HU184631B - Process for preparing cef-3-em-4-carboxylic acid derivatives with antiaiotic activity - Google Patents

Abstract

Cephalosporin antibiotics of the general formula <IMAGE> (wherein R<a> and R<b>, which may be the same or different, each represent a C1-4 alkyl group or R<a> and R<b> together with the carbon atom to which they are attached form a C3-7 cycloalkylidene group; and R<4> represents hydrogen or a 3- or 4-carbamoyl group) exhibit broad spectrum antibiotic activity, the activity being unusually high against gram-negative organisms such as strains of Pseudomonas organisms. A particular antibiotic compound of formula (I) possessing excellent antibacterial activity against strains of Pseudomonas organisms, as well as other valuable therapeutic properties, is (6R,7R)-7-[(Z)-2- (2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oxyimino)acetamido]-3-(1- pyridiniummethyl)-ceph-3-em-4 carboxylate. The invention also includes the non-toxic salts and non-toxic metabolically labile esters of compounds of formula (I). Also described are compositions containing the antibiotics of the invention and processes for the preparation of such antibiotics.

Description

The present invention relates to a process for the preparation of valuable antibiotic cef-3-em-4-carboxylic acid derivatives.

Single-bonded cephalosporin compounds are referred to herein as "cefam" as used in the specification using the term "cefam" in J. Amer. Chem. Soc., 1962, 84, 3400.

Antibiotic cef-3-em-4-carboxylic acid derivatives are widely used in the treatment of diseases caused by pathogenic bacteria in human and veterinary medicine. These compounds are particularly useful in the treatment of bacterial diseases that are resistant to other antibiotics, such as penicillin compounds, and in the treatment of patients who are sensitive to penicillin. In many cases, there is a need for antibiotics of the cef-3-em-4-carboxylic acid type, which are active against both Gram-positive and Gram-negative microorganisms, and extensive research has been conducted to develop various types of cef-3-em-4-carboxylic acids.

British Patent No. 1,399,086 describes a novel antibiotic cef-3-em-4-carboxylic acid containing a 7/3 (a-etherated-oxyimino) -acylamido group, wherein the oxyimino group is in synchronous configuration. This class of antibiotic compounds is characterized by high antibacterial activity against a range of Gram-positive and Gram-negative organisms, which is associated with particularly high stability against?-Lactamases produced by various Gram-negative organisms.

The discovery of this class of compounds prompted researchers in this field to develop compounds that exhibit more favorable properties against certain classes of organisms, namely, Gram-negative organisms.

British Patent No. 1,496,757 discloses antibiotic cef-3-em-carboxylic acids having a 70-acylamido group of formula (A) wherein R ¹ and R ^B substituents may vary widely, e.g. alkyl groups can be either with the carbon atom to which they are attached to form a C 3 -C 7 cycloalkylidene group, while m and n are each 0 or 1, with the proviso that m and n are 0 or 1, the compound is a isomer or a and mixtures of anti-isomers containing at least 90% of the syn isomer. The 3-position of the cephem molecule may be unsubstituted or contain one of a wide variety of possible substituents. These compounds show particularly good activity against Gram-negative organisms.

British Patent No. 1,522,140 further discloses antibiotic cef-3-em-carboxylic acids of formula B wherein R ^{4 is} furyl or thienyl; R ² is C 1-4 alkyl, C 3-7 cycloalkyl. furylmethyl or thienylmethyl; and R ^{3 is} hydrogen or carbamoyl, carboxyl, carboxymethyl, sulfo or methyl; the compounds are syn isomers or mixtures of color and anti-isomers containing at least 90% of the syn isomer. These compounds show good activity against a wide range of Gram-positive and Gram-negative organisms. In addition, the compounds are also very stable against β-lactamases produced by various Gram-negative organisms and have good in vivo stability.

Other compounds of similar structure have been developed from these compounds in further experiments to produce antibiotics which exhibit broader spectrum of antibiotic activity and / or activity against gram-negative organisms. In addition to the changes in the 7α-acylamido groups of the above formula, specific residues were introduced at the 3-position of the cephem molecule. For example, the 852427 number Belgian patent describes antibiotic QEF-3-em-4-carboxylic acids which include, in U.S. 1,399.086 U.K. patent, and wherein the group R ¹ in the formula (A) above various organic groups ideszámítva the 2-amidotriazol-4-yl group may also be substituted and the oxygen atom in the oximino group will be attached to an aliphatic hydrocarbon group which is itself substituted, for example a carboxyl group. In such compounds, the substituent at the 3-position is an acyloxymethyl, hydroxymethyl, formyl or optionally substituted heterocyclic thiomethyl group.

Belgian Patent No. 836.813 further describes cef-3-em-4-carboxylic acids wherein R ¹ may be substituted in the above formula (A), e.g.

2-aminothiazol-4-yl and the oxyimino group is a hydroxyimino or a blocked hydroxyimino group such as methoxyimino. In such compounds, the 3-position of the cephem molecule is replaced by a methyl group which may itself be substituted by one of the large number of Nleflyl residues described therein, such as pyridinium, which may also be substituted, for example, by a carbamoyl group. The above-mentioned patent does not attribute antibiotic activity to such compounds, which are only referred to as intermediates in the preparation of the antibiotic compounds described there.

Belgian Patent 853,545 discloses antibiotic cef-3-em-4-carboxylic acids wherein the 70-acylamido side chain is primarily a 2- (2-aminothiazol-4-yl) -2- (syn) methoxyiminoacetamido and

The 3-position substituent may be one of the large number of substituents disclosed in the aforementioned Belgian Patent No. 836.813. Exemplary compounds described herein are those substituted in the 3-position with pyridinium methyl or 4-carbamoylpyridinium methyl.

It has now been found that cef-3-em-4-carboxylic acid, which contains a certain group defined below at the 7/3 position and a pyridinium methyl group at the 3 position, has a particularly beneficial effect (described in detail below). ) against several pathogenic microorganisms.

The present invention relates to (6R, 7R) -7 [(Z) -2- (2-aminothiazol-4-yl) -2- (2-carboxyprop-2-oxyimino) acetamido] - 3- (1-Pyridinium methyl) ceph-3-em-4-carboxylate for the preparation of the antibiotic cephalosporin compound and its non-toxic salts.

The compounds of the present invention are syn isomers. The syn-isomeric form is defined by the group of configuration (1) with respect to the carboxamido group. As used herein, the color configuration i is structurally characterized by the group of formula (2).

184,631

Since the compounds of the present invention are geometric isomers, it is understood that mixtures with the corresponding anti-isomer also exist.

Also included within the scope of the invention are solvates (in particular hydrates) of the compound of formula (I). Also included within the scope of the invention are salts of the compounds of formula (I).

The compound of formula (I) may also exist in tautomeric forms (e.g., with respect to the 2-aminothiazolinyl group) and such tautomeric forms, such as the 2-iminothiazolinyl form, are also within the scope of the invention. In addition, the compounds of formula (I) may exist in a variety of bonded forms, for example, in which the 4-carboxyl group has a proton, while the carboxyl group in the 7-side chain is deprived of its proton. cover.

