KR840000254B1 - Process for preparing heterocyclic derivatives of oxy-imino-substituted cephalosporins - Google Patents

Abstract

The cephalosporins[I; R=(CH2)nCO2R', NR'R", CONR'R" (R', R"=H, C1-6 alkyl); n=0, 1, 2; R1=H, amino-protective group; R2=H, OH-protective group, optionally substituted C1-6 aliphatic group; X=0, 1, 2 were prepd. Thus, 4.43g syn-III was converted into the acid chloride, which reacted with 3.8g II in Me2Co, followed by treatment with aq. HCO2H to give 1.13g syn-isomer 3-[(8-amino-6-tetrazolo-[1,5-b -pyridazinyl)-thiomethyl -7-[2-(2-amino-4-thiazolyl) ! -2-methoxyiminoacetamido -3-cephem-4-carboxylic acid.

Description

Method for preparing heterocycle derivative of ox-imino-substituted cephalosporin

The present invention relates to a process for the preparation of the novel, ox-imino-substituted cephalosporins of the following general formula (I).

그룹(여기에서 각각의 R' 및 R"는 같거나 다를수 있으며 수소원자 또는 C₁내지 C₆알킬그룹을 나타낸다)을 나타내며 ; R₁은 수소원자 또는 아미노-보호그룹을 나타내고 ; R₂는 수소원자, 하이드록시-보호그룹, 또는 하이드록시, 시아노, -COOR'(여기에서 R'는 상기 정의한 바와 같다) 및

(여기에서 R' 및 R"는 상기 정의한 바와 같다) 중에서 선택된 치환체에 의해 치환되거나 비치환될 수 있는, 직쇄 또는 측쇄의 포화 또는 불포화 C₁내지 C₆지방족 탄화수소그룹을 나타내며, x는 0,1 또는 2이다.Wherein R is a-(CH ₂ ) _n -COOR 'group where n is 0, 1 or 2, and R' represents a hydrogen atom or a C ₁ to C ₆ alkyl group, or

Group (where each R ′ and R ″ may be the same or different and represent a hydrogen atom or a C ₁ to C ₆ alkyl group); R ₁ represents a hydrogen atom or an amino-protective group; R ₂ represents a hydrogen atom , Hydroxy-protecting group, or hydroxy, cyano, -COOR ', wherein R' is as defined above; and

Wherein R ′ and R ″ are as defined above and represent a straight or branched, saturated or unsaturated C ₁ to C ₆ aliphatic hydrocarbon group which may be unsubstituted or substituted by a substituent selected from: x is 0,1 Or two.

The present invention also provides pharmaceutically and veterinary acceptable salts of compounds of general formula (I) as well as possible isomers such as syn- and anti-isomers, cis- and trans-isomers and optical isomers and mixtures thereof, antimicrobial activity It includes both metabolites having and metabolic precursors of general formula (I) compounds.

In the general formula of the present invention, the broken line (ξ) means that the oxyimino group has both syn- and anti-coordination.

As mentioned above, each of the neo- and anti-isomers of the compounds of formula (I) and mixtures thereof are all within the scope of the present invention.

The chain linked to the carbon atom at the 7-position is always the 7β-chain.

In the compound of formula (I), the 7β-chain can exist in either or both of the following tautomeric forms.

The present invention includes both compounds of the general formula (I) in which the 7β-chain is thiazolin form (IA) and compounds in which the 7β-chain is thiazole form (IB) as well as mixtures thereof.

When x is 1 the resulting compound is a sulfoxide and may be present in the R or S configuration. when x is 2 the compound is sulfone.

If R ₁ is an amino-protecting group it is for example formyl, optionally halogen-substituted C ₁ to C ₆ aliphatic acyl, preferably chloroacetyl or dichloroacetyl, tert-butoxycarbonyl, p-nitrobenzyl One of the protecting groups commonly used in peptide chemistry such as oxycarbonyl or trityl groups.

When R ₂ is a hydroxy-protecting group, it is for example formyl, acetyl, chloroacetyl, dichloroacetyl, trifluoroacetyl, tetrahydropyranyl, trityl or silyl group, especially trimethylsilyl or dimethyl-3 Tert-butylsilyl.

When R ₂ is a C ₁ to C ₆ aliphatic hydrocarbon group, it is preferably C ₁ to C ₆ alkyl, in particular C ₁ to C ₃ alkyl, or C ₂ or C ₃ alkenyl.

, -CH₂CN, -CH₂-CONH₂또는 -CH=CH-COOH이며 ; x는 0,1 또는 2인 일반식(Ⅰ) 화합물의 신-이성체 및 그의 약학적으로 또는 수의과적으로 허용되는 염이다.Preferred groups among the compounds of the present invention are those wherein R is -NH ₂ , -NHCH ₃ , -COOH, -CONH ₂ or -CH ₂ COOH; R ₁ is hydrogen; R ₂ is hydrogen, methyl, ethyl, —CH ₂ COOH or

, -CH ₂ CN, -CH ₂ -CONH ₂ or -CH = CH-COOH; x is a neo-isomer of the compound of formula (I) which is 0,1 or 2 and a pharmaceutically or veterinary acceptable salt thereof.

Particularly preferred compounds of the invention are those wherein R is -NH ₂ , -COOH or -CH ₂ COOH; R ₁ is hydrogen; R ₂ is hydrogen, methyl or ethyl; x is a neo-isomer of a compound of formula (I) which is 0 and a pharmaceutically or veterinary acceptable salt thereof.

More particularly preferred compounds of the invention are those wherein R is -NH ₂ or -COOH; R ₁ is hydrogen; R ₂ is methyl or ethyl; x is a neo-isomer of a compound of formula (I) which is 0 and a pharmaceutically or veterinary acceptable salt thereof.

Pharmaceutically and veterinary acceptable salts of the compounds of formula (I) include inorganic acids such as hydrochloric acid and sulfuric acid; Organic acids such as citric acid, tartaric acid, malic acid, maleic acid, mandelic acid, fumaric acid and methanesulfonic acid; Alkali metals, in particular sodium and potassium, alkaline earth metals, especially calcium and aluminum and alkali or alkaline earth metal carbonate salts (these alkali and alkaline earth metal salts can be formed by reaction with hydroxides, carbonates or bicarbonates, Salts are formed by reaction with aluminum hydroxide salts); Or organic amines, in particular lysine, triethylamine, procaine, dibenzylamine, N-benzyl-β-phenethylamine, (N, N'-dibenzyl) ethylenediamine, dehydroabiethylamine, N-ethylpy Salts with organic bases such as ferridine, diethanolamine, N-methylglucamine, tris (hydroxymethylamino) methane and the like. Intramolecular salts (ie, zwitterions) are also included within the scope of the present invention.

Specific examples of the compound of the present invention are as follows:

(1) 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydro Roxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(2) 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-meth Methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(3) 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-sia Nomethoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(4) 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-amino Carbonylmethoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(5) 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-carboxy Methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(6) 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydro Roxyiminoacetamido] -1 (R) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer);

(7) 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydro Roxyiminoacetamido] -1 (S) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer);

(8) 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-meth Methoxyiminoacetamido] -1 (R) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer);

(9) 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-meth Methoxyiminoacetamido] -1 (S) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer);

(10) 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydro Roxyiminoacetamido] -1-sulfonyl-3-cepem-4-carboxylic acid (new-isomer);

(11) 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-meth Methoxyiminoacetamido] -1-sulfonyl-3-cepem-4-carboxylic acid (new-isomer);

(12) 3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydro Roxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(13) 3-[(8-carboxy-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-meth Methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(14) 3-[(8-aminocarbonyl-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2 -Hydroxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(15) 3-[(8-aminocarbonyl-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2 -Methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(16) 3-[(8-methylamino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2- Hydroxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(17) 3-[(8-methylamino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2- Methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(18) 3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2- Methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(19) 3-[(8-carboxymethyl-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2- Methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer);

(20) 3-[(8-β-carboxyethyl-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl)- 2-methoxyimino-acetamido] -3-cepem-4-carboxylic acid (new-isomer); And pharmaceutically or veterinary acceptable salts thereof.

The structure of the compound of general formula (I) mentioned above is shown in the following table.

