NO794177L - CEPHALOSPORIN ANALOGS AND PROCEDURES FOR THEIR PREPARATION - Google Patents

Description

Cephalosporin analog compounds.

The present invention relates to hitherto unknown cephalosporin analogue compounds, more specifically hitherto unknown carba-cephem compounds which differ from the cephalosporins in that they have a carbon atom instead of the sulfur atom in the dihydrothiazine ring. In "Journal of the American Chemical Society", 96, 7584 (1974) and "J. Med. Chem.", 20, 551 (1977) certain carbacephems with substituted methyl groups in the 3-position, such as ( ±)-1-carba-cephalotin with the formula:

indicated to have antibacterial activity. In addition, some compounds that are included in the definition of the carbazefem compounds herein, which are described below, are mentioned as starting materials in US patent no. 4,123,528. In this patent, however, no physical data is given for these compounds.

As microorganism populations develop immunity to existing antibiotics, new antibacterial compounds are needed. For this purpose, hitherto unknown carbazephems with a hydrogen atom in the 3-position have been synthesized which are useful as intermediates in the preparation of compounds with unexpectedly high antibacterial activity, and such compounds and methods for their synthesis are stated in DE-OS no. 2,911,786 and No. 2,911,787.

According to the invention, it has now been shown that it is possible to synthesize hitherto unknown carbazephems with a halogen atom in the 3-position, which are also useful as intermediates in the preparation of compounds with unexpectedly high antibacterial activity.

In general, the invention relates to hitherto unknown cephalosporin analogues with the general formula:

where X means amino, azido or protected amino, Hal means halogen, and R means carboxyl or protected carboxyl.

(In the following, compounds with the above formula are sometimes referred to simply as compound I, and this also applies with regard to other formulas.)

More specifically, the invention relates to hitherto unknown cephalosporin analogues with the general formula I-1:

where Hal and R have the above meaning, and also

•compounds with the general formula 1-2:

where X"'" means azido or protected amino and Hal and R have the meaning given above, as well as salts thereof.

