DD237507A5 - PROCESS FOR THE PREPARATION OF QUATEROUS AMINTHIOL INTERMEDIATE COMPOUNDS - Google Patents

Abstract

The invention relates to a process for the preparation of quaternary Aminthiol.-intermediate compounds of the general formula VII HS-A-R14 XqVII wherein A is cyclopentylene, cyclohexylene or wherein R10, R11, R12 and R13 independently of one another is a hydrogen atom or a C1-4 alkyl group X, represents a counter anion and R 14 represents a substituted or unsubstituted, mono-, bi- or polycyclic, aromatic or non-aromatic, heterocyclic radical containing at least one nitrogen atom in the ring and attached to A via a ring nitrogen atom, thereby forming a quaternary ammonium group is formed. The intermediate compounds obtainable according to the invention can be further processed into carbapenem derivatives.

Description

Field of application of the invention.

The invention relates to a process for preparing quaternary Aminthiol intermediates, which can be further processed to carbapenem derivatives, which in the 2-position Substiuenten the formula

-SAR ¹⁴

in which A is a cyclopentylene, cyclohexylene or C 2-6 -alkylene radical which is optionally substituted by one or more C 1-4 -alkyl groups, and R _{14 is} an aromatic or non-aromatic heterocycle which contains a quaternized nitrogen atom and is bonded to A via a quaternary nitrogen atom is bound.

Characteristic of the known technical solutions

The carbapenem derivatives prepared with the intermediates obtainable according to the invention are described in US patent applications Ser. 366,910,471,379 and 389,652 (invent .: Choung U. Kim and Peter F. Misco Jr.). The entire disclosure of each of these applications is hereby incorporated by reference.

US Application 366 910 and its continuation application, Serial No. 471 379, which corresponds to DE-OS 3 313 533, describe the preparation of carbapenem antibiotics of the formula

IA ^v .

R ^{8 represents} a hydrogen atom and

R ^{1 represents} a hydrogen atom or represents the following substituted and unsubstituted radicals:

an alkyl, alkenyl and alkynyl group having 1 to 10 carbon atoms; a cycloalkyl and cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkylation units; a phenyl group; an aralkyl, aralkenyl and araikinyl group wherein the aryl inte is a phenyl group and the aliphatic portion has from 1 to 6 carbon atoms; a heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl group, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen and sulfur atoms and the alkyl moieties attached to the heterocyclic moieties have 1 to 6 carbon atoms ; where the substituent or the substituents of the abovementioned radicals is one of the following groups:

a C _1-8 -alkyl group, which is optionally substituted by amino, halogen, hydroxy or carboxyl,

a halogen atom,

0 -OR ³ , -OCNR ³ R ⁴ , -CNR ³ R ⁴ Z-NR ³ R ⁴ , -

0 0 NR ³

^X NR ³ R ⁴

0 0

₂ R, -NHCNR ^ R, R ^ CNR, -CO ₂ R ^, = 0, 0 0 0 -OCR ³ , -SR ^, -SR ^y , -SR = *, -CN, -N ,, - OSO ^ R ^

0 -OSO ₅ R ³ , -NR ³ SO ₄ R ⁴ , -NR ³ C = NR ⁴ , -NR ³ CO ₉ R ⁴ or

NO ₂ ,

wherein, in the above-mentioned substituents, the groups R ³ and R "independently represent a hydrogen atom, an alkyl, alkenyl and alkynyl group having 1 to 10 carbon atoms, a cycloalkyl, cycloalkyl-acyl and alkylcycloalkyl group having 3 to 6 carbon atoms in the cycloalkyl ring and 1 a phenyl group; an aralkyl, aralkenyl and aralkinyl group, wherein the aryl moiety is a phenyl group and the aliphatic part has from 1 to 6 carbon atoms, or a heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl group, wherein the Heteroatom or the _Λ heteroatoms in the abovementioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moieties attached to said heterocyclic moieties have 1 to 6 carbon atoms, or R ³ and R ⁴ together with the Nitrogen atom to which at least one of these radicals is attached, e 5-membered or 6-membered nitrogen-containing heterocyclic ring;

R ^{9 is defined} as the group R ³ but can not denote a hydrogen atom;

or in which

R 'and R ⁸ together represent a C ₂ _i ₀ -alkyliden- or substituted by a hydroxy group C ₂ _ ₁₀ -Alkylidenrest; A represents a cyclopentylene, cyclohexylene or a C ₂ _e-alkylene group, if desired by one or more

Substituted C ^ alkyl groups;

R ^{2 represents} a hydrogen atom, an anionic charge or a common, easily removable carboxyl-protecting group, provided that when R ^{2 represents} a hydrogen atom or a protecting group, a counter anion is also present, and

the rest of the formula.

 Ό

represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocyclic radical containing at least one nitrogen atom in the ring and attached to A via a ring nitrogen atom, thereby forming a quaternary ammonium group;

and their pharmaceutically acceptable acid addition salts. These compounds are prepared by a method given in the following reaction scheme:

DiphenyIchlorphosphat

COOR

III

O OP (OC ₆ H ₅ )

COOR

HS-A-OH

A = alkylene or Cc-Cg -cycloalkylene

IV

methanesulfonyl

S -AOH

COOR

2 ¹

• Η

SA-OSO ₂ CH ₃

VI

'N

II

I ¹ A

COOR

2 ¹

S-A-I

COOR

₂ ,

© / -ν; -Α-Ν ϊ

COOR '

Ag ¹

= Counter anion)

 Ν

5-Α-Ϊ0

COOR

IA

US application 389,652 and continuation application 499,690 describe the preparation of carbapenem antibiotics of general formula IB

,1

R "

COOR ²

IB

wherein R ⁸ , R ¹ , R ³ , R * and R ^{9 have} the meanings given above;

R ^{5 is} selected from the following substituted and unsubstituted groups: alkyl, alkenyl and alkynyl of 1 to 10 carbon atoms; Cycloalkyl and cycloalkylalkyl having 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moieties; phenyl; Aralkyl, aralkenyl and aralkinyl, wherein the aryl moiety is a phenyl radical and the aliphatic moiety has from 1 to 6 carbon atoms; Heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl wherein the heteroatom or heteroatom in the above-mentioned heterocyclic moieties is selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moieties attached to said heterocyclic moieties have 1 to 6 carbon atoms; wherein the abovementioned R ⁵ radicals are optionally substituted by 1 to 3 substituents independently of one another

are selected from:

a C 1-4 alkyl radical which is optionally substituted by amino, fluoro, chloro, carboxyl, hydroxy or carbamoyl;

Fluorine, chlorine or bromine;

-OR ³ } -OCO ₂ R ³ ; -OCOR ³ ; -OCONR ³ R ⁴ ; -OSO ₂ R ³ ;

oxo; -NR ³ R ⁴ ; R ³ CONR ⁴ -; -NR ³ CO ₂ R ⁴ ;

0 -NR ³ CONR ³ R ⁴ J-NR ³ SO ₂ R ⁴ ; -SR ³ ; -SR ⁹ ;

-SR ⁹ ; -SO ₃ R ³ ; -CO ₂ R ³ ; -CONR ³ R ⁴ ; -CN; or

a phenyl radical which is optionally substituted by: 1-3-fluoro, chloro, bromo, C 1-4 -alkyl, -OR ³ , -NR ³ R ⁴ , -SO ₃ R ³ , -CO ₂ R ³ or -CONR ³ R *, wherein R ³ , R ⁴ and R ⁹ in these substituents R ^{6 have} the meanings given above; or wherein R ⁶ is a divalent phenylene or _C1 ^, - can mean alkylene group which with the ring _

is connected so as to form a bridged, polycycloic group;

A is a cyclopentylene or cyclohexylene radical or a C ₂ _e-alkylene group, if desired by an or

a plurality of C 1-4 -alkyl groups is substituted;

R ^{2 represents} a hydrogen atom, an anionic charge or a common, easily removable carboxyl-protecting group, provided that when R ^{2 represents} a hydrogen atom or a protecting group, a counterion is also present; and the rest of the

formula

a substituted or unsubstituted mono-, bi- or polycyclic non-aromatic heterocyclic radical containing at least one nitrogen atom in the ring and attached to A via a ring nitrogen atom, thereby forming a quaternary ammonium group; and the pharmaceutically acceptable salts thereof. This process is shown in the following reaction scheme:

COOR

diphenyl

III

IV.

O Il

OP (OC ₆ H ₅₎

COOR

HS-A-OH

A = alkylene or Cc-Cg-cycloalkylene

S-A-OH

methanesulfonyl

COOR

2 ¹

O'

SA-OSO ₂ CH ₃

Λ 2 '

COOR

VI

S-A-I

XOOR ²

Ao

I ¹ B

COOR

2 '

if necessary, the blockage

IB

S-A-N

COOR

In this known method, starting from the compound of formula III and sets them in an inert organic solvent with diphenylchlorophosphate in the presence of a base to the intermediate IV. Intermediate IV is then reacted with a mercaptan reagent of formula HS-A-OH in an inert organic solvent and in the presence of a base to give intermediate V. The intermediate compound V is then acylated with methanesulfonyl chloride in an inert organic solvent and in the presence of a base to give the intermediate compound VI which is reacted with an iodide ion source in an inert organic solvent to give the intermediate II. This intermediate compound II is reacted with the desired amine in an inert organic solvent and in the presence of silver ions to give the quaternized product l'A or ΓΒ, which can then be deblocked to yield the corresponding deblocked carbapenem compound of formula IA or IB ,

The method described above has several disadvantages. So this process goes through different stages, whereby it would be beneficial if you could reduce the number of stages. The yield of the overall reaction is also quite low. In addition, the quaternization is carried out with the entire carbapenem compound.

Object of the invention

By the method of the invention it is possible to obtain first the quaternary aminethiol intermediates, which are then bound to the carbapenem nucleus. The process for the preparation of the carbapenem derivatives therefore has fewer reaction steps and leads to higher yields. This also allows easier formation of quaternary amine products with a variety of amines, such as sterically hindered amines or those with low pK _b values.

Explanation of the essence of the invention

The object of the invention is to provide a process for the preparation of a quaternary aminothiol compound of the general formula VII

HS-AR * ⁴ Χ ^θ ' ^v11

wherein A is cyclopentylene, cyclohexylene or rio. R12

Il ρ r-

R ¹ U R13 - ,

wherein R ¹⁰ , R ", R ' ² and R ¹³ independently represent a hydrogen atom or a C ^ alkyl group, Χ ^{θ is} a Gegenanion and R ^{14 is} a substituted or unsubstituted, mono-, bi- or polycyclic, aromatic or is a non-aromatic heterocyclic radical which contains at least one nitrogen atom in the ring and is bonded to A via a ring nitrogen atom, forming a quaternary ammonium group The process according to the invention is characterized in that a sulphide of the general formulas

_R 10 SR ¹² or C- C

R ¹¹ R ¹ 3

wherein R ' ⁰ , R ", R' ² and R ' ³ each independently represent a hydrogen atom or a C 1-4 alkyl group, with a strong acid and either

(a) with a heteroaromatic amine of the formula «f \

this remainder

a substituted or unsubstituted, mono-, bi- or polycyclic, aromatic, heterocyclischeh radical containing at least one nitrogen atom in the ring, which can be quaternized and which is bonded to a carbon atom of the substituent A, or b) with a heterocyclic amine of formula

wherein the rest

a substituted or unsubstituted, mono-, bi- or polycyclic, non-aromatic, heterocyclic radical containing at least one nitrogen atom in the ring, which can be quaternized by the substituent R ¹⁶ and bonded to a carbon atom of the substituent A, and wherein R ¹⁶ either i) an optionally substituted, aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, heteroaraliphatic, heterocyclyl-aliphatic or araliphatic radical or an aryl, heteroaryl or heterocyclyl radical

or ii) a divalent phenylene or C, _4-alkylene group, which with the ring of the general formula

is connected so that a bridged, polycyclic group is formed, converts.

The reaction can be carried out in the presence or absence of an inert organic solvent. This solvent is preferably a nonpolar organic solvent such as methylene chloride, benzene, xylene, toluene or the like.

If the amine and sulfide reagents are liquids or if a solid amine is soluble in a liquid sulfide reagent, the reaction is preferably carried out without additional solvent.

The strong acid used for the reaction is not critical and may be, for example, a strong inorganic or organic acid such as hydrochloric acid, hydrobromic acid, methanesulfonic acid, p-toluenesulfonic acid, trifluoromethanesulfonic acid, etc.

The formation of the quaternary Aminthiol intermediate of formula VII can be carried out in a temperature range of about -20 ° to about 100 ° C. Preferably, one works at temperatures of about 50 to 70 ° C.

The sulfide reagent, the aromatic amine and the acid are preferably used so that the sulfide and acid are present in approximately equimolar amounts, with the amine being used in excess, e.g. B. 2 to 3 moles of amine per mole of sulfide or acid.

The quaternary Aminthiol intermediate has an associated counter anion derived from the acid used. It is of course possible, at this point, to replace the anion with another counter anion by conventional methods.

The Aminthiol intermediates obtainable according to the invention are further processed to give carbapenem derivatives of the general formula I:

,8th

S-A-R

COOR '

R ^{8 represents} a hydrogen atom and

R ^{1 represents} a hydrogen atom or represents the following substituted and unsubstituted radicals:

an alkyl, alkenyl and alkynyl group having 1 to 10 carbon atoms; a cycloalkyl and cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moieties; a phenyl group; an aralkyl, aralkyl and aralkinyl group wherein the aryl moiety is a phenyl group and the aliphatic moiety has from 1 to 6 carbon atoms; a heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl group, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen and sulfur atoms and the alkyl moieties attached to the heterocyclic moieties have 1 to δ carbon atoms ; where the substituent or the substituents of the abovementioned radicals is one of the following groups:

a C 1-4 alkyl group, which is optionally substituted by amino, halogen, hydroxy or carboxyl,

a halogen atom,.

-OR ^, -OCNR ^ R> -CNR ⁵ R, -NR ^, - //,

^X NR ³ R ⁴ OO

'r ⁴ , -NHCNR ³ R ⁴ , R ³ CNR ⁴ -, -CO ₂ R ³ , = O, OOO

-OCR- ³ , -SR- ⁵ , -SR 1, -SR ', -CN, -N _x , -OSO _x R 1,

πJ J

-OSOpR ³ , -NR ³ SO ₉ R ⁴ , -NR ³ C = NR ⁴ , -NR ³ COpR ⁴ or

wherein in the abovementioned substituents, the groups R ³ and R ⁴ independently of one another represent a hydrogen atom; an alkyl, alkenyl and alkynyl group having 1 to 10 carbon atoms; a cycloalkyl, cycloalkylalkyl and alkylcycloalkyl group having 3 to 6 carbon atoms in the cycloalkyl ring and having 1 to 6 carbon atoms in the alkyl moieties; a phenyl group; an aralkyl, aralkenyl and aralkinyl group wherein the aryl moiety is a phenyl group and the aliphatic part has 1 to 6 carbon atoms; or a heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl group, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moieties attached to said heterocyclic moieties have 1 to 6 carbon atoms have, stand, or

R ³ and R ⁴ together with the nitrogen atom to which at least one of these radicals is bonded form a 5- or 6-membered nitrogen-containing heterocyclic ring;

R ^{9 is defined} as the group R ³ but can not denote a hydrogen atom; or in which

R ¹ and R ⁸ together _ a C _{2, 0} or alkylidene mean by a hydroxy group substituted C2_, ₀ alkylidene:

A represents a cyclopentylene, cyclohexylene or a C ₂ _e alkylene group which is optionally substituted by one or more C 1-4 alkyl groups; R ^{2 represents} a hydrogen atom, an anionic charge or a common, readily removable carboxy protecting group, provided that when R ^{2 represents} a hydrogen atom or a protecting group, a counter anion is also present and R ' ^{4 is} a quaternized, nitrogen-containing one , aromatic or non-aromatic heterocycle bonded to A through a ring nitrogen atom to form a quaternary ammonium group, or a pharmaceutically acceptable salt thereof, to form an intermediate of general formula IV

COOR

IV

wherein R ¹ and R ^{8 have} the meanings given above, R ² 'is a conventional, easily removable carboxy-protecting group and L represents a common leaving group, with a thiol compound of general formula VII

HS-AR ¹⁴

Χ ^θ

VLI

wherein A and R ' ^{4 have} the meanings given above and X ^{e represents} a counter anion, in an inert solvent and in the presence of a base to a carbapenem compound.der the general formula Γ

S-A-R

\ 2 ¹

^X C00R

wherein R ¹ , R ⁸ , R ² ', A, R ¹⁴ and Χ ^{θ have} the meanings given above, and if desired, the carboxy-protecting group R ² ' removed to the corresponding deblocked compound of general formula I or a pharmaceutically acceptable To get salt from it.

The invention also relates to intermediate compounds of the general formula VU and to processes for preparing these intermediates.

The carbapenem compounds of general formula I are effective antibacterial agents or intermediates useful for preparing such agents.

The compounds of general formula I contain the carbapenem nucleus

and thus may be referred to as 1-carba-p-phen-S-carboxylic acid derivatives. However, the compounds may alternatively be considered to have the following basic structure

and are referred to as 7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylic acid derivatives. According to the invention compounds are included, wherein the hydrogen atoms in the 5- and 6-position can be present in both the cis and in the trans position, the preferred compounds having 5R, 6S (trans) configuration, as is the case with thienamycin , The compounds of the general formula I may be unsubstituted in the 6-position or substituted by the substituent groups described above for the other carbapenem derivatives. In particular, R ^{8 represents} a hydrogen atom and R ¹ may represent a hydrogen atom or a substituent other than hydrogen and disclosed, for example, in European Patent Application 38,869 (see the definition of R ₆ ). Alternatively, R ⁸ and R ¹ may together form a C ₂ _, o-alkylidene radical or a substituted (eg., By hydroxy) C2_ mean ₀ alkylidene. ^

The definitions for R ¹ and R ⁸ are described in more detail below:

(a) Aliphatic alkyl, alkenyl and alkynyl groups are straight-chain or branched groups having 1 to 10 carbon atoms, preferably 1 to 6 and especially preferably 1 to 4 carbon atoms. If these groups are part of another substituent, for example in a cycloalkyl radical or a heteroaralkyl or aralkenyl radical, then the alkyl, alkenyl and alkynyl group preferably contains 1 to 6 and particularly preferably 1 to 4 carbon atoms.

(b) By "heteroaryl" is meant mono-, bi- and polycyclic aromatic heterocyclic groups containing 1 to 4 O, N or S atoms, preferred are 5- or 6-membered heterocyclic rings, such as Thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, tetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, pyrazolyl, etc.

(c) "Heterocyclyl" is understood as meaning mono-, bi- and polycyclic, saturated or unsaturated, non-aromatic, heterocyclic groups which contain 1 to 4 O, N or S atoms. Preference is given to 5- or 6-membered, heterocyclic rings such as morpholinyl, piperazinyl, piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrrolinyl, pyrrolidinyl, etc.

(d) "halogen" refers to chlorine, bromine, fluorine and iodine atoms and preferably chlorine or bromine atoms.

The term "common, readily removable, carboxyl-protecting group" refers to a known ester group which has been used to block a carboxyl group during the chemical reactions described below and which can, if desired, be removed by methods which do not destroy the remainder of the molecule Examples of such ester-forming protecting groups are benzhydryl, p-nitrobenzyl, 2-naphthylmethyl, allyl, benzyl, trichloroethyl, for example chemical or enzymatic hydrolysis, treatment under mild conditions with chemical reducing agents, irradiation with UV light and catalytic hydrogenation. SiIyI, ζ B. trimethylsilyl, phenacyl, p-methoxybenzyl, acetonyl, o-nitrobenzyl, 4-Pyridylmethyi and C _1-6 -AlkVl, for example methyl, ethyl or t-butyl.As protective groups are also contemplated which are hydrolyzed under physiological conditions, such as pivaloylox ymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethyl. Particularly advantageous carboxyl-protecting groups are the p-nitrobenzyl group, which can be easily removed by catalytic hydrogenolysis, and the allyl group, which can be removed by Pd (PO ₃ ) 4-catalyzed reaction.

The pharmaceutically acceptable salts include the non-toxic acid addition salts, e.g. Salts with mineral acids such as hydrochloric acid, hydrobromic acid, iodic acid, phosphoric acid, sulfuric acid, etc., and salts with organic acids such as malic, acetic, citric, succinic, benzoic, tartaric, fumaric , Almond, ascorbic, lactic, gluconic and malic acid. Compounds of general formula I in the form of the acid addition salts can be described as

R ² = H or a protecting group

wherein Χ ^{θ represents} the acid anion. The counter anion X ^s can be chosen so that pharmaceutically acceptable salts are obtained for therapeutic purposes. In the case of the intermediate compounds of the formula I, however, X® can also be a toxic anion. In such a case, the ion may subsequently be removed or replaced with a pharmaceutically acceptable anion to obtain the active end product for therapeutic use. If acidic or basic groups are present in the group R ¹ or on the quaternized R ¹ * radical, the invention also includes suitable base or acid salts of these functional groups, ζ. B. acid addition salts with a basic group and metal salts (ζ. Β. Sodium, potassium, calcium and aluminum), the ammonium salt and salts with non-toxic amines (ζ B. trialkylamines, procaine, dibenzylamine, 1-ephenamine, N-benzyl-ß -phenethylamine, Ν, Ν'-dibenzylethylenediamine, etc.) in an acidic group. Compounds of general formula I, wherein R ^{2 represents} a hydrogen atom, an anionic charge or a physiologically hydrolyzable ester group, and the pharmaceutically acceptable salts thereof are useful antibacterial agents. The other compounds of general formula I are valuable intermediates which are disclosed in U.S. Pat above-mentioned, biologically active compounds can be converted.

Preferred compounds according to the invention are those compounds of the general formula I in which R ^{8 is} a hydrogen atom and R ^{1 is} a hydrogen atom, CH ₃ CH ₂ -,

, , ^ _v OH. OH

CH, C or CH ₅ CH- stands. CH ₃ ^

In this subclass are the preferred compounds

OH

those in which R ^{1 is} CH ₃ CH-. Particularly preferred compounds have the absolute configuration 5R, 6S, 8R. Preference is furthermore given to those compounds of the general formula I in which R ¹ and R ^e together form an alkylidene radical of the general formula

HIGH ₂

C =

CH ₃

form.

The alkylene or cycloalkylene radical A in the compounds of the general formula I can be a cyclopentylene radical

a cyclohexylene radical

or a G ^ -alkylene radical which is optionally substituted by one or more C ^ -alkyl substituents. Preferred substituents A are cyclopentylene, cyclohexylene or alkylene of the formula

R ¹⁰ R ¹²

C-

R ¹¹ R ¹³

in which R ¹⁰ , R ", R ¹² and R ' ³ each, independently of one another, denote a hydrogen atom or a C 1-4 -alkyl group, a preferred embodiment of which comprises compounds of the general formula I in which (ΙβΓ SIIb-)

stituent A is -CH ₂ CH ₂ -, -CHCH ₂ -, - \ ~} or -CH ₂ CH-

CH ₃ y - CH ₃

stands.

