CS247195B2 - Process for producing carbapenem derivatives - Google Patents

Abstract

A process for the production of carbapenem derivatives of the general formula I, where the general symbols have the meanings given below, and their pharmaceutically acceptable salts, characterized in that an intermediate of the general formula II, where the general symbols have the meanings given below, is reacted with an alkylating agent of the general formula R5 - X', where the general symbols have the meanings given below, to form a compound of the general formula I', where the general symbols have the meanings given below, and optionally replacing the anion X' with another anion and optionally removing the carboxyl-protecting group R2 and optionally converting the compounds of the formula I into pharmaceutically acceptable salts. The compounds of the formula I are useful for the treatment of bacterial infections.

Description

A process for the production of carbapenem derivatives of general formula I, where the general symbols have the meanings given below, and their pharmaceutically acceptable salts, characterized in that an intermediate of general formula II, where the general symbols have the meanings given below, is reacted with an alkylating agent of general formula

R ⁵ - X', where the general symbols have the meanings given below, to form a compound of general formula I', where the general symbols have the meanings given below, and optionally replacing the anion X' with another anion and optionally removing the carboxyl protecting group R2 and optionally converting the compounds of formula I into pharmaceutically acceptable salts.

Compounds of formula I are useful for treating bacterial infections.

(II) (B2) (51) Int. Cl. ⁴ C 07 D 205/08, C 07 D 487/04

The present invention relates to novel carbapenem antibiotics which have a substituent of the general formula at position 2

-S-A· 'W R

NR ⁶ where

A is an alkylene with 1 to 6 carbon atoms with a straight or branched chain, ς

R is an optionally substituted aliphatic, cycloaliphatic, cycloaliphatic-co-aliphatic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, heterocyclic or heterocycloaliphatic radical and the ring of the formula represents a nitrogen-containing aromatic heterocycle attached to the alkylene group A by a ring carbon atom and quaternized by the substituent R.

A number of beta-lactam derivatives containing a carbapenem core are described in the literature.

These carbapenem derivatives are useful as antibacterial agents and/or beta-lactamase inhibitors.

The original carbapenem compounds were natural products, such as thienamycin with the formula

SCH ₂ CH ₂ NH ₂

COOH and were prepared by fermentation of Streptomyces cattleya (U.S. Patent No. 3,950,357). Thienamycin is an exceptionally potent broad-spectrum antibiotic that is particularly effective against various Pseudomonas species, organisms that are known to be resistant to beta-lactam antibiotics.

Other natural compounds containing a carbapenem core include olivanic acid derivatives, such as the antibiotic MM 13902 of formula ch ₃

-sch=chnhcoch ₃

COOH Claimed in US Patent No. 4,113,856, Antibiotic MM 17880 Formula

CH, claimed in US patent 4 claimed in US patent no

HO ₃ S0 sch ₂ ch ₂ nhcoch ₃

COOH

162,304, antibiotic MM 4550A formula

HOjSO

CH, and ,s-ch = chnhcoch ₃ ⁰ COOH

172 129, and antibiotic 890Ag formula

CH,

NO

II ^sch=chnhcch ₃

COOH claimed in US Patent No. 4,264,735.

In addition to natural compounds, the compound desacetyl 890Ag of the formula

O COOH is disclosed in U.S. Patent No. 4,264,734 and is prepared by enzymatic deacylation of the corresponding N-acetyl compound. Various derivatives of natural olivic acids have also been synthesized, for example compounds of the formula ch ₃ n ₂ oo

^s<0)n w ^NHC0CH3

CO ₂ RJ where CO^R^ is a free, esterified carboxyl group or a carboxyl group in salt form n is 0 or 1 and R ₂ is a hydrogen atom, an acyl group or a group of the formula where R^ is a salt-forming ion or methyl or ethyl. These compounds are disclosed in European Patent Application 8,885.

U.S. Patent No. 4,235,922 (see also European Patent Application 2,058) claims a carbapenem derivative of the formula

sch ₂ ch ₂ nh

COOH while British Patent Application No. 1,598,062 claims the isolation of the compound

SCH ₂ CH ₂ NHCOCH ₃

COOH

from Streptomyces fermentation medium.

Carbapenems that are unsubstituted at position 6 have also been synthesized. Thus, in U.S. Patent No. 4,210,661, compounds of the formula s -r ₂ are disclosed.

COOH where R ₂ is phenyl or substituted phenyl, U.S. Patent No. 4,267,177 discloses compounds of the formula

s-n,

COOH where is an optionally substituted pyridyl group and U.S. Patent No. 4,255,441 discloses compounds of the formula

S—CR ₂ ⁼ CR^Rq

0=

COOH where _R2 and Rg are hydrogen or alkyl and R^ is substituted phenyl and is 1 or 2 and U.S. Patent No. 4,282,236 discloses compounds of the formula,CH _{= CR1R2}

COOH where R is hydrogen and R ₂ is cyano or CO ₂ Rg where Rg is hydrogen, alkyl, aryl or arylalkyl.

Carbapenems of general formula

R^NH—t

COOH

8 where R is hydrogen or acyl and R is hydrogen or substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, aryl, arylalkyl, arylalkenyl, arylalkynyl, heteroaryl, heteroarylalkyl, heterocyclyl or heterocyclylalkyl are disclosed in U.S. Patent No. 4,218,463. No claims are made to any heteroarylalkyl or

substituents R of the type where A is alkylene and the n?-r ⁵ part of the molecule is a quaternary nitrogen containing aromatic heterocycle attached to the alkylene group A through a carbon atom in the cycle.

The natural product thienamycin has the absolute configuration 5R, 6S, 8R. This isomer, as well as the other seven thienamycin isomers, can be obtained by total synthesis as disclosed in U.S. Patent No. 4,234,595.

Total synthesis procedures are also disclosed, for example, in U.S. Patent Nos. 4,287,123,

269,772, 4,282,148, 4,273,709, 4,290,947 and in European Published Patent Application No. 7,973. The key intermediate in the claimed synthetic methods is

OH

O CO ₂ pNB where pNB is p-nitrobenzyl.

Due to the exceptional biological activity of thienamycin, a large number of derivatives have been prepared and documented in the literature. Of these compounds, the following are

1) N-formimidoylthienamycin formula

OH sch ₂ ch ₂ n=c-nh ₂

COOH documented in European Published Patent Application No. 6,639

2) N-heterocyclic derivatives of thienamycin of general formulas I and II

O

COOH (I) (II) where the bifunctional ring may contain additional unsaturation in the ring and where n is an integer selected from 1 to 6, p is 0, 1 or 2, is hydrogen, alkyl or aryl and Z is imino, oxo, hydrogen, amino or alkyl are documented in U.S. Patent No. 4,189,493,

3) substituted N-methylene derivatives of thienamycin of the general formula

OH sch ₂ ch ₂ n = cCOOH

2 where X and Y are hydrogen, R, OR, SR or NR R , where R is substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocyclyl or heterocyclylalkyl and R and R are hydrogen or R are documented in U.S. Patent No. 4,194,047,

4) compounds of the general formula ? ⁿ 3 sch ₂ ch ₂ nr ¹ r ²

COOH

12 where R is aryl, alkyl, acyl or arylalkyl and R and R are independently selected from hydrogen and acyl (including acyl of the type

O

-C - R where R ¹¹ can be, among others, alkyl substituted with a quaternary ammonium group, for example

About ©

-c-ch ₂ - i are listed in U.S. Patent No. 4,226,870

5) compounds of the formula

OR ³

About COOH

1 where R is a hydrogen atom, acyl or a monovalent optionally substituted hydrocarbon radical, R is optionally substituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkenylalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl or heteroarylalkyl and R is acyl (including acyl of the type

About me

-C - R where R is alkyl substituted with a quaternary ammonium group, e.g.

About us

-c-ch ₂ ), are disclosed in British Patent No. 1,604,276 (see also US Patent No. 4,235,917)

6) compounds of the general formula

OH ₂ NR ⁵ R ⁶ R ⁷

AND

6 7 where R , R and R are independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkenylalkyl, cycloalkylalkyl, aryl, in U.S. Patent No. 4,235,920,

7) compounds of the formula arylalkyl, heteroaryl or heteroarylalkyl are listed

OR ³ ήτίΧ o

R ⁵ t ® , ₂

SCH ₂ CH ₂ NC = NR ¹ R' ^!

1« cox

2 where R and R independently of the others are radicals of the type defined for R, a hydrogen atom or a nitro group, hydroxyl, alkoxy with 1 to 6 carbon atoms, amino group, alkylamino group with 1 to 6 carbon atoms, dialkylamino group or trialkylamino group with 1 to 6 carbon atoms 1 2 in each of the alkyls and the additional anion present in the latter case or R and R can be joined together to form, together with the nitrogen atom to which they are attached, a substituted or unsubstituted monocyclic or bicyclic heteroaryl or heterocyclic radical, containing 4 to 10 ring atoms, one or more of which may be additional heteroatoms selected from the group comprising oxygen, sulfur and nitrogen, R is cyano or substituted or unsubstituted carbamoyl, carboxyl, alkoxycarbonyl with 1 to 11 carbon atoms, alkyl with 1 to 10 carbon atoms, alkenyl with 2 to 10 carbon atoms carbon, alkynyl with 2 to 10 carbon atoms, cycloalkyl with 3 to 10 carbon atoms, cycloalkylalkyl with 4 to 12 carbon atoms, cycloalkylalkenyl with 5 to 12 carbon atoms, cycloalkenyl with 3 to 10 carbon atoms, cycloalkenylalkenyl with 5 to 12 carbon atoms, cycloalkenylalkyl with 4 to 12 carbon atoms, aryl with 6 to 10 carbon atoms, arylalkyl with 7 to 16 carbon atoms, arylalkenyl with 8 to 16 carbon atoms, arylalkynyl with 8 to 16 carbon atoms or monocyclic or bicyclic heteroaryl, heteroarylalkyl, heterocyclyl or heterocyclylalkyl with 4 to 10 ring atoms, one or more of which are heteroatoms selected from oxygen, sulfur and nitrogen and where the alkyl residue of the heteroarylalkyl or heterocyclylalkyl radical contain

2 from 1 to 6 carbon atoms, the substituent or substituents on R, R, R or on the ring formed by the connection of R and R are chlorine, bromine, iodine, fluorine, azido, alkyl with 1 to 4 carbon atoms, mercapto, sulfo, phosphono, cyanothio (-SCN), nitro, cyano, amino, hydrazino, substituted amino or hydrazino groups with up to three alkyls with 1 to 6 carbon atoms, hydroxyl, alkoxy with 1 to 6 carbon atoms, alkylthio with 1 to 6 carbon atoms, carboxyl, oxo, alkoxycarbonyl with 1 to 6 carbon atoms in alkoxy, acyloxy with 2 to 10 carbon atoms, carbamoyl, alkylcarbamoyl with 1 to 4 carbon atoms in alkyl or dialkylcarbamoyl with 1 to 4 carbon atoms in the alkyl, R ₇ is a hydrogen atom, acyl or a residue of the type defined for R , R is alkyl with 1 to 10 carbon atoms, substituted carbonylmethyl, alkoxyalkyl with 1 to 6 carbon atoms in both parts of the substituent, cycloalkoxyalkyl with 3 to 6 carbon atoms in the cycloalkoxy and with 1 to 6 carbon atoms in the alkyl, alkanoyloxyalkyl with 2 to 12 carbon atoms, partially or fully halogenated alkyl with 1 to 6 carbon atoms, where the halogen atoms are chlorine, bromine or fluorine, aminoalkyl, alkenyl or alkynyl with 2 to 10 carbon atoms, acyl, alkoxycarbonylalkyl with 3 to 14 carbon atoms, dialkylaminoacetoxyalkyl with 4 to 21 carbon atoms, alkanoylaminoalkyl with 2 to 13 carbon atoms, arylalkyl with 1 to 3 carbon atoms in the alkyl and where the aryl radical contains from 6 to 10 carbon atoms, monocyclic or bicyclic heteroarylalkyl or heterocyclylalkyl containing 4 to 10 ring atoms, up to 3 carbon atoms in the alkyl radical and 1 to 4 heteroatoms selected from oxygen, sulfur and/or nitrogen atoms, arylalkyl or heteroarylalkyl substituted on the nucleus, where the substituent is a chlorine, fluorine, bromine or iodine atom or alkyl with 1 to 6 carbon atoms, aryl or aryl substituted on the nucleus containing 6 to 10 carbon atoms and where any substituent on the nucleus is hydroxyl, alkyl with 1 to 6 carbon atoms, chlorine, fluorine or bromine atom, arylalkoxyalkyl, alkylthioalkyl with 2 to 12 carbon atoms, cycloalkylthioalkyl with 4 to 12 carbon atoms, acylthioalkyl with 2 to 10 carbon atoms in acyl moieties and 1 to 6 carbon atoms in the alkyl or phenylalkenyl, where alkenyl has 2 to 6 carbon atoms, R is substituted or unsubstituted alkyl with 1 to 10 carbon atoms, alkenyl with 2 to 10 carbon atoms or alkynyl with 2 to 10 carbon atoms, ring-substituted or unsubstituted cycloalkyl, cycloalkenyl, cycloalkenylalkyl and cycloalkylalkyl with 3 to 6 carbon atoms and up to 6 carbon atoms in any chain, aryl with 6 to 10 carbon atoms, arylalkyl with 6 to 10 carbon atoms and 1 to 6 carbon atoms in the alkyl moiety, monocyclic or bicyclic heteroaryl or heteroarylalkyl containing 4 to 10 carbon atoms in the ring, one or more of which is an oxygen, nitrogen or sulfur atom and 1 to 6 carbon atoms in the alkyl moiety and the substituents in the ring or on the chain are chlorine, bromine, iodine, fluorine, azldo, cyano, amino, alkylamino with 1 to 6 carbon atoms, dialkylamino with 1 to 6 carbon atoms in each of the alkyls or trialkylamino with 1 to 6 carbon atoms in each of the alkyls and in the latter case an anion, hydroxyl, alkoxyl with 1 to 6 carbon atoms, alkylthioalkyl with 1 to 6 carbon atoms in each of the alkyls, carboxyl, oxo, alkoxycarbonyl with 1 to 6 carbon atoms in the alkoxyl, acyloxy with 2 to 10 carbon atoms, carbamoyl, alkylcarbamoyl with 1 to 4 carbon atoms, dialkylcarbamoyl with 1 to 4 carbon atoms in each of the alkyls, cyanothio (-SCN) or nitro, R® is hydrogen, hydroxyl, mercapto, R, -0R, -SR, or 12 12

-NR R , where R, R and R have the meaning given above, * 4 4 4

X is hydroxyl, mercapto, amino, acyloxy, -OR, -SR, -NHR,

-NR ⁴ , OM, OQ or if the compound is in the form of a double ion, -O , in which case A is absent,

And, if the compound is not in the form of a double ion, the ion is

M is a pharmaceutically acceptable cation and

Q is a protecting group as defined above, are claimed in British Patent No. 1,604,275 and

8) compounds of the general formula

OH sch ₂ ch ₂ w COO where

R ©li ( j attached to the nitrogen atom of the amino group of thienamycin represents a mono- or polycyclic nitrogen atom containing heterocyclic group and

R is hydrogen, substituted or unsubstituted alkyl, aryl, alkenyl, heterocyclylalkenyl, arylalkenyl, heterocyclylalkyl, arylalkyl, -NR ₂ , COOR, CONR ₂ ,

-0R or CN are claimed in European Published Patent Application No. 21,082.

Among the compounds disclosed in U.S. Patent No. 4,235,920 are compounds of the formula © fCj sch ₂ ch ₂ n(ch ₃ ) ₃ and

O COOH where A is a pharmaceutically acceptable anion.

The above-mentioned quaternary amine derivatives are also described in Recent Advances in the Chemistry of beta-Lactam Antibiotics, Royal Society of Chemistry, London, 1981, pp. 240-254, where the antibacterial activity is on average about 1/2 to 2/3 of that of thienamycin.

In addition to the above, carbapenem derivatives also have a wide range of substituents at position 6. For example:

1) European Published Application No. 40 408 claims compounds of the formula

O COOH where is a hydrogen atom, methyl or hydroxyl and is a monovalent organic group including but not limited to heterocyclylalkyl,

2) European Published Patent Application No. 8,514 claims compounds of the formula

COOH where is an optionally substituted pyrimidinyl group and R ₂ is a group CR-^R^j, where R ₃ is a hydrogen atom or hydroxyl, and Rj is a hydrogen atom, alkyl, benzyl or phenyl or a ky ring, is a hydrogen atom or the group R^ is a hydrogen atom or alkyl and Rq together form a carbocyclic

3) European Published Patent Application No. 38,869 discloses compounds of the formula

R ⁶

R ⁷

SR ⁸

O

COOH

7 A where R°, R° and R° are independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, alkenyl or alkynyl with 1 to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl with 3 to 6 carbon atoms in the cycloalkyl ring with 1 to 6 carbon atoms in the alkyl part, aryl such as phenyl, arylalkyl, arylalkenyl and arylalkynyl, where the aryl part of the molecule is phenyl and the aliphatic part has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, where the substituent or substituents related to the above-mentioned residues are selected from the group consisting of —X° halogen (chloro, bromo, fluoro), —OH hydroxyl, —OR*· alkoxyl, aryloxyl O

M 12

-OCNR R carbamoyloxy O

-<!!nr1r2 carbamoyl 1 2

-NR R amino group amidino group

-NO ₂ nitro group, © 1 1

-N(R ), a tri-substituted amino group (R is independently selected)

Γ ¹ 2 .

-C=NOR oximino group

-SR ⁴ alkyl- and arylthio group 1 2

-SOjNR R of the sulfonamido group O ¹¹ 1 2

-NHCNR R ureido group O

1 ¹¹ 2

-R CNR - amido group

-COjH carboxy group

-CO ₂ R ⁴ carboxylate O ¹¹ 1

-CR acyl O # 1

-OCR acyloxy group

-SH mercapto group O

-l§R ⁴ alkyl- and arylsulfinyl O

II 1

-SR alkyl- and arylsulfonyl II O

-CN cyano group

-Nj azido group where, given the above list of substituents on R®, R? and R® are substituents and R® are independently selected from the group consisting of hydrogen, alkyl, alkenyl and alkynyl with 1 to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl with 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl part of the molecule, aryl, such as phenyl, arylalkyl, arylalkenyl and arylalkynyl, where the aryl part is phenyl and the aliphatic part has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl and where the heteroatom or heteroatoms in the above-mentioned heterocyclic parts of the molecule are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and where the alkyl parts attached to these heterocyclic parts have 1 to 6 carbon atoms (see also European Patent Publication Nos. 1,627, 1,628, 10 317, 17,992, 37,080, 37,081 and 37,082).

4) European Published Patent Application No. 24,832 discloses as antibacterial agents compounds of the formula

7' ^ch 3 ^{ch s} ” ¹²

J-N--X.

o co ₂ h where R ¹ is a hydrogen atom or a group selected from substituents including OH, OSO^H or a salt or alkyl ester thereof with 1 to 4 carbon atoms, OR ² , SR ³ , OCOR ² , OCO ₉ R ³ or OCONHR ³ , where R ² 3 is alkyl with 1 to 6 carbon atoms or optionally substituted benzyl and R is alkyl with 1 to 6 carbon atoms or optionally substituted phenyl or benzyl and R is alkyl with 1 to 6 carbon atoms, alkenyl with 2 to 6 carbon atoms, alkynyl with 3 to 6 carbon atoms, where the triple bond is not present on the carbon atom attached to the sulfur atom, arylalkyl, alkanoyl with 1 to 6 carbon atoms, arylalkanoyl, aryloxyalkanoyl or arylcarbonyl, each of the R groups being optionally substituted.

