JPS6127921A - Pharmacological composition containing cephalosporin compound as effective component - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] The present invention is directed to a cephalic acid with valuable antibiotic properties.

The present invention relates to a pharmaceutical composition for use in humans or animals as an antibiotic against bacterial infections, which contains a rosporin compound as an active ingredient.

The cephalosporin compounds herein are rJ, A
ler, Chllil, Soc, J Vol. 84, p. 3400 (1962), 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 resistant to other antibiotics such as penicillin compounds and in patients susceptible to penicillin. It is particularly useful in the treatment of In many cases, it is desirable to use cephalosporin antibiotics that are active against both Durham-positive and Durham-negative bacteria, and a significant amount of research has been directed toward the development of various types of broad-spectrum cephalosporin antibiotics. It is being

That is, for example, British Patent No. 1, 39 of the present inventors et al.
7β-(α
A new class of cephalosporin antibiotics containing -etherified oxyimino)-acylamido groups is described. Antibiotic compounds of this type are characterized by a combination of high antibacterial activity against a range of Gram-positive and Gram-negative microorganisms and a particularly high stability against β-lactamases produced by various Durham-negative bacteria. There is.

The discovery of compounds of this type has stimulated further research in the field, for example in an attempt to find compounds with improved properties against certain types of microorganisms, especially Gram-negative microorganisms.

British Patent No. 1,496.757 (
JP-A-50-105689) describes the formula [wherein R is a chenyl group or a furyl group, and RA and R8 can vary widely, for example, a C1-4 alkyl group or a group in which they are bonded. together with the carbon atom that is 0
a 7β-acylamido group-containing cephalosporin antibiotic, which can form a 3-7 cycloalkylidene group, and m and n are each 0 or 1 such that the sum of 1 and n is O or 1. The compounds have been described to be syn isomers or a mixture of syn and anti isomers containing at least 90% syn isomers. The 3-position of the cephalosporin molecule may be unsubstituted or contain one of a wide range of possible substituents.

These compounds were found to have particularly good activity against Gram-negative microorganisms.

Furthermore, British Patent No. 1,522,140 (Japanese Unexamined Patent Publication No. 51-91287) by the present inventors describes the formula [wherein R1 represents a furyl group or ζ represents a chenyl group, and R2 represents a C1
-04 alkyl group, Cs-07 cycloalkyl group, furylmethyl group or chenylmethyl group, and R
3 represents a hydrogen atom or a carbamoyl group, carboxy group, carboxymethyl group, sulfo group or methyl group)
A number of cephalosporin antibiotics have been described, which compounds are syn isomers or exist as syn and anti isomer mixtures containing at least 90% syn isomers. These compounds exhibit high antibacterial activity against a wide range of Gram-positive and Gram-negative microorganisms. These compounds also have good stability in vivo and high stability against β-lactamases produced by various Gram-negative microorganisms.

Other compounds of similar structure have been developed from these compounds in yet another attempt to find antibiotics with improved broad-spectrum antimicrobial activity and/or high activity against Gram-negative microorganisms. Such developments include changes in the 7β-acylamido group in the above formula as well as the introduction of a specific group in the 3-position of the cephalosporin molecule.
Cephalosporin antibiotic compounds within the general range of word order 4-44 = 6 = 4 No. 44 → are described and described, in which the group R in formula (A) above is 2-aminothiazole. It may be substituted by a variety of different organic groups, including the -4-yl group, and the oxygen atom in the oximino group is itself bonded to an aliphatic hydrocarbon group which may be substituted, for example, by carboxy. In such compounds, the substituent at the 3-position is an acyloxymethyl group, a hydroxymethyl group, a formyl group, or an optionally substituted heterocyclic thiomethyl group.

Furthermore, Belgian Patent No. 836,813 (Japanese Unexamined Patent Publication No. 51-75066) discloses that the group R in the above formula (A) may be substituted, for example, by a 2-aminothiazol-4-yl group, and Motodoro!
Cephalosporin compounds are described that have xiimino groups or blocked hydroxyimino groups, such as methoxyimino groups. In such compounds, the 3-position of the cephalosporin molecule is substituted by a pyridinium group, which may itself be substituted by any of the residues of a number of nucleophile compounds, such as those described in the above specification, such as by a carbamoyl group. is optionally substituted with a methyl group. According to said specification, such compounds have no antibacterial activity and these compounds are only mentioned as intermediates for the preparation of the antibiotics mentioned in said specification.

Belgian Patent No. 853,545
2-125188), the 7β-acylamide side chain is mainly 2-(2-aminothiazol-4-yl)-2
-(syn)-methoxyiminoacetamide group and the substituent at the 3-position is the Belgian Patent No. 836
, 813 @ Mei III, as broadly defined cephalosporin antibiotics are described. Compounds specifically exemplified in the specification include, for example, compounds substituted at the 3-position by a pyridinium methyl group or a 4-carbamoylpyridinium methyl group.

By appropriately selecting a few specific groups at the 7β-position in combination with either a pyridinium methyl group or a 3-carbamoylpyridinium methyl group or a 4-carbamoylpyridinium methyl group at the 3-position, the inventors have discovered a wide range of possibilities. Particularly advantageous activity against commonly encountered pathogenic microorganisms (detailed later)
It has been discovered that cephalosporin compounds having the following properties can be obtained.

The present invention is based on the general formula [wherein R and Rb may be the same or different and each is a C1-4 alkyl group (straight chain alkyl group, preferably methyl, ethyl, n-propyl or n-butyl, particularly methyl or or R and Rb together with the carbon atom to which they are attached form a C3-7 cycloalkylidene group, preferably a C3-5 cycloalkylidene group, and R4 is hydrogen or 3- carbamoyl group or 4-carbamoyl group] and their non-toxic salts and non-toxic metabolically unstable esters.

The compounds according to the invention are syn isomers. This syn-isomeric form is defined by the configuration of the group (2) with respect to the carboxamide group. In the present invention the thin configuration is shown as structural.

Since the compounds according to the invention are geometric isomers, it will be appreciated that some admixture with the corresponding anti-isomer may occur.

The present invention also includes within its scope solvates (especially hydrates) of the compounds of formula (I). It also includes within its scope salts and esters of the compounds of formula (I).

The compounds according to the invention may exist in tautomeric forms (e.g. with respect to the 2-aminothiazolyl group), and such tautomeric forms, e.g. the 2-iminothiazolinyl form, are included within the scope of the invention. Let's understand this. Furthermore, the compounds of formula (I) described above also exist in another zwitterionic form, for example in which the 4-carboxyl group is protonated and the carboxyl group in the 7-side chain is deprotonated. This alternative form is also included within the scope of the invention.

It will also be understood that when R and Rb in the above formula represent different C1-4 alkyl groups, the carbon atom to which they are bonded contains an asymmetric center.

Such compounds are diastereoisomers and the present invention encompasses individual diastereoisomers of these compounds as well as mixtures thereof.

The compounds according to the invention exhibit broad-spectrum antimicrobial activity.

Activity against Durham-negative microorganisms is unusually high. This high activity extends to many β-lactamase producing Gram-negative bacterial strains. These compounds also have high stability towards β-lactamases produced by a range of Durum negative microorganisms.

The compounds according to the invention are derived from Pseudomonas (P Seu-d
Microorganisms of the genus Pseudomonas, such as Pseudomonas aerugi
unusually high activity against strains of Enterobacteriaceae (e.g.
E 5cherichia coli, K 1ebsiella pn
eumoniae), Salmonella typhimurium (
S almonel la, typhimurium
), Shioella zonnei (Shioella 5one)
i) Enterobacter cloacani (Enterobacter cloacani)
obacter cloacae), Serratia marcescens,
Providence species, Proteus m1rabili
s) and especially for example Proteus vulgaris (Pr
Indole-positive Proteus microorganisms such as OteuS vulgaris and Proteus morganii) and Haemophilus influenzae.
It was found that it exhibits high activity against strains of S. uenzae).

The antibacterial properties of the compounds according to the invention compare very favorably with those of aminoglycosides such as, for example, amikacin or gentamicin. In particular, this applies to their activity against various Pseudomonas microorganisms that are not susceptible to many of the existing commercially available antibiotic compounds. Unlike aminoglycosides, cephalosporin antibiotics usually exhibit low toxicity in humans. The use of aminoglycosides in human treatment tends to be limited or complicated by the high toxicity of these antibiotics. That is, the cephalosporin antibiotics of the present invention have significant advantages over aminoglycosides.

Examples of non-toxic salt derivatives that can be produced by reaction of either or both of the carboxyl groups present in the compound of general formula (I>) include alkali metal salts (e.g. sodium and potassium salts) and alkaline earth metal salts. (e.g. calcium salts), amino acid salts (e.g. lysine salts and arginine salts), organic base salts (e.g. propidence, phenylethylbenzylamine salts, dibenzylethylenediamine salts, ethanolamine salts, jetanolamine salts and Examples of other non-toxic salt derivatives include hydrochloric acid, hydrobromic acid, sulfuric acid,
Mention may be made of acid addition salts formed with nitric, phosphoric, formic and trifluoroacetic acids. These salts may also be used, for example, in polystyrene resins containing amino or quaternary amino groups or sulfonic acid groups or in resins made from cross-linked polystyrene-divinylbenzene copolymer resins or carboxyl group-containing resins such as polyacrylic acid resins. It may be in the form of an acid salt. Soluble base salts (eg, alkali metal salts such as the sodium salt) of compounds of formula (I) may be used therapeutically because upon administration, such salts are rapidly distributed within the body. However, if insoluble salts of compound (I) are desired for use in a particular application, eg, in a devo formulation, such salts may be prepared in conventional manner, eg, using a suitable organic amine.

