VINYLPYRROLIDINON CEPHALOSPORIN DERIVATIVES

CA 02220188 1997-11-04 F.HOFFMANN-LA ROCHE AG, CH-4070 Basle/Switzerland RAN 4410/257 Vinylpyrrolidinon Cephalosporin Derivatives The present invention relates to cephalosporin derivatives of the general formula H RI"(X)s'(CR4R5)m" (CH2 )n O N-R2 O COR3 O where R1 R4, R5 X n m S R2 is halogen, lower alkyl, phenyl, benzyl, styryl, naphthyl or heterocyclyl; the lower alkyl, phenyl, benzyl, styryl, naphthyl and heterocyclyl being optionally substituted by at least one of halogen, hydroxy, optionally substituted lower alkyl, optionally substituted lower alkoxy, optionally substituted phenyl, amino, lower alkylamino, di-lower alkyl mino, carboxy, lower alkylcarboxy, carbamoyl or lower alkylcarbamoyl; independently are hydrogen, lower alkyl or phenyl; is S, O, NH or CH2 ; is 0, 1 or 2; is 0 or 1; is 0 or 1; is hydrogen, hydroxy, -CH2 -CONHR6, lower alkyl-Qr, cycloalkyl-Qr, lower alkoxy, lower alkenyl, cycloalkenyl-Qr, lower alkynyl, aralkyl-Qr, aryl-Qr, aryloxy, aralkoxy, a heterocyclic ring or a heterocyclyl-Qr, the lower alkyl, cycloKj/10.9.97 CA 02220188 1997-11-04 Q r R alkyl, lower alkoxy, lower alkenyl, cycloalkenyl, lower alkynyl, aralkyl, aryl, aryloxy, aralkoxy and the heterocyclic ring may be substituted with at least one group selected from carboxy, amino, nitro, cyano, -S02 NHR6, optionally fluoro substituted lower alkyl, lower alkoxy, hydroxy, halogen, -CONR6R7, -CH2CONR6R7, -N(R7)COOR8, R7CO-, R7OCO-, R7COO-, -C(R7R9)CO2 R8, -C(R7R9)CONR7R10, wherein R6 is hydrogen, lower alkyl, cycloalkyl or aryl; R7 and R9 are independently hydrogen or lower alkyl; R8 is hydrogen, lower alkyl, lower alkenyl or a carboxylic acid protecting group; and R10 is hydrogen, œe-hydroxy-alkyl, phenyl, naphthyl or heterocyclyl, the phenyl, naphthyl or heterocyclyl being unsubstituted or substituted with at least one of the groups of optionally protected hydroxy, halogen, optionally substituted lower alkyl or oe-hydroxyalkyl, optionally substituted lower alkoxy and/or cyano, or R7 and R10 form together group of formula is -CHR-, -COor -SO2-; is 0 or 1; is hydrogen or lower alkyl; and is hydroxy, -O-, lower-alkoxy, or -OM and M represents an alkali metal; as well as readily hydrolyzable esters thereof, pharmaceutically acceptable salts of said compounds and hydrates of the compounds of formula I and of their esters and salts.

The compounds of the present formula I are useful in the treatment of infectious diseases caused by grain-positive bacteria, especially infectious diseases caused by sensitive and resistant staphylococci, pneumococci, enterococci and the like.

In above compounds of formula I the substituent in position 3 can be present CA 02220188 1997-11-04 in the E-form:

or in the Z-form:

N-- R2 O O. R2 I CH2)n Fo Ia Compounds of formula I i.e. wherein the substituent in position 3 is in the E-form are generally preferred.

In a particular embodiment of the compounds of formula I n is 1.

The term "halogen" or "halo" used herein refers to all four forms, that is chlorine or chloro; bromine or bromo; iodine or iodo; and fluorine or fluoro, unless specified otherwise.

As used herein, the terms "alkyl" and "lower alkyr' refer to both straight and branched chain saturated hydrocarbon groups having 1 to 8, and preferably 1 to 4, carbon atoms, for example, methyl, ethyl, n-propyl, isopropyl, tbutyl and the like.

By the term "optionally. substituted lower alkyl" is meant a "lower alkyl" moiety as defined above substituted by, for example, halogen, amino, hydroxy, cyano, carboxy etc., such as carboxymethyl, 2-fluoroacetyl, trifluoromethyl, 2,2,2-trifluoroethyl, 2-chloroethyl, 2-hydroxyethyl and like.

As used herein, the term "lower alkoxy" refers to a straight or branched chain hydrocarbonoxy group wherein the "alkyr' portion is a lower alkyl group as defined above. Examples include methoxy, ethoxy, n-propoxy and the like. The "alkyl" portion may be substituted as defined above.

As used herein, "alkenyl" and "lower alkenyl" refer to unsubstituted or substituted hydrocarbon chain radical having from 2 to 8 carbon atoms, preferably from 2 to 4 carbon atoms, and having at least one olefinic double bond, e.g. allyl, vinyl etc.

As used herein, "lower alkynyr' refers to unsubstituted or substituted hydrocarbon chain radical having from 2 to 8 carbon atoms, preferably 2 to 4 carbon atoms, and having at least one triple bond.

CA 02220188 1997-11-04 By the term "cycloalkyl" is meant a 3-7 membered saturated carbocyclic moiety, e.g., cyclopropyl, cyclobutyl, cyclohexyl, etc.

As used herein, "cycloalkenyr' refers to a carbocyclic ring radical having at least one olefinic double bond.

By the term "aryr' is meant a radical derived from an aromatic hydrocarbon by the elimination of one atom of hydrogen and can be substituted or unsubstituted. The aromatic hydrocarbon can be mononuclear or polynuclear. Examples of aryl of the mononuclear type include phenyl, tolyl, xylyl, mesityl, cumenyl, and the like. Examples of aryl of the polynuclear type include naphthyl, anthryl, phenanthryl, and the like. The aryl group can have at least one substituent selected from, as for example, halogen, hydroxy, cyano, carboxy, nitro, amino, lower alkyl, lower alkoxy, carbamoyl, such as in 2,4-difluorophenyl, 4-carboxyphenyl, 4-nitrophenyl, 4aminophenyl, 4-methoxyphenyl.

By the term "aralkyl" is meant an alkyl group containing an aryl group. It is a hydrocarbon group having both aromatic and aliphatic structures, that is, a hydrocarbon group in which a lower alkyl hydrogen atom is substituted by a monocyclic aryl group, e.g., phenyl, tolyl, etc.

As used herein, "aryloxy" is an oxygen radical having an aryl substituent (i.e., -O-aryl).

As used herein, "aralkoxy" is an oxygen radical having an aralkyl substituent.

As used herein, the term "lower alkylamino and i-lower alkylamlno" refers to mono and dialkyl_amino residues wherein lower alkyl is as defined above, for example methylamino, 2-ethylamino, -CH2 NHCH3 , -CH2CH2NHCH3,-CH2CH2N(CH3)2, N-methylamlno, N-ethylamino, N,Ndimethylamino, N,N-diethylamino and the like.

As used herein, "heter0cyclic ring" refers to an unsaturated or saturated, unsubstituted or substituted 4-, 5-, 6-, or 7-membered heterocyclic ring containing at least one hetero atom selected from the group consisting of oxygen, nitrogen, or sulfur. Exemplary heterocyclic rings include, but are not limited to, for example, the following groups: azetidinyl, pyridyl, pyrazinyl, piperidyl, piperidino, N-oxido-pyridyl, pyrimidyl, piperazinyl, pyrrolidinyl, pyridazinyl, N-oxide-pyridazinyl, pyrazolyl, tñazinyl, CA 02220188 1997-11-04 imidazolyl, thiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-triazolyl, 1,2,4triazolyl, 1H-tetrazolyl, 2H-tetrazolyl; furyl, 1H-azepinyl, thiophenyl, tetrahydrothiophenyl, isoxazolyl, isothiazolyl, oxazolidinyl, etc. Substituents for the heterocyclic ñng include, for example, lower alkyls such as methyl, ethyl, propyl, etc., lower alkoxys such as methoxy, ethoxy, etc., halogens such as fluorine, chlorine, bromine, etc., halogen substituted alkyls such as triíluoromethyl, trichloroethyl, etc., amino, mercapto, hydroxyl, carbamoyl, or carboxyl groups. A further substituent is oxo, such as in 2-oxo-oxazolidin3-yl, 1,1-dioxo-tetrahydrothiophen-3-yl. Further examples of substituted heterocycles are 6-methoxy-pyridin-3-yl, 5-methyl-isoxazol-3-yl, 1-methylpyridinium-2-yl,-3-yl,-4-yl, 1-carbamoylmethyl-pyridinium-2-yl, 1-carbamoylmethyl-pyridinium-3-yl, 1-carbamoylmethyl-pyridinium-2-yl,-3yl,-4-yl, 1-[N-(3-fluoro-4-hydroxy)phenyl]-carbamoylmethyl-pyridinium-2-yl.

By the term "substituted phenyl" is meant phenyl mono or disubstituted by halogen, optionally substituted lower alkyl, optionally protected hydroxy or amino, nitro or trifluoromethyl.

The term "optionally protected hydroxy " refers to hydroxy or hydroxy protected, for example with t-butyloxycarbonyl, trimethylsilyl, t-butyldimethylsilyl, tetrahydropyranyl, trifluoroacetyl, and the like, or refers to an ester group, for example, phosphate or sulfonate.

The term "optionally protected amino" refers to amino or amino protected with, for example, BOC [t-butoxycarbonyl; other name: (1,1dimethylethoxy)carbonyl], benzyloxycarbonyl and allyloxycarbonyl.

As used herein pharmaceutically acceptable salts useful in this invention include salts derived from metals, the ammonium salt, quaternary Ammonium salts derived from organic bases and amino acid salts. Examples of preferred metal salts are those derived from the alkali metals, for example, lithium (Li+), sodium (Na+) and potassium (K+).

Examples of quaternary ammonium salts derived from organic bases include tetramethylammonium (N+(CH3)4), tetraethylsmmonium (N+(CH2CH3)4), benzyltrimethylammonium (N+(C6H5CH2)(CH3)3), phenyltñethylammonium (N+(C6H5)(CH2 CH3)3), and the like, etc. Those salts derived from amines include salts with N-ethylpiperidine, procaine, dibenzylamine, N,N'-dibenzylethylenediamine, alkylamlnes or CA 02220188 1997-11-04 dialkylamines as well as salts with amino acids such as, for example, salts with arginine or lysine. Especially preferred are hydrochloñdes, sulfates, phosphates, lactates, mesylates or the inner salt.

The term "amino protecting groups" refers to protecting groups conventionally used to replace an acidic proton of an amino group. Examples of such groups are described in Green, T., Protective Groups in Organic Synthesis, Chapter 7, John Wiley and Sons, Inc. (1981), pp. 218-287, herein incorporated by reference. Preferably these examples include carbamates, e.g fluorenylmethyl, 2,2,2-trichloroethyl, 2-haloethyl, 2-(trimethylsilyl)ethyl, t-butyl, allyl, benzyl. Further protecting groups are 3,5-dimethoxybenzyl, pnitro-benzyl, diphenylmethy!, triphenylmethyl, benzyl, formyl, acetyl, trifluoroacetyl, chloro-acetyl, the cyclic imides of N-phthaloyl, Ntrimethylsilyl, N-benzenesulfonyl, N-toluenesulfonyl, N-p-methylbenzylsulfonyl. Preferred is BOC [t-butoxycarbonyl, other name (1,1-dimethylethoxy)carbonyl], benzyloxycarbonyl and allyloxycarbonyl.

The term "carboxylic acid protecting group" refers to protecting groups conventionally used to replace the acidic proton of a carboxylic acid.

Examples of such groups are described in Greene, T., Protective Groups in Organic Synthesis, Chapter 5, pp. 152-192 (John Wiley and Sons, Inc. 1981), incorporated herein by reference. Preferably these examples include methoxymethyl, methylthiomethyl, 2,2,2-tfichloroethyl, 2-haloethyl, 2- (trimethylsilyl)ethyl, t-butyl, allyl, benzyl, tñphenylmethyl (tñtyl), benzhydryl, p-nitrobenzyl, p-methoxybenzyl, trimethylsilyl, tñethylsilyl, tbutyldimethylsilyl, i-propyl-dimethylsilyl. Preferred are benzyhydryl, t-butyl, p-nitrobenzyl, p-methoxybenzyl and aUyl.

The term "hydroxy protecting group" refers to protecting groups as conventionally used in the art such as trimethylsilyl, t-butyl-dimethylsilyl, dimethylphenylsilyl, triphenylmethyl, lower alkanoyl, acetyl, tetrahydropyranyl, benzyl, p-nitrobenzyl or tbutyloxycarbonyl.

As readily hydrolyzable esters of the compounds of formula I there are to be understood compounds of formula I, the carboxy group(s) of which (for example, the 2-carboxy group) is/are present in the form of readily hydrolyzable ester groups. Examples of such esters, which can be of the conventional type, are the lower alkanoyloxy-alkyl esters (e.g., the acetoxymethyl, pivaloyloxymethyl, 1-acetoxyethyl and 1-pivaloyloxyethyl ester), the lower alkoxycarbonyloxyalkyl esters (e.g., the methoxycarbonylCA 02220188 1997-11-04 oxymethyl, 1-ethoxycarbonyloxyethyl and 1-isopropoxycarbonyloxyethyl ester), the lactonyl esters (e.g., the phthalidyl and thiophthalidyl ester), the lower alkoxymethyl esters (e.g., the methoxymethyl ester) and the lower alkanoylaminomethyl esters (e.g., the acetamidomethyl ester). Other esters (e.g., the benzyl and cyanomethyl esters) can also be used. Other examples of such esters are the following: (2,2-dimethyl-l-oxopropoxy)methyl ester; 2-[(2methylpropoxy)carbonyl]-2-pentenyl ester; 1-[[(1-methylethoxy)carbonyl]oxy] ethyl ester; 1-(acetyloxy) ethyl ester; (5-methyl-2-oxo-l,3-dioxol-4-yl) methyl ester; 1-[[(cyclohexyloxy)carbonyl]oxy] ethyl ester; and 3,3-dimethyl-2-oxobutyl ester. It will be appreciated by those of ordinary skill in the art that the readily hydrolyzable esters of the compounds of the present invention can be formed at a free carboxy group of the compound.