[table]

Compound of the present invention

A) reacting a compound of formula (II) or a reactive derivative thereof with a compound of formula (III) or a reactive derivative of compound of formula (III), removing the protecting group present if desired; or

B) reacting a compound of formula (IV) or a salt thereof with a compound of formula (V) or a reactive derivative of a compound of formula (V), removing the protecting group present if desired; or

C) Reacting a compound of formula (VI) or a salt or ester thereof with thiourea to give a compound of formula (I) wherein R ₁ is hydrogen, optionally removing / removing protecting groups present in R ₂ Or, if desired, convert the compound of formula (I) into a salt thereof or obtain a free compound from the salt, and / or if desired separate the isomeric mixture into a single isomer and / or optionally formula (I) a compound It can be prepared by a process characterized by the conversion to another compound of (I).

Wherein x, R, R ₁ and R ₂ are as defined above; E represents an amino group or a group of the formula -N = C = Y, wherein Y is oxygen or sulfur; Z represents a halogen atom.

In the general formulas (II), (IV) and (VI), the free carboxy group can be protected by conventional means before the start of the reaction, if necessary. Protective groups include groups commonly used in peptide synthesis such as, for example, tert-butyl, benzhydryl, p-methoxybenzyl, p-nitrobenzyl, trityl and trialkylsilyl. The protecting group can be removed after the reaction is completed by known methods, for example by mild acid hydrolysis or catalytic hydrogenation such as using Pd / C at atmospheric pressure. However, the removal of a protecting group is not an essential step of the process because the compound of formula (I) containing the protecting group is also included in the present invention.

In the starting materials used in each of the processes A) to C) mentioned above, if one or more asymmetric carbon atoms are present, they may be optically active compounds or racemic compounds. The starting material may also be syn- or anti-isomers and mixtures thereof, and may also be cis- or trans-isomers and mixtures thereof.

The reactive derivative of the general formula (II) compound may be, for example, an amine salt, silyl ester or metal salt. Reactive derivatives of the general formula (III) compounds are, for example, salts such as acyl halides, anhydrides, mixed anhydrides, azides, reactive esters or salts formed by alkali or alkaline earth metals, ammonia or organic bases. Reactive esters are, for example, p-nitrophenyl esters, 2,4-dinitrophenyl esters, pentachlorophenyl esters, N-hydroxysuccinimide esters and N-hydroxyphthalimide esters.

Z in the compound of formula (VI) is preferably chlorine or bromine.

Salts of compounds of formula (VI) are, for example, alkali or alkaline earth metal salts of the —COOH group in the compound. In addition, free carboxy, carboxymethyl or carboxyethyl groups, which may be represented by R substituents, may similarly form salts.

The reaction between the compound of formula (II) or a reactive derivative thereof and the compound of formula (III) or a reactive derivative thereof is acetone, dioxane, tetrahydrofuran, acetonitrile, chloroform, methylene chloride, N, N-dimethylformamide, In a suitable solvent such as 1,2-dichloroethane or a mixture of water and a solvent miscible with water, at room temperature or under cooling, preferably at about -50 ° C to about + 40 ° C, optionally sodium bicarbonate, It can be carried out in the presence of a base such as potassium bicarbonate or trialkylamine or in the presence of other acid acceptors such as alkylene oxides (eg propylene oxide).

When the compound of formula (III) is reacted with a compound of formula (II) wherein E in the free acid or salt form is amino, the reaction is preferably carried out in the presence of a condensing agent such as N, N'-dicyclohexylcarbodiimide. Do. After the reaction is completed, the protecting group can be optionally removed by known methods. For example, tert-butoxycarbonyl groups can be removed by treatment with an aqueous solution of an acid (eg HCl or H ₂ SO ₄ ), and monochloroacetyl groups can be removed with treatment with thiourea. Formyl and trifluoroacetyl groups can be removed by treatment with potassium bicarbonate in aqueous methanol, tetrahydropyranyl groups with dilute hydrochloric acid and trityl groups with formic acid or trifluoroacetic acid. have.

The reaction between the compound of formula (IV) and the heterocyclic thiol of formula (V) is carried out in the presence of about 2 equivalents of sodium bicarbonate in the presence of water or an organic solvent such as acetone, ethanol, dioxane or tetrahydrofuran. It can be carried out in a mixture with. The reaction temperature is preferably in the range of about 5 ° C. to about 90 ° C., and the pH is preferably maintained at about 5 to about 7.5. If desired, buffers such as sodium phosphate or acetate can be used. Alternatively, the same reaction can be carried out for a reaction time of several hours to several days at a temperature of about 50 ° C. to about 120 ° C., in base-free, completely anhydrous solvent. The preferred solvent is acetonitrile, and it may be ideal to carry out the reaction under inert atmosphere (eg nitrogen) to prevent oxidation of the heterocyclic thiols (V). Then, optionally, the protecting group can be removed by a known method as described above.

The reaction of the compound of formula (VI) or a salt or ester thereof with thiourea is preferably such as N, N-dimethylformamide, N, N-dimethylacetamide, dimethylsulfoxide, acetonitrile, hexamethylphosphorotriamide It is carried out in an aprotic solvent or a mixture of these solvents. The reaction temperature is preferably in the range of about 0 ° C to about 90 ° C. Subsequently, any removal of the protecting groups which may be present as R ₂ may be carried out as described above.

Random conversion of the compound of formula (I) to its salt and process for preparing the free compound from the salt or ester, as well as the process for the isolation of the monoisomers of the mixture of isomers and other compounds of the formula (I) of the compound of formula (I) Any conversion to the compound can be carried out by known methods. Thus, for example, a compound of general formula (I) where x is 1 and a sulfoxide is S-coordinated may contain the corresponding compound of general formula (I) where x is zero, as oxidizing agents, in particular perbenzoic acid, permaleic acid, sodium periodate, Treatment with peracids such as hydrogen peroxide or mixtures thereof, or with inorganic or organic acids such as formic acid, acetic acid or trifluoroacetic acid is preferably obtained. The reaction can be carried out in a solvent such as dioxane, tetrahydrofuran, chloroform, methylene chloride, formic acid, acetic acid, benzene, N, N-dimethylformamide, N, N-dimethylacetamide and the like. The reaction temperature is preferably about -30 ° C to about + 90 ° C.

In order to obtain compounds in which the sulfoxide is R-coordinated, for the intermediate product, preferably for compounds of general formula (II) wherein E is amino, the amino group is first protected by the formation of a Schiffbase and then the same oxidation Preference is given to carrying out the reaction. Schiffbases can be prepared by known methods, for example, by treating amines of general formula (II) with aldehydes such as benzaldehyde, salicylaldehyde or p-nitrobenzaldehyde, and after the oxidation reaction is completed, phenylhydrazine Treatment with hydrazine derivatives such as 2,4-dinitrophenylhydrazine, or Grignard reagents can generate free amino groups. The carboxyl group is preferably protected using the above-mentioned protecting group or the like during the oxidation reaction.

The process of converting sulfide to sulfoxide, i.e., compound of general formula (I), wherein x is 0, to the corresponding compound of x, can be carried out using 1 to 1.2 molar equivalents of oxidant per mole of compound to be oxidized.

The process of converting sulfides into sulfones, i.e., compounds of formula (I) with x of 0, and corresponding compounds of formula (I) with x of 2, oxidizing the same oxidizing agent as the oxidizing agent used to obtain the sulfoxide It can be carried out using at least 2 molar equivalents per mole.

Compounds of formula (II) wherein E is amino and x is 0 are prepared, for example, by reacting 7-amino-cephalosporanic acid or a salt thereof with a compound of formula (V) under reaction conditions well known in the literature. can do.

Compounds of the general formula (II) wherein E is -N = C = Y and x is 0 is a compound of the general formula (II) wherein E is amino and x is 0, using known methods, phosgene or thio It can be prepared by reacting with phosgene.

General formula (III) compound is a well-known compound or can be manufactured by a well-known method.

Compounds of general formula (IV) wherein x is 0 can be prepared, for example, using 7-amino-cephalospo, using reaction conditions similar to those described above in the reaction of compounds of general formula (II) It can be prepared by reacting lanic acid or a salt thereof with a compound of formula (III) or a reactive derivative thereof.

Compounds of general formula (VI) with x of 0 are general, for example, in which E is amino and x is 0 using reaction conditions similar to those described for the reaction of general formula (II) and general formula (III) compounds. The compound of formula (II) may be prepared by reacting with a compound of formula (VII) or a reactive derivative thereof such as one of the reactive derivatives mentioned above for the compound of formula (III).