European Patent Application Publication No. 44,170 discloses carbapenem derivatives of the formula <°>ni jn , SXN N .3CO ₂ H where R is a hydrogen atom or an organic group bonded via a carbon atom to the carbapenem ring, n is 0 or 1, X is a saturated or unsaturated hydrocarbon radical optionally substituted by a bromine or chlorine atom and R is alkyl of 1 to 6 carbon atoms, alkenyl of 2 to 6 carbon atoms, arylalkyl of 1 to 10 carbon atoms or aryl, any of the R ⁴ groups being optionally substituted. However, the application does not claim any compounds wherein the tetrazole ring is bonded to X via a quaternized nitrogen atom, i.e. a positively charged nitrogen atom not attached to a hydrogen atom.

European Patent Application Publication No. 38,869 mentioned above discloses the synthesis of carbapenem derivatives via intermediates of the general formula wherein R ⁶ R' and R are as defined above, and is a readily removable carboxyl protecting group. Compounds of the formula are also disclosed as intermediates

R ⁶

X

2*

CO ₂ R ^z where X is described as a leaving group.

At the Gordon Research Conference on Medicinal Chemistry held in New London, New Hampshire, August 2-6, 1982, materials were distributed that listed a number of carbapenem antibiotics. Among the compounds listed on page 9 of the materials is a carbapenem with the formula

*®-ch ₃ which differs from the compounds of the present invention in that the quaternized heteroaromatic _t ring in substituent 2 is bonded directly to the sulfur atom instead of to the carbon atom of the alkylene group.

European Published Patent Application No. 50 334 claims carbapenem derivatives of the general formula

NR ¹

II - , SAC-NFVR ²

COOH

7 where R and R are independently selected from the group consisting of hydrogen, alkyl, alkenyl, aryl and arylalkyl, A is a direct single bond connecting sulfur and carbon atoms or A is a cyclic or acyclic linking group selected from the group consisting of substituents 1-2 including alkyl, cycloalkyl, aryl, heteroaryl, or heteroalkyl, R and R which define the carbamimidoyl functional group are independently selected from the group consisting of hydrogen, alkyl and aryl, in addition this carbamimidoyl is characterized by cyclic structures obtained by connecting two nitrogen atoms through substituents and by attachment to the linking group A, further carbamimidinium residues obtained by quaternization of one of the nitrogen atoms of carbamimidoyl are mentioned. On page 12 of this application, possible substituents in position 2 are groups of formula ©

NR ¹ '',

Ί i where r! are hydrogen atoms, substituted and unsubstituted: alkyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, aryl, arylalkyl, heterocyclyl or heterocyclylalkyl and two nitrogen atoms participate in cyclic structures, which are shown in dashed lines

There are no specific claims for any cyclized carbamimidoyl groups containing a quaternized nitrogen atom, but page 22 lists a cyclized carbamimidoyl group of the formula

Based on the above definitions of the R ¹ substituent, we do not believe that European Patent Application No. 50,334 generally claims any of these compounds. Since the wording in said application is so broad with respect to cyclic structures, we incorporate this reference in the present application.

While, as mentioned above, the prior art describes carbapenem derivatives with a substituent at position 2 of the general formula

-S - A - Het where A is an alkylene group and Het represents a heterocyclic or heteroaromatic group there are no claims in which carbapenems are mentioned where Het is a residue of the general formula

where R ⁵ is an optionally substituted aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, heterocyclyl or heterocyclylaliphatic residue and represents a quaternized nitrogen-containing aromatic heterocycle bonded to an alkylene carbon atom through a ring carbon atom. As mentioned above, a carbapenem containing a group of the formula ©,--CH as a substituent in the 2-position

-S-CH,CH,N --— CH, .ch ₃ has been reported, as well as a carbapenem with a quaternized heteroaromatic ring bonded directly to a sulfur substituent.

Despite the huge number of carbapenem derivatives described in the literature, there is a constant need for new carbapenems, as known derivatives can be improved in terms of their spectrum of action, degree of activity, stability and/or toxic side effects.

The present invention relates to a new series of carbapenem derivatives characterized in position 2 by a substituent of the general formula — S-A

Nn ⁵ where A is a straight or branched alkylene with 1 to 6 carbon atoms,

R is an optionally substituted aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, heterocyclyl or heterocyclylaliphatic radical and the radical represents a quaternized nitrogen containing aromatic heterocycle attached to the alkylene group A through a carbon atom in the ring.

The subject of the present invention is a process for the production of carbapenem derivatives of the general formula I

where

Q

R is a hydrogen atom and

R ³ is selected from the group consisting of hydrogen, substituted or unsubstituted: alkyl, alkenyl and alkynyl with up to 10 carbon atoms, cycloalkyl and cycloalkylalkyl with 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moiety, phenyl, arylalkyl, arylalkenyl and arylalkynyl, where the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, where heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical with 5 or 6 members in individual rings and heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic radical with 5 or 6 members in individual rings and where the heteroatom or heteroatoms in the aforementioned moieties are selected from the group consisting of 1 to 4 atoms oxygen, nitrogen or sulfur and the alkyl moieties attached to said heterocyclic moieties have 1 to 6 carbon atoms, where the substituent or substituents of the above-mentioned residues are independently selected from the group comprising alkyl with 1 to 6 carbon atoms, optionally substituted by an amino group, a halogen atom, a hydroxyl or carboxyl group, a halogen atom

-OR ³

O

Iř 3 4 -OCNR R

4 -NR R

NR

3' 4

-so ₂ nr ^j r* * 3 4

-NHCNR R

About 4 -R CNR

-OP(O)(OR ³ )(OR ^{4 * * * *} ) =0

M 3 -OCR

-SR

O

-SR ⁹

-CN

-N

-axis ₃ years

About «

-OS· >1 o

,9

3«

-NR S-R

O

-NR C-NR to ³

4

-NR CO ₂ R

4 where in the above-mentioned substituents the substituents R and R are independently selected from the group consisting of hydrogen, alkyl, alkenyl and alkynyl with 1 to 10 carbon atoms, cycloalkyl, cycloalkylalkyl, and alkylcycloalkyl with 3 to 6 atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moieties, phenyl, arylalkyl, arylalkenyl and arylalkynyl, where the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms and heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, where heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical with 5 or 6 members in individual rings and heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic radical with 5 or 6 members in individual rings and where the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, nitrogen .or sulfur and alkyl moieties attached to heterocyclic moieties have 1 to 6 carbon atoms or R ⁹ and R ⁴ taken together with the nitrogen atom to which at least one is attached may form a five-membered or six-membered nitrogen-containing heterocyclic ring, R 3 18 has the meaning given for R except for the hydrogen atom or R and R together form an alkylidene of 2 to 10 carbon atoms, or an alkylidene of 2 to 10 carbon atoms substituted with hydroxyl,

R ⁵ is substituted or unsubstituted: alkyl, alkenyl and alkynyl with 1 to 10 carbon atoms, cycloalkyl and cycloalkylalkyl with 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moieties, phenyl, arylalkyl, arylalkenyl and arylakynyl, where the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, where heteroaryl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic radical with 5 or 6 members in the individual rings and where the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moieties attached to the above-mentioned heterocyclic moieties have 1 to 6 carbon atoms, where the above-mentioned R radicals are optionally substituted with 1 to 3 substituents independently selected from the group consisting of alkyl with 1 to 6 carbon atoms, optionally substituted with an amino group, a fluorine, chlorine, carboxyl, hydroxyl or carbamoyl atom, a fluorine, chlorine or bromine atom

-OCO ₂ R ³

-OCOR ³

-oconr ³ r ⁴ o

H ₉ -OS-R »1

O

4 -NR R

4

R ^J C0NR 3 4

-NR CO ₂ R

31| q -NR ^J SR ^y

II

O

-SR

About 0

-V- R ⁹

-SO ₃ ^r3

-co ₂ r ³

4

-CONR R

-CN phenyl optionally substituted with 1 to 3 fluorine, chlorine, bromine, alkyl with 1 to 6 carbon atoms, OR -HR^R^, -SO^R^, -CO^R^ or -CONR^R^, where R^ and and R^ in the R^ substituents have the meaning given above, or R° may be attached at another position of the ring so as to form a fused heterocyclic or heteroaromatic ring, which may further contain other heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur, is a hydrogen atom, substituted and unsubstituted alkyl, alkenyl and alkynyl with 1 to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl with 3 to 6 carbon atoms in the cycloalkyl ring with 1 to 6 carbon atoms in the alkyl moieties, spirocycloalkyl with 3 to 6 carbon atoms, phenyl, arylalkyl, arylalkenyl and arylalkynyl, where the aryl moiety is phenyl and the aliphatic part has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, where heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical with 5 or 6 members in individual rings and heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic radical with 5 or 6 members in individual rings and where the heteroatom or heteroatoms in the above-mentioned heterocyclic parts are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms, and the alkyl parts attached to these heterocyclic parts have 1 to 6 carbon atoms, where the substituent or substituents of the above-mentioned radicals are selected from the group consisting of amino, mono-, di-trialkylamino, hydroxyl, alkoxy, mercapto, alkylthio, phenylthio, sulfamoyl, amidino, guanidino, nitro, chloro, bromo, fluorine, cyano and carboxyl and where the alkyl moieties in the above substituents have 1 to 6 carbon atoms,

A is a straight or branched alkylene having 1 to 6 carbon atoms,

R is a hydrogen atom, an anionic charge or a common readily cleavable carboxyl protecting group selected from the group comprising benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloroethyl, trialkylsilyl, where the alkyls have 1 to 4 carbon atoms, phenacyl, ρ-methoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridiylmethyl, alkyl with 1 to 6 carbon atoms, pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, methoxymethyl and preferably p-nitrobenzyl, wherein if R is a hydrogen atom or a protecting group, an ion is also present and the group represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocyclic radical containing at least one nitrogen atom in the ring and a total of 1 to 4 nitrogen, oxygen or sulfur atoms and attached to A via a carbon atom in the ring and having the nitrogen atom in the ring which is quaternized by the group R^ and their pharmaceutically acceptable salts, characterized in that the intermediate of general formula II is reacted

where r1, r8, ^r15 _and have the meaning given above, '

R is a common readily cleavable carboxyl protecting group selected from the group consisting of benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloroethyl, trialkylsilyl, where the alkyls have 1 to 4 carbon atoms, phenacyl, p-methoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl, alkyl with 1 to 6 carbon atoms, pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, methoxymethyl and preferably p-nitrobenzyl and _t represents a mono-, bi- or polycyclic aromatic heterocyclic radical containing a quaternizable nitrogen atom in the ring, said ring being attached to A via a carbon atom in the ring, in an inert organic solvent with an alkylating agent of the general formula

5 where R is as defined above and X is a common leaving group such that the R group quaternizes the nitrogen atom in the ring of the substituent in the intermediate of general formula II to form a compound of general formula I

S-A z- ©

COOR ² '"15

N-r5 , have the meaning given above and optionally the anion X is replaced by another anion and optionally the carboxyl protecting group R is 2' removed to form an unprotected compound of general formula I or pharmaceutically acceptable salts thereof.

Compounds of formula X are effective antibacterial agents or intermediates useful in the preparation of such agents.

The present invention also relates to a process for the preparation of the novel carbapenem derivatives described above and pharmaceutical compositions containing biologically active carbapenem derivatives in combination with pharmaceutically acceptable carriers or diluents.

The novel compounds of general formula I above contain a carbapenem nucleus

and may be referred to as derivatives of l-carba-2-penem-3-carboxylic acid. Alternatively, the compounds may be considered to have the basic formula

and referred to as 7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylic acid derivatives. While the present invention encompasses compounds in which the relative stereochemistry of the protons at the 5 and 6 positions is cis as well as trans, preferred compounds have the 5R,6S (trans) stereochemistry of thienamycin.

Compounds of formula I may be unsubstituted in the 6-position or substituted with the groups 8-1 as set out above for other carbapenem derivatives. More specifically, R may be hydrogen and R may be hydrogen or a substituent other than hydrogen, as set out for example in European Patent Application Publication No. 38,869 (see definition of Rg).

1

Alternatively, R and R together may form an alkylidene of 2 to 10 carbon atoms or an alkylidene of 2 to 10 carbon atoms substituted, for example, by hydroxyl.

Compounds of formula I may also be unsubstituted in position 1 (R1 = h) or substituted with the substituents listed above for other carbapenem derivatives.

More specifically, R may be a hydrogen atom or a substituent other than a hydrogen atom as disclosed, for example, in European Patent Application Publication No. 54,917 (as defined in R or R ) or in U.S. Patent No. 4,350,631.

Preferred substituents of R^ other than hydrogen are alkyl of 1 to 6 carbon atoms, preferably methyl, phenyl and phenylalkyl of 1 to 6 carbon atoms in the alkyl. Substituents other than hydrogen may be in either the alpha or beta configuration and it is understood that the present invention includes individual alpha and beta isomers as well as mixtures thereof. The most preferred 1-substituted compounds are those having the beta configuration, especially those having beta-methyl.

15

For definitions for R , R , and R expressions

a) aliphatic alkyl, alkenyl and alkynyl may have a straight or branched chain with 1 to 10 carbon atoms, preferably 1 to 6 carbon atoms, most preferably 1 to 4 carbon atoms, if <

247195 20 these are part of another substituent, for example cycloalkylalkyl or heteroarylalkyl or arylalkenyl, then alkyl, alkenyl and alkynyl preferably contain 1 to 6, preferably 1 to 4, carbon atoms;

b) heteroaryl includes mono-, bi- and polycyclic aromatic heterocyclic groups with 1 to 4 oxygen, nitrogen or sulfur atoms, preferably five- or six-membered heterocyclic rings, such as thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, fetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyrilazinyl, pyrrolyl, pyrazolyl, etc.;

c) heterocyclyl includes mono-, bi- and polycyclic saturated or unsaturated non-aromatic heterocyclic groups containing 1 to 4 oxygen, nitrogen or sulfur atoms, preferably five- or six-membered heterocyclic rings, such as morpholinyl, piperazinyl, piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrvolinyl, pyrrolidinyl and the like;

d) the term halogen includes chlorine, bromine, fluorine and iodine atoms and preferably chlorine, bromine or fluorine.

The term readily removable carboxyl protecting group means a known ester group which is used to protect the carboxyl group during the chemical reaction steps described below and which can optionally be removed by methods which do not result in any destruction of the remaining parts of the molecule, for example by chemical or enzymatic hydrolysis, reaction with chemical reducing agents under mild conditions, irradiation with ultraviolet radiation or catalytic hydrogenation.

Examples of such ester protecting groups are benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloroethyl, trialkylsilyl, where the alkyls have 1 to 4 carbon atoms, phenacyl, p-methoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl and alkyl with 1 to 6 carbon atoms, such as methyl, ethyl or tert-butyl.

These protecting groups include those that are hydrolyzed under physiological conditions, such as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethyl.

A particularly preferred carboxyl protecting group is p-nitrobenzyl, which is readily removed by catalytic hydrogenolysis.

The pharmaceutically acceptable salts mentioned above include non-toxic salts with acids, for example salts with mineral acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid and the like, and salts with organic acids such as maleic acid, acetic acid, citric acid, succinic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, ascorbic acid, lactic acid, gluconic acid and malonic acid.

Compounds of general formula X in the form of salts with acids can be written as

R = H or a protecting group where x" is the anion of the acid. The anion x may be chosen to provide a pharmaceutically acceptable salt for therapeutic applications, but in the case of intermediates of compounds of formula I the anion X may also be a toxic anion.

In this case, the ion can then be removed or substituted with a pharmaceutically acceptable anion to form an active end product for therapeutic use. If acidic or basic groups are present in the R or R groups or in the residue

The present invention also includes suitable basic or acidic salts of these functional groups, for example, acid addition salts in the case of a basic group and metal salts (e.g., sodium, potassium, calcium, and aluminum), ammonium salts, and salts with non-toxic amines (e.g., trialkylamines, procaine, dibenzylamine, 1-ephenamine, N-benzyl-beta-phenylethylamine, N,N'-dibenzylethylenediamine, etc.) in the case of an acidic group.

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

Preferred compounds of the present invention are compounds of formula I, wherein R is hydrogen and R^ is hydrogen, CH^Cl^ ^- ,

CH.

'CH-,

CH. OH

CH.

OH or CH ₃ CHZ of this subgroup, the preferred compounds are those where R is

OH the most preferred compounds are those having the absolute configuration 5R, 6S, 8R.

Other preferred compounds of the present invention are compounds of general formula I, wherein R and R together form an alkylidene radical of the formula "C=

CH.'

The alkylene radical (i.e., substituent A”) in a compound of formula I may be straight-chain or branched and may contain from 1 to 6 carbon atoms. Preferred compounds are those wherein A is “( ^CH 2^n” ^ř where n is 1 or 2, and a particularly preferred embodiment includes those compounds,

The alkylene portion of molecule A is attached via a ring carbon atom to an N-substituted quaternized aromatic heterocycle of the general formula wherein the r5 substituent is preferably optionally substituted alkyl of 1 to 6 carbon atoms, alkenyl of 2 to 10 carbon atoms, alkynyl of 2 to 10 carbon atoms, cycloalkyl of 3 to 6 carbon atoms, cycloalkylalkyl of 6 carbon atoms in the cycloalkyl and 1 to 6 carbon atoms in the alkyl moiety, phenyl, phenylalkyl of 1 to 6 carbon atoms in the alkyl moiety, phenylalkenyl of 2 to 6 carbon atoms in the alkenyl moiety, phenylalkynyl of 2 to 6 carbon atoms in the alkynyl moiety, heteroaryl, heteroarylalkyl, where the alkyl moiety has 1 to 6 carbon atoms, heterocyclyl or heterocyclylalkyl, where the alkyl moiety has 1 to 6 carbon atoms carbon.

Heteroaryl (or the heteroaryl part of a heteroarylalkyl) as an R substituent can be a mono-, bi-, or polycyclic, aromatic heterocyclic group containing 1 to 4 oxygen, nitrogen, or sulfur atoms, preferably a five- or six-membered heterocyclic ring such as thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, tetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, and pyrazolyl.

C

Heterocyclyl (or the heterocyclic portion of heteroalkylalkyl) as an R substituent can be a mono-, bi- or polycyclic saturated or unsaturated non-aromatic heterocyclic group containing 1 to 4 oxygen, nitrogen, or sulfur atoms, preferably five- or six-membered heterocyclic rings, such as morpholinyl, piperazinyl, piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrrolinyl, and pyrrolidinyl.

The substituent R ³ may be optionally substituted with 1 to 3 substituents selected independently from the group consisting of

a) alkyl with 1 to 6 carbon atoms, optionally substituted with 1 to 3 substituents from the group consisting of amino, fluorine, chlorine, carboxyl, hydroxyl or carbamoyl,

b) an atom of fluorine, chlorine or bromine,

o) -QR ³ ,

d) -OCO ₂ R ³ ,

e) -OCOR ³ ,

f) -OCONR ³ R ⁴ , li q

g) -OS-R, á

h) -oxo,

4

i) -NR R ,

j) r ³ conr ⁴ -,

k) -nr ³ co ₂ r ⁴ ,

l) -NR ³ CONR ³ R ⁴ ,

Oh my god

m) -NR S-R , li

O

n) -SR ³ ,

o) -SOR ⁹ ,

About ¹¹ 9

p) -S-R ,

q) -SO ₃ R ³ ,

r) -CO ₂ R ³ ,

s) -CONR ³ R ⁴ ,

t) -CN or

u) phenyl, optionally substituted with 1 to 3 substituents independently selected from the group consisting of fluorine, chlorine, bromine, alkyl with 1 to 6 carbon atoms, -OR , 3 4 3 3 3 4

-NR R , -SO^R , -CC^R or -CONR R , where with respect to the above-mentioned substituents R 5 the groups R ^ and R ^ are independently selected from the group comprising a hydrogen atom, alkyl, alkenyl, and alkynyl with 1 to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl with 3 to 6 carbon atoms in the cycloalkyl part and 1 to 6 carbon atoms in the alkyl part of the molecule, phenyl, arylalkyl, arylalkenyl and arylalkynyl, where the aryl part is phenyl and the aliphatic part has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, hetero cyclyl and heterocyclylalkyl, where the heteroaryl and heterocyclyl group or part of the group has the meaning defined above for R ° and the alkyl parts attached to these hetero3 4 cyclic parts have 1 to 6 carbon atoms, or R and R together with the nitrogen atom to which at least one is attached, may form a five-membered or six-membered nitrogen containing heterocyclic (as defined above for R ) ring and R is as defined above for except that it is not a hydrogen atom. The most preferred substituent '

R is alkyl of 1 to 6 carbon atoms, especially methyl.

