NO834304L - CEPHALOSPORINANTIBIOTIKA - Google Patents

Description

The present invention relates to cephalosporins. More specifically, it relates to new cephalosporin compounds and derivatives thereof with valuable antibiotic activity.

The cephalosporin compound in the present disclosure is named by reference to "cepham" after J. Amer. Chem. _ Soc., 1962, 84, 3400, the term "cephem" refers to the basic cepham structure with one double bond.

Cephalosporin antibiotics are widely used in the treatment of diseases caused by pathogenic bacteria in humans and animals, and are particularly useful in the treatment of diseases caused by bacteria that are resistant to other antibiotics such as penicillin compounds, and in the treatment of penicillin-sensitive patients. In many cases, it is desirable to use a cephalosporin antibiotic that shows activity against both gram-positive and gram-negative microorganisms, and a significant research effort has been devoted to the development of different types of broad-spectrum cephalosporin antibiotics.

In e.g. British patent no. 1,399,086 describes a new class of cephalosporin antibiotics containing a 73-(α-phorether oximino)-acylamido group, where the oximino group has the syn configuration. This class of antibiotic compounds is characterized by strong antibacterial activity against a number of gram-positive and gram-negative organisms associated with particularly high stability against 3-lactamases produced in various gram-negative organisms.

The discovery of this class of compounds has stimulated further research in the same area to find compounds with improved properties, e.g. against special classes of organisms, especially gram-negative organisms.

In British Patent No. 1,604. 971, a number of cephalosporin antibiotics are described in which the 7(3) side chain can be selected from, among others

_ a 2-(2-aminothiazol-4-yl)-2-(ether oxyimino) acetaminophen

A S V, II il

group, in which the ether group can be, among very many possible meanings, an alkyl group substituted with a cycloalkyl group, although there is no specific example of compounds with such a group. The 3-position group can also be chosen from a large number of alternatives and a possible substituent is a thiomethyl group substituted with an optionally substituted 5- or 6-membered heterocyclic group containing one or more nitrogen, oxygen or sulfur atoms.

British Patent Application No. 2027691A discloses cephalosporin antibiotics in which the 73-side chain is a 2-(2-aminothiazol-4-yl)-2-(ethered oxyimino)acetamido group, where the ethering group is a carboxyalkyl or carboxycycloalkyl group. The 3-substituent is a group of formula -CH2SY where Y is a carbon-bonded 5- or 6-membered heterocyclic ring containing at least one nitrogen atom, which may be substituted

with a C^_^alkyl group.

"It has now been found that by selecting a syn-2-(2-aminothiazol-4-yl)-2-cyclopropylmethoxyiminoacetamido group at the 70-position in combination with certain groups at the 3-position, cephalosporin compounds with particularly advantageous activity can be obtained (described in more detail below) against a number of common pathogenic organisms.

Accordingly, cephalosporin antibiotics of general formula (I) are prepared therein

where Y means a carbon-bonded 5- or 6-membered unsaturated heterocyclic ring containing at least one nitrogen atom, which ring may also contain one or more sulfur atoms and/or may be substituted with one or more C1_<j alkyl, oxo, hydroxy or carbaboylmethyl groups and non-toxic salts and non-toxic metabolically labile esters thereof. The compounds according to the invention are syn isomers. The syn-isomeric forms are defined by the configuration of the -group in relation to the carboxamido group. In the present description, the view configuration is structurally indicated as

It will be clear that because the compounds according to the invention are geometric isomers, some mixing with the corresponding anti-isomer may occur.

The invention also includes solvates (especially the hydrates)

of the compounds of formula (T). It also includes the solvates of non-toxic salts of formula (I) and non-toxic salts and solvates of non-toxic metabolically labile esters of the compound of formula (I). It should be clear that the solvates should be pharmacologically acceptable.

In formula (I) above, the heterocyclic ring represented by Y e.g. 1 to 4 nitrogen atoms and, if desired, a sulfur atom, particular examples of these heterocyclic groups are imidazolyl, pyrazolyl, pyridyl, pyrimidy 1, pyrazinyl, pyridazinyl, triazolyl, tetrazolyl, thiazolyl, thiadiazoly1, triazinyl and thiazolidiny1. The heterocyclic ring may, if desired, be substituted with one or more (e.g. one to three) C 1-4 alkyl (e.g. methyl), oxo, hydroxy or carbamoylmethyl groups. Alkyl or carbamoylmethyl substituents can e.g. is attached to the nitrogen heteroatom(s).

The compounds produced according to the invention show broad-spectrum antibiotic activity both in vitro and in vivo. They have good activity against both gram-positive and gram-negative organisms, including many 3-lactamase-producing strains. The compounds also have high stability against 3-lactamases produced by a number of gram-negative and gram-positive organisms.

Compounds according to the invention have been found to perform well

"activity against strains of Staphylococcus aureus and Staphylococcus epidermidis families including penicillinase-producing strains of these Gram-positive bacteria. "This is associated with good activity against various members of the Enterobacteriaceae (eg strains of Escher-ichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Serratia marcescens, Proteus mirabilis and indole-positive Proteus organisms such as Proteus vulgaris, Proteus morganii and Providence species), and strains of Haemophilus influenzae and Acinetobacter calcoaceticus as well as activity against some strains of Pseudomonas species. This combination of high activity against gram-positive organisms with high activity against gram-negative organisms in the prepared compounds is particularly unusual.

Non-toxic salt derivatives that can be formed by reaction

of the present carboxyl group in the compound of formula (I) or by reaction of any acidic hydroxyl group that may be present on the heterocyclic group Y includes inorganic base salts such as alkali metal salts (e.g. sodium and potassium salts) and alkaline earth metal salts (f .eg calcium salts); amino acid salts (e.g. lysine and arginine salts) and other organic base salts J

(eg procaine, phenylethylbenzylamine, dibenzylethylenediamine, ethanolamine, diethanolamine and N-methylglucosamine salts). Other non-toxic salt derivatives include acid addition salts, e.g. formed with hydrochloric acid, hydrogen bromide, sulfuric acid, nitric acid, phosphoric acid, formic acid and trifluoroacetic acid. The compounds can also exist in zwitter ion or internal salt form. The salts can also be present in the form of resins formed with e.g. a polystyrene resin or cross-linked polystyrene divinylbenzene copolymer resin containing amino or quaternary amino groups or sulfonic acid groups, or with a resin containing carboxyl groups, e.g. a polyacrylic acid resin. Soluble base salts (e.g. alkali metal salts such as sodium salt)

of the compound of formula (I) can be used for therapeutic applications due to the rapid distribution thereof

salts in the body after administration. When, however, insoluble salts of the compounds (I) are desired for a particular purpose, e.g. for use in depot preparations, can

such salts are formed in the usual way, e.g. with appropriate

—organic amines.