Z is as defined above. The compound of formula (III) thus obtained is nitrosed using nitrosyl chloride or nitrosyl chloride, or an organic or inorganic nitrite such as amyl nitrite, sodium nitrite or potassium nitrite in the presence of an acid such as hydrochloric acid or acetic acid. _It can be converted into a compound of formula (VI) wherein _divalent hydrogen.

The nitrosification reaction is carried out at room temperature or cooling in a suitable solvent such as dioxane, acetonitrile, tetrahydrofuran, acetic acid or a mixture of one of these solvents with water, preferably at a temperature ranging from about -20 ° C to about 40 ° C. can do. The carboxyl groups present in the compound prior to the nitrosaturation may optionally be converted to salt form by treatment with alkali metal hydroxides or protected with the above-mentioned protecting group, after which the protecting group is removed by a known method after the reaction is completed. can do.

A compound represented by the general formula (Ⅵ) R ₂ is other than hydrogen may be obtained by conventional etherification or esterification to the corresponding compound of R ₂ is hydrogen in the starting material.

Compounds of formulas (II), (IV) and (VI) in which x is 1 or 2 can be obtained by oxidizing the corresponding compounds in which x is 0 as described above in the conversion to compounds of formula (I). have.

When the starting materials of formulas (III), (IV) and (VI) are neo-isomers, the reaction products are neo-isomers and vice versa. In most cases small amounts of anti-isomers are obtained with the neo-isomers. Isomers can be separated by known methods such as fractional crystallization or chromatographic methods.

Compounds of the invention are Staphylococcus, Streptoco-ccus, Diplococcus, Klebsiella, Escherichia coli, Proteus in animals or humans Only for Gram-positive and Gram-negative bacteria that are normally susceptible to cephalosporins, such as Proteus mirabilis, Salmonella, Shigella, Haemophilus and Neisseria But also high beta-lactamase producing gram-negative microorganisms such as Klebsiella aerogenes 1082E, Escherichia coli Tem, Enterobacter cloacae P99, Indole-positive Proteus, etc. Pseudomonas aeruginosa strains exhibit antimicrobial activity and are generally resistant to most cephalosporin compounds. Also has an antibiotic activity, therefore these compounds are gram-is particularly suitable for an infectious disease, for the treatment of urinary infections and respiratory infections caused by such bacteria.

In general, the activity of the compounds of the present invention on bacteria and beta-lactamase producing bacteria susceptible to cephalosporins is greater than that of cefazoline and cepuroxime. For example, compound 3-[(8-amino-6-tetrazolo [1,5-b] -pyrazazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl)- 2-methoxyacetamido] -3-cepem-4-carboxylic acid (new-isomer: named FCE 20127) is about 24 times more active than cefazoline against Streptococcus, and cepha also against most Gram-negative bacteria It exhibits about 30 times greater activity than sleepy and cefuroxime. In addition, compound FCE20127 shows about 20-60 times greater activity than cepuroxime against several beta-lactamase producing bacteria, such as Klebsiella aerogens 1082E, Enterobacter cloase P99, and Escherichia coli Tem. Compound FCE20127 was tested against 10 Pseudomonas aeruginosa strains, 4 Proteus vulgaris strains, and 4 Proteus morganii strains, confirming that they have several times greater activity than cefazoline and cepuroxime. .

Compound 3-[(8-carboxy-6-tetrazolo [1, 5-b] -pyridazinyl) thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyimine Noacetamido] -3-cefe-4-carboxylic acid (new-isomer: designated FCE 20485) is about 5 times more active against streptococcus than cefazoline, and for most Gram-negative bacteria About 32-fold, about 40-fold greater activity than cepuroxime. For bacteria such as Enterobater aerogenes ATCC 8308, Enterobacter Cloase 1321E, Salmonella typhi Watson, and Shigella sonnei ATCC 11060, compound FCE 20485 is more effective than cephazoline. There is 15 to 50 times greater activity than 50 to 100 times greater than cefuroxime.

Compound FCE 20485 also shows at least 100-500 fold greater activity against cephazoline and cefuroxime against Proteus vulgaris X20 and Proteus mirabilis ATCC 9921. Compound FCE 20485 also shows apparent in vitro activity against Pseudomonas aeruginosa G. and Bacteroides fragilis VPI 9032.

Another important characteristic of compound FCE 20485 is that the plasma half-life (mouse: about 90 minutes, rat: about 100 minutes) is very long in mice and rats after intravenous injection, whereas the plasma half-life of cefazoline in each case is different. At 16 minutes and 20 minutes. Thus, compound FCE 20485 can be used, for example, by E. coli G, Klebsiella pneumoniae ATCC 10031, Proteus mirabilis ATCC 9921, E. coli Tem., Haemophilus influenzae. ), In vivo tests with mice infected with Salmonella typhi watson and Proteus vulgaris X20, show 20-200 times greater activity than cefazoline.

There is little toxicity of the compounds of the present invention and therefore can be used safely in treatment. For example, the approximate acute toxicity (LD ₅₀ ) of compounds FCE 20127 and FCE 20485 measured 7 days after treatment with intravenous administration of increased doses of mice is greater than 2000 mg / kg. Similar activity and toxicity data are measured for other compounds of the invention.

Therefore, the compounds of the present invention may include respiratory infections (eg bronchitis, bronchitis, pleurisy), hepatobiliary system and abdominal infections (eg cholecystitis, peritonitis), blood and cardiovascular infections (eg sepsis), urinary tract infections (eg pyelonephritis). , Cystitis), gynecological infections (e.g. cervicitis, endometritis), otolaryngitis (e.g. otitis, sinusitis, mumps), are useful for the treatment of infections caused by Gram-positive or Gram-negative bacteria . The compounds of the present invention can be administered to humans or animals in various dosage forms, for example orally in the form of tablets, capsules, loadings or syrups; Administered rectally in the form of suppositories; Parenteral administration (by intravenous administration in an emergency), by intravenous or intramuscular (in solution or suspension) administration; Inhalation in the form of an aerosol or solution using a nebulizer; Vaginally administered in the form of a bougie; Or topically in the form of lotions, creams and ointments.

The present invention includes pharmaceutical or veterinary compositions containing a compound of the present invention and a pharmaceutically or veterinary acceptable excipient. Such compositions may be prepared by conventional methods using conventional excipients, generally carriers and / or diluents, used in other cephalosporin compositions. Typical carriers or diluents are, for example, water, gelatin, lactose, starch, magnesium stearate, talc, vegetable oil, cellulose and the like. The daily dose is about 1 to about 100 mg / kg body weight and the exact dose for various types of animals depends on the age, weight and condition of the subject being treated and the frequency and route of administration. The preferred mode of administration of the compounds of the present invention is parenteral administration, in which case the compounds are sterile water or lidocaine hydrochloride solution and intravenous sterile water, physiological saline solution, dextrose solution or conventional intravenous liquid or electrolyte. It is dissolved in the adult patient can be administered 1 to 4 times a day in an amount of about 100mg to 200mg, preferably about 150mg once.

In addition, the compound of the present invention is a compound which is suspended or dissolved in a conventional inert anhydrous or aqueous carrier for washing or spraying at a concentration of about 0.2 to 1% by weight of the compound, as an antimicrobial agent used for prophylactic purposes such as cleaning or surface sterilization composition. Can be used as These compounds are also useful as nutritional supplements in animal feed.

The following examples illustrate the invention. In some cases, the compound tends to contain a solvent, making the melting point somewhat difficult.

) 중에서 측정하며, UV스펙트럼은 pH7.4완충인산염 용액 중에서 측정하거나 바우쉬-롬브(Bausch-Lomb) 장치상 1% NaHCO₃중에서 측정한다. NMR스펙트럼은 내부표준으로 (CH₃)₄Si를 사용하여 DMSO(디메틸설폭사이드) 또는 CDCl₃중에서 최종생성물에 대해서는 브루커(Bruker) HX-90(90MHz)으로, 중간체 생성물에 대해서는 퍼킨-엘머(Perkim-Emer) R-24B(60MHz)로 측정한다.The IR spectrum is in the solid phase (KBr) or on the Perkin-Elmer 125 spectrometer.

UV spectra are measured in pH 7.4 buffered phosphate solution or 1% NaHCO on a Bausch-Lomb device.₃Measure in. NMR spectrum is internal standard (CH₃)₄DMSO (dimethylsulfoxide) or CDCl using Si₃The final product is measured with Bruker HX-90 (90 MHz) for the final product and Perkim-Emer R-24B (60 MHz) for the intermediate product.

