JPH0257073B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to intermediates of cephalosporin compounds with valuable antibiotic properties. The cephalosporin compounds herein are named in relation to "cephalosporin" according to "J. Amer. Chem. Soc.", Vol. 84, p. 3400 (1962);
The term "cephaem" refers to the basic cephaem structure with one double bond. Cephalosporin antibiotics are widely used in the treatment of diseases caused by pathogenic bacteria in humans and animals, and for the treatment of diseases caused by bacteria that are resistant to other antibiotics, such as penicillin compounds, and in the treatment of patients susceptible to penicillin. Particularly useful. In many cases, it is desirable to use cephalosporin antibiotics that exhibit activity against both Gram-positive and Gram-negative bacteria, and a significant amount of research has been devoted to the development of various types of broad-spectrum cephalosporin antibiotics. There is. That is, for example, the inventors' UK patent no.
1399086 (Japanese Unexamined Patent Publication No. 48-4487) describes 7β-(α
A new class of cephalosporin antibiotics containing a -etherified oxyimino)-acylamido group 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 Gram-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. In British Patent No. 1496757 (Japanese Unexamined Patent Publication No. 105689/1989) by the present inventors, the formula [In the formula, R is a thienyl group or a furyl group,
R ^A and R ^B can vary widely, for example, can form a C _1-4 alkyl group or together with the carbon atom to which they are attached a C _3-7 cycloalkylidene group, and m and n are m
and each 0 such that the sum of n is 0 or 1
Cephalosporin antibiotics containing a 7β-acylamido group have been described, and these compounds are syn isomers or at least 90%
is a syn and anti isomer mixture containing the syn isomer. 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, in British Patent No. 1522140 (Japanese Unexamined Patent Publication No. 51-91287) of the present inventors, the formula [In the formula, R ¹ represents a furyl group or a thienyl group, R ² represents a C ₁ -C ₄ alkyl group, a C ₃ -C ₇ cycloalkyl group, a furylmethyl group or a thienylmethyl group, and R ³ represents a hydrogen atom cephalosporin antibiotics (representing a carbamoyl, carboxy, carboxymethyl, sulfo or methyl group) are described, and these compounds are syn isomers or contain at least 90% syn isomers. Exists as a mixture of syn and anti 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. These compounds and others of similar structure have been developed in further attempts to find antibiotics with improved broad-spectrum antimicrobial activity and/or high activity against Gram-negative microorganisms. Such developments included changes in the 7β-acylamido group in the above formula as well as the introduction of specific groups at the 3-position of the cephalosporin molecule. That is, for example, Belgian Patent No. 852,427 (Japanese Unexamined Patent Publication No. 125190/1986) describes a cephalosporin antibiotic compound that is within the general scope of the present inventors' UK Patent No. 1,399,086. , where the above formula
The group R in (A) may be substituted by a variety of different organic groups, including the 2-aminothiazol-4-yl group, and the oxygen atom in the oximino group may itself be substituted by a fatty acid, which may itself be substituted, for example by carboxy. bonded to a group hydrocarbon group. 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. 836813 (Japanese Patent Application Publication No. 836813)
51-75066), the group R in the above formula (A) may be substituted with, for example, a 2-aminothiazol-4-yl group, and the oximino group is a hydroxyimino group or a blocked hydroxy group. Cephalosporin compounds are described that have imino 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. 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. 853545
125188), the 7β-acylamide side chain is mainly 2-(2-aminothiazol-4-yl)-2
A cephalosporin antibiotic is described which is a -(syn)-methoxyiminoacetamide group and in which the substituent in the 3-position is broadly defined as in Belgian Patent No. 836,813. 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. The inventors have discovered that 7β-
It has been discovered that by appropriate selection of a few specific groups at positions, cephalosporin compounds can be obtained that have particularly advantageous activity (detailed below) against a wide range of commonly encountered pathogenic microorganisms. The present invention is based on the general formula [In the formula, B is S or S→O, R ⁷ is an amino group or a protected amino group, and R ^8a
represents a hydrogen atom or a carboxyl-occluding functional group, or has a group of the formula -COOH and an associated anion A in place of the group -COO in the 4-position of the above formula. However, in the above formula, when B is S, R ^8a is a hydrogen atom, and R ⁷ is an amino group, the group at the 4-position in the above formula is COO ^- or
Not COOH. ] A cephalosporin compound or a salt thereof is provided. The compounds according to the invention are syn isomers. This syn isomeric form is radical with respect to the carboxamide group. defined by the arrangement of In this specification, thin configuration is structurally is shown as 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 (particularly hydrates) of the compound of formula (). It also includes within its scope salts of compounds of formula (). 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. That will be understood. Furthermore,
The compounds of formula () described above can also exist in another zwitterionic form, for example in which the 4-carboxyl group is protonated and the 7-carboxyl group in the side chain is deprotonated;
This alternative form is also included within the scope of the invention. The compounds according to the invention exhibit broad-spectrum antimicrobial activity. It has unusually high activity against Gram-negative microorganisms. 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 Gram-negative microorganisms. The compounds according to the invention are suitable for Pseudomonas (Pseudomonas).
-domonas) microorganisms such as Pseudomonas aeruginosa
unusually high activity against strains of Enterobacteriaceae (e.g. Escherichia coli, Klebsiella
Klebsiella pneumoniae, Salmonella typhimurium
typhimurium), Shigella sonnei (Shigella
sonnei), Enterobacter cloacae, Serratia marcescens, Providence spp., Proteus mirabilis and especially for example Proteus vulgaris and Proteus morgani morganii) and Haemophilus influenzae (indole-positive Proteus microorganisms such as
It was found to exhibit high activity against strains of influenzae. 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 generally 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. Thus, 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 () include alkali metal salts (e.g. sodium and potassium salts) and alkaline earth metal salts ( Inorganic base salts (e.g. calcium salts), amino acid salts (e.g. lysine salts and arginine salts), organic base salts (e.g. procaine salts, phenylethylbenzylamine salts, dibenzylethylenediamine salts,
ethanolamine salt, diethanolamine salt and N-methylglucosamine salt). Examples of other non-toxic salt derivatives include acid addition salts formed, for example, with hydrochloric, hydrobromic, sulfuric, 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 () can be used therapeutically because upon administration, such salts are rapidly distributed into the body. However, if insoluble salts of compound () are desired for use in a particular application, eg, in a depot 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 () in the processes described below. Examples of non-toxic metabolically unstable ester derivatives that can be produced by esterifying either or both carboxyl groups in the base compound of formula () include acyloxyalkyl esters, lower alkanoyloxymethyl esters, etc. or lower alkanoyloxyethyl esters such as acetoxymethyl ester, acetoxyethyl ester or pivaloyloxymethyl ester. Besides the above-mentioned ester derivatives, the present invention includes within its scope other physiologically acceptable equivalents of compounds of formula (), i.e. compounds of formula ( ) that is converted into the original antibiotic compound. More preferred compounds of the invention because of their high antibacterial activity are of the formula (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide]-
3-(1-pyridiniummethyl)-cef-3-em-4-carboxylate and its non-toxic salts (eg, sodium salt) and non-toxic metabolically unstable esters. Compounds of formula (b) significantly possess the general antimicrobial properties described above for compounds of general formula (). However, it may be emphasized 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. It is sufficiently distributed throughout the body of small rodents after subcutaneous injection to provide useful therapeutic levels. In primates, this compound gives 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 Gram-negative bacteria in mice have been successfully treated using this compound, and in particular Pseudomonas aeruginosa, an organism not normally susceptible to treatment with cephalosporin antibiotics.