In some compounds of general formula I, when substituent A is a cycloalkylene or branched alkylene radical, one or more additional asymmetric carbon atoms are present resulting in the formation of diastereoisomers.

According to the invention, the mixtures of these diastereoisomers and also the individual, purified diastereoisomers

claimed.

The quaternized substituent R ^u is an optionally substituted mono-, bi- or polycyclic, aromatic or non-aromatic, heterocyclic radical containing at least one nitrogen atom in the ring and via a ring nitrogen atom on A

is bound, whereby a quaternary ammonium group is formed.

The formula stands for a substituted or unsubstituted, mono-, bi- or polycyclic heteroaryl radical which contains at least one nitrogen atom in the ring and is bonded via a ring nitrogen atom to a carbon atom of the substituent A, whereby

a quaternary ammonium group results. The heteroaryl group may be optionally substituted by the following substituents; C ^ -alkyl; C 1-4 alkyl substituted by hydroxy, amino, carboxy or halogen; C ₃ _s-cycloalkyl; Ci_4 alkoxy; C ^ alkylthio; amino; C ^ alkylamino; Di- (C, _4-alkyl) amino; Halogen; C, _4 alkanoylamino; C ^ alkanoyloxy; carboxy;

hydroxy; amidino; guanidino; trifluoromethyl; phenyl; Phenyl substituted by one, two or three amino,

-C-OC ₁ _ ₄ -alkyl}

Halogen, hydroxy, trifluoromethyl, C 1-4 alkyl or C 1-4 alkoxy groups; Heteroaryl and heteroaralkyl, wherein the heteroatom or heteroatoms in the abovementioned heterocyclic units are selected from 1 to 4 0, N or S atoms and the alkyl group connected to the heteroaralkyl group having 1 to 6 carbon atoms The heteroaryl group bonded to the substituent A. R ist is preferably a 5- or 6-membered aromatic heterocyclic radical containing a quaternized nitrogen atom bonded directly to a carbon atom of the alkylene or cycloalkylene moiety, and optionally one or more additional heteroatoms selected from O, N or S. Although generally each heteroaryl radical attached to A via a quaternized nitrogen atom results in biologically active carbapenem derivatives, however, a preferred embodiment of the invention comprises compounds wherein the radical of the general formula

- ¹ o

one of the radicals listed below represents:

: '' _R 7

wherein R ^B , R ⁶ and R ^{7 are} independently hydrogen; C ^ alkyl; C ₁ ^, - alkyl, substituted by hydroxy, amino, carboxy or halogen; Cj ^ cycloalkyl; C ₁ ^ -alkoxy; C ₁ _ _l alkylthio; amino; C ^ alkylamino; Di (C ₁ _ ₄ alkyl) amino; Halogen; C ₁ _4 alkanoylamino; Ci ^ i alkanoyloxy; carboxy;

: ό. -. .- ..: 'ν. , ..

_ _c alkyl;

hydroxy; amidino; guanidino; trifluoromethyl; phenyl; Phenyl substituted with one, two or three amino, halogen, hydroxy, trifluoromethyl, C _iJr alkyl or C _1jt alkoxy groups; and heteroaryl and heteroaralkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety attached to said heteroaralkyl moiety has 1 to 6 carbon atoms; or wherein two of R ⁵ , R ⁶ or R ⁷ together form a fused, saturated carbocyclic ring, a fused aromatic carbocyclic ring, a fused, saturated heterocyclic ring or a fused heteroaromatic ring;

or

which radicals are optionally substituted on a carbon atom by one or more substituents independently selected from the following radicals: C _1-4 -alkyl; C 1-4 alkyl substituted by hydroxy, amino, carboxy or halogen; Q j ^ -Cycloalkyi; C ₁ ^ -alkoxy; C ^ alkylthio; amino; C ^ alkylamino; Di (C _1-8 alkyl) amino; Halogen; C _1-4 -AlkBnOyIoXy; C ^ alkanoylamino; carboxy; O

-C-OC ₁ ^ -alkyl;

hydroxy; amidino; guanidino; trifluoromethyl; phenyl; Phenyl substituted by one, two or three amino, halo, hydroxy, trifluoromethyl, C ^ alkyl or C ₁ _ ₄ alkoxy; and heteroaryl or heteroaralkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety linked to said heteroaralkyl moiety has from 1 to 6 carbon atoms or, if desired, substituted such that a condensed carbocyclic or heterocyclic ring is formed;

(C)

or

these radicals being optionally substituted on a carbon atom by one or more substituents independently selected from the following radicals; C, _4 alkyl; C 1-4 alkyl substituted by hydroxy, amino, carboxy or halogen; C ^ cycloalkyl; C ₁ ^ -alkoxy; Ci_4-alkylthio; amino; Ci _ »- alkylamino; Di- (Ci_4-alkyl) amino; Halogen; C ^ -Alkahoylamino; Ci_4 alkanoyloxy; carboxy;

-C-OC ₁ _ ₄ -alkyl;

Hydroxy, amidino; guanidino; trifluoromethyl; phenyl; Phenyl substituted with one, two or three amino, halogen, hydroxy, trifluoromethyl, C 1-4 alkyl or C 1-12 alkoxy groups; and heteroaryl or heteroaralkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety attached to said heteroaralkyl moiety has 1 to 6 carbon atoms or are optionally so substituted, that a condensed carbocyclic or heterocyclic ring is formed;

(D)

or

which radicals are optionally substituted on a carbon atom by one or more substituents independently selected from the following radicals: C 1-4 alkyl; C 1-4 alkyl substituted by hydroxy, amino, carboxy or halogen; Q j ^ -Cycloalkyi; C, _4 alkoxy; C ₁ -alkylthio; amino; C, _4-alkylamino; Di (C _1-8 alkyl) amino, halogen, C, _M alkanoylamino; C, 4-alkanoyloxy, carboxy;

-C-Oc _1-4 __

hydroxy; amidino; guanidino; trifluoromethyl; phenyl; Phenyl substituted with one, two or three amino- ¹ , halogen, hydroxy, trifluoromethyl, C 1-4 -alkyl or C 1-4 -alkoxy groups; and heteroaryl or heteroaralkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety attached to said heteroaralkyl moiety has from 1 to 6 carbon atoms or are optionally substituted such that a condensed carbocyclic or heterocyclic ring is formed;

(e

or

worm Λ IUf Kj ₁ ouutuf mn äicui, wuuci η v> _1-4 - / - Mi \ yi uuoi riiciiyi ucucuko., wuuci uicoc ncaic vjcwuiiaunicmaiio at diicin carbon atom are substituted by one or more substituents independently of one another under the following radicals are selected: C ^ alkyl; C _1-4 alkyl substituted by hydroxy, amino, carboxy or halogen; C ^ cycloalkyl; C _1-4 alkoxy; C ^ alkylthio; amino; C _1-4 -alkyl! Amino; Di (C _1-4 -aikyl) amino; Halogen; C ^ alkanoylamino; C _1-4 alkanoyloxy; carboxy;

o -; .. "; '"

-C-OC _1-4 -alkyl;

hydroxy; amidino; guanidino; trifluoromethyl; phenyl; Phenyl substituted by one, two or three amino, halo, hydroxy, trifluoromethyl, Ci ^ alkyl or C ₁ _ ₄ alkoxy; and heteroaryl or heteroaralkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety linked to said heteroaralkyl moiety has from 1 to 6 carbon atoms or, if desired, substituted such that forming a fused carbocyclic or heterocyclic ring;

wherein X is O, S or NR, wherein R is C, _j-alkyl or phenyl, these radicals being optionally substituted on a carbon atom by one or more substituents which are independently selected from the following radicals: C _1-4 -alkyl; C 1-4 alkyl substituted by hydroxy, amino, carboxy or halogen; C ₃ _ä-cycloalkyl; C _1-4 alkoxy; C ^ alkylthio; amino; C ^ alkylamino; Di- (C ₁ ^ _r alkyl) amino; Halogen; C ₁ _ ₄ alkanoylamino; C ^ alkanoyloxy; carboxy;

-C-OC ₁ _ ₄ -alkyl;

hydroxy; amidino; guanidino; trifluoromethyl; phenyl; Phenyl substituted with one, two or three amino, halogen, hydroxy, trifluoromethyl, C 1-4 alkyl or C 1-4 alkoxy groups; and heteroaryl or heteroaralkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety linked to said heteroaralkyl moiety has 1 to 6 carbon atoms; and

N N-R

N N-R

1 Λ

wherein R is C 1-4 alkyl or phenyl, these radicals being optionally substituted on a carbon atom by one or more substituents independently selected from the following radicals: C 1-6 alkyl; Ci _ »- alkyl, substituted by hydroxy, amino, carboxy or halogen; C ^ -Cycloatkyl; C ₁ ^ ₁ -alkoxy; C - ^ - alkylthio; amino; C ^ alkylamino; Di (C _1-1 alkyl) amino; Ci_4 alkanoylamino; carboxy;

₁ _ ^

hydroxy; amidino; guanidino; trifluoromethyl; phenyl; Phenyl substituted with one, two or three amino, halogen, hydroxy, trifluoromethyl, C _1-4 alkyl or C _1-4 alkoxy groups; and heteroaryl or heteroaralkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety attached to said heteroaralkyl moiety has 1 to 6 carbon atoms. In the subclass above, the preferred compounds are those wherein the substituent A is -CH ₂ CH ₂ -, -CHCH ₂ -,

.CH ₃

- / \ or -CH ₂ CH-, and wherein - ^ CH,

* 1 p

(a) R and R together for HIGH ₂

stand or

(b) R ^{8 is} a hydrogen atom and R ^{1 is} a hydrogen atom, CH ₃ CH ₂ -,

CH ₃ OH OH

^, V ^ " ^{or GH} 3 ^CH " CH ₃ CH ₃

OH Particular preference is given to compounds in which R ^{8 is} a hydrogen atom and R ^{1 is} _CH 1 _H ^2.

wherein the preferred compounds have the absolute configuration 5R, 6S, 8R. According to a particularly preferred embodiment of the invention, compounds are obtained in which the radical of the formula

a remainder of the formula

in which R ^5, R ^e and R ^{7 are} independently a hydrogen atom, a C ^ alkyl group, a C ^ alkoxy group, a C ^ alkyl group substituted by a hydroxy group, a C ₁ ^ -AIkYlIhIOgTUpPe, amino , Carboxy and carbamoyl groups. In this subclass, the preferred compounds are those in which the substituent A is -CH ₂ CH ₂ -, -CHCH -

CH or -CH ₀ CH- and wherein either 3

CH, (a) R and R together for

HIGH ₂

C =

stand or '

(b) R ^{8 is} a hydrogen atom and R 'is hydrogen, CH ₃ CH ₂ -,

CH ₃ OH OH

^ - or CH ₃ CH- stands.

.r ° - OH

    *

Particular preference is given to compounds in which R ^{8 is} a hydrogen atom and R ^{1 is} CH ³ CH ³

stands. The preferred compounds have the absolute configuration 5R, 6S, 8R.

A most preferred embodiment of the invention involves the preparation of compounds wherein the radical of the general formula

- O

a radical of the general formula

R ⁶

R ⁷

in which R ⁵ , R ^e and R ⁷ independently of one another represent a hydrogen atom, a C _1-4 -alkyl, C _1-4 -alkoxy, C _1-4 -alkyl, C _1-4 -alkylthio and amino group which is substituted by a hydroxyl group. In this subclass, the preferred compounds are those in which the substituent A is -CH ₂ CH ₂ -,... CWPW.

CH ₃

or -CHpCH- and in which either CH,

(a) R ¹ and R ⁸ together for,

hocho

CH ₃

stand or

(b) R ^{8 is} a hydrogen atom and R ^{1 is} hydrogen, CH ₃ CH ₂ -,

CH ₃ CH ₃ OH OH

, CH-, ^ c- or CH ₃ CH- stands. CH ₃ CH ₃

OH Particular preference is given to compounds in which R ^{8 is} hydrogen and R 'is I

:. -CH ₃ CH-

stands. The preferred compounds have the absolute configuration 5R, 6S, 8R.

Another preferred embodiment of the invention involves the preparation of compounds wherein the radical of the general formula

\ + ^ S _{> T} SCR

a radical of the general formula N ~~ "\ ^" ^ - "

means; N ^. N-CH ₃

In this subclass, preferred compounds are those in which the substituent A is -CHCHp-, -CH ₂ CH ₂ -,

- / \ or -CH ₂ CH- CH,

\ / CH ₃ - *.

(a) R 'and R ⁸ to _S ammen for or -CH ₂ CH- Stehfe-WritftfOT-in either

HIGH ₂ - 3 - - ".-

CH,

stand or

(b) R ^{8 represents} a hydrogen atom and R ^{1 represents}

CH ₃ ' ^α 3 ^0H

Hydrogen, CH ₃ CH ₂ -, J ^c "- 2> ^c " ^or

CH ₃ CH ₃

OH CH ₃ CH- stands.

Especially preferred are compounds wherein R ^{8 is} a hydrogen atom and R ^{1 is} OH

CH ₃ CH- stands. The preferred compounds have the absolute configuration 5R, 6S, 8R.

A further preferred embodiment according to the invention comprises the preparation of compounds in which the radical of the general formula φ - ^ - ^.

-O

represents a pyridinium radical. In this subclass preferred compounds are those in which the substituent

tuent A is -CHCH ₂ -, -CH ₂ CH ₂ -, - \ _) or -CH ₂ CH- is CH_, ""

and in which either

(a) R 'and R ⁸ together for

HIGH _n

stand or

(b) R ^{8 represents} a hydrogen atom and R 'represents

CH ΓΗ ^0H

CH ₃ CH ₃ ^^ j

Hydrogen, CH ₃ CH ₂ ", CH-, J ^, G- or

CH, CH,

OH ^D

CH ₃ CH- stands.

Particularly preferred are compounds wherein R ^{8 is} a hydrogen atom and R ^{1 is} OH

CH ₃ CH- stands. The preferred compounds have the absolute configuration 5R, 6S, 8R.

A most preferred embodiment of the invention involves the preparation of compounds of the general formula

OK H

(R)

GOOR ⁴

Φ wherein the radical of the general formula -A-

has the following meanings:

(3) -CH ₂ CH ₂ -N

(S) -CH ₂ CH ₂ -N

(9) -CH ₂ CH ₂

(11) -CHCH ι CH-,

(2)

(4)

(10)

(12)

R- or S-diastereoisomers

^[ 2

CH

-CH-CH--

-CH ₂ CH ₂ -

CH ₂ OH

(6)

(8) -CH

2

CH.

-CH ₂ CH ₂ -

CH.

% = ^ ^SCn i

(13) -

or (14)

R, R or S / S diastereoisomers on the two asymmetric carbon atoms of the cyclohexyl group

and wherein R ^{2 represents} a hydrogen atom, an anionic charge or a common, easily removable carboxyl-protecting group, provided that, if R ^{2 represents} a hydrogen atom or a protecting group, there is also a counter anion, and the pharmaceutically acceptable acid addition salts thereof

A further preferred class of the quaternized R ^M substituents are those of the general formula

-N.

wherein R ^{16 is} selected from the following substituted and unsubstituted radicals: alkyl, alkenyl and alkynyl of 1 to 10 carbon atoms; Cycloalkyl and cycloalkylalkyl having 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moieties; phenyl; Aralkyl, araikenyl and aralkinyl, wherein the aryl moiety is phenyl and the aliphatic moiety has from 1 to 6 carbon atoms; Heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moieties attached to said heterocyclic moieties have 1 to 6 carbon atoms; wherein the abovementioned radicals R ^{16 are} optionally substituted by 1 to 3 substituents which are independently selected from the following radicals: C 1 -C 6 -alkyl, if desired substituted by amino, fluorine, chlorine, carboxyl, hydroxy or carbamoyl;

Fluorine, chlorine or bromine;

-OR ³ ; -OCO ₂ R ³ ; -OCOR ³ ; -OCONR ³ R ⁴ ; -OSO ₂ R ³ ;

oxo; -NR ^ R; R ^ CONR-; -NR ^ CO ^;

-NR ³ CONR ³ R ⁴ ; -NR ³ SO ₂ R ⁴ ; -SR ³ ; -SR ⁹ ; 0 0 -SR ⁹ ; -SO ₃ R ³ ; -CO ₂ R ³ ; -CONR ³ R ⁴ ; -CN; or

a phenyl radical which is optionally substituted by: 1-3-fluoro, chloro, bromo, d ^ -alkyl, -OR ³ , -NR ³ R ⁴ , -SO ₃ R ³ , -CO ₂ R ³ or -CONR ³ R ⁴ , wherein R ³ , R ⁴ and R ⁹ in these substituents R ^{16 have} the meanings given above; or wherein -v ---. R ^{16 is} a divalent phenylene or C, _4-Alkyiengruppe, which with the Ν ^φ J

Ring is connected so that forms a bridged, polycyclic group, and.

the rest of the general formula

a substituted or unsubstituted, mono-, bi- or polycyclic, non-aromatic, heterocyclic radical which may be fused to another aromatic or non-aromatic ring and which has at least one nitrogen atom in the ring and is bonded to A via a ring nitrogen atom, whereby a quaternary ammonium group. The heterocyclic radical may be saturated or unsaturated (having 1 to 2 double bonds) and may contain up to two further heteroatoms in addition to the quaternary nitrogen atom. Such additional heteroatoms are O, S (O) _n , N, NR ¹⁸ or NR ¹⁷ R ¹⁸ wherein m is 0.1 or 2, R ^{1S is} hydrogen, an optionally substituted C 1-4 alkyl or optionally substituted phenyl and R ¹⁷ and R ¹⁸ independently of one another are an optionally substituted Ci_s-alkyl or optionally substituted phenyl group. In a preferred embodiment, the radical is of the general formula

a non-aromatic, 4- to 7-membered, preferably 5- or 6-membered, N-containing heterocyclic ring having 0 to 2 double bonds and 0 to 2 additional heteroatoms, which is O, S (0) _m , N, NR ¹⁵ or NR ¹⁷ R ¹⁸ wherein m is 0.1 or 2, R ^{1S is} a hydrogen atom, a C 1-4 alkyl group which is optionally substituted by 1 to 2 substituents, these substituents being independently selected from -OR ³ , -NR ³ R ⁴ , -CO ₂ R ³ , oxo, phenyl, fluoro, chloro, bromo, -SO ₃ R ³ and -CONR ³ R ⁴ , or phenyl optionally substituted by 1 to 3 substituents, these substituents independently are selected from C 1-4 alkyl, -OR ³ ,. -NR ³ R ⁴ , fluoro, chloro, bromo, -SO ₃ R ³ , -CO ₂ R ² or -CONR ³ R ⁴ , and R ¹⁷ and R ' ⁸ each independently represent a C 1-4 alkyl group, which is optionally is substituted by 1 to 2 substituents, these substituents being independently selected from -OR ³ , -NR ³ R ⁴ , -CO ₂ R ³ , oxo, phenyl, fluorine, chlorine, bromine, -SO ₃ R ³ and -CONR ³ R ⁴ , or a phenyl group which is optionally substituted by 1 to 3 substituents, wherein these substituents are independently selected from C ^ alkyl, -OR ³ , -NR ³ R ⁴ , fluorine, chlorine, bromine, -SO ₃ R ³ , -CO ₂ R ² and -CONR ³ R ⁴ , wherein R ³ and R ⁴ in these heterocyclic NR ¹⁵ - or NR ' ⁷ R ¹⁸ groups have the meanings given above in connection with the substituent R' ⁶ . In such a preferred embodiment, the ring

desiredfails be substituted by 1 to 3 of the following substituents:

(a) C 1-8 alkyl, optionally substituted with 1 to 2 substituents independently selected from fluoro, chloro, bromo, -OR ³ , -OCOR ³ , -OCONR ³ R ⁴ , oxo, -NR ³ R ⁴ , -NR ³ COR ⁴ , -NR ³ CONR ³ R ⁴ , -NR ³ SO ₂ R ⁴ , -SR ³ , -SO ₃ R ³ , -CO ₂ R ³ and -CONR ³ R ⁴ ;

(b) _C2 ^ alkenyl, optionally substituted by 1 to 2 substituents independently selected from

Fluorine, chlorine, bromine, -OR ³ , -OCOR ³ , -OCONR ³ R *, oxo, -NR ³ R ⁴ , -NR ³ COR ⁴ , -NR ³ CONR ³ R ⁴ , -NR ³ SO ₂ R ⁴ , -SR *, -SO ₃ R ⁰ , -UU ₂ M "una -CONR ³ R ⁴ ;

(c) _C2 _8 alkynyl, optionally substituted by 1 to 2 substituents independently selected from fluoro, chloro / bromo, -OR ^3, -OCOR ^3, -OCONR ³ R ^4, oxo, -NR ³ R ⁴ , -NR ³ COR ⁴ , -NR ³ CONR ³ R ⁴ , -NR ³ SO ₂ R ⁴ , -SR ³ , -SO ₃ R ³ , -CO ₂ R ³ and -CONR ³ R ⁴ ;

(d) C _3-6 cycloalkyl, optionally substituted with 1 to 2 substituents independently selected from fluoro, chloro, bromo, -OR ³ , -OCOR ³ , -OCONR ³ R ⁴ , oxo, -NR ³ R ⁴ , ^ NR ³ COR ⁴ , -NR ³ CONR ³ R *, -NR ³ SO ₂ R ⁴ , -SR ³ , -SO ₃ R ³ , -CO ₂ R ³ and -CONR ³ R ⁴ ; , _.-. - _

(e) Cycioalkylalkyl having 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moiety, optionally substituted with 1 to 2 substituents independently selected from fluoro, chloro, bromo, -OR ³ , -OCOR ³ , -OCONR ³ R ⁴ , oxo, -NR ³ R ⁴ , -NR ³ COR ⁴ , -NR ³ CONR ³ R ⁴ , -NR ³ SO ₂ R ⁴ , -SR ³ , -SO ₃ R ³ , -CO ₂ R ³ and CONR ³ R ⁴ ;

(f) heteroaryl, wherein the heteroatom or heteroatoms are selected from 1 to 4 oxygen, nitrogen or sulfur atoms, optionally substituted by 1 to 2 substituents independently selected from fluoro, chloro, bromo, -OR ³ , - OCOR ^3, -OCONR ³ R ^4, oxo, -NR ³ R 4, -NR ³ COR ^4, -NR ³ CONR ³ R ^4, -NR ³ SO ₂ R ^4, -SR ^3, -SO ₃ R ^3, -CO ₂ R ³ and -CONR ³ R ⁴ ; preferred heteroaryl radicals are 5- or 6-membered, aromatic, heterocyclic rings;

(g) heteroaralkyl wherein the heteroatom or heteroatoms are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety has from 1 to 6 carbon atoms optionally substituted with 1 to 2 substituents independently selected from fluoro, chloro , Bromine, -OR ³ , -OCOR ³ , -OCONR ³ R ⁴ , oxo, -NR ³ R ⁴ , -NR ³ COR ⁴ , -NR ³ CONR ³ R ⁴ , -NR ³ SO ₂ R ⁴ , -SR ³ , -SO ₃ R ³ , -CO ₂ R ³ and -CONR ³ R ⁴ ; preferred heteroaralkyl groups are those wherein the heteroaryl group is a 5- or 6-membered aromatic heterocyclic ring and the alkyl moiety has 1 to 2 carbon atoms; ".".