Example 1

3-[(8-amino-6-tetrazolo- [1,5-b] -pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxy Iminoacetamido] -3-cepem-4-carboxylic acid (new-isomer)

Anhydrous methylene chloride containing 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid (new-isomer; 4.43 g; 0.01 mol) and triethylamine (1.41 ml; 0.01 mol) 70 ml) of the stirred solution was cooled to 0 ° C and phosphorus pentachloride (2.08 g; 0.01 mol) was added little by little. After stirring at 0 ° C. for 10 minutes and at room temperature for 1 hour, the mixture is evaporated under reduced pressure and re-evaporated in acetone / benzene to remove residual traces of phosphorus oxychloride and dissolved in 50 ml of anhydrous acetone. It is then filtered to remove triethylamine hydrochloride. The acetone solution of 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetylchloride thus obtained was purified by 7-amino-3-[(8 in a mixture of water and acetone (75ml: 50ml). -Amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -3-cepem-4-carboxylic acid (3.8 g; 0.01 mol), NaHCO ₃ (0.84 g) and triethylamine (2.82 ml) was added dropwise to the vigorously stirred ice cold solution. The resulting slurry was stirred at 0 ° to 5 ° C. for 0.5 hours, then stirred at room temperature for 1.5 hours, dissolved in ethyl acetate (500 ml), washed with dilute hydrochloric acid (50 ml), washed with aqueous sodium chloride (100 ml) and sodium sulfate Dried to evaporated to dryness. The residue was stirred in methylene chloride (5 ml), then ethyl acetate (50 ml) and diethyl ether (50 ml) were added. The precipitated product is collected, dissolved in warm methylene chloride (30 ml), treated with diethyl ether (100 ml) with cooling and stirring. After stirring for 20 minutes, the white solid was filtered, washed with ether and dried to give 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7 as a white powder. -[2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) is obtained.

The above-mentioned compound (2.0 g) is added in portions to a stirred heat (55 ° C.) solution of formic acid (13 ml) in water (13 ml). Stirring and heating are continued for 25 minutes at the same temperature, then the solid is filtered from the cooled mixture. The solid is triturated in 99% formic acid (10 ml), then water (2 ml) is added and pure triphenylmethanol is filtered off.

The acid mother liquors are combined and evaporated to give a residue. After complete removal of water and formic acid, polishing with anhydrous ethanol yields a solid (1.225 g). The crude product is suspended in water (70 ml) and a sufficient amount of NaHCO ₃ is added to produce a solution. The pH of the solution was lowered to 2.0 by adding 2N HCl, and after 20 minutes, the precipitated product was collected, washed thoroughly with water, and dried at 75 ° C. for 18 hours. This gives 1.13 g of the title compound which decomposes without melting at above 225 ° C.

Elemental analysis: C ₁₈ H ₁₇ N ₁₁ O ₅ S ₃

Found: C 37.90; H3.14; N26.78; S16.83

Calculated: C 38.35; H3.04; N27.33; S17.07

IR (KBr) [cm ^-1 ]: 3320-3180NH

1760> C═O (β lactam), 1520-CONH- (secondary amide), 1030 = NO-CH ₃

NMR, 100 MHz (DMSO-d ₆ ): [δ, ppm] 3.74 (2H, dd, 2-CH ₂ ) J = 17 Hz, 3.88 (3H, S., -OCH ₃ ), 4.36 (2H, dd, 3- CH ₂ S) J = 13 Hz, 5.20 (1H, d., 6 -H) J = 5 Hz, 5.83 (1H, dd, 7-H) J = 5 Hz + 8 Hz, 6.41 (1H, S., pyridazine) 7-H), 6.82 (1H, S., 5-H in a thiazole ring), 6.80 to 7.80 (2H, broad S., -NH _{2 in a} thiazole ring), 8.02 (2H, broad S., pyridazine Cyclic -NH ₂ ), 9.69 (1H, d., -CONH-) J = 8Hz

Example 2

2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid and 7-amino-3-[(8-aminocarbonyl-6-tetrazolo using the method described in Example 1 [1,5-b] -pyridazinyl) thiomethyl] -3-cepem-4-carboxylic acid was reacted to produce 3-[(8-aminocarbonyl-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl- [7- [2- (2-amino-thiazolyl) -2-methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) To obtain.

Elemental analysis: C ₁₉ H ₁₇ N ₁₁ O ₆ S ₃

Found: C38.71; H2.96; N25.81; S16.01

Calculated: C38.57; H2.89; N26.04; S16.26

TLC (CHCl ₃ : MeOH: HCOOH: H ₂ O = 140: 75: 20: 20): Rf = 0.42

I.R. (KBr) <C = O β-lactam

The 7-amino-3-[(8-aminocarbonyl-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -3-cepem-4-carboxylic acid used as starting material was Manufacture. : 6-chloro-8-aminocarbonyl-tetrazolo [1,5-b] pyridazine

3-hydrazino-4-aminocarbonyl-6-chloro-pyridazine (4.2 g; 0.0244 moles) in 15% acetic acid. See J. Het. To an ice cold solution prepared by the method described in Chemistry, 7, 465 (1970), a solution of sodium nitrite (1.55 g) in water (10 ml) is added dropwise. After stirring at 5-10 ° C. for 1 hour, the separated precipitate is filtered off, crystallized with water, filtered again and dried at 50 ° C. in vacuo to yield 3.44 g (77.5%) of the title compound at a melting point of 226 ° C. (decomposition).

Elemental Analysis: C ₅ H ₃ ClN ₆ O

Found: C30.11; H1.44; N42.00; Cl17.69

Calculated value: C30.24; H1.52; N42.32; Cl17.85

6-mercapto-8-aminocarbonyl-tetrazolo [1,5-b] pyridazine

To a stirred solution of NaSH.H ₂ O (10 g; 0.135 mol) in 150 ml of water was added 6-chloro-8-aminocarbonyl-tetrazolo [1,5-b] pyridazine (10 g; 0.05 mol) and the mixture was added. Stir vigorously for 90 minutes. After cooling to 0 ° C., the reaction mixture is acidified with 23% HCl. After cooling for 1 hour at 0 ° C., the solids are collected and washed with a small amount of cold water to give the title compound in quantitative yield with a melting point of 185 to 187 ° C. (decomposition).

IR (New sol): -SH 2480cm ^-1 ,> C = O 1675cm ^-1

TLC (CHCl ₃ : CH ₃ OH: HCOOH = 160: 70: 30): Pure product

UV (phosphate buffer at pH 7.4): λ _max = 262;

Elemental Analysis: C ₅ H ₄ N ₆ OS

Found: C30.65; H2.00; N42.53; S16.16

Calculated value: C30.60; H2.05; N42.84; S16.34

7-amino-3-[(8-aminocarbonyl-6-tetrazolo [1,5-b] -pyridazinyl) -thiomethyl] -3-cepem-4-carboxylic acid

6-mercapto-8-aminocarbonyl-tetrazolo- [1,5-b] pyridazine (2 g; 0.010 mol) and 2.8 g sodium bicarbonate in 90 ml of phosphate buffer (pH6.4) (50-55 ° C) To the solution is added 7-ACA (4.6 g; 0.017 mol) in portions and the mixture is heated at 60 ° C. for 4 hours. After cooling at 10 ° C., precipitated 7-amino-3-[(8-aminocarbonyl-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -3-cepem-4-carboxylic acid Is collected by filtration. The solid is suspended in 150 ml of Me ₂ CO: H ₂ O (1: 2) and stirred for 30 minutes to refilter. The precipitates are combined, washed with Me ₂ CO and dried under vacuum at 60 ° C. to give the title compound (80%) with a melting point of 228 to 230 ° C. (decomposition).

Elemental Analysis: C ₁₃ H ₁₂ N ₈ O ₄ S ₂

Found C37.93; H2.93; N27.13; S15.45

Calculated value C38.22; H2.96; N27.43; S15.70

IR (New sol):> C = O β-lactam 1780cm ^-1

UV (1% NaHCO ₃ ): λ _max = 249; λ _max = 332

Example 3

Using the same method, the following compounds are obtained.

3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Cetamido] -1 (R) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer); 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Cetamido] -1 (S) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer); 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Cetamido] -1-sulfinyl-3-cepem-4-carboxylic acid (new-isomer); 3-[(8-methylamino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyimine Noacetamido] -3-cepem-4-carboxylic acid (new-isomer).