The group Y can have a positive charge, e.g. when Y is a 1-methylpyridinium-2-yl group, and when this is the case,

the positive charge must be balanced by a negative charge. Thus, the compounds can be (3-ines, so that the negative charge is provided by a -C00 Q group in the 4-position. Alternatively, the negative charge can be provided by an anion A 9 , so that an anion will be non-toxic and

can be derived from any of the above described acids which will form non-toxic salt derivatives.

Non-toxic metabolically labile ester derivatives which can be formed by esterification of the carboxyl group in the parent compound of formula (I) include acyloxyalkyl esters, e.g. lower alkanoyloxymethyl or -ethyl esters such as acetoxymethyl or -ethyl or pivaloyloxymethyl esters,

and alkoxycarbonyloxyalkyl esters, e.g. lower alkoxycarbonyloxyethyl esters such as the ethoxycarbonyloxyethyl ester. In addition to the above-mentioned ester derivatives include]

present invention the compounds of formula (I) in the form of other physiologically acceptable equivalents, i.e. physiologically acceptable compounds which, like metabolically labile esters, are transferred in vivo in the antibiotic parent compound of formula (I)

These and other salts and ester derivatives, such as the salts with toluene-p-sulfonic acid and methanesulfonic acid or the esters with t-butyl or diphenylmethyl esterification groups, can be used as intermediates in the preparation and/or purification of the present compounds of formula (I), e.g. in the methods described below.

Preferred compounds according to the invention include those where Y represents a 1-methylpyridinium-2-yl, ■

1- methylpyridinium-4-y1, 1-methyltetrazol-5-yl, 1-methyl-pyridinium-3-yl, 1, 2-dimethylpyrazolium-3-y 1, 1, 3-dimethyl-imidazolium-3-yl , 1-methylpyrimidinium-2-y1, 1-carbamoyl-methylpyridinium-4-y1, 2,5-dihydro-6-hydroxy-2-methyl-5-"oxo-1,2,4-triazin-3-yl, or 4,5-dihydro-6-hydroxy-4-methyl-5-oxp-1,2,4-triazin-3-yl group. A particularly preferred compound produced according to the invention is

(6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-cyclopropylmethoxyiminoacetamido]-3-[(1-methylpyridinium-4-yl)thiomethyl]-ceph-3 -em-4-carboxylate and its non-toxic salts.

The compounds produced according to the present invention may exist in tautomeric forms (e.g. with respect to

the 2-aminothiazolyl group or with respect to a 3-substituent such as a 2,5-dihydro-6-hydroxy-2-methyl-5-oxo-1,2,4-triazin-3-yl group), and it will be clear that such tautomeric forms fall within the scope of the invention.

The compounds according to the invention can be used to treat a number of diseases caused by pathogenic bacteria in humans and animals, such as respiratory tract infections and urinary tract infections.

L According to the invention, a method is provided by! preparation of cephalosporin compounds of the general formula

where Y is as above, B is or

(a- or 3~); the dotted line connecting the 2-, 3-, and 4-positions indicates that the compound is a ceph-2-em or ceph-3-em-f compound; R.^ means hydrogen or a carboxyl blocking group, e.g. the residue of an ester-forming aliphatic or araliphatic alcohol or an ester-forming phenol, silanol or stannanol (which alcohol, phenol, silanol or stannanol preferably contains 1 to 20 carbon atoms) or a symmetrical or mixed anhydride blocking group derived from a suitable acid; and R 2 denotes an amino or protected amino group, or salts thereof, characterized in that

(A) acylates a compound of the formula

where Y, B, the dotted line and r are as above,

or a salt, e.g. an acid addition salt (formed with e.g. a mineral acid^such as hydrochloric acid, hydrobromide, sulfuric acid, nitric acid or phosphoric acid or an organic acid such as methanesulfonic acid or toluene-4-sulfonic acid) or a 7-N-silyl derivative thereof, or alternatively

(when Y contains a quaternary nitrogen atom) a corresponding terminal compound with a group of formula -C00 in the 4-position, with an acid of formula

2

where R. is as stated above, or a salt thereof, or with an acylating agent corresponding thereto;

(B) reacts a compound of formula

12 where R , R , B and the dotted line are as indicated above, X is a substitutable residue of a nucleophile, e.g. an acetoxy or dichloroacetoxy group or a halogen atom such as chlorine, bromine or iodine, or a salt thereof, with a sulfur nucleophile serving to form the group -CH^SY (where Y is as above) in the 3-position; or (C) where Y in the compound of the invention contains a C 1-4 alkyl-substituted or carbamoylmethyl-substituted quaternary nitrogen atom in the heterocyclic ring, reacts a compound of formula

where R 2 , B and the dotted line are as indicated above;

R in this case is a carboxyl blocking group;

and Y' means a carbon-bonded 5- or 6-membered heterocyclic ring containing a tertiary nitrogen atom, with a C-^_4 alkylating agent or a carbanoylmethylating agent serving to introduce a C^_^ alkyl group or a carbamoylmethyl group as a substituent on the tertiary" nitrogen atom in the heterocyclic ring of Y'; after which, if necessary and/or desired in each case, one of the following reactions in appropriate order is carried out,

i) transfer of an A^-isomer into the desired A^-isomer,

ii) reduction of a compound where B is

while forming a compound where B is

iii) transfer of a carboxyl group in a non-toxic

metabolically labile ester function,

iv) formation of a non-toxic salt, and

v) removal of all carboxyl-blocking and/or N-protecting groups.

In the method (A) described above, the starting materials are

with formula (II) preferably a compound where B is

and the dotted line means a ceph-3-em compound.

When the group Y in formula (II) is charged, e.g. as in an N-alkylpyridinium group, and the compound contains a pit ! li of formula -COOR (where R is as indicated above) in the 4-position, the compound will include an associated anion E such as a halide, e.g. chloride or bromide, or trifluoroacetate anion.

Acylation reagents which can be used in the preparation of compounds of formula (I) include acid halides, especially acid chlorides or bromides. Such acylating agents can be prepared by reacting an acid (III) or a salt thereof with a halogenating reagent, e.g. phosphorus pentachloride, thionyl chloride or oxalyl chloride.

Acylations using acid halides can be carried out in aqueous or non-aqueous reaction media, preferably at temperatures from -50 to +50°C, preferably -40 to +30°C,

if desired in the presence of an acid binding reagent. Suitable

• reaction media include aqueous ketones such as aqueous acetone, aqueous alcohols such as aqueous ethanol, esters

such as ethyl acetate, halogenated hydrocarbons such as methylene chloride, amides such as dimethylacetamide, nitriles such as acetonitrile or mixtures of two or more such solvents. Suitable acid binding reagents include tertiary amines (eg, triethylamine or dimethylaniline), inorganic bases (eg, calcium carbonate or sodium bicarbonate), and oxiranes such as lower 1,2-alkylene oxides (eg, ethylene oxide or propylene oxide) which bind liberated hydrogen halide in the acylation reaction.

