CS195288B2 - Method of preparing cephalosporanic compounds - Google Patents

Description

The invention relates to a process for the preparation of esters of cephalosporin compounds containing a carbonyl group at the 3-position of the dihydrothiazine ring of the cephem cyclic system:

The invention relates to a process for the preparation of compounds of formula I

R is hydrogen or alkyl of 1 to 6 carbon atoms or benzhydryl and R 1 is hydrogen or acyl

O

ti) wherein Z is a group of the formula

wherein R‘ is an arylalkyl of the formula

R "(Y) _{m -} CH _{2 -} wherein R" 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

H í

R '‘- ΟΙ

E wherein R '' is R 'as defined above, 2-thienyl or β-thienyl, E is hydroxyl or hydroxyl protected with 1-4 alkanoyl wherein X is alkyl of 1-6 carbon atoms, methoxybenzyl, nitrobenzyl, benzyl or diphenylmethyl,.

C 1 -C 6 -alkyl, carboxyl or carboxyl-protected, C 1 -C 6 -alkoxy-amino or amino-protected C 1 -C 6 -alkoxy;

R * denotes heteroarylmethyl of the formula R "" CH 2 - wherein 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, the compound of formula (II),

where R 1 is R 1 as defined above,

R 1i is a carboxyl protecting group and

Z ‘is a group of formula ........

COOH is reacted with a compound of formula (I).

X-T wherein X is as defined above for Formula I and T is a diazo group, and optionally reacting the corresponding 2-cephem derivatives with a percarboxylic acid to give 3-cephem derivatives and the resulting sulfoxide is reduced with phosphorous halide and optionally 7-acyl is cleaved with phosphorus pentachloride and an alcohol and optionally re-acylated at the 7-position by reaction with a halide or active ester R'-CO 2 H, where R 'is as defined above, optionally de-esterifying the 3-carboxylic acid ester and / or the 4- and the product is optionally isolated in the form of a salt of a 4-carboxylic acid, a benzhydryl ester or an alkyl ester of 1 to 6 carbon atoms of a 4-carboxylic acid.

A number of cephalosporin series antibiotics are known. All of these antibiotics exhibit the same basic cyclic structure, comprising a four-membered / hlactam ring attached to a six-membered dihydrothiazin ring, and differing in structure from each other with biological effects. Structurally known cephalosporin antibiotics differ from each other in the type of 7-acylamidosubstituent and

also in the substituent type at the 3-position of the dihydrothiazine ring, Desacetoxycephalosporic acid, for example cephalexin, has a 3-methyl substituent. Many known cephalosporins have a substituted methyl group in the 3-position. The desacetylcephalosporins 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 disclosed in U.S. Patent Nos. 3,647,788, 3,668,203 and 3,637,678.

In addition to 3-methyl- or 3-substituted methylephthalosporins, 3-formylisubstituted cephalosporins have also been reported. In U.S. Pat. 1967 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 U.S. Patent No. 3,674,784.

Another group of cephalosporins. distinguishing, in the type of substituent at the 3-position, is disclosed in Dutch Published Application No. 72 06,931, which discloses certain 3-unsubstituted cephalosporins, as well as a process for their preparation by decarboxylation of the corresponding

3-formyl compounds.

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

The term "C 1 -C 6 alkyl" includes methyl, ethyl, n-propyl, n-butyl, isobutyl, pentyl, n-hexyl, cyclohexyl and the like aliphatic, hydrocarbon chains.

Included in the definition of Group X are groups such as. methyl, ethyl, n-propyl, isopropyl, n-butyl, p-methoxybenzyl, benzyl, p-nitrobenzyl, diphenylmethyl and the like.

When in the above definition R 1 is substituted phenyl, R 1 may be mono- or disubstituted haloigenphenyl such as 4-chlorophenyl, 2,6-dlchlorophenyl, 2,5-dlchlorophenyl, 3,4-dlchlorophenyl, 3-chlorophenyl, 3-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. a mononitrophenyl group such as 3- or 4-nitrophenyl, a mono- or disubstituted alkyllenyl group such as 4-methylphenyl, 2,4-dimethylphenyl, 2-methylphenyl, 4-isopropylphenyl, 4-ethylphenyl, 3-n-propylphenyl and the like. R 4 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 carboxy groups 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 to readily cleave by hydrolysis or hydrogenolysis to the corresponding carboxylic acid. Examples of carboxylic acid protecting groups are tert-butyl, benzyl, p-methoxybenzyl, C 2 -C 6 alkanoylmethyl, (3-iodoethyl, p-nitrobenzyl, diphenylmethyl (benzhydryl), phenacyl, p-halophenacyl, 2,2,2-trichloroethyl and the like ester-forming moieties The type of these ester-forming groups is not critical as long as the ester formed is stable under the reaction conditions described below The preferred carboxyl protecting group is benzhydryl, p-hitrobenzyl, tert-butyl , .beta.-trichloroethyl, p-methoxybenzyl and .beta.-iodoethyl, most preferred are 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. McOmie, "Protective Groups in Organic Chemistry", Plenum Press, 1973, is also suitable.

/ G

Examples of acyl groups R 1 -G are benzoyl,

4-chlorobenzoyl, 4-methylbenzoyl, 3,4-dichlorobenzoyl, 3-bromobenzoyl, 3-aminobenzoyl, 4-nitrobenzoyl and the like.

O.....

