IE57098B1 - Carbapenem process - Google Patents

Abstract

Disclosed is a new and more efficient process for producing carbapenem antibiotic derivatives having a 2- substituent of the general formula S-A-R<14> wherein A represents cyclopentylene, cyclohexylene or C2- C6 alkylene optionally substituted by one or more C1-C4 alkyl groups and R<14> represents a quaternized nitrogen-containing aromatic or non-aromatic heterocycle attached to A through a quaternary nitrogen atom. The carbapenems produced by this process are of the formula R<8> being hydrogen and R<1> being hydrogen or one of various defined organic groups or R<1> and R<8> together completing a ring and R<2> being hydrogen, an anion or a protecting group. The starting materials are of the formula and H-S-A-R<14> L being a leaving group and R<2> being a protecting group.

Description

The present invention is directed to a new process for Hu· preparation (»C varbap»*nom derivatives having a 2-nubstjtuent t l.hf formula -S-A-R*4 wherein A represents cyclope.nty lone , eye iohexylene or alkylene optionally substituted by one or more C^-C. alkyl groups and R represents a quaternized nitrogen-containing aromatic or non-aromatic heterocycle attached to A through a quaternary ni trogen atom.

The carbapenem derivatives prepared by the process of the present invention are disclosed and claimed by my colleagues Choung U. Kim and Peter F. Misco, Jr., in U.S. Patent Applications Serial Nos. 366,910, 471,379 and 389,652 filed, respectively, April 9, 1982, March 8, 1983 and June 18, 1982.

U.S. Application 366,910 and its continuation-in-part U.S. Application Serial No. 471,379, filed March 8, inR3, which continuation-in-part corresponds to West German Published latent Application 3,312,533, disclose preparation of carbapenem antibiotics of the formula 1 wlierHn R .is hydrogen and R is selected from the group consisting of hydrogen; substituted and unsuhstituted: alkyl, alkenyl and alkynyl, having from 1-10 carbon atoms; cvcloalkyl and cycloalky Iι alkyl, having 3-6 carbon atoms in'the cycloalkyl ring^and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl; and aralkynyl wherein the aryl moiety is Phenyl and the alipha’tic portion has 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and hetcrocyclylalky1 wherein the hetero atom or atoms in the above-named hoterocyclic moieties are selected from the group consisting ol; 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with said heterocyclic moieties have 1-6 carbon atoms; wherein the substituent or substituents relative to the above-named radicals are selected from the group consisting of C,-Cx alkyl optionally substituted by 1 0 amino, halo, hydroxy or carboxyl halo -OR Π 3 4 -OCNR R // 34 -CNJrR -NR3R4 /NR 4 -SO^NR R H 3 4 -NKCNR R J?CNR4-co2r3 =0 II 3 -OCR -sir o // g -SR* // 9 -SR* 1/ -CN -Γ,3 -i>so3n3 -OSO„R3 -NR SO2R -nr3c«nr4 k3 -NR3C02R4 -NO„ wherein, relative to the above-named substituents, the 3 4 groups R and R are independently selected from hydrogen; alkyl, alkenyl and alkynyl, having from 1-10 carbon atoms; cycloalkyi, cycloalkylalkyl and alkylcycloalkyl, having 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl wherein the aryl moiety is phenyl and the aliphatic portion has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl wherein the hetero atom or atoms in the above-named heterocyclic moieties are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with said heterocyclic moieties have 1-6 3 4 carbon atoms, or R and R taken together with the nitrogen to which at least one is attached may form a - or 6-membered nitrogen-containing heterocyclic ring; 3 R is as defined for R except that it may not be hydrogen 8 or wherein R and R taken together represent C2~^lO alkylidene or alkylidene substituted by hydroxy; A is cyclopentylene, cyclohexylene or ^2C6 alkylene optionally substituted by one or more C.-C. alkyl groups; 1 2 R is hydrogen, an anionic charge or a conventional readily removable carboxyl protecting group, providing that when R2 is hydrogen or a protecting group, there is also present a counter anion; and θ -N represents a substituted ox unsubstituted mono-, bi- or polycyclic aromatic heterocyclic radical containing at least one nitrogen in the ring and attached to A through a ring nitrogen, thereby forming a quaternary ammonium group; and pharmaceutically acceptable salts thereof by the process shown in the following reaction scheme: diphenyl chlorophosphate Ill S-A-OH ’ COOR HS-A-OK -> A = alkylene or C^-Cg cycloalkylene methanesulfonyl .chloride ---.----> Q) V «2 ^COOR2 ΙΑ Application 389,652 and its continuation-in-part application 499,690 filed June 7, 1983 disclose preparation of carbapenem antibiotics of the formula 1 '> wherein R is hydrogen and R is selected from the group consistinc of hydrogen; substituted and unsubstituted .* alkyl, alkenyl and alkynyl, having from 1-10 carbon atoms; cycloalkvl and cvcloalkylalkyl, having 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl wherein the aryl moiety is phenyl and the aliphatic portion has 1-6 carbon atoms; heteroaryl, heteroaralky1, heterocyclyl and heterocyclylalkyl wherein the hetero atom or atoms in the above-named heterocyclic moieties are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with said heterocyclic moieties have 1-6 carbon atoms; wherein the substituent or substituents relative to the above-named radicals are independently selected from the group consisting of C^-Cg alkyl optionally substituted by amino, halo, hydroxy or carboxyl halo -OR311 3 4 -OCNR R K 3 4 -CNRJR 4 -NRk NR' NR3R4 -SO2NR3R4 0 M 3 4 tNHCNR R r3cnr‘ -co2r; -OCR3 -SR3 -SR? -SR -CN -N, •oso3r oso2r3 •NR3SO2r' & -nr3onr4 -NR3C02R4 -N02 wherein, relative to the above-named substituents, the 3 4 groups R and R are independently selected from hydrogen; alkyl, alkenyl and alkynyl, having from 1-10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl, having 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl wherein the aryl moiety is phenyl and the aliphatic portion has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl wherein the hetero atom or atoms in the above-named heterocyclic moieties are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the. alkyl moieties associated with said heterocyclic moieties have 1-6 carbon atoms, or R3 and R4 taken together with the nitrogen to which at least one is attached may form a 5- or 69 membered nitrogen-containing heterocyclic ring; R is as 3 defined for R except that it may not be hydrogen; or 18 wherein R and R taken together represent alkylidene or C2~C10 alkylidene Substituted by hydroxy; is selected from the group consisting of substituted and unsubstituted: alkyl, alkenyl and alkynyl, having from 1-10 carbon atoms; cycloalkyl and cycloalkylalkyl, having 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl wherein the aryl moiety is phenyl and the «aliphatic portion has 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl wherein the hetero atom or atoms in the above-named heterocyclic moieties are selected from the group consisting of 1-4 * 30 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with said heterocyclic moieties have 1-6 carbon atoms; wherein the above-named radicals are optionally substituted by 1-3 substituents independently selected from: C^-Cg alkyl optionally substituted by ariino, fluoro, chloro, carboxyl, hydroxy or carbamoyl, fluoro, chloro or bromo; ί - OR ; -OCO,R3 5 -OCOR3 ί 3 4 -OCONR R ? -OSO^R3 ? -0X0' ; A -NR R ; R3CONR4~ Ϊ -nr3co«,r4 ϊ 3 4 -NR CONK R ; ~NR3SO,R4 ; μ* "SR3 ; <5 -S-R ; 0 -S-R ; -so3r3 r -CO2R3, ? 4 -CONRR ; -CN; or phenyl optionally substituted by 1-3 fluoro, chloro, bromo, C\-C- alkyl, -OR3, -NR3R4, A «Κ * 0 W J -SO,R , -CO,iC or -CONR R , wherein R , R-* toi R in such R substituents are as defined above or R3 may represent a divalent phenylene or C^-C^ alkylene group joined to the ring so as to form a bridged polycyclic group; A is cyclopentylene, or cvclohexvlene or C~-Cc alkylene optionally substituted by one 2 o 2 or more alkyl groups; R is hydrogen, an anionic charge or a conventional readily removable carboxyl protecting group, provid*> inq that when R-* is hydrogen or a protecting group, there is also present a counter ion; and I’i represents a substituted or unsubstituted mono-, bi- or polycyclic non-aromatic heterocyclic radical containing at least one nitrogen in the ring and attached to A through a ring nitrogen, thereby forming a quaternary ammonium group; and pharmaceutically acceptable salts thereof, by the process shown in the following reaction scheme: diphenyl chlorophosphate III v) S—A—OH COOK2' HS—A—OH -> A=alkylene or C5-Cg cycloalkylene roethanesulfonyl chloride -> IB To elaborate on the prior art scheme, starting material III is reacted in an inert organic solvent with diphenyl chlorophosphate in the presence of a base to give intermediate iv. Intermediate IV is then reacted with a mercaptan reagent of the formula HS-A-OH in an inert organic solvent and in the presence of a base to give intermediate V.

Intermediate V is then acylated with methanesulfonyl chloride in an inert organic solvent and in the presence of base to give intermediate VI which is reacted with a source of iodide ions in an inert organic solvent to give intermediate II.

Intermediate II is reacted with the desired amine in an inert organic solvent and in the presence of silver ion to produce the quaternized product I'A or I'B which may then be de15 blocked to give the corresponding de-blocked carbapenem of Formula IA or IE.

The above-described process has several disadvantages.

Thus, for example, the process involves several steps which advantageously could be reduced in number. The overall reaction 2.0 yield is also quite low and the quaternization step is performed on the entire carbapenem compound. It would be desirable to have a new process for producing compounds of Formula IA or IB which (1) involves fewer reaction steps, (2) gives higher yields, ΟΪ allows the quaternized amine to be formed first and then attached to the carbapenem nucleus at a later stage in the synthesis and (4) can be * used to more easily form quaternary amine products with a wide variety of amines, i.e. amines with steric hindrance and those with low pKfa values.

The present invention provides a novel process for preparation of carbapenem derivatives of the formula .8 wherein K is hydrogen hydrogen; substituted und R is selected from and unsubstituted: alkyl, alkenyl and alkynyl, having from 1-10 carbon atoms; cycloalkyl and cycloalkylalkyl, having 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl wherein the aryl moiety is phenyl and 10 the aliphatic portion has 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalky1 wherein the hetero atom or atoms in the above-named heterocyclic moieties are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with said hetero15 cyclic moieties have 1-6 carbon atoms; wherein the substituent or substituents relative to the above-named radicals are selected from C^-Οθ alkyl optionally substituted by amino halo, hydroxy or carboxyl halo -oS:nr3r' -$nr3r4 -nr3r4 NR' nr3r4 -S02NR3R4 -nhcnr3r4 J? R CNR -COgR Jr9 -CN -N ϊ. 7 -0303R3 -OSO-R3 -NITSOgR -nr3c=nr4 k3 -NR3C02R4 -NO2 wherein, relative to the above-named substituents, the 3 4 groups R and R are independently selected from hydrogen; alkyl, alkenyl and alkynyl, having from 1-10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl, having 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl wherein the aryl moiety is.phenyl and the aliphatic portion has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl wherein the hetero atom or atoms in the above-named heterocyclic moieties are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with said heterocyclic moieties have 1-6 carbon atoms, or R^ and R*4 taken together with the nitrogen to which at least one is attached may form a 5g or 6-membered nitrogen-containing heterocyclic ring; R 3 is as defined for R except that it may not be hydrogen; 8 or wherein R and R taken together represent alkylidene or C2"C10 alkyHdene substituted by hydroxy; A is cyclopentylene, cyclohexylene or C2"C6 alkylene optionally substituted by one or more C--C. alkyl groups; l 4 R is hydrogen, an anionic charge or a readily removable carboxyl protecting group, providing 2 ‘ that when R is hydrogen or a protecting group, there is also present a counter V' anion; and R^4 is a quaternized nitrogen-containing aromatic or non-aromatic heterocycle attached to A through a ring nitrogen, thereby forming a quaternary ammonium group, or a pharmaceutically acceptable salt thereof, which process comprises reacting an intermediate of the formula 2' wherein R and R are as defined above, R is a readily removable carboxyl protecting group and L is a leaving group,with a thiol compound of the formula HS-A-R14 Χθ VII Θ wherein A and R are as defined above and X is a counter anion j in an inert solvent and in the presence of base, to produce a carbapenem product of the formula I' 182' 14 Θ wherein R , R , R , A, R and X are as defined above and, if 2 ’ desired, removing the carboxyl protecting group R to give the corresponding de-blocked compound of formula I, or a pharmaceutically acceptable salt thereof. ί ίί Also provided by the present invention are intermediates of Formula VXI and processes for preparing such intermediates.

The carbapenem compounds of Formula I are 5 potent antibacterial agents or intermediates useful in the preparation of such agents.

The compounds of general Formula I above contain the carbapenem nucleus and may thus be named as l-carba-2-penem-3-carboxylic acid derivatives. Alternatively, the compounds may be considered to have the basic structure 4 and named as 7-axo-l~azabicyclo(3.2.0)hept-2-ene-2-carboxylic 15 acid derivatives. While the present invention includes compounds wherein the relative stereochemistry of the 5,6-protons is cis as well as trans, the preferred compounds have the 5R,6s (trans? stereochemistry as in the case of thienamycin.

The compounds of Formula I may be unsubstituted in 20 the 6-position or substituted by .substituent groups previously disclosed for other carbapenem derivatives. More specifically, 1 R may be hydrogen and R may be hydrogen or a non-hvdrogen substituent disclosed, for example, in European Patent Application 8 1 38,869 (see definition of R^). Alternatively, R and R taken o together may be c2"cxo alkylidene or C2-C1O alkV1-^dene substituted, for example, by hydroxy. 8 To elaborate on the definitions for R and R : (a) The aliphatic alkyl, alkenyl and "alkynyl groups may be straight or branched chain having 1-10 carbon atoms; preferred are 1-6, most preferably 1-4, carbon groups; when part of another substituent, e.g. as in cycloalkylalkyl, or heteroaralkyl or aralkenyl, the alkyl, alkenyl and alkynyl group preferably contains 1-6, most preferably 1-4, carbon atoms. (b) heteroaryl includes mono-, bi- and polycyclic aromatic if> heterocyclic groups containing 1-4 Ο, N or S atoms; preferred are 5- or 6- moml.M'red heterocyclic rings such as, for oxiinple, thienyl, furyl, thiadiazolvl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, tetrazclyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl and pyrazolyl. (c) heterocyclyl includes mono-, bi- and polycyclic saturated or unsaturated non-aromatic heterocyclic groups containing 1-4 0, N or S atoms; preferred are 5- or 6mcnixjred heterocyclic rings such as, for exanple, norpho].iny], piperoziny.l, piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrrolinylond pyrrolid.iny.1.. (d) halo includes chloro, bromo, fluoro and iodo and is preferably chloro or bromo.

The term read.i J.y removable carboxyl protecting group refers to a known ester group which has been employed to block a carboxyl group during the chemical reaction steps described below and which can be removed, if desired, by methods which do not result in any appreciable destruction of the remaining portion of the molecule, e.g. by chemical or enzymatic hydrolysis, treatment with chemical reducing agents under mild conditions, irradiation with ultraviolet light or catalytic hydrogenation. Examples of such ester protecting groups include benzhydryl, p-nitrobenzyl, 2-naphthylmethyl, allyl, benzyl, trichloroethyl, silyl such as trimethylsilyl, phenacyl, p-methoxybenzy1, acetonyl, o-nitrobenzyl, 4-pyridylmethyl and C,-Cc alkyl such as methyl, ethyl or t-butyl. Included within such protecting groups are those which are hydrolyzed under physiological conditions such as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethyl. Particularly advantageous carboxyl protecting groups are pnitrobenzyl which may be readily removed by catalytic hydrogenolysis and allyl which can be removed by Pd(P03)4-catalyzed reaction.

The pharmaceutically acceptable salts referred to above include the nontoxic acid addition salts, e.g. salts with mincr.il «icids such us, for cxainpl*', hydrodi.loric, hydrobromic, hydi Iodic, phosphoric and sulfuric and salts with organic acids such as maleic, acetic, citric, succinic, benzoic, tartaric, fumaric, mandelic, ascorbic, lactic, gluconic and malic. Compounds of Formula I in the form of acid addition salts may be written as R' = H or protecting group X ** θ Θ where X represents the acid anion. The counter anion X may be selected so as to provide pharmaceutically acceptable salts for therapeutic administration but, in the case of θ intermediate compounds of Formula I, X may also be a toxic 5 anion. In such a case the ion can be subsequently removed or substituted by a pharmaceutically acceptable anion to form active end product for therapeutic use. When acidic or basic 14 groups are present in the R group or on the quaternized R radical, the present invention may also include suitable Π) base or acid salts of these functional croups, e.g. acid addition salts in the case of a basic group and metal salts (e.g. sodium, potassium, calcium and aluminum), the ammonium salt and salts with nontoxic amines (e.g. trialkylamines, procaine dibenzylamine, 1-ephenamine, N-benzyl-S-phenethylamine tind N,N'15 dibenzylethylenediamine) in Lhe case of an acidic group.

Compounds of Formula I wherein R is hydrogen, an anionic charge or a physiologically hydrolyzable ester group together with pharmaceutically acceptable salts thereof are useful as antibacterial agents. The remaining compounds of Formula I are valuable intermediates which can be converted into the above-mentioned biologically active compounds.

A preferred embodiment of the present invention 1 comprises compounds of Formula I wherein R is hydrogen and R is hydrogen, CH^CH^- CH? OH CH CH-, OH I or CH3CHAmong this subclass, the preferred compounds are those in which is OH I CH^CH-, most preferably compounds having the absolute configuration 5R, 6S, 8R. ··» ·» 4,, (JO Another preferred embodiment comprises compounds of 1 8 Formula I in which R and R taken together form an alkylidene radical of the formula HOCH, C= The alkylene or of Formula I may be cycloalkylene radical A in the compounds cyclopentylene cyclohexylene or C2"C6 alkylene optionally substituted by one or more cj,"C4 alkyl substituents. Preferred A substituents are cyclopentylene, cyclohexylene or alkylene of the formula R I -c R12 I -cκ 1 k13 «12 R , R"" and R are each independently hydrogen or A preferred embodiment comprises those compounds of Formula I in which'substituent A is -CH^CH^·, in which R alkyl.

CH. or CH.CH2| CH.

In the case of certain compounds of Formula I having a cycloalkylene or branched alkylene A substituent, one or more additional assymmetric carbon atoms may be created which result in formation of diastereoisomers. The present invention includes mixtures of such diastereoisomers as well as the individual ourified diastereoisomers.

The quaternized R substituent may be an optionally substituted mono-, bi- or polycyclic aromatic or non-aromatic heterocyclic radical containing at least One nitrogen in the ring and attached to A through a ring nitrogen, thereby forming a quaternary ammonium group.

One preferred class of R substituents may be represented by the general formula which is meant to define a substituted or unsubstituted mono-, id bi- or polycyclic heteroaryl radical containing at least one nitrogen in the ring and attached to a carbon atom of substituent A through a ring nitrogen, thereby forming a quaternary ammonium group. The heteroaryl radical may be optionally substituted by such substituents as C^-C4 alkyl, C^-C4 alkyl substituted by hydroxy, amino, carboxy or halo, C^-Cg cycloalkyl, C^-C4 alkoxy, C1~C4 a^ylthio, amino, C^-C4 alkylamino, di(Cj_-C4 alkyl) amino, halo, Cj.~C4 alkanoylamino, C^-C4 alkanoyloxy, carboxy, -C-OC1~C4 alkyl, hydroxy, amidino, guanidino, trifluoromethyl, phenyl, phenyl substituted by one, two or three amino, halo, hydroxyl, trifluoromethyl, C^-C4 alkyl or C^-C4 alkoxy groups, heteroaryl and heteroaralkyl' in which the hetero atom or atoms in the above-named heterocyclic moieties arc selected from 1-4 0, N or S atoms and the alkyl moiety associated with hetero25 aralkyl has 1-6 carbon atoms.

The heteroaryl radical attached to substituent A is preferably a 5- or 6- membered aromatic heterocyclic radical containing a quaternized nitrogen atom (which is directly bonded to a carbon atom of the alkylene or cycloalkylene radical) and, Π) optionally, one or more additional hetero atoms selected from O, N or S. While, in general, any heteroaryl radical bonded to A Μ* «.fr via a quaternized nitrogen atom is found to produce biologically active carbapenem derivatives, a preferred embodiment comprises compounds in which represents a radical selected from p7 (a) R wherein R3, R& and R7 are independently selected from hydrogen; Cx-C4 alkyl; C1~C4 alkyl substituted by hydroxy, amino, carboxy or halo; C^-Cg cycloalkyl; C^-C4 alkoxy; C^-C4 alkylthio; amino; Ci-C4 alkylamino; di(C1-C4 alkyl)amino; halo; C1~C4 alkanoylamino; C^-C4 alkanoyloxy; carboxy; II ( -C-OC^-C4 alkyl; hydroxy, amadino; guanidino; trifluoromethyl; phenyl; phenyl substituted by one, two or three amino, halo, hydroxyl, trifluoromethyl, C^—alkyl or C1-C4 alkoxy groups; and heteroaryl and heteroaralkyl in which the hetero atom or atoms in the above-named heterocyclic moieties are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moiety associated with said heteroaralkyl moiety has 1-6 carbon atoms; or wherein two of R3, R6 or r' taken together may be a fused saturated carbocyclic ring, a fused aromatic carbocyclic ring, a fused saturated heterocyclic ring or a fused heteroaromatic ring, (b) ®ώ Π or optionally substituted on a carbon atom by one or more substituents independently selected from C^-C4 alkyl; C^-C4 alkyl substituted by hydroxy, amino, carboxy or halogen; C^-C^ cycloalkyl; C|'^4 alkoxy; C|-C4 alkylthio; amino; C^-C^ alkylamlno; di(C|-C4 alkyl) amino; halo; C^-C4 alkanoylamino; C^-C4 alkanoyloxy; O II carboxy; -C-OC^-C^ alkyl; hydroxy; amidino; guanidino; trifluoromethyl; phenyl; phenyl substituted by one, two or three amino, halo, hydroxyl, tri fluoromethyl, C^-C4 or ci~C4 ai^oxy groups; and heteroaryl or heteroaralkyl in which the heteroatom or atoms in the above-named heterocyclic moieties are selected from 1-4 oxygen, nitroqen or sulfur atoms and the alkyl moiety associated with said heteroaralkyl moiety has 1-6 carbon atoms, or optionally substituted so as to form a fused carbocyclic or heterocyclic ring; optionally substituted on a carbon atom by one or more substituents independently selected from C^~C4 alkyl; C^-C4 alkyl substituted by hydroxy, amino, carboxy or halogen; C^-C^ cycloalkyl; C1"C4 alkoxy; C^-C4 alkylthio; amino; C^-C4 alkylamino; di(C^-C4 alkyl)amino; halo; C^-C4 alkanoylamino; C^-C4 alkanoyloxy; carboxy; f? -C-OC^-C4 alkyl; hydroxy; amidino; guanidino; trifluoromethyl; phenyl; phenyl substituted by one, two or three amino, halo, hydroxyl, trifluoromethyl, C^-C^ alkyl or C^-C4 alkoxy groups; and heteroaryl or heteroaralkyl in which the hetero atom or atoms in the above-named heterocyclic moieties are selected from l-Λ oxygen, nitrogen or sulfur atoms and the alkyl moiety associated with said heteroaralkyl moiety has 1*6 carbon atoms, or optionally substituted so as to form a fused carbocyclic or heterocyclic ring; optionally substituted on a carbon atom by one or more substituents it) independently selected from alkyl; cy"c4 alkyl substituted by hydroxy, amino, carboxy or halogen; C^-Cg cycloalkyl; C^-C4 alkoxy; C1-C4 alkylthio; amino; C^-C^ alkylamino; di(C^-C4 alkyl) amino; halo; C^-C'4 alkanoylamino; C^-C4 alkanoyloxy; carboxy; II -C-OC^-C4 alkyl; hydroxy; amidino; guanidino; trifluoromethyl; phenyl; phenyl substituted by one, two or three amino, halo, hydroxyl, trifluoromethyl, C^-C4 alkyl or C^-C4 alkoxy groups; and heteroaryl or heteroaralkyl in which the hetero atom or atoms in the above-named heterocyclic moieties are selected from 1-4 oxygen, nitrogen or sulfur atoms and tho 2D alkyl moiety associated with said heteroaralkyl moiety has 1-6 carbon atoms, or optionally substituted so as to form a fused carbocyclic or heterocyclic ring; wherein X is 0, S or NR 'in which R is C^-C4 alkyl or phenyl, said radical being optionally substituted on a carbon atom by one or more substituents independently selected from C^-C4 alkyl; ciC4 alkyl substituted by hydroxy, amino, carboxy or halogen; C^-Cg cycloalkyl; C^-C^j alkoxy; alkylthio; amino? C^-C4 alkylamino; di(C^-C4 alkyl)amino; halo; Cj.-C4 alkanoylamino; C|-C4 alkanoyloxy; carboxy; II -C-OC^-C4 alkyl; hydroxy; amidino; guanidino; trifluoronethyl; phenyl; phenyl substituted by one, two or three amino, halo, hydroxyl, trifluoromethyl, ci"c4 alkyl or alkoxy groups; and heteroaryl or heteroaralkyl in which the hetero atom or atoms in the above-named heterocyclic moieties are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moiety associated with said heteroaralkyl moiety has 1-6 carbon atoms, or optionally substituted so as to form a fused carbocyclic or heterocyclic ring; e ΙΟ wherein X is Ο, S or NR in which R is C^-C^ alkyl or phenyl, said radical being optionally substituted on a carbon atom by one or more substituents independently selected from C^-C^ alkyl; C^-C^ alkyl substituted by hydroxy, amino, carboxy or halogen; C^-C^ cycloalkyl; C^-C^ alkoxy; C^-C4 alkylthio; amino; C^-C^ alkylamino; di(C^-C^ alkyl)amino; halo; C^-C^ alkanoylarnino C^-C^ alkanovloxy; carboxy; II -C-OC^-C^ alkyl; hydroxy;· amidino; guanidino; trifluoromethyl; phenyl; phenyl substituted by one, two or three amino, halo, hydroxyl, trifluoromethyl, C^-C^ alkyl or C^-C^ alkoxy groups; and heteroaryl or heteroaralkyl in which the hetero atom or atoms in the above-named heterocyclic moieties oxygen nitrogen are selected from 1-4 or sulfur atoms and the alkyl moiety associated with said heteroaralkyl moiety has 1-6 carbon atoms; and N — N— I \ N^N-R N =* N e/ \ or N-N-R n \ wherein R is alkyl or phenyl, said radical being optionally substituted on the carbon atom by a substituent selected from C1-C4 alkyl; C^-C^ alkyl substituted by hydroxy; amino, carboxy or halcce C^-Cg cycloalkyl; C]_"C4 alkoxy; ciC4 alkylthio; amino; C1-C4 alkylamino; di(C^-C4 alkyl) amino; cj>*c4 alkanoylamino; carboxy; -C-OC^-C4 alkyl; hydroxy; amidino; guanidino; trifluoromethyl; phenyl, phenyl substituted by one, two or three amino, halo, hydroxyl, trifluoromethyl, C^-C4 alkyl or C^-C4 alkoxy groups; and heteroaryl or heteroaralkyl in which the hetero atom or atoms in the above-named heterocyclic moieties are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moiety associated with said heteroaralkyl moiety has 1-6 carbon atoms.