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

Examples of non-toxic metabolically unstable ester derivatives produced by esterifying one or both carboxyl groups in the raw compound of formula (I) include acyloxyalkyl esters, such as lower alkanoyloxymethyl; Mention may be made of esters or lower alkanoyloxyethyl esters such as acetoxymethyl ester or acetoxyethyl ester or pivaloyloxymethyl ester.

Besides the above-mentioned ester derivatives, the invention also includes within its scope other physiologically acceptable equivalent forms of the compounds of formula (I), i.e. ) that is converted into the original antibiotic compound.

More preferred compounds of the present invention due to their high antibacterial activity are compounds in which R4 represents hydrogen in the above formula (I), i.e. compounds of the general formula [wherein Ra and Rb have the above definitions] and their non-toxic properties. Salts and non-toxic metabolically unstable esters.

A notable compound of formula (Ia) is (6R,7R)-7-(<Z')-2-(2-
aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide)-3-(1
-pyridinium methyl)-cef-3-em-4-carboxylate and its non-toxic salts (e.g. sodium salt) and non-toxic metabolically unstable esters.

Compounds of formula (Ib') significantly possess the general antimicrobial properties mentioned above for compounds of general formula (I).

However, it emphasizes its excellent activity against strains of Pseudomonas microorganisms. This compound has excellent antibacterial properties that are not inhibited by human serum, and furthermore the effect of increased inoculum on the compound is low. This compound is rapidly bactericidal at concentrations near the minimum inhibitory concentration. After subcutaneous injection, it is sufficiently distributed throughout the body of a petrodont to provide useful therapeutic levels. In primates, this compound provides high and long-lasting serum levels after intramuscular injection. Serum half-lives in primates indicate the potential for relatively long half-lives in humans, and less severe infections require the possibility of less frequent dosing.

Experimental infections with Durham-negative bacteria in mice have been successfully treated using this compound, and in particular Pseudomonas phi aeruginosa, an organism not normally susceptible to treatment with cephalosporin antibiotics.
Significant protection was obtained against S. monasaeruginosa strains. This protection was comparable to treatment with aminoglycosides such as amikacin. Acute toxicity studies on this compound in mice gave L Dso values of greater than 1.09/Kg. No nephrotoxicity was observed in rats at a dose of 2.09/My.

Another compound with properties not significantly different from the compound of formula (Ib) is (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclobut -1-oximino)acetamido)-3-(1-pyridiniummethyl)-cefu 3-emu 4-carboxylate and its non-toxic salts and non-toxic metabolically unstable esters.

Other examples of preferred compounds according to the invention include, for example, the following compounds of formula (I) and their non-toxic salts and non-toxic metabolically unstable esters.

(6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2
-oximino)acetamide]-3-(4-carbamoyl-1-pyridiniummethyl)-cef-3-em-4
-carboxylate, <6R,7R>-7-[(Z)-
2-(2-aminothiazol-4-yl)-2-(1-
Carboxycycloprop-1-oxyimino)acetamide)-3-(1-pyridiniummethyl)-cef-3-
Em-4-carboxylate, (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclobent-1-yloxyimino)acetamide) -3-(1
-pyridinium methyl)-cef-3-em-4-carboxylate and <6R,7R)-7-[(Z)-2=
(2-aminothiazol-4-yl)-2-(1-carboxycyclobut-1-oximino)acetamide]-3-(4-carbamoyl-1-pyridiniummethyl)-cef-3-em-4-carboxylate .

Examples of other compounds according to the invention include, for example, formula (I)
Mention may be made of compounds in which the groups Ra, Rb and R4 are as follows.

Ra Rb R4 a) Zl main and 1 -CH3-02Hs H -02R5-C2Hs -CHs -CH33-CONH2C2R5 C2Hs -CHs 4-CONH2C2H5 Cyclobutylidene 3- C,ON H2cyclo0
Pentylidene 3-CONH24-CONH2 Schiff 0 hexylidene R 3-CONH2 4-CONH2 Cyclopropylidene 3-CONH24-CONH
2 The compounds of formula (I) can be used to treat various diseases caused by pathogenic bacteria in humans and animals, such as respiratory tract infections and urinary tract infections.

According to another aspect of the invention, formula (A) (wherein R4 has the above definition and B is 〉S or 〉S→
0 (α- or β-) and the dotted line bridging the 2-, 3-, and 4-positions indicates that the compound is a cef-2-em or cef-3-em compound. or a salt thereof, such as an acid addition salt (such as an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid or phosphoric acid or an organic acid such as methanesulfonic acid or toluene-p-sulfonic acid) )
or N-Cyryl I! The conductor or 4-position is of the formula -〇〇OR5 (formula R5 is a hydrogen atom or a carboxyl blocking functional group, such as an ester-forming aliphatic alcohol or an araliphatic alcohol or an ester-forming phenol, silanol or stannanol (this alcohol , a phenol, a silanol or a stannol having preferably 1 to 20 carbon atoms) and an associated group such as a halide (e.g. chloride or bromide) or a trifluoroacetate anion. A corresponding compound having an anion 80 of the formula [where R and Rb have the above definitions, R6 represents a carboxyl occluding group, for example the group described for R5 and R7 is an amino group or a protected is an amino group] or a corresponding acylating agent, or acylated with an acid or a corresponding acylating agent of the formula (B) [where R, Rb, R7, B and σ dotted lines have the above definitions; R8a can independently represent hydrogen or a carboxyl occluding group and X is a displaceable residue of a nucleophile, such as an acetoxy group or a dichloroacetoxy group, or a halogen atom, such as, for example, chlorine, bromine or iodine. ) or a salt thereof with a pyridine compound of the formula in which R4 has the above definition, followed by the following reactions in any suitable order, i) as desired, as required and/or desired in each case. Conversion of Δ2-isomer to Δ3-isomer of, 1i
) reduction of a compound in which B is 〉S→0 to produce a compound in which B is 〉S; 111) to a non-toxic salt of a carboxyl group or to a non-toxic metabolically labile ester function; and iV) removal of the carboxyl occluding group and/or the N-protecting group or a non-toxic salt thereof or a non-toxic antibiotic compound of the general formula (I) as defined above. A method for making metabolically unstable esters is provided.

In the method (A), the starting material of formula (I[) is preferably a compound in which B is >S and the dotted line represents a cef-3-em compound. One of the starting materials that has been found to be particularly suitable for use in process (A) due to the high purity it can produce is N-(7-aminocef-3-
Em-3-ylmethyl)pyridinium-4-carpoxylate dihydrochloride.

Examples of acylating agents which can be used for the preparation of compounds of formula (I) include, for example, acid halides, especially acid chlorides or acid bromides.

Such acylating agents can be prepared by reacting acid (I[) or a salt thereof with a halogenating agent such as phosphorus pentachloride, thionyl chloride or oxalyl chloride.

Acylation with an acid halide, optionally in the presence of an acid binder, is conveniently carried out at -50 to +50°C, preferably from -20°C to
It can be carried out in aqueous and non-aqueous reaction media at temperatures of +30°C. Examples of suitable reaction media include, for example, aqueous ketones (e.g. aqueous acetone), esters (e.g. ethyl acetate), halogenated hydrocarbons (e.g. methylene chloride), amides (e.g. dimethylacetamide), nitriles (e.g. acetonitrile) or two or more. Mixtures of more such solvents may be mentioned. Examples of suitable acid binders include, for example, tertiary amines (for example triethylamine or dimethylaniline), inorganic bases (for example calcium carbonate or sodium bicarbonate) and for example lower , 2-alkylene oxides (e.g. ethylene oxide or propylene oxide).

Acids of formula (I) are themselves used as acylating agents in the preparation of compounds of formula (I).

Acylation using an acid (Ill) is performed using a condensing agent such as N.

Carbodiimides such as N'-dicyclohexylcarbodiimide or N-ethyl-N'-γ-dimethylaminopropylcarpodiimide, carbonyl compounds such as carbonyldiimidazole or e.g. N-ethyl-5-phenylisoxazolium verchlorate. It is preferable to carry out the reaction in the presence of an isoxazolium salt such as

Acylation also includes other amides of acids of formula (III), such as activated esters, symmetrical anhydrides or mixed anhydrides (e.g. formed with pivalic acid or with haloformates such as lower alkyl haloformates). It can also be carried out using formable derivatives. Mixed anhydrides are phosphoric acid (e.g. phosphoric acid or phosphorous acid), sulfuric acid or aliphatic or aromatic sulfones! ! (e.g. toluene-p-sulfonic acid). The activated ester is conveniently produced in a reaction system using, for example, 1-hydroxybenzothiazole in the presence of the condensing agent mentioned above. Alternatively, the activated ester may be preformed.

Acylation reactions involving the free acids or their aforementioned amidogenic derivatives are preferably carried out in anhydrous reaction media such as methylene chloride, tetrahydrofuran, dimethylformamide or acetonitrile.

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

Acids of formula (III) and their corresponding acylating agents can optionally be produced and used in the form of their acid addition salts. In other words, for example, acid 0 ride is that! ! j! jJ
! and the acid bromide is conveniently used as its hydrobromide salt.

The pyridine compound of formula (V) can act as a nucleophilic reagent to substitute a wide variety of substituents X from the cephalosporin of formula (IV). To some extent, the ease of substitution is related to the pKa of the IIHX into which the substituent is derived. That is, atoms or groups X derived from strong acids generally tend to be more easily substituted than atoms or groups derived from weaker acids. Also, the ease of substitution is to a certain extent the formula (V
) is also related to the nature of the substituent R4 in the compound.

1llW! of X by the pyridine compound of formula (V)! is conveniently carried out by maintaining each reaction component in solution or suspension. The reaction is advantageously carried out using 1 to 10 mol of pyridine compound.