Example 4

3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer).

Anhydrous methylene chloride containing 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid (new-isomer; 11.35 g; 0.0256 moles) and triethylamine (3.61 ml; 0.0256 moles) 200 ml) of the stirring solution was cooled to 0 ° C and phosphorus pentachloride (5.6 g; 0.027 mol) was added little by little. After stirring at 0 ° C. for 15 minutes and at room temperature for 1 hour, the mixture is evaporated under reduced pressure, dissolved in anhydrous benzene and redevaporated to remove traces of phosphorus oxychloride. This treatment is repeated twice (2 x 50 ml). The residue was suspended in 50 ml of anhydrous acetone, and triethylamine hydrochloride was removed by filtration. The acetone solution of 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetylchloride thus obtained was diluted with 7-amino-3-[(8 in a mixture of water and acetone (500 ml: 250 ml). Carboxy-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -3-cepem-4-carboxylic acid (10 g; 0.0232 mol) and NaHCO ₃ (15 g) dropwise to a vigorously stirred ice cold solution do. The mixture is stirred for 30 minutes at 0-5 ° C., then for 90 minutes at room temperature, the undissolved material is filtered off and evaporated under vacuum to remove acetone. The aqueous phase is adjusted to pH 2 by addition of 8% HCl, extracted with ethyl acetate (3 × 400 ml), washed with aqueous sodium chloride, dried (Na ₂ SO ₄ ) and evaporated to dryness. The residue was stirred with diethyl ether, stirred for 20 minutes, the white solid was filtered off, washed with ether and dried to give 3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -Thiomethyl] -7- [2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cepem-4-carboxylic acid is obtained. The resulting compound is added portionwise to a stirred heat (55 ° C.) solution of 50% formic acid (140 ml). After stirring and heating at the same temperature for 30 minutes, the solid is filtered from the cooled mixture. The filtrate is evaporated to dryness in vacuo to give a residue. After complete removal of water and formic acid, grinding with ether gives a solid. Crystallization of the crude product with 50% ethanol yields 9.1 g (60%) of the title compound which decomposes at 225 ° C.

Elemental analysis: C ₁₉ H ₁₆ N ₁₀ S ₃ O ₇

Found C38.10; H2.91; N23.34; S15.93

Calculated value C38.50; H2.72; N23.63; S16.23

TLC (CHCl ₃ : MeOH: HCOOH: H ₂ O = 140: 75: 20: 20): Rf = 0.25

IR (KBr) ν 3500-2300 cm ^-1 Combined -COOH, ν1765 cm ^-1 > C = O β-lactam, ν1710 cm ^-1 C = O acid, 1650 cm ^-1 secondary, 1620-1580 cm ^-1 {(C = N) oxime, ν (CN) + δ (NH)}, 1540 cm ^-1 ν (CN) + δ (NH) amide NMR, 100 MHz (DMSO-d ₆ ): δ p.pm 3.68 (1H, d, 2-CH) ₂ ); 3.86 (1H, d, 2-CH ₂ ); 3.92 (3H, S, = N-OCH ₃ ); 4.36 (1H, d. 3-CH ₂ -S); 4.69 (1H, d, 3-CH ₂ -S); 5.21 (1H, d, 6-H); 5.85 (1H, doublet of doublets, 7-H); 6.83 (1H, S, 5-H in a thiazole ring); 7.30 (2H, br-s, -NH _{2 on} thiazole ring); 8.02 (1H, S, 7-H in pyridazine ring); 9.83 (1 H, d, -CONH).

7-Amino-3-[(8-carboxy-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -3-cepem-4-carboxylic acid used as starting material was prepared as follows. : 3-hydrazino-4-carboxy-6-chloro-pyridazine

A mixture of 3,6-dichloro-4-carboxy-pyridazine (20 g; 0.103 mol) and 22 ml of 98% hydrazine hydrate in 200 ml of 50% ethanol was refluxed for 1 hour with stirring. After cooling to 5 ° C., the solid precipitates were collected and washed with 20 ml of anhydrous ethanol. The solid was suspended in 100 ml of water and the mixture was adjusted to pH 1-2 with 23% HCl, cooled to 5 ° C., then the solid was filtered and dried in vacuo at 80 ° C. to yield 18.29 g (93.4 g) of the title compound at a melting point of 198 to 201 ° C. %) Is obtained.

Elemental Analysis: C ₅ H ₅ ClN ₄ O ₂

Found C31.64; H2.64; N29.32; Cl18.53

Calculated value C31.84; H2.67; N29.71: Cl18.80

NMR (DMSO-d ₆ ): 7.8 (1H, s, 5-H in pyridazine ring), 9.2 (4H, br-s, -COOH, -NHNH ₂ )

6-chloro-8-carboxy-tetrazolo [1,5-b] pyridazine

To a ice cold suspension of 3-hydrazino-4-carboxy-6-chloro-pyridazine (1.88 g; 0.01 mol) in 15% acetic acid is added dropwise a solution of sodium nitrite (0.70 g) in water (5 ml). After stirring for 1 hour at 5 to 10 ° C., the separated precipitate is filtered off and washed with anhydrous ethanol. The solid is suspended in 2N HCl and the mixture is stirred for 30 minutes, then the solid is filtered and washed with cold water and dried in vacuo at 70 ° C. to give 1.73 g (87%) of the title compound at a melting point of 225 ° C.

Elemental Analysis: C ₅ H ₂ ClN ₅ O ₂

Found C29.95; H0.98; N35.15; Cl17.58

Calculated value C30.09; H1.00; N35.09; Cl17.76

IR (New sol): <C = O 1730cm ^-1

NMR (DMSO-d ₆ ): 8.33 (1H, S, 7-H in pyridazine ring)

6-mercapto-8-carboxy-tetrazolo [1,5-b] pyridazine

6-Chloro-8-carboxy-tetrazolo [1,5-b] pyridazine (10 g; 0.05 mol) was added to a stirred solution of NaSH.H ₂ O (10 g; 0.135 mol) in 150 ml of water, and the mixture was stirred for 2 hours. Stir vigorously. After cooling to 0 ° C., the reaction mixture is acidified with 23% HCl. After cooling at 0 ° C. for 1 hour, the resulting solids are collected and washed with a small amount of cold water to give 9 g (91.5%) of the title compound having a melting point of 210 ° C. (decomposition).

Elemental Analysis: C ₅ H ₃ N ₅ O ₂ S

Found: C30.41; H1.52; N35.61; S16.19

Calculated value: C30.45; H1.53; N35.52; S16.26

IR (KBr): (SH) 2540cm ^-1 ,> C = O 1725cm ^-1

UV (phosphate buffer solution at pH 7.4): λ _max = 258,

7-Amino-3-[(8-carboxy-6-tetrazolo [1,5-b] -pyridazinyl) -thiomethyl] -3-cepem-4-carboxylic acid

6-mercapto-8-carboxy-tetrazole [1,5-b] pyridazine (8.2 g; 0.0415 mol) and 11.6 g sodium bicarbonate in 265 ml of phosphate buffer (pH6.4) 7-ACA (18.5 g; 0.068 mol) is added in portions and the mixture is heated at 60 ° C. for 5 hours with stirring. After cooling to 20 ° C., the undissolved material is filtered and the solution is adjusted to pH 4.4 by addition of 23% HCl. The precipitate is filtered off and washed with a mixture of Me ₂ CO: H ₂ O (3: 1) and then with Me ₂ CO to give the sodium salt (75%) of the title compound at melting point> 270 ° C. (decomposition).

Elemental analysis: C ₁₃ H ₁₀ N ₇ O ₅ S ₂ Na

Found: C35.80; H2.66; N22.43; S14.66; Na5.10

Calculated: C36.19; H2.33; H22.72; S14.86; Na5.32

IR (New sol):> C = O β-lactam 1770cm ^-1

UV (1% NaHCO ₃ ): λ _max = 246;

The sodium salt is suspended in water and acidified with 8% HCl, the resulting solid is filtered and washed with water to give the title compound having a melting point of 245 ° C. (decomposition).