Additionally, the R substituent together with another atom in the ring portion of the molecule may form a fused heterocyclic or heteroaromatic ring, wherein this ring may contain an additional preferably 1 or 2 heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur.

For example

Group

or preferably represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocycle containing at least one ring nitrogen atom and 0 to 5 other heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur, said heterocyclic ring being attached to A through a ring carbon atom and having the ring nitrogen atom quaternized by the group R.

The heteroaromatic —ί- N— ring may be optionally substituted on available carbon atoms in the ring with preferably 1 to 5 substituents, most preferably 1 to 3 substituents independently selected from the group consisting of alkyl with 1 to 4 carbon atoms, alkyl with 1 to 4 carbon atoms substituted with preferably 1 to 3 hydroxy groups, amino groups, alkylamino groups with 1 to 4 carbon atoms, dialkylamino groups with 1 to 4 carbon atoms in each of the alkyls, alkoxy groups with 1 to 4 carbon atoms, carboxy group, halogen atoms (meaning chlorine, bromine, fluorine or iodine atom, preferably chlorine, bromine or fluorine) or sulfo groups, cycloalkyl with 3 to 6 carbon atoms, cycloalkylalkyl with 3 to 6 carbon atoms in the cycloalkyl part and 1 to 4 carbon atoms in the alkyl part, optionally substituted with 1 to 3 substituents mentioned above in connection with alkyl with 1 to 4 carbon atoms, alkoxy with 1 to 4 carbon atoms, alkylthio group with 1 to 4 carbon atoms, aminodialkylamino group with 1 to 4 carbon atoms, halogen atom, alkanoylamino group with 1 to 4 carbon atoms, alkanoyloxy group with 1 to 4 carbon atoms, carboxyl, sulfo group, -C-O-alkyl group with 1 to 4 carbon atoms in alkyl, hydroxy group, amidino group, guanidino group, phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group consisting of amino group, halogen atom, hydroxyl, trifluoromethyl, alkyl with 1 to 4 carbon atoms, alkoxy with 1 to 4 carbon atoms, alkylamino group with 1 to 4 carbon atoms, dialkylamino group with 1 to 4 carbon atoms in each of alkyls, carboxy groups and sulfo groups; phenylalkyl of 1 to 4 carbon atoms, wherein the phenyl portion may be optionally substituted with 1 to 3 substituents listed above in connection with phenyl and the alkyl portion may be optionally substituted with 1 to 3 substituents listed above in connection with alkyl of 1 to 4 carbon atoms and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl portion attached to the heteroarylalkyl has 1 to 6 carbon atoms, wherein these heteroaryl and heteroarylalkyl groups are optionally substituted in the heterocyclic ring with 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, alkylamino of 1 to 4 carbon atoms, dialkylamino of 1 to 4 carbon atoms carbon in each of the alkyl, carboxy and sulfo groups and the alkyl portion may be substituted with 1 to 3 substituents selected from the group consisting of hydroxyl, amino, alkylamino with 1 to 4 carbon atoms, dialkylamino with 1 to 4 carbon atoms in each of the alkyls, alkoxy with 1 to 4 carbon atoms, carboxyl, halogen and sulfo.

Additionally, the nitrogen atoms present in the ring (except for the quaternized nitrogen atom) may be substituted with 1 to 3 substituents independently selected from the group consisting of alkyl with 1 to 4 carbon atoms, alkyl with 1 to 4 carbon atoms preferably substituted with 1 to 3 hydroxy groups, amino groups, alkylamino groups with 1 to 4 carbon atoms, dialkylamino groups with 1 to 4 carbon atoms in each of the alkyls, alkoxyl groups with 1 to 4 carbon atoms, carboxy groups, halogen atoms or sulfo groups, cycloalkyl with 3 to 6 carbon atoms, cycloalkylalkyl with 3 to 6 carbon atoms in the cycloalkyl part and 1 to 4 carbon atoms in the alkyl part, optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms, phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group consisting of from the group consisting of amino, halogen, hydroxyl, trifluoromethyl, alkyl with 1 to 4 carbon atoms, alkoxy with 1 to 4 carbon atoms, alkylamino with 1 to 4 carbon atoms, dialkylamino with 1 to 4 carbon atoms, carboxy and sulfo, phenylalkyl with 1 to 4 carbon atoms in the alkyl portion, wherein the phenyl portion may be optionally substituted with 1 to 3 substituents listed above in connection with phenyl and the alkyl portion may be optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl portion associated with the heteroarylalkyl has 1 to 6 carbon atoms, wherein the heteroaryl and heteroarylalkyl groups are optionally substituted in the heterocyclic ring 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluoromethyl, alkyl with 1 to 4 carbon atoms, alkoxy with 1 to 4 carbon atoms, alkylamino with 1 to 4 carbon atoms, dialkylamino with 1 to 4 carbon atoms, carboxyl and sulfo and the alkyl part is substituted with 1 to 3 substituents selected from the group consisting of hydroxyl, amino, alkylamino with 1 to 4 carbon atoms, dialkylamino with 1 to 4 carbon atoms in each of the alkyls, alkoxy with 1 to 4 carbon atoms, carboxyl, halogen and sulfo.

is alkyl with 1 to 6 preferably compounds, -CH ₂ " and where

The most preferred substituent on the carbon atom and the nitrogen atom in the ring is carbon atoms, especially methyl.

The above-described preferred embodiment of the present invention includes those wherein A is -(CH 2 )-, wherein n is 1 or 2, most preferably wherein A is

8

a) R and R together form ch;

or

1

b) R is a hydrogen atom and R is a hydrogen atom, CH ₂ CH ₂ -,

CHCHCH _O OH I

Cor

OH

I ch ₃ ch-.

_CHO

CH _O ,1 ·

Particularly preferred are compounds where R1 is hydrogen and ^R2 is

OH

I ch ₃ -chespecially compounds with absolute configuration 5R, 6S, 8R.

In a preferred embodiment, the group represents an aromatic five- or six-membered nitrogen-containing heterocyclic ring containing 0 to 3 further heteroatoms selected from the group consisting of an oxygen, sulfur or nitrogen atom. This aromatic heterocycle may, where possible, be connected to another ring, which may be an unsaturated or saturated carbocyclic ring, preferably a carbocyclic ring with 4 to 7 carbon atoms, an aromatic carbocyclic ring preferably phenyl, a 4 to 7 membered heterocyclic ring (saturated or unsaturated) containing 1 to 3 heteroatoms selected from the group consisting of an oxygen, sulfur, nitrogen atom or a group NrH, where the hydrogen atom is an alkyl with 1 to carbon atoms, optionally substituted with 1 to 2 substituents independently selected from 3 3 4 3 from the group consisting of -OR , -NR R , -CO _? R , oxo group, phenyl, a fluorine, chlorine, bromine atom,

3 4 group -SO ₃ R and -CONR R or phenyl, optionally substituted with 1 to 3 substituents independently

3 4 on themselves selected from the group consisting of alkyl with 1 to 6 carbon atoms, OR , -NR R , a fluorine, chlorine, bromine atom, -SO,R^, -CO-R^ and -CONR^R ⁴ , where R ³ and R ⁴ in these R ⁴⁴ substituents have the meaning defined in connection with the substituent R or a five- or six-membered heterocyclic ring containing 1 to 3 heteroatoms selected from the group consisting of an oxygen, sulfur, nitrogen or NR'*' ⁴ , where R ¹⁴ has the meaning given above.

The five- or six-membered aromatic quaternized ring or, where appropriate, the carbocyclic, heterocyclic or heteroaromatic ring attached thereto, or both of these rings may be optionally substituted on available atoms in the ring, preferably up to a total of five substituents on the overall ring system, the substituents being as set out above in connection with the group

In the preferred embodiment described _{above, preferred compounds are those where A is -CH2} ⁾ where n is 1 or 2, most preferably those where A is -CH2, and where

8

a) R and R together form a group

CHI

1

b) R is a hydrogen atom and R is a hydrogen atom, CHjC^-,

CH, ,CH-,

CH, OH :c _? H

CH,

CH,

1

Particularly preferred are compounds where R is a hydrogen atom and R is, in particular, those compounds which have the absolute configuration 5R, 6S, 8R.

Another preferred embodiment of the present invention includes compounds of general formula I, wherein

NR ⁶ represents a residue selected from the group consisting of

a) the rest of the formula

^R5

R ⁷ _R 10 where R ⁶ , R ⁷ and R ¹⁸ are independently selected from the group comprising a hydrogen atom, an alkyl with 1 to 4 carbon atoms, optionally substituted with 1 to 3 hydroxy groups, alkylamino groups with 1 to 4 carbon atoms, dialkylamino groups with 1 to 4 carbon atoms in each of the alkyls, alkoxyls with 1 to 4 carbon atoms, amino groups, sulfo groups, carboxy groups or halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), cycloalkyl with 3 to 6 carbon atoms, alkoxyl with 1 to 4 carbon atoms, alkylthio group with 1 to 4 carbon atoms, amino group, alkylamino group with 1 to 4 carbon atoms, dialkylamino group with 1 to 4 carbon atoms in each of the alkyls, halogen atom, (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), alkanoylamino group with 1 to 4 carbon atoms, alkanoyloxy group with 1 to 4 carbon atoms, carboxy group, -C-O alkyl with 1 to 4 carbon atoms, hydroxyl,

O amidino, guanidino, phenyl, phenyl substituted with one, two or three amino groups, halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyls, trifluoromethyls, alkyls with 1 to 4 carbon atoms or alkoxys with 1 to 4 carbon atoms, phenylalkyls with 1 to 4 carbon atoms in the alkyl moiety, where the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in connection with phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms and heteroaryl and heteroarylalkyl, where the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms, the alkyl moiety attached to this heteroalkyl moiety has 1 to 6 carbon atoms or where two of the substituents R®, R ⁷ or R ³⁸ together may be joined to form a saturated fused carbocyclic ring, a fused aromatic carbocyclic ring, a fused non-aromatic, heterocyclic ring or a fused heteroaromatic ring, said fused ring being optionally substituted with 1 or 2 substituents as defined above for R ⁸ , R ⁷ and R ³⁸ ,

b) formula residues

^R5

R ⁵ '“her or

N

R ⁵ íL optionally substituted on a carbon atom by one to three substituents independently selected from the group comprising alkyl with 1 to 4 carbon atoms, alkyl with 1 to 4 carbon atoms, preferably substituted with 1 to 3 hydroxy groups, alkylamino groups with 1 to 4 carbon atoms, sulfo groups, dialkylamino groups with 1 to 4 carbon atoms in each of the alkyls, alkoxy groups with 1 to 4 carbon atoms, amino groups, carboxy groups or halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), cycloalkyl with 3 to 6 carbon atoms, alkoxyl with 1 to 4 carbon atoms, alkylthio group with 1 to 4 carbon atoms, amino group, alkylamino group with 1 to 4 carbon atoms, dialkylamino group with 1 to 4 carbon atoms in each of the alkyls, halogen atom (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), alkanoylamino group with 1 to 4 carbon atoms, alkanoyloxy group with 1 to 4 carbon atoms, carboxyl, -CO-alkyl group with 1 to 4 carbon atoms in the alkyl, hydroxyl, aminidino group, guanidino group, phenyl, phenyl substituted with one, two or three amino groups, halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyls, trifluoromethyls, alkyls with 1 to 4 carbon atoms or alkoxys with 1 to 4 carbon atoms, phenylalkyl with 1 to 4 carbon atoms, where the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in connection with phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 up to 4 carbon atoms and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen, or sulfur atoms and the alkyl moiety linked to the heteroarylalkyl moiety has 1 to 6 carbon atoms or is optionally substituted to form a fused carbocyclic, heterocyclic, or heteroaromatic ring, optionally substituted with 1 or 2 substituents as defined above?

c) leftovers

^R5

^R5

^R5

R ⁵ i\r tm

R ⁵ .

or

R5

optionally substituted on a carbon atom by one to two substituents independently selected from the group comprising alkyl with 1 to 4 carbon atoms, alkyl with 1 to 4 carbon atoms, preferably substituted with 1 to 3 hydroxy groups, alkylamino groups with 1 to 4 carbon atoms, sulfo groups, dialkylamino groups with 1 to 4 carbon atoms in each of the alkyls, alkoxy groups with 1 to 4 carbon atoms, amino groups, carboxy groups or halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine fluorine or bromine), cycloalkyl with 3 to 6 carbon atoms, alkoxyl with 1 to 4 carbon atoms, alkylthio group with 1 to 4 carbon atoms, amino group, alkylamino group with 1 to 4 carbon atoms, dialkylamino group with 1 to 4 carbon atoms in each of the alkyls, halogen atom (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), alkanoylamino group with 1 to 4 carbon atoms, alkanoyloxy group with 1 to 4 carbon atoms, carboxyl, -C-O-alkyl group with 1 to 4 carbon atoms in the alkyl, hydroxy1, amidino group, guanidino group,

O phenyl, phenyl substituted with one, two or three amino groups, halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyls, trifluoromethyls, alkyls with 1 to 4 carbon atoms or alkoxys with 1 to 4 carbon atoms, phenylalkyl with 1 to 4 carbon atoms, where the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in connection with phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms, and heteroaryl or heteroarylalkyl, where the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety linked to the heteroarylalkyl moiety has 1 to 6 carbon atoms or is optionally substituted to form a fused carbocyclic, heterocyclic or a heteroaromatic ring, optionally substituted with 1 or 2 substituents as defined above;

d) residues of formulas >IM

JM,5 n

rV

or

optionally substituted on a carbon atom by a substituent independently selected from the group consisting of alkyl with 1 to 4 carbon atoms, alkyl with 1 to 4 carbon atoms, preferably substituted with 1 to 3 hydroxy groups, alkylamino groups with 1 to 4 carbon atoms, sulfo groups, dialkylamino groups with 1 to 4 carbon atoms in each of the alkyls, alkoxy groups with 1 to 4 carbon atoms, amino groups, carboxy groups or halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), cycloalkyl with 3 to 6 carbon atoms, alkoxyl with 1 to 4 carbon atoms, alkylthio group with 1 to 4 carbon atoms, amino group, alkylamino group with 1 to 4 carbon atoms, dialkylamino group with 1 to 4 carbon atoms in each of the alkyls, halogen atom (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), alkanoylamino group with 1 to 4 carbon atoms, alkanoyloxy group with 1 to 4 carbon atoms, carboxyl, -g-O-alkyl group with 1 to carbon atoms in the alkyl, hydroxyl, amidino group, guanidino group, phenyl, phenyl substituted with one, two or three amino groups, halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyls, trifluoromethyls, alkyls with 1 to 4 carbon atoms or alkoxys with 1 to 4 carbon atoms, phenylalkyl with 1 to 4 carbon atoms, where the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in connection with phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms, and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety attached to the heteroarylalkyl moiety has 1 to 6 carbon atoms,

e) formula residues

where X is oxygen, sulfur or NR, where R is alkyl with 1 to 4 carbon atoms, alkyl with 1 to 4 carbon atoms substituted with 1 to 3 hydroxy groups, amino groups, alkylamino groups with 1 to 4 carbon atoms, dialkylamino groups with 1 to 4 carbon atoms in each of the alkyls, alkoxy groups with 1 to 4 carbon atoms, carboxy groups, halogen atoms or sulfo groups, cycloalkyl with 3 to 6 carbon atoms, cycloalkylalkyl with 3 to 6 carbon atoms in the cycloalkyl part and 1 to 4 carbon atoms in the alkyl part, optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms, phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group consisting of amino group, halogen atom, hydroxyl, trifluoromethyl, alkyl with 1 to 4 atoms carbon, alkoxy with 1 to 4 carbon atoms, alkylamino with 1 to 4 carbon atoms, dialkylamino with 1 to 4 carbon atoms in each of the alkyls, carboxyl and sulfo, phenylalkyl with 1 to 4 carbon atoms in the alkyl portion, where phenyl may be optionally substituted with 1 to 3 substituents listed above in connection with phenyl and the alkyl portion may be optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms and heteroaryl and heteroarylalkyl, where the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl portion attached to the heteroarylalkyl has 1 to 6 carbon atoms, wherein the heteroaryl and heteroarylalkyl groups are optionally substituted in the heterocyclic ring with 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluoromethyl, alkyl with 1 to 4 carbon atoms, alkoxy with 1 to 4 carbon atoms, alkylamino with 1 to 4 carbon atoms, dialkylamino with 1 to 4 carbon atoms in each of the alkyls, carboxy and sulfo and in the alkyl part 1 to 3 substituents selected from the group consisting of hydroxyl, amino, alkylamino. a dialkylamino group with 1 to 4 carbon atoms in each of the alkyls, an alkoxy group with 1 to 4 carbon atoms, a carboxyl, a halogen atom and a sulfo group, said heteroaromatic residue is optionally substituted on a carbon atom by one or more substituents independently selected from the group comprising alkyl with 1 to 4 carbon atoms, alkyl with 1 to 4 carbon atoms preferably substituted with 1 to 3 hydroxy groups, amino groups, alkylamino groups with 1 to 4 carbon atoms, dialkylamino groups with 1 to 4 carbon atoms in each of the alkyls, alkoxy groups with 1 to 4 carbon atoms, sulfo groups, carboxy groups or halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), cycloalkyl with 3 to 6 carbon atoms, alkoxy with 1 to 4 carbon atoms, alkylthio group with 1 to 4 carbon atoms, amino group, alkylamino group with 1 to 4 carbon atoms, dialkylamino group with 1 to 4 carbon atoms, in each of the radicals, a halogen atom (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), alkanoylamino group with 1 to 4 carbon atoms, alkanoyloxy group with 1 to 4 carbon atoms, carboxy group, -C-O-alkyl with 1 to 4 carbon atoms in alkyl, hydroxyl, amidino group, guanidino group, phenyl, phenyl substituted with one, two or three substituents, such as amino group, halogen atom (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyl, trifluoromethyl, alkyl with 1 to 4 carbon atoms or alkoxy with 1 to 4 carbon atoms, phenylalkyl with 1 to 4 carbon atoms, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents as defined above in relation to phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents as defined above in relation to alkyl having 1 to 4 carbon atoms, heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety attached to said heteroarylalkyl moiety has 1 to 6 carbon atoms or is optionally substituted to form an attached carbocyclic, heterocyclic or heteroaromatic ring, optionally substituted with 1 or 2 substituents as defined above;

f) formula residues

R5

\Φ

N —X —(ΓΊΙ© c hk^N-R"