- <.? ^? Examples of acyl groups R'-C-, wherein R 'is a group of the formula R' - (Y 1 _m -CH 2 -am is O, are phenylacetyl-1,4-chlorophenylacetyl, 3-hydroxyphenylacetyl, 4-hydroxy-3-methylphen acetyl, 4-bromophenylaethyl, 4-nitrophenylacetyl, and the like, and when m is 1 and Y is oxygen, the acyl groups are phenoxyacetyl, 3-hydroxyphenoxyacetyl, 4-chlorophenoxyacetyl, 3,4-dichlorophenoxyacetyl, 2-chlorophenoxyacetyl; 3,4-dimethylphenoxyacetyl, 4-šopropylfenoxyacetyl and ^l, 3-nitrophenoxyacetyl, and the like substituted phenoxyacetyl, and when m is 1 and Y is S, are examples fenylthioacetylskupin fenylthloacetyl, 2,5-dichlorfenylthioacetyl, 3-chloro-4-fluoro-fehylthioacetyl , 3-bromophenylthioacetyl and the like acyl groups.

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

H

R 1 - C - 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 2-hydroxy-2- (3 _: chloro-4-hy, droxyphenyl) acetyl, 2-formyloxy-2- (4-hydroxyphenyl) acetyl, 2-hydroxy -2- (3-bromophenyl) acetyl, 2-formyl-oxy-2- (3,5-dichloro-4-hydroxyphenyl) acetyl, 2-hydroxy-2- (3-chlorophenyl) acetyl and the like: Examples of acyl groups wherein R 'is carboxyl or alkoxycarbonyl substituted arylalkyl, is 2-phenyl-2-carboxyacetyl, 2-phenyl-2-tert-butoxycarbonylacetyl, 2- (4-elilorphenyl) -2.-benzyloxycarbonylacetyl, 2- (3-nitrophenyl) -2-carboxyacetyl and the like.

When R 'is an amino-substituted arylalkyl group or a derivative thereof, the acyl groups are 2-amino-2-phenylacetyl, 2-phenyl-2-tert-butoxycarbonylamino) aceityl, 2- (4-hydroxyphenyl) -2-aminoacetyl and the like acyl groups. ...

O ···; '. ·

Examples of acyl groups R‘-C-, where R‘ is a heteroarylmethyl group of the formula R‘‘-CH 2 -, are 2-thienylacetyl, 3-thienylacetyl, 2-furylacetyl.

Examples of diesters of 7-acylaminocephem-3 ', 4-dicarboxylic acid which are available according to the invention are:.

benzhydryl-7- (2-thiazolylacetamido) -3- (2-bromoethoxycarbonyl) -2-cephem-4-carboxylate,

4‘-nitrobenzyl-7-phenoxyacetamido-3-methoxycarbonyl-3-cephem-4-carboxylate-1-oxide,; tert-butyl 7-acetamido-3-ethoxycarbonyl-3-cephem-4-carboxylate, benzhydryl 1-7-phenylacetamido-3- (2-bromo: propoxycarbonyl) -3-cephem-4-carboxylate,

4‘-Nitrobenzyl-7- (3-thienylacetamido) -3 8

8 7

- (3-bromopropoxycarbonyl) -2-cephem-4-carboxylate,

4'-Methoxybenzyl-7- (2-thienylacetamido) -3- (2-bromoethoxycarbonyl) -3-cephem-4-carboxylate, (3 ', 3', 3'-trichloroethyl 7- (4-chlorophenoxyacetamido) -3- (3-bromo-2-butoxycarbonyl) -2-cephem-4-carboxylate,

2'-iodoethyl-7- (3-furylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylate and tert-butyl 7- (2-thienylacetamido) -3- (2-bromoethoxycarbonyl) -2-cepein -4-carboxylate.

Compounds of formula I may be prepared by esterification of the corresponding 3-carboxylic acids with diazo compounds such as diphenyldiazomeithane or p-nitrophenyldiazomethane. For example, cephem derivatives having a 3-methoxycarbonyl group are prepared by esterifying the 7-acylamino-3-carboxy-3- (or

2-) cephem-4-carboxylic acids, for example diazomethane. Other C-3 carboxylic acid esters, for example trichloroethyl ester, p-methoxybenzyl ester or benzyl ester, can be prepared by conventional esterification techniques and procedures known in the art of cephalosporins, as well as in the fields of general organic chemistry.

7-Methoxy compounds are prepared from 7-methoxy starting materials, for example, from the corresponding 7-methoxy-3-carboxy-3-cephero-4-carboxylic acid esters, '

Removal of the ester protecting group yields novel antibiotic compounds which can be used against infections caused by gram-positive and gram-negative microorganisms.

The cephem-3,4-dicarboxylic acid diester can be converted directly to the biologically active compound by: a) conversion to the 3-cephem derivative (if the diester is originally 2-cephem; bj by cleavage of the C-4-carboxylic acid protecting group ester group).