The compound of formula (I) exhibits a broad spectrum of antibacterial activity. The effect on gram-negative organisms is unusually high. This great effect extends to some Gram-negative strains producing 0-lactamase. The compounds also show high stability against (3-lactamases) produced by a wide range of gram-negative organisms.

It has been found that the chemical strains produced by the process of the present invention exhibit unusually high activity against strains of Pseudomonas organisms, such as strains of Pseudomonas aeruginosa, and show high activity against a number of strains of Enterobacteriaceae (e.g., Escherichia coli, Klebsiella pneumoniae, Shigella sonnei, Enterobacter cloacae, Serratia marcescens strains, Providence varieties, Proteus mirabilis, and particularly indole-positive Proteus organisms such as Proteus vulgaris and Proteus morganii) and also show activity against Haemophilus influenza.

The antibiotic properties of the compounds of the present invention greatly exceed those of aminoglycosides, such as amikacin or gentamicin. This is particularly the case against various strains of Pseudomonas organisms which are insensitive to commercially available antibiotic compounds. Unlike aminoglycosides, antibiotic cephalosporin compounds have low toxicity in humans. The use of aminoglycosides in human medicine is limited and complicated by the high toxicity of these antibiotics. Thus, the antibiotic cephalosporin compounds of the present invention have incomparable advantages over aminoglycosides.

Non-toxic salt derivatives formed by the reaction of one or both carboxyl groups present in the compound of formula I include salts with inorganic bases such as alkali metal salts (e.g., sodium and potassium salts) and alkaline earth metal salts (e.g., calcium salts); salts with amino acids (e.g., lysine and arginine salts); salts with organic bases (e.g., procaine, fenfllethylbenzylamine, dibenzylethylenediamine, ethanolamine, diethanolamine and N-methylglucosamine salts). Other non-toxic salt derivatives are acid addition salts, such as those with hydrochloric acid, hydrogen bromide, sulfuric acid, nitric acid, phosphoric acid, formic acid and trifluoroacetic acid. Salts may also exist in the form of resinates, for example salts with a polystyrene resin or a cross-linked polystyrene divinylbenzene copolymer resin having amino or quaternary amino groups or sulfo groups, or salts with a carboxyl group resin such as a polyacrylic acid resin. Soluble salts of the compounds of formula (I) with bases (for example, alkali metal salts, such as the sodium salt) are useful in therapy because they are rapidly distributed throughout the body after administration. However, where insoluble salts of the compound of formula (I) are required for special applications such as storage, such salts may also be prepared in conventional manner, for example, with suitable organic amines.

These and other salt derivatives, such as salts with p-toluenesulfonic acid and methanesulfonic acid, can be used as intermediates in the preparation and / or purification of compounds of formula (I), for example in the methods described below.

The compound of formula (I), i.e., (6R, 7R) -7-l (Z) -2- (2-aminothiazol-4-yl) -2- (2-carboxyprop-2-oxyircino) -acetamido3-3 - (1-Pyridinium methyl) ceph-3-em4-carboxylate and its non-toxic salts (e.g., sodium salt) have excellent general antibiotic activity. It exhibits an outstanding effect against strains belonging to Pseudomonas organisms. The compound has excellent antibacterial properties, which do not harm human serum and, in addition, have low activity against large amounts of vaccine. The bactericidal activity of the compound is rapid, even at concentrations close to the lowest inhibitory concentration. The active ingredient is well distributed in the body of small rodents and provides useful therapeutic levels following subcutaneous administration. In large mammals, the compound produces high and sustained serum levels after intramuscular administration. The half-life in large mammals suggests the possibility of a relatively long half-life in humans, so that less frequent dosing may be required for less severe infections. Experimental infections in mice with Gram-negative bacteria have been successfully treated with this compound and have been shown to have particularly excellent protection against strains of Pseudomonas aeruginosa, an organism that is usually insensitive to antibiotics containing cephalosporin. This protection was compared with the result of treatment with a ninoglycoside such as amikacin. Acute toxicity experiments in mice with this compound yielded LD _S0 values greater than 1.0 g / kg. Nephrotoxicity was not observed in rats at doses of 2.0 g / kg.

Pharmaceutical compositions containing the compound of formula (I) as an active ingredient can be used in the treatment of diseases caused by pathogenic bacteria in both humans and animals. Such diseases include respiratory diseases and diseases of the urinary tract.

The antibiotic compound of formula (I) and its non-toxic salts are prepared by:

a) a compound of formula II wherein B is> S or> S · * 0 (a- or β-); also the β dashed bond at positions 2, 3 and 4 indicates that the compound is a cef-2-em or cef-3-em compound or a salt such as a mineral acid such as hydrochloric acid, a salt with hydrogen bromide, sulfuric acid, nitric acid or phosphoric acid or with an organic acid such as methanesulfonic acid or p-toluene

-3184 631 sulfonic acid salt, or an N-silyl derivative, or a corresponding compound having the 4-position -COOR ⁵ , R ^{5 is} hydrogen, or a carboxyl-protecting group such as an ester-forming aliphatic or araliphatic alcohol or an ester-forming residue of phenol, silanol or stannanol, wherein the alcohol, phenol, silanol or stannanol preferably has from 1 to 20 carbon atoms and has an associated anion A such as a halide such as chloride or bromide or a trifluoroacetate anion acid of the formula wherein R ⁶ a carboxyl blocking group, e.g., R ^s to a specified group; and R ^{7 is} acylated with a protected amino group or a corresponding acylating agent; obsession

b) a compound of formula IV wherein R ⁷ , B and the dashed line are as defined above; R ⁸ and R ^8a represents a hydrogen atom or a blocking group is a carboxyl group independently; and X is an unsubstituted residue of a nucleophilic group such as acetoxy or dichloroacetoxy or a halogen such as chlorine, bromine or iodine, or a salt is reacted with a pyridine of formula (V), followed if necessary and / or do any of the following in the appropriate order:

1) converting a Δ ² isomer to the desired Δ ³ isomer,

2) converting a compound wherein B is > S to > O into a compound wherein B is > S,

3) converting a carboxyl group into a non-toxic salt, and

4) removing the carboxyl-protecting group and / or the N-protecting groups.

In the above process variant (a), the starting material of formula (II) is preferably a compound wherein B is > S and the dashed line represents cef-3-em. A starting material which is particularly suitable for use in process a) is N- (7-aminoceph- S-em- S -yl-methyl) -pyridinium-6'-carboxylate dihydrochloride because of its high purity.

Acylating agents useful in the preparation of the compound of formula (I) are acid halides, in particular acid chlorides or bromides. Such acylating agents may be prepared by reacting an acid of formula (III) or a salt thereof with a halogenating agent such as phosphorus pentachloride, thionyl chloride or oxalyl chloride.