Examples of salts of the compounds of formula I are defined under "pharmaceutically acceptable salts" above.

A preferred embodiment of the invention are compounds of formula I wherein R1 is selected from the groups phenyl, 2,4,5-trichlorophenyl, 3,4dichlorophenyl, 2,5-dichlorophenyl, 4-tñfluoromethylphenyl, 4-methoxyphenyl, 4-hydroxymethylphenyl, 3,4-dimethoxyphenyl, 4-methyl-l,2,4triazol-5-yl, 1-methyl-tetrazol-5-yl, pyrimidin-2-yl, optionally substituted pyridinium-l-yl, 2-yl, -3-yl or-4-yl, benzim dazol-2-yl, 2-benzthiazolyl, 4pyñdinyl, (2-amíno)-thiazol-4-yl, 2-naphthyl, benzyl and R2 is methylcyclopropyl, 2-, 3or 4-hydroxybenzyl, pyrrolidin-3-yl or a group of formula wherein Q1 r Rll (Q1)r 0 I is -CH2 - is 0 or 1; is hydrogen, lower alkyl, oe-hydroxy alkyl, benzyl or alkylheterocyclyl, the benzyl and the heterocyclyl group being unsubstituted or substituted with at least one of the groups cyano, carboxy or hydroxy; or is -CH2 CONR7RI°; wherein R7 and R10 are as defined above.

The compounds of formula I as well as their salts and readily hydrolyzable esters can be hydrated. The hydration can be effected in the CA 02220188 1997-11-04 course of the manufacturing process or can occur gradually as a result of hygroscopic properties of an initially anhydrous product.

Especially preferred compounds of formula I are (E)-(6R,7R)-3-[1-[1-[(4-Hydroxy-phenylcarbamoyl)-methyl]-pyridin-l-ium-4ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-[2-(2,4,5-trichlorophenylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate CI /NH-- -OH I O/'--%O C' (E)-(6R,7R)-3-[1-[ 1-[(4-Hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-[2-(1-benzothiazol-2-ylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate N H /tií--- N% O co o (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7- [2-(2,4,5-tñchloro-phenylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid ¢1 CI O H COOH (E)-(6R,7R)-7-[2-(Benzothiazol-2-ylsulfanyl)-acetylamino]-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid CA 02220188 1997-11-04 H CO2H O (E)-(6R,7R)-3-[1-[ 1-[(4-Hydroxy-phenylcarbamoyl)-methyl]-pyridin1-into-4ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-[2-(pyridin-4-ylsulfanyl)- acetylamlno]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate o OH (E)-(6R,7R)-3-[ 1-[1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1- ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-(2,4,5-trichlorophenylsulfanyl)-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate CI (LIGHTOH O F (E)-(6R,7R)-3-[1-[ 1-[(4-Hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-(2-phenylsulfanyl-acetylam uo)-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate H N O CA 02220188 1997-11-04 (E)-(6R,7R)-3-[1-[1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin-1inm-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-7-[2-(naphthalen-2-ylsulfanyl)-acetylamino]-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate __J c¢2 o (E)-(6R,7R)-7-[2-(3,4-Dichloro-phenylsulfanyl)-acetylamino]-3-[1-[1-[(3-fluoro4-hydroxy-phenyl carb amoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]ooE-2-ene-2carboxylate CI1 / HN-- OH 0 F (E)-(6R,7R)-3-[1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- [2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid ci COOH 0 (E)-(6R,7S)-3-[1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- [2-naphthalen-2-ylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0] oct-2-ene2-carboxylic acid C02H 0 CA 02220188 1997-11-04 1,5 Mixture of(E)-(6R,7R)-8-oxo-3-[(R)- and -[(S)-2-oxo-[1,3']Bipyrrolidinyl-3ylidenemethyl]-8-oxo-7-[2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino]-5thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid hydrochloride CI CI H CO2H • HCl (E)-(6R,7R)-8-Oxo-3-[(R)-2-oxo-[1,3']bipyrrolidinyl-3-ylidenemethyl]-7-(2phenylsulfanyl-acetylamino)-5-thia1-aza-bicyclo[4.2.0] oct-2-ene-2carboxylicacid hydrochloñde (1:1) s. ..

¢O2H O (E)-(6R,7R)-3-[1-[1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin-1ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-(2-phenylsulfanylacetylamlno)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate CO2 O (E)-(6R,7R)-7-[2-(2,5-DicMoro-phenylsulfanyl)-acetylam no]-3-[1-[(3-fluoro-4hydroxy-phenylcarbamoyl)-methyl]-pyridín1-ium-4-ylmethyl]-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate CI H / "" N+/'- O N S \ NH OH CO2 O CA 02220188 1997-11-04 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2- (naphthalen-l-ylsulfanyl)-acetylamino]-8-oxo-5-thía-l-aza-bicyclo[4.2.0]oct-2ene-2-carboxylic acid imidazole salt (1:1) C02H 0 The compounds of the present invention are useful as antibiotics having potent and broad antibacterial activity; especially against grain-positive organisms, e.g. methicillin-resistent staphylococci (MRSA).

The products in accordance with the invention can be used as medicaments, for example, in the form of pharmaceutical preparations for parenteral administration, and for this purpose are preferably made into preparations as lyophilisates or dry powders for dilution with customary agents, such as water or isotonic common salt solution.

Depending on the nature of the pharmacologically active compound the pharmaceutical preparations can contain the compound for the prevention and treatment of infectious diseases in mammals, human and non-human, a daily dosage of about 10 mg to about 4000 mg, especially about 50 mg to about 3000 mg, is usual, with those of ordinary skill in the art appreciating that the dosage will depend also upon the age, conditions of the mammals, and the kind of diseases being prevented or treated. The daily dosage can be administered in a single dose or can be divided over several doses. An average single dose of about 50 mg, 100 mg, 250 mg, 500 mg, 1000 mg, and 2000 mg can be contemplated.

Representative compounds (A to F, below) of the present invention were tested. In vitro activity was determined by minimum inhibitory concentration in a microorganism spectrum by the agar dilution method in Mueller Hinton agar, inoculum = 104 CFU/spot.

A: OE)-(6R,7R)-3-[1-[ 1-[(4-Hydroxy-phenylcarb_amoyl)-methyl]-pyridinl-Jure4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-[2-(2,4,5-trichlorophenylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate B: (E)-(6R,7R)-3-[1-[1-[(4-Hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo7-[2-( 1-benzothiazolCA 02220188 1997-11-04 D:

E:

G:

H:

2.,5 2-ylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate C: (E)-(6R,7R)-7-[2-(Benzothiazol-2-ylsulfanyl)-acetylamino]-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo- [4.2.0]oct-2-ene-2-carboxylic acid (E)-(6R,7R)-3-[ 1-[1-[(4-Hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-[2-(pyridin-4-ylsulfanyl)-acetylamino]-5:thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate (E)-(6R,7R)-3-[1-[1-[(3-Fluoro-4-hydroxy-phenylcarbAmoyl)-methyl]- pyridin-l-ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-7-[2- (naphthalen-2-ylsulfanyl)-acetylamino]-8-oxo-5-thia1-aza-bicyclo- [4.2.0]oct-2-ene-2-carboxylate F: (E)-(6R,7R)-3-[1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8oxo-7-[2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino]-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (E)-(6R,7S)-3-[1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8oxo-7-[2-naphthalen-2-ylsulfanyl)-acetylamino]-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid Mixture of (E)-(6R,7R)-8-oxo-3-[(R)- and -[(S)-2-oxo-[1,3']Bipyrrolidinyl-3ylidenemethyl]-8-oxo-7-[2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino]-5thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid hydrochloride I: (E)-(6R,7R)-3-[ 1-[ 1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl]- pyridin-l-ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-(2phenylsulfanyl-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate In vitro activity against sensitive and resistant S. aureus MIC g/ml] MIC S.aureus 6538 (MSSA) MIC S. aureus 270A (MRSA) A <0.O6 B 0.12 C 0.25 D E 0.5 0.25 4 1 F 0.12 G 0.12 H < O.O6 Agar dilution method on Mueller-Hinton agar, inoculum: 104 CFU/spot I _< 0.06 CA 02220188 1997-11-04 The compounds of the formula I in accordance with the invention as well as their pharmaceutical acceptable salts, hydrates, or readily hydrolyzable esters can be manufactured in accordance with the invention by (a) treating a compound having the formula II RfHN« II COOR g 0 in which R2 and n are defined above; Rf is hydrogen or trimethylsilyl; and Rg is hydrogen, benzhydryl, p-methoxybenzyl, t-butyl, trimethylsilyl or allyl or salt thereof, with a carboxylic acid of the general formula III R1 -(X)s-(CR4R5) m ,, ,Y III in which RI, X, s, R4, R5 and m are as defined above and Y is -OH, or a reactive functional derivative thereof wherein Y is, for example a halogen as chloride or bromide, or 1-imldazolyl, 2-mercaptobenzotriazolyl, 1-hydroxbenzotriazolyl or pivaloyloxy, or an activating agent as HBTU (ortho-Benzotñazol1-yl-N,N,N',N'-tetramethyluroniumhexafluorophosphat), DCC (N,N'-dicylohexylcarbodiimid), CDI (1,1'- carbonyl-di]m dazole), CDT (1,1'-carbonyl-l,2,4-ditriazole) or thionylchloride and the like.

(b) for compounds of formula I wherein X is S, 0 or NH, by treating a compound having the formula IV H HaI-(CR4R5)m N (CH2 )n 002R g 0 I V wherein R4, R5, m, n, R2 and Rg are as defined above and Hal stands for halogen as bromine or chlorine or iodide preferably bromine, CA 02220188 1997-11-04 with an appropriate thiol or thiolate or an appropriate alcohol or alcoholate or an appropriate amine in the presence of a base, (c) for the manufacture of a readily hydrolysable ester of a compound of formula I subjecting a carboxylic acid of formula I to a corresponding esteñfication or (d) for the manufacture of salt or hydrates of a compound of formula I or hydrates of said salts converting a compound of formula I into a salt or hydrate or into a hydrate of said salts.

The reaction of a compound of formula II prepared according to embodiment (a) with a compound of formula III, or a reactive derivative thereof can be carried out in a manner known per se. The carboxy groups in compounds of formula II (carboxy group in position 2 and/or carboxy groups optionally present in R2) in compounds of formula III (carboxy groups optionally present in R1) can be protected intermediatly or in situ, for exemple, by esterification to form readily cleavable esters such as a silyl ester (e.g. trimethylsilylester), a p-methoxy-benzylester or benzhydryl ester.

Furthermore the amino groups present in compounds of formula II (in position 7 and/or optionally present in R2) and/or optionally present in R1 of compounds of formula III can be protected, for example, with protecting groups which are cleavable by acid hydrolysis (e.g. the t-butoxycarbonyl or triphenylmethyl groups), by basic hydrolysis (e.g. the trifluoroacetyl group), by hydrazinolysis (e.g. the phthalimido group) or by catalytic cleavage in presence of Pd (the allyloxycarbonyl group). Preferred protecting groups are the t-butyloxy-carbonyl, allyloxycarbonyl, the chloroacetyl, bromoacetyl and iodoacetyl groups, especially the chloroacetyl group. These last-mentioned protecting groups can be cleaved off by treatment with thiourea. Another preferred protecting group is phenylacetyl which can be cleaved off by treatment with PC15 or enzymatic.

The 7-amino group in compounds II can be protected in situ, for example, by a silyl protecting group such as the trimethylsilyl group.

In reacting a 7-amino compound of formula II with a carboxylic acid of formula III or a reactive functional derivative thereof, for example, a free carboxylic acid can be reacted with an aforementioned ester of a compound of formula II in the presence of a carbodiimide such as dicyclohexylcarboCA 02220188 1997-11-04 diimide in an inert solvent such as ethyl acetate, acetonitrile, dioxane, chloroform, methylene chloride, benzene, dimethylformamide or dimethylacetamide, and subsequently the ester group can be cleaved off.

Prepared according to another embodiment, a salt of an acid of formula II (e.g. a tñalkylammonium sait such as the triethylamrnonium salt) is reacted with a reactive ñmctional derivative of a carboxylic acid of formula III in an inert solvent (e.g. dimethylformamide, dimethylacetamide, dimethylsulfoxide and the like).

The reaction of a 7-amino compound of formula II with the carboxylic acid of formula III or a reactive derivative thereof can conveniently be carried out at a temperature between about -40°C and +60°C, e.g. at room temperature.

Embodiment (b) of the process of the present invention involves treating a compound of formula IV with an appropriate thiol or thiolate or an appropriate alcohol or alcoholate or an appropriate amine in presence of a base, for example, tñalkylamine such as trimethylamine, triethylamin sodium bicarbonate, DBU (1,8-diazabicyclo[5,4,0]undec-7-en(1,5-5) to form the corresponding thioether, ether or amine. Optionally present amino, hydroxy or carboxyl groups can be intermediatly protected with groups as described above.