Φ ϋ π—NR ⁵ i|j_ ^Ν ?χ^

X—N—R® or N—NR ⁵ where X is an oxygen, sulfur or NR atom, where R is alkyl with 1 to 4 carbon atoms, alkyl with 1 to 4 carbon atoms substituted with 1 to 3 hydroxy groups, amino groups, alkylamino groups with 1 to 4 carbon atoms, dialkylamino groups with 1 to 4 carbon atoms in each of the alkyls, alkoxy groups with 1 to 4 carbon atoms, carboxy groups, halogen atoms or sulfo groups cycloalkyl with 3 to 6 carbon atoms in the cycloalkyl part and 1 to 4 carbon atoms in the alkyl part, optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms, phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group comprising amino group, halogen atom, hydroxyl, trifluoromethyl, alkyl with 1 to 4 carbon atoms, alkoxy with 1 to 4 carbon atoms, alkylamino with 1 to 4 carbon atoms, dialkylamino with 1 to 4 carbon atoms in each of the alkyls, carboxyl and sulfo, phenylalkyl with 1 to 4 carbon atoms in the alkyl portion, where phenyl may be optionally substituted with 1 to 3 substituents listed above in connection with phenyl and the alkyl portion may be optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms and heteroaryl and heteroarylalkyl, where the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl portion attached to the heteroarylalkyl has 1 to 6 carbon atoms, wherein the heteroaryl and heteroarylalkyl groups are optionally substituted in the heterocyclic ring with 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluoromethyl, alkyl with 1 to 4 carbon atoms, alkylamino group with 1 to 4 carbon atoms, dialkylamino group with 1 to 4 carbon atoms in each of the alkyls, carboxy group and sulfo group and in the alkyl part 1 to 3 substituents selected from the group comprising hydroxyl, amino group, alkylamino group with 1 to 4 carbon atoms, dialkylamino group with 1 to 4 carbon atoms in each of the alkyls, alkoxyl with 1 to 4 carbon atoms, carboxyl, halogen atom and sulfo group, said heteroaromatic residue is optionally substituted on the carbon atom by one or more substituents independently selected from the group comprising alkyl with 1 to 4 carbon atoms, alkyl with 1 to 4 carbon atoms preferably substituted with 1 to 3 hydroxyl groups, amino groups., alkylamino groups with 1 to 4 carbon atoms, dialkylamino groups with 1 to 4 carbon atoms in each of the radicals, alkoxy groups with 1 to 4 carbon atoms, sulfo groups, carboxy groups or halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), cycloalkyl with 3 to 6 carbon atoms, alkoxy with 1 to 4 carbon atoms, alkylthio group with 1 to 4 carbon atoms, amino group, alkylamino group with 1 to 4 carbon atoms, dialkylamino group with 1 to 4 carbon atoms in each of the alkyls, halogen atom (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), alkanoylamino group with 1 to 4 carbon atoms, alkanoyloxy group with 1 to 4 carbon atoms, carboxy group, -CO-alkyl with 1 to 4 carbon atoms in the alkyl, hydroxyl, amidino group, guanidino group, phenyl, phenyl substituted with one, two or three substituents such as amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyl, trifluoromethyl, alkyl with 1 to 4 carbon atoms or alkoxy with 1 to 4 carbon atoms, phenylalkyl, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in connection with phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms, heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety attached to said heteroarylalkyl moiety has 1 to 6 carbon atoms;

g) formula residues

n_n-r II I c ’

Nx^NR ⁵ „ $ _r 5_n-~ N — R

II I

N^N or n-n-R

where R is alkyl with 1 to 4 carbon atoms, alkyl with 1 to 4 carbon atoms substituted with 1 to 3 hydroxy groups, amino groups, alkylamino groups with 1 to 4 carbon atoms, dialkylamino groups with 1 to 4 carbon atoms in each of the carbons, alkoxy groups with 1 to 4 carbon atoms, carboxy groups, halogen atoms or sulfo groups, cycloalkyl with 3 to 6 carbon atoms, cycloalkylalkyl with 3 to 6 carbon atoms in the cycloalkyl part and 1 to 4 carbon atoms in the alkyl part, optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms, phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group consisting of amino group, halogen atom, hydroxyl, trifluoromethyl, alkyl with 1 to 4 carbon atoms, alkoxy with 1 to 4 carbon atoms, alkylamino group with 1 to 4 carbon atoms, dialkylamino group with 1 to 4 carbon atoms in each of the alkyls, carboxy group and sulfo group, phenylalkyl with 1 to 4 carbon atoms in the alkyl part, where the phenyl part may be optionally substituted with 1 to 3 substituents listed above in connection with phenyl and the alkyl part may be optionally substituted with 1 to 3 substituents listed above in connection with alkyl with 1 to 4 carbon atoms, heteroaryl and heteroarylalkyl, where the heteroatom and heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl part connected to the heteroarylalkyl has 1 to 6 carbon atoms, said heteroaryl and heteroarylalkyl groups are optionally substituted in the heterocyclic ring with 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluoromethyl, alkyl with 1 to 4 carbon atoms, alkoxy with 1 to 4 carbon atoms, alkylamino with 1 to 4 carbon atoms, dialkylamino with 1 to 4 carbon atoms in each of the alkyls, carboxy and sulfo and the alkyl part is substituted with 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, alkylamino with 1 to 4 carbon atoms, dialkylamino with 1 to 4 carbon atoms in each of the alkyls, alkoxy with 1 to 4 carbon atoms, carboxyl, halogen and sulfo.

The substituents R and R may also form a fused heterocyclic or heteroaromatic ring.

Of the above-described preferred embodiments of the present invention, those compounds wherein A is ^CH 2*n ^and n are preferred, and those wherein A is Cl^" ^and where

8

a) R and R together form ^H0CH 2 or

1

b) R is a hydrogen atom and R is a hydrogen atom, CH^Cl^-

Cil., 3 ^CH3- OH OH C or _CH3CH- . ^CH r^ ^ch 3 Especially advantageous are compounds. where R ⁸ is a hydrogen atom and is

OH (

CH ₃ -CH, especially compounds of absolute configuration 5R, 6S, 8R.

A particularly preferred embodiment of the present invention includes compounds of formula I, wherein the residue of the formula represents

where r6, ^r7 and are independently selected from the group consisting of hydrogen, alkyl with 1 to 4 carbon atoms, alkoxy with 1 to 4 carbon atoms, carboxyl and carbamoyl and ^R3 has the meaning given above and is preferably alkyl with 1 to 6 carbon atoms, most preferably methyl.

Of the above-described preferred embodiments of the present invention, those compounds are preferred where A is -(CH ₂ ) _n ~, where n is 1 or 2, more preferred where A is -CH ₂ “ and where

a) R and R together form

BOY

C=

CH or

1

b) R° is a hydrogen atom and R is a hydrogen atom, CH^CH^,

CHCH-, ^CH 3 of the CH^CHCHCHCHl

1

Particularly preferred are compounds where R is hydrogen and R is

OH especially those compounds that have the absolute configuration 5R, 6S, 8R.

Another preferred embodiment includes compounds of formula I, wherein

N®-R ⁵ represents the remainder of the formula

and)

where R ³ is alkyl with 1 to 4 carbon atoms, preferably methyl and R 4 is hydrogen or alkyl with 1 to 4 carbon atoms,

b) _n 5

S where R ⁵ is alkyl with 1 to 4 carbon atoms, preferably methyl and R ⁵ and R ⁷ are hydrogen atoms or alkyl with 1 to 4 carbon atoms, ^C ' Λ® ₅

RN NR ⁵ where R is alkyl with 1 to 4 carbon atoms, preferably methyl and R is alkyl with 1 to 4 carbon atoms or phenylalkyl with 1 to 4 carbon atoms,

d)

ÑTÇ^rG

R° where R ⁵ is alkyl of 1 to 4 carbon atoms, preferably methyl, and R 6 is alkyl of 1 to 4 carbon atoms.

preferably methyl,

E)

RS'©

-C)

N—N _/ where R is alkyl of 1 to 4 carbon atoms, preferably methyl and R is alkyl of 1 to 4 carbon atoms, preferably methyl or

f) where R ⁵ is alkyl with 1 to 4 carbon atoms, preferably methyl.

Of the above-described embodiment of the present invention, those compounds where A is ^- ' ^CH 2>n ^- ' ⁿ 3 ^e 1 ^or ° 2» ³ are preferred, preferably those compounds where A is -Cl^- and where

8

a) R and R together form

or

1

b) R is a hydrogen atom and R is a hydrogen atom, CH ₃ CH ₂ ~,

^CH 3 ^Cii 3^ OH OH é- or ch ₃ -ch· ^CH 3 Q Particularly preferred are compounds where R is an atom hydrogen and R ¹ is OH i CH ₃ -CH-

especially those compounds that have the absolute configuration 5R, 6S, 8R.

The most preferred embodiment of the present invention includes compounds of general formula I, wherein

4- nr ⁵ represents the rest of the formula

b)

d)

F)

h)

j)

1)

n)

CH

CH, ch ₃ /v

N^N^CH ch ₃

Of the above-described embodiments, those compounds are preferred where A is ^-CH2 -n ^{and n} is 1 or 2, more preferred are those compounds where A is -(C2-)- and where

8

a) R and R together form boy ₂

or

b) R° is a hydrogen atom and R ¹ is a hydrogen atom, CH ₃ CH ₂ -,

CH.

CH.

Particularly preferred are those compounds wherein R ⁸ is or hydrogen and R 8 is

CH especially those compounds that have the absolute configuration 5R, 6S, 8R.

Specifically preferred compounds of the present invention are compounds of the formula

where R. is a hydrogen atom, an anionic charge or a common, easily cleavable carboxyl protecting group, whereby if R is a hydrogen atom or a protecting group, it may also be present in the form of an ion and where

-S-A

NR ⁵ is

a) _SCH ₂ -/ — CH,

b) -5ΟΗ ₂ ΟΗ ₂ -ΑΛ|-ΟΗ3

CH,

d) - SCH;

f) -sch ₂ -<( n-ch ₂ ch ₂ ch ₃ where

3.12

The HNMR ( ^D 2°) spectrum shows characteristic peaks at delta: 1.23 (3H, d, J=6, 4 Hz), (2H, q, J=1.4, 8.9 Hz), 3.39 (1H, q, J=2.7, 6.0 Hz), 4.07 to 4.68 (10H, m), 8.19 (1H, s);

q) ^- SCH'

CH,

CH, where

3.15

The HNMR ( _D2O ) spectrum shows characteristic peaks at delta: 1.23 (3H, d, J=6.4 Hz), (2H, q, J=3.7, 9.0 Hz), 3.37 (1H, q, J=2.6, 6.0 Hz), 3.95 to 4.65 (10h, m), 8.62 (1H, s);

r)

SCH _S / ^{N CH} 3

with)

CH,

t) φ

-sch ₂ --/ \

at)

COO .Θ ch ₂ i® sch ₂ -ZV ¹ n-ch ₃

CH, n-ch ₂ coo' ^SCH Z~\ /

N-N /

ch ₃ or

CH

in) ch ₃

The most preferred embodiment of the present invention includes compounds of general formula I, wherein represents

Of the above-described embodiment of the present invention, those compounds are preferred where A is -CH ₂ - _and where n is 1 or 2, more preferred are those where A is -CH ₂ - and where a) R and R together form a group of ₂ or

b) R ⁸ is

hydrogen atom a is a hydrogen atom, ch ₃ ch ₂ -,

CH_

CH„ OH 3-. I ?H-,

OH ch ₃ Ih- .

Particularly preferred are compounds where R is a hydrogen atom.

OH l

CH ₃ CHespecially those compounds that

Carbapenem derivatives of formula II have the absolute 5R configuration, of general formula I are prepared

6S, 8R.

from starting materials of general

R ⁸ H

H.-shore "15

I.S.A

COOR ² (II) which are reacted in an inert organic solvent with at least one equivalent (up to about 50% molar excess) of an alkylating agent of the formula wherein r 5 is as defined above and S' is a suitable leaving group such as a halogen atom (chlorine bromine or iodine, preferably iodine) or a sulfonic acid ester moiety such as mesylate, tosylate or triflate.

Examples of suitable non-reactive organic solvents are chloroform, methylene chloride, tetrahydrofuran, dioxane, acetone, dimethyl sulfoxide and dimethylformamide. The temperature for the alkylation reaction is not critical and temperatures in the range of about 0°C to 40°C are preferred. Preferably, the reaction step is carried out at room temperature.

ï) has a counterion X' (e.g. derived from the alkylating agent used) which can be replaced at this stage or at a later stage, e.g. after the cleavage step, by another ion, e.g. in a conventional manner by a more pharmaceutically acceptable ion. Alternatively, the ion can be removed during the deprotection reaction.

2' The step of cleaving the protecting group R from the carboxyl group of intermediate I is carried out by conventional methods such as solvolysis, chemical reduction or hydrogenation. If a protecting group such as p-nitrobenzyl, benzyl, benzhydryl or 2-naphthylmethyl is used, which can be removed by catalytic hydrogenation, intermediate X' in a suitable solvent such as a mixture of dioxane-water-ethanol, tetrahydrofuran-aqueous potassium hydrogen phosphate-isopropanol or the like is hydrogenated under atmospheric or slightly elevated pressure of 0.1 to 0.4 MPa of hydrogen over a hydrogenation catalyst such as palladium on carbon, palladium hydroxide, platinum oxide or the like, at a temperature of from 0 to 50 ° C for about 0.24 to 4 hours.

' oh, _in ,

If the R group is o-nitrobenzyl, then photolysis can be used to remove the protecting group. Protecting groups such as 2,2,2-trichloroethyl can be removed by mild reduction with zinc.

The allyl protecting group can be removed with a catalyst containing a mixture of a palladium compound and triphenylphosphine in an aprotic solvent such as tetrahydrofuran, diethyl ether or methylene chloride.

Similarly, other common carboxyl protecting groups can be removed by methods commonly known in the literature. Finally, as noted above, compounds of formula I where R is a physiologically hydrolyzable ester such as acetoxymethyl, phthalidyl, indanyl, pivaloyloxymethyl, methoxymethyl, etc., can be administered directly to the host without removal of the protecting groups, as these esters are hydrolyzable in vivo under physiological conditions.

5 15

It is understood that where an R, R, R or R substituent or heterocyclic ring is attached to a substituent A containing a functional group which may interfere with the intended course of the reaction, then this group may be protected with a conventional protecting group and then upon removal the desired functional group is regenerated.

Suitable protecting groups and procedures for introducing and removing such groups are commonly known in the literature.

In a variation of the above procedure, the carboxyl protecting group of intermediate II may be removed prior to the quaternization step. Thus, the carboxyl protecting group is removed as described above to yield the corresponding free carboxylic acid, and the free acid is then quaternized with an alkylating agent R -X' to provide the desired quaternized product of formula I.

If the intermediate IIa without a protecting group is quaternized, then water or a non-reactive organic solvent or a mixture thereof can be used as the solvent. Examples of suitable solvents are water, organic solvents such as chloroform, methylene chloride, tetrahydrofuran, dioxane, acetone, dimethylsulfoxide and dimethylformamide and water-organic solvent mixtures such as water-acetone or water-dimethylformamide.

The temperature for quaternization of intermediate IIa is not critical and temperatures ranging from about -40°C to about room temperature can conveniently be used. Preferably, the reaction is carried out at about 0°C.

If intermediate IIa is obtained as a carboxylic acid salt, it is necessary to add a strong acid such as toluenesulfonic acid to liberate the free carboxylic acid prior to quaternization.

This was found to greatly facilitate preferential quaternization at the nitrogen atom in the ring.

The above described variant is particularly useful if the carboxyl protecting group is more easily removed from the unquaternized intermediate XI than from the quaternized intermediate I. For example, in the preparation of the product of formula

R) (R) from the intermediate of the formula

removal of the allyl protecting group before quaternization leads to better yields of the desired final product.

The starting compounds of general formula J used for the process of the present invention can be prepared from compounds of general formula III.

(III) where R, R and R are as defined above and where ^R represents a conventional readily cleavable carboxyl protecting group. Compounds of formula III are disclosed, for example, in European Patent Application Publication No. 38,869 (compound 7) and in European Patent Application Publication No. 54,917 and can be prepared by the general methods described above.

The procedure for preparing compounds of formula I from starting materials of formula III can be summarized in the following reaction scheme:

*

5»»

L = regular leaving group

A variation of the above procedure is evident from the following reaction scheme

R® o1---1--= _d 15

HS-A(IV)

COOR ²

R®

R ¹⁵

With A

COOR ² removal of the protecting group

(III

If the above-described procedure is followed, the starting material of formula XII is reacted in an inert organic solvent such as methylene chloride, acetonitrile or dimethylformamide with an approximately equimolar amount of a reagent R°-L, such as p-toluenesulfonic anhydride, p-nitrobenzenesulfonic anhydride, 2,4,6-triisopropylbenzenesulfonic anhydride, methanesulfonic anhydride, trifluoromethanesulfonic anhydride, diphenylchlorophosphate, toluenesulfonyl chloride, p-bromobenzenesulfonyl chloride and the like, where L is an appropriate leaving group such as toluenesulfonyloxy, p-nitrobenzenesulfonyloxy, diphenoxyphosphinyloxy and other leaving groups which can be removed by conventional methods known from the literature.

The reaction for removing the leaving groups at position 2 of intermediate III is preferably carried out in the presence of a base such as diisopropylethylamine, triethylamine, 4-dimethylaminopyridine, etc. at a temperature of from about -20°C to +40°C, preferably at a temperature of about 0°C.

The leaving group L of intermediate IV may also be a halogen atom, and in this case this group is cleaved by reacting intermediate III with a halogenating agent such as O^PCl ₃ and (00) ^PB^, oxalyl chloride, etc. in a solvent such as dichloromethane, acetonitrile, tetrahydrofuran, etc., in the presence of a base such as diisopropylethylamine, triethylamine, 4-dimethylaminopyridine, etc.

Intermediate IV can optionally be isolated, but is preferably used in the next step without isolation or purification.

Intermediate IV is then converted to intermediate II by a conventional reaction. Thus, intermediate IV can be reacted with an approximately equimolar amount of a heteroarylalkyl mercaptan of the general formula

where A is a straight or branched alkylene chain of 1 to 6 carbon atoms and represents a mono-, bi- or polycyclic aromatic heterocyclic radical containing a quaternizable nitrogen in the ring, said ring being attached to A through a carbon atom. '.mhu, in an inert organic solvent such as dioxane, dimethylformamide, dimethylsulfoxide or acetonitrile and in the presence of a base such as diisopropylethylamine, triethylamine, sodium bicarbonate, potassium carbonate or 4-dimethylaminopyridine.

The temperature for the cleavage is not critical, but is preferably in the range of about -40° C. to 25° C. The reaction is most preferably carried out under cooling, e.g. at a temperature of about 0° C. to -10° C.

While the above-described procedure is suitable for preparing the compounds of the present invention, colleague Pierre Dextraze has discovered a novel procedure for preparing compounds of formula I. This alternative procedure, which is disclosed and claimed in a copending U.S. patent application filed on the same date as this application, is described below by way of example.

In an alternative process for preparing compounds of formula I, the intermediate of formula IV is

(IV) where R1 and ^R8 are as defined above, is a conventional easily removable carboxyl protecting group and L is a conventional leaving group such as toluenesulfonyloxy, ρ-nitrobenzenesulfonyloxy, diphenyloxyphosphinyloxy or a halogen atom is reacted with a thiol of formula VII

HS-A (VII) where A and are as defined above and X is an anion, in an inert solvent and in the presence of a base to form a carbapenem product of general formula I' _n 15

R° H

NR ³

2.....-N - \ ,/

About COOR ²

ï) where R ¹ , R ⁸ , R 8 , A, R ¹ - 2' and X have the meanings given above and optionally the carboxyl protecting group R is removed to obtain the corresponding deprotected compound of general formula I or a pharmaceutically acceptable salt thereof.