Within the above subclass, the preferred compounds are those in which substituent A is , / \ or CH J z OH^CH- and wherein either (a) R- and R taken together represent CH HOCHc= CH 1 or (b) R is hydrogen and R represents hydrogen, CH^CH^-r Ah OH OH CHor CH3CH· CH CH θ Particularly preferred are the compounds wherein R is hydrogen and is OH CH^H-, preferably compounds having the absolute configuration 5R, 65, 8R. :u A particularly preferred embodiment of the present invention comprises preparation of compounds wherein N J represents a radical of the formula in which R^, and R7 are independently selected from UyErouen, Cj-C4 alkyl, alkoxy, c1"c4 alkyl substituted by a hydroxy group, C1-C4 alkylthio, amino, carboxy and carbamoyl. Within this subclass, the preferred compounds are or those wherein substituent A is -CH-CH -, -CHCHn2 2 | 2 CH.

CH and wherein either (a) R and R taken together represent HOCHCH 1 or (b) R is hydrogen and R represents hydrogen, CH^CH^CH3 oh OH CH CHC- or CH3CH· CH and R^ is g Particularly preferred are the compounds wherein R is hydrogen £-. preferably compounds having the absolute configuration 5R, 6S, 8R. 7* ‘Ί aj *« A most preferred embodiment of the present invention comprises preparation of compounds wherein represents a radical of the formula 6 I I) .7 r’ in which R^z r® and are independently selected from hydrogen, C|C4 ‘^kyl, C l 4 η1^οχγ, C'iC4 alkyl substituted by a hydroxy group, C^-C^ alkylthio and amino Within this subclass, the preferred compounds are those wherein substituent A is -CH^CH^-, - or -CHnCH2/ CH-. \_/ CH1 4 8 ' J ‘ and wherein either (a) R and R taken together represent HOCH CH c= J 1 or (b) R is hydrogen and R represents hydrogen, CH^CH^· CHOH I cOH I or CH3CHCH CH Particularly preferred are the compounds wherein R is hydrogen and r1 is CH preferably compounds having the absolute configuration 5R, 6S, 8R.

Another preferred embodiment of the present invention 25 comprises preparation of compounds wherein represents a radical of the formula —sch3 /N-ch3 substituent A is ' Within this subclass, the preferred compounds are those wherein CH1 CH or -CH-CH2| 8 and wherein either (a) R and R taken together represent HOCH.

CH3 1 or (b) R is hydrogen and R represents hydrogen, CH^CH2 CH, Λ CH OH OH CH CH* Cor CH3CHg Particularly preferred are the compounds wherein R is hydrogen and R^ is OH I CH^CH-, preferably compounds having the absolute configuration 5R, 6S, 8R.

Another preferred embodiment of the present invention comprises preparation of compounds wherein witftm tms suociass, tne represents a pyridinium radical preferred compounds are those wherein substituent A is -CHCH^ CH. -ch2ch2or -CU2CH- and wherein either (a) CH18 J R and R taken together represent hoch7 CH ο »* c= 1 or (b) R is hydrogen and R represents hydrogen, CH ch3 OH OH CHCor CH3CHCH Particularly preferred are the compounds wherein R is hydrogen and R^ is OH ^H-, configuration 5R, 6S, 8R.

CH3CH-, preferably compounds having the absolute A most preferred embodiment of the present invention comprises the preparation of compounds of the formula wherein represents (Π) -CHCH.

I CH(12} R or S diastereoisomers (13) or (14) OR,R or S ,S diastereoisomers at two assymmetric carbons of the cyclohexyl group and R is hydroqen, an anionic charge or a conventional read ily removable carboxyl protecting group, providing that when K Is hydrogen or a protecting group, there is also present a counter anion, and pharmaceutically acceptable acid addition salts thereof.

Another preferred class of quaternized R substituents may be represented by the general formula where R1^ represents either (a) an optionally substituted 10 aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, aryl, araliphatic, heteroaryl, heteroaraliphatic, heterocyclyl or heterocyclyl-aliphatic radical or (b) a divalent phenylene or C1~C4 alkylene group joined to the ring so as to form a bridged polycyclic group. Thus R may be selected from substituted and unsubstituted: alkyl, alkenyl and alkynyl, having from 1-10 carbon atoms; cycloalkyl and cycloalkylalky1, having 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkeny] and aralkynyl wherein the aryl moiety is phenyl and the aliphatic portion has 1-6 carbon atoms; hetcroaryl, heteroalkyl, heterocyclyl and heterocyclylalkyl wherein the hetero atom or atoms in the abovenamed heterocyclic moieties are selected from 1-4 oxygen, nitrogen or sulfur atoms and the cilkyl moieties associated with said heterocyclic moieties have 1-6 carbon atoms; wherein the above-named R16 * * * 20 * * * * 25 radicals are optionally Substituted by 1-3 substituents independently selected from: ?.o Cl"C6 a^yl optionally substituted by amino, fluoro, chloro, carboxyl, hydroxy or carbamoyl; fluoro, chloro or bromo; -OR3 ; -OCO2R3 ; -OCOR3 j -oconr3r4 ι ~OSO2R3 ; -OXO'; -NR3R4 ; ' 4 R CONR ~ ; 4 -NR CO2a ; ~nr3conr3r4 ,-nr3so2x4. ? -SR3 ; *r a -S~R* ; Q ? x* 9 -S-SC 7 SO3R3 f -CO, a3 ; 4 -CONICIT ; -CN; or phenyl optionally substituted by 1-3 fluoro, chloro, bromo, C^~Cg alkyl, -OR3, ~NR3r\ -SO^R3, -C02R3or -CONR3R4, wherein R3 , R4 and r9 ia such’substituents are as defined above; or may represent a divalent phenylene or alkylene group joined to the ring so a to form a bridged polycyclic group and represents a substituted or unsubstituted mono-, bi- or polycyclic non-aromatic (which may be fused to another aromatic or nonaromatic ring) heterocyclic radical containing at least one nitrogen in the ring and attached to A through a ring nitrogen, thereby in forming a quaternary ammonium group. The heterocyclic radical may be saturated or unsaturated (with 1-2 double bonds) and may contain up to two additional hetero atoms in addition to the quaternary nitrogen, such additional hetero atoms being selected from 0, S(O) , N, NR15 or NR17R18 wherein m is 0, 1 or 2, R15 is m t'> hydrogen, optionally substituted C.,-C- alkyl or optionally substituted 18 .-.o phenyl and R and R are each independently optionally substituted C1*C6 alkyl or optionally substituted phenyl.

In a preferred embodiment represents a non-aromatic 4-7 membered, preferably 5- or 6membered, N-containing heterocyclic ring containing Q-2 double bonds and 0-2 additional hetero-atoms selected from 0, S(0) , N, 17 18 15 m NR or NR* R wherein m is 0, 1 or 2, R is hydrogen, C,-C-alkyl η V optionally substituted by 1-2 substituents indeoendentlv selected 3 3 4 3 * /5 from -OR , -NR R , -C0-.R , oxo, phenyl, fluoro, chloro, bromo, 3 4* -SO^R and -CONR R or phenyl optionally substituted by 1-3 substituents independently selected from C^-C^ alkyl, -OR3, -NR3R4, fluoro, chloro, bromo, -SO3R3, -CO2R2 and -CONR3R4 and R17 and R13 are each independently C.-C- alkyl optionally substituted by id a . _ JO 1-2 substituents independently selected from -OR , -NR Rq, -CO^V, oxo, phenyl, fluoro, chloro, bromo, -SO3R3 and -CONR3R4 or phenyl 3U optionally substituted by 1-3 substituents independently selected from C,-C„ alkyl, -OR3, -NR3R4, fluoro, chloro, bromo, -SO R , -1 6 . a *15 -CO2Rz and -CONRk , wherein R and R in such heterocyclic NR NR1*R18 groups are as defined above in connection with the R substituent. In such preferred embodiment the or ring may be optionally substituted by 1-3 substituents independently selected from (a) C.-C- alkyl optionally substituted by 1-2 substituents Ao, 34 3 4 34 334 -OCOR , -OCONR R, OXO; -NR R , -NR COR , -NR CONR R , -NK3SO2R4, -SR3, -SO3R3, -C0,R3 and -CONR3R4; (b) C--C- alkenyl optionally substituted by 1-2 substituents * b 3 independently selected from fluoro, chloro, bromo, -OR , ,r> -OCOR3, -OCONR3R4, OXO, -NR3R‘i, -NR3COR4 , -NR3CONR3R4, -NR3SO2R4, -SR3, -SO3R3, -COjR3 and -CONR3R4; (c) C2~Cg alkynyl optionally substituted by 1-2 substituents independently selected from fluoro, chloro, bromo, -OR3, -OCOR3, -oconr3r4, oxo, -nr3r4, -nr3cor4, -nr3conr3r4, -NR3S02R4, -SR3, -SO3R3, -COjR3 and -CONR3R4; (d) C3~C5 cycloalkyl optionally substituted by 1-2 substituents independently selected from fluoro, chloro, bromo, -OR3, -OCOR3, -OCONR3R4, oxo, -nr3r4, -NR3COR4, -NR3CONR3R4, -NR3SO2R4, -SR3, -SO3R3, -CO,R3 and -CONR3R4; (e) cycloalkylalkyl having 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moiety, optionally substituted by 1-2 substituents independently selected from fluoro, chloro, bromo, -OR3, -OCOR3, -OCONR3R4, oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -NR3SO,r‘4, -SR3, -SO3R3, -CO,R3 and JO -CONR3R4; (fj heteroaryl wherein the hetero atom or atoms are selected from 1-4 oxygen, nitrogen or sulfur atoms, optionally substituted by 1-2 substituents 3 independently selected from fluoro, chloro, bromo, -OR f . -ocor3, -oconr3r4, OXO, -nr3r4, -nr3cor4, -nr3conr3r4, "NR35O2R4, -SR3, -SOjR3, -CO2R3 and -CONR3R4; preferred heteroaryl radicals are 5- or 6-merobered aromatic heterocyclic. rings; (g) heteroaralkyl - wherein the hetero atom or atoms are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moiety has 1-6 carbon atoms, optionally substituted by 1-2 substituents independently selected from j 3 34 fluoro, chloro, bromo, -OR , -OCOR , -OCONR R , oxo, 34 334 34 3 3 -NR R , -NR COR , -NR CQNR R , -NR SO,R , -SR , -SO,R , -CO,R and -CONR R4; preferred heteroaralkyl are those in which the heteroaryl radical is a 5- or 6-membered aromatic heterocyclic ring and the alkyl moiety has 1-2 carbon atoms; (h) heterocyclyl wherein the hetero atom or atoms are selected from 1-4 oxygen, nitrogen or sulfur atoms, optionally substituted by 1-2 substituents independently selected from fluoro, chloro, bromo, -OR3, -OCOR3, -OCONR3?4, OXO, -nr3r4, -nr3cor4, -nr3conr3r4, -NR3SO2R4, -SR3, -SO3R3, -C02R3 and -CONR3R4; preferred heterocyclyl are 5- or 6-membered saturated or unsaturated rings ? (i) heterocyclylalkyl wherein the hetero atom or atoms are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moiety has 1-6 carbon atoms, l() optionally substituted by 1-2 substituents independently 3 3 4 selected from fluoro, chloro, bromo, -OR , -OCOR , -OCONR R 34 3 34 3 4 3 ’3 0X0, -NR R , -NR COR , -NRJCONR R , -NR SO,R , -SR , -SO,? , 3 4 4' J -CO^R and -CONR R ; preferred heterocyclylalkyl are those in which the heterocyclyl moiety is a 5- or 6-membered saturated or unsaturated ring; .) (j) fluoro, chloro or bromo; (k) "OR3 ; (l) -OC02R3 ; (ra) -OCOR3 ; (n) -OCONR3R4 ; (o) -OSOjR3 ; (p) oxo ; (q) -NR3R4 ; (r) R3CONR4-; (s) -NR3CO2R4 ; (t) -NRJCONR3R4 ; (u) -NR3SO2R4 ; (v) -SR3 ; (W) 0 -Τ' 9 -S-R ; (x) 0 0 Λ Q -S-R ; (y) -SO3R3 ; (=) -CO2R3 ; (aa) -CONR3R4 ; (bb) -CN; or (cc) phenyl optionally substituted by 1-3 fluoro, chloro, bromo C^-C. alkyl, -OR3, -NR3R4, -SO3R3, -COjH3 or -CONR3R4. 4 Q The R , R and R* substituents mentioned above are as defined i connection with The ring as defined above is a non-aromatic heterocycle group.

This ring, however, may be fused to another ring which may be a saturated or unsaturated· carbocyclic ring, preferably a C^-C? carbocyclic ring, a phenyl ring, a 4-7 membered heterocyclic {saturated or unsaturated) ring containing 1-3 hetero atoms selected from 0, N, S (0)^, NR^, Or NR^R^ or a 5-6 membered heteroaromatic ring containing 1-3 hetero atoms selected 0, Ξ(0) 15 17 19 15 17 19 N, NR or NR R in which m, R , R and R are as defined above.

The R^S substituent of the non-aromatic R^4 radical may be either (a) an optionally substituted C^-Cg alkyl, C2~C1O alkenyl, C2C10 alkynyl, C^-Cg cycloalkyl, C^-Cg cycloalkylC^-Cg alkyl, phenyl, phenyl-C^-Cg alkyl, phenyl-C^'Cg alkenyl, phenyl-C^-C. alkynyl, heteroaryl, heteroaralkyl in which the alkyl moiety has 1-6 carbon atoms, heterocyclyl or heterocyclylalkvl in which the alkyl moiety has 1-6 carbon atoms or (b) a divalent phenylene or C^-C^ alkylene group joined to the ring so as to form a bridged ring polycyclic group, e.g. a cuinuclidine group. The heteroaryl (or heteroaryl portion of heteroaralkyl) substituent may be a mono-, bi- or polycyclic aromatic heterocyclic group containing 1-4 0, N or S atoms; preferred are 5- or 6-membered heterocyclic rings such as thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolvl, thiazolyl, imidazolyl, isoxazolyl, tetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl and pyrazolyJL. The heterocyclyl (or heterocyclyl portion of heterocyclylalkyl) substituent may be a mono-, bi- or polycyclic saturated or unsaturated πθη-aromatic heterocyclic group containing 1-4 0, N or S atoms; preferred are 5- or 6-membered heterocyclic rings such as morpholinyl, piperazinyl, piperidyl, pyrazolinyl, pvra2olidinyl, imidazolinyl, imida2olidinyl, pyrrolinyl and pvrrolidinyl.

In the case where the R1’6 substituent is an alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalky.l., phenyl,_ phenylalkyl, phenylalkenyl, phenylalkynyl, heteroaryl, heteroaralkyl, heterocyclyl or heterocyclylalky1 group, such groups may he optionally substituted by 1-3 substituents independently selected from: yl optionally substituted by, preferably 1-3, chloro, carboxyl, hydroxy or carbamoyl (a) C^-Cg alkyl amino, fluo, groups ; (b) fluoro, chi' (cl -OR3 ; (d) -OCOjR3 ; (e) -OCOR3 ; (f) -OCONR3r'5 ; (g) -OSOjR3 ; (h) -oxo ; (i) -NR3R4 ; (j) R3CONR4- ; (k) 3 4 -N« CO,R ; 3 ; 4 (1) -NR CONR R ΪΟ 4 (η) -NR3SO2S4 ; (η) -SR3 ; (ο) -SOR9 ; (ρ) -SO2R9 ; (q) -SO3R3 ; (r) -CO2R3 ; (s) -CONR3R4 ; (t) -CN ; or (u) phenyl optionally substituted by 1-3 substituents in-.. dependently selected from fluoro, chloro, bromo, C^-Cg alkyl, -OR3, -NR3R4, -SO^R3, -COjR3 or -CONR3R4, wherein, relative to the above-named substituents, the groups R3 and R4 are independently selected from hydrogen; alkyl, alkenyl and alkynyl, having 1-10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl, having 3-6 carbon atoms in the cycloalkyl ring and 1-6* carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl wherein the aryl moiety is phenyl and the aliphatic portion has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl wherein the heteroaryl and heterocyclyl group or portion of a group is as defined above for R^3 and the alkyl moieties associated with said heterocyclic moieties have 1-6 3 4 carbon atoms; or R and R taken together with the nitroce to which at least one is attached may form a 5- or 6-membered nitrogen-containing heterocyclic (as defined 9 3 above for R ) ring;' and R is as defined above for R except that it may not be hydrogen. A most preferred R^ substituent is C^-Cg alkyl, especially methyl.

Xn the case where R^6 is a divalent phenylene or C^_C6 alkylene group,, such group is bonded to another atom of the ring so as to form a bridged polycyclic ring, e.g. a quatemiced cuinuclidine ring of the formula A particularly preferred embodiment of the present invention comprises preparation of compounds of Formula I wherein <3 CH' wherein Y is hydrogen, C^Cg alkyl,'hydroxy, -SCj-Cg alkyl, carboxyl, carbamoyl, chloro, bromo, iodo/ fluoro or phenyl. Within this subclass, the preferred compounds are those wherein A is -CHCH I 1 CH.

, -CHjCHCH. or (CH2)n- in which n is 2, 3 or 4, more preferably those in which λ is -CH2CH2-, -CH2CH2CH2-, -chch2- , / \ or -ch2chI CH 2/ CH >-' ^"3 and most preferably those in which A is -CH-CH--, and wherein eithe: 18 * a (a) R and R taken together represent HOCH 0 ci£ c« 1 or (b) R is hydrogen and R represents hydrogen, C3,CZy J => C3 C3 CE oa CH^CH• 8 Particularly preferred are the compounds wherein R is hydrogen and is OH I ca3caespecially compounds having the absolute configuration 5R, SS, 8R.

A still more preferred invention comprises preparation Of embodiment of the present compounds of Formula I wherein CH CH \ΓΛ CH -\5 CH CH Θ >«3 CH NH or -\ί N ® \_/ ^CH jYY within this preferred subclass, the preferred compounds are those wherein A is -CHCH-I 2 CH3 CH-CH- or & I CH, -(CH^J^- in which n is 2, 3 or 4, more preferably those in which A is -CH-CH--, -CH-CH-CH--, ~CHCH«· 2 2 2 2 | 2 ch3 or -CH-CH- , and 2l CH, most preferably those in which A is -CH-CH-~, and wherein either 18 2 2 (a) R and R taken together represent HOCH. ch3 1 or (b) R is hydrogen and R represents hydrogen, CHjCSp CT3X.

CHCH3 oh OH I or CH3CH~ CH Particularly preferred are the compounds wherein R is * hydrogen and R is OH C33CH· , especially compounds having the absolute configuration SR, SS, 8R.

A still more preferred embodiment of the present invention comprises preparation of compounds of Formula I wherein -Ό <1 8 represents in which Y is hydrogen, ct"cg alkyl, hydroxy, -S-C^-Cg alkyl, carboxyl, carbamoyl, chloro, bromo, iodo, fluoro or phenyl.

Within this preferred subclass, the preferred compounds are those wherein A is -(CH.,)- in which n is 2, 3 or 4, most 2 n preferably those in which A is -C39CH9* and wherein either 4 2 (a) R and R taken together represent :och, c= CH. or (b) R is hydrogen and .X* represents hydrogen, CH CH CH.

CKCS OH 3^1 S' or C^CH8 Particularly preferred are the compounds wherein R is hydrogen and rA is OH I CH3CH", especially compounds, having the absolute configuration 5Rf 6S, 8R. <30 A most preferred embodiment of the present ir.ventio: comprises preparation of compounds of Formula I wherein Ό I represents Within this preferred subclass, the preferred compounds art those wherein A is -(C3 )- in which n is 2, 3 or 4, most &> ii preferably those in which A is -CH-θ’ - and wherein either 18 <& a (a) R and R taken together represent HOCH CH. or (b) R is hydrogen and R* represents hydrogen, CH^CH^CH CH, ?S or CH^CH' ca CH.

I r> g Particularly preferred are the compounds wherein R is hydrogen and R^ is OH I CH3CH", especially compounds having the absolute configuration SR, 6S, 8R.

A most preferred embodiment of the present invention comprises preparation of the compounds of the formula (both a- arid $diastereoisoroers) and R3 is hydrogen, an anionic charge or a conventional readily 2 removable carboxyl protecting group, providing that when R is hydrogen or a protecting group, there is also present a counter ion, and pharmaceutically acceptable acid addition salts thereof.

It will be appreciated that certain products within the scope of formula I may be formed as optical isomers as well as epimeric mixtures thereof. It is intended that the present invention include within its scope all such .optical isomers and epimeric mixtures, For example, when the 6-substituent is hydroxyethyl·, such substituent nay be in either the R or-S configuration and the resulting isomers as well as epimeric mixtures thereof are encompassed by the present invention.