Acids of formula (III) can themselves be used as acylating agents in the preparation of compounds of formula (I). Acylations using acids (III) are preferably carried out in the presence of a condensing agent, e.g. a carbodiimide such as N,N<1->dicyclohexylcarbodiimide or N-ethyl-N'-γ-dimethylaminopropylcarbodiimide; a carboxyl compound such as carbonyldiimidazole; or an isoxazolium salt such as N-ethyl-5-phenylisoxazolium perchlorate.

Acylation can also be carried out with other amine-forming derivatives of acids with formula (III) such as e.g. an active right

I I

ester, a symmetrical anhydride or a mixed anhydride (eg formed with pivalic acid or with a haloformate such as a lower alkylhaloformate). Mixed anhydrides can also be formed with phosphoric acids (eg phosphoric acid or phosphoric acid), sulfuric acid or aliphatic or aromatic sulphonic acids (eg toluene-4-sulphonic acid). An activated ester

can easily be formed in situ by, e.g. using 1-hydroxy-benzotriazole in the presence of a condensing agent as described above. Alternatively, the activated ester can be formed beforehand.

Acylation reactions involving the free acids or their above-mentioned amide-forming derivatives are preferably carried out in an anhydrous reaction medium, e.g. methylene chloride, tetrahydrofuran, dimethylformamide or acetonitrile.

An alternative activation method is e.g. reacting an acid of formula (III) with a solution or previously formed suspension by adding a carbonyl halide, especially oxalyl chloride or phosgene, or a phosphoryl halide such as "phosphorus oxychloride" to a solvent such as a halogenated hydrocarbon, e.g. methylene chloride, containing a lower acyl tertiary amide such as N,N-dimethylformamide. The activated form of the acid of formula (III) can then be reacted with a 7-amino compound of formula (II) in a suitable solvent or mixture of solvents, e.g. a halogenated hydrocarbon e.g. dichloromethane. The acylation reaction can easily be carried out at temperatures from -50 to +50°C, preferably -40 to +30°C, if desired in the presence of an acid-binding reagent, e.g. as described above (eg, dimethylaniline).

If desired, the above-mentioned acylation reactions can be carried out in the presence of a catalyst such as 4-dimethylaminopyridine.

The acids of formula (III) and acylating agents corresponding to these can, if desired, be prepared and used in the form of

their acid addition salts. Thus, e.g. acid chlorides are easily used as the hydrochloride salts, and the acid bromides as in their hydrobromide salts.

In method (B) above, the sulfur nucleophile can be used to substitute a number of substituents X in the cephalosporin of formula (IV). To some extent, the ease of substitution depends on the pKa of the acid HX from which the substituent is derived. Thus, atoms or groups X derived from strong acids generally tend to be more easily substituted than atoms or groups derived from weaker acids. The ease of substitution is also to some extent related to the exact nature of the sulfur nucleophile. The latter nucleophile can e.g. is used in the form of a suitable thiol or thione.

The substitution of X with the sulfur nucleophile can be easily carried out by keeping the reactants in solution or suspension. The reaction is advantageously carried out using 1-10 molar equivalents of the nucleophile.

Nucleophilic substitution reactions can easily be carried out on such compounds of formula (IV) where the substituent X is a halogen atom or an acyloxy group, e.g. as discussed below.

Acyloxy groups

Compounds of formula (IV) where X is an acetoxy group are suitable starting materials for use in the nucleophilic substitution reaction with the sulfur nucleophile. Alternative starting materials within this class are compounds of formula (IV) where X is the residue of a substituted acetic acid, e.g. chloroacetic acid, dichloroacetic acid and trifluoroacetic acid.

Substitution reactions of the compounds (IV) with X substituents of this class, especially in the case where X is an acetoxy group, can be done more easily with iodide or thiocyanate ions present in the reaction medium.

The substituent X can also be derived from formic acid, a halogen-_ formic acid such as chloroformic acid, or a carbamic acid....J

When a compound of formula (IV) is used, wherein X means I

an acetoxy or substituted acetoxy group,. is it generally desirable that the group R"*" in formula (IV) should be a

hydrogen atom and that B is

In this case is performed

the reaction with advantage in an aqueous medium.

Under aqueous conditions, the pH of the reaction solution is advantageously kept in the range 6-8, if necessary by adding a base. The base is preferably an alkali metal or an alkaline earth metal hydroxide or bicarbonate such as sodium hydroxide or sodium bicarbonate.

When compounds of formula (IV) are used, where X is an acetoxy group, the reaction is preferably carried out at a temperature from 10° to 110°C, preferably 20° to 80°C.

Halogens

Compounds of formula (IV) where X is a chlorine, bromine or iodine atom can also be easily used as starting materials in the nucleophilic substitution reaction with the sulfur nucleophile.

When using compounds of formula (IV) in this class,

may B like to be

and R"*" can be a carboxyl blocker

group. The reaction is easily carried out in a non-aqueous medium which preferably comprises one or more organic solvents, with the advantage of polar nature such as ethers, e.g. dioxane or tetrahydrofuran, esters, e.g. ethyl acetate,

amides e.g. formamide and N,N-dimethylformamide, and ketones e.g. acetone. Other suitable organic solvents are described in more detail in British Patent No. 1,326,531. The reaction medium should be neither extremely acidic nor extremely basic.

When the reactions are carried out on compounds of formula (IV) where R is a carboxyl blocking group and the resulting group Y contains a quaternary close nitrogen atom,

the product will be formed as the corresponding halide salt, which, if desired, can be subjected to one or more ion-

l exchange reactions that give a salt with the desired anion.J

When using compounds of formula (IV) where X is a halogen atom as described above, the reaction is easily carried out at a temperature of -20° to +60°C, preferably 0° to +30°C.

When the incoming nucleophile does not give a compound containing a quaternized nitrogen atom, the reaction is generally carried out in the presence of an acid-binding agent, e.g. a base such as triethylamine or calcium carbonate.

In method (C) above, the compound of formula (V) is advantageously reacted with a compound of formula R 3Z

3

where R is a C^_^alkyl group or a group of formula r^NCOCP^-/ and Z is a leaving group such as a halogen atom (e.g. iodine, chlorine or bromine) or a hydrocarbonylsulfonate (e.g. mesylate or tosylate) group. Alternatively, a di-C^_^ alkyl sulfate, e.g. dimethyl sulfate, is used as an alkylating reagent. Iodomethane is preferred as the alkylating reagent and iodoacetamide is preferred as the carbamoyl methylation reagent. The reaction is preferably carried out at a temperature in the range from 0° to 60°C, preferably from 20° to 30°C. When the alkylating reagent is liquid under the reaction conditions as in the case of iodomethane, this reagent itself can serve as a solvent. Alternatively, the reaction can easily be carried out in an inert solvent such as an ether, e.g. tetrahydrofuran, an amide, e.g. dimethylformamide, a lower alkanol, e.g. ethanol, a lower di-alkyl ketone, e.g. acetone, a halogenated hydrocarbon, e.g. dichloromethane or an ester, e.g. ethyl acetate.