An alternative procedure uses an intermediate of the formula

(IV) which as mentioned above has been claimed for example in European Published Patent Application Nos. 38,869 and 54,917 and which can be prepared by the general methods described in that application.

L represents a common leaving group (defined as X in European Patent Application Publication No. 38,869, such as chlorine, bromine, iodine, benzenesulfonyloxy, p-toluenesulfonyloxy, p-nitrobenzenesulfonyloxy, methanesulfonyloxy, trifluoromethanesulfonyloxy, diphenoxyphosphinyloxy or di(trichloroethoxy)phosphinyloxy. A preferred leaving group is diphenoxyphosphinyloxy.

Intermediates of formula IV are generally formed in situ by the reaction of an intermediate of formula III

R ⁸ H _r 15

O coor ^2< (III) where R ¹ , R ⁸ , R ¹⁵ and R ² are as defined above, with a suitable acylating agent R°-L. A preferred intermediate IV, where L is a diphenoxyphosphinyloxy group, can be prepared by reacting ketoester III in an inert organic solvent such as methylene chloride, acetonitrile or dimethylformamide with an approximately equimolar amount of diphenylchlorophosphate in the presence of a base such as diisopropylethylamine, triethylamine, 4-dimethylaminopyridine and the like, at a temperature of from about -20°C to +40°C, preferably about 0°C. Intermediate IV can optionally be isolated, but is preferably used as a starting material for an alternative process without isolation or purification.

Carbapenem intermediate IV is reacted with a quaternary aminothiolein of the general formula

VII

has the meaning defined above and X is an anion. The reaction is carried out in an inert solvent such as acetonitrile, acetonitrile-dimethylformamide, tetrahydrofuran, tetrahydrofuran-water, acetonitrile-water or acetone in the presence of a base.

The type of base is not critical. Suitable bases are sodium hydroxide, diisopropylethylamine,

1,8-diazabicyclo(5.4.0)undec-7-ene, 1,5-diazabicyclo(4.3.0)non-5-ene and trialkylamines with 1 to 4 carbon atoms in each of the alkyls, such as triethylamine, tributylamine or tripropylamine.

The reaction of intermediate IV and thiol VII can be carried out over a wide temperature range, for example from -15°C to room temperature, but is preferably carried out at a temperature in the range of about -15°C to +15°C, most preferably around 0°C.

The carbapenem products prepared by reacting the quaternary amine thiol VII with the intermediate of general formula IV are linked to an anion (for example, (C^H^O) ₂ ?Ο ₂ Θ, Cl θ or an anion linked to a quaternary thiol), which at this stage can be replaced by another anion, for example one that is more pharmaceutically acceptable. This exchange is carried out in a conventional manner. Alternatively, the anion can be removed during a subsequent deprotecting step.

If the quaternized carbapenem compound and the anion form an insoluble product, then this product can be crystallized and collected pure by filtration.

To form the desired carbapenem product, the R ₂ protecting carboxyl group in compound I' can optionally be removed by conventional methods such as solvolysis, chemical reduction or hydrogenation. If a protecting group such as p-nitrobenzyl, benzyl, benzhydryl or 2-naphthylmethyl is used, which can be removed by catalytic hydrogenation, then. intermediate I* in a suitable solvent such as dioxane-water-ethanol, tetrahydrofuran-diethyl ether-buffer, tetrahydrofuran-aqueous potassium hydrogen phosphate-isopropanol, or the like. can be reacted under a hydrogen pressure of from 0.1 to 0.4 MPa 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 °C for 0.24 to 4 hours.

If the group is o-nitrobenzene, then photolysis can also be used to remove the protecting group. Protecting 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 comprising a mixture of a palladium compound and triphenylphosphine in a suitable aprotic solvent such as tetrahydrofuran, methylene chloride or diethyl ether.

Similarly, other common carboxyl protecting groups can be removed by methods commonly known in the art. Finally, as noted above, compounds of formula I wherein R is a physiologically hydrolyzable ester such as acetoxymethyl, ^phthalide , iudanyl, pivaloyloxymethyl, methoxymethyl, and the like can be applied directly to the host without removal of the protecting groups, since these esters are hydrolyzable in vivo under physiological conditions.

Thiol intermediates of formula VII can be prepared, for example, from the corresponding thiolacetate compounds of the general formula

where A has the meaning given above and

represents a mono-, bi- or polycyclic aromatic heterocyclic radical containing a quaternizable nitrogen atom in the ring, said ring being attached to A through a carbon atom in the ring. The thiolacetate compound is quaternized by reaction in an inert organic solvent such as diethyl ether, dichloromethane, methylene chloride, dioxane, benzene, xylene, toluene or mixtures thereof with a suitable alkylating agent of the general formula

X where R is as defined above and X is a conventional leaving group such as a halogen atom (chlorine, bromine or iodine, preferably iodine) or an ester moiety of a sulfonate such as mesylate, tosylate or triflate. The temperature for the alkylation reaction is not critical and temperatures in the range of about 0°C to 40°C are preferred.

Prior to reaction with carbapenem intermediate IV, the quaternized thiolacetate compound is subjected to acidic or basic hydrolysis to release the quaternary thiol intermediate VII. This hydrolysis is preferably carried out immediately prior to reaction with the compound of formula IV so as to minimize decomposition of the relatively unstable quaternary thiol VII.

By appropriate selection of solvents, the reaction of intermediate III to final product I can be carried out without isolation of the various intermediates, i.e., by the so-called "one-flask" process.

An example of such a procedure is shown below in Example 22.

As with other beta-lactam antibiotics, compounds of formula I can be converted by known methods into pharmaceutically acceptable salts which, for the purposes of the present invention, are substantially equivalent to the compounds not in salt form.

For example, a compound of formula I, wherein R is an anionic charge, can be dissolved in a suitable inert solvent and then an equivalent of a pharmaceutically acceptable acid added. The desired acid addition salt can be isolated by conventional methods, for example, solvent precipitation, lyophilization, and the like.

Where other basic or acidic functional groups are present in the compounds of formula I, pharmaceutically acceptable base addition salts and acid addition salts can be prepared analogously by known methods.

It is understood that certain products falling within the scope of formula I may occur in the form of optical isomers as well as in the form of epimeric mixtures. All optical isomers and epimeric mixtures are included within the scope of the present invention.

For example, if the substituent at position 6 is hydroxyethyl, then the substituent may be in either the R or S configuration and the resulting isomers, as well as epimeric mixtures, are within the scope of the present invention.

A compound of formula 1, wherein R is a hydrogen atom or an anionic charge, or a pharmaceutically acceptable salt thereof.

a suitable salt, can also be converted by conventional methods into the corresponding compound where R is a physiologically hydrolysable ester group or a compound of formula I where R is a conventional carboxyl protecting group, can be converted into the corresponding compound where R is a hydrogen atom, an anionic charge or a physiologically hydrolysable ester group or a pharmaceutically acceptable salt thereof.

The novel carbapenem derivatives of the general formula I, where R ² is a hydrogen atom, an anionic charge or its physiologically hydrolyzable carboxyl protecting group, or their pharmaceutically acceptable salts, are effective antibiotics against various gram-positive and gram-negative bacteria and can be used, for example, as feed additives to enhance growth, as food preservatives, as bactericides for industrial applications, for example for water-soluble dyes and circulating waters in paper mills, where they inhibit the growth of harmful bacteria, and as disinfectants for the decomposition and inhibition of the growth of harmful bacteria in medical and dental instruments.

However, they are particularly useful in treating infectious diseases in humans and other animals caused by gram-positive or gram-negative bacteria.

The pharmaceutically active compounds of the present invention may be used alone or may be formulated into pharmaceutical compositions containing the active carbapenem components in a pharmaceutically acceptable carrier or diluent.

The compounds can be administered in various ways, with oral, topical, or parenteral (e.g., intravenous or intramuscular injection) being the main routes of administration. The pharmaceutical compositions may be solid, for example, in the form of tablets, powders, capsules, etc., or liquid, such as solutions, suspensions, or emulsions.

Compositions for injection, a preferred route of administration, may be prepared in unit dosage form in ampoules or in multi-dose containers and may contain auxiliary agents such as suspending, stabilizing and dispersing agents. The compositions may be in a form ready for use for administration or in powder form for reconstitution at a convenient time by the addition of a suitable carrier such as sterile water.

The dosage to be administered will depend in particular on the compound employed, the particular formulation of the mixture, the route of administration, the type and condition of the patient and the target site, and the organism to be treated. The selection of the particular preferred dosage and route of administration is then left to the experience of the physician. In general, however, the compounds are administered parenterally or orally to a mammal in an amount of from about 5 to 200 mg/kg/day.

The application is generally done in divided doses, that is, three to four times a day.

To illustrate the effective broad spectrum antibacterial activity of the carbapenems of the invention, both in vitro and in vivo, and the low toxicity of the compounds, the biological data presented below are compared with preferred carbapenem compounds of the present invention.

In vitro activity

Samples of the carbapenem compounds prepared in Examples 1 and 2 after dissolution in water and dilution with nutrient medium show the following minimum inhibitory concentrations (MIC) in /ug/ml against the indicated microorganisms. The test was determined by incubation overnight at 37 °C- dilution in a test tube. N-formimidoylthiermamycin was used as a reference compound,

In vitro antibacterial activity of the carbapenem derivative according to Example 1

Organism MIC (µg/ml)

New compound N-formimidoylthienamycin

p. pneumoniae A-9585 0.25 0.004 p. Pyogenes A-9604 0.06 0.001 p. aureus A-9537 0.13 0.004 p. aureus + 50% serum A-9537 0.03 0.016 p. aureus (Pen-res.) 1-9606 0.06 0.008 p. aureus (Meth-res.) A-15097 4 0.5 p. faecalis A20688 0.5 0.5 E. coli (10 ⁴ dilution) A15119 0.06 0.016 E. coli (10~ ³ ) A15119 - 0.03 E. coli (ΙΟ ^-2 ) A15119 - 0.06' E. coli (10~ ⁴ ) A20341-1 0.03 0.03 E. coli (10 ^-3 ) A20341-1 - 0.03 E. coli dO' ² ) A20341-1 - 0.13 K. pneumoniae A-9664 0.25 0.13 K. pneumoniae A20468 0.25 0.06

table continuation

Organism MIC (μg/ml) New compound N-formimidoylthienamycin P. mirabilis A-9900 0.13 0.06 P. vulgaris A21559 0.13 0.03 P. morganii A15153 0.13 0.13 P. rettgeri A22424 0.25 0.25 S. marcescens A20019 0.13 0.03 E. cloacae A-9596 0.13 0.06 E. cloacae A-9656 0.13 0.06 P. aeruginosa A-9843A 4 1 P. aeruginosa A21213 1 0.25 H. influenzae A-9833 16 16 H. influenzae A20178 32 32 H. influenzae A21522 8 32 B. fragilis A22862 0.03 0.016 B. fragilis A22053 0.03 0.06 B. fragilis 122696 0.25 0.13 B. fragilis A22863 0.03 1

In vitro antibacterial activity of the carbapenem derivative according to Example 2

Organism MIC (µg/ml)

New compound N-formimidoylthienamycin

p. pneumoniae A-9585 0.001 0.002 p. pyogenes A-9604 0.001 0.002 p. aureus A-9537 0.004 0.004 p. aureus A-9537 + 50% serum 0.016 0.016 p. aureus A-9 6 0 6 (Pen-re s.) 0.008 0.008 p. aureus (Meth-res.) A15097 8 4 p. faecalis A20688 0.25 0.5 E. coli A15119 0.016 0.016 E. coli A2034L-1 0.016 0.03 K. pneumoniae A-9664 0.06 0.06 P. mirabilis A-9900 0.03 0.06 P. vulgaris A21559 0.016 0.03 P. morganii A15153 0.06 0.13 P. rettgeri A22424 0.13 0.13 p. marcescens A20019 0.03 0.03 E. cloacae A9659 0.13 0.06 E. cloacae A9656 0.25 0.06 P. aeruginosa A9843A 8 1 P. aeruginosa A21213 2 0.25 In vivo activity Therapeutic effect in vivo compounds according to example 1 and N^formimidoylthienamycin

after intramuscular administration to mice experimentally infected with various organisms is shown in the following table. The Ρϋ^θ values (dose in mg/kg required for 50% protection of infected mice) are as follows:

Protective effect of intramuscular treatment of infected mice

Organism Infection (number of organisms) PDj-0 (treatment (mg/kg) Compound according to Example 1 N-formimidoyl- thienamycin P. , mirabilis A-9900 3.6 x 10 ⁶ 3.3 ^3X / ^15x P. . aeruginosa A-9843a 5.5 x 10 ⁴ 0.3 ^0.5x P. . aeruginosa A-20481 5.4 x 10 ⁴ 0.63 ^0.4X P, , aeruginosa A-20599 1.4 x 10 ⁵ 0.7 ^0.18x p. golden A-9606 6.6 x 10~ ⁸ 0.09 ^0.07x p. faecalis A-20688 2.3 x 10 ⁸ 3.3 ^2.8X E. coli A-15119 6.2 x 10 ⁶ 0.6 ^2.5X K. . pneumoniae A-9964 5.1 x 10® 2.5 ^2.2X

Historical data Treatment sequence: With the exception of E. coli A15119 and K. pneumoniae A9964, mice were treated i.m. with drug at 0 and 2 hours post infection. For E. coli and K. pneumoniae, treatment was at 1 and 3.5 hours post infection. 5 mice per dose were used in each test.

Toxicity

The toxicity of the compound of Example 1 was determined by intracranial administration to mice and the results are summarized in the table below:

Toxicity for intracranial administration to mice

Compound

Highest dose (mg/kg) x _T Without clinical ^signs of toxicity (mg/kg) compound of Example 1

N-formimidoylthienamycin

5

2 ^5 ^x average 25/mouse/compound

Blood levels in mice after intramuscular administration

The blood levels and half-life of the compound of Example 10 mg/kg mouse are shown in the following table:

after intramuscular administration

Compound 10 20 Level in 30 45 blood 60 (^ug/ml) 90 ^x tl/2 (min) ^XX AUC (^ug.h/ml) compound according to example 1 14 10.8 6.8 2.6 0.8 PLN 0.6 10 6.3 N-formimidoylthienamycin 12.6 9.9 7.3 2.6 0.7 < 0.3 9 6

Compounds are dissolved in 0.1M phosphate buffer pH 7. Values are from one test, 4 mice per compound were used. ^X t1/2 - half-life in minutes ^xx AUC - refers to the area under the curve.

Urine excretion

The urinary excretion of the compound of Example 1 after intramuscular administration (20 mg/kg) to mice is evident from the following table:

Urinary excretion after intramuscular administration of 20 mg/kg to mice

Compound Percentage of excluded dose 0-3 3-6 hours after 6-24 applications 0-24 compound according to example 1 26.1 0.5 0.1 26.7 + 6.7 N-formimidoylthienamycin 12.1 0.1 <0.1 12.2 + 3.6

Compounds are dissolved in 0.1M phosphate buffer pH 7. Values are from one test, 4 mice per compound were used.

In vitro activity

The carbapenem compound samples listed below (identified by example numbers) for dissolution in water and dilution with nutrient medium show the following minimum inhibitory concentrations (MIC) in µg/ml against the indicated microorganisms. The test was performed by overnight incubation at 37°C with dilution in a test tube. In the following tables, N-formimidoylthienamycin is used as a reference compound.

Organism MIC (µg/ml)

ex. 3 ex. 4 ex. 5 ex. 6 ex. 7 MK 0787 p. , pneumoniae 1-9585 0.002 0.002 0.004 0.004 0.004 0.002 p. pyogenes A-9604 0.002 0.001 0.004 0.004 0.004 0.002 p. , faecalis A20688 0.5 0.5 0.25 0.5 0.13 0.5 p. . aureus A-9537 0.016 0.008 0.004 0.03 0.008 0.004 5. . aureus + 50% serum 1-9537 0.016 0.03 0.016 0.06 0.03 0.016 p. golden A-9606 (Pen-res) 0.016 0.016 0.008 0.03 0.016 0.008 P. golden A15097 4 4 8 4 2 4

(Meth-res)

E. coli A15119 0.03 0.016 0.016 0.06 0.004 0.016 E. coli A20341-1 0.016 0.016 0.016 0.06 0.004 0.03 K. pneumoniae A-9664 0.06 0.03 0.03 0.13 0.016 0.06 K. pneumoniae 120468 0.06 0.03 0.06 0.25 0.016 0.13 E. cloacae A-9659 0.13 0.03 0.06 0.25 0.016 0.06 E. cloacae A-9656 0.13 0.06 0.13 0.25 0.016 0.06 P. mirabilis A-9900 0.13 0.06 0.03 0.025 0.016 0.06 P. common A-21559 0.016 0.016 0.016 0.06 0.008 0.03 M. morgania A15153 0.13 0.016 0.06 0.13 0.016 0.13 P. rettgeri A22424 0.25 0.13 0.13 0.25 0.06 0.13 P. marcescens A20019 0.016 0.03 0.06 0.13 0.008 0.03 P. aeruginosa A-9843a 2 2 4 8 1 0.25 P. aeruginosa A21213 0.5 0.13 2 1 0.25 0.25 H. influenza A-9833 2 2 4 >32 >32 16 H. influenza A21518 2 2 4 732 >32 16 B. fragilis A22862 0.25 0.06 0.03 0.03 0.016 0.06 B. fragilis A22696 0.5 0.5 0.25 0.25 0.25 0.13

^x N-formimidoylthienamycin

Organism ex. 8 MIC (µg/ml) ex. 9 ex. 10 ex. 11 MK 0787 P. pneumoniae A-9585 0.002 0.001 0.001 0.002 0.002 P. pyogenes A-9604 0.002 0.001 0.001 0.002 0.0Q2 P. faecalis A20688 0.5 0.13 0.25 0.5 0.25 P. golden A-9537 0.008 0.004 0.008 0.004 0?002 P. golden A-9537 0.03 0.008 0.06 0.016 0.016

(50% serum)

P. golden A-9606 0.03 0.008 0.008 0.016 0.008 (Pen-res) p. golden A15097 - - - (Meth-res) E. coli A15119 0.016 0.008 0.016 0.016 0.016 E. coli A20341-1 0.03 0.004 0.008 0.03 0.016 K. pneumoniae A-9664 0.03 0.016 0.06 0.03 0.03 K. pneumoniae A20468 0.13 0.03 0.13 0.13 0.13 E. cloacae A-9659 0.13 0.03 0.013 0.06 0.13 E. cloacae A-9656 0.06 0.03 0.13 0.13 0.06 P. mirabilis A-9900 0.13 0.016 0.13 0.03 0.03 P. common A21559 0.03 0.008 0.016 0.03 0.016 M. morgania A15153 0.13 0.03 0.13 0.06 0.06 P. rettgeri A22424 0.13 0.06 0.13 0.13 0.13 P. marcescens A20019 0.06 • 0.016 0.06 0.06 0.03 P. aeruginosa A-9843A 1 2 32 0.5 1 P. aeruginosa A21213 0.25 0.13 2' 0.13 0.13 ^X N- -formimidoylthienamycin

Organism pr. 12 MIC yug/ml) ex. 13 ex. 14 MK 0787 ^X P. , pneumoniae A-9585 0.002 0.0005 0.0005 0.002 P. pyogenes A-9604 0.004 0.0005 0.0005 0.002 P. faecalis A20688 0.5 0.13 0.13 0.25 P. . aureus A-9537 0.008 0.008 0.008 0.002 p. golden A-9537 0.016 0.016 0.03 0.008 + 50% serum p. golden A-9606 0.03 0.008 0.016 0.008 (Pen-res) p. golden A15097 - - - - (Meth-res) E. coli A15119 0.016 0.004 '0.008 0.016 E. coli A20341-1 0.008 0.008 0.008 0.016 K. pneumoniae A-9664 0.03 0.03 0.03 0.03 K. pneumoniae A20468 0.06 0.13 0.03 0.06 E. cloacae A-9659 0.06 0.06 0.03 0.06 E. cloacae A-9656 0.03 0.03 0.03 0.06 P. mirabilis A-9900 0.03 0.016 0.016 0.016 P. common A21559 0.016 0.008 0.016 0.016 M. morgania A15153 0.06 0.016 0.03 0.06 P. rettgeri A22424 0.13 0.25 0.06 0.13 P. marcescens A20019 0.03 0.016 0.016 0.03 P. aeruginosa A-9843A 16 32 8 1 P. aeruginosa A21213 2 2 0.5 0.13

^X N-formimidoylthienamycin

In vivo activity

The therapeutic effect of certain compounds of the present invention and N-formimidoylthienamycin (MK 0787) after intramuscular administration to mice experimentally infected with various organisms is shown below.