The nucleophilic substitution reaction is conveniently carried out on compounds of formula (IV) in which the substituent X is, for example, a halogen atom or an acyloxy group as described below.

Compounds of formula (IV) in which X is an acetoxy group are convenient starting materials for use in nucleophilic substitution reactions with pyridine compounds of formula (V).

Examples of other starting materials of this type include, for example, the formula (TV
) in which X is the residue of a substituted acetic acid, such as chloroacetic acid, dichloroacetic acid and trifluoroacetic acid.

The substitution reaction for compounds (mV) with X substituents of this type, especially when X is an acetoxy group, can be accelerated by the presence of iodide or thiocyanate ions in the reaction medium. This type of reaction is described in British Patent No. 1,132,621 (Patent No. 757847) and British Patent No. 1,171,603 (Patent No. 63458).
No. 4) Detailed information is provided in each specification.

Substituent X may also be derived from formic acid, haloformic acid (for example, formic acid) or carbamic acid.

When using a compound in which X represents an acetoxy group or a substituted acetoxy group in formula (IV), generally the formula <IV
) is preferably a hydrogen atom and B represents >S. In this case the reaction is preferably carried out at a pH of 5 to 8.
It is particularly advantageous to carry out the process in an aqueous medium at a pH of 5.5 to 7.

The method using compounds of formula (rV) in which X is the residue of a substituted acetic acid may be carried out as described in GB 1,241,657 (Patent No. 833,410).

When using a compound in which X is an acetoxy group in formula (IV), the reaction temperature is 30° to 110°C, preferably 50°C.
Conveniently, it is carried out at a temperature of ~80°C.

Compounds of formula (IV) in which X is a chlorine atom, bromine atom or iodine atom are also conveniently used as starting materials in the nucleophilic substitution reaction with the pyridine compound of formula (V). When using compounds of formula (rV) of this type, B can represent 〉S→0 and R8 a carboxyl-occluding functional group. The reaction is preferably carried out in one or more organic solvents, preferably polar ones, such as ethers (e.g. dioxane or tetrahydrofuran), esters W74 (e.g. ethyl acetate), amides (e.g. formamide and N,N- It is convenient to carry out the reaction in a non-aqueous medium consisting of dimethylformamide) and ketones (eg acetone). In some cases the pyridine compound itself can be a solvent. Other suitable organic solvents include British Patent No. 1,326,531
No. (Patent No. 874185) Mei 1B. The reaction medium must neither be extremely acidic nor extremely basic. In formula (IV) R8
and in the case of reactions carried out on compounds in which R8a is a carboxyl occluding group, the 3-pyridinium methyl product is produced as the corresponding halide salt, which is optionally subjected to one or more ion exchange reactions. A salt having a desired anion can be obtained.

When using compounds of formula (IV) in which X is a halogen atom as defined above, the reaction is conveniently carried out at a temperature of -10 DEG to +50 DEG C., preferably +10 DEG to +30 DEG C.

The reaction products can be removed, e.g., from unreacted cephalospores, by a number of methods, including, e.g., recrystallization, iontophoresis, column chromatography, and the use of ion exchangers (e.g., by chromatography on ion exchange resins) or macroreticular resins. The phosphorus can be separated from the reaction mixture containing the starting material and other materials.

The Δ2-cephalosporin ester derivative obtained according to the method of the invention can be converted into the corresponding Δ3-derivative, for example, by treating the Δ2-ester with a base such as pyridine or triethylamine.

The cef-2-em reaction product can also be reacted with, for example, a peracid such as peracetic acid or m-chloroperbenzoic acid to form the corresponding cef-3-em 1-oxide, the resulting sulfoxide being the desired sulfoxide. can be subsequently reduced to produce the corresponding cef-3-em sulfide as described below.

If a compound is obtained in which B is 〉S→0, this means, for example, that in the case of an acetoxysulfonium salt, the corresponding acyloxysulfonium salt or alkoxysulfonium salt prepared in the reaction system, for example by reaction with acetyl chloride, is obtained. It can be converted to the corresponding sulfide by reduction. The reduction is carried out, for example, with sodium dithionite or with iodide ions in a solution of potassium iodide in a water-miscible solvent such as acetic acid, acetone, tetrahydrofuran, dioxane, dimethylformamide or dimethylacetamide. The reaction can be carried out at temperatures between -20° and +50°C.

Metabolically unstable ester derivatives of compounds of formula (I) are prepared by preparing the compound of formula (I) or a salt or protected derivative thereof, conveniently in an inert organic solvent such as dimethylformamide or acetone, with an acyloxyalkyl halide ( It is prepared by reacting with a suitable esterifying agent such as iodite) and then removing the protecting group if necessary.

Base salts of compounds of formula (I) are formed by reacting acids of formula (I) with a suitable base.

Thus, for example, the sodium or potassium salts may be prepared using the 2-ethylhexanoate or bicarbonate salts, respectively. Acid addition salts may be prepared by reacting a compound of formula (I) or a metabolically labile ester derivative thereof with a suitable acid.

When the compound of formula (I>) is obtained as a mixture of isomers, the syn isomer can be obtained by conventional methods, such as crystallization or chromatography.

For use as starting materials for the preparation of the compounds of general formula (I) according to the invention, the syn isomer and the corresponding anti-isomer are in the syn isomer form or contain at least 90% of the syn isomer. Preference is given to using compounds of the general formula (I[[) and the corresponding acid halides and acid anhydrides in the form of mixtures of.

The acid of formula (■) (provided that R and Rb do not form a cyclopropylidene group together with the carbon atom to which they are bonded) is an acid of the formula [wherein R7 has the above definition]. and R9 represents a carboxyl occluding group. can be prepared by reaction with a compound such as a sulfonate, etherification, and subsequent removal of the carboxyl-occluding functional group R9.

Separation of isomers can be carried out either before or after such etherification. The etherification reaction is generally carried out in the presence of a base such as potassium carbonate or sodium hydride and an organic solvent such as dimethyl sulfoxide, a cyclic ether (such as tetrahydrofuran or dioxane) or N.

Preferably it is carried out in an N-disubstituted amide (eg dimethylformamide). Under these conditions, the configuration of the oximino groups is not substantially changed by the etherification reaction. The reaction should be carried out in the presence of a base when using acid addition salts of compounds of formula (Vl). The base should be used in an amount sufficient to rapidly neutralize the acid in question.

Acids of the general formula (I[
The carboxyl occluding action iR
9 and optionally separating the syn- and anti-isomers.

The last mentioned reaction is expressed in formula (I[) with Ra and Rb
are particularly applicable to the production of acids in which the cyclopropylidene groups together with their bonded carbon atoms form a cyclopropylidene group.

In this case, related compounds of formula (IX) may be used, for example, in Belgian Patent No. 866,422 for the preparation of tertiary-butyl 1-aminooxycyclopropane carboxylate.
It can be manufactured in a conventional manner by the synthetic method described in the specification.

Acids of formula (Ill) may be converted into the corresponding acid halides and anhydrides and acid addition salts by conventional means such as those described above.

When X in formula (IV) is a halogen (i.e. chlorine, bromine or iodine) atom, the cef-3-em starting compound can be prepared, for example, by a method analogous to 7β-
Protected amino-3-methylcef-3-em-4-carboxylic acid ester 1β-oxide is halogenated to form its 7β-
The vines are prepared in a conventional manner by removing the protecting group, acylating the resulting 7β-amino compound to produce the desired 7β-acylamido group, and reducing the 1β-oxide group later in the process. This is British Patent No. 1.326,5
It is described in the specification of No. 31 (Patent No. 874185). The corresponding cef-2-em compound can be prepared by reacting a 3-methyl cef-2-em compound with N-bromosuccinimide according to the method described in Dutch Patent Application No. 6,902,013. Methylcef
It can be produced by obtaining a 2-M-compound.

When X in formula (IV) is an acetoxy group, such a starting material can be prepared, for example, by acylating 7-aminocephalosporanic acid in a similar manner to method (A) above. Compounds in which X represents another acyloxy group in formula (TV) are disclosed in British Patent No. 1,474.
For example, a suitable 3-acetoxymethyl compound may be It can be produced by acylating the corresponding 3-hydroxymethyl compound which can be produced by hydrolysis.

The starting material of formula (II>) is also used in British Patent No. 1.028,5
63 (Japanese Patent No. 557165), it can be produced by a conventional method, for example, by subjecting the corresponding 3-acetoxymethyl compound to nucleophilic substitution with an appropriate nucleophilic reagent.

A further method for preparing the starting material of formula (I[) is, for example, PC
It consists of deprotecting the corresponding protected 7β-amino compound using N5 in a conventional manner.

It should be recognized that in some of the above transformations it is necessary to protect any reactive groups in the molecule of the compound to avoid undesirable side reactions. For example, during any of the above reaction steps the N82 group of the aminothiazolyl moiety may be tritylated,
Protection by acylation (eg chloroacetylation), protonation or other conventional methods may be necessary. The protecting group can then be removed by any convenient method that does not destroy the desired compound, for example in the case of trityl groups with optionally halogenated carboxylic acids such as acetic acid, formic acid, chloroacetic acid or trityl group. removed using fluoroacetic acid or using a mineral acid such as hydrochloric acid or a mixture of such acids, preferably in the presence of a polar solvent such as water or, in the case of chloroacetyl groups, by treatment with thiourea. It can be done.

The carboxy-occluding functional group used in the preparation of the compound of formula (I) or the necessary starting materials is preferably a group that can be easily cleaved off at an appropriate step of the reaction process, conveniently at the final step. However, in some cases non-toxic metabolically inactive groups such as acyloxymethyl or acyloxyethyl groups (for example acetoxymethyl or acetoxyethyl or pivaloyloxymethyl) are used to obtain suitable ester derivatives of compounds of formula (I). It is advantageous to use stable carboxyl-occluding functional groups and to retain these in the final product.

