BG60384B2 - Cephalosporin antibiotic compositions - Google Patents

Abstract

Methods and intermediate compounds have been established for the production of cephalosporin compounds with substituent in 7th place instead of hydrogen. The new cephalosphorin compounds are active against different gram-negative and gram-positive organisms.

Description

The invention relates to new antibiotics, new intermediates useful for the preparation of these antibiotics, and to a method for the preparation of these compounds. In particular, it relates to novel derivatives of 7-aminocephalosporanic acid with a substituent at position 7 and to novel intermediates and a method for their production. 1

The discovery of penicillin, which has been found to be an important and effective antibiotic, is of great interest to the field. Subsequently, various other 2 antibiotics, such as streptomycin, tetracycline and the like, were found that greatly increase the ability of physicians to treat infections due to various pathogens. But the use of 2 of these antibiotics gives rise to strains of pathogens resistant to them. Known antibiotics also have the disadvantage that they are effective only against certain types of microorganisms and are not effective against a wide range of pathogens. Therefore, the search for new antibiotics continues.

It is an object of the invention to provide novel cephalosporins with antibiotic activity, to provide a method for the preparation of these new antibiotics, and to provide new intermediates useful for the preparation of these new cephalosporins. Further directions are disclosed by the detailed description of the invention given below.

The new cephalosporins according to the invention are compounds wherein 0 Δ ³ is the cepham ring, namely the dehydrothiazine ring with fused β-lactam contains a substituent in the 7th place. The new cephalosporins can be represented by the following 5 structural formula

in which R 1 represents an acyl group, A represents an organic radical or group and R 1 represents a radical or hydrogen substituent group and derivatives thereof, such as esters, amides and salts, are novel antibiotic substances.

The acyl radical represented by the symbol R 'may be a substituted or unsubstituted aliphatic, aromatic, heterocyclic or araliphatic radical, or a heterocyclylaliphatic carboxylic acid radical, or a carboxylic acid radical, such as acylphenyl cephilic radicals. These acyl radicals can be represented by the general formula - CO / CH / / CH, / R.

| GL I L L in which R ₂ is a radical of the group defined below, the type have a value from 0 to 4 and R ₃ represents R or ZR, which are defined below.

A group of acyl radicals may be represented by the general acyl group formula

O II - C - R in which R represents a substituted and unsubstituted, straight or branched, alkyl, alkenyl or alkynyl group; aryl, aralkyl; cycloalkyl; or a heteroaryl or heteroaralkyl group. These groups may be unsubstituted or substituted by radicals such as alkyl, alkoxy, halo-, cyano.-, carboxy-, sulfoamino-, carbamoyl, sulfonyl, azido-, amino-, substituted amino, haloalkyl, carboxyalkyl, carbamoyl, carbamoyl, carbamoyl substituted carbamoylalkyl, guanidino, N-substituted guanidino, guanidinoalkyl and the like. Examples of such acyl groups which may be mentioned are those in which R is benzyl, p-hydroxybenzyl, 4-amino-4-carboxybutyl, methyl, cyanomethyl, 2-pentenyl, n-amyl, n-heptyl, ethyl 3 - or 4-nitrobenzyl, phenethyl, β, β-diphenylethyl, methyldiphenylmethyl, triphenylmethyl, 2-methoxyphenyl, 2,6-dimethoxyphenyl, 2, 4, 6-trimethoxyphenyl, 3, 5-dimethyl-4-isoxazolyl, 3-butyl-5-methyl 4-isoxazolyl, 5-methyl-3-phenyl-4-isoxazolyl, 3- (2-chlorophenyl) -5-methyl-4-isoxazolyl, 3- (2,6-dichlorophenyl) -5-methyl-4-isoxazolyl, O-4-amino-4-carboxybutyl, D-

4- N -benzoylamino-4-carboxy-n-butyl, p-aminobenzyl, o-aminobenzyl, taminobenzyl, (3-pyridyl) methyl, 2ethoxy-1-naphthyl, 3-carboxy-2-quinoxalinyl, 3- (2) 6-dichlorophenyl / -

5- (2-furyl) -4-isoxazolyl, 3-phenyl-

4-isoxazolyl, 5-methyl-3- (4-guanidinophenyl) -4-isoxazolyl, 4-guanidinomethylphenyl, 4-guanidinomethylbenzyl, 4-guanidinobenzyl, 4-guanidinophenyl, 2,6-dimethoxy-4-guanidinophenyl-4-guanidinophenyl , p-carboxymethylbenzyl, p-carbamoylmethylbenzyl, m-fluorobenzyl, t-bromobenzyl, p-chlorobenzyl, p-methoxybenzyl, 1-naphthylmethyl, 3-isothiazolylmethyl, 4-isothiazolylmethyl, 5-isothiazolylmethyl-5-methylsilylmethyl-5-methylsilylmethyl-5-methylsulfonylmethyl -isoxazolylmethyl,

4-methyl-5-isoxazolylmethyl, 1-imidazolylmethyl, 2-benzofuranylmethyl,

2- and idolyl methyl, 2-phenylvinyl, 2-phenylethynyl, 2- (5-nitrofuranyl) vinyl, phenyl, o-methoxyphenyl, o-chlorophenyl, o-phenylphenyl, p-aminomethylbenzyl, 1- (5-cyanotriazolyl) methyl , difluoromethyl, dichloromethyl, dibromomethyl, 1- (3-methylimidazolyl) methyl, 2- or 3- (5-carboxymethylthienyl) methyl, 2- or 3- (4-carbamoylthienyl) methyl, 2- or 3- (5-methylthienyl) methyl, 2- or 3- (5-methoxythienyl) methyl, 2- or 3- (4-chlorothienyl) methyl, 2- or 3- (5-sulfothienyl) methyl, 2- or 3- (5-carboxythienyl, 3) 1,2,5-thiadiazolyl] -methyl, 3- (4-methoxy-1,2,5,5-thiadiazolyl) methyl, 2-furylmethyl, 2- (5-nitrofuryl) methyl,

3-furylmethyl, 2-thienylmethyl, 3-thienylmethyl and tetrazolylmethyl.

The acyl group may also be a radical of the formula

O II - C / CH, / ZR η

in which η has a value from 0 to 4, ζ is oxygen or sulfur and R is defined as above. As examples of the substituent - / CH, / ZR

Allytomethyl, phenylmethyl, phenylmethylenediamine, polyethylene 2-thiazolylmethyl, p- (sulfo) phenoxymethyl, p- (sulfo) phenylthiomethyl, p- (carboxy) phenoxymethyl, p- (carboxy) phenylthiomethyl, p- (carboxymethyl) phenoxymethyl, p- (carboxymethyl) phenylthiomethylthiothiolthiolthiolthiolthiolthiomethyl, 2- , hair dryer ilthiomethyl, 1- (5, 6, 7, 8-tetrahydronaphthyl) oxomethyl, 6, 8-bis (methylthio) octanoyl.

The acyl group may also be a radical of formula

O II - C - CHR

I

R 'in which R has the meanings indicated and R' denotes the radicals amino, -hydroxy, azido-, carbamoyl, guanidino-, acyloxy, halo-, sulfamino-, tetrazolyl, sulfo-, carboxy-, carbalkoxy and the like. As typical examples of substituents for

-CHR IR may indicate α-aminobenzyl, amino-2-tenyl, α-methylaminobenzyl, α-amino-γ-methylmercaptopropyl, amino-3- or 4-chlorobenzyl, α-amino-3- or 4-hydroxybenzyl, and -amino2,4-dichlorobenzyl, α-amino-3, 4-dichlorobenzyl, D (-) -α-hydroxybenzyl,? -carboxybenzyl,? -amino-3-tenyl, amino-2-tenyl, O (-) -? -amino-3-chloro-4-hydroxybenzyl, D (-) -a-amino-3-phenyl, 1-aminocyclohexyl, α- (5-tetrazolyl) -benzyl, α-sulfaminobenzyl, α-sulfamino-3-tenyl, α- ίΝ-methylsulfamino) -benzyl, D (-) -α-guanidino-2-tenyl, D (-) -α-guanidinobenzyl, α-guanylureidobenzyl, α-hydroxybenzyl, α-azidobenzene yl, α-fluorobenzyl, 4- (5-methoxy-1,3-oxydiazole) aminomethyl, 4- (5-methoxy-1,3-oxidiazole) -hydroxymethyl, 4- (5-methoxy-

1,3-Oxidiazole-carboxymethyl, 4- (5-methoxy-1,3-sulfadiazole) -aminomethyl, 4- (5-methoxy-1,3-sulfadiazole) hydroxy-methyl, 4- (5-methoxy-1,3 -sulfadiazole) -carboxymethyl, 2- (5-chlorothienyl) -aminomethyl, 2- (5-chlorothienyl) -hydroxymethyl, 2- (5-chlorothienyl) -carboxymethyl, 3- (1,2-thiazole) aminomethyl, 3- ( 1,2-thiazole) -hydroxymethyl, 3- (1,2-thiazole) -carboxymethyl,

2- (1,4-thiazolyl) -aminomethyl, 2- (1,4-thiazolyl) -hydroxymethyl, 2- (1,4-thiazolyl) -carboxymethyl, 2-benzothienylaminomethyl, 2-benzothienylhydroxymethyl, 2-benzothienylcarboxymethyl,

2-azidoooctyl-3-phenyl-3-azidomethyl, α-sulfobenzyl and α-phosphonobenzyl.

N.

I

The group R '- N - may be sulfonamide, such as phenylsulfonamido, -ethylsulfonamido, benzylsulfonamido, 2, 5-dimethylsulfonamido, 4-chlorosulfonamido,

4-chloro-phenylsulfonamido, 4-methoxysulfonamido and the like.

Acyl substituents of the general formula

R10 is CHCO in which R.0 and R10, as defined below, represent a preferred group of substituents because of their generally useful antibiotic activity. _{R] 0} represents hydrogen, halo, amino, guanidino, phospho, hydroxy, tetrazolyl, carboxy, sulfo or sulfaminogrupa. R _H represents phenyl, substituted phenyl, monocyclic heterocyclic 5- or 6-membered ring containing one or more oxygen, sulfur or nitrogen atoms in the ring, substituted heterocycles, phenylthio-, heterocyclic or substituted heterocycles, phenylthiocylene, heterocyclic, heterocyclic or heterocyclic heterocyclic, thio groups or cyano groups. The substituents may be halo-, carboxymethyl, guanidino-, guanidinomethyl, carboxamidmethyl, aminomethyl, nitro-, methoxy- or methyl. Examples of these preferred substituents that may be mentioned are phenacetyl, 3-bromophenylacetyl, raminomethylphenylacetyl, 4-carboxymethylphenylacetyl, 4-carboxamidmethylphenylacetyl, 2-furyl-acetyl, 5-nitrofurylacetyl, 3-chloroacetyl, 3-chloroacetyl,

5-methoxythienylacetyl, α-guanidino-

2-thienylacetyl, 3-thienylacetyl, 4-methylthienylacetyl, 3-isothiazolylacetyl, 4-methoxyisothiazolylacetyl, 4isothiazolylacetyl, 3-methylisothiazolylacetyl, 5-isothiazolylacetyl, 3chloroisothiazol, 1,2-azolazolylacetyl

5-Oxadiazolylacetyl, 1,2,5-thiadiazolyl-4-acetyl, 3-methyl-1,2,5-thiadiazolyl-4-acetyl, 3-chloro-1,2,5-thiadiazolyl-4-acetyl, 3- methoxy-1,2,5-thiadiazolyl-4-acetyl, phenylthioacetyl,

Pyrrolidine, pyrrolidone, pearl, pearl, pearl. hydroxyphenylacetyl, α-tetrazolyl-phenylacetyl and α-sulfophenylacetyl.

The substituent A in formula I may be hydrogen, hydroxy, halo, mercapto, acyloxy, acylthio, substituted hydroxy, substituted mercapto, quaternary ammonium, azido, amino or N-substituted amino. Optionally, CH ^ A may be substituted by a formyl group. Thus, CH ₂ A can be halomethyl such as chloromethyl, bromomethyl or fluoromethyl.

When A is a substituted hydroxy or substituted mercapto group, it may be represented by the formula

- ch ₂ zr ₅ in which Z is oxygen or sulfur and R ₅ is an acyl group; straight or branched chain lower alkyl, alkenyl or alkynyl; an aryl group; aralkyl stupid; a heterocyclic group such as heteroaryl or heteroalkyl. These groups may be unsubstituted or substituted by radicals such as alkyl, alkoxy, halo, cyano, carboxycarbamoyl, azido, sulfo, amino, substituted amino, haloalkyl, carboxyalkyl, carbamoylalkyl, N-substitutedylatedyl N-substituted guanidino-, guanidoalkyl, sulfamide, substituted sulfamide and the like. Representatives of said groups to be mentioned are methoxymethyl, n-propoxymethyl, methylthiomethyl, acetoxymethyl, propionyloxymethyl, benzoyloxymethyl, (p-chlorobenzoyl) oxymethyl, (p-methylbenzoyl) oxymethyl, pivaloyloxymethyl, pivaloyloxymethyl, pivaloyloxymethyl, pyaloyloxymethyl, , carbamoyloxymethyl, (N-methylcarbamoyl) oxymethyl, (N-ethylcarbamoyl) oxymethyl, (N- (2-chloroethyl) carbamoyl) oxymethyl, (N-phenylcarbamoyl) oxymethyl, (N-p-sulfophenylcarbamoyl) carbamoylmethyl) oxymethyl, methoxycarbonyloxymethyl, isobut anoiloksimetil, tsiklobutilkarboniloksimetil, karbamoiltiometil, / etoksitiokarbonil / thiomethyl, / n-propoksitiokarbonil / thiomethyl, / tsiklopentanoksitiokarbonil / thiomethyl, methylthiomethyl, Ν, Ndietiltiokarbamoiltiometil, N-methylpiperazin-1 -tiokarboniltiometil, N, N-dimethylpiperazine-1-tio5 karboniltiometil, 2- furoylthiomethyl, isothiouroniummethyl, (5-methyl-1,3, 4,4-thiadazol-2-yl) thiomethyl, p-tolylsulfonylthiomethyl, mesyloxymethyl, 1methyl-1,2,3,4-tetrazolyl-5-thiomethyl, tosyloxymethyl-sulfonylmethylsulfonylmethylthiomethylsulfonylmethylsulfonylmethyl , 2-furylacetoxymethyl, cinnamyloxime methyl, p-hidroksitsinamoiloksimetil, p-sulfotsinamoiloksimetil and 1R: 2S -epoksipropilfosfoniloksimetil.

In a case where CH ₂ A is hydroxymethyl, cephalosporin may also exist as lactone, which is formed by internal esterification with the carboxyl group.

The CH ₂ A substituent may also be a group of the general formula

- CH ₂ Y in which Y is amino or substituted amino, including nitrogen, heterocycles and substituted heterocyclic groups. The groups that may be mentioned are aminomethyl, acetamidomethyl, carbamoylaminomethyl, N, N-dimethylaminomethyl, N- (2-chloroethyl) aminomethyl, 5-cyanotriazol-1-ylmethyl, 4-methoxycarbonyltriazol-1-ylmethyl.

If A is an amino-, the cephalosporin compound may also exist as a lactam formed by cleavage of water in a neighboring carboxyl group.

Representatives of the quaternary ammonium groups designated by the symbol A that may be mentioned are pyridinium, 3-methylpyridinium, 4-methylpyridinium, 3-chloropyridinium, 3-bromopyridinium, 3-iodopyridinium, 4-carbamoylpyridinium, 4- (pyridylmethyl) ,

4- (N-carbomethoxycarbamoyl) pyridinium, 4- (M-cyanocarbamoyl) pyridinium, 4- (carboxymethyl) pyridinium, 4 (hydroxymethyl) pyridimium, 4- (trifluoromethyl) pyridinium, quinoline, picolinium and lutidine.

The preferred groups designated by the symbol A are hydrogen, halo-, azido-, cyano-, hydroxy-, alkoxy-, aryloxy-, aralkyloxy-, heterocycloxy, mercapto-, alkanoylamino-, hydroxyphenyl, acylthio-, acyloxy-, isothiounia, isothioux sulfamoyloxy, quaternary ammonium, heterocyclic tertiary amine, alkylsulfonyloxy and / cis-1,2epoxypropyl / phosphono. Heterocycles may be 5 or 6 membered heteronuclei containing one or more nitrogen, oxygen or sulfur atoms. The acyl group may be a lower alkanoyl group of 2 to 6 carbon atoms, carbamoyl or thiocarbamoyl and N-alkyl or N, N-dialkyl derivatives thereof. The alkyl group of the aforementioned substituents contains from 1 to 6 carbon atoms, and radicals such as alkoxy, halo, amino, cyano, carboxy, cullo and the like may be substituted.

The substituent R1 in formula I may be a hydroxy, mercapto or substituted hydroxy and mercapto group; a hydrocarbyl or substituted hydrocarbyl group; a cyano or carbonyl or thiocarbonyl radical; a nitrogen-linked group; halo- or phosphono- or substituted phosphono group.

The oxy- or thio-substituents denoted by R 1 in formula I may be a hydroxy or mercapto or substituted hydroxy or mercapto group such as R wherein X is oxygen or sulfur and R 'is a hydrocarbon group, preferably a lower alkyl group, straight or branched chain having from 1 to 6 carbon atoms, straight or branched chain lower alkenyl or lower alkynyl group having from 3 to 6 carbon atoms, a single ring aryl group such as phenyls aralkyl group such as benzyl. These alkyl, alkenyl, alkynyl, aryl or aralkyl groups may be substituted by groups such as hydroxy, halo, nitro, amino, carboxy, sulfo and the like. Other specific substituents represented by the symbol R that may be mentioned are the groups of the formulas -OCN, -SCN, -ONR ₃ R ₄ , -OAc, -SAc, -co ₃ h, -co ₃ r ₂ , -co ₂ nh ₂ , -ax ₃ , -co ₂ nr ₃ r ₄ , -co ₂ r ₂ , -ocoor ₂ , -soh ₂ , -O-COSR ₂ , -OCONR ₃ R ₄ and the like, where Ac represents an acyl group such as lower alkanoyl, R ₃ and R ₄ represent hydrogen, lower alkyl, acyl and lower alkoxy and R ₂ represents lower alkyl, halogen alkyl, aryl, aralkyl and substituted derivatives of such groups.

When R1 is a hydrocarbon or substituted hydrocarbon, it may be lower alkyl, lower alkenyl, lower alkynyl, aralkyl, cycloalkyl, a single-ring aryl group, or a one-ring heterocyclic group, which may also be substituted by one or more hydroxy groups, such as halo, -, alkoxy-, amino-, nitro-, sulfonyl, sulfamoyl, acyloxy, carboyloxy, carboxy, carboxamido and N-substituted carboxamido.

R1 in formula I represents a cyano or group of the general formula - CX'R in which X is oxygen or sulfur and R is hydrogen, halo, hydroxy, mercapto, amino, substituted amino, aliphatic radical, aromatic radical , aliphatic oxyadical or aromatic oxyadical. Examples of substituents that may be mentioned are -COOH, -CSSH, -COR ₂ , -coor ₂ , -cosr ₂ , -cssr ₂ , -conh ₂ , -csnh ₂ , -csr ₂ , -conhr ₂ , -csnh , -CONR ₃ R ₄ and -CSNR ₃ R ₄ in which R ₂ denotes a straight- or branched-chain alkyl group having 1 to 6 carbon atoms and R ₃ and R ₄ denote hydrogen or R ₂ .

R ₍ in formula I is linked to a nitrogen group such as amino and substituted amino groups, nitro-, azido-, nitroso-, isocyanate and isothiocyanate and hydroxyamino groups. Examples of nitrogen-related groups to be mentioned are -NH ₂ , -NHR ₂ , -NHC (O) _n R ₂ , NHC (S) _n R ₂ , -NR ₂ R ₃ , -NHNHj, -nhnr ₂ r ₃ ,

C (O) _n RC (O) _n R

-NHN ζ, -NHN

-nnr ₂ , -nr ₃ oh, -nhcnhnh ₂ , -nhcnhnr ₂ r ₃ , -no ₂ , -no, -nco, N ₃ and -NCS, in which R ₂ denotes a lower alkyl group of 1 to 6 carbon atoms, R ₃ is R ₂ or hydrogen and η is an integer of 1 or 2.

The substituent Rj in formula I means a phosphono or a metal or amine salt of the same, or a substituted phosphono group of formula

Oh

T y ~ ^p \

Z 'in which Y' and Z 'are the same or different and denote -OR ₂ , -NR ₃ R ₄ ,

R

I

-NR-CH-COOH, -NR-NR ₃ R ₄ ,

NR,

II ²

-nr ₂ n = cr ₃ r ₄ , -nr ₂ -c-nr ₃ r ₄

X 'X'

II II

-NH-C-X'R ₂ , -nh-c-nr ₃ n ₄ ,

-NC = X ', -OCOR ₂ and -N ₃ where R ₂ denotes hydrogen or hydrocarbyl, R ₃ and R ₄ denote hydrogen, hydrocarbyl, alkoxy or acyl radical and X' denotes oxygen or sulfur. 5

In accordance with the nomenclature used for cephalosporin compounds, the compound obtained by hydrolysis of cephalosporin C, which has the following structural formula

CH2OCOCH, is called 7-aminocephalosporanic acid or 7-ACA. 15

The term "decalphalosporanic acid" used to describe certain products according to their use in this field means a basic heterocyclic nucleus of structural formula

So a compound of formula

is called 3-methyl-7-aminodecephalo-35 sporic acid using the nomenclature system described.

The cephalosporin compounds of the invention are also referred to as "cepham" compounds containing the basic fused ring β-lactam thiazine structure

which is known as cefam. Cephalos- "cephem", when the basic structure of the spiron compounds is called, has a simple olefinic bond. With this one

C having the structural formula nomenclature system cephalosporin

UNSD about n / II

CH (CH ₂ ), - C - N- ^ NH, o '

is called 7- (5'-aminoadipamido) -3acetoxymethyl-3-cepham-4-carboxylic acid.

According to the invention, it is intended to be COOH, but that new cephalosporins can be prepared by a method that can be described as follows

in which R ', R, and A are as defined above.

In the above scheme, the starting compound is a derivative of 7-aminocephalosporanic acid (II), hereinafter also called 7-ACA, where the carboxy group is preferably blocked, for example by forming a suitable ester. In such a way

7-ACAs or analogs thereof with different substituent at the 3-position can be esterified according to methods well known in the art to obtain, for example, esters in which R _g represents an alkyl or substituted alkyl group such as methyl, t-butyl , pivaloyloxymethyl, acetoxymethyl and the like, haloalkyl 45 such as trichloroethyl, alkenyl group such as allyl, alkynyl group such as propargyl, aralkyl group such as benzyl, benzhydryl, o-nitrobenzyl, 3,5-dinitrobenzyl or p-methoxybenzyl or p-methoxybenzyl group , e.g. Air silyl group such as trimethylsilyl or stanilna group such as tributyltin, phenacyl or trichloroethoxycarbonyl. The ester (II) is converted to the corresponding 7-diazocephalosporanic acid ester or the 3-CMj A-7-diazocephalosporanic acid ester (III) by reaction with nitrite. The 7-diazoester (III) is converted by reaction with a pseudo-halogen compound or with a compound that acts as a pseudo-halogen to give an intermediate (IV) in which X represents a halogen of the bromine, chlorine and iodine group or other leaving group, and

Y is a nitrogen substituent or R _r The intermediate (IV) is then converted to the compound (I) in which R ₍ represents a substituent other than hydrogen and Z represents a nitrogen-containing group which can easily be converted to an amine or acylamine. (V) is then converted to the desired cephalosporin ester (VI), which can be reacted to give the corresponding cephalosporic acid or a salt thereof. Also, the substituent at position 3 of the A ^3- cefam nucleus can be converted to the other substituents of the formula -CH ₂ A in accordance with zvestnite in the art and by methods described herein. The processes that take place at any stage of the above scheme, may more readily be understood from the detailed description of the methods that follow.

The starting material for the previous process may be 7-ACA or 3CH ₂ A-7-aminodecifalosporanic acid, which first reacts to block or protect the carboxyl group. Examples of particularly suitable 3-CH ₂ A-7-aminodecephalosporanic acids, which may be mentioned, are those in which A represents hydrogen, hydroxy-, azide-, halo-, tertiary amine, isothuronium, lower alkoxy or lower alkylthio, an acyloxy or acylthio group, or a heterocyclic oxy or heterocyclic thio substituent. When substituent A is halo, it may be fluorine, chlorine or bromine. When A represents a lower alkoxy or lower alkylthio group, it may be a group such as methoxy, tertiary butyloxy, tertiary butylthio and the like. When A is an acyloxy or acylthio group, it may be an acetoxy-, benzoyloxy-, cinamoyloxy-, p-sulfocinamoyloxy-, isobutyryloxy-, pivaloyloxy-, adamantyloxy-, carbamoyloxy-, n-methylcarbamoyloxy-, N-methyl-carboxy- N-carboxymethylphenylcarbamoyloxy, N-chloroethylcarbamoyloxy-, N, N-diethyldithiocarbamoyloxy-, N, N-dimethylpiperidine dithiocarbamoyloxy, mesyloxy-, sulfamoyloxy- and 1R: 2S -1,2-epoxypropyl. If A is a heterocyclic oxy- or heterocyclic thio group, it may be a group such as 5-methyl-1,3,4-thiadiazolyl-2-thio and 4-carboxamido-1,3,4-thiadiazolyl-2-thio. When A is a tertiary amine, it may be pyridine and the like.

The diazotization of the 7-aminoester is carried out in accordance with methods known to those skilled in the art. Thus, diazoitization is conveniently carried out in an aqueous medium or in an environment of water and an organic solvent, for example by reaction with sodium nitrite in the presence of acid or by reaction with organic nitrite. Organic solvents suitable for carrying out the reaction are those which do not contain active hydrogen. Examples of such solvents are methylene chloride, ether, benzene, toluene, chloroform and the like. The reaction is preferably carried out at temperatures between 0 ° and 50 ° C, usually most conveniently carried out at room temperature. Isolation of the desired diazo compound is readily accomplished by methods known in the art. According to a specific embodiment of the invention, the new cephalosporins are prepared by the following method:

COOR, III

COOR, *

wherein the substituents are as defined above. 25

In the above process, the 7-diazocephalosporanic acid ester (III) is reacted with a haloazide of the bromine, chlorine or iodazide group, preferably in the presence of a tertiary amino amino azide to give the 7-halo-7azidcephalosporanoic acid ester (VII), reacting with a suitable nucleophilic reagent is converted into the 35 desired 7 ^ ₍ -7-azidocephalosporanic acid ester (VIII). This intermediate is reduced and acylated in one step to form the substituted cephalosporan ect-40 ter (XI), from which then you can separate the protecting group to yield cephalosporanic acid or salt thereof / X /. Alternatively, as shown in the diagram, an ester of 45 T ^ -T-azidotsefalosporanovata acid / VIP / reduced to the ester of _7-1 7-aminocephalosporanic acylable acid to give the 7-R-7-acylaminocephalosporanic acid ester (XI) or the ester group of the compound (IX) can be cleaved to give the free acid (X), which may be acylated to give the desired substituted cephalosporin or salt thereof. The cleavage step of the blocking group is easily performed according to methods known to those skilled in the art. For example, an aralkyl group such as benzyl ester is removed by reduction, silyl ester can be removed by hydrolysis to give the free acid or salt thereof, and the benzhydryl group is easily cleaved by reaction with trifluoroacetic acid in the presence of anisole. Other esters that are easily removed from the free acid such as trichloroethyl phthalamidomethyl, succinimidomethyl, rmethoxybenzyl, o-nitrobenzyl, phenacyl and t-butyl and the like can be used in this method. Also, as discussed above, the 3-substituent of the D ^3- cefam nucleus can be varied according to methods known to those skilled in the art to obtain the substituted cephalosporins of formula I.

The step of preparing the haloazide intermediate is accomplished by reacting the diazo compound with haloazide at a temperature between about 0 ° and 50 ° C for sufficient time to effect the formation of the desired compound. Preferably, the reaction is carried out in an environment of a suitable organic solvent that is inert to the reactants. Various solvents which do not contain active hydrogen such as methylene chloride, chloroform, toluene, ether and the like or mixtures thereof are suitable reaction media. Preferably, the reaction is carried out in the presence of a second azide such as lithium azide or tertiary ammonium azide, for example triethylammonium azide, since under these conditions the formation of the unwanted 7-dibromo compound is avoided. Haloazide is used in a slight excess of the stoichiometrically required amount. The amount of the second azide is not critical and it is generally desirable to use in excess to obtain maximum yields of the desired haloazide compound under optimal conditions. Upon completion, the formation of the haloazide product is separated and can then be purified, for example by chromatography, according to methods well known in the art.

The next step of the process, involving the replacement of the halosubstituent with a nucleophilic group, is the interaction of the haloazide with a substance capable of providing a group to replace the halogen. Preferably this reaction is carried out in the presence of a suitable reaction inert solvent such as methylene chloride, chloroform, benzene, toluene, ether, petroleum ether and the like; again it is desirable to avoid the use of any solvents containing active hydrogen. According to a preferred embodiment of the invention, the nucleophilic displacement reagent may be an alcohol such as methanol, ethanol, benzyl alcohol, a substituted alcohol such as 2-bromoethanol, 2-methoxyethanol, glycolamide, a glycolic acid ester, and the resulting displacement methoxy, ethoxy, phenoxy, benzyloxy, 2-bromomethoxy, methoxy, 2methoxyethoxy, carbonylmethoxy, or an esterified carbonylmethoxy substituent respectively. The reaction is preferably carried out in the presence of a heavy metal cation such as a silver salt.

When the reaction is carried out by reaction of an organic acid salt, preferably a heavy metal salt such as a silver salt, the corresponding 7-acyloxy compound is obtained. For example, the reaction of haloazide with silver benzoate, silver tert-butyl acetate, silver phenyl acetate results in the corresponding 7-acetoxy, 7-benzyloxy, 7-tert-butylacetoxy and 7-phenylacetoxy intermediate. The acyl groups of these various acyloxy compounds are then cleaved to give the corresponding 7-hydroxy compound. Optionally, by this process for the preparation of 7-acyl 12-lox compounds, the reaction can be carried out using a salt of the appropriate acid and carried out in the presence of a heavy metal salt, for example silver oxide or silver tetrafluoroborate.

In the next step of the process described above, _7-a3H4-7-R] compound can be reduced to yield the corresponding 7-aMMHO-7R ₁ compound. Various methods may be used to effect this reduction, but it is generally preferred to reduce the azide to the amino group by catalytic hydrogenation using a noble metal catalyst such as platinum, palladium or oxides thereof. These processes are performed according to methods well known to those skilled in the art. Optionally, the reduction may be carried out in the presence of a suitable acylating agent to give the desired 7-acylamino-R 2 compound. The 7-amino compound can be reacted with suitable acylating agents using well known procedures to obtain the desired 7-acylamino compound. Thus, by the method described above, wherein the substituent R is a halo group, e.g.

The 7-azido-7-halo compound can be reduced to the corresponding amino compound, which can then be acylated to give the 7-acylamino-7-halo product. In one case, as discussed above, acylation and the reduction step can be combined to give the 7-acylamino compound without the separation and acylation of the 7-acylamide intermediate.

These 7-amidocephalosporanate products, where the substituent at position 7 of the cefam nucleus is attached to the 7-carbon atom via a nitrogen atom, are conveniently synthesized10 from their corresponding 7-halo-7-azide precursors. According to this method of production, 7-halo-7-azidocephalosporanate is converted to the corresponding 7, 7-diazidocephalosporanate by 15 treatment with an alkali metal azide and this intermediate is then reduced by hydrogenation in the presence of a suitable catalyst, for example palladium on 20 charcoal. The resulting 7-amino-7-azidocephalosporanate is then acylated by treatment with acyl halide, carboxylic acid anhydride or sulfonyl halide, and thus the resulting 7-amido-7-azidocephalosporanate is reduced and then converted to free acid to obtain the desired product. The following Scheme 30, wherein the acylating agent used is acyl halide, illustrates this method of preparation; however, it should be understood that any other acylating agent can be used in an analogous reaction to obtain the desired product 7-amido- or 7-sulfonamidocephalosporanic acid:

7-Amido-7-aminocephalosporanic acids according to the invention are intermediates which react well with the amine nitrogen atom with a wide range of reagents to produce N-substituted and N, N-substituted derivatives thereof. For example, 7-amido-7-aminocephalosporanic acid can be reacted with one or more equivalents of an aldehyde such as formaldehyde, acetaldehyde or propionaldehyde and the like or aralkaldehyde such as benzaldehyde and the like to give the corresponding 7-amido-7-alkyl-7-alkyl -Cephalosporanic acid.

In addition to the reaction with the aldehydes, 7-amido-7aminocephalosporanic acid can be treated with an acylating or sulfonating agent such as acyl halide, carboxylic acid anhydride, alkanesulfonyl halide or pyridine-sulfur trioxide complex, or the 7-amidosaldehyde 7-amidosaldehyde 7 -sulfonamido) cephalospanic acid.

Those 7-amido-7-aminocephalosporanic acids, in which the 7-amino radical is substituted with urea or N, N-dialkyl urea, are conveniently obtained by treatment with a suitable carbamoyl halide or N, N-dialkylcarbamoyl halide. Similarly, the derivatives of 7-amido-7-guanidinephalosporanic acid are obtained by simply treating the parent 7-amido-7-aminocephalosporanic acid with N-amidino-3,5-dimethylpyrazole.

7-Amido (7-amidinoureid) cephalosporanic acid derivatives are prepared by first treating the starting 7-amido-7aminocephalosporanic acid with phosgene to give an intermediate, which after treatment with guanidine yields the desired product.

In one embodiment, according to another embodiment of the invention, the 7-aminocephalosporans are also obtained 15 using the 7-azido-7-halo benzhydryl ester of formula VII as the starting material. This compound is reacted with tert-butylcarbamate to give the corresponding Ί20 tert-butylcarbonylamino compound. Reduction of this halogen intermediate in the presence of platinum oxide yields 7-amino-7-tert-butylcarbamoylaminobenzhydryl ester. The latter compound is then acylated to give benzhydryl-7-acylamide-7-tert-butylcarbamoylamino compound which, upon treatment with trifluoroacetic 30 acid in the presence of anisole, gives 7-aminocephalosporin.

7-Amido-7-phosphonone compounds and

The 7-amido-7-phosphinyl products of the invention and their corresponding derivatives, salt and ester, are prepared by treating the 7-azido-7-halocephalosporate compound with a suitable phosphite, phosphonamidic acid or diamidphosphoric acid in the presence of a metal salt, e.g. such as silver oxide or silver tetrafluoroborate and the like. The 7-azido-7-phosphono (or 7-phosphinyl) compound thus obtained45 is then reduced to the corresponding 7-amino-7-phosphono (or 7-phosphinyl) cephalosporanate and subjected to acylation by treatment with acyl halide, anhydride carboxylic anhydride to give the corresponding 7-amide-7-phosphono (or phosphinyl) compound and this intermediate is then isolated and purified or, if desired, said ester can be converted to the corresponding free acid as described above. Also, after treatment with a base, said acid can be converted to the corresponding 7-amide-7-phosphono (or phosphonyl) salt.

When the haloazide compound is reacted with carbon dioxide or carbon disulfide in the presence of phenyllithium, the corresponding 7-azido-7-carboxy or 7-azido-7-thiocarboxy compound is obtained. These carboxy or thiocarboxy compounds can be converted to the corresponding haloformyl compound by reaction with halogenating agents in a manner well known in the art. For example, the 7-carboxy-7-azido compound by reaction with thionyl chloride is converted to a 7-chloroformyl7-azido compound which can be reduced to a 7-amino-7-chloroformyl compound and acylated to give the desired cephalosporic acid or decalphalosporin. Further, the 7-haloformyl compound by reaction with alcohol 5 such as methanol, phenol or benzyl alcohol is converted to the corresponding 7-methoxycarbonyl, 7-phenoxycarbonyl or 7-benzyloxycarbonyl. After the reaction of the 7-haloformyl compound with an amine such as dimethylamine, dibenzamine, diphenylamine, monoethylamine, monobenzylamine, monophenylamine, phenethylamine, hydrazine or substituted hydrazine is converted to the corresponding 7-carboxamide. The compounds of 7-carboxycepharosporanoic and decalphalosporanic acid are also prepared by oxidizing the corresponding 7-formyl compounds 20 with silver oxide. The 7-formyl compounds are prepared by treating the 7-hydroxymethyl-substituted product with phosphoric acid at pH 2-3 to give the 7-hydroxy 25 compound and then the latter products are oxidized with a pyridine-chromium trioxide complex.

The new cephalosporanic and decalphalosporanic acids, where R 1 is a 30 hydrocarbon group, are obtained by the reactions shown below:

dream ₂ a

dream ₂ a

wherein D is a hydrocarbyl group and R ₍ and A are as defined above.

According to the above scheme, the diazocephalosporin compound is reacted with the trihydrocarbylborne compound at low temperatures, for example from -50 ° to -100 ° C for a sufficient time to obtain the intermediate

7-Dihydrocarbylbor-7-hydrocarbon (XVII). Reaction of this intermediate with halogenazide as bromazide at room temperature gave 7-hydrocarbyl-7-azido compound (XVIII). The latter compound is then catalytically reduced, acylated, and the ester group cleaved according to the methods described above to give the desired 7-hydrocarbyl-7-acylamidoce phalosporanic or decephalosporanic acid (XIX) or a salt thereof.

In the first step of this procedure, the hydrocarbyl group at the boronic 5 compound may be a lower alkyl group of 1 to 6 carbon atoms, a lower alkenyl group of 2 to 6 carbon atoms, a lower alkynyl group of 2 to 6 carbon atoms, an aralkyl 10 group such as benzyl or an aryl group such as phenyl. Using these trisubstituted boron compounds, the corresponding arylcephalosporanic acid is obtained.

According to one embodiment of the invention, new cephalosporins with 7carboxy or substituted carboxy substituents are prepared as follows:

snl

nooo

COOR,

XXI

According to one of the above methods, the resulting intermediate (IV) as described above is reacted with hydrocarbillithio of formula R ₁₀ Li, where R ₁₀ represents a hydrocarbonyl group such as lower alkyl or aryl, for example n-butyllithium, to form 7 -Lithium compound (XX), which reacts with carbon dioxide to give

7-α-carboxy compound (XXI). This intermediate is converted to carboxy 7-cephalosporin (XXII) using the methods mentioned above or the carboxy substituent can be converted to a carboxylic acid derivative such as ester, amide, hydrazide, azide or hydroxamic acid using known procedures. In a case where

The 7-lithium compound was reacted with carbon disulfide instead of carbon dioxide to give the corresponding 7-dithiocarboxy (-CSSH) compound. 5

The 7-cyanocephalosporins are prepared as the intermediate 7-halo-7-azido compound of formula VII above, reacted with tetrabutylammonium cyanide to give the 7-cyano-7-azide compound. This intermediate was then reduced to the 7-cyano-7-amino compound, which was acylated and the ester removed to give 7-cyano-7-acylamidocephalosporin using the procedures described above.

The 7-formylcephalosporins are prepared by converting the 7-hydroxymethyl 1-7 -cylamide ido-cephalosporic acid or the corresponding 3-CH ₂ A decephalosporanic acid with an oxidizing agent such as pyridine chromium trioxide to give the 7-formyl compound. This latter cephalosporin compound is converted to the corresponding 7-carboxyproduct by soft oxidizing agents such as silver oxide.

The 7-halo-substituted cephalosporins of the invention are prepared by reducing the 7-halo-7-azidamide compound of formula VII to give the corresponding 7-halo-7-amine compound, and this intermediate is acylated to the corresponding 7-acylamino-7-halocephalose. The resulting ester 15 is then cleaved and converted to its corresponding carboximate salt in known ways, for example by treatment with trifluoroacetic acid and an aqueous base solution.

In another embodiment, the new 7-hydrocarbonyloxy and 7-hydrocarbylthiocephalosporins can be prepared by the following scheme:

CH ₂ a where R 1 and A have the meanings defined above and G represents hydrocarbonyloxy or hydrocarbylthio.

According to the above sequence, the starting compound, an ester of 7-diazo compound, designated III, is reacted with a hypocholite in alcohol or thiol or with alcohol in the presence of a positive halogen, such as N-haloamide, for example N bromoacetamide, N-bromosuccinimide, 45 N-bromophthalimide and others which react as the corresponding hypochalite. The resulting 7-halo-7-hydrocarbonyloxy or hydrocarbylthioester (XXIII) is often a mixture of 7-site epimers that can be separated by chromatography. When only one epimer is obtained, it can be equilibrated as a mixture of eipermers by treatment with inorganic halide in a polar solvent. The lithium salt of the appropriate halide in dimethylformamide is usually used to epimerize these intermediates. The 7-halo-7-hydrocarbonyloxy or hydrocarbylthio product can then be reacted with an azide such as lithium azide to form 7-hydrocarbonyloxy or hydrocarbylthio-7-azidocephalosporanate ester (XXIV). The latter compound can then be reduced either with hydrogen or with an inorganic reducing agent to give the intermediate 7-hydrocarbonyloxy or hydrocarbylthio-7-aminoester (XXU R-H).

The latter compound may be acylated to give the substituted cephalosporin ester. In another case, the reduction of the intermediate azido compound may be made in the presence of an acylating agent to obtain these esters directly. These compounds can then be converted to the desired cephalosporin of formula XXV or a salt thereof according to the procedures described above.

The various methods described above may result in polu5's appointment partial epimer at 7th position or a mixture of epimers at the 7th place, for example, 7-halo-73-R ₁ or 7.beta.-halo-7-AND ₁ circuit. When a mixture of epimers is obtained, they can be easily separated by methods, for example, chromatographically well known to those skilled in the art. In some cases, when only one epimer is obtained, it can be equilibrated to produce a mixture of epimers according to methods known to those skilled in the art.

According to another embodiment of the invention, the novel products are also obtained by a novel method in which the acyl group at 7-acylamidocephalosporin is replaced by a different acyl substituent. In accordance with this new method, 7-acylamidocephalosporin is reacted with an acylating agent to obtain the intermediate 7-diacylamidocephalosporin containing two different acyl substituents and then the original acyl group is cleaved 30 to give the new 7-acylamidocephalosporin. This process is illustrated by the following scheme:

AcHN

dream ₂ a

where A represents an acyl group A ', R / and R _p represent respectively the substituents defined as A, R _p and R', or may be converted back into them by removing any protecting or blocking group.

In the process described in the above scheme, the reaction can be carried out with the free acid, although it is generally found that it is preferable to block or protect the carboxyl group by forming a suitable ester which can then easily be removed at the end of the process.

The first step of this process involves the interaction of cephalosporin or a derivative thereof, wherein the carboxyl group is blocked with an acylating agent, preferably an acyl halide, in the presence of a silyl group to give the 7-diacylamide compound. The original acyl substituent was then removed from the product to give the cephalosporin compound with the new 7-acylamide substituent.

The first step of preparing the diacylated product is best accomplished by intimate contact of the cephalosporin with the acylating agent in a suitable solvent medium in the presence of a three-substituted silyl derivative of an electronegatively substituted amide. The temperature at which the reaction is carried out is not particularly critical and temperatures from about -20 ° C to about 100 ° C are most appropriate, although temperatures from about 25 ° C to 40 ° C are preferred. Various solvents that do not contain active hydrogen such as chloroform, acetonitrile, methylene chloride, dioxane, benzene, halobenzene, tetrachloromethane and diethyl ether are most suitable as reaction media.

Various trihydrocarbylsilyl compounds wherein the hydrocarbyl substituent is lower alkyl (from 1 to 6 carbon atoms), aryl such as phenyl or an aralkyl group such as benzyl, may be used in the process of the invention. These compounds are readily prepared by reacting equimolecular amounts of trihydrocarbylsilyl halide with a negatively substituted amide or imide. It is generally preferred to use a tri-lower alkylsilyl derivative, and in particular a trimethylsilyl derivative, since this product is inexpensive and readily available. The negatively substituted amides and imides that should be mentioned are succinamide, phthalimide, cyanoacetamide, trifluoroacetamide, benzamide, p-nitrobenzamide, trichloroacetamide, sulfonamide and the like. Particularly useful trisalkylsilyl derivatives are N-trimethylsilyltrifluoroacetamide, N-trimethylsilylphthalamide.

The above reactions are preferably carried out with a cephalosporin compound in which the carboxy group is blocked or protected, since maximum yields of the desired product are obtained with such derivatives. For this purpose, the carboxy substituent is blocked by the formation of a suitable ester such as benzyl, benzhydryl, p-nitrophenyl, trimethylsilyl, trichloroethoxy, p-methoxybenzyl, phthalimidomethyl or succinimidomethyl ester, which are easily removed by methods well known in the art. It is generally preferred to block or protect any available amino group in the starting cephalosporin compound, since such derivatives produce maximum yields of the desired products. Preferably, the groups are blocked with substituents which can be eliminated 19. Such groups are well known to those skilled in the art. For example, the amino group is conveniently blocked with a group such as trichloroethoxycarbonyl, tert-butoxycarbonyl, benzoylmethoxycarbonyl, trimethylsilyl, p-methoxybenzyloxy, o-nitrophenylthio and the like.

The cleavage step of the initial acyl group can be accomplished in several ways, by continuing the reaction time by adding an alcohol such as lower alkanol or lower alkylthiol or by hydrolysis in an aqueous solution containing a small amount of acid or base. In some cases the cleavage is effected by the addition of lower alkanol or lower alkylthiol containing from 1 to 6 carbon atoms, aralkanol such as benzyl alcohol or the corresponding thiol. Cleavage yields the desired monoacylated cephalosporin compound or may also result in a mixture of monoacyl compounds. In the latter case, the desired monoacyl cephalosporin compound is obtained by separation methods, for example chromatography, which are well known in the art.

The method according to this embodiment of the invention is particularly suitable for replacing the aminoadipoyl group of the 7- (aminoadipoylamido) side chain of cephalosporins such as those obtained by fermentation and derivatives thereof having other substituents in the 3rd position. Thus, according to a particular embodiment of this method, a cephalosporin compound such as cephalosporin C or 7- (O-5'-amino-5'-carboxyvaleramido) -3-carbamoylmethyl-7-methoxy-3-cephem-4-carboxylic acid, or derivatives thereof, is reacted with an acylating agent in the presence of a trisubstituted silyl radical to give the циdiacylamido derivative which has two different acyl groups. The diacylated product can be selectively cleaved by removing the α-aminoadipoyl group to give the desired other 7-acylamidocephalosporin compound. Although cephalos challenged the new compound can be transesterified according to the method, it has been found that the process is facilitated and maximum yields of the new 7-acylamido compound are obtained under optimum conditions under which the amino and carboxy substituents of the cephalic compounds are blocked and protected in carrying out the process according to the invention. The various blocking or protecting groups mentioned above are suitable for this purpose. For example, by replacing the α-aminoadipoyl side chain of the cephalosporins mentioned above with another acyl group, following the preferred embodiment of the invention, the two carboxy groups in the 4th position and the carboxy group of the aminoadipoyl substituent are blocked and the amino group is properly protected way. The resulting blocked derivative is reacted with the acylating agent, preferably an acid halide such as chloride, in the presence of a trisubstituted silyl derivative of a negatively substituted amide or imide to give the 7-diacylamido derivative. During this acylation reaction, cleavage of the α-aminoadipoyl group occurs to a certain extent, but most of the product is obtained in the form of a diacylated derivative.

When the protecting group of the amino-substituent of the aminoadipoyl moiety, such as trichloroethoxycarbonyl or tert-butoxycarbonyl group is removed by suitable means, selective cleavage of the aminoadipoyl group occurs. This removal of the amino group protecting group obviously results in internal cyclization of the aminoadipoyl group, which secures the group as the α-carboxyl ester of formula.

ROOC ^ n

Tests conducted in connection with the method indicate that this is the mechanism of such cleavage, which explanation should not be binding, as further studies may establish that the product is cleaved and otherwise extracted. An explanation of how the cleavage occurs is given for a better understanding of the present invention.

The cleavage of the protecting groups of amino and carboxyfunctions is carried out in accordance with methods known to those skilled in the art. For example, the trichloroethoxycarbonyl group is removed by reaction with zinc and acetic acid, and the tert-butoxycarbonyl and benzhydryl groups are separated by reaction with trifluoroacetic acid.

According to another aspect of the invention, the new 7-diacylamide compounds prepared according to the method are not only useful as intermediates 15 for the preparation of monoacylated cephalosporins, but are also useful antibacterial products active against various pathogenic microorganisms.

A more complete understanding of the process according to the invention is provided by the illustrated embodiment which follows. Thus 7- (O-5'-amino-5'carboxyvaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid is converted to the corresponding 7- (2-thienylacetamide) compound according to the method in the following sequence reactions:

UNC

CHjOCONH,

COOH o

II

COCH ₂ CC1,

I

NH OH

I II I

HC- (CH ₂ ), C - N

COOH ο

COOH

CHjOCONH,

O II οι NH I HC- (CH ₂ ) ₂ C —N

COOFN ₂ ο

OCH ₂ CC1 ₂

O H II I

CHjOCONHj coosnf ₂ o jy

II dream,

C - OCH ₂ CC1 ₂ I

NH O,

I II '

HC- (CH,) ₂ C - N

I

COOCH ₂ O

CHjOCONH, soosnf ₂

CHjOCONH, ο

CHjOCONH,

COOH coosnf ₂

In this process, the starting compound is acylated by reaction with trichloroethoxycarbonyl chloride to give the N-trichloroethoxycarbonyl derivative which, after alkylation with diphenyldiazomethane, is converted to dibenzhydryl ester. Reaction of the obtained cephalosporin compound with trimethylsilyltrifluoroacetamide and 2-thienylacetyl chloride yields 7- (D-5-trichloroethoxycarbonylamino-5'-carboxyvaleryl) - (2-thienylacetylamino) compound. This aminoadipoyl group is then cleaved by reaction with zinc in the presence of acid, yielding the 3-carbamoyloxymethyl-7- (2-thienylacetamido) decahalosporic acid benzhydryl ester, which is further unblocked to remove the benzhydryl group. received the free acid. This product can be converted to salt according to methods known to those skilled in the art.

Other acylating agents, such as those defined in R above, may be used in place of the 2-thienylacetyl chloride shown in the scheme to obtain the corresponding 7-acylamidocephem compound. Using these acylating agents, it is necessary to avoid the use of acylating agents containing substituents that may be affected during the reaction. Thus, the amine, carboxy or hydroxy substituents of the acylating agent must be blocked or protected by groups such as those mentioned above and subsequently sequentially stranded. Other specific acylating agents that may be mentioned are phenylacetyl chloride, 2furylacetyl chloride, thiophenoxyacetylchloride, α-azidophenylacetyl chloride and the like. In another case, other acylating agents such as anhydrides or mixed anhydrides may be used in place of acid halides. This 15 transacylation method is a valuable contribution to the art since it provides a means of producing cephalosporins containing different 7-acylamide substituents instead of amino 20-amylamogroup, thus avoiding the need to first convert the known cephalosporin to the corresponding compound of 7-aminosinophosphinol then aci25 pound this product. In addition to the use of cephalosporins obtained by fermentation as starting materials according to the process, derivatives of cep30 phalosporins containing other substituents in the 3 rd place may be used instead of carbamoyloxymethyl or the acetoxymethyl substituent as 3 substituents of the general CH ₂ A formula more than 35 re. In another case, other 3-substituted cephalosporins may be obtained, for example from 3-acetoxymethyl7-acylamidocephalosporins, according to methods well known in the art.

Examples of other cephalosporins having a 7-methoxy or 7-hydrogen substituent, which can be prepared by the method described and which can be mentioned, are shown in the following table

7-acylamide substituent

3-substituent ch ₂ conh - ^ 1ch ₂ conh - φ — S — CH ₂ —CONH—

F

ch ₂ conh—

-CH,

-CH, ίθ

CH ₃ —ch ₂ ocon ^

H - ch ₂ oconh ₂

HC — N —ch ₂ conh

CH ₃ (CH ₂ ) ₃ SCH ₂ —CONH—

H ₃ C

ch ₂ conh— _X ^CH 3

-ch ₂ n ^ ch ₃ —CH ₂ OCH ₃

Cl

I

CH ₂ —CH ₂ —CH, OCON '\ TT

An alternative route to the preparation of the 7- _l 1-7 -amino compounds of formula IX mentioned above involves the interaction of the 7-amino compound of formula II above with an aromatic aldehyde to give the imino-addition compound, treatment of this particular reagent compound with a specific reagent giving a 7-R] Schiff base-attachment and then 30 regeneration of the amino moiety. This method is not part of the present invention but is described and claimed in GB 13008 of 16.03.1972.

Specifically, this alternative route 35 can be used to prepare compounds of the following formula:

where A and R _g are as defined above and R 1 is lower alkyl, lower alkoxy, lower alkylthio, lower alkanoyl, lower haloIX

CH3A alkoxy, lower haloalkylthio-, halo-, lower haloalkyl, lower alkanoyloxy, (α-hydroxy (lower alkyl), α-hydroxy23) (lower alkenyl), β-substituted ethylene derivatives, allyl, benzyl, cyano-, carboxy, cyano-, carboxy -, carbonyl alkoxy, sulfo, sulfonyl; lower alkylsulfo-, phospho-, nitro-, carboxy- and dithiocarboxy.

The starting material is a 7-NH ₂ compound of formula II above, which is reacted with an aromatic aldehyde, preferably one having at least one o- or p-eletron-negative substituent selected from the group consisting of nitro, methylsulfonyl, cyano- , carboxyl derivatives and the like. The preferred reagent is p-nitrobenzaldehyde.

The starting material and the aromatic aldehyde are mixed together in approximately equimolar amounts in an inert solvent. Suitable solvents are dioxane, acetonitrile, dimethylformamide, dimethylsulfoxide, benzene, toluene and the like. If desired, the aldehyde may be used in molecular excess. The reaction proceeds readily at a temperature ranging from ambient temperature to reflux temperature of the solvent. As this condensation is an equilibrium reaction and water is one of the products of the reaction, it is removed from active participation in further reactions by any of a number of conventional methods, including by azeotropic distillation, molecular sieves or borate esters. The specific method depends on the exact reaction parameters. The reaction is carried out by evaporation of the solvent. The imine derivative is then captured and used in the next step.