The compound with formula (V) used as starting material in method (C) can e.g. is prepared by reacting a compound of formula (IV) (as defined above) with a corresponding sulfur nucleophile in an analogous manner to the nucleophilic substitution reaction described in connection with method (B). If desired, the above-mentioned nucleophile can be used in the form of a metal thiolate salt.

When X in formula (IV) is halogen, the reaction for-j is carried out stepwise in the presence of an acid-binding agent, e.g. a base such as triethylamine or calcium carbonate.

The reaction product can be separated from the reaction mixture, which e.g. may contain unchanged cephalosporin starting material and other substances, by a variety of methods including crystallization, iontophoresis, column chromatography and the use of ion exchangers (eg, by chromatography on ion exchange resins) or macroreticular resins.

EtA<2->cephalosporin ester derivative produced According to the method according to the invention can be transferred into the corresponding desired A derivative by e.g. treatment of the A - ester with a base, such as pyridine or triethylamine.

When a compound is obtained where B is a

can this be transferred into the corresponding sulphide by e.g. reduction of the corresponding acyloxysulfonium or alkoxysulfonium salt prepared in situ by reaction with e.g. acetyl chloride in the case of an acetoxysulfonium salt, the reduction e.g. carried out with sodium dithionite or with iodide ion such as in a solution of potassium iodide in a water-miscible solvent, e.g. acetic acid, acetone, tetrahydrofuran, dioxane, dimethylformamide or dimethylacetamide. The reaction can be carried out at a temperature from -20° to +50°C. Metabolically labile ester derivatives of the compounds of formula (I) can be prepared by reacting a compound of formula (I) or a salt or protected derivative thereof with the corresponding esterification reagent such as an acyloxyalkyl halide or alkoxycarbonyloxyalkyl halide (e.g. iodide) readily in a inert organic solvent such as dimethylformamide or acetone, and when necessary all protecting groups are removed.

Basic salts of the compounds of formula (I) can be formed

by reacting an acid of formula (I) with a suitable base. Thus, e.g. sodium or potassium salts are prepared by using the respective 2-ethylhexanoate or hydrogen-^

carbonate salt. Acid addition salts can be prepared by

reacting a compound of formula (I) or a metabolically labile ester derivative thereof with the corresponding acid.

When a compound of formula (I) is obtained as a mixture

of isomers, the syn isomer can be isolated by e.g. common methods such as crystallization or chromatography.

For use as starting materials for the preparation of compounds of the general formula (I) according to the invention, compounds of the general formula (III) and the amide forming derivatives thereof such as acid halides and anhydrides corresponding to these in their syn-isomeric forms or in the form of mixtures of the syn isomer and the corresponding anti isomers containing at least 90% of the syn isomer.

Acids of formula (III) and their derivatives can be prepared by etherification of a compound of formula

R 2

(<VII>)

C.COOR<4>

II

N\

OH

2 4

where R is as previously defined and R means hydrogen or a carboxyl blocking group; or a salt thereof, by selective reaction with a compound of the general formula

where T is a halogen atom, such as a chlorine, bromine or iodine atom; a sulfate group; or a sulfonate group, such as p-toluenesulfonate; followed by the removal of any carboxyl blocking group R 4-. Separation of the isomers can be carried out either before or after such etherification. For-|

the etherification reaction is preferably carried out in the presence of a base, e.g. potassium carbonate or sodium hydride, and is preferably carried out in an organic solvent, e.g. dimethyl sulfoxide, a cyclic ether such as tetrahydrofuran or dioxane, or an N,N-disubstituted amide such as dimethylformamide. Under these conditions, the configuration of the oxyimino group is essentially unchanged by the etherification reaction. The reaction should be carried out in the presence of a base if an acid addition salt of a compound of formula (VII) is used. The base should be used in sufficient quantity to quickly neutralize the acid present.

Acids of formula (III) can also be produced by reaction

of a compound of formula (IX)

2 4 "where R and R are as previously defined; with a compound of formula (X)

after which any carboxyl blocking group R 4 is removed,

and, if necessary, the syn and anti isomers are separated.

The acids of formula (III) can be converted into the corresponding acid halides and anhydrides and acid addition salts by usual methods, e.g. as previously described.

When X is a halogen (ie chlorine, bromine or iodine) atom in formula (IV), starting ceph-3-em compounds can be prepared in a conventional manner, e.g. by halogenation of a 73-protected amino-3-methylceph-3-em-4-carboxylic acid ester 1(3-oxide, removal of the 73-protecting group, acylation of the

resulting 73-amino compound while forming it

{ desired 73-acylamido group, e.g. in an analogous manner to forward-I L l procedure (A) above, followed by reduction of 13-ok1'syd_-'

the group later in the sequence. This is described in British '

patent No. 1,326,531. The corresponding ceph-2-em compounds can be prepared by the method from Dutch patent application no. 6,902,013 by reacting a 3-methylceph-2-em compound with N-bromosuccinimide to form the corresponding 3-bromomethylceph-2-em -connection.

When X in formula (IV) is an acetoxy group, such starting materials can e.g. is produced by acylation of 7-aminocephalosporanic acid, e.g. in an analogous manner to method (A) above. Compounds of formula (IV) where X means other acyloxy groups can be prepared by acylation of the corresponding 3-hydroxymethyl compounds, which e.g. can be produced by hydrolysis of the corresponding 3-acetoxy-methyl compounds, e.g. as described in British Patent Nos. 1,474,519 and 1,531,212.

Compounds of formula (II) can also be prepared on

usual way, e.g. by nucleophilic substitution of a corresponding 3-acyloxymethyl or 3-halomethyl compound with the corresponding nucleophile, e.g. as described in British Patent Nos. 1,012,943, 1,241,657, 2,027,691A and 2,046,261A.

A further method for preparing the starting materials of formula (II) comprises exposing a corresponding 70-amino compound, e.g. in the usual way, e.g. when using PC1C.

It must be clear that in some of the above-mentioned transformations it may be necessary to protect any sensitive groups in the molecule in question in order to avoid unwanted side reactions. E.g. during the entire reaction sequence mentioned above, it may be necessary to protect the Nr^ group in the aminothiazolyl residue, e.g. by tritylation, acylation (eg chloroacylation or formylation), protonation or other common method, so that the protecting group is stable under the reaction conditions I. through one or more synthetic steps. The protecting group can then be removed by any conventional means which does not cause degradation of the desired compound, e.g. in the case of a trityl group using an optionally halogenated carboxylic acid, e.g. acetic acid, formic acid, chloroacetic acid or trifluoroacetic acid or the use of mineral acid, e.g. hydrochloric acid or mixtures of such acids, preferably in the presence of a protic solvent such as water or in the case of a chloroacetyl group, by treatment with thiourea.