Suitable carboxyl occluding groups are well known in the art, and a list of representative blocked carboxyl groups is provided in British Patent No. 1,399.088 (Japanese Unexamined Patent Publication No. 48-4487). It is described in. Examples of preferred blocked carboxyl groups include, for example, aryl lower alkoxycarbonyl groups (e.g. p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl and diphenylmethoxycarbonyl), lower alkoxycarbonyl groups (e.g. tertiary butoxycarbonyl) and Examples include lower haloalkoxycarbonyl groups (for example, 2.2.2-trichloroethoxycarbonyl). The carboxyl occluding group may then be removed by any suitable method disclosed in the literature. Thus, for example, in addition to enzyme-catalyzed hydrolysis, acid- or base-catalyzed hydrolysis can be applied in many cases.

The antibiotic compounds of the invention, like other antibiotics, can be formulated for administration in any convenient manner and the invention therefore falls within its scope for use in human or veterinary medicine. Includes pharmaceutical compositions containing adapted antibiotic compounds according to the invention. Such compositions may be provided for use in conventional manner with the aid of any necessary pharmaceutical carriers or excipients.

Antibiotic compounds according to the invention can be formulated for injection and presented in unit dosage form in ampoules or multi-dose containers, optionally with an added preservative. The compositions can also take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles and contain formulation agents such as clouding agents, stabilizing and/or dispersing agents. I can do it.

Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, eg sterile pyrogen-free water, before use.

Optionally, such powder formulations may be formulated to improve the water solubility of the active ingredients and/or to ensure that the pH of the resulting aqueous formulation when the powder is reconstituted with water is physiologically acceptable. It may contain a suitable non-toxic base. Alternatively, the base may be present in the water for reconditioning the powder.

The base is, for example, 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 of the present invention may also be prepared as a herbal medicine containing a herbal base such as cocoa butter or other glycerides.

Veterinary pharmaceutical compositions may also be formulated as, for example, mammary gland preparations, either long-acting or rapid-release.

Depending on the method of administration, the composition may be 0.1% or more, e.g.
May contain 99% active substance. When the composition consists of dosage units, each unit preferably contains from 50 to 1500 jt9 of active ingredient. The dosage used for the treatment of adults depends on the route and frequency of administration, but may be up to 50 ml per day.
0 to 6000 is preferable. For example, in the treatment of adults, 1000-300 mg per day for intravenous or intramuscular administration
000η will normally be sufficient.

Higher daily doses may be required for the treatment of pseudomonas infections.

Antibiotic compounds according to the invention may be administered together with other therapeutic agents, such as antibiotics such as penicillin or other cephalosporins.

Next, the present invention will be explained by examples. All temperatures are in °C
It is. "Betrol" is petroleum ether (boiling point 40-6
0°).

Proton magnetic resonance (p, m, r,) spectrum is 100MH
Measured in z. Each integral corresponds to an assignment and is expressed by a coupling constant JLtHz, and each sine or S
- single line, d - doublet, dd - overlapping doublet, m - polyperpendicular and ABq - AB quartet not measured.

Production method 1 Ethyl (Z)-2-(2-aminothiazole-
4-yl)-2-(hydroxyimino)acetate A solution of ethyl acetoacetate (292 g) in wood vinegar 1 (296IIi) was stirred and ice-cooled at a rate such that the reaction temperature was maintained below 10°C. Water (400
A solution of sodium nitrite (1809) in ae) was added. Continue stirring and cooling for approximately 30 minutes, then add potassium chloride (1601J) in water (800d).
) solution was added. The resulting mixture was stirred for 1 hour. The lower oily phase was separated and the aqueous phase was extracted with diethyl ether. The extracts were combined with an oil, washed successively with water and saturated brine, dried and evaporated. The residual oil, which solidified on standing, was washed with betrol and then dried in vacuo over potassium hydroxide to give ethyl (Z)-2-
(Hydroxyimino)-3-oxobutyrate (309
17) was obtained.

Ethyl (Z)-2- in dichloromethane (400 ae)
(Hydroxyimino)-3-oxobutyrate (150
The stirred and ice-cooled solution of 9) was treated portionwise with sulfuryl chloride (140 g).

The resulting solution was kept at room temperature for 3 days and then evaporated. The residue was dissolved in diethyl ether, washed with water until the washings were approximately neutral, dried and evaporated. The residual oil (177 g) was dissolved in ethanol (500 -) and dimethylaniline (77J121!) and thiourea (42 g) were added with stirring.

After 2 hours the product was collected by filtration, washed with ethanol and dried to give the title compound [73 g, mp 188°
(decomposition)] was obtained.

Production method 2 Ethyl (Z)-2-hydroxyimino-2-(
2-Tritylaminothiazol-4-yl) acetate salt (12,91) in dimethylformamide (28d) containing triethylamine (8,4d).
Stir and cool the solution of 9) (-30'L) for 2 hours or once with trityl chloride (16,75'J).
) was added dropwise. The mixture was allowed to warm to 15" for 1 hour, stirred for a further 2 hours and then partitioned between water (500d) and ethyl acetate (500Id'). The organic phase was separated and washed with water (2x500d). Shitsuide I NHC
jl (500d). Collect the precipitate and add water (10
0d), washed successively with ethyl acetate (200ate) and ether (200d) and then dried in vacuo to give the title compound as a white solid (16.4g, mp 18.0d).
4-186' (decomposition)).

Preparation 3 Preparation of Ethyl (Z)-2-(2-tert-butoxycarbonylprob-2-oximino)-2-(2-tridel-aminothiazol-4-yl)acetate in dimethyl sulfoxide (200 dl) A solution of tertiary-butyl 2-bromo-2-methylpropionate (2511I!) in dimethyl sulfoxide (2511I!) was stirred under nitrogen.
24,5 g> and potassium carbonate (34,6 eJ) were added and the mixture was stirred for 6 hours at room temperature. The mixture was poured into water (2 Jl), stirred for 10 minutes and filtered.

The solid was washed with water and dissolved in ethyl acetate (600rd). The solution was washed successively with water, 2N hydrochloric acid, water and saturated brine, dried and evaporated.

The residue was recrystallized from petroleum ether (boiling point 60-80°) to give the title compound [34 g, mp 123.5-125°
] was obtained.

Production method 4 (Z)-2-(2-tert-butoxycarbonylprop-2-oximino)-2-(2-t-cutylaminothiazol-4-yl)acetic acid The product (2 g) of production method 3 was dissolved in methanol ( 20m) and to this was added 2N sodium hydroxide (3,3ad). The mixture was refluxed for 1.5 hours and then concentrated. The residue was taken up in a mixture of water (50m), 2N hydrochloric acid (7dl) and ethyl acetate (50d). The organic phase was separated and the aqueous phase was extracted with ethyl acetate.

The organic solutions were combined and washed sequentially with water and saturated brine;
Dry and evaporate. The residue was recrystallized from a mixture of carbon tetrachloride and betrol to give the title compound (19
, melting point 152-156° (decomposition)].

Production method 5 Ethyl (Z)-2-(2-tritylaminothiazol-4-yl)-2-(1-M tertiary butoxyluponylcyclopto-1-oximino)acetate Product of production method 2 (55.8 g) Potassium carbonate (
The finely ground material, 31.29) was stirred in dimethyl sulfoxide (400 d') under nitrogen. After 30 minutes, tertiary butyl 1-bromocyclobutanecarboxylate (29,29) was added. After 8 hours, more potassium carbonate (31,29) was added. Over the next 3 days more potassium carbonate (6X 16g portions) was added and after 3 days more tert-butyl 1-bromocyclobutanecarboxylate (3.45g) was added.

After a total of 4 days the mixture was poured into ice-water (approx. 3j) and the solids were collected by filtration and washed thoroughly with water and Betrol. The solid was dissolved in ethyl acetate and the solution was washed with brine (twice), dried over magnesium sulfate and evaporated to give a foam. This foam was dissolved in ethyl acetate-vetrol (1:2) and filtered through silica gel (500 g). Evaporation gave the title compound (60i) as a foam. vmaX(C
HBr 3) 3400 (NH) and 17303-' (
ester).

Production method 6 <Z)-2-(1-tert-butoxycarbonylcyclobut-1-oximino)-2-(2-trityl-aminothiazol-4-yl)acetic acid The product of production method 5 (3.2 g) and Potassium carbonate (1,
A mixture of 659) was refluxed in methanol (180d) and water (20id) for 9 hours and the mixture was cooled to room temperature. The mixture was concentrated and the residue was partitioned between ethyl acetate and water, 2N HCj! (12,2m) was added. The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic extracts were combined, washed with saturated brine, dried and evaporated to give the title compound (2.3 g
) was obtained. λmax 1% (ethanol) 265M (El, 243).

Preparation 7 (Z)-2-(1-tertiary-butoxycarbonyl-pro-1-oxyimino)-2-(2-tritylaminothiazol-4-yl)acetic acid hydrazine hydrate in methanol (0,4d) (
A solution of 0,20 g) of 1-tert-butoxycarbonylcyclobut-1-oxyphthalimide (0,619, Belgian Patent No. 866) in dichloromethane (7d)
．． No. 422). The mixture was stirred for 1 hour at room temperature and then treated with 5N aqueous ammonia (7-).