Deprotection (removal) of protected amino, hydroxy or carboxylic groups present in a compound of formulae II, III and IV can be carried out as follows:

Removal of amino urotectine arrouus Possible amino-protecting groups are those employed in peptide chemistry, such as the protecting groups mentioned above. Preferably these examples include carbamatès, e.g. fluorenylmethyl, 2,2,2-tñchloroethyl, tbutyl, triphenylmethyl, allyl, benzyl. Further protecting groups are p-nitrobenzyl, diphenylmethyl, triphenylmethyl, benzyl, formyl, trifluoroacetyl, chloro-acetyl, the cyclic imides of N-phthaloyl, N-trimethylsilyl, Nbenzenesulfonyl, N-toluenesulfonyl. Preferred is BOC [t-butoxycarbonyl; other name: (1,1-dimethylethoxy)carbonyl], benzyloxycarbonyl, allyloxyo carbonyl or tñmethylsilyl The amino protecting groups may be cleaved off by acid hydrolysis (e.g.

the t-butoxycarbonyl or tñphenylmethyl group), e.g. aqueous formic acid, CA 02220188 1997-11-04 trifluoroacetic acid or by basic hydrolysis (e.g. the trifluoroacetyl group). The chloroacetyl, bromoacetyl and iodoacetyl groups are cleaved off by treatment with thiourea. The tñmethylsilyl group is cleaved off by hydrolysis or alcoholysis.

Amino-protecting groups which are cleavable by acid hydrolysis are preferably removed with the aid of a lower alkanecarboxylic acid which may be halogenated. In particular, formic acid or trifluoroacetic acid is used. The reaction is carried out in the acid or in the presence of a co-solvent such as a halogenated lower alkane, e.g. methylene chloride. The acid hydrolysis is generally carried out at room temperature, although it can be carried out at a slightly higher or slightly lower temperature (e.g. a temperature in the range of about -30°C to +40°C). Protecting groups which are cleavable under basic conditions are generally hydrolyzed with dilute aqueous caustic alkali at 0°C to 30°C. The chloroacetyl, bromoacetyl and iodoacetyl protecting groups can be cleaved off using thiourea in acidic, neutral or alkaline medium at about 0°C-30°C. The phthalimido group can be cleaved off with hydrazine at -20°C to +50°C.

Removal of hydrox¥ protecting group Possible hydroxy protecting groups are such as are commonly known in the art, such as trimethylsilyl, t-butyl-dimethylsilyl, dimethylphenylsilyl, tñphenylmethyl, lower alkanoyl, acetyl, trifluoroacetyl, tetrahydropyranyl, benzyl, p-nitrobenzyl or t-butoxycarbonyl. These groups are removed in the presence of acidic solvents, weak organic acids or weak inorganic bases like s0di11m bicarbonate, respectively.

Removal of protecting a rouus at the carboxv function As carboxyl protecting groups one may utilize an ester form which can be easily converted into a free carboxyl group under mild conditions, for example, benzhydryl, t-butyl, p-nitrobenzyl, p-methoxybenzyl, allyl, etc.

CA 02220188 1997-11-04 These protecting groups may be removed as follows:

benzhydryl t-butyl p-nitrobenzyl p-methoxybenzyl allyl trifluoroacetic acid with anisol, phenol, cresol or tñethylsilane at about -40°C to room temperature; hydrogen with Pd/C in an alcohol such as ethanol or in tetrahydrofuran; BF3-etherate in acetic acid at about 0 to 50°C; formic acid or trifluoroacetic acid with or without anisol, phenol, cresol or tñethylsilane and a solvent such as dichloromethane at about -10°C to room temperature; sodium sulfide in acetone/water at about 0 to room temperature; or hydrogen with Pd/C in an alcohol such as ethanol or in tetrahydrofuran; formic acid at about 0 to 50°C; or trifluoroacetic acid and anisol, phenol or triethylsilane at about -40°C to room temperature; palladium(O) catalyzed transalkylation reaction in the presence of sodium or potassium salt of 2-ethyl hexanoic acid, see for example J. Org. Chem. 1982, 47, 587.

tñmethylsilyl by hydrolysis or alcoholysis at room temperature.

In order to manufacture a readily hydrolysable ester of the carboxylic acids of formula I in accordance with embodiment (c) of the process provided by the present invention, a carboxylic acid of formula I is preferably reacted with a corresponding halide, preferably an iodide, containing the desired ester group. The reaction can be accelerated with the aid of a base such as an alkali metal hydroxide, an alkali metal carbonate or an organic amine such as triethylamine. The esterification is preferably carried out in an inert organic solvent such as dimethylacetamide, hexamethylphosphoric acid triamlde, dimethyl sulfoxide or, especially, dimethylformamide. The reaction is preferably carried out at a temperature in the range of about 0-40°C.

ao The manufacture of the salts and hydrates of the compounds of formula I or the hydrates of said salts in accordance with embodiment (d) of the process provided by the present invention can be carried out in a manner known per se; for example, by reacting a carboxylic acid of formula I or a sait thereof with an equivalent amount of the desired base, conveniently in a solvent such as water or an organic solvent (e.g. ethanol, methanol, acetone and the like). Correspondingly, salt formation is brought about by the CA 02220188 1997-11-04 addition of an organic or inorganic salt or an acid. The temperature at which the sait formation is carried out is not critical. The sait formation is generally carried out at room temperature, but it can be carried out at a temperature slightly above or below room temperature, for example in the range of 0°C to +50°C.

The manufacture of the hydrates usually takes place automatically in the course of the manufacturing process or as a result of the hygroscopic properties of an initially anhydrous product. For the controlled manufacture of a hydrate, a completely or partially anhydrous carboxylic acid of formula I or sait thereof can be exposed to a moist atmosphere (e.g. at about +10°C to +40°C).

Exemplary of the process for obtaining products in accordance with the invention are the following reaction schemes 1 and 2 below.

Scheme 1, embodiment (a) RI-(X)s-(CR4R5)myY III o H RI"(x) s'(CR4R5)my N%r__I S-,,,I /..,,-(CH2)n " o I 002Rg 0 wherein X is -CH2 -, O, S or NH and the remaining symbols are as defined above.

Scheme 2, embodiment (b) R f NH* r. S- -(CH2 )n O/fl-- N-R2 C02Rg II 0 Hal-(CR4R 5)m . O Hal-( CR4RS)rn H .. S, ,.I(CH2)n Bror CI 0 I I / ! N-Ff C02Rg O R1 -X' H R l"(x) s'(CR4 RS)rn--,,r(N%. S,,h r,,,. (CH2 )n C02Rg 0 wherein X is O, S, NI{ and X' accordingly OH or O-, SH or Sor NH2 and the remaining substituent are as defined above.

CA 02220188 1997-11-04 Examples Met, h.od A E mpleA1 (E)-(6R,7R)-3-[ 1-[1-[(4-Hydroxy-phenylcarbamoyl)-methyl]-pyridin1-inm-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-[2-(2,4,5-tñchloro-phenylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate N S _/ To a solution of 68.4: mg (0.25 retool) (2,4,5-trichloro-phenylsulfanyl)-acetic acid in 3 ml N,N-dimethylacetamíde were added under stirring and Argon atmosphere 40.9 mg (0.25 retool) 1,1'-carbonyldiimidazole. After 30 rein, 136.4 mg (0.21 retool) (E)-(6R,7R)-7-amino-3-[1-[1-[(4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate trifluoroacetate was added in a single portion. After 3h the reaction mixture was poured on diethyl ether. The solid material was collected by filtration and triturated with ethyl acetate.

Yield: 112.0 mg (67.5%) beige powder IR(KBr): 1770, 1678, 1642 cm1 MS(ISP): 790.2 (M+) According to the procedure in example A1 the following additional compounds were prepared:

Example A2 (E)-(6R,7R)-3-[1-[1-[(4-Hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-[2-(1-benzothiazol-2ylsulfanyl)-acetylamlno]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate O CA 02220188 1997-11-04 With 70.0 mg (0.43 retool) 1,1'-carbonyldiimidazole, 96.0 mg (0.43 retool) (benzothiazol-2-ylsulfanyl)-acetic acid and 233.8 mg (0.36 retool) (E)-(6R,7R)- 7-amì no-3-[1-[ 1-[(4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylate trifluoroacetate in 4 ml N,N-dímethylacetamide.

Yield: 92.0 mg (34.4%) yellow powder IR(tçBr): 1769, 1679, 1643, 1625 cm"z MS(ISP): 743.3 (M+H÷) ET ample A3 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7- [2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid CI H _? Cl 0 Ç C02H O With 180.0 mg (1.11 L OI) l,l'-carbonyl imidazole, 301.4 mg (I.II retool) (2,4,5-tncllloro-phenylsulfanyl)-acetic acid and 323.2 mg (0.92 retool) (E)- (6R,7R)-7-amino-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid in 8 ml N,Ndimethylformamide.

Yield: 337.0 mg (60.4%) brown powder IR(KBr): 1773, 1668, 1621 cm"1 MS(ISP): 602.2 (M+H÷) ETample A4 (E)-(6R,7R)-7-[2-(Benzothiazol-2-ylsulfanyl)-acetylamino]-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid H O C02H Nçì> o With 220.0 mg (1.35 mmol) !,l'-carbonyldiimidazole, 304.1 mg (1.35 retool) (benzothiazol-2-ylsulfanyl)-acetic acid and 394.8 mg (1.13 retool) (E)-(6R,7R)- 7-_am no-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5CA 02220188 1997-11-04 1,5 thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid in 7 ml N,Ndimethylformam de.

Yield: 173.0 mg (27.5%) orange powder IR(KBr): 1772, 1665, 1623 cm1 MS(ISP): 557.1 (M+H÷) E -mpleA5 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7- (2-phenylsulfanyl-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid CO2H O With 167.0 mg (1.o3 retool) 1,1'-carbonyldiimidazole, 173.0 mg (1.03 retool) 2- (phenylthio)acetic acid and 300.0 mg (0.86 retool) (E)-(6R,7R)-7-amluo-3-(1cyclopropylmethyl-2-oxo-pyrrolidin-3-ylideneme/hyl)-8-oxo-5-/hia1-azabieyclo[4.2.0]oet-2-ene-2-carboxylie acid in 6 ml N,N-dimethylformamide.

Yield: 271.5 mg (63.1%) brown powder IR(IçBr): 1773, 1662, 1624 cm-1 MS(ISP): 500.3 (M+I-I÷) E mple A6 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7- (2-pyridin-4-ylsulfanyl-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid O H CO2H o With 111.0 mg (0.68 retool) 1,1'-carbonyldiimidazole, 116.0 mg (0.68 retool) (pyridin-4-ylsulfanyl)-acetic acid and 200.0 mg (0.57 mmol) ) (E)-(6R,7R)-7am uo-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid in 4 ml N,Ndimethylformamide.

Yield: 79.0 mg (27.5%) beige solid IR(KBr): 1769, 1667, 1624 MS(ISP): 501.1(M+H÷) CA 02220188 1997-11-04 ExAmple A7 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2- (naphthalen-2-ylsulfanyl)-acetyl mlno]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2ene-2-carboxylic acid C02H O With 167.0 mg (1.o3 mm01) 1,1'-carbonyldiimidazole, 225.0 mg (1.03 mmol (naph alen-2-ylsulfanyl)-acetie acid and 300.0 mg (0.86 retool) (E)-(6R,7R)-7amluo-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-azabicyclo[4.2.0]oet-2-ene-2-earboxylic add in 4 ml N,N-dimethylformamide.

Yield: 320.0 mg (62%) brown powder IR(KBr): 1769, 1662, 1623 cm1 MS(ISP): 550.2 (M+H÷) E mple AS (E)-(6R,7R)-7-[2-(2-Amino-thiazol-4-yl)-acetylamino]-3-[1-[1-[(4-hydroxyphenylcarbamoyl)-methyl]-pyridinl-ium-4-ylmethyl]-2-oxo-pyrrolidin-3ylidenemethyl]-8-oxo-5-thial-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate CO20 With 54.5 mg (0.34 retool) 1,1'-carbony]oEimJdazo]e, 53.0 mg (0.34 retool) (2aminotkdazole-4-y])-acetic acid and 182.0 mg (0.28 retool) (E)-(6R,7R)-7amino-3-[1-[ 1-[(4-hydroxy-pheny]carbamoyl)-methyl]-p din1-ium-4ylmet y]]-2-oxo-pÆolioEn-3-y]idenemethy]]-8-oxo-5-t]xìa-l-azabicyc]o[4.2.0]oct-2-ene-2-carboxy]ate tnfluoroacetate m 4 ml N,Ndimethylformamide. The resulting solid was purified by reversed phase chromatography (RP-18 LiChroPrep gel, water : acetonitñle = 9 : 1). The organic solvent was stripped, off at a rotary evaporator and the aqueous phase was freeze-dñed.