Rearrangement of the 2-cefemderivatives to the corresponding

The 3-cephem compound is carried out by an oxidoreduction method well known in the chemistry of cephalosporin. Generally, this process is carried out by first oxidizing the 2-cetem compound, for example m-chloroperbenzoic acid, to give the corresponding 3-cefem-1-oxide derivative, which is then reduced with a phosphorus trivalent compound such as phosphorus tribromide or phosphorus trichloride, preferably using dimethylformamide. as 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 or amino group. So when applied to the preparation

8 particularly active compounds are conveniently converted for 2-cephem intermediates with an acylamino group f with each in the acyl, optionally present carboxy or amino protected group] at the C-7 position with the carboxy-protected ester at the 4-position and the ester at the C-3.1 position. in the reaction sequences is not the same, then, if conditions allow, it is experimentally most advantageous to carry out the rearrangement until the C-3 substituent corresponds to the final product.

cleavage of the ester moiety of the molecule at C-4 to a 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, usually in the presence of a cationic ion stabilizer such as anisole. Deesterification of the N, N-trichloroethyl and 2-iodoethyl esters is carried out by reaction with zinc and an acid such as formic acid, acetic acid or hydrochloric acid. The cleavage of the .beta.-nitrobenzyl ester protecting group is usually carried out by hydrogenating the ester in the presence of palladium, rhodium and the like, in suspension or on a carrier such as barium sulfate, coal, alumina, and the like. groups that may be present at any point in the cephem compound.

Cleavage of the ester protecting group carboxy. The 4-position of the 3-cephemediester as defined above leads to the formation of 7-acylamino-3-alkoxycarbonyl (or haloalkoxycarbonyl) -3-cep-4-carboxylic acids, which are preferred antibacterial compounds of the invention.

Examples of such compounds are:

7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephen-4-carboxylic acid,

7-Phenylacetamido-3- {2-bromoethoxycarbonyl] -3-cephem-4-carboxylic acid,

7-phenoxyacetamido-3- (2-iodoethoxycarbonyl) -3-cephem-4-carboxylic acid,

7- (2,5-dichlorophenylthioacetamido-3- (2-bromopropoxycarbonyl) -3-cephem-4-carboxylic acid),

7-chloroacetamido-3-ethoxycarbonyl-3-eephem-4-carboxylic acid,

7-benzamido-3-bromoethoxycarbonyl-3-cephem-4-carboxylic acid,

7- (3-thienylacetamido) -3- (2-iodopropoxycarbonyl) -3-cephem-4-carboxylic acid and (2-thiazolylacetamido) -3-ethoxycarbonyl-3-cephem-4-carboxylic acid.

⁹ . The preparation of the starting materials, 7-acylamino-3-carboxy-3-stero-4-carboxylic acid ester, can be summarized as follows:

1. Preparation of a 7-acylamino-3-formylphenyl-4-carboxylic acid ester;

2. preparation of the acetal derivative (cyclic or acyclic) of the 3-formyl functional group,

3. acetal conversion to the corresponding cephem-3,43-dicarboxylic acid diester by reaction of acetal with N-bromosuccinimide in the presence of a free radical initiator in an inert organic solvent, optionally from 20 to 100 ° C, and

4. deesterification of the ester function

3-carboxylic acid to give the corresponding cephem-3-carboxy-4-carboxylic acid ester.

The process for the preparation of 3-carboxycephemic starting materials can be carried out using materials having a side chain preferred for the preparative process (due to availability or stability to the reaction conditions) and then this side chain can be replaced with other 7-acylamine side chains, preferred for maximum biological activity, using conventional cleavage and reacylation procedures. The intermediates in these transacylations are the corresponding 7-aminocephem derivatives.

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. A preferred base for forming the potassium salt is potassium 2-ethylhexanoate. Salts can be converted ^on free acid 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. The 3 '- (substituted) carbonyl-3-cephem compounds can be formulated into 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 the patient's body weight, disease and other factors related to the patient. Infection can 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 the free acids or in the form of non-toxic salts, such as the sodium or potassium salt.

A very preferred class of active compounds are those of the formula

COOH

O

Wherein R 1 is an acyl group of formula R 1 -C 1 and R 1 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 • Ί

Rio — C'—

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

7- (D-mandelaimido) -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-cyclohexyloxycarbomyl-3-cepine-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.

A n i i n i o n t i o t tests'

The compounds of formula I were tested for activity against some microorganisms to determine the antimicrohial effect. Tests against the organism identified as X-12, a Gram-positive organism of Bacillus subtilis and Serratia, were used as tests.

193288 marcescens, a strain called X-99, which is a gram-negative organism.

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.

Test results for typical compounds of Formula I are as follows:

X-99 X-99

7- (2-thienyl lacetamido) -3-cephem-3,4-di- carboxylic acid 34 mm 7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid 41 mm 7- (2-Thienylacetamido) -3-carboxy-3-cephem-4H-carboxylic acid methyl ester 21 at 10 mg / ml 7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephem-4-fearboxylic acid sodium salt > 200 µg / ml 7- (2-phenyl-2-formyl-oxyacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid, sodium salt 16.5 7- (2-phenyl-2-carboxyacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid disodium salt 33 mm 1 7- [D- (2-phenyl-2-formyloxyacetamido)} -3-methoxycarbonyl-3'-cephem-4-carbaxylic acid 40 7- (2-Phenyl-2-phenyloxy-acetamido) -3- (4-nitro-enoxycarbonyl) -3-cephem-4-carboxylic acid 42 7 7- (2-Phenyl-2-phenyloxyacetamido) -3-cephem-3,4-dicarboxylic acid 130 7- (2,5-dichlorophenylthioacetamiido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid 41 22nd 7- (D- (2-Phenyl-2-formyloxyacetamido)) -3-carboxy-3-cephem-4-carboxylic acid benzhydryl ester 21 at 5 mg / ml 7- (2-phenyl-2-carboxyacetamido) -3-cephem-3,4-dicarboxylic acid 20 May 7- (2-thietylacetamido) -3- (2-bromoethoxycarbonyl) -3-cephem-4-carboxylic acid 31 7- (D- (2-phenyl-2-formyloxyacetamido)] -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- nitrobenzyloxycarbanyl) -3. -cephem-4-carboxylic acid

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

The invention is illustrated in more detail in the following examples.