The latter involves the substitution of R, the group at the carbon 10 atom adjacent to the imino nitrogen. This reaction occurs in the presence of an inert solvent such as those listed above and in the additional presence of an organic or inorganic base. It is preferred to use organic bases such as tertiary amines or pyridine. The particular tertiary amine that is preferred is diisopropylethylamine, although all tertiary lower alkylamine can be used. Inorganic bases such as NaH, NaOH, KON, carbonate or bicarbonate salts and the like can also be used. For example, the reaction may be carried out in "soft glass", which contains a sufficiently soluble inorganic base to catalyze the reaction.

The specific reagents used in reaction with the imine compound to obtain the selected R 1 group depend on the desired R 1 group.

The following table is relevant 35 for determining each reagent with respect to the final Rj group.

Reagent R1 lower alkyl sulfate or halide lower alkyl lower alkanoyl halide lower alkanoyl lower alkyl peroxide lower alkoxy halogenated alkyl peroxide lower haloalkoxy lower alkyl disulfide lower alkylthio halonated alkyl disulfide lower haloalkylthio

terts.butilhipohalit or perhalometilhipohalit halonissh alkane lower alkanoilperoksid formaldehyde or lower alkylaldehyde-reacted lower alkyl ketone reastsionno capable ethylenic derivatives allyl halide benzilahid cyanogen bromide nitrozilhalid karbamoilhalid lower alkyl haloformate sulfuryl sulfamoyl lower alkilsulfonilhalid phosphorus oxychloride atsetontsianhidrinnitrat carbon dioxide karbodisulfid halo halonissh alkyl lower alkanoyloxy (α-hydroxy) lower alkyl (α-hydroxy) branched lower alkyl (β-substituted) methyl allyl benzyl tsianonitrozokarbamoil karbonissh alkoxy sulfosulfamoil lower alkylsulfonylamino fosfonitrokarboksiditiokarboksi ,, / α-hydroxy / lower alkyl "is used to mean a group of the formula

H ι

- with - on ι R where R is hydrogen or alkyl having 1 to 6 carbon atoms.

"Reactive lower alkyl ketone" is used to denote a ketone of the formula

Oh

II

R '- C - R where one of R' and R is a halogenated lower alkyl group, the halogen-substituted carbon being adjacent to the carbonyl function; or one of R 'or R is an alkylcarbonyl group. The carbonyl group is adjacent to the carbonyl of the ketone; the other of R 'or R is lower alkyl. Thus, for example, a type of "reactive lower alkyl ketone" is

XO

III

X '- C - C - R 25ι

X where X is halo-, X is halo or hydrogen and X 'is halo, hydrogen or lower alkyl; and R is lower alkyl.

The other type is

O O II II R - C - C - R where R is hydrogen or lower alkyl and

R is lower alkyl, halogen alkyl, lower alkoxy, or lower haloalkoxy. "(Α-hydroxy) branched lower alkyl" means a group of the formula ⁴⁰ X

I

X '- C - X

I ^{R OH} or

R '

I

- C - OH

And c = o

And R where X, X ', X, R' and R are as defined above.

A "reactive ethylene derivative" is used to refer to an ethylene unsaturated compound that is activated by the presence of one or more highly releasing electron groups, for example compounds of formula CH ₂ = CH ₃ in which Y is

-C = 0, - NO ,, - CN, -C-0 -CH ,, CF,

I ² II ^{3 3}

H O and their like are included in them.

The term "/ β-substituted / ethyl" is used to mean the following group - CH ₂ - CH ₂ Y in which Y is the same as mentioned above.

Following the reaction between the imino compound and the reagent to form the new 7-R 'compounds, the imino part is regenerated to amino-.

This regeneration is carried out by aminolysis or hydrazinolysis in the presence of a catalytic amount of acid. Preferably aniline hydrochloride is used, which serves both as a source of amine and as an acid. When using hydrazine derivatives such as phenylhydrazine, 2,4-dinitrophenylhydrazine and the like, acid is added. Other hydrazines or amines may be used. Preferred environments are lower alkanols such as methanol, ethanol and the like. The usual acids or bases may be used. For example, p-toluenesulfonic acid or aniline hydrochloride may be used. The only restriction is that no unwanted hydrolysis or ring damage occurs.

7 _R) -7-aminocephalosporanic acid and derivatives of the 7-R _| -7-aMHHODecephalosporanic acid of formula IX above, thus obtained, can be converted to cephalosporin compounds following the procedures described above.

The step of acylating the 7-amino compounds of formula IX above is accomplished by reacting the amino compound with acyl acid in the presence of an activating agent of the type dicyclohexyl diimide, with acyl anhydride, with acyl halide such as acid chloride or with an activated acid ester such as p- nitrophenyl ester. In the process of reductive acylation, the 7-azido compounds of formula VIII mentioned above, reductive acylation is preferably carried out in the presence of acyl anhydride.

The following examples illustrate the invention without limitation.

Example 1.

A. 7-Diazocephalosporanic acid benzyl ester

A mixture of 10 g of sodium nitrite, 4.5 g of p-toluenesulfonic acid, 7-aminocephalosporanic acid benzyl ester, 300 ml of methylene chloride and 300 ml of water / ice was shaken in a separatory funnel. P-toluenesulfonic acid monohydrate 1.6 g was added in three portions for 20 min. During this time the separating funnel was shaken vigorously. The methylene chloride layer was separated, dried over sodium sulfate and evaporated under reduced pressure to 4-0 ml.

B. 7-Azido-7-bromocephalosporanic acid benzyl ester

To the solution of benzyl ester of

7-Diazocephalosporanic acid in 40 ml of methylene chloride was added 40 ml of nitromethane and the resulting solution was cooled in an ice bath. To this solution was added 80 ml of triethylammonium azide in methylene chloride prepared as described below. To this reaction mixture was then added 40 ml of methylene chloride containing bromoazide prepared as described below with good stirring. After gas evolution ceases, 200 ml, 1 N sodium thiosulphate are added and the mixture is shaken vigorously.-The organic layer is then separated and 200 ml of water are added. Solid sodium bicarbonate was added to the mixture in small portions until the aqueous phase remained pH 7 after stirring. The organic layer was separated and evaporated under reduced pressure, using 4.8 g of crude 7-azido-7-bromocephalosporanic acid benzyl ester as a brown oil. The product was further purified by chromatography on 120 g of silica gel using elution of the product 1: 1 and 1: 3 hexane / benzene. The fractions containing the pure product were combined and evaporated to give 7-azido-7 bromocephalosporanic acid benzyl ester. IR / flow film (s): 4, 7µ (azide), 5.60 μ (β-lactamine carbonyl), 5.75 μ (esters). NMR / CDCI3 / TMS (100 MHz):

2.01 δ / C, 3H, CH, C - O /,

II

3.42 δ (AB, 2H, S - CH ₂ ), 4.68 and 4.71δ (singlets, C-6 hydrogen). Silica gel thin layer chromatography using 1% methanol and chloroform - R _f = 0.70.

The triethylammonium azide solution was prepared by mixing 6.0 g of sodium azide, 20 ml of water and 50 ml of methylene chloride, cooling this reaction mixture to 0 ° C and adding 6 ml of concentrated sulfuric acid. The resulting reaction mixture was allowed to stir for 10 min, the layers were separated and the aqueous layer was washed with a small amount of methylene chloride, which was added to the pre-separated methylene chloride phase. After drying with calcium chloride, the methylene chloride solution was neutralized to pH 7 with triethylamine and the final volume was adjusted to 100 ml with methylene chloride.

The bromoazide solution was prepared by cooling a mixture consisting of 26 g of sodium azide, 80 ml of methylene chloride and 6.4 g of bromine to 0 ° C, adding 20 ml of concentrated hydrochloric acid, the reaction mixture was stirred in an ice bath over 3 h, the organic phase was separated, the aqueous phase was washed with a small amount of methylene chloride, which was added to the separated organic phase and the volume was adjusted to 100 ml with methylene chloride.

C. 7-Azido-7-methoxycephalosporanic acid benzyl ester

A solution of 7-azido-7-bromocephalosporanic acid benzene ester in methanol is triturated with one equivalent of dry silver fluoroborate. A dark yellow precipitate quickly forms. The mixture was stirred at 22 ° C for 2 and 3/4 hours. The solid was separated by filtration and the filtrate was evaporated to give crude benzyl ester of 7-azido-7-methoxycephalosporic acid. The crude product was chromatographed on silica gel using 2% chloroform in benzene. The desired product has an NMR (CDCI ₃ ™ ^C (partial, 100 MHz), 2.02 δ (S, 3H, CH ₂ C-O),

3.60 δ / C, 3H, OCH ₃ /, IR / flow. film / strong absorption at 4.7µ / azide /, 5.60 μ / β-lactam /, 6.76 μ / esters /. Thin layer chromatography on silica gel C with 1% methanol in chloroform R _f = 0.65.

D. 7-acetamido-7-methoxycephalosporanic acid benzyl ester

After 70.5 mg of 7-azido-7-methoxycephalosporanic acid benzyl ester, 69.5 mg of platinum oxide and 5.0 ml of acetic anhydride were hydrogenated at atmospheric pressure for 16 hours. The solvent was evaporated under reduced pressure and the crude The product was chromatographed on 12 g of silica gel using chloroform and chloroform with 1-5% methanol. The 7acetamido-7-methoxycephalosporanic acid crude benzyl ester was eluted with 1% methanol in chloroform. The product has IR (liquid film / 3.0 μ / NH), 5.60 μ / β lactamine carbonyl /, 5.75 μ / esters /, 5.93 and 6.60 μ / amide / and no absorption at 4.7 μ / azide /. NMR CDCI ₃ ™ ^S (partial, 100 MHz):

2.016 (S, CH ₃ C-O), 2.07 and 2.10 6

O / singlets, CH, C-NH, 3,506 / singlet, O

OCH ₃ /, 5,206 (singlet, C-6 hydrogen). TLC R _f = 0.40 on silica gel G with 5% methanol in chloroform.

E. 7-Acetamido-7methoxycephalosporanic acid potassium salt

7acetamido-7-methoxycephalosporic acid benzyl ester solution (25mg) in 3 ml of 1: 1 aqueous methanol was hydrogenated using a 25 mg 10% Pd / C catalyst at

40.6,895 kN / m ² of hydrogen for 1h. The mixture was filtered and the pH of the filtrate was adjusted to 8 with potassium bicarbonate. The aqueous solution was lyophilized to give the potassium salt of 7-acetamido-7-methoxy cephalosporic acid.

Example 2.

A. Aminocephalosporanic acid benzhydryl ester

To a suspension of 6.8 g / to 0.025 mol / 7-aminocephalosporanic acid in 300 ml of peroxide-free dioxane at room temperature was added 4.3 g / 0.022 mol / p-toluenesulfonic acid monohydrate at stirring. The clear solution was concentrated in vacuo and washed twice with dioxane. The residue was dissolved in 300 ml of dioxane at room temperature and a solution of 10g / 0.05 mol / diphenyldiazomethane in 25 ml of dioxane was added dropwise over 15 minutes. The wine-colored solution was stirred for an additional 30 min, then 25 ml of methanol was added to neutralize the excess C ₂ CN ₂ . The mixture was concentrated in vacuo and the residue partitioned between 200 cc. CH ₂ Cl ₂ and 200 ml of water containing 10 g of K ₂ HPO ₄ (pH 8.5). The organic phase was washed with water, dried over Na ₂ SO ₄ and concentrated in vacuo to an oil. The oil was stirred with 100 ml of ether for 1 h. The precipitate was filtered off, washed with ether and dried to a constant weight of 4.7 g (43%), mp 126-128 ° C. Analysis: Calculated: C, 63.0; H, 5.01; N, 6.37; Found: C, 62.7; H, 5.18; N, 5.18. IR in CHCl _{3 3} is

5.5 μ / β-lactam C = 0 / and 5.8μ / ester C = 0 /. The NMR in SDS1 ₃ is 1.856 (singlet, ΝΗ. /; 2.0 6 (singlet, CH ₃ C); 3.45 6o (doublet, CH ₂ C); 4.86 (singlet, CH ₂ OAc); 4.7 6 / doublet, C ₆ H; 4.9 6 doublets, C H /; 6.986 (doublet, 1 ^H CH);

Φ ^ζ and 7.4 6 (singlet, phenyl).

B. 7-Diazocephalosporanic acid benzhydryl ester

To a stirred mixture of 1.6 g

NaNO ₂ , 30 ml water and 40 ml CH ₂ Cl ₂ at 0 ° C was added 880 mg / 0.002 mol / ester, then a solution of 760 mg / 0.004 mol / p-toluenesulfonic acid in 5 ml water for several minutes. was stirred at 0 ° C for 20 min, then the organic phase was separated, washed with 1x10 ml water, dried over sodium sulfate at 0 ° C, filtered and concentrated in vacuo at room temperature, giving 900 mg of glassy mass. IR: 4.8μ / strong Ν = Ν /,

5.6 μ / β-lactams C = 0 / and 5.8 μ / ester C - 0 /, the NMR in SDS1 ₃ is 2.0 δ / singlet, CH ₃ C = 0 /; 3.4 δ (doublet, CH ₂ S);

4.8 δ (singlet, CH ₂ OAc); 5.6 δ (singlet, C ₆ H); 6.98 δ (singlet, CHCl 1f and 7.4 δ (singlet, phenyl).

C. 7-Bromo-7-azidocephalosporanic acid benzhydryl ester

To a solution of 900 mg of 7-diazocephalosporanic acid benzhydryl ester in 20 ml of CH ₂ Cl ₂ and 10 ml of CH ₃ NO ₂ at 0 ° C was added at once all of Et ₃ N + NH ₃ , then a solution of BrN ₃ , then which was added 50 ml of water, then solid sodium bicarbonate to pH 8.

The organic layer was separated and extracted with 2x20 ml of water, dried over sodium sulfate and concentrated in vacuo to give 900mg (83%).

The NMR spectrum corresponds to the structure. Chromatography on silica gel with chloroform showed a major spot at R ₍ 0.2. Chromatography on 900 mg of crude product on 25 g of chloroform silica gave 400 mg / 39% / material giving one spot as an oil. IR in chloroform: 4 , 72 μ / Ν ₃ /, 5.56 μ / β-lactate C = 0 / and 5.75 μ / ester C = 0 /.

NMR in deuterochloroform: 2.0 δ (synOH, CH ₃ C); 3.38 δ (CH ₂ S); 4, Ί δ (singlet, C ₆ NO); 4.94 δ (CH-O), 6.95 δ (singlet, ^ ^ CH); and 7.48 δ (singlet, phenyl).

Preparation of BrN3 solution

2.66 g / 0.04 mol / sodium azide was added to 8 ml of CH ₂ Cl ₂ at 0 ° C, followed by 0.65 g / 0.0042 mol / bromine. To this stirred mixture at 0 ° C was added dropwise 2 ml of concentrated hydrochloric acid. The mixture was stirred at 0 ° C for three hours.

The organic layer was decanted and the aqueous layer extracted with 1 x 5 ml CH ₂ Cl ₂ . The combined organic phase was stored at -10 ° C.

Preparation of Et ₃ N _H + N ₃ - solution

To a suspension of 1.5 g of sodium azide in 5 ml of water and 10 ml of CH ₂ Cl ₂ at -10 ° C to 0 ° C was added dropwise 4 ml of 50% sulfuric acid. The organic phase was poured into an aqueous paste and the aqueous extract was washed with 1x5 ml of CH ₂ Cl ₂ . The combined organic phase was dried over SaS1 _2. The decanted HN ₃ solution was adjusted to pH 7 with Et ₃ N and stored at -10 ° C.

D. 7-Methoxy-7-azidocephalosporanoic acid benzhydryl ester

To a solution of 400 mg / 0.00072 mol / bromazide in 30 ml methanol was added 150 mg / 0.0008 ml / AgBF ₄ . The mixture was stirred in the dark for 2 1/2 hours. The mixture was concentrated in vacuo and the residue mixed with 50 ml of CH ₂ Cl ₂ was filtered. The filtrate was extracted twice with saturated sodium bicarbonate solution twice with water, dried over anhydrous magnesium sulfate and concentrated in vacuo to give 300 mg / 83% / crystals. Melting point 145-148 ° C. IR in CHCl _{3 3} is 4.72 μ / Ν ₃ lane /, 5.6 μ / β-lactam / and 5.7 μ / ester C = 0 /, NMR is 2.0 δ (singlet, CH, C); 3.4 δ / CH.S/; 3.6 δ / sin3 II ί gl, OCH ₃ /; 4.88 δ (singlet, C ₆ H); 4.95 (CH ₂ O); 6.98 δ (singlet, ^>CH); and 7.4δ (singlet, phenyl).

Analysis: calculated C, 58.4; H, 4.45; N, 11.3; S 6.5; found C, 58.56; H 5 4.65; N, 11.30; S, 5.70.

Example 3.

A. 7-Methoxy-7-aminocephalosporanoic acid benzhydryl ester

1.0 g of 7-azi-10 to-7-methoxycephalosporanic acid benzhydryl ester was dissolved in 100 ml of dioxane. 1.0 g of platinum oxide is added and the reaction mixture is stirred under nitrogen at atmospheric pressure for 1 h. Another amount of 1.0 g of platinum oxide is added and the reaction mixture is again placed under hydrogen and stirred for 3 h. hours until the azide interacts completely, which is established by IR analysis of aliquots. The solvent was removed under reduced pressure, and the residue mixed with 50 ml of chloroform was filtered through silica gel G 25 in chloroform into a 60 ml sintered glass funnel. The product was eluted with chloroform until 200 ml of chloroform was collected. The chloroform was removed under reduced pressure to give 0.632 g of 7-methoxy-7-aminocephalosporic acid benzhydryl ester, which was acylated directly without purification. The starting compound was prepared according to the procedures described in 35 Example 1, starting from 7-aminocephalosporanic acid benzhydryl ester.

B. 7-Methoxy-7- (2-thienylacetamido / cephalosporic acid benzhydryl ester)

0.632 g of тоmethoxy-7-aminocephalosporanic acid benzhydryl ester is mixed with 25 ml of methylene chloride and cooled to 0 ° C, 0.6 ml of 2-thienyl 45 acetyl chloride is added dropwise over 30 seconds, followed by 0.6 ml pyridine 60 seconds later. The reaction mixture was stirred at 0 ° C for 15 min and poured into crushed ice. The mixture was stirred and the organic layer was separated and washed once with 20 ml of water, once with 20 ml of sodium bicarbonate and once more with 20 ml of water. The methylene chloride was dried and evaporated to dryness to give 1.417 g of crude product. This material was applied to a column of 60 g of silica gel in benzene and the column was eluted with benzene, collecting fractions of 100 ml followed by 300 ml of methylene chloride / benzene 1: 1 in 3 fractions and 500 ml of methylene chloride in 5 fractions. The product was separated from the column by elution with 400 ml of chloroform in 4 fractions to give 0.592 g. This material was dissolved in 25 ml of methylene chloride and stirred at room temperature with 20 ml of a solution of 0.120 g of sodium bicarbonate in water for half an hour. The layers were separated and the organic layer was washed with water, dried and evaporated to dryness, yielding 0.420 g of 7-methoxy-7- (2-thienylacetamido) -phalosporanic acid benzohydryl, which said 1 spot on a thin layer chromatographic crying. IR: 3.05 μ / ΝΗ /; 5.62 μ / β-lactamine C-0 /; 5.75 μ / ester C = 0 /; 5.92µ (amide C = 0);

NMR: 2.6-3.2 Tau / C ₆ H ₅ and

protons /; 4.94Tay (S, 6H), 5.05 tau g (CH ₂ -OAc); 6.11 tau / S

6.52 Tau (S, OCH ₃ ), 6.64 _T ay (-CH ₂ -S);

7.99 tau / S, CH ₃ - C = 0 /

C. 7-Methoxy-7/2-thienylacetamido / cephalosporanic acid sodium salt

0.420 g of 7-methoxy-7- (2-thienylacetamido / ce30 phalosporanic acid benzohydryl ester was dissolved in 3.5 ml of anisole and treated with 10 ml of trifluoroacetic acid at room temperature for 10 minutes. The trifluoroacetic acid was removed and the anisole removed. under reduced pressure, keeping the temperature below 40 ° C, and the residue mixed with 25 ml of chloroform and treated with 20 ml of water containing 0.120g of sodium bicarbonate. The mixture was stirred for half an hour at room temperature and the organic phase was separated and washed with water. The combined aqueous phase was washed twice with methylene chloride and lyophilized to give 0.382 g of the 7-methoxy-7- (2-thienylacetamido) -cephalosporanic acid sodium salt as a brownish solid. IR: 5.65 μ / β-lactam /, 5.91 μ / amide carbonyl /; NMR: (DMSOD ₆ ): 2.65 tau (singlet) and 3.06 tau (doublet) (thienyl protons); 5.04 tau (singlet) and 3.06 tau (doublet) (thienyl protons), 5.04 tau (singlet, 6 N);

5.16 Tau, Sq / CH-O- £ CH ₃ /; 6, 19 tau

N ⁰ (singlet ^ B ^ CH 6.65 tau / singlet, OCH ₃ ); 6.77 tau (-S-CH ₂ ); 8.01 tau / CH _{3 -} C = O /.

Example 4

7-Methoxy-7- (2-thienylacetamido) -3-desacetoxy-3-pyridinecephalosporanoic acid thiocyanate

0.100 g of the 7-methoxy-7- (2-thienylacetamido) -cephalosporanic acid sodium salt is dissolved in 100 ml of water containing 50 ml of pyridine, 5 ml of 85% phosphoric acid and 0.475 g of potassium thiocyanate. The reaction mixture was stirred at 60 ° C for 5 h and cooled to room temperature. The reaction mixture was diluted with water to a total volume of 20 ml and extracted 5 times with 5 ml portions of chloroform. The chloroform was removed from the aqueous phase by evaporation in vacuo and the aqueous phase was then cooled to 0 ° C and acidified to pH 2. The mixture was left at 0 ° C for 2 h and the precipitated solid was removed by filtration and dried to give 0.015 g of thiocyanate of 7-methoxy-7- (2-thienylacetamido) -3-desacetoxy-3-pyridinecephalosporanic acid as a pale solid. IR:

4.83 μ / CNS— /, 5.62 μ / β-lactate /. R _f 0.61 / BAW 3: 1: 1, on paper /.

Example 5

A. 7-Methoxy-7- (2-thianaphthene-2-acetamido / cephalosporic acid benzhydryl ester

330 mg / 1.57 mm / benzothiophene-2acetyl chloride was added in 1 portion to a solution of 330 mg / 0.75 mm / 7-amino-7-methoxycephalosporanic acid benzhydryl ester in 10 ml of methylene chloride at 0 ° C. After one minute, 330 ml of dry pyridine was added and the homogeneous reaction mixture was stirred for 15 min at room temperature and poured into 10 ml of water. The organic layer was washed once with 3 ml of cold aqueous 5% sodium bicarbonate solution and once with 3 ml of cold water and dried over sodium sulfate. The solvent was removed in vacuo at room temperature and the crude product was chromatographed on

2.1 cm / outer diameter / column containing 30 g of silica gel with methylene chloride. All less polar impurities, including the azide from the previous step, were eluted with 400 ml of methylene chloride. The eluent was then replaced with chloroform to separate the product. All fractions containing the product were combined and washed with 5% sodium bicarbonate and water. The solution was dried over sodium sulphate and evaporated to dryness in vacuo to give 301 mg of 7-methoxy-7- (2-thiophthalene-2-acetamido / cephalosporanic acid 7-methoxy-7- (2-thianaphtheno) cephalosporanic acid ester. IR: 6.65 μ / β-lactam /, 5.78 μ / ester /, 5.95 μ / amide /. NMR: 8.02 Tau 5 (-OCHS,); 6.52 tau / -0CH7, 6.08 ^{00 11} z ^S tau (CH-C-NH-), 4.94 tau / HC (/,

5.0-5.2 tau <-CH ₂ OSSN ₃ ), 2.5 -3.1 tau 10 (aromatic protons). TLC: Silica gel G, 5% ethyl acetate / methylene chloride R = 0.37.

B. Sodium salt of 7-methoxy-7/2-cyanophen-2-acetamido / cephalos-15 tartaric acid

2.0 ml of anisole and 5.9 ml of trifluoroacetic acid were combined and added to 300 mg of 7-methoxy-7- (2-thiaphthene-2-acetate-20 mido / cephalosporic acid benzhydryl ester and the reaction mixture was stirred for a while for 10 min at room temperature. The excess anisole and trifluoroacetic acid was removed in vacuo and the residual anisole and trifluoroacetic acid removed by placing the flask directly on a high-vacuum pump for 15 min. The remaining dark oil was stirred in 15 ml of a mixture of benzene. / 1: 1 /. A small amount of insoluble rubber is dissolved by adding 5 ml of a 5% solution of sodium bicarbonate in water. The organic layer was washed with water and the combined aqueous layers were lyophilized to give 185 mg of the sodium salt of 7-methoxy-7- (2-thiophthalene).

2-acetamido / cephalosporanic acid in the form of a pale yellow powder. NMR: 6.18 tau (-CH ₂ CNH-), 8, 13 tau (-O = C-CH ₃ ), 6.52 tau (-OCH ₃ ).

Example 6

7-Amino-7-45 methoxycephalosporanic acid benzhydryl ester

500 mg of platinum oxide is added to a solution of 500 mg / 1.07 mmol / benzhydrol ester of 7-azido-7-methoxy cephalosporic acid in 50 ml of p-dioxane in a 250 ml round bottom flask. The reaction vessel was placed under hydrogen at room temperature and atmospheric pressure with vigorous stirring with a magnetic stirrer. After one hour, 500 mg of fresh platinum oxide was added and the reaction continued under the same conditions for another 3 hours. The dioxin was removed in vacuo at room temperature and the residue was stirred in 5 ml of chloroform. The catalyst was removed by passing the mixture through 15 g of silica gel G embedded in a sintered glass funnel. The product was eluted with 400 ml of chloroform using a vacuum. The chloroform was evaporated in vacuo to give 300 mg of 7-amino-7-methoxycephalosporic acid benzhydryl ester as a yellow oil.

Example 7

7-Methoxy-7- (p-guanidinophenylacetamido) cephalosporic acid benzhydrol ester hydrochloride

To a solution of 250 mg benzhydryl ester of 7-amino-7-methoxycephalosporanic acid dissolved in

1.5 ml of dry dimethylformamide are added in one portion while cooling and stirring 1.15 g of p-guanidinophenyl acetyl chloride and 0.15 ml of pyridine. The reaction mixture was stirred at 5 ° C for 5 min and at room temperature for another 10 min. The solution was diluted with 15 ml of methylene chloride and extracted 3 times with 15 ml of water. The methylene chloride solution was dried and evaporated to dryness in vacuo and the residue was chromatographed on 20 g of silica gel. It was eluted with chloroform then with a 1: 1 ethanol / chloroform mixture to give 100 mg of 7-methoxy-7- (p-guanidine-phenylacetamide / cephalosporic acid benzohydryl ester). TLC on silica gel using 1: 1 ethanol / chloroform showed one spot with R _f = 0.6. IR: 6.5 μ (elbow),

5.8 μ (ester).

B. 7-Methoxy-7- (p-guanidinephenylacetamide) -cephalosporic acid

A solution of 88 mg of the above ester and 0.7 ml of anisole dissolved in 1.8 ml of trifluoroacetic acid was kept at room temperature for 10 minutes. The solution was evaporated in a high vacuum pump for 5 minutes and the residue was treated with ether to a solid. After decanting the ether, the solid residue was stirred with 50 ml of water and filtered. The clear filtrate was lyophilized to give 40 mg of 7-methoxy-7- (p-guanidinephenylacetamide) -cephalosporic acid. Paper chromatography using butanol-acetic acid in water 3: 1: 1 showed a single spot with R _f 0.25, which gave a positive Sakaguchi test and activity against B. subtilys. IR: 5.7μ (ester, arm), 5.65 μ (lactam).

Example 8

7-Methoxy-7- (2-furylacetamido) -cephalosporanic acid sodium salt

7-Methoxy-7-azidocephalosporanic acid benzhydryl ester 1.00 g was hydrogenated in 100 ml of dioxane for one hour with 1 g of PtO ₂ , then for another 3 hours with another gram of PO ₂ . The solvent was removed in vacuo at a temperature below 30 ° C. The residue was mixed with chloroform and filtered through a layer of about 2.54 cm of silica gel (for thin layer chromatography), washed thoroughly with chloroform, a total volume of about 500 ml. Removal of the chloroform in vacuo gave 7-methoxy-7-aminocephalosporanic acid benzhydryl ester. 40 ml of methylene chloride is added and then 0.7 ml of furylacetyl chloride and then 1 ml of pyridine are added first at 0 ° C. After 25 min with stirring at 0 ° C, water was added and stirring continued for a few more minutes. The layers were separated and the organic portion was washed sequentially with 1% aqueous H ₃ PO ₄ , water and saturated aqueous sodium bicarbonate solution. After drying the methylene chloride solution with magnesium sulfate, filtration and evaporation of the solvent, the -7- (2-furylacetamide) -cephalosporanic acid benzhydryl ester is obtained. Purify by chromatography on 60g neutral silica gel (Bpinkmann 70-325 mesh ASTM) and elute with 4: 1 chloroform-ethyl acetate. The R _f value on the column is 0.69-0.45 and the TLC in the same system is 0.69-0.57, one spot. IR: (CHCI3 solution) has bands at 3.0 μ (Ν-Η), 5.61μ (βlactam), 5.76μ (esters) and 5.9μ (amide). The NMR spectrum with CDC1 ₃ has bands at 8.05 tau (3H, singlet, -COCH ₃ ), 6.65, 6.70 tau (2H, S-CH-), 6.55 tau (3H, singlet, -OCH). ₃ ), 6.32 tau (2H, singlet, furyl-CH ₂ -), 4.85, 5.05, 5.15, 5.35 tau (2H, AB quartet, I = 14 Hz; -CH ₂ OAc) ), 4.97 tau (1H, singlet, C ₆ ~ H), 3.72 tau (2H, multiplet, furyl β-Η), 3.09 tau (1H, singlet, -CH ₂ ), 2.7 tau (6H, multiplet, phenyl and furyl a-H).

0.57 g of the compound was treated at 0 ° C for 5 min with 0.8 ml of anisole and 4.0 ml of trifluoroacetic acid. Trifluoroacetic acid and anisole were removed at a temperature below 30 ° C in vacuo, 2 ml of anisole was added and evaporated as above. The residue was mixed with several ml of water containing 0.1 g of sodium bicarbonate and lyophilized into a powder, which was carefully washed with ether and dried, yielding 0.45 g of sodium salt of 7-methoxy-7- / 2- furylacetamide / -cephalosporanic acid. IR: (CHCl ₃ solution) has bands at 3.1μ (broad, Ν-Η), 5.67 μ (β-lactam), 5.75 μ (esters), 5.92 μ (amide) and 6, 18 μ (COONa). The NMR spectrum in D ₂ O has bands at 7.92 tau (3H, -COCH ₃ ), 6.60, 6.66 tau (2H,

S-CH-), 6.47 tau (3H, -OCH ₃ ), 6.20 tau (2H, furyl-CH ₃ ), 5.38 tau (HDO), 5.13 tau (2H, -CH ₂ OAc) ), 4.88 tau (1H, C ₆ -H), 3.63 tau (2H, furyl β-Η), 2.53 tau (1H, furyl α-H). The B spectrum in pH 7 buffer has L _{MaK (} 263 mm E% = 141. R _f of TLC (silica gel, acetone - AcOH 9: 1) is 0.68.

Example 9

A. 7-Methoxy-7-tetrazole benzhydryl ester and cetane and d-cephalosporic acid

To a solution of 1.17 g of 7-amino-7-methoxycephalosporic acid benzhydryl ester in 100 ml of methylene chloride cooled at 0-5 ° C in an ice bath was added 1.78 ml of pyridine with stirring and then a cooled solution of 1 , 17 g of tetrazolylacetyl chloride in 100 ml of methylene chloride. The mixture was allowed to react for 10 minutes at 0-5 ° C. Then shake 200 ml with pH 2 phosphate buffer. The methylene chloride layer was dried and evaporated. 1.2 g of the crude product was eluted through 48 g of silica gel using chloroform as eluent. The desired product was eluted with 2% methanol / chloroform to give 450 mg of 7-methoxy-7-tetrazolylacetamidcephalosporanic acid benzhydryl ester. NMR: 8.0 tau (acetyl singlet), 6.53 tau (SCH ₂ ),

4.84 tau (6H, singlet), 3.0 tau (singlet, benzhydryl CH), 2.6 tau (singlet, aromatic), 1.1 tau (tetrazole H). TLC on silica gel in 5% methanol / chloroform R _f = 0.28.

B. 7-Methoxy-7-tetrazolylacetamidocephalosporanoic acid sodium salt

A mixture of 680 mg of 7-methoxy-7-tetrazolylacetamidocephalosporanoic acid benzohydryl ester, 4.4 ml of anisole and 12.2 ml of trifluoroacetic acid was stirred at room temperature for 8 min. The excess acid was evaporated under reduced pressure and the residue was washed twice with water. tetrachloromethane and then three times with hexane. The solid was dissolved in ethyl acetate, adjusted to pH 5.8 with sodium bicarbonate solution, extracted with water, freeze dried to give 420 mg of 7-methoxy-7-tetrazolylacetamidocephalosporanoic acid sodium salt.

Example 10

7-methoxy-7- (1 / 1H) -tetrazolylacetamide / -3- (5-methyl-1,3,4-thiadiazolthiomethyl) cefam-4-carboxylate

A mixture of 420 mg of 7-methoxy-7-tetrazolylacetamidcephalosporic acid sodium salt, 168 mg of 2-methyl-5-mercapto-1,3,4-thiadiazole and

16.8 ml of phosphate buffer with pH 6.4 was heated in a steam bath for 1/2 hour. The reaction mixture was adjusted to pH

4.85 and extracted with 25 ml of ethyl acetate. The aqueous layer was adjusted to pH

1.8 with 2.5 N hydrochloric acid solution and extracted with ethyl acetate. Ethyl acetate was removed under reduced pressure and the residue was washed three times with ethanol to remove traces of ethyl acetate. It is then mixed with ethanol and water, brought to pH 6.5 with sodium bicarbonate solution, treated with charcoal and filtered. The ethanol was evaporated and the aqueous layer was dried in cold to give 210 mg of sodium methoxy-7- (1 (1H) -tetrazolylacetamide) -3- (50 methyl-1,3,4-thiadiazole34 thiomethyl) -cephem- 4-carboxylic acid. UV λ H ₇ O 273 E% 155. NMR: 7.28 tau (singlet, methyl in thiadiazole), 6.42 tau (methoxy group singlet), 0.74 tau (singlet, tetrazole H).

Example 11.

A. 7-Aminodesacetoxycephalosporanic acid benzhydryl ester in 1.5 water in a 5-liter three-necked flask equipped with an addition funnel is added 3.5 g of boron trifluoride in 80 ml of dioxane. The mixture was stirred for 1 h (pH 2.2), 1.5 L of acetone was added and the mixture was stirred for 10 min (pH 2.4). Diphenyldiazomethane 31.4 g, 0.162 mol in 85 ml of acetone was added dropwise, with good stirring for 4 h, during which time the suspension was diluted. The mixture was filtered and the solid was air-dried, washed well with chloroform to give

3.8 g of recovered starting product.

These 3.8 g of starting material are recycled using 750 ml of water, 1.2 g of BF ₃ in 30 ml of dioxane, 750 ml of acetone and 11 g of Ph ₂ CN ₂ in 30 ml of acetone as described above.

The filtrates from both cycles were evaporated under reduced pressure to remove the acetone. The aqueous residues were extracted with chloroform (3x250 ml, 3x150 ml). After drying the chloroform extracts with magnesium sulfate and evaporating, 45 g of crude product are obtained, which is chromatographed on 125 g of silica gel using chloroform and a mixture of chloroform-methanol (1-3%) as eluent. Fractions 3-8 were treated separately with ether. Crystals were obtained from fractions 4-7 - the first amount, mp 133-143 ° C, 5.58 g; the second amount, mp 95-125 ° C, 0.52g.

The filtrates and adjacent fractions of the above chromatography were combined and re-chromatographed as indicated above. Fractions 4-12 were treated with ether. Fraction 9 crystallized well, yielding a third amount, mp 133-142 ° C, 1.47 g.

The total yield of 7-aminodesacetoxycephalosporic acid benzhydryl ester is 7.57 g. The starting material is extracted from the aqueous phase by concentration in vacuo to remove water and bring the pH to 3.7. The starting product precipitates with stirring. Filtration, washing the precipitate with water and acetone and drying afforded 3.19 g of recovered starting material.

The 7-amino-3-desacetoxycephalosporanic acid benzhydryl ester is characterized by a melting point of 146-150 ° C, IR, NMR and elemental analysis. Calculated for C ₂ H ₂₀ N ₂ O ₃ S: C 66, 30; H, 5.30; N, 7.36. Found: C, 66.24; H, 5.55; N, 7.07.

The 7-aminodesacetoxycephalosporanic acid used as starting material was obtained by hydrogenolysis of 7-aminocephalosporanic acid using a barium sulphate catalyst according to known procedures.

B. 7-Azido7-bromo-3-desacetoxycephalosporanoic acid benzhydryl ester

A mixture of 7.57 g of 7-aminodesacetoxycephalosporanic acid 7.7-aminodesacetoxycephalosporic acid ester, 14.7 ml of 2 N sulfuric acid, 400 ml of water and 400 ml of methylene chloride was stirred for one hour in a stoppered ice-cold flask. The layers are separated while cold. The aqueous layer was washed with methylene chloride and the combined CH ₂ Cl ₂ layers were dried with magnesium sulfate and evaporated under reduced pressure at <30 ° C to about 125 ml of a solution of 7-diazocephalosporic acid benzhydryl ester.

During the above reaction, the mixture was stirred, triethylammonium azide and bromazide were prepared. A mixture of 11.7 g of sodium azide,

9.7 ml of concentrated sulfuric acid, diluted to 40 ml with water, and 190 ml of methylene chloride were stirred for 30 min in a closed ice bath system. The layers were separated in cold and the aqueous residue was washed with 10 ml of cold methylene chloride. The CH ₂ Cl ₂ layers were combined, dried with magnesium sulfate, kept in an ice bath and separated into two equal parts. To this was added 4.9 ml of triethylamine; 4.93 g of N-bromosuccinimide were added to the other. Both solutions are kept in an ice bath until used.

The 7-diazo compound in about 125 ml of methylene chloride in a 500 ml round-bottom flask equipped with a tube of calcium chloride and magnetically stirred is cooled in a bath of solid CO ₂ / acetone to -40 ° C (internal temperature). The bath temperature was maintained at -40 ° C to -50 ° C during the reaction. The triethylammonium azide solution was added at once. The bromazide solution was added over 5 min at -30 ° C to -25 ° C. The flask was removed from the bath and allowed to rise to 0 ° C for 20 min.

10 g Ν ₂ ΗΡΟ ₄ in 300 ml of water are mixed with the reaction mixture and the layers are separated. The methylene chloride layer was dried with magnesium sulfate and evaporated to give 10.3 g of crude product which was chromatographed on 125 g of silica gel using benzene as eluent. When the yellow color reaches the bottom of the column, the following fractions are collected: fraction 1-1.38 g; fraction

2-1,3 g; total yield - 5.1 g. The two fractions crystallized immediately.

An analytical sample of 7-azide-7-bromo-3-desacetoxycephalosporanic acid benzhydryl ester, melting point 122 ° C, is characterized by IR, NMR and elemental analysis. Calculated for C ₁₉ H ₁₇ N ₄ O ₃ S: C 51.97; H, 3.53; N, 11.54. Found: C, 53.23; H, 3.59; N, 11.63.

C. 7-Azide7-methoxy-3-desacetoxycephalosporanoic acid benzhydryl ester

A solution of 5.1 g (10.5 mmol) of 7-azide7-bromo-3-desacetoxycephalosporanic acid benzhydryl ester in 50 ml of methylene chloride and 200 ml of methanol is prepared. Pyridine 0.844 ml, 10.5 mmol, and 2.084 g silver fluoroborate 10.7 mmol in 10 ml methanol are added and the reaction mixture is stirred at 22 ° C for 16 1/2 hours. The mixture is filtered and the filtrate is evaporated under reduced pressure. The residue was chromatographed on 125 g of silica gel using benzene as eluent. Fraction 1 was collected when the yellow color reached the lower end of the column. Fractions 2-9 were combined and treated with methanol. Methoxyazide crystals have a melting point of 115-117 ° C, 3.156 g and are characterized by IR, NMR and elemental analysis. Calculated for C ₂₂ H ₂₀ N ₄ O ₄ S: C 60, 54; H, 4.62; N, 12.84. Found: C, 60, 42; H, 4.36; N, 12.93.

D. 7- (D-Azaidophenylacetamide) -7-methoxy-3-desacetoxycephalosporanoic acid benzhydryl ester

A mixture of 1.51 g of 7-azide-7-methoxy-3-desacetoxycephalosporanoic acid benzhydryl ester, 150 ml of dioxane and 1.5 g of platinum oxide was stirred under nitrogen for 1 hour. An additional 1.5 g of catalyst was added and the mixture was hydrogenated for an additional hour under reduced pressure (<35 ° C). The residue was mixed with chloroform and passed through silica gel diatomaceous earth (1: 1) in a 120 ml sintered funnel. About 500 ml of eluent was collected. The filtrate was evaporated and the process repeated in a 60 ml sintered funnel.

The filtrate, about 500 ml, was evaporated under reduced pressure and the residue was dried with nitrogen to give 2 g of 7-amino-7-methoxydesacetoxycephalosporic acid benzhydryl ester.

7-Amino-7-methoxydesacetoxycephalosporanic acid benzhydrol ester was mixed with 50 ml of methylene chloride and stirred with an ice bath magnet. To the solution was added

1.65 g of D-α-azidophenylacetyl chloride. After 3 min, 1.4 ml of pyridine was added. The solution was stirred in an ice bath for 25 min, poured into 50 ml of ice water and the layers separated. The methylene chloride layer was washed with 40 ml of dilute sodium bicarbonate solution and 50 ml of water and dried overnight in magnesium sulphate refrigerator. The mixture was filtered, evaporated and dried to give 2.0 g of product. The product was chromatographed on 100 g of silica gel using benzene and benzene / chloroform as eluent. Fractions 16-20 benzene / chloroform 1: 3 contained the product 7- (O-a-azidophenylacetamide) 7-methoxy-3-desacetoxycephalosporanate benzhydryl ester (843 mg), which was characterized by IR and NMR.

E. 7- (O-a-Azidophenylacetamide) -7methoxy-3 - desacetoxy cephalic acid

A solution of 708.4 mg of 7- (O-α-azidophenylacetamide) -7-methoxy-3-desacetoxycephalosporanic acid benzhydryl ester in 2 ml of anisole is cooled in an ice bath and 8 ml of trifluoroacetic acid are added. The reaction solution was kept at 0 ° C for 10 min, shaking from time to time. The solution was evaporated under reduced pressure, washed with anisole and evaporated twice. The residue was mixed with 30 ml of methylene chloride and extracted with 3x5 ml of saturated aqueous sodium bicarbonate solution. The combined aqueous phases were washed with 2x5 ml of methylene chloride and the pH adjusted to 1.8 with 2.5 N HCl and then extracted with 4x10 ml of ethyl acetate. The extracts were dried over magnesium sulfate and evaporated under reduced pressure at <45 ° C, then applied directly to a pump. The product obtained (487.4 mg) was characterized by IR and NMR.

Example 12

7- (D-α-aminophenylacetamide) -Ίmethoxy-3-desacetoxycephalosporic acid

To an ice - cold solution of

487.4 g of 7- (N-α-azidophenylacetamide) 7-methoxy-3-des-cetoxy cephalic asporic acid in 6 ml of acetic acid and 9 ml of water was added 3.1 g of powdered zinc. The mixture was stirred in an ice bath for 10 min and then filtered. The solid was washed with 40 ml of water. The combined aqueous filtrates were saturated with hydrogen sulfide. After filtration, the filtrate was lyophilized to give 0.5 g of crude product. The crude product was dissolved in water and lyophilized twice more, yielding 400 mg of a product characterized by IR, NMR, electrophoretic mobility, reaction with ninhydrin and UV 262.5 mg, ε = 5770 /.

Example 13

A. 7- (D-azido-2-phenylacetamide) -7-methoxycephalosporanic acid benzhydryl ester

To a solution of 1 g of 7-amino-7-methoxycephalo-sporic acid benzohydryl ester in 25 ml of methylene37 chloride at 0 ° C was added 1.1 g of D-azidophenylacenyl chloride in 15 ml of methylene chloride and then 1 ml of pyridine. After stirring at 0 ° C for 15 min, the mixture was extracted with 2x5 ml of cold water, 3x5 ml of 1% aqueous phosphoric acid, 3x5 ml of saturated aqueous sodium bicarbonate solution and 2x5 ml of water. The methylene chloride solution was dried over magnesium sulfate, filtered and concentrated under reduced pressure to give 1.1 g of 7- / N-α-azido-benzhydryl ester.

2-Phenylacetamide) -7-methoxycephalosporanic acid as an oil. This product was chromatographed on 60 g of neutral silica gel and the product was eluted with chloroform. After evaporation of the solvent gave 600 mg of product whose R _f value of the column is from 0.069 to 0.047, and a thin layer chromatogram (silica gel, CHC1 ₃₎ is 0.25, single spot. IR spectrum: (CH1 ₃ solution) shows bands at 3.0 μ (Ν - H), 4.74 μ (azide), 5.6 μ (β-lactam), 5.76 μ (ester) and 5, 88 μ (amide). NMR spectrum: (deuterochloroform) has bands at 8.05 tau (3H, singlet, -CHOC ₃ ), 6.65 tau (pin, 2H, S -CH ₂ ~), 6.55 tau (3H, singlet, - OCH ₃ ), 4.85, 5.05, 5.15,

5.35 tau (2H, AB quartet, 1-14 Hz, -CH ₂ OAc), 4.97 tau (1H, singlet, CO-H), 4.89 tau (1H, singlet, / N ₃ / CH / , 3.09 tau (1H, singlet, -CHO ₂ ), 2.65, 2.75 tau (15H, phenyl).

D. 7- (N-α-azido-2-phenylacetamido) -7-methoxycephalosporic acid

The product obtained according to Example A mentioned above (600 mg) was treated for 5 min at 0 ° C with 1 ml of anisole and 5 ml of trifluoroacetic acid. The resulting reaction mixture was evaporated at 30 ° C, at 0.1 mm pressure and then treated twice with anisole and evaporated again. The residue thus obtained was dissolved in 25 ml of methylene chloride and extracted with 4x3 ml of methylene chloride, brought to pH 1.8 1 with 5% phosphoric acid and extracted with 3x10 ml of ethyl acetate. The ethyl acetate solution was dried with magnesium sulfate and evaporated to give 370mg of 7-fluoro-azido-2-phenylacetamido / -7methoxycephalosporanic acid. IR (CHCl ₃ ): 3-4 μ (COOH),

4.74 μ (azide), 5.62 μ (β-lactam), 5.75 μ (ester), 5.85 μ (amide), about 8 μ (acid C = 0); NMR spectrum (CDCI ₃ ): 7.92 tau (3H, singlet, -CHOC3), 6.65 tau (2H, doublet, S-CH ₂ ~), 6.51 tau (3H, singlet, -OCH ₃ ), 4.96 tau (2H, doublet, -CH ₂ OAc), 4.92 tau (1H, singlet, C ₆ ~ H), 4.83 tau (1H, singlet, f (N ₃ ) C-H), 2 , 55 tau (5H, phenyl).

C. 7- (N-α-amino-2-phenylacetamido) -7-methoxycephalosporanic acid

To a solution of 620 mg of 7- (O-α-azido-2-phenylacetamido) -7-methoxycephalosporanic acid in 6.2 ml of acetic acid and 9 ml of water was added

3.1 g of zinc powder and the solution was stirred for 6 min at 0 ° C. The zinc is filtered and washed with 60 ml of cold water. The combined filtrates were saturated at 0 ° C with hydrogen sulphide and filtered through a diatomaceous earth. The filtrate was washed with 3x50 ml ethyl acetate and the aqueous solution was warmed under reduced pressure to remove the dissolved ethyl acetate and finally lyophilized to give 480 mg of 7- (D-amino-2-phenylacetamido) -7-methoxycephalosporic acid white powder form. This product contains 1 equivalent of acetic acid and 2 equivalent of water and 2% of ammonia as acetate or an antibiotic salt. Spinkoanalysis showed 1.58 micromoles / mg phenylglycine (84% of theory 38). Weight analysis showed 17.8% weight loss at 110 ° C (99% of theory). Titration: inflection at pH 5.7 and 8.7, pH 1/2 = 7.0, EW 476 (theoretically for acetate dihydrate = 515). Electrophoresis at pH 7 shows a single spot as a mono anion. Calculated for C ₁₉ H ₂₁ N ₃ O _? . 2H ₂ O. AcOH + 2% NH ₃ : C 46.2; H, 5.7; N, 8.8; S 5.6. Found: C, 47.41; H, 4.99; N, 9.48; S, 6.36. Alkaline distillation and distillation titration showed 2% NH ₃ (pH 7 / buffer 7) / xmax. = 263; E% 116 (e = 6170). The NMR spectrum (100MHz, D ₂ O) has bands at 7.65 tau (AcOH, about 1 equivalent), 7.61 tau (singlet, -CHOC ₃ ),

6.17 tau (singlet, -OCH ₃ ), 6.02, 6.19, 6.45,6,62 tau (AB kW, J = Hz, S-CH ₂ -); 2,13 tau (singlet, phenyl); HOD at 5 tau screens the other protons. IR (Nuyol): 2.8-4.5 μ (ΝΗ ₃ +),

5.65 μ (β-lactam), 5.85 μ (ester), 6.2-

6.3 μ (COO-).

Example 14

A. 7- (2-Carboxy-2-phenylacetamido) -7-methoxycephalosporanoic acid benzhydryl ester

To a solution of 0.5 g of 7-amino-7-methoxycephalosporanate benzhydryl ester in 15 ml of methylene chloride was added monobenzhydryl phenylmalonyl chloride, obtained as described below, and then 0.5 ml of pyridine. The resulting reaction mixture was stirred for 30 min and then 12 ml of water was added. The aqueous mixture was stirred for 5 min and the layers were separated. The methylene chloride portion was washed with 2.5 N HCl, water, twice with aqueous sodium bicarbonate solution and saturated with sodium chloride. The solvent layer was then dried over magnesium sulfate, filtered and evaporated at 25 ° C under reduced pressure to give 0.695 g of 7- (2-carboxy-2-phenyl cetamido / -7-methoxycephalosporic acid benzhydryl ester). The ester was chromatographed on 50 g of neutral silica gel, eluting with chloroform to give 400 mg of product as a tan glass, IR: 3.03 μ (NH), 5.62 μ (β-lactam), 5 , 78μ (ester), 5.88 μ (amide). NMR / CCl ₄ ):

8.18 tau (3H, singlet, acetyl), 7.0 tau (2H, S-CH ₂ -), 6.80, 6.73 tau (3H doublet, diastereomeric OCH ₃ groups), 5.2 tau ( 4H, multiplet, -CH ₂ O ~, C ₆ proton, malonyl CH), 3.17 tau (2H, CHO ₂ ), 2.8 tau (20 H, aromatic), 2.0 tau (1H, amide NH) .

The phenylmalonyl chloride monobenzhydryl ester used above was prepared as follows. To a solution of 19.25 g of phenylmalonic acid in 165 ml of ethyl acetate was added a solution of 25 g of diphenyldiazomethane in 100 ml of ethyl acetate for 15 minutes at 15-20 ° C. The solution was stirred for another 10 minutes and 500 ml of water was added and 15 ° C was added 50% sodium hydroxide in sufficient quantity to alkalize the mixture. The solvent layer was separated from the mixture, extracted twice with sodium bicarbonate solution in water. The combined aqueous solutions were washed twice with ethyl acetate, cooled, acidified with hydrochloric acid and extracted three times with ethyl acetate. The ethyl acetate extracts were washed twice with water, once with saturated sodium chloride solution and then dried over magnesium sulfate. The solvent was then evaporated at a temperature below 25 ° C under reduced pressure to an oil which was crystallized from 200 ml of 1: 3 ether in petroleum to give 20.9 g of phenylmalonic acid monobenzhydryl ester at mp 119.5 ° C - 122 ° C. To a suspension of 0.7 g of this monoester in 2.5 ml of water was added 2.10 ml of 0.962 N sodium hydroxide. The solution was stirred for 3 min, filtered and lyophilized to give the monoester sodium salt. To this sodium salt was added 5 ml of benzene and the suspension was treated at 0 ° C with

1.5 ml of oxalyl chloride. After 10 min at 0 ° C and 5 min at 25 ° C, the mixture was evaporated at a temperature lower than 25 ° C under reduced pressure and the residue was concentrated twice with tetrachloromethane. In a dry atmosphere, the product in 5 ml of tetrachloromethane was filtered and concentrated in vacuo to give monobenzhydryl phenylmalonyl chloride.

B. 7- (2-Carboxy-2-phenylacetamido) -7-methoxycephalosporanic acid disodium salt

7- (2-Carboxy-2-phenylacetamido) -7-methoxycephalosporanic acid benzhydryl ester (400 mg) was treated at 0 ° C for 2 min with 1.2 ml of anisode and 6 ml of trifluoroacetic acid. The resulting reaction product was then frozen, evaporated at low temperature and high vacuum, diluted with anisole and evaporated again at 25 ° C. The free acid-containing residue was stirred in 20 ml of one-molar sodium bicarbonate solution, washed 4 times with small amounts of methylene chloride, acidified with HCl, saturated with NaCl and extracted with 4x10 ml of ethyl acetate. The solvent extracts were washed twice with saturated sodium chloride solution, dried over magnesium sulfate, filtered and evaporated under reduced pressure to give 218 mg of 7/2-carboxy-2-phenylacetamido) -7-methoxycephalosporic acid in the form of yellow syrup. The latter was dissolved in 5 ml of water containing mg of sodium bicarbonate and lyophilized. The lyophilized residue was dissolved in 5 ml of water, filtered and lyophilized again to give 182 mg of 7 (2-carboxy-2-phenylacetamido) -7-methoxycephalosporic acid disodium salt. The product thus obtained is mixed with 550 mg of the same product obtained from a second charge and dissolved in 10 ml of water. The resulting solution was filtered and lyophilized to give 647 mg of product. UV / pH 7 / λ 265 nm ε = 6300. NMR / D, O /: 7.90 tau (3H, singlet, acetyl), 6,70,6,54 tau (2H, 2AB quartets, diastereomeric S-CH ₂ -), 6.50, 6.38 tau (3H, 2 singlets, diastereomeric OCH ₃ ), 5.22 tau (2H, broad singlet, -CH2O-), 4.85 tau (1H, singlet, C ₆ proton) , 2.55 tau (5H, singlet, aromatic). The wide HOD peaks cover the adsorption of the malonyl proton. The wide HOD peaks cover the adsorption of the malonyl proton. At pH 7 electrophoresis, the product moves like a dianion, with only a spot. The pH of 10% aqueous solution was 8.8. Calculated for C ₂₀ H _lg N ₂ SO ₄ Na ₂ + Na ₂ CO ₃ + 0,4NaHCO ₃ + H ₂ O; C, 38.59; H, 3.09; N, 4.21; S, 4.81; ash (as Na) 15.19. Found: C, 38.99; H, 3.10; N, 4.17; S, 4.91; ash (as Na) 15.3.