Yield: 49.0 mg (24.7%) beige lyophilisate IR(KBr): 1775, 1680, 1650 cm1 CA 02220188 1997-11-04 MS(ISP): 676.2 (M ÷) E ,-pleA9 (E)-(6R,7R)-7-[2-(2-Amino-thiazol-4-yl)-acetylamino]-3-(1-cyclopropylmethyl2-oho-pyrrolidìn-3-ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0] oct-2-ene-2carboxylic acid H S O CO2H O With 167.0 mg (1.03 m ol) 1,1'-carbony]dñmidazo]e, 163.0 mg (1.03 retool) (2amiuotMazole-4-yl)-acetic acid and 300.0 mg (0.86 m,no]) (E)-(6R,7R)-7amlno-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid in 6 ml N,N-dimethylformam de. The resulting solid was purified by reversed phase chromatography (RP-18 LiChroPrep gel, water : acetonitrile = 9 : 1). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 120.0 mg (28.6%) beige lyophilisate IR(KBr): 1769, 1664, 1620 cm;1 MS(ISP): 490.2 (M+H÷) mpleA10 (E)-(6R,7R)-3-[1-[1-[(4-Hydroxy-phenylcarbamoyl)-methyl]-pyridinl-Jure-4ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-[2-(pyridin-4-ylsulfanyl)- acetylam no]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate o With 70.0 mg (0.43 retool) 1,1'-carbonyldñmidazole, 72.8 mg (0.43 retool) (pyrìdin-4-ylsulfanyl)-acetic acid and 232.8 mg (0.36 retool) (E)-(6R,7R)-7am no-3-[1-[1-[(4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylate trifluoroacetate in 4 ml ml N,N-dimethylacetamide. The brown solid was purified by column chromatography on MCI gel (75-150 Mitsubishi Kasei Corporation) with a gradient of water : acetonitrile (1:0, 4:1, CA 02220188 1997-11-04 3:1, 2:1, 1:1). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 58.0 mg (30.0%) beige lyophilisate IR(KBr): 1772, 1670, 1625 cm"1 MS(ISP): 687.3 (M+H÷) ET-mple All (E)-(6R,7R)-3 - [ 1 -[ 1- [(4-Hydroxy-phenylcarhamoyl)-methyl] -pyridin1-ium-4ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-(2-phenylsulfanylacetylamlno)-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate o With 70.0 mg (0.43 retool) 1,1'-carbonyldiimidazole, 71.0 mg (0.43 retool) 2- (phenylthio)acetic acid and 188.4 mg (0.29 retool) (E)-(6R,7R)-7-amino-3-[1-[1- [(4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thìa1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate trifluoroacetate in 4 ml ml N,N-dimethylacet_amide. The resulting solid was suspended in 6 ml water : acetonitrile (1 : 1) and HC1 was added until all compound dissolved. After column chromatography on MCI gel (75-150 Mitsubishi Kasei Corporation) with a gradient of water :

acetonitril (1:0, 4:1, 3:1, 2:1, 1:1) the organic solvent was stripped offat a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 105.6 mg (64.4%) beige lyophilisate IR(KBr): 1770, 1680, 1643 cm1 MS(ISP): 686.3 (M+H÷) CA 02220188 1997-11-04 1,5 mpleA12 (E)-(6R,7R)-3-[1-[1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin-1ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-7-[2-(naphthalen-2-ylsulfanyl)-acetylamino]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate _/ o With 58.3 mg (0.36 retool) 1,1'-earbonyldiimidazole, 78.5 mg (0.36 rnmol) (naphthalen-2-ylsulfanyl)-aeetie and and 200.0 mg (0.30 retool) (E)-(6R,7R)-7amino-3-[1-[ 1-[(3-fluoro-4-hydroxy-phenylearbomoyl)-methyl]- pyridin-l-inm4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bieyelo- [4.2.0]oct-2-ene-2-carboxylate trifluoroacetate in 4 ml N,N-dimethylacetamide. The resulting solid was purified by column chromatography on MCI gel (75-150p, Mitsubishi Kasei Corporation) with a gradient of water : acetonitrile (1:0, 4:1, 3:1, 2:1, 1:1). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dñed.

Yield: 55.0 mg (24.0%) beige lyophilisate IR(KBr): 1770, 1680, 1650, 1628 cm"1 MS(ISP): 754.3 (M+H÷) i ample Al3 (E)-(6R,7R)-7-[2-(3,4-Dichloro-phenylsulfanyl)-acetylamino]-3-[ 1-[ 1-[(3-fluoro4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2-oxopyrrolidin3-ylidenemethyl]- 8oxo5-thia1 -aza-bicyclo [4.2.0] o ct-2ene2 - carboxylate CI / HNOH «\ // -- N 0 F H With 60.0 mg (0.36 retool) 1,.l'-earbonyldiimidazole, 85.0 mg (0.36 retool) [(3,4-dichlorophenyl)thio]acetic acid and 200.0 mg (0.30 retool) (E)-(6R,7R)-7amino-3-[1-[ 1-[(3-tluoro-4-hydroxy-phenylcarbamoyl)-methyl]- pyñdin-l-ium4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bieyelo- [4.2.0]oct-2-ene-2-carboxylate trifluoroacetate in 4 ml N,N-dimethylacetCA 02220188 1997-11-04 amide. The resulting solid was purified by column chromatography on MCI gel (75-150 t, Mitsubishi Kasei Corporation) with a gradient of water :

acetonitrile (1:0, 4:1, 3:1, 2:1, 1:1). The organic solvent was stripped offat a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 70.0 mg (30.2%) beige lyophilisate IR(KBr): 1772, 1680, 1642, 1617 cm1 MS(ISP): 772.3 (M+H÷) Example Al4 (E)-(6R,7R)-3-[ 1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- [2-(2,4,5-trichloro-phenylsulfanyl)-acetylnmino]-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid CI C02H 0 With 146.0 mg (0.90 retool) 1,1'-carbonyldiimldazole, 244.4 mg (0.90 retool) (2,4,5-triehloro-phenylsulfanyl)-acetic and and 301.0 mg (0.73 retool) (E)- (6R,7R)-7-amino-3-[ 1-(3-hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid trifluoroacetate in ml N,N-dimethylacetami de.

Yield: 180.0 mg (37.7%) beige powder IR(KBr): 1767,1664, 1614 cm1 MS(ISP): 654.1 (M+H÷) Example Al5 (E)-(6R,7S)-3-[1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- [2-naphthalen-2-ylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene2-carboxylic acid C02H O With 146.0 mg (0.90 retool) 1,1'-earbonyldiimidazole, 196.5 mg (0.90 retool) (naphthalen-2-ylsulfanyl)-aeetic acid and 301.0 mg (0.73 retool) (E)-(6R,7R)-7amino-3-[1-(3-hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid trifluoroacetate in 5 ml N,Ndimethylacetamide. The resulting solid was purified by column chromatoCA 02220188 1997-11-04 1,5 graphy on MCI gel (75-150 t, Mitsubishi Kasei Corporation) with a gradient of water : acetonitñle (1:0, 4:1, 3:1, 2:1, 1:1). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 65.0 mg (14.6%) beige lyophilisate IR(KBr): 1771, 1663, 1589 cm:1 MS(ISP): 602.2 (M+H÷) ETample Al6 Mixture of (E)-(6R,7R)-8-oxo-3-[(R)- and -[(S)-2-oxo-[1,3']bipyrrolidinyl-3ylidenemethyl]-8-oxo-7-[2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino]-5thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid hydrochloride CI 0 O/Z- " CO2H O Step a: Mixture of (E)-(6R,7R)-3-[(R)- and-[(S)-l'-aUyloxycarbonyl-2-oxo- [1,3']bipyrrolidinyl-3-ylidenemethyl]-8-oxo-7-[2-(2,4,5-triehloro-phenylsulfanyl)-acetyl_Amino]-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylie add ci CI H O CO2H O With 115.2 mg (0.71 Email) 1,1'-carbonyloEjmidazole, 193.4 mg (0.71 retool) (2,4,5-t ch]oro-phenylsu]£anyl)-acetic acid and 329.1 mg (0.59 retool) of a nature of (E)-(6R,7R)-3-[(R)- and -[(S)-l'-a]]y]oxycarbonyl-2-oxo- [1,3']bipTn'o]ioEn-3-ylídenemethy])-7-azmno-8-oxo-5-t a1-aza-bicyc]o[4.2.0]oct-2ene-2-carboxy]ic acid t fluoroacetate in 6 ml N,Nmethy]acetamide Prepared according to example AI.

Yield: 220.0 mg (66.0%) beige powder IR(KBr): 1774, 1678, 1624 cm"1 MS(ISP): 703.2 (M+H÷) Step b: Mixture of (E)-(6R,7R)-8-oxo-3-[(R)- and -[(S)-2-oxo-[1,3']bipyrrolidinyl3-ylidenemethyl]-8-oxo-7-[2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino]-5thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid hydrochloride CA 02220188 1997-11-04 The product prepared in step a (220.0 mg, 0.31 retool) was suspended in 12 ml dichloromethane and 124 l (0.50 retool) N,O-bis-(trimethylsilyl)-acetamide was added. After a clear solution had formed, 5.60 mg (8.56 mol) palladinm-bis-(triphenylphosphine)-dichloride, 0.36 ml (6.30 retool) acetic acid and 0.8 ml (3.0 retool) tñbutyltinhydride were added. After 45 mln 40 1 water was added to the suspension and the reaction mixture was poured under stirring on 200 ml diethyl ether, containing 2 ml of a hydrochloric acid-saturated diethyl ether solution. The solid was collected by filtration and triturated with 40 ml ethyl acetate.

Yield: 180.0 mg (87.8%) beige powder IR(KBr): 1771, 1661, 1582 cm"1 MS(ISP): 619.1 (M+H÷) ample Al7 (E)-(6R,7R)-3-[ 1-[1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1- ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-(2-phenylsulfanylacetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate F @OH ¢020 With 175.0 mg (1.08 retool) 1,1'-carbonyldiimidazole, 182.0 mg (1.08 retool) 2- (phenylthio)acetic acid and 500.0 mg (0.75 retool) (E)-(6R,7R)-7-amino-3-[1-[1- [(3-fluoro-4-hydroxy-phenylcarbamoyl)-methyl]- pyridin1-ium-4-ylmethyl]-2oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate trifluoroacetate in 4 ml N,N-dimethylacetamide. The resulting solid was puññed by column chromatography on MCI gel (75-150 Mitsubishi Kasei Corporation) with a gradient of water : acetonitñle (1:0, 4:1, 3:1, 2:1). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 90.0 mg (20.6%) beige lyophilisate IR(KBr): 1772, 1680, 1648 cm."1 MS(ISP): 704.4 (M +) Example Al8 (E)-(6R,7R)-3-[1-[1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin-1ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-[2-(4-trifluoroCA 02220188 1997-11-04 methyl-phenylsulfanyl)-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate N+.,' 0 F CO0" O With 66.8 mg (0.41 retool) 1,1,-carbonyldiimidazole, 97.3 mg (0.41 retool) 2-[4-- (trifluoromethyl)phenyltlaio]aeetic acid and 250.0 mg (0.37 retool) (E)-(6R,7R)- 7-amino-3-[ 1-[ 1-[(3-fluoro-4-hydroxy-phenylcarbamoyl)-methyl]- pyridin-1inm-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate trifluoroacetate in 4 ml N,Ndimethylacet3mide. The resulting solid was purified by column chromatography on MCI gel (75-150 t, Mitsubishi Kasei Corporation) with a gradient of water : acetonitrile (1 : 0, 4 : 1, 3 : 1, 2 : 1, 1 : 1). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 89.0 mg (30.8%) pale yellow lyophilisate IR(KBr): 1777, 1678, 1650 cm"1 MS(ISP): 772.3 (M+H +) Example Al9 (E)-(6R,7R)-7-[2-(2,5-Dichloro-phenylsulfanyl)-acetylamino]-3-[ 1- [(3-fluoro-4hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate O CI OH CO0 0 With 72.9 mg (0.45 retool) 1,1,-carbonyldiimidazole, 106.5 mg (0.45 retool) (2,5-dich]oro-phenylsulfanyl)acetìc acid and 250.0 mg (0.37 mmol) (E)- (6R,7R)-7-amino-3-[1-[1-[(3-fluoro-4-hydroxy-phenylcarbamoyl)-methyl]- pyñdin1-ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1- aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate trifluoroacetate in 4 ml N,Ndimethylacetamide. The resulting solid was purified by column chromatography on MCI gel (75-150p, Mitsubishi Kasei Corporation) with a gradient of water : acetonitrile (1 : 0, 4 : 1, 3 : 1, 2 : 1, 1 : 1). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

CA 02220188 1997-11-04 Yield: 74.5 mg (21.3%) beige lyophilisate IR(KBr): 1772, 1680, 1650 cm"1 MS(ISP): 772.3 (M+H +) ETAmple A20 (E)-(6R,7R)-3-[1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- (2-phenylsulfanyl-acety]am no)-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid Q o oe2H O With 121.1 mg (0.75 mmol) 1,1,-carbonyldiimidazole, 125.6 mg (0.75 retool) (phenyltMo)acetic add and 250.0 mg (0.62 retool) (E)-(6R,7R)-7-amino-3-[1-(3hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo- [4.2.0]oct-2-ene-2-carboxylic acid tñfluoroacetate in 4 ml N,N-dimethylacetam de. The resulting solid was purified by column chromatography on MCI gel (75-150 Mitsubishi Kasei Corporation) with a gradient of water :

acetonitrile (1 : 0, 4 : 1, 3 : 1). The organic solvent was stripped offat a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 189.5 mg (55.2%) yellow lyophilisate IR(KBr): 1764, 1664, 1612 cm-1 MS(ISP): 552.2 (M+H ÷) Example A21 (E)-(6R,7R)-7-[2-(3,4-Dimethoxy-phenylsulfanyl)-acetylamino]-3-[ 1-(3hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid CH30 ì CH30 / o o _ CO2H O With 121.1 mg (0.75 retool) 1,1,-carbonyldiimidazole, 170.6 mg (0.75 retool) 2- (3,4-dimeghoxyphenylthio)acetic acid and 250.0 mg (0.62 retool) (E)-(6R,7R)-7ami no3- [ 1- ( 3-hydroxy-benzyl)-2oxo-pyrrolidin-3 -ylidenemethyl]- 8 - oxo5-thia1-aza-bieyclo[4.2.0]oct-2-ene-2-earboxylic acid tñfluoroacetat in 4 ml N,Ndimethylacetamide. The resulting solid was puñfied by column chromatoCA 02220188 1997-11-04 graphy on MCI gel (75-150 Mitsubishi Kasei Corporation) with a gradient of water : acetonitñle (1 : 0, 4 : 1, 3 : 1). The organic solvent was stripped offat a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 175.6 mg (46.1%) pale yellow lyophilisate IR(KBr): 1766, 1664, 1588 cma MS(ISP): 612.2 (M+H +) k'br.mple A22 (E)-(6R,7R)-3-[1-[1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin-1ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-[2-(2,4,5-trichlorophenylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate CI CO2O With 60.0 mg (0.36 retool) 1,1,-carbonyldiinñdazole, 99.0 mg (0.36 retool) (2,4,5-trichloro-phenylsulfanyl)-acetic acid and 200.0 mg (0.30 mmol) (E)- (6R,7R)-7-amino-3-[1-[1-[(3-fluoro-4-hydroxy-phenylcarbamoyl)-methyl]- pyriàin-l-ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia-1aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate trifluoroacetate in 4 ml N,Ndimethylacetamide. The resulting solid was purified by column chromatography on MCI gel (75-150 Mitsubishi Kasei Corporation) with a gradient of water : acetonitril (2 : 1, 1 : 1). The organic solvent was stripped offat a rotary evaporator and OEe aqueous phase was freeze-dried.