(h) heterocyclyl, wherein the heteroatom or heteroatoms are selected from 1 to 4 oxygen, nitrogen or sulfur atoms optionally substituted by 1 to 2 substituents independently selected from fluoro, chloro, bromo, -OR ³ , - OCOR ³ , -OCONR ³ R ⁴ , oxo, -NR ³ R ⁴ , -NR ³ COR ⁴ , -NR ³ CONR ³ R ⁴ , -NR ³ SO ₂ R ⁴ , -SR ³ , -SO ₃ R ³ , CO ₂ R ³ and -CONR ³ R ⁴ ; preferred heterocyclyl groups represent 5- or 6-membered, saturated or unsaturated rings;

(i) heterocyclylalkyl wherein the heteroatom or heteroatoms are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety has 1 to 6 carbon atoms, optionally substituted with 1 to 2 substituents independently selected from fluoro, chloro , Bromine, -OR ³ Z-OCOR ³ , -OCONR ³ R ⁴ , oxo, -NR ³ R ⁴ , -NR ³ COR ⁴ , -NR ³ CONR ³ R ⁴ , -NR ³ SO ₂ R ⁴ , -SR ³ , -SO ₃ R ³ , -CO ₂ R ³ and -CONR ³ R ⁴ ; preferred heterocyclylalkyl radicals are those in which the heterocyclyl moiety is a 5- or 6-membered saturated or unsaturated ring, (j) fluoro, chloro or bromo;

(m) -OCOR ³ ; ,

(n) -OCONR ³ R ⁴ ; '

(o) -OSO ₂ R ³ ; - -

(p) oxo; (q) -NR ³ R ⁴ ;

(r) R ³ CONR ^4; ·

(s) -NR ³ CO ₂ R ⁴ ; (t) -NR ³ CONR ³ R ⁴ ; (u) -NR ³ SO ₂ R ⁴ ;

(v) -SR ³ ; -

O · ...

τ -. .

OO (x) -SR ⁹ ; ',

(Y) -SO ₃ R ³ ; , , :

(z) -CO ₂ R ³ ; (aa) -CONR ³ R ⁴ ; (bb) -CN; or

(cc) phenyl, optionally substituted by 1-3 fluoro, chloro, bromo, C, _s-alkyl - OR ³ Z-NR ³ R ⁴ , -SO ₃ R ³ , -CO ₂ R ³ or -CONR ³ R ⁴ , ,

The abovementioned substituents R ³ , R ⁴ and R ⁹ have the meanings given in connection with the substituent R ¹

 The above-defined ring.

is a non-aromatic, heterocyclic group. However, this ring may be fused to another ring containing a saturated or unsaturated carbocyclic ring, preferably a C 1-6 carbocyclic ring, a phenyl ring, a 4- to 7-membered, heterocyclic (saturated or unsaturated) ring containing 1 to 3 heteroatoms, which are selected from O, N, S (O) _m , NR ¹⁵ and NR ¹⁷ R ¹⁸ , or a 5- to 6-membered heteroaromatic ring containing 1 to 3 heteroatoms selected from O, S (0) _m , N, NR ' ⁵ and NR ¹⁷ R ^{18 are} selected, wherein m, R ¹⁵ , R ¹⁷ and R ^{18 have} the meanings given above. The substituent ^{16 of} the non-aromatic radical R ¹⁴ may be either (a) an optionally substituted C 1-4 alkyl, C 1-4

nyl, Cj_, _0- alkynyl, Cj ^ -cycloalkyl, (^ j-cycloalkyl-C ^ -alkyl, phenyl, phenyl-Ci_e-alkyl, phenyl-C ^ alkenyl, phenyl-C ^ alkynyl, heteroaryl, heteroaralkyl, wherein the alkyl moiety has 1 to 6 carbon atoms, heterocyclyl or heterocyclylalkyl group, wherein the alkyl moiety has 1 to 6 carbon atoms, or (b) represents a divalent phenylene group or C 1-4 alkylene group , which to the ring

fr

is bound to form a ring-bridged, polycyclic group, z. A quinuclidine group. The heteroaryl substituent (or the heteroaryl portion of the heteroaralkyl substituent) may be a mono-, bi- or polycyclic aromatic heterocyclic group containing 1 to 4 O, N or S atoms; preferred are 5- or 6-membered heterocyclic rings such as thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, tetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl and pyrazolyl. The heterocyclyl substituent (or the heterocyclyl portion of the heterocyclylalkyl substituent) may be a mono-, bi- or polycyclic, saturated or unsaturated, non-aromatic, heterocyclic group containing 1 to 4 O, N or S atoms; preferred are 5- or 6-membered heterocyclic rings such as morpholinyl, piperazinyl, piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrrolinyl and pyrrolidinyl.

If the substituent R ' ⁶ represents an alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, phenyl, phenylalkyl, phenylalkenyl, phenylalkynyl, heteroaryl, heteroaralkyl, heterocyclyl or heterocyclylalkyl group then these may If desired, groups may be substituted by 1 to 3 substituents which are selected from the following radicals:

(a) C _1-8 alkyl optionally substituted with preferably 1 to 3 amino, fluoro, chloro, carboxyl, hydroxy or carbamoyl groups;

(b) fluorine, chlorine or bromine;

(f) -OCONR ³ R ⁴ ;

(g) -OSO ₂ R ³ ; (h) oxo;

(i) -NR ³ R ⁴ ; (j) R ³ CONR ^4; (k) -NR ³ CO ₂ R ⁴ ; (I) -NR ³ CONR ³ R ⁴ ; (m) -NR ³ SO ₂ R ⁴ ; (n) -SR ³ ; (o) -SOR ⁹ ; (p) -SO ₂ R ⁹ ; , (q) -SO ₃ R ³ ; (r) -CO ₂ R ³ ; (s) -CONR ³ R ⁴ ; (t) -CN; or

(u) phenyl, optionally substituted by 1 to 3 substituents independently selected from fluoro, chloro, bromo, C 1-4 alkyl, -OR ³ , -NR ³ R ⁴ , -SO ₃ R ³ , -CO ₂ R ³ or CONR ³ R ⁴ , wherein in the above substituents R ' ^6, the groups R ³ and R ^{4 are} independently selected from hydrogen; Alkyl, alkenyl and alkynyl of 1 to 10 carbon atoms; Cycloalkyl, cycloalkylalkyl and alkylcycloalkyl having 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moieties; phenyl; Aralkyl, aralkenyl and aralkinyl, wherein the aryl moiety is phenyl and the aliphatic moiety has from 1 to 6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, wherein the heteroaryl and heterocyclyl group or part of this group is as previously defined for R ¹⁶ and the alkyl moieties associated with said heterocyclic moieties have from 1 to 6 carbon atoms; or R ³ and R ⁴ together with the nitrogen atom to which at least one of these radicals is bonded can form a 5- or 6-membered nitrogen-containing heterocyclic ring (as defined above for R ⁶ ); and R ^{9 has} the meanings given above for R ³ , but can not stand for hydrogen. The most preferred substituent R ¹⁶ is a C 1 e alkyl group, especially a methyl group.

If R 'is a divalent phenylene or ^a C, _ represents _a alkylene group, this group to another atom of the ring of the formula

bound to form a bridged polycyclic ring, e.g. B. a quaternized quinuclidine ring of the formula

According to a particularly preferred embodiment, compounds of the general formula 1 are prepared according to the invention, wherein the radical of the formula '

has the following meanings:

/ -ν

CH.

N-CH.

CH.

S- »0

CH.

.CH.

CH.

or

wherein Y represents a hydrogen, chlorine, bromine, iodine or fluorine atom or a C ^ alkyl, hydroxy, -SC ^ alkyl, carboxyl, carbamoyl or phenyl group. In this subclass, preferred compounds are those,

wherein A is -CHCH ₂ -, -C ~ \ * -CH ₂ CH- or - (CH ₂ ) _n , CH ₃ ^ ~ ^ CH ₃

wherein η is 2, 3 or 4, and especially those

wherein A is -CH ₂ CH ₂ -, -CH ₂ CH ₂ CH ₂ -, -CHCH ₂ -, - {~~) or

-CH ₂ CH-, and most preferably those wherein A is CH,

is -CH ₂ CH ₂ - and wherein either 1S

(a) R and R together for

HIGH,

C =

CH,

stand or

(b) R ^{8 represents} a hydrogen atom and R 'represents

CH,

CH, OH

Hydrogen, CH ₅ CH ₃ , CH-,

OH C- or CH, CH-

CH-

CH-

stands.

Particularly preferred are compounds wherein R ^{8 is} a hydrogen atom and R 'is OH

CH3CH-, in particular, compounds of the absolute configuration 5R, 6S, 8R are preferred.

In a more preferred embodiment, compounds of the general formula I are prepared according to the invention, wherein the radical of the general formula

has the following meanings:

CH

N

N-CH,

"3 / -V,

or

In this preferred subclass, preferred compounds are those wherein A is

-CHCH ₂ -,

, -CH ₂ CH-

CH ₃ N / CH ₃

- (CH ₂ ) ,, -, in which η is 2, 3 or 4, more preferably those in which A is -CH ₂ CH ₂ -, -CH ₂ CH ₂ CH ₂ -,

-CHCHo-, ι CH ₃

or -CH ₂ CH-I CH ₃

and, most preferably, those wherein A is -CH ₂ CH ₂ -, and wherein either (a) R ¹ and R ⁸ together represent

HIGH,

CH,

C =

stand or

(b) R ^{8 represents} a hydrogen atom and R 'represents

Hydrogen,

CH ₃ CH ₃ OH ^ CH-,> C

CH

stands.

CH ₃

OH or CH ₃ CH-

Especially preferred are compounds wherein R ^{8 is} a hydrogen atom and R 'is

? ^H CH ₃ CH -, in particular compounds of the absolute configuration 5R, 6S, 8R.

In a further, more preferred embodiment, compounds of the general formula I are prepared according to the invention, wherein the radical of the formula ^:

_n i6

₃ -N

wherein Y is a hydrogen, chlorine, bromine, iodine or fluorine atom or a C ^ alkyl, hydroxy, -S-Ci ^ alkyl, carboxyl, carbamoyl or phenyl group. In this preferred subclass, the preferred compounds are those wherein A is - (CH ₂ J _n -, where η is 2, 3 or 4. Most preferred are those compounds wherein A is -CH ₂ CH ₂ - and in which either

(a) R ¹ and R ⁸ together for

HIGH ₂ CH ₃

stand or

(b) R ^{8 represents} a hydrogen atom and R ^{1 represents}

CH ₃ CH ₃ OH OH hydrogen, CH ₃ CH ₂ -, CH- ;, C or CH ₃ CH-

stands. /: ·. ' '

Particular preference is given to compounds in which R ^{8 is} a hydrogen atom and R '

CH ₃ CH-, in particular compounds of the absolute configuration 5R, 6S, 8R.

In a most preferred embodiment of the invention compounds of general formula I are obtained, wherein the radical of formula

stands.

In this preferred subclass, preferred compounds are those wherein A is - (CH ₂ J _n - wherein η is 2, 3 or 4, and most preferably those wherein A is -CH ₂ CH ₂ - and wherein either

(a) R ¹ and R ⁸ together for

HIGH ₀

C = CH ₃

stand or

(b) R ^{8 is} a hydrogen atom and R 'is for

CH, CH, OH OH

stands.

Hydrogen, CH ₃ CH ₂ -CH-, C- or CH ₃ CH-

CH _x ^ CH,

Especially preferred are compounds wherein R ^{8 is} a hydrogen atom and R 'is

CH3CH-, in particular compounds of the absolute configuration 5R, 6S, 8R.

In a most preferred embodiment, compounds of the general formula are obtained according to the invention

R ¹⁶

S-CH ₂ CH ₂ -N

COOR "

wherein

For

CK

N-CH, or

(both α- and β-diastereoisomers)

stands

and wherein R ^{2 represents} a hydrogen atom, an anionic charge, or a common, readily removable carboxyl-protecting group, provided that if R ^{2 represents} a hydrogen atom or a protecting group, then there is also a counterion, and the pharmaceutically acceptable addition salts thereof ,

Certain compounds covered by general formula I may exist both as optical isomers and as epimer mixtures. According to the invention, all of these optical isomers and the epimer mixtures are included. For example, if the 6-position substituent is a hydroxyethyl group, then this substituent may be in both the R and S configurations. Thus, according to the invention, both the obtained isomers and their epimer mixtures are included. In the process according to the invention, an intermediate compound of the general formula IV is used

CCOR

IV

a, which is disclosed for example in European Patent Application 38 869 and which can be prepared by the general method described therein. L is a common leaving group (it is referred to in the European patent application 38 with "X" ·), such as chlorine, bromine, iodine, benzenesulfonyloxy, p-toluenesulfonyloxy, p-nitrobenzenesulfonyloxy, methanesulfonyloxy, trifluoromethanesulfonyioxy, diphenexyphosphinyloxy or di- (trichloroethoxy) - The preferred leaving group is diphenoxyphosphinyloxy The intermediates of general formula IV are generally formed in situ by reaction of an intermediate compound of general formula U1

worm K ', It "Una rr aie ooen anyByeueiioi gcucuiunycn ucgiu.su, <». i wn> ». <> ₉ u _U , _g ..» .., _o _ _. ... _. _w

Intermediate IV wherein L is a diphenoxyphosphinyloxy group can be prepared by reacting a ketoester of Formula III in an inert organic solvent such as methylene chloride, acetonitrile or dimethylformamide with about one molar equivalent of diphenylchloride phosphate in the presence of a base such as diisopropylethylamine, triethylamine, 4 dimethylaminopyridine or the like., is reacted at a temperature of about -20 ° to + 4O ⁰ C, most preferably at etwaO ° C. The intermediate compound IV can be isolated if desired; However, it is usually used without further isolation or purification as a starting compound for the process according to the invention.

In the process according to the invention, a carbapenem intermediate IV is reacted with a quaternary amine thiol compound of the general formula VII

HS-A-R ' ⁴ Χ ^θ VII

wherein A is a cyclopentylene, cyclohexylene or (C 1-4) alkylene group which is optionally substituted by one or more C 1-4 -alkyl groups, most preferably a cyclopentylene group, a cyclohexylene group or

R ¹¹ R ^13;

wherein R ^{1 is} VR ", R ¹² and R ^{13 are} each independently of one another a hydrogen atom or a C 1-4 -alkyl group, X ^{8 is} a counterion which is derived from a strong acid, for example 'Cl', Br ',

V. .CH, SO, ", CF * SO," or .CH, - ^ .- \ S-so, ", and R one

D O J J J \ -J $

as defined above, quaternized, nitrogen-containing, aromatic or non-aromatic heterocycle. The reaction is carried out in an inert solvent, such as acetonitrile, acetonitrile-dimethylformamide, tetrahydrofuran, tetrahydrofuran-H ₂ 0, acetonitrile-H ₂ O or acetone, in the presence of a base. The type of base is not critical. However, the best results are obtained when using a non-nucleophilic tertiary amine as the base, such as diisopropylethylamine,!, S-diazabicycloWSAQJundec-T-ene, 1,5-diazabicyclo [4.3.0] -non-5-ene, or a tri (C 1-4) alkylamine, such as triethylamine, tributylamine or tripopylamine. The reaction of the intermediate IV with the thiol VII can be carried out in a wide temperature range, for. B. at -15 ⁰ C bisizu room temperature, perform. Preferably, one operates in a temperature range of about -15 ° to +15 ⁰ C, most preferably about 0 ° C.

The carbapenem compound obtained by reacting the quaternary aminethiol of the formula VII with the intermediate IV has a counter anion [e.g. (CeH ₅ O) ₂ PO ₂ ⁸ , Cl ^e or the anion associated with the quaternary thiol], the compound associated with the compound] associated with the compound. This counter anion can be replaced at this stage by conventional methods by another Gegenanion, z. One that is more pharmaceutically acceptable. Alternatively, one can remove the counter anion during the subsequent deblocking step: If the quaternized carbapenem compound and the counter anion form an insoluble product, then the product can be allowed to crystallize out as it is formed and can be recovered by filtration.

After obtaining the desired carbapenem compound, the carboxyl protecting group R ² 'of compound Γ can be removed, if desired, by conventional methods such as solvolysis, chemical reduction or hydrogenation. When the protecting group is a p-nitrobenzyl, benzyl, benzhydryl or 2-naphthylmethyl group which is readily removed by catalytic hydrogenation, the intermediate compound I 'is treated in a suitable solvent such as dioxane-water-ethanol , Tetrahydrofuran-diethyl ether buffer, tetrahydrofuran-aqueous dipotassium hydrogenphosphate isopropanol or the like, at a hydrogen pressure of 1 to 4 at in the presence of a hydrogenation catalyst such as palladium on carbon, palladium hydroxide, platinum oxide or the like, at a temperature of from 0 to 50 ⁰ C for about 0.24 to 4 hours. If the group R ² is o-nitrobenzyl, photolysis can be used for deblocking, protective groups such as 2,2,2-trichloroethyl can be removed by mild reduction with zinc. The allyl protecting group can be removed using a catalyst such as a mixture of a palladium compound and triphenylphosphine in a suitable aprotic solvent such as tetrahydrofuran, methylene chloride or diethyl ether. Similarly, other conventional carboxyl protecting groups can be removed by methods known to those skilled in the art. Finally, as stated above, the compounds of the formula Γ in which R ² 'is a physiologically hydrolyzable ester group, such as acetoxymethyl, phthalidyl, indanyl, pivaloyloxymethyl, methoxymethyl, etc., can be administered directly to the host without deblocking such esters are hydrolyzed in vivo under physiological conditions. If the substituent R ¹ and / or R ⁸ or the quaternized nucleophile R ¹⁴ bound to the substituent A contains a functional group which might interfere with the intended course of the reaction, then this group can be protected with a conventional blocking group and then subsequently deblocked to give the group to regenerate the desired functional group. Suitable blocking groups and methods for introducing and removing such groups are well known to those skilled in the art. · '

As in other β-lactam antibiotics, the compounds of the general formula I can be converted by known methods into pharmaceutically acceptable salts, which for the purposes according to the invention are essentially equivalent to the compounds not converted into the salts. Thus, for example, a compound of the general formula I in which R ^{2 is} an anionic charge can be dissolved in a suitable inert solvent. Then one adds one equivalent of a pharmaceutically acceptable acid. The desired acid addition salt can be prepared by conventional methods, for. As solvent precipitation, lyophilization, etc. win. Lying in the compound of general formula I other basic or

acidic, functional groups, one can obtain the pharmaceutically acceptable base addition salts and acid addition salts in a similar manner by known methods.

The compounds of the general formula I in which R ² denotes a hydrogen atom or an anionic charge, or pharmaceutically acceptable salts thereof, can also be converted by customary processes into the corresponding compounds in which R ^{J is} a physiologically hydrolyzable ester group. Compounds of the general formula I in which R ^{2 is} a conventional carboxyl-protecting group can be converted into the corresponding compounds in which R ^{2 is} a hydrogen atom, an anionic charge or physiologically hydrolyzable ester group or in the pharmaceutically acceptable salts thereof.

The carbapenem derivatives of the general formula I, wherein R ^{2 represents} a hydrogen atom, an anionic charge or a physiologically hydrolyzable carboxyl-protecting group, or the pharmaceutically acceptable salts thereof are effective antibiotics which can be used against various Gram-positive and Gram-negative bacteria. They can also be used, for example, as animal feed additives to increase growth, as preservatives for food !, as bactericides for industrial applications, eg. In water-based paints or in the wash water of paper mills for inhibiting the growth of dangerous bacteria, and as a disinfecting agent for destroying or inhibiting the growth of dangerous bacteria on medical and dental equipment. However, they are particularly useful for treating human or animal infectious diseases caused by Gram-negative and Gram-positive bacteria.

The pharmaceutically active compounds provided by the invention may be used alone or formulated as pharmaceutical agents. Such agents contain, in addition to the active carbapenem compound, a pharmaceutically acceptable carrier or diluent. The compounds can be administered in a variety of ways. These include in particular oral, topical or parenteral (intravenous or intramuscular injection) administration. The pharmaceutical agents may be in solid form, e.g. As capsules, tablets, powders, etc., or in liquid form as solutions, suspensions or emulsions. Agents for infections (preferred mode of administration) may be prepared in unit dosage form in ampoules or may be in multi-dose containers and may contain formulatory agents such as suspending agents, stabilizing agents or dispersing agents. The agents may be in ready to use form. They may also be in powder form so that at the time of administration they may be supplemented with a suitable carrier, e.g. As sterile water, reconstituted.

The dose to be administered will depend primarily on the compound used, the particular formulation, the mode of administration of the nature and the condition of the host. Of course, the dosage also depends on the specific situs and the organism to be treated. The choice of the most preferred dosage and the route of administration is left to the treating physician, but in general the compounds are administered parenterally or orally to a mammal (human and animal) in an amount of about 5 to 200 mg / kg / day. Administration is generally in several doses, ζ. B. three or four times a day

 The following examples serve to illustrate the invention.

example 1

Preparation of 3- (2- (1-pyridinium) ethylthio) -6a- {1- (R) -hydroxyethyl) -7-oxo-1-azabicyclo {3.2.0} hept-2-ene-2-carboxylate A. 1- (2-mercaptoethyl) pyridinium methanesulfonate

55 ⁰ C, 16 h

A suspension of pyridinium methanesulfonate in pyrene is prepared by dropwise adding methanesulfonic acid (1.95 ml, 0.03 mol) to pyridine (8.0 ml, 0.099 mol) with cooling. To this suspension is added ethylene sulfide (1.96 ml, 0.033 mol). The resulting mixture is stirred for 16 h at 55 ⁰ C, concentrated under reduced pressure to a thick syrup, which is mixed with less ml of water and the solution is poured onto a column (40 x 16 cm) of μ-Bondapak C-18, which is eluted with water. The lyophilization of

suitable fractions results in a colorless syrup; Yield 6.5 g (91%).

IR (FiIm) v _max : 2300-2600 (br, SH), 1 635 (pyridinium), 1490, 1200 (sulfonate), 1068, 1060, 1045, 791.780 cm ^{-1 1} H-NMR (DMSO-d _e ) δ: 2.32 (3H, s, CH ₃ SO ₃ -), 2.61, 2.70, 2.73, 2.82 (1H, B part of the A ₂ B system, SH), 3.07 (2H, m [with D ₂ O: 3.08 (2H, t, J = 6.5 HZ)], CH ₂ S), 4.76 (2H, t, J = 6.5 Hz, CH ₂ N-), 8.19 (2H, m, Hm of pyridinium), 8.6 (1H, m, Ho of pyridinium), 9, L8

(2H, dd, J = 6.8 Hz, J = 1.4 Hz, Ho of pyridinium) UV (H ₂ O) X _max : 206 (ε 5230), 258 (ε 3760) ηημ. Method A B. 1- (2-Mercaptoethyl) -pyridinium chloride ·

' _Λ - permutite S-1 Cl "-MsO --- ¥

An aqueous solution of crude 1- (2-mercaptoethyl) -pyridinium methanesulfonate (9.4 g, 0.04 mol) is added to a column (2.5 x 41 cm) of permutite S-1 Cl " eluting at a rate of 0.5 ml / min, combine the appropriate fractions and lyophilize to give a yellow syrup, yield 7.0 g (100%), which is thus used for the next step.