As protective groups in the "protected amino" specified for X or X"*" can e.g. mention is made of phthaloyl, t-butyloxycarbonyl, 2,2,2-trichloroethyloxycarbonyl, 2,2,2-trifluoroethyloxycarbonyl, benzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, phenylacetyl, phenoxyacetyl, benzylidene and salicylidene. ;Chlorine, bromine and iodine are suitable as halogens. The "protected carboxyl" indicated for R is represented by -COOR, where R is one. commonly used protecting group for a carboxyl group in the synthesis of penicillins and cephalosporins. As R 2 can e.g. alkyl with 1-5 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and t-butyl, halogenated alkyl with 1-5 carbon atoms are used. such as chloromethyl, 2,2,2-trichloroethy 1 and 2,2,2-trifluoroethyl, arylmethyl with 7-20 carbon atoms, such as benzyl, diphenylmethyl and triphenylmethyl, arylmethyl with 8-20 carbon atoms and optionally substituted with methoxy, nitro, etc. .on the phenyl ring, such as p-nitrophenylmethyl and p-methoxyphenylmethyl, substituted silyl such as trimethylsilyl and triphenylsilyl, and a group readily removed enzymatically or nonenzymatically in vivo, such as -CHOCOR 4, ;' 3 ;R ;where R 3 means hydrogen or alkyl with 1-6 carbon atoms,;4 ;and R means alkyl with 1-6 carbon atoms, alkoxy with 1-6 carbon atoms or phenyl. As salts of compound I can be mentioned e.g. salts of inorganic acids such as hydrochloride, sulphate, phosphate and carbonate, salts of organic acids such as formate, acetate, trifluoroacetate, malate and tartrate, and when R is carboxyl, salts with inorganic bases such as sodium, potassium, calcium and barium salts, and salts with organic bases such as salts with triethylamine. Compound I includes all stereoisomers in the 6- and 7-positions. A compound obtained by converting X in the general formula I for a cis-isomer to the enacyl-amino group exhibits stronger antibacterial activities than that obtained in the same way from the corresponding trans-isomer. Therefore, the cis isomers that can be converted into the most effective antibiotics are most applicable. Nevertheless, trans isomers are also useful since they can be converted into useful antibiotics by introducing an alpha-methoxy group at the 7-position. ;Compound I-1, corresponding to formula I, where X is amino, can be prepared from the compounds 1-2-2 stated below by the following reaction process I. ;Reaction process I ; ; where X is azido or phthaloylamino, and R and Hal have the above meaning. ;The reduction of the compound 1-2-2, where X^aer. azido, is carried out in a conventional manner in the synthetic chemistry of penicillins or cephalosporins. In this reaction, compound I-1 can be prepared by choosing suitable conditions and reagents to avoid degradation of substituents or functional groups in the carbacefem molecule. ;Suitable reduction methods include: 1) Catalytic reduction, 2) reduction using hydrogen sulfide and a base, 3) reduction using sodium borohydride, 4) reduction using zinc acid, and 5) reduction using chromium (II)- chloride as listed below. ;1. Catalytic reduction; Compound .1-2-2 is subjected to catalytic, reduction; in a stream of hydrogen gas in the presence of a catalyst in an inactive solvent to form compound I-1. Any solvent that does not affect the reaction can be used. Ethanol, water, tetrahydrofuran, dioxane, diethyl acetate, acetic acid or a mixture thereof are preferably used. Palladium-charcoal, platinum oxide, palladium-calcium carbonate and Raney nickel can be suitably used as catalyst. The reaction is usually carried out at a temperature of from -20 to 100°C, preferably at room temperature and a pressure of 1-50 atm, preferably atmospheric pressure. If this reaction uses a compound with the general formula 1-2-2, where P is carboxyl protected with a protective group such as substituted arylmethyl, • e.g. benzyl, paramethoxybenzyl, paranitrobenzyl, benzidryl and trityl, as starting material, a compound of the general formula I-1, where R is carboxy, can also be obtained. ;2. Reduction using hydrogen sulfide base; Compound 1-2-2 is reduced with hydrogen sulfide in the presence of a base in an inactive solvent to form compound I-1. As a solvent, e.g. methylene chloride, chloroform, benzene or tetrahydrofuran alone or in combination. As a base, triethylamine, pyridine and the like can be suitably used. The reaction is carried out at a temperature of 0-50°C, preferably at room temperature. ; 3. Reduction using sodium borohydride ; Compound 1-2-2 is reduced with sodium borohydride in an inactive solvent to form compound I-l. As a solvent, e.g. methanol, ethanol, dioxane or tetrahydrofuran alone or in combination. Sodium borohydride is used in an amount of 1 equivalent or in excess in relation to compound 1-2-2. The reaction is carried out at a temperature of 0-100°C, preferably 10-50°C. ; 4. Reduction using zinc acid; Compound 1-2-2 is reduced with zinc acid in an inactive solvent to form compound I-1. As a solvent, e.g. acetone, water, dioxane, tetrahydrofuran, ethanol or acetic acid, alone or in combination. As an acid, hydrochloric acid or acetic acid is suitable. The zinc and the acid are used in an amount of 1 equivalent or in excess in relation to compound 1-2-2. ;The reactions are carried out at a temperature of 0-100°C, usually from room temperature to 60°C. ;5. Reduction using chromium (II) chloride; Compound 1-2-2 is reduced with chromium (II) chloride in the presence of an acid in an inactive solvent. The acid, the solvents and the reaction conditions are the same as in the zinc-acid method. ;The