Example 15

A. 7-Azido7-benzyloxycephalosporanoic acid benzhydryl ester

A mixture of 2.4 g of 7-azido-7-bromocephalosporanic acid benzhydryl ester prepared as described in Example 2C and 1.5 g of silver tetrafluoroborate in 10 ml of benzyl alcohol was stirred at room temperature for 3 h. was diluted with 300 ml of ether and the silver salt was removed by filtration. The filtrate was washed with aqueous sodium bicarbonate solution, dried over sodium sulfate and evaporated under reduced pressure. The excess benzyl alcohol is removed on a high vacuum pump with magnetic stirring for 18 h and the residue is chromatographed on silica gel. Elution with hexane, followed by increasing concentrations of methylene chloride in hexane and evaporation of the fractions containing the product, gave the 7-azido-7-benzioxycephalosporanic acid benzhydryl ester.

B. 7-Benzyloxy-7-phenylacetomidocephalosporanoic acid benzhydryl ester

A solution of 1.2 g of 7-azido-7-benzyloxycephalosporic acid benzhydryl ester in 25 ml of dry ethyl acetate is hydrogenated in the presence of

1.2 g catalyst 10% palladium on charcoal for 18 h at room temperature. The catalyst was removed by filtration and phenylacetic anhydride (1.0g) was added to the filtered solution and the mixture was maintained for 45 minutes at room temperature. The resulting reaction mixture was diluted with 200 ml of ether, 200 ml of phosphate buffer pH 7 was added and the mixture was stirred vigorously for half an hour. The ether layer was separated, washed with water, dried and evaporated under reduced pressure. The residue was chromatographed on 50 g of silica gel, eluting with 5% ethyl acetate in methylene chloride to give 7-benzyloxy-7-phenylacetamidocephalosporanic acid benzohydryl ester upon evaporation.

C. 7-Benzyloxy7-phenylacetamidocephalosporanoic acid sodium salt

A solution of 0.5 g of 7-benzyloxy-7-phenylacetate benzhydryl ester of docetalosporanic acid in 3.5 ml of anisole is treated with 10 ml of trifluoroacetic acid at room temperature for 10 minutes. The trifluoroacetic acid and the anisole are removed under reduced pressure. lower than 40 ° C and the residue is mixed with 25 ml of chloroform and treated with 20 ml of water containing 0.15 g of sodium bicarbonate. The mixture was stirred for half an hour at room temperature and the organic phase was washed twice with methylene chloride and the aqueous layer and the combined washings were combined and lyophilized to give the 7-benzyloxy-7-phenylacetamidocephalosporic acid sodium salt.

Example 16

A. 7-Azido7-methylmercaptocephalosporanic acid benzhydryl ester

To a solution of 2 g of 7-diazocephalosporanic acid benzhydryl ester prepared as described in Example 2B, in a solution of 100 ml of methylene chloride at -75 ° C was added a solution of 1 ml of methylsulphenyl chloride in 10 ml of methylene chloride as vigorous solution. . Nitrogen is released immediately. After the addition of the reagent for a period of about 10 minutes, the mixture was gradually brought to - 5 ° C for about half an hour. Saturated sodium bicarbonate solution was added, the organic layer was separated and washed with water. After drying over sodium sulfate, the solvent is removed at room temperature under reduced pressure to give 7-chloro-7-methylmercaptocephalosporanic acid benzhydryl ester. This product was stirred with 20 ml of a solution of trimethylammonium azide in methylene chloride at 0 ° C -5 ° C under nitrogen for 1 1/2 hours. The solution was then washed with cold saturated sodium bicarbonate solution, water and dried over sodium. sulfate. Evaporation of the solvent gave crude 7-azido-7-methylmercaptocephalosporanic acid benzhydryl ester.

B. 7-Methylmercapto-7-phenylacetamidocephalosporanoic acid benzhydryl ester

A solution of 1.2 g of 7-azido-7-methylmercaptocephalosporanic acid benzhydryl ester in 25 ml of dry ethyl acetate containing 0.8 ml of diisopropylethylamine was hydrogenated in the presence of 1.2 g of 10% palladium catalyst on charcoal for 18 hours. h at room temperature. Phenylacetic anhydride (0.1 g) was added and the mixture was kept at room temperature for 45 min. The mixture was diluted with 200 ml of ether, 200 ml of phosphate buffer at pH 7 was added and the mixture was stirred vigorously for half an hour. The ether layer was separated, washed with water, dried and evaporated to give 7-methylmercapto-7-phenylacetamidocephalosporanoic acid benzhydryl ester.

C. Sodium salt of 7-methylmercapto-7-phenylacetamidocephalosporanic acid

A solution of 0.4 g of 7-methylmercapto-7-phenylacetamidocephalosporanoic acid benzhydryl ester in

3.5 ml of anisole is treated with 10 ml of trifluoroacetic acid at room temperature for 10 minutes. The trifluoroacetic acid and anisole are removed under reduced pressure while maintaining the temperature below 40 ° C and the residue is stirred in 25 ml of chloroform. and treated with 20 ml of water containing 0.12 g of sodium bicarbonate. The mixture was stirred for half an hour at room temperature and the organic phase was separated and washed with water. The combined aqueous phase was washed twice with methylene chloride and then lyophilized to give the 7-methylmercapto-7-phenylacetamidocephalosporanoic acid sodium salt.

Example 17

A. 7-Azido-7-ethoxycephalosporanic acid benzylhydryl ester

To a solution of 217 mg of 7-azido-7-bromocephalosporanic acid benzhydryl ester in 15 ml of absolute ethanol was added 31.6 μΐ of pyridine and 78 mg of silver fluoroborate and the mixture was stirred at room temperature for 2 h, protected from light and moisture. The mixture was evaporated to dryness in vacuo and the residue was chromatographed on 20 g of silica gel. Elution with chloroform afforded 121 mg of 7-azido-7-ethoxycephalosporanic acid benzhydryl ester, mp 144.5-145 ° C.

B. 7-Ethoxy-7-thienylacetamidocephalosporanoic acid benzhydryl ester

A solution of 320 mg of 7-azido-7-ethoxycephalosporic acid benzhydryl ester in 30 ml of dioxane was stirred with 320 mg of platinum oxide at room temperature under hydrogen atmosphere for 1 hour. An additional amount of 320 mg of catalyst was added and hydrogenation continued 5 h. The mixture was evaporated in vacuo to dryness and the residue was stirred in chloroform and filtered through 4.54 cm silica gel and evaporated to give 7-amino-7-ethoxycephalosporanic acid benzhydryl ester as a yellow oil. It is dissolved in methylene chloride and 0.2 ml of thienylacetyl chloride and 0.2 ml of dry pyridine are added to the cooled solution with stirring. The mixture was stirred for 15 min at 0 ° C and then poured onto ice and the methylene chloride layer was separated, washed with aqueous sodium bicarbonate solution, dried and evaporated. The residue was chromatographed on 20 g of silica gel. Elution with chloroform afforded 120 mg of 7-ethoxy-7-thienylacetamidocephalosporanoic acid benzhydryl ester. TLC: 2% MeOH in methylene chloride R _f 0.73. IR:

5.65 μ (elbow); 5.85, 5.95, 6.0 (amide). NMR: 8.82 tau (CH ₂ CH ₃ ), 8.0 tau (singlet, OCHOS ₃ ), 6.64, 6.67 tau (doublet, SCH ₂ ), 6.14 tau (singlet, thienyl CH ₂ ) , 4.96 tau (singlet, 6H).

C. 7-Ethoxy-7-thienylacetamidocephalosporanoic acid sodium salt

A solution of 0.9 g of 7-ethoxy-7-thienylacetamidocephalosporanic acid 7-ethoxy-7-thienylacetamphalosporanoic acid in 6.35 ml of anisole and 17.1 ml of trifluoroacetic acid was kept at room temperature for 5 min and then evaporated under high vacuum. The residue was stirred with water and methylene chloride and the pH was adjusted to 5.8 by the addition of sodium bicarbonate solution. The aqueous layer was evaporated and acidified to pH 2 and the rubber residue was filtered off. The latter was dissolved in butyl acetate, filtered and the filtrate was stirred with water with the addition of sodium bicarbonate solution to pH 6.5. The aqueous extract was evaporated and lyophilized to give 0.2 g of 7-ethoxy-7-thienylacetamidocephalosporanic acid sodium salt. Circular paper chromatography on butanol-ethanol-water (4: 1: 5) gave a spot with R _f 0.55. Analysis calculated: C, 46.74; H, 4.14; N, 6.00. Found: C, 47.73; H, 4.09; N, 5.73.

Example 18

A. 7- (dl-Fluorophenylacetamido) -7-methoxycephalosporanoic acid benzhydryl ester

To a solution of 0.9 g of 7-amino-7-methoxycephos losporanic acid benzhydryl ester in 45 ml of methylene chloride at 0 ° C was added with stirring 0.96 ml of d, 1-fluorophenylacetyl chloride. After 1 min, 0.96 ml of pyridine was added and the mixture was stirred for 15 min. Then the reaction mixture was poured into ice, shaken and the methylene chloride layer was separated. The aqueous layer was extracted twice with methylene chloride. The combined aqueous layers were washed with aqueous sodium bicarbonate solution, then with water and dried over sodium. The solvent was evaporated, leaving 1.2 g of 7- (d, 1-fluorophenylacetamido) -7-methoxycephalosporanic acid benzhydryl ester. The product in methylene chloride solution was chromatographed on 90 g of silica gel. The silica gel column was washed sequentially with 1000 ml of methylene chloride, 500 ml of methylene chloride containing 5% chloroform, and then with 300 ml portions of ethylene chloride containing 10%, 25% and 50% chloroform, respectively. The combined fractions of 50% chloroform eluates were evaporated under reduced pressure to give 370 mg of product. Analysis: Calculated: C, 63.56; H, 4.83; N, 4.63. Found: C, 62.71; H, 5.02; N, 4.38. TLC in 5% ethyl acetate in methylene chloride showed two spots with R _f 0.406 and 0.54.

B. 7- (01-α-Fluorophenylacetamide) -7-methoxycephalosporanic acid sodium salt

To a solution of 360 mg benzhydryl ester of 7- (d 1 -α-fluorophenylacetamide) -7-methoxycephalosporanic acid in 2.44 ml of anisole was added

6.9 ml of trifluoroacetic acid and the reaction mixture was left for 15 min. The mixture was evaporated under reduced pressure and a mixture of water and methylene chloride was added to the residue. The pH of the mixture was adjusted to 6.8 and the aqueous layer after drying in cold gave sodium 7- (d

1-a-fluorophenylacetamide) -7-methoxycephalosporanic acid, R _f in butanol-ethanol-water 4: 1: 5 0.41 and 0.60.

Example 19

A. 3-Methyl-7β-trichloroethoxycarboxamidodecephalosporic acid

To a solution of 21.4 g of 3-methyl-7-aminodecephalosporanic acid in 250 ml of water at pH 8.7, corrected with 15% sodium hydroxide solution, was added 21.2 g of trichloroethoxycarbonyl chloride for half an hour. maintained at 20 ° C and pH maintained at 8.7 using a pH stat. The reaction mixture was stirred for another half hour at this temperature and then the pH was adjusted to 2 and extracted with ethyl acetate using three extractions. The ethyl acetate layer was washed twice with water, dried over anhydrous sodium sulfate and evaporated under reduced pressure to give 3-methyl-7-trichloroethoxycarboxamidodecephalosporic acid.

The 3-methyl-7-aminodecephalosporanic acid used as a starting material in this example was prepared by catalytic reduction of cephalosporin C, followed by hydrolytic removal of the 5-aminoadipoyl moiety as described in US Patent 3,129,224.

B. 3-Methyl-7 β-trichloroethoxy carboxamidodecephalosporanoic acid p-methoxybenzyl ester

3-Methyl-7β-tetrachloxybasamidodecephalosporanova 22.38 g and triethylamine 14 ml were stirred in 200 ml acetone for 45 min and p-methoxybenzyl bromide 10.1 g was added to this mixture, stirring continued for 20 min at room temperature. The whole reaction mixture was then cooled in an ice bath and the precipitated triethylamine hydrobromide was removed by filtration. The filtrate was evaporated under reduced pressure and the residue was extracted by shaking with a mixture of ether and aqueous sodium hydrogen phosphate. The ether layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the 3-methyl-7β-pyrrolexamidodecephalosporanoic acid p-methoxybenzyl ester.

C. 3-Methyl-7-aminodecephalosporanoic acid p-methoxybenzyl ester

The protective trichloroethoxycarbonyl group was removed by the addition of ethyl acetate solution (50ml) containing p-methoxybenzyl ester.

3-Methyl-7β-tetrachloride to a vigorously stirred suspension of 5 g of zinc powder in 50 ml of 90% acetic acid in an ice water bath for 5-10 minutes. The temperature was maintained at 25-30 ° C. After 2 h, the solid was removed by filtration and washed with 100 ml of ethyl acetate. The combined filtrates were diluted with 200 ml of water, the layers were separated and the organic layer was washed with cold aqueous sodium bicarbonate solution, dried and concentrated under reduced pressure. The residue was chromatographed on silica gel 40 g using CHCl ₃ : MeOH mixture as eluent. 3-Methyl-7-aminodecephalosporanoic acid p-methoxybenzyl ester is obtained.

D. 3-Methyl-7-diazodecephalosporanoic acid p-methoxybenzyl ester

To a mixture of 1.6 g of sodium nitrite, 30 ml of water and 40 ml of methylene chloride with stirring at 0 ° C was added 0.8 g of 3-methyl7-aminodecephalosporanic acid p-methoxybenzyl ester, then a solution of 0.75 g p -toluenesulfonic acid in 5ml of water for pro44 for 5 min. The mixture was stirred at 0 ° C for 20 min and the organic phase was separated, washed with ice water, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. at room temperature to give 3-methyl-7-diazocephalosporanoic acid p-methoxybenzyl ester.

E. 3-Methyl-7-methoxy-7- (2-thienylacetamide / decephalosporanoic acid sodium salt

In the procedure described in Examples 2C, 2D, 3A, 3B and 3C, 3-methyl-7-diazocephalosporanoic acid p-methoxybenzyl ester is converted to the 3-methyl-7-methoxy (2-thienylacetamide) -decephalosporic acid sodium salt. .

Example 20

A. 3-Methyl-7P-trichloroethoxycarboxamidodecephalosporanoic acid o-nitrobenzyl ester

Replacing p-methoxybenzyl bromide in Example 19B with an equivalent amount of o-nitrobenzyl bromide and the resulting reaction product separated by the processes described, the corresponding o-nitrobenzyl ester was obtained.

B. 3-Methyl-7-aminodecephalosporanoic acid o-nitrobenzyl ester

The trichloroethoxycarbonyl protecting group was removed and the product removed using the procedures described in Example 19C. 3-Methyl-7-aminodecephalosporanoic acid is an o-nitrobenzyl ester.

C. 3-Methyl-7-diazodecephalosporanoic acid o-nitrobenzyl ester

The diazotization of the product from step B above and the separation of the dia-product were carried out according to the procedures described in Example 19E.

3-Methyl-7-azido-7-bromodecephalosporanoic acid o-nitrobenzyl ester

When the o-nitrobenzyl ester of

3-Methyl-7-diazodecephalosporanic acid was reacted with triethylammonium azide and bromazide, and the resulting reaction product was separated according to the procedures described in example 2C to give 3-methyl-7-azido-7-bromodecephalosporanic acid o-nitrobenzyl ester.

3-Methyl-7-azido-7 - methoxydicephalosporanic acid o-nitrobenzyl ester

The product of step E was reacted with methanol in the presence of silver trifluoroborate, and the resulting reaction product was separated as described in Example 1C to give the title compound.

F. 3-Methyl-7- (2-furylacetamide) -7-methoxydecephalosporanoic acid o-nitrobenzyl ester Hydrogenated and 3-amino compound obtained reacting with the resulting 7-amine compound the procedure described in Example 8 to give 3-methyl-7- (2-furylacetamide) -7-methoxydicephalosporanic acid o-nitrobenzyl ester.

G. 3-Methyl-7- (2-furylacetamide) -7-methoxydicephalosporanic acid sodium salt

A 0.5% solution of o-nitrobenzyl ester of 3-methyl-7- (2-furylacetamide) -7-methoxydicephalosporanic acid in dioxane was placed in a pyrex reaction vessel, completely vented and purged with helium. The solution was then irradiated with 3000 nM of light until the ester group as seen by TLC was removed. After evaporation of the solvent under reduced pressure, the residue was mixed with 15 ml of methylene chloride and stirred.

1/2 hour with a solution of one equivalent of sodium bicarbonate in 10 ml of water. The aqueous layer was lyophilized to give a mixture containing the 3-methyl-7- (2-furylacetamide) -7methoxydicephalosporanic acid sodium salt.

Example 21

A. 3-Benzoylthiomethyl-7 ^ -tert-butoxycarboxamidodecephalosporic acid

A solution of 35 g of 3-benzoylthiomethyl-7-aminodecephalosporanic acid in 250 ml of water was brought to pH 8.7 with a 15% solution of sodium hydroxide and maintained at the pH during the reaction by pHstat. To this solution at 20 ° C was added tert-butoxychloroformate for half an hour. After another half hour of stirring, the reaction mixture was brought to pH 2 and extracted with ethyl acetate using three extractions. The ethyl acetate solution was washed twice with water and dried over anhydrous sodium sulfate. After filtration, the solvent was removed under reduced pressure to give 3-benzoylthiomethyl-7-tert-butoxycarboxamidodecephalosporanic acid. This starting compound is a known derivative of the 7-aminocephalosporanic acid described in BE 650444.

B. 3-Benzoylthiomethyl 1-7 β-tert-butoxycarboxylic acid phenacyl ester and idodecephalosporic acid

3-benzoylthiomethyl-7β-tetra-butoxycarboxamidodecephalosporanoic acid 35.0 g and triethylamine 23 ml were stirred in 200 ml acetone for 45 min and 15.9 g of α-bromoacetophenone was added to the mixture and stirred for 20 minutes. The whole reaction mixture was then cooled in an ice bath and the triethylamine hydrobromide was removed by filtration. The filtrate was evaporated under reduced pressure and the residue was extracted by shaking with a mixture of ether and aqueous sodium hydrogen phosphate solution. The ether layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 3-benzoylthiomethyl-7β-butoxycarboxamidodecephalosporanic acid phenacyl ester.

C. 3-Benzoylthiomethyl-7-aminodecephalosporanoic acid phenacyl ester

Phenacyl ester of 3-benzoylthiomethyl-7β-tetra-butoxyacetic acid (10 g) was dissolved in 35 ml of anisole and 100 ml of trifluoroacetic acid at room temperature for 10 minutes. The solvents were removed under reduced temperature. S. The residue was mixed with water and neutralized with solid sodium bicarbonate. Chloroform was added and the chloroform layer was separated, washed with water and dried. The organic extract was concentrated under reduced pressure to give 3-benzoylthiomethyl] -7-aminodecephalosporanic acid phenacyl ester.

E. 3-Benzoylthiomethyl-7-diazodecephalosporanoic acid phenacyl ester

A mixture of 10 g of sodium nitrite, 4.5 g of 3-benzoylthiomethyl-7-aminodecephalosporanoic acid phenacyl ester, 300 ml of methylene chloride and 300 ml of water and ice was shaken in a separatory funnel. P-toluenesulfonic acid monohydrate (0.27 g) was added in three portions over 20 minutes. The mixture was shaken vigorously and the methylene chloride layer was separated, dried over sodium sulfate and evaporated under reduced pressure to 40 ml. The 3-benzoylthiomethyl-7-diazodecephalosporic acid phenacyl ester solution was used in the next step without isolating the intermediate.

E. 3-Benzoylthiomethyl-7 (2-furylacetamide) -7-methoxydicephalosporanic acid sodium salt

In the procedure described in Examples 1B and 1C, and using the 3-benzoylthiomethyl-7-diazodecephalosporanoic acid phenacyl ester instead of the 7-diazodecephalosporanoic acid benzyl ester, the diazo compound was converted to the 7-benzoylthiodoethiomethyl-methoxymethoxymethoxymethoxymethoxymethyl ester of phenyl ester acid. The latter product is then converted to the sodium salt of 3-benzoylthiomethyl-7- (2-furylacetamide) -7methoxydicephalosporanic acid according to the procedures described in Example 8.

Example 22

A. 3carbamoyloxymethyl-7-aminodecephalosporanoic acid trimethyl ester

A mixture of 0.5 g of 3-carbamoyloxymethyl-7-aminodecephalosporanic acid, 2 ml of hexamethyldisilazane and 8 ml of chloroform was stirred overnight at reflux temperature. The solvent and excess hexamethyldisilazane were removed under reduced pressure to give a residue containing 3-carbamoyloxymethyl-7-aminodecephalosporanic acid trimethylsilyl ester.

B. 3carbamoyloxymethyl-7-diazodecephalosporanoic acid trimethylsilyl ester

Mixture of trimethylsilyl ester of

3-Carbamoyloxymethyl-7-aminodecephalosporanic acid, 0.05 ml of trifluoroacetic acid and 5 ml of chloroform were stirred at 0 ° C and 0.2 ml of isoamyl nitrite was added. After stirring for one hour at 0 ° C, the resulting reaction mixture containing 3-carbamoyloxymethyl-7-diazodecephalosporanoic acid trimethylsilyl ester was used directly in the next step.

C. 3-Carbamoyloxymethyl-7-azido7-bromodecephalosporanoic acid

5 ml of nitromethane were added to the solution obtained in step B at 0 ° C. Then 1 ml of methylene chloride solution of trimethylammonium azide and 1.5 ml of methylene chloride solution of bromazide are added in rapid succession, both solutions being in significant excess of the required amount. The bromazide was discolored and nitrogen was removed. After about 5-10 minutes, 20 ml of 0.1 N sodium thiosulfate was added and the layers were separated. The aqueous phase was adjusted to pH 2 by the addition of a few drops of hydrochloric acid and extracted again with methylene chloride. The combined extracts were washed with water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 3-carbamoyloxymethyl-7azido-7-bromodecephalosporic acid.

The 3-carbamoyloxymethyl-7-acinodecephalosporanic acid used as a starting material in this example can be prepared as follows. 7-Aminocephalosporanic acid was reacted with tert-butoxycarbonylazide to give the 7β (tert-butoxycarbonyl) derivative by known methods. The derivative was intimately mixed with citrusacetyl stearase in aqueous phosphate buffer at pH 6, 5-7 for 15 h, and 3hydroxymethyl-7- (tert-butoxycarbonyl) aminodecephalosporanic acid was separated from the resulting reaction mixture. To 0.2 g of 3-hydroxymethyl-7P ~ (tert-butoxycarbonyl) aminodecephalosporanic acid suspended in 5 ml of acetonitrile, cooled to 0 ° C and maintained under nitrogen atmosphere, was added 0.15 ml of chlorosulfonyl isocyanate. The reaction mixture was stirred for 70 min and then evaporated under reduced pressure to dryness. The resulting residue was mixed with 10 ml of ethyl acetate and 10 ml of 0.1 N phosphate buffer. The pH of the aqueous layer was adjusted to about 1.6 and the mixture was stirred for 2 1/2 hours at room temperature. The pH was then adjusted to about 8 with aqueous tricalcium phosphate and the aqueous phase separated. The organic phase was re-extracted with 10 ml of phosphate buffer at pH 8. The combined aqueous phases were adjusted to pH 2.1 with hydrochloric acid and extracted twice with ethyl acetate. The ethyl acetate extracts were dried over sodium sulfate and evaporated under reduced pressure to give 0.055 g of residue. This residue was washed with ether to give 3-carbamoyloxymethyl-7β- (tert-butoxycarbonyl) aminodecephalosporic acid, which was recovered as a yellow solid.

3-Carbamoyloxymethyl-7P- (tert-butoxycarbonyl) aminodecephalosporanic acid (0.5 g) in 3.5 ml of anisole was stirred with 2 ml of trifluoroacetic acid at 0 ° C for 5 min. The resulting reaction mixture was evaporated under reduced pressure, leaving yields 3-carbamoyloxymethyl-7-aminodecephalosporanic acid, which is purified by crystallization from aqueous isopropanol.

Example 23

3-Picolinoylthiomethyl-7-diazodecephalosporanoic acid benzyl ester

Following the procedure described in Example 1A, the benzyl ester of

3-Picolinoylthiomethyl-7-aminodecephalosporanic acid, obtained by esterification of the free acid with benzyl chloride, is converted to the corresponding diazocompound 3-picolinoylthiomethyl-7-diazodecephalosporanic acid benzoyl ester. 3-picolinoylthiomethyl-7-aminodecephalosporanic acid is prepared by reacting 3hydroxymethyl-7-aminodecephalosporanic acid with thiopicolic acid according to the procedures described in GB N 1 101 423.

Example 24

3-Pyridinylmethyl-7-diazodecephalosporanoic acid benzhydryl ester

In the procedure described in Example 2A and 2B, 3-pyridinmethyl-7-aminodecephalosporanic acid is converted to the benzhydryl ester, which, when reacted with nitrite, is converted to the desired benzhydryl ester of 3-pyridinmethyl-7-diazodecephalosporanic acid.

The starting compound was prepared by reacting cephalosporin C with pyridine, followed by acid hydrolysis as described in USA 3 117 126.

Example 25

3-N- (2-Chloroethyl) -carbamoyloxymethyl-7-diazodecephalosporanoic acid benzhydryl ester

In the procedure described in Examples 2A, 3-1 N- (2-chloroethyl) -carbamoyloxymethyl-7-aminodecephalosporanic acid was converted to the benzhydryl ester, which was then diazotized according to the procedures described in example 2B to give the desired diazocephalosporant.

Substituted aminodecephalosporanic acid used as starting material in this example can be obtained by subjecting 7 tert -butyloxycarboxy-7-ACA to enzymatic hydrolysis to give the desired compound which is reacted with β-chloroethylisocyanate in dimethylformamide to give dimethylformamide -14- (2-chloroethyl) -carbamoyloxymethyldecephalosporin, which is hydrolyzed to give 3-14- (2-chloroethyl) -carbamoyloxymethyl-7-aminodecephalosporic acid.

Example 26

3-hydroxymethyl-7-diazodecephalosporanic acid, lactone

A mixture of sodium nitrite 10 g, 4 g lactone of 3-hydroxymethyl-7-aminodecephalosporic acid, 300 ml methylene chloride and 300 ml water and ice was shaken in a separatory funnel. P-toluenesulfonic acid monohydrate (1.6 g) was added in 3 portions for 20 min. The reaction mixture was shaken vigorously during the addition of sulfonic acid. The methylene chloride layer was separated, dried over sodium sulfate and evaporated under reduced pressure to give the desired lactone of 3hydroxymethyl-7-diazodecephalosporanic acid. The lactone of 3-hydroxy methyl-7-am and nodecephalosporanic acid is obtained by hydrolysis of cephalosporin C by known procedures.

Example 28

3-n-Amyloxymethyl-7-diazodecephalosporanoic acid benzhydryl ester

To a suspension of 6.8 g of 3-n-amyloxymethyl-7-aminodiazodecephalosporanic acid in 300 ml of dioxane at room temperature was added 4.3 g of p-toluenesulfonic acid monohydrate with stirring. The resulting solution was concentrated under reduced pressure and washed twice with dioxane. The residue was dissolved in 300 ml of dioxane at room temperature and a solution of 10 g of diphenyldiazomethane in 25 ml of dioxane was added dropwise over 15 minutes. The solution was stirred for an additional 30 min and then 25 ml of methanol was added to destroy the excess diphenyldiazomethane. The mixture was concentrated under reduced pressure and the residue partitioned between 200 ml of methylene chloride and 200 ml of water containing 10 g of dicalium sulfate, pH 8.5. The organic phase was washed with water, dried over sodium sulfate and concentrated under reduced pressure to give the desired 3-n-amyloxymethyl-7-aminodecephalosporanic acid benzhydryl ester. To a stirred mixture of 1.6 g of sodium nitrite, 30 ml of water and 40 ml of methylene chloride at 0 ° C was added 0.8 g of the ester obtained above and then a solution of 0.8 g of p-toluenesulfonic acid was added. in 5 ml of water for several minutes. The mixture was stirred at 0 ° C for 20 min and the organic phase separated, washed with a small amount of ice water, dried over anhydrous sodium sulfate at 0 ° C, filtered and concentrated under reduced pressure at room temperature. yields 3-n-amyloxymethyl-7-diazodecephalosporanoic acid benzhydryl ester.

Examples 29 - 32 3-Methyl-7-diazodecephalosporanoic acid p-methoxybenzyl ester obtained in Example 19 D, 3-benzoylthiomethyl-7 diazodecephalosporanoic acid phenyl ester obtained in Example 21D, 3-picolinoyl di thiolodiophosphoic acid 7 Example of Example 23, 3-Pyridinmethyl-7-diazodecephalosporanoic acid benzhydryl ester of Example 24, 3-14- (2-Chloroethyl) carbamoyloxymethyl-7-diazodecephalosporanoic acid benzhydryl ester of Example 25 and 3-n-methyl-methyloxymethyl ester 7-diazodecephalosporanic acid by

example 28 interact every- the product was isolated following the procedure described in Example 2C. The following products are obtained: separately with bromide azide, trimethylammonium azide and the resulting reaction Example Product No 29 3-methyl- p-methoxybenzyl ester 7-Azido-7-bromo-decaleposporic acid 30 3-Benzoylthiomethyl-7-azido-7-bromo-decephalosporic acid phenacyl ester 31 3-picolinoylthio- benzyl ester methyl-7-azido-7-bromodecephalospor new acid 32 3-Pyridinmethyl-7-azido-7-bromo-decephalosporanoic acid benzhydryl ester

Example 33

A. 3-Methyl-7-azido-7-chlordecephalosporanic acid p-methoxybenzyl ester The p-methoxybenzyl ester p-methoxybenzyl ester (0.9 g) was dissolved in a mixture of 10 ml of methylene chloride and 10 ml of nitromethane. To this cooled solution at 0-10 ° C was added a solution of triethylammonium azide, obtained as described in Example 2C, and methylene chloride solution of chlorazide (0.31 N, 15 ml) and then 50 ml of water. The resulting reaction mixture was adjusted to pH 8 by the addition of solid sodium bicarbonate. The mixture was allowed to stand and the organic layer was separated, extracted with 2x20 ml of water, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give p-methoxybenzyl ester.

3-methyl-7-azido-7-chlordecephalosporanic acid.

B. When an equivalent amount of 7-diazocephalosporanic acid benzyl ester, 7-diazocephalosporic acid benzhydric ester, 3benzoylthiomethyl-7-diazocephalosporic acid 3-benzoylthiospilosinobenzyloxyphenyl ester, 3-carbamoyloxymethoxymethoxymethoxymethoxymethoxymethyl methoxymethyl ether -7-diazodecephalosporanic acid, 3-piperidinylmethyl-7-diazodecephalosporanoic acid benzhydryl ester, benzhydryl ester

3-N- (2-Chloroethyl) carbamoyloxymethyl-7-diazodecephalosporanoic acid, 7-dia35 zocephalosporanic acid tert-butyl ester or 3-n-amyloxymethyl-7-diazodecephalosporanoic acid benzyl ester is substituted for p-methoxybenzyl esterbenzyloxy -methyl-740 diazocephalosporanic acid according to the method of Example 3A, the following compounds are obtained: 7-azido-7-chlorocephalosporanic acid benzyl ester; 7-Azido-7-chlorocephalosporanoic acid benzhydryl 45 ester; 3-benzoylthiomethyl-7-azido-750 chlordecephalosporanic acid phenacyl ester; 3-Carbamoyloxymethyl-7-chlordecephalosporanoic acid trimethylsilyl ester; benzyl ester of

3-picolinylthiomethyl-7-azido-7-chlorodecephalosporic acid; 3-Pyridinmethyl-7-azido-7-chlordecephalosporanoic acid benzhydryl ester; 3-N- (2-chloroethyl) carbamoyloxymethyl-7-azido-7-chlorodecephalosporanoic acid benzhydryl ester; 7-Azide-7-chlorocephalosporanic acid tert-butyl ester; 3-Nyloxymethyl-7-azido-7-chlordecephalosporanoic acid benzhydryl ester.

Example 34

A. 7-Azido7- (2-bromethoxy) cephalosporic acid benzhydryl ester

A solution of 543 mg of 7-azido-7-bromocephalosporanic acid benzhydryl ester in 25 ml of dry acetonitrile was treated with 195 mg of silver fluoroborate and 150 mg of 2-bromoethanol. The mixture was stirred for 24 h at room temperature, filtered and evaporated at 30 ° C (0.1 mm). The residue was chromatographed on silica gel eluting with chloroform to give the pure 7-azido-7- (2-bromethoxy) cephalosporic acid benzhydryl ester,

B. 7- (2-Thienylacetamido) -7- (2-bromethoxy) cephalosporanic acid benzhydryl ester

7-Azido-7 (2-bromethoxy) cephalosporant benzhydryl ester (587 mg) was hydrogenated at 40.6,895 kN / m ² for 3 h using 1g of platinum oxide in 50 ml of dioxane containing 500 mg of thienylacetic anhydride . After filtering the catalyst, the solution was treated for 15 min with 0.5 ml of water to hydrolyze the excess anhydride, and then evaporated in vacuo. The crude product was stirred in 25 ml of ethyl acetate, washed with aqueous sodium bicarbonate solution, dried over magnesium sulfate, filtered, evaporated and chromatographed on silica gel using 4: 1 chloroform: ethyl acetate to elute pure 7- (2-Thienylacetamido) -7- (2-bromethoxy) cephalosporic acid benzhydryl ester.

C. 7- (2-Thienylacetamido) -7- (2-triphenylphosphonoxy) cephalosporic acid benzhydryl ester, bromide

7- (2-Thienylacetamido) -7- (2-bromethoxy) cephalosporanic acid benzhydryl ester (673 mg) was refluxed in 15 ml of benzene with 262 mg of triphenylphosphine overnight. 7- (2-Thienylacetamido) -7- (2-triphenylphosphethoxy) cephalosporanic acid, bromide, is a crude benzhydryl ester obtained by evaporation of benzene and used directly for the next step.

D. 7- (2-Thienylacetamido) -7-hydroxycephalosporanic acid benzhydryl ester

Sample of 7- (2-trienylacetamido) -7- (2-triphenylphosphethoxy) cephalosporic acid benzhydryl ester, bromide obtained in the above step, about 1 mmol, was stirred under nitrogen at -30 ° C in 50 ml of ether . An equivalent amount of phenyllithium in ether was added slowly over 30 minutes. The reaction mixture was allowed to warm to 0 ° C with stirring for about half an hour. The reaction mixture was mixed with 50 ml of 0.1 M aqueous phosphate buffer at pH 7 at 0 ° C. The ether layer was separated and the aqueous phase was washed twice more with ether. The combined organic portions were dried over magnesium sulphate, filtered, evaporated and chromatographed on silica gel. It was eluted with 3% methanol in chloroform to give 7- (2-thienylacetamido) -7-hydroxycephalosporanic acid benzhydryl ester after evaporation of the solvent under reduced pressure.

F. 7- (2-Thienylacetamido) -7-hydroxy cefosulfuric acid sodium salt of acid

7- (2-Thienylacetamido) -7- (2-thienylacetamido) -7hydroxycephalosporanic acid benzhydryl ester (566 mg) was dissolved in 0.8 g of anisole and cooled to 0 ° C. Trifluoroacetic acid (4 ml), pre-cooled to 0 ° C, was added to the mixture and the reaction was allowed to continue at 0 ° C for 2 min. Immediately thereafter a 0.1 mm vacuum was applied and the reaction mixture was allowed to warm to room temperature without external heating. The anisole was then distilled at 30 ° C / 0.1 mm. A few ml of anisole were added to the residue and evaporated at 30 ° C / 1 mm to ensure complete removal of the trifluoroacetic acid. The product was treated with 5 ml of water containing 84 mg of sodium bicarbonate and lyophilized. The resulting powder was washed thoroughly with ether and dried to give 7- (2-thienylacetamido) -7-hydroxycephalosporanic acid sodium salt.

Examples 35-39

When Example 34, step B, was performed using phenylacetic anhydride, acetic anhydride, benzothiophene-2-acetic anhydride, furylacetic anhydride or tetraazolylacetic anhydride instead of thienylacetic anhydride, the corresponding 2-benzidyl esteramide was obtained (7-benzidyl esteramide). bromethoxy) cephalosporanic acid. These intermediates, when processed according to the procedures described in C, D and E, give respectively:

Example No. Product 35 7-Benzylamido-7-hydroxycephalosporanic acid sodium salt 36 7-Acetamido-7-hydroxycephalosporanic acid sodium salt 37 7- (2-Thianaphthene-2acetamido) -7-hydroxycephalosporanic acid sodium salt 38 7- (2-Furylacetamido) -7-hydroxycephalosporanic acid sodium salt 39 7-Tetrazolylacetamido-7-hydroxycephalosporanic acid sodium salt

Example 40

A. 7- (2-Thienylacetamido) -7- (aminocarbonyloxy) cephalosporanic acid benzhydryl ester

7- (2-Thiye 45 nilacetamido) -7-hydroxy cephalosporanic acid benzhydryl ester (556 mg) was dissolved in 30 ml of ether and treated at 0 ° C with 80 mg of aminocarbonyl chloride and 0.3 ml of pyridine. After 15 min, the solution was washed successively with water, dilute aqueous phosphoric acid, buffered to pH 2, water, aqueous sodium bicarbonate solution. After drying over magnesium sulfate, filtration and evaporation of the solvent, the residue is essentially pure 7- (2-thienylacetamido) -7-aminocarbonyloxy) cephalosporic acid benzhydryl ester.

B. 7- (2-Thienylacetamido) -7-aminocarbonyloxy) cephalosporanic acid sodium salt

7- (2-Thienylacetamido) -7- (aminocarbonyloxy) cephalosporanic acid benzhydryl ester was released by the procedure described in Example 34 E above to give the 7- (2-thienylacetamido) 7-aminocarbonyloxy) cephalosporanoic sodium salt. acid.

Example 41

A. 7- (2-Thienylacetamido) -7- (methoxycarbonyloxy) cephalosporanic acid anhydrous ester

To the 7- (2-thienylacetamido) -7-hydroxycephalosporanic acid benzhydryl ester in 30 ml of ether was added 95 mg of methylchlorocarbonate and 0.3 ml of pyridine. After 15 min, the reaction mixture was worked up as described in Example 40A. 7- (2-Thienylacetamido) -7- (methoxycarbonyloxy) cephalosporanic acid benzhydrol ester is obtained.

B. 7- (2-Thienylacetamido) -7- (methoxycarbonyloxy) cephalosporanic acid sodium salt

7- (2-Thienylacetamido) -7- (methoxycarbonyloxy) cephalosporanic acid benzhydryl ester was unblocked according to the procedure described in Example 34 E to give 7 (2-thienylacetamido) -7- (methoxycarbonyloxy) cephalosporanic acid sodium salt.

Example 42

A. 7- (2-Thienylacetamido) -7- (aminosulfonyloxy) cephalosporic acid benzhydrol ester

To the 7- (2-thienylacetamido) -7-hydroxycephalosporanic acid benzhydryl ester in 30 ml of ether was added 116 mg of aminosulfonyl chloride and 0.3 ml of pyridine. After 15 min, the reaction mixture was treated as described in Example 40A to afford 7- (2-thienylacetamido) -7- (aminosulfonyloxy) cephalosporanic acid benzhydryl ester.

B. 7- (2-Thienylacetamido) -7- (aminosulfonyloxy) cephalosporic acid sodium salt

7- (2-Thienyl cetate m and k) - 7- (and m-nosulfonyl) cephalosporanic acid benzhydryl ester was released according to the procedure described in Example 34 E. The sodium salt of 7- (2-th and l and cetate m ido) - 7 - (am and nos lfon and loxi) cephalosporanic acid.

Example 43

A. 7- (p-Nitrobenzylidenamino) cephalosporanic acid-benzhydryl ester

7-Aminocephalosporanic acid benzhydryl ester 438 mg was refluxed for 1 h in 50 ml of benzene with 151 mg of p-nitrobenzaldehyde in an azeotropic drying apparatus. The solvent was distilled off in vacuo to leave 571 mg of crystalline product. If desired, it can be recrystallized from benzene cyclohexane (1: 2) to give the product pure.

B. 7- (p-Nitrobenzylidenamino) - 7-cetoxy cephalosporanic acid benzhydrol ester

7- (p-Nitrobenzylidenamino) cephalosporanic acid benzhydryl ester (571 mg) was stirred at 0 ° C under nitrogen in 10 ml of acetonitrile. Acetyl peroxide (118 mg) was added followed by 129 mg of diisopropylethylamine in acetonitrile over a period of 5 min.

The reaction mixture was stirred at room temperature for 5 min and evaporated under reduced pressure. The residue was stirred in 25 ml of benzene and washed sequentially with water, dilute phosphoric acid, buffered at pH 2, water and aqueous sodium bicarbonate solution. The solution was dried over magnesium sulfate, filtered, evaporated and chromatographed on silica gel and 4: 1 chloroform-ethyl acetite was used to elute the product. Evaporation of the solvent gave 7- (p-nitrobenzylidenamino) -7-acetoxycephalosporanic acid benzhydryl ester.

C. 7-amino-7-acetoxycephalosporanic acid benzhydryl ester, hydrochloride

Benzhydryl 7- (p-nitrobenzylidenamino) -7-acetoxycephalosporanate 629 mg and 260 mg aniline hydrochloride were stirred together for 1 h at 25 ° C in 10 ml of methanol. The methanol was removed at 0.1 mm pressure and 30 ° C and the residue was coated with ester to crystallize for 1 h. The solid was triturated with ether, the solid was filtered and washed several times with ether. 7-Amino-7-acetoxycephalosporic acid benzhydryl ester, hydrochloride and aniline, combined together are used, which are used immediately in the next step.

D. 7- (2-Thienylacetamido) -7-acetoxycephalosporanoic acid benzhydryl ester

A mixture of 7amino-7-acetoxycephalosporanic acid benzhydryl ester, hydrochloride and analinhydrochloride obtained in the previous step was stirred vigorously at -10 ° C in 25 ml of methylene chloride. 0.5 g of thienylacetyl chloride is added, followed by slowly adding 0.5 g of triethylamine.

A free 7-amino compound is obtained which is acetylated immediately. The reaction mixture was slowly allowed to warm to room temperature. The excess acid chloride is hydrolyzed by shaking with water and the methylene chloride layer is then washed sequentially with dilute phosphoric acid, buffered to pH 2, water and dilute aqueous sodium bicarbonate solution. After drying over magnesium sulfate, the solution was filtered and evaporated to give a mixture of product and N-phenylthienylacetamide. Chromatography on silica gel, eluting with 4: 1 chloroform ethyl acetate gave the title compound.

E. 7- (2-Thienylacetamido) -7-acetoxycephalosporanic acid sodium salt

7- (2-Thien and l-acetam ido) -7-cetoxy cerebro losporanic acid benzhydryl ester (620 mg) was dissolved in 0.8 ml of anisole and cooled to 0 ° C. Pre-cooled to 0 ° C trifluoroacetic acid (4ml) was added and the reaction was allowed to proceed at 0 ° C for 2 min. A vacuum of 0.1 mm is then applied immediately and the reaction mixture is allowed to warm to room temperature. The anisole was then distilled at 30 ° C 0.1mm. Add a few ml of anisole to the residue and evaporate again at 30 ° C / 0.1 mm to ensure complete elimination of the trifluoroacetic acid. The product was treated with 5 ml of water containing 84 mg of sodium bicarbonate and lyophilized. The resulting powder was washed thoroughly with ether and dried to give the title compound.

Example 44

A. 7-azido-7-bromcephalosporanic acid p-methoxybenzyl ester

Mixture of p-methoxybenzyl ester of 7-aminephalosporanic acid

3.9 g in 250 ml of methylene chloride and sodium nitrite 1.4 g in 250 ml of water was stirred under ice with ice-cooling and 7.5 ml of 2N sulfuric acid was added repeatedly. The mixture was stirred at 3 ° C for one hour and the organic phase was separated and dried rapidly over sodium sulfate and concentrated to 50 ml in vacuo. To this solution of p-methoxybenzyl ester of 7-diazocephalosporanic acid, cooled to -40 ° C, was added rapidly 50 ml of triethylammonium azide solution and 50 ml of bromazide solution prepared as described below. The solution was allowed to warm to 0 ° C, then washed with 200 ml of 2% sodium hydrogen phosphate solution. The organic phase was dried over sodium sulfate and evaporated. The residue was chromatographed on a silica gel column 4.5 cm x 15 cm using 20% methylene chloride 80% benzene as eluent. 7-Azido7-bromocephalosporanic acid p-methoxybenzyl ester is obtained.

Preparation of triethylammonium azide solution. To a suspension of 6 g of sodium azide in 10 ml of water and 100 ml of methylene chloride at -10 ° C was added 10 ml of 50% sulfuric acid. The organic phase was decanted from the aqueous paste and dried over anhydrous magnesium sulfate. 2.25 ml of triethylamine are added to 50 ml of the hydrochloric acid solution. The pH obtained is 7.

Preparation of bromazide solution. To 50 ml of the above hydrochloric acid solution was added 2 g of N-bromosuccinamide with stirring and ice cooling. After 10 min at 0 ° C the reaction was complete and the solution was used immediately for reaction with 7-diazocephalosporanate.

7-Nocephalosporanic acid p-mitoxybenzyl ester used as starting material was prepared from 7-aminocephalosporanic acid using the procedure described in Example 19A, B and C to obtain 3-methyl- p-methoxybenzyl ester. 7-Aminocephalosporanic acid.

B. 7-azido-7- (2-methoxyethoxy) -cephalosporanic acid p-methoxybenzyl ester

To a solution of 4.8 g of p-methoxybenzyl ester of 7-azido-7-bromocephalosporanic acid in 20 ml of 2-methoxyethanol was added 1.95 g of silver tetrafluoroborate and 0.8 ml of pyridine. After stirring for 3 h at room temperature, the 2-methoxyethanol was removed in vacuo. The residue was taken up in methylene chloride, filtered and the filtrate washed with water, dried and evaporated. The residue was chromatographed on 140 g of silica gel. Elution with 2% methanol in chloroform afforded pure 7-azido-7- (2-methoxyethoxy) cephalosporic acid p-methoxybenzyl ester.

C. amino-7- (2-methoxyethoxy) -cephalosporanic acid p-methoxybenzyl ester p-7-azido-7-12-methoxyethoxy) -cephalosporic acid p-methoxybenzyl ester (2 g) and platinum oxide (2g) in 200 ml The dioxane was stirred vigorously under nitrogen for one hour. The fresh catalyst (2g) was added and the hydrogenation continued for 2 hours. The dioxane was evaporated under reduced pressure and the catalyst was absorbed on 30 g of silica gel in 50 ml of chloroform and filtered. The filter cake was washed with 500 ml of chloroform and the combined filtrate and washings were evaporated. 7-Amino7- (2-methoxyethoxy) -cephalosporanic acid p-methoxybenzyl ester was obtained as a viscous oil.

D. (O-a-Azidophenylacetamido) -7- (2-methoxyethoxy) -cephalosporanoic acid p-methoxybenzoyl ester

To an ice-cooled solution of 0.9 g of p-methoxybenzene ester of amino-7- (2-methoxyethoxy) -cephalosporanic acid in 20 ml of methylene chloride was added 0.8 g of D-a-azidophenylacetyl chloride and 0.7 ml of dry pyridine. The solution was stirred at 2 ° C for 20 min, then poured into 20 ml of water. The mixture was acidified to pH 2 with dilute sulfuric acid and the organic phase was separated, washed with water, dried and evaporated. The residue was chromatographed on 50 g of silica gel. Elution with chloroform-ethyl acetate affords N- (D-a-azidophenylacetamido) -7 (2-methoxyethoxy) -cephalosporanic acid p-methoxybenzyl ester.

E. 7- (O-a-azodiphenylacetamido) 7- (2-methoxyethoxy) -cephalosporic acid

A solution of 420 mg of 7- (O-a-azidophenylacetamide) -7- (2-methoxyethoxy) -cephalosporic acid p-methoxybenzyl ester, 300 mg of anisole and 3 ml of trifluoroacetic acid in 10 ml of benzene was stirred for 2 h at room temperature. The reaction mixture was evaporated and the residue was taken up in ethyl acetate and extracted with sodium bicarbonate solution. The aqueous extract was cooled to 0 ° C, covered with ethyl acetate and acidified to pH 2.1 with dilute sulfuric acid. The ethyl acetate layer was separated, dried over anhydrous sodium sulfate and evaporated to give 7- (O-a-azidophenylacetamide) -7- (2-methoxyethoxy) -cephalosporic acid.

F. 7- (D-a-Aminophenylacetamide) 7- (2-methoxyethoxy) -cephalosporic acid

To a ice-cold solution of 240 mg of 7- (0-a-azidophenylacetamide) -7- (2-methoxyethoxy) -cephalosporanic acid in 3 ml of acetic acid and 2 ml of water was added 1.5 g of zinc powder. The mixture was stirred for 15 min after being filtered. The residue was washed with 10 ml of water and the combined filtrate and washes were evaporated to dryness. The residue was mixed with 2 ml of water and poured into a 2 x 40 cm ion retaining column. The column was washed with water and ninhydrin, the positive fractions were lyophilized to give 7- (D-? -Aminophenylacetamide) -7- (2-methoxyethoxy) -cephalosporanic acid as a yellow powder.

Example 45

A. 7 (2-Methoxyethoxy) -7- (2-thienylacetamide) -cephalosporic acid p-methoxybenzyl ester

In the procedure described in 3B, the 7-amino-7- (2-methoxyethoxy) -cephalosporanoic acid p-methoxybenzyl ester, equivalent to a quantitative amount, was reacted with 2-thienyl chloride and the reaction product obtained was treated in the same manner. 7- (2-Methoxyethoxy) -7- (2-thienylacetamide) -cephalosporanic acid p-methoxybenzyl ester is obtained.

B. 7- (2-Methoxyethoxy) -N- (2-thienylacetamide) -cephalosporanoic acid sodium salt

A solution of 0.4 g of 7- (2-methoxyethoxy) -7 (2-thienylacetamido) -cephalosporanic acid p-methoxybenzyl ester was dissolved in 3.5 ml of anisole and treated with 10 ml of trifluoroacetic acid at room temperature for 24 hours. 10 min. Trifluoroacetic acid and anisole were removed under reduced pressure while maintaining the temperature below 40 ° C and the residue was taken up in 25 ml of chloroform and treated with 20 ml of water containing 0.120 g of sodium bicarbonate. The mixture was stirred for half an hour at room temperature and the organic phase was separated and washed with water. The combined aqueous phase was washed twice with methylene chloride and lyophilized to give the 7- 5 (2-methoxyethoxy) -7- (2-thienylacetamido) cephalosporanic acid sodium salt.

Examples 46 to 50

In the manner described in Example 45, the amino-7- (2-methoxyethoxy) cephalosporanoic acid p-methoxybenzyl ester is reacted with 2thyanaphthen-2-acetyl chloride, furylacetyl chloride, phenylacetyl chloride or tetrazolylacetyl chloride and the resulting tetrazolylacetyl chloride. The following products are obtained:

Example No. Product 46 7- (2-Methoxyethoxy) sodium salt - 7- (2-cyanophen-2-acetamide) -cephalos- - tartaric acid 47 7- (2-methoxyethoxy) -7- (p-guanidinephenylacetamide) -cephalosporic acid 48 7- (2-Methoxyethoxy) sodium salt - 7- (2-furylacetamide) -cephalosporanic acid 49 7- (2-Methoxyethoxy) 7-phenylacetamidcephalosporanic acid sodium salt 50 7- (2-Methoxyethoxy) sodium salt - 7-Tetrazolylacetamide-cephalosporic acid

Example 51

A. 7-Azide7 (tert-butoxycarbonylaminoacetoxy) -cephalosporanic acid benzhydryl ester

A mixture of 3.1 g of 7-azido-7-bromcephalosporanic acid benzhydryl ester, 1.1 g of silver fluoroborate, 2 g of t-butoxycarbonylglycine and 0.45 ml of pyridine in 10 ml of dioxane are stirred in the absence of moisture and light 3 hours at room temperature. The mixture was diluted with 100 ml of ether and filtered, the filtrate washed successively with 2% aqueous phosphoric acid, water and 5% aqueous sodium bicarbonate solution. The ether phase was dried over anhydrous sodium sulfate and evaporated and the residue chromatographed on 140 g of silica gel. Elution with 2% methanol in chloroform afforded 7-azide-7- (tert-butoxycarbonylaminoacetoxy) -cephalosporanic acid benzhydryl ester.

B. 7-Amino-7 (-tert.-butoxycarbonylaminoacetoxy) -cephalosporanic acid benzhydryl ester

A solution of 3 g of benzhydryl ester of 7-azido-7-tert. (-butoxycarbonylaminoacetoxy) -cephalosporanic acid in 300 ml of dry dioxane is hydrogenated in the presence of 3 g of platinum oxide for 1 h at room temperature. 3 g of fresh catalyst was added and the hydrogenation continued for 3 h. The dioxane was evaporated under reduced pressure and the residue was mixed with chloro57 form and filtered through 20 g of silica gel, placed in a porous bottom funnel. The filter cake was washed with 500 ml of chloroform and the combined washes and the filtrate evaporated to give benzhydryl 7-amino-7 (tert-butoxycarbonylaminoacetoxy) -cephalosporic acid.