The PO ₅ θ values (dose in mg/kg) that provide 50% protection of infected mice are given.

Protective effect of intramuscular treatment of infected mice

PDjg/treatment (mg/kg)

ex. 6 ex. 8 ex. 9 ex. 12 ex. 14 MK 0787 p. golden A9606 - 0.21 - 0.89 0.07 0.07 E. coli A15119 - 0.86 1.2 - - 3 K. pneumoniae A9664 - 1.8 1.8 - - 3 P. mirabilis A9900 - 1.4 7.1 - - 9 P, aeruginosa A9843A 0.4 0.19 0.19 1.8 0.45 1 P, aeruginosa A24081 - 0.33 0.19 - - 0.4

ex. 3 ex. 4 ex. 7 MK 0787 p. golden A9606 0.07 0.1 0.2 0.07 E. coli A15119 1 0.4 0.2 3 K. pneumoniae A9664 3 3 1 3 P. mirabilis A9900 2 4 2.4 9' P. aeruginosa A9843A 0.5 0.2 0.2 0/5 P. aeruginosa A24081 0.8 0.2 0.1 0.4 ex. 5 MK 0 7 8 7 P. golden 0.2 0.07 E. coli 4 2.2 K. pneumoniae 3 2.3 P. mirabilis 10 9 P. aeruginosa A9843A 1.6 0.5

Blood levels in mice after intramuscular administration

The blood levels and half-lives of certain compounds of the present invention following intramolecular administration of 20 mg/kg to mice are shown below.

Compound Cmax (μg/ml) * ^T1 /2 (min) ^XX AUC (log.h/ml) example 1 14 10 6.3 example 2 13.9 9 5.3 example 3 14.5 10 • 6.9 example 4 15.5. 11 7.7 example 5 - - example 6 17.7 9 8.5 example 7 19.2 11 11.8 example 8 18.8 11 10.5 example 9 16.7 12 oh , 5

table continuation

Compound Cmax (/µg/ml) x T _1/2 (min) ^XX AUC (µg.h/ml) example 10 20.1 11 9.5 example 11 14.9 11 7.4 example 13 14.8 11 6.4 example 14 15.8 13 7.6

Compounds are dissolved in 0.1M phosphate buffer, pH 7. Values are from one test, one mouse per compound.

x ~ P° ^{time in} minutes ^XX AUC - area under the blood concentration curve over a period of time

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

Example 1

Preparation of the inner salt of 1-methyl-4-[2-carboxy-6alpha-[1(E)-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-3-thiomethyl]pyridinium hydroxide

A solution of 673 g (1.86 mmol) of p-nitrobenzyl 6alpha-[1-(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate in 10 ml of acetonitrile is cooled to -10 °C under a nitrogen atmosphere.

%

A solution of 245 mg (1.90 mmol) of diisopropylethylamine in 1 ml of acetonitrile is added, and then 510 g (1.90 mmol) of diphenylchlorophosphate in 1 ml of acetonitrile is added dropwise over two minutes.

The resulting solution was stirred for 15 minutes at -10 °C to give p-nitrobenzyl-3-(diphenylphosphoryloxy)-6alpha-[1-(R(-hydroxyethyl)-7-oxo-1-azabicyclo[3.2.0]-hept-2-ene-2-carboxylate.

To this solution was added a solution of 245 mg (1.90 mmol) of diisopropylethylamine in 0.5 mL of acetonitrile followed by a solution of 270 mg (2.16 mmol) of 4-mercaptomethylpyridine in 0.5 mL of acetonitrile. The reaction mixture was stirred at -10 °C for 60 min and the resulting white precipitate was filtered off and washed with 5 mL of ice-cold acetonitrile to give 660 mg (76% yield) of compound 2 as white crystals, mp 145 °C.

NMR (DMSO-d6) delta:

1.20 (3H, d, J = 6.0 Hz), 3.2 to 3.4 (3H, m), 3.7 to 4.25 (2H, s), 5.05 (1H, d, J = 4.0 Hz), 5.25 (1H, d, 5.48 (1H, d, J = 14.0 Hz), 7.40 (2H, d, J = 5.5 Hz), J = 8.5 Hz), 8.23 (2H, d, J = 8.5 Hz) and 8.58 )2H, d,

4.1 (2H, m),

J = 14.0 Hz), 7.70 (2H, d,

J = 5.5 Hz),

IC (KBr) gamma max: 3,400,

790,

695 and 1,600 cm

Analysis: calculated for ^C 22 ^H 21 ^N 3°6 ^S

58.01% C, 4.56% H, 9.23 found 57.74% C, 4.56% H, 9.58% N, 7.04% S. % N, 7.21% S.

To a solution of 660 mg (1.41 mmol) of intermediate 2 in 140 mL of acetone was added 5 mL of methyl iodide. The reaction solution was stirred for 8 h at 25 °C. The solvent was evaporated in vacuo and the resulting light yellow solid was triturated with diethyl ether to give 779 mg (90% yield) of compound 3 as a white amorphous solid, mp 130 °C (decomposition).

NMR (DMSO-d6) delta: 1.15 (3H, d, J = 6.0 Hz), 3.2 to 3.4 (3H, m), 3.7 to 4.1 (2H, m),

4.25 (3H, s), 4.30 (2H, s), 5.25 (1H, d, J = 14.0 Hz), 5.50 (1H, d, J = 14.0 Hz), 7.70 (2H, d, J = 9.0 Hz), 8.10 (2H, d, J = 7.0 Hz)

8.25 (2H, d, J· = 9.0 Hz) and 8.90 (2H, d, J = 7.0 Hz).

Infrared (KBr) gamma: 3,400, 1,770, 1,690 and 1,640 cm ^-1 .

max ·

Analysis for ^C 23 ^H 24 ^N 3°6 ^SI ' ^H 2° calculated 44.39% C, 4.22% H, 6.82% N, 5.20% S, found 44.66% C, 4.01% H, 6.84% N, 5.64% S.

To a solution of 779 mg (1.27 mmol) of compound 3 in a mixture of tetrahydrofuran, ether (80 ml-80 ml-100 ml) was added 140 mg (1.4 mmol) of hydrogen carbonate (0.7 mmol) of dibasic potassium phosphate.

water and diethyl potassium and 125 mg

Then 700 mg of 10% palladium on carbon were added and the mixture was hydrogenated for 45 minutes at 276 kPa on a Paar shaker. The mixture was filtered and the catalyst was then washed with water (2 x 10 ml). The combined filtrate and washings were extracted with diethyl ether (150 ml) and then lyophilized to give a brown powder.

This crude material was purified on a C^g BONDAPAK reversed phase column (30 g) Waters Associates) and eluted with water at 55.2 kPa. Each fraction (20 ml) was separated by high pressure liquid chromatography and the fractions with ultraviolet absorption at lambda _max = 300 nm were combined and lyophilized to give 135 mg (32% yield) of compound 4 as a pale yellow solid.

NMF t ( _D2O ) delta: 1.25 (3H, d, J = 6.0 and 2.5 Hz), 3.9 to 4, d, J = 6.0 Hz), 8.72 ID (KBr) ^gamma max : 3 400, 1,755, 1,640 UV lambda max ^(H 2' 0) : 296 nm (epsilon = to say harmony 2

Hz), 2.7 to 3.2 (2H, m), 3.40 (1H, q, J = 9.0 (2H, m), 4.40 (3H, s), 4.72 (2H, s), 8.10 (2H,

(2H, d, J = 6.0 Hz). and 1,590 -1 cm 7,782), 258 nm (epsilon = 6,913)

OH

S· ©

N-CH

OH

_CO2 pNB

_CO2 PNB

A suspension of carbapenem 3 (890 mg, 1.85 mmol) and 7 mL of iodomethane in 200 mL of anhydrous acetone and 12 mL of methylene chloride was stirred for 24 h at 25 °C. After 18 h, the reaction mixture became a clear solution. The solvent was removed under reduced pressure and the residue was then washed with diethyl ether to give 920 mg (1.48 mmol) (79.8%) of compound 4 as a foamy solid.

ID (KBr technique) 1,765, 1,690 cm

NMR (DMSO) delta: 1.3 (3H, d, J - 3.0 Hz), 3.1 to 3.7 (7H, m), 4.1 (3H, m), 4.3 (3H, s), 5.38 (2H, q, J = 7.0 Hz), 8.1 (2H, d, J = 3.0 Hz), 8.9 (2H, d, J = = 3.0 Hz), 7.6 (2H, d, J = 4.0 Hz), 8.2 (2H, d, J = 4.0 Hz).

B.

CHq

CO ₂ pNB (Η) γ

To carbapenem 4 (920 mg, 1.47 mmol) dissolved in 90 mL tetrahydrofuran, 90 mL diethyl ether, and 90 mL water was added 265 mg (1.51 mmol) dibasic potassium phosphate,

190 mg (1.9 mmol) of potassium bicarbonate and 800 mg of 10% palladium on carbon. The reaction mixture is hydrogenated for 1 hour at 310 kPa. The catalyst is filtered off through celite and the filtrate is washed with diethyl ether (3 x 25 ml).

The aqueous layer was lyophilized to give a brownish solid which was purified twice by chromatography on a 12 g C12BONDAPAK reversed phase column (Waters Associates) ( _H2O ) to give 55 mg of compound 5.

IR (KBr-teohnika): 1,750, 1,640 cm ³ .

NMR (D ₂ O) delta: 1.30 (3H, d, J = 3.0 Hz), 3.0 - 3.5 (7H, m), 4.3 (3H, s), 4.0 to 4.5 (3H, jn), 7.90 (2H, d), 8.70 (2H, d).

OH

ch ₃

Example 3

Preparation of 3-(N-methylpyridin-3-yl-methylthio)-6alpha-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

CH ₃ I

OH

Pd/C,H ₂

CH,

COjpNB

OH

CH,

WHAT,

1'

3-(N-Methylpyridin-3-yl-methanethio)-6alpha-Q-(R)-hydroxyethyl 3-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

To a solution of 730 mg (1.56 mmol) of compound 19 in 120 mL of acetone was added 5 mL of methyl iodide and the reaction mixture was stirred at room temperature for 18 h. The precipitate was filtered off and washed with acetone (10 mL) to give 940 mg (100% yield) of quaternized pyridine 20 as a light yellow powder.

NMR (DMSO-d6) delta: 1.25 (3H, d, J = 5.8 Hz), 3.6 to 4.3 (5H, ni) , 4.20 (3H, s) , 4.25 (2H, s), 5.25 (1H, d, J = 7.2 Hz), 5.40 <2H, ABq, J = 12, 16 Hz) and 7.6 to 9.2 (9H, m).

IC (KBR technique) lambda _max : 3,300, 1,765 and 1,690 cm” ¹ .

Analysis for ^C 23 ^H 24 ^N 3°6 ^S 1 ¹ 1 calculated 46.24%C, 4.05%H, 7.03%N, 5.37%S, found 45.82%C, 4.11%H, 6.87%N, 6.10%S.

To a solution of 933 mg (1.6 mmol) of compound 20 in 90 mL of tetrahydrofuran and 90 mL of ether was added 200 mg of KHCO^ and 249 mg of ^HPO^ in 90 mL of water, followed by the addition of 1.0 g of palladium on carbon.

The mixture is hydrogenated for 45 minutes at 310 kPc on a Paar shaker.

The reaction mixture was filtered through celite, the catalyst washed with water (2 x 10 ml). The combined filtrate and washings were extracted with ether (2 x 100 ml) and lyophilized to give a yellow solid which was purified on a C^g BONADAPAK (Waters Associates) reversed phase column (8 g), eluting with 5% acetonitrile in water at 55 kPa. Each 15 ml fraction was assayed by high pressure liquid chromatography and the fractions with ultraviolet absorption at lambda _max 300 nm were pooled and lyophilized to give 230 mg (43% yield) of the title compound as pale yellow crystals, mp 130 °C (dec).

NMR (D _? O) delta: 1.25 (3H, d, J = 7.0 Hz), 3.12 (2H, d.d., J = 7.9 Hz, 1.6Hz), 3.42 (1H, q, J =7.2 Hz), 1.6 Hz), 3 .9 to 4.6 (3H, m) , 4.25 (2H, s), 4.42 (3H, s) and 8.0 to 9.0 (4H, m). iC (technology KBr) gamma : max 3,400, 1,750 and 1,580 -1 cm in lambda max ( _H2O ): 298 nm (epsilon = 8,058) .

Analysis for ^c igH^gN ₂ O^S^.2H ₂ O calculated 51.87% C, 5.44% H, 7.56% N, found 51.95% C, 5.66% H, 7.56% N.

Example 4

Preparation of 3-(N-methylpyridin-2-ylmethanethio)-6alpha-[1-(R)hydroxyethyl-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

OH

_CO2 pNB

3-(N-Methylpyridin-2-ylineethanethio)-6alpha-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

To a solution of 650 mg (1.39 mmol) of compound 22 in 100 mL of acetone was added 4 mL of methyl iodide. The reaction mixture was stirred for 3 days at room temperature. The precipitate was filtered off and washed with acetone (10 mL) to give 500 mg (60% yield) of quaternized pyridine 2 as a pale yellow solid.

NMR 1DMSO-D6) delta: 1.26 (3H, d, J = 7.0 Hz), 3.9 to 4.2 (2H, m), 4.4 (3H, s), 4.78 (2H, s), 5.2 (1H, d, J = 3.8 Hz), 5.50 (2H, ABq, J = 14 Hz), and 7.8 to 9.4 (8H, m) .

IR (KBr-technique) gamma 3,400, 1,765 and 1,690 cm \ max

Analysis for ^C 23 ^H 24 ^N 3°6 ^S l' ⁱ 'l calculated 46.24% C, 4.05% H, 7.03% N, 5.37% S, found 45.62% C, 4.27% H, 6.80% N, 5.30% S.

To a solution of 1.0 g (1.167 mmol) of compound 23 in 90 ml of tetrahydrofuran and 90 ml of ether was added 215 mg (2.15 mmol) of KHCO^ and 374 g (2.1 mmol) of K^HPQ^ in 90 ml of water, followed by the addition of 1.0 g of 10% palladium on carbon. The mixture was hydrogenated at 310 kPa for 45 minutes on a Paar shaker.

The reaction mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL).

The combined filtrate and washings were extracted with ether (2 x 100 ml) and lyophilized to give a yellow solid which was purified on a BONDAPAK (Waters Associates) reversed phase column (10 g), eluting with 5% acetonitrile in water at 55 kPa. Each 15 ml fraction was assayed by high pressure liquid chromatography and the fractions with an ultraviolet absorption at lambda _max 300 nm were pooled and lyophilized to give 390 mg (44% yield) of the title compound. Recrystallization of this material from water-acetone-ethanol gave fine needles, mp 194-196 °C (dec.).

NMR (D ₂ O) delta: 1.30 (3H, d, J = 6.2 Hz), 3.2 (2H, q, J = 9.0 Hz, 3.6 Hz), 3.46 (1H, q,

J = 6.0 Hz, 2.7 Hz), 4.1 to 4.6 (3H, m), 4.60 (3H, s), 7.9 to 8.9 (4H, m).

TČ (KBr) 9 ^ama max ^: 3,400, 1,7555 and 1,590 cm.

UV lambda (H~O): 292 nm (epsilon = 8092). max of *

Analysis for ^c i6 ^H 28 ^N 2°4 ^S l' ^2H 2° calculated 51.87% C, 5.44% H, 7.56% N, found 51.37% C, 5.69% H, 7.37% N.

Example5 &

Preparation of 3-(N-methylpyridin-2-ylethanethio)-6alpha-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

_CO2 pNB

-£1-(R)-Hydroxyethyl 3-(N-methylpyridin-2-ylethanethio)-6alpha hept-2-ene-2-carboxylate

7-oxo-1-azabicyclo(3.2.0) To a suspension of 800 mg (1.7 mmol) of compound 53 in 50 mL of acetone was added 5 mL of methyl iodide. The reaction mixture was stirred for 48 h at room temperature. The precipitate was filtered off and washed with acetonitrile (15 mL) to give 810 mg (76% yield) of quaternized pyridine 55 as a pale yellow powder.

NMR (DMSO-d6) delta: 1.20 (3H, d, J = 5.6 Hz), 3.2 to 4.3 (9H, m), 4.20 (3H, s), 5.26 and 5.55 (1H each, ABq, J = 15 Hz) and 7.8 to 9.2 (8H, m).

ID (Kbr technique) 9 ^ama max ^; 3,400, 1,770 and 1,690 cm

To a solution of 790 mg (1.27 mmol) of compound 55 in 100 mL of tetrahydrofuran and 100 mL of ether was added 100 mL of pH 7.0 buffer, followed by the addition of 1.0 g of 10% palladium on carbon. The mixture was hydrogenated at 276 kPa for 40 minutes on a Paar shaker.

The reaction mixture was filtered through celite and the catalyst washed with water (2 x 10 ml). The combined filtrate and washings were extracted with ether (3 x 100 ml) and lyophilized to give a yellow powder which was purified on a BONDAPAK (Waters Associates) column (30 g), eluting with

10% acetonitrile in water at a pressure of 55 kPa.

Each 15 ml fraction was assayed by high-performance liquid chromatography and the fractions having ultraviolet absorption at lambda 300 nm were pooled and lyophilized to give 65 mg (15% yield) of the product as a yellow powder.

NMR ( _D20 ) delta: 1.25 (3H, d, J = 6 4.32 (3H, s) and 7m8 ID (KBr technique) gamma: max 3,400, 1 UV lambda (H _o 0): 300 nm max 2 (epsilon Example 6

Hz), 3.1 to 3.6 (7H, m), 4.0 to 4.3 (2H, m), to 8.9 (411, ra).

750, 1590 cm ¹ .

= 8 108).

Preparation of 3-(1-propylpyridin-4-yl methanethio-6a.lfa-[jL-(E)hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

acetone

Nah

-{1-allylpyridin-4-ylmethanethio)-6alpha-[1-(R)-hydroxyethyl-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-3-2-carboxylate

To a solution of 900 mg (2.13 mmol) of compound 51 in 150 mL of acetone were added 2 mL of allyl bromide and 380 mg of sodium iodide. The reaction mixture was stirred at room temperature for 48 h and the solvent was removed in vacuo to give a yellow solid.

This material was suspended in 120 ml of acetonitrile, and ^the reaction mixture was concentrated to give 1.0 g (87% yield) of the title compound as a yellow solid.

NMR (CD CH) delta: 1.20 (3H, d, J = 6.2 Hz), 3.0 to 3.4 (4H, m), 4.0 to 4.4 (4H, m), 5.1 to 5.6 (4H, m), and 7.4 to 7.9 (8H, m).