Remarkable protection was obtained against the A. aeruginosa strain. This protection was comparable to treatment with aminoglycosides such as amikacin. Acute toxicity studies on this compound in mice gave LD ₅₀ values of greater than 1.0 g/Kg. No nephrotoxicity was observed in rats at a dose of 2.0 g/Kg. Another compound with properties not significantly different from the compound of formula (b) is (6R,7R)-7-[(Z)-2-(2-
aminothiazol-4-yl)-2-(1-carboxycyclobut-1-oximino)acetamide]-3-(1-pyridiniummethyl)-cef-3
-Em-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, compounds of the following formula () 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-pyridinium methyl)-cef-3-em-4-carboxylate, (6R,
7R)-7-[(Z)-2-(2-aminothiazole-
4-yl)-2-(1-carboxycycloprop-
1-oximino)acetamide]-3-(1-pyridiniummethyl)-cef-3-em-4-carboxylate, (6R,7R)-7-[(Z)-2-(2-aminothiazole-4- yl)-2-(1-carboxycyclopent-1-yloxyimino)acetamide]-3-(1-pyridiniummethyl)-cef-3
-M-4-carboxylate and (6R,
7R)-7-[(Z)-2-(2-aminothiazole-
4-yl)-2-(1-carboxycyclobut-1
-oximino)acetamide]-3-(4-carbamoyl-1-pyridiniummethyl)-cef-3
-Em-4-carboxylate. Compounds of formula () 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 R ⁴ has the above definition, B is >S or >S→O (α- or β-) and 2-
The dotted lines bridging the 3-, 3- and 4-positions indicate that the compound is Cef-2-M or Cef-3
A 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 inorganic acid such as methanesulfonic acid or toluene-p- or N-silyl derivatives or N-silyl derivatives with the formula -COOR ⁵ in the 4-position [formula R ⁵ is a hydrogen atom or a carboxyl-occluding functional group, such as an ester-forming aliphatic alcohol or is the residue of an aromatic aliphatic alcohol or an ester-forming phenol, silanol or stannol (the alcohol, phenol, silanol or stannol preferably having from 1 to 20 carbon atoms)
The corresponding compound having a group of in which R ^a and R ^b have the above definitions, R ⁶ represents a carboxyl-occluding group, for example a group as described for R ⁵ and R ⁷ is an amino group or a protected amino group. acylated with an acid or corresponding acylating agent, or (B)
formula [wherein R ^a , R ^b , R ⁷ , B and the dotted line have the above definitions, R ⁸ and R ^8a can independently represent hydrogen or a carboxyl occluding group, and X is a nucleophile. a substitutable residue such as an acetoxy group or a dichloroacetoxy group or a halogen atom such as chlorine, bromine or iodine] or a salt thereof with the formula with a pyridine compound in which R ⁴ has the above definition, followed by the following reaction in any suitable order, as required and/or desired in each case: Δ to the desired Δ ³ -isomer. ² - Conversion of isomers, reduction of compounds in which B is >S→O to produce compounds in which B is >S, non-toxic salts of carboxyl groups or non-toxic metabolically unstable Conversion to ester functionality and carboxyl blocking functional group and/or N-
There is provided a process for the preparation of an antibiotic compound of general formula () or a non-toxic salt or non-toxic metabolically labile ester thereof, having the above definitions, comprising carrying out either the removal of a protecting group. In the method (A), the starting material of formula () is preferably a compound in which B is >S and the dotted line represents a cef-3-em compound. One of the starting materials found to be particularly suitable for use in process (A) due to the high purity that can be produced is N-
(7-aminocef-3-em-3-ylmethyl)
Pyridinium-4-carboxylate dihydrochloride. As examples of acylating agents which can be used for the preparation of compounds of formula (), mention may be made, for example, of acid halides, especially acid chlorides or acid bromides. Such acylating agents can be prepared by reacting the acid () or a salt thereof with a halogenating agent such as phosphorus pentachloride, thionyl chloride or oxalyl chloride. Acylation with acid halides can be carried out in aqueous and non-aqueous reaction media, conveniently at temperatures of -50 to +50°C, preferably -20 to +30°C, optionally in the presence of acid binding agents. 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),
Mention may be made of amides (eg dimethylacetamide), nitriles (eg acetonitrile) or mixtures of two or more such solvents. 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 () can themselves be used as acylating agents in the preparation of compounds of formula (). Acylation using acids () is carried out using condensing agents such as N,
N'-dicyclohexylcarbodiimide or N
- in the presence of a carbodiimide, such as ethyl-N'-γ-dimethylaminopropylcarbodiimide, a carbonyl compound, such as carbonyldiimidazole, or an isoxazolium salt, such as N-ethyl-5-phenyl isoxazolium perchlorate. It is desirable to carry out the Acylation also forms other amides of acids of formula (), 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 sexual derivatives. Mixed anhydrides include phosphoric acid (e.g. phosphoric acid or phosphorous acid), sulfuric acid or aliphatic or aromatic sulfonic acids (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 carried out in anhydrous reaction media such as methylene chloride, tetrahydrofuran,
Preferably it is carried out in 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 () and their corresponding acylating agents can optionally be produced and used in the form of their acid addition salts. Thus, for example, it is convenient to use acid chlorides as their hydrochlorides and acid bromides as their hydrobromides. The pyridine compound of formula () can act as a nucleophilic reagent to substitute various substituents X from the cephalosporin of formula (). To some extent, the ease of substitution is related to the pKa of the acid HX from 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. or the ease of substitution is to some extent the expression ()
It is also related to the nature of the substituent R ⁴ in the compound. The replacement of X by a pyridine compound of formula () 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 () in which the substituent X is, for example, a halogen atom or an acyloxy group as described below. Compounds of formula () in which X is an acetoxy group are convenient starting materials for use in nucleophilic substitution reactions with pyridine compounds of formula ().
Examples of other starting materials of this type include, for example, in formula () where X is substituted acetic acid, such as chloroacetic acid;
Mention may be made of compounds which are the residues of dichloroacetic acid and trifluoroacetic acid. The substitution reaction for compounds having 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. 1132621 (Patent No. 757847) and British Patent No. 1171603 (Patent No.
No. 634584) are detailed in each specification. Substituent X may also be derived from formic acid, haloformic acid (eg chloroformic acid) or carbamic acid. When using a compound in which X represents an acetoxy group or a substituted acetoxy group in formula (), it is generally desirable that the group R ⁸ in formula () is a hydrogen atom and B represents >S. In this case, the reaction is preferably carried out at a pH of 5 to 8, particularly preferably 5.5 to 7.
It is advantageous to carry out in an aqueous medium at PH. The method using a compound in which X is the residue of a substituted acetic acid is described in British Patent No. 1,241,657.
(Patent No. 833410). When using a compound in which X is an acetoxy group in formula (), the reaction is carried out at 30° to 110°C, preferably
It is convenient to carry out at a temperature of 50° to 80°C. Compounds of formula () 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 (). This kind of expression ()
When using the compound B is>S→O and
R ⁸ can represent a carboxyl occluding group. The reaction is preferably carried out in one or more organic solvents, preferably polar ones, such as ethers (such as dioxane or tetrahydrofuran), esters (such as ethyl acetate), amides (such as formamide and N,
It is convenient to carry out the reaction in a non-aqueous medium consisting of N-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 (Patent No.
No. 874185) is detailed in the specification. The reaction medium must neither be extremely acidic nor extremely basic. In the case of reactions carried out on compounds of formula () in which R ⁸ and R ^8a are carboxyl occluding groups, the 3-pyridinium methyl product is formed as the corresponding halide salt, which can be optionally treated one or more times. A salt having a desired anion can be obtained by subjecting it to an ion exchange reaction. When using a compound in which X is the above-mentioned halogen atom in formula (), the reaction temperature is -10° to +50°C,
It is convenient to carry out preferably at a temperature of +10 to +30°C. The reaction product can be produced by e.g. recrystallization, iontophoresis,
reaction mixtures that may contain e.g. unreacted cephalosporin starting material and other materials by a number of methods including column chromatography and the use of ion exchangers (e.g. by chromatography on ion exchange resins) or macroreticular resins. can be separated from The △ ² -cephalosporin ester derivative obtained according to the method of the present invention is, for example, the △ ²
− Corresponding △ ³ − by treating the ester with a base such as pyridine or triethylamine.