Carboxyl-blocking groups used in the preparation of compounds of formula (I) or in the preparation of necessary starting materials are desirably groups that can be easily cleaved at a suitable step in the reaction sequence, preferably at the last step. However, in some cases it may be appropriate to use non-toxic metabolically labile carboxyl blocking groups such as acyloxymethyl or -ethyl groups (e.g. acetoxymethyl or -ethyl

or pivaloyloxymethyl) or alkoxycarbonyloxyalkyl groups (eg ethoxycarbonyloxyethyl) and retaining these in the final product to give an appropriate ester derivative of a compound of formula (I).

Suitable carboxyl blocking groups are well known and a number of representative blocked carboxyl groups are found in British Patent No. 1,399,086. Preferred blocked carboxyl groups include aryl-lower alkoxycarbonyl groups such as p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl and diphenylmethoxycarbonyl; lower alkoxycarbonyl groups such as t-butoxycarbonyl; and lower haloalkyloxycarbonyl groups such as 2,2,2-trichloroethoxycarbonyl. The carboxyl blocking group can then be removed by any suitable method described in the literature, thus e.g. acid or base catalyzed hydrolysis is applicable in many cases, as is enzymatic catalyzed hydrolysis.

It would be clear that the use of amino-protecting and carboxyl-blocking groups is well known and relevant examples of such use are given in e.g. Theodora W. Greene, j "Protective Groups in Organic Synthesis" (Wiley-Interscience, New York, 1981), and J.F.W. McOmie, "Protective Groups in Organic Chemistry" (Plenum Press, London, 1973).

The antibiotic compounds produced according to the invention can be formulated for administration in any appropriate manner analogous to other antibiotics and the invention includes pharmaceutical mixtures containing an antibiotic compound produced according to the invention adapted for use in human or veterinary medicine. Such mixtures can be prepared for use in the usual way with the help of all necessary pharmaceutical carriers or excipients.

The antibiotic compounds according to the invention can be formulated for injection and can be presented in unit dose form, in ampoules, in multi-dose containers, if necessary with a ti.l-■"" added preservative. The mixtures can also take the form of suspensions, solutions or emulsions in oily or

aqueous carriers, and may contain formulation reagents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient can be in powder form for reconstitution with a suitable carrier, e.g. sterile, oxygen-free water, before use.

If desired, such powder formulations may contain a suitable non-toxic base to improve the water solubility of the active ingredient and/or ensure that the powder is reconstituted with water and the pH of the resulting aqueous formulation is physiologically acceptable. Alternatively, the base can be present in the water with which the powder is reconstituted. The base can e.g. be an inorganic base such as sodium carbonate, sodium bicarbonate or sodium acetate, or an organic base such as lysine or lysine acetate.

The antibiotic compounds can e.g. presented in a suitable form for absorption in the digestive tract, e.g. as

tablets or capsules.

IN

be antibiotic compounds can also e.g. be formulated]

IN

as suppositories, e.g. containing common suppository bases such as cocoa butter or other glycerides.

Mixtures for veterinary medicine can e.g. are formulated as internal breast preparations in either long-acting or quick-release bases.

The mixtures can contain from 0.1% and upwards, e.g. 0.1-99% of the active material depending on the administration method. When the mixtures comprise dosage units, each unit will preferably contain 100-3,000 mg, e.g. 200-2,000 mg of the active ingredient. The dosage used for treating humans will preferably be from 200 to 12,000 mg, e.g. 1,000-9,000 mg per day, depending on the route and frequency of administration. E.g. will in the treatment of adults 400 to 6,000 mg per day given intravenously or intramuscularly would normally be sufficient. When treating Pseudomonas infections, higher daily doses may be necessary. It must be clear that in some cases, for example when treating infants, smaller dosage units and daily doses may be desirable.

The antibiotic compounds according to the invention can be given in combination with other therapeutic agents, such as antibiotics, e.g. penicillins or other cephalosporins.

The following examples illustrate the invention. All temperatures are in °C. Sorbsil U30 is silica gel manufactured by Joseph Crosfield and Son in Warrington, Lancashire, England. THF is tetrahydrofuran. DMF is N,N-dimethylformamide. DMSO is dimethyl sulfoxide.

IN

Intermediate 1

Ethyl ( Z )- 2- cyclopropylmethoxyimino- 2-( 2- tritylaminothiazol-4- yl) acetate

Ethyl (Z)-2-hydroxyimino-2-(2-tritylaminothiazol-4-yl)-acetate, hydrochloride salt (30 g) was stirred with cyclopropyl methyl bromide (13.5 g) in DMSO (150 mL) containing potassium carbonate (30 g) under nitrogen at 21°C for 7 hours. The mixture was partitioned between methylene chloride and water. The aqueous layer was extracted with more methylene chloride and the combined organic solutions were washed with water. After drying with magnesium sulfate, the solution was concentrated and applied to a column of Sorbsil U30 (200 g). The column was eluted

with ethyl acetate (10 to 30%) in petroleum ether (boiling point 40-""60°C). Evaporation of the appropriate fractions gave the title compound (20.9 g); (ethanol) 324.5 nm (E^<%>403);

>-i o rn.a.x -i 0 limb

. _ 254.5 nm (E, 302), 259.5 nm (E, 267), 265 nm

in£ limb ' limb

-(e:° 229), 271.5 nm (E^ 190) and 294 nm (E, 111);

lem lem lem _, v (CHBr.,) 3398 (NH) , 1730 (ester) , and 1593 and 1491 cm max -j

(aromatic double bond).

Intermediate product 2

(Z)-2-cyclopropylmethoxyimino-2-(2-tritylaminothiazol-4-y1)-acetic acid.

The product from Intermediate 1 (20 g) was dissolved in ethanol

(200 mL) and sodium hydroxide (3.12 g) in water (40 mL) were added. The mixture was refluxed for 45 minutes during which precipitation took place. Some of the ethanol (about 150 ml) was distilled off and the rest was cooled. The mixture was partitioned between methylene chloride and water containing 2N hydrochloric acid (70 ml). The organic layer was washed with water,

and each aqueous layer was back-extracted with more methylene chloride. The combined organic layers were dried with magnesium sulfate and evaporated to give the title compound

1(20g) ; A. - (ethanol) 234.5 nm (E^% 383) 259.5 nm (Ej% I 3 mf. lem lem

242), 266 . 5 nm (e]% 226) and 272.5 nm (<E>^ 217); V

lem<J>_1lem max (Nujol) 3260 (NH) and 1685 cm (acid).