Acylation with acid halides may be carried out in an aqueous or non-aqueous reaction medium, generally at a temperature of from -50 ° C to + 50 ° C, preferably from -20 ° C to + 30 ° C, optionally in the presence of an acid scavenger. Suitable reaction media include aqueous ketones such as 4 aqueous acetone, esters such as ethyl acetate, halogenated hydrocarbons such as methylene chloride, amides such as dinreylacetamide, nitriles such as acetonitrile, or a mixture of two or more thereof. Suitable acid scavengers include tertiary amines (e.g., triethylamine or dimethylaniline), organic bases (e.g., calcium carbonate or sodium bicarbonate) and oxiranes such as lower 1,2-alkylene oxides (e.g., ethylene oxide or propylene oxide) that bind the free acylation reaction.

The acids of formula (III) themselves can also be used as acylating agents in the preparation of the compound of formula (I). Acylation with acids of formula (III) is preferably carried out in the presence of a condensing agent, for example a carbodiimide, preferably Ν, Ν'-dicyclohexylcarbodiimide 4-compound N-ethyl-N'-7-dimethylaminopropylcarbodiimide; a carbonyl compound such as carbonyldiimidazole; or an isoxazolium salt such as N-ethyl-5-phenylisoxazolium perchlorate. application.

The acylation may also be carried out with other amide-forming acid derivatives of formula (III), such as an active ester, a symmetric anhydride or a mixed anhydride, pivalic acid or a halogenate such as a lower alkyl halide. Mixed anhydrides may also be formed with phosphorous acids such as phosphoric acid or phosphoric acid, sulfuric acid, aliphatic or aromatic sulfonic acids (e.g. p-toluenesulfonic acid). An activated ester may conveniently be prepared in situ using, for example, 1-hydroxybenzotriazole in the presence of a condensing agent as defined above. Alternatively, the activated ester may be pre-formulated.

The acylation reactions are carried out with both the free acids and their amide-forming derivatives, preferably in an anhydrous reaction medium such as methylene chloride, tetrahydrofuran, dimethylformamide or acetonitrile.

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

The acids of formula III and the corresponding acylating agents may also be prepared and used, if desired, in the form of their acid addition salts. For example, acid chlorides may conveniently be used as their hydrochloride salts and acid bromides as their hydrobromide salts.

The nucleophilic compound can be used to displace a wide variety of X substituents from the cephalosporin compound of formula (V). The ease of displacement is somewhat related to the pK _{a of} the HX acid from which the substituent is derived. Thus, atoms or groups derived from strong acids are generally easier to replace than atoms or groups derived from weak acids.

The displacement of the X group with pyridine can conveniently be accomplished by keeping the reaction partners in solution or suspension. The reaction is preferably carried out using 1-10 moles of the pyridine compound.

Nucleophilic displacement reactions may conveniently be carried out on compounds of formula IV wherein X is a halogen atom or an acyloxy group such as those discussed below.

Ciloxy groups

Compounds of formula (IV) wherein X is an acetoxy group are suitable starting materials for the nucleophilic displacement reaction, wherein the exchange is carried out with pyridine (V). Alternative starting materials in this class are compounds of formula IV wherein X is a residue of a substituted acetic acid such as chloroacetic acid, dichloroacetic acid and trifluoroacetic acid,

Substitution reactions on compounds of formula IV, when they have such X substituents, particularly when X is acetoxy, can be facilitated by the presence of iodide or thiocyanate ions in the reaction medium. These types of reactions are described in more detail in Nos. 1,132,621 and

184,631 in British Patent 1,171,603.

The X substituent may be derived from formic acid, a halogen formic acid such as chloroformic acid or a carbamic acid. 5

In the case where a compound of formula IV is used wherein X represents acetoxy or substituted acetoxy, it is generally desirable that R ^{8 in} formula IV be hydrogen and B is> S. In this case, the reaction 10 is preferably carried out in an aqueous medium, preferably at a pH of 5-8pH, in particular at a pH of 5.5-7.

The above process, using compounds of formula IV wherein X is a substituted acetic acid residue, may be carried out as described in British Patent No. 1,241,657.

In cases when using a compound (IV) of formula in which X is an acetoxy conveniently between csopo.l, the reaction is 30 ° C and 110 T are preferred, i.e. ²⁰ sen to 50 ° C and 80 ° C in the temperature range.

Halogen derivatives

Conveniently, compounds of formula (IV) wherein X is chloro, bromo or iodo can be used as starting materials for the nucleophilic displacement reaction with pyridine (V). When a compound of formula (IV) belonging to this class is used, B represents> S to O and R ⁸ represents a protecting group for a carboxyl group.

The reaction is conveniently carried out in a non-aqueous medium, preferably one or more organic solvents, preferably solvents of a polar nature such as ethers, for example dioxane or tetrahydrofuran, esters such as ethyl acetate, amides such as formamide and Ν, Ν-dimethylformamide, and ketones. Thus, acetone is included. In some cases, pyridine may be the solvent itself. Other suitable organic solvents are described in detail in British Patent No. 1,326,521. The reaction medium can be neither very acidic nor very basic. Reactions with compounds of formula IV wherein R @ 4 and R @ ⁸ are both carboxylic. ^45- blocking group, the 3-pyridinium methyl product is formed into the corresponding halogen salts, from which one or more of the salts can be obtained in one or more reactions; contains anion.

In the case of the compounds of formula IV wherein X is a halogen as described above, the reaction is conveniently carried out at a temperature of -10 ° C to + 50 ° C, preferably + 10 ° C to + 50 ° C. .

The isolation of the reaction product from the reaction mixture, which contains, for example, 55 unchanged cephalosporin starting materials and other materials, can be accomplished by various methods. Such methods include recrystallization, lonophoresis, column chromatography, and ion exchangers (e.g., chromatography on ion exchange resins) or the use of macroreticular resins.

The A ² -cephalosporin ester derivatives of the present invention can be converted to the corresponding Δ ³ by, for example, treating the Δ ^2- ester with a base such as pyridine or triethylamine. ⁶⁵

A cef-2-em reaction product may also be oxidized to the corresponding cef-3-em-1-oxide. The oxidation is carried out, for example, with a persaw, for example peracetic acid or m-chloroperbenzoic acid, and the resulting parent oxide can, if desired, be subsequently reduced as described below to give the corresponding cef-3-em sulfide.

When a compound is obtained in which B is > S > 0, it can be converted to the corresponding sulfide. The reduction of the corresponding acyloxysulfonium or alkyloxysulfonium salt, which is prepared in situ by reaction with, for example, acetyl oxide, in the case of the actoxysulfenium salt, can be accomplished in various ways. For the reduction, for example, sodium dithionite or iodide ions can be used, where the iodide ions are used in a potassium iodide solution and the reduction is carried out in a water miscible solvent such as acetic acid, acetone, tetrahydrofuran, dioxane, dimethylformamide or dimethylacetamide. The reaction may be carried out at a temperature in the range of -20 ° C to + 50 ° C.

Basic salts of the compound of formula (I) may be prepared by reacting an acid of formula (I) with a suitable base. For example, the sodium or potassium salts may be prepared using the 2-ethylhexanoate or hydrogen carbonate salt. The acid addition salts may be prepared by reacting a compound of formula (1) with an appropriate acid.