Preparation!

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

To a solution 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 mL). mol) in 50 mL of ethyl acetate. The reaction mixture was heated to reflux for 15 minutes, cooled to room temperature and evaporated to dryness in vacuo. The residue was washed with 1 L of a 1: 1 mixture of ethylene ether and petroleum ether to give a pink solid: benzhydryl 1-7- (2-thienylacetamide) -3-hydroxymethyl-2-oephem-4-carboxylate (33 g, yield 94.2) %).

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. Isopropyl alcohol (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 400 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 to give 31.3 g (95.4%) of crude benzhydryl 7- (2-thienylacetamido) -3-ortho 1-2-cephem-4-carboxylate which was purified either by crystallization from toluene ¹ (43% yield), 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): IC (CHCl 3) 1785 (/ Malktam C = 0), 1680 (amide C = 0) and 2830 cm ^-1 (forms) 1 C = 10), 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 (8, 1, 4 4 -H) and 9.20 ppm (s, 1, CHO).

Analysis for C27H2ZN2O5S2:

calculated C 62.53, 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-cephem-4-carboxylate

Benzhydryl-7- (2-thienyl lacetamido-3-formyl-2-cephem-4-carboxylate (21.5 g, 41.5 mmol) was treated with 11.6 mL ethylene glycol (0.2 mol) and monohydrate p. toluene-sulfonic acid (0.197 g, 1.04 mmol) in 500 mL of 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 mixture was extracted with ethyl acetate and washed successively with sodium bicarbonate solution (2X), water (2X) and brine, and then dried over sodium sulfate, and evaporated in vacuo to dryness to give a product which was chromatographed on 40 g of silica gel using a benzene / ethyl acetate gradient. Crystallization of the purified product from methylene chloride and hexane gave benzhydryl-7- (2-thienylacetamido) -3- (1,3-dioxolan-2-yl) -2-cephem-4-carboxylate as colorless needles (15.07 g). Mp 142-143 ° C, IR (CHCl 3) 1780 cm -1 (O-lactam C = O), NMR (CDCl 3) 3.3-3.9 (m, 4, CH 2) —CH2—), 3, 83 (s, 2, CH 2 sidechain), 5.10 (d, 1, J = 4.0 Hz, C 6 -H), 5.17 s, 1, acetal CH), 5.21 (s, 1 , C1-H) and 5.45 ppm (q, 1, J = 4.0 and 8.0 Hz, C7-H). Analysis for C29H26O6S2:

calculated C 61.69, H 4.66, N 4.98;

found, C 61.69, H 4.43, N 5.10.

Preparation 3

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

Benzhydroxy-7- (2-thienylacetamido) -3- (1,3-dioxolan-2-yl) -2-cephem-4-carboxylate (15.07 g, 25.8 mmol) was mixed with N-thrusuccinimide (5.25 g, 29.5 µmol) and azobisisobutyronitrile (36.5 mg, 0.25 mmol, 0.01 eq) in 1200 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 toluene / ethyl acetate gradient gave 7.61 g (44.4%) of benzhydryl-7- (2139288).

thienylacetamido) -3- (2-bromoethoxycarbonyl) -2-cephem-4-carboxylate, mp 129-130 ° C, IR (CHCl 3) 1785 cm -1; ([Beta] -lactam C = O), NMR (CDCl3) 3.25 (t, 2, J = 6.0 Hz, CH2 Br), 3.83 (s, 2, CH2 side chain),

4.30 (t, 2, J = 6.0 Hz, O-CH 2 -), 4.95 (d, 1, J = 4.0 Hz, C 6 -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 C29H25BrN2OeS2:

calculated: C 54.29, H 3.93, N ¹ 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-bromomethoxycarbonyl) -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 to yield 7.78 g (95.5%) of benzhydryl 7- (2-thienylacetamido) -3- (2-iodoethoxy ^l carbonyl) -2-cephem-4-carboxylate: IR (CHCl) 1785 cm -1 ^-1 (N-lactam C = O), NMR (CDCl 3) 2.96 (t, 2, J = 7.0 Hz, CH 2), 3.80 (s, 2, CH 2 sidechain), 4 24 (t, 2, J = 7.0 Hz, -OCH 3 -), 4.95 (d, 1, J = 4.0 Hz, Ca-H), 5.24 (q, 1, J = 4.0 Hz, C7-H, remainder of the signal overlaps with Ca-H), 5.50 (s, 1, Cl), and

7.80 ppm (s, 1, Ca-H).

5

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

Benzhydryl-7- (2-thienylacetamido) -3- (2-iodoethoxycarbonyl) -2-cephem-4-carboxylate (2.79 g, 4.05 mmol) was dissolved in a mixture of 8 mL of glacial acetic acid and 48 mL of dimethylformamide at 0 ° C and the lever is 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 afforded 1.92 (89%) of benzyryl 7- (2-thienylacetamido) -3-carboxy-2-cephem-4-carboxylate NMR (CDCl 3) 3.84 (s, CH 2 sidechain),

4.99 (d, 1, J = 4.0 Hz, C 6 -H), 5.45 (m, C 4 -H and C 7 -H) and 7.80 ppm (s, C 2 -H).