'H NMR (D ₂ O) δ: 3.22 (2H, m, CH ₂ S), 4.88 (m, CH ₂ N ⁺ ), 8.18 (2H, m, Hm of pyridinium), SJ (1H, m, Hp of pyridinium), 9.0 ppm (2H, m, Ho of pyridinium).

Method B

Cl

CI

To pre-cooled (ice bath) pyridine (5.6 mL, 70 mmol) is added pyridine hydrochloride (4.05 g, 35 mmol) and ethylene sulfide (2.1 mL, 35 mmol). The mixture is heated to 65 ⁰ C and stirred for 75 min, to obtain a two-phase system. The lighter phase is removed. The remaining oil is washed with ether (5 x 10 ml) and a high vacuum is applied to give the title compound (90-100%) which is then used for the next step.

C. p-Nitrobenzyl-3- [2- {1-pyridinium) ethylthio] -6a [1- {R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene 2-carboxylate chloride

• OH

1)

2) ClP (GPh) ₂

COGPNB 4) NEt (IPr) ₂

COOPNB

A solution of p-nitrobenzyl 6α- [1- (R) -hydroxyethyl] -3,7-dioxo-1-azabicyclo [3.2.0] heptane-2-carboxylate (6.09 g, 17.5 mmol) in acetonitrile (20 ml) is cooled under nitrogen atmosphere to +5 ⁰ C, treated sequentially with diisopropylethylamine (3.65 ml, 21, OmMoI) and diphenyl chlorophosphate (4.34 ml, 21.0 mmol), the mixture obtained is stirred for 30 min at 5 ° C, cooled to -5 ⁰ C and treated sequentially with a solution of crude 1- (2-mercaptoethyl) pyridinium chloride (4.3 g, 24 mmol) in Ν, Ν-dimethylformamide (1.0 ml) and dropwise with diisopropylethylamine (3.65 ml, .21.0 mmol). The reaction mixture is stirred for 1 h at ^{0 0} C, cooled to -30 ⁰ C and stirred for a further 15 min. The solid is filtered off and washed with cold (-3O ⁰ C) acetonitrile to give 5.77 g (65%) yield

IR (nujol) v _max : 3300 (OH), 1775 (C = O of β-lactam), 1690 (C = Odes PNB ester), 1630 (pyridinium), 1 605 (phenyl of the PNB ester),

1515 (NO ₂ ), 1335 Cm- '/ ·'

1 H-NMR (DMSO-d _e ) δ: 1.17 (3H, d, J = 6.1 Hz, CH ₃ CHOH), 3.2-3.75 (5H, H-4, H-6, CH ₂ S), 3.75-4.5 (2H, H-5, CH ₃ CHOH), 4.92 (2H,

br.t, J = 6.5 Hz, CH ₂ N ⁺ ), 5.18 (1H, d, J ~ 4.9 Hz, OH), 5.37 (center of ABq, J _a , "= 14, 2 Hz, CH ₂ , from PNB), 7.69 (2H, d, J = 8.7 Hz, Ho from PNB), 8.24 (d, J = 8.7 Hz, Hm from PNB), 8, 0-8.4 (4H, Hm of PNB, Hm of pyridinium), 8.66 (1H, m, Hp

of pyridinium), 9.17 (2H, br.d, J - 5.5 Hz, pyronium, Ho).

The filtrate and the solvent used for washing are combined and diluted with 150 ml of ether. The supernatant liquid is decanted and the gum product dissolved in 40 ml of water containing acetonitrile sufficient to give a solution which is applied to a column (3 × 10 cm) of μ-Bondapak C-18. The column is eluted with 10% acetonitrile-90% water (150 ml) and 50% acetonitrile-50% water (100 ml) mixtures. The appropriate fractions are combined and lyophilized after removing the acetonitrile in vacuo. A yellow powder is obtained. The NMR shows the presence of the title compound with something

p-nitrobenzyl-3- [2- (1-pyridinium) -ethylthiOJ-6a- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate Diphenyl phosphate (2: 1) is mixed. Dissolve the powder in as little water as possible and add water over a column (1.5 x 21 cm) of Permutit S-1 Cl ". Lyophilization of the appropriate fractions gives 1.8 g (20%) of the title compound ,

D. p-Nitrobenzyl-S - ^ - ii -pyridinium-1-ethylthio-1-yl-1-R, -Hydroxyethyl-T-oxo-i-azabicyclo [s] .Olhept ^ -en-1-carboxylate diphenyl phosphate

1) NEt (IPr) ₉

0 ^

2) ClP (OPh) ₂

OH

COOPNB

4) NEt (iPr).

COOPNB

A solution of p-nitrobenzyl-6a [1- (R) -hydroxyethyl] -3,7-dioxo-1-azybicyclo [3.2.0] heptane-2-carboxylate (0.174 g, 0.50 mmol) in acetonitrile ( 2 ml) is cooled under nitrogen atmosphere to 0 ° C, treated successively with diisopropylethylamine (0.105 ml, 0.60 mmol) and diphenylchlorophosphate (0.124 ml, 0.60 mmol); the resulting solution is stirred at 0 ^° C. for 30 minutes and treated successively with a solution of 1- (2-mercaptoethyl) pyridinium methanesulfonate (0.170 g, 0.72 mmol) in 0.6 ml acetonitrile and diisopropyl ethylamine (0.105 ml , 0.60 mmol). the reaction mixture is stirred for 15 min at 0 ° C. the mixture is diluted with cold (0 ⁰ C) water (7 ml) and the mixture is applied to a column (1.5 χ 6.4 mm) of μ-Bondapak C-18 The column is eluted with a mixture of acetonitrile (25% -50%) in water (75% -50%) The appropriate fractions are combined and lyophilized after removing the acetonitrile in vacuo. A yellow powder is obtained, 0.33 g (92%).

IR (KBr) vmax: 3600-3000 (OH), 1765 (C = O of β-lactam), 1690 (C = O of the PNB ester), 1625 (pyridinium), 1585 (phenyl), 1510 (NO ₂ ), 1330 (NO ₂ ), 885 cm "'(NO ₂ )

¹ H-NMR (DMSO-d ₉ ) δ: 1.16 (3H, d, J = 6.2 Hz, CH ₃ CHOH), 4.87 (2H, br.t, J = 6.6 Hz, CH ₂ S), 5.37 (center of ABq, J _a , _b = 14.3 Hz, CH ₂ of PNB), 6.7-7.5 (phenyl), 7.68 (d, J = 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, J = 6.7 Hz, J = 1.3 Hz, Ho of pyridinium).

E. 3- {2- (1-Pyridinium) ethylthio] -6a [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate Method A

OH

10% Pd / C, H ₂ THF, ether

COOPNB

COO

To a solution of

p-nitrobenzyl-3- [2- (1-pyridinium) -ethyithio] -6a- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo) o- [3.2.0] hept-2-ene Ethereal (10 ml), potassium phosphate monobasic sodium hydroxide buffer pH 7.4 (16 ml, 0.05.), Is added ethereally (10 ml) to wet tetrahydrofuran (10 ml) in dry tetrahydrofuran (10 ml) M) and 10% palladium on carbon (0.16 g). The resulting mixture is hydrogenated at 25 ^° C. and 40 psi for 1 h. Separate the two phases and wash the organic phase with water (2x3 ml). The aqueous solutions are combined, washed with ether (2 x 10 ml) and added to a column (1.5 x 6.2 cm) of μ-Bondapak C-18 after removing traces of the organic solvents in vacuo. The column is eluted with water and, after lyophilization of the appropriate fractions, a yellow powder is obtained, yield 0.062 g (84%).

IR (KBr) v _max : 3700-3000 (OH), 1755 (C = O of β-lactam), 1630 (pyridinium), 1590 cm ^-1 (carboxylate) 'H-NMR (D ₂ O) δ: 1, 22 (3H, d, J = 6.4 Hz, CH ₃ CHOH), 2.92 (d, J = 9.1 Hz, H-4), 2.97 (d, J = 9.1 Hz, H -4), 3.20 (dd, J = 2.5 Hz,

J = 6.1 Hz, H-6), 3.44 (t, J = 6.0 Hz, CH ₂ S), 3.93 (dd, J = 9.1 Hz, J = 2.5 Hz, H-5), 4.82 (t, J = 6.0 Hz ', CH ₂ N ⁺ ), 8.04 (m, Hm of pyridinium), 8.5 (m, Hp of pyridinium), 8.82 (dd, J = 3.2 Hz, J = 1.1 Hz, Ho of pyridinium) UV (H ₂ O) λ ™ ,: 259 (ε 5800), 296 (ε 7030) mM

τ _{/ 2} = 13.5 (measured at a concentration of 10 ^"4 M in phosphate buffer pH 7.4 at 36.8 ⁰ C) h.

Method B

OH

1096 Pd / C, H ₀

THF, ether, buffer 7.2

COOPNB

COO

To a solution of

p-Nitrobenzyl-3- [2- (1-pyridinium ") ethylthio] -6a [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene Add 2-carboxylate chloride (5.77 g, 11.4 mmol) in potassium phosphate monobasic sodium hydroxide buffer (170 mL, 0.2 M, pH 7.22) to tetrahydrofuran (30 mL), ether (30 mL). and 10% palladium on carbon (5.7 g), hydrogenate the resulting mixture for 1 h at 22 ° C and 2.76 bar (40 psi) and filter through a pad of Celite. Wash the pad with water (2x15 ml), combine the filtrate and the washings and dilute with ether (100 ml), separate the aqueous layer, wash with ether (3 × 100 ml) and pour onto a column (4.5 × 20 cm) of μ- Bondapak C-18, after removing the organic solvents in vacuo, eluting the column with water and then with a mixture of 1% acetonitrile in water to give, after lyophilization of the appropriate fractions, 2.48 g (65%) of the title compound as a yellow powder, the analytical data are identical to those for the compound prepared according to method A.

Example 2 Preparation of

COO

A. 1- {2-Mercaptoethyl) -3,5-dimethylpyridinium methanesulfonate

MsOH

MsO "

To a suspension of 3,5-lgtidinium methanesulfonate in 3,5-lutidine prepared by adding methanesulfonic acid (0.65 ml, 0.010 mol) to cold 3,5-lutidine (2.51 ml, 0.022 mol) ethylene sulfide (0.655 ml, 0.011 mol), the resulting mixture is stirred at 55 ° C under a nitrogen atmosphere for 24 h, cooled to 23 ° C and diluted with water (5 ml) and ether (5 ml). The organic layer is separated off and the aqueous solution is washed with ether (6 × 4 ml). The traces of ether are removed in vacuo and the solution is applied to a column (2.5 × 6.0 cm) of μ-Bondapak C-18. The column is eluted with water and lyophilizes the appropriate fractions to give a colorless syrup, yield 2.4 g (91%).

IR (film) v _mall : 2520 (SH), 1 628 (pyridinium), 1,600, 1495, 1 325, 1 305, 1 283, 1 200 (sulfonate), 1040, 938, 765, 680 cm ^-1 H-NMR (DMSO-d _e ) 5: 2.31 (3H, s, CH ₃ SO ₃ -); 2.47 (6H, s, CH ₃ of pyridinium), 2.57, 2.66, 2, 69, 2.78 (1Ή, B part of the A ₂ B system, SH), 3.06 (2H, m [with added D ₂ O (2H, t, J = 6.5 Hz)], CH ₂ S), 4.65 (2H, t, J = 6.5 Hz, CH ₂ N ⁺ ), 8.34 (1H, s, Hp of

Pyridinium), 8.79 (2H, s, Ho of pyridinium) UV (H ₂ O) λ ™ _χ : 271 (ε 4860) πημ Analysis: Calculated for C ₁₀ H ₁₇ NO ₃ S ₂ .0.5H ₂ O 44 , 09% H 6.66% N 5.14% S 23.54% found: 44.26 6.49 5.17 24.18,

For example, p-nitrobenzyl-3- [2- (1- (3,5-dimethylpyridine)) ethylthio] -6a- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0. ] hept-2-ene-2-carboxylate diphenyl phosphate. ,

1) NEt (iPr)

O π

COOPNB

2) ClP (OPh) ₂

4) NEt (iPr) ₂

COOPNB ₍ p _h0) J ₀ -

(1- (R) -hydroxyethyl) To a cold ⁽⁰ ° C) solution of p-nitrobenzyl 6a- -3,7-dioxo-1-azabicyclo [3.2.0] heptane-2-carboxylate (0.523 g, 1 , 50 mmol) in acetonitrile (6.0 ml) are added under nitrogen atmosphere diisopropylethylamine (0.314 ml, 1.8 mmol) and then diphenylchlorophosphate (0.373 ml, 1.8 mmol). The reaction mixture is stirred for 30 minutes and treated with a solution of 1- (2-mercaptoethyl) -3,5-dimethylpyridinium methanesulfonate (0.493 g, 1.87 mmol) in acetonitrile (1.9 ml) and then with diisopropylethylamine ( 0.314 ml, 1.8 mmol). The reaction mixture is stirred for 1 h at ⁰ ° C, diluted with cold (0 ⁰ C) water (26 ml) and poured into a column (7.0 x 3.5 cm) of μ-Bondapak C-18th The column is eluted with a 25-50% Aceonitrii 75-50% water mixture and, after lyophilization of the appropriate fractions, 1.01 g (90%) of the title compound is obtained as a yellow powder. IFt (KBr) v _max : 3700-3100 (OH), 1778 (C = O of β-lactam), 1700 (C = O of the PNB ester), 1 635 (pyridinium), 1 595 (phenyl), 1 521

(NO ₂ ), 1335 (NO ₂ ), 895 cm ^{-1 1} H-NMR (DMSO-d ₆ ) &: 1.16 (3H, d, J = 6.1 Hz, CH ₃ CHOVi), 2.43 (s, CH ₃ of pyridinium), 4.75 (2H, m, CH ₂ N -), 5.38 (center of ABq, J ₃ ,, = 14.3 Hz, CH ₂ of PNB), 6.6 -7.5 (10H, m, phenyl), 7.70 (2H, d, J = 8.7Hz, Ho of PNB), 8.0-8.5 (3H, m, Hp of

Pyridinium, Hm from PNB), 8.82 (2H, s, Ho of / pyridinium), UV (H ₂ O) λ ™ ,: 270 (ε11 5 * 70), 306 (ε7343) ηημ analysis: for C ₃₇ H ₃₈ N ₃ O ₁₀ SP · H ₂ O calc. - C 58.03% H 5.26% N 5.48% S 4.18% Found: 57.98 5.05 5.22 4.34.

10% Pd / C, H ₂

THF, ether,

buffer

COO "

To a solution of

p-nitrobenzyl-3- [2- (1- (3,5-dimethylpyridinium)) - ethylthio] -6a- [1- (R) -hydroxyethyl) -7-oxo-1-azabicyclo [3.2.0] hept- Add 2-ene-2-carboxylate-diphenyl phosphate (0.600 g, 0.80 mmol) in humid tetrahydrofuran (36 mL) to ether (36 mL), potassium phosphate monobasic sodium hydroxide buffer (0.05 M, pH 7.4, 44 ml) and 10% palladium on carbon (0.60 g), the resulting mixture hydrogenates for 1.25 h at 23 ⁰ C and 2.76 bar (40 psi), separates the organic layer and extracted with the buffer ( 2x5 ml). The layers of water are combined, filtered through a Ceiite pad. Wash with ether (40 mL) and remove traces of the organic solvents in vacuo. Then add the solution to a column (2.5 x 10.0 cm) of μ-Bondapak C-18. The column is eluted with water and the appropriate fractions are lyophilized to give 0.186 g (64%) of the title compound as a yellowish powder. IR (KBr) v _max : 3700-3100 (OH), 1760 (C = O of β-lactam), 1595 cm "'(carboxylate)

¹ H-NMR (D ₂ O) S: 1.21 (3H, d, J = 6, 3Hz, CH ₃ CHOH), 2.45 (6H, s, CH ₃ on pyridinium), 2.81 (i.e. , J = 9.2 Hz, H-4), 2.96 (d, J = 9.2 Hz, H-4), 3.22 (dd, J = 2.6 Hz, J = 6.2 Hz , H-6), 3.40 (t, J = 6.2 Hz, CH ₂ S), 3.84 (dd, J = 9.2 Hz, J = 2.6 Hz, H-5), 4 , 15 (m, CH ₃ CHOH), 4.71 (t, J = 6.2 Hz, CH ₂ N "), 8.21 (1H, s, Hp of pyridinium), 8.46 (2H, s, Ho of pyridinium) UV (H ₂ O) X _max : 279 (ε 8345), 296 (ε7714) ηημ [α] $ = + 40.7 (c 0.53, H ₂ O) τ _1/2 = 16, 9 h (measured at a concentration of 10 "" in phosphate buffer pH 7.4 at 36.8 ° C).

Example 3 Production of

hept [[2- (3-Hydroxymethylpyridinio) -ethyl] thio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo {3.2.0] - (5R, 6S) -3 -2-ene-2-carboxylate

CH ₂ OH

A. 3-Hydroxymethyl-1 - {2-mercaptoethyl) -pyridinium trifluoromethanesulfonate

CH ₂ OH

HSCH ₂ CH

CH ₂ OH

Trifluoromethanesulfonic acid (1.327 ml, 0.015 mol) is added dropwise to 3-pyridinemethanol (2.91 ml, 0.030 mol) and then ethylenesulfide (0.89 ml, 0.015 mol) is added. The resulting homogeneous mixture is heated (oil bath) for 20 h under N ₂ to 50 to 7O ⁰ C, then the reaction mixture in water (i5 ml) and extracted with CH ₂ Cl ₂ (5x5 ml). The aqueous phase is concentrated in vacuo and then added to a C ₁₈ reverse phase acid. It is eluted with water and then concentrated the appropriate fractions, giving a pale yellow oil. This material is chromatographed again, giving ein.fast colorless oil. After drying in vacuo (P ₂ O ₅ ), the product (4.50 g, 94%) is obtained as a viscous oil. IR (FHm) v _max : 3450 (st, 0 ^, 2560 (SChW ₁ SH) Cm- '

¹ H-NMR (d _e -acetone) δ: 9.10-8.05 (m, 4H, aromatic), 5.01 (t, J = 5.5 Hz, 2H, N-CH ₂ ), 4, 93 (s, 2H, -CH ₂ OH), 4.43 br.s, 1H, -OH), 3.43-3.18 (m, 2H, S-CH ₂ ), 2.34-2, 10 (m, 1H, SH).

2-carboxylate diphenylphosphate

CO ₂ PNB

CH ₂ OH

CO ₂ PNB

To a solution of p-nitrobenzyl-fSR.eSi-e-fi-fRl-hydroxyethyl J-SJ-dioxo-i-azabicyclo [s] .Ol-heptane-carboxylate (0.174 g, 0.50 mmol) in 2 mL of dry acetonitrile is added 0 ⁰ C under N ₂ diisopropylethylamine (0.096 ml, 0.55 mmol), then added dropwise to diphenylchlorophosphate (0.114 ml, 0.55 mmol) and the reaction mixture is stirred for 30 min at 0 ⁰ C. A solution of 3-hydroxymethyl-1 - (2-mercaptoethyl) -pyridinium trifluoromethanesulfonate (0.223 g, 0.70 mmol) in 0.50 ml of acetonitrile is then added, followed by diisopropylethylamine (0.122 ml, 10 mmol), keeping the mixture .30 min at 0 ° C. and then concentrate the reaction mixture in vacuo. The remaining, yellow, rubbery residue is taken up in water, adding so much acetonitrile that the rubbery product goes into solution. This solution is added to a C ₁₈ reverse phase column which is eluted with 15% acetonitrile-water. The appropriate fractions are lyophilized to give the product (0.305 g, 81%) as a beige solid.

IR (KBr) v _max : 3420 (br, OH), 1775 (β-lactam CO), 1,695 (-CO ₂ PNB) cm ^-1

¹ H-NMR (d ₆ -acetone) δ: 9.44-7.72 (m, 8H, aromatic), 7.22-6.31 (m, 1OH, diphenyl phosphate), 5.53, 5.27 ( ABq, J = 14 Hz, 2H,

benzylic), ^

5.04 (t, J = 7.4Hz, 2H, N-CH ₂ ), 4.75 (s, 2H, CH ₂ OfI), 4.5-3.1 (m, 8H), 1.21 (d, J = 6.3 Hz, 2H, CHMe). C. (SR.e

CH ₂ OH

0 \

CO ₂ PNB

To a solution of

ylat-diphenyl phosphate (0.145 g, 0.194 mmol) in 10 ml of TWF containing 5 drops of H ₂ O are added 6.0 ml of phosphate buffer (0.05 M, pH 7.4), 0.145 g of 10% palladium on Coal and 10 ml of ether. The mixture is hydrogenated (Parr) for 1 h at 40 psi, then filtered through a celite pad, the filter cake washed with a little H ₂ O and ether, the aqueous phase separated and extracted three times with ether. The aqueous solution is then cooled to 0 ° C and the pH with pH 7.4 buffer to 7.0. After removing the remaining volatiles in vacuo, the aqueous solution is applied to a C ₁₈ reverse phase column which is eluted with water. Lyophilization of the appropriate fractions gives the product (36 mg, 51%) as a light yellow solid. Further purification by reverse phase HPLC yields the pure product (31 mg, 41%) as a solid.

IR (KBr) ¹ H NMR

UV (H ₂ O)

v _max : 3300 (br, OH), 1 755 (β-lactam CO), 1590 (-CO ₂ ") cm ^-1

(D ₂ O) δ: 8.78-7.94 (m, 4H, aromatic), 4.83 (t, J = 6.0 Hz, 2H, N-CH ₂ ), 4.83 (s, 2H , CH ₂ OH), 4.16 (d of q, J = J '= 6.2 Hz, 1H, H-I'), 3.98 (d of t, J = 9.1 Hz, J '= 2.6Hz, 1H, H-5), 3.75-3.20 (m, 3H), 3.20-2.65 (m, 2H), 1.22 (d, J = 6.4Hz , 3H, CHMe)

X _max : 294 (ε 7614), 266 (ε 6936) nm

(PH 7.4; 36.8 ° C) 14,0h.

Example 4 Production of

(5R, 6S) -3- [2- (4-Hydroxymethylpyridinio) ethylthio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene 2-carboxylate

, N QV-CH ₂ OH

A. 4-hydroxymethyl-1- {2-mercaptoethyl) pyridinium trifluoromethanesulfonate

+ CF, SO, H

HS CH

CF ₃ SO ₃ "

CH ₂ OH

To a solution of 4-pyridinemethanol (1.635 g, 0.015 mol) in 10 ml of CH ₂ Cl ₂ is added dropwise under N ₂ at ⁰ ° C. trifluoromethanesulfonic acid (1.327 ml, 0.015 mol). A yellow-brown oil separates quickly. An additional equivalent of 4-pyridinemethanol (1.635 g, 0.015 mol) is then added to this mixture and the solvent removed under reduced pressure to give an oil. To this oil is added ethylene sulfide (0.891 ml, 0.015 mol) and heated the mixture obtained, homogeneous on an oil bath for 3 hours to about 6O ⁰ C. The reaction mixture then absorbs water in 15 mL and washed the aqueous solution with CH ₂ CI ₂ (5x5 ml). After removing residual organic solvent in vacuo, the aqueous solution is placed on a Cu reverse phase column. It is then eluted with water and the appropriate fractions are then concentrated to give an oil which is further dried in vacuo over P ₂ O ₅ to give the product (4.64 g, 97%) as a colorless oil. IR (FiIm) v _m : 3455 (st, OH), 2565 (schw, SH) Cm "'H-NMR (d _e -acetone) δ: 9.07, 8.18 (ABq, J = 6 , 8Hz, 4H, aromatic), 5.03 (s, 2H, CH ₂ OH), 4.96 (1, J = 6.5Hz, 2H, N-CH ₂ ), 4.09 (br.s,

1H.-OH).