C. 7- (tert, butoxycarbonylaminoacetoxy) -7- (2-furylacetamide) -cephalosporanoic acid benzhydryl ester

To a cooled solution of 2 g of 7-amino- (tert-butoxycarbonylaminoacetoxy) -cephalosporanoic acid benzhydryl ester was added 1.5 ml of 2-furylacetyl chloride and 1.5 ml of dry pyridine. The mixture was stirred at 0 ° C for 15 min and then poured onto ice. The methylene chloride layer was separated, washed sequentially with 2% phosphoric acid, water and 2% sodium bicarbonate solution, dried and evaporated. The residue was chromatographed on 80 g of silica gel. It was eluted with a mixture of chloroformethyl acetate. Essentially pure benzhydryl 7- (tert-butoxycarbonylaminoacetoxy) -7- (2-furylacetamide) -cephalosporanic acid is obtained.

D. 7-aminoacetoxy-7- (2-furylacetamide) -cephalosporanoic acid, trifluoroacetic acid salt

A solution of 400 g of benzhydryl 7 (tert-butoxycarbonylaminoacetoxy) -7- (2-furylacetamide) -cephalosporanic acid in 3 ml of anisole and 3 ml of trifluoroacetic acid was kept at room temperature for 5 min, after which the excess of trifluoroacetic acid was reduced vacuum for 5 min. The residue was triturated with ether and the resulting powder was filtered off and washed with ether. The residue was dissolved in 200 ml of water and filtered through 50 g of charcoal coated on diatomaceous earth. The filtrate was lyophilized to give 7-aminoacetoxy-7- (2-furylacetamide) -cephalosporanic acid, a trifluoroacetic acid salt as a light powder. The internal ammonia salt is obtained by neutralizing the aqueous solution to pH 6.5, followed by concentration to a small volume and precipitation with alcohol.

Example 52

By repeating the process of Example 51 and using tetraazolylacetyl chloride, 2-thienylacetyl chloride or phenylacetyl chloride instead of 2-furylacetyl chloride, the corresponding 7-tetraazolylacetamide, 7- (2-thienylacetamide) or 7-phenylacetylacetate7, triacetylacetate,

Pimer 53

A. 7-Azido7-benzyloxycephalosporanoic acid benzhydryl ester

A mixture of 4.6 g of 7-azido-7-bromcephalosporanic acid benzhydryl ester, 1.6 g of silver fluoroborate and 0.66 ml of dry pyridine in 15 ml of benzyl alcohol was stirred at room temperature for 2 1/2 hours. The mixture was diluted with 300 ml of ether and filtered. The filtrate was washed with 2% phosphoric acid and water, then dried over sodium sulfate and evaporated. The excess benzyl alcohol was removed on a high vacuum magnetic stirring pump for 18 h and the residue was chromatographed on 200 g of silica gel. It was eluted with hexane, then with methylene chloride in hexane with increasing concentration. 7-Azido-7-benzyloxycephalosporanoic acid benzhydryl ester is obtained

B. 7-amino-7-benzyloxycephalosporanic acid benzhydryl ester

A solution of 3 g of 7-azido-7-benzyloxycephalosporanic acid benzhydryl ester in 300 ml of dry dioxane was hydrogenated in the presence of 3 g of platinum oxide at room temperature and atmospheric pressure for 1 h. Fresh catalyst (3 g) was added and hydrogenation continued. 2 h. The dioxane was removed under reduced pressure and the residue was stirred with chloroform and filtered through 20 g of silica gel placed in a foamed funnel. The filter cake was washed with 500 ml of chloroform and the combined filtrate and washes were evaporated. 7-Amino-7-benzyloxycephalosporanic acid benzhydryl ester was obtained as a yellow gum.

C. 7-Benzyloxy-7- (2-thienylacetamide) -cephalosporanic acid benzhydryl ester

To a cooled solution of 1.2 g of 7-amino-7-benzyloxycephalosporic acid benzhydrol ester in 15 ml of methylene chloride was added 0.8 ml of 2-thienylacetyl chloride and 0.8 ml of pyridine with stirring. The mixture was stirred for 15 min at 2 ° C and then poured onto ice. The organic phase was separated and washed sequentially with 2% phosphoric acid, water and 2% sodium bicarbonate solution, then dried over anhydrous sodium sulfate and evaporated. The residue was chromatographed on 50 g of silica gel with 5% ethyl acetate in methylene chloride as eluent to give 7-benzyloxy-7- (2-thienylacetamide) -cephalosporic acid benzhydryl ester.

D. 7-Benzyloxy7- (2-thienylacetamide) -cephalosporanic acid sodium salt

A solution of 0.9 g of 7-benzyloxy-7- (2-thienylacetamide) -cephalosporanic acid benzhydryl ester in 3 ml of anisole and 7 ml of trifluoroacetic acid was stirred at room temperature for 5 min and the excess anisole and trifluoroacetic acid were rapidly removed. vacuum pump. The residue was stirred with a mixture of 0.25 ml of methylene chloride and 25 ml of water and stirred rapidly until sodium bicarbonate was added to adjust the pH to 6.5. The aqueous layer was separated, washed once with ether and lyophilized. 7-Benzyloxy-7- (2-thienylacetamide) -cephalosporanic acid sodium salt was obtained as a light powder.

Example 54

7-Hydroxy-7- (2-thienylacetamide) -cephalosporanic acid sodium salt

A solution of 7-benzyloxy-7- (2-thienylacetamide) -cephalosporanic acid sodium salt (300 mg) in 10 ml of water was hydrogenated at 40.6,865 kN / m ² and room temperature in the presence of 0.39 g of 10% palladium on carbon for 6 h. 0.3 g of fresh catalyst was added and the hydrogenation continued for 18 h. The catalyst was filtered off and the filtrate was lyophilized to give 7-hydroxy-7- (2-thienylacetamido-cephalosporanic acid sodium salt).

Example 55

Following the procedure described in Example 53 and using furylacetyl chloride, p-guanidinephenylacetyl chloride hydrochloride, tetrazolylacetyl chloride or phenacetyl chloride instead

2-thienylacetyl chloride to give, respectively: 7-benzyloxy-7- (2-furylacetylamide) -cephalosporanoic acid sodium salt, 7-benzyloxy7- (p-guanidinephenylacetamide) -cephalosporanoic acid, 7-benzytosiloxy-7-benzyloxy-acetylacetylacetylacetate 7-Benzyloxy-7-phenylacetamidcephalosporic acid acid and sodium salt

Example 56

In the procedure described in Example 54, 7-benzioxycephalosporins are converted to the corresponding 7-hydroxy59 cephalosporins.

Example 57

A. 7-Azido7- (B-2-benzhydryloxycarbonyl-2-tert-butoxycarbonylaminoethoxy) cephalosporic acid benzhydryl ester

N-tert-butoxycarbonyl-L-serine benzhydrol ester was obtained by adding a solution of diphenyldiazomethane (2.5g) in 10 ml dioxane to a solution of N-tert-butoxycarbonyl-B-serine (2.5 g) in 5 ml of dioxane. To the resulting solution was added 7-azido-7-bromo-cephalosporanic acid benzhydryl ester 4.5 g, silver fluoroborate 1.6 g and pyridine 0.65 ml. The mixture was stirred for 3 h at room temperature, then diluted with 100 ml of ether and filtered. The filtrate was washed sequentially with phosphoric acid buffer with pH 2, water, sodium bicarbonate solution and then dried over anhydrous sodium sulfate and evaporated. The residue was chromatographed on 200 g of silica gel. Elution with chloroform-ethyl acetate gave the product as a tan.

B. 7-Amino-7- (L-2-benzhydryloxycarbonyl-2t-butoxycarbonylaminoethoxy) cephalosporic acid benzhydryl ester

A solution of 3 g of 7-azido-7- (L-2-benzhydryloxycarbonyl-2-tert-butoxycarbonylaminoethoxy) cephalosporic acid benzhydryl ester in 300 ml of dry dioxane is hydrogenated in the presence of 3 g of platinum oxide at room temperature and atmospheric pressure. 1 h. Fresh catalyst (3 g) was added and hydrogenation continued for 2 h. Dioxane was removed under reduced pressure and the residue was stirred in chloroform and filtered through 20 g of silica gel, packed in a glass filter. The filter cake was washed with chloroform and the combined washings were evaporated to give 7-amino7- (6- (2-benzylhydroxycarbonyl-2-tert.-butoxycarbonylaminoethoxy) cephalosporic acid benzhydryl ester.

C. Benzhydryl 7- (a-benzhydryloxycarbonylphenylacetamido) -7- (B-2benzhydryloxycarbonyl-2-t-butoxycarbonylaminoethoxy) -cephalosporic acid a-benzhydryloxycarbonylphenylacetyl chloride 450 mg mgster 7- (L-2-benzhydryloxycarbonyl) 2 (tert-butoxycarbonylaminoethoxy) cephalosporic acid and 0.2 ml dry pyridine in 10 ml methylene chloride. The solution was stirred for 20 min at 0 ° C, then poured into 20 ml of pH 2 phosphoric acid buffer. The organic phase was washed with water and brine, dried over anhydrous sodium sulfate and evaporated. The residue was chromatographed on 30 g of silica gel. Elution with chloroform afforded 7- (α-benzhydryloxycarbonylphenylacetamido) -7- (L-2benzhydryloxycarbonyl-2-t-butoxycarbonylaminoethoxy) cephalosporic acid benzhydryl ester

D. 7- (B-2-Carboxy-2-aminoethoxy) -7- (α-carboxyphenylacetamido) cephalosporic acid disodium salt ml of anisole and 6 ml of trifluoroacetic acid are added to 200 (7- (α-benzhydryloxycarbonyloxy) carbonyloxycarbamoylacetylcarbamoyl ester) -7 (B-2-Benzhydryloxycarbonyl-2-tert-butoxycarbonylaminoethoxy) cephalosporanic acid and the mixture was stirred at room temperature for 8 min. The residue was stirred in 20 ml of methylene chloride and 20 ml of water and the mixture was stirred vigorously, bringing the pH to 7 by the addition of sodium bicarbonate solution. The aqueous solution was separated, washed once with ether and lyophilized. 7- (L-2-Carboxy-2-aminoethoxy) -7 (α-carboxyphenylacetamido) cephalosporanic acid disodium salt was obtained. Similarly, starting from N-tert-butoxycarbonyl-O-serine instead of N-tert-butoxycarbonyl-L-serine, 7- (O-2-carboxy-2-aminoethoxy) -7 (a-carboxyphenylacetamido) is obtained ) cephalosporanic acid.

Example 58

When Ίamino-7- (L-2-benzhydryloxycarbonyl-2-t-butoxycarbonylaminoethoxy) cephalosporanic acid benzhydryl ester is acylated with phenylacetyl chloride, 2-thienylacetyl chloride, or the furylacetyl chloride described above according to the procedures described in the above 7 , 7- (2-thienylacetamido) and 7- (2-furylacetamido) analogs.

Example 59

7-Azido-7- (carbamoylmethoxy) cephalosporic acid

To a solution of 3.1 g of 7-azido-7-bromocephalosporanic acid benzhydryl ester and 2 g of glycolamide in 10 ml of dioxane were added 1.1 g of silver fluoroborate and 0.45 ml of pyridine. The mixture was stirred for 2 h at room temperature, then diluted with 100 ml of ether and filtered. The filtrate was washed successively with 2% aqueous phosphoric acid, water and sodium bicarbonate solution, then dried over anhydrous sodium sulfate and evaporated. The residue was chromatographed on 120 g of silica gel. Elution with 2% methanol in chloroform afforded essentially pure 7-azido-7- (carbamoylmethoxy) cephalosporic acid benzhydryl ester.

B. 7-amino-7- (carbamoylmethoxy) cephalosporanic acid benzhydryl ester

A solution of 3 g of 7-azido-7- (carbamoylmethoxy) cephalosporanic acid benzhydrol ester in 300 ml of dry dioxane was stirred vigorously with 3 g of platinum oxide under one atmosphere of hydrogen at room temperature for 1 hour. The dioxane was removed under reduced pressure. and the residue was taken up in chloroform and filtered through 20 g of silica gel packed into a glass shot filter. The silica gel was washed with chloroform and the combined filtrate was evaporated, leaving 7-amino-7- (carbamoylmethoxy) cephalosporic acid benzhydryl ester.

C. 7-Carbamoylmethoxy-7- (2-thienylacetamido) cephalosporic acid benzhydryl ester

To a cooled solution of 1.5 g of 7-amino-7- (carbamoylmethoxy) cephalosporanic acid benzhydryl ester in 20 ml of methylene chloride was added 1.0 g of thienylacetyl chloride and 1.0 ml of dry pyridine. The mixture was stirred for 15 min at 0-2 ° C and then poured into 100 ml of phosphoric acid buffer at pH 2. The methylene chloride phase was washed with water and sodium bicarbonate solution, dried over anhydrous sodium sulfate and evaporated. The product was chromatographed on 50 g of silica gel with 5% ethyl acetate-chloroform as eluent to give 7-carbamoylmethoxy-7- (2-thienylacetamido) cephalosporic acid benzhydryl ester.

D. 7-Carbamoylmethoxy-7- (2-thienylacetamido) cephalosporanic acid sodium salt

A mixture of 0.5 g of 7-carbamoylmethoxy-7- (2-thienyl61 cetamidocephalosporant benzhydryl ester in 3 ml of anisole and 7 ml of trifluoroacetic acid was stirred at 0 ° C for 10 minutes. The excess of anisole and trifluoroacetic acid was rapidly The residue was stirred in 40 ml of water and 40 ml of methylene chloride and the mixture was stirred vigorously, bringing the pH to 6.5 by the addition of sodium bicarbonate. The aqueous phase was separated, washed once with ether and lyophilized to give 7-carbamoylmethoxy-7 (2-thienylacetamido) cephalosporan sodium salt acid.

Example 60

When the when, when the when, when, when .

Example 61

A. 7-Azido7- / benzhydryloxycarbonylmethoxy / cephalosporic acid benzhydryl ester

A solution of the glycolic acid benzhydryl ester is obtained by adding a solution of 2.5 g of diphenyldiazomethane in 10 ml of dioxane to a solution of 0.7 g of glycolic acid in 5 ml of dioxane. To it is added

4.5 g of 7-azido7-bromocephalosporanic acid benzhydrol ester, 1.6 g of dry silver fluoroborate and 0.65 ml of dry pyridine. The mixture was stirred for 3 h at room temperature, then diluted with ether and filtered. The filtrate was washed with water, dried over anhydrous sulfate and evaporated. The residue was chromatographed on 160 g of silica gel to give the benzhydryl ester of

7-Azido-7- (benzhydroxycarbonylmethoxy) cephalosporic acid.

B. 7-Amino-7- (benzhydryloxycarbonylmethoxy) cephalosporic acid benzhydryl ester

A solution of 3 g of 7-azido-7- (benzhydryloxycarbonylmethoxy) cephalosporic acid benzhydryl ester in 300 ml of dry dioxane was stirred vigorously with 3 g of platinum oxide under one atmosphere of hydrogen at room temperature for 1 h. 3 g / and hydrogenation was continued for another 2 h. Dioxane was removed under reduced pressure and the residue was stirred in chloroform and filtered through 20 g of silica gel packed in a sintered glass filter. The silica gel was washed with 300 ml of chloroform and the filtrate and washing liquids were combined and evaporated, leaving 7-amino-7- (benzhydryloxycarbonylmethoxy / cephalosporic acid benzhydryl ester) as a heavy oil.

C. 7- (Benzhydryloxycarbonylmethoxy) cephalosporic acid benzhydryl ester

To a cooled solution of 0.4 g of 7-amino-7- (benzhydryloxycarbonylmethoxy) cephalosporic acid benzhydryl ester in 5 ml of methylene chloride was added 0.25 g of thienylacetyl chloride and 0.25 ml of pyridine. The mixture was stirred for 15 min at 0-3 ° C and then poured into ice. The organic phase was washed sequentially with 2% phosphoric acid, water and aqueous sodium bicarbonate solution and finally dried over anhydrous sodium sulfate and evaporated. The residue was chromatographed on 25 g of silica gel, the product was eluted with 5% ethyl acetate in methylene chloride to give 7- (benzhydryloxycarbonylmethoxy) -7- (2-thienylacetamido) cephalosporic acid benzhydryl ester.

D. 7- (Carboxymethoxy) -7- (2-thienylacetamido) cephalosporic acid disodium salt

To a solution of 0.4 g of 7- (benzhydryloxycarbonylmethoxy) -7- (2-thienylacetamido) cephalosporanoic acid benzhydryl ester in 3 ml of anisole was added 5 ml of trifluoroacetic acid. The mixture was held for 8 min, then the excess anisole and trifluoroacetic acid were rapidly evaporated in a vacuum pump. The residue was stirred in a mixture of 20 ml of methylene chloride and 20 ml of water and the pH was adjusted to 6.5 (glass electrode) with sodium bicarbonate. The aqueous phase was separated and extracted once with ether and then lyophilized to give 7- (carboxymethoxy) -7- (2-thienylacetamido) cephalosporanic acid disodium salt.

Example 62

When the 7-amino-7- / benzhydryloxycarbonylmethoxy / cephalosporanic acid benzhydryl ester is reacted with 2-cyanaphen-2-acetyl chloride or monobenzhydryl phenylmalonyl chloride instead of the thienyl chloride, the resulting reaction products are subsequently obtained as a reaction product 61 7- (2-carboxymethoxy) -7- (2-thianaphthene-2-acetamido) cephalosporic acid sodium salt and 7- (2-carboxymethoxy) -7- (2-phenylacetamido) cephalosporic acid sodium salt.

Example 63

A. 7-Acetoxy-7-azidocephalosporanoic acid benzhydryl ester

A mixture of 2.2 g of 7-azido-7-bromcephalosporanic acid benzhydryl ester and 0.8 g of silver acetate in 10 ml of acetic acid was stirred at room temperature for 3 hours. The acetic acid was removed under reduced pressure and the residue was stirred in methylene chloride and filtered. The filtrate was washed with sodium bicarbonate solution, then dried over sodium sulfate and evaporated. The residue was chromatographed on 80 g of silica gel. Elution with chloroform afforded 7-acetoxy-7-azidocephalosporanoic acid benzhydryl ester.

B. 7-amino-7-acetoxy cephalosporanic acid benzhydryl ester g 7-acetoxy-7-azidocephalosporanoic benzhydryl ester in 200 ml of dry dioxane is hydrogenated at room temperature and atmospheric pressure in the presence of 2 g of platinum

h. 2 g of fresh catalyst was added and the hydrogenation continued for 2 h. The solvent was evaporated and the residue dissolved in ether and shaken with 10 ml of anhydrous magnesium sulfate powder and filtered through an infuser soil into a glass filter. The filtrate was evaporated to leave 7-amino-7-acetoxycephalosporanic acid benzhydryl ester.

C. 7-Acetoxy-7- (2-thienylacetamido / cephalosporic acid benzhydryl ester

To a cooled solution of 1.2 g of 7-amino-7-acetoxycephalosporic acid benzhydryl ester in 20 ml of methylene chloride was added 0.8 ml of thienylacetyl chloride and 0.8 ml of pyridine. The mixture was stirred at 0 ° C for 15 min and then poured onto ice. The organic phase was washed sequentially with 2% phosphoric acid solution, water and 2% sodium bicarbonate solution, dried over anhydrous sodium sulfate and evaporated. The residue was chromatographed on 50 g of silica gel. Elution with chloroform afforded 7-acetoxy-6- (2-thienylacetamido / cephalosporanic acid) benzhydryl ester.

D. 7-2-Acetoxy7- (2-thienylacetamido / cephalosporanoic acid) sodium salt

A solution of 2 g of 7-acetoxy-7- (2-thienylacetamido / cephalosporic acid benzhydryl ester in 8ml of anisole and 16 ml of trifluoroacetic acid was stirred at room temperature for 5 min. The excess anisole and trifluoroacetic acid were rapidly evaporated and the residue was evaporated rapidly. was stirred in 50 ml of methylene chloride and extracted twice with sodium bicarbonate solution. The aqueous extract was cooled and coated with a layer of ethyl acetate and the pH was adjusted to 1.2 with dilute sulfuric acid. The ethyl acetate layer was stirred vigorously with water, bringing the pH to 6.5 (glass electrode). Evaporation of the aqueous extract afforded the 7-acetoxy-7- (2-thienylacetamido / cephalosporanic acid) sodium salt.

Example 64

3-Hydroxymethyl7-hydroxy-7- (2-thienylacetamido / decephalosporanoic acid) sodium salt

1.8 g of the 7-acetoxy-7- (2-thienylacetamido) -cephalosporanic acid sodium salt was dissolved in 50 ml of a citrus acetyl esterase solution in a water bath and heated at 30 ° C. The pH was maintained 6.6 by the addition of N sodium hydroxide solution, controlled by pH stat. After 8 h, 15 g of sodium chloride was added, the solution was covered with 100 ml of ethyl acetate and the pH was adjusted to 2.1 with hydrochloric acid. The mixture was centrifuged and the ethyl acetate supernatant removed. The ethyl acetate extract was stirred vigorously with water, bringing the pH to 7 with sodium bicarbonate solution. The aqueous phase was separated and lyophilized to give 3-hydroxymethyl-7-hydroxy-7- (2-thienylacetamido / decephalosporanoic acid sodium salt.

Example 65

3-Carbamoyloxymethyl-7-carbamoyloxy-7- (2-thienylacetamido / decephalosporic acid

To 0.4 g of 3-hydroxymethyl-7-hydroxy-7/2-thienylacetamido / decephalosporanoic acid sodium salt suspended in 10 ml of acetonitrile cooled to 0 ° C was added 0.6 ml of chlorosulfonylisocyanate. The reaction mixture was stirred for 2 h and then evaporated to dryness. The residue was stirred in a mixture of 20 ml of ethyl acetate and 20 ml of phosphate buffer and the pH was adjusted to 1.6. The mixture was stirred for 3 h at room temperature and the pH was adjusted to 8, the aqueous phase was separated. The organic phase was extracted with pH 8 phosphate buffer. The combined aqueous phase was adjusted to pH 2 with hydrochloric acid and extracted twice with ethyl acetate. The ethyl acetate extract was dried over anhydrous sodium sulfate and evaporated and the residue was washed with ether. 3-Carbamoyloxymethyl-7-carbamoyloxy-7- (2-thienylacetamido / decephalosporanoic acid is obtained as a yellow solid.

By the same procedure, using other acylating agents instead of thienylacetyl chloride, other 7-acylamido analogues of the products of Examples 63, 64 and 65 were obtained.

Example 66

A. 7-Azido7-phenoxycephalosporanic acid benzhydryl ester

To a solution of 2.3 g of 7-azido-7-bromocephalosporanic acid benzhydryl ester in 10 ml of benzene was added 6 g of phenol and then 0.82 g of dry silver fluoroborate and 0.33 ml of dry pyridine. The mixture was stirred at

23 ° C for 6 h, then filtered and the residue washed with benzene. The combined filtrate and washing liquids were washed with phosphoric acid buffer with pH 2 and aqueous sodium bicarbonate solution and then dried over sodium sulfate and evaporated. The residue was placed on a high vacuum pump overnight to remove excess phenol, then chromatographed on 100 g of silica gel. Elution with methylene chloride affords 7-azido-7-phenoxycephalosporanic acid benzhydryl ester as a viscous oil.

B. 7-amino-7-phenoxycephalosporanic acid benzhydryl ester

A solution of 3 g of 7-azido-7-phenoxycephalosporic acid benzhydryl ester in 300 ml of dry dioxane is hydrogenated in the presence of platinum oxide at room temperature and atmospheric pressure for 1 h. 3 g of fresh catalyst is added and hydrogenation is continued for 2 h The dioxane was removed under reduced pressure and the residue was stirred in chloroform and filtered through 20 g of silica gel, packed in a shot filter. The filter cake was washed with 500 ml of chloroform and the combined liquids and the filtrate evaporated, leaving 7-amino-7-phenoxycephalosporanic acid benzhydryl ester.

C. 7-Phenoxy-7- / 5-theazolylacetamido / cephalosporic acid benzhydryl ester

A solution of 2.3 g of 7-amino-7-phenoxycephalosporic acid benzhydryl ester in 25 ml of methanol was cooled in an ice bath and 1.8 ml of dry pyridine was added to it with magnetic stirring, followed immediately by a solution of

1.5 g of 5-thiazolylacetyl chloride in 4 ml of methylene chloride. The mixture was stirred at 2 ° C for 15 min, then extracted with pH 2 phosphoric acid buffer and water. The organic phase was dried over anhydrous sodium sulfate and evaporated. The residue was chromatographed on 100 g of silica gel. Elution with chloroform afforded 7-phenoxy-7- (5-thiazolylacetamido / cephalosporanic acid benzhydryl ester.

D. 7-Phenoxy-7- (thiazolylacetamide) -cephalosporic acid

A solution of 2 g of 7-phenoxy-7- / 5-thiazolylacetamidodecephalosporanoic acid 7-phenoxy-7- / 5-thiazolylacetamidodephalosporanic acid was stirred at room temperature for 5 min. The excess anisole and trifluoroacetic acid were quickly removed under vacuum. mixed with 50 ml of methylene chloride and extracted twice with sodium bicarbonate solution. The aqueous extract was cooled and covered with a layer of ethyl acetate and the pH adjusted to 2.1 with dilute sulfuric acid. The ethyl acetate layer was dried over sodium sulfate and evaporated to give 7-phenoxy-7- (5-thiazolylacetamide / cephalosporic acid).

Example 67

A. 7-Chloro7- / tert-butoxycarbonylthio / cephalosporic acid benzhydryl ester

Tert-butoxycarbonylsulfenyl chloride is obtained by the addition of tert-butanol to chlorocarbonylsulphenyl chloride in a 1: 1 molar amount at 30 ° C. 10 mmol of tert-butoxycarbonylsulfenyl chloride were dissolved in 50 ml of CH ₂ Cl ₂ and added dropwise to a solution of 10 mmol 7-diazocephalosporanic acid benzhydryl ester in 50 ml of CH ₂ Cl ₂ cooled to -40 ° C in dry ice bathroom. The addition was continued for about 15 min and at the end of the temperature65 the ration began to increase from -10 ° C to -5 ° C gradually. Saturated sodium bicarbonate solution was added and the organic layer was separated and washed with water. After drying with sodium sulfate, the solvent is removed in vacuo. The crude product was analyzed by IR, loss of diazo 2100 cm ^-1 , presence of β-lactam 1790 cm * ¹ , presence of ester band 1745 cm ¹ . It was further purified by TLC to afford the 7-chloro-7-L-butoxycarbonylthio] -cephalosporanic acid benzhydryl ester in pure form.

B. 7-Azido7- (t-butoxycarbonyl and lithio) -cephalosporanic acid benzhydryl ester

To a solution of 5 mmol lithium azide in 5ml dimethylformamide was added 5 mmol 7-chloro7 (t-butoxycarbonylthio) -cephalosporanic acid benzhydryl ester. The solution was heated from 40 ° C to 70 ° C for 3-6 min, then cooled sharply in ice water. The dimethylformamide aqueous solution was extracted with 2 x 25 ml CHCl ₃ , washed with saturated sodium bicarbonate solution, 2 x 50 ml water and the organic layer dried over sodium sulfate. Evaporation of the solvent gave the crude product as a mixture of isomers which can be further purified by chromatography on silica gel. The two isomers were analyzed by IR, the presence of β-lactam 1790 cm ¹ , the presence of an azide band 2100 cm * ¹ , the ester bands 1475 cm ¹ .

C. 7- (2-Thienylacetamido-7 (tert-butoxycarbonylthio) -cephalosporanoic acid benzhydryl ester

The mixture of isomers of the 7-azido compound obtained above (5 mmol) was reduced with 3 g of Bolhoffer catalyst (10% Pd / C) at 40.6,895 kN / m ² at room temperature in 50 ml of ethyl acetate in the presence of 5 mmol pyridine and 5 mmol thienylacetic anhydride.

At the end of the 1st hour, the catalyst was filtered off and the ethyl acetate was extracted with 2 x 20 ml of 1 N HCl and 2 x 50 ml with 10% sodium bicarbonate, 2 x 50 ml of water. The ethyl acetate was dried over sodium sulfate and the solvent was evaporated in vacuo. The crude product, a mixture of isomers, was analyzed by IR, the presence of β-lactam 1790 cm ¹ , the loss of azide 2100 cm * ¹ , the appearance of new amide bonds 1680 cm ¹ .

D. 7-Thienylacetamido-7-mercaptocephalosporanic acid mmol thienylacetamide benzhydryl ester prepared as described was dissolved in 10 ml of anisole and cooled to 0 ° C. To this solution was added 20 ml of trifluoroacetic acid. The solution was kept at 0 ° C and stirred for 1-3 h. Excess trifluoroacetic acid and anisole were removed by evaporation with a vacuum pump. Product evaluated by IR, carboxyl 1710 cm ¹ , β-lactam 1790 cm ¹ , ester strip loss. The compound absorbs in the UV spectrum λ max. ²⁶⁰ * ²⁶⁵ ε / 8000 /.

Example 68

A. 7-Chloro7- (carbamoylthio) -cephalosporanic acid benzhydryl ester

Carbamoylsulfenyl chloride 10 mmol, obtained by the action of 200 mmol of ammonia on chlorocarbonylsulphenyl chloride at -70 ° C in 50 ml of CH ₂ CJ ₂ , was added dropwise to a solution of 10 mmol benzhydryl ester of 7-diazocephalosporic acid. At the end of the addition, the temperature was allowed to rise slowly to -5 ° C. Saturated sodium bicarbonate solution (50 ml) was added and the organic layer was separated, washed with water and dried over sodium sulfate. The solvent was removed in vacuo to give a gum. The crude product was analizi66 pa c IR / absence of diazo 2100 cm ^'1, the presence of a β-lactam a new carbamoyl absorption 1680 ^cm-17. The crude product can be purified by preparative thin layer chromatography.

B. 7-Azido7- (carbamoylthio) -cephalosporanic acid benzhydryl ester

The reaction was carried out in the same manner as for the preparation of 7-azido-7- (tert-butoxycarbonylthio) -cephalosporanic acid benzhydryl ester. The crude product mixture of isomers was analyzed by IR, new azide band 2100 cm ^1, β-lactam 1790 ^cm-1 and a carbamoyl group 1680 cm ^'1.

C. 7-Thienylacetamido-7- (carbamoylthio) cephalosporic acid benzhydryl ester

The reductive acylation is carried out in the same manner as for benzhydryl 7-azido-7- (tert-butoxycarbonylthio) -cephalosporic acid. The crude product was analyzed by IR, loss of azide 2100 cm &lt; -1 ^&gt; , presence of β-lactam, new amide at 1680 cm &lt; ^{-1 &gt;} . It can be further purified by preparative thin layer chromatography or column chromatography.

D. 7-Thienylacetamido-7- (carbamoylthio) cephalosporic acid

The removal of the benzhydryl ester is carried out in the same manner as described for 7-thienylacetamido-7-butoxycarbonylthiocephalosporanic acid. The crude product was analyzed by IR, appears ester band at 1740 cm * ¹ strip acid appears at 1710 ^cm-1 and UV spectrum λ max. ^260-265 ε / 8000 /.

Example 69

A. 7-Bromo-7-methylthiocephalosporanic acid benzhydryl ester mmol 7diasocephalosporanic acid benzhydryl ester was dissolved in 100 ml of methylene chloride and cooled to -40 ° C under nitrogen. To this solution was added 12 mmol of methylsulfenyl bromide in 100 ml of CH ₂ Cl ₂ dropwise with vigorous stirring. Nitrogen is immediately released. After the reagent was added (15 min) at -40 ° C, the mixture was allowed to gradually warm to -5 ° C. Saturated sodium bicarbonate solution was added and the organic layer was separated and washed with water. After drying over sodium sulfate, the solvent is removed at room temperature in vacuo. The crude product was analyzed by IR, loss of diazo 2100 cm * ^1, the presence of β-lactam 1790 ^cm-1 and a positive test for Valshtayn halogen. The crude product can then be purified by preparative TLC or column chromatography.

B. 7-Azido-7-methylthiocephalosporanate mmol benzhydryl ester of 7-bromo-7-methylthio compound is heated at 68 ° C for 4 min in 60 ml of dimethylformamide containing 10 mmol of lithium azide. The solution was diluted with 300 ml of water and extracted with 2 x 50 ml of chloroform. The chloroform layer was washed with 3 x 100 ml water and dried over anhydrous sodium sulfate. The crude product was analyzed by IR, azide 2100 cm &lt; ^{-1 &gt;} , β-lactam 1790 cm &lt; -1 ^&gt; and a negative Balstein test. The crude product was purified by TLC or column chromatography.

C. 7- (2-Thienylacetamido) -7-methylthiocephalosporanoic acid benzhydryl ester mmol of 7-azido-7-methylthio compound is dissolved in 50 ml of ethyl acetate and 10 mmol of thienylacetic anhydride, 0.1 ml of pyridine and 800 g of Bolhofofer are added. . The mixture was hydrogenated at room temperature for 1 h. The catalyst was removed by filtration and the residual residue was evaporated to a glassy mass in vacuo. IR analysis of the crude product showed a loss of azide 2100 cm ¹ , the appearance of a new amide 1680 sit ¹ and βlactam 1790 cm ¹ . The crude product can then be purified by TLC or column chromatography.

D. 7-Thienylacetamide-7-methylthiocephalosporanic acid mmol from 7- (2-thienylacetamido / 7-methylthio compound (mixture of isomers) was dissolved in 10 ml of anisole and cooled to 0 ° C. To the solution was added 15 ml of trifluoroacetic acid cooled to 0 ° C and the mixture was aged at room temperature for 1 h. The excess trifluoroacetic acid and anisole were removed by evaporation in vacuo and the residue was washed twice with chloroform and evaporated to dryness. The crude product was evaluated by IR / ester loss at 1740 cm &lt; -1 ^&gt; and carboxyl at 1710 cm &lt; ¹ & gt ^; / and UV spectrum, λ max. ²⁶⁰ ' ²⁶² ε / 8000 /.

Example 70

7- (2-thienylacetamido) -7-acetylthiocephalosporanic acid. 10 mmol of 7- (2-thienylacetamido-7-mercapto compound was dissolved in 50 ml of pyridine at 0 ° C and acetyl chloride was added dropwise over 5 min. The mixture was cooled sharply in ice water and the pH was adjusted to 8 with sodium hydroxide. The pyridine was removed by ether extraction and the aqueous layer was lyophilized. The crude product was evaluated in the IR / new carbonyl 1740 sated ^1, β-lactam 1790 cm ^'1 and the UV spectrum, λ max. ^2b0 ' ²⁶⁵ is (8000).

Example 71

7- (2-Thienylacetamido-7-methylsulfinylcephalosporanoic acid). 10 mmol of 7- (2-thienylacetamido) -7methylthio compound was dissolved in 50 ml of tetrahydrofuran at 0 ° C and treated with 10 mmol of peracetic acid. The solution was stirred at 0 ° C 30

- - £. · Min. A solution of sodium thiosulfate is added to the K1 paper negative test. 100 ml of saturated sodium chloride solution was added and the organic layer was separated and dried over sodium sulfate. The solvent was removed in vacuo and the product evaluated by IR analysis, β-lactam 1790, sulfinyl bands 1060, 1150 cm ¹ , and UV spectra, λ max. ²⁶⁰ ' ²⁶⁵ .

Example 72

3-Hydroxymethyl7-acetamido-7-methoxydicephalosporanic acid sodium salt

Treatment of 7-acetamido-7-methoxycephalosporanic acid in aqueous solution at pH 6 with acetyl stearase obtained from orange peels yields 3-hydroxymethyl-7-acetamido-7-methoxydicephalosporanic acid, which is isolated as a sodium salt by known procedures .

Example 73

Following the procedure described in Example 72, with the following 7-oxo-7-thiosubstituted cephalosporins, as described in the previous examples, the following 3-hydroxymethyldecephalosporanic acids were obtained.

3-Hydroxymethyl 7-methoxy-7- (2-thiene acetamido) -decephalosporic acid sodium salt; 3-Hydroxymethyl-7-methoxy-7/2-thiophthalene / decephalosporanic acid sodium salt; 3-Hydroxymethyl-7-methoxy-7- (p-guanidinephenylacetamido / decephalosporanoic acid sodium salt; 3-hydroxymethyl-7-methoxy-7- (2-furylacetamido) -decephalosporanoic acid sodium salt; 3-Hydroxymethyl-7-methoxy-7-tetraazolylacetamidodecephalosporanoic acid sodium salt; sodium salt of

3-hydroxymethyl-7-methoxy-7- (O-amino-2-phenylacetamido) -decephalosporic acid; sodium salt of

3-hydroxymethyl-7-methoxy- (2-carboxy-2-phenylacetamido) -decephalosporic acid; sodium salt of

3-hydroxy methyl 1-7-benzyloxy-phenylacetamidodecephalosporanoic acid; 3-Hydroxymethyl-7-ethoxy-7- (2-thienylacetamido / decephalosporanoic acid sodium salt; 3-Hydroxymethyl-7-methylthio-7-phenylacetamidodecephalosporanoic acid sodium salt; 3-Hydroxymethyl-7-mercapto-7- (2-thienylacetamido) -decephalosporanoic acid sodium salt; 3-Hydroxymethyl7-carbamoylthio-7- (2-thienylacetamido) -decephalosporanoic acid sodium salt; 3-Hydroxymethyl-7- (2-thienylacetamido-7-aminocarbonyloxy) -decephalosporanoic acid sodium salt; 3-hydroxymethyl-7- (2-thienylacetamido) -7- (methoxycarbonyloxy) -decephalosporanoic acid sodium salt; 3-Hydroxymethyl 7- (2-thienylacetamido) -7-aminosulfonyloxy] -decephalosporanoic acid sodium salt.

Example 74

3- (N, N-Dimethylcarbamoyloxymethyl) -7-methoxy-7- (2-thienylacetamido) -decephalosporanoic acid sodium salt

Excess phosgene was bubbled into a stirred solution of 0.5 g of 3-hydroxymethyl-7-methoxy-7- (2-thienylacetamido) -cephalosporanic acid in 100 ml of methylene chloride and the resulting mixture was allowed to stand at room temperature overnight. The excess phosgene was removed by bubbling dry nitrogen through the solution for 3 h and the resulting solution was evaporated under reduced pressure.

To a solution of 0.45 g of the residue in 50 ml of cooled methylene chloride was added 0.2 g of dimethylamine. The mixture was stirred for one hour at room temperature and excess amine hydrochloride was removed by filtration. To the resulting filtrate was added a solution of 0.1 g of sodium bicarbonate in 20 ml of water. The mixture was stirred for one hour at room temperature and the aqueous layer was separated, washed twice with methylene chloride and dried in cold. A mixture containing 3-N, N-dimethylcarbamoyloxymethyl-7-methoxy-7- (2-thienylacetamido) -decephalosporanic acid sodium salt was obtained.

Similarly, the other 3-hydroxymethyldecephalosporanic acids obtained in Example 73 were converted to the corresponding salts of 3- (N, N-dimethylcarbamoyloxymethyl) -decephalosporanic acid.

Example 75

Following the procedure described in Example 74, and using an equivalent amount of piperidine, pyrrolidine or morpholine instead of dimethylamine, the following products are obtained: 3- (piperidinecarbonyloxymethyl) -7-methoxy-7- (2-thienylacetamido) -decephalosporic acid sodium salt ; 3-Pyrrolidinylcarbonyloxymethyl] -7-methoxy-7- (2-thienylacetamido-decephalosporanoic acid sodium salt; 3-morpholinecarbonyloxymethyl-7-methoxy-7- (2-thienylacetamido) -decephalosporanoic acid sodium salt; 3- (piperidinecarbonyloxymethyl) -7-methoxy-7methylthio-7- (2-thienylacetamido) -decephalosporanoic acid sodium salt.

Example 76

3- (N-Methylcarbamoyloxymethyl) -7-methoxy-7 (2-thienylacetamido) -decephalosporanoic acid sodium salt

The 3-hydroxymethyl-7-methoxy-7- (2-thienylacetamido / decephalosporanoic acid) sodium salt of 250 mg was suspended in 5 ml of dimethylformamide. To this mixture was added, with stirring, 0.2 ml of trimethylamine and 0.7 ml of methylisocyanate, and the resulting reaction mixture was allowed to stand for 1/2 hour, then evaporated to dryness under reduced pressure. The resulting residue was mixed with 10 ml of ethyl acetate and 10 ml of 0.1 N phosphate buffer. The pH of the aqueous layer was adjusted to 1.6 and the mixture was stirred for 2 h at room temperature. The pH was then adjusted to 8 with aqueous tricalcium phosphate and the aqueous phase separated. The organic phase was re-extracted with 10 ml of phosphate buffer at pH 8. The combined aqueous phases were adjusted to pH 2.1 with hydrochloric acid and extracted twice with ethyl acetate. The ethyl acetate extracts were treated with 10 ml of water containing 0.15 g of sodium bicarbonate. . Separation and drying in the cold of the aqueous phase afforded the 3- (N-methylcarbamoyloxymethyl) -7-methoxy-7- (2-thienylacetamido) -decephalo5-sporic acid sodium salt.

Similarly, the 3-hydroxymethylcephalosporanates obtained in Example 73 are converted to the corresponding

3- (L-methylcarbamoyloxymethyl) compound.

Example 77-86

According to the procedure of Example 76 and using other isocyanates, the new 3-hydroxymethyl-7-oxysubstituted cephalosporan compounds were converted to the corresponding N-substituted 3-carbamoyloxymethyl compounds according to the following scheme

CH ₂ OH + OC

-NR.,

->

ο II

CH ₂ OCNHR ₁₇

COONa

COONa

Example axis = nr _I7 «Π 77 -axis - nch ₂ ch ₂ ci -CH ₂ CH ₂ CI 78 -axis = nch ₂ ci -CH ₂ CI 79 OC = NC / CH _3/3 -s / dream ₃ /, 80 OS = nc ₂ h ₂ -with ₂ n ₅ 81 axis = nc / ch _3/2 ch ₂ ci -c / ch _3/2 chci 82 axis = ncooc ₂ h _s -coax ₂ n ₅ 83 OC = NSO - ^CH 3 84 OS = -ο 85 OS = ΝΟΗ- / θ / ₂ ' ^{CH_ /} O ^{/ 2} 86 OS = N- ^ ~~ ^ -SO- -Ο-so, -

in which R 'and R ₍ have the meanings shown in Example 73.

Example 87

3-Pyridinomethyl-7-methoxy-7- (270 furylacetamido) -decephalosporic acid

A solution of 1 g of sodium methoxy-7- (2-furylacetamido / cephalosporanic acid sodium salt in 5 ml of water was brought to pH 2.5 with dilute HCl. 8 ml of pyridine was added and the solution heated at 70 ° C for three and a half hours. The resulting reaction mixture was lyophilized and the residue dissolved in water and passed through a polystyrene trimethylammonium anion exchange resin (43% water). The resulting adsorbate on the resin was eluted with water and the selected fractions lyophilized to give pure 3-pyridinomethyl-7-methoxy-7- (2-furylacetamido / decephalosporic acid.

When an equivalent amount of trimethylamine or triethylamine is substituted with pyridine according to the above procedure, otherwise the procedure described is followed, the corresponding 3- (trimethylammoniummethyl) and 3- (triethylammoniummethyl) compounds are obtained.

Similarly, when 3-fluoropyridine, 4-trifluoromethylpyridine, 3carboxypyridine, 4-carbamoylpyridine, 4- (N-methylcarbamoyl) pyridine,

4- / N, N-dimethylcarbamoyl / pyridine,

3-Carboxymethyl / pyridine, 2-methylpyridine, 3- (hydroxymethyl) pyridine, 3-sulfopyridine, or 3-cyanpyridine are substituted with pyridine according to the procedure described, to give the corresponding 3 or 4-substituted pyridine methyl compounds.

Example 88

3-Thiouronomethyl-7-methoxy-7-phenylacetamidodecephalosporic acid

A solution of 1 g of 7-methoxy-7-phenylacetamidocephalosporanic acid and 1 g of thiourea in 25 ml of water was maintained at 37 ° C for 5 days. Acetone (200 ml) was added and the mixture was cooled in an ice bath. The cooled solution was filtered to capture the precipitated product, which was then fractionated through polystyrene trimethylbenzylammonium anion exchange resin with 43% water. Selected fractions were lyophilized and the crude product was then crystallized from methanol / water to give

3-Thiouronomethyl-7-methoxy-7-phenylacetamidodecephalosporanoic acid. Substitution of the equivalent amount of N-methylthiourea, N-ethylthiourea, N, N-dimethylthiourea or N, N-dipropylthiourea in the previous process instead of thiourea, and following the procedure described, the corresponding N-substituted compounds are obtained.

Example 89

3- (ethylthiomethyl) -7-methoxy-7- (2-thienylacetamido) decefalosporic acid

A mixture of 7-methoxy-7- (2-thienylacetamido / cephalosporic acid 0.65 g and ethanethiol 0.37 ml in 10 ml of 50% aqueous acetone was stirred at room temperature and a 10% aqueous solution of 2 ml of sodium hydroxide was added at stirring. The resulting reaction mixture was then heated in a sealed glass tube for 100 h and the resulting mixture was concentrated under reduced pressure. The residue was dissolved in water and fractionated through polystyrene trimethylammonium anion exchange resin, 43%. Selected fractions were combined and lyophilized to give 3- (ethylthiomethyl) -7-methoxy-7- (2-thienylacetate) and to (N-decephosphoric acid).

After substitution of an equivalent amount of methanethiol, propanethiol, pyridine-2-thiol, pyridine-3-thiol, pyridine-4-thiol, benzothiazole-2-thiol, 4-methylpyrimidin-2-thiol or 271 methyl-3,4-thiadiazole -5-thiol instead of ethanethiol according to the above procedure, according to the procedure described above are obtained respectively

3- (methylthiomethyl), 3- (propylthiomethyl), 3- (2-pyridylthiomethyl), 3- (3-pyridylthiomethyl), 3- (4-pyridylthiomethyl), 3- (2-benzthiazolylthiomethyl),

3- (4-methylpyrimidin-2-ylthiomethyl) and 3- (2-methyl-3,4-thiadiazol-5-ylthiomethyl) substituted 7-methoxy-7/2-thienylacetamido / decephalosporic acid.

Example 90

3-N, N-dimethylthiocarbamoylthiomethyl] -7-methoxy-7-tetrazolylacetamide decalphosphoric acid

A. A solution of 6.8 g of 7-methoxy-7-tetrazolylacetamidcephalosporanic acid and sodium N, N-dimethyldithiocarbamate 2.8 g in 60 ml of water is heated to 50 ° C for 24 hours. The product is lyophilized and then fractionated through polystyrene trimethylbenzylammonium anion exchange resin (43% water). Selected fractions were then lyophilized to give 3-N, N-dimethylthiocarbamoylthiomethyl) -7-methoxy-7-tetrazolylacetamidodecephalosporic acid.

After substitution of an equivalent amount of the following reagents: N-methyldithiocarbamic acid sodium salt, N, N-diethyldithiocarbamic acid sodium salt, N, N-di-n-propyldithiocarbamic acid sodium salt, N-methyl-1H-sodium salt (2-dimethylaminoethyl) -dithiocarbamic acid, M-ethyl-n- (2-diethylaminoethyl) -dithiocarbamic acid sodium salt, N- (2-di-n-propylaminoethyl-dithiocarbamic acid sodium salt, M-methyl- > 4- (2-morpholinethyl) ethiocarbamic acid, sodium salt of N-methyl-N / 3-diethyl nopropyl / dithiocarbamic acid, Nphenyl-N- (2-methylaminoethyl) dithiocarbamic acid sodium salt, N, N-tetramethyldithiothiocarbamic acid sodium salt, N, pentamethylenedithiothiocarbamic acid, N, N -hydroxy-6-sodium, N- dithiocarbamic acid or the sodium salt of 4-methylpiperazine dithiocarboxylic acid with the sodium salt of dimethyldithiocarbamic acid according to the above procedure, following the procedure described, the corresponding 3- (N, N-diethylthiocarbamoylthiomethyl), 3- (N-N, N-N, N-diethylthiocarbamoylthiomethyl) are obtained. prop thiocarbamoylthiomethyl (-3- (N-methyl-N- (2-dimethylaminoethyl) thiocarbamoylthiomethyl, 3- (N-ethyl-L4) 2-di-n-propylaminoethyl) thiocarbamoylthiomethyl), 3 (N-methyl-1H- (2-morpholinoethyl)) thiocarbamoylthiomethyl), 3- (N-methyl-N (3-diethylaminopropyl) thiocarbamoylthiomethyl), 3- (N-phenyl-N- (2methylaminoethyl) thiocarbamoylthiomethyl), 3- (N, N-tetramethylene (thiocarbamoylthiomethyl), 3- (N-tetramethyl) thiocarbamoylthiomethyl) , N-pentamethylene / thiocarbamoylthiomethyl, 3- (N, Nbis- (2-hydroxyethyl) thiocarbamoylthiomethyl) and 3- (4-methylpiperazine) thiocarbamoylthiomethyl, 7-methoxy-7tetrazolylacetamidodecephalosporic acid.

Similarly, other 7-substituted cephalosporins, prepared as described in the previous examples, were converted to the corresponding 3-substituted compounds as described above.

Example 91

3- (benzoylthiomethyl) -7-methoxy-7/2-carboxy-3-phenylacetamido / decephalosporic acid

A mixture of 0.654 g of 7-methoxy-7- (2-carboxy-2-phenylacetamido / cephalosporic acid, 0.504 g of sodium bicarbonate and 0.414 g of thiobenzoic acid in 5.0 ml of water was heated at 50 ° C overnight under a nitrogen atmosphere. . The product was precipitated by the addition of acetone and crystallized from a mixture of alcohol and water to give 3- (benzoylthiomethyl) -7methoxy-7- (2-carboxy-3-phenylacetamido) decephalosporic acid.

After substitution of an equivalent amount of sodium salt of ethylxanthogenic acid, potassium salt of n-propylxanthogenic acid, potassium salt of n-hexylxanthogenic acid, potassium salt of isopropylxanthic acid, n-butyl acid cyclopentanoic acid cyclohexylxanthogenic acid salt with thiobenzoic acid according to the above process, and following the procedure described above, the corresponding 3 (ethoxythiocarbonylthiomethyl), 3- (n-propoxythiocarb) is obtained niltiometil / 3izopropoksitiokarboniltiometil, 3 / izopropoksitiokarboniltiometil, 3 / -H-butoksitiokarboniltiometil / 3 / n-heksiloksitiokarboniltiometil /

3- (- cyclopentyloxythiocarbonylthiomethyl) and 3- (cyclohexylthiocarbonylthiomethyl) -7-methoxy-7- (2-carboxy-

2-phenylacetamido / decephalosporanic acids.

Similarly, the other 7-substituted cephalosporins prepared as described in the above examples can be converted to the corresponding 3-substituted compounds.

Example 92

3- (toluene-p-sulfonylmethyl) -7-methoxy-7- (2-thienylacetamido) decephalosporic acid

A mixture of 0.654 g of 7-methoxy-7- (2-thienylacetamido / cephalosporanic acid and 1.0 g of sodium salt of tolu-en-sulfinic acid in 5 ml of water was heated at 50 ° C for 24 hours. The mixture was concentrated in vacuo. and crystallized from a mixture of methanol and water to give 3- (toluene-p-sulfonylmethyl) -7-methoxy-7- (2-thienylacetamido) decephalosporic acid.

Example 93

3- (azidmethyl) -7-methoxy-7- (2-furylacetamido) -decephalosporic acid

A mixture of 2.0 g of 7-methoxy-7- (2-furylacetamido / cephalosporanic acid and 1.0 g of sodium azide was dissolved in 10 ml of water and heated at 50 ° C overnight. The mixture was then lyophilized to give crude 3-azidomethyl) -7-methoxy-7- (2-furylacetamido) -decephalosporic acid.

Alternatively, instead of treating 7-methoxy-7- (2-furylacetamido) cephalosporanic acid with sodium azide, it is possible to substitute 3- (carbamoyloxymethyl) -7- (2-furylacetamido) -7-methoxydicephalosporanic acid by an otherwise analogous method, such as an identical product is obtained. The following examples illustrate the process for the preparation of 3 (carbamoyloxymethyl) -7- (2-furylacetamido) -7-methoxydicephalosporanic acid (100 mg) in 0.5 M phosphate buffer solution / 5 ml, obtained by the addition of 3.5 g sodium dihydrogen phosphate and 3.4 g of disodium phosphate in 100 ml of water, followed by sufficient hydrochloric acid to bring the pH to 5 / heated in the presence of sodium azide (20 mg) at 95 ° C for 8 min. After preparative TLC. afforded 20 mg of 3 (azidmethyl) -7-methoxy-7- (2-furylacetamido) decefalosporanic acid as sees from IR and MRI the identification. Treatment of this material with 1.0 ml of trifluoro73 acetic acid at 0 ° C for 5 min followed by termination of the high volume ether reaction and evaporation of the solvent gave 15 mg of 3- (azidomethyl) -7-methoxy-7 - / 2furylacetamido / decephalosporanic acid.

Example 94

3- (2,4-dihydroxybenzyl) -7-methylthio-7-phenylacetamidodecephalosporic acid

A mixture of 0.654 g of 7-methylthio-7-phenylacetamidocephalosporanic acid, 1.1 g of resorcinol and 10 ml of water was heated at 50 ° C for 2 days. The reaction mixture was then lyophilized to give

3- (2-, 4-Dihydroxybenzyl) -7-methylthio-7-phenylcetone midodecephalosporic acid.

Example 95

3- [1-methylindol-3-yl- [7-benzyloxy-7-phenylacetamidocephalosporic acid]

A solution of 0.655 g of N-methylindole in 5 ml of acetone was added to a solution of 0.654 g of 7-benzyloxy-7-phenylacetamidocephalosporanic acid in water 5 ml at 50 ° C. The mixture was heated for 48 h and then the solvent was removed in vacuo and the residue triturated with ether to give 3- (N-methylindol-3-yl) 7-benzyloxy-7-phenylacetamidodecephalosporanoic acid.

Example 96

3-Methyl-7-methoxy-7/2-thienylacetamido / decephalosporic acid

The 10% palladium-on-carbon catalyst was suspended in water, 80 ml and treated with hydrogen. The catalyst was then filtered and resuspended in water 50 ml and to this mixture 2.67 g was added 7-methoxy-7/2-thienylacetamide / cephalosporic acid sodium salt (1.0g) in water 10 ml. The resulting mixture was shaken for 22 h at room temperature.

The catalyst was removed by filtration and washed once with 50 ml water. The combined washing liquid and filtrate were then concentrated in vacuo to dryness to give 3-methyl-7-methoxy-7- (2-thienylacetamido / decephalosporic acid.