removal of a phthaloyl group from compound 1-2-2 is carried out in a conventional manner to. removal of a phthaloyl group. As means of removal, e.g. hydrazine, hydroxylamine, ethylamine, dimethylamino-propylamine or sodium sulphide-methylhydrazine are used. The reaction can be carried out at from -20 to 30°C in an alcohol, such as methanol and ethanol, but other conventional conditions can also be used. ;Compound 1-2 is produced from compounds with the general formula ; ; in which x\ Hal and R have the above meaning, Y means a sulfur or selenium atom and R means an optionally substituted alkyl, aralkyl or aryl group or a heterocyclic group, according to the reaction process below; II. ;Reaction process II; ; This reaction consists in the removal of R<1->Y-OH and is usually carried out in a solvent. The reaction temperature is 0-2.00°C. As solvents, preferably inert solvents are used such as aliphatic hydrocarbons (hexane, heptane, etc.), halogenated hydrocarbons (chloroform, methylene chloride, carbon tetrachloride, etc.), aromatic hydrocarbons (benzene, toluene, xylene, etc.), ethers (diethyl ether, tetrahydrofuran, dimethoxyethane, etc. .), esters (ethyl acetate, etc.), amides (dimethylformamide, dimethylacetamide, etc.) and sulfoxides (dimethyl sulfoxide, etc.). ;In the case of the sulfoxide (Y=S), the reaction takes place by ■heating. The reaction temperature is preferably 50-200°C. If it is a matter of removing a phenylsulfinyl group, the best results are obtained by heating to the reflux temperature. In the case of the selenium oxide (Y=Se), the reaction takes place at a lower temperature than for the sulphoxide and the temperature is preferably 0-30°C. ;Under the definition of R"'' in the 'general formula - II, the alkyl group can be elucidated by an alkyl group with 1-5 carbon atoms such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl; the aralkyl group can is represented by phenylalkyl with 7-20 carbon atoms such as benzyl and phenethyl; and the aryl group can be represented by phenyl. Substituents in the alkyl, aralkyl or aryl group can be mentioned, for example, halogen such as chlorine or bromine, nitro, alkyl with 1- 4 carbon atoms and alkoxy with 1-4 carbon atoms. Furthermore, the heterocyclic group can be elucidated by tetrazolyl or thiazolyl. Preferably phenyl, Y means sulfur and Hal chlorine. Compound II includes all the stereoisomers at 2-, 3-, 6- and the '7-positions and mixtures thereof. Compound II in which the 6- and 7-positions are in the cis configuration is the most useful as a starting compound for the most useful compound .I in the cis configuration. Nevertheless, compounds II with trans -configuration useful as output connection for connect else I with trans configuration. ;Compound II is also a hitherto unknown compound and is prepared from a compound with the general formula ;III: ; ; in which , Y, R"1" and - R have the meaning stated above, according to reaction process III below. ;Reaction process III; ; This reaction consists in the introduction of a halogen atom on the alpha carbon atom of the sulfinyl or seleninyl group. E.g. the reaction to alpha-chlorination of the sulfoxide is carried out using a halogenating agent such as tosyl chloride, nitrosyl chloride, dichloroiodophenyl, t-butyl hypochlorite, sulfuryl chloride, preferably sulfuryl chloride, if necessary in the presence of a base, such as pyridine. The reaction is carried out at from -78-20°C, preferably under cooling in an inert solvent without water. The reaction time is usually within 30 minutes. As an example of a preferred embodiment, the reaction takes place very gently and rapidly using sulfuryl chloride as halogenating agent and a halogenated hydrocarbon such as dichloromethane as solvent to obtain the desired compound in good yield. "Addition of calcium oxide to the reaction system" to capture hydrogen chloride formed during the course of the reaction has a good effect in some cases. When an excess of sulfuryl chloride is used, dichloro compounds are sometimes formed as by-products. The reaction of alkyl phenyl sulfoxide or dialkyl sulfoxide with sulfur furyl chloride has been previously reported (K. C. Tin et al., "Tetrahedrbn-letter", 4643, 1970), but there is no report of such a reaction being applied to an alpha-lactan compound. Compound III is also a hitherto unknown compound and is produced from a compound with the general formula: ; wherein x"1", Y, R and R<1> have the above meaning, according to the following reaction process IV: ;Reaction process IV; ; This reaction consists in an oxidation of sulfide or selenide and conventional oxidation conditions for such compounds can be used to form a sulfinyl or seleninyl compound. Different oxidizing agents can be used such as manganese dioxide, chromic acid, lead tetraacetate, ruthenium tetroxide, N-halogen carboxamide, oxygen and ozone. Anodic oxidation, periodic acid, hydrogen peroxide and organic peracids such as m-chloroperbenzoic acid can be mentioned as particularly preferred oxidizing agents. ;Op the solvent is chosen according to the oxidase j onsrhidlet. Water, alcohol, acetic acid and mixtures thereof, or halogenated hydrocarbons such as chloroform, are preferred. . The reaction is usually carried out at from -50-100°C. When an excess of the oxidizing agent is used and the temperature is raised, sulfone is sometimes formed. ;Compound IV is also a previously unknown compound and is produced from a compound with the general formula: ; where X"*" and R have the meaning stated above, according to reaction process V below.