IB

He did

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

A solution of diazomethane (0.69 g, 16.2 mmol) in ethyl acetate was added dropwise to a stirred cooled (about 5 ° C) solution of benzhydryl-7- (2-thienylacetamido) -3-carboxy-2-cephem-4-carboxylate (5). (80 g, 10.8 mmol) in 250 mL of ethyl acetate. After 30 minutes the excess diazomethane was quenched with acetic acid. The reaction mixture was washed with sodium bicarbonate solution, water and brine, then dried over anhydrous Na ² SO 4. Evaporation of the ethyl acetate solution in vacuo gave 5.72 g (96.3%) of benzhydryl-7- (2-thienylacetamido) -3-methoxycarbonyl-2-cephem-4-carboxylate (TLC): NMR (CDCl3 and 3.67 (s, CO2CH3), 3.84 (s, CH2 side chain), 5.00 (d, J = 4.0 Hz, Ce-H), 5.00 (m, J = 4.0 and 8.0 Hz, C7-H) and, 7.66 ppm (s, C2-H).

Example 1 a d 2 t

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 the cooled (ice bath for 10 minutes), stirred solution of benzhydryl-7- (2-thienylacetamido) -3-methoxycarbonyl- Of 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-cephemesulfoxide (thin layer chromatography - one spot). The product was dissolved in 100 ml diraethylformamide (cooled in an ice bath) and then 1.37 ml phosphorous trichloride (15.7 mmol, 1.5 eq) was added. 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 sodium 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 proc.) Of benzhydryl-7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylate as a yellow solid, mp 201 ° C, rodei, (methyl chloride-hexane) ), UV max (EtOH) 283 nm (ε 13.6 X 10 ³ ), inflex 243 nm, IR (CHCl 3) 1798 cm -1 [η-lactam C = O], NMR (DMSO-d 6) 3.30 (s, 3, CO2CH3), 3.74, 3.80 (m, 4, CH2 side chain, C2-H), 5.25 (d, 1, J = 5.0 Hz, C5-H), 5 90 (q, 1, J = 4.0 and 8.0 Hz, C2-H) and 7.0, 7.4 ppm (im, 13, benzhydryl ester, thlenyl).

Example 1 a d 3

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

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

To the cooled (5 ° C) suspension. benzhydryl-7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylate (0.316 g, 0.579 mmol) in 5 mL of anlsole was added 5.0 mL of cold trifluoroacetic acid. The colorless solution was stirred for 10 minutes with cooling, then 50 ml of n-heptane was added and the resulting solution was concentrated to a small volume in vacuo to precipitate a white solid, which was filtered off and then dissolved in acetone. 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. 0.214 g (97%) of 7- (2-thienylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid are obtained in vacuo to dryness.

Example 1 a d 4

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

To a stirred solution of benzhydryl-7- (2-thienylacetamido) -3-carboxy-2-cephem-4-carboxylate (1.604 g, 3 mmol) in 175 mL of ethyl acetate was added dropwise 65 mL of a solution of 4-nitrophenyldiazomethane (0.013 g / mL, 5.18 mmol). The reaction mixture was stirred at room temperature for 48 hours and then evaporated to dryness in vacuo. The product is chromatographed on silica gel using a gradient of toluene and ethyl acetate. 1.66 g (83%) of behzhydryl-7- (2-thienylacetamido) -3- (4-nitro-benzyloxycarbonyl) -2-cephem-4-carboxylate are obtained in the form of a yellow foam, IC (CHCl3) 1788 cm ^-1. (R-lactam C = O), NMR (CDCl 3) 3.82 (s, 2, side chain CH 2), 4.95 (d, 1, J = 4.0 Hz, C 6 -H), 5.12 (S, 2, nitrobenzyl CH2), 5.45 (q, 1, J = 4.0 and 8.0 Hz, C7-H), 5.50 (s, 1, Ca-H),

7.80 (s, 1, C2-H), and 6.9, 7.3 ppm (ArH). Example 5

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

Using the procedure described in Example 2, except that benzhydryl-7- (2-thienylacetamido) -3- (4-nitrobenzyloxycarbonyl) -2-cephem-4-carboxylate was used in place of the corresponding methoxycarbonyl compound. cephem compound was isolated in 48% yield and crystallisation from methylene chloride, acetone and hexane gave pale yellow needles (mp 165 DEG 1JB6 ° C): IR (CHC) 1801 cm @ ^-1 (.beta.-lactam C = 0] NMR (CDCl3 3.42, 3.85 (ABq, 2, J = 18.0 Hz, C2-H), 3.80 (s, 2, CH2 side chain), 4.54, 4.94 (ABq, 2, J = 12.0 Hz, nitrobenzyl CH2), 4.95 (d, 1, J = 5.0 Hz, Ce-H), 5.85 (q, 1, J = 5.0 and 9.0 Hz, C7-H) and 7.0, 7.2 ppm (ArH).

Analysis for C34H27N3O8S2:

H, 4.06; N, 6.27.

Found: C 61.20, H 4.24, N 6.33.

