 2 H, wherein R 'is as defined above, optionally de-esterifying the 3-carboxylic acid ester and / or the 4-carboxylic acid ester in an acidic medium and recovering the product optionally in the form of a 4- carboxylic acid, benzhydryl or C1-C6 alkyl ester of 4-carboxylic acid.