IR (KBr technique) gamma: 3,400, 1,770 and 1,690 cm

Analysis for ^C 25 ^řI 26 ^N 3°6 ^S l' ^I *l calculated 48.16% C, 4.21% H, 6.74% N, 5.15% G, found 48.55% C, 4.46% H, 6.69% N, 5.15% S.

24719

3-(1-propylpyridin-4-ylmethanethio)-6alpha-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

To a solution of 1.27 g (2.15 mmol) of compound 52 in 100 ml of tetrahydrofuran and 100 ml of ether was added 100 ml of buffer pH 7.0 and then 1.0 g of 10% palladium on carbon was added. The mixture was hydrogenated at 276 kPa for 40 minutes on a Paar shaker.

The reaction mixture was filtered through celite and the catalyst was washed with water (2 x 10 ml) and lyophilized to give a yellow powder which was purified on a _C1g BONDAPAK (Waters Associates) column (40 g), eluting with 10% acetonitrile in water at 55 kPa.

Each 15 ml fraction was tested by high-performance liquid chromatography and the fractions with ultraviolet absorption at lambda _max 300 nm were pooled and lyophilized to give 48 g (6% yield) of the title compound as a yellow powder.

NMR (D ₂ O) delta: 0.95 (3H, ie, J = 7.5 Hz), 1.25 (3H, d, 7.0 Hz), 2.05 (2H, sextet,

J = 7.5 Hz), 3.10 (2H, dd, J = 10 Hz, 2.5 Hz), 3.35 (1H, dd, J = 6.5 H 2.5 Hz), 4.0 to 4.8 (6H, m), 7.1 (2H, dj J = 6.0 Hz), and 7.80 (2H, d,

J = 6.0 Hz).

Ií (KBr technique) gamma _ma; ,: 3,400, 1,750 and 1,590 cm ¹ .

Analysis for C ₃ gH ₂₂ N ₉ O^S . 2H.,0 calculated 54.52 i C, 6.10% H, 7.07% N, found 54.32 * C, 6.03% H, 6.99% N.

Example 7

Preparation of 3-(N-methyl-3-methylpyridine-2-methanethio)-6alpha-Q-(R)hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

3-(N-methyl-3-methylpyridin-2-ylmethanethio)-6alpha-Q-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

To a solution of 680 mg (1.45 mmol) of compound 38 in 120 mL of methylene chloride was added 270 mg (2.33 mmol) of methyl fluorosulfonate. The reaction mixture was stirred for 3 h at room temperature. The precipitate was filtered off and washed with methylene chloride (5 mL) to give 840 mg (99% yield) of quaternized pyridine 39 as white crystals.

NMR (DMSO-d6) delta: 1.15 (3H, d, J = 5.8 Hz), 2.62 (3H, s), 3.2 to 4.4 (5H, m), 4.45 (3H, s), 4.60 and 4.82 (1H each, ABq, J - 9.2 1-Iz), 5.30 and 5.46 (1H each, ABq, J = 12.8 Hz) and 7.6 to 8.9 (7H, m).

IR (KBr technique) gamma^^^: 3,400, 1,750 and 1,590 cm \

Analysis for ^C 24 ^H 24 ^N 3°9 ^S 2 ^F calculated 49.14% C, 4.47% H, 7.13% N, 11.43% S, found 49.56% C, 4.16% H, 7.26% N, 11.03% S.

To a solution of 810 mg (1.39 mmol) of compound 39 in 100 ml of tetrahydrofuran and 100 ml of ether was added 100 ml of buffer pH 7.0 and then 750 mg of 10% palladium on carbon was added. The mixture was hydrogenated for 60 min at 310 kPa on a Paar shaker in a cold room (4 to 6 °C).

The reaction mixture was filtered through celite and the catalyst washed with ether (2 x 10 ml). The combined filtrate and washings were extracted with ether (2 x 40 ml) and lyophilized to give a yellow solid which was purified on a C^g BONDAPAK (Waters Associates) column (20 g), eluting with 5% acetonitrile in water at 55 kPa.

Each 15 ml fraction was assayed by high-performance liquid chromatography and fractions having a 11-fold absorption at lambda _max 300 nm were pooled and lyophilized to give 141 mg (30% yield) of the title compound as a yellow solid.

NMR (D ₂ O) delta: 1.24 (3H, d, J = 7.0 Hz), 2.62 (3H, s), 3.2 to 3.5 (3H, m), 4.2 to

4.4 (2H, m), 4.45 (3H, s), 4.50 and 4.59 (1H each, ABq, J = 12.16 Hz), 7.82 (1H, q, J ----- 7.0 Hz, 6.5 Hz), 8.35 (1H, d, J = 7.0 Hz) and 8.65 (1H, d, J = 6.5 Hz).

IR (KBr technique) gamma _max : 3,400, 1,750 and 1,580 cm h UV lambda _max (H ₂ O): 296 nm (epsilon = 8,014).

Example 8

Preparation of 3-(2-methyl-N-methylthiazol-4-ylmethanethio)-6-alpha-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-ene-2-carboxylate

OH

COjpNB

3-(2-Methyl-N-methylthiazol-4-ylmethanethio)-6alpha-fl-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

To a solution of 525 mg (1.1 mmol) of compound 9 in 20 mL of methylene chloride was added 0.27 mL (3.3 mmol) of methyl fluorosulfonate. The reaction mixture was stirred for 50 minutes at room temperature.

The precipitate was filtered off and washed with methylene chloride (50 ml) to give 650 mg (100% yield) of quaternized thiazole 10, which was used for the next step without further purification.

Then, to a solution of compound 10 in 100 ml of tetrahydrofuran and 100 ml of ether, 100 ml of pH 7.0 buffer was added, followed by the addition of 500 mg of 10% palladium on carbon. The mixture was hydrogenated at 241 241 kPa for 45 minutes on a Paar shaker.

The reaction mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL).

The combined filtrate and washings were extracted with ether (2 x 100 ml) and lyophilized to give a yellow powder which was purified on a BONDAPAK reversed phase column (8 g) (Waters Associates), eluting with 5% acetonitrile in water at 55 kPa.

Each 15 ml fraction was assayed by high-performance liquid chromatography and the fractions with an ultraviolet absorption lambda _{max of} 300 nm were pooled and lyophilized to give 145 mg (48% yield) of the title compound as a pale yellow powder.

NMR (CDCl ) delta: 1.20 (3H, d, J = 7 Hz), 2.92 (3H, s), 3.08 (1H, d, J = 3.5 Hz), 3.20 (1H, d, J = 3 Hz), 3.44 (1H, dd, J = 1 Hz, J = 3.5 Hz), 4.00 (3H, 5), 4.20 (3H, m), 4.36 (2H, m) and 7.88 (1H, s).

IR (KBr technique) gamma: 3,400, 1,750 and 1,585 cm

UV lambda _max (H^O) ^: nm (epsilon = 7,500) .

Analysis for ^c i5 ^H i8 ^N 2°4 ^S 2* ^2H 2° calculated 46.15% C, 5.64% H, 7.17% N, 16.41% S, found 46.50% C, 5.26% H, 7.13% N, 16.20% S.

Example 9

Preparation of 3-(N,N'-dimethylimidazol-2-ylmethanethio)-6alpha-Q-(R)-hydroxyethyl^-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

3~ (Ν,N'-dimethylimidazol-2-ylmethanethio)-6alpha-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

To a suspension of 1.34 g (3.0 mmol) of compound 33 in 270 ml of acetone was added 20 ml of methyl iodide. The reaction mixture was stirred for 4 days at room temperature. The precipitate was filtered off and washed with acetone (20 ml) to give 1.70 g (96% yield) of quaternized imidazole 34 as yellow crystals, mp 175-177 °C.

NMR (DMSO-d6) delta: 1.10 (3H, d, H = 6.2 Hz), 3.30 (2H, s), 3.2 to 4.3 (6H, m), 3.95 (6H, s), 5.45 (2H, ABq, J = 14 Hz), 7.65 (2H, d, J = 6.0 Hz).

IR (KBr technique): ny _mav : 3,400, 1,750 and 1,600 cm ^-1 .

* IÍIQA

Analysis for ^C 21 ^H 22 ^N 4°6 ^S 1 calculated 43.08% C, 9.60$ H, 5.48% N, found 43.02% C, 9.02% H, 5.44% N.

To a solution of 1.30 g (1.86 mmol) of compound 34 in 120 ml of tetrahydrofuran and 120 ml of ether was added 120 ml of pH 7.0 buffer, followed by the addition of 900 mg of 30% palladium on celite. The mixture was hydrogenated for 40 minutes at 276 kPa on a Paar shaker.

The mixture was filtered through celite and the catalyst was washed with water (2 x 15 ml). The combined filtrate and washings were extracted with ether (3 x 100 ml) and lyophilized to give a yellow amorphous powder which was purified on a C^g BONDAPAK (Waters Associates) column (30 g) eluting with 10% acetonitrile in water at 55 kPa.

Each 200 ml fraction was assayed by high-performance liquid chromatography and the fractions with ultraviolet absorption at lambda _max 300 nm were pooled and lyophilized to give 220 mg (35% yield) of the title compound as a yellow powder.

NMR ( _D2O ) delta: 1.12 (3H, d, J = 7.0 Hz) , 3.08 (1H, dd, J = 13, ,0 Hz, 6.4 Hz). , 3.15 (1H, dd, J = 13.0 Hz, . 6.4 Hz), 3.45 (1H , dd, , J = 3.2Hz, 4.5 Hz) , 3.85 (6H, s), 4.1 and 4.3 (2H, m), 4, 40 (1H, d, J = 13.5 Hz), 4.52 (1H, d, J = 13.5 Hz), , and 7 .40 (2H, sec)

IR (KBr technique) gamma^^^: 3,500, 1,750 and 1,590 cm L UV lambda _max (HjO) ^: 296 nm (epsilon = 8,411).

Analysis for C gN^O^S. í^O calculated 51.68% C, 5.67% H, 12.06% N, 9.50 i S, found 49.93% C, 5.94% H, 11.46% N, 9.03% S.

Example 10

Preparation of 3-(2,3,4-trimethylthiazol-5-ylmethanethio)-6alpha-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

OH

Pd/C, H ₂

Ψ

3-(2,3,4-trimethylthiazol-5-yl-methanethio)-6alpha-El-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

To a solution of 1.97 g (4.0 mmol) of compound 48 in 180 ml of methylene chloride was added a solution of 0.98 ml (13 mmol) of methyl fluorosulfonate in 2 ml of methylene chloride. The reaction mixture was poured into a solution of ether (400 ml) and n-pentane (100 ml). The precipitate was filtered off and washed with ether (20 ml), to give 1.6 g (65.5% yield) of quaternized thiaazole 49 as a white amorphous powder.

NMR (DMSO-d6) delta: 1.25 (3H, s, J = 6.5 Hz), 2.45 (3H, s), 2.80 (3H, s), 3.2 to 4.5 (6H, m), 3.90 (3H, s), 5.30 (2H, broad), 7.60 and 8.2 (1H, d, J - 8.5 Hz each).

IR (KBr technique) 3,400, 1,770 and 1,690 cm \

Analysis for ^23 ^H 26 ^N 3°9 ^S 3 ^F '' ^H 2° calculated 45.09% C, 4.44% H, 6.86% N, found 44.50% C, 4.38% H, 6.58% N.

To a solution of 1.0 g (1.72 mmol) of compound 49 in 100 mL of tetrahydrofuran and 100 mL of ether was added 100 mL of buffer 7.0, followed by the addition of 1.0 g of 10% palladium on carbon. The reaction mixture was hydrogenated for 40 minutes at 276 kPa on a Paar shaker.

The mixture was filtered through celite and the catalyst washed with water (2 x 10 ml). The combined filtrate and washings were extracted with ether (3 x 100 ml) and lyophilized to give a yellow powder which was purified on a C-^θ BONDAPAK (Waters Associates) column (40 g), eluting with 10% acetonitrile in water at 55 kPa.

Each 15 ml fraction was assayed by high-performance liquid chromatography and the fractions with ultraviolet absorption at lambda _max 30 nm were pooled and lyophilized to give 315 mg (50% yield) of the title compound as a yellow solid.

NMR (D ₂ O) delta: 1.25 (3H, d, J = 7.0 Hz), 2.25 (3H, s), 2.90 (3H, s), 3.0 to 3.30 (3H, m), 3.90 (3H, s) and 4.1 to 4.4 (4H, m).

ID (KBr technician^ 9 ^ama max ^: 3,400, 1,750 and 1,580 cm \

UV lambda _max 1^0: 297 nm (epsilon = 8,994).

Analysis for C-12.2O calculated 48.25% C, 6.09% H, 7.79% N, found 47.96% C, 5.83% H, 7.89% N.

Example 11

Preparation of 3-[2-(N-methylthiazoliummethylthio]-6-alpha(1-(R)-hydroxyethyl)-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

3-[2-(N-methylthiazoliummethylthio]-6alpha-(j.(R)-hydroxyethyl)-7-oxo-1-azabicyclo(3,2,0)hept-2-ene~2-carboxylate

To a solution of 782 g (1.36 mmol) of compound 6 in 55 mL of methylene chloride was added 0.5 mL of methyl sulfonate and stirred at room temperature for 90 min. The precipitate was filtered off, washed with methylene chloride (30 mL) and ether (20 mL) to give 630 mg of crude quaternized thiazole 7, which was used for the next step without further purification.

To a solution of compound 7 in 140 ml of tetrahydrofuran and 120 ml of ether was added 140 ml of pH 7.0 buffer, followed by the addition of 650 mg of 10% palladium on carbon. The reaction mixture was hydrogenated for 35 minutes at 207 kPa on a Paar shaker.

The mixture was filtered and the catalyst washed with water (2 x 10 ml). The combined filtrate and washings were extracted with ether (2 x 150 ml) and lyophilized to give a yellow powder. The crude yellow powder was purified on a BONDAPAK reversed phase column (7 g) (Waters Associates), eluting with 5% acetonitrile in water at 55 kPa.

Each 15 ml fraction was tested by high-performance liquid chromatography, the fractions with ultraviolet absorption at lambda _max 300 nm were pooled and lyophilized to give 23 mg (5% yield) of the title compound as a yellow amorphous solid.

NMR (D ₂ O) delta: 1.28 (3H, d, J = 7.0 Hz), 3.12 (2H, d, J = 7.0 Hz), 3.44 (1H, dd,

J = 1.0 Hz and 3.0 Hz), 4.20 (3H, s), 4.24 (2H, m), 4.76 (3H, m), 8.12 (1H, d, J ~ 4 Hz) and 8.24 (1H, d, J = Hz).

IR (KBr technique) gamma: 3,400, 1,740 and 1,580 cm \ ³ max.

UV lambda (H _n O): 292 nm (epsilon = 7,285). max 2 ^years

Example 12

Preparation of 3-fl-(RS)-methyl-N-methylpyridin-3-yl-methanethiol-6alpha-JjL — (R >-hydroxyethyl] -7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

3- [~1~ (RS)-methyl-N-methylpyridin-3-yl-methanethioj-6alpha-Q-(R)-hydroxyethylj-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

To a solution of 1.1 g (2.34 mmol) of compound 28 in 100 mL of acetone was added 10 mL of methyl iodide. The reaction mixture was stirred at room temperature for 18 h. The precipitate was filtered off and washed with methylene chloride (10 mL) to give 1.4 g (100% yield) of quaternized pyridine 29 as a yellow powder.

NMR (DMSO“d 6 ) delta: 1.10 (3H, d, J = 6.5 Hz), 1.62 (3H, d, J = 7.5 Hz'), 2.6 to 4.2 (6H, m), 4.39 (3H, s), 5.42 (2H, ABq, J = 13.6 Hz) and 7.9 to 9.2 (8H, m),

IR (KBr-technique) ⁿ tnax ^: 2,400, 1,770 and 1,190 cm

Analysis for ^C 24 ^H 26 ^N 3°6 ^S 1 ^I 1 calculated 47.14% C, 4.29 ΐ H, 6.87% N, 5.24 $ S, found 47.15 í C, 4.78% H, 6.11% N, 5.41 t S.

To a solution of 1.45 g (2.37 mmol) of compound 29 in 120 ml of tetrahydrofuran and 120 ml of ether was added 120 ml of buffer pH 7.0, followed by the addition of 1.5 g of 10% palladium on carbon. The reaction mixture was hydrogenated for 60 minutes at 310 kPa on a Paar shaker. The mixture was filtered through celite and the catalyst was washed with water (2 x 15 ml).

The combined filtrate and washings were extracted with ether (2 x 200 ml) and lyophilized to give a yellow solid which was purified on a C18 BONDAPAK (Waters Associates) reversed phase column (50 g), eluting with 5% acetonitrile in water at 55 kPa.

Each 20 ml fraction was assayed by high-performance liquid chromatography and the fractions with ultraviolet absorption at lambda _max 300 nm were pooled and lyophilized to give 200 mg (24% yield) of the title compound as a yellow amorphous solid.

NMR (D ₂ O) delta: 1.32 (3H, d, J = 7.0 Hz), 1.63 (3H, d, J = 7.2 Hz), 2.5 to 4.6 (6H, m), 4.32 (3H, s), and 8.2 to 8.9 (4H, m).

IR (KBr technique) ~ 'new _3χ : 3,400, 1,750 and 1,590 cm ^-1 .

UV lambda (H„0): 296 nm' (epsilon = 7573). max 2

Analysis for ^37 ^Η 20 ^Ν 2°4 ⁸ 1^^^ ^2Η 2θ calculated 54.38% C, 5.77% H, 7.46% N, found 54.39% C, 5.98% H, 7.68% N.

Example 13

Preparation of 3-(N-methyl-N'-benzylimidazol-2-ylmethanethio)-6alpha-[j-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

₄₃ COjpNB

4!

3-(N-methyl-[T]-benzylimidazol-3-ylmethanethio)-6alpha-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

To a solution of 1.76 g (3.3 mol) of compound 43 in 1.1 L of methylene chloride was added 1.15 ml (13.4 mmol) of methyl fluorosulfonate. The reaction mixture was stirred for 2 h at room temperature.

The reaction mixture is concentrated in vacuo to a volume of about 15 ml.

The precipitate was filtered off, washed with methylene chloride (10 ml) to give 1.58 g (74% yield) of quaternized imidazole 44 as a white solid.

NMR (DMSO-d6) delta: 1.15 (3H, d, J = 7.0 Hz), 3.2 to 4.4 (6H, m), 4.70 and 5.0 (1H each, ABq, J = 10.8 Hz), 5.24 and 5.46 (1H each, ABq, J = 14 Hz), 5.50 (2H, s) and 7.4 to 8.4 (11H, m).

ID (KBr technique) ^{maximum :} 500, _1,770 and 1,700 cm ¹ .

Analysis for ^C 28 ^H 29 ^N 4°9 ^S 2 ^F calculated 51.48% C, 4.47% H, 8.67% N, 10.20% S, found 51.84% C, 4.52% H, 8.65% N, 9.87% S.

To a solution of 1.11 g (1.71 mmol) of compound 44 in 100 ml of tetrahydrofuran and 100 ml of ether was added 120 ml of pH 7.0 buffer, followed by the addition of 1.0 g of 10% palladium on carbon. The mixture was hydrogenated for 45 minutes at 310 kPa on a Paar shaker.

The mixture was filtered through celite and the catalyst washed with water (2 x 10 ml). The combined filtrate and washings were extracted with ether (2 x 70 ml) and lyophilized to give a yellow powder which was purified on a BONDAPAK (Waters Associates) column (40 g), eluting with 10% acetonitrile in water at 55 kPa.