Reaction process V

This reaction consists in the addition of a thiol or selenol with the formula R^-Y-H. The reaction is usually carried out at from -50-100°C in a solvent and if necessary in the presence of a base.

As the base can e.g. organic bases such as piperidine, diethylamine, triethylamine are used; metal hydrides such as sodium hydride and potassium hydride; alkyllithium such as n-butyllithium; or sodium hydroxide, potassium hydroxide, etc. When a strong base is used, it is preferred, in order to avoid cleavage of the 3-lactan ring, that the compound R^-Y-H is first treated with 1 equivalent or less of the base and then reacted with compound V. It better results are obtained by using a weak base than by using a strong base. For example, better results are obtained by using piperidine when thiophenol is used as R"^-Y-H.

Solvents are used as solvents which are inactive towards the addition reaction. It can, e.g. aromatic hydrocarbons such as benzene, toluene, oxylene are used; halogenated hydrocarbons - such as chloroform, methylene chloride and carbon tetrachloride; ethers such as diethyl ether, tetrahydrofuran and dimethoxyethane; esters such as ethyl acetate; alcohols such as methanol and ethanol; amides such as dimethylformamide and dimethylacetamide; sulfoxides such as dimethyl sulfoxide; and acetonitrile.

Compound I in which R is carboxyl can be prepared by the following reaction processes VI using a compound of the general formula 1-3 as starting compound.

Reaction process VI

where X, Hal and R 2 have the meaning indicated above.

The reaction can be carried out in a conventional manner within the synthesis of penicillins or cephalosporins. In the reaction, compounds 1-4 can be prepared by choosing suitable conditions and reagents to avoid degradation of substituents or functional groups in the carbacefem molecule.

Suitable reactions that convert -COOR 2 to -COOH include: 1) Catalytic reaction, 2) acidolysis, 3) cleavage with a Lewis acid,-. 4) hydrolysis, 5) reduction other than catalytic reduction using reducing agents and 6) use of an esterase. Each of these reactions is described in more detail below.

1) Catalytic reduction

2

In this reaction, the group COOR is converted to the group COOH in the presence of a catalyst in a hydrogen atmosphere and an inactive solvent. Any solvent which does not affect the reaction and preferably ethanol, water, tetrahydrofuran, dioxane, ethyl acetate or acetic acid can be used as solvent. alone or in combination. As a catalyst, palladium-carbon, platinum oxide, palladium-calcium carbonate and Raney nickel are suitable. The reaction is usually carried out at a pressure of 1-50 times atmospheric pressure and a temperature of 0-100°C, preferably at atmospheric pressure and temperature.

This method is preferably used when R 2 is benzyl, p-nitrobenzyl, diphenylmethyl, triphenylmethyl, p-methoxybenzy1 or the like.

When X is an azido group, the azido group can be reduced to an amino group, when R 2 is converted to H by catalytic reduction. The resulting compound with an amino group is also a desired compound according to the invention.

2) Azidolysis

In this reaction, the group COOR 2 is converted to COOH with acid in an inactive solvent. As acids, hydrogen chloride, p-toluenesulfonic acid and trifluoroacetic acid are suitable. Any solvent which does not affect the reaction can be used as a solvent, preferably ethyl acetate, benzene, ethanol, acetic acid, dioxane, methylene chloride and chloroform, alone or in combination.

The reaction is usually carried out at a temperature of from -15-50°C, preferably 0-25°C, for from 10 min. to 5 hours, preferably from 30 min. to 3 hours. This method

is preferably used when R 2 is t-butyl, trityl, etc.

3) Cleavage with a Lewis acid

In this reaction, the group COOR 2 is converted to the group COOH by cleavage in the presence of a Levi's acid in an inactive solvent which can be any that does not affect the reaction, preferably a mixture of a nitroalkane such as nitromethane and a haloalkane such as methylene chloride. As a Lewis acid, e.g. aluminum chloride, boron trifluoride, titanium tetrachloride or tin tetrachloride. The acid is used in an amount of 1.0-1.5 molar equivalents in relation to compound 1-3. The reaction is preferably carried out in the presence of an agent such as anisole, which takes up any carbonium cation that may be formed. The reaction is carried out at a temperature of 0-50°C, preferably at room temperature for 1-10 hours.

Addition of a thiol compound such as ethanethiol, •

ethanedithiol, etc., often enhance this type of reaction.

This method is used -preferably when R 2 is benzyl, p-nitrobenzy1, etc.

4) Hydrolysis

2

In this reaction, the group COOR is converted to the group COOH by hydrolysis in the presence of an acid or base in an inactive solvent. Suitable acids include e.g. p-toluenesulfonic acid, hydrochloric acid and acetic acid. Any solvent which does not affect the reaction can be used and preferably methanol, N,N-dimethylformamide or acetic acid/water/tetrahydrofuran are usually used. The reaction is usually carried out at a temperature of 0-50°C, preferably 15-25°C for from 10 min. to 2 hours. This method with the use of an acid is preferably used when R 2 is t-butyldimethylIsilly.

Calcium carbonate is preferably used as alkali in an amount of 1-6 molar equivalents in relation to compound 1-3, and any solvent which does not affect the reaction can be used, preferably tetrahydrofuran/water, dioxane/water or acetone/water. The reaction is usually carried out at a temperature of 0-30°C for from 30 min. to 24 hours. This method using a base is preferably used when R 2 is methyl, ethyl, etc.