P-nitrobenzyl {5R, BS) -3- [2- {4-hydroxymethylpyridinio) ethylthio] -6- [1- {R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0 ] hept-2-ene-2-carboxylate diphenylphosphate

CO ₂ PNB

CH ₂ OH

To a solution of p-nitrobenzyl- (5R, 6S) -6- [1- (R) -hydroxyethyl] -3,7-dioxo-1-azabicyclo [3.2.0] heptane-2-carboxylate (0.348 g, 1 , 0 mmol) in 5 ml of dry acetonitrile are added under N ₂ at ^{0 0} C dropwise diisopropylethylamine (0.191 ml, 1.1 mmol) and then diphenylchlorophosphate (0.228 ml, 1.1 mmol). The resulting, golden yellow solution is stirred for 40 min at ⁰ ° C, this solution is added to a solution of 4-hydroxymethyl-1- (2-mercaptoethyl) -pyridinium trifluoromethanesulfonate (0.447 g, 1.4 mmol) in 1 ml

Acetonitrile, followed by diisopropylethylamine (0.191 mL, 1.1 mmol), separating a reddish-black, gummy product from the reaction mixture, after 20 min at ⁰ ° C, filtered the reaction mixture and concentrated in vacuo. The residue is taken up in the smallest possible amount of acetonitrile-water (1: 1) and the mixture is applied to a Ci _a reverse phase column.

Elute with 25% acetonitrile-water and then lyophilize the relevant fractions to give the product (0.353 g, 47%) as a cream solid.

IR (KBr) v _max : 3240 (br, OH), 1775 (β-lactam CO), 1695 (-CO ₂ PNB) cm "'

¹ H-NMR (d _e -acetone) δ: 9.24-7.84 (m, 8H aromatic), 7.4-6.9 (m, 1H, diphenyl phosphate), 5.52, 5.24 (ABq , J = 14 Hz, 2H,

benzylic), 5.15-4.80 (m, 4H), 4.45-3.05 (m, 7H), 1.35 (d, J = 6.6Hz, 3H, CHMe). C. (5R, 6S) -3-t2- (4-hydroxymethylpyridinio) ethylthio] -6- {1- {R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2 -en-2-carboxylate

CO ₂ PNB

CH ₂ OH

CH ₂ OH

A mixture of '

p-Nitrobenzyl (5R, 6S) -3- [2- (4-hydrOxymethylpyridinio) -ethylthio3-6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2 -en-2-carboxylate-diphenyl phosphate (0.348 g, 0.465 mmol) and 10% palladium on carbon (0.35 g) in 11 mL phosphate buffer (0.05 M, pH 7.4), 5 mL THF, and 10 ml of ether is hydrogenated for 1.25 h at 2.76 bar (40 psi). Filter the mixture through a celite pad, wash the aqueous phase with ether (3x), adjust the pH of the aqueous solution to 7.0 using another pH 7.4 buffer, remove residual volatiles in vacuo and place the aqueous solution on a C ₁₈ reverse phase column. It is then eluted with 2% acetonitrile-water and then lyophilized to give a tan solid. This material is rechromatographed (C ₁₈ reverse phase / H ₂ O) to give the desired product (0.060 g, 36%) as a light yellow solid.

^ v _{ma! C} : 3400 (br, OH), 1 755 ( _? -lactam OH), 1590 (-CO ₂ -) cm- '

(D ₂ O) δ: 8.73,7.96 (ABq, J = 6.8Hz, 4H, aromat.), 4.93 (s, 2H, CH ₂ OH), 4.77 (t, J = 6.0 Hz, 2H, N-CH ₂ ), 4.15 (d of q, J = J '= 6.3 Hz, 1H, H-1'), 3.96 (d of t, J = 9.2Hz, J'-2.6Hz, 1H, H-5), 3.65-3.20 (m, 3H), 3.13-2.62 (m, 2H), 1.21 ( d, J = 6.3 Hz, 3H ₁ CHMe). , - -...

UV (H ₂ O) X _013x : 295 (ε 6880), 256 (ε 5595), 224 (ε 8111) nm τ, _{/ 2} : (pH 7.4, 36.8 ×) 14.5 h.

, ·. I

Example 5 Preparation of 3- {2- {1- (2-methylpyridinium)) - ethylthio] -6a- [1- {R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene 2-carboxylate

IR (KBr) ¹ H NMR

COO

A. 1- (2-Mefcaptoethyl) -2-metriylpyridinium methanesulfonate

+ MsOH

55 ° C, .21 h

To a suspension of 2-methylpyridinium methanesulfonate in 2-methylpyridine prepared by adding methanesulfonic acid (0.65 ml, 0.010 mol) to cold 2-methylpyridine (2.17 ml, 0.022 mol) is added ethylene sulfide (0.655 ml, 0.011 mol), the reaction mixture is stirred for 21 h under a nitrogen atmosphere at 55 ⁰ C, cooled to 23 ° C and diluted with 5 ml of water. The aqueous solution is washed with ether (6x4 ml). It removes traces of organic solvents by pumping. The solution is then placed on a column (2.5 x 10.0 cm) of μ-γοηαθρθο ^ -δδ, the column eluted with water and lyophilized the appropriate fractions to give 2.13 g (85%) of the titer compound.

IR (FiIm) v _max : 2520 (SH), 1 623 (pyridinium), 1574, 1512, 1485, 1412, 1195 (SuIfOnBt) ₁ IOSSCm- ^{1 1} H-NMR (DMSO-d _e + D ₂ O) δ: 2.37 (3H, s, CH ₃ SO ₃ -), 2.83 (3H, s, CH ₃ on pyridinium), 3.09 (2H, J = 6.9Hz, CH ₂ S), 4.71 (2H, t,

J = 6.9 Hz, CH ₂ N *), 7.93 (2H, m, Hm of pyridinium), 8.44 (1H, m, Hp of pyridinium), 8.89 (1H, m, Ho of pyridinum ) UV (H ₂ O) X _max : 266 (ε 3550) ηημ.

For example, p-nitrobenzyl-3- [2- (1- (2-methylpyridinium) -ethylthio] -6a [1- {R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept. 2-en-2-diphenyl cärboxylat

OH

1) NEt (IPr)

2) ClP (OPh) ₉ : = £

3) hS

COOPNB

4) NEt (IPr) ₂

COOPNB (PhO) ₂ PO "

To a cold (0 ⁰ C) solution of p-nitrobenzyl 6a- [1- (R) -hydroxyethyi] -3,7-dioxo-1-azabicyclo [3.2.0] heptane-2-carboxylate (0.523 g, 1 , 50 mmol) in acetonitrile (6 ml) (stored under nitrogen) are added diisopropylethylamine (0.314 ml, 1.80 mmol) followed by diphenylchlorophosphate (0.373 ml, 1.80 mmol). The reaction mixture was stirred for 30 min at ⁰ ° C and treated with a solution of 1- (2-mercaptoethyl) -2-methylpyridinium methanesulfonate (0.530 g, 2.16 mmol) in 18 ml of acetonitrile and then with diisopropylethylamine (0.314 ml, 1.8 mmol). The reaction mixture is stirred for 1 h at 0 ° C, diluted with cold (0 ⁰ C) water (26 ml) and the mixture is placed on a μ-BondapakCuj column (3.5 x 7.0 cm). The acid is eluted with 25% acetonitrile-75% water and with 50% acetonitrile-50% water. After lyophilization of the appropriate fractions 1.06 g (96%) of the titer compound are obtained as a yellowish powder

IR (KBr) v _max : 3650-3100 (OH), 1 770 (C = O of β-lactam), 1 695 and 1 690 (C = O of the PNB ester), 1 630 (pyridinium), 1 595 ( phenyl),

1 518 (NO ₂ ), 1 335 (NO ₂ ), 890 cm - '(NO ₂ ) ¹ H-NMR (DMSO-d _a ) δ: 1.15 (3H, d, J = 6.1 Hz, CH ₃ CHOH), (2.87 s, _CH3 on pyridinium), 3.6-4.4 (2H, m, H-5, CH ₃ CHOH), 4.75 (2H, m, CH ₂ N ^T) , 5.37 (center of ABq, J = 14 Hz, CH ₂ of PNB), 6.5-7.4 (10H, m, phenyl), 7.70 (2H, d, J = 8.8 Hz, Ho of PNB), 8.0 (2H, m, Hm of pyridinium), 8.24 (2H, d, J = 8.8 Hz, Hm of PNB), 8.50 (1H, m, Hp of pyridinium) , 8.95 (1H, br.d,

J = 6.1 Hz, Ho of pyridinium) UV (H ₂ O) X _max : 265 (ε 11 990), 3.14 (ε 8020) πιμ

 C. 3- [2- (1- (2-Methylpyridinium)) ethylthio] -6a [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2 ene-2-carboxylate

OH

1050 Pd / C, H _{2 v}

THF, ether, buffer

COOPNB

To a solution of,

p-nitrobenzyl-3- [2- (1- (2-methylpyridinium)) - ethylthio] -6a- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2- en-2-carboxylate-diphenyl phosphate (0.66 g, 0.90 mmol) in humid tetrahydrofuran (34 mL) is added ether (34 mL), potassium phosphate monobasic sodium hydroxide buffer (0.15 M, 16.5 mL). pH 7.22) and 10% palladium on carbon (0.66 g) and hydrogenates the resulting mixture for 1.25 h at 1.76 bar (40 psi) and 23 ^° C. The organic layer is separated and extracted with buffer (2x6 ml). The layers of water are combined, filtered through a celite pad, washed with ether (40 ml), removing traces of organic

Solvent by applying a vacuum and .u BonaapaK-u ₁₈ -sauie (ζ, ο χ iu cmj. One eiuien water and lyophilized the appropriate fractions to give 0.098 g (31%) of the title compound as a yellowish powder.

IR (KBr) v _max : 3650-3100 (OH), 1755 (C = O of β-lactam), 1630 (pyridinium), 1595 cm "'(carboxylate).

'H NMR (D ₂ O) δ: 1.20 (3H, d, J = 6.3 Hz, CH ₃ CHOH), 2.83 (s, CH ₃ on pyridinium), 2.7-3.1 (5H, H-4, CH, on pyridinium), 3.1-3.7 (3H, m, CH ₂ S, H-6), 3.90 (dd, J = 9.1 Hz, J = 2 , 6 Hz, H-5), 3.1 (m, CH ₃ CHOH), 4.78 (t, J = 6.2 Hz, CH ₂ N ^+), 7.8 (2H, m, Hm of

Pyridinium), 8.3 (1H, m, Hp of pyridinium), 8.65 (IH, s, Ho (pyridinium) UV H ₂ O) _λ ™ χ: 268 (ε9350), 196 (ε8840) Γημ

+ 41 °

15.0 h (measured at a concentration of 10 ^"1 M in phosphate buffer pH 7.4 at 36.8 ⁰ C).

Example 6 Preparation of S

COO

A. i -jZ-Mercaptoethyl J 1 -methylpyridinium methane sulphonate

· + MsOH

55 ⁰ C,

24 hours

MsO "

To a suspension of 4-picolinium methanesulfonate in 4-picoline prepared by adding methanesulfonic acid (0.65 ml, 0.010 mol) to 4-picoline (2.14 ml, 0.022 mol) is added under cooling ethylene sulfide (0.655 ml, 0.011 mol ), the reaction mixture is stirred for 24 h under a nitrogen atmosphere at 55 ⁰ C, cooled to 23 ° C and diluted with water (5 ml) and ether (10 ml). The organic layer is separated, the aqueous layer washed with ether (5 × 5 ml) and applied to a μ-Bondapak C, _r column (2.5 × 10 cm) after removing traces of ether under reduced pressure. Elute the column with a 15% acetonitrile-85% water mixture and lyophilize the appropriate fractions. This gives 2.66 g (100%) of a colorless syrup. IR (FiIm) v _max : 2500 (SH), 1640 (pyridinium), 1572.1520, 1478.1200 (sulfonate), '1040, 833 and 768 cm " ^{1 1} H-NMR (DMSO-de) δ: 2, 31 (3H, s, CH ₃ SO ₃ -), 2.62 (s, CH ₃ on pyridinium), 2.2-2.9 (4H, SH, CH ₃ on pyridinium), 3.04 (2H ₁ m , CH ₂ S),

4.68 (2H, t, J = 6.4Hz, CH ₂ N ⁺ ), 8.01 (2H, d, J = 6.6Hz ₁ Hm of pyridinium), 8.89 (2H ₁ d, J = 6.6 Hz ₁ Ho of pyridinium), UV (H ₂ O) X _max : 256 (έ 4100), 221 (ε 7544) ηημ. ,

B. 1- (2-mercaptoethyl) -4-methylpyridinium p-toluenesulfonate

. + pTsOH

PTSO "

To a suspension of p-toluenesulfonic acid (1.72 g, 0.01 mol) in benzene (6.5 ml) is added 4-picoline (1.17 ml, 0.012 mol), the resulting mixture is stirred for 30 min under a Nitrogen atmosphere at 23 ° C, treated with ethylene sulfide (0.65 ml, 0.011 mol) and stirred for 24 h at 75 ⁰ C. It gives more ethylene sulfide (0.65 ml, 0.01-1 mol), stirred for a further 24 h at 75 ° C, the reaction mixture is cooled to 23 ° C and diluted with 5 ml of water and 8 ml of ether. The aqueous layer is separated, washed with ether (3x8 ml), traces of organic solvents removed in vacuo and chromatographed on μ-Bondapak C-18 using water as eluent. 2.94 g (90%) of the titer compound are obtained as a colorless syrup.

IR (film) v _max : 2510 (SH), 1640 (pyridinium), 1595.1 582.1474.1 200 (sulfonate), 1031.1 010, 818 cm - '' H NMR (DMSO-d ₆ ) δ: 2.29 (3H, s, CH ₃ on pyridinium), 2.61 (s, CH ₃ Ph), 2.4-2.8 (4H, SH ₁ CH ₃ Ph) ₁ 3.03 ( 2H, m [addition of D ₃ O gives t, J = 6.4 Hz, at 3.04], CH ₂ S), 4.68 (2H, t, J = 6.4 Hz, CH ₂ N ⁺ ), 7.11, 7.49 (4H, 2d, J = 7.9 Hz, phenyl), 8.00 (2H, d, J = 6.5 Hz, Hm of pyridinium), 8.89 (2H ₁ d, J = 6.5 Hz, Ho of pyridinium).

UV (H ₂ O) X _max : 256 (ε 4315), 222 (έ 17045) ηημ. ' ,

C. p-Nitrobenzyl-3- [2- (1- (4-methylpyridinium)) ethylthio] -6a- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept -2-eh-2-carboxylate diphenylphosphate

NEt (iPr) ₂

OH

2) ClP (OPh) ₂

ΓΟ = °

£> - N- ^ y 4) NEt (IPr) ₂ COOPNB

(PhO) ₀ PO COOPNB ² »

To a cold (0 ⁰ C) solution of p-nitrobenzyl-p-ea-IRj hydroxyethylj-SJ-dioxo-i ^ azabicycloß .Olheptan ^ -carboxylate (0.522 6, 1.5 mmol) in acetonitrile (6 ml) (stored under nitrogen) are added diisopropylethylamine (0.314 ml, 1.8 mmol) and then diphenylchlorophosphate (0.373 ml, 1.9 mmol), the reaction mixture is stirred 45 min and treated dropwise with a solution of 1- (2-mercaptoethyl) 4-methylpyridinium methanesulfonate (0.539 mL, 2.16 mmol) in acetonitrile (1.8 mL) followed by diisopropylethylamine (0.314 mL, 1.8 mmol). Stir the reaction mixture for 1 h at ⁰ ° C, diluted with cold ⁽⁰ ° C) water (24 ml), and outputs to a μ-Bondapak C-18 column (2.5 x 8.5 cm). The column is eluted first with a 25% acetonitrile-75% water mixture (100 ml), then with a 50% acetonitrile-50% water mixture (100 ml), and after lyophilization of the appropriate fractions 0.91 g (83 %) of the title compound as a yellowish powder

IR (KBr) v _max : 3700-2800 (OH), 1770 (C = 0 of β-lactam), 1700 (C - 0 of the PNB ester), 1640 (pyridinium), 1595 (phenyl), 1520

(NO ₂ ), 1 340 (NO ₂ ), 890 cm - '(NO ₂ )' Ή-NMR (DMSO-d ₆ ) δ: 1.16 (3H, d, J = 6.2 Hz, CH ₃ CHOH ), 2.61 (s, CH ₃ on pyridinium), 3.1-3.7 (3H, m, H-6, CH ₂ S), 3.7-4.4 (2H, m, H-5 , CH ₃ CHOH), 4.79 (2h, br.t, J = 6.3 Hz, CH ₂ N-), 5.17 (d _t J - 4.9 Hz, OH), 5.37 (center of the ABq, J = 14.1 Hz, CH ₂ of PNB), 6.7-7.4 (10H, 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 ₂ O) X _max : 262 (ε 10835), 311 ( ε 9670) m μ analysis: for C ₃₆ H ₃₆ N ₃ O ₁₀ SP · 1.5H ₂ O calculated 56.84% H 5.17% N 5.52% S 4.21% found: 56.89 5, 13, 5, 19, 4:41.

D. 3- [2- (1- (4-Methylpyridinium)) ethylthio3-6a [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2 carboxylate

10% Pd / C,

THF, ether, buffer

COOPNB (PhO) ₂ PO *

- ,, · ^co ° "

To a slogan of ~

p-nitrobenzyl-3- [2- (1- (4-methylpyridinium)) - ethylthio] -6a- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2- En-2-carboxylate diphenyl phosphate (0.587 g, 0.80 mmol) in wet tetrahydrofuran (30 mL) is added with ether (30 mL), potassium phosphate monobasic sodium hydroxide buffer (0.15 M, 14.7 mL, pH 7.22 ) and 10% palladium on carbon (0.59 g), hydrogenates the resulting mixture for 1.25 h at 23 ° C under a pressure of 1.76 bar (40 psi), separates the organic layer and extracts with the Buffer (2x6 ml). The aqueous extracts are combined, filtered through a celite pad, washed with ether (3 × 20 ml), traces of organic solvents are removed by pumping and applied to a μ-Bondapak C-18 column (2.5 cm × 10 cm). Elution of the column with water and lyophilization of the appropriate fractions yields 0.136 g (49%) of the title compound as a yellowish powder. IR (KBr) v _max : 3700-3000 (OH), 1770 (C = O of β-lactam), 1642 (pyridinium), 1592 cm ^-1 (carboxylate) ¹ H-NMR (D ₂ O) δ: 1, 19 (3H, 6, J = 6.3 Hz, CH ₃ CHOH), 2.59 (3H, s, _CH3 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, CH ₂ S, H-6), 3.86 (dd, J = 9.1 Hz, 'J = 2.6 Hz, H-5), 4.12 (m, CH ₃ CHOH), 4.5-4.9 (CH ₂ N m 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),

\ _max : 256 (ε 5510), 262 (ε 5360), 296 (ε 7050) nm

+ 20.8 ° (C 0.48, H ₂ O)

12.8 h (measured at a concentration of 10 "" M in a phosphate buffer pH 7.4 at 36.8 ° C).

Example 7 Production of

(5R) -3- [2- {4-Methylthiopyridinio) ethylthio] - (6S) - [{1R) -hydro3Cyethyl] -7-oxo-1-azabicyclo {3.2.0] hept-2-ene-2 carboxylate

OH

A. A-methylthiopyridine '*'

1) MeJ / EtOH

SH _> ^ |> -SMe

2) NaGH

A. 4-Methyithiopyridine +

4-Mercaptopyridine (5.55 g, 50.0 mmol, Aldrich) is dissolved in 50 ml of boiling, absolute ethanol. The insoluble material is filtered off on Celite. The Ftitrat is heated for re-dissolving and when it is cooled to about 5O ⁰ C, is added methyl iodide (3.17 ml, 51.0 mmol; Aldrich) at a time to. The mixture is cooled to crystallize. The solid is filtered off, whereby

to obtain 6.77 g (26.7 mmol, yield 53.5%) of the title compound as the hydroiodide.

¹ H-NMR (D ₂ O) δ: 2.70 (3H, s, -SCH ₃ ) and 7.65-7.77-8.35-8.48 ppm (4H, A ₂ B ₂ type, aromat.Hs) IR (Nujol) v _max : 1615.1 585 (aromat.) and 780 cm ^-1

UV (H ₂ O) X _max : 227 (ε 2.02 x 1 fj *) and 298 nm {ε 1.64 x 10 *).

The hydroiodide (6.33 g, 25.0 mmol) is dissolved in 40 mL of water, insoluble material removed, washed with 10 mL of water, added to the filtrate at 0-5 ° NaOH pellets (5 g) and extracted with Et ₂ O (3 x 25 ml), the aqueous layer with NaC! is saturated, the combined organic extracts are washed with brine (2'x), dried (MgSO ₄ ) and concentrated to give 2.92 g (23.4 mmol, 50% overall yield) of the title compound as an oil.

¹ H-NMR (CDCl ₃ ) δ: 2.48 (3H, s, -SCH ₃ ) and 7.03-7.13-8.38-8.48 ppm (4H, A ₂ B ₂ -typ, aromat .Hs) IR (FiIm) v _max : 1580 and 800 cm " ¹ .

The preparation of this compound is described by King and Ware in J. Chem. Soc., 873 (1939). The one described there

Procedure was used

B. 4-Methyithio-N- (2-mercaptoethyi) pyridinium methanesulfpnate

CH ₃ SO ₃ H ^ ^ 'Q ^ SM «

₃₃

+ N (J) -SMe - ^ - 4 * HS ^^ N ^ y

V ^ / 50-60 °

CH, SO

j j

4-Methylthiopyridine (2.75 g, 22.0 mmol) is slowly added to methanesulfonic acid * (0.65 mL, 10.5 mmol) while cooling in an ice bath. To this solid is added ethylene sulfide ⁺ (0.66 ml, 11.0 mmol; Aldrich) and the mixture is heated 21 h at 50 to 6O ⁰ C. With the progress of reaction, the solid goes into solution. After cooling, the reaction mixture is dissolved in 5 ml of water, washed with Et ₂ O (5x4 ml), the flaky filtered. Aqueous layer over celite and the filtrate purified by reverse phase silica gel column chromatography (C ₁₈ micro Bondapack 10 g), eluting with water. Fractions of 10 ml each were collected. Fractions 2 and 3 are combined and again cleaned with the reverse phase column. Fraction 2 gives 1.258 g (4.48 mmol, yield 42.6%) of the title compound as a viscous oil. ¹ H-NMR _(DMSO-d6, CFT-20) δ: 2.32 (3H, s, MeSO ₃ '), 2.72 (3H, s, -SMe), 2.68 (1H, m, SH ), 2.9-3.2 (3H, m, -CH ₂ S-), 4.59 (2H, t,

J = 6.4Hz, -CH ₂ N "), 7.97 (2H, d, J = 7.2Hz, aromat.Hs) and 8.72ppm (2H, d, J = Hz, aromat. Hs ) IR (neat) V _018x. 1630.1200 (br, - SO ₃ "), 785 and 770 em" ¹ _x..