The process of the present invention utilizes the intermediate of the formula R8 H 'COQR' IV which has been disclosed, for example, in European Patent Application 38,869 and which may be prepared by the general methods described therein. L represents a conventional leaving group (defined as X in European Patent Application 38,869) such as chloro, bromo, iodo, benzenesulfonyIoxy, p-toluenesulfonyIoxy, p-ni troben2en R8 H III _8 n2 R and R wherein R' acylating agent R -L. are as defined above with a suitable The preferred intermediate IV where L is diphenoxyphosphinyloxy may be prepared by reacting keto ester III in an inert organic solvent such as methylene chloride, acetonitrile or dimethylformamide with about an equimolar amount of diphenyl chlorophosphate in the presence of a base such as, for exanple, diisopropylethylamine, triethylamine, 4-dimethylaminopyridine at a temperature of Irom about -20°C to +40°C, most preferably at about 0°C. Intermediate IV may be isolated, if 25 desired, but is conveniently used as the starting material for the process of the present invention without isolation or purification.

In the present process, reacted with a quaternary amine carbapenem intermediate iv is thiol compound of the formula HS—Λ—R VII wherein A is cyclopentylene, cyclohexylene or C^-C^ alkylene optionally substituted by one or more C^-C^ alkyl groups, most preferably cyclopentylene, cyclohexylene or in which R R I -C· Rl2 and R13 R12 I -C" are each independently hydrogen c: Θ C^-C^ alkyl, X is a counter anion associated with a strong acid such as Cl , Br , CK^SO^ , CF .,50., or CF3S°3 CK /, ' and p14 is a quaternized nitrogen-containing aromatic or non-aromatic heterocycle as defined above. The reaction is carried out in an inert solvent such as acetonitrile, acetonitrile-dimethylformamide, Γ, tetrahyarofuran, tetrahvdrofuran-H^O, acetonitrile-K20 or acetone in the presence of base. The nature of the base is not critical.

Best results, however, have been obtained when a non-nucleophilic tertiary amine base such as diisopropylethylamine, 1,8-diazabicyclo[5.4.OJundec-7-ene, 1,5-diazabicyclo[4.3.0 J non-5-ene or a tri(C^25 C ) alkvlamine such as triethylamine, tributylamine or tripropylamine is employed. Reaction of intermediate IV with thiol VII may be carried out over a wide temperature range, e.g. -15°C up to room temperature, but is preferably done at a temperature in the range of from -15°C to +15°C, most preferably at around 0°C.

The carbapenem product produced by reaction of the quaternary amine thiol VII with intermediate IV will have a g θ counter anion associated with it [e.g. (C^H^O^PC^ · Cl or -anion associated with the quaternary thiol) which may at this stage be substituted by a different counter anion, e.g. one which is more pharmaceutically acceptable, by conventional procedures. Alternatively, the counter anion may be removed during the subsequent de-blocking step. Where the quaternized carbapenem compound and counter anion form an insoluble product, the product may crystallize out as it is formed and be collected pure by filtration.

Following formation of the desired carbapenem product, the carboxyl protecting group R of Compound I1 may be optionally removed by conventional procedures such as solvolysis, chemical reduction or hydrogenation. Where a protecting group such as ρ-nitrobenzyl, benzyl, benzhydryl or 2-naphthylmethy1 is used which can be removed by catalytic hydrogenation, intermediate 1' in a suitable solvent such as, for exanple, dioxanc-water-ethanol, tetral^drofurandiethylether-buffer or tetrahydrofuran-aqueous dipotussium hydrogen 15 phosphate-isopropanol may be treated under a hydrogen pressure of from 1 to 4 atmospheres in the presence of a hydrogenation catalyst such us, for example, palladium on charcoal, palladium hydroxide or platinum oxide at a t emperature of from 0 to 50°C for from about 2* 0.24 to 4 hours. When R is a group such as o-nilrobenzyl, pliotolyrus 20 may also be used for deblocking. Protecting groups such as 2,2,2-trichloroethyl may be removed by mild zinc reduction. The allyl protecting group may be removed by using a catalyst comprising a mixture of a palladium compound and triphenyl phosphine in a suitable aprotic solvent such as tetrahydroruran, methylene chloride or diethyl ether. Similarly/ other conventional carboxyl protecting groups may be removed by methods known to those skilled in the art. Finally, as mentioned above, compounds of formula 1' 2’ where R is a physiologically hydrolyzable ester such as acetoxymethyl, phthalidyl, indanyl, pivaloyloxymethyl, methoxymethyl, etc. may be administered directly to the host without de-blocking since such esters are hydrolyzed in vivo under physiological conditions. 8 It will be understood that where the R and/or R 14 substituent or the quaternized nucleophile R attached to substituent A contain a functional group which might interfere with the intended course of reaction, such group may be protected by a conventional blocking group and then subsequently de-blocked to regenerate the desired functional croup. Suitable blocking groups and procedures for intro40 ducing and removing such croups are well known to those skilled in the art.

As in the case of other g-lactam antibiotics, compounds of general Formula I may be converted by known procedures to pharmaceutically acceptable salts which, for purposes of the present invention, are substantially equivalent to the non-salted compounds. Thus, for example, one may dissolve a compound of 2 Formula I wherein R is an anionic charge in a suitable inert solvent and then add an equivalent of a pharmaceutically acceptable acid. The desired acid addition salt may be recovered by conventional procedures, e.g. solvent precipitation, lyophilisation, etc. Where other basic or acidic functional groups are present in the compound of Formula I, pharmaceutically acceptable base addition salts and acid addition salts may be similarly prepared by known methods.

A compound of Formula X where R is hydrogen or an anionic charge, or a pharmaceutically acceptable salt thereof may also be converted by conventional procedures to a corresponding 2 compound where R is a physiologically hydrolyzable ester group, or 2 a compound of Formula I wherein R xs a conventional carboxyl .’0 protecting group may be converted to the corresponding compound where R is hydrogen, an anionic charge or a physiologically hydrolyzable ester group, or a pharmaceutically acceptable salt thereof.

Certain of the thiol intermediates of Formula VII may be prepared, for example, by reacting a sulfide of the formula VIIIc wherein R^, R^, R^3 and are each independently hydrogen or Cn-C4 alkyl with a heteroaromatic amine (as defined above) of the formula or a non-aromatic the formula heterocyclic amine (as defined above) of r> and a strong acid. The reaction may be carried out in the presence or absence of an inert organic solvent which is preferably a non-polar organic solvent such as, for example, methylene chloride, benzene, xylene or toluene. Where the amine and sulfide reagents are liquids or where a solid amine is soluble in a liquid sulfide reagent, it is preferred to carry out the reaction without use of an additional solvent.

The particular strong acid used in the reaction is not critical and may be, for example, such strong inorganic or organic acids as, for example, hydrochloric, hydrobromic, methane1 r> sulfonic, p-toluenesulfonic and trif luoromethanesulfonic.

Formation of the quaternary amine thiol intermediate VII may be carried out at a temperature in the range of from -20°G to 100°C. Preferred temperatures are generally in the range of about 50-70eC.

The sulfide reagent, aromatic amine and acid are preferably employed so that the sulfide and acid are used in approximately equimolar amounts with the amine being used in excess, e.g, two to three moles of amine per mole of sulfide or acid, The quaternary amine thiol intermediate will have a counter anion associated with it which will be determined by the particular acid employed. It is, of course, possible to substitute at this point a different counter anion by conventional procedures for use in the subsequent reaction with carbapenem intermediate IV. 5G The carbapenem derivatives of general Formula I wherein R is hydrogen, an anionic charge or a physiologically hydrolyzable carboxyl protecting group, or the pharmaceutically acceptable salts thereof, are potent antibiotics active against various gram-positive and gram-negative bacteria and they may be used, for example, as animal feed additives for promotion of qrowth as preservatives in food, as bactericides in industrial applications, for example in waterbased paint and in the white water of paper mills to inhibit the growth of harmful bacteria and as disinfectants for destroying or inhibiting the growth of harmful bacteria on medical and dental equipment. They are especially useful, however, in the treatment of infectious disease in humans and other animals caused by gram-positive or gram-negative bacteria.

The pharmaceutically active compounds provided by the novel process of this invention may be used alone or formulated as pharmaceutical'compositions comprising, in addition to the active carbapenem ingredient, a pharmaceutically acceptable carrier or diluent. The compounds may be administered by a variety of means; those of principal interest include: orally, topically or parenterally (intravenous or intramuscular injection). The pharmaceutical compositions may be in solid form such as capsules, tablets, powders, etc., or in liquid form such as solutions, suspensions or emulsions. Compositions for injection, the preferred route of delivery, may be prepared in unit cose fern in ampules or in multidose containers and may contain formulatory agents such as suspending, stabilizing and dispersing agents. The compositions may be in ready to use form or in powder form for reconstitution at the time of delivery with a suitable vehicle such as sterile water.

* The dosage to be administered depends to a large extent on the particular compound being used, the particular composition formulated, the route of administration, the nature and condition of the host and the particular situs and organism being treated.

» Selection of the particular preferred dosage and route of application, then, is left to the discretion of the therapist.

In general, however, the compounds may be administered parenterally or orally to mammalian hosts in an amount of from about 5 to 200 mg/kg/day. Administration is generally carried out in divided io doses, e.g. three to four times a day.

The following examples illustrate but do not limit the scope of the present invention.

Example 1 Preparation of 3-(2-(l-pyrldiniuni)ethylthio) -6« -(1-(R) hydroxyethyl ) -7-oxo-l -azablcycloQ.2.0) -hcgt-2ene-2-carboxylate A. 1-(2-mercaptoethyl)pyridlnium methane sulfonate To a suspension of pyridinium methanesulfonate in pyridine prepared by the dropwise addition of methanesulfonic acid (1.95 mL, 0.03 mol) to pyridine (8.0 mL, 0.099 mol) with cooling, was added ethylene sulfide (1.96 mL, 0.033 mol). The resulting mixture was stirred at 55°C for 16 h and concentrated under reduced pressure to a thick syrup which was mixed with few mL of water. The solution was poured on top of a column (40 x 16 cm) of p-bondapak C-18 which was eluted with water. Lyophylization of the appropriate fractions gave a colorless syrup 6.5 g (91a), ir (film)uraax: 2300-2600 (br, SH), 1635 (pyridinium), 1490, 1200 (sulfonate), 1068, 1060, 1045, 791, 780 cm-1, 1Hmr (DMSO-d6> 6: 2.32 (3H, s, CH3SO3-), 2.61,2.70, 2.73, 2.82 (1H, B part of A2B system, SH), 3.07 (211, m [with D2O, 3.08 (2H, t, J=6.5 Hz) ], Cll2S), 4.76 (2H, t, J-6.5 Hz, C112N+), 8.19 (211, m, Rra of pyridinium), 8,6 (1H, a, Ho of pyridinium), 9.08 (2H, dd, J-6.8 Hz, J-1.4 Hz, Ho of pyridinium), uv (H20) Amax: 206 (£5230), 258 (£ 3760) rap. 9 METHOD A B. 1- (l-mercaptoethyljpyridiniuiEi chloride MsO Permutit S-l Cl An aqueous solution of crude 1-(2-mereaptdethyl)pyridinium methanesulfonate (9.4 g, 0.04 mol) was poured on top of column (2.5 x 41 cm) of per&utitT S-l Cl* The column was fluted with water at a rata of 0.5 ni per min and the appropriate fractions were combined and lyophylised giving a yellowish syrup 7.0 g (100%) which was used as it was for the next seep» ~Hsr (0^0) 6: 3.22 (sH* mP CHjS), 4.83 (ms 0¾^ *)s 8·1® (2Β» h; fe of pyridinium), 8.7 (lHj κι, Hp of pyridinium), 9.0 ppm (2HP Ho of pyridiniwsi) .

To a precooled (ies hath) pyridine (5.6 ©LP 70 mmol) was added pyridine hydrochloride (4.05 g, 35 mol) and ethylene sulfide (2.1 mLp 35 mol). The mixture was heated at 65 °C and stirred for 75 min to give a two phases system. The lighter phase was removed. The remaining oil was washed with ether (5 x 10 mi) and pumped under high vacuum to give the title compound (90-100X) which was used as such for th© next step.

Trade Mark C. Paranitrobengyl 3- (2-(lpyridinium)ethylthio )-6gg - [I-(K)-hydroxyethyl 1-7-oxo-l-agabicyclo(3.2,0)hepcene-2-carboxylate chloride 1- NSsU?;) xo , cj ,{OPh) 'COOFNB 3- ^SZV^0) ci 4- NEt(£?r) A solution of p-nitrobensyl 6s«· [1-(&) -hydroxyethyl 3-3,7-dloxo~l-asabicyclo(3-2.0) heptane-2-carboxylete (6.09 g, 17θ5 ©mol) in acetonitrile (20 ®L) cooled to °C under a nitrogen atmoophsre and treated successively with di10 isopropylethylamlne (3.65 mL, 21.0 mmol) and diphenyl chlorophosphate (4.34 mL9 21.0 mmol). The resulting mixture was stirred for 30 min at 5°CP cooled to -5°C and treated successively with a solution of crude l"(2-raerceptopN~diraethyI£or®amide (1.0 n&) and dropwise ( with diisopropylethylamine (3=65 sL, 21.0 mmol). The reaction mixtux® was stirred at 0°C for 1 hr, cooled to ~30°C and stirred for 15 min. more. The solid was filtered off and washed with cold (-30°C) eceeonicrile 5.77 g (65«), it (nujoX) Vmax · 3300 (OH) 8 1775 (C-0 of S-Uccam), 1690 (C-0 of W9B escsr) , 1630 (py-fid ini.ua) , 1605 (phsayl of ?MB ester), 1515 (W02), 1335 cm'1 (HO?), ’-Iter (DHSO-ds) 6: 1.17 (3H, <2, J-6.1 Hsp CH3CHOH), 3.2-3.75 <5H, H-4, Trade Mark Si H-6, CH2S), 3.75-4.5 (ZH, H-5, CH3CHOH), 4.92 (2H, brt, J=6.5 Hz, CH2N+), 5.18 (1H, d, J=4.9 Hz, OH), 5.37 (center of ABq, Hz, CH2 of PNB), 7.69 (2H, d, J-8.7 Hz, Ho of PNB), 8.24 (d, J8.7 Hz, Hm of PNB), 8.0-8.4 (411, Hm of PNB, Hm of pyridinium), δ.66 (1H, m, lip of pyridinium), 9.17 (2H, brd, J = 5.5 Hz, Ho of pyridinium). The filtrate and washing were combined and diluted with ether (150 mL). The supernatant was decanted and the gum was dissolved in water (40 mL) containing enough acetonitrile to have a solution which was poured on top of a column (3 x 10 cm) of μ-bondapak C-18. The column was eluted with 10% acetonitrile - 90% water (150 mL) and 50% acetonitrile 50% water (100 mL) mixtures. The appropriate fractions were combined and lyophylized after the acetonitrile has been removed under vacuum giving a yellowish powder. An NMR of it showed the presence of the title compound mixed with some p-nitrobenzyl 3-[2-(l-pyridinium)ethylthio]-6c-Cl-(R)-hydroxyethyll-7-oxo-l-azabicyclo(3.2.0) hept-2-ene-2-carboxylate diphenylphosphate (2:1). The powder was dissolved in water (minimum amount) and passed through a column (1.5 x 21 cm) of permutit S-1C1~ with water. Lyophy1ization of the appropriate fractions gave 1.8 g (20%) of the title compound.

S3 D. Paranitrobenzyl 3- [2-(l-py_rldj.nium)ethyl_thio ]-&*- [1-(R)-hydroxyethyl ]-7-oxo-l-azabicylo(3.2.Q)kgpt-2°ene-2-carboxylate d i p heny 1 pho .¾ pha te. 1) NEt(iPr)6 2) CU»(0?b)2 -...-.— —__- -> 3) 4) A solution of p-nitrobensyl 6a ° [X"(19-hydroxyethyl 33,7rtdioxo-l~azabicyclo(3.2.0) heptane-2-carboxylate (0.174 g, 0.50 mmol) in acetonitrile (2 aL) vts cooled to 0°C under a nitrogen atmosphere and treated successively with diisopropylethylamine (0.105 mL, 0.60 mmol) and diphenyl chlorophosphate (0.124 mL, 0.60 mmol). The resulting solution was stirred for 30 min at 0°C and treated successively with a solution of l-(2*mercaptoethyl)pyridinium methanesulfonate (0-170 g, 0.72 mmol) in acetonitrile (0.6 mL) and diisopropylethylamine (0.105 mL, 0.60 mmol). The reaction mixture was stirred at 0°C for 15 min, diluted with cold (0°C) water (7 mL) and poured on top of a column (1.5 x 6.4 cm) of ρ-bondapak C-18.

The column was eluted with a mixture of acetonitrile (25%-50%) in water (75%-507,). The appropriate fractions were combined and lyophylized after the acetonitrile has been removed under vacuum giving a yellowish powder 0.33 g (92a), ir (KBr) >raax :3600-3000 (0U), 1765 (OO of β-lactam), 1690 (C*0 of PNB ester), 1625 (pyridinium), 1585 (phenyl), 1510 (N02), 1330 (N02), 885 cnT* (N02), hlmr (DMSO-cf^) ό: 1.16 (3H, d, J-6.2 Hz, CH3CHOH), 4.87 (2Π, brt, 3-6.6 Hz, CH^S), 5.37 (center of ABq, 2a b ~ 14.3 Hz, CHg of PNB), 6.7-7.5 (phenyl), 7.68 (d, 3*8.8 Hz, Ho of PNB), 8.23 (d, J = 8.8 Hz, Hm of PNB), 8.0-8.3 (m, Hm of pyridinium), 8.4-8.8 (1H, Hp of pyridinium), 9,09 (2H, dd, 3=6.7 Hz, 3 = 1.3 Hz, Ho of pyridinium).

E. 3- [2-(1-pyridinium) ethyl thio]Cl-(£j-hydroxyethyl] 7-oxo-l-azabicyclo(3.2,0)hept-2-ene-2-carboxyl.ate Method A ¾ ?<*/C, JL, ..... V TKF, Ether To a solution of p-nitrobenzyl 3-[2-(l-pyridinium) ethyl thio] — 6ελ — [l-(R)-hydroxyethyl] -7-oxo-l-azabicyclo (3.2.0)hept-2-ene-2-carboxylate diphenylphosphate (0.16 g, 0.22 mmol) in wet tetrahydrofuran (10 mL) was added ether (10 mL), potassium phosphate monobasic-sodium hydroxide buffer pH 7.4 (16 mL, 0.05 M) and 10% palladium on charcoal (0.16 g). The resulting mixture was hydrogenated under 40 psi for 1 h at 25°C. The two phases were separated and the organic phase was extracted with water (2x3 mL). The aqueous solutions were combined, washed with ether (2 x 10 r.L) and poured on top of a column (1.5 x 6.2 ern) of ju-bondpak C-18 after the traces of organic solvents have been removed under vacuum. Elution of the column with water gave after lyophylization of the appropriate fractions a yellowish powder 0.062 g (84%), ir (KBr) \J : 3700UlQX 3000 (OH), 1755 (C=0 of $-lactam), 1630 (pyridinium), 1590 cm'1 (carboxylate), XHmr (D20): 1.22 (3H, d, J = 6.4 Hz, CH3CHOH), 2.92 (d, J = 9.1 Hz, H-4), 2.97 (d, J=9.1 Hz, H-4), 3.20 (dd, £=2.5 Hz, J=6.1 Hz, H-6), 3.44 (t, J=6.0 Hz, CH2S), 3.93 (dd, J=9.1 Hz, J=2.5 Hz, H-5), 4.82 (t, J=6.0 Hz, CH2N+), 8.04 (m, Hm of pyridinium), 8.5 (m, Hp of pyridinium), 8.82 (dd, J = 3.2 Hz, £=1.1 Hz, Ho of pyridinium), uv (H-0) X : 259 (£5800), 296 (£7030) ιημ, 4» ul&X t1z = 13.5 H (measured at a concentration of 10" M in a phosphate buffer pH 7.4 at 36.8°C).

To a solution of p-nitrobenzyl 3- [2-(l-pyridinium) ethyl thio] -6cs- [l-(R) -hydroxyethyl] -7-oxo-l-azabicyclo (3.2.0)hept-2-ene-2-carboxylate chloride (5.77 g, 11.4 mmol) in potassium phosphate monobasic-sodium hydroxide buffer (170 mL, 0.2 M, pH 7.22) was added tetrahydrofuran (30 mL), ether (30 mL) and 10% palladium on charcoal (5.7 g). The resulting mixture was hydrogenated at 22°C under 4.0 psi for 1 h and filtered on a Celite** pad. The pad was washed with water (2 x 15 mL). The filtrate and washings were combined and diluted with ether (100 mL) The aqueous phase was separated, washed with ether (3 x 100 mL) and poured on top of a column (4.5 x 20 cm) of μ-bondapak C-ιβ after the organic solvants have been removed under vacuum. Elution of the column with water followed by a mixture of 1% acetonitrile in water gave after lyophylization of the appropriate fractions 2.48 g (65%) of the title compound as a yellowish powder. The analytical data were identical to those reported for the compound prepared in the method A.

Trade Mark 7 Example 2 Preparation of 3-f2-( 1-( 3,5-dj rnethyIpyridinium) ethyl thiol]-6<λ-Cl-(&)-hydroxyethy ll -7-oxo-l-azabicyclo A. 1-(2-mereaptoethyl)-3,5-dlmethylpyridiniutn methanesulfonate To a suspension of 3,5-lutidinium methanesulfonate 10 in 3,5-lutidine prepared by the addition of methanesulfonic acid (0.65 mL, 0.010 mol) to cold 3,5-lutidine (2.51 mb, 0.022 mol) was added ethylene sulfide (0.655 mL, 0.011 mol). The resulting mixture was stirred under a nitrogen atmosphere at 55°C for 24 h, cooled to 23°C 15 and diluted with water (5 mL) and ether (5 mL). The organic layer was separated and the aqueous solution was washed with ether (6x4 mL). The traces of ether were removed under vacuum and the solution was applied on top of a column (2.5 x 6.0 cm) of ^μ-bondapok C-18. The column was eluted with water and lyophilization of the appropriate fractions gave a colourless syrup 2.4 g (91%); ir (film) \J t 2520 (SH), 1628 (pyridinium), mux 1600, 1495, 1325, 1305, 1283, 1200 (sulfonate), 1040, 938, 765, 680 cm-1; 1Hmr (DMSO δ: 2.31 (311, s, CH3SO3"), 2.47 (6H, s, CHg on pyridinium), 2.57, 2.66, 2.69, 2.78 (1H, B part of A^B system, SH), 3.06 (2H, π[with D20 added (2H, t, J=6.5 Hz)], CH2S), 4.65 (2H, t, J=6.5 Hz, CH2N+), 8.34 (1H, s, Hp of pyridinium), 8.79 (2H, s, Ho of pyridinium); uv (H^O) 271 mp. Anal, calcd. for C1OH17NO3S2‘°·5H20: C44.09, H 6.66, N 5.14, S 23.54; found: C 44.26, H 6.49, N 5.17, s 24.18.

B. Paranitrobenzyl 3-Q2-( 1-(3,5-ditne thylpy ridlnium)) e thy 1 thiol -6ji-£l·- (gj -hydroxye thy1}-7-oxo-l-azabi eye1 o (3.2.0)hept-2-ene-2^carboxylate diphenylphosphate ‘ 2 . i?hO>3?o To a cold (0°C) solution of p-nitrobenzyl Cd-(1(R)-hydroxyethy1)-3,7-dioxo-l-azabicyclo(3.2.0)hcptane2-carboxylate (0.523 g, 1.50 mmol) in acetonitrile (6.0 mL) kept under a nitrogen atmosphere was added di iso20 propylethylamine (0.314 mL, 1.8 mmol) followed by se diphenyl chlorophosphate (0.373 mL, 1.8 rrcnol). The reaction mixture was stirred for 30 min and treated with a solution of l-(2-mercaptoethyl)-3,5-dimethyl pyridinium methanesulfonate (0.493 g, 1.87 mmol) in acetonitrile (1.9 uL) followed by diisopropylethylamine (0.314 mL, 1.8 mol). The reaction mixture was stirred at 0°C for 1 hr, diluted with cold (0°C) water (26 mL) and poured on top of a column (7.0 x 3.5 cm) of p ir (KBr) 3700-3100 (OH), 1778 (CO of S-laccam), 1700 (C=0 of PH3 ester), 1635 (pyridinium), 1595 (phenyl)s 1521 (K02), 1333 (H02)» 859 ce'1 (MO2), J-Hmr (BMSO d$) %: 1.16 (3Hr d, J-6.1 Hs, CH3CHOH), 2.43 (s, CH3 on pyridinium), 4.75 (2H, m, CHjR1’), 5.38 (center of ABq, 2a,bwl^·® Hs, CHo of PKB) e 6.6-7.5 (10H, m9 phenyl) s 7.70 (2K, d, J"8./ EsP of FO), 8.0.-8.5 (3H, mp Hp of pyridinium, Hm of FNB), 8.82 (2H9 o, Ho of pyridinium), uv (HjO) Xaax:270 (£11570), 306 ( £ 7343) mp. &nal. calcd. for Cg/H^gWgOi 0SP.H2O: C 58.03, H 5.26, H 5.48, S 4.18; found: C 57.98, H 5.05, H 5.22p S 4.34.