Yield: 140.0 mg (57.8%) beige lyophilisate IR(KBr): 1763, 1666, 1645 cm-i MS(ISP): 806.3, 808.3 (M+H) ÷ The following compounds were prepared according to Example A1 E ample A23 (E)-(6R,7R)-7-[2-(Benzothiazol-2-ylsulfanyl)-acetylam no]-3-[1-(4-hydroxybenzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid imidazole salt (1:1) Yield: 66.0% beige solid MS(ISP): 609.4 (M+H)÷ IR(KBr): 1772, 1669, 1613 cm"i CA 02220188 1997-11-04 k mple A24 (E)-(6R,7R)-3-[1-(1H-Benzoimidazol-2-ylmethyl)-2-oxo-pyrrolidin-3ylidenemethyl]-8-oxo-7-(phenylsulfanylcarbonylmethyl-amino)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazol salt (1:1) Yield: 91.6% brown solid MS(ISP): 576.1 (M+H)÷ IR(KBr): 1770, 1673, 1625 cm1 mple A25 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2- (naphthalen-l-ylsulfanyl)-acetylamino]-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2ene-2-carboxylic acid imidazole salt (1:1) Yield: 41.6% beige solid MS(ISP): 536.3 (M+H)÷ IR(KBr): 1769, 1664, 1624 cm-1 example (E)-(6R,7R)-3-[1-(4-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-7-[2- (naphthalen-2-ylsulfanyl)-acetylamino]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2ene-2-carboxylic acid imidazole salt (1:1) Yield: 67.7% beige solid MS(ISP): 602.2 (M+H)÷ IR(KBr): 1771, 1667, 1614 cm"1 ple A27 (E)-(6R,7R)-7-[2-(Benzothiazol-2-ylsulfanyl)-acetylamino]-3-(1-cyclopropyl-2oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid imidazole sait (1:1) Yield: 78.5% light yellow solid MS(ISP): 543.2 (M+H)÷ IR(KBr): 1769, 1665, 1624 cm1 Example A28 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-(3,5dimethyl-phenylsulfanyl)-acetylamino]-8-oxo-5-thía1-aza-bicyclo[4.2.0]oct-2ene-2-carboxylic acid imidazole salt (1:1) Yield: 67.0% beige solid MS(ISP): 514.3 (M+H)÷ IR(KBr): 1765, 1653, 1621 cm-1 CA 02220188 1997-11-04 v., -mple (E)-(6R,7R)-7-[2-(4-Bromo-phenylsulfanyl)-acetylamino]-3-(1-cyclopropyl-2oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid imidazole salt (1:1) Yield: 75.5% light yellow solid MS(ISP): 566.1 (M+H)÷ IR(KBr): 1767, 1661, 1622 cm"1 F«-mpleA30 (E)-(6R,7R)-7-[2-(Benzooxazol-2-ylsulfanyl)-acetylamino]-3-(1-cyclopropyl-2oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid imidazole salt (1:1) Yield: 67.2% beige solid MS(ISP): 527.1 (M+H)+ -1 IR(KBr): 1770, 1670, 1625 cm E ample A31 (E)-(6R,7R)-7-[2-(4-Bromo-phenylsulfanyl)-acetylamino]-8-oxo-3-[2-oxo-1- (2,2,2-trifluoro-ethyl)-pyrrolidin-3-ylidenemethyl]-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) Yield: 86.3% yellow powder MS(ISP): 608.3 (M+H)÷ IR(KBr): 1769, 1682, 1611 cm"1 E mple A32 (E)-(6R,7R)-8-Oxo-3-[2-oxo-1-(2,2,2-trifluoroethyl)-pyrrolidin-3ylidenemethyl]-7-(2-phenylsulfanyl-acetylamino)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) Yield: 78.6% yellow powder MS(ISP): 528.3 (M+H)÷ IR(KBr): 1771, 1681, 1612 cm1 -,-ple A33 (E)-(6R,7R)-3-(1-Benzyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7-(2phenylsulfanyl-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) Yield: 85.0% brown solid MS(ISP): 536.4 (M+H)÷ CA 02220188 1997-11-04 IR(KBr): 1767, 1667, 1622 cm"1 F. .mple (E)-(6R,7R)-3-[1-(4-Fluoro-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- (2-phenylsulfanyl-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid im dazole salt (1:1) Yield: 91.0% beige solid MS(ISP): 554.5 (M+H)÷ IR(KBr): 1766, 1665, 1622 cm"1 k -mpleA (E)-(6R,7R)-3-[1-(4-Methoxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- (2-phenylsulfanyl-acetylamino)-5-thia-1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid imidazole salt (1:1) Yield: 87.1% beige solid MS(ISP): 566.5 (M+H)÷ IR(KBr): 1773, 1672, 1611 cm1 EY-mple A36a (E)-(6R,7R)-3-[1-(4-Allyloxycarbonylam no-benzyl)-2-oxo-pyrrolidin-3ylidenemethyl]-8-oxo-7-(2-phenylsulfanyl-acetylamino)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) RO-65-2011 Yield: 82.3% brown solid MS(ISP): 652.5 (M+NH4 )÷ IR(KBr): 1770, 1726, 1667, 1613 cm1 pleA? b (E)-(6R,7R)-3-[1-(4-Am no-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- (2-phenylsulfanyl-acetylamino)-5-thia-1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid (E)-(6R,7R)-3-[ 1-(4-allyloxycarbonylamino-benzyl)-2-oxo-pyrrolidin-3ylidenemethyl]-8-oxo-7-(2-phenylsulfanyl-acetylamino)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) (400.0 mg, 0.57 retool) was suspended in 20 ml dichloromethane and treated with N,Obis(trimethylsilyl)-tñfluoroacetamide (240.7 ml, 9.11 mmol). After 15 relu, bis(triphenylphosphine)palladium(II) chloride (10.0 mg, 0.014 mmol), acetic acid (0.65 ml, 11.4 retool) and tributyltin hydride (1.53 ml, 5.70 mmol) were added. After 45 rein the suspension was poured under stirring on 250 ml diethyl ether, containing 3 ml of a hydrochloric acid-saturated diethyl ether CA 02220188 1997-11-04 solution and was stirred for th. The solid material was collected by filtration, suspended in 4 ml water : acetonitrile (1 : 1) and the pli was adjusted to 2 with 1M hydrochloric acid. To thi suspension an equal mount of MCI gel (75-150 t, Mitsubishi Kasei Corporation) was added, the organic solvent was evaporated and the residue was chromatographed on MCI gel (75-150 Mitsubishi Kasei Corporation) with a gradient of water : acetonitrile (9 : 1, 4 :

1, 2 : 1, 1 : 1, 1 : 3). Thi organic solvent was evaporated and the aqueous phase was freeze-dried.

Yield: 18.0% beige lyophilisate MS(ISP): 551.5 (M+H)÷ IR(KBr): 1769, 1667, 1626 cm"1 E mpleA (E)-(6R,7R)-3-[1-(1,1-Dioxo-tetrahydro-thiophen-3-yl)-2-oxo-pyrrolidin-3ylidenemethyl]-8-oxo-7-(2-phenylsulfanyl-acetylamino)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole sait (1:1) (1:1 mixture of epimers) Yield: 56.1% beige solid MS(ISP): 581.4 (M+H)÷ IR(KBr): 1771, 1673, 1618 cma E mple A2,Sa (E)-(6R,7R)-3-[1-(1-Allyloxycarbonyl-azetidin-3-yl)-2-oxo-pyrrolidin-3ylidenemethyl]-8-oxo-7-(2-phenylsulfanyl-acetylamino)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) Yield: 74.5% beige lyophilisate MS(ISP): 585.5 (M+H)÷ -1 IR(KBr): 1775, 1678, 1626 cm exemple A,38b (E)-6R,7R)-3-(1-Azetidin1-illm-3-yl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7- (2-phenylsulfanyl-acetylamino)-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate (E)-(6R,7R)-3-[1-(1-allyloxycarbonyl-azetidin-3-yl)-2-oxo-pyrrolidin-3ylidenemethyl]-8-oxo-7-(2-phenylsulfanyl-acetylamino)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) (448.0 mg, 0.69 retool) was dissolved in 20 ml dichloromethane. Acetic acid (0.79 ml, 3.70 retool), bis(triphenylphosphine)palladium(II) chloride (11.9 mg, 0.017 rnmo1) and tñbutyltin hydride (1.86 ml, 6.90 retool) were added successively.

CA 02220188 1997-11-04 Ær 45 rein the suspension was poured under stirring on 250 ml diethyl ether, containing 3 mi of a hydrochloric acid-saturated diethyl ether solution and was stirred for lh. The solid material was collected by filtration, suspended in water : acetonitñle (1 : 1) and the pli was adjusted to 2 with 1N hydrochloric acid. To the suspension an equal amount of MCI gel (75-150g, Mitsubishi Kasei Corporation) was added, the organic solvent was evaporated and the residue was chromatographed on MCI gel (75-150g, Mitsubishi Kasei Corporation) with a gradient of water : acetonitrile (9 : 1, 4 :

1, 2 : 1, 1 : 1, 1 : 3). The organic solvent was evaporated and the aqueous phase was freeze-dried.

Yield: 23.2% beige lyophilisate MS(ISP): 501.4 (M+H)+ IR(KBr): 1766, 1673, 1620 cm:1 ample A39 (E)-(6R,7R)-3-[1-(4-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- (2-phenylsulfanyl-acetylam no)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid imidazole sait (1:1) Yield: 68.0% yellow solid MS(ISP): 552.5 (M+H)÷ IR(KBr): 1773, 1667, 1614 cm"1 E ample A40 (E)-(6R,7R)-8-Oxo-3-(2-oxo1-phenylcarb moylmethyl-pyrrolidin-3ylidenemethyl)-7-(2-phenylsulfanyl-acetylamino)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole sait (1:1) Yield: 52.0% brown solid MS(ISP): 579.4 (M+H)÷ IR(KBr): 1770, 1665, 1599 cm"1 Æxample A41a (E)-(6R,7R)-3-[(R)- l'-Allyloxycarbonyl-2-oxo-[ 1,3']bipyrrolidinyl-3ylidenemethyl]-8-oxo-7-(2-phenylsulfanyl-acetylamino)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) Yield: 81.3% brown solid MS(ISP): 562.3 (M+H)÷ -1 IR(KBr): 1776, 1670, 1631 cm CA 02220188 1997-11-04 E'xample A41b (E)-(6R,7R)-8-Oxo-3-[(R)-2-oxo-[ 1,3']bipyrrolidinyl-3-ylidenemethyl]-7-(2phenylsulfanyl-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylicacid hydrochloride (1:1) (E)-(6R,7R)-3-[(R)- r-allyloxycarbonyl-2-oxo-[1,3']bipyrrolidinyl-3ylidenemethyl]-8-oxo-7-(2-phenylsulfanyl-acetylamino)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (370.0 mg, 0.62 retool) was dissolved in 20 ml dichloromethane and treated successively with bis(triphenylphosphine)palladium(II) chloride (10.9 mg, 0.015 rnmol), acetic acid (0.71 ml, 12.4 retool) and tñbutyltin hydñde (1.67 ml, 6.20 mmol). After m u, the suspension was poured on 250 ml diethyl ether containing 3 ml of a hydrochloric acid-saturated diethyl ether solution and stirred for lh. The suspension was filtered, the solid material was triturated with ethyl acetate for lh and dried in high vacuum.