Can be converted into derivatives. 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→O, 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. Reaction is -20° to +50
It can be carried out at a temperature of °C. Metabolically unstable ester derivatives of compounds of formula () may be prepared by preparing a compound of formula () or a salt or protected derivative thereof, conveniently in an inert organic solvent such as dimethylformamide or acetone, with an acyloxyalkyl halide (e.g. iodide). ), and then, if necessary, removing the protecting group. Base salts of compounds of formula () may be produced by reacting acids of formula () 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 () or a metabolically unstable ester derivative thereof with a suitable acid. When the compound of formula () 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 () according to the invention, syn-isomers and the corresponding anti-isomers which 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 () and the corresponding acid halides and acid anhydrides in the form of mixtures. The acid of the formula () (provided that R ^a and R ^b do not form a cyclopropylidene group together with the carbon atom to which they are bonded) is of the formula A compound ^of the ^general formula etherification by reaction with a compound in which R ^a and R ^b and R ⁶ have the above definitions and T is a halogen such as chloro, bromo or mode, a sulfate or a sulfonate such as tosylate; It can be produced by removing the carboxyl occluding group R ⁹ . 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,N
- Preferably carried out in a disubstituted amide (for example 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 (). The base should be used in an amount sufficient to rapidly neutralize the acid in question. Acids of general formula () can also be expressed as A compound of the formula [wherein R ⁷ and R ⁹ have the above definitions] [In the formula, R ^a , R ^b and R ⁶ have the above definitions]
, followed by removal of the carboxyl occluding group R ⁹ and optionally separation of the syn and anti isomers. The last-mentioned reaction is particularly applicable to the preparation of acids of formula () in which R ^a and R ^b together with the carbon atoms to which they are attached form a cyclopropylidene group. In this case, the related compounds of formula () can be prepared in a conventional manner, for example by the synthesis method described in Belgian Patent No. 866,422 for the preparation of tertiary-butyl 1-aminooxycyclopropane carboxylate. Acids of formula () 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 () is a halogen (i.e. chlorine, bromine or iodine) atom, the cef-3-em starting compound can be converted to 7β-protected amino-3-methyl cef-3, for example, in a manner analogous to method (A) above.
Halogenating the 3-m-4-carboxylic acid ester 1β-oxide and removing its 7β-protecting group,
They may be prepared in a conventional manner by 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 a British patent no.
It is described in the specification of No. 1326531 (Patent No. 874185). The corresponding cef-2-em compounds are prepared by reacting 3-methylcef-2-em compounds with N-bromosuccinimide according to the dimensions described in Dutch Patent Application No. 6902013 to obtain the corresponding 3-bromomethylcef-2-em compounds. M-compounds can be obtained. When X is an acetoxy group in formula (),
Such starting materials can be prepared, for example, by acylating 7-aminocephalosporanic acid in a manner analogous to method (A) above. formula()
Compounds in which X represents another acyloxy group are, for example, as described in British Patent No. 1474519 (Japanese Unexamined Patent Publication No. 1974-35395) and British Patent No. 1531212 (Japanese Unexamined Patent Publication No. 51-76489). Appropriate 3-
It can be produced by acylating the corresponding 3-hydroxymethyl compound, which can be produced by hydrolysis of an acetoxymethyl compound. Also, the starting material of formula () is British Patent No. 1028563
As described in 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 () is to prepare the corresponding protected 7β using e.g. PCl ₅ in a conventional manner.
- consists of deprotecting the amino compound. 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 it may be necessary to protect the NH ₂ group of the aminothiazolyl moiety, for example by tritylation, acylation (eg chloroacetylation), protonation or other conventional methods. 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 trifluoroacetic acid. removed using acetic 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. sell. The carboxy-occluding functional group used in the preparation of the compound of formula () or the necessary starting materials is preferably a group that can be easily cleaved at an appropriate step of the reaction process, conveniently at the final step. However, in some cases non-toxic metabolically labile groups such as acyloxymethyl or acyloxyethyl groups (e.g. acetoxymethyl or acetoxyethyl or pivaloyloxymethyl) may be used to obtain suitable ester derivatives of compounds of formula (). It is convenient to use carboxyl-occluding functional groups and retain these in the final product. Suitable carboxy blocking groups are well known in the art, and a list of representative blocked carboxyl groups is described in British Patent No. 1399086 (Japanese Patent Publication No. 48-4487). There is.
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 lower haloalkoxycarbonyl groups (for example, 2,2,2-trichloroethoxycarbonyl). The carboxyl-occluding functional 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. pharmaceutical compositions containing antibiotic compounds according to the invention adapted for 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 suspending, 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 invention can also be prepared as suppositories, containing suppository bases such as cocoa butter or other glycerides. Veterinary pharmaceutical compositions may also be formulated, for example, as either long-acting or rapid-release intramammary formulations. The composition may be 0.1% or more depending on the administration method, e.g. 0.1% or more.
May contain ~99% active substance. When the composition consists of dosage units, each unit preferably contains from 50 to 1500 mg of active ingredient. The dosage used for the treatment of adults is preferably 500 to 6000 mg per day, depending on the route and frequency of administration. For example, in the treatment of adults, if administered intravenously or intramuscularly, 1
1000-3000 mg per day will usually 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 penicillin or other antibiotics such as cephalosporins. Next, the present invention will be explained by examples. All temperatures are in °C. "Petrol" means petroleum ether (boiling point 40-60°). Proton magnetic resonance (pmr) spectrum is 100MHz
was measured. Each integral matches the assignment,
The coupling constant J is expressed in Hz, and each sign is s = single line, d = doublet, dd = overlapping doublet,
m=multiplet and ABq=AB quartet were not measured. Preparation 1 Ethyl (Z)-2-(2-aminothiazol-4-yl)-2-(hydroxyimino)acetate A solution of ethyl acetoacetate (292 g) in glacial acetic acid (296 ml) was stirred and cooled on ice; Add to this sodium nitrite (180 g) in water (400 ml) at a rate such that the reaction temperature is maintained below 10°C.
A solution of was added. Stirring and cooling were continued for approximately 30 minutes, then a solution of potassium chloride (160g) in water (800ml) 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 petrol and then dried in vacuo over potassium hydroxide to give ethyl (Z)-2-(hydroxyimino)-3-oxobutyrate (309 g).