When the compound of formula (1) is obtained in the form of an isomeric mixture, the color isomer is isolated, for example, by conventional means such as crystallization or chromatography.

The compounds of formula (III) used as starting materials in the preparation of the compound of formula (I) according to the invention are used in the free acid, acid halides and corresponding acid anhydrides in the isomeric form or as a mixture of the isomers and the corresponding antiisomers. the syn isomer.

The acids of formula (III) may be prepared by reacting a compound of formula (VI) wherein R 'is as defined above and R ⁹ represents a carboxyl-protecting group, with a compound of formula (VII) wherein and R and T is halogen such as chlorine, bromine or iodine; sulfate or sulfonate, such as tosylate, is etherified and, after e:, the carboxyl protecting group R ^{9 is} removed. Separation of the isomers may be carried out before or after etherification. The etherification is generally carried out in the presence of a base such as potassium carbonate or sodium hydride, and preferably in an organic solvent such as dimethylsulfoxide, a cyclic ether such as tetrahydrofuran or dioxane, or an N, N-disubstituted amide such as dimethylformamide. Under these conditions, the oxyimino configuration remains substantially unchanged during the etherification reaction. The reaction may be quenched in the presence of a base by using the acid addition salt of a compound of formula VI. The base is used in an amount sufficient to rapidly neutralize the acid in question.

The acids of formula (III) may also be prepared by reacting a compound of formula (VIII), wherein R ⁷ and R ⁵ are as defined above, with a compound of formula (IX), wherein R (R) is as defined above, carboxyl group 5

-5184 631 protecting the R ⁹ group and, if necessary, separating the colored anti-isomers,

The acids of formula (III) may be converted into the corresponding acid halides, acid anhydrides and acid addition salts by conventional means, for example as described above.

In the case where X is a halogen atom in the formula (IV), for example chlorine, bromine or iodine, the cef3-em starting compounds are conventionally used, for example, in the form of an O-protected amino-3-methylceph-3-err. Halogenation of 4-carbonic acid ester 1/5-oxide, removal of the 7 / J protecting group, acylation of the resulting 7/3-amino compound to give the desired β-acylamido group, for example analogously to process a) followed by the final step of reducing the Ιβ-oxide group. This method of preparation is described in British Patent No. 1,326,531.

The corresponding cef-2-em compounds can be prepared as described in Dutch Patent Application 6,902,013 by reacting a 3-methylcef-2-em compound with N-bromosuccinimide to give the corresponding 3-bromomethylcef-2-em compound. .

The starting materials of formula IV wherein X is an acetoxy group may be prepared, for example, by acylation of 7-amino-3-acetoxymethyl-cef-3-em-4-carboxylic acid as described in process a). Compounds of formula (V) wherein X represents a group other than acetoxy may be prepared by acylation of the corresponding 3-hydroxymethyl compounds, which may be prepared by hydrolysis of the corresponding 3-acetoxymethyl compounds, such as 1,474,519 and 1,531,212. United Kingdom Patents.

The starting materials of formula (II) may also be prepared in conventional manner, for example by nucleophilic displacement of the corresponding 3-acetoxymethyl compound 35 using the appropriate nucleophile. Such a method is described in British Patent No. 1,002,563.

Another method of preparing the starting materials of formula (II) is by deprotecting a suitable protected 7β-amino compound by conventional means such as PCl 2.

Some of the above transformations may require protection of some sensitive groups in the molecule of the compound in question from unwanted side reactions. For example, some of the above reaction processes may require protection of the NH ₂ group of the aminothiazolyl moiety, for example by tritylation, acylation (e.g., chloroacetylation), protonation 50, and other conventional methods. The protecting group can then be removed in a conventional manner without damaging the compound to be prepared. In the case of the trityl group, a halogenated carboxylic acid such as acetic acid, formic acid, chloroacetic acid or trifluoroacetic acid or a mineral acid such as hydrochloric acid or a mixture thereof may be used, preferably in the presence of a protic solvent such as water or chloroacetyl treatment with thiourea. 60

The carboxyl protecting groups used in the preparation of the compound of formula (I) or in the preparation of the required starting materials are required groups which are readily cleaved in the appropriate state of the reaction, preferably in the final step. 65

Suitable carboxyl blocking groups are well known in the art, one of which is disclosed in British Patent No. 1,399,086. Suitable carboxyl-blocking groups include aryl-lower alkoxycarbonyl groups such as p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl and diphenylmethoxycarbonyl; lower alkoxycarbonyl groups such as tert-butoxycarbonyl; and lower haloalkoxycarbonyl groups such as 2,2,2-trichlorotoxycarbonyl. Carboxyl-protecting groups can be removed, if necessary, by any suitable method described in the literature, such as acid or base-catalyzed hydrolysis, where appropriate, but also enzymatically-catalysed hydrolysis.

The antibiotic compounds of the present invention may be formulated for administration by any conventional route analogous to other antibiotic agents. The invention also relates to the preparation of pharmaceutical compositions containing the compound of formula (I) as active ingredients for use in human and veterinary medicine. Such formulations contain the necessary pharmaceutical carriers and excipients for normal use.

The antibiotic compounds of the present invention may also be formulated for injection, such as unit dosage forms in ampoules or multi-dose containers, for administration multiple times, with preservatives if necessary. The compositions may also be formulated as oily or aqueous suspensions, solutions, or emulsions, and may contain formulating agents such as suspending, stabilizing and / or dispersing agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, which may be reconstituted with sterilized, pyrogen-free water, before use.

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

Antibiotic compounds may also be formulated as suppositories containing conventional suppository bases such as cocoa butter or other glycerides.

For veterinary use, the compositions may also be formulated as intramammary compositions, both in long-acting and rapid-acting forms.

The compositions may contain more than 0.1%, e.g. 0.1-99%, of the active ingredient, depending on the route of administration. When the compositions are prepared in unitary form, they preferably contain from 50 to 1500 mg of active ingredient. In adult humans, the daily dose may range from 600 mg to 6000 mg daily, depending on the route and frequency of administration, for example, 1000-3000 mg daily for intravenous or intramuscular administration to adults. In the case of infection with Pseudomonas pathogens, higher doses should be administered daily.

-6184 631

The antibiotic compounds of the present invention may also be administered in combination with other therapeutic agents such as penicillin or other cef-3-em 4 carboxylic acid compounds.

The following examples illustrate the process of the present invention. Temperatures are given in degrees Celsius, petroleum ether solvent being a petroleum distillate in the range 40-60 °.

Proton magnetic resonance spectra (pmr) were determined at 100 MHz. The numbers correspond to the conventions, given in J, Hz; s = singlet, d = doublet, t = triplet, dd = doublet-doublet, in = multiplet, and ABq - AB quadruple.

The preparation of starting materials is illustrated in Examples A-E.