Example 97

3- (amidine thiomethyl) -7-methoxy-7/2-thienylacetamido / decephalosporic acid

100 mg of 7-methoxy-7- (2-thienylacetamido / cephalosporanic acid was heated for 8 min with thiourea, 26 mg at 95 ° C in 0.5 M phosphate buffer solution / 5 ml, obtained by the addition of 3.5 g sodium dihydrogen phosphate and 3.4 g of disodium phosphate in 100 ml of water, followed by the addition of sufficient hydrochloric acid to bring it to pH 5 /. Electrophoresis of the solution at pH 7 shows the thiuron compound as a fixed whole. The mixture was purified by absorption on a polystyrene nuclear sulfonic acid cation exchange resin in a hydrogen cycle (Dowex 50) to remove the phosphate buffer. The elution was performed using a 0.1 N pyridine solution. The pH was adjusted to 8 with 1N sodium hydroxide and evaporated in vacuo to remove residual pyridine. Freeze-drying gave the title compound.

Example 98

3- (4-methylthiazol-2-ylmercaptomethyl) -7-methylthio-7- (2-thienylacetamido) decephalosporic acid

7-methylthio-7- (2-thienylacetamido / cephalosporic acid, 100 mg in 5 ml buffer with pH 7 / 0.5 M solution of a mixture of 3.5 g of sodium dihydrogen phosphate and 3.4 g of disodium phosphate in 100 ml of water / containing 2-mercapto74

4-methylthiazole, 50 mg was heated at 95 ° C for 8 min. The resulting reaction mixture contained the title compound.

Example 99

3- (1,3,4-Thiadiazol-2-ylmercaptomethyl) -7-methoxy-7- (2-thienylacetamido) decephalosporic acid

By reacting 2-mercapto-1,3,4-thiadiazole instead of 2-mercapto-4-methylthiazole and 7-methoxy-7- (2-thienylacetamido / cephalosporic acid and following the procedure described in Example 98, the title compound was obtained .

Example 100

3- (thiocyanatomethyl) -7-methoxy-7/2-furylacetamido / decephalosporic acid

7-Methoxy-7- (2-furylacetamido / cephalosporanic acid, 100 mg was added to a 0.5 M buffer solution of 5 ml consisting of 3.5 g of sodium dihydrogen phosphate and 3.4 g of disodium phosphate in 100 ml of water and pH The mixture was adjusted to pH 5 by the addition of hydrochloric acid. Sodium thiocyanate, 20 mg was added and the mixture was heated at 95 ° C for 8 min. This gave the title compound.

Example 101

3- (chloromethyl) -7-methoxy-7- (2-thienylacetamido) decefalosporanic acid in-trifluoroacetate

7-Methoxy-7- (2-thienylacetamido / cephalosporanic acid benzhydryl ester, 1mmol was dissolved in methylene chloride, 5 ml and the mixture was cooled to 0 ° C. Colidine 1 mmol was added and then a solution of phosphorous pentachloride 0.6 mmol in methylene chloride 5 ml was added dropwise. The mixture was then stirred for one hour in an ice bath at 0 ° C and the resulting solution extracted with sodium bicarbonate, dilute hydrochloric acid and saturated sodium chloride solution. The mixture was evaporated to dryness and then isolated by chromatography on a cooled silica gel column using chloroform as eluent. The product obtained is 3- (chloromethyl) -7- (2-thienylacetamido) decefalosporanic acid benzhydrol ester.

A solution of this benzhydryl ester (1 mmol) in anisole, 13 ml, was poured into 6.5 ml of cold (0 ° C) trifluoroacetic acid with stirring. After 5 min, the solution was poured under stirring into an ethereal solution, 1800 ml, maintained at 0 ° C. The solid precipitate which is obtained is collected and dried to give the title compound.

Example 102

3- (N- (2-Chloroethyl) carbamoyloxymethyl) -7-methoxy-7- (2-thienylacetamido) decalphosphoric acid sodium salt

A suspension of 100 mg of 3-hydroxymethyl-7- (2-thienylacetamido / decephalosporic acid as potassium salt was placed in 3 ml of dimethylformamide under nitrogen and stirred by ultrasonic waves. 0.1 M triethylamine and 0.16 ml β-chloroethylisocyanate were added. After 2 h, the solution was diluted with ethyl ether and centrifuged. The ether was decanted and the oily residue was washed with more ether. After centrifugation again, the ether was decanted again. The solid residue was dissolved in water and the pH was adjusted to 2 with concentrated HCl. The product was extracted with ethyl acetate, which was pre-washed with 5% sodium bicarbonate solution. The ethyl acetate was dried with magnesium sulfate, filtered and evaporated. The residue was redissolved in ethyl acetate and washed with an aqueous solution of 16 mg of sodium bicarbonate, the pH of the solution was adjusted to 7.6 with sodium bicarbonate and the mixture was stirred for half an hour. The layers were separated and the aqueous solution was washed with ethyl acetate and then lyophilized overnight.

g of the crude product obtained is dissolved in methanol and all the insoluble material is filtered off. The methanol was evaporated and a little ethyl ether was added to begin the initial solidification. Yield 55 mg, one spot on TLC, K _f 0.54 in butanol: ethanol: water / 4: 1: 5 / top layer.

Example 103

3- (N- (4-sulfenyl) carbamoyloxymethyl) -7-methoxy7- (2-thienylacetamido) decefosporanoic acid disodium salt

A suspension of 200 mg of the 3- (hydroxymethyl) -7-methoxy- (2-thienylacetamido) -decephalosporanoic acid potassium salt in 5 ml of dimethylformamide was placed under nitrogen and stirred with ultrasonic waves. 0.096 mg of triethylamine and then 94.4 mg of p-chlorosulfonylphenylisocyanate in 1 ml of dimethylformamide are added. After 5 min, the solution was diluted with ethyl ether and centrifuged. The ether was decanted and the green solid was washed with more ether. After centrifugation again, the ether was decanted again. The resulting solid was dissolved in 30 ml of water containing 50 mg of NaHCO _r. After stirring for 1/2 hour, the pH was adjusted to 7.1 with dilute HCl. The solution was freeze-dried overnight to give 295 mg of crude product which was dissolved in hot methanol and filtered from insoluble metal. A small amount of isopropanol is added and the first precipitate is filtered off and discarded. More isopropanol is then added and the product is obtained twice with 39 mg and 74 mg of the title compound. Electrophoresis in 10% acetic acid shows one spot.

Example 104

A. 7β-6ροмо-7-α-methoxycephalosporanic acid benzhydryl ester

To a solution of 1.8 g of 7-diazocephalosporanic acid benzhydryl ester in 200 ml of methylene chloride was added a cold solution of 560 mg of N bromoacetamide in 20 ml of methanol. The mixture was stirred at room temperature for 30 min and the solvent was quickly removed under vacuum with a water pump. The resinous residue was stirred with methylene chloride and washed with sodium bicarbonate solution, the organic phase was dried over anhydrous magnesium sulfate and evaporated. The oily residue was chromatographed on 60 g of silica gel. Elution with methylene chloride affords 7β-6ροΜ-7-α-ΜετοκοΗcephalosporanic acid benzhydryl ester.

B. 7-a-Azido-7? -Methoxycephalosporanic acid benzhydryl ester

7f-Bromo-7-a-methoxycephalosporanic acid benzhydryl ester solution, 700 mg and lithium azide, 435 mg in 5 ml of dimethylformamide was stirred at 30-35 ° C for 6 hours. The dimethylformamide was evaporated under high vacuum and the residue was mixed with methylene chloride and extracted four times with water. The methylene chloride phase was dried over anhydrous sodium sulfate and evaporated. The oily residue was chromatographed on 20 g of silica gel. Elution with hexamethylene chloride yields 7-a-azido7-β-methoxycephalosporanic acid benzhydryl ester.

C. 7-Amino-7 ^ -methoxycephalosporanoic acid benzhydryl ester

7-a-azido benzhydryl ester76

7-fluoro-methoxycephalosporanic acid, 2 g and platinum oxide, 2 g in 200 ml of dioxane are vigorously stirred under a hydrogen atmosphere for 2 hours. The dioxane is evaporated under reduced pressure, chloroform is added to the residue and filtered through 20 g of silica gel, in a porous-bottomed funnel. The filter cake was washed with 500 ml of chloroform and the combined filtrate and washes were evaporated to give 7-amino-β-methoxycephalosporanic acid benzhydryl ester.

D. 7-β-β-Ethoxy-7-α- (2-thienylacetamido) -cephalosporanoic acid benzhydryl ester

To an ice-cooled solution of 0.9 g of 7-a-amino-7-β-methoxy cephalosporanic acid benzhydryl ester in 20 ml of methylene chloride was added 0.5 ml of dry pyridine and then 0.15 thienylacetyl chloride in 5 ml of cold methylene chloride. The mixture was stirred in an ice bath for 10 min, then washed sequentially with pH 2 phosphoric acid buffer, water and sodium bicarbonate solution. The methylene chloride phase was dried over anhydrous sodium sulfate and evaporated and the residue was chromatographed on 20 g of silica gel. Elution with chloroform afforded 7H-methoxy-7a- (2-thienylacetamido) -cephalosporanic acid benzhydryl ester.

E. 7-β-Μετοκοπ-7α- (2-thienylacetamido) -cephalosporanic acid sodium salt. A solution of 0.8 g of 7H-methoxy-7a- (2-thienylacetamido) -cephalosporanoic acid benzhydryl ester in 3 ml of anisole and 7 ml of trifluoroacetic acid was stirred at room temperature for 5 min and the excess anisole and trifluoroacetic acid removed quickly. by suction of a vacuum pump. A mixture of methylene chloride and water was added to the residue and stirred rapidly until sodium bicarbonate was added to pH 6.5. The aqueous layer was separated, washed with ether and lyophilized to give the 7'-methoxy-7-a- (2-thienylacetamide) -cephalosporanic acid sodium salt.

F. Epimerization of 7H-bromo-7-a-methoxycephalosporanic acid benzhydryl ester. A solution of 250 mg of 7-βbromo-7-a-methoxycephalosporanic acid benzhydryl ester and 0.2 g of lithium bromide in 1.5 ml of dimethylformamide was stirred at room temperature overnight. The dimethylformamide was removed in a vacuum pump and the residue was taken up in chloroform and washed with water. The chloroform was evaporated to give an equilibrium mixture containing predominantly the 7-abrom-7-methoxycephalosporanic acid benzhydryl ester. It is separated from the starting epimer by chromatography on silica gel.

G. 7'-Azido-7a-methoxycephalosporanoic acid benzhydryl ester

This product was prepared as indicated in Example B, starting from α-bromo-7-β-methoxycephalosporanic acid benzhydryl ester. The product is identical in every respect to the 7β-azido-7-α-methoxyacetic acid benzhydryl ester previously prepared by the action of silver fluoroborate in methanol on the 7-azido-7-bromocephalin benzhydryl ester.

Example 105

3carbamoyloxy-7-a-methoxy-7- (2-thienylacetamido) -decephalosporanoic acid pivaloyloxymethyl ester.

A mixture of 0.90 g of the sodium salt of 3carbamoyloxy-7-a-methoxy-7- (2-thi77 enylacetamido) -decephalosporic acid, 0.3 g of chloromethylpivalate and 5 ml of dry dimethylformamide was shaken overnight at room temperature. The reaction mixture was then poured into 25 ml of phosphate buffer with pH 6 and extracted with 3 x 10 ml of chloroform. The combined extracts were washed with 4 x 10 ml water and 10 ml saturated sodium chloride solution and dried over anhydrous magnesium sulfate. Evaporation of the solvent in vacuo afforded a gum which was washed by decantation with petroleum ether to give the 3-carbamoyloxy7-a-methoxy-7- (2-thienylacetamide / cephalosporic acid pivaloyloxymethyl ester.

Example 106

A. 7- (Rnitrobenzylidenamino) -7-trifluoromethoxycephalosporanate benzhydryl ester

7- (p-Nitrobenzylidenamine) -cephalosporanic acid benzhydryl, 571 mg was stirred at 0 ° C under nitrogen in 10 ml of acetonitrile. Vis (Trifluoromethyl) -peroxide, 170 mg was added and then 378 mg of diisopropylethylamine in 5 ml of acetonitrile was added over 1 hour.

The reaction mixture was evaporated in vacuo, mixed with 25 ml of benzene, filtered and washed sequentially with water, dilute phosphoric acid, buffered at pH 2, water and aqueous bicarbonate. The solution was dried with magnesium sulfate, filtered and evaporated to give 7 - [- p-nitrobenzylidenamino] -7-trifluoromethoxy cephalosporic acid benzhydryl ester as a mixture of epimers at C-7. The product was separated by chromatography on silica gel, eluting with 4: 1 chloroform-ethyl acetate.

B. 7-3 ~ Amino-7-a-trifluoromethoxycephalosporas benzhydryl ester new acid, hydrochloride. The 7β- (ρnitrobenzylidenamine) -73-trifluoromethoxy isomer 655 mg and 260 mg aniline hydrochloride were stirred together for 5 h at 250 ° C in 10 ml of methanol. The methanol was removed at 0.1 mm pressure and 30 ° C and the residue was coated with ether to crystallize for 1 h. The solid was treated with ether, filtered and washed with ether several times. The product consists of the title compound and aniline hydrochloride, mixed together and used immediately for next time.

The other epimer is converted to the epimeric compound in the same way.

C. 7β- (2-thienylacetamide) -7-a-trifluoromethoxycephalosporanic acid benzhydryl ester. The mixture of product and aniline hydrochloride obtained as described in B was stirred vigorously at -10 ° C in 25 ml of methylene chloride. Thienylacetyl chloride, 0.5 g was added and then 0.5 g of triethylamine was slowly added. This results in the free amine, the benzhydryl ester of 7β-βθ-7-αtrifluoromethoxycephalosporanic acid, which is immediately acylated. The reaction mixture was allowed to warm slowly to room temperature. The excess acid chloride is hydrolyzed by shaking with water and then the methylene chloride layer is washed successively with dilute phosphoric acid, buffered to pH 2, water and dilute bicarbonate. After drying with magnesium sulfate, the solution was filtered off and evaporated to give a mixture of the product and N-phenyl thienylacetamide. Pure product was obtained after silica gel chromatography, eluting with 4: 1 chloroformethyl acetate.

The other epimer at C-7 is converted to the epimeric compound using the same procedure.

D. 73- (2-Thienylacetamide) -7a-trifluoromethoxycephalosporanic acid sodium salt. 73- (2-Thienylacetamide) -7-atrifluoromethoxycephalosporanic acid benzhydryl ester, 646 mg was dissolved in 0.8 ml of anisole and cooled to 0 ° C. Trifluoroacetic acid 4 ml was added, cooled to 0 ° C, and the reaction was allowed to react at 0 ° C for 2 min. A pressure of 0.1 mm is immediately applied and the reaction mixture is allowed to warm to room temperature. The anisole was then distilled at 30 ° C / 0.1 mm. Add a few ml of anisole to the residue and withdraw at 30 ° C / 0.1 mm to ensure removal of all trifluoroacetic acid. The product was treated with 5 ml of water containing 84 mg of NaHCO ₃ and lyophilized. The resulting powder was washed thoroughly with ether and dried to give the title compound.

The sodium salt of the other epimer at C-7 is obtained in the same way.

Example 107

7- (O-α-sulfoaminophenylacetamide) -7-methoxycephalosporanic acid disodium salt

A suspension of 10 mmol 7- (O-α-aminophenylacetamide) -7-methoxycephalosporanic acid in 100 ml of methylene chloride containing 12 mmol triethylamine was stirred and cooled in an ice bath. 12 mmol of triethylaminosulfur trioxide were added in small portions over 5 minutes. After 1/2 hour the mixture was filtered and concentrated to 10% of the original volume. The solution was diluted with an equal volume of acetone and 30 mmol of 2-ethylhexanoic acid sodium salt was added. Solid product is obtained by sharp cooling and scratching and is collected by filtration. The 7-N-α-sulfoaminophenylacetamide-7-methoxycephalosporanic acid disodium salt can be recrystallized from isopropanol-water if desired.

Example 108

7a-cyano-7-2carboxyphenylacetamide / cephalosporic acid disodium salt.

A. 7a-cyan-7-azidocephalosporanoic acid benzhydryl ester

To a solution of benzhydryl 7-bromo-7-azidotsefalosporanova acid 0,543 g, 0,001 mmol in 10 ml CH ₃ CN was added a solution of 0,350 g of tetrabutylammonium cyanide in 15 ml CH ₃ CN. The reaction mixture was stirred at room temperature overnight, diluted with CH ₂ Cl ₂ and washed with water, dried and evaporated. Chromatography on silica gel gave 7α-cyan-7-azidocephalosporanic acid benzhydryl ester and 7β-πΗ3Η-7-33ΗΛθπεphalosporanic acid benzhydryl ester.

Tetrabutylammonium cyanide is prepared as follows:

g of tetrabutylammonium iodide was dissolved in 10 ml of 20% aqueous NaCN. The aqueous phase is discarded. The organic phase is treated with 3 and then 5 ml of NaCN solution. The organic phase was dried over magnesium sulfate and evaporated to give tetrabutylammonium cyanide.

B. 7a-cyan-7-aminocephalosporanoic acid benzhydryl ester

0.500 g of benzhydryl 7a-cyan-7-azidocephalosporanic acid were dissolved in 50 ml of ethyl acetate, 0.500 g on a 10% palladium on carbon catalyst was added and the mixture was stirred under hydrogen overnight. The catalyst was filtered off and evaporated and the residue was chromatographed on silica gel to separate the starting material from the final product.

C. 7a-cyan-7- (2-benzhydryloxycarbonyl-phenylacetamide) -cephalosporic acid benzhydryl ester. 0.473 g of 7a-cyan-7-aminocephalosporanic acid benzhydryl ester was dissolved in 25 ml of CH ₂ Cl ₂ and cooled to 0 ° C. 2-Benzhydryloxycarbonylphenylacetyl chloride 0,400 g was then added and 0,200 g of pyridine was added after 1 min. The reaction mixture was stirred at 0 ° C for 25 min and poured onto crushed ice. The mixture was stirred and the organic phase separated and washed once with 5% NaHCO ₃ solution, once with pH 2 phosphate buffer and once with water. The organic phase was dried over sodium sulfate and evaporated to give a crude product. The purified product was obtained after chromatography on silica gel.

D. 7a-cyano-7/2-carboxyphenylacetamide / -cephalosporanic acid disodium salt. 7a-cyan-7- (2-benzhydryloxycarbonylphenylacetamide / cephalosporic acid benzhydryl ester) 0.350 g was dissolved in 3 ml of anisole and treated with 10 ml of trifluoroacetic acid at room temperature for 10 minutes. ° C and the residue was mixed with 25 ml of methyl isobutyl ketone and treated with 0.300 g of NaHCO ₃ in 30 ml of water. The mixture was stirred for 1/2 hour, the organic phase was separated, and the aqueous phase was washed twice with methylene chloride and then lyophilized. The solid was crystallized from methanol / isopropanol.

Example 109

A. 7-Azido7-carboxycephalosporanic acid benzhydryl ester

7-Azido-7-bromocephalosporanic acid benzhydryl ester, 5.43g, 0.01 mmol was dissolved in 20 ml of dry ether and cooled to -20 ° C and slowly, with vigorous stirring, 10 ml of 1M phenyllithium solution was added. After 1 h at room temperature, the mixture was poured onto spray dry ice. The residue was extracted with water and acidified to give 7-azido-7-carboxycephalosporanic acid benzhydryl ester.

B. 7-Azido7-chloroformylcephalosporanic acid benzhydryl ester

7-Azido-7carboxycephalosporanic acid benzhydryl ester (4.0 g) was added to 15 ml thionyl chloride and the mixture was stirred in an ice bath for 1 h. The excess thionyl chloride was removed in vacuo and the residue was washed with dry acetone; 7-Azido-7-chloroformylcephalosporanic acid ester.

C. 7-Azido7-carbomethoxycephalosporanoic acid benzhydryl ester

7-Azido-7chloroformylcephalosporanic acid benzhydryl ester (2.63 g, 0.005) was dissolved in methylene chloride 15 ml at 5 ° C. Pyridine 40 mg and methanol 1.0 ml were added slowly. The solvent was removed in vacuo and the product was isolated by chromatography on silica gel.

D. 7-Amin7-carbomethoxy cephalosporic acid new benzhydryl ester

1.0 g of 7-azido-7-carbomethoxycephalosporanoic acid benzhydryl ester was dissolved in 100 ml of dioxane. 1.0 g of platinum oxide is added and the reaction mixture is stirred under hydrogen at atmospheric pressure for 1 h. Another amount of 1.0 g of platinum oxide is added and the reaction mixture is again placed under hydrogen and stirred for 3 h until the azide reacts completely ascertained by IR analysis of aliquots. The solvent was removed under reduced pressure and the residue was mixed with 50 ml of chloroform and filtered through silica gel G into chloroform in a 60 ml sintered glass funnel. The material was eluted with chloroform until 200 ml of chloroform was collected. The chloroform was removed under reduced pressure to give 0.632 g of 7-amino-7-carbomethoxycephalosporanic acid benzhydryl ester, which was acylated directly without further purification.

E. 7-Carbomethoxy-7- (2-thienylacetamide) -cephalosporanic acid benzhydryl ester. 0.632 g of 7-amine-7-carbomethoxycephalosporanoic acid benzhydryl ester was mixed with 25 ml of methylene chloride and cooled to 0 ° C. 0.6 ml, 0.038 mol of 2-thienylacetyl chloride were added dropwise over a period of 30 sec, followed by 0.6 ml, 0.01 mol of pyridine 60 sec later. The reaction mixture was stirred at 0 ° C for 15 min and poured onto crushed ice. The mixture was stirred and the organic layer was separated and washed once with 20 ml of water, once with 20 ml of 5% sodium bicarbonate solution and once again with 20 ml of water. The methylene chloride was dried and evaporated to dryness to give 1.417 g of crude product. This material is applied to a column of 60 g silica gel in benzene and the column is washed with benzene, 100 ml fractions are collected, followed by 300 ml in three fractions of methylene chloride / benzene / 1: 1 /, and 500 ml of methylene chloride in 5 fractions . The product was removed from the column by elution with 400 ml of chloroform in 4 fractions to give 0.592 g. This material was stirred in 25 ml of methylene chloride and stirred at room temperature with 20 ml of a solution of 0.120 ml of sodium bicarbonate in water for 1/2 hour. The layers were separated and the organic layer was washed with water, dried and evaporated. to dryness, 0.420 g of 7-carbomethoxy-7- (2-thienylacetamido / cephalosporanic acid benzhydryl ester) is obtained.

F. 7-Carbomethoxy-7- (2-thienylacetamido / cephalosporic acid) sodium salt

0.420 g of 7carboxy-7- (2-thienylacetamido / cephalosporanic acid benzhydryl ester was dissolved in 3.5 ml of anisole and treated with 10 ml of trifluoroacetic acid at room temperature for 10 minutes. The trifluoroacetic acid and anisole were removed under reduced pressure. , a temperature of less than 40 ° C was maintained and the residue was stirred in 25 ml of chloroform and treated with 20 ml of water containing 0.120 g of sodium bicarbonate. The mixture was stirred for 1/2 h at room temperature and the organic phase was separated and washed with water. The combined aqueous phase was separated and washed with water. The combined aqueous phase was washed twice with methylene chloride and lyophilized to give 0.382 g of the 7-carbomethoxy-7/2-thienylacetamido / cephalosporanic acid sodium salt as a brownish solid.

Example 110

3-Pyridinmethyl-7-carbomethoxy7- (2-thienylacetamido / decephalosporanoic acid)

1.3 g of the 7-carbomethoxy-7- (2-thienylacetamido / cephalosporanic acid) sodium salt is dissolved in 3 ml of buffer solution prepared by dissolving 30 g of potassium iodide in 20 ml of water containing 5 ml of pyridine, bringing the pH to 6.5 with dilute acetic acid. The resulting reaction mixture was stirred at 80 ° C for 2 h and then cooled to room temperature. The solution was then freeze-dried, dissolved in 40 ml of 25% acetic acid, filtered, and the filtered solution was electrophoresed to give 3-pyridinmethyl-7-carbomethoxy-7- (2-thienylacetamido) -decephalosporanova acid.

Example 111

7-Dimethylcarboxamido-7- (thiophene-2-acetamido) -cephalosporic acid sodium salt

In the procedure described in Example 107 C, 7-azido-7-chloroformylcephalosporanic acid benzhydryl ester is reacted with diamethylamine instead of methanol to give the corresponding 7-dimethylcarboxamide compound, which is converted to the 7-dimethylcarboxamido-2-thioxamido-7-dimethylcarboxamido-2- acetamido / cephalosporanic acid using the procedure described in Example 109.

Example 112

7-Hydrazinocarboxyl-7- (2-furylacetamido / cephalosporic acid) sodium salt

When the 7-azide-7-chloroformylcephalosporanic acid benzyl ester was reacted with hydrazine instead of methanol according to the procedure described in Example 109C, the corresponding 7-hydrazincarboxycephalosporanate was obtained. This product is reduced to the corresponding 7-amino compound as described in Example 109 E, and the resulting 7-amino-7-hydrazincarboxylic-cephalosporanic acid benzhydryl ester is reacted with furylacetyl chloride as described in Example 107 E to give the benzhydryl ester (2-furylacetamido) -7-hydrazinecarboxylcephalosporanic acid. This product was converted to the sodium salt of 7-hydrazinecarboxyl-7- (2-furylacetamido / cephalosporanic acid using the procedure described in Example 109.

Example 113

7- (2-furylacetamide) -7-thiocarboxymethylcephalosporanic acid sodium salt

When 7azide-7-bromocephalosporanic acid benzhydryl ester was reacted with carbon disulfide instead of carbon dioxide as described in Example 109 A, 7-azide-7-dithiocarboxycephalosporanic acid benzhydryl ester was obtained. This product was converted to the 7- (2-furylacetamido) -7-thiocarboxymethylcephalosporanoic acid sodium salt using the procedure described in Examples 107-110.

Example 114

7a-Formyl-7 / 2thienylacetamido / cephalosporic acid sodium salt

Phosphoric acid was added to a solution of 200 mg of 7a-hydroxymethyl-7- (2-thienylacetamido / cephalosporanic acid sodium salt to pH 2-3. The solution was extracted immediately with ethyl acetate in three portions. The combined organic layers were washed sequentially with water and with saturated sodium chloride solution and finally dried over anhydrous magnesium sulfate. Concentration of the solution under reduced pressure at or below room temperature yielded 7a-hydroxy-7- (2-thienylacetamide / cephalosporic acid (158mg) in the form of a gum which solidified to a glassy mass. A solution of this material in 4 ml of a 1: 1 mixture of methylene chloride and alcohol-free chloroform was added at one stirring at room temperature, with strong stirring, to a solution prepared by the addition of 225 mg of anhydrous chromium trioxide to a mixture of 0.375 ml of pyridine and 6 ml of methylene chloride. After 5 min of stirring, the reaction mixture was diluted with 15 ml of ethyl acetate and treated with 4 ml of 2% hydrochloric acid. The phases are mixed well, filtered under pressure through an infuser. The phases were separated and the aqueous layer was extracted with ethyl acetate. The combined organic layers were washed with water and dried over anhydrous magnesium sulfate. Concentration under reduced pressure at room temperature gave 82 mg of 7a-formyl 7- (2-thienylacetamido / cephalosporic acid) as a non-crystalline foam. The solid sodium salt is obtained by dissolving the product in one equivalent of an aqueous solution of sodium hydroxide, followed by removal of water by lyophilization.

Example 115

7a-Carboxy-7- (2-thienylacetamido / cephalosporic acid) sodium salt

Silver oxide is prepared by adding 10% potassium hydroxide to an aqueous solution of equimolar amounts of silver nitrate and potassium permanganate. The precipitated silver (II) oxide was collected by filtration and washed with water to release the base and the permanganate.

To a suspension of 250 mg of silver (II) oxide in 1.5 ml of tetrahydrofuran-water 4: 1 was added a solution of 100 mg of 7a-formyl-7- (2-thienylacetamido / cephalosporanic acid in 1.5 ml of tetrahydrofuran. The mixture was stirred at room temperature until thin-layer chromatography analysis indicated that the formyl compound was fully reacted. The reaction mixture was filtered and the filtrate was treated with 2% hydrochloric acid to pH 2. Ethyl acetate and water were added to the acidified filtrate and mixed. The separated organic layer was washed, dried and concentrated under reduced pressure at room temperature. The resulting free acid was converted to the sodium salt as in Example 114.

Example 116

7-Methyl-7- (2-thienylacetamide / cephalosporic acid) sodium salt

A. 7-Dimethylboro-7-methylcephalosporanoic acid benzhydryl ester

A solution of 0.228 g of 7-diazocephalosporanic acid benzhydryl ester in 5 ml of methylene chloride, prepared as described above, was diluted to about 10 ml with tetrahydrofuran and cooled to about -78 ° C in a dry ice bath. To this cooled solution was added 3.5 ml of triethyl boronic solution with stirring for 10 min. The resulting reaction mixture was stirred for a further 10 min at -78 ° C. The infrared spectrum of a small aliquot of this solution at this point shows no diazo. The reaction mixture containing 7-dimethylboro-7-methylcephalosporanic acid benzhydryl ester was allowed to warm to room temperature. Evaporation of the solvent gives the product a solid form.

The trimethylborane solution used in this example was prepared as follows. Methyl magnesium iodide of 1.44 g of magnesium and 8.5 g of methyl iodide in 40 ml of ether is prepared and the resulting Grignard solution is diluted to 50 ml with additional ether. Place 5 ml of this solution in a three-necked round-bottom flask fitted with a drop funnel, magnetic stirrer and a tube with a nitrogen valve. The flask was connected to the tap of a gas tube fitted with a two-necked flask containing 5 ml of ether, cooled to ~ 78 ° C in a dry ice bath. The two flasks were placed under a nitrogen atmosphere, with nitrogen gas passing into the first flask and bubbling in the ether from the second flask. To the first flask was added dropwise 0.2 ml of boron trifluoride - ether diluted to 2 ml with ether, the resulting trimethyl boron collected in the second flask. The nitrogen stream was maintained until most of the ether in the first flask was purged into the second flask to give about 9 ml of trimethylboro solution.

B. 7-Azido7-methylcephalosporanic acid benzhydryl ester

To a solution of 7-dimethylboro-7-methylcephalosporanic acid benzhydryl ester in methylene chloride prepared as described in Example A was added a solution of bromazide in methylene chloride prepared as described below. The resulting reaction mixture turned orange and stirred at room temperature overnight. It is then poured into a cold solution of N / 10 sodium thiosulfate. The organic phase was separated, washed with an equal amount of aqueous 5% sodium bicarbonate solution and then with water. The washed organic phase was then dried over sodium sulfate and evaporated to give 7-azide-7-methylcephalosporanic acid benzhydryl ester.

The bromazide solution is prepared as follows. To a solution of 2.4 g of sodium azide in 4 ml of water was added 40 ml of methylene chloride and the mixture was cooled to 0 ° C. To this cooled solution was added dropwise 4 ml of a 50% aqueous sulfuric acid solution for 5 min and the mixture was stirred for another 5 min. The aqueous solution was frozen by cooling in a solid carbon dioxide bath and the organic phase was decanted and dried over anhydrous sodium sulfate. To the hydrochloric acid solution was added 2 g of N-bromosuccinamide and the mixture was stirred for 15 min at 0 ° C until the N-bromosuccinamide was dissolved.

C. 7-Amin7-methylcephalosporanic acid benzhydryl ester

1.0 g of 7-azido-7-methyl-cephalosporanic acid benzhydryl ester was dissolved in 100 ml of dioxane. 1.0 g of platinum oxide is added, the mixture is stirred under nitrogen at atmospheric pressure for 1 h. Another 1 g of platinum oxide is added and the reaction mixture is again brought under nitrogen and stirred for 3 h until the azide is fully reacted, which is determined by infrared analysis of an aliquot. The solvent was removed under reduced pressure and the residue was stirred in 50 ml of chloroform and filtered through silica gel G into chloroform in a 60 ml sintered glass shot filter. The material was eluted with chloroform until 200 ml of chloroform was collected. The latter was removed under reduced pressure to give 0.632 g of 7-amino-7-methylcephalosporanic acid benzhydryl ester, which was acylated directly without further purification.

D. 7-Methyl7- (2-thienylacetamido / cephalosporic acid benzhydryl ester

0.623 g of methyl-7-aminocephalosporanic acid benzhydryl ester was stirred in 25 ml of methylene chloride and cooled to 0 ° C. 0.6 ml / 0.038 mol / 2-thienylacetyl chloride was added dropwise over 30 seconds, followed by 0.6 ml / 0.01 mol / pyridine 60 seconds later. The reaction mixture was stirred at 0 ° C for 15 min and poured into crushed ice. The mixture was shaken and the organic layer separated and washed once with 20 ml of water, once with 20 ml of 5% sodium bicarbonate and once more with 20 ml of water. The methylene84 chloride was dried and evaporated to dryness to give 1.417 g of crude product. This material was placed in a column of 60 g silica gel with benzene and the column eluted with benzene, 100 ml fractions were then collected, then 300 ml in three fractions with methylene chloride / benzene (1: 1) and 500 ml of methylene chloride in 5 fractions. The product was removed from the column by elution with 400 ml of chloroform in 4 fractions to give 0.592 g. This material was stirred in 25 ml of methylene chloride and stirred for 1/2 hour at room temperature with 20 ml of a solution of 0.120 g of sodium bicarbonate in water. The layers were separated and the organic layer was washed with water, dried and evaporated to dryness to give 0.420 g of 7-methyl-7- (2-thienylacetamido / cephalosporic acid benzhydryl ester).

E. 7-Methyl-7- (2-thienylacetamido / cephalosporanoic acid) sodium salt

0.420 g of 7-methyl-7- (2-thienylacetamido / cephalosporanic acid benzhydryl ester is dissolved in

3.5 ml of anisole and treated with 10 ml of trifluoroacetic acid at room temperature for 10 minutes. Trifluoroacetic acid and anisole were removed under reduced pressure, the temperature was maintained below 40 ° C and the residue was dissolved in chloroform and treated with 20 ml of water containing 0,120 g of sodium bicarbonate. The mixture was stirred for 1/2 h at room temperature and the organic phase was separated and washed with water. The combined aqueous phase was washed twice with methylene chloride and lyophilized to give 0.382 g of the 7-methyl-7- (2-thienylacetamido / cephalosporanic acid sodium salt) as a brownish solid.

Example 117

3-Pyridinmethyl-7-methyl-7- (2-thienylacetamido / decephalosporanoic acid)

1.3 g of 7-methyl-7/2-thienylacetamido / cephalosporanic acid sodium salt was dissolved in 3 ml of buffer solution prepared by dissolving 30 g of potassium iodide in 20 ml of water containing 5 ml of pyridine and adjusting the pH of 6.5 with dilute acetic acid. The resulting reaction mixture was stirred at 80 ° C for 2 h and then cooled to room temperature. The solution was freeze-dried, dissolved in 40 ml of 25% acetic acid, filtered and the filtrate solution electrophoresed to give 3-pyridinmethyl-7-methyl / 2-thienylacetamido / decephalosporic acid.

Example 118

A. 7-Difluoromethylenetsephalosporanic acid benzhydryl ester

7-Diazocephalosporanic acid benzhydryl ester (4.5 g, 0.01 mol) in 10 ml of dry dioxane was added to a solution of 4.1 g of 0.05 mol of thiocarbonyl fluoride and allowed to stand at room temperature until the reddish color disappears. the diazo compound. The reaction mixture was refluxed for 45 min and the solvent removed in vacuo. The product was chromatographed on silica gel using chloroform: methanol as eluent to give 7-difluoromethylencephalosporanic acid benzhydryl ester.

B. 7-Difluoromethyl l-7-bromocephalospora new acid benzhydryl ester

7-Difluoromethylenetsephalosporanic acid benzhydryl ester

2.35 g, 0.005 mol and finely saturated silver monofluoride 1.90 g, 0.015 mol in 50 ml of benzene are vigorously stirred. 0.80 g of bromine was added dropwise,

0.005 mol. 7 Trifluoromethyl-7-bromocephalosporanic acid benzhydryl ester was isolated by chromatography on silica gel.

C. 7-Trifluoromethyl-7-aminocephalosporanoic acid benzhydryl ester

7-Trifluoromethyl-7-bromocephalosporanic acid benzhydryl ester 1.14 g, 0.02 mol was dissolved in 20 ml of dry ether and cooled to -20 ° C. To this cooled solution was slowly added 0.4 ml of 5 M ether methyl lithium solution with vigorous stirring. After 1/2 h, 66 mg of o-methylhydroxylamine was added. The mixture was kept at -10 ° C for 1/2 h and allowed to warm to room temperature before refluxing for 2 h. The ether was removed in vacuo and the 7-trifluoromethyl7-aminocephalosporanoic acid benzhydryl ester was isolated by chromatography on silica gel.

D. 7- (2-Thiophenacetamido) -7-trifluoromethylcephalosporanoic acid benzhydryl ester

7-Trifluoromethyl-7-aminocephalosporanic acid benzhydryl ester, prepared as described in C, is acylated by reaction with 2-thiophenacetyl chloride using the method described in the example to obtain 7- (2-thiophenacetate benzhydryl ester) N-7-Trifluoromethylcephalosporanic acid.

E. 7- (2-Thiophenacetamido) -7-trifluoromethylcephalosporanoic acid sodium salt

7- (2-Thiophenacetamido) -7-trifluoromethylcephalosporanic acid benzhydryl ester was dissolved in 3.5 ml of anisole and treated with 10 ml of trifluoroacetic acid at room temperature for 10 minutes. The trifluoroacetic acid and the anisole were removed under reduced pressure. kept below 40 ° C and the residue was stirred in 25 ml of chloroform and treated with 20 ml of water containing 0.120 g of sodium bicarbonate. The mixture was stirred for 1/2 h at room temperature and the organic phase was separated and washed with water. The combined aqueous phases were washed twice with methylene chloride and lyophilized to give 0.382 g of the 7- (2-thiophenacetamido) -7-trifluoromethylcephalosporanoic acid sodium salt.

Example 119

A. 7-Ethynyl-7-azidecephalosporanic acid benzhydryl ester

A stirred solution of 0.900 g of 7-diazosephalosporanic acid benzhydryl ester in 10 ml of ether and in 10 ml of methylene chloride was cooled to -78 ° C under nitrogen atmosphere and treated dropwise with a solution of trietinyl boron. The addition was stopped periodically and the progress of the reaction was detected by infrared analysis of small aliquots. When the diazo compound was fully reacted, the remaining solution of triethyl boron was removed. Bromazide in methylene chloride, 25 ml of 0.28 N solution was then added over a period of 20 minutes. The cooling bath was removed and the mixture was stirred for a further 60 min at room temperature.

The reaction mixture was poured onto a solution of 20 ml of 0.1 N sodium thiosulphate and 20 ml of 0.5 M phosphate buffer with pH 7. The organic phase was separated, washed with water and saturated sodium chloride solution, dried over anhydrous magnesium sulfate. and evaporated in vacuo. The residue was chromatographed on silica gel using benzene as eluent to give 7α-ethynyl-7-azidocephalosporanoic benzhydryl ester and 7β-ετπΗΗ-azidocephalosporanic acid benzhydryl ester.

The trietinyl boron solution used above was prepared as follows. A solution of 5.42 g of borontrifluoride in 20 ml of ether was stirred at -78 ° C under nitrogen atmosphere, while 2.88 g of sodium acetylide in 20 ml of xylene were added over a period of 90 minutes. The reaction mixture was a white solid. suspended in a clear liquid, rapidly transferred to an absorber and filtered through a dry drip funnel equipped with an ice casing. The solution is then used immediately. In no case should the temperature of trietinylborane be allowed to exceed -60 ° C.

B. 7a-Ethynyl7-aminocephalosporanoic acid benzhydryl ester

To a stirred solution of 0.028 g of anhydrous cobalt II bromide in 20 ml of absolute ethanol was added 0.061 g of 2,2'-bipyridine. After all the bipyridine was dissolved, a solution of 0.315 g of 7a-ethynyl-7-azidocephalosporanic acid benzhydryl ester in 5 ml of absolute ethanol was added followed by 0.073 g of sodium borohydride. The reaction mixture was stirred for 15 min at room temperature and then cooled sharply by the addition of cold acetic acid. The mixture was diluted with 25 ml of water and extracted with three 20 ml portions of ether. The combined extracts were washed with phosphate buffer pH 7 and saturated sodium chloride solution and dried over anhydrous magnesium sulfate. Evaporation of the solvent under reduced pressure afforded the 7a-ethynyl7-aminocephalosporanic acid benzhydryl ester. This product is acylated without purification.

C. 7a-Ethynyl-7- (2-thienylacetamido) -cephalosporanoic acid benzhydryl ester

Ice-cooled, stirred solution of 0.274 g benzhydryl ester

7a-ethynyl-7-aminocephalosporanic acid in 10 ml of methylene chloride was treated with 0.25 ml of 2-thienylacetyl chloride for 30 sec and after 1 min with 0.25 ml of pyridine. The reaction mixture was stirred for a further 15 min at 0 ° C and then poured onto crushed ice. The methylene chloride phase was separated, washed with 10 ml of water, 2 x 40 ml of phosphate buffer with pH 2, 10 ml of water and dried over anhydrous magnesium sulfate. Evaporation of the solvent left a yellow gum which was chromatographed on 15 g of silica gel. The column was eluted with 75 ml of benzene, 150 ml of 1: 1 benzene methylene chloride, 150 ml of methylene chloride and 300 ml of chloroform. Evaporation of the chloroform fraction under reduced pressure gave an oil which was mixed with 10 ml of methylene chloride and stirred for 30 minutes with a solution of 0.050 g of sodium bicarbonate in 5 ml of water. The methylene chloride phase was separated, washed with 5 ml of water, dried over anhydrous magnesium sulphate and evaporated to give 7a-ethynyl-7- (2-thienylacetamido) -cephalic-sporic acid benzhydryl ester.

D. 7a-Ethynyl-7 (2-thienylacetamido) -cephalosporanic acid sodium salt

Benzhydryl 7a-ethynyl-7- (2-thienylacetamido) -cephalosporanoic acid 0.179 g was dissolved in 1.5 ml of anisole and treated with 4.5 ml of trifluoroacetic acid. After keeping at room temperature for 10 minutes, the solution was evaporated in vacuo to remove excess reagents. The residue was dissolved in 10 ml of methylene chloride and stirred for 30 minutes with a solution of 0.045 g of sodium bicarbonate in 7.5 ml of water. The phases are separated; The organic layer was extracted with 4 ml of water, the combined aqueous solutions were washed with 3 x 4 ml of methylene chloride and lyophilized to give a powder. The crude product was partially dissolved in anhydrous methanol and filtered. Evaporation of the solvent from the filtrate afforded the 7α-ethynyl-7- (2-thienylacetamido) -cephalosporanic acid sodium salt.

In the procedure described in steps B, C and D, the 7β-ethynyl-7-azidocephalosporanic acid benzhydryl ester can be converted to the 7β-ethynyl-7- (2-thienylacetamido) -cephalosporanic acid sodium salt. The acetoxymethyl group of this compound and the 7β-ethynyl compound can be converted to another 3-substituted cephalosporin as described in the above examples. The acetoxy group may be replaced by nitrogen bases and other compounds of greater nucleophilicity than the oxygen nucleophile. Other 3-substituted compounds such as 3-hydroxymethyl may be prepared and then modified to give other novel compounds. The corresponding 3-desacetyl derivatives can also be prepared using the 7-diazodetaxoxycephalosporanic acid benzhydryl ester in step A described above. Using the procedures described in other examples, other 7-acylamide compounds can be obtained.

Example 120

7a-Phenyl-7- (2- (4-pyridylthio) acetamido] -cephalosporic acid.

A. 7-Phenyl7-azidocephalosporanoic acid benzhydryl ester

With stirring, a solution of 1,350 g of 7-diazocephalosporanic acid benzhydryl ester in methylene chloride and ether of 15 ml was cooled to -78 ° C under a nitrogen atmosphere and treated dropwise over a period of 20 minutes with a solution of 0.727 g of triphenylboro in 20 ml of ether . The mixture was stirred for a further 10 min at -78 &lt; ^{9 &gt}; C, during which time the diazo compound was fully reacted, which was established by IR analysis of small aliquots. Bromazide in methylene chloride, 45 ml of 0.28 N solution was added over a period of 30 minutes. The cooling bath was removed and the mixture was stirred for 2 h at room temperature. The reaction mixture was poured into a solution of 30 ml of 0.1 N thiosulphate and 30 ml of 0.5 M phosphate buffer and pH 7 and stirred. The organic phase was separated, washed with water and saturated sodium chloride solution, dried over anhydrous magnesium sulfate and evaporated. The crude product was chromatographed on silica gel using benzene as eluent to give 7α-phenyl-7-azidocephalosporanic acid benzhydryl ester and 7β-phenyl-7-azidocetalacopopanoic acid benzhydryl ester.

B. 7a-Phenyl-7-aminocephalosporanoic acid benzhydryl ester

A mixture of 0.450 g of 7a-phenyl-7-azidocephalosporanic acid benzhydryl ester and 0.450 g of platinum oxide in 45 ml of dioxane was stirred under hydrogen at atmospheric pressure for 1 h. After purging the system with nitrogen, another 0.450 g of platinum oxide were added and the reaction was The mixture was again placed under hydrogen and stirred for 2 h. At this temperature, the azide reacted completely as evidenced by IR analysis of aliquots. The solvent was evaporated under reduced pressure and the residue was taken up in 25 ml of chloroform and filtered through a pad of silica gel G and diatomaceous earth in a 60 ml sintered glass funnel. The product was eluted with chloroform until 200 ml of solution was collected. Evaporation of the solvent under reduced pressure afforded the 7a-phenyl-7-aminocephalosporic acid benzhydryl ester, which was acylated directly without purification.

C. 7a-Phenyl- [2- (4-pyridylthio) -acetamide] cephalosporic acid benzhydryl ester.

To a stirred solution of 0.395 g of 7α-phenyl-7-aminocephalosporanic acid benzhydryl ester in 15 ml of dry methylene chloride was added 0.130 g (4-pyridylthio) -acetic acid and then 5 ml of dimethylformamide to give a solution. Dicyclohexylcarbodiimide 0.159 g was then added and the mixture was stirred at room temperature and under a nitrogen atmosphere for 18 'h. The dicyclohexylurea precipitate was removed by filtration and the filtrate was evaporated under high vacuum. The residue was dissolved in 25 ml of chloroform and washed with water, 2% sodium bicarbonate, water and saturated sodium chloride solution. The chloroform was dried over anhydrous magnesium sulfate and evaporated under reduced pressure to give the crude product. The latter was chromatographed on 20 g of silica gel using ethyl acetate in a benzene gradient amount as eluent to give 7a-phenyl-7- (2- (4-pyridylthio) -acetamide] -cephalosporic acid benzhydryl ester.

D. 7a-Phenyl-7- (2- (4-pyridylthio) -acetamido] -cephalosporic acid.

To a stirred solution of 0.330 g of 7a-phenyl-7 / 2- (4-pyridylthio) -acetamide) -cephalosporanoic acid benzhydryl ester in 2.5 ml of anisole was added 7 ml of trifluoroacetic acid. The reaction mixture was stirred for 10 min at room temperature. The excess anisole and trifluoroacetic acid was removed under high vacuum and the residue was mixed with 20 ml of chloroform and covered with ml of water containing 0.100 g of sodium bicarbonate. The mixture was stirred for 30 min at room temperature and the organic phase was separated and washed with water. The combined aqueous phases were washed with methylene chloride, covered with ethyl acetate and acidified to pH 3 with 2 N hydrochloric acid. The ethyl acetate phase was separated, dried over anhydrous sodium sulfate and evaporated to give 7a-phenyl-7- (2- (4-pyridylthio) -acetamide) -cephalosporic acid.

In the procedures described in steps B-D, the 7α-phenyl-7-azidocephalosporanoic acid benzhydryl ester is converted to 7β-phenyl-7- (2- (4-pyridylthio) -acetamide / cephalosporanoic acid.

The acetoxymethyl group of both 7a- and 7β-phenyl-7- (2- (4-piperidylthio) -acetamide / cephalosporic acids can be converted to other 3-substituted products using the procedures of the above examples. The acetoxy group may be replaced by nitrogen bases and other compounds of greater nucleophilicity than the oxygen nucleophile. 3-Acetoxymethyl can be catalytically reduced in the presence of hydrogen to give a 3-methyl derivative. Other 3-substituted compounds can also be prepared, for example 3-hydroxymethyl and converted to other useful compounds. Also, 7α-phenyl-7-aminocephalosporanic acid benzhydryl ester can be acylated with other acylating agents, such as phenylacetyl chloride, thienylacetyl chloride, furylacetyl chloride and the like, to obtain the corresponding 7-acylamide compounds, below.

Example 121

7a-ethyl-7- (O-2-amino-α-cephalosporic acid)

A. 7-Ethyl-7-azidocephalosporanic acid benzhydryl ester

A solution of 1,350 g of 7-diazocephalosporanic acid benzhydryl ester in 15 ml of methylene chloride and ether was cooled to ~ 78 ° C under nitrogen atmosphere and stirred while adding 0.295 g of triethyl boron in 15 ml of ether dropwise over 15 min. The mixture was stirred for a further 5 min at -78 ° C, during which time the diazo compound was fully reacted, which was established by IR analysis of small aliquots. Then bromazide in methylene chloride, 45 ml of 0.28 N solution was added dropwise over a period of 30 minutes. The cooling bath was removed and the mixture was stirred for 2 hours at room temperature. The reaction mixture was poured into a solution of 30 ml of 0.1 N sodium thiosulfate and 0.5 M phosphate buffer at pH 7 and stirred. The organic phase was separated, washed with water, saturated sodium chloride solution, dried over anhydrous magnesium sulfate and evaporated. The residue was chromatographed on silica gel using benzene as eluent to give 7a-ethyl-7-azido-cephalosporanate benzhydryl ester and 73-ethyl-7-azidocephalosporanic benzhydryl.

B. 7a-ethyl7-aminocephalosporanoic acid benzhydryl ester

A solution of 0.520 g of 7a-ethyl-7-azidocephalosporanic acid benzhydryl ester in 150 ml of dioxane was treated with 0.520 g of platinum oxide and the mixture was stirred under hydrogen at atmospheric pressure for one hour. After purging the mixture with nitrogen, an additional 0.520 g of platinum oxide was added. The mixture was then moved under hydrogen and stirred for 2 h, during which time the azide reacted completely, as determined by IR analysis of aliquots. The dioxane was removed under reduced pressure and the residue was mixed with 30 ml of chloroform and filtered through a pad of silica gel G and diatomaceous earth in a 60 ml sintered glass funnel. The product was eluted with chloroform until 200 ml of solution was collected. Evaporation of the solvent under reduced pressure affords 7a-ethyl7-aminocephalosporanoic acid benzhydryl ester

C. 7a-ethyl7- (O-2-azido-2-phenylacetamide) -cephalosporanic acid benzhydryl ester. To a stirred solution of 0.418 g of 7a-ethyl-7-aminocephalosporanoic acid 7α-ethyl-7-aminocephalosporanic acid in 0 ml of C was rapidly added 0.350 g of D-a-azidophenylacetic acid and then 0.30 ml of pyridine 60 sec. late. The reaction mixture was stirred for 30 min at 0 ° C and poured onto 40 ml of water. The mixture was stirred and the methylene chloride phase was separated and washed sequentially with water, dilute sodium bicarbonate, water and saturated sodium chloride solution. The methylene chloride was removed under reduced pressure after drying over anhydrous magnesium sulfate to give the crude product. This product was chromatographed on 40 g of silica gel using chloroform in benzene in a gradient amount as eluent to give 7aethyl-7 / O-2-azido-2-phenylacetamide / cephalosporic acid benzhydryl ester.

D. 7a-ethyl-7 (N-2-azido-2-phenylacetamide) -cephalosporic acid. A solution of 0.388 g of 7a-ethyl-7 / O-2-azido-2-phenylacetamido / -cephalosporanic acid benzhydryl ester in 1.0 ml of anisole is cooled in an ice bath and treated with 4.0 ml of trifluoroacetic acid. The reaction mixture was kept at 0 ° C for 10 min with rotation from time to time, then evaporated in vacuo to remove excess anisole and trifluoroacetic acid. The residue was mixed with 20 ml of methylene chloride and extracted with three 5 ml portions of saturated sodium bicarbonate solution. The combined aqueous extracts were washed with two 5 ml portions of methylene chloride, acidified to pH 2 with 2.5 N hydrochloric acid and extracted with 4 portions of 10 ml ethyl acetate. The organic extracts were combined and dried over anhydrous magnesium sulfate. Evaporation of the solvent afforded 7a-ethyl-7 / O-2-azido-2-phenylacetamido / cephalosporic acid.

E. 7a-ethyl-7 (O-2-amino-2-phenylacetamido) -cephalosporanic acid. 0.291 g of 7a-ethyl-7 / O-2-azido-2-phenylacetamido / cephalosporanic acid are mixed with 3.0 ml of acetic acid and 4.5 ml of water, cooled in an ice bath and treated with 1.550g of zinc powder . The mixture was stirred for 10 min and then filtered through sintered glass. The zinc is washed with 20 ml of water. The filtrate and washes were combined, saturated with hydrogen sulfide and filtered through a layer of diatomaceous earth. Lyophilization of the filtrate afforded the crude product 7a-ethyl7 / N-2-amino-2-phenylacetamido / cephalosporic acid. The product is released from acetic acid by re-dissolving in water and lyophilizing.

Similarly, 7β-ethyl-7-azidosephalosporanate can be converted to 7β-ethyl-7 / D-2-amino-2-phenylacetamido / -cephalosporanic acid according to procedures C-E. The use of L-α-azidophenylacetic acid and DL-α-azidophenylacetic acid according to step C and the subsequent transformations gave 7α-ethyl-7- (L-2-amino-2-phenylacetates to β-cephalosporic acid, 7β-ethyl-7- / β-2 -Amino-2-phenylacetamido] -cephalosporanoic acid, 7aethyl-7- (N-2-amino-2-phenylacetamido) -cephalosporanoic acid and 7βethyl-7- (OB-2-amino-2-phenylacetamido) -cephalosporic acid. Using the other acylating agents in Example C above, analogous 7-acylamido compounds were obtained.