Separate the organic phase and dissolve the aqueous phase in dichloromethane.
- Extracted. The organic solutions were combined, washed with water, dried and evaporated. The oily residue (0.30 g) was dissolved in a mixture of ether <5 m) and ethyl acetate (5 ae). 2-tritylaminothiazol-4-ylglyoxylic acid (0,73 g, Belgian Patent No. 864,
828) was added. The mixture was stirred overnight at air temperature and then filtered. The solid was washed with a small amount of ether and dried in vacuo to give the title compound (o, sg, mp 156.8-157.2').
I got it. v max (CHBra) 2300-350
0 (Q-@, N-H), 1750 (tertiary butyl ester), 1690. -' (acid).

Production method 8 Ethyl (Z)-2-(1-tertiary butoxycarbonylcyclobent-1-yloxyimino)-2-(
2-Tritylaminothiazol-4-yl)acetate The product of Preparation 2 (IOg) was purified by tertiary-butyl 2-promosilane in dimethyl sulfoxide (40d) containing potassium carbonate (10g) for 21 hours at 21° under nitrogen. Stirred with clopentane carboxylate (7g). The mixture was poured into ice-water (500 mft) and the gray solid was collected by filtration, washed with water and air dried.

This solid was recrystallized from methanol (soo,e) to give the title compound (11.7 g, mp 179-180°). vmax (CHBr a ) 3410 (NH)
, 1735 (ester) , 1275 (ester)
and 755 value −1 (phenyl).

Production method 9 (Z)-2-(1-tertiary butoxycarbonylcyclobent-1-yloxyimino)-2-(2-
The product of trityl-aminethiazol-4-yl)acetic acid Preparation 8 (625a+y') was refluxed for 7 hours with 2N sodium hydroxide solution in methanol (0.5d) and water (11J1). The mixture was allowed to cool overnight. After dilution with water, orthophosphoric acid is added to bring the solution to pH.
Adjusted to 2. The precipitate was extracted with ether and the combined extracts were washed with brine.

After drying with magnesium sulfate and evaporating the solvent, the gum (49
3■) was obtained. Recrystallization from diisopropyl ether gave the title compound (356η, mp 171-173°). vmaX (CHBr 3) 2500-3500
(OH and NH), 1755 (ester), 169
2 (acid) and 755 and 770IJ-1 (phenyl).

Production method 10 (6R,7R)-7-amino-3-(1-
(Viridinium methyl)cef-3-em-4-carboxylic acid dihydrochloride (a) (6) in dichloromethane (30m)
R97R>-7-(2-chenylacetamide)-3-
Stirring of (1-pyridinium-methyl)cef-3-em-4-carboxylate (4.15 g)! N the cloudy liquid,
Treated with N-dimethylaniline (5.09m) and chlorotrimethylsilane (2.52m).

The mixture was stirred for 1 hour at 30-35° and then
6, treated with phosphorus pentachloride (4,16'9), stirred for an additional hour at -25° to -306, and then mixed with dichloromethane (20d) and butane-1,
Poured into a stirred cooled (-20') solution of 3-diol (8.1m). The solution was left to reach a temperature of Oo for 30 minutes and the precipitated solid (A) was filtered, washed with dichloromethane and dried in vacuo. It was redissolved in methanol (17.5 mg), stirred and diluted with dichloromethane (87.5 m) and the precipitated solid was collected, washed with dichloromethane and dried in vacuo to give the title compound as white. Obtained as a solid (3.2 g). λmax (pH6 buffer) 258 darkness (E'23)
18),'l: (D20) value Gt O,95,1,3
2 Oyohi 1.84 (pyridinium proton), 4.10
~4.46 (ABQ, J16H2, 3-CH2-)
, 4.56 (d, J5H2,7-H), 4.70 (
d, J5Hz, 6-H), 6.14-6.50 (
ABQ, J17Hz, C2-H).

(b) 1 of the solid (A) produced in step (a) above.
Dissolved in N hydrochloric acid (25rd'). Isopropanol (95d) was added to precipitate the crystalline title compound as a hydrochloride (4.95g). τ(020) value is 1
．． 02. 1.36 and 1.87 (pyridinium protons), 4.2+ 4.55 (ABQ, J=
14H2.

3-CH2-), 4.62 (d, J=5H7.

Cy-H), 47.4(d, J=5Hz, Go
'-H), 6.19 + 6.38 (A'BQ,
J-18Hz, 02-H). The water content by Karl Fischer method is 9.4%.

Example 1 a> Tertiary butyl (6R,7R)-3-acetoxymethyl-7-[(Z) -2-(2-tert-butoxycarbonylprobu-2-oximino)-2-(2-trityl aminothiazol-4-yl)acetamido)ceph-
3-M-4-carboxylate The product of Preparation 4 (5721117> and tertiary butyl (6R17) in dimethylformamide (10m)
A stirred solution of R)-3-acetoxymethyl-7-aminocef-3-em-4-carboxylate (328Rg) was cooled to 0° and treated with 1-hydroxybenzotriazole (150II1g) followed by dicyclohexylcarbodiimide ( 225III!il) was added. The mixture was warmed to room temperature, stirred for 5 hours and then left overnight. The mixture was filtered and the white solid was washed with a small amount of ether. The furnace liquor and washings were diluted with water (50 m2) and extracted with ethyl acetate. Combine the organic extracts and add water,
It was washed successively with 2N hydrochloric acid, water, sodium bicarbonate solution and saturated brine, dried and evaporated. The residue was eluted through a silica column with ether. The product-containing eluate was collected and concentrated to give the title compound (533■)
I got it. A portion was recrystallized from diisopropyl ether. Melting point 103-113° (decomposition), (α)'F 8
．． 5' (c, 1.O, DMSO).

b) (6R,7R)-3-acetoxymethyl-7
-[(Z)-2-(2-aminothiazol-4-yl)
-2-(2-carboxyprop-2-oximino)acetamido)cef-3-em-4-carboxylic acid 06 in anisole (18d) above a)
To a solution of 2.4 g of the product (1! (1
8m> was added. 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 pH of the aqueous extract was adjusted to 6 and the solution was washed with ethyl acetate. The aqueous phase was acidified to pH 1.5 under ethyl acetate, saturated with sodium chloride and extracted with ethyl acetate. Combine the organic extracts, wash this with saturated saline,
Dry and evaporate. The residue was dissolved in a 50% warm aqueous solution (20ml) and allowed to stand for 2 hours. The mixture was diluted with water (50ml) and filtered. The filtrate was concentrated. The residue was dissolved in water (50ml). The title compound (920) was obtained by filtering again and freeze-drying.

λWaX (pH 6 buffer) 236 pieces (El: 250
), λinf 255/11 (235 to E), 29
6n! A (103 to E), c) (6R, 7R)
-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide)-3-(1-pyridinium methane)
- Cef-3-em-4-carboxylate monosodium salt pyridine <2d) and sodium iodide (7,12g) in water (2,2m>) and the product of b) above (1,8g) at 80° The solution was stirred for 1 hour at 80° C., cooled and diluted with water to 100 II 11. The pH of the solution was adjusted to 6.0 with 2N sodium hydroxide solution.
0 and the solution was concentrated to remove pyridine. The aqueous residue was diluted to 100 d with water, to which methyl isobutyl ketone (2 drops) was added and the solution was acidified to pH 1 with 2N hydrochloric acid.

The mixture was washed with a small amount of water.

The furnace liquor and washing liquor were collected and washed with ethyl acetate and the pH was adjusted to 6.0 with 2N sodium hydroxide solution.

The solution was concentrated to 50 μm and applied to a 500 g column of Amberlite XAD-2 resin using first water and then 20% aqueous ethanol as eluents. The product containing fractions were concentrated and lyophilized to give the title compound (0,569). λwax (pH 6 buffer 7
- ) 253.5 layers (El: 307), λinf
282 rat (159 to E), 2B0ru11 (
E'+', 295), [α)D+24.5' (C
1,0°DMSO).

Example 2 (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprob-2
-oximino)acetamido)-3-(4-carbamoyl-1-pyridiniummethyl)cef-3-em-4-
Carboxylate monosodium salt Water containing sufficient sodium bicarbonate (0,7 td)
Isonicotinamide (o, se p ) was added to a stirred solution of the product of Example 1b) (0.59 g) in a final 1)H of 6.5. Sodium iodide (2,1S
F) was added and the mixture was stirred for 1 hour at 80°C, to which sodium bicarbonate was added from time to time to maintain the pH between 5.5 and 6.5. The product was isolated essentially as described in Example 1G) to give the title compound (0.09 g). λ1aX (pH6 buffer) 1% 257.5rat (E 1.276), λinf 2
91.5 nm (El: 125), τ (D20) value is 0
．． 92.1.70 (4H1 pyridinium proton), 3
．． 10 (IH.

aminothiazole-5-H), 4.34. 4.64 (
2H, ABo, 3-CI-12-), 8.54 (6
H.

-CMe2-).

Example 3 a) Tertiary butyl (6R,7R)-3-acetoxymethyl-7-[(Z)-2-(1-tert-butoxy-carbonylcyclobut-1-oximino)-2-(2 -tritylaminothiazol-4-yl)acetamido)cef-3-em-4-carboxylate product of Preparation 6 (24,
2 g> and tert-butyl (6R17R)-3-acetoxymethyl-7-aminocef-3-em-4-carboxylate (13,6 g) was cooled to Oo and 1
-Hydroxybenzothiazole monohydrate (4,5 g) was then treated with dicyclohexylcarbodiimide (6,49) and the product was isolated essentially as described in Example 1a) to give the title compound ( 12,89), melting point 11
3.5-116.5° (decomposition) was obtained. [α]v+1
5.0' (and 1.0.DMS○).

b) (6R,7R)-3-acetoxymethyl-7
-[(Z)-2-(2-aminothiazol-4-yl)
-2=(1-carboxycyclobut-1-oxyimino)acetamideracef-3-em-4-carboxylic acid In 06, the mixture of the product (12,59) of above a) and anisole (5d) was added with trifluoroacetic acid. (100
ne) was added. The mixture was worked up essentially as described in Example 1b> to give the title compound (4g). 2iax
(pH-15 buffer -> 2464 (El, 26
4>, λinf 295 M (E: lj upper 11B),
1% [α]pi+27.3' (dan 1.0. DMSO).

c) (6R,7R)-7-((2)-2-(aminothiazol-4-yl)-2-(1-carboxycyclobuto-1-oximino)acetamide)-3-(1-
Pyridinium methane-ceph-3-em-4-carboxylate monosodium salt pyridine (4,1d) and the product of b) above (3,7
59) was added to a stirred solution of sodium iodide (14.6 g) in water (4.5 m) at 80° C. and the product was isolated essentially as described in Example 1c) to give the title compound (1 , 3g) was obtained. λ1Ilax(1)H6 buffer) 252.5M(E'j !310),
λinf 291 nIR (E 1% 139), (α
) pH 43.5"G (1.0.DMSO).