C. 3- [2-( 1-(3,5~dime thy lpyridiniuni) )c thyl t.hjo] -6«[l-(jj) -hydroxyethyl]-7-oxo-l-azabicyclo(3.2,0)hept -2-cne-2-carboxylate To a solution of p-nitrobenzyl 3-[2-(1-(3,5dimethylpyridinium)) ethylthio]-6a-[l-(R)-hydroxyethyl] 7-oxo-l-azabicyclo(3.2.0)hept-2-ene-2-carboxylate diphenylphosphate (0.600 g, 0.80 mmol) in wet tetrahydrofuran (36 mL) was added ether (36 mL), potassium phosphate monobasic-sodium hydroxide buffer (0.05M, pH 7.4, 44 mL) and 10% palladium on charcoal (0.60 g). The resulting mixture was hydrogenated under 40 psi at 23°C for 1.25 h. The organic layer was separated and extracted with buffer (2x5 mL). Water layers were combined, filtered through a Celite pad, washed with ether (40 mL), pumped to eliminate traces of organic solvents and poured on top of a column (2.5 x 10.0 cm) of jj-bondapak C-18.

Elution of the column with water and lyophilization of the appropriate fractions gave the title compound 0,186 g (64%) as a yellowish powder, ir (KBr) V : 3700-3100 (OH), 1760 (C=0 of p-lactam), 1595 ern (carboxylate) , blmr (DjO) δ: 1.21 (3H, d, £--6.3 Hz, CH^CHOH), 2.45 (6H, s, CH3 on pyridinium), 2.81 (d, £=9.2 Hz, H-4), 2.96 (d, £=9.2 Hz, H-4), 3.22 (dd, £=2.6 Hz, £=6.2 Hz, H-6), 3.40 (t, £=6.2 Hz, CHgS), 3.84 (dd, £=9.2 Hz, £=2.6 Hz, H-5), 4.15 (m, CH3CH0H), 4.71 (t, J=6.2 Hz, Ciyi*) ,8.21(1H, s, Hp of pyridinUT.) ,8.46(2H, s, Ho of pyridinium), uv (Ho0)^ : 279 (£.8345), 296 λλ 4 max (£7714) m, Mn + 40·7 Example 3 Preparation of (5fi, 6£)-3-[Q2-hydroxymethylpyridlnio) ethyl]thio]-6-ri-(R)-hydroxyethyl]-7-oxo-l-azabicyc]o 13,2.01 hept-2-ene-2-carboxylate CK^CE A. 3-Hyd roxymethyl-1- (2-mercaptoethyl)gyridlnluni trifluoromcthanesulfonate Trifluoroeethanesulfonic acid (1.327 mL, 0.015 mol) was added dropwise to 3-pyridinemechanol (2.91 mL, 0.030 mol), followed by ethylene sulfide (0.89 mL, 0.015 mol). The resulting homogeneous mixture was heated (oil bath) at 50-70°C under N2 for 20 h. The reaction mixture was then taken up in H2O (15 mL) and extracted with CH2C12 (5x5 mL). The aqueous phase was concentrated in vacuo and then applied to a Cyg reverse-phase column. Elution with H2O followed by evaporation of the relevant fractions gave a pale yellow oil. This material was rechromatographed to give a nearly colourless oil. After drying in vacuo (P2°5) this afforded the product (4.50 g, 94%) as a viscous oil. ir (film) Vmax:3450 (s, OH), 2560 (w, SH) cm~l. 1-Hmr (d$-acetone) 6: 9.10-8.05 (m, 4H, aromatic), 5.01 (t, 3"5.5 Hz, 2H, N-CH2), 4.93 (s, 2H, -CH2OH), 4.43 (br S, 1H, -OH), 3.43-3.18 (m, 2H, S-CH2), 2.34-2.10 (m, 1H, SH). 7^ B. p^Hitrobensyl (5R.6S)~3- [2-(3-hydroxymeehylpyrldlnio)sthyl thio 1·6· [l-*(.R)*hydroxyethyl)-7M oxo^l^agablcyclo [3.2.0 ] hept-2-ene 2-carboxylate To ¢4 solution of p-nitrobengyl (5R,6S)-6-[l-(U)hydroxyethyll^DT-dioxo-l-esabicycXo [3.2.0)heptane-2carboxyl&ce (0,1/4 g, 0.50 mmol) in 2 ©L of dry acetonitrile wag added dilsop^opylethylaisin^ (0θ09ο ffils 0.55 nmol) at 0°C und^r Ko. Diphenyl chlorophosphate (0.114 ε!γ 0.55 mmol was then added dropwise find W® reaction mixture wbs stlrrsd at O^C for 30 ©in. A solution S-hydroEymethyl^l-^-aercapcoethyl) pyridinium erifluoromethanesulfonat© (0.223 g, 0.70 mmol) in 0.50 m.t of acetonitrile wag efosn added „ followed by diisopropylochylamine (0.122 eil, 0.70 mmol). After being kept at 0°C for 30 min the reaction mixture was concentrated in vacuo and the residual yellow gum wag taken up in HoO (enough acetonitrile wag added to aid in dissolving the gum). This solution wag applied co a C^g reverse-phas© column which vas aluted with 15t aceeonitrlle-H^O· Lyophilisation of the relevant fractions afforded the product (0.305 g, 8lX) na a beige-coloured solid, ir (KBr) Pmax: 3420 (br, OH), 1775 (β-lactara CO), 1695 (-CO2PNB) cnT**, ^Hmr (dfcacctone) 6: 9.44-7.72 (m, 8H, aromatic), 7.22-6.91 (m, 10H, diphenylphosphate),5.53, 5-27 (ABq,J« 14lIz,2H, benzylic), 5.04 (c, J"7.4 Hs, 2H, H-CH2), 4.75 (s, 2H, CH2OH), 4.5-3.1 (m, 8H), 1.21 (d,J6.3 Hz, 3H, CHHe).

C. (5Rt6S)-3- [2-(3-hydroxymethylpyridinto)ethyl) ehlo 3-6- [l-(R)-hydroxyet:hyl)-7-oxo-l-azabicyclo [3.2.0 ]hept-2-ene-2-carboxylate r, To a solution of p-nitrobenzyl (5R,6S)-3- [2(3-hydroxymethylpyridinio)ethyl thio ]-6- [1-(R)hydroxyethyl ]-7-oxo-l-azabicyclo [3.2.0 ]-hept-2ene-2-carboxylate diphenylphosphate (0.145 g, 0.194 mmol) in 10 mL of THF containing 5 drops of Η2 0.145 g of 10% palladium-on-charcoal and 10 mL of ether. The mixture was hydrogenated (Parr) at 40 psi for 1 h and then filtered through a pad of Celite.

The filter cake was washed with a little H2O and ether and the aqueous phase was separated and extracted with ether (3x). The aqueous solution was then cooled at 0°C and the pH was adjusted to 7.0 with pH 7.4 buffer. After removing residual volatiles in vacuo the aqueous solution was applied to a C^g reverse-phase column which was eluted with H2O. Lyophilization of the relevant fractions gave Che product (36 mg, 51%) as a light yellow solid. Further purification by reverse-phase hplc gave the pure product (31 mg, 41%) as a solid. ir (KBr) 7? »max: 33OO(br, OH), 1755 ( fl-lactam CO), 1590 (-C02) cm’I; ^Hnmr (DjO) 6: 8-78-7-94 (m,4tl aromatic), 4.83 (t, >6.0 Hz, 2H, W-CH2), 4.83 (s, 2H, CH2OH), 4.16 (d of q, J»J’-6.2 Hz, 1H, H-l'), 3.98 (d, of t J-9.1 Hz, J’2.6 Hz, IH, H-5), 3.75-3.20 (m, 3H), 3.20-2.65 (m, 211), 1.22 (d, >6.4 Hz, 3H, CHMc); uv (H2O) W 294 (£ 7614), 266 (£6936) nm; t(/ (pH 7.4 36.8°C) 14.0 h.

Example 4 Preparation of (5R,6S)-3- [2-(4-hydroxymethylpyrldlnlo)ethylthio][l-(R)-hydroxyethyl ]-7-oxo-l-azabicyclo [3.2.Ojhep 0 2-ene-2-carboxylate 7$ A. 4-Hyd roxym*? thy 1-1-(2-mereap toethyl )pyrldinlum trifluoromethanesulfonate CS^CE αΓ3503Η "Wifi To a solution of 4-pyridinemethanol (1.635 g, 0.015 mol) in 10 mL of CH^Cl^t at 0°C under was added dropwise trifluoromethanesulfonic acid (1.327 mL, 0.015 mol).

A yellow-brown oil rapidly separated out. An additional equivalent of 4-pyridinemethanol (1.635 g, 0.015 mol) was added to this mixture and the solvent was removed (0 under reduced pressure to give an oil. To this oil was added ethylene sulfide (0.891 mL, 0.015 mol) and the resulting homogeneous mixture was heated (oil bath) at about 60°C for 3 h. The reaction mixture was then taken up in 15 mL of H20 and the aqueous solution was washed with CH^Cl^ (5x5 mL). After removing residual organic solvent in vacuo the aqueous solution was applied to a C1Q reverse-phase column. Elution with HgO and subsequent evaporation of the relevant fractions afforded an oil which was further dried in vacuo over PgO^ to 2ive product (4.64 g, 97%) as a colourless oil. ir (film) V : 3455 (s, OH), 2565, (w, SH) cm ^Hnmr (4c-acetone) max o S: 9.07, 8.18 (ABq, J=6.8 Hz, 4H, aromatic), 5.03 (s, 2H, CH2OH), 4.96 (t, 3 = 6.5 Hz, 2H, N-CH2), 4.09 (br s, 1H, -OH) 3.5-3.1 (m, 2H, S-Cflg), 2.25 (brs; 1H, -SH). so Β. p-Nl t robr nzy 1 ( 5£ , GS)-3- [2- ( 4-hydroxyme thylpyridinio) ethyl thio] -6- [l-(R)-hydroxyethyl] -7-oxo-l-azitbicyclo [3.2.0'l h c p t - 2 - e n e - j? - c a rbo x y 1 a t e dip he n y 1 p ho s p h a t e .15 To a solution of p-nitrobenzyl (5R,6S)-6-[l-(R)hydroxyethyl]-3,7-dioxo-l-azabicyclo[3.2.0]heptane-2carboxylate (0.348 g, 1.0 mmol) in 6 mL of dry acetonitrile, at 0°C under N2, was added dropwise diisopropylethylamine (0.191 mL, 1.1 mmol) followed by diphenyl chlorophosphate (0.228 mL, 1.1 mmol). The resulting golden-yellow solution was stirred at 0°C for 40 min.

To this solution was added a solution of 4-hydroxymcthyl1-(2-mereaptoethy1)pyridinium trifluoromethanesulfonate (0.447 g, 1.4 mmol) in 1 mL of acetonitrile, followed by diisopropylethylamine (0.191 mL, 1.1 mmol). A reddishblack gum separated from the reaction mixture. After 20 min at 0°C the reaction mixture was filtered and concentrated in vacuo. The residue was taken up in a minimum volume of acetonitrile-F^O (1:1) and applied to a Clg reverse-phase column. Elution with ?5% acetonitrile Η2θ emd subsequent lyophilization of the relevant fraction gave the product (0.353 g, 47%) as a cream-coloured solid. ir (KBr)V :3240 (br, OH), 1775 (β-lactam CO) max . r 1695 (-COgPNB) cm'1; lHnmr (dg-acetone) 6: 9.24-7.84 (m, 8H, aromatic), 7.4-6.9 (m, 10H, diphenylphosphate), .52, S.24 (ABq, J = 14 Ha, 2H, benzylic), 5.154.80 (m, 4H), 4.45-3.05 (m, 7H), 1.35 (d, J=6.6 Hz, 3H, CHMe).

C . (5&, 6^)-3-Cg-(4-hyHroxymethylpyrid inio)ethyl thio]6- [l-( H ) -hydroxys thyl] -7-oxo-l-azabicyclo [3. 2 .0] hopt-2-one-2-carboxyIate A mixture of p-nitrobenzyl (5R,6S)-3-Q2-(4-hydroxymethylpyridinio)ethyl thio]-6- [l-(R)-hydroxyethyl]-7-oxol-azabicyclo [3.2.0]hept-2-ene-2-carboxylate diphenylphosphate (0.348 g, 0.465 mmol) and 10% palladium-oncharcoal (0.35 g) in 11 mL of phosphate buffer (0.05 M, pH 7.4), 5 mL of THF and 10 mL of ether was hydrogenated at 40 psi for 1.25 h. The mixture was then filtered through a Celite pad and the aqueous phase was washed with ether (3 x). The pH of the aqueous solution was then adjusted to 7.0 using additional pH 7.4 buffer.

After removing residual volatiles in vacuo the aqueous solution was applied to a C1Q reverse-phase column. Elution with 2% acetonitrile-HgO and subsequent lyophilization gave a yellow-brown solid. This material was rechromatographed (C^q reverse-phase/I^O) to give the desired product (0.060 g, 36%) as a light yellow solid. ir (KBr)^ · 3400 (br, OH), 1755 (ί-lactam CO). 1590 ~ max (-C02 ) cin-1; innmr (DgO) 6: 8.73, 7.96 (ABq, J-6.8 Hz, 4H, aromatic), 4.93 (s, 2H, CHgOH), 4.77 (t. J-6.0 Hz, 211, N-CH.,), 4.15 (d of q, J=J' = 6.3 Hz, 1H, H-l ·), 3.96 (d of t, J=9.2 Hz, J'=2.6 Hz, 1H, H-5), 3.65-3.20 (m, 3H) 3.13-2.62 (m, 2H) , 1.21 (d, J=6.3 Hz, 3H, CHMe); uv (1-^0) : 295 (£6880), 256 (£S595), 224 (£8111) nm; tk max 'ιζ (pH 7.4, 36.8*C) 14.5 h Example 5 Preparation of 3- [2-( l-ί 2-me thylpyridinium))ethylthio]-6«-[l-(R)-hydroxyethyl]-7-oxo-lazabicyclo(3.2.0?hept-2-ene-carboxyla to A. 1-(2-mercaptoethyl)-2-methylpyridinium methanesulfonate To a suspension of 2-methylpyridinium methanesulfonate in 2-methylpyridine prepared by the addition of methanesulfonic acid (0,65 mL, 0.010 mol) to cold 2-methylpyridine (2.17 mb, 0.022 mol) was added ethylene sulfide (0.655 mL, 0.011 mol). The reaction mixture was stirred under a nitrogen atmosphere at 55°C for 21 h, cooled to 23°C and diluted with water (5 mL). The aqeuous solution' was washed with ether (6x4 mL) pumped to remove traces of organic solvents and poured on top of a column (2.5 x 10.0 cm) of ρ,-bondapuk C-18. The column was eluted with water and lyophilization of the appropriate fractions gave 2.13 g (85%) of the title compound, ir (film) -J__: 2520 (SH), max 1623 (pyridinium) 1574, 1512, 1485, 1412, 1195 (sulfonate), 1038 cm1, 1Hmr (DMSO-rfg + DgO) δ: 2.37 (3H, s, CHgSOj), 2.83 (3H, s, CH3 on pyridinium)., 3.09 (2H, J=6.9 Hz, CH2S), 4.71 (2H, t, J=6.9 Hz.CHgN*), 7.93 (2H, m, Hm of pyridinium), 8.44 (1H, tn, Hp of pyridinium), 8.89 (1H, m, Ho of pyridinium), uv (H20) Xmax: 266 (£3550) mp.

B. Paranitrobenzyl S-fe-Cl-^-methylpyridinium) ethylthio] [l-(K)-hydroxyethyl] -7-oxo-l-azabicyclo(3.2.0) hept-2-ene-2-carboxylate diphenylphosphate To a cold (0°C) solution of p-nitrobenzyl 6*-[l-(R)15 hydroxyethyl]-3,7-dioxo-l-azabicyclo(3.2.0)heptane-2carboxylate (0.523 g, 1.50 mmol) in acetonitrile (6 mL) kept under a nitrogen atmosphere was added diisopropylethylamine (0.314 mL, 1.80 mmol) followed by diphenyl chlorophosphate (0.373 mL, 1.80 mmol). The reaction mixture was stirred for 30 min at 0QC and treated with solution of 1-(2-mereaptoe thy1)-2-methylpyridinium methanesulfonate (0.530 g, 2.16 mmol) in acetonitrile (18 mL) followed by di isopropylethylamine (0.314 mL, 9C .10 1.8 mmol). The reaction mixture was stirred at 0°C for 1 h diluted with cold (O°C) water (26 mL) and poured on top of a column (3.5 x 7.0 cm) of ^Λ-bondapak C-18.

Elution of the column with 25% acetonitrile - 75% water and with 50% acetonitrile - 50% water gave after lyophilization of the appropriate fractions 1.06 g, (96%) of the title compound as a yellowish powder, ir (KBr) : 3650-3100 (OH), 1770 (C=0 of P-lactam), 1695 and 1690 (C=0 of PNB ester), 1630 (pyridinium), 1595 (phenyl), 1518 (N02), 1335 (NOj), 890cm-1 (N02), 1Hmr (DMSO, S: 1.15 (3H, d, J = 6.1 Hz, CH^CHOH), 2.87 (s, CHg on pyridinium), 3.6-4.4 (2H, m, H-5, CH3CH0H), 4.75 (2H, m, CHgN*), 5.37 (center of ABq, J = 14 Hz, CH,, of PNB), 6.5-7.4 (10 H, m, phenyl), 7.70 (2H, d, J=8.8 Hz, Ho of PNB), 8.0 (2H, tn, Hm of pyridinium), 8.24 (2H, d, 3=8.8 Hz, Hm of PNB), 8.50 (1H, m, Hp of pyridinium), 8.95 (1H, brd, 3 = 6.1 Hz, Ho of pyridinium), uv (Ho0) X : 265 (£11990), 314 (fe8020)mp C. 3-[2-(1-(2-methylpyridinium)) ethyl thio]-6d-[i-(R)hydroxyethyl]-7-oxo-l-azabicyclo(3.2.0)hept-2-enecarboxylate coo To a solution of p-ηίtrobenzyl 3-[2-(l-(2-methylpyridinium))ethyILhio)-6 ok-[l-(R)-hydroxyethyQ7-oxo-l-azabicyclo(3.2,0)hept-2-ene-2-carboxylate diphenylphosphate (0.66 g, 0.90 mmol) in wet tetrahydrofuran (34 mL) was added ether (34 mL), potassium phosphate monobasic-sodium hydroxide buffer (0.15 M, 16.5 mL. pH 7.22) and 10% palladium on charcoal (0.66 g). The resulting mixture was hydrogenated under 40 psi at 23eC for 1.25 h. The organic layer was separated and extracted with buffer (2x6 mL). Water layers were combined, filtered through a Celite pad, washed with ether (40 mL), pumped to eliminate traces of organic solvents and poured on top of a column (2.5 x 10 cm) of ^s-bondapak C-18. Elution of the column with water and lyophylization of the appropriate fractions gave the title compound 0.098 g (31%) as a yellowish powder, ir (KBr) V a : 3650-3100 (OH); 1755 (0=:0 of /J-lactam), 1630 (pyridinium) , 1595 cm' (carboxylate), 1Hmr (D^O) δ: 1.20 (3H, d, £=6.3 Hz, CH3CHOH), 2.83 (s, CH3 on pyridinium), 2.7-3.1 (5H, H-4, CH3 on pyridinium), 3.1-3.7 (3H, m, CH2S, H-6), 3.90 (dd, £=9.1 Hz, £=2.6 Hz, H-5), 3.1 (m, CH3OHOH), 4.78 (t, £-6.2 Hz, CHgN*), 7.8 (2H, m, Hm of pyridinium), 8.3 (1H, m, Hp of pyi'idinium), 8.65 (1H, m, Ho of pyridinium), uv (h2°) λ max:268 (£-9350), 296 (&8840) mji, [<]β23 +41® (c 0.5, HgO), = 15.0 h (measured at a concentration of 104 M in phosphate buffer pH 7.4 at 36.8®C).

Example β Preparation of 3-Γ,2-( l-(4-fnethylpyrjdtnium)cthy 1 thio] -691- Dili J-hydroxye thyl] -7-οχο-ΐazabicyclo(3.2.0)hept-2-ene-2-carboxylate A. 1-(2-mereaptoethyl)-4-methylpyridinium methanesulfonate To a suspension of 4-picolinium methanesulfonate in 4-picoline prepared by the addition of methanesulfor.ie acid (0.65 mt, 0.010 mol) to 4-picoline (2.14 mL, 0.022 mol·) in cooling was added ethylene sulfide (0.655 mL, 0.011 mol). The reaction mixture was stirred under a nitrogen atmosphere at 55°C for 24 h, cooled to 23°C and diluted with water (5 mL) and ether (10 mL). The organic layer was separated and the aqueous layer was washed with ether (5x5 mL) and applied on top of a column (2.5 x 10 cm) of jA-bondapak C-18 after traces of ether have been removed under reduced pressure. Elution of the column with 15% acetonitrile 85% water mixture gave after lyophylization of the appropriate fractions a colorless syrup 2.66 g (100%), ir (film) Vmax: 2500 (SH), 1640 (pyridinium), 1572, 1520, 1478, 1200 (sulfonate), 1040, 833 and 768 cm1, XHmr (DMSO-βθ) <3: 2.31 (3H, s, CH3SO3"), 2.62 (a, CH3 on pyridinium), 2.2-2.9 (4H, SH, CH3 on pyridinium), 3.04 (2H, m, CHgS), 4.68 (2H, t, J=6.4 Hz, CH2iT), 8.01 (2H, d, J=6.6 Hs, Hm of pyridinium), 8.89 (2H, d, J=6.6 Hs, Ho of pyridinium), uv (Ho0) 1 „ : 256 (¢4100), 221 («7544) mu.