The acetoxymethyl group of these compounds can be converted to other 3-substituted compounds in the usual way. The acetoxy group may be displaced by nitrogen bases and other compounds with a higher nucleophilicity than the oxygen nucleophile. Other 3-substituted compounds, for example 3-hydroxymethyl, may also be prepared and converted to other derivatives. In addition, the 3-acetoxymethyl group can be catalytically reduced in the presence of hydrogen to give 3-methylcephalosporins.

Example 122

A. 7-Azidocephalosporanic acid benzhydryl ester

To a solution of 0.900 g, 0.002 mol of 7-diazocephalosporanic acid benzhydryl ester in 10 ml of CH ₂ Cl ₂ and 10 ml of ether, cooled to -78 ° C under a nitrogen atmosphere, was added dropwise over 10 minutes 10 ml of 0, 02 M solution of tribenzylboron in ether.

When aliquot control portions showed the absence of diazo compound by IR analysis, the reaction mixture was stirred for a further 5 min at -78 ° C and then treated with 40 ml of a 0.3 N solution of BrN ₃ in methylene chloride. The reaction mixture was stirred at -78 ° C for 15 min and then allowed to reach room temperature for 45 min and stirred for another 20 min at room temperature. The reaction mixture was poured into ice-cold 5% en NaHCO _{3 solution} , stirred and the organic phase separated and washed once with N / 10 thiosulphate 30 ml, then with water, dried and evaporated. The residue was chromatographed on 30 g of silica gel to give 7α-benzyl-7-azidocephalosporanic acid benzhydryl ester and 7β-66Η3ΗΑ-7-33ΗΛου, εφΒlosporic acid benzhydryl ester.

B. 7a-Benzyl-7-aminocephalosporanoic acid benzhydryl ester

7,500 g of 7azido-7α-cephalosporanic acid benzhydryl ester was dissolved in 25 ml of ethyl acetate and 0,500 g of platinum oxide was added and the mixture was stirred overnight under hydrogen. The catalyst was filtered off, the filtrate was evaporated and the residue was chromatographed on silica gel 15 g to give the product.

C. 7a-Benzyl-7- (p-carboxymethylphenylacetamido) -cephalosporanic acid benzhydryl ester. 0,400 g of 7a-benzyl-7-aminocephalosporanic acid benzhydryl ester was dissolved in 25 ml of CH ₂ Cl ₂ and cooled to 0 ° C. 0.400 g of p-phenylenediacetyl chloride was added followed by 0.400 g of pyridine after 60 sec. The reaction mixture was shaken, the organic phase separated and washed once with pH 2 phosphate buffer and then with water. The organic phase was dried and evaporated. The crude product thus obtained is purified by chromatography on silica gel.

D. 7a-Benzyl7- (p-carboxymethylphenylacetamido / cephalosporic acid disodium salt)

0.325 g of 7abenzyl-7- (p-carboxymethylphenyl) benzhydryl ester cetane / cephal was challenged with new acid and dissolved in 3 ml of anisole and treated with 10 ml of trifluoroacetic acid at room temperature for 10 minutes. Trifluoroacetic acid and anisole evaporated under reduced pressure at 40 ° C. The residue was taken up in 25 ml of ethyl acetate, purified by acetic acid and treated with 20 ml of H ₂ O containing 0.120 g of sodium bicarbonate. The mixture was stirred for 1/2 h and the organic phase was separated and washed with water. The combined aqueous phase was washed twice with ethyl acetate and then freeze-dried to give a product which was further purified by recrystallization from methanol / isopropanol.

Interaction of the 7β-benzyl-7-azidecalocoranoic acid benzhydryl ester in the above sequence gives 7β-6εΗ3ΠΑ-7- / ρ-κ3ρboxymethylphenylacetamido / cephalosporanate.

When another acylating agent such as phenylacetyl chloride, 2-thienylacetyl chloride, 2-furylacetyl chloride is used in step C, the corresponding 7-acylamido compounds are prepared according to the procedures of the examples above. The resulting cephalosporins are converted to other 3-substituted products, such as 3-methyl, 3-hydroxymethyl, and 3-pyridinmethyl compounds, according to methods well known in the art described in the foregoing examples.

Example 123

7-α-vinyl-7- (phenylacetamido) cephalosporic acid

A. 7-vinyl7-azidecephalosporanic acid benzhydryl ester

0.9 g of 0.002 mol of 7-diazocephalosporanic acid benzhydryl ester was dissolved in 10 ml of methylene chloride and 10 ml of ether, cooled to -78 ° C under nitrogen and treated dropwise over 15 min with 10 ml of 0.02 M solution. of trinylborne in ether. Checking a small aliquot portion showed92 ir in the absence of the diazo compound. The reaction mixture was stirred at -78 ° C for a further 5 min and then treated with 40 ml of a 0.3 N solution of bromazide in methylene chloride. The reaction mixture was stirred at -78 ° C for 15 min and allowed to reach room temperature after 45 min, then stirred for another 20 min at room temperature.

The reaction mixture was poured into ice-cold 5% sodium bicarbonate solution, shaken and the organic phase separated, washed once with 0.1 N sodium thiosulphate solution 30 ml and then with water, dried and evaporated. The residue was chromatographed on 30 g of silica gel to give 7-avinyl-7-azidcephalosporanic acid benzhydryl ester and 7-a-vinyl7-azidcephalosporanic acid benzhydryl ester.

B. 7-α-vinyl-7-aminocephalosporanoic acid benzhydryl ester

0.030 g of anhydrous cobalt bromide was dissolved in 50 ml of absolute ethanol and treated with 0.065 g of αα'-dipyridyl. 0.300 g of 7-a-vinyl-7-azidcephalosporanic acid benzhydryl ester are then added, dissolved in 2 ml of ethanol, followed by 0.080 g of sodium borohydride. After rapid gas evolution ceased for about 15 minutes, the reaction mixture was poured into phosphate buffer at pH 7 and extracted with methylene chloride, washed once with water, dried and evaporated. The residue was chromatographed on silica gel 10 g to give 7-a-vinyl-7-aminocephalosporanic acid benzhydryl ester.

C. 7-α-vinyl-7- (phenylacetamide) cephalosporic acid benzhydryl ester

0.450 g of 7α-vinyl-7-aminocephalosporanic acid benzhydryl ester was dissolved in 25 ml of methylene chloride and cooled to 0 ° C. 0.400 ml of phenylacetyl chloride, then 0.400 ml of pyridine are added. The reaction mixture was stirred for 25 min at 0 ° C and poured into crushed ice. The mixture was shaken, the organic phase separated and washed with phosphate buffer with pH 2 and then with water. The organic phase was dried and evaporated. The residue was chromatographed on silica gel to give a purified product.

D. 7-α-vinyl-7- (phenylacetamide / cephalosporic acid)

0.350 g of 7a-vinyl-7- (phenylacetamido) -cephalosporanic acid benzhydryl ester was dissolved in 3 ml of anisole and treated with 10 ml of trifluoroacetic acid at room temperature for 10 minutes. The trifluoroacetic acid and the anisole were evaporated under reduced pressure. below 40 ° C. The residue was mixed with methylene chloride 25 ml and stirred with 20 ml of water containing 0.120 g of sodium bicarbonate. The organic phase was separated and washed once with water. The combined aqueous phase was adjusted to pH 2 and extracted with ethyl acetate. The ethyl acetate extracts were dried and evaporated to give the crude product, which was purified by recrystallization from ethyl acetate / ether.

Interaction of 7-p-vinyl-7-azidcephalosporanic acid benzhydryl ester in the above sequence gives 7-3-vinyl-7- (phenylacetamido / cephalosporanic acid).

Example 124

A. 7-α-aminoethyl-7- (2-benzhydryloxycarbonylphenylacetamido) -cephalosporic acid benzhydryl ester

0,800 g of 7a-cyan-7- (2-benzhydroxycarbonylphenylacetamido) -cephalosporanoic acid benzhydryl ester obtained as described in Example 108 C was dissolved in 10 ml of tetrahydrofuran and cooled to 0 ° C. 1 ml of a 1M solution of BH ₃ THF in tetrahydrofuran was added dropwise over 5 min and the mixture was stirred at 0 ° C for 1/2 h.

The reaction mixture was poured into phosphate buffer pH 7 and extracted with methylene chloride. The methylene chloride extract was dried and evaporated to give a crude product which was purified by chromatography on silica gel.

B. 7-a-aminoethyl-7- / 2-carboxyphenylacetamide / cephalosporic acid disodium salt

0.450 g of 7a-aminoethyl-7- (2-benzhydryloxycarbonylphenylacetamido / cephalosporanoic acid benzhydryl ester was dissolved in 3 ml of anisole, 10 ml of trifluoroacetic acid was added and the reaction mixture was left at room temperature for 10 min. removed under reduced pressure below 40 ° C. The residue was taken up in 25 ml of ethyl acetate and treated with 15 ml of water containing 0,200 g of sodium bicarbonate. The mixture was stirred for 15 min. The organic phase was evaporated and washed once with water. The combined aqueous phase was washed once with methylene chloride and then freeze-dried. The resulting product was purified by recrystallization from methanol / isopropanol.

Reaction of 7-p-cyan-7- (2-benzhydryloxycarbonylphenylacetamido) -cephalosporic acid benzhydryl ester in the above sequence gives 7-p-aminomethyl-7- (2-carbonylphenylacetamide / cephalosporanate).

Example 125

7-Hydroxymethyl 7- (2-thienylacetamide) cephalosporanoic acid sodium salt

A. 7- (4-Nitrobenzylidinamino / cephalosporanic acid benzhydryl ester)

Equimolar mixture of bezhindrilov ester of 7-aminocephalosporanic acid and 4-nitrobenzaldehyde was heated under nitrogen in 200 ml of benzene per gram of aldehyde and water is removed azeotropically for 1 h. The solution was evaporated under reduced pressure to obtain a foam IR / CHC1 ₃ /, shows bands at 5.60 β-lactams and

5.75 m esters, while 60 Hz NMR peaks at 518, 516, (1H), 596, 587, 575, 566 (AB quartet, 4H), 439 (1H, 416 (1H), 330, 328, 325, 323 (doublet of doublets, 1H), 311, 306 (1H), 308, 295, 288, 274 (AB quartet (2H) and 119 (3H), the numbers are in Hz at an internal witness TMS in deuterochloroform.

Thin layer chromatography on 250 μg silica gel plates with 10% ethyl acetate in chloroform showed a single spot with R _f 0.58, only traces of the starting material can be detected.

B. 7-Hydroxymethyl-7/4-nitrobenzylidinamino / cephalosporic acid benzhydryl ester

A weak stream of nitrogen was passed through a glass ampoule containing 60 mg of 7- (4-nitrobenzylidinamino / cephalosporanic acid benzhydryl ester) and 0.3 ml of N, N-dimethylformamide was added after a few minutes. Nitrogen leakage was continued by bubbling through the green-brown solution for about 30 s and then flowing from formaldehyde gas to nitrogen, obtained by heating ~ 15 mg of paraformaldehyde in the nitrogen stream. The color was removed and the resulting solution was evaporated to a rubber mass in vacuo. The tire was purified by dissolving in a small volume of chloroform and recovering under high vacuum to a tire. The product exhibits a pure IR absorption spectrum for hydroxy, β-lactam and ester groups. The NMR spectrum in deuterium chloroform shows the expected singlet for the benzylidone proton and the new absorption associated with the hydroxymethyl group.

C. 7-Hydroxymethyl-7-aminocephalosporanoic acid benzhydryl ester, tosylate salt

A mixture of 100 mg of powdered 2,4-dinitrophenylhydrazine, 85.5 mg of p-toluenesulfonic acid monohydrate and 3 ml of absolute ethanol was stirred for 30 min. To this mixture was added a solution of 304 mg of 7-hydroxymethyl-7 / 4- benzhydryl ester. nitrobenzylidinamino / cephalosporanic acid in 3 ml ethanol and 0.5 ml methylene chloride. The mixture was stirred for 30 minutes, filtered, and after the filter cake was washed with ethanol, the filtrates were evaporated under reduced pressure at or below ambient temperature. The resulting solid was washed several times with ether and dried under a stream of nitrogen. The IR and MRI of the product correspond to the labeled structure.

D. 7-Hydroxymethyl-7-aminocephalosporanoic acid benzhydryl ester

A mixture of 3.5 ml of ether, 0.5 ml of ethyl acetate, 2 ml of water and 22 mg of dicalium hydrogen phosphate was prepared and 100 mg of 7-hydroxymethyl-7-aminocephalosporanoic acid, tosylate salt and the mixture were stirred vigorously. a few minutes. After separation of the phases, the aqueous phase is extracted with ether, the combined organic phase is dried under anhydrous magnesium sulfate and evaporated to a gummy mass under reduced pressure. The product was transferred several times by dissolving in a small amount of chloroform and again evaporating under high vacuum to a gummy mass. The resulting product shows IR and NMR spectra corresponding to the labeled structure.

E. 7- (2-Thienylacetoxymethyl) -7- (4-nitrobenzylidinamino) cephalosporic acid benzhydryl ester

A solution of 90 mg of 7-hydroxymethyl-7- (4-nitrobenzylidinamino) -cephalosporanic acid benzhydryl ester in 0.3 ml of dry methylene chloride is cooled to 0 ° C and treated with 0.5 ml of dry methylene chloride containing 100 mg of pyridine, also cooled to 0 ° C. To this was added a solution of 25 mg of 2-thienylacetyl chloride in 25 ml of dry methylene chloride was cooled and stirred for 10 minutes and kept at 0 ° C for 2 hours. The mixture was shaken with a solution of 55 mg of dicalium hydrogen phosphate in 3 ml of water, the organic phase was removed and dried with anhydrous magnesium sulfate and brought to high vacuum under high vacuum. The tire is treated by dissolving in chloroform and again evaporating to a reduced pressure tire. The product was purified by preparative thin layer chromatography on silica gel 1 mm plates with a fluorescent indicator. After developing with 5% ethyl acetate in chloroform, the desired band is outlined by shortwave and longwave UV light, separated and eluted with ethyl acetate. The product shows the IR and NMR spectra characteristic of the proposed structure.

F. 7- (2-Thienylacetoxymethyl) -7-aminocephalosporanoic acid benzhydryl ester.

A mixture of 15.9 mg of powdered 2,4-dinitrophenylhydrazine, 13.5 mg of p-toluenesulfonic acid and 2 ml of absolute ethanol was stirred for 30 min.

To this was added a solution of 58 mg of 7- (2-thienylacetoxymethyl) -7- (4-nitrobenzylidinamino) cephalosporic acid benzhydryl ester in

1.5 ml of ethanol and 0.2 ml of methylene chloride. After stirring for 30 min, the mixture was filtered and the filter cake washed with ethanol. The filtrate was evaporated under reduced pressure at or below ambient temperature and the resulting solid was washed several times with ether. The solid was shaken with a mixture of 28 mg of dicalium hydrogen phosphate, 2 ml of water and 4 ml of ether, the phases separated and the aqueous phase extracted with ether. The combined organic phases were dried with anhydrous magnesium sulfate and evaporated to a gum under high vacuum. The product was purified by preparative thin layer chromatography on 1 mm silica gel with a fluorescent indicator, using 30% ethyl acetate in benzene as eluent. After localization of the desired band using both shortwave and longwave UV light, it is scraped and eluted with ethyl acetate. The product exhibits the desired IR and NMR absorptions and is substantially homogeneous under thin layer chromatography.

G. 7hydroxymethyl-7-2-thienylacetamide / cephalosporic acid benzhydryl ester

I By O -> N acyl migration

7- (2-Thienyl-acetoxymethyl) -7-aminocephalosporanoic acid benzhydryl ester undergoes spontaneous O -> N acyl migration. When the migration progresses satisfactorily, as is understood by thin layer chromatography, the more polar product 7-hydroxymethyl-7- (2-thienylacetamido) -cephalosporanic acid benzhydryl ester can be isolated by preparative thin layer chromatography on silica. It displays all the desired features in

IR or NMR spectra and is homogeneous according to thin layer chromatography.

II By direct acylation with anhydride

A mixture of 100 g of 7-hydroxymethyl-7-aminocephalosporic acid benzhydryl ester, tosylate salt and 43 mg of 2-thienylacetic anhydride was vigorously stirred with a mixture of 2 ml of water, 3.5 ml of ether, 0.5 ml of ethyl acetate and 30 mg of dicalium hydrogen phosphate. The phases were separated and the aqueous phase was re-extracted with ether, the organic phases were combined and concentrated to 0.5 ml. After the addition of 0.1m pyridine, the reaction mixture was allowed to stand at room temperature for 18 hours and evaporated to high vacuum. The oil was mixed with 5 ml of ether and shaken with a mixture of 25 mg of dicalium hydrogen phosphate in 2 ml of water. After separation of the phases and re-extraction of the aqueous phase with 3 ml of ether, the combined organic phases were dried under anhydrous magnesium sulfate and evaporated to an oil under high vacuum. The crude product was washed twice by dissolving in a small volume of chloroform and then again evaporated under high vacuum, then purified by flash chromatography. The material obtained is identical in every respect to the regroup reaction obtained.

III By direct acylation in acid chloride

A solution of benzyl ester of 7x id roxomethyl-7-m noocephalone spore new acid, obtained from 114 mg of tosylate salt in 0.2 ml, dried on a methylene chloride sieve, was cooled to 0 ° C. To it was added dropwise 33 mg of 2-thienylacetyl chloride in 0.2 ml dried on a sieve of methylene chloride with stirring for 30 sec, then 16 mg of pyridine also added dropwise in 0.2 ml of dried on a sieve of methylene chloride. The mixture was stirred for 1 h at 0 ° C, then evaporated to a gum in a stream of dry nitrogen. The tire was shaken vigorously with a mixture of 77 mg of dicalium hydrogen phosphate, 3.5 ml of ether, 0.5 ml of ethyl acetate and 2 ml of water. After phase separation, the aqueous layer was again extracted with ether and the combined organic phases were dried with anhydrous magnesium sulfate, filtered and evaporated under reduced pressure. The resulting gum was purified by preparative thin layer chromatography on 1000 μl silica gel with a fluorescent indicator. The desired band is detected by shortwave UV light, removed and eluted with ethyl acetate. The product exhibits all the physical and spectral properties expected for the desired compound.

N. Sodium salt of 7-hydroxymethyl-7- (2-thienylacetamide) -cephalosporic acid

A mixture of 59 mg of 7-hydroxymethyl-7- (2-thienylacetamido) -cephalosporanoic acid benzhydryl ester, 0.5 ml of anisole and 1.0 ml of trifluoroacetic acid was left at room temperature for 10 min, after which the mixture was concentrated under reduced pressure to oil. The product was dissolved in 5 ml of chloroform and shaken with a mixture of 5 ml of water and 8.4 mg of sodium bicarbonate. The phases were separated and the organic phase was again washed with water. The combined aqueous phases were washed with methylene chloride and then lyophilized to give 42 mg of solid product, with IR and NMR spectral properties coinciding with the desired product.

Example 126

7-Methoxymethyl 7- (2-furylacetamide) -cephalosporanoic acid sodium salt.

A. 7-Hydroxymethyl-7- (2- (2-furyl) -acetamide) -cephalosporanoic acid benzhydryl ester

To a stirred, ice-cooled solution of 0.350 g of benzhydryl ester of 7-hydroxymethyl-7-aminocephalosporanate in 1.0 ml of dry methylene chloride was added dropwise over a period of 30 sec. 0.119 g (2-furyl) -acetyl chloride in 0.1 ml dry methylene chloride and then 0.065 g pyridine in 0.5 ml dry methylene chloride. The reaction mixture was stirred for 60 min at 0 ° C, then poured into 10 ml of methylene chloride, covered with 10 ml of ice water and stirred. The organic phase was separated, washed with dilute aqueous dicalium hydrogen phosphate, water and saturated sodium chloride solution and dried under anhydrous magnesium sulfate. Evaporation of methylene chloride under reduced pressure gave a gum which was purified by preparative thin layer chromatography on 1000 μg silica gel GF plates. The desired band is visualized by shortwave UV light, removed and eluted with ethyl acetate. Evaporation of the ethyl acetate under reduced pressure gave 7-hydroxymethyl-7- (2- (2-furyl) -acetamido / cephalosporanic acid benzhydryl ester.

B. 7-Methoxymethyl-7- (2- (2-furyl) -acetamido) cephalosporic acid benzhydryl ester. A stirred solution of 0.334 g of benzhydryl 7-hydroxymethyl-7 (2- (2-furyl) -acetamido) -cephalosporanic acid in 2 ml of methylene chloride was held at -5 ° C until 7.5 μl of boron trifluoridate was added. Then slowly, at the same temperature, a solution of diazomethane in methylene chloride was added. The reaction is vigorous, with the formation of a white solid (polymetallic) and the addition ends when a slight yellow color * persists in the solution for a short time. After 30 min at -5 ° C, the solid was filtered and the filtrate was diluted with 10 ml of methylene chloride and washed with dilute sodium bicarbonate and water and dried over anhydrous magnesium sulfate. Evaporation of the solvent under reduced pressure gave an oil which was chromatographed on silica gel using chloroform as eluent to give 7-methoxymethyl-7- (2- (2-furyl) -acetamido / cephalosporic acid benzhydryl ester.

C. 7-Methoxymethyl-7- (2-furylacetamido) -cephalosporanoic acid sodium salt

A solution of 0.295 g of 7-methoxymethyl-7 / 2- (2-furyl) -acetamido] -cephalosporic acid benzhydryl ester in 2.5 ml of anisole is treated with 7.5 ml of trifluoroacetic acid and then kept at room temperature 10 min. Excess reagents are removed under reduced pressure at room temperature. The residue was dissolved in 20 ml of methylene chloride and stirred with a solution of 0.075 g of sodium bicarbonate in 10 ml of water. After separation of the phases, the methylene chloride layer was extracted with 5 ml of water. The combined aqueous solutions were washed with 3 portions of 5 ml of methylene chloride and lyophilized to give 7-methoxymethyl-7- (2- (2-furyl) -acetamido / cephalosporanic acid sodium salt as a powder. The acetoxymethyl group of the 7-methoxymethyl-7- (2- (2-furyl) -acetamido] -cephalosporanic acid sodium salt can be converted to other 3-substituted cephalosporins. The acetoxy group may be displaced by reacting with nitrogen bases and other compounds of greater nucleophilicity than the oxygen nucleophile. In addition, 3-acetoxymethyl can be catalytically reduced in the presence of hydrogen to give 3-methylcephalosporins. Other 3-substituted compounds such as 3-hydroxymethyl can also be prepared and converted into other valuable compounds.

Example 127.

7-Carboxymethyl-7- (2-thienylacetamido) -cephalosporanic acid disodium salt.

A. 2, 2, 2-Trichloroethyl-7- (4-nitrobenzalidimine) -cephalosporic acid

0.800 g of 2, 2, 2-trichloroethyl-7-aminocephalosporanic acid was dissolved in 200 ml of C ₆ H ₆ , and 0.300 g of p-nitrobenzaldehyde was added. The mixture was heated at reflux with azeotropic removal of the formed water. After 1 h, the benzene solution was evaporated under reduced pressure to give a foam.

B. 2, 2, 2-trichloroethyl-7- (2, 2, 2 trichloroethoxycarbonylhydroxymethyl) -7- (4-nitrobenzaldimine) -cephalosporanate.

0.550 g of 2, 2, 2-trichloroethyl-7- (4-nitrobenzaldimine) -cephalosporanate was dissolved in 5 ml of dimethylformamide under nitrogen. 0.205 g of 2, 2, 2-trichloroethyl glyoxalate in 2 ml of dimethylformamide was added dropwise over a period of 15 minutes. The reaction mixture was stirred for 1/2 hour and the dimethylformamide was evaporated under reduced pressure to give the product.

C. 2, 2, 2-trichloroethyl-7- (2, 2, 2 trichloroethoxycarbonylhydroxymethyl) 7-am and nocephalosporanate

A mixture of 0.200 g of 2,4-dinitrophenylhydrazine, 0.171 g of p-toluenesulfonic acid, monohydrate and 6 ml of absolute ethanol was stirred for 30 min. A solution of 0.600 g of 2, 2, 2-trichloroethyl-7- (2, 2) was added to the mixture. , 2-tri98 chloroethoxycarbonylhydroxymethyl (7- (4-nitrobenzaldimine) -cephalosporanic acid in 6 ml ethanol and 0.5 ml CH ₂ Cl ₂ . The mixture was stirred for 30 min, filtered and after the filter cake was completely washed with ethanol, the filter and washes were evaporated under reduced pressure to give a solid product.

The solid was suspended in 15 ml of water containing 155 mg of K ₂ HPO ₄ and extracted three times with methylene chloride (20 ml). The combined organic phases were dried over magnesium sulfate and evaporated under reduced pressure to give the product gum.

D. 2, 2, 2-trichloroethyl-7- (2, 2, 2 trichloroethoxycarbonylhydroxymethyl) -7- (2-thienylacetamido) cephalosporic acid

0.620 g of 2, 2, 2-trichloroethyl-7- (2, 2, 2-trichloroethoxycarbonylhydroxymethyl) -7-aminocephalosporanoic acid was dissolved in 100 ml of CH ₂ Cl ₂ , cooled to 0 ° C and treated with 0.160 g 2- thienylacetyl chloride, then 0.100 g of pyridine in 2 ml of CH ₂ Cl ₂ was added dropwise. The mixture was stirred for 1 h at 0 ° C and poured onto crushed ice. After stirring, the organic phase was separated and washed once with phosphate buffer pH 2 and once with water. After drying over magnesium sulfate, the solvent is removed to give a product which is purified by chromatography on silica gel.

E. 2, 2, 2-trichloroethyl-7- (2, 2, 2 trichloroethoxycarbonylmethylsulfonyloxymethyl) -7- (2-thienylacetamide) -cephalosporanate

0.350 g of 2, 2, 2-trichloroethyl-7- (2, 2, 2-trichloroethoxycarbonylhydroxymethyl) -7- (2-thienylacetamido) -cephalosporanate was dissolved in 10 ml of CH ₂ Cl ₂ and cooled to 0 ° C, added.

0.115 g of methanesulfonyl chloride and then 0.080 g of pyridine. The reaction mixture was stirred at 0 ° C for 1 1/2 hours. The reaction mixture was poured onto crushed ice and stirred. The organic phase was separated and washed with phosphate buffer with pH 2 and water, dried and evaporated. The crude product was purified by chromatography on silica gel.

F. 2, 2, 2-trichloroethyl-7- (2, 2, 2 trichloroethoxycarbonyl methyl methyl) -7 (2-thienylacetamidocephalosporanate)

0.300 g of 2, 2, 2-trichloroethyl-7- (2, 2, 2-trichloroethoxycarbonylmethylsulfony loxy methyl) -7- (2-thienylacetamido) -cephalosporanate were dissolved in 10 ml of DMBO. 0.100 g of sodium iodide was added and the mixture was stirred at room temperature overnight. Dimethyl sulfoxide is removed under reduced pressure. The residue was partitioned between 10 ml of water and 10 ml of CH ₂ Cl ₂ . The organic phase was washed once with water, dried and evaporated to give a crude product which was purified by preparative thin layer chromatography on silica gel plates.

G. 2, 2, 2-trichloroethyl-7- (2, 2, 2 trichloroethoxycarbonylmethyl) -7- (2-thiophenacetamide) -cephalosporanate

0,200 g of 2, 2, 2-trichloroethyl-7- (2, 2, 2-trichloroethoxycarbonylidmethyl) 7- (2-thiophenacetamido) -cephalosporanate are dissolved in 20 ml of ethanol, 0.100 g of sodium acetate and 0.100 g of 5% are added. palladium on calcium carbonate and the mixture was treated with hydrogen at 40.6,865 kN / m ^{2 for} 2 h. The catalyst was filtered off. The filtrate was evaporated under reduced pressure and the residue was chromatographed on silica gel to give the product.

H. 7-Carbomethoxy-7- (2-thienylacetamido) -cephalosporanic acid disodium salt. 0,300 g 2, 2,

2-Trichloroethyl-7- (2, 2, 2-trichloroethoxycarbonyl methyl) -7- (2-thiophenacetamido) -cephalosporanoic acid was dissolved in 15 ml of 95% acetic acid. 0.300 g of zinc powder are added in 5 equal portions over a period of 20 min and the mixture is stirred vigorously at room temperature. The reaction mixture was carefully poured onto ice-cold phosphate buffer with pH 2 and extracted with ethyl acetate. The ethyl acetate extract was washed once with water, dried and evaporated. The residue was mixed with 10 ml of methyl isobutyl ketone and extracted with 15 ml of water containing 0.120 g of NaHCO ₃ . The organic phase was separated and washed once with water. The combined aqueous phases were washed twice with methylene chloride and dried by cooling to give the crude product. Crystallization from MeOH / isopropanol gives the pure product.

Example 128

A. 7-Azido7-trifluoromethylcephalosporanoic acid benzhydryl ester

A degassed solution of 7-diazocephalosporanic acid benzhydryl ester 45 mg and 144 mg CF ₃ I in 15 ml of benzene, saturated with triethylammonium azide, was irradiated with UV light> 300 mm in a pyrex flask until a diazorose at 4.8 μ in IR spectrum. The resulting solution was washed with water, dried over magnesium sulfate, evaporated and carefully chromatographed on silica gel eluting with chloroform, yielding both epidimers of 7-diazo-7-trifluoromethyl-cephalosporic acid benzhydryl ester.

B. 7β-8ΜΗ-7a-trifluoromethylcephalosporanate benzhydryl ester

7β- (2-Thienylacetamide) 7-a-trifluoromethylcephalosporanoic acid benzhydryl

7β-83ΗΛθ7a-Trifluoromethyl cephalosporanic acid benzhydryl ester 542 mg is hydrogenated at

40.6,895 kN / m ² for 3 h using 1 g of platinum oxide in 50 ml of dioxane containing 500 mg of thienylacetic anhydride. ϊβ-οβεΛΠΗεΗΗετο is formed during hydrogenation and acylation in situ. After filtration of the catalyst, the solution was treated for 15 minutes with 0.5 ml of water to hydrolyze the excess anhydride, followed by vacuum distillation. The crude product was mixed with 25 ml of ethyl acetate, washed with aqueous bicarbonate, dried with magnesium sulfate, filtered, evaporated and chromatographed on silica gel using 4: 1 chloroform-ethyl acetate eluent. 7β- (2-Thienylacetamide) -7a-trifluoromethylcephalosporanic acid benzhydryl ester was obtained. The other epimer produces the epimeric compound by the same procedure.

C. 7β- (2-Thienylacetamide) -7a-trifluoromethylcephalosporanoic acid sodium salt

7β- (2-phenyl-acetamide) -7a-trifluoromethylcephalosporanoic acid benzhydryl ester 640 mg was dissolved in 0.8 g of anisole and cooled to 0 ° C. 4 ml of trifluoroacetic acid cooled to 0 ° C was added. allowed to continue for 2 min at 0 ° C. A 0.1 mm vacuum was immediately applied and the reaction was allowed to warm to room temperature without further heating. The anisole was then removed by distillation at 30 ° C / 0.1 mm. A few milliliters of anisole is added to the residue and withdrawn at 30 ° C / 0.1 mm to ensure complete removal of the trifluoroacetic acid. The product was treated with 5 ml of water containing 84 mg of sodium bicarbonate and se

100 lyophilized. The resulting powder was washed thoroughly with ether and dried to give the desired sodium salt. The other epimer gives the epimeric sodium salt by the same procedure.

Example 129

7a-Difluoromethyl-7- (2-thienylacetamido) -cephalosporanic acid sodium salt

A. 7β- (ρnitrobenzylidenamine) 7α-difluoromethylcephalosporanic acid benzhydryl ester

7- (p-Nitrobenzylidenamine) -cephalosporanic acid benzhydryl ester 571 mg was dissolved in 20 ml

1,2-dimethoxyethane (DME) containing 1 ml of chlorodifluoromethane. With vigorous stirring under nitrogen, 112 mg of potassium butoxide was slowly introduced as a suspension into DME for about 1 hour.

After stirring for another hour, the KCI was filtered off and the low-boiling fractions were removed in vacuo to give 7β- / ρ-nitrobenzylidenamine / 7-α-difluoromethyl-cephalosporanic acid benzhydryl ester. It can be purified, if desired, by chromatography on silica gel, eluting with 2% ethyl acetate in chloroform.

B. 7a-Difluoromethyl-7- (2-thienylacetamido) -cephalosporanic acid sodium salt. Removal of the active Schiff base with aniline hydrochloride, acylation of the 7-aminocephalosporan compound obtained with thienylacetyl chloride or other acid chloride or anhydride, and deblocking of the 4-carboxy group with trifluoroacetic acid, all carried out as obtained 7adifluoromethyl-7- (2-thienylacetamido) -cephalosporanoic acid sodium salt

Example 130

7- (2-thienylacetamido) 7-aminocephalosporanoic acid.

Step A. 7,7-Diazodecephalosporanic acid benzhydryl ester

7-Bromo-7-azidocephalosporanic acid benzhydryl ester (110 mmol) was dissolved in 0.5 M lithium azide 20 ml in dimethylformamide and heated at 68 ° C for 3 min under nitrogen. The solution was diluted with water 100 ml and extracted with chloroform 20 ml. The chloroform layer was washed with two 60 ml portions of water and dried over sodium sulfate. Evaporation of the solvent in vacuo gave crude 7,7-diazidocephalosporanic acid benzhydryl ester.

Step B. 7amino-7-azidocephalosporanic acid benzhydryl ester

7,7-Diazidocephalosporanic acid benzhydryl ester 10 mmol was dissolved in ethyl acetate and 3 g of 10% palladium on carbon was added. The mixture was hydrogenated at atmospheric pressure at 45 ° C for 1 hour. A crude isomeric mixture of 7-amino-7-azidocephalosporanic acid benzhydryl ester is obtained.

Step C. 7/2-Thienylacetamido / -7-azidocephalosporanoic acid benzhydryl ester

An isomeric mixture of 7-amino-7-azidocephalosporanic acid benzhydryl ester 5 mmol obtained in step B is dissolved in methylene chloride 50 ml and cooled to 0 ° C, 2 thienylacetyl chloride 5 mm is added and the solution is left at 0 ° C for 1 hour . The methylene chloride solution was washed with saturated sodium bicarbonate solution, 2 portions of 50 ml of 1 N hydrochloric acid, 1% sodium bicarbonate solution of 50 ml and water of 50 ml. The solvent was dried over sodium sulfate and evaporated to give 7-2-thienylacetamido / -7-azidocephalosporanoic benzhydryl as gum.

Step D. 01101/2-Thienylacetamido / -7-aminocephalosporanoic acid benzhydryl ester

7- (2-Thienylacetamido) -7-azidocephalosporanoic acid benzyl ester 100 mmol obtained in step C was dissolved in ethyl acetate 50 ml and 3 g of palladium-on-carbon catalyst were added. The hydrogenation is carried out at room temperature at

40.6,895 kN / m ² 1 hour. The catalyst was removed by filtration and the solvent was evaporated in vacuo to give N- (2-thienylacetamide) -7-aminocephalosporanic acid benzhydryl ester.

Step E. 7- (2-Thienylacetamido) -7-aminocephalosporanoic acid

7- (2-Thienylacetamido) -7-aminocephalosporanoic acid benzhydryl ester was dissolved in anisole 10 ml at 0 ° C. To the solution was added trifluoroacetic acid 15 ml, cooled to 0 ° C, and the mixture was aged at room temperature for 1 h. The excess trifluoroacetic acid and anisole were removed by evaporation and the residue was washed twice with chloroform and evaporated to dryness. The crude product obtained is 7- (2-thienylacetamido / 7-aminocephalosporanoic acid).

Example 131

7- (2-thienylacetamido) -7-azidocephalosporanoic acid

7- (2-Thienylacetamido) -7-azidocephalosporanoic acid 1 mm was dissolved in anisole 10 ml at 0 ° C. To this solution was added trifluoroacetic acid 15 ml, cooled to 0 ° C, and the mixture was allowed to age at room temperature for 1 h. The excess trifluoroacetic acid and anisole were removed by evaporation and the residue was washed twice with chloroform and evaporated to dryness. The resulting crude product and 7- (2-thienylacetamide) -7-azidocephalosporanic acid.

Example 132

7- (2-thienylacetamidocephalosporanoic acid)

7- (2-Thienylacetamido) -7-aminocephalosporanic acid 10 ml was dissolved in methanol 50 ml at room temperature. 3.5 dimethylamidinpyrazole 10 ml was added and the mixture was stirred at room temperature for 16 h. The solution was diluted with 100 ml of water and extracted three times with ether. The ether layer was discarded and the aqueous layer was lyophilized to give crude 7- (2-thienylacetamido) -7-guanidinocephalosporanic acid.

Example 133

7- (2-thienylacetamido) -7-ureidocephalosporanic acid

7- (2-Thienylacetamido) -7-aminocephalosporanic acid 10 mmol was dissolved in dry pyridine 20 ml and the solution was cooled to -10 ° C. Carbamoyl chloride 10 mm dissolved in 20 ml of dry pyridine was slowly added and the mixture was stirred for 1 hour at -10 ° C, then quenched with ice-cold saturated sodium bicarbonate solution 100 ml. The aqueous solution was extracted with ether to remove pyridine and the aqueous layer was lyophilized. The salt precipitate was suspended with methanol and filtered and the methanolic solution evaporated to dryness to give crude 7- (2-thienylacetamido) -7-ureidocephalosporanic acid.

Example 134

7- (2-thienylacetamido) -7-N, N-dimethylureido / cephalosporic acid

Similarly, N, N-dimethylcarbamoyl chloride is used instead of carbamoyl chloride to give 7- (2-thienylacetamido) -7- (N, N-dimethylureido) -cephalosporanic acid. Replacing the carbamoyl chloride of Example 133 with an equivalent amount of 102 V, N-dimethylcarbamoyl chloride according to the procedures described in this example yields 7- (2-thienylacetamido / 7-N, N-dimethylureido) -cephalosporic acid.

Example 135

7- (2-thienylacetamido) -7- (N-amidinoureido) -cephalosporic acid

7- (2-Thienylacetamido) -7-aminocephalosporanoic acid 10 ml was dissolved in a solution of dimethylformamide 50 ml and pyridine 20 ml and cooled to 0 ° C. The solution was treated with 100 ml of toluene solution containing 17% phosgene (large excess). The reaction mixture was allowed to stand for 2 h and evaporated in vacuo to remove toluene and excess phosgene. The resulting solution containing 7- (2-thienylacetamide) -7- (chloroformamido) cephalosporic acid was then cooled to 0 ° C and guanidine 10 ml was added. The solution was aged at 0 ° C for 2 h and quenched by the addition of saturated sodium bicarbonate solution. The mixture was extracted with ether to remove the specimen and water was removed by evaporation in vacuo and the salt precipitate was extracted with dry methanol. Evaporation of methanol gave crude 7- (2-thienylacetamido-7 / Mamidinureido / cephalosporic acid).

Example 136

7- (2-thienylacetamido) -7-sulfonamidocephalosporanoic acid

7- (2-Thienylacetamido) -7-aminocephalosporanic acid 10 ml was dissolved in pyridine 50 ml and cooled to 0 ° C. To this solution was added pyridine sulfur trioxide complex 10 mmol. The mixture was allowed to stand at 0 ° C for 3 hours and the pyridine was removed in vacuo to give crude 7- (2-thienylacetamido) -7-sulfo-namidocephalosporic acid.

Example 137

7- (2-thienylacetamido) -7-acetamidocephalosporanoic acid 7- (2-thienylacetamido) -7-aminocephalosporanoic acid 1 mmol was dissolved in 10 ml of water at 0 ° C containing one millimol of sodium bicarbonate at 0 ° C. The mixture was treated with acetic anhydride 2 mmol, stirred at 0 ° C for 1 h, and acidified with dilute hydrochloric acid 1 mmol. The precipitated product was filtered off, washed with water and dried in vacuo to give crude 7- (2-thienylacetamido) -7-acetamidocephalosporanic acid.

Example 138

7- (2-thienylacetamido) -7-methanesulfonamidocephalosporic acid

Substitution of an equivalent amount of methanesulfonyl chloride with acetic anhydride of Example 137 and the procedure described therein gave 7- (2-thienylacetamido) -7-methanesulfonamidocephalosporic acid.

Example 139

7- (2-thienylacetamido) -7- (ethoxycarbonylamino) -cephalosporic acid

Step A. 7-Azido-7- (ethoxycarbonylamino) cephalosporic acid benzhydryl ester

Ethylurethane 25.0 g was melted at 48 ° C and then 7-bromo-7-azidocephalosporanic acid 5 mm benzhydryl ester was added. One portion of ethylurethane 15 g containing silver tetrafluoroborate 15 mm was also added to the solution. The temperature was maintained at 50 ° C for 5 min and quenched by the addition of a large volume of ether. The resulting silver bromide was removed by filtration through a 103 fusor soil and the ether solution was washed successively with water 400 ml, saturated sodium bicarbonate solution 400 ml and two 400 ml portions of water. The organic layer was dried with sodium sulfate, the solvent was evaporated in vacuo and the gum residue triturated several times with water to remove excess ethylurethane. 7-Azido-7- (ethoxycarbonylamino) cephalosporic acid benzhydryl ester is thus obtained.

Step B. 7- (2-Thienylacetamido) -7- (ethoxycarbonylamino) cephalosporanic acid benzhydryl ester

2-thienylacetic anhydride 5 ml, pyridine 0.1 ml and 10% palladium on carbon 3.0 g were added to a solution of 7-azido-7- (ethoxycarbonylamino / cephalosporic acid benzhydryl ester 5 mm dissolved in ethyl acetate 25 ml. The mixture was hydrogenated at room temperature for 1 h. The catalyst was then removed by filtration and the residue was evaporated in vacuo to a glassy mass. Thus, crude 7- (2-thienylacetamido) -7-ethoxycarbonylamino / cephalosporanic acid benzhydryl ester is obtained.

Step C. 7- (2-Thienylacetamido7- (ethoxycarbonylamino) cephalosporic acid

7- (2-Thienylacetamido) -7- (ethoxycarbonylamino) cephalosporanic acid benzhydryl ester 1.0 mm, obtained in step B, was dissolved in 10 ml of anisole at 0 ° C. To this solution was added trifluoroacetic acid 15 ml, cooled to 0 ° C and the mixture was left at room temperature for 1h. The excess trifluoroacetic acid - anisole was removed by evaporation and the residue was treated twice with chloroform and evaporated to dryness. The crude product thus obtained is 7 (2-thienylacetamido) -7- (ethoxycarbonylamino) cephalosporic acid.

Example 140 7- (2-Thienylacetamido) -7-methylaminocephalosporanoic acid

Step A. 7-Azido-7-nitrocephalosporanic acid benzhydryl ester

7-Bromo-7-azidocephalosporanic acid benzhydryl ester 10 mmol was heated at 68 ° C for 4 min in dimethylformamide 60 ml containing lithium nitrite 10 mm. The solution was diluted with water 300 ml and extracted with 50 ml portions of chloroform. The chloroform layer was washed with three milliliter portions of water and dried over anhydrous sodium sulfate. The crude product obtained is 7-azido-7-nitrocephalosporanic acid benzhydryl ester.

Step B. 7- (2-Thienylacetamido) -7-nitrocephalosporanic acid benzhydryl ester

7-Azido-7-nitrocephalosporanic acid benzhydryl ester 10 ml was dissolved in isopropanol 25 ml at 0 ° C and sodium borohydride 5 mm dissolved in isopropanol 25 ml was added to this solution. The solution was left at 0 ° C for 1 hour and then poured into ice water. The solution was extracted with three 50 ml portions of methylene chloride and the organic layer was dried over sodium sulfate. To the dried methylene chloride solution was added ten millimoles of 2-thienylacetic anhydride 10 mm and colidine 10 mm at 0 ° C. After standing at 0 ° C for 30 min, the methylene chloride layer was extracted with two 50 ml portions of 0.1 N hydrochloric acid and two 50 ml portions of sodium bicarbonate and covered with two 50 ml portions of water. The methylene chloride solution was then dried over sodium sulfate and evaporated in vacuo to give a gummy product. The crude product obtained is 7- (2-thienylacetamido) -7-nitrocephalosporanic acid benzhydryl ester.

Step C. 7- (2-Thienylacetamido) 7-nitrocephalosporanic acid

After substitution of an equivalent amount of 7- (2-thienylacetamido) -7-nitrocephalosporanoic acid 7- (2-thienylacetamido) -7- (ethoxycarbonylamino) -cephalosporanic acid benzhydryl ester in Example 139, step C, and following the procedure, given there, 7- (2-thienylacetamido) -7-nitrocephalosporanic acid is obtained.

Step D. 7- (2-Thienylacetamido) 7-aminocephalosporanoic acid

7- (2-Thienylacetamido) -7-nitrocephalosporanic acid 10 mm is reduced in methanol 50 ml using a palladium-on-carbon catalyst 1.0 g at 40.6,895 kN / m ² at room temperature for 1 h. The catalyst is removed by filtering. The solvent was then removed by evaporation to give the crude 7- (2-thienylacetamido) -7-aminocephalosporanic acid.

Step E. 7- (2-Thienylacetamido / 7-methylaminocephalosporanoic acid)

7- (2-Thienylacetamido) -7-aminocephalosporanic acid 100 mm To this solution was added a platinum oxide catalyst of 0.1 g and formaldehyde of 5 mm. Hydrogenation is carried out at room temperature under hydrogen pressure for two atmospheres for 5 to 10 minutes. The catalyst is filtered off and the solvent is removed 10 in vacuo to give crude 7/2-thienylacetamido / -7-methylaminocephalosporanic acid.

Example 141

Following the procedure described in Example 140, step A to E, all 7-amido-7 / alkylamino / cephalosporanate products of the present invention can be obtained. For example, by substituting an equivalent amount of aralkane anhydride or heterocyclic anhydride with the 2-thienylacetic anhydride mentioned in step B and after substituting an equivalent amount of alkaldehyde or araldehyde with formaldehyde mentioned in step 7, the syntheses thereof can be synthesized in step E. (N-alkylamino- and N-aralkylamino / cephalosporic acids). The following accompanying table equations illustrate this process and the products obtained from it:

sn.osn,

UNEP, 4 @]

NaBH. (RC0), 0 ¹¹ V ° * s η

RCNH-l - «'' S'S || p.c-COOH o ^ -j ^^ sn, ----- ►

COOCH, -f ^]

II NO _:

R-CNH-I— b Ns ^ J-CH.OCCH,

COOH

II RCNH

_n RCHO II --► RCNH

CH.OCCH,

O II

CH.OCCH,

COOH

105

TABLE 1

R CH ₂ R ' nooo-ch4 ^ 2 ^ 4 dream ₃ -with ₂ n ₅ O- ^CH = - / dream, /, dream, O ~ ^cH . О-сн ₂ -CH / CH _3/2 0- ™, - - SI / -O About ^_s · -0-a 0-dream ₂ - / CH ₂ / _g CH ₃ О ~ ^сн > -0-dream,

Example 142

7- (2-thienylacetamido) -7- (N, dimethylamino) -cephalosporanoic acid

7- (2-Thienylacetamido) -7-aminocephalosporanic acid 10 mmol was dissolved in acetic acid 100 ml containing formaldehyde 40 mm. Platinum oxide catalyst 0.2 g was added to this solution and hydrogenation was carried out over 4-6 atmospheres of hydrogen at 35 ° C for 1 h. The catalyst was then filtered and the solvent was evaporated in vacuo. Crude 7- (2-thienylacetamido 7- (N, N-dimethylamino) -cephalosporanic acid was obtained.

Example 143

7-Phenylacetamido-7- (ethoxycarbonylamino) -cephalosporic acid

Step A. 7-Phenylacetamido-7- (ethoxycarbonylamino) -cephalosporanic acid benzhydryl ester

Phenylacetic anhydride 5 mmol, pyridine 0.1 ml and Bolhoffer catalyst 3.0 g were added to a solution of 7-azido-7 (ethoxycarbonylamino) -cephalospor4545 acid 5 mm dissolved in ethyl acetate 25 ml. The mixture was hydrogenated at room temperature for 1 hour. The catalysts were then removed by filtration and the residue was evaporated in vacuo. 7-Phenylacetamido7- (ethoxycarbonylamino) cephalosporic acid crude benzhydryl ester is obtained.

Step B. 7-Phenylacetamido-7 (ethoxycarbonylamino) -cephalosporic acid

7-Phenylacetamido-7- (ethoxycarbonylamino) cephalosporanic acid benzhydryl setter 1.0 ml was dissolved in anisole 10 ml at 0 ° C and to this solution was added trifluoroacetic acid 15 ml cooled to 0 ° C. The mixture was aged at room temperature for 1 h. The excess trifluoroacetic acid - anisole was removed by evaporation to dryness to give crude 7-phenylacetamido

7- (ethoxycarbonylamino) -cephalosporanic acid.

Example 144

Follow the procedure in the example

142, for the preparation of 7- (2-thienylacetamido) -7- (N, N-dimethylamino) -cephalosporanic acid, all 7-amido-7- (N, N-dialkylamino) -cephalosporanic acids can be obtained according to the invention. For example, by substituting an equivalent amount of alkaldehyde instead of the formaldehyde cited in Example 142 and using the procedure described therein, the following 7- (2-thienylacetamido) -7- (N, dialkylamino) -cephalosporanic acids can be obtained. The following equation and Table 2 illustrate the process and the resulting products:

R ²

-with ₂ n ₅

- / CH _2/2 CH ₃

TABLE 2

OCHR ² osn ₃ osn ₂ sn ₃ OSN-

107

OCH-CH - / A

OCH / CH _2/2 CH ₃ OCH / CH _2/3 CH ₃

-CH-CH - / CH, / ₃ CH,

- / CHj / .CH,

Example 145

7-Ethoxycarbo-10 nylamino-7- (2-thienylacetamido) -cephalosporanoic acid sodium salt

Step A. 7-Azido-7- (ethoxycarbonylamino) -cephalosporanic acid benzhydryl ester 15

7-Azido-7-bromocephalosporanic acid benzhydryl ester of 3.9 g was added to 36 g of ethylcarbamate while maintaining the temperature at 65 ° C. To the resulting mixture was added 20 portions of molten silver fluoroborate 3.6 g dissolved in ethylcarbamate 18 g and the reaction mixture was maintained at 67-70 ° C for 5 min. The mixture was then poured into ether with 25 stirring and the resulting suspension is filtered through celite to remove silver bromide. The ether was extracted successively with water 100 ml, saturated aqueous sodium bicarbonate solution 100 ml and two portions of water 100 ml. The extracted ether solution was dried over sodium sulfate and evaporated under reduced pressure. The resulting residue was then treated three times with a small amount of water and then dissolved in chloroform. The chloroform solution was dried over sodium sulfate and evaporated to dryness. 2.1 g of 7-azido-7- (ethoxycarbonylamino) -cephalosporanic acid benzhydryl 40 ester is obtained.

Step B. 7amino-7- (ethoxycarbonylamino) -cephalosporanic acid benzhydryl ester. 45

7-Azido (ethoxycarbonylamino) -cephalosporic acid benzhydryl ester 1.0 g was dissolved in dioxane 100ml. Platinum oxide 1.0 g was added and the reaction mixture was stirred under hydrogen at atmospheric pressure for 1 h. Another 1.0 g platinum oxide was added and the reaction mixture was again placed under hydrogen and stirred for 3 h until the azide reacted, establishes by aliquot analysis of aliquots. The solvent was removed under reduced pressure and the residue was mixed with chloroform 50 ml and filtered through silica gel G into chloroform in a 60 ml sintered glass funnel. The resulting product was eluted in chloroform to collect 200 ml of chloroform. The chloroform was then removed under reduced pressure to give 7-amino-7- (ethoxycarbonylamino) cephalosporic acid benzhydryl ester 0.6 g.

Step C. 7ethoxycarbonyl and lami-7- (2-thienylacetamido) -cephalosporanoic acid benzhydryl ester

7-Amino-7 (ethoxycarbonylamino) -cephalosporanic acid benzhydryl ester 0.6 g obtained in step B is mixed with methylene chloride 25 ml and cooled to 0 ° C. 2-thienylacetyl chloride is added dropwise over 30 sec. 0.6 ml, 0.038 mol and 60 sec later, rilidine 0.6 ml, 0.01 mol was added. The reaction mixture was stirred at 0 ° C for 15 min and poured onto crushed ice. The mixture was stirred and the organic layer was separated and washed once with water 20 ml, once with sodium bicarbonate 5% 20ml and

108 once with water 20 ml. The methylene chloride mixture was then dried and evaporated to dryness to give 1.42 g of crude 7-ethoxycarbonylamino-7- (2-thienylacetamido) -cephalosporic acid benzhydryl ester. The product was transferred to a column of 60 g of silica gel under benzene and the column was eluted with benzene, 100 ml fractions were collected and then 300 ml of methylene chloride - benzene 1: 1 in three fractions and 500 ml of methylene chloride in five fractions. The product exits the column eluting with chloroform 400 ml in four fractions. 7-Ethoxycarbonylamino-7- (2-thienylacetamido) -cephalosporanic acid benzhydryl ester is obtained. This product was mixed with methylene chloride 25 ml and stirred at room temperature with 20 ml of sodium bicarbonate solution of 0.120 g in water for 0.5 h. The resulting layers were separated and the organic layer was washed with water, dried and evaporated to dryness. . 0.4 g of 7ethoxycarbonylamine-7- (2-thienylacetamido / cephalosporanic acid benzhydryl ester) are obtained.

Step D. 7-Ethoxycarbonylamino-7- (2-thienylacetamido) -cephalosporanic acid sodium salt

7-Ethoxycarbonylamino-7- (2-thienylacetamido) -cephalosporanic acid benzhydryl ester 0.4 g was dissolved in anisole 2.5 ml and treated with trifluoroacetic acid 10 ml at room temperature for 10 minutes. The trifluoroacetic acid was removed and the anisole removed. under reduced pressure, keeping the temperature below 40 ° C, the residue was stirred with chloroform 25 ml and treated with 20 ml of water containing 0.120g of sodium bicarbonate. The mixture was stirred for 0.5 h at room temperature and the organic phase was separated and washed with water. The combined aqueous phases were then washed with methylene chloride and lyophilized to give 0.32 g of the 7-ethoxycarbonylamino-7- (2-thienylacetamido) -cephalosporanoic acid sodium salt as a brownish solid.