5) Reduction with reducing agents (different from catalytic reduction)

In this reaction, the group COOR 2 is converted to the group COOH by reduction in an inactive solvent, e.g. by the zinc-acid method. For this reaction, e.g. solvents such as acetone, water, dioxane, tetrahydrofuran, ethanol, acetonitrile, 1,1-dimethylformamide and acetic acid are used, alone or in combination. As acid, hydrochloric acid and acetic acid are suitable. The reaction is carried out at a temperature of 0-100°C, preferably 0-40°C, for 1-10 hours. The amount of zinc used for the reaction is usually 1-10 molar equivalents. This method is preferably used when R 2 is 2,2,2-trichloroethy1 etc.

Compound V, which is used as starting material, and methods for its production are specified in DE-OS no.

2911786. A representative example of the method steps is set forth below.

Furthermore, cephalosporins with a halogen substituent in the 3-position are prepared in the following reaction steps.

(G.V. Kaiser et al., "J.O. Chem", 35, 2430 (1970), R.R. Chauvette et al., "J. Am. Chem. Soc", 96, 4986 (1974)).

As can be seen from the above, the method according to the invention is carried out in four steps, i.e. 1) addition of thiol or selenol, 2). oxidation, 3) halogenation and 4) elimination; and if necessary additional steps such as 5) conversion of an azido- or phthaloyl-amino group to an amino group in the cis-position and/or 6) conversion of a protected carboxyl group to a carboxyl group in the 2-way Ilingen, using a 3H- carbacephem compound (eg, compound V) as starting material. Thus, hitherto unknown and useful 3-halogen-substituted carbacefem structures are obtained by the present method with hitherto unknown methods, which are completely different from the known methods for producing cephalosporin analogues.

The following examples illustrate the invention.

Example 1

Preparation of (-)-cis-7-azido-chloro-2-t-butyloxycarbonyl 1-1-azabicyclo-[4,2,0]-oct-2-en-8-one:

The above compound is produced by the following processes a), b), c) and d). Cis means stereo-isomerism at the 3- or 4-position of the 2-azetidinone ring or the 6- or 7-position of the 1-azabicyclo-[4,2,0]-octane ring, and this designation is also used in the following.

a) Preparation of (-)-cis-7-azido-3-phenylthio-2-t-butyl-oxycarbonyl-1-azabicyclo-[4,2,0]-octan-8-one:

528 mg (2 m mol) (-)-cis-7-azido-2-t-butyloxycarbonyl-1-azabicyclo-[4,2,0]-oct-2-en-8-one are dissolved in 15 ml of aqueous benzene . 0.2 ml (2 m mol) of thiophenol and 0.2 ml (2 m mol) of piperidine are then added to the solution and the mixture is stirred at room temperature for 2 hours. After the reaction, the reaction mixture is washed with 10% citric acid and saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The solvent is distilled off under reduced pressure and the resulting oily residue is purified by silica gel column chromatography (silica gel: "Wako-gel" R , C-200, (the same silica gel is used in the following), eluent: Ethyl acetate/n-hexane in the volume ratio 1: 4 (the same is used in the following) whereby 720 mg (yield 96.3%) of the desired compound with the following physical properties are obtained.

Melting point^77.5-78.0°C

IR(KBr)v<cm>: 2110, 1765, 17451

max.

NMR(CDCl3)6(ppm): 7.28-7.60(5H,m), 4.78(1H,d, J=5 Hz),

■4.33(1H,s), 3.78-3.98(1H,m), 3.81(1H,s), 1.50-2.34(4H,m)

l,42(9H,s)

b) preparation of (-)-cis-7-azido-3-phenylsulfinyl-2-t-butyloxycarbonyl-1-1-azabicyclo-[4,2,0]-octan-8-one:

480 mg (1.28 m mol) of the 3-phenylthio compound prepared in step a) is dissolved in 50 ml of anhydrous chloroform. Then 240 mg (1.41 m mol) of m-chloroperbenzoic acid are added to the solution under ice cooling. The reaction is carried out with stirring for 30 min. and then the reaction mixture is washed with saturated aqueous sodium bicarbonate solution

and saturated aqueous sodium chloride solution and dried over • aqueous sodium sulfate. The solvent is distilled off under reduced pressure whereby 500 mg (99.9%) of the desired compound with the following physical properties is obtained.