I got it. Ethyl (Z)-2- in dichloromethane (400 ml)
A stirred and ice-cooled solution of (hydroxyimino)-3-oxobutyrate (150 g) was treated in portions 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 (177g) was dissolved in ethanol (500ml) and dimethylaniline (77ml) and thiourea (42g) 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, melting point 188° (decomposed)). Production method 2 Ethyl (Z)-2-hydroxyimino-2
-(2-tritylaminothiazole-4-
acetate hydrochloride A solution of the product of Preparation 1 (12.91 g) in dimethylformamide (28 ml) containing triethylamine (8.4 ml) was stirred and cooled (-30°),
Trityl chloride (16.75 g) was added dropwise to this over a period of 2 hours. Leave the mixture for 1 hour
The mixture was allowed to warm to 15°, stirred for a further 2 hours and then partitioned between water (500ml) and ethyl acetate (500ml). The organic phase was separated, washed with water (2x500ml) and stirred with 1NHCl (500ml). Collect the precipitate,
Washing successively with water (100ml), ethyl acetate (200ml) and ether (200ml) followed by drying in vacuo gave the title compound as a white solid (16.4g, mp 184-186° (dec)). Production method 3 Ethyl (Z)-2-(2-tert-butoxycarbonylprop-2-oximino)-2-
(2-trityl-aminothiazole-4-
A solution of the product of Preparation 2 (49.4 g) in dimethyl sulfoxide (200 ml) was stirred under nitrogen and added with tert-butyl 2-bromo-2-methylpropiodine in dimethyl sulfoxide (25 ml). nate (24.5g) and potassium carbonate (34.6
g) was added and the mixture was stirred for 6 hours at room temperature. The mixture was poured into water (2), stirred for 10 minutes and filtered. The solid was washed with water and dissolved in ethyl acetate (600ml). The solution was washed successively with water, 2N hydrochloric acid, water and saturated brine, dried and evaporated. The residue is converted into petroleum ether (boiling point 60~80°)
The title compound [34 g, melting point 123.5] was recrystallized from
~125°] was obtained. Production method 4 (Z)-2-(2-tert-butoxycarbonylprop-2-oximino)-2-(2-
Tritylaminothiazol-4-yl)acetic acid The product (2 g) of Production Method 3 was added to methanol (20 ml).
To this add 2N sodium hydroxide (3.3
ml) was added. The mixture was refluxed for 1.5 hours and then concentrated. The residue was taken up in a mixture of water (50ml), 2N hydrochloric acid (7ml) and ethyl acetate (50ml). The organic phase was separated and the aqueous phase was extracted with ethyl acetate. The organic solutions were combined, washed successively with water and saturated brine, dried and evaporated. The residue was recrystallized from a mixture of carbon tetrachloride and petrol to give the title compound [1 g, melting point 152-156° (decomposed)].
I got it. Process 5 Ethyl (Z)-2-(2-tritylaminothiazol-4-yl)-2-(1-tert-butoxy-carbonylcyclobut-1-oximino)acetate The product of Process 2 (55.8 g) Stirred with potassium carbonate (finely ground, 31.2 g) in dimethyl sulfoxide (400 ml) at room temperature under nitrogen. 30
After a few minutes, tertiary butyl 1-bromocyclobutanecarboxylate (29.2 g) was added. After 8 hours, additional potassium carbonate (31.2 g) was added. Next 3
More potassium carbonate (6 x 16 g portions) was added during the next day and after 3 days more tertiary butyl 1-bromocyclobutane carboxylate (3.45 g) was added. After a total of 4 days the mixture was poured into ice-water (approximately 3) and the solids were collected by filtration and washed thoroughly with water and petrol. The solid was dissolved in ethyl acetate and the solution was washed with brine (twice),
Drying over magnesium sulfate and evaporation gave a foam. This foam was dissolved in ethyl acetate-petrol (1:2) and passed through silica gel (500 g). Evaporation gave the title compound (60g) as a foam. νmax
( _CHBr3 ) 3400 (NH) and 1730 cm ^-1 (ester). Production method 6 (Z)-2-(1-tertiary-butoxycarbonylcyclobut-1-oximino)-2-
(2-trityl-aminothiazole-4-
A mixture of the product of Preparation 5 (3.2g) and potassium carbonate (1.65g) was refluxed in methanol (180ml) and water (20ml) 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, to which was added 2NHCl (12.2
ml) 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 (2.3g). λmax (ethanol) 265nm (E1% 1cm243). Production method 7 (Z)-2-(1-tertiary butoxycarbonylcycloprop-1-oximino)-2
-(2-tritylaminothiazole-4-
A solution of hydrazine hydrate (0.20 g) in methanol (0.4 ml) was dissolved in dichloromethane (7 ml).
1-tert-butoxycarbonylcycloprop-1-oxyphthalimide (0.61 g,
(prepared as described in Belgian Patent No. 866,422). The mixture was stirred for 1 hour at room temperature and then treated with 5N aqueous ammonia solution (7ml). The organic phase was separated and the aqueous phase was extracted with dichloromethane. The organic solutions were combined, washed with water, dried and evaporated. The oily residue (0.30g) was dissolved in a mixture of ether (5ml) and ethyl acetate (5ml). 2-tritylaminothiazol-4-ylglyoxylic acid (0.73
g, prepared as described in Belgian Patent No. 864,828). The mixture was stirred overnight at room temperature and then filtered. The solid was washed with a small amount of ether and dried in vacuo to give the title compound (0.5 g, mp 156.8-157.2°). νmax
( _CHBr3 ) 2300-3500 (O-H, N-H), 1750
(tertiary butyl ester), 1690 cm ^-1 (acid). Production method 8 Ethyl (Z)-2-(1-tert-butoxycarbonylcyclopent-1-yloxyimino)-2-(2-tritylaminothiazol-4-yl)acetate The product of production method 2 (10 g) Tertiary-butyl 2-bromo-cyclopentanecarboxylate (7 g) in dimethyl sulfoxide (40 ml) containing potassium carbonate (10 g) for 21 hours at 21° under nitrogen.
and stirred together. The mixture was poured into ice-water (500ml) and the gray solid was filtered off, washed with water and air dried. This solid was recrystallized from methanol (500ml) to give the title compound (11.7g, melting point 179-180°). νmax ( _CHBr3 ) 3410 (NH), 1735 (ester), 1275 (ester) and 755 cm ^-1 (phenyl). Production method 9 (Z)-2-(1-tertiary butoxycarbonylcyclopent-1-yloxyimino)
-2-(2-Trityl-aminothiazol-4-yl)acetic acid The product of Preparation 8 (625 mg) was refluxed for 7 hours with 2N sodium hydroxide solution in methanol (0.5 ml) and water (1 ml). The mixture was allowed to cool overnight. After dilution with water, orthophosphoric acid was added to adjust the solution to pH 2. The precipitate was extracted with ether and the combined extracts were washed with brine.
After drying over magnesium sulfate, the solvent was evaporated to give a gum (493 mg). Recrystallization from diisopropyl ether gave the title compound (356 mg, melting point 171-173°).