Intermediate product 3

Diphenylmethyl ( 6R , 7R )- 3- bromomethyl- 7-[( Z )- 2- cyclopropylmethoxyimino- 2-( 2- tritylaminothiazol-4- yl)- acetamido] ceph-3- em- 4- carboxylate

Oxalyl chloride (0.37 mL) was added to a solution of DMF (0.38 mL) in methylene chloride (10 mL) at -20°C under stirring and nitrogen. The mixture was stirred with ice water cooling for 10 minutes before cooling again to -20°C. (Z)-2-cyclopropylmethoxyimino-2-(2-tritylaminothiazol-4-yl)acetic acid (1.94 g) was added and the solution stirred with ice water cooling for 10 minutes before cooling again to -20°C. A suspension of di-"phenylmethyl (6R,7R)-7-amino-3-bromomethylceph-3-em-4-carboxylate hydrochloride salt (1.98 g) in methylene chloride (10 ml) containing N,N-dimethylaniline (1 .76 ml) was added A clear solution formed when the mixture was allowed to warm to

21°C within 1 hour. The solution was washed with dilute hydrochloric acid and water, and each wash was back-extracted with more methylene chloride and the combined extracts were dried with magnesium sulfate and evaporated to a small volume. This solution was filtered through Sorbsil U30 (50 g) in ethyl acetate and the eluate was evaporated. The residue (3.54 g) was crystallized from diethyl ether - petroleum ether (bp 40 to 60°C) to give the title compound (2.43 g) m.p. 135 to 147°C, [a]<21>-11.9° (c 0.6, CHCl 3 ).

Intermediate product 4

Diphenylmethyl ( IS, 6R, 7R )- 3- bromomethyl- 7-[( Z )- 2- cyclopropyl 1-methoxyimino- 2-( 2- tritylaminothiazol- 4- yl) acetamido] ceph-3- em- 4- carboxylate, 1 - oxide

Diphenylmethyl (6R,7R)-3-bromomethyl-7-[(Z)-2-cyclopropylmethoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]cephi-3-em-4-carboxylate (1.2 g) was dissolved in methylene chloride (10 ml) and 3-cloperbenzoic acid (292 mg) was added with stirring and ice water cooling. After 30 minutes, the solution was washed with aqueous sodium bicarbonate solution and water (2 times). After drying with magnesium sulfate, the solution was concentrated and purified on a column of Sorbsil U30 (50 g) eluting with ethyl acetate (50 to 80%) in petroleum ether (b.p. 40 to 60°C) to give the title compound

(1.1 g) ; [α]<21>+14.2° (c 1.23, CHCl 3 ); \ ntl 244 .5 nm {Elcm 26°)' 258.5 nm (E**m 232), 266 nm (E** '225 , and 272 nm (e|% 214).

limb

Example 1

(a) Diphenylmethyl ( 6R, 7R )- 7-[( Z )- 2- cyclopropylmethoxyimino-2-( 2- tritylaminothiazol-4- yl) acetamido]- 3-[( 1- methylpyridinium- 4- yl) thiomethyl] ceph - 3- em- 4- carboxylate, bromide

Diphenylmethyl (6R,7R)-3-bromomethyl-7-[(Z)-2-cyclopropylmethoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]ceph-3-em-4-carboxylate (1.2 g) was dissolved in THF (20 mL)

with stirring at 21°C and 1-methylpyrid-4-thione (162 mg) was added. After 2.25 hours and 3.75 hours, two additional portions of 1-methylpyrid-4-thione (50 mg each) were added. After 6 hours, the solution was diluted with diethyl ether (80 mL) and

the mixture was stirred at ice bath temperature for a few minutes. The precipitate was collected by filtration, washed with diethyl ether and dried to give the title compound (1.27 g),

[α] 21 -29.7° (c 0.71, DMSO), ^ (Nujol) 1784 (3-lactam),

UIUciX

1720 (ester) and 1675 and 1520 cm (amide).

(b) (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-cyclopropyl-methoxyiminoacetamido]-3-[. (1-methylpyridinium-4-yl)thiomethyl]-ceph-3-em-4-carboxylate, dihydrochloride salt

The above ester (1.1 g) was dissolved in formic acid (4 ml) and concentrated hydrochloric acid (0.29 ml) was added. After stirring at 21°C for 1.5 hours, the mixture was filtered and the filter cake was leached with formic acid. The combined filtrations<_>

I - I

was evaporated to a gum which was triturated with acetone and dried to give the title compound (560 mg), [ct]^1 -39.7°

(c 1.03, DMSO) , (<N>ujol) 3680 - 2200 (OH, NH and NH->^), 1780 (3-lactam), 1710 (COOH) and 1675 and 1550 (amide).

Example 2

(a) Diphenylmethyl ( 6R, 7R )- 7-[( Z )- 2- cyclopropylmethoxyimino-2-( 2- tritylaminothiazol-4- yl) acetamido]- 3-[( 1- methylpyridinium- 2- yl) thiomethyl] ceph - 3- em- 4-^ carboxylate

Diphenylmethyl (6R,7R)-3-bromomethyl-7-[(Z)-2-cyclopropylmethoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]ceph-3-em-4-carboxylate was stirred with 1-methylpyride -2-thion (162

mg) in THF (20 mL) at 21°C for two hours. Additional 1-methyl-pyrid-2-thione (160 mg) was added. After 4 days at 21°C, the solution was refluxed for 3 hours. Finely crushed calcium carbonate (excess) was added and the mixture was again stirred at 21°C for two days. The mixture was filtered and the filtrate was diluted with diethyl ether (80 mL). The precipitate was collected by filtration, washed with ether and dried to give the title compound (540 mg), ^j_n£(ethanol) 240.5 nm (e]% 245), 265 nm (e|% 208), 272 nm (e} % 198), 308 nm

limb limb limb

(Er* 91), n> (Nujol) 3700 - 2500 (NH), 1790 (3-lactam),

limb iricLx

1725 (ester), 1680 and 1520 cm (amide).

(b) (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-cyclopropyl-. methoxyiminoacetamido]- 3-[( 1- methylpyridinium- 2- y1) thiomethyl]- ceph- 3- em- 4- carboxylate, dihydrochloride salt

The above ester (480 mg) was dissolved in formic acid (2 ml) and concentrated hydrochloric acid (0.13 ml) was added. After stirring for 1 hour at 21°C, the mixture was filtered and the filter cake was leached with formic acid. The combined filtrates were evaporated and the residue triturated with acetone to give the title compound (300 mg) 0 (Nujol) 3700-2300 (NH^,

ITlclX _ 1

NH and OH), 1780 (3-lactam), 1710 (acid), and 1680 cm (amide). T(dg-DMSO) includes 0.90 (d, J6 Hz), 1.58 (t, J

\ 6Hz) , 1.88 (d, J 6Hz) and 2.12 (t, J 6 Hz) pyridyl protons;

3.09 (thiazole proton); and.8.88, 9.30 to 9.80, multiplets, cyclopropyl protons.