Example 4 (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclobuto-1-oximino)acetamide)-3-(carbamoyl- 1-pyridiniummethyl)cef-3-em-4-
Carboxylate monosodium salt To a stirred solution of the product (1,089) of Example 3b) in water (1,3d) containing sufficient sodium bicarbonate isonicotinamide (1,22 g) is added to give the final p.
H was set to 6.5. Sodium iodide (4 g) was added and the mixture was stirred for 1 hour at 80°C until the p.
Sodium bicarbonate was added occasionally to maintain H.

The product was isolated essentially as described in Example IC> to give the title compound (0,169).

Obtained. (α) D-ia (Dan 1.08, H20)
, 2iax (pH 6 buffer) 256M (E
', 2298), λinf 294 run (E
1%135).

1α Example 5 a) Tertiary butyl (6R,7R)-3-7cetoxymethyl-7- [(Z)-2-<2-tritylaminothiazol-4-yl)-2-(1-tertiary butoxy Preparation of carbonylcycloprop-1-oxyimino)acetamidoracef-3-em-4-carboxylate 1-hydroxybenzotriazole monohydrate (0,12g) and dicyclohexylcarbodiimide (0,169) in tetrahydrofuran (6d) 7 product (0,3
4g) and tert-butyl (6R,7R)-3-acetoxymethyl-7-aminocef-3-em-4-carboxylate (0,257). The mixture was stirred overnight at room temperature and then filtered. The furnace liquid was evaporated.

The residue was dissolved in a small amount of ethyl acetate-petroleum ether (boiling point 60~
80°) (1:1) and eluted through a column of neutral alumina (10 g) using the same solvent.

The eluate was concentrated to give a form (0.44g) which was recrystallized from diisopropyl ether (15d) to give the title compound (0.29g). Melt b) (6R,
7R)-3-acetoxymethyl-7-[(Z)-2-(
2-aminothiazol-4-yl)-2-(1-carboxycycloprop-1-oximino)acetamide)
In Cef-3-M-4-carboxylic hydrochloride io'? l Hydrochloric acid (0.6 mg) was mixed with formic acid (7.5 mg).
m) to a stirred solution of the product from a) above (1.92 g) in m). The mixture was stirred for 1.25 hours at room temperature and then filtered. Diisopropyl ether (30%
0 m > and the mixture was stirred for 1.5 hours. The solid was quenched, washed with diisopropyl ether and diethyl ether and dried in vacuo to give the title compound (1.16g).

(α) 20 (c 1.0. DMSO) +35°,
λmax (pH6 buffer) 239 questions (12300)
.

c) (6R,7R)-7-[(Z)-2-(2-
aminothiazol-4-yl)-2-(1-carpoxycyclopObo-1-oximino)acetamide)-3
-(1-pyridiniummethyl)cef-3-em-4-carboxylate sodium salt A mixture of the product from b) above (0,56 g), sodium bicarbonate (o,17 g) and water (0,5 alt) 50 m.Furthermore, sodium hydrogen carbonate (
0,091J) was added, then pyridine (0,2m
> was added. Humidify the solution at 80° and add sodium iodide (
2g) was added. The solution was stirred for 40 minutes at 806, cooled and diluted with acetone (50m). The mixture was filtered and the solid was washed with acetone and ether to obtain a solid. This solid was dissolved in water (20d) and acidified in portions with 2N hydrochloric acid until a precipitate formed which did not dissolve again on standing. The mixture was stirred with neutral alumina (5g) and filtered through a pad of neutral alumina (10g). The pad was completely eluted with water. The aqueous eluate was concentrated and the residue was triturated with acetone. The solid was filtered and dried to give a solid (0.35g). This solid (0,30!?>) was dissolved in a small amount of water and eluted through a column of 50 g Amberlite The product-containing fractions were concentrated and the residue was triturated with acetone to give the title compound (o, oeg).

[α]vO″±1.5” (c O, 1, water), λm
ax (pH 6 buffer y -) 254ruR (E 1
:4340), λ inf 296 nxr (
E cost 125n.

Example 6 (6R,7R)-7-[(Z)-2-aminothiazol-4-yl)-2-(1-carboxycyclobent-1
-yl-oximino)-acetamide)-3-(1-
Pyridinium methyl) cef-3-em-4-carboxylate dihydrochloride phosphorus pentachloride (0,469>) was dissolved in methylene chloride (20 m) at ambient temperature and the solution was cooled to 10° to produce Preparation 9. (1,095g) 1
Added in the episode. The mixture was warmed to -5° and stirred for 30 minutes.

The solution was cooled to -10° and triethylamine (0,61-) was added followed by water (6,7-) with vigorous stirring so that the temperature was not above 0° but the water did not freeze.

The two-phase mixture was stirred for 3 minutes and transferred out of the tap oven.

The lower phase was heated with N,N-dimethylacetamide (1.4 m) containing triethylamine (1.4 m), previously cooled to -20°.
0d) and acetonitrile (@@10a) in 10 #l to a vigorously stirred suspension of the product (0.76 g), the addition being carried out at a temperature of -10"
This was done in such a way that it did not become too much. The mixture -5~-
Stirred at 10° for 45 minutes and left to warm to 21″ for 1 hour.

Methanol (0.3 m) was added and the methylene chloride was evaporated under reduced pressure at a bath temperature of 30°.

The residue was carefully partitioned between ethyl acetate (30m>) and water (30m>) and a small amount of sodium chloride was added. The first layer was washed with another portion of water (2 x 30m>). The combined sodium chloride was extracted with ethyl acetate (20d) and the organic layers were combined and dried over magnesium sulphate. Evaporation gave a foam (1.79g) which was diluted with diisopropyl ether. Trituration gave a solid (1.35 g).

Gently remove most of this solid (1.2g). (5 m) and concentrated hydrochloric acid (0.38 m) was added with vigorous stirring. After standing for 1 hour at 21°, the suspension was evaporated and the residue was leached with a little acid. The filtrate liquors were combined, concentrated by evaporation and the residue was triturated with acetone to give the title compound (374IRg).

(α) D+8.6' (c 1.02, H20
) λmax (pH6 buffer) 255IuR (E1%
289), λinfcm 1% 295 (E:, :273), λinf 280 (E
1.158).

Example 7 (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2
-oximino)acetamide)-3-(1-pyridiniummethyl)-cef-3-em-4-carboxylate sodium salt (6R,7R)-7-[(Z)-2-(2
-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide)-3-(
1-Pyridinium methyl)-cef-3-em-4-carboxylate (2,59) was dissolved in water and the solution was treated with sodium 2-ethylhexanoate (1,52 g) in methanol (8-). did.

The mixture was added to stirred acetone for 15 minutes and the resulting suspension was filtered, washed and dried in a dry rack c 1.0.
H20) , λmax (DH6 phosphate) , 2
55 (E near: 327, ε18630), λinf
2401% 1% (E 305, ε17,370) and 280 (E
172.1 cIRl cm 89, 800), νmaX (nusiE-le) 1780
Ca+-1 (β-lactam), sodium actual value: 4
．． Calculated for 5%, C22H2107N6S2N: 4.04%.

Example 8 a) Diphenylmethyl (I3.6R,7R)-7-[
(Z)-2-(2-tertiary butoxycarbonyl prop-
2-oximino) = 2- (2-tritylaminothiazol-4-yl)-acetamide 3-bromomethylcef-3-em-1-oxide-4-carboxylate Phosphorus pentachloride (0.75 g) was dissolved in methylene chloride. (20
The mixture was suspended while being stirred during the aeration. Stir the mixture to 10°
and the product of Preparation 4 (2.0 g) was added.

Stirring was continued for 10 minutes at -5 to -10°.

-io', triethylamine (0,88d) in methylene dichloride (5#11) was added, and then after 5 minutes, methylene dichloride (30ml) containing triethylamine (0,42d) was washed with methylene dichloride (5d).
') in diphenylmethyl (1S.

6R,7R17-amino-3-bromomethylcef-3-
A suspension of em-1-oxide-4-carboxylate hydrobromide (1.67 g) was added.