I got it. νmax( _CHBr3 )2500−3500(OH and
NH), 1755 (ester), 1692 (acid) and 755 and 770 cm ^-1 (phenyl). Preparation 10 (6R,7R)-7-amino-3-(1-pyridiniummethyl)cef-3-em-4-carboxylic acid dihydrochloride (a) (6R,7R) in dichloromethane (30 ml)
7R)-7-(2-thienylacetamide)-3-
A stirred suspension of (1-pyridinium-methyl)cef-3-em-4-carboxylate (4.15g) was treated with N,N-dimethylaniline (5.09ml) and chlorotrimethylsilane (2.52ml). The mixture was stirred for 1 hour at 30-35°, cooled to -28°, and phosphorus pentachloride (4.16 g)
After stirring for a further 1 hour at -25° to -30° it was poured into a stirred, cooled (-20°) solution of dichloromethane (20 ml) and butane-1,3-diol (8.1 ml). The solution was left to reach a temperature of 0° for 30 minutes and the precipitated solid (A) was filtered off, washed with dichloromethane and dried in vacuo. Add it to methanol (17.5
ml), stirred and diluted with dichloromethane (87.5ml) and the precipitated solid was filtered, washed with dichloromethane and dried in vacuo to give the title compound as a white solid (3.2g). . λmax (PH6 buffer) 258nm (E
1% 1cm318), τ( _D2O ) values are 0.95, 1.32 and 1.84 (pyridinium proton), 4.10-4.46 (ABq, J16Hz, 3
-CH ₂ -), 4.56 (d, J5Hz, 7-H), 4.70 (d,
J5Hz, 6-H), 6.14~6.50 (ABq, J17Hz, C ₂
-H). (b) The solid (A) produced in step (a) above was dissolved in 1N hydrochloric acid (25 ml). Isopropanol (95
ml) to precipitate the crystalline title compound as the dihydrate (4.95 g). The τ(D ₂ O) value is
1.02, 1.36 and 1.87 (pyridinium protons),
4.2 + 4.55 (ABq, J = 14Hz, 3-CH ₂ -), 4.62
(d, J=5Hz, C ₇ −H), 47.4(d, J=5Hz,
C ₆ −H), 6.19+6.38 (ABq, J=18Hz, C ₂ −
H). The water content according to Karl Fitscher method is 9.4%. Example 1 a Tertiary butyl (6R,7R)-3-acetoxymethyl-7-[(Z)-2-(2-tert-butoxycarbonylprop-2-oximino)-2-
(2-tritylaminothiazol-4-yl)
Acetamide] cef-3-em-4-carboxylate The product of Preparation 4 (572 mg) and tertiary butyl (6R,
7R)-3-acetoxymethyl-7-aminocef-
A stirred solution of 3-em-4-carboxylate (328 mg) was cooled to 0° and to this was added 1-hydroxybenzotriazole (150 mg) followed by dicyclohexylcarbodiimide (225 mg). The mixture was warmed to warm, 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 liquid and washings were diluted with water (50ml) and extracted with ethyl acetate. The combined organic extracts were washed successively with water, 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 (533mg). A portion was recrystallized from diisopropyl ether. Melting point 103-113° (decomposed), [α] ²⁰ _D +8.5° ( c , 1.0, DMSO). b (6R,7R)-3-acetoxymethyl-7-
[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide] Cef-3-M-4-
Carboxylic Acid To a solution of the product from a above (2.4 g) in anisole (18 ml) at 0° trifluoroacetic acid (18 ml) 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 PH1.5 under ethyl acetate, saturated with sodium chloride and extracted with ethyl acetate. The organic extracts were combined, washed with saturated brine, dried and evaporated.
The residue was dissolved in a hot 50% formic acid aqueous solution (20ml) and allowed to stand for 2 hours. The mixture was diluted with water (50ml) and filtered. The liquid was concentrated. The residue was taken up in water (50ml), filtered again and lyophilized to give the title compound (920mg). λmax (PH6 buffer) 236nm (E1% 1cm250), λinf255nm (E1% 1cm235), 296nm (E1% 1cm103), [α] ²⁰ _D + 20.0° ( c 1.0, DMSO). c (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide]-
3-(1-pyridiniummethyl)-cef-3-em-4-carboxylate monosodium salt Pyridine (2 ml) and the product of b above (1.8 g) at 80° were dissolved in sodium iodide in water (2.2 ml). (7.12 g) into a stirred solution. The solution was stirred for 1 hour at 80°C, cooled and diluted with 100 ml of water.
diluted to The pH of the solution was adjusted to 6.0 with 2N sodium hydroxide solution and the solution was concentrated to remove pyridine. The aqueous residue was diluted to 100 ml with water, to which methyl isobutyl ketone (2 drops) was added and the solution acidified to PH1 with 2N hydrochloric acid.
The mixture was filtered and the solids were washed with a little water. The liquid and washings 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 ml and applied to a 500 g column of Amberlite XAD-2 resin using first water and then 20% ethanol in water as the eluent. The product containing fractions were concentrated and lyophilized to give the title compound (0.56g). λmax (PH6 buffer), 253.5nm (E1% 1cm307), λinf282nm (E1% 1cm159), 260nm (E1% 1cm295), [α] ²⁰ _D +24.5゜ ( c 1.0,
DMSO). Example 2 (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 monosodium salt Water containing sufficient sodium bicarbonate (0.7
To a stirred solution of the product of Example 1b) (0.59 g) in ml) was added isonicotinamide (0.56 g) to a final pH of 6.5. Sodium iodide (2.1g)
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 1c to yield the title compound (0.09g).
I got it. λmax (PH6 buffer) 257.5nm (E1% 1cm276), λinf291.5nm (E1% 1cm125), τ (D ₂ O) value is 0.92, 1.70 (4H, pyridinium proton), 3.10
(1H, aminothiazole-5-H), 4.34, 4.64
(2H, ABq, 3-CH ₂ -), 8.54 (6H, -CMe ₂ -)
Contains. Example 3 a Tertiary butyl (6R,7R)-3-acetoxymethyl-7-[(Z)-2-(1-tert-butoxy-carbonylcyclobut-1-oximino)
-2-(2-tritylaminothiazole-4-
Product of Preparation 6 (24.2 g) and tertiary butyl (6R,7R)-3-acetoxymethyl-7-aminocef- in dimethylformamide (300 ml) 3-M-4-carboxylate (13.6
The stirred solution of g) was cooled to 0° and treated with 1. hydroxybenzothiazole monohydrate (4.5 g) followed by dicyclohexylcarbodiimide (6.4 g) and the product was prepared essentially as described in Example 1a. The title compound (12.8 g), melting point 113.5 ~
116.5° (decomposition) was obtained. [α] ²⁰ _D +15.0゜( c 1.0,
DMSO). b(6R,7R)-3-acetoxymethyl-7-
[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclobut-1-oximino)acetamide] Cef-3-M-
4-Carboxylic acid Trifluoroacetic acid (100 ml) was added to a mixture of the product of a) above (12.5 g) and anisole (5 ml) at 0°. The mixture is substantially similar to Example 1b)
Treatment as described in gave the title compound (4g). λmax (PH6 buffer) 246nm (E1% 1cm
264), λinf295nm (E1% 1cm118), [α] ²⁰ _D +27.3゜( c
1.0, DMSO). c(6R,7R)-7-[(Z)-2-(aminothiazol-4-yl)-2-(1-carboxycyclobuto-1-oximino)acetamide]-3
-(1-pyridiniummethyl)-cef-3-em-4-carboxylate monosodium salt Iodination of pyridine (4.1 ml) and the product of b) above (3.75 g) in water (4.5 ml) at 80°C. Addition to a stirred solution of sodium (14.6g) and isolation of the product essentially as described in Example 1c) gave the title compound (1.3g). λmax(PH6
buffer) 252.5nm (E1% 1cm310), λinf291nm
(E1% 1cm139), [α] ²⁰ _D +43.5° ( c 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 Water containing sufficient sodium bicarbonate (1.3
Isonicotinamide ((1.22 g) was added to a stirred solution of the product of Example 3b) (1.08 g) in ml) to a final pH of 6.5. Sodium iodide (4 g)
was added and the mixture was stirred for 1 hour at 80°C.
Sodium bicarbonate was added occasionally to maintain a pH of ~6.5. The product was isolated essentially as described in Example 1c) to give the title compound (0.16g). [α] ²⁰ _D -18゜ ( c 1.08, H ₂ O), λmax (PH6 buffer) 256nm (E1% 1cm298), λinf294nm (E1% 1cm
135). Example 5 a Tertiary butyl (6R,7R)-3-acetoxymethyl-7-[(Z)-2-(2-tritylaminothiazol-4-yl)-2-(1-tertiary butoxycarbonyl Cycloprop-1-oxyimino)acetamide]cef-3-em-4-carboxylate Preparation of Preparation 7 from 1-hydroxybenzotriazole monohydrate (0.12 g) and dicyclohexylcarbodiimide (0.16 g) in tetrahydrofuran (6 ml) (0.34g) and tertiary butyl (6R,7R)-3-acetoxymethyl-7-
Added to a stirred solution of aminocef-3-em-4-carboxylate (0.25g). The mixture was stirred overnight at room temperature and then filtered. The 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. Concentrate the eluate to warm (0.44g)
This was recrystallized from diisopropyl ether (15 ml) to obtain the title compound (0.29 g). Melting point 115-119°, [α] ²⁰ _D ( c 1.0, DMSO) +13°. b (6R,7R)-3-acetoxymethyl-7-
[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycycloprop-1-
Oximino)acetamido]cef-3-em-4-carboxylic hydrochloride At 10° concentrated hydrochloric acid (0.6 ml) was added to a stirred solution of the product from a) above (1.92 g) in formic acid (7.5 ml). Ta. The mixture was stirred for 1.25 hours at room temperature and then filtered. The liquid was added to diisopropyl ether (300ml) and the mixture was stirred for 1.5 hours.