Elemental analysis: C ₁₃ H ₁₁ N ₇ O ₅ S ₂

Found: C38.45; H2.90; N23.55; S15.10

Calculated: C38.13; H2.70; N23.94; S15.66

Example 5

3-[(8-amino-6-tetrazolo [1, 5-b] -pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyimine Noacetamido] -3-cepem-4-carboxylic acid (new-isomer)

N, N in a cold (0 ° to 5 ° C.) solution of 2- (2-tritylamino-4-thiazolyl) -2-methoxyimino-acetic acid (3.675 g; 0.0828 moles) in anhydrous methylene chloride (13 ml) '-Dicyclohexylcarbodiimide (0.9 g; 0.00438 mol) was added and stirred at 40C for 40 minutes and at room temperature for 30 minutes, and the resulting slurry was diluted with anhydrous methylene chloride. The separated solid, ie dicyclohexylurea (0.98 g; quantitative yield), is filtered off and washed with fresh methylene chloride. The methylene chloride solution is combined, evaporated to dryness and the resulting symmetric anhydride of the starting acid is immediately dissolved in anhydrous acetone (30 cc). The obtained solution was taken as 7-amino-3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -3-cepem- in 1: 1 aqueous acetone (120 ml). To the cooled solution in an ice bath of 4-carboxylic acid (1.65 g; 0.00436 mol), triethylamine (1.22 ml; 0.00872 mol) and NaHCO ₃ (0.347 g; 0.00414 mol) is added dropwise with stirring. After stirring for 1 hour at 0 ° to 5 ° C., the ice bath is removed and after a further 2 hours the mixture is dissolved in ethyl acetate (500 ml) and shaken with aqueous 1N HCl (100 ml). The insoluble material (a small amount of unreacted 7-amino-cefem) was filtered off and the organic layer was washed with aqueous sodium chloride solution, dried over Na ₂ SO ₄ , filtered and evaporated. Most of the starting 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid contained in the crude reaction mixture was precipitated from dioxane solution (30 ml) by the slow addition of dicyclohexylamine (1.4 ml). Let's do it. After 20 minutes at 12 ° C., the resulting dicyclohexylamine salt was filtered off, the solution was diluted with ethyl acetate (200 ml), acidified with 1N HCl, and the aqueous layer was discarded. The organic phase was washed with aqueous sodium chloride solution, dried over Na ₂ SO _4, concentrated to reduce the volume and added to diethyl ether to 3-[(8-amino-6-tetrazolo [1,5-b] -pyridazinyl) 3.1 g of -thiomethyl] -7- [2- (2-tritylamino-4-thiazolyl) -2-methoxy-iminoacetamido] -3-cef-4-carboxylic acid (new-isomer) To obtain. The product was treated with 50% aqueous formic acid as described in Example 1 to afford the title compound showing the same TLC, IR and NMR data as the product prepared in Example 1.

Using the same method, the following compounds are prepared: 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino- 4-thiazolyl) -2-cyano-methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-aminocarbonylmethoxy Iminoacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-carboxymethoxyimine Noacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxy-imi Noacetamido] -1 (R) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer); 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Cetamido] -1 (S) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer); 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Setamido] -1-sulfonyl-3-cepem-4-carboxylic acid (new-isomer); 3- [8-aminocarbonyl-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl-2-methoxyimine Noacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-methylamino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-Amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer).

Example 6

3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer)

N, N 'in a cold (0 ° to 5 ° C) solution of 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid (3.675 g; 0.0828 moles) in anhydrous methylene chloride (13 ml) Dicyclohexylcarbodiimide (0.9 g; 0.00438 mol) was added and stirred at 40C for 40 minutes and at room temperature for 30 minutes, and the resulting slurry was diluted with anhydrous methylene chloride. The separated solid, ie dicyclohexylurea (0.98 g; quantitative yield), is filtered off and washed with fresh methylene chloride. The methylene chloride solution is combined, evaporated to dryness, and the resulting symmetric anhydride of the starting acid is immediately dissolved in anhydrous acetone (30 cc). The obtained solution was taken as 7-amino-3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -3-cefem in 1: 1 aqueous acetone (120 ml). To the cooled solution in an ice bath of -4-carboxylic acid (1.78 g; 0.00436 mol), triethylamine (1.22 ml; 0.00872 mol) and NaHCO ₃ (0.347 g; 0.00414 mol) was added dropwise with stirring. After stirring for 1 hour at 0 ° to 5 ° C., the ice bath is removed and after a further 2 hours the mixture is dissolved in ethyl acetate (500 ml) and shaken with aqueous 1N HCl (100 ml). The insoluble material (a small amount of unreacted 7-amino-cefem) was filtered off and the organic layer was washed with aqueous sodium chloride solution, dried over Na ₂ SO ₄ , filtered and evaporated. Most of the starting 2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetic acid contained in the crude reaction mixture was precipitated from dioxane solution (30 ml) by the slow addition of dicyclohexylamine (1.4 ml). Let's do it. After 20 minutes at 12 ° C., the resulting dicyclohexylamine salt is filtered and the solution is diluted with ethyl acetate (200 ml) and acidified with 1N HCl and the aqueous layer is discarded. The organic phase was washed with aqueous sodium chloride solution, dried over NaSO _4, concentrated to reduce the volume and added to diethyl ether to 3-[(8-carboxy-6-tetrazolo [1,5-b│pyridazinyl) -thiomethyl 3.3 g of] -7- [2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) are obtained. The product was treated with 50% aqueous formic acid as described in Example 4 to give the title compound showing the same TLC, IR and NMR data as the product prepared in Example 4.

Example 7

3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydroxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer)

2- (2-tritylamino-4-thiazolyl) -2-trityloxyiminoacetic acid (2.25 g; 0.00335 moles) and triethylamine (0.47 ml; 0.00335 moles) in anhydrous methylene chloride cooled to -5 ° C. To the solution of was added phosphorus pentachloride (0.697 g; 0.00335 mol) at once. After stirring at −5 ° C. for 20 minutes and at room temperature for 1 hour, the reaction mixture is evaporated under reduced pressure without external heating until all POCl ₃ is removed. The residue was dissolved in anhydrous acetone (50 ml) and triethylamine hydrochloride was filtered off. The acetone solution of 2- (2-tritylamino-4-thiazolyl) -2-trityloxyiminoacetylchloride thus obtained was cooled to 0 ° C. and 7-amino- in water (35 ml) and acetone (25 ml). 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -3-cef-4-carboxylic acid (0.76 g; 0.002 mol), triethylamine (0.562 ml; Add dropwise to a solution containing 0.004 moles and NaHCO ₃ (0.281 g; 0.00335 moles).

After addition, the slurry is stirred at 0 ° C. for 30 minutes and at 25 ° C. for 90 minutes. The reaction mixture was poured into ethyl acetate (350 ml) with vigorous stirring, water (50 ml) was added, and a sufficient amount of 2N HCl was added to adjust the pH of the aqueous phase to two baths. The organic phase is separated, washed with aqueous sodium chloride solution, dried over Na ₂ SO ₄ and evaporated to dryness. The resulting foamed material is triturated with diethyl ether to afford 1.98 g of crude product containing a significant amount of starting acid. Most of these impurities are removed by dissolving the crude in dioxane (10 ml) and adding the resulting solution dropwise to diethyl ether (70 ml). After stirring for 10 minutes, the white precipitate was collected by filtration to collect 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-trityl Amino-4-thiazolyl) -2-triyloxy-iminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) (1.02 g) is obtained.

The prepared compound (1 g) was added to 50% aqueous formic acid solution (40 ml) maintained at 55 ° C. (oil bath) with stirring. After 35 minutes, the mixture is cooled to 25 ° C. and suction filtered to wash the solid with fresh 50% formic acid (20 ml), then with distilled water and discard. The acidic solutions are combined and evaporated under reduced pressure. The residue was dissolved in 99% ethanol and evaporated again. In 99% ethanol, it was evaporated to a small volume (5 ml) and filtered. The resulting beige powder is dissolved in 2% aqueous NaHCO ₃ (20 ml) and charcoal is added. The filtered solution is adjusted to pH 2 by adding 2N HCl and stirred for 5 minutes. The precipitate was collected by filtration, washed thoroughly with water, washed with a small amount of ethanol, dried at 65 ° C. for 16 hours, and degraded without melting at about 205 ° C. in 3-[(8-amino-6-tetrazolo [ 1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydroxyiminoacetamido] -3-cepem-4-carboxylic acid ( Neo-isomer) 0.25 g.