Example 3

(a) Diphenylmethyl ( 6R, 7R )- 7-[( Z )- 2- cyclopropylmethoxyimino-2-( 2- tritylaminothiazol-4- yl) acetamido]- 3-[( 1- methyl- 1- H-trazol- 5- y1) thiomethyl] ceph- 3- em- 4- carboxylate

Oxalyl chloride (0.37 mL) was added to a solution of DMF (0.38 mL) in methylene chloride (10 mL) with stirring at -20°C. The mixture was allowed to warm to 0°C for 5 minutes before re-cooling to -20°C. (Z)-2-cyclopropylmethoxyimino-2-(2-tritylaminothiazol-4-yl)acetic acid (1.94 g) was added and the solution was stirred at 0°C for 5 minutes. After further cooling

to -20°C, a suspension of diphenylmethyl (6R,7R)-7-amino-3[ (1-methyl-1-H-tetrazol-5-yl)thiomethyl] ceph-3-em-4-carboxylate (1.98 g) in methylene chloride (10 ml) containing dimethylaniline (1.26 ml) added. The solution was allowed to heat

to 21°C over 1 hour. The solution was diluted with more methylene chloride and washed with dilute hydrochloric acid. The aqueous layer was extracted with more methylene chloride and the combined organic layers were washed with water, dried over magnesium sulfate and evaporated to a small volume. The solution was chromatographed on Sorbsil U30 (70 g) in a gradient of ethyl acetate (10 to 50%) in petroleum ether (b.p. 40 to 60°C) to give the title compound (3.28 g) as a foam, [ α]<21>-83.4° (c 0.91, CHCl-J ,\) (CHBr ) 3400 (NH), 1780 (3-lactam), 1725 (ester) and 1685 and 1515 cm (amide).

(b) (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-cyclopropyl-methoxyiminoacetamido]-3-[(1-methyl-1-H-tetrazol-5 - y1) thiomethyl] ceph- 3- em- 4- carboxylic acid, trifluoroacetate salt

The above ester (3.14 g) was dissolved in anisole (6.5 ml) with stirring at 21°C and trifluoroacetic acid (25 ml) was

added. After 1 hour, water (1.5 ml) was added. After a further 5 minutes, the solution was evaporated to half the volume and diisopropyl ether (about 100 ml) was added).

The precipitate was collected by filtration, washed with diisopropyl ether and dried to give the title compound (2.02 g), ta]21 -40.7° (c 0.44, DMSO), ^ (Nujol) 3700 - 2200 (NH D — max , 3 OH, NH), 1770 (3-lactam), 1725 (acid) and 1690 cm (amide).

Example 4

a) Diphenylmethyl ( IS, 6R, 7R)- 7-[( Z)- 2- cyclopropylmethoxyimino- 2-( 2- tritylaminothiazol-4- y1) acetamido]- 3-[( 1- methyl1-pyridinium-4- yl) thiomethyl] ceph- 3- em- 4- carboxylate, 1- oxide, bromide

Diphenylmethyl (IS, 6R, 7R) -3-bromomethyl 1-.7- [(Z)-2-cyclopropyl-methoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]ceph-3-em-4-carboxylate , 1-oxide (700 mg) was stirred with N-methyl-pyrid-4-thione (125 mg) in THF (15 mL) at 21 °C for 3.5 h.

--Diethyl ether was added and the precipitate was collected by filtration, washed with ether and dried to give the title compound (700 mg), [a]21 -31.8° (c 0.75, CHC1-) , \>

Uo IU3.X -i (Nujol) 1795 (3-lactam), 1722 (ester) and 1675 and 1512 cm

(amide)

h ) Diphenylmethyl-(6R,7R)-7-[(Z)-(2-cyclopropylmethoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]-3-[(1-methylpyridinium-4-yl)thiomethyl] ceph:- 3- em- 4- carboxylate, mixed halogenides

Diphenylmethyl •( IS , 6R , 7R )-7-[(Z)-2-cyclopropylmethoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]-3-[(1-methylpyridinium-4-yl)thiomethyl]ceph -3-em-4-carboxylate, 1-oxide, bromide (0.30 g) was stirred with potassium iodide (0.24 g) in acetone (2 mL) and DMF (2 mL) under ice-water cooling. Acetyl chloride

(0.1 mL) was added and stirring and cooling continued

3 hours. The solution was poured into aqueous sodium metabisulfite solution and the precipitate was collected by filtration. The solid was washed with water and diethyl ether and dried to give the title compound (230 mg) with n.m.r., i.r. and HPLC

in accordance with the product from Example 1 (a).

in i

IN

Protecting groups can be removed as in Example 1(b), which gives (6R,7R)-7-[(Z)-2-(2-aminothiazol-4~yl)-2-cyclopropyl-methoxyiminoacetamido]-3-[(1 -methylpyridinium-4-yl)thiomethyl]- ceph-3-em-4-carboxylate, dihydrochloride salt.

Example 5

a) Diphenylmethyl ( IS, 6R, 7R )- 7-[( Z )- 2- cyclopropylmethoxyimino- 2-( 2- tritylaminothiazol-4- yl) acetamido]- 3-( pyrid- 4-ylthiomethyl) ceph- 3- em - 4- carboxylate, 1- oxide

Diphenylmethyl (IS,6R,7R)-3-bromomethyl-7-[(Z)-2-cyclopropyl-methoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]ceph-3-em-4-carboxylate, 1 -oxide (700 mg) was stirred with 4-mercapto-pyridine (115 mg) and finely ground calcium carbonate (700 mg) in acetone (20 mL) under reflux for 2 h 10 min. The mixture was cooled and filtered. The filtrate was concentrated and chromatographed on silica gel (50 g). The column was set up in methylene chloride and was eluted successively with methylene chloride, ethyl acetate,. and 10% ethanol in ethyl acetate. Evaporation of appropriate fractions gave the title compound

(260 mg) as a foam; [a]21 -14.9° (c 0.81, CHCl) >)

(CHBr-J 3485 (NH), 1808 (3-lactam), 1778 (ester) and 1681

-1

and 1512 cm (amide).

b) Diphenylmethyl ( IS, 6R, 7R )- 7-[( Z )- 2- cyclopropylmethoxyimino- 2-( 2- tritylaminothiazol-4- yl) acetamido]- 3-[( 1- methylpyridinium- 4- yl) thiomethyl] ceph- 3- em- 4- carboxylate, 1- oxide, iodide

Diphenylmethyl (IS,6R,7R)-7-[(Z)-2-cyclopropylmethoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]-3-(pyrid-4-ylthiomethyl)ceph-3-em-4 -carboxylate, 1-oxide (100 mg) was dissolved in methyl iodide (0.5 mL). After 2 hours at 21°C, the solution was diluted with diethyl ether and the precipitate collected by filtration. This was washed with ether and dried to give the title iodide (70 mg) which resembled the above mentioned bromide salt in n.m.r., i.r. and HPLC.

in !

The product can be reduced as in Example 4(b) and protecting groups removed as in Example 1(b), giving (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- cyclopropylmethoxyimino-acetamido]-3-[(1-methylpyridinium-4-yl)thiomethyl]ceph-3-em-4-carboxylate, dihydrochloride salt.