Example 146

7-Ethoxycarbonylamino-7- (2-thienylacetamido) -3-pyridinedecephalosporanoic acid

7-Ethoxycarbonylamino-7- (2-thienylacetamido) -cephalosporanic acid sodium salt was dissolved in 3 cc. buffer solution obtained by dissolving potassium iodide 30 g in water 20 ml containing 5 ml pyridine and adjusting the pH to 6.5 with dilute acetic acid. The resulting reaction mixture was stirred at 80 ° C for 2 h and then cooled to room temperature. The solution was then freeze-dried, dissolved in 25% acetic acid 40 ml, filtered and the filtered solution electrophoresed. 7-Ethoxycarbonylamino7- (2-thienylacetamido) -3-pyridinedecephalosporanic acid is obtained.

Example 147

7-Ethoxycarbonylamino-7- / phenylacetamido-cephalosporic acid sodium salt

By reacting benzhydryl 7-amino-7-ethoxycarbonyl-aminocephalosporanic acid with phenylacetyl chloride instead tienilatsetilhlorid as described in Example 145 above, gave benzhydryl 7-ethoxycarbonylamino-7-fenilatsetamidotsefalosporanova acid. Using the procedures described in Example 145, this ester is converted to the sodium salt.

Example 148

7-Ethoxycarbonylamino-7-thiophenoxyacetamidocephalosporanic acid sodium salt

109

By reacting benzhydryl 7-amino-7-etoksikarbonilaminotsefalosporanova acid with an equivalent amount feniltioatsetilhlrorid instead tienilatsetilhlorid as described in Example 145, to give benzhydryl 7etoksikarbonilamino-7-tiofenoksiatsetamido-cephalosporanic acid. Using the procedure described in Example 145 above, this ester can be converted to the sodium salt.

Example 149

7-Ethoxycarbonylamino-7- (2-furylacetamido-cephalosporanoic acid) sodium salt

7-Ethoxycarbonylamino-7- (2-furylacetamido) -cephalosporanic acid sodium salt is obtained by reacting 7-amino-7-ethoxycarbonylaminocephalosporanic acid with furylacetyl chloride and then converting this ester into a sodium salt, as described 145.

Example 150

7-Amino-7- (2-cyanilacetamido) -cephalosporanic acid sodium salt

Reaction of 7-azido-7-bromocephalosporanic acid benzhydryl ester with tert-butylcarbamate instead of ethylcarbamate, as described in Example 145, gave 7-tert-butylcarbonylamino-7-aminocephalosporanate ester. Treatment of this product, as described in Example 145, yields the 7-amino-7- (2-thienylacetamido) -cephalosporanic acid sodium salt.

Example 151

7-Hydrazino-7- (2-thienylacetamido) -cephalosporanic acid sodium salt

Exposure to 7-azido-7-bromocephalosporanic acid with tert-butylcarbamate instead of ethylcarbamate as described in Example 145 and the resulting product reacted as described in Example 145 yielded the 7-hydrazino-7- / 2- sodium salt thienylacetamido / cephalosporanic acid.

Example 152

7-Azido-7- (2-thienylacetamido) -cephalosporanic acid sodium salt

Step A. 7-Azido-7- (2-thienylacetamido / cephalosporanic acid benzhydryl ester)

7-Azido-7-bromocephalosporanic acid benzhydryl ester 3.6 g was added to 2-thienylacetamide 30 g and the mixture was heated to 65-70 ° C. To this reaction mixture was added silver trifluoroborate 3.6 g and 2-thienylacetamido 15 g and the mixture was heated to 65-70 ° C for 10 min. The reaction mixture was poured onto ether 200 ml with stirring and the resulting suspension filtered through diatomaceous earth to remove silver bromide. The ether solution was extracted with water 100 ml, saturated aqueous sodium bicarbonate solution 100 ml and two 100 ml portions of water. The ether layer was then dried over sodium sulfate and evaporated under reduced pressure. The resulting residue was treated twice with small amounts of water and then dissolved in chloroform 25 ml. After drying the chloroform solution over sodium sulfate, it was evaporated under reduced pressure to give 4 g of 7-azido-7- (2-thienylacetamido) -cephalosporic acid benzhydryl ester.

Step B. 7-Azido 7- (2-thienylacetamido) -cephalosporanic acid sodium salt

The 7-azido-7/2-thienylacetamido) -cephalosporanic acid benzhydryl ester obtained in step A was blocked by reaction with trifluoroacetic acid and anisole and the product was separated as the sodium salt according to the procedure described in Example 145.

Example 153

When the method of Example 130, step A, is carried out by proceeding from 7-chloro-7-azidocephalosporanic acid benzhydryl ester, using an equivalent amount of potassium or sodium azide instead of lithium azide, 7,7-diazocephalosporalosporin acid benzhydryl ester is obtained. . Reduction of this product, acylation of the 7-amino compound with phenylacetyl chloride and cleavage of the benzhydryl group by the procedure described in the above examples give 7- (2-furylacetamido-7-aminocephalosporic acid).

Example 154

7-Bromo-7-phenylacetamidocephalosporanic acid sodium salt

Step A. The 7-aminocephalosporanic acid benzhydryl ester

P-toluenesulfonic acid monohydrate 4.3 g, 0.22 mol was added with stirring and room temperature to a suspension of 7-aminocephalosporanic acid 6.8 g, 0.25 mol, in peroxide-free dioxane (300 ml) . The clear solution was concentrated in vacuo and washed twice with dioxane. The residue was dissolved in dioxane 300 ml at room temperature and a solution of phenyldiazomethane 10 g, 0.05 mol, in dioxane 25 ml was added dropwise over a period of 15 minutes. The resulting wine-colored solution was stirred for a further 30 min and methanol was added 25 ml to destroy excess diphenyldiazomethane. The mixture was concentrated in vacuo and the residue partitioned between methylene chloride 200 ml and water 200 ml containing dicalcium phosphate 10 g, pH 8.5. The organic phase was washed with water, dried over sodium sulfate and concentrated in vacuo to give an oil. The oil was stirred with ether 100 ml for one hour and the resulting precipitate was filtered off, washed with ether and dried to constant weight, 4.7 g, 43% yield. The compound obtained is 7-aminocephalosporanic acid benzhydryl ester, mp 126128 ° C.

Analysis: Calculated: C, 63.0; H, 5.01; N, 6.37

Found: C, 62.7; H, 5.18; N, 5.18.

IR to chloroform: 5.6 μ / β-lactam C = 0 / and 5.8 μ / ester C = 0 /.

NMR in deuterated chloroform: 1.85 tau (singlet), ΝΗ ₂ ; 2.0 tau (singlet), / CH ₃ ~ C = 0 /; 3.45 tau (doublet), (CH ₂ S); 4.8 tau (singlet), (CH ₂ OAc); 4.7 tau (doublet), / C ₆ H; 4.9 tau (doublet), (C ₇ H); 6.98 tau / doublet /, / θ ^ ΟΗ; and 7.4 tau (singlet), (phenyl).

Step B. Benzhydryl ester of 7diazocephalosporanic acid

To a mixture of sodium nitrite 1.6 g, water 30 ml and methylene chloride 40 ml was added 7-aminocephalosporanic acid benzhydryl ester 880 mg, 0.002 mol, with stirring at 0 ° C. A solution of ptoluenesulfonic acid 760 mg, 0.004 mol, in water 5 ml was then added over a period of several minutes and the mixture was stirred at 0 ° C for 20 min. The organic phase was then separated and washed with ice water for 10 cc, dried. over sodium sulfate at 0 ° C, filtered and concentrated in vacuo at room temperature. 900 mg of 7-diazocephalosporanic acid benzhydryl ester is obtained as a glass. The IR is 4.8 μ / strong Ν = Ν /, 5.6 μ / β lactam C = 0 /, and 5.8 μ / ester C = 0 /.

NMR in deuterated chloroform: 2.0 tau (singlet) / CH ₃ -C = 0 /; 3.4 tau

111 (doublet), (CH ₃ S); 4.8 tau (singlet), (CH ₂ OAc); 5.6 Tau (singlet), (C ₆ H); 6.98 tau (singlet), (CH) and 7.4 tau (singlet) (phenyl).

Step C. 7bromo-7-azidocephalosporanoic acid benzhydryl ester

Preparation of triethylammonium azide. To the suspension of sodium azide

1.5 g in water 5 ml and methylene chloride 10 ml maintained at -10 ° C was added 4 ml 50% sulfuric acid dropwise over a period of 5 min. The mixture was stirred for another 5 min and the organic phase was poured out of the water paste and the aqueous extract was washed with methylene chloride 5 cc. The combined organic phase was dried over potassium chloride and the decanted azide solution was adjusted to pH 7 with tertilamine. The resulting triethylammonium azide was stored at -10 ° C.

Preparation of bromazide. To methylene chloride 8 ml, maintained at 0 ° C, was added sodium azide 2.66 g, 0.04 mol, and then bromine 9.65 g, 0.0042 mol. To this mixture was added dropwise concentrated hydrochloric acid 12 mol with stirring at 0 ° C, stirring continued for another 3 h at 0 ° C. The organic layer was decanted and the aqueous layer was extracted with methylene chloride 5 ml. The extracted bromazide was stored at -10 ° C.

To a solution of 7-diazocephalosporanic acid benzhydryl ester 900 mg in methylene chloride 20 ml and nitromethane 10 ml maintained at 010 ° C was added a solution of triethylammonium azide as obtained above and then a bromazide solution was obtained as obtained above. Then 50 ml of water was added followed by a sufficient amount of solid sodium bicarbonate to reach the pH of solution 8. The organic phase was separated and extracted with two 20 ml portions of water, dried over sodium sulfate and concentrated under vacuum to give 900 mg, 83%, 7-bromo-7-azidocephalosporanic acid benzhydryl ester.

Thin layer chromatography on silica gel with chloroform showed a major stain for this product at R _f 0.2. Chromatography of 900 mg of the product on 25 gms of silica gel with chloroform gave 400 mg, 39%, of a one-spot product as an oil.

IR to chloroform: 4.7 μ / Ν ₃ /, 5.56 μ / β-lactam C = 0 / and 5.75 μ / ester C = 0 /.

NMR in deuterated chloroform: 2.0 tau (singlet), (CH _{3 -} C = 0); 3.38 tau (singlet), (CH ₂ S); 4.7 tau (singlet), (CH ₂ O); 4.9 (singlet) (C ₆ H); 6.9 tau (singlet), (CH) and 7.4 tau (singlet), (phenyl).

Step D. 7bromo-phenylacetamidocephalosporanic acid benzhydryl ester

Sodium borohydride 0.1 g was added to a solution of 7-azido-7-bromocephalosporanic acid benzhydryl ester 0.5 g in bis- (2-methoxyethyl) -ether 10 ml and the resulting mixture was stirred overnight under nitrogen. The mixture was then poured onto ice water, the pH adjusted to 8, extracted with methylene chloride, dried and evaporated. 7-Amino-7-bromocephalosporanic acid benzhydryl ester is obtained.

The 7-amino-7-bromocephalosporanic acid benzhydryl ester is then treated with two equivalents of phenylacetic acid anhydride in methylene chloride solution and the resulting mixture is stirred for half an hour and then poured into ice-cold water. The resulting aqueous solution was extracted with methylene chloride, the solvent was dried over sodium sulfate and evaporated under reduced pressure to give 7-bromo-7-phenylacetamidcephalosporanic acid benzhydryl. This product was further purified by chromatography on silica gel. 7-Bromo-7-phenylacetamidocephalosporanoic acid benzhydryl ester was obtained.

Step E. 7-Bromo-7-phenylacetamidocephalosporanoic acid sodium salt

7-Bromo-7-phenylacetamidocephalosporanic acid benzhydryl ester 0.4 g was dissolved in anisole 3.5 ml and treated with trifluoroacetic acid 10 ml at room temperature for 10 minutes. Trifluoroacetic acid and the anisole were removed under reduced pressure while maintaining the temperature under reduced pressure. 40 ° C. The residue was stirred in chloroform 25 ml and treated with water 20 ml containing 0.120 g sodium bicarbonate. The resulting mixture was stirred for half an hour at room temperature and the organic phase was separated and washed with water. The combined aqueous phase was washed twice with methylene chloride and lyophilized to give the 7-bromo-7-7-phenylacetamidocephalosporanic acid sodium salt as a brownish solid.

Example 155

7-Chloro-7- (2-thienylacetamido / cephalosporic acid) sodium salt

Step A. 7-Azido-7-chlorocephalosporanic acid benzhydryl ester

To a solution of 7-diazocephalosporanic acid benzhydryl ester 900 mg in methylene chloride 20 ml and nitromethane 10 ml at 0-10 ° C was added triethylammonium azide prepared as described in Example 154, step C, followed by the addition of the chloride wall, prepared as described below. To this reaction mixture was added 50 ml of water and enough solid sodium bicarbonate to bring the pH of the mixture to 8. The organic layer was separated and extracted twice with 20 ml portions of water, dried over sodium sulfate and concentrated under reduced pressure. 7-Azido-7-chlorocephalosporanic acid benzhydryl ester is obtained.

Preparation of chlorazide

A solution of 5.25% 100 ml of sodium hypochloride in water was mixed with chlorine 50 ml and cooled to -10 ° C. Sodium azide 4.3 g was added to this mixture, followed by the addition of aqueous acetic acid solution 10 ml, prepared by diluting 4 ml glacial acetic acid to 10 ml. Acetic acid was added over a period of 15 min and the reaction mixture was stirred for an additional 15 min at -10 ° C. The aqueous phase was then separated, frozen by cooling in a solid carbon dioxide acetone bath and the organic phase decanted and dried over anhydrous sodium sulfate. This methylene chloride solution is then used in the reaction described above.

Step B. 7-Chloro-7- (2-thienylacetamido / cephalosporic acid) sodium salt

7-Azido-7-chlorocephalosporanic acid benzhydryl ester obtained in step A is reduced with sodium borohydride as in Example 154, step D to give 7-amino-7-chlorocephalosporanic acid benzhydryl, and this product is then reacted by acylation with 2-thienylacetic acid anhydride. 7chloro-7- (2-thienylacetamido / cephalosporanoic acid benzhydryl ester was obtained using 7-chloro-7113/2-thienylacetamido / cephalosporic acid benzhydryl ester instead of 7-bromo-7-phenylacetate 154 , step E, and following the procedure described therein, produces the 7-chloro-7- (2-thienylacetamido) -cephalosporanic acid sodium salt.

Example 156

7-Fluoro-7- (2-furylacetamido / cephalosporic acid) sodium salt

Step A. 7-Azido-7-fluorocephalosporanic acid benzhydryl ester

To a solution of 7-azido-7-bromocephalosporanic acid benzhydryl ester of 0.5 g in anhydrous acetonitrile 50 ml was added tetraethylammonium fluoride 3.0 g. The resulting mixture was stirred overnight at room temperature and then poured into water and extracted with methylene chloride to give 7-azido-7-fluorocephalosporanic acid benzhydryl ester.

Step B. 7-Fluoro 7- (2-furylacetamido / cephalosporanic acid) sodium salt

7-Azido-7fluorocephalosporanic acid benzhydryl ester according to step A, is reduced with sodium borohydride, 7-amino-7-fluorocephalosporanic acid benzhydryl ester is obtained and this product is then acylated by reaction with the anhydrate obtained with anhydrite. 7-Fluoro-7- (2-furylacetamido / cephalosporanic acid benzhydryl ester was converted to the sodium salt following the procedure described in Example 154, step E.

Example 157

7- (2-Thienylacetamido) -7-chlorocephalosporanic acid sodium salt

Stage A. Benzhydryl ester of

7-Aminocephalosporanic acid

7-Aminocephalosporanic acid 272 mg was suspended for 5 min at 250 ° C in dioxane 7 ml with p-toluenesulfonic acid monohydrate 170 mg. Methanol 2 ml was added, the solvents were removed in vacuo and dioxane was added twice and evaporated in vacuo. Dioxane 8 ml was then added followed by the addition of diphenyldiazomethane 290 mg. After the nitrogen evolution is complete, the solvent is distilled off in vacuo and the residue is stirred with methylene chloride 10 ml and water 10 ml containing sufficient dibasic potassium phosphate to bring the mixture pH to

8. The layers were separated and the aqueous portion was extracted twice with methylene chloride. The combined organic layers were dried with sodium sulfate, filtered and then evaporated to give an oily crystalline material. Washing with ester gave 150 mg, 35%, yielding 7-aminocephalosporanic acid benzhydryl ester as a dry solid, melting point 110-115 ° C.

Step B. 7- (p-Nitrobenzylidenamine / cephalosporanic acid benzhydryl ester)

7-Aminocephalosporanic acid benzhydryl ester 438 mg was refluxed for one hour in benzene 50 ml with p-nitrobenzaldehyde 151 mg in an azeotropic drying apparatus. The solvent was then eliminated by vacuum distillation to give 571 mg of the crude benzhydryl ester of 7- (p-nitrobenzylidenamino) cephalosporic acid. This compound can be used directly in the next step or, if desired, can be purified by recrystallization

114 from a mixture of one part benzene and two parts cyclohexane.

Step C. 7- (p-Nitrobenzylidenamine) -7-chlorocephalosporanic acid benzhydryl ester. 7- (p-Nitrobenzylidenamine / cephalosporic acid benzhydryl ester) 571 mg was stirred at 0 ° C under nitrogen in dimethylformamide 5 ml. A catalytic amount of potassium tert-butoxide was added and then tert-butyl hypochloride 109 mg was added under rapid stirring for one to two minutes.

The solvent was evaporated in vacuo and the resulting oily residue was then stirred in benzene 25 ml, washed four times with 10 ml water, dried with magnesium sulfate and then filtered and evaporated. 7- (p-Nitrobenzylidenamino) -7-chlorocephalosporanic acid benzhydryl ester was obtained. The crude material thus obtained can be used directly in the next step, or optionally purified by chromatography on neutral silica gel, eluting with 2% ethyl acetate in chloroform.

Step D. 7-Amino-7-chlorocephalosporanic acid benzhydryl ester, hydrochloride

7- (p-Nitrobenzylidenamino) -7-chlorocephalosporanic acid benzhydryl ester 606 mg and aniline hydrochloride 260 mg were stirred together for one hour at 25 ° C in methanol 10 ml. The methanol was removed at 0.1 mm pressure and 30 ° C and then the residue was coated with ether and allowed to stand for 1 h to crystallize. The solid was triturated with ether, filtered and then washed with ether. 7-amino-7-chlorocephalosporanic acid hydrochloride benzhydryl ester is obtained. This material contains some unreacted aniline hydrochloride, but is pure enough to be used directly, i.e. without further purification in the next step.

Step E. 7- (2-Thienylacetamido) -7-chlorocephalosporanoic acid benzhydryl ester

7-Amino-7chlorocephalosporanic acid benzhydryl ester, the hydrochloride obtained in step E, was added to methylene chloride 25 ml and this mixture was stirred vigorously at -10 ° C. Triethylamine 0.5 g was added slowly to give the free amine, i.e., amino-7-chlorocephalosporic acid benzhydryl ester. Then 2-thienylacetyl chloride 0.5 g was added and the reaction mixture was allowed to warm to room temperature. The excess acid chloride is then hydrolyzed by shaking with water and the methylene chloride layer is washed sequentially with a diluted phosphoric acid solution, water and dilute sodium bicarbonate solution, to pH 2. After drying with magnesium sulphate, the solution is filtered and evaporated to give a mixture of 7- (2-thienylacetamido) -7-chlorocephalosporanoic acid benzhydryl ester and N-phenyl-2-thienylacetamido. 7- (2-Thienylacetamido) -7-chlorocephalosporanic acid pure benzhydryl ester was obtained by chromatography on silica gel, eluting said mixture with 4 parts chloroform and 1 part ethyl acetate.

Stage F. 7- (2-Thienylacetamido) -7-chlorocephalosporanic acid sodium salt

7- (2-Thienylacetamido) -7-chlorocephalosporanic acid benzhydryl ester was dissolved in anisole 0.8 g and cooled to 0 ° C. Pre-cooled to 0 ° C three 115 fluoroacetic acid 4 ml was added and the reaction was allowed to proceed at 0 ° C for 2 min. Vacuum 0.1 mm mercury is applied immediately and the reaction mixture is allowed to warm to room temperature without external heating. The anisole is then removed at 30 ° C and 0.1 mm mercury and to the residue is added several ml of anisole which is removed at 30 ° C and 0.1 mm mercury to ensure complete elimination of the trifluoroacetic acid. The product was treated with water 5 ml containing 84 mg of sodium bicarbonate and then lyophilized. The resulting powder was washed thoroughly with ether and dried. 7- (2-Thienylacetamido) -7-chlorocephalosporanic acid pure salt was obtained.

Example 158

7- (2-Thienylacetamido-7-bromocephalosporanoic acid) sodium salt

Step A. 7- (2-Thienylacetamido) -7-bromocephalosporanic acid benzhydryl ester

7-Bromo-7-azidocephalosporanoic acid 543 mg to 2-thienylacetic anhydride 500 mg and platinum oxide catalyst 1.0 g in dioxane 50 ml were added and the mixture was hydrogenated at atmospheric pressure for 4 h. was filtered to remove the platinum oxide catalyst, water was added 0.5 ml and the solution was left to stand for 15 min to hydrolyze all unreacted acetic anhydride. The solution was then distilled in vacuo and the crude 7- (2-thienylacetamido) -7-bromocephalosporanoic acid crude benzhydryl ester was dissolved in ether 25 ml, washed with aqueous sodium bicarbonate, dried over magnesium sulfate, filtered and evaporated. The resultant product is 7- (2-thienylacetamido) -7-bromocephalosporanic acid benzhydryl ester and can optionally be further purified by chromatography on silica gel eluting with four parts chloroform and one part ethyl acetate.

Step C. 7- (2-Thienylacetamido) -7-bromocephalosporanic acid sodium salt

7- (2-Thienylacetamido) -7-bromocephalosporanic acid benzhydryl ester was dissolved in anisole of 0.8 mol and cooled to 0 ° C. Pre-cooled to 0 ° C trifluoroacetic acid 4 ml was added and the mixture was allowed to react at 0 ° C for 2 min. The reaction mixture was then immediately placed under vacuum and allowed to warm to room temperature without external heating. The mixture was then vacuum distilled, with the anisole distilled at 30 ° C / 0.1 mm mercury. The resulting product was treated with 5 ml water containing 84 mg of sodium bicarbonate and lyophilized to a powder, gently washed with ether and dried. 7- (2-Thienylacetamodi) -7-bromocephalosporanic acid sodium salt was obtained.

Example 159

7- (2-Thienylacetamido) -7-chlorocephalosporanic acid sodium salt

Step A. 7- (2-Thienylacetamido) -7-chlorocephalosporanic acid benzhydryl ester

Substitution of 7-chloro-7-azidocephalosporanic acid benzhydryl ester 499 mg with the 7-bromo-7-chlorocephalosporanic benzhydryl ester mentioned in Example 158, and following the procedure described therein, yields the 7-benzhydryl ester (2-Thienylacetamido) -7-chlorocephalosporanova

116 acid.

Step C. 7- (2-Thienylacetamido) -7-chlorocephalosporanoic acid sodium salt

By substituting 7- (2-thienylacetamido / 7-chlorocephalosporanoic acid benzhydryl ester) with 7- (2-thienylacetamido) -7-bromocephalosporanoic benzhydryl ester of Example 158, step B, and following the procedure described therein, gives the sodium salt of 7- (2-thienylacetamido-7-chlorocephalosporanic acid).

Example 160

7- (α- (α-Aminophenyl) acetamido) -7- (dimethylphosphono) cephalosporic acid sodium salt

Step A. 7-Azido-7-dimethylphosphono / cephalosporic acid benzhydryl ester

To a solution of 7-azido-7-bromocephalosporanic acid benzhydryl ester 72 mg, 0.0005 mmol, in dimethoxymethane 10 ml was added silver dimethyl phosphate 120 mg, 0.0005 mmol, prepared by mixing equimolar amounts of dimethyl phosphite and silver and filtering the slow-forming solid. The mixture was stirred overnight at room temperature, protected from light, air and moisture. The inorganic material is filtered and the filter is concentrated at low temperature and pressure. The residue was dissolved in methylene chloride and the solution was washed with cold dilute aqueous sodium bicarbonate. It is dried and concentrated under reduced pressure. The residue was redissolved in a small amount of chloroform and the mixture was purified by chromatography on silica gel using chloroform as eluent. The original eluate contains little of the starting material. The second eluting fraction was concentrated under reduced pressure. 7-Azido-7- (diethylphosphono) cephalosporic acid benzhydryl ester was obtained.

Step B. 7-amino-7- (dimethylphosphono) cephalosporic acid benzhydryl ester

7-Azido-7 / dimethylphosphono / cephalosporanic acid benzhydryl ester 200 mg was dissolved in dioxane 20 ml and hydrogenated at room temperature and normal pressure for 1 h using platinum oxide 200 mg as catalyst. A fresh portion of 200 mg of catalyst was added and the hydrogenation continued for an additional 2 h. The solvent was removed at low temperature and pressure. The dry residue was dissolved in chloroform and the solution filtered by suction through a pad of silica gel to remove the suspended catalyst used. The silica gel was rinsed with chloroform carefully to elute any adsorbed material. The filtrate was concentrated under reduced pressure to give 7-amino-7- (dimethylphosphono / cephalosporic acid benzhydryl ester, suitable for use in the next step without further purification.

Step C. 7-α- (α-Aminophenyl) -acetamido-7- (dimethylphosphono) cephalosporic acid benzhydryl ester

A solution of 7-amino-7- (dimethylphosphono / cephalosporic acid benzhydryl ester) 273 mg in methylene chloride 15 ml was stirred at 0 ° C and α-amino-α-phenylacetyl chloride hydrochloride 220 mg was added. Pyridine 0.5 ml was then added immediately and the mixture stirred at 0 ° C for 15 min and then poured onto crushed ice containing a slight excess

117 of hydrochloric acid. The layers were separated and the organic phase was washed with an excess of cold dilute aqueous sodium bicarbonate solution. It was dried and concentrated to give 7β- (α-aminophenyl) acetamido-7- (dimethylphosphono) cephalosporic acid benzhydryl ether. The crude material was purified by chromatography on silica gel with chloroform as an elution agent. A small amount of the 7-α- (α-aminophenyl) -acetamido-7- (dimethylphosphono) cephalosporanic acid isomer isomer is also isolated from the chromatographic mixture.

Step D. 7-β- / α-Aminophenyl / acetamido-7- / dimethylphosphono / cephalosporanic acid sodium salt

A solution of 7-β- (α-aminophenyl / acetamido-7- (dimethylphosphono) cephalosporic acid benzhydryl ester 70 mg and anisole 11 mg in methylene chloride 0.5 ml was stirred in an ice bath. Add in increasing portions over a period of five minutes a cold solution of trifluoroacetic acid 23 mg in methylene chloride 0.2 ml. After another 30 min, the mixture was concentrated by a high vacuum pump. The residue was taken up in benzene 5 ml and 3 ml and aqueous sodium bicarbonate solution 8.4 ml was added. The mixture was then stirred well at room temperature for 15 min, the layers were separated and the organic phase was stirred once more with water 2 ml, then the layers were separated again. The combined aqueous extracts were washed once with ether and lyophilized. 7-p- (α-Aminophenyl) acetamido-7 (dimethylphosphono) cephalosporic acid is obtained.

Example 161

7 - // Dimethylamino / methoxyphosphinyl // - 7- (α-2-furylacetamido) -cephalosporanic acid sodium salt

Stage A. Benzhydryl ester of

7-Azido-7 - // dimethylamino (methoxyphosphinyl) -cephalosporic acid

A mixture of 7-azido-7-bromocephalosporanic acid benzhydryl ester 545 mg, N, N, O-trimethylphosphonamidic acid 246 mg and silver tetrafluoroborate 195 mg in dimethoxyethane 5 ml was stirred overnight at room temperature, protected from air. The solution was filtered from inorganic salts and concentrated to dryness under reduced temperature. The residue was dissolved in methylene chloride and the solution was washed with cold aqueous sodium bicarbonate solution, then with water and dried and concentrated. The residue was redissolved in chloroform and the solution was applied to a column of silica gel suspended in chloroform. Elution of the column with chloroform and concentration of the eluate afforded the 7-azido-7 - [(dimethylamino) -methoxyphosphinyl] -cephalosporanic acid benzhydryl ester.

Step B. 7amino-7 - // dimethylamino (-methoxyphosphinyl) -cephalosporanic acid benzhydryl ester

7-Azido-7 // dimethylamino / methoxyphosphinyl / cephalosporanic acid benzhydryl ester 250 mg in dioxane 25 ml was hydrogenated at room temperature and at normal pressure using a 500 mg platinum oxide catalyst, which was added in two equivalent portions in The hydrogenation is continued for 2-3 hours after the second addition or until the azide disappears, analyzed by IR spectroscopy. The solvent is completely removed under reduced temperature and pressure. The residue was dissolved in chloroform, the spent catalyst was removed by filtration through a layer of silica gel, and the adsorbent was washed thoroughly with chloroform. The filtrate was washed under reduced pressure to give 7-amino-7 - [dimethylamino (methoxyphosphinyl) -cephalosporanic acid benzhydryl ester.

Step C. 7 // Dimethylamino / -methoxyphosphinyl / 7-a- (2-furylacetamido) cephalosporanic acid benzhydryl ester

7 - // Dimethylamino (methoxyphosphinyl) -cephalosporanic acid benzhydryl ester solution 280 mg in methylene chloride 15 ml was stirred at 0 ° C and 2-furylacetyl chloride 140 mg was added. Pyridine 0.5 ml was then added immediately and stirring at 0 ° C continued for 15 min. The reaction mixture was poured onto crushed ice containing excess hydrochloric acid. The layers were separated and the organic phase was washed with an excess of cold dilute aqueous sodium bicarbonate solution, then washed once with water, dried and concentrated. The crude product was purified by chromatography on silica gel with chloroform as eluent. Concentration of the eluent afforded 7-dimethylamino-methoxyphosphinyl / 7-p- (2-furylacetamido) -cephalosporanoic acid benzhydryl ester together with a small amount of 7 - // dimethylamino / -methoxyphosphinyl) -7-a- (2-furylacetamido / cephalosporanic acid).

Step D. 7 - // Dimethylamino (-methoxyphosphinyl) -7-β (2-furylacetamido) -cephalosporanic acid sodium salt

A solution of 7 - // dimethylamino (methoxyphosphinyl) -7-β- (2-furylacetamido) -cephalosporic acid benzhydryl ester 65 mg and anisole 11 mg in methylene chloride 0.5 ml was stirred at 0 ° C. A cold solution of 23 mg trifluoroacetic acid 23 mg in methylene chloride 0.2 ml was added 3-4 minutes. After a further 30 min at 0 ° C, the mixture was concentrated to dryness under severely reduced pressure, the residue was mixed with benzene 5 ml and 3 ml of aqueous sodium bicarbonate solution 8.4 mg was added. The mixture was stirred well at room temperature for about 15 min and the layers were separated. The organic phase was extracted once with water and the combined aqueous phases were washed once with ether and lyophilized to give 7 - [dimethylamino] -methoxyphosphinyl] -7H- (2-furylacetamido) cephalosporic acid.

Example 162

7f- (α-Carboxyphenyl) -acetamido-7- (bis) dimethylamino (phosphinyl) -cephalosporanic acid disodium salt

Step A. Azido-7- (bis) -dimethylamino (-phosphinyl) -cephalosporanic acid benzhydryl ester

A mixture of 7azido-7-bromocephalosporanic acid benzhydryl ester 545 mg, tetramethyldiamidophosphoric acid 273 mg and silver tetrafluoroborate 195 mg in dimethoxyethane 5 ml was stirred overnight at room temperature, protected from light, air and moisture. The reaction mixture was filtered and the filtrate was concentrated to dryness under reduced pressure. The residue was dissolved in methylene chloride and the solution was washed with cold dilute sodium bicarbonate solution and then dried and concentrated. The residue was dissolved in a little chloroform and applied to a silica gel column from which the product was eluted with chloroform. The eluate was concentrated under reduced pressure to give 7-azido-7- (bis) -dimethylamino (-phosphinyl) -cephalosporanoic acid benzhydryl ester.

Step B. Amino-7- (bis (dimethylamino) -phosphinyl) -cephalosporanic acid benzhydryl ester

7-Azido-7 (bis (dimethylamino) -phosphinyl) -cephalosporanic acid benzhydryl ester 250 ml was dissolved in dioxane 25 ml and hydrogenated at room temperature and normal pressure for 3/4 h using a platinum oxide catalyst 250 mg. Fresh catalyst 250 mg was added and the hydrogenation continued for 2-3 h or until the azide disappeared, which was established with an IR spectrophotometer. The mixture was concentrated to dryness under reduced pressure and the residue was mixed with chloroform and filtered by suction through a pad of silica gel to remove the spent catalyst. The silica gel was then washed thoroughly with chloroform and the filtrate was concentrated under reduced temperature and pressure. 7-Amino-7- (bis (dimethylamino) -phosphinyl) -cephalosporanic acid benzhydryl ester is obtained.

Step C. Ίβ-α-tert-Butoxycarbonylphenyl / acetamido-7- (bis) dimethylamino / phosphinyl) -cephalosporanic acid benzhydryl ester

7-Amino-7 (bis (dimethylamino) -phosphinyl) -cephalosporanic acid benzhydryl ester obtained in step B was dissolved in methylene chloride 20 ml and α- (tert-butoxycarbonyl) -phenylacetyl chloride was added at 010 mg to 0 ° C at 0 ° C. solution. The addition is followed by the immediate addition of pyridine 1 ml. After 15 min, the reaction was stopped by pouring on crushed ice and the organic phase of the mixture was washed sequentially with cold dilute acid salt, cold dilute sodium bicarbonate solution and then with water. The mixture was dried and concentrated under reduced pressure to give 7-β-α-tert.-butoxycarbonylphenyl (-acetamido-7- (bis) dimethylamino) -phosphinyl) -cephalosporanic acid benzhydryl ester. The product was purified by chromatography on silica gel and the purified product was eluted with chloroform. This chromatographic purification also gave the 7-a (a-tert-butoxycarbonylphenyl) -acetamido-7 (bis (dimethylamino) -phosphinyl) -cephalosporanic acid isomer.

Step D. 7-β- (α-Carboxyphenyl) -acetamido-7- (bis (dimethylamino) -phosphinyl) -cephalosporanic acid disodium salt

A mixture of 7β- (α-t-butylbenzylphenyl) -acetamido-7- (bis) dimethylamino) -phosphinyl) -cephalosporanic acid benzhydryl ester 158 mg, anisole 22 mg and methylene chloride was stirred at 0 ml. A cold solution of trifluoroacetic acid 46 mg in methylene chloride 0.5 ml was added over a period of 5 min and concentrated to dryness under reduced pressure. The residue was stirred with a mixture of benzene 5 ml and ether 5 ml and 10 ml of aqueous sodium bicarbonate solution 33.6 mg was added. After stirring at room temperature for 10 min, the layers were separated and the organic phase was again stirred with water for 5 ml for another 5 min. The combined aqueous extracts were washed once with 2 ml ether and lyophilized to give 7β- (α-carboxyphenyl). -acetamido-7- (bis (dimethylamino) -phosphinyl) -cephalosporic acid.

Example 163

7-Phosphono-7β- / 2-ΤΗεΗΗΑ3Η6Τ3Μ ^ Ιθ / Η6φ3ΑΟΟΠθρ3nic acid trisodium salt

120

Step A. The 7-azido-7- (di-tert-butylphosphono) -cephalosporic acid ester free ester. A mixture of 7-azido-7-bomocephalosporanic acid benzhydryl ester 545 mg, ditret.butyl phosphite 388 mg and silver tetrafluoroborate 195 mg in dimethoxyethane 5 ml was stirred overnight at room temperature, protected from air, light and moisture. The mixture was then filtered through insoluble material and concentrated to dryness under reduced temperature and pressure. The residue was redissolved in methylene chloride and the solution was washed with some ice water and then dried and concentrated. The residue was dissolved in chloroform and the solution was chromatographed on acid-washed alumina using a mixture of chloroform and ethanol as eluent. The eluate was concentrated under reduced pressure. 7-Azido-7- (diethyl-butylphosphono) -cephalosporanoic acid benzhydryl ester was obtained.

Step B. 7amino-7-di-tert-butylphosphono / -cephalosporanic acid benzhydryl ester. 7-Azido-7- (di-tert-butylphosphono) -cephalosporanic acid benzhydryl ester 110 mg was dissolved in dioxane 10 ml and hydrogenated at room temperature and normal pressure for 1 h using a platinum oxide catalyst 100 mg. A second portion of 100 mg catalyst was added and the hydrogenation continued for 2-3 h. The solvent was completely removed under reduced pressure and the residue redissolved in chloroform. The spent catalyst was removed by filtration through a pad of silica gel and the filter cake was washed thoroughly with chloroform. The filtrate was concentrated under reduced pressure to give the benzhydryl ester of

7-amino- (di-tert-butylphosphono) -cephalosporanic acid, which is ready for use without further purification.

Eta C. Diethyl tert-butylphosphono-7-N- (2-thienylacetamido) -cephalosporanic acid benzhydryl ester. 7-Amino7- (di-tert-butylphosphono) -cephalosporanic acid benzhydryl ester obtained in step B was dissolved in methylene chloride 10 ml and the solution was stirred at 0 ° C. To this solution was added 2-thienyloxyacetic acid 32 mg dicyclohexyl dicyclohexyl dicyclohexyl dicyclohexyl dicyclohexyl dicycloxy 41 mg. The reaction mixture was then allowed to warm slowly to room temperature and stirred overnight. The dicyclohexylurea which formed is removed by filtration and the filtrate is concentrated under reduced pressure. The residue was dissolved in chloroform and the solution was applied to a column filled with acid-washed alumina. 7-Di-tert-butylphosphono-7-P (2-thienylacetamido) -cephalosporanic acid benzhydryl ester was obtained by elution with a dilute methanol solution in chloroform. The product was obtained by concentration of the eluate. A small amount of the 7-di-tert-butylphosphono-7-a- (2-thienylacetamido) -cephalosporanic acid isomer is also obtained.

Step D. 7-Phosphono-7- (α-2-thienylacetamido) -cephalosporanic acid trisodium salt. A mixture of 7-di-tert-butylphosphono-7-p- (2-thienylacetamido / cephalosporanoic acid benzhydryl ester 70 mg, anisole 33 mg and methylene chloride 1 ml was stirred at 0 ° C and a cold trifluoroacetic acid solution was added 70 mg in methylene chloride 0.2 ml. After about 30 minutes, the solution was concentrated to dryness under reduced pressure, and the residue was suspended in a mixture of benzene 3 ml and ether 3 ml was 5 ml aqueous sodium bicarbonate solution 50 mg was added. The mixture was stirred well and the layers separated. The organic phase was washed with 5 ml of water and the combined aqueous solutions were washed with ether and lyophilized. 7-Phosphono-7-p- (2-thienylacetamido) -cephalosporanic acid trisodium salt was obtained.

Example 164

By the procedure described in Example 163, steps A-D, all 7-amido-7-phosphonocephalosporanic acids according to the invention can be obtained. For example, substitution with an equivalent amount of suitable phosphite and acyl halide of di-tert-butyl phosphite and 2-thienylacetic acid, cited in steps A and C of Example 163, and carrying out the procedure as described in this example, yields the corresponding salts of Gamido-7-phosphonocephalosporic acid. The following reaction scheme of the accompanying Table IV illustrates this method of preparation:

SNC, + P (OR ')

dream ₂ pers

COOH (C, H,) ₂

COOH (C »H,),

RCOH

CH ₂ OSSN ₂ ---- ►

COOH (C, H ₅ ) ₂

R-CNH

II ^{0 about}

COOH (CH,) ₂

CFjCOOHe ^.

dream, ossn, - ¹ ---- ►

R-C-NH II

Ο l

som

CH ₂ OSSN ₃

TABLE 4

R R ⁵ X m base U -CH, - 2 -with ₂ n ₅ Br On NaOH nose-dream- - / dream, /, dream, CI K. knso ₃ ^Онс < -Sn ₃ Br On NaHCO ₃ noos-ss / s ₆ n ₅ / - ^C 2 ^H 5 CI Li LiOH noos-ss / s _b n ₅ / - - / dream, /, dream, I On At ₂ CO ₃

Example 165

7- (p-cyanoethyl) 7- (2-furylacetamido) -cephalosporanic acid sodium salt

A solution of 500 mg 7- (p-nitrobenzylidenamino) -cephalosporanic acid benzhydryl ester prepared as in Example 43, in a mixture of 2.5

122 ml of tert-butyl alcohol and 2.5 ml of acrylonitrile were treated with 20 ml of N, N-diisopropylethylamine with stirring under nitrogen. The green solution was initially turned to yellow-orange after stirring for 10 min. It was then concentrated under reduced pressure to 620 mg of a yellow gummy residue. This residue was chromatographed on a mixture of 48 g of active powdered silica gel and 40 g of powdered diatomaceous earth. The elution was carried out with 2 L of ether in benzene. The desired non-crystalline cyanethyl adduct is eluted after passing the second liter of eluent through the column. The product is identified as 7- (p-nitrobenzylidenamino) -7-fluoro-ethyl-cephalosporic acid benzhydryl ester. using the procedure described in the above examples. The product was converted to the 7- (p-cyanethyl) -7- (2-furylacetamido) -cephalosporanic acid sodium salt.

Example 166

7-Methyl-7- (2-thienylacetamido) cephalosporic acid sodium salt

7- (p-Nitrobenzylidenamino) -cephalosporanic acid benzhydryl ester 571 mg was dissolved in 10 ml of dry acetonitrile containing 0.950 ml of dimethyl sulfate. With vigorous stirring under nitrogen, a solution of diisopropylethylamine in 12 ml of acetonitrile was added over a period of 5 hours. The reaction mixture was stirred overnight and the solvent removed in vacuo. The residue was mixed with 50 ml of benzene and washed successively with water, 1M aqueous phosphate buffer with pH 2 and bicarbonate. After drying, filtration and removal of the solvent, the residue is a crude product, suitable for further reaction without further purification. It can be purified if desired by chromatography on silica gel eluting with 25: 1 chloroform-ethyl acetate.

It is identified as 7- (p-Netrobenzylidenamino) -7-methylcephalosporanic acid benzhydryl ester.

This product is converted to the title compound by the procedures described in the previous examples.

Example 167

7-Acetyl-7- (2-thienylacetamide) -cephalosporanic acid sodium salt

7- (p-Nitrobenzylidenamino) -cephalosporanic acid benzhydryl ester 571 mg was dissolved in 10 ml of dry acetonitrile containing 0.950 ml of acetyl chloride. With vigorous stirring under nitrogen for a period of 5 h, a solution of diisopropylenylamine in 12 ml of acetonitrile was added. The reaction mixture was stirred overnight and the solvent removed in vacuo. The residue was mixed with 50 ml of benzene and washed sequentially with water, 1M aqueous phosphate buffer with pH 2, water and bicarbonate. After drying, filtration and removal of the solvent, the residue is a crude product, suitable for further reaction without further purification. If desired, it can be purified by chromatography on silica gel eluting with 25: 1 chloroformethyl acetate. It is identified as 7- (p-nitrobenzylidenamino) -7-acetylcephalosporanic acid benzhydryl ester.

This product is converted to the title compound by the procedures described in the above examples.

Example 168

3-carbamoyloxymethyl-7-methoxy-p-phenylacetamido-3-cephem-4-carboxylic acid

Step A. 7β- (O-5-Trichloroethoxycarbonylamino-5-carboxyalkylamido) -3-carbamoyloxymethyl-7-methoxy123

3-cephem-4-carboxylic acid

The 7β- [N-5-3Μπ-5-carboxyvaleramido / -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid monosodium salt 20.5 g was dissolved in a mixture of acetone 80 ml and aqueous 10% dicalium hydrogen phosphate 240 ml. Trichloroethoxycarbonyl chloride 25 g 118 mmol in acetone 80 ml was added dropwise to this solution. During the addition, the pH of the solution was maintained at 9.1 by the gradient addition of 2.5 N sodium hydroxide solution. After 30 min, the mixture was extracted with ethyl acetate, the ethyl acetate layer was discarded, and the aqueous layer was acidified to pH 2.5 with concentrated hydrochloric acid. The precipitated product was extracted into ethyl acetate. Drying over sodium sulfate and removing the solvent in vacuo afforded the title compound as an oil.

UV: / CH ₃ OH / λ max. 262.5 ε = 5450.

NMR / solvent DMSO, d _6/5 = 3.43 / O-CH ₃ singlet / 4.73 / 2 H ₂ CC1 ₃ partly exposed / 4.81 / _CH2 ~ O, singlet / 5.12 (6-H, singlet), 4.74 (10-H ₂ , partially visible).

Step B. 7β- / D-5'-Ethoxycarbonylamino-5-carboxyvaleramido / -3-carbamoyloxymethyl-7-methoxy-3-cephem-4 carboxylic acid di-benzhydryl ester

To a solution of the above 7β- (S-5-trichloroethoxycarbonylamino-5-carboxyvaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid in ethyl acetate 500 ml was added diphenyldiazomethane 17 g in 200 ml ether. After stirring the mixture overnight, it was extracted sequentially with solutions of sodium bicarbonate and sodium chloride. The solvent was evaporated from the dry solution to give a crude product which was purified by chromatography on silica gel. A mixture of 2: 1 chloroform and ethyl acetate was used as eluant. This product shows one TLC spot.

UV: / CH ₃ OH / λ max. 2650 gm is 7000.

NMR / solvent - CDCI _3/5 = 3.45 / O-CH ₃ singlet / 3,35 / 2 H ₂ hour £ C1 ₃ partially visible / 4.69 / _CH2 ~ O, singlet / , 5.03 (6-H, singlet), -4.88 (10-H ₂ , partially visible).

Step C. 7β- [(O-5 * -tre Chloroethoxycarbonyl) and but-5'-carboxyvaleryl] -phenylacetylamino] -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic ester acid

Mixture of 7β- / D-5'-ethoxycarbonylamino-5-carboxylic acid deno-3- carboxyloxymethyl-7-methoxy-3-cephem-4carboxylic acid di-benzhydryl ester 1.1 g, 1.18 mmol , acetonitrile 5 ml and bis-trimethylsilyl trifluoroacetamide 3 ml are left to stand at room temperature for 6 hours. After this period, the volatile products were removed under high vacuum and the residue was dissolved in 3 ml of methylene chloride. To this solution was added phenylacetyl chloride 0.23 ml, 1.79 mmol and the mixture was left to stand at room temperature for 65 hours. The solution was then evaporated and the residue dissolved in 5 ml tetrahydrofuran and 0.7 ml 2.5 N hydrochloric acid. acid. After 20 min of reaction time, the solvent was evaporated and the residue partitioned between methylene chloride and sodium bicarbonate solution. The organic layer was washed with sodium chloride solution, dried and evaporated to dryness. The crude product obtained was purified by chromatography on silica gel using chloroformethyl acetate 95: 5 as eluent. The title compound was obtained homogeneously by TC chromatography.

UV: / CH ₃ OH / λ max. 2640 pm ε 6650;

NMR: (solvent - SDS1 ₃ / δ = 3.50 / O-CH ₃ , singlet), 3.31 (2-H ₂ , 9ci ₃ partially visible), 4.67 (CH ₂ ~ O, singlet), 5 , 04 (6-H, singlet), - 4.96 (10H ₂ , partially visible), 3.95 (13-H ₂ , singlet).

Step D. 3carbamoyloxymethyl-7-methoxy-7-phenylacetamido-3-cephem-4-carboxylic acid benzhydryl ester.

To 7p - // O-5'-Trichloroethoxycarbonylomino-5'-carboxyvaleryl-phenylacetylamino-3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid solution-dibenzhydryl ester 104 mg in 90% acetic acid acid-water 1 ml was stirred with 100 mg zinc powder for 5 hours. The solution was then filtered and the solvent removed in vacuo. The residue was partitioned between methylene chloride and water, and the methylene chloride layer was extracted with sodium bicarbonate and sodium chloride solutions. Drying and evaporation gave a crude product which was purified by flash chromatography, using silica gel plates and a 3: 2 mixture of chloroform and ethyl acetate. The product is characterized by IR and NMR spectra. IR: / CHC1 _3/1780, 1730 and 1680 cm ^1. UV: / CH ₃ OH / λ max. 2640 pm ε 5870; NMR: (Solvent -CDCI ₃ / δ = 3.40 / O-CH ₃ , S), 3.33 (2-H ₂ , partially visible), 5.01 (6-H), 4.88 / 10- H2 is partially visible /,

3.60 (13-H ₂ , singlet).

Step E. 3-Carbamoyloxymethyl7-methoxy-p-phenylacetamido-3-cephem-4-carboxylic acid

3-Carbamoyloxymethyl-7-methoxy-7H-phenylacetamido-3-cefrem-4-carboxylic acid benzhydryl ester 17 mg was dissolved in anisole 0.2 ml, and treated with trifluoroacetic acid 0.5 mg for 5 min. After this period, the mixture was concentrated rapidly in high vacuum and diluted with ethyl acetate. The product was removed from the ethyl acetate solution by extraction with sodium phosphate buffer pH 7.5. The buffer solution was acidified to pH 2 with dilute hydrochloric acid and the title compound was separated by extraction with ethyl acetate. The product is obtained after drying and evaporating the solution. An analytical sample was obtained by recrystallization from ethyl acetate. Melting point 159-161 ° C. UV: / Buffer with pH 7 / λ max. 2670 µm ε 8650; IR: / CH ₃ Cl / 1780, 1735 and 1700; NMR: (solvent - CD ₃ CN + D ₂ O / δ = 3.42 / O-CH ₃ / s), 3.35 (2-H ₂ , partially visible), 5.01 / 6-H, s / , 4.83 / 10-H ₂ / d / 3.61 / 13-H ₂ /. Elemental analysis for C _lg H ₁₉ -O _? N ₃ S. Calculated: C, 51.29; H, 4.54. Found: C, 51.74; H, 7.73.

Two milligrams of the above acid was dissolved in one drop of methanol and treated with a solution of 2 mg dibenzylethylenediamine diacetate in ethyl acetate. The dibenzylethylenediamine salt of the title compound was precipitated after standing in the form of needle crystals. Melting point 140 ° C-143 ° C. UV: / CH ₃ OH / λ max. 263 mm ε 8600.

Preparation of the monosodium salt of 7β- (D-5-amino-5-cyclopentylamino) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid.

Modified fermentation process

Stage 1. Sloping crops

Lyophilized culture vial of Streptomyces lactamduraus / MA125

2908 / is opened aseptically and the organism is transferred to an environment with the following composition:

Medium XI% molasses% yeast from National Brewer 2.5% Difco agar pH 7.0 Water to volume

The inclined crops were inoculated for seven days at 28 ° C. When stored in the cold, they are stable for more than 13 weeks.

Stage 2. Stages of sowing. Two-stage system

First sowing. The first crop was inoculated directly from the inclined culture from stage 1 in 40 ml of 1% primary dried yeast, NF, pH 7.0 (supplied by Yeast Product Corporation) in a 250 ml narrow-necked Erlenmeyer flask. The flasks are then shaken on a rotary shaker at 220 rpm with an amplitude of 5.08 cm for 2 to 3 days.

Second crop. 2.5% inoculum from the first stage of sowing was added to a flask containing 2% Fleishmann S-150 yeast autolysate, pH 7.0. Growth in this stage is typically mild and incubation, carried out as in the first stage, lasts more than 40 hours.

Stage 3. Production environment

The trace product contains 30 g of distillate soluble matter per liter of distilled water; 7.5 g of NG primary yeast and 0.25% v / v Mobilpar-S antifoam. The medium was brought to pH 7.0 with a small amount of concentrated sodium hydroxide solution, partitioned in Erlenmeyer flasks and autoclaved for 15-20 min at 121 ° C. After cooling, a 2.5% inoculum is obtained from the culture obtained in step 2. The incubation time may vary from about 50 h to 100 h, but an incubation period of 72 h is preferred. The volume of the medium in each flask may vary 30 to 50 ml but 40 ml are routinely used. The inoculum concentration may vary from 1 to 5%, but in practice a concentration of 2.5% is generally used.

Stage 4. Determination

When fermentation is complete, cells are separated by centrifugation and the culture medium is diluted with phosphate buffer pH 7.0. The concentration of 7β- (O-5-amino-5-carboxyvaleramido / -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid in a fermentation medium was determined by a standard biological disk test method. The test organism used is Vibrio percolans (ATCC 8461). Filter paper disks are immersed in diluted culture media and placed on the surface of Petri dishes containing agar, which have been inoculated with the test organism Yibrio percolans (ATCC 8461). These petri dishes also contain disks that have been previously immersed in a standard solution containing known concentrations of 7β- / O-5-amino-5-carboxyvaleramido / -3-carbamoyloxymethyl-7-methoxy-3-cefem-4carboxyl acid. The disks were incubated overnight at 28 ° C and the diameter of the inhibition zones recorded. The concentration of the product and the fermentation medium were calculated by interpolation from a standard curve which gave the ratio of the diameter of the area to the known concentrations of the standard solutions of the product. By this method, Streptomyces lactamdurans MB-2908 is calculated to produce 78.6 mg / ml 7β / O-5-amino-5-carboxyvaleramido / 3-carbamoyloxymethyl-7-methoxy-3cephem-4-carboxylic acid by the mo126-modified fermentation process . Step 5. Isolation The filtered medium was adjusted to pH 7.0 with dilute hydrochloric acid and 2900 ml was passed through a column containing a highly basic anion exchange resin 100 g having a styrene-vinylbenzene matrix Dowex 1 x 2 chloride cycle, resin with rate 10 ml / min. The spent solvent was collected in 500 ml fractions. The resin column was washed with water and then eluted with 3% ammonium chloride in 90% methanol. The eluate was collected in 100 ml fractions. The eluted fractions were combined, the pH was adjusted to 7.2-8.0 with dilute sodium hydroxide and absorbed on a highly basic anion exchange resin 100 g having a styrene-divinylbenzene matrix Dewex 1 x 2 chloride cycle and a resin with 14 ml / min . The column was washed with water and eluted with 5% aqueous sodium chloride solution. The eluate was collected into 50 ml fractions and concentrated. The concentrate is diluted to 500 ml, adjusted from pH 8.8 to pH 2.0 with dilute hydrochloric acid and adsorbed onto 25 ml of a highly acidic sulfonate type cation exchange resin having a styrendinylbenzene matrix, Dowex with 50 x 2 hydrogen resin cycle, at

2.5 ml / min. The column was washed with 25 ml of water, then eluted with 2% pyridine until the pH of the column current increased to pH 7 54 ml. The resulting eluate was adjusted to pH 8.0 with dilute sodium hydroxide and concentrated in vacuo to remove the pyridine to give the 7-p-O-5-amino-5-carboxyvaleramido / -3-carbamoyloxymethyl-7 monosodium salt. -methoxy-3-cephem-4-carboxylic acid.