Example

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

Zinc dust (1.50 g) was added to a cooled, stirred solution of benzhydryl-7- (2-thienylacetamido) -3- (2-iodoethoxycarbonyl) -2-cephem-4-carboxylate (1.60 g, 2.33 mmol) 5 ml of glacial acetic acid and 30 ml of dimethylformamide. The mixture was allowed to cool with stirring for 1.5 hours, then the reaction mixture was diluted with ethyl acetate and filtered through celite. The filtrate was then washed successively with sodium bicarbonate solution (3X), 1N hydrochloric acid, and brine, then dried over anhydrous sodium sulfate to give the corresponding 3-carboxy starting material. Excess diphenyldiazomethane was added and the mixture was stirred gently for 10 minutes. Evaporation to dryness gave a residue which was chromatographed on a silica gel column using a gradient of benzene and ethyl acetate. 1.289 g (79%) of benzhydryl-7- (2-thieneacetamido) -3-behzhy dry-alkoxycarbonyl-2-cephem-4-carboxylate are obtained: IR (CHCl3) 1785 cm @ ^-1 (R) = C = O) NMR (CDCl 3, 3.90 (s, 2, CH 2 sidechain), 4.94 (d, J = 4.0 Hz,

1, Cs-H), 5.41 (q, 1, J = 4.0 and 8.0 Hz), 5.59 (s, 1, Ca-H) and 7.80 ppm (s, 1, C2) -H). Example 7

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

Carry out the same oxidation-reduction procedure described in Example 2. Chromatography of the crude product on silica gel with benzene-ethyl acetate gave 15.2% of the starting 2-cephem and 3-61.6% cefemdibenzhydrylesteru IR (CHCl3) 1799 CMI ^(: /? - lactam C = O), NMR (CDCl 3) 3.33, 3.84 (ABq,

2, J = 18.0 Hz, C 2 -H), 3.78, (s, 2, lateral

198288 chain CH 2), 4.90 (, d, 1,) === 4.0 Hz, C 6 -H) and 5.85 ppm (q, 1, J = 4.0 and 9.0 Hz, C 7 - H). Example 8

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

Zinc dust (1.573 g, 24 mmol) was added to a cooled (about 5 ° C), stirred solution of methyl 7- (2-thlenylacetamido) -3- (2-iodoethoxycarbonyl) -2-cephem-4-carboxylate (1.573 g) (2.93 mmol) in a mixture of 5 ml of glacial acetic acid and 30 ml of dimethylformamide. The reaction mixture was stirred with cooling

Ethyl acetate was added for 1.5 hours. The resulting mixture was filtered through celite. The filtrate was washed successively with sodium bicarbonate solution (3X), 1N hydrochloric acid, and brine, then dried over sodium sulfate. Evaporation of the reaction mixture to dryness yielded 1.305 g of product which was dissolved in ethyl acetate and treated with excess diphenyldlazomethane. The mixture was heated to reflux for 10 minutes and then cooled to room temperature. Evaporation of the reaction mixture to dryness yielded a product which was chromatographed on silica gel using a gradient of benzene and ethyl acetate. 0.842 g (52.5%) of methyl 7- (2-thienylacetamido) -3-benzhydryloxycarbonyl-2-cephem-4-carboxylate was obtained, NMR 3.58 (s, 3, CO2CH3), 3.78 (s, 2). , side chain CH 2), 5.08 (d, 1, J = 4.0 Hz, C 6 -H), 5.42 (s, 1, C 4 -H), 5.57 (q, 1, J = 4.0 and 8.0 Hz, C7-H) and 7.75 ppm (s, 1, C2-H).

Example 9

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

The same oxidation reduction procedure as described in Example 2 is carried out. The corresponding 2-cep is used as starting material. (62 ° / o), crystallized from methylene chloride-hexane to give white needles (ht 163-164 DEG C IR (1798 cm CHClsJ ¹ (/? - lactam C = 0) NMR (CDCl3) 3.45 3.87 (ABq, 2, J = 18.0 Hz, C2-H], 3.53 (s, 3, CO 2 CH 3), 3.85 (s, 2, CHa side chain), 5.00 (d 1.1 (4.0 Hz, Ce-H) and 5.85 ppm (q, 1, J = 4.0 and 8.0 Hz, C7-H).

Analysis for C28H24N2O6S2:

H, 4.41; N, 5.11.

Found: C 61.07, H 4.33, N 4.86.

Example 10

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

The same side chain cleavage procedure as described in Example 7 was followed. The corresponding 2-thlenylacetamido compound was used as starting material. The product, the cephem compound diester, was isolated in 93% yield (one spot in thin layer chromatography). IR (GHCb) 1795 cm ¹ (, / 3-lacquer == 0 where C) NMR (CDCl3)

1.84 (broad singlet, 2, NH 2, exchangeable with D 2 O), 3.27 (s, 3, CO 2 CH 3), 3.41, 4.90 (ABq,

2, J = 18.0 Hz, C2-HJ, 4.70 (d, 1, J = 5.0 Hz, Ce-H), 4.92 (d, 1, J = 5.0 Hz, C7) -H), 7.11 (s, 1, benzhydryl CH), and 7.4 ppm (s, 10, benzhydryl ArH).

Example 11

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

Following the same acylation procedure as described in Example 8, only benzhydryl-7-amino-3-methoxycarbonyl-3-cephem-4-carboxylate was used as the starting material. Chromatography gave 0.211 g (61.2%) of product which crystallized from a mixture of methylene chloride and hexane as white needles (mp 207-208 ° C): IR (CHCl 3) 1800 cm -1 [β-lactam C = OO, NMR (acetone de)) 3.34 (s,

3, CO 2 CH 3), 3.7 (m, C2-H), 5.15 (d, 1, J = 5.0 Hz, C 6 -H), 5.95 (q, J = 5.0 and 9, 0 Hz, C7-H),

6.30 (s, 1, side chain CH), 7.10 (s, 1, benzhydryl CH), 7.5 (m, 10, benzhydryl ArB) ,. 8.34 (s, 1, CHO), and 8.43 ppm (d, J, J = 9.0 Hz, NH).