The example

Ethyl (Z) -2- (2-aminothiazcl-4-yl) -2- (hydroxyimino) acetate

To a solution of ethyl acetoacetate (292 g) in glacial acetic acid (296 ml) and ice-cooled was added a solution of sodium nitrite (180 g) in water (400 ml) at such a rate that the reaction temperature was maintained below 10 ° C. The mixture was further stirred and cooled for about 30 minutes, at which time a solution of 160 g of potassium chloride in 800 ml of water was added. The resulting mixture was stirred for one hour. The lower oil phase was separated and the aqueous phase was extracted with diethyl ether. The extract is combined with the oil, washed first with water and then with brine, dried and evaporated. The residual oil, which solidifies on standing, is washed with petroleum ether and dried in vacuo over potassium hydroxide. 309 g of ethyl (Z) -2- (hydroxyimino) -3-oxobutyrate are thus obtained.

To a solution of 150 g of ethyl (Z) -2- (hydroxyimino) -3-oxobutyrate in 400 ml of dichloromethane and ice-cooled is added 140 sulfuryl chloride dropwise. The resulting solution was kept at room temperature for 3 days and then concentrated. The residue was dissolved in diethyl ether and washed with water until almost neutral, then dried and evaporated. The residual oil, which was dissolved in ethanol (177 g), ethanol (500 ml), and dimethylaniline (77 ml) and thiourea (42 g) were added with stirring. After stirring for two hours, the product was isolated by filtration, washed with ethanol and dried to give 73 g of the title compound, m.p.

Example B

Ethyl (Z) -2-hydroxyimino-2- (2-tritylaminothiazol-4-yl) acetáthidroklorid

16.75 g of trityl chloride are added, with stirring over 2 hours, to 12.91 g of the product of Example A with 28 ml of dimethylformamide. containing 8.4 ml of triethylamine, prepared and cooled to -30. The mixture was allowed to warm to 15 ° over 1 hour, stirred for a further 2 hours, then partitioned between 500 mL of water and 500 mL of ethyl acetate. The organic layer was separated, washed with water (2 x 500 mL) and extracted with 500 mL of 1N HCl. The resulting precipitate was collected, washed with water (100 mL), ethyl acetate (200 mL) and ether (200 mL) and dried in vacuo. This gave 16.4 g of the title compound as a white solid, m.p. 184-186 °.

Example C

Ethyl (Z) -2- (2-tert-Butoxycarbonylprop-2-oxyimino} -2- (2-tritylaminothiazol-4-yl) acetate

Potassium carbonate (34.6 g) and tert-butyl 2-bromo-2-methylpropionate (24.5 g) in dimethylsulfoxide (25 mL) were added with stirring to a solution of the product of Example B (49.4 g) in dimethylsulfoxide (200 mL) for 6 hours. stirring at room temperature. The mixture is then poured into 2 liters of water, stirred for 10 minutes and given birth. The solid was washed with water and dissolved in 600 mL of ethyl acetate. The solution was washed successively with water, 2N hydrochloric acid, water and brine, and then concentrated. The residue was recrystallized from petroleum ether (b.p. 60-80 ° C) to give the title compound (34 g). Mp 123.5-125 °.

Example D (Z) -2- (2-tert-Butoxycarbonylprop-2-oxyimino) 2- (2-tritylaminothiazol-4-yl) acetic acid. The product prepared in Example C is dissolved in 20 ml of methanol and 3.3 ml of 2 Add sodium hydroxide solution. The mixture is refluxed for 1.5 hours and then concentrated. The residue was water (50 ml). It is taken up in a mixture of 7 ml of 2N hydrochloric acid and 50 ml of ethyl acetate. The organic layer was separated and the aqueous layer was extracted with ethyl acetate. The organic solutions were combined, washed with water, then with brine, dried and evaporated. The residue was recrystallized from a mixture of carbon tetrachloride and petroleum ether to give 1 g of the title compound, m.p. 152-156 °.

Example E (6R, 7R) -7-Amino-3- (1-pyridinium methyl) cef-3-em4-carboxylic acid dihydrochloride

a) A suspension of 4.15 g of (6R, 7R) -7- (2-thienylacetamido) -3- (1-pyridinium methyl) cef-3-em-4-carboxylate in 30 ml of dichloromethane with 5.09 ml of Ν, Ν-dimethylaniline and • treated with 2.52 ml of chlorotrimethylsilane. The mixture was stirred at 30-35 ° for one hour and then cooled to -28 and treated with 4.16 g of phosphorus pentachloride. Stir at -25 to -30 for an additional hour and then pour into a solution of 8.1 ml of butane-1,3-diol and 20 ml of dichloromethane cooled to -20 ° with stirring. The solution was allowed to warm to 0 ° over 30 minutes and the precipitated solid (A) was collected by filtration, washed with dichloromethane and dried in vacuo. The resulting material was dissolved in 17.5 mL of methanol, stirred and

Dilute with 87.5 ml dichloromethane. The solid precipitate was collected by filtration and washed with dichloromethane 7

-7184 631 and dried in vacuo to give 3.2 g of the title compound.

? max (pH 6-OS buffer) 258 nm (Ej 318);

τ (D ₂ O) values: 0.95, 1.32 and 1.84 (pyridinium protons), 4.10-4.46 (ABq, J = 16 Hz, 3-CH ₂ -), 4.56 (d, J = 5Hz, 7-H), 4.70 (d, J = 5Hz, 6-H), 6.14-6.50 (ABq, J = 17Hz, C ₂ -H).

b) 8 g of solid (A) prepared in step a) are dissolved in 25 ml of 1 N hydrochloric acid. To the solution is added 95 ml of isopropanol and the title product crystallizes as a dihydrate in an amount of 4.95 g.

τ (D ₂ O) values: 1.02, 1.36 and 1.87 (pyridinium protons); 4.2 + 4.55 (ABq, J = 14Hz, 3-CH ₂ -); 4.62 (d, J = 5 Hz, C 1 -H); 47.4 (d, J = 5 Hz, C ₆ -H); 6.19 + 6.38 (ABq, J = 18 Hz, C ₂ -H).

Water content as determined by Kari Fischer: 9.4%.

Example 1

a) Tert-butyl-1R, 7R) -3-acetoxymethyl-7 - [(Z) -2- (2-tert-butoxycarbonyl-prop-2-oximoyl) -2- (2-trityl-aminothiazol-4-yl) -acetamide] - ceph-3-em-4-carboxylate

A solution of 572 mg of the product prepared in Example D and 328 mg of tert-butyl (6R, 7R) -3-acetoxymethyl-7-aminoceph-3-em-4-carboxylate in 10 ml of dimethylformamide was cooled to 0 ° and 1 mg of 1-hydroxybenzotriazole followed by 225 mg of dicyclohexylcarbodiimide. The mixture was warmed to room temperature, stirred for 5 hours and allowed to stand overnight. The mixture was filtered and the white solid was washed with a little ether. The filtrate and washings were diluted with water (50 mL) and extracted with ethyl acetate. The organic extracts were combined, washed with water, 2N hydrochloric acid, water, sodium bicarbonate solution and brine, dried and evaporated. The residue was applied to a silica gel column and eluted with ether. The eluate containing product was collected and concentrated to give 533 mg of the title compound. Part of the product was recrystallized from diisopropyl ether, m.p. 103-113 ° (dec.).