B. 1-(2-mercaptoethyl)-4-mgthylpyridinturn p-toluene10 sulfonate To a suspension of p-toluenesulfonic acid (1.72 g, 0.01 mol) in benzene (6.5 mL) was added 4-picoline (1.17 mL, 0-0X2 ffiol). The resulting mixture was stirred under a nitrogen atmosphere at 23eC for 30 min, treated with ethylenesulfide (0.65 mL, 0.011 mol) and stirred at 75° for 24 h, More ethylenesulfide (0.65 mL, 0.011 mol) was added and the stirring was continued at 75°C for 24 h more. The reaction mixture was cooled to 23°C and diluted with water (S mL) and ether (8 mL). The aqueous layer was separated and washed with ether (3x8 mL). The traces of organic solvents were removed under vacuum and the compound was chromatographed on ^a-bondapak C-18 with water as eluting solvent to give 2.94 g (90%) of the title compound as a colorless syrup; ir (film : 2510 (SH) 1640 (pyridinium), max 1595, 1582, 1475, 1200 (sulfonate), 1031, 1010, 818 cm’1, ^’Hmr (DMSO.dg) δ: 2.29 (3H, s, CH3 on pyridinium), 2.61 (s, CH3 Ph), 2.4-2,8 (4H, SH, CH3 Ph), 3.03 (2H, mCaddition of DgO gave a t, £= 6.4 Hz, at 3.04], CHjS), 4.68 (2H, t, £=6.4 Hz, ΟΗ^'Γ) , 7.11,,7.49 (4H, 2d, £=7.9 Hz, Phenyl), 8.00 (2H, d, £=6.5 Hz, gm of pyridinium), 8.89 (2H,d,J-6.5 Hz, Ho of pyridinium), uv (H-0) X _ 256 (£4315), 222 (£17045) mp. d max r C. Paranitrobenzyl 3-[5-(l-(4-methylpyridinium) )ethylΐ;ίΐίάί-6β!-4ΐϋ5ί)-hydroxyethyl] -7-oxo-l-azabicyclo(3.2.0) hept-2-ene-2-carboxylate diphenylphosphate To a cold (OeC) solution of p-nitrobenzyl (R)-hydroxyethyl]-3,7-dioxo-l-azabicyclo(3.2.0)heptane2-carboxylate (0.522 g, 1.5 mmol) in acetonitrile (6 mL) kept under a nitrogen atmosphere was added diisopropylethylamine (0.314 mL, 1.8 mmol) followed by diphenyl chlorophosphate (0.373 mL, 1.9 mmol). The reaction mixture was stirred for 45 min and treated dropwise with a solution of l-(2-mercaptoethyl)-4-methylpyridinium methanesulfonate (0.539 g,2.16 mmol) in acetonitrile (1.8 mL) followed by diisopropylethylamine (0.314 mL, 1.8 mmol). The reaction mixture was stirred at 0°C for 1 h, diluted with cold (0°C) water (24 mL) and poured on top of a column (2.5 χ 8.5 cm) of μ-bondapak C-18. Elution of the column first with 25% acetonitrile -75% water mixture (100 mL) then with 50% acetonitrile 50% water mixture (100 mL) afforded after lyophylization of the appropriate fractions 0.91 g (83%) of the title compound as a yellowish powder, ir (KBr) 3700-2800 (OH), 1770 (C=0 of/3-iactam), 1700 (C=0 of PNB ester), 1640 (pyridinium), 1595 (phenyl), 1520 (N02), 1340 (NO.,), 890 cm-1 (N02), 1Hmr (DMSO.Jg) fi: 1.16 (3H, d, J=6.2 Hz, CH3CH0H), 2.61 (s, CH3 on pyridinium), 3.1-3.7 (3H, m, H-6, CH2S), 3.7-4.4 (2H,m,H-5, CH3CHOH), 4.79 (2H, brt, J=6.3 Hz, CH2N*), 5.17 (d, J=4.9 Hz, OH), 5.37 (center of ABq, J=14.1 Hz, CH2 of PNB), 6.7-7.4 (10 H, m, phenyl), 7.69 (2H, d, J=8.8 Hz, Ho of PNB), 8.00 (2H, d, J=6.5 Hz, Hm of pyridinium), 8.23 (2H, d, J=8.8 Hz, Hm Of PNB), 8.92 (2H, d, J=6.5 Hz, Ho of pyridinium), uv (H„0) λ : 262 (£10835), 311 fe9670) mp. Anal, calcd. for C36H36N3°1OSP'1-5 H20: C 56.84, H 5.17, N 5.52, S 4.21; found C56.89, H 5.13, N 5.19, S 4.41.

D. 3-f2-( i-( 4-methylpyridinium) ethyl thio] -6JX- [?!-(&)hydroxy ethyl) -7-oxp-l-azablcyclo(3.2.0)hept-2-ene-2-Cc;rboxylate CH ¾ ?£/C, 52 TXT, ether, buffer * C To a solution of p-nitrobenzyl 3-(5-(1-(4mcthylpyridinium)) -c thy 1 thio] -6 A- (1-( R)-hydroxye thyl)7-oxo-l-azabicyclo(3.2.0)hept-2-ene-2-carboxylate diphenylphosphate (0.507 g, 0.80 mmol) in wet tetrahydrofuran (30 mL) was added ether (30 mL), potassium phosphate mono basic-sodium hydroxide buffer (0.15 M, 14.7 mL, pH 7.22) and 10% palladium on charcoal (0.59 g) The resulting mixture was hydrogenated under 40 psi at 23°C for 1.25 h. The organic layer was separated and extracted with the buffer. (2x6 mL). The aqueous extracts were combined, filtered through a Celite pad, washed with ether (3 x 20 mL), pumped to remove traces of organic solvents and poured on top of a column (2.5 x 10 cm) of ji-bondapak C-18. Elution of the column with water and lyophylization of the appropriate fractions gave 0.136 g (49%) of the title compound as a yellowish powder-, ir (KBr) V m : 3700-3000 (OH), 1770 (C=0 of /3-lactam), 1642 (pyridinium) 1592 cm (carboxylate), 1Hmr (D20) δ: 1.19 (3H, t, J=6.3 Hz, CH3CH0H), 2.59 (3H, s, CH^ on pyridinium), 2.84 (d, J=9.1 Hz, H-4), 2.90 (d, J=9.1 Hz, H-4), 3.0-3.6 ( 3H , m, CH2S, H-6), 3.86 (dd, J=9,l Hz, J=2.6 Hz, H-5), 4.12 (m, CH3CHOH)), 4.5-4.9 (CH2N+ masked by HOD), 7.80 (2H, d, J=6.6 Hz, Hm of pyridinium), 8.58 (2H, d, J-6.6 Hz, Ho of pyridinium), uv (Ho0) A : 256 (£5510), 262 (e53G0) max 296 (£7050) m, Ma + 20.8° (C 0.48, ^0), T^=12.8 h (measured at a concentration of 10"4 M in a phosphate buffer pH 7.4 at 36.8°C).

S3 Example 7 Preparation of (5¾) 3- t(4-methylthiopyrldinio)ethyl thio ] -(65) - ( (IR) -hydroxyethyl 1 - 7ipxo-l_azablcyclo [ 3.2.0 ]hept-2-ene-2-carboxylate 4-Mercaptopyrldine (5.55 g, 50.0 mmol; Aldrich) was dissolved in boiling abs. EtOH(50 mL). The insoluble material was removed by filtration over Celite. The filtrate was heated to re-dissolve, and when it cooled to ca. 50°C, methyl iodide (3.17 mL, 51.0 mmol; Aldrich) was added at once. The mixture was cooled to crystallize. Filtration of the solid gave 6.77 g (26.7 mmol, y-53.5%) of the title compound as hydriodide: ^Hmr (D2O) δ: 2.70 (3H, s, "SCH3) and 7.65-7.77-8.35-8.48 ppm (4H, A2B2 type, aromatic Hs); ir (Nujol) yraax: 1615, 1585 (aromatic and 780 cml; uv (H2O) \aX: 227 (£2.02 x 1θζ») and 298 nm (£1.64 x 104).

The hydriodide (6.33 g, 25.0 mmol) was dissolved in Η2θ (40 mL) and the insoluble material was removed and washed with H2O (10 ml). To the filtrate was added at 0-5° NaOH pellet (5 g) and extracted with Et20 (3 x 25 mL), saturating the aqueous layer with NaCl. The combined organic extracts were washed with brine (x 2), dried (MgSG&) and evaporated, yielding 2.92 g (23.4 mmol, overall yield 50%) of the title compound as an oil: ^Hmr (CDCI3) 5: 2.48 (3H, s, -SCH3) and 7.03-7.13-8.38-8.48 ppm (4H, A2B2 type, aromatic-Hs); ir (film)ymax: 1580 and 800 cm’l. ^Preparation of this compound was reported by King and Ware, J. Chem. Soc., 873 (1939). The procedure, described in this reference was followed.

B. 4-Mathylthio-N-(2-mercaptoethy1)pyridinium methanesulfonate 4-Methylthiopyridine (2.75 g, 22.0 mmol) was 20 added slowly to methanesulfonic acid* (0.65 mL, 10.5 mmol) by cooling in an ice-bath. To this solid was added ethylene sulfide* (0.66 mL, 11.0 mmol, Aldrich) and the mixture was heated at 50-60°C for 21 h. As reaction proceeds the solid went to solution. After cooling, the reaction mixture was dissolved in H2O (5 mL) and washed with Et20 (5x4 mL). The cloudy aqueous layer was filtered over Celite and the filtrate was purified by reverse phase silica gel column 5 chromatography (C^g micro bondapack 10 g) eluting with H2O. Each fraction of 10 mL was collected. Fractions 2 and 3 were combined and repurified by the reverse phase column. Fraction 2 gave 1.258 g (4.48 mmol, y. 42.6a) of the title compound as a viscous oil: 1Hmr (DMSO-da, CFT-20) δ: 2.32 (3H, s, MeSo3 G), 2.72 (3H, s, -SMe), 2.68 (1H, m, SH), 2.9-3.2 (3H, ra, -CH2S-), 4.59 (2KP t, J-6.4 Hz, -CH2N©), 7.97 (2H, d, J«7.2 Hz, aroaatic-Hs) and 8.72 ppm (2H, d, Jw7-2 Hz, aromatiC"Hs); ir (neat) Vmax: 1630, 1200 (br, -SO3<5), 7.83 and 770 cm1.

*These reagents were· distilled prior to use.

C. (5&) p-Nitrobenzyl 3- [2~(4-Methylthiopyridino) ethylthio )-(65)- [(lK)-hydroxyethyl ]-7-oxo-lazabicyclo [3.2.0 ]hept-2-ene-2-carboxylate chloride To a solution of (5R) p-nitrobenzyl 3,7-dioxo(6S)- [(IR)-hydroxyethyl ]-l-azabtcyclo [3.2.0 ] heptane-(2R)-carboxylate (475 mg, 1.36 mmol) and diisopropylethylamine (0.24 mL, 1.4 mmol) in CH3CN (5 mL) was added at 0°-5°C under a nitrogen a-tmosphere diphenyl chloro* 9G phosphate (0.29 mL, 1.41 mmol). The mixture was stirred at 0°-5°, for 30 min. To this mixture was added an oily suspension of 4-roethyithio-N-(2mercaptoethyl)pyridinium methanesulfonate (678 mg, 1.45 mmol; 60% pure) In CH3CN (1.5 mL) followed by diisopropylethylamine (0.24 raL, 1.4 mmol). The mixture was stirred at 0°"5°C for 1 h. Immediately after addition of the base, yellowish precipitate formed. The precipitate was filtered and washed with cold CH3CN (3 mL), yielding 413 mg of yellowish solid. This was triturated from 10% MeOH In H2O (5 mL) to obtain 34l mg (0.618 mmol, y. 45-4%) of the title compound as white crystals: mp 118°-120°C; iHmr (DMSO-de, CFT-20) S: 1.16 (3H, d, >6.1 Hz, l’-CH3), 2.72 (3H, s, -SCH3), 3.1-3.7 (5H, m^, 3.7-4.3 (2H, m), 4.71 (2H, t, >6.3 Hz, -CH2N), 5.15 (1H, d, >4*9 Hz, OH), 5.2Ο-5.35-5.4Ο-5.55 (2H, ABq, CO2CH2-Ar), 7.70 (2H, d >8.8 Hs, nitrophenyl-Hs), 7.97 (2H, d, >7.0 Hz, pyridinio-Hs), 8.25 (2H, d, >8.8 Hz, nitrophenyl-Hs), and 8.76 ppm (2H, d, >7.1 Hz, pyridinio’Hs); ir (Nujol)ymax: 3250 (OH), 1775 (B-lactam), 1700 (ester) and 1625 cml (pyridinio); uv (abs, EtOH) Araax: 308 nm (£4.47 x 104); [«]o3° + 24.8 (c 0.5, MeOH); Anal. calcd. for C24H26N3O6S2C1.H2O: C 50.56, H 4.95, N 7.37; found: C 50.63, H 4.72, N 6.89.

D. (5R) 3- [?-(4-Hethylthiopyridinio)ethylthio) - (6£) [(IB)-hydroxyethyl)-7-oxo-l-azablcyclo [3.2.0)hept -2-ene-2-carboxylate OH H2/?d-C -SMe pH 7.4 buffer C°2?XB SMe (5R_) p-Nitrobenzyl 3- ^2-(4-methy1thiopyridinio) e thylth i oj - ( 6S) - [{IR)-hydroxyc thyl]-7-oxo-l-azabicyclo [3.2.0]hept-2-ene-2-carboxylate chloride (380 mg, 0.688 mmol) was dissolved in THF (31.5 mL) and pH 7.40 phosphate buffer (31.5 mL; 0.05M Fisher) and diluted with EtgO (31.5 mL). This solution was mixed with 10% Pd-C (380 mg, Engelhard) and hydrogenated at 35 psi on the Parr shaker at room temperature for 1 h. The aqueous layer was filtered over Celite to remove the catalyst and the Celite pad was washed with H£0 (2x5 mL). The filtrate and washing were combined and washed with Et^O (2 x 30 mL).

The aqueous layer was pumped off to remove any organic solvents and purified by reverse phase column chromatography (C18 microbondapak, 13 g, Waters Associates) eluting with H^O. Fractions having a uv absorption at 307 nm were collected (ca. 1 L) and lyophilized to obtain 127 mg (0.334 mmol, y. 48.5%) of the title compound as a yellowish powder: 1Hmr (DgO, CFT-20) 6: 1.20 (3H, d, £=6.4 Hz, 1' -CH3), 2.64 (3H, s, -SCHg), 2.81 (2H, m, -SCH2-), 3.19 (1H, dd, £6_lt =6.1 Hz, £θ_5=2.6 Hz, 6-H), 3.32 (2H, dd, £=11 Hz, £=5.5 Hz, 4-Hs), 3.92 (1H, dt, J = 9.2 Hz, £56=2.6 Hz, 5-H), 4.1 (1H, m, 1' -H), 4.61 (2H, t, £=5.9 Hz, -CH2N®), 7.70 (2H, d, £=7.1 Hz, aromatic-Hs), and 8.40 ppm (2H, rf, J = 7.1 Hz, aromatic-Hs) 25 ir (KBr, disc)V : 3400 (OH), 1750 (tf-lactam), 1630 (pyridinium) and 1590 cm" (carboxylate); uv (HJD) : n 2 max 231 (£9800) and 307 nm (£25000); Μθ + 3.14 (c 0.5, HgO) * Example 8 Preparation of 3-^2-(3-mcthoxy-l-pyridinium)ethyl thio]6qU- [1 1 -(P)-hydroxye thyl]-7-oxo-l-azabicyclo(3.2.0) hept-2-enc-2-carboxylate A. 1-(2-mereaptoethyl)-3-methoxypyridinium methanesulfonate To precooled (5°C) 3-methoxypyridine (698 mg, 6.4 mmol) was added dropwise methanesulionic acid (0.216 mL, 3.05 mmol) and ethylene sulfide (0.19 mL, 3.2 mmol). The mixture was then heated at 60°C for 18 h, cooled to 20°C, diluted with water (10 mL) and washed with ether (3 x 10 mL). The aqueous phase was pumped under high vacuum for min and poured on a C1O reverse phase column. The 1 o title compound was eluted with water. The appropriate fractions were combined and evaporated under high vacuum to give the desired thiol (61.6 mg, yield 76.3%); ir (CII2C1,,) Vm.iy :2550 (w, SH) and 1620, 1600, 1505 cm-1 (m, aromatic); ^Hinr (DMSO (/&) δ: 8.90-7.90 (411, m, aromatic C-H), 4.72 (2H, t, 3=6.6 Hz, CH2N+), 4.01 (3H, s, OCH3), 3.5-3.0 (m, hidden CHjS), 2.66 (1H, dd, J = 9.5 Hz, £=7.5 Hz, SH) and 2.31 ppm (311, s, CH^SO^-) Β. para-Hi trobenzyl 3[2-(3-mcthoxy-1-pyridi nium chloride) ethyl thio) -6ος- [l * -(^)-hydroxyethyl j -7-oxo-l -azabicyclo (2»0)-h«.,pt-2-erH 2-car boxy I ate A cold (0°C) solution oi p-nitrobenzyl 6Α-Γΐ'-(&)hydroxyethylj-3,7-dioxo-l-azabicyclo-(3.2.0)-heptane-2carboxylate (1.04 g, 3 mmol) in acetonitrile (12 mL) was treated dropwise with diisopropylethylamine (0.63 mL, 3.6 mmol) and diphenylchlorophosphate (0.75 mL, 3.6 mmol) and stirred at 0°C for 30 min. The resulting enoi phosphate was treated with l-(2-mercaptoethyl)-3-methoxypyridinium methanesulfonate (1.14 g, 4.30 mmol) in CH3CN(7 mL), diisopropylethylamine (0.63 mL, 4.30 mmol), stirred for 30 min. and cooled at -10°C for 30 min. The solid that precipitated out of the mixture was filtered, washed with cold acetonitrile (2 mL) and dried to give the title compound (1.32 g, yield 82%); ir (nujol) V max 3320 (m, OH), 1780, 1765 (s, A-lactam C=O), 1700, 1695 (m, ester C^O) and 1520 cm"1 (s, N02) ; 1ihnr (OMSO : 9.01 (Hi, bs, H-3 aromatic), 8.75 ( IH , bd , J«5.4 Hz, H-6 aromatic), 8.35-7.95 (4H, rn, H-aromatic), 7.70 (2H, d, J=7.7 Hz, H-aromatic), 5.37 ( 2H , center of ABq, 3 = 13 Hz, CH2?NB), 5.17 <1H, d; 7=-4.9 Hz, OH), 4.87 (2H, t, J=6.3 Hz, Cll2-N®), 4.35-3.75 ( 2H , m, H-5 and H-l'), 4.00 (3H, s, 0Cil3) 3.56 (part of a t, J=6.3 Hz, CHjS), 3.5-3.20 (Jlf, m, H-6, H-3) and 1.16 ppm (3H, d, J/6.1 Hz, CH^CtlO). ί οχ C. 3- [2-(3-methoxy-l-pyridlnium)ethylthio] -β<-1_1 '-(£)hydroxyethyl]-7-oxo-l-azabicyclo(3.2.Q)-hept-2-ene2-carboxylate A solution of para-nitrobenzyl 3[2-(3-methoxy-lpyridinium chloride)ethylthio]-6cv- [l1-(R)-hydroxyethyl] -7-oxo-l-azabicyclo(3.2.0)-hept-2-ene-2-carboxylate (600 mg, 1.12 mmol) in THF (25 mL), ether (25 mL) and pH 7.4 phosphate buffer (0.1M, 25mL) was hydrogenated in a Parr shaker over 10% Pd/C (1.1 g) for 1 h at 40 psi. The mixture was diluted with ether and the aqueous phase was filtered through a ^52 hardened filter paper.

The aqueous layer was washed with ether (2 x 20 mL), pumped under vacuum and poured on a silica gel reverse phase column. The title compound was eluted with water containing 2 and 5% acetonitrile. The appropriate fractions were combined and lyophilized to give a yellow solid that was repurified by hplc togive the penem carboxylate (150 mg, 38%); ir (nujol) 1750 (s, « Π1&Χ « p-lactam C=0) and 1580 cm (s, carboxylate); 1Fmr (DgO) <5: 8.55-8.30 (2H, m, H-2, H-6 aromatic); 8.17-7.75 (2H, m, H-3, H-4 aromatic), 4.77 (2H, t, J = 5.9 Hz, CH2N®), 4.10 (IH, part of 5 lines, >6.3 Hz, H-l·), 3.97 (3H, s, OCHg) , 3.85, 3.82 (2 lines, part of dt, >2.6 Hz, part of H-5), 3.42 (2H, t, >5.9 Hz, CHg-S), 3.25 ι ο: (1Η, dd, 3=6.1 Hz, J = 2.6 Hz, H-6), 2.99-2.60 (2H, 6 lines, part of H-4) and 1.20 ppm (3H, d, J = 6.4 Hz, Cf!^) ; uv (H,,0, c 0.05)2¼ : 290 (£10517), 223 (£6643); T,, c. max /j (0.1 M pH 7.4 phosphate buffer, 37°C) 20 h. 03 Example 9 Preparation of ( 5K.; 6£)-3-[j?-(3-me thy J thiopy r idinio) ethyl thio]-6-[l-(nJ-hydroxyethyl]-7-oxo-l-axgbicycJo [3.2.0]hept-2~ene-2-carboxylate A, 3-Methylthio-l-(2-mereaptoethy1)pyridinium chloride To a solution of 3-methylthiopyridine1 (2.00 g, 0.016 mol) in 10 mL of ether was added 15 mL. of 1 N HCl and the mixture was well shaken. The aqueous phase was separated, washed with 10 mL of ether and then evaporated.

The residual hydrochloride was then dried in vacuo (PgO^) to give a white solid. To this solid hydrochloride was added 3-methylthiopyridine (1.88 g, 0.015· mol) and ethylene sulfide (0.89 mL, 0.015 mol) and the resulting mixture was heated (oil bath) at 55-65°C under for 15 h.

This gave a slightly turbid oil which was taken up in 125 mL of H^O and washed with CH^Clg. The aqueous solution was concentrated to about 25 mL and then a few drops of acetonitrile were added to make the mixture homogeneous.

The resulting aqueous solution was applied to a C ^Prepared by the method of J.A. Zoltewiez and C. Nisi, J. Org. Chem. 34, 765 (1969).

B. p-Nitrobenzyl(5R ,6S)-3- [2-(3-methylthiopyridinio)ethyl thio] -6- [l-(R)-hydroxyethyl·]-7-oxo-l-azabicyclo l3.2.θ] lO hept-2-ene-2-carboxylate chloride A solution of p-nitrobenzyl (5R,6S)-6- [l-(R)-hydroxyethyl] -3,7-dioxo-l-azabicyclof3.2.0]heptane-2-carboxylate (0.522 g, 1.50 mmol) in 7 mL of dry acetonitrile was cooled at 0°C and then diisopropylethylamine (0.287 mL, 1.65 mmol) was added dropwise. To the resulting yellowbrown solution was added dropwise diphenyl chlorophosphate (0.342 mL, 1.65 mmol) and the reaction mixture was kept at 0°C for 30 min. Diisopropylethylamine (0.313 mL, 1.80 mmol) was then added, followed by a solution of 3-inethyl t.hio-1(2-mcrcaptoethyl)pyridinium chloride (0.398 g, 1.00 mmol) in 0.70 mL of dry DMF. About a minute after the addition was complete a precipitate separated from the reaction mixture and further cooling at -10’C for 10 min gave a solid orange-coloured mass. This solid was subsequently triturated with acetonitrile and the residue was collected by filtration. The residue a og was washed with acetonitrile, then nee tone and finally dried in vacuo to give the product (¢).455 g., 55¾) ns a cream-coloured solid. The combined filtrate was evaporated to £ivc a yellow oil which was taken up in a minimum volume of acetonitrile and cooled at c). ir (KBr) V : 3545 (br, -OH), 1770 (^-lactam CO), ιΠΗλ . iu 1680 (-C02pnb) cm ; Hnmr (d^-DMSO) δ: 0.98-7.96 (m„ 4K, pyridinium aromatic), 8.20-7.65 (ABq, J=7.0 liz, 4Ho PNB aromatic)9 5.55=4.80 (ia? 4H), 4.5=5.7 (mP 2H), 5e6-5«25 (a, 6H), 2.66 (s, 5H, S-He), 1,16 (d, J=6.0 Hz, 5Π, CHMe).

C. (^65)-3::C2-(3-roe-thylthiQpyridino) ethyl thia] -6£1 -(Hj-hydre.czethyll -7-oxo-1 -azabicyclol 3._2.0]hopt-2ene~2-carboxylate To a mixture of p-nitrobenzyl (5R,6S)-3-[2-(3methylthiopyridinio)ethyl thio]-6-[ I- (R)-hydroxyethyl] -7-oxo-l-azabicyclo[3.2.0]-hept-2-enc-2-carboxylatc chloride (0.551 g, 1.0 mmol) and 107, palladium on-charcoal (0.55 g) in 25 ml of phosphate buffer (0.05 M, pH 7.4) was added 5mb of THF and 25 mb of ether. This mixture was hydrogenated (Parr) at 40 psi for 1 h„ The reaction mixture was then filtered through Celite and the filter cake was washed with H^O and ether- The aqueous phase was separated and washed with additional ether (3 x)® After removing rcsudu.il organic solvents in vacuo the adjusted to 7.0 with saturated aqueous Nahco^. This solution wag immediately applied to a Ο^θ reversephase column. Elution with HgO and subsequent lyophilization of the relevant fractions afforded Oe25 of a bright yellow solid. This material was repurified by reverse-phase hplc to give the product (0.210 g, 55½) as a light yellow solid* ir (KBr)yrri2X: 3400 (br, -OH), 1755 (^-lactam CO), 1590 (-C02") cm"1; 1Hnmr (DgO) 8.60-7.76 (m, 4H, aromatic), 4*76 (t, J=5*8 Hz, 2H, N-CH2), 4.13 (d of q, J=J’ = 6.3 Hz, 1H, H-1'), 3o95 (d of t,’ J=9.0 Hze J'=2.8 Hz, 1H, H-5), 3.45-2.75 (□, 5H), 2*59 (s, 3'H, 3-ke), 1.20 (d, J=6.4 Hz, 3H, CHHe); uv (H2O)Xmax: 296 (£S5C9), & UltiA 273 (£13005), 231 (fi1576) nn; (pH 7.4, 36.8°C) 20 h. Ex.-un pie 10 p£Cj>arp.tj.on of 3- Γ?-( 1-,L2.Qrdi_me thylpy ?;id in ivun )ethy 1 thio] -6«- Q -(E) -hydro xye thy -?-o_vo-1 azabl cyclo (3 2.0)hept-2-ene-2-carbQxyl_ate coo A. 1-(2-mereaptoethy])-2,6-d ime thylpyridinium methanesulfonate A mixture of 2,6-dimethylpyridine (19.2 mL, 0.165 mol) and methanesulfonic acid (3.27 mL, 0.050 mol) was stirred for 15 min, treated with ethylene sulfide (4.17 mL, 0.070 mol) and stirred at 100° C for 42 h under a nitrogen atmosphere. After cooling to 25eC, the reaction mixture was diluted with ether (45 mL) and water (30 mL). The two layers were separated and the organic layer was extracted with water (2x5 mL).