* These reagents were distilled before use.

C. (5R) -p-Nitrobenzyl-3- [2- (4-methylthiopyridino) -ethylthio] - (6S) - [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0 ] hept-2-ene-2-carboxylate chiorid

OH

 ο *

I) ClPO ((V).

CO ₂ PNB

_ms0

OH

SMe

CO ₂ PNB

To a solution of (5R) -p-nitrobenzyl-3,7-dioxo (6S) - [(1R) -hydroxyethyl] -1-azabicyclo [3.2.0] heptane (2R) -carboxylate (475 mg, 1 , 36 mmol) and diisopropylethylamine (0.24 ml, 1.4 mmol) in 5 ml of CH ₃ CN are added under nitrogen atmosphere at 0 to 5 ⁰ C diphenylchlorophosphate (0.29 ml, 1.41 mmol), the mixture is stirred At 0 to 5 ° C for 30 minutes, to this mixture is added an oily suspension of 4-methylthio-N- (2-mercaptoethyl) -pyridinium methanesulfonate (678 mg, 1.45 mmol, 60% purity) in 1.5 ml CH ₃ CN and then diisopropylethylamine (0.24 ml, 1.4 mmol) and the mixture is stirred for 1 h at 0 to 5 ° C. Immediately after the addition of the base, a yellow precipitate forms. The precipitate is filtered off and washed with 3 ml of cold CH ₃ CN to give 413 mg of a yellowish solid. This is triturated with 10% MeOH in water (5 ml) to give 341 mg (0.618 mmol, yield 45.4%) of the titanium compound as white crystals; Mp 118 to 120 ^° C.

¹ H-NMR (DMSO-de, CFT-20) δ: 1.16 (3H, d, J = 6.1 Hz, 1'-CH ₃ ), 2.72 (3H, s, -SCH ₃ ), 3.1-3.7 (5H, m), 3.7-4.3 (2H, m), 4.71 (2H, t, J = 6.3 Hz, -CH ₂ N ⁺ ), 5, 15 (1H, d, J = 4.9Hz, OH), 5.20-5.35-5.40-5.55 (2H, ABq, CO ₂ CH ₂ -Ar), 7.70 (2H, d, J = 8.8 Hz, nitrophenyl Hs), 7.97 (2H, d, J = 7.0 Hz, pyridinio-Hs), 8.25 (2H, d, J = 8.8 Hz, nitrophenyl -Hs) and 8.76 ppm (2H, d, J = 7.1 Hz, pyridinio-Hs)

IR (Nujol) v _max : 3250 (OH), 2775 (β-lactam), 1700 (ester) and 1625 cm ^-1 (pyridinio) UV (abs.EtOH) X _max : 308 nm (ε 4.47x10 ") ' ,

[a]% = + 24.8 ° (c 0.5, MeOH) Analysis: for C ₂₄ H ₂₆ N ₃ O ₃ S ₂ Cl · H ₂ O calculated 50.56% H 4.95% N 7.37 % found: 50.63 4.72 6.89.

D. (5R) -3- {2- (4-Methylthiopyridinio) ethylthio] - {6S) - [{1R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene 2-carboxylate

H ₂ / Pd-C

PH 7.4 buffer

CO ₂ PNB

(5R) -p-nitrobenzyl-3- [2- (4-methylthiopyridinio) ethyl-thio] - (6S) - [(1R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] -hept 2-ene-2-carboxylate chloride (380 mg, 0.688 mmol) is dissolved in 31.5 ml of THF and a pH 7.40 phosphate buffer (31.5 ml, 0.05 M, Fisher) diluted with Et ₂ O (31.5 mL), mix this solution with 10% Pd-C (380 mg, Engelhard) and hydrogenate for 1 h at room temperature with a Parr shaker at 2.4 bar (35 psi). The aqueous layer is filtered through Celite to remove the catalyst and the celite pad washed with water (2x5 mL). The filtrate and washings are combined and washed with Et ₂ O (2 x 30 mL). The aqueous layer was removed under reduced pressure of organic solvents and purified by reverse phase column chromatography (C ₁₈ -Microbondapak, 13 g; Waters Associates), eluting with water. Fractions with UV absorption at 307 nm are collected (about 1 liter) and lyophilized to give 127 mg (0.334 mmol, yield 48.5%) of the titanium compound as a yellowish powder

receives

¹ H-NMR

(D ₂ O, CFT-20) δ: 1.20 (3H, d, J = 6.4 Hz. 1'-CH ₃ ), 2.64 (3H, s, -SCH ₃ ), 2.81 ( 2H, m, -SCH ₂ -), 3.19 (1H, dd, J ₆ -, = 6.1Hz, J ₈ -S = 2.6Hz, 6-H), 3.32 (2H, dd , J = 11 Hz, J = 5.5 Hz, 4-Hs), 3.92 (1H, dt, J = 9.2 Hz, J ₅ ^ = 2.6 Hz, 5-H), 4.1 (1H, m, 1'-H), 4.61 (2H, t, J = 5.9Hz, -CH ₂ N ⁺ ), 7.70 (2H, d, J = 7.1Hz, aromat. Hs) and 8.40 ppm (2H, d, J = 7.1 Hz, aromat.Hs) (KBr, Pill) v _max : 3400 (OH), 1 750 (β-lactam), 1630 (pyridinium) and 1590 cm " ¹ (carboxylate) X _max : 231 (ε 9800) and 307 nm (ε 25000) '+ 3.14 ° (c 0.5, H ₃ O).

Example 8 Preparation of

OMe

A. 1- (2-Mercaptoethyl) -3-methoxypyridinium methanesulfonate

MeO

MSO "

MeO

MsOH

To pre-cooled (5 ° C) 3-methoxypyridine (698 mg, 6.4 mmol) is added dropwise methanesulfonic acid (0.216 ml, 3.05 mmol) and

Ethylene sulfide (0.19 ml, 3.2 mmol). The mixture is then heated for 18 h to 6O ⁰ C, cooled to 20 ⁰ C, diluted with water (10 ml) and washed with ether (3 x 10 ml). The aqueous phase is treated for 15 minutes at high vacuum and gives on a Ci8 reverse phase column. The title compound is eluted with water. Combine the appropriate fractions and concentrate under high vacuum to give the desired thiotone (61.6 mg, 76.3% yield).

IR (CH ₂ Cl ₂ ) v _max : 2550 (schw, SH) and 1 620.1600, 1585 cm ^-1 (m, aromat.)

¹ H-NMR (DMS0-d _e) δ: 8.90 to 7.90 (4H, m, aromat.CH), 4.72 (2H, t _t J = 6.6 Hz, CH ₂ N ^+), 4.01 (CH, s, OCH ₃ ), 3.5-3.0 (m,

hidden), 2.66 (1H, dd, J = 9.5 Hz, J = 7.5 Hz, SH) and 2.31 ppm (3H ₁ S ₁ CH ₃ SO ₃ -). B. l

2-carboxylate

1) EtN (- <) ₂ 2) ClPO (0 ^) ₂

HS

.0Me

4) EtN (- <) ₂

OMe

CO ₂ PNB

CO ₂ PNB

A cold (0 ⁰ C) solution of p-nitrobenzyl 6a- [1 '- (R) -hydroxyethyl] -3,7-dioxo-1-azabicyclo [3.2.0] heptane-2-carboxylate (1.04 g , 3 mmol) in 12 ml of acetonitrile are treated dropwise with diisopropylethylamine (0.63 ml, 3.6 mmol) and diphenylchlorophosphate (0.75 ml, 3.6 mmol) and stirred for 30 min at ⁰ ° C. The resulting enol phosphate is treated one with

1- (2-Mercaptoethyl) -3-methoxypyridinium methanesulfonate (. 1.14 g, 4.30 mmol) in 7 mL of CH ₃ CN, 0.63 mL (4.30 mmol) of diisopropylethylamine, stirred for 30 min and cooled to 30 min precipitated at -10 ⁰ C. the solid was filtered from the mixture, washed with cold acetonitrile (2 ml) and dried to afford the title compound is obtained (1.32 g, yield 82%). IR (Nujol) v _max : 3320 (m, OH), 1 780.1 765 (st ,, β-lactam (C =), 1700, 1695 (m, ester C = 0) and 1520 cm ^-1 (st, NO ₂ ) ¹ H NMR (DMSO-d _e ) 5: 9.01 (1H, bs, H-3 aromat), 8.75 (1H, bd, J = 5.4 Hz, H-6 aromat.), 8.35-7.95 (4H, m, H-aromat), 7.70 (2H, d, J = 7.7Hz, H-aromat), 5.37 (2H, center of ABq, J = 13 Hz, _CH2 PNB), 5.17 (1H, d, J = 4.9 Hz, OH), 4.87 (2H, t, J = 6.3 Hz, CH ₂ -N ⁺ ), 4.35-3.75 (2H, m, H-5 and H-1 '), 4.00 (3H, s, OCH ₃ ), 3.56 (part of t, J = 6.3 Hz , CH ₂ S), 3.5-3.20 (3H, m, H-6, H-3) and 1.16 ppm (3H, d, J = 6.1 Hz, CH ₃ CHO).

Cl "OMe

H,

CO ₂ PNB

A solution from

Ylate (600 mg, 1.12 mmol) in THF (25 mL), ether (25 mL) and pH 7.4 phosphate buffer (0.1 M, 25 mL) is hydrogenated in a Parr shaker at 2.76 for 1 h bar (40 psi) over 10% Pd / C. The mixture is diluted with ether and the aqueous phase is filtered through a No. 52 hardened filter paper. Wash the aqueous layer with ether (2 x 20 mL), apply a vacuum, and place on a silica gel reverse phase column. The title compound is washed with water containing 2% and 5% acetonitrile. The appropriate fractions are combined and lyophilized to give a yellow solid which is purified by HPLC to give the penem carboxylate (150 mg, 38%).

IR (Nujol) v _max : 1 750 (st, β-lactam C = 0) and 1 580 cm - '(st, carboxylate)

¹ H-NMR (P ₂ O) δ: 8.55-8.30 (2H, m, H-2, H-6 aromat.), 8.17-7.75 (2H, m, H-3, H-4 aromat.), 4.77 (2H, t, J = 5.9 Hz, CH ₂ N ⁺ ), 4.10 (1H, part of 5 lines, J = 6.3 Hz, HT), 3 , 97 (3H, s, OCH ₃ ), 3.85, 3.82 (2 lines, part of dt, J = 2.6 Hz, part of H-5), 3.42 (2H, t, J = 5.9 Hz, CH ₂ -S), 3.25 (1H, dd, J = 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 = 6.4Hz, CH ₃₎

UV (H ₂ O, c 0.05) X _max : 290 (e 10517), 223 (ε 6643),

τ _1/2 (0.1 M pH 7.4 phosphate buffer, 37 ^° C.) = 20 h.

Example 9 Production of

(5R, 6S) -3- [2- (3-Methylthiopyridinio) ethylthio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicycio- [3.2.0] hept-2-ene -2-carboxylate

SMe

A. 3-Methylthio-1- (2-mercaptoethyl) -pyridinium chloride

SMe

SMe

+ HCl +

4 HSCH ₂ CH ₂ N

Cl

To a solution of 3-methylthiopyridine (2.00 g, 0.016 mol) (prepared by the method of JA Zoltewiez and C. Nisi, J. Org. Chem. 34, 765, 1996) in 10 ml of ether is added 15 ml of 1N HCI, shake the mixture thoroughly, the aqueous phase is separated off, washed with 10 ml of ether and then concentrated. The remaining hydrochloride is then dried in vacuo (P ₂ O ₅ ) to give a white solid. To this hydrochloride solid is added 3-methylthiopyridine (1.88 g, 0.015 mol) and ethylene sulfide (0.89 ml, 0.015 mol) and the resulting mixture heated (oil bath) for 15 h under N ₂ ^O 55 to 65 C. A slightly cloudy oil is obtained, which is taken up in 125 ml of water and washed with CH ₂ Cl ₂ . The aqueous solution is concentrated to about 25 ml, then gives some

UIIUI Il CU ₁ Ulli UIC IVMUIsI IUI IIWI liuycii Ä.U admonish, ijuLU ic σι 11 al (.Clic ?, νναιοι * yc i * uouiiy au ι duo wjg-wn in. * Stii ynu ^ iiduuiu,

eluted with water and then concentrated the relevant fractions to give the product as a pale yellow, viscous oil (2.66 g, 80%).

IR (FiIm) v _max : 2410 (br, -SH) cm " ¹

¹ H-NMR (de-DMSO + D ₂ O) δ: 8.88-7.88 (m, 4H, aromat), 4.70 (t, J = 6.5 Hz, 2H, N-CH ₂ ) , 3.08 (oblique t, J = 6.5 Hz, 2H, S-CH ₂ ), 2.64 (s, 3H ₁ S-Me).

B. p-Nitrobenzyl- (5R, 6S) -3- [2- (3-methylthiopyridinio) -ethylthio3-6- [1 - (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate hydrochloride

CO ₂ PNB

CO ₂ PNB

A solution of p-nitrobenzyl- (5R, 6S) -6- [1- (R) -hydroxyethyl] -3,7-dioxo-1-azabicyclo [3.2.0] heptane-2-carboxylate (0.522 g, 1, 50 mmol) in 7 ml of dry acetonitrile is cooled to ⁰ ° C., then diisopropylethylamine (0.287 ml, 1.65 mmol) is added dropwise, diphenyichlorophosphate (0.342 ml, 1.65 mmol) is added dropwise to the resulting tan solution the reaction mixture 30 min at 0 ⁰ C. It then Diisopropylethyiamin (0.313 ml, 1.80 mmol) was added and then a solution of 3-methylthio-1- (2-mercaptoethyi) -pyridiriiumch! ORID (0.398 g, 1.80 mmol) in 0.70 ml of dry DMF About 1 minute after completion of the addition, a precipitate separates from the reaction mixture. Upon further cooling to -10 ⁰ C for 10 min gives a solid, orange mass. This solid is then triturated with acetonitrile and the residue is filtered off. The residue is washed with acetonitrile, then with acetone and finally dried in vacuo to give the product (0.455 g, 55%) as a cream-colored dye. The combined filtrate is concentrated to give a yellow oil, which is taken up in as little as possible acetonitrile and cooled to 0 ° C for 30 min. This mixture is filtered, giving another 0.139 g of the product as a pale yellow solid. The total yield is 0.594 g (72%)

IR (KBr) v _max 3345 (br, -OH), 1770 (β-lactam CO), '1 680 (-CO ₂ PNB) cm ^{-1 1} H NMR (d _e -DMSO) δ: 8.98 7.96 (m, 4H, pyridinium aromat,), 8.20-7.65 (ABq, J = 7.0 Hz, 4H, PNB aromat.) 5.53-4.80 (m, 4H),

4.3-3.7 (m, 2H), 3.6-3.25 (m, 6H), 2.66 (s, 3H, S-Me), 1.16 (d, J = 6.0 Hz, 3H, CHMe). C. (5R, 6S) -3- [2- (3-l-ethylthiopyridinio) -ethylthio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2 ene-2-carboxylate

SMe

CO ₂ PNB

To a mixture of

p-Nitrobenzyl (5R, 6S) -3- [2- (3-methylthiopyridinio) ethylthio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] -hept 2-ene-2-carboxylate chloride (0.551 g, 1.0 mmol) and 10% palladium on carbon (0.55 g) in 25 ml of phosphate buffer (0.05 M, pH 7.4) are added 5 ml of THF and 25 ml of ether. The mixture is hydrogenated (Parr) for 1 h at 2.76 bar (40 psi). The reaction mixture is then filtered through Celite and the filter cake is washed with water and ether. The aqueous phase is separated, washed with additional ether (3x), removed

residual organic solvent in vacuo and the aqueous solution is cooled to ^{0 0} C and the pH with saturated, aqueous NaHCO ₃ to 7.0. This solution is then added immediately to a Ci ₈ reverse phase column, eluted with water and then lyophilized the appropriate fractions to give 0.25 g of a light-colored solid. This material is again purified by reverse phase HPLC to give the product (0.210 g, 55%) as a light yellow solid.

IR (KBr) v _max : 3400 (br, -OH), 1 755 (β-lactam CO), 1 590 (-CO ₂ ') cm ^-1

¹ H-NMR (D ₂ O) δ: 8.60-7.76 (m, 4H, arom.), 4.76 (t, J = 5.8 Hz, 2H, N-CH ₂ ), 4, 13 (d of q, J = J '= 6.3 Hz, 1H, H-1'), 3.95 (d of t,

J = 9.0 Hz, J '= 2.8 Hz, 1H, H-5), 3.45-2.75 (m, 5H), 2.59 (s, 3H, S-Me), 1, 20 (d, J = 6.4 Hz, 3H, CHMe) UV (H ₂ O): X _max : 296 (e8509), 273 (ε 13005), 231 (e11576) nm τι / 2 (pH 7.4, 36.8 ⁰ C) = 20 h.

Example 10 Production of

3- [2- (1- (2,6-dimethylpyridinium)) - ethylthio] -6a- (1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene -2-carboxylate

OH

COO

A. 1- (2-Mercaptoethyl) -2,6-dimethylpyridinium methane suberate

+ MsOH

100 ⁰ C, 42 h

-►HS

A mixture of 2,6-dimethyipyridine (19.2 mL, 0.165 mol) and methanesulfonic acid (3.27 mL, 0.050 mol) is stirred for 15 min, treated with ethylene sulfide (4.17 mL, 0.070 mol) and stirred for 42 h Nitrogen atmosphere at 100 ⁰ C. After cooling to 25 ⁰ C diluted the reaction mixture with ether (45 ml) and water (30 ml), separates the two layers and the organic layer is extracted with water (2x5 ml). The aqueous layers are combined, filtered through a celite pad, washed with ether (2 x 15 ml), organic solvent removed in vacuo, and poured onto a μ-Bondapack C-18 column (3.0 x 12 cm). Elute with a 3% acetonitrile-97% water mixture and lyophilize the appropriate fractions to give 2.5 g of the impure title compound as a syrup. Get this syrup. This syrup is again purified by HPLC (μ-Bondapak C-18) to give 0.90 g (7%) of the title compound.

IR (FiIm) v _max : 2520 (SH), 1640 and V625 (pyridinium), 1585.1490, 1200 cm- ¹ (sulfonate) ν

¹ H-NMR (DMSO-d ₆ + D ₂ O) δ: 2.36 (3H, s, CH ₃ SO ₃ -), 4.62 (2H, m, CH ₂ N ^), 7.74 (2H , m, Hm of pyridinium), 8.24 (1H, m, Hp of pyridinium)

UV (H ₂ O) X _max : 272 (ε 4080) m μ

B. p-Nitrobenzyl-3- [2- (1- (2,6-dimethylpyridinium)) ethylthio] -6a- [1- (R) -hydroxyethyl-3-oxo-1-azabicyclo- [3.2.0] hept-2-ene-2-carboxylate diphenylphosphate

1) NEt (iPr)

4) NEt (iPr).

COOPNB

COOPNB (PhO) ₂ PO

To a cold (u ~ u) solution of (j-muijuoiii.yi-u ^. ^ ,,, ,,, ...- ",, -, - ·

1.89 mmol) in 6 mL of acetonitrile (stored under nitrogen atmosphere) are added diisopropylethylamine (0.394 mL, 2.26 mmol) and diphenylchlorophosphate (0.468 mL, 2.26 mmol), the reaction mixture is stirred for 30 min and treated with a solution of 1 - (2-Mercaptoethyl) -2,6-dimethylpyridinium methanesulfonate (0.720 g, 2.73 mmol) in 3 mL of acetonitrile followed by diisopropylethylamine (0.394 mL, 2.26 mmol). The reaction mixture is stirred for 2 h at 0 ⁰ C, diluted with cold (0 ⁰ C) water (27 ml) and poured onto a μ-Bondapak C-18 column (2.5 x 9.0 cm). Elute with an acetonitrile-water mixture and lyophilize the appropriate fractions to give 0.92 g (65%) of the title compound

IR (KBr) v _{may <} : 3700-3000 (OH), 1765 (C = 0 of β-lactam), 1690 (C = O of the PNB ester), 1620 (pyridinium), 1590 (phenyl), 1517

(NO ₂ ) JSSO (NO ₂ I ₁ SeOCm- ⁵ NO ₂ ) ¹ H-NMR (DMSO-de) δ: 1.15 (3H, d, J = 6.2 Hz, CH ₃ CHOH), 2.7 -3.7 (11H, CH ₂ S, 2-CH ₃ on pyridinium, H-4, H-6), 3.7-4.4 (2H, CH ₃ CHOH, H-5), 4.7 ( 2H, m, CH ₂ N ^+), 5.14 (1H, d, J = 4.5 Hz, OH), 5.37 (center of ABq, J = 13.2 Hz, _CH2 of PNB), 6 , 7-7.5 (10H, m, phenyl), 7.5-8.7 (7H, pyridinium, H of PNB)

UV (H ₂ O) X _ma ": 274 (ε 14150), 319 (ε 9445) m μ ·

1 0% Pd / C, H?

THF, ether, buffer

COOPNB

goo

To a solution of

phenylphospha-p (0.80 g, 1.07 mmol) in humid tetrahydrofuran (42 mL) are added ether (42 mL), potassium phosphate monobasic sodium hydroxide buffer (0.15 M, pH 7.22, 21 mL) and Charge 10% palladium on carbon (0.80 g), hydrogenate the resulting mixture for 1 h at 23 ° C and a pressure of 40 psi and filter through a pad of Celite. Separate the two layers and extract the organic layer with the buffer (3 x 8 ml). The aqueous phases are combined, washed with 50 ml of ether, traces of organic solvents are removed in vacuo and applied to a μ-Bondapak C-18 column (3.0 × 10.2 cm). The column is eluted with a 5% acetonitrile-95% water mixture and the appropriate fractions are lyophilized to give 0.246 g (63%) of the title compound as a yellow powder.

v _max : 3700-2800 (OH), 1750 (C = O of β-lactam), 1 620 (pyridinium), 1585 cm ^-1 (carboxylate) (D ₂ O) δ: 1.23 (3H, d, J = 6.4Hz, CH ₃ CHOH), 2.5-3.5 (11H, H-4, H-6, CH ₂ S, 2CH ₃ on pyridinium), 3.8-4.4 (2H, CH ₃ CHOH, H-5), 4.5-4.9 (CH ₂ N ⁺ , HOD), 7.64 and 7.74 (2H, Α portion of the A ₂ B system, Hm of pyridinium), 8.07, 8.16, 8.18 and 8.27

(1H, B part of the A ₂ B system, Hp of pyridinium)

X _max : 277 (ε 9733), 300 (ε 8271) m μ:. . ' , .