The solid was removed, washed with diisopropyl ether and diethyl ether and dried in vacuo to give the title compound (1.16g). [α] ²⁰ _D
( c 1.0, DMSO) +35°, λmax (PH6 buffer)
239nm (E1% 1cm300). c (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycycloprop-1-oximino)acetamide]-3-(1-pyridinium Methyl) cef-3
-Em-4-carboxylate sodium salt A mixture of the product from b above (0.56 g), sodium bicarbonate (0.17 g) and water (0.5 ml)
Warmed to 50°. Further sodium bicarbonate (0.09g) was added followed by pyridine (0.2ml). Warm the solution to 80° and add sodium iodide (2
g) was added. The solution was stirred for 40 minutes at 80°, cooled and diluted with acetone (50ml). The mixture was filtered and the solid was washed with acetone and ether to give a solid. This solid was dissolved in water (20ml) 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 (5 g) and passed through a pad of neutral alumina (10 g). 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 g) was dissolved in a small amount of water and 50 g of Amberlite
-2 resin column. The product containing fractions were concentrated and the residue was triturated with acetone to give the title compound (0.06g). [α] ²³ _D
0゜±1.5゜ ( c 0.1, water), λmax (PH6 buffer)
254nm (E1% 1cm340), λlnf296nm (E1% 1cm125). Example 6 (6R,7R)-7-[(Z)-2-aminothiazol-4-yl)-2-(1-carboxycyclopent-1-yl-oximino)-acetamide]-3-(1- Pyridinium methyl) Cef-3
-Em-4-carboxylate dihydrochloride Phosphorus pentachloride (0.46 g) was dissolved in methylene chloride (20 ml) at ambient temperature and the solution was cooled to 10° and the product of Preparation 9 (1.095 g) Added in the episode. The mixture was warmed to -5° and stirred for 30 minutes. The solution was cooled to -10° and added with triethylamine (0.61 ml) followed by water (6.7 ml) with vigorous stirring so that the temperature was not above 0° but the water did not freeze.
added. The two-phase mixture was stirred for 3 minutes and transferred to a tap. The lower phase was pre-cooled to -20° with N,N- containing triethylamine (1.4 ml).
Product of Preparation 10a (0.76
g) to the vigorously stirred suspension, the addition being such that the temperature did not rise above -10°. The mixture was stirred at -5 to -10° for 45 minutes and then allowed to warm to 21° over 1 hour. Methanol (0.3ml) was added and the methylene chloride was evaporated in vacuo at a bath temperature of 30°. The residue was dissolved in ethyl acetate (30ml) and water (30ml).
and a small amount of sodium chloride was added. The organic layer was further washed with water (2 x 30
ml). Combine the washings and additional sodium chloride and add ethyl acetate (20ml).
and the organic layers were combined and dried over magnesium sulfate. Evaporation gave a foam (1.79g) which was triturated with diisopropyl ether to give a solid (1.35g). Most of this solid (1.2g) was dissolved in formic acid (5ml) and concentrated hydrochloric acid (0.38ml) was added with vigorous stirring. After standing for 1 hour at 21°, the suspension was filtered and the residue was leached with a little formic acid. The combined liquids were concentrated by evaporation and the residue was triturated with acetone to give the title compound (374mg). [α] _D +8.6° ( c 1.02, H ₂ O) λmax (PH6 buffer) 255 nm (E1% 1cm289), λinf295 (E1% 1cm273), λinf280 (E1% 1cm158). Example 7 (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide]-3
-(1-pyridiniummethyl)-cef-3-em-4-carboxysodium salt (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2- Carboxyprop-2-oximino)acetamide]-3-(1
-Pyridinium methyl)-Cef-3-M-4-
The carboxylate (2.5g) was dissolved in water and the solution was treated with sodium 2-ethylhexanoate (1.52g) in methanol (8ml). The mixture was added to stirred acetone for 15 minutes and the resulting suspension was filtered, washed and dried to give the title compound (2.5g). [α] ²³゜_D 0゜( c 1.0
，
H ₂ O), λmax (PH6 phosphate), 255 (E1% 1cm
327, ε18630), λinf240 (E1% 1cm305, ε17370) and 280 (E1% 1cm172, ε9800), νmax (Nujiol) 1780cm ^-1 (β-lactam), actual value of sodium: 4.5%, C ₂₂ H ₂₁ O ₇ Calculated value as N ₆ S ₂ N _a :
4.04%. Example 8 a Diphenylmethyl (1S, 6R, 7R)-7-
[(Z)-2-(2-tertiary butoxycarbonylprop-2-oximino)-2-(2-tritylaminothiazol-4-yl)-acetamide]
-3-Bromomethylcef-3-em-1-oxide-4-carboxylate Phosphorus pentachloride (0.75g) was suspended in methylene chloride (20ml) with stirring. The mixture was cooled to -10° and the product of Preparation 4 (2.0 g) was added. Stirring was continued for 10 minutes at -5 to -10°. Triethylamine (0.88 ml) in methylene dichloride (5 ml) was added at -10°, then after 5 minutes methylene dichloride (30 ml) containing triethylamine (0.42 ml) was washed with methylene dichloride (5 ml).
diphenylmethyl (1S, 6R,
7R)-7-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 (50ml).
Separate the organic layer and add dilute hydrochloric acid solution (1N, 3 x 30ml)
and brine (2 x 30ml) and evaporated in vacuo to give a foam. The foam was taken up in ethyl acetate (approximately 10ml) and treated with diisopropyl ether (100ml). 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 τ( _CDCl3 ) value is 3.11
(s, -C H Ph ₂ ), 3.37 (s, thiazol-5-yl proton) 3.88 (dd, J9Hz and 5Hz, 7-
H), 5.22 + 6.02 (ABq-3CH ₂ ), 5.49 (d, 5Hz,
(6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2- oxyimino)acetamide]-
3-(1-pyridiniummethyl)-cef-3-em-4-carboxylate The product from a above (1 g) was dissolved in acetone (22 ml) and stirred at room temperature. Pyridine (0.08ml)
was added and the mixture was stirred for 3 hours at room temperature. Further pyridine (0.72ml) was added and the mixture was left at room temperature overnight. The mixture was poured into stirred diethyl ether (75 ml) and the precipitated solid was collected by filtration, washed with ether and concentrated in vacuo.
Dry at 40°. This solid (0.8g) was redissolved in acetone (22ml) at -10°. Potassium iodide (0.7g) was added followed by acetyl chloride (0.17ml). The mixture was stirred at -10° for 20 minutes and further potassium iodide (0.7g) and acetyl chloride (0.17ml) were added. After stirring for a further 20 min at −10°, the mixture was dissolved in sodium metabisulfite (0.6
g) was added to the solution. The product was extracted with methylene dichloride (2 x 50ml) and the extracts were washed with brine, dried over magnesium sulphate and evaporated under reduced pressure to give a foam. This was dissolved in formic acid (6.5ml) and left at room temperature for 15 minutes. Concentrated hydrochloric acid (0.25ml) was added and the mixture was left for a further 1.25 hours. The solid precipitate was filtered and washed with a small amount of formic acid. The solution and washing solution were combined and mixed with water (10ml) and acetonitrile (5ml).
ml) into ethyl acetate (5 ml) and diethyl ether (5 ml). More water was added until two distinct layers were obtained. Drain the lower layer and add Amberlite LA2 (7 ml) and acetic acid (0.7 ml).
ml) containing diethyl ether (14 ml). Separate the aqueous layer again and zeolite
It was applied to a column of 225SRC15 (H ⁺ form 15ml).