[α] D 20 + 8.5 ° (c, 1.0, DMSO).

b) (6R, 7R) -3-Acetoxymethyl-7 - [(Z) -2- (2-aminothiazol-4-yl) -2- (2-carboxyprop-2-oxyimino (acetamido] cef3-em-4-carboxylic acid) ml Trifluoroacetic acid was added to a solution of the product obtained in 2.4 (ga) in 18 ml of anisole at 0 ° C. The mixture was stirred for 2 hours at room temperature and then concentrated. The residue was dissolved in ethyl acetate and extracted with saturated sodium bicarbonate solution. The aqueous phase was acidified to pH 1.5 under ethyl acetate, saturated with sodium chloride and extracted with ethyl acetate, and the combined organic extracts were washed with brine, dried and concentrated (50 mL). Dissolve in warm aqueous formic acid and allow to stand for 2 hours. The mixture is diluted with 50 ml of water and filtered, and the filtrate is concentrated. The residue is taken up in 50 ml of water, filtered again and lyophilized. 20 mg of the title compound are obtained.

Λ _max (pH 6 buffer) 236 nm (E 250);

255 nm (EJ *, 235), 296 nm (E '% _m 103);

[α] 25 D + 20.0 ° (c 1.0, DMSO).

c) (6R, 7R) -? - [(Z) -2- (2-Aminothiazol-4-yl) -2- (2-carboxy'-prop-2-oxyimino (acetamido] -3- (1-pyridiniummethyl)) cef-3-em-4-carboxylate monosodium salt ml of pyridine and 1.8 gb of the product obtained under (b) are added with stirring to a solution of 7.12 g of sodium iodide in 2.2 ml of water at 80 ° C. After stirring for 1 hour, cool and dilute to 100 ml with water, adjust the pH of the solution to 6.0 with 6.0 N sodium hydroxide solution, and concentrate the solution to remove pyridine. dilute to pH 1 with 2 drops of methyl isobutylchloride and acidify the solution to pH 1 with 2N hydrochloric acid, filter and wash the separated solids with a little water, combine the filtrate and washings with ethyl acetate, then adjust the pH to 6.0 with 2N sodium hydroxide solution, concentrate to 50 ml and add 500 g of Amberlite XAD-2 The column was loaded with water and then with 20% aqueous ethanol. The product-containing fractions were concentrated and lyophilized to give 0.56 g of the title product.

_.Lambda.max (pH 6 buffer) 253.5 nm (Ej307);

_{Λ max} 282 nm (ε f _m 159), 260 nm (E} +, 295);

[a] ² ? + 24.5 ° (c 1.0, DMSO).

Example 2 (6R, 7R) -7 - [(Z) -2- (2-Aminothiazol-4-yl) -2- (2-carboxyprop-2-oxyimino (acetamido] -3- (1-pyridinium methyl) cef) -3-em-4-karboxilátnátriumsó

2.5 g of (6R, 7R) -7- [2- (2-aminothiazol-4-yl) -2- (2-carboxyprop-2-oxyimino) acetamido] -3- (1-pyridiniummethyl) -ceph-3em4 Carboxylate was dissolved in water and treated with a solution of 1.52 g of sodium 2-ethylhexanoate in 8 ml of methanol.

The mixture was added with stirring to acetone over 15 minutes, the resulting suspension was filtered, the filtrate washed and dried to give the title compound (2.5 g).

[ ^α ] ²³ D (c 1.0, H ₂ O);

Xmax (pH 6 phosphate) 255 (Ej 327, e 18,630);

_Λnf 240 (Ejg, 305, e 17.370) and 280 (Ejg, 172, e 9-800);

rnax ^v (Nujol) 1780 ^cm-1 (β-lactam); Sodium Found: 4.5%, Calculated: C ₂₂ H ₂₁ O ₇ N ₆ S ₂ Na, 4.04%.

Example 3

o) Diphenylmethyl-η 5, 6R, N-dimethyl-carbonylprop-2-oxyimino) -2- (2-tritylammothiazol-4-yl) acetamido] -3-bromomethyl-ceph-3-em-N-oxide-4-carboxylate.

Phosphorus pentachloride (0.75 g) was suspended in methylene chloride (20 mL) with stirring. The mixture was cooled to -10 °

184,631 were cooled and 2.0 g of the product prepared in Example D was added. The mixture was stirred at -5 ° to -10 ° for 10 minutes, then 0.88 ml of triethylamine in 5 ml of methylene chloride was added at -10 °, followed by 1.67 g of diphenylmethyl (1S, 6R, 7R) after 5 minutes. A suspension of 7-amino-3-bromomethyl-ceph-3-imidoxide-4-carboxylate hydrobromide in 30 ml of methylene chloride containing 0.42 ml of triethylamine was added and washed with 5 ml of methylene chloride. The mixture was stirred for 20 minutes between -5 ° and -10 ° and then poured into 50 ml of a semi-saturated aqueous sodium bicarbonate solution. The organic layer was separated, washed with dilute hydrochloric acid (3X30 mL) and brine (2X30 mL) and concentrated in vacuo. The resulting foam was taken up in about 10 mL of ethyl acetate and treated with 100 mL of diisopropyl ether. The precipitate was collected by filtration, washed with diisopropyl ether and vacuum at 40 ° C overnight. dried to give 2.1 g of the title compound, r (CDCl ₃ ) values: 3.11 (s, -CH-Ph ₂ ), 3.37 (s, triazol-5-yl proton), 3.88 (dd, j = 9, 5 Hz, 7-H), 5.22 + 6.02 (ABq -3CH _2), 5.49 (d, 5Hz 6-H), 8.46 (s, CMe ₂ ).

b) (6R, 7R) -7 - [(Z) -2- (2-Aminothiazol-4-yl) -2- (2-carboxyprop-2-oxyimino) acetamido] -3- (1-pyridinium methyl) ceph-3-em-4-carboxylate

The product prepared according to Ig) is dissolved in 22 ml of acetone and the solution is stirred at room temperature. Pyridine (0.08 mL) was added and the mixture was stirred at room temperature for 3 hours. A further 0.72 ml of pyridine was then added and the mixture was allowed to stand at room temperature overnight. The mixture was poured into diethyl ether (75 mL) with stirring and the precipitated solid was collected by filtration, washed with ether and dried at 40 ° C under vacuum. The resulting solid (0.8 g) was redissolved at -10 ° in 22 mL of acetone and 0.7 g of potassium iodide was added followed by 0.17 mL of acetyl chloride. The mixture was stirred at -10 ° for 20 minutes, then 0.7 g of potassium iodide and 0.17 ml of acetyl chloride were added. The mixture was stirred for an additional 20 minutes at -10 ° and added to a solution of 0.6 g of sodium metabisulphite in 60 ml of water and 30 ml of saturated common salt. The product was extracted with methylene chloride (2 x 50 mL) and the extracts were washed with brine, dried over magnesium sulfate and concentrated under reduced pressure. The resulting foam