Yield: 25.5% beige solid MS(ISP): 515.3 (M+H)+ IRçKBr): 1776, 1666, 1632 cma k mple A42 (E)-(6R,7R)-7-[2-(4-Chloro-phenylsulfanyl)-acetylamino]-3-(1-cyclopropyl-2oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thial-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid imldazole salt (1:1) Yield: 70.7% light yellow solid MS(ISP): 520.4 (M+H)+ IR(KBr): 1775, 1667, 1625 cm1 The following compounds were prepared according to Example Al0 F mple A43 (E)-(6R,7R)-8-Oxo-3-[2-oxo1-(2-oxo-oxazolidin-3-yl)-pyrrolidin-3ylidenemethyl]-7-(2-phenylstùfanyl-acetylamino)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole sait (1:1) Yield: 36.0% beige lyophilisate MS(ISP): 548.4 (M+H)÷ IEçKBr): 1770, 1689, 1612 cm"1 Example A44 (E)-(6R,7R)-8-Oxo-3-(2-oxo1-pyridin-2-yl-pyrrolidin-3-ylidenemethyl)-7-(2phenylsulfanyl-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) CA 02220188 1997-11-04 Yield: 18.0% brown lyophilisate MS(ISP): 523.5 (M+H)÷ IR(KBr): 1770, 1680, 1610 cm"1 ExampleA45 (E)-(6R,7R)-3-[1-(6-Methoxy-pyridin-3-yl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8oxo-7-(2-phenylsulfanyl-acetylamino)-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid imidazole salt (1:1) Yield: 12.0% beige lyophilisate MS(ISP): 553.5 (M+H)+ IR(KBr): 1769, 1677, 1619 cm-1 E ample A46 (E)-(6R,7R)-7-[2-(1H-Benzoimidazol-2-ylsulfanyl)-acetylamino]-3-[1-(4hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) Yield: 11.2% beige lyophilisate MS(ISP): 592.5 (M+H)÷ IR(KBr): 1769, 1664, 1614 cm-1 Example A47 (E)-(6R,7R)-3-(1-Isobutyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7-(2phenylsulfanyl-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) Yield: 15.8% colorless lyophilisate MS(ISP): 502.4 (M+H)÷ -1 IR(KBr): 1768, 1660, 1625 cm simple A48 (E)-(6R,7R)-3-(1-Cyclohexylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7- (2-phenylsulfanyl-acetylami no)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid imidazole salt (1:1) Yield: 10.0% colorless lyophilisate MS(ISP): 542.4 (M+H)÷ k-sample A49 (E)-(6R,7R)-3-[1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- (3-phenyl-propionylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid CA 02220188 1997-11-04 Yield: 17.0% beige lyophilisate MS(ISP): 534.4 (M+H)÷ IR(KBr): 1778, 1662, 1602 cm"1 k'bxample A50 (E)-(6R,7R)-3-[(3-Hydroxy-benzyl)-oxo-pyrrolidin-3-ylidenemethyl]-7-[2-(4hydroxymethyl-phenoxy)-acetylamino]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2ene-2-carboxylic acid imidazole salt (1:1) Yield: 21.2% beige lyophilisate MS(ISP): 566.4 (M+H)÷ IR(KBr): 1773, 1665, 1588 cma Example A51 (E)-(6R,7R)-3-[1-[1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1- ium-4-ylmethyl}]-2-oxo-pyrrolidin-3-ylidenemethyl]- 7-[2-(4-hydroxymethylphenoxy)-acetylamino]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate Yield: 14.1% beige lyophilisate MS(ISP): 718.3 (M+H)÷ IR(KBr): 1769, 1681, 1613 cm"1 mple A52 (E)-(6R,7R)-7-(2-Benzoylamino-acetylamino)-3-[1-(3-hydroxy-benzyl)-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid imidazole salt (1:1) Yield: 32% beige lyophilisate MS(ISP): 562.0 (M+H)+ IR(KBr): 1771, 1658, 1602 cm1 Example A53 (E)-(6R,7R)-3-[1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- (2-phenylam no-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) Yield: 20.5% beige lyophilisate MS(ISP): 535.4 (M+H)÷ IR(KBr): 1780, 1670, 1608 cm1 CA 02220188 1997-11-04 mple A54 (E)-(6R,7R)-3-[1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- (2-phenoxy-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:0.8) Yield: 34.6% beige lyophilisate MS(ISP): 536.2 (M+H)÷ IR(KBr): 1776, 1673, 1600 cma E mpleA55 (6R,7R)-3-[(E)- 1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7- [(Z)-2-styrylsulfanyl-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid Yield: 31.6% beige lyophilisate MS(ISP): 578.4 (M+H)÷ IR(KBr): 1775, 1663, 1619 cm=1 ample A56 (6R,7R)-3-[(E)- 1-[ 1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1- inm-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-[(Z)-2styrylsulfanyl-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate Yield: 14.2% beige lyophilisate MS(ISP): 730.5 (M+H)÷ IR(KBr): 1769, 1677, 1642 cm-1 F« mple A57 (E)-(6R,7R)-7-[2-(4-Chloro-phenylsulfanyl)-acetylamino]-3-[ 1-(3-hydroxybenzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid imidazole salt (1:1) Yield: 29.0% light yellow lyophilisate MS(ISP): 586.3 (M+H)÷ IR(KBr): 1769, 1669, 1600 cm1 ample A58 (E)-(6R,7R)-7-[2-(4-Bromo-phenylsulfanyl)-acetylamino]-3-[1-[1-[(3-fluoro-4hydroxy-phenylcarbamoyl)-methyl]-pyridin-l-ium-4-ylmethyl]-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate Yield: 13.3% beige lyophilisate MS(ISP): 748.1 (M+H)÷ CA 02220188 1997-11-04 IR(KBr): 1769, 1676, 1642 cm"1 F.T mple A59 (E)-(6R,7R)-7-[2-(3,5-Dimethyl-phenylsulfanyl)-acetylamino]-3-[1-[1-[(3-fluoro4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1-inm-4-ylmethyl]-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate Yield: 24.0% light brown lyophilisate MS(ISP): 732.4 (M+H)÷ IR(KBr): 1766, 1667, 1644 cm"1 Example A60 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7-[2- (pyridin-4-ylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid imidazole salt (1:1) Yield: 46.0% light yellow lyophilisate MS(ISP): 486.4 (M+H)÷ IR(KBr): 1769, 1664, 1619 cm1 E Ample A61 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7-[2- (pyñdin-4-ylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid imidazole sait (1:1) Yield: 41.0% light yellow lyophilisate MS(ISP): 487.2 (M+H)÷ IR(KBr): 1772, 1672, 1623 cm"1 pleA62 (E)-(6R,7R)-7-[2-(4-Fluoro-phenylsulfanyl)-acetylamìno]-8-oxo-3-(2-oxo1- phenylcarbamoylmethyl-pyrrolidin-3-ylidenemethyl)-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) (4-Fluorophenylthio)acetic acid (102.7 mg, 0.55 retool) was dissolved in N,Ndimethylacetamide and 1,1'-carbonyldiimidazole (89.0 mg, 0.55 retool) was added in a single portion. The solution was stirred for 20 rein, then (E)- (6R,7R)-7-amino-8-oxo-3-(2-oxo-l-phenylcarbamoylmethyl-pyrrolidin-3ylidenemethyl)-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid was added. After 6h the mixture was poured on 250 ml diethyl ether and the solid mateñal was collected by filtration. The solid was converted into its sodium sait by suspending it in 6 ml water and adjusting the pli te 7 with 1M sodium CA 02220188 1997-11-04 hydroxide solution. The solution was chromatographed on MCI gel (75-150g, Mitsubishi Kasei Corporation) with a gradient of water : acetonitrile (10 : 1, 8 : 2, 7 : 3). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 75.2% yellow lyophilisate MS(ISP): 597.2 (M+H)+ IR(KBr): 1770, 1668, 1630 cm"1 The following compounds were prepared according to Example A62 Example A{ì3 (E)-(6R,7R)-8-Oxo-3-(2-oxo1-phenyl-pyrrolidin-3-ylidenemethyl)- 7-(2phenylsulfanyl-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 56.3% beige lyophilisate MS(ISP): 522.2 (M+H)+ IR(KBr): 1765, 1682, 1622 cm"1 YampleA64 (E)-(6R,7R)-3-[1-(2-Methyl-allyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-7-(2phenylsulfanyl-acetylamino)-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 40.8% beige lyophilisate MS(ISP): 500.2 (M+H)÷ IR(KBr): 1765, 1667, 1614 cm"1 A65 (E)-(6R,7R)-7-[2-(Benzooxazol-2-ylsulfanyl)-acetylamino]-3-[1-(4-hydroxybenzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid sodium salt (1:1) Yield: 43.3% beige lyophilisate MS(ISP): 593.2 (M+H)÷ IR(KBr): 1770, 1670, 1615 cm"1 -e-ple A66 (E)-(6R,7R)-7-(2-Benzylsulfanyl-acetylamino)-3-[ 1-(4-hydroxy-benzyl)-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0] oct-2-ene-2carboxylic acid sodium salt (1:1) Yield: 38.7% beige lyophilisate MS(ISP): 583.3 (M+H)÷ CA 02220188 1997-11-04 IR(KBr): 1764, 1664, 1614 cm1 A67 (E)-(6R,7R)-7-[2-(5-Acetylamíno-[1,3,4]thiaiazol-2-ylsulfanyl)-acetylamino]- 3-[ 1-(4-hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 40.4% yellow lyophilisate MS(ISP): 617.2 (M+H)÷ IR(KBr): 1766, 1657, 1614 cm"1 E- ,-ple ASS (E)-(6R,7R)-3-[1-(4-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-7-(2methylsulfanyl-acetylamino)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid sodium salt (1:1) Yield: 43.0% beige lyophilisate MS(ISP): 490.3 (M+H)+ IR(KBr): 1764, 1664, 1614 cm-1 E--mple A69 (E)-(6R,7R)-7-[2-(4-Bromo-phenylsulfanyl)-acetylamino]-8-oxo-3-(2-oxo1- phenyl-pyrrolidin-3-ylidenemethyl)-5-thia-1-aza-bicyclo [4.2.0]oct-2-ene-2carboxylic acid sodium salt Yield: 53.0% yellow lyophilisate MS(ISP): 602.1 (M+H)÷ IR(KBr): 1764, 1673, 1618 cm-1 Example AT0 (E)-(6R,7R)-7-[2-(Naphthalen-2-ylsulfanyl)-acetylamino]-8-oxo-3-[2-oxo1- (2,2,2-trifluoro-ethyl)-pyrrolidin-3-ylidenemethyl]-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1); compound with imidazole (1:0.5) Yield: 28.3% yellow lyophilisate MS(ISP): 578.1 (M+H)÷ IRCKBr): 1770, 1682, 1620 cm"1 The following compound was prepared according to l -mple AI E ample AT1 1:1 Mixture of (E)-(6R,7R)-8-0xo-3-[2-oxo-l-[(R)- and-[(S)-tetrahydro-furan-2CA 02220188 1997-11-04 ylmethyl]-pyrrolidin-3-ylidenemethyl]-7-(2-phenylsulfanyl-acetylam no)-5thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid imidazole salt (1:1) Yield: 88.0% yellow lyophilisate MS(ISP): 530.2 (M+H)÷ IR(KBr): 1772, 1672, 1619 cm"1 MethodB kmmpleB1 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7- ( 2-thiophen2-ylacetylamino)-5 -thia1 - aza-bicyclo [4.2.0] oct2 -ene2- carb oxylic acid H COOH 0 To a suspension of 300.0 mg (0.86 retool) (E)-(6R,7R)-7-am no-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-azabicyclo- [4.2.0]oct-2-ene-2-carboxylic acid in 8 ml N,N-dimethylformamide was added 80.0 mg (0.95 retool) sodium bicarbonate. After cooling to 0°C 101.0 1 (0.95 retool) 2-thiopheneacetyl chloride were added. After 1.5 h the solvent was stripped off at a rotary evaporator and the residue was poured on diethyl ether. The solid was collected by filtration and was triturated with 25 ml ethyl acetate and 15 ml water for 1.5 h.

Yield: 150.0 mg (36.9%) beige powder IR(KBr): 1779, 1669, 1626, 1540 cm-1 MS(ISP): 474.2 (M+H÷) Example 132 Mixture of (E)-(6R,7R)-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[(R)- and [(S)-2,3-diphenyl-propionylamino)]-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid H C02H 0 From 100.0 mg (0.29 retool) (E)-(6R,7R)-7-amino-3-(1-cyclopropylmethyl-2oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-azabicyclo[4.2.0] oct-2-ene-2CA 02220188 1997-11-04 carboxylic acid, 26.4 mg (0.31 retool) sodium bicarbonate and 77.0 mg (0.31 retool) 2,3-diphenylpropionyl chloride in 8 ml N,N-dimethylformamide.

Yield: 82.0 mg (51.4%) yellow powder IR(KBr): 1782, 1671 cm"1 MS(ISP): 558.4 (M+H÷) mpleB3 (E)-(6R,7R)-7-[2-(4-Chloro-phenylsulfanyl)-2-methyl-propionylamlno]-3-(1cyclopropylmethyl-2oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Prepared according to example B1.

The beige solid was suspended in water and the pli was adjusted to 7 with 1M sodium hydroxide solution. The resulting solution was chromatographed on MCI gel (75-150 t, Mitsubishi Kasei Corporation) with a gradient of water : acetonitrile (9 : 1, 8 : 2, 7 : 3). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 56.0% beige lyophilisate IR(KBr): 1765, 1669, 1620 cm"1 MS(ISP): 562.3 (M+H÷) Method C RTample Cla (E)-(6R,7R)-7-(2-Bromo-acetylamino)-3-(1-[1-[(3-fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate O N +f' /F Br""N NH Çí \ N H- " r õ -" "'") / OH o I \ cooo To a suspension of 280.0 mg (0.42 retool) (E)-(6R,7R)-7-amino-3-[1-[1-[(3tluoro-4-hydroxy-phenylcarbamoyl)-methyl]- pyridin1-ium-4-ylmethyl]-2oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2earboxylate tritluoroaeetate in 4 ml diehloromethane were added 0.37 gl (1.40 130 retool) N,O-bis-(trimethylsilyl)-tritluoroaeetamide. Æer a clear solution had formed, 36.8 gl (0.42 retool) bromoacetyl bromide were added and the reaction mixture was stirred for 3 h. To this solution 25.6 1 (1.42 retool) water and 25 ml diethyl ether was added. The precipitate was filtered off and washed with diethyl ether.