Example 6

a) (6R,7R)-7-[(Z)-2-cyclopropylmethoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]3-[(1,2,5,6-tetrahydro-2-methyl" -5, 6- dioxo-l, 2, 4- triazin-3- yl) thiomethyl] ceph- 3- em- 4- carboxylic acid

A solution of DMF (0.4 mL) in dichloromethane (0.5 mL) was treated at -10°C with oxalyl chloride (0.15 mL) and the mixture stirred for 15 minutes at -10°C. (Z)-2-cyclopropylmethoxyimino--• 2-(2-tritylaminothiazol-4-yl)acetic acid (721 mg) was added. This solution was stirred in an ice bath for 15 minutes. It was then added to a solution of (6R,7R)-7-amino-3-[(1,2,5,6-tetra-" ■hydro-2-methyl-5,6-dioxo-1,2, 4-triazin-3-yl)thiomethyl]-ceph-3-em-4-carboxylic acid (500 mg) (which was prepared as described in European Patent Application No. 0065748) in methanol denatured alcohol (6 ml), water (0 .65 mL) and triethylamine (-1.9 mL). The resulting solution was stirred at 0°C for 1 hour. After stirring for an additional 10 minutes at 21°C, the reaction mixture was partitioned between dichloromethane and 2N hydrochloric acid solution. The organic layer was separated, washed with water, dried and evaporated to recover the title acid as a foam (1.12 g); [α]Q-37.91° (c 0.91, DMSO),

v (CHBr.J 3700 - 2700 (OH), 3395 (NH), 1789 (3-lactam),

ma x j _ 1738 (ester + other carbonyl groups), 1669 and 1523 cm (amide).

b) (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-cyclopropyl-methoxyiminoacetamido]-3-[(1,2,5,6-tetrahydro-2 - methyl 1- 5, 6-dioxo- 1, 2, 4- triazin- 3- yl) thiomethyl] ceph- 3- em- 4- carboxylic acid, formate salt

The ester from step (a) (1.169 g) was dissolved in formic acid

(9.3 ml). The solution was stirred and water (3.2 mL) was added. The mixture was stirred at 21°C for 1.5 hours. The solid was then filtered off. The filtrate was concentrated to a small volume and diisopropyl ether was added. The resulting solid was collected by filtration, washed with diisopropyl ether and dried to recover the title compound as a solid (654 mg); [a]D-51.35° (c 0.74,

DMSO) , \> (Nujol) 3650 - 2700 (OH + NH), 1779 (3-lactam), ms x ,

1738 (ester + dioxo) 1675 + 1530 (amide) and 1625 (carboxylate).

Example A

Dry powder for injection

Fill sterile (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-cyclo-propylmethoxyiminoacetamido]-3-[(1-methylpyridinium-4-yl)-thiomethyl]ceph -3-em-4-carboxylate aseptically in glass bottles,

so that each glass bottle contains 1 g of alcohol-free, "pure" material. Flush the bottle necks with sterile nitrogen; close the bottles using rubber gaskets or plugs and metal seals (applied by shrinking). The product can be constituted by dissolving in water for injections or other sterile vehicles shortly before use.

Claims (5)

1. Procedure for the preparation of cephalosporin compounds of the general formula where Y means a carbon-linked 5- or 6-membered unsaturated heterocyclic ring containing at least one nitrogen atom, which ring may also contain one or more sulfur atoms and/or may be substituted with one or more C-[__4 alkyl, oxo, hydroxy or carbamoylmethyl groups; B is S or S-0; the dotted line between 2-, 3- and 4- the positions indicate that the compound is a ceph-2-em or ceph-3-em compound; R means a hydrogen atom or a carboxyl blocking group; and R<2> means an amino or protected amino group; or salts thereof, characterized in that (A) acylates a compound with the formula where Y, B, the dotted line and R1 are as above defined; or a salt or 7-N-silyl derivative thereof or a corresponding compound having a group of the formula -C00 in the 4--I' position, with an acid of the formula |I 2 where R is as above; or a salt thereof, or with an acylating agent corresponding thereto; (B) reacts a compound of formula where B, the dotted line, R 2 and R 1 are as defined above, and X is a substitutable residue of a nucleophile; or a salt thereof, with a sulfur nucleophile forming the group -CH2 SY (where Y is as above) in the 3-position; (C) where Y in the desired compound contains a C^_^ alkyl-substituted or carbamoyl-substituted quaternary nitrogen atom in the heterocyclic ring, reacts a compound with the formula where B, the dotted line and R 2 are as above, R"*" in this case is a carboxyl blocking group, and Y' means a carbon-linked 5- or 6-membered heterocyclic ring containing one tertiary nitrogen atom; with a C 1 -3 alkylating reagent or a carbamoylmethylation reagent which introduces a C 1 -3 alkyl group or a carbamoylmethyl group as a substituent on the tertiary nitrogen atom in the heterocyclic ring of Y'; or (D) (for the preparation of a compound where R<1> means a hydrogen atom and/or R 2 means an amino group) removes the carboxyl-blocking group from a compound of formula (Ia) where R is a carboxyl-blocking group and/or removes the amino-protecting group from a compound of formula (Ia) where R 2 is a protected amino group. 2. Method according to claim 1 for the production of cephalo- trace information compounds with the general formula where Y is as set forth in claim 1; or non-toxic salts or non-toxic metabolically labile esters thereof, characterized in that after steps (A), (B) or (C) one or more of the following reactions are carried out in any possible order: i) transfer of an A^-isomer into the desired A^-isomer, ii) reduction of a compound where B is under formation of a compound where B is i iii) transfer of a carboxyl group in a non-toxic| metabolically labile ester function, iv) formation of a non-toxic salt, and v) removal of any carboxyl end-blocking and/or N-protecting groups. 3. Process according to each of claims 1 and 2 in the preparation of a compound where Y means a 1-methylpyridinium-2-yl, 1-methyl-pyridinium-4-y1, 1-methyltetrazol-5-y1, 1-methylpyridinium-3 -y1, 1, 2-dimethylpyrazolium-3-.y 1, 1,3-dimethylimidazolium-3-yl, 1-methylpyrimidinium-2-y1, 1-carbamoylmethylpyridinium-4-yl, 2,5-dihydro-6-hydroxy -2-methyl-5-oxo-1,2,4-triazin-3-yl or 4,5-dihydrop-6-hydroxy-4-methyl-5-oxo-1,2,4-triazin-3-yl -group, characterized by the use of correspondingly substituted starting materials. 4. Process according to each of claims 1-3 in the preparation of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-cyclo- propy Ime took sy iminoacetami do] -3-[(1-methylpyridinium-4-yl)-thiomethyl] ceph-3-em-4-carboxylate. and non-toxic salts thereof, characterized by using correspondingly substituted starting materials. 5. Process for the preparation of a pharmaceutical mixture, which as active ingredient contains a compound of formula (I) as defined in claim 2, or a non-toxic salt or non-toxic metabolically labile ester thereof, characterized by mixing this compound or the salt or ester thereof with a pharmaceutical carrier or excipient.