The aqueous layers were combined, filtered through a Celite pad, washed with ether (2 x 15 mL), pumped to remove the traces of organic solvents and poured on top of a column (3.0 x 12 cm) of p-bondapak C-18.

Elution with 3% acetonitrile 97% water mixture gave after lyophylization of the appropriate fractions 2.5 g of the impure title compound as syrup. It was repurified by hplc (ji-bondapak C-18) to give 0.90 g (7%) of the title compound. ir (film) 0 : 2520 (SH) max . 1640 and 1625 (pyridinium), 1585, 1490, 1200 cm" (sulfonate) 1Hnm (DMSO-c/θ + DgO) 6: 2.36 (3H, s, CH3SO3): 4.62 (2H, in, CII2N + ) . 7.74 (2H, m, Hm of pyridinium), 8.24 (1H, m, Hp of pyridinium), uv (H^O) max: 272 (£4080) mp 110 B. Parani trobenzyl 3- [2-(1-( 2,6-dimethylpyridinium)) ethylthio-6K- [~1-(H)-hydroxye thy i] -7-oxo- i -nzabicycio (3,2.0)hept-2-ene-carboxylatc diphenylphosphate To a cold (0°C) solution of p-nitrobenzyl 6^-[l(]j)-hydroxye thyl]-3,7-dioxo-l-azabieyclo( 3.2.0) hep tane2-carboxylate(0.658 g, 1.89 rnmol) in acetonitrile (6mL) kept under a nitrogen atmosphere was added diisopropylethylamine (0.394 mL, 2.25 mmol) and diphenyl 1θ chlorophosphate (0.468 mL, 2.26 mmol). The reaction mixture was stirred 30 min and treated with a solution of l-(2-mercaptoethyl)-2,6-dimethylpyridinium methanesulfonate (0.720 g, 2.73 mmol) in acetonitrile (3 mL) followed by diisopropylethylamine (0.394 mL, 2.26 mmol).

The reaction mixture was stirred at 0°C for 2 h, diluted with c^ld (0°C) water (27 mL) and poured on top of a column (2.5 x 9.0 cm) of μ-bondapak C-18. Elution with acetonitrile-water mixtures and lyophylization of the appropriate fractions gave 0.92 g (65%) of the title compound, ir (KBr) \) : 3700-3000 (OH), 1765 max (C=0 of /3-lactam) , 1690 (C=o)of PNB ester), 1620 (pyridinium), 1590 (phenyl), 1517 (N02), 1330 (N02), 880 cm-1 (N02), ^mr (DMSO.c/θ) 5: 1.15 (3H, d, J=6.2 Hz a. ϊ i ClLjCHOH), 2.7-3.7 (11 H, CHjS, 2-CH3 on pyridinium, H-4, H-6), 3.7-4.4 (211, CHgCHOH, H-5), 4.7 (2H, m, CH2N+), 5.14 (1H, d, £=4.5 Hz, OH), 5.37 (center of ABq, £=13.2 Hz, CHg of PNB), 6.7-7.5 (10H, m,phenyl), 7.5-8.7 (7H, pyridinium, H's of PNB), uv (HgO) >max: 274 319 (£9^45) mp C. 3- [2-(1-(2,6-dimcthylpyridinium)c thylthio]-6Λ- [l(fl)-hydroxyethyl]-7-oxo-l-azabicyclo(3.2.0)hcpt-2-ene2-carboxylate To a solution of p-nitrobenzyl 3-E2-(1-(2,6-dimethylpyridinium))-ethylthio] -6^- [l-(R)-hydroxyethyl7-oxo-l-azabicyclo(3.2.0)hept-2-ene-2-carboxylate diphenylphosphate (0.80 g, 1.07 mmol) in wet tetrahydrofuran (42 mL) was added ether (42 mL), 1θ potassium phosphate monobasic-sodium hydroxide buffer (0.15M, pH 7.22, 21 mL) and 10% palladium on charcoal (0.80 g). The resulting mixture was hydrogenated for 1 h under 40 psi at 23°C and filtered through a Celite pad. The two layers were separated.and the organic layer was extracted with the buffer (3x3 mL). The aqueous phase were combined, washed with ether (50 mL), pumped to remove traces of organic solvent and poured on top of a column (3.0 x 10.2 cm) of μ-bondapak C-18. Elution of the column with 5% acetonitrile - 95% water mixture and lyophylization of the appropriate fractions gave the title compound 0.246 g (63%) as a yellowish powder, ir (KBr) \) max: 3700-2800 (OH), 1750 (C=0 of A 13 the /3-lactam), 1620 (pyridinium), 1585 cm (carboxylate), ^Hmr (D^O) S: 1.23 (3H, d, J = 6.4 Hz, CHgCHOH), 2.5-3.5 (11H, H-4, H-6, CH2S, 2CH3 on pyridinium), 3.8-4.4 (2H, CH3CHOH, H-5), 4.5-4.9 (CH2N+ HOD), 7.64 and 7.74 (2H, A part of A2B system,Hm of pyridinium), 8.07, 8.16, 8.18 and 8.27 (lH, B part of A,B system, Hp of pyridinium), uv (H?0) X : 277 (€9733), 300 (68271) mjA, 0·]β + 50.7 (C 0.48, H20), Anal. calcd. for cl8H22N204S’1 *5 H20: C 55’51» H 6.47, N 7.19; found: C 55.14, H 6.23, N 6.46. ί .1 '3 Example 11 Preparation of (53/ 6g)-3-(2-(2-pctbyjttlo-3methylimidazolio)ethyl-thiol-6- tl- (§) -hydroxyethyl]-7-oxo-l-azabicyclp[3.2.0]hept-2-ene- A. 2-Methylthio-3-methvl-l-(2-mercaotoethy1)imtdazoliun trifluoromethanesulfonate /CX + cr3S033 + Δ I Me Trifluoromethanesulfonic acid (1,38 mL, 0.015 mol) was added dropwise to 2-methylthio-l-methylimidazole^ (4.0 g , 0.03 mol) at 0°C under N2· Ethylene sulfide (0.9 mL, 0.015 mol) was then added and the mixture was heated at 55*C under N2 for 24 h. The reaction mixture was triturated with ether (3x) and the residue was taken up in acetone, filtered and evaporated. This gave the product (4.2 g, 82%) as a semicrystalline solid which was used as such without further purification. ir(film) v ; 2550 (w, sh) cm \ ϊ-Hnmr max (dg-acetone) ¢: 7.97 (s, 2H) , 4.66 (t, J=7 Hz , 2H, methylene) , 4.17 (s, 3H, N-Me), 3.20 (d of t, J=7 Hz, J'«9 Hz, 2H, methylene), 2.72 (s, 3H, S-Me), 2.20 (t, J=9 Hz, 1H, -SH). 1. Prepared as per A. Wohl and W. Marckwald, Chem. 3er. 22, 1353 (IS 89). Β· p-Nitrobenzyl (5g, 6S)-3-( 2-(2-methvlthio-3-methy1imidazolio) ethyl thio]*6= fl- (R) -hydroxyethylj-7-oxol-azabicyclo [3. 2 .0]hept-2-ene-5-carboxylate diphenylphosphate -> To a solution of p-nitrobenzy1 (5R, 6S)-6-[ 1-(R) hydroxy ethyl] - 3,7-dioxo-l-azabicyclo[3.2.0]heptane-2-carboxylate (1.40 g, 4.0 mmol) in 50 mL of dry acetonitrile# at 0eC under was added dropwise diisopropylethy lamine ¢0.76 mL, 4.4 mmol) followed by diphenyl chlorophosphate (0-91 mL, 4.1 nmol).

After stirring the reaction mixture at room temperature for 1 h, diisopropylethylamine (0.76 mL, 4.4 mmol) was added and then a solution of 2-methylthio-3-methyl-l-(2-nercaptoethyl)imidazolium trifluoromethanesulfonate (2.0 g, 5.9 mmol) in 5 mL of acetonitrile was added dropwise. The reaction mixture was kept at room temperature for 1.5 h and was then concentrated in vacuo to give a gum. This gum was taken up in E^Q and applied to a C-jg reverse-phase column. Elution with H^O, then 20¾ acetonitrile-H^O and finally 30¾ acetonitrile-H^O, followed by lyophilization of the appropriate fractions afforded the product ¢0.90 g, 30%) as a light yellow solid. ir(K3r) v : 3380 (br, OH), 1770 (β-lactam CO) -1 ι max cm ; *Hnmr (dg-acetone) 6: 8.35 (br,s, 1H), 8.24, 7.78 (A3 q, J = 8.8 Hz, 4H, aromatic), 7.89 (br s, 1H), 7.25-6.91 (m, 10H, diphenylphosphate), 5.50, 5.25 (A3 q, u=12 Hz, 2H, benzylic), 4.75-4.27 (m, 3H), 4.03 (s, 3H, N-Me), 4.15-2.75 (m, 8H),. 2.59 (s, 3H, S-Me), 1.22 (d, J=6.2 Hz, 3H, -CHMe).

C. (55, 65)-3-( Z-ZZ-Methylthio-S-methylimidazoliojethylthio)-6-[1-(R)-hydroxyethyl]-7-oxo-l-agabicyclo[3.2.0]heDt2-ene-2-carboxylate To a solution of p-nitrobenzy1 (5R, 6S)-3-[ 2(2-methyl thio-3-methylimidazolio)ethyl thio]-6- (1-(R)hydroxyethyl]-7-oxo-l-azabicyclo[3.2.0]hept-2-ene-2carboxylate diphenylphosphate (1.20 g, 1.56 mmol) in a mixture of 70 mL of TH?, 70 ml of ether and 31 mL of 10 phosphate buffer (0.05 M, pH 7.4) was added 1.2 g of 10% palladium-on-charcoal. This mixture was hydrogenated (Parr) at 35 psi for 55 min. The reaction mixture was then filtered through Celite and the filter cake was washed with H^O ether. The aqueous phase was separated, cooled at 0eC and the pH was adjusted to 7.0 with saturated aqueous NaHCOj.

After removing residual organic solvents in vacuo the aqueous solution was applied to a θ reverse-phase column. Elution with H^O and then 3% acetoni-rrile-H^O end subsequent lyophilization of the .relevant fractions gave 0.25 g of a 20 solid. This material was repurified by reverse-phase hplc to give the product (0.114 g, 19%) as an off-white solid. ir(K3r) v : 3420 (OK), 1750 (8-laetamCO), 1590 (-CO-®) cm-1; max 2 'Kn-ur (D2O) S: 7.58 is, 2H), 4.52 (t, J=6 Ha, 2H), 4.283.82 (m, 2H), 3.90 (s, 3H, N-Me) . 3.40-2.87 (m, 5H) , 2.40 (s, 3H, S-.Me), 1.20 (d, 0 = 5.4 Hz, 3H, -CHMe) ; uv (H-0) λ_ : ΓΓι ax 297 (ε 7572), 262 (ε 6259), 222 (ε 7955) nm. 117 Example 12 Preparation of <5g, 6j5)-3-[ 2-(3-aminopyridinio)ethy1 thio]- 6-[1~ -hydroxyethyl]7-oxo-l-azabicyclo[3.2.0)hept-2-ene-2-carboxylate A. ?-Απάπο-1-(2-mercaptoethy1)pyridinium chloride 3-Aminopyridine (1.50 g, 0.016 mol) was taken up in 15 mL of λ N methanolic HCl and the resulting solution was evaporated to give the hydrochloride as an oil· To this oil was added 3-aminopyridine (1.32 g, 0.015 mol) and ethylene sulfide (0.89 ml, 0.015 mol) and the resulting mixture was heated (oil bath) at 60-65°C under for 2 h. . Another equivalent of ethylene sulfide ($.89 ml, 0.015 mol) was added and heating was continued at 55-65°C for 65 h.

The reaction mixture was washed with CH2C1O and then taken up in H0O (25 ml). The aqueous solution was applied to a reverse-phase column which was eluted with H2<0.

Evaporation of the relevant fractions gave the product (1.26 g 44%) as a colorless, viscous oil. ir(film) v : 3180 (NH-) max 4 cm; hhmr (d^-DESO) ό: 8.19-7.59 (m, 4H, aromatic), 4.59 (t, M2 Hz, 2H, N-CH-) 3 5 (hr s, 2H, -NH-) , 3.20-2.77 *— <ί & (m, 3H). . p-Nitrobenzyl (53» 65)-3-( 3-(3-amiπο?νridiniο)ethy1 thio) -6-( 1~(E) - hydroxvethyI)-7-oxo-azablcyclo( 3.2. 0) To a solution of p-nit rob ensyl (53, 6s_)-6-( 1" (R) by drdxyethyl) - 3 » 7- di oxo-1-azabi cyclo ( 3.2.0 )hept ane-2carboxylate (Ο.696 g, 2.0 nmol) in 10 nL of dry acetonitrile, at 0°C under N^» Vas &dded dropwise diisopropylethylanine (Ο.382 aL, 2.2 nmol) followed by diphenyl chlorophosphate (0.1*57 mL, 2.2 aaol). After stirring at 0°C for 30 min- a solution of 3-anino-1-(2-aercaptoethy1)pyridiniua chloride (O.U75 g, 2.5 MOl) in 1 ml of dry DMF was added, followed by additional diisopropylethylamine (0-1*35 mL, 2-5 mmol).

The reaction mixture was kept at 0°C for 1.5 h and was then concentrated in vacuo. The resulting gum was taken up in aeetonitrile-K„O (1:1) and applied to a C,0 rev·rse-phasc 2’ ------------------- _ _l8 column- Sluti and subsequent lyophilization of the relevant fractions afforded the product (0.730 g, 50?) as a beige-colored solid. ir(K3r) v : 3330 (br, OH), 3180 (br, HH,), 1770 (S-lactaa CO), BIX . . — 1690 (-C0 FB3)ca ; 1Hnnr (dg-DMSO) δ: 8.29-7-63 (=, 8 iroaitic), 7.2-6.7 (b, 10H, diphenylphosphate), 5 · *· 7 , 5·1δ (A3 q,· J= lh Hz, acetonitrile-H^O 2H, benzylic), 1». 73-U.U5 (η, 3H) , 1(.2-3.8 (a (a, 8h), 1.15 (d, J"6.2 Hi, 3H, CHMe).

IH), 3.6-2.6 119 C . (55, 65)-3-( _2-( 3-Aminopyri di nio ) e tnyl _ ( (R) -hydroxyethyl )-7- oxo-l-a2abicyc lo ( 3.2., 0 )hept-2ene-2-carboxylate To a mixture of p-nitrobenzyl (5£> 6s)-3( 2-(3aminopyridiaio)ethyl thio)-6- (l- (R) "hydroxyethyl)-7-oxo-l~ azabicyclo(3.2.0)he?t-2-ene-2-carboxyl ate diphenylphosphate (0-730 g, 1.0 mmol) and 10? palladiua-oa-charcoal (0.7 g) in 25 air of a phosphate buffer (0.05 M> p3 7-^) vas added 8 mL of TE? and 20 mL of ether. This mixture vas then hydrogenated (?arr) at ho psi for 1 h. The resulting mixture vas filtered through & pad of Celite and the filter cake vas vashed vith H^O and ether. The aqueous phase was separated, washed vith ether (2 x) and then residual volatiles were removed in vacuo. The aoueous solution vas immediately applied to a C^q reverse-phase column vhich vas eluted vith SgO· Lyophilization of the relevant fractions afforded 0.1*5 g of an off-vhite solid. This material vas repurified by reverse-phase hplc to give the desired product (0.123 g> ^) as an ivory-colored solid. ir(X3r) v : 33^0 (br), 1750 (br, δ-lactam CO), 1580 (br, o sax -COg ) cb'1; 'Hnar (DgO) δ: 8.07-7.59 (π, ** Η, aronatic), b.6l (t, J; 5.8 Hz, 2Ξ, N-CB,), b. lU (d of q, J=J’^6.3 Hz, In, H-l'), 3.97 ( d of t, Js?.2 Hs, J'~2.6 Hz, 13, H-5), 3.38 (t, J»5.8 Hz, 23, S-CHj), 3.2^ (d of d, J=6.0 Hz, J’ = 2.6 Hz, 1H, H-6), 3.17-2.57(0, 2H, H-M, 1-21 (d, J=6.3 Hz, 3H, CHMe); uv{320) lBax: 299 (ε 791-9), 256 (ε 8822) no; t,/2 (pH 7. b, 36.8’c) 18.5 h. no Example 13 ί 5R, 6S) 3- [Ϊ- (S)-methyl-2- (l-pyridiniuaa) ethylthio] -6- II- (*)5 hydroxyethyl]-7-oxo-l-azahicyelot3.2.0)hept-2-ene-2-carboxyla and (5R,6S) 3-[1-(R)-methyl-S-(1-pyridinium)ethylthio)-6-[1-(R)hydroxyethy1]-7-oxo-l-aaabicyclo[3.2.0)hept-2-ene-2-C£rboxyie A. d£-l-(2-nercapto-2-methylethyl)pyridinium methanesulfonate dJL-l- (2-nercapto-l-methyIethyl)pyridinium methanesulfonate Ms OH.

Methanesulfonic acid (1.95.mL, 0.030 mol) was added slowly Lo cold pyridine (7.83 ml, 0.097 mol) and the resulting mixture was stirred at 40°C for IS min, treated with d£-propylenesulfide (2.59 mL, 0.033 mol) and stirred at 606C under a nitrogen atmosphere tor 90 h. Pyridine was removed under vacuum; the residue was mixed with water and purified by chromatography (hplc, Prep. Bondapak C-18). Tne appropriate fractions were combined and lyophilized giving d£-l-(2-mercapto-2-methylethyl)pyridiniua methanesulfonate 1,14 g (15¾) as a colorless syrup? ir (film) v : 2S20 (SH), 1640 (oyridinium), 1180 (S, GLSo/), 1040 max 3 3 (CH,S0)ca\ ’Hmr (DMSO <) δ: 1.35 (d, >6.8 Hz, 33, CH,CH5) , 2.30 's, 3H, O^SO-), 2.90 (d, J-8.5 Hz, IH, SH) , 3.2-3.7 (n, CKSK) , 4.52 (dd, J "12.9 Hz, >8.4 Hz, CHCH.N+) , 4.37 (dd, J »12.9 Hz, J-6.0 Hz, CHCrJ N+) —gem — —2 —gen — —2 8-0-8.4 (m, 2H, Hm of pyridinium), 8.5-8.8 (m, IH, Hp of pyridinium), 9.04 (dd, J»1.4 Hz, J-6.7 Hz, 2H, Ho of pyridinium), uv (HO) λ : 208 (c5267), 259 (£3338), Anal, calcd. for C9H15^°3S2* 2H2°« c 37.88, H 6..71, N 4.91, S 22.47; found: C 37.49, H 6.85, N 4.86, S 22-09 and dt-1- (2-mercaoto-lfl: methylethyl) pyridinium methanesulfonate 0.32 g. <11%) as a colourless syrup; ir (film) 2500 (SH) , 1628 (pyridinium)., 1180 (sulfonate, 1035 (sulfonate) cm \ 3Kmr (DMSO £^) 5: 1.69 (d, £=*6.8 Hz, 3H, •^3CHN+b 2,31 fs' 3H' 3-0-3-3 (η, 2H, CH^S) , 4.2-5.2 (m, Γ) CBN )' 8.0-8.4 (m, 2H, Hm of pyridinium), 8.5-3.3 (η, 1H, Kp of pyridinium), 9.0-9.2 (m, 2H, Ho of pyridinium), uv (H_0) λ : 209 2 max (C4987), 258 (C3838). Anal, calcd. for CH NO S ·1.5Η_0: C 39.11, 32 2 H 6.55, N 5.07; found: C 39.13, H 5.92, N 5.20.

B. (5g, β£) paranitrobenzyl 3-11-(R,$)methyl-2- (l-pyridiniura) ethylthio)-6I 0 ii- (R)-hydroxyethyl)-7-oxo-1-agabicyclo(3.2.0)hept-2-ene-2=carboxylate diphenylphosphate 4) NEtfiPr)^ To a cold (0eC) solution cf (SR, 6£) paranitrobenzyl 6-(1-(2)hydroxyethyl]-3,7-dioxO-1-azabicyclo (3.2.0) he?tane-2-carboxyIate I > (0.523 g, 1.5 mmol) in acetonitrile (6 mL) kept under a nitrogen atmosphere was added diisopropylethyl&mine (0.314 nL, 1-8 jczdo!) followed by diphenyl chlorophosphate (0.373 mL, 1.8 mmol). The reaction mixture was stirred for 30 min and treated with a solution of d£-l-(2mercapto-2-’fnethylethyl)pyridinium methanesulfonate (0.539 g, 2.16 mmol) in acetonitrile (2 mL) and diisopropylethylamine (0.314 mL, 1.8 mmol).

The reaction mixture was stirred at 0*C for 1 h, diluted with cold (0®C) water (24 mL) and chromatographed over prep bondapak C-18 column (2.5 x 8.5 cm) with 25-50¾ acetonitrile in water as elutiDg solvents to give 1.07 g (97¾) of the title compound as a yellowish powder after lyophilization; ir (X3r) V : 3700-3100 (OH), 1770 (C=O of β-lactam), max 1695 (C"OofPN3 ester),. 1630 (pyridinium), 1S90 (phenyl), 1S18 (N02), 1348 (NOj) , 885 (NOj) cm1, 'hdiT (DHSO d ) 6: 1.14 (d, J-6.1 Hz, 3H, CHjCHO), 1.33 (d, £"6.3 Hz, 3H, CHjCHS), 4.6-5.0 (m, O^lT) , 5.14 (d, £=5.2 Hz, IH, OH), 5.37 (center of ASq, £-12.4 Hz, 2H, CH, of PN3), 6,6-7.5 (m, 10H, phenyl of phosphate), 7.69 (d, £«*8.7 Hz, 2H, Ho of ?N3) , 8.0-8.4 (m,-4H, rim of PN3,· Hm of pyridinium), 8.4-3.8 (m, IH, H? of pyridinium), 9.08 (d, J5.6 Hz, 2fi, Ho of oyridiniCT), uv (HO) λ : — ‘ 2 max 263 (£13325), 308 (£8915). Anal, calcd. for N 0 S?· HO: CS7.S2, ' · 3 b 3 b 3 10 2 H 5.10, N 5.59, S 4.27; round; C 57.76, It 4.96,N5.36, S 4.35.

C. (5g,65) 3- (1- (R and S) -methyl-2- (1-pyridinium) ethylthio) -6- fl- (R) hydroxyethyl)-7-oxo-l-az&bicyclo(3.2.0)hept-2-ene-2-carboxv late s> > ^r-t-X i-.W THF, ether, buffer I coo* 2- (1-pyridinium) ethylthio]-6- [1- (R) -hydroxyethyl)-7-oxo-l-azabicyclo(3.2.0)hept-2-ene-2-carhoxylate diphenylphosphate (0.60 g, 0.82 mmol) in wet tetrahydrofuran (33 mL) was added ether (33 mL) , potassium phosphate mono basic-sodium hydroxide buffer (17 mL, 0.15N, pH 7.22) and 10¾ palladium on charcoal (0.60 g). The resulting mixture was hydrogenated for 1 h under 40 psi at 23*C. The two layers were separated and the I :: -1 organic layer vas extracted with water (3x7 mL) . The aqueous layers were combined, filtered through a Celite pad, washed with ether (3 x 20 mL) and chromatographed on prep bondapak C-18 column (2.5 x 9.5 cm) with water as eluting solvent to give 0.18 g (63%) of mixture of diastereoisomers.