CA 02220188 1997-11-04 Yield: 210.0 mg (77.8%) beige powder IR(KBr): 1782, 1686, 1643 cm1 MS(ISP): 676.2 (M+H÷) simple Clb (E)-(6R,7R)-3-[1-[1-[(3-Fluoro-4-hydroxy-phenylcarb_amoyl)-methyl]-pyridin1- inm-4-ylmethyl]-2-oxo-pyrrolidìn-3-ylidenemethyl]-7-[2-(4-methyl-4H-[ 1,2,4]- tr azo -3-y su fany )-acety amino]-8-oxo-5-thia- -aza-bicyc o[4.2. ]oct-2-ene-2carboxylate N,,í-... N/ / 0 F "N "S O'Õ NHNH oH COO - O To a solution of 230.0 mg (0.34 mmol) (E)-(6R,7R)-7-(2-Bromo-acetylarnino)-3- ( 1-[ 1- [(3-fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylate in 5 ml N,N-dimethylformamide was added 40.0 mg (0.37 retool) 4-methyl-4H-[1,2,4]triazole-3-thiol. After 1 h 47.0 gl (0.34 retool) tñethyl amine was added and stirring was continued for 12 h. The solution was poured on diethyl ether and the resulting solid was purified by column chromatography on MCI gel (75-150g, Mitsubishi Kasei Corporation) with a gradient of water : acetonitríle (1 : 0, 4 : 1, 3 : 1, 2 : 1). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 48.4 mg (20.0%) beige lyophilisate -1 IR(KBr): 1780, 1660, 1647 cm MS(ISP): 709.3 (M+H÷) Example 132 (E)-(6R,7R)-3-[1-[ 1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl]-pyridin1- ium-4-ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl]-7- [2-(4-hydroxyphenylsulfanyl)-acetylanñno]-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate (E)-(6R,7R)-7-(2-bromo-acetylanñno)-3-[ 1-[1-[(3-fluoro-4-hydroxyphenylcarbamoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2oxo-pyrrolidin-3ylidenemethyl]-8-oxo-5-thia1-aha-bicyclo[4.2.0]oct-2-ene-2-carboxylate (see ex. Cla) (300.0 mg, 0.45 retool) was dissolved in 3 ml N,Ndimethylformamide, then 4-mercaptophenol (58.0 mg, 0.45 mmol), followed by triethylamine (62.7 ml, 0.45 mmol) was added. After 22h the reaction CA 02220188 1997-11-04 mixture was poured on diethyl ether and the solid was collected by filtration.

The brown solid mateñal was suspended in water : acetonitrile (1 : 1) and chromatographed on MCI gel (75-1501z, Mitsubishi Kasei Corporation) with a gradient of water : acetonitrile (9 : 1, 4 : 1, 3 : 1). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 14.0% beige solid MS(ISP): 720.4 (M+H)÷ IR(KBr): 1769, 1671, 1643 cm"1 exemple C3 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7- [2-(thiophen-2-ylsulfanyl)-acetylamino]-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid Prepared according to example C1 Yield: 24.2% beige lyophilisate MS(ISP): 506.2 (M+H)÷ IR(KBr): 1781, 1719, 1667 cmd l .,-ple C4a (E)-(6R,7R)-7-(2-Bromo-acetylamino)-3-(1-cyclopropyl-2-oxo-pyrrolidin-3ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) (E)-(6R,7R)-7-amino-3-[(1-cyclopropyl-2-oxo-3-pyrrolidinylidene)methyl]-8oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid tñfluoroacetate (1 :

0.25) (8.0 g, 22.0 retool) was suspended in 100 ml dichloromethane and Nmethyl-N-(trimethylsilyl)trifluoroacetamide (9.0 ml, 48.4 mmol) was added.

After 45 rein a solution had formed which was cooled to 0°C and treated with bromoacetyl bromide (2.10 ml, 24.2 mmol). After 30 rein the ice-bath was removed and the reaction was stirred at ambient temperature for 2.5h. The volatile components were evaporated and the residue was converted into its sodium salt by suspending it in water and adjusting the pli to 6.5 with 1M sodium hydroxide solution. The solution was freeze-dried and the crude lyophilisate was purified by reversed phase chromatography (RP-18 LiChroPrep gel) with a gradient of water : acetonitril (10 : 0, 9 : 1). The organic solvent was evaporated and the aqueous phase was freeze-dried.

Yield: 60.0% light yellow lyophilisate MS(ISP): 465.2 (M+H)+ IR(KBr): 1766, 1672, 1620 cIn-1 CA 02220188 1997-11-04 Example CAb (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-(1Himidazol-2-ylsulfanyl)-acetylamino]-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene2-carboxylic acid sodium salt (1:1) (E)-(6R,7R)-7-(2-bromo-acetylamino)-3-(1-cyclopropyl-2-oxo-pyrrolidin-3ylidenemethyl)-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) (200.0 mg, 0.42 retool) was dissolved in 4 ml N,Ndimethylformamide and 2-mercaptoimidazole sodium salt (56.2 mg, 0.46 mmol) was added in a single portion. After 15h the solvent was stripped off at a rotary evaporator. The residue was dissolved in water and purified by reversed phase chromatography (RP-18 LiChroPrep gel) with a gradient of water : acetonitril (10 : 0, 9 : 1, 8 : 2). The organic solvent was evaporated and the aqueous phase was freeze-dried.

Yield: 82.2% colorless lyophilisate MS(ISP): 476.1 (M+H)÷ IR(KBr): 1764, 1664, 1620 cml The following compounds were prepared according to Example C4 E mple C5 (E)-(6R,7R)-7-[2-(4-Carboxy-phenylsulfanyl)-acetylamino]-3-(1-cyclopropyl-2oxo-pyrrolidin-3-ylìdenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid sooEnm salt (1:2) Yield: 57.6% yellowish lyophilisate MS(ISP): 530.2 (M+H)+ IRçKBr): 1755, 1658, 1589 cm"1 mple C6 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-(4hydroxy-phenylsulfanyl)-acetyl mluo]-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2ene-2-carboxylic acid sodium salt (1:1) Yield: 72.2% yellow lyophilisate MS(ISP): 502.0 (M+H)÷ -1 IR(KBr): 1764, 1661, 1620 cm ample C7 (E)-(6R,7R)-7-[2-(1-Carboxymethyl1H-tetrazol-5-ylsulfanyl)-acetylamino]-3- (1-cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid disodium sait Yield: 49.1% beige lyophilisate CA 02220188 1997-11-04 MS(ISP): 558.2 (M+H)+ -1 IR(KBr): 1764, 1622 cm (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7-[2-(1phenyl1H-tetrazol-5-ylsulfanyl)-acetylam no]-5-thia1-aza-bicyclo[4.2.0] oct-2ene-2-carboxylic acid sodi!lm salt (1:1) Yield: 81.8% colorless lyophilisate MS(ISP): 554.2 (M+H)+ IR(KBr): 1764, 1672, 1619 cm"1 C9 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7-[2-(2oxo-3,7-dihydro-2H-purin-6-ylsulfanyl)-acetylamino]-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 40.6% light beige lyophilisate MS(ISP): 544.4 (M+H)+ IR(KBr): 1762, 1625 cm-1 Example C10 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7-[2- (pyrimidin-2-ylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid sodi,,m salt (1:1) Yield: 27.0% colorless lyophilisate MS(ISP): 488.4 (M+H)÷ -1 IR(KBr): 1763, 1671, 1618 cm F«ample Cll (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-(1-methyl1H-tetrazol-5-ylsulfanyl)-acetylamino]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2ene-2-carboxylic acid sodium sait (1:1) Yield: 38.1% colorless lyophilisate MS(ISP): 492.4 (M+H)+ IR(KBr): 1764, 1678, 1618 cm"1 Emple C12 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-(4hydroxy-pyrimidin-2-ylsulfanyl)-acetylamino]-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) CA 02220188 1997-11-04 Yield: 47.3% light yellow lyophílisate MS(ISP): 504.4 (M+H)÷ IR(KBr): 1762, 1670, 1616 cm1 ETAmple C13 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-[2-(3,4dihydroxy-phenyl)-5-methyl-[ 1,2,4] triazolo[ 1,5-a]pyrimidin-7-ylsulfanyl]- acetylam no]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 27.3% colorless lyophilisate MS(ISP): 650.4 (M+H)÷ IR(KBr): 1768, 1660, 1618 cm"1 Example C14 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-(4-methyl5-trifluoromethyl-4H-[1,2,4]triazol-3-ylsulfanyl)-acetylamino]-8-oxo-5-thia1- aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium sait (1:1) Yield: 48.6% light yellow lyophilisate MS(ISP): 559.4 (M+H)+ IR(KBr): 1763, 1672, 1618 cm1 F.Tample C15a (E)-(6R,7R)-7-(2-Bromo-acetylamino)-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin3-ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium sait (E)-(6R,7R)-7-am no-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3ylidenemethyl)-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (3.50 g, 10.0 retool) was suspended in 50 ml dichloromethane and Nmethyl-N-trimethylsilyltrifluoroacetamide (4.62 ml, 25.0 retool) was added dropwise. Airer lh a solution had formed which was cooled to 0°C. To this solution bromoacetyl bromide (1.04 ml, 12.0 mmol) was added. Atìer 30 rein the ice-bath was removed and the solution was stirred for 4h at room temperature. After evaporation of the volatile components, the residue was suspended in water and the pli was adjusted to 6.8 with 1N sodium hydroxide solution and freeze-dried. The crude lyophilisate was purified by reversed phase chromatography (RP-18 LiChroPrep gel) with a gradient of water : acetonitñle (9 : 1, 8:2, 7 : 3). The organic solvent was evaporated and the aqueous phase was freeze-dried.

Yield: 67.1% yellow lyophilisate CA 02220188 1997-11-04 MS(ISP): 470.0 (M+H)÷ IR(KBr): 1766, 1665, 1622 cm"1 The following compounds were prepared according to Example C4 E mmple C15b (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolídin-3-ylidenemethyl)-8-oxo-7- [2-(3H-[1,2,3]tñazol-4-ylsulfanyl)-acety] amino]-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid sodium salt (1:1) Yield: 52.1% yellowish lyophilisate MS(ISP): 491.4 (M+H)÷ IR(KBr): 1764, 1664, 1619 cm"1 ample C16 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-[2- (3,4-dihydroxy-phenyl)-5-methyl-[1,2,4]triazolo[ 1,5-a]pyrimidin-7-ylsulfanyl]- acetylam uo]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 38.3% colorless lyophilisate MS(ISP): 664.1 (M+H)÷ IR(KBr): 1766, 1665, 1593 cm"1 F«Ample C17 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7- [2-(1H-[1,2,4]triazol-3-ylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid sodium salt (1:1) Yield: 51.2% yellowish lyophilisate MS(ISP): 491.4 (M+H)÷ IR(KBr): 1765, 1664, 1619 cm=1 Example C18 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7- [2-(1H-pyrazolo[3,4-d]pyrimidin-4-ylsulfanyl)-acetylamino]-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 20.1% yellowish lyophilisate MS(ISP): 542.3 (M+H)÷ IR(KBr): 1764, 1666, 1625 cm"1 Example C19 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-(4CA 02220188 1997-11-04 methyl-5-tñfluoromethyl-4H-[ 1,2,4]triazol-3-ylsulfanyl)-acetylamino]-8-oxo-5thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 52.2% yellowish lyophilisate MS(ISP): 573.3 (M+H)÷ IR(KBr): 1763, 1665, 1612 cm-1 ample C20 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-(1methyl1H-tetrazol-5-ylsulfanyl)-acetylamino]-8-oxo-5-thia1-aza bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 50.8% yellowish lyophilisate MS(ISP): 506.3 (M+H)÷ IR(KBr): 1763, 1667, 1615 cm:1 F«ample C21a Mixture of (E)-(6R,7R)-7-[(R)- and -[(S)-2-bromo-propionylamino]-3-(1cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1), see ex. Dla ample C21b Mixture of (E)-(6R,7R)-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3ylidenemethyl)-7-[(R)- and-[(S)-2-(4-methyl-5-trifluoromethyl-4H- [ 1,2,4]triazol-3-ylsulfanyl)-propionylamino]-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Prepared according to example C4 Yield: 55.0% colorless lyophilisate MS(ISP): 587.3 (M+H)÷ IR(KBr): 1765, 1670, 1618 cm'1 ample C22 Mixture of (E)-(6R,7R)-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3ylidenemethyl)-8-oxo-7-[(R)- and -[(S)-2-(3H-[1,2,3]triazol-4-ylsulfanyl)- propionylamino]-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 63.3% beige lyophilisate MS(ISP): 505.2 (M+H)÷ IR(KBr): 1764, 1657, 1621 cm"1 CA 02220188 1997-11-04 ETample C23 (E)-(6R,7R)-7-[2-(1H-Benzoimidazol-2-ylsulfanyl)-acetylamino]-3-(1cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodi11 m salt (1:1) (E)-(6R,7R)-7-(2-bromo-acetylamino)-3-(1-cyclopropyl-2-oxo-pyrrolidin-3ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium sait (1:1) (see ex. C4a) (200 mg, 0.42 retool) was dissolved in 4 ml N,N-dimethylformamide and 2-mercaptobenzimidazole sodium salt (79.2 mg, 0.46 retool) was added in a single portion. Æer completion of the reaction, the solvent was stripped off at a rotary evaporator. The residue was dissolved in water and chromatographed on MCI gel (75-150 Mitsubishi Kasei Corporation) with a gradient of water : acetonitril (10 : 1, 9 : 1). The organic solvent was evaporated and the aqueous phase was freeze-dried.