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. λmax (PH6 phosphate) at 255.5 nm (E1% 1 cm374), λinf at 238 (E1% 1 cm340) and 290 nm (E1% 1 cm160). Example 9 a (6R,7R)-7-[(Z)-2-(2-triphenylmethylaminothiazol-4-yl)-
2-(2-tert-butoxycarbonylprop-
2-oximino)acetamide]-3-(1-
Pyridinium-methyl)-cef-3-em-4
-Carboxylate To a stirred solution of phosphorus pentachloride (1.38 g) in methylene chloride (60 ml) (cooled to -10°) 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 (1.33g) was added followed by water (20ml). The mixture was stirred at 0° for 3 min and the lower phase was dissolved in N,N-dimethylacetamide (30 ml)/acetonitrile (30 ml) containing triethylamine (3.03 g).
ml) of the product of process 10(a) (2.19
g) to the stirred suspension (cooled to -10°) over 10 minutes. The mixture was stirred at -10° to -5° for 45 minutes and then for 1 hour without cooling. Methanol (1 ml) was added. Methylene chloride was removed by evaporation under reduced pressure. The residual solution was added to water (300ml) with stirring to precipitate the title compound (4.89g). The τ (CDCl ₃ ) value is 2.78 (s, −
[C ₆ H ₅ ] ₃ ), 3.37 (s,-thiazole proton),
0.35, 1.80, 2.12 (pyridinium proton), 4.18
(m, -7-H), 4.95 (6-H), 8.66 (s, -3rd
butyl), 8.50 (s, -C( _CH3 ) ₂ ). b (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide]-
3-(1-pyridiniummethyl)-cef-3-em-4-carboxylic acid dihydrochloride The product from (a) above (3,38 g) was dissolved in 98% formic acid (20 ml) 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 solution by evaporation under reduced pressure leaving an oil which was triturated with acetone (30ml) to give the title compound (2.20g). The τ(D ₂ O/NaHCO ₃ ) value is
3.08 (s, -thiazole proton), 1.06, 1.44,
1.93 (pyridinium proton), 4.16 (d, H5Hz,
7-H), 4.74 (d, J5Hz, 6-H), 8.55 (s, -
C(CH ₃ ) ₂ ). Acetone by nmr is 1 mole. Water content 5% (Karlfitscher method). Actual value of chlorine: 10.1% [C ₂₂ H ₂₄ N ₆ O ₇ S ₂ Cl ₂ +
Theoretical value of acetone (1 mol) + water (5%) Cl10.0
%] Example 10 a (6R,7R)-7-[(Z)-2-(2-triphenylmethylaminothiazol-4-yl)-
2-(2-tert-butoxycarbonylprop-
2-oximino)acetamide]-3-(1-
Pyridinium-methyl)-cef-3-em-4
-Carboxylate The product (2.18 g) from Preparation 10b was prepared in Example 9(a).
The title compound (4.03 g) was obtained, the spectroscopic properties of which were similar to those of the product of Example 9(a). b (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)acetamide]-
3-(1-Pyridiniummethyl)cef-3-em-4-carboxylic acid dihydrochloride The product from (a) above (3.8 g) was treated as in Example 9(b) to give the title compound (2.17 g) was obtained, but
Its spectroscopic properties were similar to those of the composition of Example 9(b). Example 11 a (6R,7R)-7-[(Z)-2-(2-triphenylmetalaminothiazol-4-yl)-
2-(1-tertiary-butoxycarbonylcyclobut-1-oximino)acetamide]-3-
(1-pyridiniummethyl)cef-3-m-
4-Carboxylate Phosphorus pentachloride (1.38g) was dissolved in 60ml dichloromethane. The solution was cooled to -10° and the product of Preparation 6 (3.48 g) was added in one portion. solution 30
Stir at -5° for minutes. Triethylamine (1.8ml)
Water (20ml) was then added. Mix at 0°C for 3 minutes
It was stirred with The lower phase was then treated with dimethylacetamide (30 ml) and acetonitrile (30 ml) with triethylamine (4.2 ml) added at -10°C.
The product of Preparation 10a (2.18 g) was added to the pre-chilled mixture in the medium. The reaction mixture was stirred at -5°C to 10°C for 45 minutes.
Cooling was then removed and the reaction 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.
The ethyl acetate extracts were combined, dried in the presence of charcoal and the solvent removed under reduced pressure. The residue was triturated with isopropyl ether to give the title compound (3.80g). νmax (nujiyor) 1780cm
^-1 (β-lactam), τ (CDCl ₃ ) 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-pyridinium Methyl) cef-3
-Em-4-carboxylic acid dihydrochloride The product from a above (2.75g) was stirred for 1 hour at ambient temperature in a mixture of 98% formic acid (15ml) and concentrated hydrochloric acid (0.9ml). The mixture was then filtered and the solvent removed under reduced pressure. The resulting residue was triturated with acene to produce the title compound (1.79g). νmax (Nujiol) 1785 cm ^-1 (β-lactam).
The τ (D ₂ O + NaHCO ₃ ) values are 1.05, 1.42, 1.91 (m,
pyridinium proton), 3.01 (s, aminothiazole proton), 4.13 (d, J5Hz, C ₇ proton),
4.68 (d, J5Hz, C-6 proton), 7.4-8.4 (broad m, cyclobutyl proton). Dimethylacetamide (1/3 mole) and acetone (1/2 mole) by nmr. Water content 7.4% (Karlfitscher method). Actual value of chlorine: 9.2% (C ₂₃ H ₂₄ N ₆ O ₇ S ₂ Cl ₂ +1/3
Molar dimethylacetamide + 1/2 molar acetone + theoretical Cl9.5% as 7.4% water) Example 12 (a) (6R,7R)-3-acetoxymethyl-7-
[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oximino)-acetamide]-cef-3-em-4
- Carboxylic hydrochloride The product of Example 1(a) (200 g)
It was dissolved in formic acid (800 ml) cooled to 10° and to this stirred mixture concentrated hydrochloric acid (60 ml) was added over 5 minutes. Stirring was continued at 20°-22° for 1 1/4 hours, then cooled to +10° and filtered.
The floor was washed with formic acid (30ml). The liquid and washings were combined and concentrated by evaporation at 20° to give a yellow foam which was triturated with ethyl acetate (800ml). The precipitated solid was collected by filtration, washed with ethyl acetate (200ml) and dried in vacuo at room temperature overnight to give the title compound (124.6g). λmax (ethanol) 234.5nm, E1% 1cm311. (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 The product from a above (40 g) was added to a stirred mixture of water (40 ml) and pyridine (25.6 ml), sodium iodide (160 g) was added and the mixture was dissolved in 3 1 / Heated at 60° for 2 hours. The hot solution was poured into stirred acetone () and diluted with diethyl ether (1.2). The suspension was cooled to 2° and the crude product was collected by filtration (50.65 g). This was dissolved in water (480 ml) and formic acid (19.3
ml), stirred with Amberlite LA2 (280ml) in ether (560ml). The mixture was separated and the organic layer was washed twice (240ml each) with water. The aqueous layer was washed with ether (280 ml) and applied to a column of zeolite 225SRC15' (200 ml H ⁺ ) and washed with distilled water until the eluate was 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 a syrup, which was added dropwise to stirred acetone (500ml). Filtration and equilibration in air gave the title compound (13.09g). H ₂ O 7.0% (Karl Fischer), λmax255nm (E1% 1cm364), λinf1 243
and 285 nm (E1% 1 cm338 and 171), [α] ²⁰ _D −3° (PH6 phosphate buffer). Example 13 (a) (6R,7R)-7-[(Z)-2-(2-tritylaminothiazol-4-yl)-2-(2-tert-butoxycarbonylprop-2-oximino)acetamide ]-3-(1-pyridiniummethyl)cef-3-em-4-carboxylate N,N-dimethylformamide solvate The micronized product of Example 9(a) was stirred at 23°N, N-dimethylformamide (15
ml). The solid dissolved and crystallization occurred after a while. The stirred mixture was diluted by portionwise addition of diisopropyl ether (20ml). The solid was collected by filtration to give the title compound (3.06 g) as colorless needle crystals. N,N-dimethylformamide by nmr = 2 1/2 mol. τ (DMSO-d ₆ ): 2.4-3.0 (m, trityl),
3.32 (s, aminothiazole ring proton), 0.47,
1.38, 1.82 (pyridinium proton), 4.34 (m, c
-7 proton), 4.92 (d, J-5, C-6 proton), 8.64 (s, tertiary butyl proton), 8.62
(s, (CH ₃ ) ₂ −C), [α] ²⁰ _D = −27.5° (c = 1
.1 in methanol). b (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(carboxyprop-2-oximino)-acetamide]-3-
(1-pyridiniummethyl)cef-3-m-
4-Carboxylic acid dihydrochloride The product from step a above (2.1 g) was dissolved in formic acid (10 ml).