Elemental analysis: C ₁₇ H ₁₅ N ₁₁ O ₅ S ₃

Found: C36.81; H2.88; N27.73; S16.92

Calculated: C37.15; H2.75; N28.03; S17.20

IR (KBr) [cm ^-1 ]: 3400 -NH, 3000 -OH, 1760> C = O (β lactam)

NMR, 100 MHz (DMSO-d ₆ )

17Hz ; 4.16(1H, d., 3-CH₂S) ; 4.60(1H, d, 3-CH₂S) J

13Hz ; 5.21(1H, d., 6-H) ; 5.86(1H, d,d., 7-H) ; 6.42(1H, s., 피리다진환상의 7-H) ; 6.76(1H, s., 티아졸환상의 5-H) ; 7.20(2H, 브로드 s., 티아졸환상의 -NH₂) ; 8.02(2H, 브로드 s., 피리다진환상의 -NH₂) ; 9.56(1H, d., -CONH-) ; 11.66(1H, 브로드 s.,=N-OH)[δ, ppm]: 3.61 (1H, d., 2-CH ₂ ); 3.89 (1H, d., 2-CH ₂ ) J

17 Hz; 4.16 (1H, d., 3-CH ₂ S); 4.60 (1H, d, 3-CH ₂ S) J

13 Hz; 5.21 (1H, d. 6-H); 5.86 (1H, d, d., 7-H); 6.42 (1H, s., 7-H on pyridazine); 6.76 (1H, s., 5-H in a thiazole ring); 7.20 (2H, broad s., -NH _{2 on} thiazole ring); 8.02 (2H, broad s., -NH _{2 in} pyridazine ring); 9.56 (1 H, d., -CONH-); 11.66 (1H, broad s., = N-OH)

Using the same method, the compounds (6), (7), (10), (12), (14) and (16) of the above-described table are prepared.

Example 8

3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer)

3-acetoxymethyl-7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) in water (10 ml) (0.456 g; 0.001 mol) and NaHCO ₃ (0.143 g; 0.0017 mol) were added 8-amino-6-mercapto-tetrazolo [1,5-b] pyridazine (0.118 g; 0.0007 mol). do. Slow nitrogen flows through the solution and the mixture is heated at 50 ° C. for 12 hours while maintaining pH6.8. After cooling, the solution is acidified with 10% HCl and the precipitates are collected by filtration, washed with water and then with ethanol. This gave 0.215 g of compound (2), which exhibited the same NMR, TLC and IR data as the product described in Example 1, and a small amount of anti-isomer as impurities. Using the same method, the following compounds are prepared: 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino- 4-thiazolyl) -2-methoxyiminoacetamido] -1 (R) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer); 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Cetamido] -1 (S) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer); 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Setamido] -1-sulfonyl-3-cepem-4-carboxylic acid (new-isomer); 3-[(8-aminocarbonyl-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxy Iminoacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-methylamino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyimine Noacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-ethoxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-carboxymethyl-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyimine Noacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-β-carboxyethyl-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-meth Toxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer).

Example 9

3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer)

3-acetoxy-methyl-7- [2- (2-tritylamino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cepem-4-carboxylic acid in anhydrous acetonitrile (50 ml) 8-amino-6-mercapto-tetrazolo [1,5-b] pyridazine (0.336 g; 0.002 mol) was added to a stirred solution of (0.697 g; 0.001 mol) and the resulting slurry was stirred for 24 hours under a slow nitrogen stream. Reflux for a while. After adding an additional amount of mercapto compound (0.168 g; 0.001 mol) and stirring for additional 16 hours at reflux temperature, the mixture was cooled to filter out the undissolved substance (unreacted mercapto compound) and evaporated the acetonitrile solution. The resulting foamed material is taken up in warm methylene chloride (5 ml) and diethyl ether (25 ml) is added. The resulting white precipitate was collected by filtration, washed with a small amount of ethyl acetate and washed with diethyl ether to obtain 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl]. 0.650 g of -7- [2- (2-tritylamino-4-thiazolyl) -2-methoxyimino-acetamido] -3-cepem-4-carboxylic acid (new-isomer) is obtained. The product was treated with 50% aqueous formic acid as described in Example 1 to yield the title product, which exhibits the same TLC, IR and NMR data as the product prepared in Example 1.

Using the same method, the following compounds are prepared: 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4 -Thiazolyl) -2-hydroxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydroxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-aminocarbonyl-6-tetrazolo [1, 5-b] -pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydro Roxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-aminocarbonyl-6-tetrazolo [1, 5-b] -pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-meth Methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-methylamino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydroxyimine Noacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-methylamino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyimine Noacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-ethoxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-carboxymethyl-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyimine Noacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-β-carboxyethyl-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-meth Toxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer).

Example 10

3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer)

3-acetoxymethyl 7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) in water (10 ml) ( 8-carboxy-6-mercapto-tetrazolo [1,5-b] pyridazine (0.138 g; 0.0007 mol) was added to a stirred solution of 0.456 g; 0.001 mol) and NaHCO ₃ (0.143 g; 0.0017 mol). . Slow nitrogen flows through the solution and the mixture is heated at 50 ° C. for 12 hours while maintaining pH6.8. After cooling, the solution is acidified with 10% HCl and the precipitates are collected by filtration, washed with water and then with ethane. This gave 0.245 g of the title compound and a small amount of anti-isomers as impurities which showed the same NMR, TLC and IR data as the product described in Example 4.

Example 11

3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer)

3-acetoxymethyl-7- [2- (2-tritylamino-4-thiazolyl) -2-methoxy-iminoacetamido] -3-cepem-4-carboxylic acid in anhydrous acetonitrile (50 ml) 8-carboxy-6-mercapto-tetrazolo [1,5-b] pyridazine (0.394 g; 0.002 mol) was added to a stirred solution of (0.697 g; 0.001 mol) and the resulting slurry was stirred for 24 hours under a slow nitrogen stream. Reflux for a while. After adding an additional amount of mercapto compound (0.168 g; 0.001 mol) and stirring for an additional 16 hours at reflux temperature, the mixture was cooled to filter off the undissolved substance (unreacted mercapto compound) and evaporated the acetonitrile solution. The resulting foamed material was taken up in warm methylene chloride (5 ml) and diethyl ether (25 ml) was added. The resulting white precipitate was collected by filtration, washed with a small amount of ethyl acetate and washed with diethyl ether to give 3-[(8-carboxy-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] 0.730 g of -7- [2- (2-tritylamino-4-thiazolyl) -2-methoxyimino-acetamido] -3-cepem-4-carboxylic acid (new-isomer) is obtained. The product was treated with 50% aqueous formic acid as described in Example 4 to give the title product, which showed the same TLC, IR and NMR data as the product prepared in Example 4.

Example 12

3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydroxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer)

Step A: 4-Chloroacetoacetic acid

After ethyl 4-chloroacetoacetate (30 g; 0.182 mol) was stirred with 37% aqueous HCl (200 ml) for 15 hours at room temperature, the reaction mixture was poured into ice / water, extracted with ethyl acetate and dried over Na ₂ SO ₄ . Evaporation to dryness without external heating yielded 17.4 g (70% yield) of 4-chloroacetoacetic acid. The crude product is used in the next step without further purification.

NMR 60 MHz (CDCl ₃ ): 3.71 (2H, s., COCH ₂ COOH); 4.20 (2H, s., ClCH ₂ CO); 9.98 (1H, broad s., COOH)

Step B: 4-Chloroacetoacetylchloride

Oxalyl chloride (1.71 ml; 0.02 mol) was added to a solution of 4-chloroacetoacetic acid (2.4 g; 0.017 mol) in anhydrous benzene, followed by addition of 1 drop of N, N-dimethylformamide. After stirring at 25 ° C. for 15 hours, the solution is evaporated to dryness, dissolved in anhydrous benzene and evaporated again to yield 2.7 g of a black oily product for use in the next acylation step (step C). Alternatively, 4-chloroacetoacetylchloride is described in Helv. Chim. Acta. 60, 1256 (1977), can be prepared by reacting diketene with chlorine in a suitable inert solvent such as methylene chloride.