Yield: 57.0% beige lyophilisate MS(ISP): 526.0 (M+H)÷ IR(KBr): 1763, 1668, 1617 cm-1 The following compounds were prepared according to Example C23 F rample C24 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7-[2-(1Hpyrazolo [3,4-d]pyrimidin-4-ylsulfanyl)-acetylamino]-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 20.0% light yellow lyophilisate MS(ISP): 528.4 (M+H)+ IR(KBr): 1764, 1667, 1618 cm1 ample C25 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-(2,6dimethyl-5-oxo-2,5-dihydro-[1,2,4]triazin-3-ylsulfanyl)-acetylamino]-8-oxo-5thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 33.3% beige lyophilisate MS(ISP): 533.4 (M+H)+ IR(KBr): 1762, 1629, 1478 cm-1 Example C26 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-(2hydroxymethyl-5-methyl-[1,2,4]triazolo[ 1,5-a]pyrimìdin-7-ylsulfanyl)- acetylamino]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) CA 02220188 1997-11-04 Yield: 45.5% colorless lyophilisate MS(ISP): 572.5 (M+H)÷ IR(KBr): 1765, 1668, 1598 cm"1 k ample C27 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-[2-(4hydroxy-phenyl)-5-methyl-[1,2,4]triazolo[1,5-a]pyrimidin-7-ylsulfanyl]- acetyl Amino]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodinm salt (1:1) Yield: 22.5% colorless lyophilisate MS(ISP): 634.4 (M+H)÷ IR(KBr): 1765, 1666, 1613 cm"1 « mple C28 (E)-(6R,7R)-3-(1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2-(4-methyl4H-[1,2,4]triazol-3-ylsulfanyl)-acetylamlno]-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 53.5% beige lyophilisate MS(ISP): 491.4 (M+H)÷ IR(KBr): 1764, 1673, 1615 cm1 l ample C29 (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-7- [2-(pyrimidin-2-ylsulfanyl)-acetylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene2-carboxylic acid sodium salt (1:1) Yield: 55.2% yellowish lyophilisate MS(ISP): 502.1 (M+H)÷ IR(KBr): 1762, 1664, 1614 cm 1 k -ple C30a Mixture of (E)-(6R,7R)-7-[(R)- and -[(S)-2-bromo-propionylamino]-3-(1cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt, see ex. Dla The following compounds were prepared according to Example C4 E ample C3( Mixture of (E)-(6R,7R)-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3ylidenemethyl)-8-oxo-7-[(R)- and -[(S)-2-(1H-pyrazolo[3,4-d]pyrimiin-4CA 02220188 1997-11-04 ylsulfanyl)-propionylamino]-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 56.2% yellow lyophilisate MS(ISP): 556.2 (M+H)÷ IR(KBr): 1764, 1666, 1621 cm« ample C31 Mixture of (E)-(6R,7R)-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3ylidenemethyl)-7-[(R)- and -[(S)-2-(4-methyl-4H-[1,2,4]tñazol-3-ylsulfanyl)- propionylamino]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 64.0% beige lyophilisate MS(ISP): 519.2 (M+H)÷ IR(KBr): 1764, 1667, 1619 cm1 ET mple C32 Mixture of (E)-(6R,7R)-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin-3ylidenemethyl)-8-oxo-7-[(R)- and-[(S)-2-(2,4,5-tñchloro-phenylsulfanyl)- propionylamino]-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (1:1) Yield: 60.2% colorless lyophilisate MS(ISP): 617.0 (M+H)÷ IR(KBr): 1760, 1670, 1618 cm1 Example C33 (E)-(6R,7R)-7-[2-[2-(3,4-Dihydroxy-phenyl)-5-methyl-[1,2,4]triazolo[1,5a]pyrimidin-7-ylsulfanyl]-acetylamlno]-3-[1-[ 1-[(3-fluoro-4-hydroxyphenylcarbamoyl)-methyl]-pyridin1-ium-ylmethyl]-2-oxo-pyrrolidin-3ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate (E)-(6R,7R)-7-(2-bromo-acetylamino)-3-[1-[ 1-[(3-fluoro-4-hydroxyphenylcarbamoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2-oxo-pyrrolidin-3ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate (see ex. Cla) (337.0 mg, 0.50 mmol) was dissolved in 4 ml N,Ndimethylformamíde and 4-[7-mercapto-5-methyl-s-triazolo[1,5-a]pyrimidin2-yl]pyrocatechol sodium sait (162.4 mg, 0.55 mmol) was added. After 6h the reaction mixture was poured on diethyl ether and the solid material was filtered. The brown powder was suspended in 8 ml water : acetonitrile (3 : 1), the pli was adjusted to 3 with 1M hydrochloric acid and the suspension was chromatographed on MCI gel (75-150 Mitsubishi Kasei Corporation) with a CA 02220188 1997-11-04 gradient of water : acetonitrile (4 : 1, 3 : 1, 2 : 1, 1 : 1 1 : 2, 1 : 3). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 12.0% brown lyophilisate MS(ISP): 868.4 (M+H)÷ IR(KBr): 1774, 1680, 1644 cm"1 mple C34a (E)-(6R,7R)-7-(2-Bromo-acetylamino)-3-[ 1-(4-hydroxy-benzyl)-2-oxo-pyrrolidin3-ylidenemethyl]-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2 carboxylic acid (E)-(6R,7R)-7m no-3-[1-(4-hydroxy-benzyl)-2-oxo-pyrrolidin-3ylidenemethyl]-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid (736.0 mg, 1.83 retool) was suspended in 20 ml dichloromethane and N,Obis(trimethylsilyl)-trifluoroacetamide (1.94 ml, 7.32 retool) was added. After 45 relu bromoacetyl bromide (167 ml, 1.92 retool) was added dropwise. After lh the reaction mixture was poured on 200 ml diethyl ether containing 160 ml water. After 15 min the solid material was collected by filtration, washed with diethyl ether and dried in high vacuum.

Yield: 84.1% yellowish solid MS(ISP): 522.3 (M+H)÷ IR(KBr): 1779, 1667, 1615 cm-1 E mple C34b (E)-(6R,7R)-7-[2-(4-Carboxy-phenylsulfanyl)-acetylamino]-3-[1-(4-hydroxybenzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct2-ene-2-carboxylic acid sodium salt (1:2) 4-Mercaptobenzoic acid disodium salt (83.0 mg, 0.42 mmol) was dissolved in 4 ml N,N-dimethylformamide and (E)-(6R,7R)-7-(2-bromo-acetylamino)-3-[1- (4-hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl]-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2 carboxylic acid (200.0 mg, 0.40 retool) was added in a single portion. After 2h the reaction mixture was poured on diethyl ether and the solid material was collected by filtration. The beige solid was suspended in water and the pli was adjusted to 7 with 1M sodium hydroxide solution. The resulting solution was chromatographed on MCI gel (75-150 Mitsubishi Kasei Corporation) with a gradient of water : acetonitrile (9 : 1, 8 :

2, 7 : 3). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 50.0% beige lyophilisate MS(ISP): 569.1 (M+H)÷ CA 02220188 1997-11-04 IR(Nujol): 1763, 1663, 1592 cm"1 The following compounds were prepared according to Example C34 Example C2 (E)-(6R,7R)-7-[2-[5-Carboxy-2-(3,4-dihydroxy-phenyl)-5-methyl- [1,2,4]tñazolo[1,5-a]pyrimidin-7-ylsulfauyl]-acetylamino]-3-[1-[1-[(3-fluoro-4hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-ylmethyl]-2-oxopyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate sodium salt (1:1) Yield: 20.0% beige lyophilisate MS(ISN): 896.3 (M-Na)" IR(Nujol): 1775, 1673, 1643 cm"1 - -ple C36 (E)-(6R,7R)-7-[2-(4-Carboxy-phenoxy)-acetylamino]-3-[1-[1-[(3-fluoro-4hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4-ylmethyl]-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate sodium salt (1:2) Yield: 20.0% brown lyophilisate MS(ISP): 748.4 (M+H)+ IR(Nujol): 1766, 1668, 1643 cm"1 mple C37 (E)-(6R,7R)-7-[2-[2-(3,4-Dihydroxy-phenyl)-5-methyl-[1,2,4]triazolo[1,5a]pyrimidin-7-yl sulfanyl]-acetylamino]-3-[1-(4-hydroxy-benzyl)-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid (E)-(6R,7R)-7-(2-bromo-acetylamino)-3-[ 1-(4-hydroxy-benzyl)-2-oxo-pyrrolidin3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid (see ex. C34a) (200.0 mg, 0.38 retool) was dissolved in 4 ml N,Ndimethylformamide and 4-[7-mercapto-5-methyl-s-triazolo[1,5-a]pyrimidin2-yl]pyrocatechol sodium salt (124.0 mg, 0.42 retool) was added. After 5h the reaction mixture was poured on diethyl ether and the suspension was filtered. The solid material was digerated with 10 ml ethyl acetate : water (1 :

1) for lh, filtered and dried in high vacuum.

Yield: 70.0% beige solid MS(ISP): 716.3 (M+H)÷ IR(KBr): 1774, 1670, 1596 cm1 CA 02220188 1997-11-04 The following compound was prepared according to Example C23 Example C38 (E)-(6R,7R)-7-[2-(4-Amino-phenylsulfanyl)-acetylamino]-3-(1-cyclopropyl-2oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylic acid sodium salt (1:1) Yield: 37.0% beige lyophilisate MS(ISP): 501.2 (M+H)÷ IR(KBr): 1782, 1623, 1599 cm"1 The following compounds were prepared according to Example C33 F, Ample C39 (E)-(6R,7R)-7-[2-(2,6-Dichloro-phenylsulfanyl)-acetylamino]-3-[1-[1-[(3-fluoro4-hydroxy-phenylcarbamoyl)-methyl]-pyñdin1-ium-i-ylmethyl]-2-oxopyrrolidin-3-ylidenemethyl]- 8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate Yield: 19.0% beige lyophilisate MS(ISP): 772.3 (M+H)÷ IRçKBr): 1768, 1663, 1643 cm-1 example CA0 (E)-(6R,7R)-7-[2-(3,5-Dichloro-phenylsulfanyl)-acetylam no]-3-[1-[1-[(3-fluoro4-hydroxy-phenylcarbamoyl)-methyl]-pyridin-l-ium-4-ylmethyl]-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2carboxylate Yield: 19.0% beige lyophilisate MS(ISP): 772.3 (M+H)÷ IR(KBr): 1769, 1677, 1643 cm1 Method D Example Dla Mixture of (E)-(6R,7R)-7-[(R)- and -[(S)-2-bromo-propionyl_am no]-3-(1cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt Br / NH COONa o CA 02220188 1997-11-04 A suspension of (E)-(6R,7R):7-amino-3-(1-cyclopropylmethyl-2-oxo-pyrrolidin3-ylidenemethyl)-8-oxo-5-thia1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (3.50 g, 10.0 retool) in 50 ml dichloromethane was treated with N-methyl-Ntrimethylsilyl-trifluoroacetamlde (4.40 ml, 24.0 retool). After 1 h a yellow solution was formed to which 2-bromo-propionyl bromide (1.30 ml, 12.0 rnmol) was added dropwise at O'C. The reaction mixture was stirred for rein at 0°C and for 3 h at room temperature and then concentrated in vacuo.

The oily residue was suspended in water and the pli was adjusted to 6.8 using 1 N sodium hydroxide solution. The resulting solution was purified by reversed-phase chromatography (RP-18 LiChroPrep gel, water : acetonitrile =1:0,4:1).

Yield: 2.80 g (55.0 %) -1 IR(KBr): 1765, 1667 cm MS(ISN): 482.1 (M-Na)÷ l mple Dlb Mixture of (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[(R)- and- [(S)-2-(2,5-dichloro-phenylsulfanyl)-propionylamino]-8oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid c, .,h_¢s.h COOH 0 A 0.05 N solution of a mixture of (E)-(6R,7R)-7-[(R)- and -[(S)-2-bromopropionylamino]- 3-( 1cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt in DMSO (500 ill) was treated with with a 0.05 N solution of 2,5dichlorobenzenethiol in DMSO (500 gl) at room temperature for 24 h.

MS(ISP): 599.2 (M+NHJ The following compounds were synthesized according to the procedure described above:

mpleD2 Mixture of (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[(R)- and- [(S)- [2-(1-methyl-lH-tetrazol-5-ylsulfanyl)-propionylamino]-8-oxo-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt CA 02220188 1997-11-04 N N/ N S COONa O MS(ISP): 537.3 (M+NH4 )÷ E ample 1)3 Mixture of (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3ylidenemethyl)-8-oxo-7-[(R)- and- [(S)- [2-(pyrimidin-2-ylsulfanyl)- propionylam no]-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid I N S I N H.

COON 0 MS(ISP): 533.2 (M+NH4 )+ k' mple 1)4 Mixture of (E)-(6R,7R)-3-(1-Cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[(R)- and- [(S)- [2-(4-methoxy-phenylsulfanyl)-propionylamino]-8oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid /O ì s NH _ COOH 0 MS(ISP): 561.2 (M+NH4 )+ CA 02220188 1997-11-04 Method E Example E1 (E)-(6R,7R)-3-[1-[ 1-[(4-Hydroxy-phenylcarbamoyl)-methyl]-pyridin1-ium-4ylmethyl]-2-oxo-pyrrolidin-3-ylidenemethyl)]-8-oxo-7-[2-(pyridin-l-ium1-yl)- acetyl_amino]-5-thia-l-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate bromide H _ +,,, 0 OH + N S N Nil Br COOO To a suspension of 260.0 mg (0.40 retool) (E)-(6R,7R)-7-amlno-3-[1-[1-[(4hydroxy-phenylcarbamoyl)-methyl]-pyñdin-l-ium-4-ylmethyl]-2-oxopyrrolidin-3-ylidenemethyl]-8-oxo-5-thia1-aza-bicyclo[4.2.0]oct-2-ene-2earboxylate tritluoroaeetate in 3 ml dichloromethane were added 212 ILl (0.80 retool) N,O-bis-(trimethylsilyl)-trifluoroacetamide. After a clear solution had formed, 65 gl (0.80 retool) pyridine and 35 IL1 (0.40 retool) bromoacetyl bromide were added and the reaction mixture was stirred for 6 h. To this solution 3 ml diethyl ether and 10 1 water were added. The precipitate was filtered off, dissolved in 250 l water" acetonitrile (3 • 2) and purified by reversed phase chromatography (RP-18 LiChroPrep gel, water " acetonitrile = 9 : 1). The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dñed.

Yield: 110.0 mg (45.3%) beige lyophilisate IR(KBr): 1768, 1682, 1635, 1574 cm"1 MS(ISP): 655.4 (M+H÷) CA 02220188 1997-11-04 where R1