It is dissolved in 6.5 ml of formic acid and allowed to stand at room temperature for 15 minutes. Thereafter, 0.25 ml of concentrated hydrochloric acid solution was added and the mixture was allowed to stand for an additional 1.25 hours. The precipitated solid precipitate was collected by filtration and washed with a small amount of formic acid. The filtrate and wash were combined, poured into a mixture of ethyl acetate (5 mL) and diethyl ether (5 mL), and water (10 mL) and acetonitrile (5 mL) were added. More water is then added until two distinct layers are obtained. The bottom layer was run and extracted with 14 mL of diethyl ether containing 7 mL of Amberliter LAj and 0.7 mL of acetic acid. The aqueous layer was separated again and applied to a "Zeolite" 225 SRC 15 (H ⁺ form 15 mL) column. The column was washed neutral with water and the product eluted with 10% aqueous pyridine solution. The eluate was concentrated in vacuo to a small volume and treated with acetone. The mixture was cooled to 0-40 overnight and filtered. The precipitated solid was washed with acetone. and dried under vacuum at 40 ° to give 0.25 g of the title compound. The nmr spectrum is similar to that of Example 2.

λ _Γ18Χ (pH 6 phosphate) 255.5 nm (Ej 374),

_{Λ max} 238 (E340) and 290 nm (E140, 160).

Example 4

a) (6R, 7R) -7 - {(Z) -2- (2-triphenylmethylaminothiazol-4-yl) -2-t- (c-butoxika: bondprop-2-oxyimino) acetamido} 3-i -píridiniummetil) ceph-3-em-4-karboxiíit

44 g of the product obtained in Example D are added with stirring to a solution of 1.38 g of phosphorus pentachloride in 60 ml of methylene chloride and cooled to -10 °. The resulting solution was stirred at -5 ° for 30 minutes and then cooled to -Ιθ, followed by addition of 1.33 g of triethylamine followed by 20 ml of water. The mixture was stirred at 0 for 3 minutes, then the lower phase was added with stirring over 10 minutes to 2.19 g of 10a. Preparation of the product prepared according to Preparation II (30 ml) of Ν, Ν-dimethylacetamide and 30 ml of acetonitrile, containing 3.03 g of triethylamine, which is brought to -10 ° to -5 °. The mixture was stirred for 45 minutes at -10 ° to -5 ° for one hour without cooling and then 1 ml of methanol was added. Methylene chloride was removed by evaporation under reduced pressure and the remaining solution was poured into 300 ml of water with stirring to give 4.98 g of the title compound.

(CDCl 3) values: 2.78 (s, - [C ₆ H ₅ ] ₃ ); 3.37 L (s, thiazole proton); 0.35, 1.80, 2.12 pyridinium protons); 4.18 (m, -7-H); 4.95 (6-H); 8.66 (s-tert-butyl); 8.50 (s, -C (CH ₃ ) ₂ ).

b) (6R, 7R) -7 - [(Z) -2- (2-Amino-thiazol-4-yl) -2- (2-carboxyprop-2-oxymethyl) -acetate] -3- (pyridine); ) ceph-3-em-4-karbonsavdihidioklorid

3.38 g of the product of (a) are dissolved in 20 ml of 98% formic acid with stirring. Concentrated hydrochloric acid (1.2 ml) was added and the mixture was stirred for 1 hour. The precipitated solid was collected by vacuum filtration and the solvent was removed from the filtrate by evaporation under reduced pressure. An oily residue is obtained, which is triturated with 30 ml of acetone to give 2.20 g of the title product.

τ (D ₂ O / NaHCO ₃ ) values: 3.08 (s, -thiazole-propone); 1.06, 1.44, 1.93 (pyridine protons); 4.16 (d,

H-5 Hz, 7-H); 4.74 (d, J = 5 Hz, 6-H); 8.55 (s,

-C (CH ₃ ) ₂ ).

At acetone nmr, 1 mol.

Water content: 5% (Kari Fischer method).

Chlorine: found: 10.1% (C ₂₂ H ₂₄ N _e O ₇ S ₂ Cl ₂ + acetone (1 mol) + water (5%).

Chlorine: Calculated: 10.0%.

Example 5 j (6R, 7R) -7 - [(2} -2- (2-Riphenylmethylaminothiazol-4-yl) 2- (2-tert-butoxycarbonylprop-2-oximino) acetamido} 3- (1-pyridinium metal) ceph-3-em-4-carboxylate

2.18 g of the product prepared according to Example E (b) are now reacted according to the procedure of Example 4 to give

-9184 621

4.03 g of the title compound are obtained having spectroscopic properties similar to those of Example 4.

b) (6R, 7R) -7 - [(Z) -2- (2-Aminothiazol-4-yl) -2- (2-carboxyprop-2-oxyimino) acetamide] -3- (1-pyridiniummethyl) cef- 3-cephem-4-carboxylic acid dihydrochloride

3.8 g of the product of (a) were treated as described in Example 4 to give 2.17 g of the title compound having spectroscopic properties similar to those of Example 4.

Example 6

(a) (6R, 7R) -3-Acetoxymethyl-7 - [(Z) -2- (2-aminothiazol-4-yl) -2- (2-carboxyprop-2-oxyimino (-acetamido] -ceph-3em-4-) carboxylic acid hydrochloride

200 g, la. The product of Example 1d is dissolved in pre-cooled formic acid at + 10 ° C and concentrated hydrochloric acid (60 ml) is added with stirring over 5 minutes. Stirring was continued at 20-22 ° C for 1 1/4 hour, then cooled to + 10 ° C and filtered. The residue is washed with 30 ml of formic acid. The filtrate and wash were combined and concentrated at 20 *. This gave a yellow foam which was triturated with 800 mL of ethyl acetate. The precipitated solid was collected by filtration, washed with 200 mL of ethyl acetate and dried under vacuum at room temperature overnight to give 124.6 g of the title compound.

? _Max (ethanol) 234.5 nm,?

b) (6R, 7R) -7 - [(Z) -2- (2-Aminothiazol-4-yl) -2- (2-carboxyprop-2-oxyimido) acetamide] 3- (pyridinium-1-ylmethyl) cephem -3-em-4-carboxylate hydrate (ga) was added with stirring to a mixture of 40 ml of water and 25.6 ml of pyrine, followed by addition of 160 g of sodium iodide, which was heated at 60 ° for 3 1/2 hours. heat for a while. The warm solution was poured into 2 L of acetone with stirring and diluted with 1.2 L of diethyl ether. The slurry was cooled to 2 and the precipitate was collected by filtration to give 50.65 g of crude product, which was dissolved in 480 ml of water and 19.3 ml of formic acid and 280 ml of Amberlite LA2 in 560 ml of ether. The mixture was separated and the organic phase was washed twice with water (240 mL each). The aqueous layers were washed with ether (280 mL) and applied with a "Zeolite 225, SRC 15" (200 mL H ⁺ ) and then distilled with water until the eluate was neutral. The column was eluted with 10% aqueous pyridine and the eluate passed through a 40 g neutral alumina column. The eluate was then concentrated under reduced pressure to a syrup and the syrup was added dropwise to 500 ml of acetone with stirring to give 13.09 g of the cinch compound after filtration and equilibration in air.