Step C: 3-[(8-amino-6-tetrazolo [1,5-b] -pyridazinyl) -thiomethyl] -7- [4-chloro-3-oxobutyramido) -3- Sepem-4-carboxylic acid

7-amino-3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -3-cef-4-carboxylic acid (3.8 g; 0.01 mol) was dried with anhydrous N, Suspend in N-dimethylformamide (50 ml) and add N, O-bis- (trimethylsilyl) -acetamide (8.13 g; 0.04 mol). After stirring for 2 hours at room temperature almost all starting material is dissolved. To the cooled (-30 ° C.) mixture is added dropwise a solution of 4-chloroacetoacetylchloride (2.35 g; about 0.015 moles) in anhydrous methylene chloride (20CC). The mixture is stirred at −30 ° C. for 2 hours and further raised to room temperature over 1 hour, after which isopropanol (40 ml) is added to decompose the silylated compound. The solvent is evaporated under reduced pressure and the oily residue is partitioned between water and ethyl acetate. The organic layer was separated, washed with aqueous sodium chloride solution, concentrated under reduced pressure to reduce the volume, and diethyl ether was added to complete the formation of the precipitate. The precipitated solid is collected by filtration, washed with fresh ether and dried in vacuo to give 4.6 g of the title compound which is used in the next step without further purification.

NMR 60 MHz (DMSO-d ₆ )

[δ, ppm]: 3.6-3,7 (4H, s + d, d., 2-CH ₂ + COCH ₂ CO); 4.35 (2H, doublet of 3-CH ₂ S); 4.57 (2H, s., Cl-CH ₂ CO); 5.08 (1H, d. 6-H); 5.69 (1H, doublet of doublets, 7-H); 6.3 (1H, S., 7-H in pyridazine ring); 7.95 (2H, broad NH. -NH _{2 on} pyridazine ring); 9.02 (1H, s., CONH)

Step D: 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- (4-chloro-2-hydroxyimino-3-oxobutyramimi Fig. 3)-Sepem-4-carboxylic acid (new-isomer)

3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- (4-chloro-3-oxobutyramido) -3 in acetic acid (60 ml) To a stirred solution of cefem-4-carboxylic acid (6.72 g; 0.0171 mol) is added a solution of sodium nitrite (1.65 g; 0.0239 mol) in water (6 ml) at 5 ° to 10 ° C. After stirring at room temperature for 3.5 hours, the reaction mixture is treated with brine and extracted with ethyl acetate (4 x 200 ml). The combined extracts are washed with water, dried over Na ₂ SO ₄ and evaporated to give a foam. The product was triturated with diethyl ether to afford 6.3 g of the title compound as a beige powder.

TLC (silica gel plate, CHCl ₃ / MeOH / HCOOH 160: 40: 20) identifies a single spot with an Rf value slightly smaller than the starting material.

NMR ,, 60 MHz (DMSO-d ₆ )

[δ, ppm]: 3.60 (2H, doublet of 2-CH ₂ ); 4.35 (2H, doublet of 3-CH ₂ S); 4.74 (2H, s., ClCH ₂ CO); 5.08 (1H, d. 6-H); 5.70 (1H, doublet of doublets, 7-H); 6.38 (1H, s., 7-H on pyridazine); 7.96 (2H broad s, -NH _{2 in} pyridazine ring); 9.21 (1H, d., -CONH-); 13.00 (1H, s., = N-OH)

Step E: 3-[(8-amino-6-tetrazolo- [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2- Hydroxy-iminoacetamido] -3-cepem-4-carboxylic acid (new-isomer), compound (1)

3-[(8-amino-6-tetrazolo [1, 5-b] -pyridazinyl) -thiomethyl] -7- (4-chloro-2-hydroxy-imino-3-oxobutyramimi Fig. 3)-Sepem-4-carboxylic acid (5.12 g; 0.01 mol) is dissolved in anhydrous N, N-dimethylacetamide (25 ml). Thiourea (0.76 g; 0.01 mol) is added and the mixture is stirred at room temperature for 3 hours. The resulting solution was added dropwise to ethyl acetate (250 ml) with stirring. The rubbery material precipitates, the supernatant is discarded and the residue is carefully ground with fresh ethyl acetate until a powder is formed. The powder is collected by filtration and dried. This affords N, N-dimethylacetamide solvate of the hydrochloride of the desired product. The resulting material is dissolved in aqueous NaHCO ₃ and washed five times with ethyl acetate and then with diethyl ether. After nitrogen was introduced for 15 minutes and charcoal was added, the charcoal was filtered off and the solution acidified with 2N HCl. The crystalline precipitates were collected, washed thoroughly with water and then washed with ethyl alcohol to obtain 3.8 g of the title compound which appeared to be the same as the product obtained in Example 7 by TLC, IR and NMR.

Using the same method, the following compound is prepared: 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4 -Thiazolyl) -2-hydroxyiminoacetamino] -1 (R) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer); 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydroxyiminoa Cetamido] -1 (S) -sulfinyl-3-cepem-4-carboxylic acid (new-isomer); 3-[(8-amino-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydroxyiminoa Cetamido] -1-sulfinyl-3-cepem-4-carboxylic acid (new-isomer); 3-[(8-carboxy-6-tetrazolo [1, 5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydroxyiminoa Cetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-aminocarbonyl-6-tetrazolo [1, 5-b] -pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydro Roxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer); 3-[(8-methylamino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-hydroxyimine Noacetamido] -3-cepem-4-carboxylic acid (new-isomer).

Example 13

3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2 in water (80 ml) A stoichiometric amount of NaHCO ₃ is added to an aqueous suspension of -methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) (5.63 g) to give a complete solution of the compound. The solution was lyophilized to yield 3-[(8-amino-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2 having a melting point of> 240 ° C (decomposition). A sodium salt of -amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) is obtained.

Elemental Analysis: Found Na3.80

Calculation value Na3.90

IR (KBr): 1760 cm ^-1 > C = O (β-lactam)

Example 14

3-[(8-carboxy-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2 in water (80 ml) 2 equivalents of NaHCO ₃ is added to an aqueous suspension of -methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) (5.92 g) to obtain a complete solution of the compound. The resulting solution was then lyophilized to afford 3-[(8-carboxy-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (with melting point> 270 ° C. (decomposition). A bisodium salt of 2-amino-4-thiazolyl) -2-methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) is obtained.

Elemental Analysis: Found Na6.91

Calculation Na7.19

IR (KBr): 1765 cm ^-1 > C = O (β-lactam)

Example 13

3-[(8-carboxy-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2 in acetone (400 ml) 2 equivalents of a 30% solution of sodium 2-ethylhexanoate in isopropyl alcohol are added to a solution of -methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) (5.92 g). After stirring for 30 minutes at room temperature, the mixture was diluted with petroleum ether and the resulting precipitate was filtered to give 3-[(8-carboxy-6-tetrazolo [1,5-b] pyridine having a melting point of> 270 ° C (decomposition). Disodium) -thiomethyl] -7- [2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido) -3-cefe-4-carboxylic acid (new-isomer) To obtain a salt.

Elemental Analysis: Found Na6.91

Calculation Na7.19

IR (KBr): 1765 cm ^-1 > C = O (β-lactam)

Example 16

Injectable pharmaceutical compositions include 3-[(8-carboxy-6-tetrazolo [1,5-b] pyridazinyl) -thiomethyl] -7- [2- (2-amino-4-thiazolyl)- 100-500 mg of 2-methoxyiminoacetamido] -3-cepem-4-carboxylic acid (new-isomer) disodium salt is prepared by dissolving in sterile water or sterile physiological saline (1-2 ml).

Claims (1)

Reacting a compound of formula (II) or a reactive derivative thereof with a compound of formula (III) or a reactive derivative thereof, or a compound of formula (IV) or a salt thereof Or a compound of formula (VI), or a salt or ester thereof, with thiourea to give a compound of formula (I) wherein R ₁ is hydrogen, or the compounds of formula (I) thus produced A process for preparing a compound of formula (I) and its optical isomers and pharmaceutically and veterinary acceptable salts, characterized by removing the protecting groups present at

Wherein R is a-(CH ₂ ) _n -COOR 'group, where n is 0, 1 or 2, and R' represents a hydrogen atom or a C ₁ to C ₆ alkyl group. or

Group, wherein each R ′ and R ″ may be the same or different and represent a hydrogen atom or a C ₁ to C ₆ alkyl group; R ₁ represents a hydrogen atom or an amino-protecting group; R ₂ is Hydrogen atom, hydroxy-protecting group, or hydroxy, cyano, -COOR ', wherein R' is as defined above; and

Wherein R ′ and R ″ are as defined above; a straight or branched, saturated or unsaturated C ₁ to C ₆ aliphatic hydrocarbon group which may be unsubstituted or substituted by a substituent selected from: x is 0,1 Or 2, E represents an amino group or a group of the general formula —NC = Y, wherein Y represents oxygen or sulfur, and Z represents a halogen atom.