It melted at 22°. Concentrated hydrochloric acid (0.8ml) was added and after 75 minutes the precipitated solid was removed. The liquid was evaporated and industrial denatured alcohol (10ml) 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 (1.35g). [α] ²⁰ _D
-14.7° (c=0.95, in PH6 buffer). τ(DMSO− _d6 ): 0.28(d, J9,

[Formula]), 0.77 (d, J6), 1.25 (t, J6), 1.70 (t, J6, pyridinium ring proton), 3.0 (s, aminothiazole proton), 3.99 (dd, J9.5, 7-H ), 4.67(d,
J5,6-H), 8.42(s,-( _CH3 ) ₂ ). Formulation examples are explained below. Example A (Dry powder for injection) Formulation per vial (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2-carboxyprop-2- Oximino)acetamide]-3
-(1-pyridinium methyl) cef-3-em-4-carboxylate 500 mg Lysine acetate 180 mg Cephalosporin antibiotic was blended with lysine acetate and filled into glass vials. The headspace of the vial was purged with nitrogen and the vial was sealed with a crimp. The product was dissolved for administration by adding 2 ml of water for injection. Example B (Dry powder for injection) (6R,7R)-7-[(Z) sterilized so that each vial contained an amount equal to 1.0 g of antibiotic acid.
-2-(aminothiazol-4-yl)-2-(2
-carboxyprop-2-oxyimino)acetamide]-3-(1-pyridiniummethyl)cef-
Fill 3-m-4-carboxylate monosodium salt 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) 500 mg sterile (6R, 7R)-7-[(Z)-2
-(2-aminothiazol-4-yl)-2-
(2-carboxyprop-2-oximino)
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, clarify by filtration and fill 2.15 ml into clean ampoules. Pass carbon dioxide through the contents of each ampoule for 1 minute before sealing. Each ampoule is sterilized in an autoclave and examined for clarity. (c) Reconstitute the cephalosporin antibiotic by dissolving it in 2.0 ml of the above sodium bicarbonate solution just before administration. Example D (Dry powder for injection) Formulation per vial (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclobut-1 -oximino)acetamide]
-3-(1-pyridiniummethyl)cef-3-
Em-4-carboxylate 500 mg Anhydrous sodium carbonate 47 mg Cephalosporin antibiotic was combined with sodium 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 adding 2 ml of water for injection. Example E (veterinary injection) Prescription (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclobuto-1-oximino)acetamide]
-3-(1-pyridiniummethyl)cef-3-
M-4-carboxylate 10% w/v Aluminum distearate 2% w/v Amount to make 100% w/v of both ethyl oleate...Amount to make the total 100% w/v Aluminum distearate to ethyl oleate Disperse in art and heat to 150° C. for 1 hour with stirring, then cool to room temperature. 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-pyridiniumethyl)cef-3
-Em-4-carboxylate. B: (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, sodium salt. C: (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclobuto-1-oximino)acetamide]-3-(1-pyridinium methyl)-cef-3-em-4-carboxylate. D: (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclobuto-2-oximino)acetamide]-3-(4-carbamoyl -1-pyridinium methyl)-cef-3-em-4-carboxylate, sodium salt. E: (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycycloprop-2-oximino)acetamide]-3-(1-pyridinium methyl)-cef-3-em-4-carboxylate, sodium salt. F: (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxycyclopent-1-yloxyimino)acetamide]-3-(1 -pyridinium methyl)
- cef-3-m-4-carboxylate,
Dihydrochloride. Test Method 1 Determination of Minimum Inhibitory Concentration (MIC)a MIC for all microorganisms except Haemophilus influenzae: Serial 2-fold dilutions of the freshly prepared test solution are prepared in casitone agar and poured into Petri dishes. added. Each plate was inoculated using a multipoint inoculator with an inoculum containing approximately 10 ⁵ colony forming units/ml of the microorganisms listed in the table. These microorganisms (but K.pneumoniae
1522E) are all clinical isolates.
μ after 18 hours incubation at 37°C
The MIC in g/ml was determined as the minimum concentration that inhibited growth. b MIC for Haemophilus influenzae
was measured by the broth dilution method in the same manner as in a above. However, brain heart infusion supplemented with fildes and yeast extract was used as the medium.

【table】

【table】

[Table] 2 Mouse protection test (ED ₅₀ ) 15% yeast suspension 0.5% to groups of 5 mice each
Bacterial challenge in ml was given intraperitoneally. Two treatment doses of test compound were administered subcutaneously as a solution in 0.2 ml of saline at 1 and 5 hours post-challenge.
Five days after challenge, the number of viable bacteria was counted and the activity of the test compound was expressed as the average effective dose. The E. coli strain used is a cephalosporin-sensitive strain. Table Mouse protection test (using E. coli 851) Test compound ED ₅₀ (mg/Kg/dose) A 1.0 B 0.6 C 0.5 D 0.5 E - F 1.5 3 β-lactamase stability (usually present in the range of 260-265 nm) (determined spectrophotometrically by following the ultraviolet absorption of the β-lactam ring and comparing its titer with cephaloridine (which is supposed to have a stability value of 1 for each enzyme). represent Results The test compound was stable against P99, K1 and TEM β-lactamase with a stability value of >30 for cephaloridine. 4 Toxicity a LD ₅₀ Values for Compounds A and C Aqueous solutions of the sodium salts of each compound were freshly prepared with sodium bicarbonate. 20 of each compound
% w/v solution to female CRH albino mice (body weight
16-22g) group and the _L50 value was measured. The results shown in the following table (a) were obtained. Table (a) Test compound LD ₅₀ (g/Kg) A 4.3 B 4.6 b Test compound was administered to mouse groups each consisting of 5 female CRH albino mice (body weight 18-22 g).
A single dose of 500 mg/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 above compound is 500mg/Kg or more.
Indicates that it has an LD ₅₀ value. Compounds C, D and E were not tested.

Claims (1)

[Claims] 1. General formula [In the formula, B is S or S→O, R ⁷ is an amino group or a protected amino group, and R ^8a
represents a hydrogen atom or a carboxyl-occluding functional group, or has a group of the formula -COOH and an associated anion A in place of the group -COO in the 4-position of the above formula. However, in the above formula, when B is S, R ^8a is a hydrogen atom, and R ⁷ is an amino group, the group at the 4-position in the above formula is COO ^- or
Not COOH. ] A cephalosporin compound or a salt thereof.