H ₂ O content 7.0% (Kari Fischer method);

\ max 255 nm (E'§n 364),

The _NF 243 and 285 nm (Ei _m 338 and 171?), [ «Jd - 3 ° (pH 6 phosphate buffer).

Example 7

a) (6R, 7R) -7 - [(Z) -2- (2-Tritylaminothiazol-4-yl) -2- (2-tert-butoxycarbonylprop-2-oxyimino) acetamido] -5- (1-pyridiniummethyl) cef-3- em-4-carboxylate N, N-dimethylformamide solvate _r

Finely powdered'4á. The product of Example 1b is added at 23 ° C to 15 ml of N, N-dimethylformamide with stirring. The solid dissolves and shortly thereafter crystallization begins. The mixture was diluted with stirring by adding dropwise 20 ml of diisopropyl ether. The solid was collected by filtration to give 3.06 g of the title compound as needle crystals in the winter.

N, N-Dimethylfoamamide at nmr = 2 ¹ 1/2 mol. r (DMSO-d 6): 2.4-3.0 (m, trityl); 3.32 (s, aminothiazole ring proton); 0.47, 1.38, 1.82 (pyridinium protons); 4.34 (ni, c-7 proton); 4.92 (d, J-5, C-oh proton);

8.64 (s, tert-butyl protons); 8.62 (s, (CH _{3) 2} -CO, [ «Id - -27.5 ^° (c = 1.1 in methanol).

b) (6R, 7R) -7 - [(Z) -2- (2-Aminothiazol-4-yl) -2- (carboxyprop-2-oxyimino (-acetamido] -3- (1-pyridinium methyl)) cef- 3-cephem-4-karbonsavdihidroklorid

2.1 g of the product of (a) are dissolved in 10 ml of formic acid at 22 °. Concentrated hydrochloric acid (0.8 ml) was added and after 75 minutes the precipitate was collected by filtration. The filtrate was concentrated and industrial methylated alcohol (10 ml) was added. The solution was evaporated again, the residue was dissolved in methanol and the solution was added to diisopropylether to give 1.35 g of the title compound.

[α] D = -14.7 ° (c = 0.95), pH 6 buffer.

SHE

II τ (DMSO-d 6) 0.28 (d, J = 9, -C-NH), 0.77 (d, J = 6), 1.25 (t, J = 6), 1.70 (t , J = 6, pyridinium ring (40 protons); 3.0 (s, aminothiazole protons); 3.99 (dd, J = 9.5, 7-H); 4.67 (d, J = 5, 6-H); 8.42 (s, ~ (CH ₃ ) ₂ ).

pharmaceutical Example

Example A - Dry powder for injection

Ampoule (6R, 7R) -7-l (Z) -2- (2-aminothiazol-4-yl) -2- (carboxyprop-2-oxyimino) acetamido] -3- (1-pyridinium methyl) 50 cef- 3-em-4-carboxylate 500 mg lysine acetate 189 mg

Method

The antibiotic cephalosporin is mixed with Fzinacetate and filled into a glass vial. The vial head is flushed with nitrogen and sealed. The product is dissolved in 2 ml of water before administration.

⁶⁰

Example B - Dry powder for injection

Sterile (6R, 7R) -7-t (Z) -2- (Aminothiazol-4-yl) -2- (2-carboxyprop-2-oximino) acetamido] -3- (1-pyridiniummethyl) 65 eef-3- em-4-carboxylate monosodium salt in glass ampoules

-101

184,631 are filled so that each vial contains 1.0 g of antibiotic acid equivalent. Loading is performed under aseptic conditions under a sterile nitrogen blanket. To seal the vials, use a rubber disc or stopper fitted with an aluminum cap 5 to prevent gas exchange or the entry of microorganisms. The product is dissolved in water for injection before use, or other suitable sterile vehicle is used shortly before administration.

Example C - Twin Injection Kit

a) 500 mg of sterile (6R, 7R) -7 - [(Z) -2- (2-amino-thiazol-4-yl) -2- (2-carboxyprop-2-oxyloxy) acetanido] -3- (1-pyridine) amounts of methylmethyl) cef-3-em-4-carboxylate are filled into glass ampoules under a sterile nitrogen blanket. To seal the vials, we use a rubber disc or stopper fitted with an aluminum top closure to prevent gas exchange or the entry of microorganisms. 20

(b) A solution of 3.84% (v / v) sodium bicarbonate is prepared, purified by filtration and filled in 2.15 ml volumes into clean ampoules. Carbon dioxide is introduced into the vials one minute before sealing. The vials are autoclaved and checked for purity. 25

c) The antibiotic cephalosporin compound is dissolved shortly before administration in 2.0 ml of sodium bicarbonate solution.

Claims (4)

PATENT CLAIMS A process for the preparation of antibiotic (6R, 7R) -7 - [(Z) -2- (2-amino-thiazol-4-yl) -2- (2-carboxyprop-2-o; imino) -acetamide ] -3- (1-pyridinium methyl) ceph-3-em-4-carboxylate and its non-toxic salts, characterized in that: a) a compound of formula II wherein B is = S or = S- * O, while the dashed bond at positions 2, 3 and 4 indicates that the compound is a cef-2-em or cef-3-em compound, or a salt or N-silyl derivative thereof, or a 4-position -COOR ^S - wherein R ^s is hydrogen or a carboxyl group blocking group, preferably the residue of an ester-forming phenol, silanol or sztannanol - corresponding compound containing a group having an associated a ^- anion has formula (III) with an acid of formula - wherein R ⁶ a carboxyl group represented blocking group which may be preferably the same as R ⁵ substituents defined above, ^R7 is amino or protected, tritylated preferably acylated 10 or protonated amino group -, or an equivalent acylating or acylated with b) a compound of formula IV wherein R ⁷ , B and the dashed line have the same meaning as given in a); R ⁸ and R ⁸⁸ independently represent hydrogen or a blocking group is a carboxyl group, which is preferably the same as in a) ^and substituents defined in R; And X is an acyloxy group or a halogen atom - or a salt thereof - is reacted with a pyridine of the formula (V), and if necessary and / or desired, any of the following reactions are carried out in the appropriate order; 1) converting a Δ ² isomer to the desired Δ ³ isomer, 2) converting a compound wherein B is > S0 into a compound wherein B is > S, 3) converting a carboxyl group into a non-toxic salt, and 4) removing the carboxyl-protecting group and / or the N-protecting group. 2. Process for the preparation of process a) according to claim 1, characterized in that the starting material used is a cef-3-em compound of the formula II in which B is> S. A process according to claim 1 or a process according to claim 2, wherein the acylation of the compound of formula II is carried out with an acid halide corresponding to the acid of formula III. Process for process b) according to claim 1, characterized in that the starting material is a compound of the formula IV in which X is an acetoxy group or a bromine atom.