Melting point: 95.5-96.5°C

cm

IR(KBr)vmax 2120, 2100, 1780, 1735, 1035 NMR(CDC13)6(ppm): 7.55-7.91(5H,m), 4.87(1H,d, J=4.6Hz) ,

4.05(1H,s), 3.90-4.10(1H,m), 3.10(1H,s), 1.70-2.84(4H,m), 1.30(9H, s)

c) Preparation of (-)-cis-7-azido-3-chloro-3-phenylsulfinyl-2-t-butyloxycarbonyl-1-azabicyclo-[4,2,0]-octan-8-one: 109 mg of the The sulfoxide compound produced in step b) is dissolved in 1 ml of methylene chloride. Then 23.5 mg (0.42 m mol) of calcium oxide is added to the solution. Next, 27 yl ( (.34 m mol) of sulfinyl chloride is added to the mixture under ice-cooling. The reaction is carried out with stirring under ice-cooling for 1 hour. The reaction mixture is then washed with 10% citric acid, saturated aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfonate. The solvent is distilled off under reduced pressure and the resulting oily residue is purified by silica gel chromatography (5 g silica gel, eluent: ethyl acetate/h-hexane in the volume ratio 1:5), whereby 66.5 mg (56.1%) of the desired compound is obtained as an oily product with the following physical properties IR(CHCL3) %ax : 2120, 1770, 1735, 1055 NMR(CDC13)6(ppm): 7.53-8.00(5H,m), 4.90( lH,d, J=5Hz), 4.43(1H,s), 4.15-4.35(1H,m), 1.83-2.85 (4H,m), 1.38 (9H,s) d) Preparation of (-) -cis-7-azido-3-chloro-2-t-butyloxy- • carbonyl-1-azabicyclo-[4,2,0]-oct-2-en-8-one:

1.3 g (3.06 m mol) of the chloro-3-phenylsulfinyl compound obtained in the same way as described in step c) is dissolved in 100 ml of carbon tetrachloride and the solution is heated under reflux for 6 hours.

After completion of the reaction, the solvent is distilled off under reduced pressure and the residue is purified by silica gel chromatography (100 g of silica gel, eluent: ethyl acetate/n-hexane in the volume ratio 1:5), whereby 596 mg (65.2%) of the desired compound with the following physical properties. Melting point: 96-97°C

-1

IR(KBr)v^ks: 2120, 1765, 1735, 1630

PMR(CDCl 3 ) 6 (ppm) : 4.93(1H,d, J=5H z) , 3 , 7 2-3 , 9 2 (1H , m) ,

2.56-2.70(2H,m), 1.86-2.32(2H,m),

l,55(9H,s)

CMR (CDCl 3 ) 6 (ppm): 127.7, 125.0

Example 2

Preparation of (-)-cis-7-amino-3-chloro-2-t-butyloxycarbonyl-1-azabicyclo-[4,2,-0]-oct-2-3n-8-one:

350 mg (1.17 m mol) of that in Example 1, step d). prepared azido compound is dissolved in 20 ml of ethanol and 1.2 ml of IN hydrochloric acid and 70 mg of 10% palladium carbon are added to the solution. Hydrogen gas is introduced into the mixture at room temperature and atmospheric pressure for 3 hours. The palladium carbon is then filtered off and the filtrate is concentrated under reduced pressure. The resulting solid is dissolved in water and the solution is washed with ether. The aqueous layer is made slightly alkaline by the addition of sodium hydrogen carbonate and then extracted with ethyl acetate. The ethyl acetate layer is washed with saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The solvent is distilled off completely under reduced pressure whereby 218.4 mg (68.4%) of the desired compound is obtained as a powder with the following physical properties. Melting point: 102.5-104.5°C

-1

IR(KBr)v™ks: 1770, 1720, 1620

max.

NMR(CDCl 3 ) 6 (ppm) : 4 , 4 3 (1H , d, J=5Hz) , 3 , 5 2-3 , 9 0 (1H , m) ,

2.52-2.72(2H,m), 2.22(2H,br),

1.82-2.17(2H,m), 1.55(9H,s)

Example 3

Preparation of the trifluoroacetate of (-)-cis-7-amino-3-chloro-1.azabicyclo-[4,2,0]-oct-2-en-8-one-2-carboxylic acid: 1 ml of trifluoroacetic acid is added to 102 .2 mg (0.31 m mol) of the amino ester compound prepared in example 2 under ice cooling and the mixture is stirred at room temperature for 30 min.

The solvent is distilled off under reduced pressure and ether is added to the resulting oily residue to make it powdery. The residue is collected by suction filtration and dried in vacuum whereby 75.5 mg (60.9%) of the desired compound with the following physical properties is obtained.