Elemental analysis for C ₁₆ H ₂₁ N ₄ S0 ₉ Na. Calculated: 41%; H, 4.5%; N

12.0%; With 6.8%. Found: C, 39.31%;

H, 4.76%; N, 11.16%; With 6.46%.

Example 169

3-carbamoyloxymethyl-7-methoxy-p-phenylacetamido-3-cephem-4-carboxylic acid

Step A. 7p- (O-5'-Trichloroethoxycarbonylamino-5'-carboxyvaleryl / phenylacetamido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid di-benzhydryl ester

7p- (O-5-Trichloroethoxycarbonylamino-5-carboxyvaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid dibenzhydryl ester solution 9.3 g, 10 mol, N-trimethylsilylphthalimide 7.8 g , 40 mol, phenylacetyl chloride 5.3 ml, 40 mol, in 50 ml of acetonitrile was heated to 40 ° C for 20 h. After this period, the mixture was cooled to room temperature and filtered. The filtrate was evaporated to dryness and triturated with hexane. The insoluble residue containing 7 N- (N-5'-trichloroethoxycarbonylamino-5'-carboxyvaleryl / phenylacetamido) -3-carbamoyloxymethyl-7methoxy-3-cephem-4-carboxylic acid di-benzhydryl ester was used without purification.

Step C. 3-Carbamoyloxymethyl-7-methoxy-7-phenylacetamido-3-cephem-4-carboxylic acid benzhydryl ester

The crude product from step A was dissolved in a mixture of ethyl acetate 50 ml, acetic acid 45 ml and water 5 ml. To this solution was added 20 g of zinc powder and the mixture was shaken at room temperature for 4 h. The excess zinc was then removed by filtration and the filtrate was partitioned between ethyl acetate and water. The organic layer was washed with brine and water, dried, and the solvent was evaporated. The crude product obtained was purified by chromatography on 1 kg of silica gel using a mixture of chloroform, hexane and methanol 47: 47: 6 for elution. The product thus obtained has the physical characteristics described in Example 168, step E.

Step C. 3-Carbamoyloxymethyl7-methoxy-phenylacetamido-3-cephem4 ~ carboxylic acid

The title compound was prepared according to the procedure described in Example 168, step F and had the same physical characteristics as the product of Example 168.

Example 170

3-carbamoyloxymethyl-7-methoxy-7- (2-thienylacetamide) -3-cephem-4-carboxylic acid

Step A. 7P- (O-5'-Trichloroethoxycarbonylamino-5'-carboxyvaleryl) -2-thienylacetylamino / -3-carbamoyloxymethyl7-methoxy-3-cephem-4-carboxylic acid di-benzhydryl ester

Mixture of 6.0 g, 6.3 mol, 7b- (O-5'-trichloroethoxycarbonylamino-5'-carboxyvaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid dibenzhydryl ester, 4, 7 g, 40 mmol, N-trimethylsilyl trifluoroacetamide, 3.42 ml, 25 mmol,

2-Thienylacetyl chloride and 50 ml of chloroform were heated at 47 ° C for 16 h. After the solvent was removed by evaporation, the crude reaction mixture was extracted with hexane and further purified by chromatography on 1 kg of silica gel using eluent 10% ethyl acetate in chloroform. HC / methanol / λ max. 265 mm is 5810; NMR: (solvent-deuterium chloroform (d = 3.53) -OCH ₃ , singlet), ~ 3.4 (2-H doublet), 4.74 (CH-O, syn-).

X II X glet /, 5.05 (6-H, singlet), 5.0 / 10-H ₂ , partially visible /, 4.15 (13-H ₂ , singlet).

Stage B. Benzhydryl ester of

3-Cabamoyloxymethyl-7-methoxy-7P (2-thienylacetamido) -3-cephem-4-carboxylic acid

7β / N-5'-Trichloroethoxycarbonylamino 5'-carboxyvaleryl / -3-thienylacetylamino-3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid di-benzhydryl ester 4.2 g, 3.8 mol, was dissolved in 30 ml of ethyl acetate and 30 ml of 90% aqueous acetic acid and 12 g of zinc powder are added. The mixture was stirred vigorously for 5 1/2 hours at room temperature. After the zinc is filtered off, the excess acetic acid is removed by washing the ethyl acetate solution with water. The title compound was isolated in the same manner as described in Example 1, Step E. It was characterized by thin layer chromatography (7% CH ₃ OH in 1: 1 CHCl ₃ : n-hexane) as a single spot material .

IR (CHCl ₃ ), 1740, 1800 cm ¹ ;

UV λ 263 gm ε 5800

NMR (solvent - CDCI ₃ (δ = 3.45) -OCH ₃ , singlet), ~ 3.4 (2-H ₂ , doublet), 5.02 (6-H, singlet), ~ 4.92 / 10 -H ₂ , partially visible (3.85 / 13H ₂ , singlet).

Step C. 3-Cabamoyloxymethyl-7-methoxy-7β- (2-thienylacetamido) -3-cephem-4-carboxylic acid

Cold solution of 3-cabamoyloxymethyl-7-methoxy-7β- (2-thienylacetamido) -3-cephem-4-carboxylic acid benzhydryl ester 1.36 g in 10.88 ml of anisole was stirred with 5.44 ml of trifluoroacetic acid 1/2 h. The volatiles were removed under high vacuum and the product was recrystallized from ethyl acetate. Melting point 165-167 ° C.

128

263μπι ε8844 UV / pH 7 buffer / λ _Μ3κε

263µηι ε 1400 / α / ₀ / C = 1, CH ₃ OH / = + 199 ° C

NMR: (solvent-CD ₃ CM + D ₂ O (δ = 3.48) -OCH ₃ , singlet), -3.4 (2H ₂ , partially visible), 5.05 (6-H, singlet), 4 , 91 (10-H ₂ , doublet), 3.86 (13H ₂ , singlet).

Elemental analysis for C ₁₆ H ₁₇ N ₃ O _? S ₂ . Calculated: C, 44.96; H, 4.01; N, 9.83. Found: C, 44.86; H, 3.99; N, 9.21; S 15.00.

Step D. 3-Carbamoyloxymethyl-7-methoxy-7β- (2-thienylacetamido) -3-cephem-4-carboxylic acid sodium salt

A suspension of 1 g of 3-cabamoyloxymethyl-7-methoxy-7β- (2-thienylacetamido) -3-cefrem-4-carboxylic acid in 100 ml of distilled water was stirred at room temperature, and 0.2 g was stirred gradient. After a solution was obtained and the pH was practically neutral (pH 6-7), the portion was filtered into a lyophilization vessel. The filtrate was lyophilized.

1 g of amorphous sodium salt of 3-carbamoyloxymethyl-7-methoxy-7P- (2-thienylacetamido) -3-cephem-

4-carboxylic acid corresponding to 99% recovery. UV (pH 7 buffer): E% 198 at 262 nm, 315 at 236 ps. IR (KBr): 1760 (lactam). (a / _D = 183.1 ° C / C = 1, pH 7 buffer).

Example 171

3-Acetylaminoacimethyl-7β- (2-pherylacetamido) -7-methoxy-3-cephem-4 carboxylic acid

Step A. 7β- / D-5'-Ethoxyphenylamino-5'-carboxyvaleryl / -2-furylacetylamino / -3-carbamoyloxymethyl-7methoxy-3-cephem-4-carboxylic acid dibenzhydryl ester

A mixture of dibenzhidrilov ester of 7β- / D-5'-τpixlopeτoκci carbon ylamino-5'-karboksivalerilamido / -3karbamoiloksimetil-7-methoxy-3tsefem-4-carboxylic acid 9,3 g, Bis / trimethylsilyl / -trifluoratsetamid 7,0 ml , 2-furylacetyl chloride 4.7 ml and dichloromethane 50 ml were heated at 47 ° C for 16 h. The solvent was evaporated, the crude reaction mixture was extracted with hexane and the residue was used in the next step without purification.

NMR: (solvent - SDS1 ₃ / δ = 3.48 / -OCH ₃ , singlet), 3.08 / 2-H ₂ , ςα doublet /, 4.63 (CH ₂ , singlet), 5.02 / 6- H, singlet /; ~ 4.88 / 10-H ₂ , doublet /, 3.72 / 13-H ₂ , singlet /.

Step B. 7β / 2-furylacetamido-3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid benzhydryl ester

Step A dibenzhydryl ester is reacted with zinc powder and acetic acid, following the procedure described in Example 170, step B. After crystallization from chloroformhexane, the pure product has the following characteristics:

Melting point 168-71 ° C.

IR / CHC13 / 1800, 1720, 1700 UV λ max. 265 μιτι ε 7200

NMR: / solvent - CD ₃ Cl / δ =

3.43 (-OCH ₃ , singlet), 3.39 (2-H ₂ , partially visible), 5.0 (6-H, singlet),

4.75 (10-H ₂ , doublet), 3.64 (13-H ₂ , singlet).

Step C. 3-Carbamoyloxymethyl-7-methoxy-N- (2-furylacetamido) -3-cephem-4-carboxylic acid

3-Carbamoyloxymethyl-7-methoxy7β- (2-phenylacetamido / -3-cephem-4 carboxylic acid is obtained from the product of step B, following the procedure described in example 170, step C. The product after crystallization from ethyl acetate has a point of mp 156-161 ° C.

UV / pH buffer 7 / λ max. 265 μπι ε 7200

The IR is the same as the structure

NMR: (solvent -CD ₃ CN + D ₂ O / δ = 3.44) -OCH ₃ , singlet / - 3.38 (2-H ₂ , partially visible), 5.02 / 6-H, singlet /, 4 , 82/10-H ₂ , doublet /, 3.66 / 13-H ₂ , singlet /.

Example 172

3-carbamoyloxymethyl-7-methoxy7P-thiophenoxyacetamido-3-cephem-4 carboxylic acid

Step A. 7P- (R-5-Trichloroethoxycarbonylamino-5-carboxyvaleryl) -thiophenoxyamido] -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid dibenzhydryl ester

Following the procedure described in Example 171, step A, and instead

2- furilatsetilhlorid using an equivalent amount feniltioatsetilhlorid was prepared dibenzhidrilov ester 7β - // D-5-trihloretoksikarbonilamino-5-karboksivaleril / -tiofenoksiamido / -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid.

NMR: (solvent -CHCl ₃ (δ = 3.33) -OCH ₃ , singlet), ~ 3.23 (2-H ₂ , ςα partially visible), 4.87 (CH ₂ ~ O-, and /, 5 , 0 (6-H, singlet), 4.87 (10-H ₂ , and), 3.68 (13-H ₂ , singlet).

Step B. 3bapachamoyl ester-7-methoic-7βthiophenylacetamido-3-cefem-4-carboxylic acid benzhydryl ester

Following the procedure described in Example 171, step B, using 7β - // ϋ- dibenzhydryl ester.

5-Trichloroethoxycarbonylamino-5carboxyvaleryl (-thiophenoxyamido) -

3- Carbamoyloxymethyl-7-methoxy-3cephem-4-carboxylic acid in place of 7β - [(5'-trichloroethoxycarbonylamino-5'carboxyvaleryl) -2-furylacetamino] -3-carbamoyloxymethyl-7-methoxy-3-methoxy-3-methoxy-3- carboxylic acid, after chromatographic purification, a pure product is obtained which manifests as a single spot on a thin layer chromatogram / TLC. The IR corresponds to the structure. UV: λ max.274 μιη ε 11350. NMR: / solvent - CDCI ₃ / δ = 3.34 / -OCH ₃ , singlet /, 3.24 / 2-H ₂ , partially visible /, 5.0 / 6- H, singlet /,

4.88 (10-H ₂ , doublet), 3.68 (13 ~ H ₂ , singlet).

Step C. 3-Carbamoyloxymethyl-7ΜετοκοΗ-7β-τποφετοκοπ3πετΒΜΗΛθ-3cephem-4-carboxylic acid

Said compound is prepared from the product of step B above according to the procedure of Example 170 step C. The product has one TLC stain. Melting point 119-123 ° C.

UV / pH buffer 7 / λ max. 247 gm ε 10400.

NMR: (solvent CD ₃ CI + D ₂ O / δ = 3.38 (OCH ₃ , singlet), 3.34 (2-H ₂ , partially visible), 5.0 / 6-H, singlet /, 4, 82/10-H ₂ , singlet /, 3.71 / 13 ~ H ₂ , singlet /.

Example 173 β- N-L-a-a Zidophenic Lacetas Mido / 3-Carbamoyloxymethyl-7-methoxy-3cephem-4-carboxylic acid

Step A. 7β- (N-5-tert-6-benzylamino-5-carboxyvaleramido / thiophenoxyamido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid. 7β- [ϋ-5-3ΜΗΗθ-5-S-3-boxyvaleramido / -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid 50 g was dissolved in a mixture of 1500 ml aqueous 5% dicalium hydrogen phosphate and 1000 ml acetone and adjust to pH 9.5 with 2.5 N sodium chloride solution. To this solution was added with stirring

130 tert -butoxycarbonylazide 50 ml and the pH was maintained at 9.5 for a period of 20

The reaction mixture is then extracted with ethyl acetate, the ethyl acetate layer is discarded, the aqueous layer is cooled to 0 ° C, stirred with 1200 ml ethyl acetate and acidified to pH

2.5 with concentrated hydrochloric acid. The ethyl acetate layer was separated, dried over sodium sulfate and concentrated in vacuo, and the solid thus obtained could be used without further purification.

IR 1790 (β-lactam), 1700

UV / pH 7 buffer / λ max.263 gm ε 6820

NMR: (solvent -DMSO, d ₆ (δ = 3.30) -OCH ₃ , singlet), 3.42 (2H ₂ , partially visible), 5.06 (6-H, singlet), 4.78 / 10 -H ₂ , doublet /, 1.38 / tert.You, singlet /.

Step B. Dibenzhydryl ester of 7p- / O-5-tert-butoxycarbonylamino-

5-Carboxyvaleramido / -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid

To a solution of 7β- / D-5-butoxycarbonylamino-5-carboxyvaleramido / -3-carbamoyloxymethyl-7-methoxy3-cephem-4-carboxylic acid 15 g in ethyl acetate 500 ml was added diphenyldiazomethane 5.5 g in 70 ml ether. The reaction mixture was warmed to 40 ° C with stirring and after 30 min treated with additional diphenyldiazomethane 5.5 g in ether 70 ml. After 3 h, the solvent was removed in vacuo and replaced with a mixture of methanol 500 ml and water 20 ml. The methanol-water solution was extracted 4 times with hexane and then evaporated in vacuo. The residue was dissolved in ethyl acetate and then evaporated in vacuo. The residue was dissolved in ethyl acetate, dried over sodium sulfate and evaporated in vacuo. The title compound is obtained which is used without purification in the next step.

NMR: / solvent -CDCI ₃ / δ =

3.60 (-OCH ₃ , singlet), 3.4 (2-H ₂ , partially visible), 5.10 (6-H, singlet), 4.95 (10-H, partially visible).

Step C. 7β- (D-5'-tert.-Butoxybenzylamino-5'-carboxyvaleryl) -D, L-α-azidophenylacetamino / -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid-4-carboxylic acid

Mixture of 7β- (D-5'-methyl-5-carboxyvaleramido) -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid dibenzhydryl ester 10.8 g, chloroform 100 ml, bis-bis N-Trifluoroacetamide 16.2 g and D, L-α-azido-phenylacetyl chloride was heated to 45 ° C for 16 h. The mixture was diluted with chloroform 300 ml, washed with 2% aqueous bicarbonate and saturated sodium chloride solution, dried was concentrated over sodium sulfate and evaporated to an oil which was purified by precipitation of the product from a chloroform solution with hexane. The pale yellow solid was used in the next step without further purification.

IR 1790 / β-lactam /, 1735, 2100 / -N, /

NMR: (solvent CDCI ₃ / δ = 3.70 / -OCH ₃ , singlet), 3.2 (2-H ₂ , partially visible)

Step D. 7β- N, L-α-Azidophenylacetamido / -3-carbamoyloxymethyl-7 methoxy-3-cephem-4-carboxylic acid

7β- (D-5'-tetoperoxybenzylamino-5'-carboxyvaleryl) -D, L-a-azidophenylacetamino / -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid-4-carboxylic acid solution of dibenzhydryl ester Pour 0 g into anisole 13 ml onto 65 ml of cold (0 ° C) trifluoroacetic acid. After 5 min, 131 the product was poured into 1800 ml of stirred, cold 0 ° C ether. The precipitated solid was collected and partitioned between 10% aqueous disodium acid phosphate and ethyl acetate. 5 The ethyl acetate layer is discarded and the aqueous layer is covered with fresh ethyl acetate and the stirred mixture is brought to pH 2 with cold 60% aqueous phosphoric acid solution. 10 The ethyl acetate layer was collected, washed with saturated aqueous sodium chloride solution and dried over sodium sulfate. The light fractions were removed in vacuo. The compound indicated in the title was obtained.

264gm ε 7537 / pH 7 buffer / UV λ max.

231gm ε 13567

IR 1760 / β-lactam / 1705, 2105 / -Ν ₃ /. 20 NMR: solvent CD ₃ CN / δ = 3.36 / ~ OCH ₃ , singlet /, 3.50 / ~ OCH ₃ , singlet /, 3.40 (2-H ₂ , partially visible), 5.06 / 6 -H, singlet /, 4.86 (10-H, singlet), 5.15 (13-H, singlet). 25

Example 174 7β - / - N, L-α-azidophenylacetamido / -3-carbamoyloxymethyl-7-methoxy3-cefem-4-carboxylic acid

A suspension of 1.0 g of 7β - / - ϋ, L-α-30 azidophenylacetamino / -3-carbamoyloxymethyl-7-methoxy-3-cephem-4-carboxylic acid in acetic acid 10 ml and water 90 ml at 0 ° C stirred with zinc powder 5.0 g for 10 min 35 and filtered. The filtrate was sprayed with hydrogen sulfide, filtered and the filtrate was freeze-dried. A white solid was obtained, which was washed with ether and dried under 40 vacuo to give the title compound as a white powder.

In B / pH 7 buffer / λ max.264 gm ε 6525

IR 1770 / β-lactam / 2650, 1550 45 / ΗΝ ₃ + /

NMR: / solvent D ₂ O + HCO ₃ /

6 = 3.78 (~ OCH ₃ , singlet), 3.84 (-OCH ₃ , singlet), 3.90 (2-H ₂ , partially visible).

Example 175

3-Acetoxyacetyl-7β- (2-thionylacetamide) -3-cephem-4-carboxylic acid

Step A. 7β- / D-5-Tropyloxycarbonylamino-5-carboxyvaleramido / -3-acetylmethyl-3-cephem-4-carboxylic acid

To a solution of 7β- (N-5-O-5-carboxyvaleramido) -3-acetoxymethyl-3-cephem-4-carboxylic acid

2.5 g, 0.53 mol in acetone 13 ml and aqueous 10% solution of dicalium hydrogen phosphate 40 ml were added dropwise trichloroethoxycarbonyl chloride 3.35 g, 0.159 mol. During the addition, the pH of the solution was maintained within the range of 8.5 to 9 by the gradual addition of 17% aqueous sodium hydroxide solution. After 30 min, the mixture was washed with ethyl acetate and the aqueous layer was acidified to pH 2.5 with concentrated hydrochloric acid. The precipitated product was extracted with ethyl acetate, the solution was dried over sodium sulfate, filtered and the solvent was removed. 2.7 g of 7β (O-5-trichloroethoxycarbonylamino-5carboxyvaleramido) -3-acetylmethyl 3-cephem-4-carboxylic acid are obtained.

Step B. Dibenzhydryl ester of 7- (O-5-trichloroethoxycarbonylamino-

5-Carboxyvaleramido / -3-acetoxymethyl-3-cephem-4-carboxylic acid

To a solution of 7β- (D-5-pyrroloxy-carbonylamino-5-carboxyvaleramido) -3-acetylmethyl-3-cephem-4-carboxylic acid in ethyl acetate 30 ml was added diphenyl diazomethane 2.0 g in ether 25 ml. The mixture was stirred overnight and the solvent removed to give 4.0 g of crude product. The crude product was purified by chromatography on silica gel using chloroform

132 for eluent to give 2.3 g of substantially pure 7- (O-5-trichloroethoxycarbonylamino-5-carboxyvaleramido) -3-acetylmethyl-3-cephem-4-carboxylic acid dibenzhydryl ester.

NMR: (solvent-SDSC / δ = 2.0 (methyl, singlet), 4.9 (10-H ₂ , quartet), 3.2 (2-H ₂ , quartet), 4.95 / 6-H, doublet /, 5.92 (7-H), 7.0 (benzhydryl protons, 2 singlets).

Step C. 7 - // D-5-Trichloroethoxycarbonylamino-5-carboxyvaleryl / -2-thienylacetamino / -3-acetoxymethyl-3-cephem-4 carboxylic acid dibenzhydryl ester

Mixture of 7P- / O-5-trichloroethoxycarbonylamino-5-carboxyvaleramido / -3-acetoxymethyl-3-cephem-4-carboxylic acid dibenzhydryl ester 2.0 g, 0.02 mol, N-trimethylsilyl trifluoroacetamide 1.65 g, 0.09 mol, 2-thienylacetyl chloride 1.31 g, 0.0815 mol, and methylene chloride 6 ml were heated to 40-45 ° C in an oil bath under a nitrogen atmosphere for 20 h. The reaction mixture was poured onto hexane 100 ml and poured into hexane. filtered through diatomaceous earth. Removal of the solvent gave 7 // O-5-trichloroethoxycarbonylamino-5-carboxyvaleryl (-2-thienylacetamino) -3-acetoxymethyl-3-cephem-4-carboxylic acid dibenzhydryl ester.

Step D. 3-Acetoxymethyl-7- (2-thienylacetamido) -3-cephem-4-carboxylic acid benzhydryl ester

Dibenzhydryl ester of 7 // D-

5-Trichloroethoxycarbonylamino-5Carboxyvaleryl (-2-thienylacetamino) -3-acetoxymethyl-3-cephem-4carboxylic acid was dissolved with ethyl acetate 10 ml and added to a mixture of 90% aqueous acetic acid 10 ml and zinc powder 1.0 g. The mixture was stirred for 2 h at room temperature. The reaction mixture was filtered to remove zinc. The reaction mixture was washed successively with 2 portions of water, cold sodium bicarbonate solution and then with saturated sodium chloride solution 15 ml. The ethyl acetate solution was dried over sodium sulfate, filtered and the solvent was evaporated. It turns out

1.9 crude product which was chromatographed on silica gel using a mixture of chloroform and ethyl acetate as eluent 50: 1. 0.380 g of product is obtained which, after recrystallization from ethyl acetate, has a melting point

141.5-143 ° C.

HC / CH ₃ OH / λ max. 263 δ 7580

Elemental analysis for C, _o H ,, _{N n} 0, S ,. Calculated C, 61.91; H, 4.66; N, 4.98; Found C, 62.14; H, 4.84; N, 4.91.

Step E. 3-Acetoxymethyl] -7- (2-thienylacetamido) -3-cephem-4-carboxylic acid

A cold solution of 3-acetoxymethyl-7- (2-thienylacetamido) -3-cephem-4-carboxylic acid benzhydryl ester 100 mg in anisole 1.0 ml and trifluoroacetic acid 0.5 ml were stirred at 0 ° C for 35 min. Tetrachloromethane 50 ml was added and the reaction mixture was concentrated to dryness. The residue was treated with hexane. The hexane was removed by decantation and the residue dissolved in ethyl acetate 10 ml, concentrated to 1 ml and diethyl ether added to give a precipitate. This precipitate was crystallized from a mixture of diethyl ether and ethyl acetate. 0.025 g of 3- (acetoxymethyl) -7- (2-thienylacetamido) -3-cephem-4-carboxylic acid are obtained with a melting point of 164 ° C. The combined melting point with the original sample was 163 ° C.

Example 176

3-Methyl-7-methoxy-7-N- (2-thienylacetamido) -3-cephem-4-carboxylic acid dibenzylethylenediamine salt

133

Step A. 7-N- (O-5-Amino-5-carboxyvaleramido) -3-methyl-7-methoxy-3-cephem-4-carboxylic acid

The 10% palladium on carbon catalyst was suspended in water for 10 ml and treated with hydrogen. The catalyst was filtered off and resuspended in water for 50 ml and to this mixture (2.67 g) was added the 7-P- (O-5-amino-5-carboxyvaleramido / -3-carbamoyloxymethyl-7-methoxy) sodium salt. cefem-4-carboxylic acid 1.0 g in water 10 ml. The resulting mixture was shaken for 22 h at room temperature. The catalyst was removed by filtration and washed with 50 ml of water. The combined wash water and filtrate were concentrated to dryness, yielding a 52.8% yield of 7-P- / O-5-amino-

5-Carboxyvaleramido / -3-methyl-7-methoxy-3-cephem-4-carboxylic acid, 528 mg. HC λ max. 265 µm; The _cost is 100.

Step B. 7b- (0-5-tert-Butoxycarbonylamino-5-carboxyvaleramido) -7methoxy-3-methyl-3-cephem-4-carboxylic acid dibenzylethylenediamine salt

7β (O-5-Amino-5-carboxyvaleramido) -7-methoxy-3-methyl-3-cephem-4-carboxylic acid solution of dβ and sodium salt 11.5 g were dissolved in water 150 ml and acetone 50 ml. The pH was adjusted to 9-9.1 with sodium hydroxide and 10 ml of tert-butyl azide formate were added. The reaction mixture was stirred for 16 h at room temperature and additional sodium hydroxide was added to maintain pH 9-9.1. The reaction mixture was extracted with ethyl acetate 100 ml and the organic layer was removed. The product was precipitated by lowering the pH to 2.5 with dilute hydrochloric acid. The precipitate was collected by centrifugation and converted to its dibenzylethylenediamine salt, which was crystallized from ethyl acetate. It works

4.3 g of 73- (O-5-tert-butoxycarbonylamino-5-carboxyvaleramido) -7-methoxy-3-methyl-3-cephem-4-carboxylic acid dibenzylethylenediamine salt, mp 177 ~ 179 ° C ( with disintegration). HC λ max. 263 μπι, 238 E, = 98.2, 81.1. Elemental analysis for C ₃₆ H ₄₉ N ₅ O ₉ S. Calculated: C, 59.42; H, 6.74; N, 9.63; Found: C, 60.02; H, 6.80; N, 9.79.

Step C. Dibenzylethylenediamine salt of 3-ΜετΗΠ-7-ΜετοκεΗ-7β- / 2-phenylacetamide-3-cephem-4-carboxylic acid

7β- / D-5-methylbenzoylamino-5-carboxyvaleramido / -3-methyl-7-methoxy-3-cephem-4-carboxylic acid was treated with aqueous dilute hydrochloric acid 200 ml, 0.1 N, and ethyl acetate 100 ml to extract the free acid. To a solution of 1.33 g, 2.74 mmol of free acid in methylene chloride 10 ml was added bis-trimethylsilyl trifluoroacetamide 2.2 ml and mono-trimethylsilyl trifluoroacetamide 0.5 ml. 2-Thienylacetyl chloride 1.1 ml was then added and the reaction mixture was stirred for 18 h under nitrogen at 43 ° C. The solvent was removed in vacuo and the residue partitioned between ethyl acetate and aqueous phosphate buffer pH 7.5. The aqueous layer was acidified with dilute hydrochloric acid and the precipitated product was extracted with ethyl acetate. The addition of dibenzethylenediamine resulted in the crystallization of 250 mg of the desired product in the form of a salt in the ratio of 2 equivalents of the product per 1 mol of dibenzethylenediamine. Recrystallization of the ethanol salt affords a substantially pure product with a melting point of 153 ~ 155 ° C (decomposition) with a preliminary darkening.

Elemental analysis for C ₄₆ H ₅₂ S ₄ N ₆ O ₁₀ .

134

Calculated: C, 56.54; H, 5.36; N, 8.60; S, 13.12; Found: C, 55.76; H, 5.16; N, 8.37; S 12.16.

Example 177

7- (phenylacetyl-2-thienylacetyl) amino-3-acetoxymethyl-3-cephem-4-carboxylic acid

Suspension of sodium cephalothin

3.36 g in anhydrous, alcohol-free chloroform 20 ml was silylated by the addition of trimethylchlorosilane 2.2 ml. After stirring for 30 min, monosylyl trifluoroacetamide 5.0 ml and phenylacetyl chloride 4.0 ml were added and the mixture was then heated to 45 ° C for 2 days under a refrigerator equipped with a drying tube. The volatiles were evaporated to give a residue which was dissolved in 100 ml of ethyl acetate and washed 3 times with water. The ethyl acetate layer was dried over magnesium sulfate, filtered and the solution was evaporated in vacuo to a residue. The residue was triturated with chloroform, some insoluble matter was removed by filtration and the product was precipitated with hexane. This procedure is performed 2 more times. 7 (Phenylacetyl-2-thienylacetyl) amino-3-acetoxymethyl-3-cephem-4-carboxylic acid is obtained in a solvent-free form by freezing from a solution in benzene. IR / CHC1 _3/1780 μ, 1720 μ. NMR: (CDC1 ₃ ) corresponds to the structure; CH ₂ G == 0 (4.03 ppm), 2-thienyl-CH ₂ -C-4.3 ppm.

δ and Thin layer chromatography: One major spot cR _f = 0.69 (ethyl acetate-pyridine-acetic acid-water 62:21: 6: 11) on silica gel.

Example 178

7- (di-2-thienylacetyl) amino-3-acetoxymethyl-3-cephem-4-carboxylic acid

Suspension of sodium cephalothin

1.18 g in anhydrous, alcohol-free chloroform 10 ml was silylated by the addition of trimethylchlorosilane 1.1 ml. After stirring for 30 min, monosilyltrifluoroacetamide was added

2.5 ml and 2-thienylacetyl chloride 2.0 ml are added to the slurry, which is then heated to 45 ° C. The mixture was left at this temperature for 2 days and then evaporated in vacuo to a residue which was dissolved in ethyl acetate 50 ml and washed 3 times with water. The ethyl acetate layer was dried over magnesium sulfate, filtered and the filtrate was evaporated in vacuo to give a residue. The residue was dissolved in chloroform and precipitated with hexane 3 times, removing the supernatant fluid each time. 7- (di-2-thienylacetyl) amino-3-acetoxymethyl 3-cephem-4-carboxylic acid is obtained in a solvent-free form by freeze-drying from a solution in benzene.

NMR: (solvent - deuterium chloroform) - corresponds to the structure;

δ = 4.25 (2-thienyl // - CH ₂ ~ C, 4H); Thin layer chromatography: one major spot cR _f = 0.67 (ethyl acetate-pyridine-acetic acid62: 21: 6: 11) on silica gel.

Example 179

7- (2-thienyl) amino-3-acetoxymethyl-3-cephem-4-carboxylic acid

7- (Phenylacetyl-2-thienylacetamino-3-acetoxymethyl-3-cephem-4-carboxylic acid 250 mg was dissolved in tetrahydrofuran 10 ml and water 10 ml. The pH of the solution was adjusted to 9 and the mixture was allowed to stand for 1 hour. The solution was then extracted with ethyl acetate and the extracts washed with disodium hydrogen phosphate solution. After drying, the solvent is evaporated and a mixture of 7- (2-thienylacetyl) amino-3-acetoxymethyl-3-cephem-4-carboxylic acid and the starting material are left. The product was separated from the starting material by chromatography to give substantially pure 7/2-thienylacetyl / amino-3-acetoxymethyl-3-cephem-4-carboxylic acid. The ratio of these two products is 7: 3.

Example 180

According to the procedure described in Example 170, step A, and by replacing the thienylacetyl chloride described therein, an equivalent diester of 7diacylamino-3-carbamoyloxymethyl7-methoxy / or hydro-3-cephem-4-carboxylic acid is obtained with an equimolar amount of acylating agent. which, after removal of the N-protecting group in accordance with the procedure of Example 170, step B, and subsequent treatment by the procedure of step C, gives the desired

7-acylamino-3-cephem-4-carboxylic acid.

The following scheme and Table I illustrate the starting materials, intermediates and new end products that can be obtained by this new method. It should be noted that in the following scheme and table, compounds having 7-hydroxy substituent instead of 7-methoxy substituent are subjected to the same reaction to obtain compounds of Table I having 7-hydroxy substituent instead of 7-methoxy substituent.

OCH ₂ C1 ₃ o

II with I

NH oh

I II

HC ~ (CH ₂ ) ₃ s

I μ ^X OCH0 _{2 about}

VI

II c ^ I ΝΗ I

HC - (CH ₂ ) ₃ CN

I n ^X OCH0 _{2 about}

OCH ₂ C1 ₃

Fr.

II

CH ₂ OCNH ₂

C-OCHO ₂

II

IVb

136

Unblocking

---------->

agent

Oh

II

Unblocking

--------->

agent

St

Oh

II

CH ₂ OCNH ₂

Fr.

Id

R ¹ ^ 2 ^ - ° ^CH 2 ^_ ; O-CH ₂ CH ₂ -; S-CHz ; CH ₃ CH = CHCH- C = 0 II he 0 II

CH ₂ = CHSCH -; NOSSN / _CH2 / ₃ -;

TABLE 1

FOR ^CH '~' 0 ' ; but- £ ^ -sn ₂ - C ₂ H ₅ SCH ₂ - ; nC ₄ H, SCH ₂ - O-SN- ; θ-CH.CH-

nh ₂

nh ₂

137

ch ₂ nh ₂ nhso ₂ ch ₃

NH-C-NH.

II ²

NH

NH

II

NH-C-NH-

-OCH ₂ -;

CNCH ₂ -;

NH OCH, ^11D ~ \

NH-C-NHCH-f 2 ⁴ OCH ₃

N z \\

N N

II II

-CH ₃ -C - N-CH Example 181

7-amino-7-allyl-cephalosporanic acid benzhydryl ester

7-amino-7-benzyl-cephalosporanic acid benzhydryl ester

7- (Rnitrobenzylidinamino) -cephalosporanic acid benzhydryl ester was reacted with allyl chloride according to the procedure of Example 166 to obtain 7- (p-nitrobenzylidenamino) -7-allyl-cephalosporanic acid benzhydryl ester. It is then regenerated to 7-amino-7-allyl-cephalosporic acid benzhydryl ester using aniline hydrochloride.

7-Amino-7-benzyl-cephalosporanic acid benzhydryl ester was prepared in the same manner using benzyl chloride. 40

Example 182 7-Amino-7-carboxy-cephalosporic acid

7-amino-7-dithiocarboxy-cephalosporanoic acid benzhydryl ester

A solution of 0.5 g of 7 - / - p-nitrobenzylidenamino / cephalosporanic acid benzhydryl ester in 5 ml of tetrahydrofuran is obtained under a nitrogen atmosphere. Carbon dioxide is bubbled through the solution through bar25 until the color disappears. 50 ml of benzene was added and the solution was washed with pH 2 aqueous phosphate buffer. The benzene solution was dried with magnesium phosphate, filtered and evaporated. The product 7- (p-benzylidenamino) -7-carboxy-cephalosporanic acid benzhydryl ester was obtained.

Using carbon sulphide gas or dry ice (solid carbon dioxide) in the above reaction, the compounds 7- (p-nitrobenzylidenamino) -7-dithiocarboxy-cephalosporanic acid or 7- (p-nitrobenzylidenamino) -7carboxylate are obtained. cephalosporanate.

The amine moiety was regenerated using aniline hydrochloride to give the 7-amino-7-carboxy-dithiocarboxy-cephalosporanic acid benzhydryl ester compounds.

138

Example 183

7-amino-7-nitro-cephalosporanic acid benzhydryl ester

7- (p-nitrobenzyl benzyl ester) benzene ester and / or cephalic acid a new acid 571 mg was dissolved in 10 ml of dry acetonitrile containing 4 mg of acetone cyanhydrin nitrate. The latter has the formula:

CH ₃

I

Ο ₂ Ν - O —C —CN

I dream ₃

With vigorous stirring under nitrogen for 5 hours, a solution of diisopropylethylamine in 12 ml of acetonitrile was added. The reaction mixture was stirred overnight and the solvent removed in vacuo. The residue was stirred in 50 ml of benzene and washed successively with water, 1 M aqueous phosphate buffer pH 2, water and bicarbonate. After drying, filtration and removal of the solvent, the residue is a crude compound, which is suitable for further reaction without further purification. It can be purified, if desired, by chromatography on silica gel, eluting with 25: 1 chloroform-ethyl acetate. It is identified as 7- (p-nitrobenzylidenamino) -7-nitrocephalosporanic acid benzhydryl ester.

7- (p-Nitrobenzylidenamino) -7-nitroso-cephalosporanoic acid benzhydryl ester, 7- (p-nitrobenzylidenamino) -7-carbamoylcephalosporanoic acid 7-, p-carboxybenzoic acid (7-p-carboxybenzoic acid) 7- 7- (p-nitrobenzylidenamino) -7-sulfo-cephalosporanoic acid benzhydryl ester, 7- (p-nitrobenzylidenamino-7-sulfamoyl-cephalosporanoic acid benzyl) 7-, p-nitrobenzylidenamino-7-methyl-sulfonosulfonyl-7-sulfonosulfonyl-7-sulfonosulfonyl or benzhydride ov 7- / p-nitrobenzilidenamino / -7-phospho-cephalosporanic acid can be prepared using the appropriate reagents nitrosyl chloride, carbamoyl chloride, chloroformate, sulfamoyl or phosphate oxychloride.

The aforementioned imino derivatives may be regenerated to the amino function using either aniline hydrochloride or 2,4-dinitrophenylhydrazine as described below. The products obtained are, respectively, 7-amino-7-nitro cephalosporanic acid, 7-amino-7-nitrosocephalosporanoic acid, 7-amino-7carbamoylcephalosporanic acid, 7-amino-7-carboethoxy-cephalosporic acid, 7-amino-7-sulphosphoric acid , 7-amino-7-sulfamide cephalone challenges new acidic, 7-amino-7-methylsulfo-cephalosporic acid and 7-amino-7-phospho-cephalosporic acid. In all cases, the ester to be obtained is a benzhydryl ester.

Example 184

7-Aminocephalosporanate-B-oxide benzhydryl ester

7,500 aminocephalosporanic acid benzhydryl ester was dissolved in 10 ml of methylene chloride cooled to 0 ° C and treated with m-chloroperbenzoic acid 0.172 g for 1 h, during which time the reaction mixture was allowed to reach room temperature.

The reaction mixture was diluted with methylene chloride and washed with 5% NHCO ₃ solution three times and then with water, dried and evaporated to give the crude product. He

139 was stirred with 10 ml of ether for 1 h. The precipitate was filtered off and washed with ether to give the title product.

Similarly, the other 7-aminodecephalosporanic acid esters described herein can be converted to S-oxides. These oxides can be used as intermediates in the preparation of the sulfoxides given by formula I above. Alternatively, the various intermediates and end products described herein may be converted to the corresponding S-oxides following the procedure described above. The different ones

The S-oxide derivatives can be converted to the corresponding cephem compounds according to the method of the example that follows.

Example 185

3-Methoxymethyl-7-methoxy-7- (2-thienylacetamido / cephalosporanic acid benzhydryl ester, S-oxide obtained by oxidation of the corresponding cephalosporan with m-chloroperbenzoic acid 1.0 g, was dissolved in 10 ml of acetonitrile and 10 ml. dimethylformamide. Tin chloride equivalent and half acetyl chloride equivalent were added at 0 ° C. The mixture was stirred at 0 ° C for 1 h and then at room temperature for 1 h. The solvent was removed in vacuo and water was added. The mixture was extracted with ethyl acetate and the extract was washed with 0.1 NHCl, saturated sodium bicarbonate and water. After drying, the solvent was removed in vacuo to give 3-methoxymethyl-7-methoxy-7- (2-thienylacetamido) -cephalosporic acid benzhydryl ester.

The new cephalosporins of formula I are valuable antibiotics active against various gram-positive and gram-negative bacteria. Although in general their biological spectrum is similar to that of known cephalosporins, these new cephalosporins have some new and unexpected properties. They are active against many microorganisms that are resistant to known cephalosporins, such as cephaloridine and cephalothin, and are resistant to β-lactamase induced by cephalosporin-resistant clinical isolates of pathogens, such as E coli A. cloacal. They are more active against Proteus strains such as P. mirsbilis and are more active against Proteus morganii strains that are resistant to unsubstituted cephalosporins. Tse are useful for separating microorganisms from pharmaceutical, medical and dental equipment and as germicides for industrial applications, for example water based paints and circulating water in paper mills to inhibit the growth of harmful flora.

The 7-methoxycephalosporates obtained according to the method of the invention are generally more active than 7- (O-5'-amino-5'-carboxyvaleramido) -7-methoxycephalosporins against various Gram-negative organisms and have increased activity against positive organisms. For example, these 7-methoxycephalosporins are active against gram-positive pathogens such as Staphylococus aureus at a minimum inhibitory concentration (MIC) of about 1.5 mcg / ml, Streptococcus pyogenes at a MIC of about 0.7 mcg / ml and Diplococcus pneumoniae at an MIC of about 0.7 mcg / ml and against gram-negative organisms such as Aerobacter aerogenes at an MIC of about 3 mcg / ml Proteus vulgaris at an MIC of 1.5 mcg / ml and Proteus morganii at about 6 mcg / ml. The activities of the specific products of the foregoing examples are as follows: 3-carbamoyl 140 lloxymethyl-7-methoxy-7-phenylacetamido-3-cephem-4-carboxylic acid, S. pyogenes MIC 1.56 mcg / ml and P. vulgaris MIC 1.56 mcg / ml, 3-carbamoyloxymethyl-7-methoxy-7- (2-thienylacetamido-3-cephem-4-carboxylic acid, S. pyogenes MIC 0.78 mcg / ml and P. morgani MIC 12.5 mcg / ml; 3carbamoyloxymethyl-7-methoxy-7- (2-furylacetamido) -3-cephem-4-carboxylic acid, S. aureus MIC 6.25 mcg / ml and P. vulgaris MIC 1.56 mcg / ml; 3-carbamoyloxymethyl-7-methoxy-7-thiophenoxyacetamido-3-cephem-4-carboxylic acid, S. pyogenes MIC 0.78 mcg / ml and D. pneumoniae MIC 0.78 mcg / ml; 3-acetoxymethyl-7-methoxy-7- (2-thienylacetamido) -3-cefem-carboxylic acid, S. pyogenes MIC 1.56 mcg / ml and P. vulgaris MIC 0.78 mcg / ml and 3-pyridinmethyl7-methoxy -7- (2-thienylacetamido) -3cephem-4-carboxylic acid Serratia MIC 25 mcg / ml and S. areus MIC 156 mcg / ml.

The products obtained according to the invention can be used alone or in combination as an active ingredient for a wide variety of pharmaceutical preparations. These antibiotics and their corresponding salts may be used in capsule forms or tablets, powders or liquid solutions or as suspensions and elixirs. They can be taken orally, intravenously or intramuscularly. Suitable carriers that can be used in the formulations include, for example, mannitol, sucrose, glucose or sterile liquids such as water, saline, glycols and petroleum oils, oils of animal, vegetable or synthetic origin, for example peanut oil, mineral oil or sesame oil. . Also in addition to the carrier, a composition may include other ingredients such as stabilizers, binders, antioxidants, preservatives, lubricants, suspending agents, viscosity or flavoring agents and the like. In addition, they can also be formulated with other active ingredients to provide a broad spectrum of antibiotic activity.

The dose to be administered depends on a wide range of conditions under which the subject is treated and the weight of the recipient, the parenteral route being preferred for generalized infections and the oral route for intestinal infections. The daily dose consists of about 15 to about 600 mg of active ingredient per kg of body weight of the subject at one or more doses per day. The preferred daily dose is in the range of about 80 to 120 mg of active ingredient per kg of body weight. The preferred daily dose for the compound of sodium salt of 3-carbamoyloxymethyl-7-methoxy-7- (2-thienylacetamido / decephalosporic acid is in the range of about 80 to 120 mg of active ingredient per kg of body weight.

Formulations may be administered in several unit dosage forms, for example in solid or liquid oral dosage forms. Single dose formulations, liquid or solid, generally contain from about 15 mg to about 1500 mg by weight of the active ingredient based on the total total weight of the composition. A dosage amount of about 250 mg to 1000 mg is preferred. For parenteral administration, a single dose is usually the pure compound in a slightly acidified sterile solution or in the form of a soluble powder intended for solution. Typical formulations of specific products are described below.

141

The unit dosage form consists of mixing 120 mg of 3-carbamoyloxymethyl-7-methoxy-7- (O-α-amino-phenylacetamido / decephalosporic acid as the sodium salt with 20 mg of lactose and 5 mg of magnesium stearate and placing 145 mg of the mixture in gelatin capsule No. 3. Similarly, using more active ingredient and less lactose, other dosage forms can be filled into gelatin capsules No. 3. By mixing together more than 145 mg of the ingredients, they can also be prepared larger capsules and compressed tablets and pills. The following examples illustrate the invention.

Tablet containing 125 mg of 7-methoxy-7- (O-α-carboxy-phenylacetamido) -cephalosporic acid

For one tablet 7-methoxy-7- (O-a-aminophenylacetamide) -cephalosporanic acid 125 mg corn starch / United States pharmacopoeia / 6mg dicalcium phosphate 192mg lactose / United States pharmacopoeia / 190mg

The active ingredient was ground with dicalcium phosphate, lactose and about half an inch of starch. The mixture was then granulated with 15% corn starch 6 mg paste and coarse. It is dried at 45 ° C and again sieved through sieve No. 16. The remainder of the starch and magnesium stearate is added and the mixture is compressed into tablets of approximately 1.27 cm in diameter, 800 mg each.

Parenteral solution containing 500 mg of 3-carbamoyl-oxymethyl-7-methoxy-7- (2-thienylacetamido / decephalosporic acid

Ampoule 3-Carbamoyloxymethyl-7-methoxy-7- (2-thienylacetamido / decephalosporanoic acid sodium salt 500 mg

Dilute sterile water for injections 2 cc

Substitution of the equivalent amount of 7-methoxy-7- (α-carboxyphenylacetamido / cephalosporanoic acid with 500 mg of the 3carbamoyloxymethyl-7-methoxy-7- (2-thienylacetamido / decephalosporanoic acid) sodium salt, considered in the preceding example, also yields the form, corresponding to the above example. parenteral administration.

Ophthalmic solution containing 100 mg of 3-carbamoyloxymethyl-7-methoxy-7- (2-thienylacetamido / decephalosporanoic acid sodium salt 3-carbamoyloxy-7-methoxy-7- (2-thienylacetamido / decephalosporic acid sodium salt 100 mg of hydroxypropylmethyl) mg sterile water to 1 ml

Ear solution containing 100 mg. 3-Carbamoyloxymethyl] -7-methoxy-7- (2-thienylacetamido / decephalosporanoic acid sodium salt Sodium 3-carbamoyloxy-7-methoxy-7- (2-thienylacetamido / decephalosporanoic acid 100 mg benzalkonium chloride 0.1 mg sterile water 1 ml

Topical application containing 100 mg 3carbamoyloxymethyl-7-methoxy-7- (2-thienylacetamido / decephalosporanoic acid sodium salt 3-carbamoyloxymethyl-7-methoxy-7/2-thienylacetamido / decephalosporic acid sodium sodium salt 4000 mg 400 mg polyethylene glycol 400 US pharmacopoeia 1.0 g

142

The active ingredients of the above forms may be administered alone or in combination with other biologically active ingredients, for example with other antibacterial agents such as lencomycin, penicillin, streptomycin, novobiocin, gentamicin, neomycin, colistin and kanamycin, or with other therapeutic agents such as probenacid.

The new cephalosporins according to the invention can be used in the form of free acids, such as alkali metal, alkaline earth metal or ammonium salts, for example sodium, potassium, calcium, triethylammonium, procaine and the like, such as esters, especially labile esters such as acetoxymethyl or pyetoxymethyl and the like, or as amides.

7- (N-5'-amino-5'-carboxyvaleramide) -3-carbamoyloxymethyl-7-methoxy-3cephem-4-carboxylic acid and derivatives thereof having different substituents in the 3rd position of the general formula -CH ₂ A , as defined above, are described and claimed in the following US patents by Edward O. Stapby and Justo M. Mata:

Cep. No. 19496 per. March 13, 1970 Cep. No. 51319 per. June 30, 1970 Cep. No. 96594 per. December 9, 1970 and Cep. №115,779 per. February 16, 1971

These products are not within the scope of the present invention.

Claims (4)

Patent Claims An antibacterial composition comprising an antibacterial amount of 3- A -7methoxy-7-acylamido-3-cephem-4-carboxylic acid, wherein A is methyl, hydroxyethyl, chloromethyl, bromomethyl or fluoromethyl, mercaptomethyl, methoxymethyl ,, n-propoxymethyl, methylthiomethyl, acetoxymethyl, propionyloxymethyl, benzoyloxymethyl, / rchlorobenzoyl / oxymethyl, / p-methylbenzoyl / oxymethyl, pivaloyloxymethyl, / 1-adamantyl / carboxymethyl, bu5-carboxymethyl, carboxymethyloxymethyl, -ethylcarbamoyl / oxymethyl, (N- (2-chloroethyl) carr Hydroxycarbamoyl / oxymethyl, / Nfenilkarbamoil / oxymethyl, N-p-sul10 fofenilkarbamoil / oxymethyl, p-karboksimetilfenilkarbamoiloksimetil, methoxycarbonyloxymethyl, izobutanoiloksimetil, tsiklobutilkarboniloksimetil, karbamoiltiometil 15 / etoksitiokarbonil / thiomethyl, / npropoksitiokarbonil / thiomethyl, / tsiklopentanoksitiokarbonil / thiomethyl, N, N- diethylthiocarbamoylthiomethyl, N-methylpiperazine-1-thiocarbonylthio20 methyl /, N, N-dimethylpiperazine-1thiocarbonylthiomethyl, 2-furoylthiomethyl, isothiouromethyl, (5-methyl 1,2,2-thiadiazol-2-yl) -thiol-methyl-2-yl-methyl-2- loxime25 til, 1-methyl-1,2,3,4-tetrazolyl-5thiomethyl, tosyloxymethyl, sulfamoyloxymethyl, 1-naphthyloxymethyl, 2furylacetoxymethyl, cinamoyloxymethyl, p-hydroxycinamoyloxymethyl, 30 p-phenyloxymethyl, 30 p-sulfoxomethyl oxymethyl or pyridinmethyl and acylamide is of the formula ³⁵ n and -N-C / CH ₂ / n ZR in which η is 0-4, Ζ is oxygen or sulfur and R is benzyl, p-hydroxybenzyl, 440 amino-4-carboxybutyl, methyl, cyanmethyl, 2-pentenyl, n-amyl , n-heptyl, ethyl, 3- or 4-nitrobenzyl, phenethyl, β, β-diphenylethyl, methyldiphenylmethyl, triphenylmethyl, 2-methoxyphenyl, 2, 645 dimethoxyphenyl, 2, 4, 6-trimethoxyphenyl, 3,5-dimethyl-4 -isoxazolyl, 3-butyl-5-methyl-4-isoxazolyl, 5-methyl-3-phenyl-4-isoxazolyl, 3- (2-chlorophenyl) -5-methyl-4-isoxazolyl, 3- (2,6-dichlorophenyl) -5-methyl-4 isoxazolyl, D-4-amino-4-carboxybethyl, D-4-N-benzoylamino-4-carboxy-butyl, p-aminobenzyl, o-aminobenzyl, m- aminobenzyl, (3-pyridyl) -methyl, 2-ethoxy-1-naphthyl, 3-carboxy2-quinoxalinyl, 3- (2,6-dichlorophenyl-5- (2-furyl) -4-isoxazolyl, 3- 1 phenyl- 4-isoxazolyl, 5-methyl-3- (4-guanidinophenyl) -4-isoxazolyl, 4-guanidinylmethylphenyl, 4-guanidinmethylbenzyl, 4-guanidinbenzyl, 4-guanidinophenyl, 2,6-dimethoxy-4-guany-1-phenylphenyl, -carboxymethylbenzyl, p-carbamoylmethylbenzyl, m-fluorobenzyl, m-bromobenzyl, p-chlorobenzyl, p-methoxybenzyl, 1-naphthyl ethyl, 3-isothiazolylmethyl, 4-iso-2-thiazolylmethyl, 5-isothiazolylmethyl, 4- pyridylmethyl, 5-isoxazolylmethyl, 4-methoxy-5-isoxazolylmethyl, 4-methyl-5-isoxazolylmethyl, 1-imidazolylmethyl, 2-benzofuranylmethyl, 2- 2 indolylmethyl, 2-phenylvinyl, 2-phenylethynyl, 2- (5-nitrofuranyl / vinyl, phenyl, o- methoxyphenyl, o-chlorophenyl, o-phenylphenyl, p-aminomethylbenzyl, 1- (5-cyanotriazolyl) methyl, 3 difluoromethyl, dichloromethyl, dibromomethyl, 1- (3-methylimidazolyl) methyl, 2- or 3- (5-carboxymethylthienyl) methyl, 2- or 3- (4-carbamoylthienyl) methyl, 2- or 3- (5-methylthienyl) methyl, 2- or 3- (5-methoxythienyl) methyl, 2 or 3- (4) -chlorothienyl / methyl, 2- or 3 (sulfothienyl) methyl, 2- or 3- (5-carboxythienyl) -methyl, 3- (1,2,5-thiadiazolyl) methyl, 3- (4-methoxy-1,2) , 5thiadiazolyl), methyl, 2-furylmethyl, 2- (5-nitrofuryl) methyl, 3-furylmethyl, 2-thienylmethyl, 3-thienylmethyl and tetrazolylmethyl; and their pharmaceutically acceptable salts or carboxylic acid protecting derivatives thereof, wherein the derivative group is methyl, tert-butyl, trichloroethyl, allyl, propargyl, benzyl, diphenylmethyl, o-nitrobenzyl, 3,5 dinitrobenzyl, p-methoxybenzyl, ace0 toximethyl, pivaloyloxymethyl, phenacyl, trichloroethoxycarbonyl trimethylsilyl or tributyl tin, in pharmaceutically acceptable carriers. Composition according to claim 1, characterized in that cefem- 4-Carboxylic acid is 3-carbamoyloxymethyl-7-methoxy-7- (2-thienylacetamido) -3-cephem-4-carboxylic acid or salts thereof. Composition according to claim 2, characterized in that the salt is a sodium salt.