Analysis for CstHaSNaOeS:

C, 63.47; H, 4.47; N, 4.78.

Found: C 63.40, H 4.73, N 4.53 '.

Example 12

7- (O- (2-phenyl-2-formoxyacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylic acid) Following the same procedure as in Example

4 wherein the benzhydryl ester is de-esterified by treatment 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 C 16 H 16 N 2 O 6 S:

calculated C 51.43, H 3.84, N 6.66;

Found: C, 51.70; H, 4.06; N, 6.77.

Example 13

Benzhydryl-7 - [(2,5-dichlorophenylthio) acetamide] 3-methoxycarbonyl-3-cephem-4-carboxylate

To a cooled (5 ° C), stirred solution of benzhydryl-7-amino-3-methoxycarbonyl-3-cephem-4-carboxylate (0.268 g, 0.632 mmol) in 30 mL tetrahydrofuran was added 0.239 g (28.4 mmol) sodium bicarbonate. and 0.390 g (1.52 mmol) of 2,5-dichlorophenylthioacetic acid chloride. The mixture was stirred with cooling for one hour. Ethyl acetate was added and the resulting solution was a defect and brine and dried over anhydrous sodium sulfate. Chromatography on silica gel using a toluene / ethyl acetate gradient gave 0.288 g (71%) of the title compound as a white solid, which crystallized from methylene chloride / hexane to give 173 mg colorless crystals (mp 179-180 ° C), IC (CHCl 3). 1805 cm ^-1 , NMR (CDCl 3, acetone d-6) 3.30 (s, 3, CO 2 CH 3), 3.50, 3.85 (ABq, 2, J = 16 Hz, C 2 -H),

O f

with

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

Analysis for C30H24N2O6S2Cl2:

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 was prepared by cleavage of the benzhydryl ester with trifluoroacetic acid and anlsole as described in Example 4. The product showed antimicrobial activity.

Example 14

Benzhydryl-7- (2-phthyl-2-tert-butoxycarbonylacetamido) -3-methoxycarbonyl-3-cephem-4-carboxylate

Following the procedure of Example 8, 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 afforded the title diether (63.6%) which was crystallized from: methylene chloride hexane to give a white solid (mp 184-185 ° C): IC (CHCl 3) 1801 em ¹ (0-) lactam C = O), NMR (CDCl 3) 1.47 (s, 9, t-butyl),

3.27 (s, 3, CO 2 CH 3), 3.40, 3.80 (ABq, 2, J = 18.0 Hz, C2-H), 4.45, 4.50 (2-s, 1 , side chain CH), 4.93, 4.96 (2-d, 1, J = 4.0 Hz, Ce-H), 5.87 (q, 1, J = 5.0 and 9.0 Hz) , C7-H),

7.10 (s, 1.0 Hz), 7.4 (Ar), and 7.29, 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 15

7- (2-Phenyl-2-carboxyacetamido) -3-methoxycarbonyl-3-cephem-4-carboxy-, disodium salt; acids

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 97-100% 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 hydrochloric acid. 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 is. evaporate in vacuo until a precipitate formed. The solution was placed in a refrigerator overnight and then filtered to give 27 mg of the disodium salt; NMR (acetone-de) 3.75 (m, 5, CQ 2 CH 5, C 2 H),

4.85 (2-s, 1, sidechain CH), 5.17, 5.28 (2-d, 1, J = 5.0 Hz, Ce-H) and 5.92 ppm (q, -1) J - 5.0 Hz and 9.0 Hz, C7-H).

Note: doubling of some strips is caused by dl-mixture.

Example 16

Benzhydryl-7-D - [(2-phenyl-2-tert-butoxycarbonylamino) acetamido] -3-methoxycarbonyl-3-cephem-4-carboxylate

To a cooled (-20 ° C), stirred solution of N-tert-butoxycarbonylphenylglycine (0.222 g, 0.882 mmol) in 30 mL tetrahydrofuran was added N-methylmorpholine (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 cooled to -30 ° C and a solution of benzhydryl-7-amino-3-methoxycarbonyl-3-cephem-4-carboxylate (0.312 g, 0.735 mmol) in dropwise was added dropwise. 8 ml of tetrahydrofuran. The mixture was allowed to react for 30 minutes at -25 ° C to -5 ° C. Cold ethyl acetate was then added, the mixture was washed with cold 1N hydrochloric acid, cold sodium bicarbonate, brine and dried over anhydrous sodium sulfate. Evaporation and chromatography on silica gel using a gradient of toluene and ethyl acetate gave 0.305 g (63%) of the title compound: IC (CHCl3) 1800 cm @ ¹ (n-lactam C = O), NMR (CDCl3)

1.42 (s, 9, t-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 C 6 -H), and 5.82 ppm (m, 2, C 7 - HaNH).

If the aminoester is reacted with N-tert-butoxycarbonylphenylglycine in the presence of a condensing agent (EEDQJ), the above product is isolated in 421% yield.