Melting point: 208-220°C (decomp.)

-1

IR(KBr) v^ks : 1795, 1630

Example 4

Preparation of (-)-cis-7-azido-3-chloro-1-azabi-cyclo-[4,2,0]-oct-2-ene-8-one-2-carboxylic acid:

2 ml of trifluoroacetic acid are added to 83.9 mg

(0.28 m mol) of the azido ester compound prepared in example 1, step d), and the mixture is stirred at room temperature for 30 min. The solution is distilled off under reduced pressure and ether is added to the oily residue to make it powdery. The residue is collected by suction filtration and dried in vacuum and this gives 25.0 mg (36.7%) of the desired compound with the following physical properties.

Melting point: 147 , 5--148 , 5°C

-1

possibly

.IR(KBr)v^aks : 2130, 1770, 1715, 1605

NMR(CD3OD) 6 (ppm) : 5 , 16 (1H , d, J=5 Hz), 3 , 8 2-4 , 0 2 (1H , m) ,

2.63-2.77(2H,m), 1.73-2.21(2H,m)

In the following reference examples, methods for the production of desired end products from the intermediate products according to the invention are illustrated

■ Reference example 1

Preparation of (+ )-cis-7-[ 2-(•2-chloroacetamido-thiazol-4-yl)-2-syn-methoxyiminoacetylamino]-3-chloro-1-azabicyclo-[4,2,0]-oct -2-en-8-one-2-carboxylic acid:

122.6 mg (0.44 m mol) of 2-(2-chloroacetamidothiazol-4-yl)-2-syn-methoxyimineacetic acid are dissolved in 2.5 ml of anhydrous methylene chloride. Then 68 µl (0.49 m mol) of triethylamine are dissolved therein, and 92.0 mg. (0.44 m mol) phosphorus pentachloride is added while cooling in an ice/salt bath. The mixture is stirred at this temperature for 1 hour, after which 5 ml of n-

hexane is added. The mixture is stirred under ice cooling for a further 15 min. A separated oily material is obtained by decanting off n-hexane. The oily material is dissolved in 4 ml of tetrahydrofuran to produce an acid chloride solution. In a separate container, dissolve 121.7 mg (0.37 m mol) of the trifluoroacetate of ( + )-7-amino-3-chloro-1-azabicyclo-[4,2,0]-oct-2-ene- 8-one-2-carboxylic acid, prepared in the same manner as described in Example 3, in 5 ml of 50% aqueous tetrahydrofuran and 0.2 ml (1.47 mmol) of triethylamine. The above-prepared acid chloride solution in tetrahydrofuran is added to this solution with stirring and ice-cooling. The mixture is stirred at this temperature for 1 hour and set

to pH 3 with a N hydrochloric acid. Water is then added and the mixture is extracted with ethyl acetate. The extract is washed with saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. The solvent is distilled off to obtain 53.9 mg (30.5%) of the desired compound as a powder with the following physical properties.

-1

cm

lR(KBr)v^aks : 1770, 1680, 1555, 1045 .

NMR(DMSO-d66 (ppm) : 9.38(1H,&, J-(Hz), 7.37(1H,2),

5.45(lH,q, J=5, 8Hz), 4.35(2H,s)

Reference example 2

Preparation of (+)-cis-7-[2-(2-aminothiazol-4-yl)-2-syn-methoxyiminoacetamido]-3-chloro-1-azabicyclo-[4,2,0]-oct-2- en-8-one carboxylic acid:

51.2 mg (0.11 m mol) of the chloroacetamido compound prepared in reference example 1 is dissolved in 1 ml of dimethylacetamide, and 16.3 mg (0.22 m mol) of thiourea is added. The reaction is carried out by stirring the mixture at room temperature for 14 hours.

Then 7 ml of ether is added to the reaction mixture and stirring is continued for a further 10 min. A separated oily material is obtained by decanting off the ether. The oily material is dissolved in a small amount of dimethylsulfoxide, absorbed on 10 ml of "HP-10" resin in a column and eluted with dimethylsulfoxide. The eluate is treated twice with "HP-10" resin (eluent: water/methanol (the amount of methanol is gradually increased, and the final elution is carried out with water/methanol in the volume ratio 1:1)), thereby obtaining 15, 2 mg (35.4%) of the desired compound with the following physical properties. Melting point: 185.0-188.0°C (decomp.)