The two diastereoisomers were separated by hole (prep bondapak C-18) with water as eluting solvent: isomer with' Iowa- retention time, 0.068 g (23\) compound B, ir (X3r) V : 1770 (OO of β-lactam), 1633 (pyridinium), 1593 max (carboxylate) cm \ ^Hnur (D^O) 6: 1.20 (d, £^6.3 Hz, 3H, CH^CHO), 1.42 (d, 3=6.9 Hz, 3H, CHECKS), 2.3-3.2 (m, 3H, H-4, H-6), 3.5-3.9 (ra, 1H, SCH) , 3.9-4.2 (m, 2H, H-5, CH^CKO), 4.3-5.1 (m, G^N*}, 7.8-8.2 (ra, 2H, Km of pyridinium), 8.4-8.7 (m, 1H, Hp of pyridinium), 8.7-9.0 (ra, 2H, Ho of pyridinium uv (HO) λ : 260 (E6727), 300 (E824S),. [dl 23-.39.3* (c, H,0) , T, -12.6 h * max D — 2 % -4 ** (measured at a concentration of 10 M in phosphate buffer pH 7.4 at 36.8°C)i isomer with higher retention time, 0.081 g (28%), compound A, ir (K3r) v : 1755 (OO of β-lactam), 1630 (pyridinium) , 1590 (carboxylate) max cm \ lHcu- (DO) <5: 1.18 (d, .£=6.3 Hz, 3H, CH CHO), 1.40 (d, J«7.0 Hz, 3H, CHECKS), 2.84 (d, £=9.3 Hz, 2H, H-4), 3.26 (dd, £"2.7 Hz, £*5.9 Hz, 1H, H-6), 3.4-4.2 (m, 3H, SCH, O^CHO, H-5), 4.2-S.l (ra, C3^N+) , 7.7-8.1 (m, 2H, Hm of pyridinium), 8.3-8.65 (ra, 1H",‘· Hp of pyridinium) , 8.65-8.9 (ra, 2H, Ho of pyridinium), uv (H^O) λ : 259 (E5694), 296 (ε6936), max Id) . -*96.9® (c 0.56, H O), τ. *15.6 h (measured at a concentration £ - 2 2 . *4 of 10 H in phosphate buffer pH 7.4 at 36.8®C). ι;;5 Example 14 PREPARATION ΟΓ (5R, 6£) 3-(2-( (S>- (1-pyridiniura) ]-l- (S)-cyclohexylthio)-6[1- (3)-hydroxyethyl)-7-oxo-l-axabicyclo(3.2.0)hept-2-ene-2carboxylate and (58,65)3-(2-I(8)-(1-pyridinium))-1-(R)-cyclohexylthio]-6-(1-(R) hydroxy ethyl)-7-oxo-1-azabi.cyclo (3.2.0)hept-2-ene-2-carboxylate rjG A. c£Z"l- [2-mercapto-l-cyclohexyl)pyridinium methanesulfonate Tj Hl I i . i - . . .1 ..... . I I I - ' — - I . ! I — i — W Methanesulfonic acid (0.65 mL, 0.01 mol) was added dropwise to pyridine (2.42 mL, 0.03 mol) with cooling. The mixture was stirred 5 under a nitrogen atmosphere for.10 min, treated with c£Z-cyclohexenesulfide [1.377 g (85¾ pure), 0.0102 mol) and stirred at 72*C for 25 h.

The excess of pyridine was removed under vacuum and the traces were codistilled with water. lhe residue was mixed with water and chromatographed, through prepbondapak C-13 column (5 x 13 cm) with 0-2¾ acetonitrile 0 £n water as eluting solvent giving after lyophilization a colourless syrup 1.57 g (53%), ir (film) V : 2500 (SH) , 1625 (oyridinium), 1190 max '.SO (DHSOd) δ: 1.2-2.5 (=, 8H, cyclohexyl H) , 2.32 (s, 3H, 6 CH3SO3"), 2.82 (d, £=9.8 Ha, SH), 3.0-3.5 (m, 1H, CHSH), 4.2-4.9 (m, 1H, + CKN ), 8.0-8.3 (m, 2H, Hm of pyridinium), 8.4-8.8 (η, 1H, Hp of pyridinium) 1 8.9-9.3 (m, 2H, Ho Of pyridinium), uv (HO) λ ; 214 (£5365) , 258 (63500). max Anal, calcd. for ' H20; c <6-83* H 6.88, N 4.56; found: C 46.61, H 6.46, N 4.65. r; 7 B. (5R,6S)paranitrobenzyl 3-[2-[(Ror S)- (l-pyridinium)]-1-(R or S)-cyclohgxylthioJ-6-(1-(R) -hydroxyethyl]-7-oxo-l-azabicyclo (3.2.0)he?t-2-ene-2-carboxylate diphenylphosphate r> To a cold (0*C) solution of (5R, 6_S)paranitrobenzyl 6-(l-(R)~ hydroxyethyl]-3,7-dioxo-l-azabi'cyclo£3.2.0)heptane-2-carboxylate '(1.37 S, 3.93 mmol) in acetonitrile (15 mL) kept under a nitrogen atmosphere was added diisopropylethylamine (0.822 mL, 4.7 mmol) and diphenyl chlorophosphate (0.979 mL, 4.7 mmol). The resulting solution was stirred for )0 30 min and treated with a solution of ^f-1-(2-raercapto-l-cyclohexyl)pyridiniu7?? methanesulfonate (1.64 g, 5.66 mmol) in acetonitrile (4.7 mL) followed by diisopropylethylamine (0.822 mL, 4.7 mol). The reaction mixture was stirred at O’C for 1 h, diluted with cold (0*C) water (75 mL) and chrcmztographed on prepbondapak C-18 with 25-50¾ acetonitrile in water as eluting '3 solvent giving after lyophilization of the appropriate fractions 1.9 g (53¾) of the title compound, ir (K3r) v : 3700-3000 (OH), 1770 (C=0 of max. β-lactam), 1700 (CO of ?NB ester), 1628 (pyridinium), 1590 (phenyl), 1515 (NO^), 1345 (NO,), 880 (NO,) cm1, ’Hmr (D^O) 6: 1.13 (d, >6.1 Hz, 3H, CKjCHO), 1.2-2.S (m, 8H, cyclohexyl K), 2.7-3.5 (m, 4H, H-4, H-6, CHS), 3.5-4.4 (ra, 2H, CHjCHO, H-5), 4.4-5-0 (m, IH, CKNj, 5.30 (center of A3q, >12.8 Hz, CH2 of PN3). 6.7-7.4 (m, 10H, phenyl) , 7.65 (d, >8.6 Kz, 2H, ι β Ho of PiQ) , 7.9-8.4 (m, 4H, Hm of ?N3, Hm of pyridinium), 8.4-8.8 (m, 1H, Hp of pyridinium), 9.0-9-4 (η, 2H, Ho cf pyridinium), uv (3^0) λ : 263 (E9038), 309 (E6394). Anal. calcd for C, H NO Ξ?·HO: max — — 39 40 3 10 2 C 59.16, H 5.35, N 5.31; found:' C 53.95, 8 5.15, N 5.57.

C. (sjL6£> 3- [2- [ (R or S)-(1-pyridinium) )-1-(g. or Sj-cyclohexylthio]-6[1- (R)-hydroxyethyl)-7-oxo-l-azabicyclo(3.2.0)hept- 2-ene-2-carboxylate To a solution of (5R,6g) paranitrobenzyl 3-[2-((g or S) (1-pyridinium))-1-(3 or S}-cyclohexylthio]-6- (1-(3)-hydroxyethyl)-7oxo-l-azabicyclo(3.2.0)hept-2-ene-2-carboxylate diphenylphosphate (1.85 g, 2.34 mmol) in wet tetrahydrofuran (96 mL) was added ether (96 mL), potassium phosphate monobasic-sodium hydroxide buffer (0.15M, pH 7.22, 50 mL) and 10% palladium on charcoal (1.9 g). The resulting mixture was hydrogenated at 23" under 40 psi for 1.25 h. The organic layer was separated and washed with water (3 x 20 mL). The aqueous solutions were filtered through a Celite pad, washed with ether (2 x 60 mL) , pumped to remove the traces of organic solvents and chromatographed on prepbondapak C-18 column (4.5 x 9 cm) with 0-5% acetonitrile in water as eluting solvent giving after lyophilization 0.705 g (76¾) of a mixture of diastereoisomers. The separation of the diastereoisomers was done by hplc (prepbondapak C-18) with 4% acetonitrile in water as eluting solvent; diastcreoisomer with lower retention txme, compound λ , (0.29 31%), ir (K3r) v : 1750 (C=>0 of β-lactam), 1620 (sh, pyridinium), 1685 max (carboxylate) cm \ 'Hmr (D^O) 6: 1.21 (d, £’6.3 Hz, 3H, CT^CHO) , 1.4-2-5 (m, 8H, cyclohexyl H), 2.5-3,05 (m, 2H, H-4), 3.05-3-25 (m, lH, H-6), 3.33.7 (m, 1H, CHS), 3.9-4.3 (m, 2h, H-5, CHjOjO), 4.3-4.8 (m, CHN ), 7.88.2 (m, 2H, Hm of pyridinium), 8.3-8.7 (m, 1H, Hp of pyridinium), 8.8-9.1 (m, 2H, Ho of pyridinium), uv (Ho0) λ : 260 (ε7123), 300 (C8685), i max fa] +6.2" (£ 0.63, H^O) , «16.6 h (measured at a concentration of M m phosphate buffer pH 7.4 at 36,8*C); Anal, calcd, for c2oH24N2°4S •2H2O: C 56.59, H 6.65, H 6.60, S 7.55; found: C 56.83, H 6.47, N 6.59, S 7.43; diastereoisoaer with higher retention tine, compound 3", (0-35 38%) ir (K3r) 1750 (C=0 of 6-lactam) , 1622 (sh,' pyridinium), 1588 (carboxylate) cm 1r ,Hmr (1^0) 6; 1.19 (d, £’6.4 Hz, 3H, G^CHO), 1.3- 2.5 (m, 8H, cyclohexyl Η), 2.5-3.1 (m, 2H, H-4),' 3.1-3.3 (m, 1H, K-6), 3.3- 3.8 (m, 23,H-5, CHS), 4.1 (center of m, Uh CH CjO),. 4.25=4.7 (ra, 12, GiN ), 7.8-8.1 (m, 2K, Hm of pyridinium), 8.3-8.7 (m, 1H, H? of pyridinium), S. 75-9.0 (m, 2H, KO of pyridinium), uv (HO) 1 : 259 (65992), 296 (C7646), max 23* -a faJ^ *65.3* (£ 0.43, H^O), Ti "20.2 h (measured at a concentration of 10 in phosphate buffer pH 7.4 at 36.8*C).

Example 15 A,. (SRI Allyl 3-( (2-pyridinioethyl) thio}- (6Ξ)- ((IR) -hydroxyethyl ] -7-oxo1-azabicyclo(3.2.0]hept-2-ene·2-carboxylate diphenylphosphate To a solution of (SR) allyl 3,7-dioxo~(6S)-[(lR)-hydroxyethyl]1-azabicyclo[3.2.0)heptane-(2£)-carboxylate (473 mg, 1.87 mmol) in CH^CN (6 mL) was added at ca. -10®C under a nitrogen atmosphere diisopropylethv1amine (0.42 mL, 2.4 mmol) followed by diphenyl chlorophosphate (0.50 mL, 2.4 mmol). The mixture was stirred at 0®C for 30 min, and then cooled to -15®C. To this was added an oily suspension of N-(2-mercaptoethyl)pyridinium chloride (527 mg, 3.00 mmol) in CH^CN (1 mL) containing 5 drops of DMF, followed by diisopropylethylamine (0.42 mL, 2.4 mmol). The mixture was stirred at -15® for 30 min and then diluted with H^O (20 mL)· This mixture was directly purified on a reverse phase silica gel (C^g PrepPAX, 12. g, Waters Associates) eluting with H^O (200 nL), 10% CH^CN/H^O (100 mL), 20% CS^CN/F^O (100 mL) , 30% O^CN/I^O (100 mL) and then 40% CH^CN/H^O (100 mL). Appropriate fractions were collected, the organic solvent removed by a vacuum pump and lyophilized to obtain 786 mg (1.26 mmol, y. 67.3%) of the title compound as brownish powder: 1Hnr (DMSO-<. CFT-20) 5: 1.16 (3H, d, >6 Hz, 1*-CH_), 2.6-3.7 (n) , 3.75-4.3 (2h, m, 5-H and l'-H), 4.65 (2H, m, -CO2CH2~>» 4’87 (2Hf -=6 Hz' 5-6.2 (3H, m, olefinic protons), 6.6-7.4 (m, aromatic protons), 8.15 (2H, t, 3=7 Hz, aromatic protons meta to the nitrogen), 3.63 (IH, "t, j-7 Hz, aromatic proton para to the nitrogen) and 9.07 ppm (2H, *dH, £-7 Hz, aromatic protons ortho to the nitrogen); ir (film) V: 3400 (OH), 1770 (8-lactam), 1690 (ester), 1625 (pyridinio)· B. (5R) 3-((2-pyridinioethyl)thio)-(6S)-t(IR)-hydroxyethyl)-7-oxo-lazabicyclo[3.2.0)hept-2-ene-2-carboxylate Pd(P$3)4 ΡΦ. ®ΟΡΟ(Οφ)2 CO^K To a solution of (5R) allyl 3-[ (2-pyridinioethyl) thio] - (65) ί(IR)hydroxyethyl)-7-oxo-1-azabicyclo [3.2.0)hept-2-ene-2-carboxylate diphenylphosphate (156 mg, 0.25 mmol) in CH^CN (2 mL) was successively added at ca. 22°C a solution of potassium 2-ethylhexanoate in EtOAc (0.5 M, 0.6 mL; 0.3 mmol), trlphenylphosphine (15 mg, 0.057 mmol) and tetrakistriphenylphosphine palladium (15 mg, 0.013 mmol). The mixture was stirred at ca. 22°C under a nitrogen atmosphere for 2 h. After addition of anhydrous Et^O (7 mL), the precipitate was filtered, washed with anhydrous Et^O (7 mL) and dried Zn vacuo to yield 101 mg of brownish solid.' This was purified by reverse phase column chromatography (C, Q PrepPAK, 12 g, Waters Associates) eluting with H_0. lo 2 Appropriate fractions (fr. 7-12, each 20 mL) were collected and lyophilized to obtain 53 mg (0.16 mmol, y. 64¾) of the title compound as yellowish powder. This material was contaminated with potassium diphenylphosphate and potassium 2-cthylhexanoate: ’Hmr <0^0, CFT-20) ¢: 0.80 (t, £=6.4 Hz, Me from ethylhexanoate) , 1.21 (3H, d', £«6.3 Hz, l‘-Me), 2.93 (2H, dd, J. *9 Hz, J =4 Hz, 1-Hs), 3.23 (1H, dd, J. ,,=6.2 Hz. —1~5 —gem —o-i J, = 2.5 Hz, 6-H), 3.42 (2H, t, J"6 Hz, -CH S), 3.98 (1H, td, J. —o-5 " 2 ’'□I Hi, £, ,- = 2.5 Hz, 5-H), 4.15 (lH, q, J=6.2 Hz, I'-H), 4.80 Ϊ2Η, t, £"6.0 Hz, -*CH2N+) t 7-7.5 (m, phenyl protons from diphenyl phosphate), 8.03 (2h, m, Hm of pyridinium), S.56 (1H, m, Hp of pyridinium) and 8*81 ppm (2H, d, £=6.5 Hz, Ho of pyridinium) .

Example lg Preparationof 3-(2-(N-Methyl-thiomorpholinium)ethylthio]6a-[1'-(R)-hydroxyethyl)-7-oxo-l-azabicyclo(3.2. 0]-hept2-ene-2-carboxylate A. N-methy1-N- (2-mercaptoethyl)thiomorpholinium methanesulfonate «θ 1) Ms OH 2>Δ MsO To precooled (ice bath) N-methy1thiomorpholine" (5.00 g, 42.7 mmol) was added methanesulfonic acid (1.47 mL, .5 mmol) and ethylene sulfide (1.30 mL, 21.4 mmol). The mixture was heated at 65°C for 24 h and diluted with water (25 mL). The aqueous solution was washed with diethyl ether (3 x 25 mL), pumped under vacuum and poured over a silica gel reverse phase column; the title compound being eluted with J 5 water. The appropriate fractions were combined and evaporated to afford the thiol as an oil ¢4.80 g, yield 86%); ir (film) υ : 2550 cm-1 (w, SH) ; Simr (DWSO d,)5: 3.25-2.95 (6H, m, max ¢. b CH2N ) , 3.32 <3H, s, CH^N ) , 3.20-2.65 (7H, m, CH2S, SH) and 2.32 ppm (3H, s, CH3SO3).

*J.M. Lehn and J. Wagner. Tetrahedron, 26 , 4227 (1970) para-Nitrobenzyl 3-[2-(N-methyl-thiomorpholinium diphenyl phosphate) ethy1thio]-6a - [11 - (R)-hydroxyethyl)-7-oxo-lazabicyclo [3. 2.0]-hept- 2-ene-2-carboxylate OH 1) 0, 2). C1PO(O$)3 OH C0,PN3 4) EtN(—<)2 C02?N3 ^(Οφ)2 A cold (ice bath) solution of para-nitrobenzyl 6α- 11'- (R)-hydroxyethyl]-3,7-dioxo-l-azabicyclo(3.2.0)heptane2-carboxylate (557 mg, 1.60 mmol) in CH^CN (8 mL) was treated dropwise with diisopropylethyl amine (0.336 mL, 1.92 mmol) and diphenylchlorophosphate (0.400 mL, 1.92 mmol) and stirred for min. The reaction mixture was treated with N-methy1-N-(2mercaptoethyl)thiomorpholiniurn methanesulfonate (993 mg, 2.29 mmol) in CH^CN (4 mL) and diisopropylethyl amine (0.336 mL, 1.92 mmol) and stirred for 30 min. The solution was diluted with water (20 mL) and poured over a silica gel reversed phase I 5 column. The desired compound was eluted with a 50¾ acetonitrilewater mixture. The appropriate fractions were combined, pumped under vacuum for 2 h. and lyophylized to afford the title compound (1.01 g, yield 85%): ir (nujol)v : 1760 (s, S-lactam C=O) and 1510 cm1 (s, NO2) ; Hmr (DMSO-dg) 6: 8.25 (2H, d, J=8.8H2, H-aromatic), 7.70 (2H, d, J=8.8 Hz, H-aromatic ) , 7.336.84 (10 H, m, H-aromatic), 5.37 (2H, center of ABq, J=14.2 Hz, CH2) , 5.14 (1H, d, J=4.5 Hz, OH), 4. 35-3.80, (2H, m, H-l* and H-5), 3.75-3- 45 (6H, m, CH2N*) , 3.31 (3H, s, CH3N+) , 3.45-2.75 (9H, m, CH2S, H-6 and H-4) and 1.15 ppm (3H, d, £=6.2 Hz, CH^)·. a C. 3-[2-(N-methy1-thiomorpholinium) ethylthio]-6α-[1’- (R)hydroxyethyl]-7^oxo-l-azabicyclo(3.2.0]-hept-2-ene-2- carboxylate «ϊ • OH ’f r> A solution of para-nitrobenzyl 3-[2-(N-methylthiomorpholinium diphenylphosphate)ethylthio]-6a- [1’-(R)hydroxyethyl]-7-oxo-l-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (1.31 g, 1.76 mmol) in 0.1M pH 7.4 phosphate buffer (48.8 mL), tetrahydrofuran (20 mL) and diethyl ether (20 mL) was hydrogenated over 10% pd/C (1.5 g) in a Parr shaker for 1 h at 40 psi. The reaction mixture was diluted with diethyl ether (40 mL) and the phases were separated. The organic phase was washed with water (2x5 mL). The aqueous phases were combined, filtered through a #52 hardened filter paper, washed'with diethyl ether (2 x 20 mL) and pumped under vacuum. The aqueous solution was poured on a silica gel reverse phase column and the desired carbapenem was eluted with 5% acetonitrile-water. The appropriate fractions were combined, and lyophilized to give title compound as an amorphous solid (205 mg, 31%); ir (nujol)v : 1750 (s, β-lactam C=O) and 1590 cm-1 (s, C-O); Hmr (D2O) 0:4.25-3.95 (2H, m, H-l', H-5) , 3.70-3.40 ( 6H , m, CH2N*) , 3.35 (1H, dd, J=6.1 Hz, J=2.6 Hz, H-6), 3.08 (3H, s, CH.jN+) , 3.25-2.75 (8H, CHjS, H-4), and 1.24 ppra <3H, d, J=6.4 Hz, CH,); uv (H,0, c 0.062)λ„: 299 (ε1Ο,962) Tl/2 17.7 h (0.1M pH 7 phosphate buffer, 37°C) . 3G Example 17 Preparation of (5R, 6S)-3- [2-(1-methylmorpholino)ethylthio]6- [ (R) -l-hydroxyethyl)-T-oxo-l-a2abicycro’(’3.'2’."0]"hept-2-ene2 - car boxy l~a te l-.Methyl-l- (2-mercaptqethyl)morpholinium trifluoromethane- To N-methy lmorpholine (3.29 mL/ 0.030 mol) was added W dropwise trifluoromethanesulfonic acid (1.327 mL, 0.015 mol) at 10°C, followed by ethylene sulfide (0.89 mL, 0.015 mol). The resulting yellow-brown solution was heated (oil bath) at 50-60°C under for 13 h. Volatile material was then removed in vacuo and the residual oil was taken up in 10 mL of The aqueous solution was washed with diethyl ether (3x5 mL) and then residual organic solvent was removed in vacuo. The resulting aqueous solution was applied to a C^g reverse-phase column which was eluted with H^O, then 5% acetonitrile-h^O and finally 10¾ acetonitrile-F^O- Evaporation of the relevant fractions afforded a white solid which was dried in vacuo (P2O5) to give the product (1.92 g, 41¾). ir (KBr)v : 2560 (-SH) cm \· ^Hnmr (d^-acetone) «·. 4.25-3.6 (m, 8H) , 3.49 (s, 3H, N-Me), 3.35-2.7 (m, 5H) .

In py *J < B. p-Nitrobenzyl (5R,6S)-3-(2-(1-methylmorpholino)ethylthio)- ’ 6- ( (R) -l-hydroxye’thy 1) - 7-oxo-1-a'zab I cyclo[3.2.0] hep't-2-ene2-carboxylate diphenylphosphate To a solution of p-nitrobenzyl (5R,6S)-6-[(R)-1hydroxyethyl]-3,7-dioxo-l-azabicyclo[3.2.0]heptane-2-carboxylate (0.348 g, 1.0 mmol) in 25 mL of dry acetonitrile was added dropwise diisopropylethylamine (0,191 mL, 1.1 mmol) and then diphenyl chlorophosphate (0.228 mL, 1.1 mmol) at 0°C under After stirring at 0°C for 1 h diisopropylethylamine (0.226 mL, 1.3 mmol) was added to the resulting enol phosphate, followed by 1-methyl1-(2-mercaptoethyl)morpholinium trifluoromethanesulfonate (0.373 g 1.2 mmol). The reaction mixture was stirred at room temperature for 1.5 h and then concentrated in vacuo. The residual material was taken up in H^O and applied to a C^g reverse-phase column.

Elution with then 20% acetonitrile-H^O and finally 30% acetonitrile-H^O followed by lyophilization of the relevant fractions gave the product ¢0.360 g, 40%) as an amorphous solid. ir (film) 3300 (-OH), 1770 (β-lactam CO), 1700 (-CO2PN3) cm1; 1Hnmr (dc-acetone) 6: 8.25, 7.80 (ABq, J=8.6 Hz, 4H, aromatic), o 7.4-6.8 (m, 10H, diphenylphosphate), 5.56, 5.27 (ABq, J=14.2 Hz, 2H, benzylic), 4.42 (d of t, J=9.2 Hz, J'=2.7 Hz, IH, H-S), 4.12.7 (m, 17H), 3.40 (s, 3H, N-Me) , 1.22 (d, J=6.2 Hz, 3H, -CHMe).