The mixture was stirred at -5 to -10° for 20 minutes and then poured into half-saturated aqueous sodium bicarbonate solution (50d). Separate the organic layer and add dilute hydrochloric acid solution (IN, 3X 30d
) and brine (2X 30d') and evaporated in vacuo to give a foam. The foam was taken up in ethyl acetate (approx. 10d) and treated with diisopropyl ether (100d). The precipitated solid was collected by filtration, washed with diisopropyl ether and dried overnight in vacuo at 40° to give the title compound (2.1 g). The value of τ(CDCl2) is 3°11(s, -ChlP
h2), 3.37(s, thia'1-)ti-5-yl'
70 tons) 3.88 (dd, J9Hz and 5H
z, 7-H), 5.22 + 6.02 (ABq-3
CH2), 5.49 (d, 5H2,6-H) 8.46 (
s, CMe 2 ).

b) (6R,7R)-7-[(Z)-2-(2-
aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide)-3-(1
-Pyridinium methyl)-cef-3-em-4-carboxylate The product of a) above (1 g) was dissolved in acetone (22 m) and stirred at room temperature. Pyridine (0.08m) was added and the mixture was stirred for 3 hours at room temperature. More pyridine (0.72d) was added and the mixture was left at room temperature overnight. The mixture was stirred with diethyl ether (75me
) and the precipitated solid was collected by filtration, washed with ether and dried in vacuo 401. This solid (0.8 g) was redissolved in acetone (22d) at -106. Potassium iodide (0.7 g) then acetyl chloride (0.17 g) was added. The mixture was heated to 10°C.
After stirring for 2θ minutes at
> and acetyl chloride (0.17m) were added.

After stirring for a further 20 min at -10°, the mixture was dissolved in water (60
d) and added to a solution of sodium metabisulphite (0.6 g) in saturated saline (3 hd!). The product was extracted with methylene dichloride (2X 5hte') and the extracts were washed with brine, dried over magnesium sulfate and evaporated under reduced pressure to give a foam.

This was dissolved in Gil (6,5%) and left at room temperature for 15 minutes. Concentrated hydrochloric acid (0.257) was added and the mixture was left for an additional 1.25 hours. The solid precipitate was filtered and washed with a small amount of phosphoric acid. The furnace liquor and washings were combined and poured into ethyl acetate (!M) and diethyl ether (5M) with water (10m) and acetonitrile (5Id).Additional water was added until two distinct layers were obtained. The lower layer was poured out, and then Amberlite LA2 (
7m) and diethyl ether (14m) containing acetic acid (0,7+d!). Separate the aqueous layer again and zeolite 225SRC15 (H+ form 15Id)
was applied to the column. The column was washed with water until neutral. The product was eluted with a 10% solution of pyridine in water.

The eluate was evaporated in vacuo to a small volume and treated with acetone. The mixture was cooled to 0-40° overnight and filtered. The solid was washed with acetone and dried in vacuo at 40° to give the title compound (0.25g). The nmr spectrum was similar to that of the compound prepared in Example 7. λl1laX (pH 6 phosphate > 255
．． 5/l#I (E'+! 374), 1% 1% λ1nf (E 1clR340) and 290 nm (
E −+ , 160).

Example 9 a) (6R,7R)-7-[(Z)-2-(2-
triphenylmethylaminothiazol-4-yl)-2
-(2-31-butoxycarbonylprop-2-oxyimino)acetamide)-3-(1-pyridinium-methyl)-cef-3-em-4-carboxylate Phosphorus pentachloride (
To a stirred solution (cooled to -10°) of 1,38 g) was added the product of Preparation 4 (3,44 g).The resulting solution was stirred for 30 minutes at -5° and then cooled to -10°.

Triethylamine (1.33 g) and Tweet water (20 d) were added. The mixture was stirred at Oo for 3 min and the lower phase was dissolved in a mixture of N,N-dimethylacetamide (30 m)/acetonitrile (30 m) containing triethylamine (3,03 g). 19g
) was added over 10 minutes to a stirred suspension of (cooled to -10°). The mixture was stirred for 45 minutes at -5'' and then for 11 hours without cooling. Methanol (Id) was added. Methylene chloride was removed by evaporation under reduced pressure. The remaining solution was stirred. The title compound (4.89 g) was precipitated in addition to water (300 d) while τCCDC.
ρ3) value is 2.78 (s.

-(Ca1-1s)3), 3.37 (S, -thiazole proton), 0.35, 1.80, 2
．． 12 (pyridinium flotone), 4.18 (m, -
7-)-1>, 4.95 (6-H), 8.66 (S
, -tertiary butyl), 8.50 (s.

-C(CHs)2).

b) (6R,7R)-7-[(Z)-2-(2-
aminothiazol-4-yl)-2-(2-carboxyprobu-2-oximino)acetamide)-3-(1
-Pyridinium methyl)-cef-3-em-4-carboxylic acid dihydrochloride The product from (a) above (3.38 g) was dissolved in 1 liter (20d) of 98% strength with stirring. Concentrated hydrochloric acid (1,
2 ml was added and the mixture was stirred for 1 hour. The precipitated solids were removed by vacuum filtration. The solvent was removed from the filtrate by evaporation under reduced pressure leaving an oil which was triturated with acetone (30m) to give the title compound (2.20g). The r (020/Na HCO3) value is 3.08 (s
, -thiazole proton), 1.06.

1.44. 1.93 (pyridinium proton), 4,
16(d, H5Hz, 7-1-1>, 4.74(d
, J 5 H2.

6-H), 8.55(s, -〇 (CH3) 2)
Contains. Acetone by n, m, r is 1 mole. Water content 5% (Karl Fischer method). Actual value of chlorine 10.1% [C22nd 24N, 07S2Cρ2+
Theoretical value of acetone (1 mol) soil water (5%) (:fJio
, o%] Example 10 a> (6,R,7R)-7-[(Z)-2-(2
-triphenylmethylaminothiazol-4-yl)-
2-(2-tert-butoxycarbonylprop-2-oximino)acetamide)-3-(1-pyridinium-
Example 9
Reaction as in (a) gave the title compound (4.03 g), whose spectroscopic properties were similar to those of the product of Example 9(a).

b) (6R,7R)-7-[(Z)-2-(2.
)-1'-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide)-3-(
1-Pyridinium methyl) cef-3-em-4-carboxylic acid dihydrochloride The product from (a) above (3,8 g>) was added to Example 9 (b).
The title compound (2.17 g) was obtained by treating as follows.
Its spectroscopic properties were similar to those of the product of Example 9(b).

Example 11 a> (6R,7R)-7-[(Z)-2-(2-
triphenylmethylaminothiazol-4-yl)-2
-(1-tert-butoxycarbonylcyclobut-1-oximino)acetamide)-3-(1-pyridiniummethyl)ceph-3-em-4-carboxylate phosphorus pentachloride (1,389) in 60-dichloromethane dissolved in it. The solution was cooled to -10° and the product of Preparation 6 (3,489) was added in one portion. solution for 30 minutes -
Stir at 5°. Triethylamine (1.8m) was added followed by water (20m). The mixture was stirred for 3 minutes at O<0>C. The lower phase was then treated with dimethylacetamide (30 m
) and acetonitrile (30d)
The product of 0a) (2°189) was added to the pre-chilled mixture.

The reaction mixture was stirred at -5°C to 10°C for 45 minutes.

Cooling was then removed and the reaction was stirred for an additional hour, during which time ambient temperature was achieved. The solvent was removed under reduced pressure and the residue was partitioned between ethyl acetate and water. The organic phase was washed with brine and the aqueous extracts were combined, which was extracted with ethyl acetate. Combine the ethyl acetate extracts,
This was dried in the presence of charcoal and the solvent was removed under reduced pressure. The residue was triturated with isopropyl ether to give the title compound (3.80 g). νmax (nujol)
1780cIR-' (β-lactam), r (CDC
jl s ) values are 2.74 (s, triphenylmethyl), 8.66 (s.

tertiary butyl).

b) (6R,7R)-7-[(Z)-2-(2-
aminothiazol-4-yl)-2-(1-carboxycyclo-but-1-oximino)acetamide)-3
-(1-Pyridiniummethyl)cef-3-em-4-carboxylic acid dihydrochloride The product from a) above (2,579) was dissolved for 1 hour in 98% formic acid (15m) and! Stirred in a mixture of hydrochloric acid (0,hte) at ambient temperature. The mixture was then filtered and the solvent removed under reduced pressure. The resulting residue was triturated with acetone to produce the title compound (1.79 g).

v maxc El -/L/) 1,785cm
-1 (β-lactam).

r (C20+Na HCO3) value is 1.05, 1
．． 42.

1.9Rm, pyridinium proton), 3.01 (s.

aminothiazole proton), 4.13 (d, J
5H2, Cy proton), 4.68(d, J5
Hz.

C-6 proton), 7.4-8.4 (broad m, cyclobutyl butoton).

dimethylacetamide (173
mol) and acetone (172 mol).

Water content 1.4% (Karl Fischer method).

Actual value of chlorine: 9.2% (C23H24N607S2Cρ
2+1/3 mol dimethylacetamide + 172 mol acetone + theoretical CI 9.5% as 7.4% water) Example 12 (a) (6R,7R)-3-acetoxymethyl-7
-[(Z)-2-<2-aminothiazol-4-yl'
)-2-(2-carboxyprop-2-oximino)
- Acetamidotucef-3-em-4-carboxylic hydrochloride Hydrochloride The product of Example 1(a) (200 g) was dissolved in pre-cooled acid (800 rd') to +10" and the stirred mixture thereof Added salt 112 (Sod) over 5 minutes to al. 20° to 22° for 1/4 hour
Stirring was continued at 20" and then cooled to +10" and filtered. The hearth was washed with 1 (30 ae). The furnace liquor and washing liquor were combined and this was evaporated at 20" and Ill! A yellow foam was obtained, which was dissolved in ethyl acetate (800 m >
Grinded with. The solid precipitated was collected by filtration, washed with ethyl acetate (200m2) and dried in vacuo at room temperature overnight to give the title compound (124.6g).