-1

IR(KBr)v^ks: 1765, 1670, 1630, 1540, 1040

NMR(DMSO-dg)6(ppm): 9.28(1H,d J=8.8 Hz), 7.17(2H,s),

6.75(lH,s), 5.44 (lH,q, J=5.3,

(.8 Hz.), 3.84(3H,s), 1, 24-2 , 52 (4H,m)

The antibacterial activity of the compound prepared in reference example 2 is indicated in the following table. The activity is determined by the heart infusion agar dilution method (pH 7.2) and Cefazolin is used for comparison.

Claims (9)

1. Cephalosporin-analogue 3-halocarbacefem compounds are characterized by the general formula wherein X means amino,' azido or protected amino, Hal means halogen, and R means carboxyl or protected carboxyl and salts thereof. .2. Compounds according to claim 1, characterized in that the hydrogen atoms in the 6- and 7-positions have cis configuration. 3. Compound according to claim 1 or 2, characterized in that the amih deprotecting group is selected from phthaloyl, t-butyloxycarbonyl, 2,2,2-trichloroethyloxycarbonyl, 2,2,2-trifluoroethyloxycarbonyl, benzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p-methoxybenzyl -oxycarbony1, phenylacetyl, phenoxyacety1, benzylidene and salicylidene. 4. Compound according to any one of the preceding claims, characterized in that Hal is chlorine. 5. Compound according to any of the preceding claims, characterized in that the protective carboxyl group is represented by the general formula -COOR, where R means alkyl with 1-5 carbon atoms, halogenated alkyl with 1-5 carbon atoms , arylmethyl with 7-20 carbon atoms, arylmethyl with 8-20 carbon atoms and optionally substituted with methoxy or nitro on the phenyl ring, substituted silyl or a group which is easily removed enzymatically or non-enzymatically in vivo. 6. Compound according to claim 5, characterized in that R 2 is t-butyl, benzyl, diphenylmethyl, triphenylmethyl, p-nitrophenylmethyl or p-methoxyphenyl. 7. Process for the preparation of cephalosporin-analogue 3-halocarbacefem compounds according to claim 1, with the general formula: 2 where Hal means halogen and R means carboxyl or -COOR, where R 2 means optionally halogenated alkyl with 1-5 carbon atoms, arylmethyl with 7-20 carbon atoms, arylmethyl with 8.20 carbon atoms and substituted with methoxy or nitro on the phenyl ring, substituted with silyl or a group that is easily removed enzymatically or non-enzymatically in vivo, characterized by: d that the X"*~a group in a compound of the formula: let where X means azido- or phthaloylamino and Hal and R have the meaning given above, is converted into an amino group. 8. Process for the preparation of cephalosporin analogue 3-halocarbacefem compounds according to claim 1 with the general formula: where X"^" means azido or protected amino, where the protecting group is selected from phthaloyl, t-butyloxycarbonyl, 2,2,2-trichloroethyloxycarbonyl, 2,2,2-trifluoroethyloxycarbonyl, benzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p- methoxybenzyl <p> oxycarbonyl, phenylacety1, phenoxyacety1,. benzylideneoxalisylidene, Hal means halogen and R means carboxyl or -COOR <2> , where R <2> means optionally halogenated alkyl with 1-5 carbon atoms, arylmethyl with 7-20 carbon atoms, arylmethyl with 8-20 carbon atoms and substituted methoxy or nitro on the phenyl ring, substituted xylyl or a group that is easily removed enzymatically or non-enzymatically in vivo, characterized in that R <1-> Y-OH is removed from a compound of the formula: where Y means a sulfur or a selenium atom, R"*" means optionally substituted alkyl, aralkyl, aryl or a heterocyclic group and X"<*> ", Hal and R have the above meaning. 9. Process for the preparation of cephalosporin analogue 3-halocarbacefem compounds according to claim 1, with the general formula: where X means amino-, azido or protected amino, where the protecting group is selected from phthaloyl, t-butyloxycarbonyl, 2,2,2-trichloroethyloxycarbonyl, 2,2,2-trifluoroethyloxycarbonyl, benzyloxycarbonyl, p-nitrobenzyloxycarbonyl, p- methoxybenzyloxycarbony1, phenylacetyl, phenoxyacety1, benzylidene and salicylidene, and Hal means halogen, characterized in that the group -COOR"", in a compound of the formula: where R 2 means optionally halogenated alkyl with 1-5 carbon atoms, arylmethyl with 7-20 carbon atoms, arylmethyl with 8-20 carbon atoms and substituted with methoxy or nitro on the phenyl ring, substituted silyl or a group that is easily removed enzymatically or non-enzymatically in vivo, and Hal has the above meaning, is converted to -COOH.