C. (5?., 65) - 3-[2- (l-methylmorphQli.no) ethyl thio] -6 - [ (R) -1h y d ro xv e thyl] - 7-oxo-1-a zab i cv clo[3.2.0}h ep t-2-e ne -2 carboxylate To a solution of p-nitrobenzyl (5R,65) -3-[2-(1methylmorpholino)ethylthio]-6-[(R)-1-hydroxyethyl]-7-oxo-lazabicyclo [3.2.0]hept-2-ene-2-carboxylate diphenylphosphate (0.360 g, 0.49 mmol) in 13 mL of phosphate buffer (0.05 M pH 7.4) was added 0.36 g of 10% palladium-on-charcoal, 20 mL of tetrahydrofuran and 20 mL of diethyl ether. This mixture was hydrogenated (Parr) at 32 psi for 1 h. The mixture was filtered through Celite and the filter pad was washed with H^O and diethyl ether. The aqueous phase was separated and the pH was adjusted to 7.0 with additional pH 7.4 phosphate buffer. )5 After removing residual organic solvents in vacuo the aqueous solution was applied to a C^g reverse-phase column. Elution with H^O and lyophilization of the relevant fractions afforded 0.130 g of an amorphous solid. This material was repurified by reverse-phase hplc to give the pure product (0.Q5S g, 34%) 20 as an amorphous solid. ir (KBr)v : 3420 (br, OH), 1750 • . max (8-lactam CO), 1590 (-CO2") cm ; Hnmr (DjO) 6: 4.35-2.77 (m, 17H), 3.18 (s, 3H, N-Me) , 1.23 (d, J=6.3 Hz, 3H, CHMe); uv (Η-Ο)λ : 300 (eS344) nm; t, ,, (pH 7.4, 36.8^0 18.5 h. 2, max x/ z Examole 18 Preparation of (5R,65)3-[2-(1,4-dimethyl-l-piperazinium) ethy1thio]-6- [1- (R)-hydroxyethy1]-7-oxo-i^azabicyclo(5.2.0Thept-2-ene-2-carboxvlate I-(2-acetyithioethy1)-1,4-dimethylpiperazinium bromide ’N N· v_/ Acetone 50°C A solution of 2-bromoethyl thiolacetate *(2.20 g, 0.012 mol) and 1,4-dimethylpiperazine ¢1.95 mL, 0.014 mol) in acetone (4 mL) was stirred at 50®C for 65 h. After cooling to 25°C, the liquid phase was decanted from the gum which was triturated twice in diethyl ether; a hygroscopic yellowish powder, 3.2 g (90%) was obtained; ir (Nujol) v : 1685 (OO of thioester) cm ; ^Hmr (D20) 6; 2.37, 2.39 (2s, 6H, CH ^0, 2>-CH3), 3.18 (s, 3H, 2/N^3 ).

*B. Hansen, Acta Chem. Scand. 11, 537-40 (1957) to B. 1,4-dirnethyl-l-(2-mercaptoe thy 1) piperazinium bromide hydrochloride HC1 ‘Hmr (DMSO, άθ) A solution of 1-(2-acetylthioethy1)-1, 4-dimethyl5 piperazinium bromide {1.1 g, 3.7 mmol) in 6N hydrochloric acid (4 mL) was heated at 80°C under a nitrogen atmosphere for 1 h. The solution was concentrated under reduced pressure to give δ: 2.90 (s, 7.16, N 9.05, 19. a white powder, 0.41 g (38%), )/ >, 3.26 (3,3/33 —7 \ch3 —7 Anal. calcd. for CgH20N2SBrCl· H^O: C 31.03, H S 10.35; found: C 31.62, H 7.46, N 9.19, S 10.

C. (5R,6S)Paranitrobenzyl 3-(2-(1,4-dimethyl-l-piperazinium)ethylthio]-6-[1-(R)-hydroxyethyl]-7-oxo-l-azabicyclo(3.2.0Jhept-2-ene-2-carboxylate diphenylphosphate To a cold (0°C) solution of (5R,6S) paranitrobenzyl 6-(1 - (R)-hydroxyethyl]-3,7-dioxo-l-azabicyclo[3.2.0]heptane-2(R)-carboxy late (0.465 g, 1.33 mmol) in acetonitrile (2 mL) kept under a nitrogen atmosphere was added diisopropylethylamine (0.278 mL, 1.59 mmol) and diphenyl chlorophosphate (0.33 mL, 1.59 mmol). The reaction mixture was stirred for 30 min and treated with a suspension of 1,4-dimethy1-1-(2-mercaptoethy1)piperazinium bromide hydrochloride (0.40 g, 1.37 mmol) in acetonitrile (3 mL)-water (1 mL) mixture and diisopropylethylamine (0.278 mL, 1.59 mmol). After stirring for 18 h at 5°C, cold water (15 mL) was added to the mixture. The resulting solution was chromatographed over PrepPak-500/C^g (Waters Associates) column (2.5 x 7.5 cm) with 25-35% acetonitrile in water as eluting solvents to give a yellowish powder 0.50 g (50%) after : 1765 (C=O of β-lactam), 1690 lyophylization; ir (KBr) v^ (C=O of PN3 ester), 1585 (phenyl)·, 1512 (NO2) , 875 (N0£) XHmr (DMSO, dg) δ: 1.16, 1.18 (2d, J=6.1 Hz, 3H, CHgCHOH) , 2.44 (s, cm N CH3) , 3.14 (s, ), 5.31 (a, j=6 Hz, OK), 5.39 (center of ABq, J=13 Hz, CH2 of PNB), 6.6-7.4 (m, 10H, phenyl of phosphate), 7.71 (d, >8.8 Hz, 2H, Ho of PNB), 8.26 (d, >8.8 Hz, Hm of PNB) . (5R,6S)3-(2-(1,4-dimethy1-L-piperazinium)ethy1thio]-6* [ 1- (R)-hydroxyethyl]-7-oxo-l-azabicyclo[3.2.0]hept-2ene-2-carboxylate OH %, Pd/C > THF, Etherf bufCar OH r coo To a solution of ί5R,6S)paranitrobenzyl 3-[2-(1,4dimethyl-l-piperazinium)ethylthio)-6-[1-(R)-hydroxyethyl]-7oxo-l-azabicyclo[3.2.0]hept-2-ene-2-carboxylate diphenylphosphate (0.47 g, 0.623 mmol) in wet tetrahydrofuran (25 mL) was added diethyl ether (25 mL), potassium phosphate monobasic-sodium hydroxide buffer (13 mL, pH 7.22) and 10% palladium on charcoal (0.47 g). The resulting mixture was hydrogenated at 23eC under 40 psi for 1 h. The two layers were separated and the organic layer was extracted with water (2x7 mL). The aqueous layers were combined, filtered through a Celite pad, washed with diethvl15 ether (2 x 15 mL) and chromatographed on PrepPak-500/C^g (Waters Associates) column (2.5 x 9.5 cm) with water as eluting solvent to give, 0.097 g (43%) after lyophylization; ir (KBr) v : max . 3000-3700 (OH), 1750 (C=0 of β-lactam), 1585 (carboxylate)cm , ^mr (D^O) δ: 1.24 (d, J=6.4Hz, 3H, CH^CHOH) , 2.33 (s, 3H, (measured at a concentration of 10 at 36.8eC).

Example 19 Preparation of (5R, 6S) -3- [2-- (N-methy1-thiomorpholiniumoxide)ethylthio]-6-Ϊ1-(R)-hydroxyethyl]-7-oxo-l-azabicy clo(3.2.0)-hept-2-ene-2-carboxylate mCPBA ------y- To a cold (-10QC) solution of (5R,6S)-3[2- (N-methy 1-thiomorpholiniuin) ethylthio] -6- [1- (R) -hydroxyethyl] 7-oxo-1-azabicyclo{3.2.0)-hept-2-ene-2-carboxylate (608 mg, 1.65 mmol) in a 1:1 mixture of acetonitrile-water (9 mL) was added m-chloroperbenzoic acid (334.8 mg, 1.65 mmol) in small portion over a 1 hour period. The mixture was then diluted with water (15 mL) and washed with diethyl ether (3 x 15 mL) .

The aqueous phase was pumped under vacuum and passed through a reversed phase silica gel column (^0) to give a solid which consisted of a mixture of compounds. This mixture was separated by reversed phase HPLC and afforded fraction A 52.4 mg (yield 12%) and fraction B 23.6 mg (yield 6%) as diastereomers of the title compound; Fraction A: ir (nujol) v : 1750 (s, S-lactam C=0) . a max and 1580 cm_J· (s, C=0) ; Hmr (DjO) δ: 4.26-2.91 (20 H, m, H-4, H-5, H-6, H-l', CH2S, CH2S-O, CH^-N* and CH2N+) and 1.24 ppm ( 3H , d, J=6.4 Hz, CH,) ; uv (H,0, c 0.06) λ„ : 302 (el0425); j 2 max T 1/2:12 h (0.065 M, pH 7.4 phosphate buffer, 37°c). Fraction 3: ir (nujol) v : 1750 (s, β-lactam C=O) and 1585 cm 1 (s, C=O); •"Hmr (D20) δ: 3. 86-2.90 (17 H, m, H-4, H-5, H-6, H-l', CH2S, CH2S-O, CH2N+) , 3.25 (3H, s, CH^N4) and 1.24 ppm (3H, d, J=6.4 Hz, 1'< CH,); uv (H,0, c 0.05) λ : 299 (e6517) ; T 1/2:10.75 h (0.065 M, 2 max pH 7.4 buffer solution, 37°C).

Example 20 Preparation of (5R,6S)-3-[2-(1,4,4-Trimethy1-1-piperazinium)ethylthio]-6-(lR-hydroxyethy1]-7-oxo-1-azabicyclo(T. 2.0)hept2-ene-2-carboxylate chloride A. 1-(2-acetylthioethyl) -1,4, 4-trimethylpiperazinium bromideiodice Sr KeX OH, 55-60®C ' A suspension of 1-(2-acetylthioethyl)-1,4-dimethylpiperazinium bromide (1.48 g, 5.0 mmol) in isopropyl alcohol (10 mL) was treated with methyliodide (0.373 mL, 6.0 mmol) and heated at 55-60°C for 30 h. The solvents were evaporated under reduced pressure the residue was triturated in hexane and the solid was filtered, 1.85 g. The solid was dissolved in hot water (8 mL) and the solution was diluted with acetone until turbidity (70-80 mL). Two successive crystallizations gave 1.5 g, mp 220-5°C dec., 68¾ of the title compound; ir (K3r)v : 1692 cm 1 (C=0); ^Hmr (D-.0) 6: 2.40 (s, 3H, CH3COO) , 3.37 (s, N-CH3) , 3.39 (s, N-CH3) , 3.99 (s) ; uv (I^OJX^: 226 (el3144). Anal, calcd for ^OSBrl: C 30.08, H 5.51, N 6.38; found: C 30.48, H 5.53, N 6.86. 3. 1- (2-mercaptoethyl) -1,4, 4-trimethvlpiperaziniun bischloride Permutit S-l cr_ Cl A mixture of 1-( 2-acctylthiocthyl. ) - 1,4,4-trimethylpiperazinium bromideiodide (1.84 g, 4.19 mmol) and 6N hydrochloric acid (15 mL) was heated at 57°C under a nitrogen atmosphere for 2.5 h. The solution was concentrated under reduced pressure to dryness- The solid was suspended in water (10 mL) and the wellstirred suspension was treated with permutit S-l Cl* until a solution was obtained. The solution was pour.ed on a column’(1.2 x cm) of permutit S-l Cl . The column was eluted with water (1.5 mL/min). The appropriate fractions were combined and lyophilized to give a white powder, 0.93 g, mo 190-191°C, 85%; ir (nujol)\i : * * max 2450 (SH) ; Aifcnr (D O) δ: 3.4 (s, N-CH-j) , 3.45 (s, N-CHp , 4.07 (s). Anal, calcd for C9H22N2SC12‘0.75 H2O: C 39.34, H 8.62, N 10.20, S 11.67; found: C 39.48, H 8.39, N 10.55, S 11.15. 147 C . (5R,6S) parani trobenzyl 3-(2-(1,4,4- trimethyl-1-piper a zinivm) ~ ethy1 thio f - 6 - (ί R-hyc r oxyethy1 ] - 7 -οχο-Ύ-a zab i cyclo (3.2.0)hept2-ene-2-carboxvlate bischloride 1) NEt(i?r)2 2) Ci?(O?h)j 3) N2t(i?r) Ό00ΡΝ3 4) ?() To a cold (5°C) solution of (5R,6S) paranitrobenzyl 6[IR-hydroxyethy1]- 3,7-dioxo-l-azabicyclo(3.2.0)heptane-2R-carboxy late (0.94 g, 2.7 mmol) in acetonitrile (3 mL) kept under a nitrogen atmosphere was added diisopropylethylamine (0.557 mL, 3.2 mmol) and diphenyl chlorophosphate (0.663 mL, 3.2 mmol). The reaction mixture was stirred at 5°C for 30 min and treated with diisopropylethylamine (0.599 mL, 3.44 mmol) and an aqueous solution (4 mL) of 1-(2mercaptoethyl)-1,4,4-trimethyloiperazinium bischloride (0.90 g, 3.44 mmol). After 1.25 h, diisopropylethylamine (0.1 mL, 0.57 mmol) was added and the stirring was continued for 2 h. A part of the acetonitrile was eliminated under reduced pressure and the resulting red mixture was chromatographed on PreoPak -500/C,A (Water Associates) column with 25-75% acetonitrile in water as eluting solvent to give a yellowish powder (1.4 g) after lyophylization. The powder was solubilized in water and the solution was passed on a column (1.2 x 58 cm) of permutit S-l Cl using water an eluting solvent. Lyophvlization of the appropriate fractions gave 1.17 g of a powder that was reourified on a column of PreoPak -500/C Lyoohvlization of the appropriate fractions gave a yellowish powder, 0.80 g (53%); 3400 (br, OH), 1770 (C=0 of the £-lactam) , 1690 ir (X3r)\j max 2S 1605 (aromatic), 1515 (NOJ , 134 5 (NO J (C=0 of PN3 ester 1Hmr (D20) c: 1.26 (d, >6.3 Hz, 3H, CF^CHOH) , 3.39 (s, NCH3) , 4.00 (s) .37 (br, s, CH2 of PN3) , 7.60 (d, >8.6 Hz, 2H UV (H20)Amax Anal, calcd. for C^cH^-N.O.SCl·’2H-0 25364ο 2 2 JO 8.20 (d, >3.7 Hz, 2H, Hm of ?N3) (: 10 752) .

N- 3.63, £ 4.97, Cl 10.93; found; C Cl 11.37.

Ho of· PN3) , 276 (ε12094), 306 C 46.51, H 6.56, ' <1 8 D. (5R, 6S) -3-(2-(1/4, 4-trimethy1- 1-piperazinium) ethylthio] -6(IR-hydroxyethy 1]-7-oxo-l-azabicyclo(3.2.0)hept~2-ene-2carboxvlate chloride A mixture of (5R,6S) paranitrobenzyl 3-(2-(1,4,4trimethy1-1-piperazinium)ethylthio] -6-(lR-hvdroxyethyl]-7-oxo-1a2abicyclo(3.2.0)hept-2-ene-2-carboxylate bischloride (0.40 g, 0.63 mmol), phosphate buffer (30 mL, 0.05M, pH 7.0), tetrahvdrofuran (10 mL), ether (30 mL) and 10% palladium on charcoal (0.40 g) was hydrogenated at 23°C under 35 psi for 1 h. The two phases were separated. The organic phase was extracted with water (10 mL).

The aqueous phases were filtered on a Celite pad, washed with ether . (10 mL) , concentrated under vacuum to 10 mL and chromatographed cr.

Pre?ak-500/C^g column (2.2 x 11 cm) with water as eluting solvent I '> to give 70 mg (25%) after lyophylization; ir (KBr)v : 3400 (br, OH), -11¼ 1755 (C=O of the 5-lactam), 15S5 (carboxylate) cm ; Kmr (D^O) c* : 1.24 ( 3H , d, J = 6.3 Hz, CH^CHOH), 3.36 (s, NCH^) , 3.93 (s) ; uv (I^O) Xmax: 296 <ε 7997)'· [°3ο3 35’9 10° (c' °·30' H20) ' Tl/2= * * *9'S h Measured at a concentration of ϊθ4 M in phosphate buffer pH 7.4 at 36.8°C) .

Claims (18)

1. A process for the preparation of a carbapenem derivative of the formula S-A-R COOR 2 S wherein R S * * 8 * 10 * * * * 15 is hydrogen and P 1 is selected from hydroacn; substituted and unsubstituted; alkyl, alkenyl and alkynyl, having from 1-10 carbon atoms; cycloalkyl and cycloalkylalkyl, having 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and 10 aralkynyl wherein the aryl moiety is phenyl and the aliphatic portion has 1-6 carbon atoms? heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl wherein the hetero atom or atoms in the above-named heterocyclic moieties are selected from J-4 oxygen, nitrogen or sulfur atoms and the 15 alkyl moieties associated with said heterocyclic moieties have 1-6 carbon atoms; wherein.the substituent or substituents relative to the above-named radicals are selected from ISO C,-C, alkvl optionally substituted bv lb amino, halo, hydroxy or carboxyl halo -OR 3 ll 34 -OCNR R ll 34 -CNR R 3 4 -NR λ it 3 4 -SO 2 NR R 0 N 34 -NHCNR R 3/1 4 R CNR -c° 2 r3 =0 -OCR 3 -SR 3 // 9 -SR n 9 -SR · II -CN -N„ -0S0 3 R -OSO o R 3 3^ 4 -NR SO.R 3 ^4 -NRC=NR 3 4 -NR CO 2 R -NOwherein relative to the above-named substituents, the groups R and R are independently selected from hydrogen; alkyl, alkenyl and alkynyl, having from 1-10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalky1, having 3-6 carbon atoms 5 in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl wherein the aryl moiety is phenyl and the aliphatic portion has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl § and heterocyclylalkyl wherein the hetero atom or atoms in the 10 above-named heterocyclic moieties are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with said heterocyclic moieties have 3 4 1-6 carbon atoms, or R and R taken together with the nitrogen to which at least one is attached may form a 5- or 6- membered 9 3. l5 nitrogen-containing heterocyclic ring; R is as defined for R 1 8 except that it may not be hydrogen; or wherein R and R taken together represent alkylidene or C 2~ C 10 alkylidene substituted by hydroxy; A is cyclopentylene, cyclohexylene or C_-C alkylene optionally substituted by one or more C.-C* 2 A 4 20 alkyl groups; R is hydrogen, an anionic charge or a readily removable carboxyl protecting group, providing that when R is hydrogen or a protecting group, there is also present a counter anion; and R is a guaternized nitrogen-containing aromatic or non-aromatic heterocycle attached 25 to A through a ring nitrogen, thereby forming a quaternary ammonium group, or a pharmaceutically acceptable salt, thereof, which process comprises reacting an intermediate of the formula wherein readily leaving L COOR 2 ' 18 2 1 R and R are as defined above, R removable carboxyl protecting group group, with a thiol compound of the HS-A-R 14 VII rf is a and L is a formula 14 Θ wherein A and R are as defined above and X is a counter anion, in an inert solvent and in the presence of base,to produce a carbapenem product of the formula 182’ 14 Θ wherein R , R , R , A, R and X are as defined above and, 2’ if desired, removing the carboxyl protecting group R to give the corresponding de-blocked compound of formula I, or a pharmaceutically acceptable salt thereof·>.

2. Λ process according to Claim I,wherein the base is a non-nucleophilic tertiary amine or a tri(C^-C^) alkylamine .

3. Λ process according to Claim 2,wherein the base 5 is a non-nucleophilic tertiary amine.

4. A process according to Claim 1, 2 or 3,wherein the reaction is carried out at a temperature of -15°C up to room temperature.

5. Λ process according to claim 4, wherein the 10 reaction is carried out at a temperature in the range of 1'rom -15°C to i l ')°t;.

6. A process according to Claim 1, 2, 3, 4 or 5, wherein the inert solvent is selected from acetonitrile, a mixture of acetonitrile, dimethylL5 formamide, tetrahydrofuran, a mixture of tetrahydrofuran and water, a mixture of acetonitrile and water and acetone.

7. A process for the preparation of a quaternary amine thiol compound of the formula HS — A —R 14 154 wherein A is cyclopentylene, cyclohexylene or R in which R , R , R and R are each independently hydrogen Θ 14 or C^-C^ alkyl, X is a counter anion and R represents a substituted or unsubstituted mono, bi- or polycyclic aromatic or non-aromatic heterocyclic radical containing at least one nitrogen in the ring and attached to A through a ring nitrogen, thereby forming a quaternary ammonium group, which process comprises reacting a sulfide of the formula or wherein R^°, R^\ R^ and are each independently hydrogen or alkyl with a strong acid and either (a) a heteroaromatic amine of the formula represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocyclic radical containing at least one nitrogen in the ring capable of being guaternized and bonded to a carbon atom 10 of substituent A or (b) a heterocyclic amine of the formula represents a substituted or unsubstituted mono-, bi- or polycyclic l5 non-aromatic heterocyclic radical containing at least one nitrogen in the ring capable of being guaternized by substituent R 1 ^ and 1 β bonded to a carbon atom of substituent A and R represents either (a) an optionally substituted aliphatic, cycloaliphatic, cycloaliphatic aliphatic, aryl, araliphatic, heteroaryl, heteroaraliphatic, hetero20 cyclyl or heterocyclyl-aliphatic radical or (b) a divalent phenylene or C^-C^ alkylene group joined to the ring so as to form a bridged polycyclic group.

8. A process according to Claim 7, wherein the reaction is carried out at a temperature in the range of from -20°C tolOtfc.

9. A process according to Claim 7 or Claim G, wherein the si. ronn acid is hydrochloric, hydrobromic, methanesulfonic, ptoluenesul fonic or trifluoromethancsulfonic. Ϊ ΙΟ. Λ process according to Claim 7, 8 or 9, wherein the reaction is carried out in the presence of a non-polar organic solvent. Ί ll. A process according to Claim 10, wherein the non-po.l.ar organic solvent is methylene chloride, benzene, xylene or toluene.

10. 12. A process according to Claim 7, 8, 9, 10 or ll, wherein, when the amine and sulfide reagents are liquids or when the amine reagent is a solid, the sulfide reagent is liquid and the solid amine in soluble in the liquid sulfide reagent, the reaction i:-. carried out without additional solvent. I r > 13. A quaternary amine thiol compound of the formula HS — A — R χθ VII wherein A is cyclopentylene, cyclohexylene or R 10 R 12 I I —c-c — i 11 i 13 in which R^^, κΔ R^ and R^ are each independently hydrogen 20 or C^-C^ alkyl, Χθ is a counter anion and R^ represents a substituted or unsubstituted mono, bi- or polycyclic aromatic or non-aromatic heterocyclic radical containing at least one nitrogen in the ring and attached to A through a ring nitrogen, thereby forming a quaternary ammonium group. I Ify ρ* Γί <

11. 14. A process according to Claim 1, substantially as described in any of the applicable Examples in the foregoing description.

12. 15. A process according to Claim 7 t substantially as 5 described in any of the applicable Examples in the foregoing description.

13. 16. A process according to Claim 1, modified by the inclusion of an additional step, in which process '* 14 j? R comprises initially an N-methyl-thiomorpholinium 10 radical, and the initially obtained derivative of formula I is converted into the corresponding compound answering 14 to formula I wherein R comprises an N-methylthiomorpholinium oxide radical, by oxidation with m-chloroperbenzoic acid, the procedure being that of the * r > foregoing Example 19 or an. r analogous procedure equivalent thereto.

14. 17. A carbapenem derivative prepared by a process according to any of claims 1 to 6 or Claim 14.

15. 18. A quaternary ammonium thiol compound prepared by a process according to any of claims 7 to 12 or Claim 15. 20

16. 19. A sulfoxide compound prepared by the process of Claim 16.

17. 20. A pharmaceutical composition comprising a carbapenem derivative as claimed in claim 17 and a pharmaceutically acceptable carrier or diluent.

18. 25 21. A pharmaceutical composition according to Claim 20, substantially as hereinbefore described.