+ 50.7 ° (C 0.48, H ₂ O) VfUrC ₁₃ H ₂₂ N ₂ O ₄ SI ₁ BH ₂ O. , ,

C 55.51% H 6,47% N 7,19% 55,14 _ 6,23 6,46.

IR (KBr) ¹ H NMR

UV (H ₂ O) [α] § = analysis: calculated: found:

Example 11 Production of

{5R, 6S) -3- [2- (2-methylthio-3-methylimidazolio) ethylthio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept- 2-ene-2-carboxylate

A. 2-Methylthio-3-methyl-1- (2-mercaptoethyl) imidazolium trifluoromethanesulfonate

CF ₃ Sun ₃ H y.

SMe

ISRC. CH N ..N-Ms

CF ^ SO

Trifluoromethanesulfonic acid (1.38 ml, 0.015 mol) is added dropwise to 2-methylthio-1-methylimidazole (4.0 g, 0.03 mol) (prepared as described by A. Wohl and W. Marckwald in Chem. Ber. 22 , described 1353 (1889)) at 0 ⁰ C under N _2, then ethylene sulfide (0.9 ml, 0.015 mol) to the mixture under N ₂ 24 heats h at 55 ⁰ C, the reaction mixture with ether, triturated (3X ) and the residue is taken up in acetone. Then filtered and evaporated. This gives the product (4.2 g, 82%) as a semi-crystalline solid, which is used without further purification.

IR (FiIm) λ: 2550 (black, Sch.) Cm " ¹

¹ H-NMR (d _r acetone) δ: 7.97 (s, 2H), 4.66 (t, J = 7Hz, 2H, methylene), 4.17 (s, 3H, N-Me), 3 , 20 (d of t, J = 7Hz, J '= 9Hz, 2H, methylene), 2.72 (s, (3H, S-Me), 2.20 (t, J = 9Hz, 1H, -SH).

P-nitrobenzyl- (5R, 6S) -3- [2- (2-methylthio-3-methylimidazolio) ethithio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo ( 3.2.0] hept-2-ene-2-carboxylate diphenylphosphate

SMe

-Me

To a solution of p -nitrobenzyl-fSR.eSJ-e-fi-iRI-hydroxyethylj-SJ-dioxo-i-azabicyclosc-O-heptanecarboxylate (1.40 g, 4.0 mmol) in 50 mL of dry acetonitrile one under N ₂ was added dropwise at 0 ⁰ C diisopropylethylamine (0.76 mL, 4.4 mmol) and then diphenyl chlorophosphate (0.91 mL, 4.1 mmol), stirred the reaction mixture for 1 h at room temperature, diisopropylethylamine gives (0.76 ml, 4.4 mmol) and then a solution of

2-Methylthio-3-methyl-1- (2-mercaptoethyl) imidazolium trifluoromethanesulfonate (2.0 g, 5.9 mmol) in 5 mL acetonitrile dropwise, the reaction mixture is kept at room temperature for 1.5 h and then concentrated in water Vacuum to obtain a rubbery product. This gummy product is taken up in water and placed on a C ₁₈ reverse phase column. It is then washed with water, then 20% acetonitrile-water and finally 30% acetonitrile-water. Then the appropriate fractions are lyophilized and the product (0.90 g, 30%) as a pale yellow solid

? IR (KBr) w: 3380 (br, OH), 1770 (.beta.-lactam CO) cm ^{"1 1} H-NMR (d ₆ -acetone) 5: 8.35 (brs, 1H, 8.24, 7 , 78 (ABq, J = 8.8 Hz, 4H, aromat.), 7.89 (brs, 1H), 7.25-6.91 (m, 10H, diphenyl phosphate), 5.50, 5.25 ( ABq, J = 12Hz, 2H, benzyl.), 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. (5R, 6S) -3- {2- {2-Methylthio-3-methylimidazolio) ethylthioi-6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0} hept-2-ene-2-carboxylate

To a solution of '

p-Nitrobenzyl (5R, 6S) -3- [2- (2-methylthio-3-methylimidazolio) ethylthio] -6-t1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0 ] hept-2-ene-2-carboxylate diphenyl phosphate (1.20 g, 1.56 mmol) in a mixture of 70 ml of THF, 70 ml of ether and 31 ml of phosphate buffer (0.05 M, pH 7.4 Add 1.2 g of 10% palladium on carbon, hydrogenate the mixture (Parr) for 55 min at a pressure of 2.41 bar (35 psi), filter the reaction mixture through celite and wash the filter cake with water and ether , The aqueous phase is separated, cooled to 0 ⁰ C and the pH with saturated, aqueous NaHCO ₃ to 7.0. After removal of residual organic solvents in vacuo, the aqueous solution is applied to a Cig reverse phase column. It is eluted with water and then with 8% acetonitrile-water and then the appropriate fractions are lyophilized to give 0.25 g of a solid. This material is again purified by reverse phase HPLC to give the product (0.114 g, 19%) as an off-white solid.

v _max : 3420 (OH), 1750 (β-lactam CO), 1590 (-CO ₂ -) cm " ¹ '."

(D ₂ O): δ: 7.58 (s, 2H), 4.52 (t, J = 6Hz, 2H), 4.28-3.82 (m, 2H), 3.90 (s, 3H, N-Me), 3.40-2.87 (m, 5H), 2.40 (s, 3H, S-Me),

1.20 (d, J = 6.4 Hz, 3H, -CHMe)

X _m ": 297 (ε 7572), 262 (ε 6259), 222 (ε 7955) nm.

IR (KBr) ¹ H NMR

UV (H ₂ O)

Example 12 Preparation of (5R,

A. 3-Amino-1- (2-mercaptoethyl) -pyridinium chloride

HSCH ₂ CH ₂

.HCl

3-Aminopyridine (1.50 g, 0.016 mol) is taken up in 15 ml of 1N methanolic HCl, the resulting solution is evaporated off to give the hydrochloride as an oil, 3-aminopyridine (1.32 g, 0.015 mol) and ethylene sulfide ( 0.89 ml, 0.015 mol) to this oil and the resulting mixture (oil bath) heated under N ₂ at 60 to 65 ° C for 2 h. Another equivalent of ethylene suicide (0.89 ml, 0.015 mol) is added and the mixture is heated for a further 65 h at 55 to 65 ^° C. The reaction mixture is washed with CH ₂ Cl ₂ and then taken up in 25 ml of water.

The aqueous solution is added to a Cie reverse phase column which is eluted with water. After evaporation of the relevant fractions, the product (1.26 g, 44%) is obtained as a colorless, viscous oil.

IR (FiIm) v _max : 3180 (NH ₂ ) cm- '·

¹ HNMR (d _e -DMSO) δ: 8.19-7.59 (m, 4H, aromatic J., 4.59 (t, J = 6.2 Hz, 2H, N-CH ₂ ), 3.5 (brs, 2H, -NH ₂ ), 3.20-2.77 (m, 3H).

ylat diphenyl

CO ₂ PNB

CO ₂ PNB

To a solution of p-nitrobenzyl- (5R, 6S) -6- [1- (R) -hydroxyethyl] -3,7-dioxo-1-azabicyclo [3.2.0] heptane-2-carboxylate (0.696 g, 2 , 0 mmol) in 10 ml of dry acetonitrile are added under N ₂ at 0 ⁰ C dropwise diisopropylethylamine (0.382 ml, 2.2 mmol) and

then diphenyl dichlorophosphate (0.457 mL, 2.2 mmol), stirred for 30 min at 0 ° C, add a solution of 3-amino-1- (2-mercapto-ethyl) -pyridinium chloride (0.475 g, 2.5 mmol) in 1 ml of dry DMF and then additional diisopropylethylamine (0.435 ml, 2.5 mmol), the reaction mixture is kept for 1.5 h at 0 ° C and then concentrated in vacuo. The resulting gummy product is taken up in acetonitrile-water (1: 1) and added to a C ₁₈ reversed-phase column. It is then eluted with water, then with 20% acetonitrile-water, and then the relevant fractions are lyophilized to give the product (0.730 g, 50%) as a beige solid.

IR (KBr) v: 3330 (br, OH), 3180 (br, NH ₂ ), 1770 (β-lactam CO), 1690 (-CO ₂ PNB) cm " ¹ H-NMR (d ₆ -DMSO) δ : 8.29-7.63 (m, 8H, aromat.), 7.2-6.7 (m, 10H, diphenyl phosphate), 5.47, 5.18 (ABq, J = 14Hz, 2H, benzyl .)

4.73-4.45 (m, 3H), 4.2-3.8 (m, 1H), 3.6-2.6 (m, 8H), 1.1.5 (d, J = 6, 2 Hz, 3H), CHMe. C. (5R, 6S) -3- {2- {3-aminopyridinio} -ethylthio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.03hept-2-ene] 2-carboxylate

To a mixture of

p-Nitrobenzyl (5R, 6S) -3- [2- {3-aminopyridinio) ethylthio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept- Add 2-ene-2-carboxylate-diphenyl phosphate (0.730 g, 1.0 mmol) and 10% palladium on silica (0.7 g) in 25 mL of a phosphate buffer (0.05 M, pH 7.4) 8 ml of THF and 20 ml of ether, the mixture is then hydrogenated for 1 h at a pressure of 2,76 bar (40 psi) (Parr), the resulting mixture is filtered through a pad of Celite and the filter cake is washed with water and ether. The aqueous phase is separated off, washed twice with ether and then removed residual volatiles in vacuo. The aqueous solution is immediately added to a C ₁₈ reverse phase column, which is eluted with water. Lyophilization of the relevant fractions gives 0.45 g of an off-white solid. This material is again purified by reverse phase HPLC to give the desired product (0.123 g, 35%) as an off-white solid.

IR (KBr) v _max : 3340 (br), 1 750 (br, β-lactam CO), 1 580 (br, -CO ₂ -) cm "'

¹ H-NMR (D ₂ O) δ: 8.07-7.59 (m, 4H, aromat), 4.61 (t, J = 5.8 Hz, 2H, N-CH ₂ ), 4.14 (d of q, J = J '= 6.3 Hz, 1H, HT), 3.97 (d of t, J = 9.2 Hz, J' = 2.6 Hz, 1H, H-5), 3.38 (t, J = 5.8 Hz, 2H, S-CH ₂ ), 3.24 (d of d, J = 6.0 Hz, J '= 2.6 Hz, 1H, H-6) , 3,17-2,57

(m, 2H, H-4), 1.21 (d, J = 6.3 Hz, 3H, CHMe) UV (H ₂ O) X _max : 299 (ε 7949), 256 (ε 8822) nm τ _{1 / 2} pH 7.4.36.8 ° C) = 18.5 h. '

Example 13 Production of

{5R, 6S) -3- (1- {S) -methyl-2- (1-pyridinium) ethylthio] -6- {1- {R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2. 0] hept-2-ene-2-carboxylate

COCF

and

(5R, BS) -3- [1- (R) -methyl-2- (1-pyridinium) ethylthio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo! O [ 3.2.0] hept-2-ene-2-carboxylate OH.

A.

CCO

A. dl-1- (2-Mercapto-2-methylethyl) pyridinium methanesulfonate dl-1- (2-mercapto-1-methylethyl-pyridinium methanesulfonate

SH

MsOH

Methanesulfonic acid (1.95 ml, 0.030 mol) is added slowly to cold pyridine (7.83 ml, 0.097 mol), the resulting mixture stirred for 15 min at 40 ° C, treated with dl-propylene sulfide (2.59 ml, 0.033 mol ) and stirred for 90 h under a nitrogen atmosphere at 60 ⁰ C. The pyridine is removed in vacuo, the residue is mixed with water and purified by chromatography (HPLC, Preparative Bondapak C-18). The appropriate fractions are combined and lyophilized to give dl-i- (2-mercapto-2-methylethyl) pyridinium methanesulfonate (1.14 g, 15%) as a colorless syrup.

IR (FiIm) y _max : 2520 (SH), 1640 (pyridinium), 1180 (st, CH 3 SO 3 -), 1040 (CH 3 SO 3) cm ^{-1 1} H-NMR: (DMSO-de) δ: 1.35 (d , J = 6.8 Hz, 3H, CH ₃ CHS), 2.30 (s, 3H, CH ₃ SO ₃ -), 2.90 (d, J = 8.5 Hz, 1H, SH), 3, 2-3.7 (m, CHSH), 4.52 (dd, J _{g (5m} = 12.9 Hz, J = 8.4 Hz, CHCH ₂ N *), 4.87 (dd, J _aem = 12 , 9 Hz, J = 6.0 Hz, CHCH ₂ N ⁺ ), 8.0-8.4 (m, 2H, Hm of pyridinium), 8.5-8.8 (m, 1H, Hp of pyridinium) , 9.04 (dd, J = 1.4 Hz, J = 6.7 Hz, 2H, Ho of pyridinium)

X _mflx : 208 (ε 5267), 259 (ε 3338) for C ₉ H ₁₅ NO ₃ S ₂ .2H ₂ O.

H 6,71% N 4,91% S 22,47% 6,85 4,86 22,09

UV (H ₂ O) analysis:

calculated: C 37.88% found: 37.49

Further, dl-1- (2-mercapto-1-methylethyl) -pyridinium methanesulfonate (0.82 g, 11%) is obtained as a colorless syrup.

IR (film) 'H-NMR

v _max : 2500 (SH), 1628 (pyridinium), 1180 (sulfonate), 1035 (sulfonate) cm " ¹ '

(DMSO-d ₆ ) δ: 1.69 (d, J = 6.8 Hz, 3H, CH ₃ CHN ⁺ ), 2.31 (s, 3H, CH ₃ SO ₃ -), 3.0-3, 3 (m, 2H, CH ₂ S), 4.2-5.2 (m, 1H, CHN ⁺ ) 8.0-8.4 (m, 2H, Hm of pyridinium), 8.5-8.8 (m, 1H, Hp of pyridinium), 9.0-9.2 (m, 2H, Ho of pyridinium)

UV (H ₂ O) X _max : 209 (ε 4987), 258 (ε 3838) Analysis: for C ₉ H ₁₆ NO ₃ S ₂ .1.5H ₂ O.

Calculated: C 39.11% H 6.56% N 5.07%

found ': 39.13 5.92 5.20

B. (5R, 6S) -p-nitrobenzyi-3 {1- {R, S) -methyl-2- (1-pyridinium) -ethylthio] -6- [1- (R) -hydroxyethyl] -7-oxo -1-a2abrcyclo [3.2.0] hept- ^ -ene ^ -carboxylate diphenyl phosphate

 1).

2) ClP (OPh)

COOPNB

3)

'MsO

4) -.NEt (IPt).

COOPNB

To a cold (O ⁰ C) solution of (5R, 6S) -

(0.523 g, 1.5 mmol) in 6 mL of acetonitrile (kept under nitrogen atmosphere) are added diisopropylethylamine (0.314 mL, 1.8 mmol) followed by diphenylchlorophosphate (0.373 mL, 1.8 mmol), the reaction mixture is stirred for 30 min and treated with a solution of dl-1- (2-mercapto-2-methylethyl) -pyridinium methanesulfonate (0.539 g, 2.16 mmol) in 2 mL of acetonitrile and diisopropylethylamine (0.314 mL, 1.8 mmol). The reaction mixture is stirred for 1 h at ^{0 0} C, diluted with cold (0 ⁰ C) water (24 ml) and chromatographies on a prep. Bondapak C-18 column (2.5 x 8.5 cm) using 25% to 50% acetonitrile in water as the eluent. After lyophilization, 1.07 g (97%) of the title compound are obtained as a yellowish powder.

IR (KBr) v _max : 3700-3100 (OH), 1 770 (C = O of β-lactam), 1 695 (C = O of the PNB ester), 1 630 (pyridinium), 1 590 (phenyl), 1 518

(NO ₂ ), 1348 (NO ₂ ), 885 (NO ₂ ) cm- "H-NMR (DMSO-d ₆ ) δ: 1.14 (d, J = 6.1 Hz, 3H, CH ₃ CHO ), 1.33 (d, J = 6.3 Hz, 3H, CH ₃ CHS), 4.6-5.0 (m, CH ₂ N ⁺ ), 5.14 (d, J = 5.2 Hz , 1H, OH), 5.37 (center of ABq, J = 12.4 Hz, 2H, CH ₂ of PNB), 6.6-7.5 (m, 10H, phenyl of the phosphate), 7.69 ( d, J = 8.7 Hz, 2H, Ho of PNB), 8.0-8.4 (m, 4H, Hm of PNB, Hm of pyridinium), 8.4-8.8 (m, 1H, Hp of pyridinium), 9.08

(d, J = 5.6 Hz, 2H, Ho of pyridinium) UV (H ₂ O) X _max : 263 (ε 13325), 308 (ε 8915) Analysis: for C ₃₆ H ₃₆ N ₃ O ₁₀ SP-H ₂ O calculated: C 57.52% H 5.10% N 5.59% S 4.27% found: 57.76 4.96 5.36 4.35.

C. (5R, 6S) -3- [1- (R and

OH

Pd / C, H ₂

1 fa- S ^^ \ ^W / THF, ether, '

^ - H ^ f 0 buffer

0 ν CPhO) ^ PO "

COOPNB ^ rnu; ₂ ru

COO

To a solution of

"Boxylate diphenyl phosphate (0.60 g, 0.82 mmol) in humid tetrahydrofuran (33 mL) is added ether (33 mL), potassium phosphate monobasic sodium hydroxide buffer (17 mL, 0.15 M, pH 7.22) and 10%. iges palladium on carbon (0.60 g) and hydrogenates the resulting mixture for 1 h at 23 ⁰ C and a pressure of 2,76 bar (40 psi). Separate the two layers and extract the organic layer with water (3x7 ml). The aqueous layers are combined, filtered through a celite pad, washed with ether (3 × 20 ml) and chromatographies on a prep. Bondapak C-18 column (2.5 χ 9.5 cm) using water as the eluting solvent. 0.18 g (63%) of a mixture of diastereoisomers are obtained. The two diastereoisomers are separated by HPLC (Prep Bondapak C-18) using water as eluent. This gives 0.068 g (23%) of the isomer with the lower retention time, referred to as compound "B".

IR (KBr) v _max : 1770 (C = O of β-lactam), 1633 (pyridinium), 1 593 (carboxylate) cm "¹H-NMR (D ₂ O) δ: 1.20 (d, J = 6.3 Hz, 3H, _CH3 CHO), 1.42 (d, J = 6.9 Hz, 3H, CH ₃ CHS), 2.3-3.2 (m, 3H, H-4, H 6), 3.5-3.9 (m, 1H, SCH), 3.9-4.2 (m, 2H, H-5, _CH3 CHO), 4.3 to 5.1 (m, CH N ₂ "), 7.8-8.2 (m, 2H, Hm of pyridinium), 8.4-8.7 (m, 1H, Hp of

Pyridinium), 8.7-9.0 (m, 2H, Ho of pyridinium) UV (H ₂ O) X _max : 260 (ε 6727), 300 (ε 8245) [α $ = -39.3 ° (c , H ₂ O)

τ i / z ⁼ 12.6 h (measured at a concentration of 10 "* M in phosphate buffer pH 7.4 at 36.8 ⁰ C). As an isomer is also obtained with higher retention time, 0.081 g (28%), with compound "A" denotes. IR (KBr) v _max : 1755 (C = O of β-lactam), 1630 (pyridinium), 1590 (carboxylate) cm "¹H-NMR (D ₂ O) δ: 1.18 (d, J = 6 , 3 Hz, 3 H, CH ₃ CHO), 1.40 (d, J = 7.0 Hz, 3H, CH ₃ CHS), 2.84 (d, J = 9.3 Hz, 2H, H-4 ), 3.26 (dd, J = 2.7 Hz, J = 5.9 Hz, 1H, H-6), 3.4-4.2 (m, 3H, SCH, CH ₃ CH) O, H -5), 4.2-5.1 (m, CH ₂ N ⁺ ), 7.7-8.1 (m, 2H, Hm of;

Pyridinium), 8.3-8.65 (m, IH, Hp of pyridinium), 8.65-8.9 (m, 2H, Ho of pyridinium) UV (H ₂ O) X _max : 259 (ε 5694) , 296 (ε 6936) [a] f = + 96.9 ° (ο 0.56, H ₂ O)

15.6 h (measured at a concentration of 10 '"M in phosphate buffer pH 7.4 at 36.8 ⁰ C)

Example Production of

A. dl.-1- {2-mercapto-1-cyclohexyl) pyridinium methanesulfonate

MsOH

MsO

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 for 10 min under a nitrogen atmosphere treated with dl-cyclohexene sulfide [1.377 g (85 % purity), 0.0102 mol) and stirred for 25 h at 72 ° C. Excess pyridine is removed in vacuo. Traces are distilled together with water. The residue is mixed with water and chromatographies over a prep. Bondapak C-18 column (5χ 13 cm) with 0-2% acetonitrile in water as eluent. After lyophilization, 1.57 g (53%) of a colorless syrup are obtained.

v _max : 2500 (SH), 1625 (pyridinium), 1190 (SO ₃ -)

(DMSO-d _e ) S: 1.2-2.5 (m, 8H, cyclohexyl H), 2.32 (s, 3H, CH ₃ SO ₃ -), 2.82 (d, J = 9.8 Hz, SH), 3.0-3.5 (m, 1H, CHSH), 4.2-4.9 (m, 1H.CHN "), 8.0-8.3 (m, 2H, Hm of Pyridinium), 8.4-8.8 (m, 1H, Hp of pyridinium), 8.9-9.3 (m, 2H, Hp of pyridinium)

X _max : 214 (ε 5365), 258 (ε 3500)

for C ₁₂ H ₁₃ NO ₃ S ₂ -H ₂ O.