λmaX<eta/-)Lt)234.5nta, E
l:311°(b) (6R,7R)-7-[(Z)
-2-(2-aminothiazol-4-yl)-2-(2
-carboxyprop-2-oximino)acetamide)-3-(pyridinium-1-ylmethyl)cef-3-
Em-4-carboxylate hydrate Product from a) above (40g>) in water (40#11!
) and pyridine (25,6 kg) were added to a stirred mixture followed by sodium iodide (160 g) and the mixture heated to 60° for 3172 hours. The hot solution was poured into stirred acetone (2 O) and diethyl ether. (1,
2jl). The suspension was cooled to 2° and the crude product was collected by filtration (50.659). Dissolve this in water (480d) and then
, Amberlite L in ether (560d)
Stirred with A2 (280 m ). The mixture was separated and the organic layer was washed twice with water (24<17 m each). The aqueous layer was washed with ether (280 m ) and applied to a column of zeolite 225SRC15' (200 dH) before elution. The material was washed with distilled water until neutral. The column was eluted with 10% pyridine in water and the eluate was passed through a column of neutral alumina (40 g). The eluate was evaporated under reduced pressure to give a syrup. This syrup was added dropwise to stirred acetone (5007).

Filtration and equilibration in air yielded the title compound (13,0
9g') was obtained. 8207.0% (Karl Fischer), λmax 255nm (E', ', 364),
λ1nf1243 and 285. (i::i2338 and 171), [α]D13° (pH 6 phosphate buffer).

Example 13 (a) (6R,7R)-7-[(Z)-2-(2-
tritylaminothiazol-4-yl〉-2-(2-tertiary-butoxycarbonylprob-2-oximino)acetamide〕-3-(1-pyridiniummethyl)cef-
3-M-4-Carboxylate N,N-Dimethylformamide Solvate At 23° the micronized product of Example 9(a) was added to stirred N,N-dimethylformamide (15d). The solid dissolved and crystallization occurred after a while.

The stirred mixture was diluted by portionwise addition of diisopropyl ether (20d). The solid was collected by filtration to give the title compound (3.06 g) as colorless needles. N,N-dimethylformamide by n,m,r - 2172 moles.

"l: (DMSO-do): 2.4-3.0
(m, t-IJ methyl, 3.32 (S, aminothiazole ring proton), 0.47, 1.3B,
1.82 (pyridinium proton), C-6 proton)
, 8.64 (S, tertiary butyl proton), 8.6
2(s, (CH3)2-C/), [α)D--27
, 5° (c=i, i in methanol).

b) (6R,7R)-7-[(Z)-2-(2-
aminothiazol-4-yl)-2-(carboxyprop-2-oximino)-acetamide)-3-(1-
Pyridinium methyl) cef-3-em-4-carboxylic acid dihydrochloride The product (2,117) from a) above was dissolved in formic acid (10 #1
The mixture was melted at 22° in 1 hour. Concentrated hydrochloric acid (0.8d
) was added and after 75 minutes the precipitated solids were quenched. The furnace liquor was evaporated and industrial denatured alcohol (10d) was added. The solution was evaporated again. The residue was dissolved in methanol and the solution was added to diisopropyl ether to give the title compound (1.35 g).

(α]pi-14,7° (c-0,95, in pH 6 powder).

τ(DMSO-do): 0.28(d, J9.-
C-Nli), 0.77 (d, J6), 1.2
5(t, J6'), 1.70(t, J6')
, pyridinium ring proton>, 3.0 (s, aminothiazole proton), 3.99 (dd, J 9.5.
7-H), 4.67((j, J5.

6-H), 8.42 (s, -(CH3)2).

Formulation examples are explained below.

Example A (Dry powder for injection) Pyle (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-
2-oximino)acetamido)-3-(1-pyridiniummethyl)cef-3-em-4-carboxylate 500IItgLysine acetate
1891Rg cephalosporin antibiotic was blended with lysine acetate and filled into glass vials.

The headspace of the vial was purged with nitrogen and the combination was sealed with a crimp. The product was dissolved for administration by adding 2 d of Water for Injection.

Example B (Dry Powder for Injection) (6R,7R)-7-[(Z)-2-
(aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide)-3-(
1-Pyridinium methyl)cef-3-em-4-carboxylate monosodium salt is filled into a glass vial.

This filling is carried out under aseptic conditions under a sterile nitrogen atmosphere. The vial is closed using a rubber plate or stopper and held in place by an aluminum overseal to prevent gas exchange or microbial ingress. The product is reconstituted immediately prior to administration by dissolving in water for injection or other suitable sterile vehicle.

Example C (Twin pack for injection) (a) Sterility of 5004 (6R, 7R)-7-[(
Z)-2-(2-aminothiazol-4-yl)-2-
(2-Carboxyprop-2-oxyimino)acetamide)-3-(1-pyridiniummethyl)cef-3-em-4-carboxylate is aseptically packed into glass vials under sterile nitrogen. The vial is closed using a rubber plate or stopper held in place by an aluminum overseal, thereby preventing gas exchange or ingress of microorganisms.

(b) Prepare a 3.84% W/V sodium bicarbonate solution, clear by filtration and pack 2.15 m into clean ampoules. Pass carbon dioxide through the contents of each ampoule for 1 minute before sealing. Each ampoule is sterilized by autoclaving and examined for clarity.

(C) Reconstitute the cephalosporin antibiotic by dissolving it in 2, Od of the above sodium bicarbonate solution just before administration.

Example D (Dry powder for injection) (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-<1-carboxycyclobut-1 -oximino)acetamide) = 3-(1-pyridiniummethyl)ceph-3-em-4-carboxylate 5001 + 19 anhydrous sodium carbonate
The 47η cephalosporin antibiotic was combined with carbonate and filled into glass vials. The headspace of the vial was purged with nitrogen and the combination was sealed with a crimp. The product was dissolved for administration by addition of 2 m water for injection.

Example E (veterinary injection) Near one (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclobuto-1-oximino)acetamide) -3-(1-Pyridinium methyl)cef-3-em-4-carboxylate 10% W/V Aluminum distearate was dispersed in ethyl ocyanate and heated to 150°C for 1 hour with stirring, then allowed to cool to room temperature. Cooling. The sterile, ground antibiotic is added aseptically to the vehicle and purified in a high speed mixer. The product is aseptically packaged into injection vials and sealed with rubber seals or stoppers held in place by aluminum overseals.

Next, biological test results regarding the compounds of the present invention are shown.

Test compound A: (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide)-3-(1- Pyridinium ethyl) cef-3-em-4-carboxylate.

B: (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide) -3-(4
-rubamoyl-1-pyridiniummethyl)-cef-3
-Em-4-carboxylate, sodium salt.

C: (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyflobutol 1-oximino)acetamide)-3-(,
1-pyridinium methyl)-cef-3-emu 4-carboxylate.

D: (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclobudo-2-oximino)acetamide)-3-(
4-Carbamoyl-1-pyridiniummethyl)-ceph-
3-M-4-carboxylate, sodium salt.

E: (6R,7R)-7-[(Z)-2-(2-
aminothiazol-4-yl)-2-(1-carboxycycloprop-2-oximino)acetamide)-3
-('l-pyridiniummethyl)-cef-3-m-4
-Carboxylate, sodium salt.

F: (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclobent-1-yloxyimino)acetamide)-
3-(L-pyridiniummethyl)-cef-3-em-4
-Carboxylate, dihydrochloride.

Test method 1) Determination of the minimum inhibitory concentration (MIC) a) MIC against all microorganisms except Haemophilus influenzae:
CaS+tOne) was prepared in agar and poured into a vetri dish. Approximately 105 colony forming units/#li of the microorganisms listed in Table ■ on each plate! Inoculation was carried out using a multi-point inoculator from an inoculum containing . These microorganisms (however,
K, pneumoniae 1522 E) are all clinical isolates. After 18 hours of incubation at 37°C, the MIC in μ'J/d was determined as the minimum concentration for growth inhibition.

b) The MIC against Haemophilus influenzae was determined by the broth dilution method as in a) above.

However, brain heart infusion supplemented with Fildes and yeast extract was used as the medium.

2) Mouse protection test (ED5o) Groups of 5 mice each were challenged intraperitoneally with bacteria in 0.5-15% yeast suspension. 0.2 d of saline by mouth 1 hour and 5 hours after challenge with test compound dose.
It was administered subcutaneously twice as a solution.

The number of viable bacteria was counted on the fifth day after challenge, and the activity of the test compound was expressed as the average effective dose. The N. coli strain used is a cephalosporin-sensitive strain.

Table ■ A1.0 BO16 CO15 DO35 Fl, 5 3) β-lactamase stability (usually present in the 260-265 range) measured spectrophotometrically by tracking the ultraviolet absorption of the β-lactam ring; Its value is expressed relative to cephaloridine (which is assumed to have a stability value of 1 for each enzyme).

Results The test compound was p99. It was stable, showing a stability value of >30 for cephaloridine against 1 and TEM β-lactamase.

4) Toxicity a) LD5o values of compounds A and C Aqueous solutions of the sodium salts of each compound were freshly prepared in sodium bicarbonate. A 20% W/V solution of each compound [
It was administered intravenously to a group of RH arpino mice (body weight 16-22 g), and the LD5° value was measured. The results shown in the following Table rV(a) were obtained.

b) The test compound was administered at 500 mf to each group of 5 female CRH Alpino mice (body weight 18-22 g).
A single dose of l/Kg was administered intraperitoneally.

Observation continued for 14 days.

Test Compound F did not cause death in the group of mice.

This means that the compound has an LD5 of 500 m9/Kg or more.
Indicates that it has a value of 0. Compounds C, D and σE were not tested.

Claims (1)

[Scope of Claims] (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2
-Oximino)acetamide]-3-(1-pyridiniummethyl)cef-3-em-4-carboxylate or its nontoxic salt as an active ingredient, a pharmaceutical product for use in humans or animals as an antibiotic against bacterial infections. composition.