Example 17

7-β- (2-phenyl-2-aminoacetamldo) -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) is dissolved in 95 mL of 97-100% acid form and the mixture was stirred at room temperature for 30 minutes. Evaporation in vacuo to dryness yielded a product which is insoluble in CDGl3 and acethone. 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 hydroxide solution. sodium. No precipitate is formed. Frost sublimation yields crystalline. a product which crystallizes after addition of 2 drops of water, 5 drops of acetonitrile, 4 drops of water and 4 ml of acetonitrile.

Example 18

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

Using the corresponding 7-thienylacetamido compound as starting material, the amidino side chain was cleaved as described in Example 7. The amino ester obtained as product was isolated in 89% yield: IC (CHCl 3) 1799 cm ^-1 (β-lactam CO), NMR (CDCl 3)

2.80 (broad singlet, NH2), 3.74 (ABq center for C2-H), and 4.85 ppm (m, 4, C5-H, C7-H and CH2 ester).

Example 19

Benzhydryl-7- [D- (2-phenyl-2-formyl-oxyacetamido)] -3- (4-nitrobenzyloxycarbonyl) -3-cephem-4-carboxylate

To the cooled (5 ° C), stirred solution of benzhydryl-7-amino-3- (4-nitrobenzyloxycarbonyl) -3-cephem-4-carboxylate (0.305 g,

0.559 mmol): Sodium hydrogen carbonate (0.052 g, 0.514 mmol) was added in 25 mL tetrahydrofuran, followed by D-2-phenyl-2-formyoxyacetyl chloride (0.122 g, 0.615 mmol). The mixture was allowed to react with cooling for 30 minutes. Ethyl acetate was added, then the mixture was washed with cold water and brine, dried over anhydrous sodium sulfate, evaporated to dryness and chromatographed on silica gel with toluene / ethyl acetate gradient to give 0.183 g (46.4%) of the acylated benzhydryl ester: IC (CHCl 3). ) 1802 cm ¹ (β-lactam C = O), NMR (CDCl 3)

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

4.85 (ABq, 2, J = 14.0 Hz, p-nitrobehzyl CH2), 4.88 (d, 1, I = 5.0 Hz, C6-H), 5.75 (q, 1, J = 5.0 and 9.0 Hz, C7-H), 6.18 (s, 1, side chain CH) and 8.05 ppm (s, 1, CHO). Example 20 ’

7- [D- (2-phenyl-2-formyl-oxyacetamido)] -3- (4-nitrobenzyloxycarbonyl) -3-cephem-4-carboxylic acid

The procedure described in Example 4 wherein the benzhydryl group was removed by treatment with trifluoroacetic acid gave the corresponding acid. The product was crystallized from acetone / hexane to give 0.081 g (65%) of yellowish brown crystals (mp 154-155 ^° C). Analysis for C24H19N3O10S:

calculated: C 53.24, H 3.54, N 7.76;

Found: C 53.28, H 3.68, N 7.47.

Example 21

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

Benzhydryl-7- [D- (2-phenyl-2-formyl-oxyacetamido)] -3- (4-nitrobenzyl-oxycarbonyl) -3-cephem-4-carboxylate (0.224 g, 0.3 mmol) was dissolved in 5 ml methylene chloride and 40 ml miethananol. Re-reduced with 5% Pd / C catalyst (0.224 g) was added and hydrogenation was carried out at room temperature (hydrogen pressure 0.35 MPa) for two hours. Filtration of the reaction mixture and evaporation of the filtrate to dryness afforded the title compound (190 mg). The product shows anti-mlrobial activity on gnam-positive and gram-negative microorganisms.

Claims (1)

A process for the preparation of cephalosporin compounds of formula I wherein Z is a group of formula wherein X is alkyl of 1 to 6 carbon atoms, methoxybenzyl, nitrobenzyl, benzyl or diphenylmethyl, wherein R is hydrogen, alkyl of 1 to 6 carbon atoms, or benzhydryl and wherein R 1 is a hydrogen atom or an acyl group of the formula (IIa) or an amino group protected by C 1 -C 6 alkoxycarbonyl, or R 'represents a heteroarylmethyl of the formula R "" CH 2 - wherein R "" is 2-thienyl, 3-thienyl, 2-furyl or 3-furyl, and when R is hydrogen, a pharmaceutically acceptable non-toxic acid salt, characterized in that the compound of formula (II), R&NH / Sk COORn (II) wherein R 1 is R 1 as defined above, R 11 is a carboxyl protecting group α Z * is a group of formula COOH wherein R 'is arylalkyl of the formula R "(Y) _m -CH 2 - wherein R" 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 H E wherein 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 carboxy protected by C 1 -C 6 alkyl, the amino-reaction does not react with a compound of formula Wherein X is as defined above for Formula I and T is a diazo group, and optionally reacting the corresponding 2-cephem derivatives with a percarboxylic acid to give the 3-cephem derivatives and the resulting sulfoxide is reduced with phosphorous halide and optionally 7-acyl is cleaved by reaction with phosphorus pentachloride and an alcohol and optionally re-acylated at the 7-position by reaction with a halide or active ester of R'-CO 2 H, where R 'is as defined above, optionally esterifying the 3-carboxylic acid and / or 4-carboxylic acid ester in acidic The medium and the product are isolated, optionally in the form of a salt of a 4-carboxylic acid, a benzhydryl ester or an alkyl ester of 1 to 6 carbon atoms of a 4-carboxylic acid.