H 6.88% N 4.56%

6,46 4,65

B. (5R, 6S) -p-nitrobenzy! -3- [2- [iR or S) - {1-pyridinium)] - "! - (R or S) -cyclohexylthio] -6-t1- {R) -hydroxyetriyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate diphenylphosphate

IR (film) ¹ H-NMR

UV (H ₂ O) analysis: calculated: C 46.88% found: 46.61

O P

2) ClP (OPh),

4) NEt (iPr)

(PhO) ₂ PO '

To a cold (0 ⁰ C) solution of (SR.SSJ-p-nitrobenzyl-e-ti-IRj-hydroxyethylj-SJ-dioxo-i ^ azabicycloß .OL-heptaene ^ -carboxylate (1.37 g, 3 93 mmol) in 15 ml of acetonitrile (kept under a nitrogen atmosphere) are added diisopropylethylamine (0.822 ml, 4.7 mmol) and diphenylchlorophosphate (0.979 ml, 4.7 mmol) .The resulting solution is stirred for 30 min, treated with a solution of dl-1- (2-Mercapto-1-cyclohexyl) -pyridinium methanesulfonate (1.64 g, 5.66 mmol) in 4.7 mL of acetonitrile followed by diisopropylethylamine (0.822 mL, 4.7 mmol) the mixture is stirred 1 h at 0 ° C, Bondapak diluted with cold ^{(0 °} C) water (75 ml) and chromatographed using a prep. C-18 column eluting using 25-50% acetonitrile in water as eluent. lyophilization the appropriate fractions are obtained 1.9 g (53%) of the title compound: IR (KBr) v _max : 3700-3000 (OH), 1770 (C = O of β-lactam), 1,700 (C = O of PNB Ester), 1 628 (pyridinium), 1 590 (phenyl), 1 515

(NO _2), 880 _(NO2) cm ^{"1 1} H-NMR (D ₂ O) 5: 1.13 (d, J = 6.1 Η7, 3Η, _CH3 CHO), 1.2-2, 5 (m, 8H, cyclohexyl H), 2.7-3.5 (m, 4H, H-4, CHS), 3.5-4.4 (m, 2H, _CH3 CHO, H-5), 4.4-5.0 (m, 1H, CHN '), 5.30 (center of ABq, J = 12.8 Hz, _CH2 of PNB), 6.7-7.4 (m, 1 OH, phenyl ), 7.65 (d, J = 8.6 Hz, 2H, Ho of PNB), 7.9-8.4 (m, 4H, Hm of PNB, Hm of pyridinium), 8.4-8.8 (m, 1H, Hp of pyridinium), 9.0-9.4

(m, 2H, Ho of pyridinium) UV (H ₂ O) X _max : 263 (ε 9038), 309 (ε 6394) Analysis: for C ₃₃ H ₄₀ N ₃ O ₁₀ SP · H ₂ O calculated 59.16% H 5.35% N 5.31% found: 58.95 5.15 5.57

C. (5R, 6S) -3- [2 - [(Roder S) - (1-pyridinium)] - 1- {R or S) -cyclbhexylthio] -6- {1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate

Pd / C, H ₂

THF, ether, buffer

(PhO) ₀ PO '

To a solution of (5R, ois) - p -nitrodecylo-j- [z -nn-o-a-o-p-pu-nuuuuin-i-u-n-uuci S) -cyclohexylthio] -6- [1- (R) -hydroxyethyl] - Add 7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate diphenyl phosphate (1.85 g, 2.34 mmol) in wet tetrahydrofuran (96 mL) to ether (96 mL), potassium phosphate monobasic sodium hydroxide buffer (0.15 M, pH 7.22, 50 ml) and 10% palladium on carbon (1.9 g) and hydrogenates the resulting mixture for 1.25 h at 23 ° C under a pressure of 2.76 bar (40 psi) The organic layer is separated, washed with water (3 × 20 ml), the aqueous solutions are filtered through a celite pad, washed with ether (2 × 60 ml), and traces of organic solvents are removed by pumping and chromatographing on a prep.Bondapak C-18 column (4.5 x 9 cm) with 0-5% acetonitrile in water as the eluent to give, after lyophilization, 0.705 g (76%) of a mixture of diastereoisomers this diastereoisomer is by HPLC (prep.Bondapak C-18) with 4% aceto nitrile in water as eluent. The lower retention time diastereoisomer is designated compound "A" (0.29 g, 31%).

IR (KBr) v _m ": 1750 (C = O from β-lactam), 1620, (Sch, pyridinium), 1685 (carboxy-nate) cm"'H-NMR (D ₂ O) δ: 1.21 (d , J = 6.3 Hz, 3H, CH ₃ CHO), 1.4-2.5 (m, 8H, cyclohexyl H), 2.5-3.05 (m, 2H, H-4), 3, 05 to 3.25 (m, 1H, H-6), '3.3-3.7 (m, 1H, CHS), 3.9-4.3 (m, 2H, H-5, _CH3 CHO ), 4.3-4.8 (m, CHN ⁺ ), 7.8-8.2 {m, 2H, Hm of pyridinium), 8.3-8.7 (m,

1H, Hp of pyridinium), 8.8-9.1 (m, 2H, Ho of pyridinium) UV (H ₂ O) X _majt : 260 (ε 7123), 300 (ε 8685) [<*] § = + 6.2 ° (c 0.63, H ₂ O

τ _1/2 = 16.6 h (measured at a concentration of 10 ^"4 M in phosphate buffer pH 7.4 at 36.8 ⁰ C)

Analysis: ^ rC ₂₀ H ₂₄ N ₂ O ₄ S-SH ₂ O.

calculated 56.59% H 6.65% N 6.60% S7.55% Found: 56.83, 6.47, 6.59, 7.43.

The higher retention time diastereoisomer is referred to as compound "B" (0.35 g, 38%) IR (KBr) v _max : 1750 (C = O of β-lactam), 1 622 (Sch, pyridinium), 1588 (carboxylate) cm ^{-1 1} H-NMR (D ₂ O) δ: 1.19 (d, J = 6.4 Hz, 3H, _CH3 CHO), 1.3-2.5 (m, 8H, cyclohexyl H), 2.5-3.1 (m, 2H, H-4), 3.1-3.3 (m, 1H, H-6), 3.3-3.8 (m, 2H / H -5, CHS), 4.1 (center of m, 1H, _CH3 CHO), 4.25 to 4.7 (m, 1 H CHN ⁺ _1), 7.8-8.1 (m, 2H, Hm of pyridinium), 8.3-8.7

(m, 1Ή, Hp of pyridinium), 8.75-9.0 (m, 2H, Ho of pyridinium) UV (H ₂ O) X _max : 259 (ε.5992), 296 (ε 7646).

[α] ² ₀ ³ = + 65.3 ° (c 0.43, H ₂ O) '

τ _1/2 = 20.2 h (measured at a concentration of 10 ~ ⁴ M in phosphate buffer pH 7.4 at 36.8 ° C)

Example 15

A. (5R) -AMyi-3 - [(2-pyridinioethyl) -thio] - (6S) - [(1R) -hydroxyethyl] -7-oxo-1-azabicyclo {3.2.03hept-2-en-2 carboxylate diphenylphosphate

'. 1) ClPO (Οφ)

2

To a solution of (5R) -allyi-3,7-dioxo (6S) - [(1R) -hydroxyethyl] -1-azabicyclo [3.2.0] heptane (2R) -carboxylate (473 mg, 1.87 mmol) in 6 mL of (CH ₃ CN are added at about -1O ⁰ C under a nitrogen atmosphere diisopropylethylamine (0.42 mL, 2.4 mmol) and then Diphenytchlorphosphat (0.50 mL, 2.4 mmol). The mixture is stirred 30 min at 0 ⁰ C, cooled to -15 ° C, an oily suspension of N- (2-mercaptoethyl) pyridinium chloride (527 mg, 3.00 mmol) in a 1 ml CH ₃ CN at a content of 5 drops of DMF and then diisopropylethylamine (0.42 mL, 2.4 mmol) The mixture is stirred for 30 min at -15 ° C, diluted with 20 mL water, this mixture is purified directly on a reverse phase silica gel column (C _ia PrepPAK, 12 g , Waters Associates) eluting with water (200 ml) 10% CH ₃ CNVH ₂ O (100 ml), 20% CH ₃ CN / H ₂ O (100 ml), 30% CH ₃ CN / H ₂ O (100 ml and then with 40% CH ₃ CN / H ₂ O (100 mL). Collect the appropriate fractions, remove the organic solvent with a Vacuum pump and lyophilized to give 786 mg (1.26 mmol, yield 67.3%) of Titalverbindung as a brownish powder.

'H-NMR (DMSO-d ₆ , CFT-20) δ: 1.16 (3H, d, J = 6Hz, 1 ^ CH ₃ ), 2.6-3.7 (m), 3.75- 4.3 (2H, m, 5-H and 1'-H), 4.65 (2H, m, -CO ₂ CH ₂ -), 4.87 (2H, t, J = 6Hz, -CH ₂ -N ⁺ ), 5-6.2 (3H, m, olefinic protons), 6.6-7.4 (m, aromat. Protons), 8.15 (2H, t, J = 7 Hz, aromat. Protons nitrogen to nitrogen), 8.63 (1H, t, J = 7 Hz, aromat.Proton para to nitrogen), 9.07 ppm (2H, d, J = 7 Hz, aromat.protons ortho to nitrogen) IR (film ) v: 3400 (OH), 1 770 (β-lactam), 1 690 (ester), 1 625 (pyridinio).

B. (5R) -3- [(2-pyridinioethyl) -thioH6S) - [C Rj-

CO ₂ K

To a solution of

(5R) allyl 3 - [(2-Pyridinioethyl) thio] - {6S) - [(1R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2- carboxylate diphenyl phosphate (156 mg, 0.25 mmol) in 2 ml CH ₃ CN are added successively at about 22 ⁰ C, a solution of Kaiium-2-ethylhexanoate in EtOAc (0.5 M, 0.6 ml, 0.3 mmol), triphenylphosphine (15 mg, 0.057 mmol) and tetrakistriphenylphosphine-palladium (15 mg, 0.013 mmol) and the mixture is stirred for 2 h at about 22 ⁰ C under a nitrogen atmosphere. After adding 7 ml of anhydrous Et ₂ O, the precipitate is filtered, washed with 7 ml of anhydrous Et ₂ O and dried in vacuo to give 101 mg of a brownish solid. This is purified by reverse phase column chromatography (C ₁₈ PrepPAK, 12 g. Waters Associates) eluting with water. Suitable fractions (Fr.7-12, 20 ml each) are collected and lyophilized to give 53 mg (0.16 mmol, yield 64%) of the title compound as a yellowish powder. This material is contaminated with potassium diphenyl phosphate and potassium 2-ethylhexanoate

¹ H-NMR (D ₂ O, CFT-20) δ: 0.80 (t, J = 6.4 Hz, Me of ethylhexanoate), 1.21 (3H, d, J = 6.3 Hz, 1'-Me ), 2.93 (2H, dd, J, _ ₅ = 9 Hz, J _gem = 4 Hz, 1 -HS), 3.28 (1H, dd, J ₈ _, = 6.2 Hz, J ^ ₅ = 2.5 Hz, 6-H), 3.42 (2H, t J = 6 Hz, -CH ₂ S), 3.98 (1H, td, J ₅ _, = 9 Hz, J ₅ ^ = 2, 5 Hz, 5-H), 4.15 (1H, q, J = 6.2 Hz, 1'-H), 4.80 (2H, t, J = 6.0 Hz, -CH ₂ N *) , 7-7.5 (m, phenyl protons of diphenyl phosphate), 8.03 (2H, m, Hm of pyridinium), 8.56 (1H, m, Hp of pyridinium) and 8.81 ppm (2H, d, J = 6.5 Hz, Ho of pyridinium).

Example 16 Production of

3- {2- (N-methyl-thiomorpholinium) -ethyl-thio] -6a- [1 '- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene 2-carboxylate

A. N-Methyl-N- (2-mercaptoethyl) thiomorpholinium methanesulfonate

MeN

I) MsOH

²⁾ Δ

MsO

Pre-cooled (ice-bath) N-methylthiomorpholine (5.00 g, 42.7 mmol) (see JM Lehn and J. Wagner, Tetrahedron, 26, 4227 [1970]) is added methanesulfonic acid (1.47 ml, 20.5 mmol) ) and ethylene sulfide (1.30 ml, 21.4 mmol), the mixture is heated to 65 ° C for 24 h and diluted with 25 ml of water. The aqueous solution is washed with diethyl ether (3 x 25 ml), pumped under vacuum and poured onto a

Silica gel reverse phase column. The title compound is eluted with water. The appropriate fractions are combined and concentrated to give the thiol as an oil (4.80 g, 86% yield), IR (FI) _m : 2550 cm- ¹ (schw, SH),

¹ H-NMR (DMSO-d _e ) δ: 3.25-2.95 (6H, m, CH ₂ N ⁺ ), 3.32 (3H, s, CH ₃ N ^T ), 3.20-2, 65 (7H, m, CH ₂ S, SH) and 2.32 ppm (3H, s, CH ₃ SO ₃ ). B. p-nitrobenzyl S - ^ - IN-methyl-thiomorpholine-diphenyl phosphate J-ethylthioj-ea-ii'-tRJ-hydroxyethyl-y-oxo-i-azabicyclo [3.2.0] hept-2-ene-2-carboxylate

D EtN

2). CLPO (Οφ)

0 "

CO ₂ PNB

  ca

CO PNB OP (CX?).

0 · _

A cold (ice bath) solution of p-nitrobenzyl 6α- [1 '- (R) -hydroxyethyl] -3,7-dioxo-1-azabicyclo [3.2.0] heptane-2-carboxylate (557 mg, 1.60 mmol) in 8 ml of CH ₃ CN is treated dropwise with diisopropylethylamine (0.336 ml, 1.92 mmol) and diphenylchlorophosphate (0.400 ml, 1.92 mmol), stirred for 30 min, treated with the reaction mixture

N-methyl-N- (2-mercaptoethyl) thiorhorpholinium methanesulfonate (893 mg, 2.29 mmol) in 4 mL of CH ₃ CN and with diisopropylethylamine (0.336 mL, 1.92 mmol) and stirred for 30 min. The solution is diluted with 20 ml of water and poured onto a silica gel reverse phase column. The desired compound is eluted with a 50% acetonitrile-water mixture, the appropriate fractions combined, pumped under vacuum for 2 h and lyophilized to give the title compound (1.01 g, 85%). IR (nujol) v _max : i 760 (s, β-lactam C = O), 1510 cm ^-1 (st, NO ₂ ) cm ^-1

'H-NMR (DMS0-d ₆₎ δ: 8.25 (2H, d, J = 8.8 Hz, H-aromat.), 7.70 (2H, d, J = 8.8 Hz, H aromatic), 7.33-6.84 (1OH, m, H-arom.), 5.37 (2H, center of ABq, J = 14.2Hz, CH ₂ ), 5.14 (1H, d , J = 4.5 Hz, OH), 4.35-3.80 (2H, m, HT and H-5), 3.75-3.45 (6H, m, CH ₂ N ⁺ ), 3, 31 (3H, s, CH ₃ N ^+), 3.45 to 2.75 (9H, m, CH ₂ S, H-6 and H-4) and 1.15 ppm (3H, d, J = 6, 2 Hz, CH ₃ ). C. 342- {N-Wiethylthiorio- horphinium) ethylthio] -6a- [1 '- {R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate

  OH

A solution of '.' '.

p-nitrobenzyl-3- [2- (N-methyl-thiomorpholinium diphenyl phosphate) ethylthio] -6a- [1 '- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2 -en-2-carboxylate (1.31 g, 1.76 mmol) in 0.1 M pH 7.4 phosphate buffer (48.8 mL), tetrahydrofuran (20 mL) and diethyl ether (20 mL) is hydrogenated at 2 for 1 h , 76 bar (40 psi) over 10% Pd / C (1.5 g) in a Parr shaker. The reaction mixture is diluted with 40 ml of diethyl ether and the phases are separated. The organic phase is washed with water (2 × 5 ml), the aqueous phases are combined, filtered through a No. 52 hardened filter paper, washed with diethyl ether (2 × 20 ml) and vacuum is applied. The aqueous solution is poured on

a silica gel reverse phase column and elutes the desired carbapenem with 5% acetonitrile-water. The appropriate fractions are combined and lyophilized to give the title compound as an amorphous solid (205 mg, 31 ⁰ A).

IR (Nujol) v _max : 1 750 (st, β-lactam C = O), 1 590 cm -1 (st, C = O)

¹ H-NMR (D ₂ O! Δ: 4.25-3.95 (2H, m, HT, H-5), 3.70-3.40 (6H, m, CH ₂ N "), 3, 55 (1H, dd, J = 6.1 Hz, J = 2.6 Hz, H-6), 3.08 (3H, s,

CH ₃ N ⁺ ), 3.25-2.75 (8H ₁ CH ₂ S, H-4) and 1.24 ppm (3H, d, J = 6.4 Hz, CH ₃ ) UV (H ₂ O; c 0.062) X _max : 299 (ε 10962)

r, _{/ 2} = 17.7 h (0.1 M pH 7 phosphate buffer, 37 "C).

Example 17

Production of (5R, A. 1-Methyl-1- {2-mercaptoethyl) morpholinium trifluoro-methanesulfonate

CF ₃ SO ₃ H

me

CF ₃ SO ₃ ^

To N-methylmorpholine (3.29 ml, 0.030 mol) is added dropwise at 10 ⁰ C dropwise trifluoromethanesulfonic acid (1.327 ml, 0.015 mol) and then ethylene sulfide (0.89 ml, 0.015 mol), the resulting yellow-brown solution heated for 18 h N ₂ on the oil bath at 50 to 60 ⁰ C, then removed volatile material in vacuo and the remaining oil in 10 ml of water. The aqueous solution is washed with diethyl ether (3x5 ml), then removed residual organic solvent in vacuo and gives the resulting aqueous solution to a reverse phase column C _ia, the one with water, then 5% acetonitrile-H ₂ 0 and finally 10% Acetonitrile-H ₂ 0 eluted. Evaporation of the relevant fractions yields a white solid which is dried in vacuo (P ₂ O ₅ ) to give the product (1.92 g, 41%).

IR (KBr) 7 ^ 2560 (-SH) CiTi- ¹

¹ H-NMR (d _r acetone) δ: 4.25-3.6 (m, 8H), 3.49 (s, 3H, N-Me), 3.35-2.7 (m, 5H).

B. p-Nitrobenzoyl (5R, 6S) -3- [2- (1-methylmoholino) ethylthio] -6 - [{R) -1-hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate diphenylphosphate

CO ₂ ? N3

To a solution of p-nitrobenzyl- (5R, 6S) -6 - [(R) -1-hydroxyethyl] -3,7-dioxo-1-azabicyclo [3.2.0] heptane-2-carboxylate (0.348 g, 1 , 0 mmol) in 25 mL of dry acetonitrile are added dropwise diisopropylethylamine (0.191 mL, 1.1 mmol) followed by diphenyl chlorophosphate (0.228 ml, 1.1 mmol) at ⁰ ° C under N _2, is to Istündigem stirring at ⁰ ° C Diisopropylethylamine (0.226 mL, 1.3 mmol) to the resulting enol phosphate followed by 1-methyl-1- (2-mercaptoethyl) -morpholinium trifluoromethane-sulfonate (0.373 g, 1.2 mmol). The reaction mixture is stirred for 1.5 h at room temperature, concentrated in vacuo, the remaining material is taken up in water and added to a C ₁₈ reverse phase column. Elution with water, then 20% acetonitrile-H ₂ O and finally 30% acetonitrile-H ₂ O and subsequent lyophilization of the relevant fractions gives the product (0.360 g, 40%) as an amorphous solid.

IR (film): 3300 (-OH), 1,770 (β-lactam CO), 1,700 (-CO ₂ PNB) cm "'

'H-NMR (d ₆ -acetone) δ: 8.25, 7.80 (Abq, J = 8.6 Hz, 4H, arom.), 7.4-6.8 (m, 10H, diphenyl phosphate), 5.56, 5.27 (ABq, J = 14.2Hz, 2H, benzyl.), 4.42 (d of t, J = 9.2Hz, J '= 2.7Hz, 1H, H- 5), 4.1-2.7 (m, 17H), 3.40 (s, 3H, N-Me), 1.22 (d, J = 6.2 Hz, 3H, -CHMe). , , ·.

C0 "? ²

To a solution of

p-Nitrobenzyi (5R, 6S) -3- [2- (1-methylmorpholino) ethylthio] -6- [(R) -I-h

Diphenyl phosphate (0.360 g, 0.49 mmol) in 13 mL of phosphate buffer (0.05 Μ pH 7.4) is added 0.36 g of 10% palladium-on-carbon, 20 mL

Tetrahydrofuran and 20 ml of diethyl ether and hydrogenated de this mixture (Parr) for 1 h under a pressure of 2.2 bar (32 psi). The

The mixture is filtered through Celite, the Fiiterkissen washed with water and diethyl ether, the aqueous phase is separated off and the pH is adjusted to 7.0 with additional pH 7.4 phosphate buffer. After removal of residual organic solvents in vacuo, the aqueous solution is added to a C ₁₈ reverse phase column. Elution with water and lyophilization of the relevant fractions yields 0.130 g of an amorphous solid. This material is purified again by reverse phase HPLC to give the pure product (0.058 g, 34%) as an amorphous solid.

IR (KBr) v _max : 3420 (br, OH), 1 7B0 (β-lactam CO), 1590 (-CO ₂ -) cm ' ¹ "

'H-NMR (D ₂ O) δ: 4.35-2.77 (m, 17H), 3.18 (s, 3H, N-Me), 1.23 (d, J = 6.3 Hz, 3H, CHMe)

UV (H ₂ O) X _max : 300 (ε 6344) nm

τι / 2 (pH 7.4.36.8 ° C) = 18.5 h. ,

Claims (5)

Invention claim: 1. A process for preparing a quaternary Aminthiolverbindung the general formula VIl HS-AR "X" ^ _n * ο Vl _R 1Ü _R 1 ^ wherein A is cyclopentylene, cyclohexylene or II -C c - R ¹¹ R ¹³ in which R ¹⁰ , R ", R ' ² and R' ³ independently of one another are a hydrogen atom or a C 1-4 -alkyl group, X ^{e is} a counteranion and R ' ^{4 is} a substituted or unsubstituted, mono-, bi- or polycyclic, is an aromatic or non-aromatic heterocyclic radical containing at least one nitrogen atom in the ring and attached to A via a ring nitrogen atom, forming a quaternary ammonium group, characterized in that a sulfide of the general formulas 10 SR ¹² or C-C _R 11 _R 13 wherein R ¹⁰ , R ' ¹ , R' ² and R ¹³ each independently represent a hydrogen atom or a C 1-4 alkyl group, with a strong acid and either (a) with a heteroaromatic amine of the formula this remainder a substituted or unsubstituted, mono-, bi- or polycyclic, aromatic, heterocyclic radical containing at least one nitrogen atom in the ring, which can be quaternized and which is bonded to a carbon atom of the substituent A, or
(b) with a heterocyclic amine of the formula α f. wherein the rest is a substituted or unsubstituted, mono-, bi- or polycyclic, non-aromatic, heterocyclic radical containing at least one nitrogen atom in the ring, which can be quaternized and is bonded by the substituent R ' ⁶ and to a carbon atom of the substituent A, and wherein R ¹⁸ either (i) an optionally substituted, aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, heteroaraliphatic, heterocyclyl-aliphatic or araliphatic radical or an aryl, heteroaryl or heterocyclyl radical (ii) a divalent phenylene or C ₁ _ ₄ -Alkylengrupe means which with the ring of the general formula is connected so that a bridged, polycyclic group is formed, implements. 2. The method according to item 1, characterized in that one carries out the reaction at a temperature of about -2O ⁰ C to about 100 ⁰ C. 3. The method according to item 1 or 2, characterized in that is used as the strong acid hydrochloric acid, hydrobromic acid, methanesulfonic acid, p-toluenesulfonic acid or trifluoromethanesulfonic acid. 4. The method according to item 1, characterized in that one carries out the reaction in the presence of a non-polar 3, organic solvent, preferably methylene chloride, benzene, xylene or toluene. 5. The method according to any one of items 1 to 4, characterized in that one carries out the reaction without additional solvent, if the amine and sulfide reagents are liquid or if the amine reagent is a solid, the sulfide reagent is liquid and the solid amine in the liquid sulfide reagent is soluble.