Each 15 ml fraction was assayed by high-performance liquid chromatography, and the fractions with ultraviolet absorption at lambda _χ 300 nm were pooled and lyophilized to give 305 mg (43% yield) of the title compound as a pale yellow amorphous solid.

NMR (DMSO) delta: 1.40 (3H, d, J = 7.0 Hz), 2.9 to 3.4 (3H, m), 3.98 (3H, s), 4.0 to

4.2 (2H, m), 4.23 (2H, broad s), 5.57 (2H, s), and 7.2 to 7.65 (7H, m).

IR (KBr-technique) ⁿ ýmax ^: 400, 1,760 and 1,590 cm \

UV lambda (H^O) : 299 nm (epsilon = 8807).

Analysis for ^C 21 ^H 23 ^N 3°4^1 ' ^{4 4H} 2° calculated 57.25% C, 5.94% H, 9.54% N, 7.28% S, found 56.66% C, 5.70% H, 9.49% N, 8.30% S.

Example 14

Preparation of 3-(2-methyl-N-methylpyridin-3-ylmethanethio)-6alpha-[^1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate

3-(2-Methyl-N-methylpyridin-3-ylmethanethio)-6alpha-[1-(R)-hydroxyethyl]-7-oxo-1-azabloycyclo(3.2.0)hept-2-ene-2-carboxylate

To a solution of 697 mg (1.19 mmol) of compound 16 in 100 mL of methylene chloride at 10 °C was added dropwise 0.5 mL (6.18 mmol) of methyl fluorosulfonate over 10 min. The mixture was stirred at room temperature for 2.5 h. The precipitate was filtered off and washed with 30 mL of methylene chloride to give 777 mg (90% yield) of quaternized pyridine 17 as a yellow solid.

NMR (CDCl1) of compound 17 delta: 1.20 (3H, d, J = 7 Hz), 2.82 (3H, s,), 4.36 (3H, s), 4.16 (2H, m), 4.60 (2H, s), 5.20 (1H, m), 5.42 (2H, q, J = 14 Hz), 7.80 (2H, d, J = 8 Hz), 8.04 (1H, dd,

J = 7 Hz, 6.5 Hz), 8.32 (2H, d, J = 8 Hz), 8.64 (1H, d, J = 7.5 Hz), and 9.08 (1H, d, J % 7.5 Hz).

ID number (KBr technique) ⁿ y _ma x ^! 3,500 and 1,765 cm ⁹ .

Analysis for ^C 24 ^H 26 ^FN 3°9 ^S calculated 48.91% C, 4.55% H, 7.23% N, 11.04% S, found 49.39% C, 3.97% H, 7.20% N, 10.98% S.

To a solution of 1.10 g (1.88 mmol) of compound 17 in 80 ml of tetrahydrofuran and 80 ml of ether was added 80 ml of pH 7.0 buffer, followed by the addition of 800 mg of 10% palladium on carbon. The mixture was hydrogenated for 40 minutes at 207 kPa on a Paar shaker.

The mixture was filtered through celite and the catalyst washed with water (2 x 10 ml). The combined filtrate and washings were extracted with ether (2 x 100 ml) and lyophilized to give a yellow powder which was purified by chromatography on a HP.20 column, eluting with water and 5% acetonitrile in water. Each 15 ml fraction was tested by high pressure liquid chromatography and the fractions with ultraviolet absorption at lambda _max 300 nm were combined and lyophilized to give 614 mg (42% yield) of the title product as a pale yellow powder.

NMR (D ₂ O) delta: 1.28 (d, 3H, J = 7 Hz), 2.86 (3H, s), 3.20 (2H, dd, J = 10 Hz, 3.5 Hz), 3.42 (1H, dd, J = 5.4 Hz, 3.5 Hz), 4.20 (3H, m), 4.32 (3H, s), 4.35 (2H, s), 9.88 (1H, dd, J = 7.2 Hz, 6.5 Hz), 8.5 (1H, d, J = 8 Hz), and 8.7 (1H, d, J = 8 Hz).

IR (KBr technique) gamma _max : 3,400, 1,760 and 1,590 cm L UV lambda _max (H ₂ 0): 298 nm (epsilon = 8,391).

Analysis for C ₁₇ H ₂₀ N ₂ O ₃ SH ₂ o calculated 55.73% C, 5.46% H, 7.65% N, 8.74% S, found 55.50% C, 6.05% H, 7.74% N, 8.68% S.

Example 15

Alternative procedure for the preparation of ethyl] -7-oxo-1-aza-bicyclo[3-(N-methylpyridin-2-ylmethanethio)-6alpha-[1-(R)-hydroxy 2.0)-hept-2-ene-2-carboxylate

To a solution of allyl ester (1.79 g, 4.97 mmol), tetrakistriphenylphosphinepalladium (175 mg, 0.15 mmol), and triphenylphosphine (175 mg, 0.67 mmol) in methylene chloride (25 mL) was added a solution of potassium 2-ethylhexanoate (1.085 g, 5.96 mmol) in ethyl acetate (12 mL).

The mixture was stirred at room temperature for 1 hour, after which only traces of starting material were detected by thin layer chromatography. The reaction mixture was diluted with anhydrous diethyl ether (150 mL) and the precipitate was filtered off, washed with ethyl acetate and then ether to give a light brown solid.

This material was dissolved in water (10 mL) and purified by reverse phase chromatography to give 1.85 g of the title compound as a cream colored solid. This was further purified by trituration in acetone to give 1.47 g (83% yield) of the pure title compound.

¹ H-NMR (D ₂ O) delta: 8.45 to 8.36 (m, 1H), 7.92 to 7.22 (m, 3H), 4.78 to 3.91 (m, 2H), 4.69 (s, 2H), 3.34 to 2.71 (m, 3H), 1.19 (d, J = 6.4, 3H).

To a cooled suspension (0 °C) of potassium 6-hydroxyethyl-2-(2-pyridylmethylthio)carbapenem-3-carboxylate (53.8 mg, 0.15 mmol) in acetone (2 mL) was added toluenesulfonic acid (27.6 (27.6 mg, 0.16 mmol).

The reaction mixture was stirred for 20 min at 0°C and then methyl triflate (0.02 mL) was added. The mixture was stirred for 60 min at 0°C and then LA-1 resin was added followed by hexane (6 mL). The reaction mixture was extracted with water (4 x 0.5 mL) and the combined aqueous phases were purified by high pressure liquid chromatography and reverse phase to give 10 mg of the title compound.

Example 16

Preparation of (5R,6S)-3-(dimethylpyridinium-4-yl)methyl)thio]-6-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate (16A) and

(4R,5R,6S)-3-[[(1,33dimethylpyridinium-4-yl)methyl]thio]-6-[1-(R)-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo[3,2,0]hept-2-ene-2-carboxylate (16B)

16A R = H 16B R = CH ₃

A. Preparation of 4-hydroxymethyl-3-methylpyridine

procedure according to Boekelheid (V. Boekel

1) h ₂ o ₂

--2) Ac ₂ 0

3) H ₃ O ®

For the preparation of hydroxymethylpyridine, see Heide, W. J. Linn, JACS, 76, 1286 (1954). A freshly distilled solution of 3,4-lutidine (46.0 g, 0.43 mol) in 120 ml of glacial acetic acid was cooled to 0°C and then 64 ml of 30% hydrogen peroxide was added dropwise.

The resulting solution is heated for 3 hours at 75 °C in an oil bath. Then another 20 ml of 30% hydrogen peroxide is added and heating is continued for 18 hours. Finally, another 20 ml of 30% hydrogen peroxide is added and the reaction mixture is maintained at 75 °C for another 3 hours. The solution is then concentrated to about 100 ml, 50 ml of water is added and the mixture is concentrated to about half its volume. The resulting mixture is cooled (0-5 °C) and basified to about pH 10 using cold 40% aqueous sodium hydroxide solution.

The mixture was then extracted with methylene chloride (5x) and the extract was dried (Na ₂ CO ₃ + Na ₂ SO ₄ ) and concentrated on a rotary evaporator to give a yellow solution. Dilution of this solution with hexane gave a solid which was filtered off and dried in vacuo.

3,4-lutidine-N-oxide (48.0 g, 83%) was obtained as an off-white solid.

The N-oxide was added portionwise to 60 ml of acetic anhydride and the resulting dark orange solution was heated on a water bath at about 90 °C for one hour. The excess acetic anhydride was distilled off under reduced pressure to give a substance boiling at 90-120 °C/13.3 Pa (39.0 g). Chromatography of this oil (silica gel/ethyl acetate-petroleum ether 2:3) gave pure 4-acetoxymethyl-3-methylpyridine (19.0 g, 30%) as an oil: ir (oil) 1745 cm- ¹ .

Acetate was added to 100 ml of 10% aqueous hydrochloric acid and heated to boiling for 1 hour. The resulting solution was cooled to 0°C, basified with solid potassium carbonate, and then extracted with methylene chloride (3 x 100 ml).

The organic extract was washed with brine, dried over sodium sulfate, and evaporated to give 11.0 g of an off-white solid, mp 70-72°C. The solid was triturated with cold ether to give pure 4-hydroxymethyl-3-methylpyridine (9.5 g, 67%) as a white solid, mp 77-80°C (according to W. L. F. Armarego, B. A. Milloy, S. C. Sharma, JCS, 2485 (1972) mp 81-82°C).

HNMR (CDCl ₃ ) delta: 8.27, 7.41 (ABq, J = 5 Hz, 2H), 8-.18 (s, 1H), 5.63 (broad s, -OH), 4.67 (s, CH ₂ ), 2.20 (s, CH ₃ ); iC (nujol) 3 170 cm” ¹ .

B. Preparation of 4-(acetylthiomethyl)-3-methylpyridine

To an ice-cooled solution of triphenylphosphine (17.04 g, stirred with a mechanical stirrer)

0.065 mol) in 250 ml of anhydrous tetrahydrofuran was added dropwise diisopropylazodicarboxylate (12.8 ml, 0.065 mol) and the resulting suspension was stirred at 0 °C for 1 hour.

To this mixture was added dropwise a solution of 4-hydroxymethyl-3-methylpyridine (4.0 g, 0.0325 mol) in 100 ml of anhydrous tetrahydrofuran, followed by freshly distilled thioacetic acid (4.64 ml, 0.065 mol).

The resulting mixture was stirred at 0°C for 1 hour and then at room temperature for 1 hour to give an orange solution. The solution was concentrated on a rotary evaporator and then diluted with petroleum ether. The resulting mixture was filtered and the filtrate was evaporated to give an orange oil.

Chromatography (silica gel/hexane then 10%-50% ethyl acetate/hexane) of this oil gave 7.0 g of oil which was distilled (knuckle condenser) to give the pure product (6.0 g, 100%) as a yellow oil, b.p. (air bath temperature) 95-100°C/13.3 Pa.

¹ HNMR (CDCip delta: 8.40, 7.20 (ABq, J = 5 Hz, 2H) , 8.37 (s, 1H) , 4.08 (s, CHp , 2.35 (s, CH ₃ ), 2.32 (s, CH ₃ ); iC (oil) 1695 cm ^-1 ).

C. Preparation of 4-acetylthiomethyl)-1,3-dimethylpyridinium trifluoromethanesulfonate

To an ice-cooled solution of ghiolacetate (2.95 g, 0.016 mol) in 10 ml of methylene chloride was added dropwise methyl trifluoromethanesulfonate (4.60 ml, 0.04 mol) and the mixture was stirred at 0 °C under nitrogen for 1 hour. The reaction mixture was then evaporated to dryness and the residue triturated in ether.

The resulting solid was filtered off and dried in vacuo to give the product (4.0 g, 72%) as a white solid.

¹ HNMR (CDCl ₃ ) delta: 8.72 (s, 1H) , 8.58 , 7.87 (ABq, J = 6 Hz, 2H) , 4.39 (s, N-CH-j) ,

4.17 (s, CH ₂ ), 2.53 (s, CH 2 ), 2.36 (s, CH 2 ); IR (oil) 1,700 cm ^-1

D. Preparation of (5R,6S)-3-([(1,3-dimethylpyridinium-4-yl)methyl]thio)-6-[beta]-(R)-hydroxyethyl]-7-oxo-1-azabicyclo[3,2,θ]hept-2-ene-2-carboxylate

_CH2SAc

CH,

1) NaOH/ _H2O

OH

I ch ₃

2) io ] ýy—ορ(οφ) ₂ CO ₂ pNB

OH

3) _H2 , Pd/C

CH,

WHAT,

Nx/

CH,

To an ice-cold, nitrogen-flushed solution of sodium hydroxide (0.324 g, 0.008 mol) in 10 ml of water was added thioacetate (1.40 g, 0.004 mol) and the mixture was stirred at 0 °C under nitrogen for 1 h. The pH was then adjusted to 7.2-7.3 with 10% aqueous potassium dihydrogen phosphate solution and the resulting solution was added dropwise to an ice-cold solution of enol phosphate (1.45 g,

0.0025 mol) in 20 ml of tetrahydrofuran. The mixture is stirred at 0 °C for 1 hour and then transferred to a pressure vessel.

To this mixture were added 20 ml of ether, 25 ml of 0.1M phosphate buffer (pH 7.4) and 1.4 g of 10% palladium on carbon. The mixture was then hydrogenated at 310 kPa for 1 hour. The reaction mixture was filtered through Celite and washed with ether and phosphate buffer pH 7.4.

The aqueous phase was separated and the remaining solvents were removed in vacuo. The resulting aqueous solution was loaded onto a reversed phase column (C18 Bondapak) which was eluted with water and then 10% acetonitrile in water.

Lyophilization of the appropriate fractions yielded 0.9 g of an orange solid. This was rechromatographed using water and then 2% acetonitrile in water as eluent.

Lyophilization afforded pure compound 16A (0.25 g, 57%) as a yellow solid. ¹ HNMR (θ ₂ Ο) delta 8.55 (s, 1H), 8.53, 7.96 (ABq, J = 6.8 Hz, 2H), 4.30-3.99 (m, 2H), 4.27 (s, 5H), 3.35 (dd, = 2.8 Hz, J ₂ - 6.0 Hz, 1H), 3.05 (d, J = 8.8 Hz, 2H), 2.58 (s, 3H), 1.23 (d, J = 6.3 Hz, 3H); IR (KBr) 1,755, 1,590 cm ; UV (phosphate buffer, pH 7) 295 nm (ε 7,180).

E. Preparation of (4R,5R,6S)-3-[1-(3-dimethylpyridinium-4-yl)-methyl]thiol-6-[1-(R)-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo 3,2,0 hept-2-ene-2-carboxylate

3) _H2 , Pd/C

To an ice-cooled solution of bicyclic ketone (0.906 g, 0.0025 mol) in 10 ml of acetonitrile was added diphenylchlorophosphate (0.544 ml, 0.00263 mol), diisopropylethylamine (0.457 ml, 0.00263 mol) and 4-dimethylaminopyridine (0.3 mg) sequentially.

After 50 minutes, the reaction mixture was diluted with cold ethyl acetate and then washed with cold water and brine. The organic phase was dried (Na2SO4) and evaporated at room temperature to give the enol phosphate as an off-white foam. This foam was taken up in 20 mL of tetrahydrofuran, cooled to -30°C under nitrogen, and then treated with an aqueous solution of the thiolate [previously prepared from 0.3 g of sodium hydroxide (7.5 mmol) and 1.3 g of thioacetate (3.76 mmol) in 10 mL of water].

The reaction mixture was stirred at -30°C for 30 minutes, then at 0°C for 75 minutes and finally transferred to a pressure vessel containing 20 ml of ether, 30 ml of 0.1M phosphate buffer (pH 7.4) and 1.5 g of 10% palladium on carbon.

After hydrogenation at 310 kPa for 1 hour, the mixture is filtered through Celite and the aqueous phase is separated and concentrated in vacuo. The resulting solution is applied to a reversed phase column (C ₁₀ Bonda1 o pak) which is eluted with water.

Lyophilization of the appropriate fractions yielded 1.2 g of a yellow solid. This was rechromatographed (elution with water up to 4% acetonitrile and water) to yield, after lyophilization, pure compound 16B (0.250 g, 28%) as a yellow solid.

¹ HNMR (D ₂ O) delta 8.53 (s, 1H) , 8.49, 7.81 (ABq, J = 6.2 Hz, 2H) , 4.38 to 3.98 (m, 4H) , 4.27 (s, 3H), 3.49 to 3.18 (ra, 2H), 2.51 (s, 3H), 1.25 (d, J = 6.7 Hz, 3H), 1.16 (d,

J = 7.6 Hz, 3H)1 IR (KBr) 1750, 1595 cm -1; UV (phosphate buffer, pH 7) 292 nm (epsilon 7930).

Example 17

Preparation of (5R, 6S)-3- f [(1,2-dimethylpyridinium-4-yl)methyl]thio]-6- Ll-(R)-hydroxyethyl]-7-oxo-1-azabicyclo[3,2,0]hept-2-ene-2-carboxylate (17A) (4R,5R,6S)-3- Ll(1,2-dimethylpyridinium-4-yl)methyl]thio]-6-[l-(R)-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo[3,2,0]hept-2-ene-2-carboxylate (17B)

To a suspension of 95% lithium aluminum hydride (2.4 g, 0.06 mol) in 150 ml of anhydrous ether is added a solution of methyl 2-methylisonicotinate (14.0 g, 0.093 mol, prepared according to 0. Efimovsky, P. Rumpf, Bull. soc. chim. Fi., 648 (1954)) in 50 ml of anhydrous ether at -5 °C in a nitrogen atmosphere.

The resulting mixture was stirred for 30 minutes at room temperature and then refluxed for 2 hours. A further 1.2 g (0.03 mol) of lithium aluminium hydride was added portionwise and refluxed for a further 1 hour.

A further 1.2 g (0.03 mol) of lithium aluminium hydride is added portionwise and refluxed for a further 1 hour. The reaction mixture is then cooled to 0°C and 3.75 ml of water, 3.75 ml of 15% aqueous sodium hydroxide solution and finally 11.25 ml of water are added successively.

The suspension is then filtered and the filter cake is washed with ether and then with ethyl acetate. The filtrate is evaporated to give a dark yellow oil which is taken up in acetonitrile and then filtered through a silica gel column (elution with acetonitrile and then acetone).

The product (7.7 g, 67%) was obtained as a yellow oil. ³ HNMR (CDCl^) delta 8.30, 7.10 (ABq, J = 5 Hz, 2H), 7.17 (s, 1H), 5.42 (s, -OH), 4.70 (s, CHp , 2.50 (s, CH-j) . ¹

B. Preparation of 4-(acetylthiomethyl)-2-methylpyridine

ch ₂ sac ch ₃

To a solution of triphenylphosphine (31.4 g, 0.12 mol) in 200 ml of anhydrous tetrahydrofuran under nitrogen at -5 °C was added dropwise diisopropylazodicarboxylate (23.6 ml, 0.12 mol) and the mixture was stirred at -5 °C for 1 hour.

To the resulting suspension was added a solution of 4-hydroxymethyl-2-methylpyridine (7.60 g, 0.062 mol) and freshly distilled thioacetic acid (8.60 ml, 0.12 mol) in 40 ml of anhydrous tetrahydrofuran over about 10 minutes.

The reaction mixture was stirred for 30 minutes at 0°C and then for 1 hour at room temperature. The resulting suspension was filtered and the filtrate was concentrated to give an orange-yellow liquid which was diluted with ether and filtered.

The filtrate was evaporated and the residual oil was chromatographed (silica gel/ethyl acetate-hexane 1:1) to give the thioacetate (8.87 g, 79%) as a yellow oil. ⁴ HNMR (CDCl^) delta 8.45,

7.03 (ABq, J = 5 Hz, 2H), 7.08 (s, 1H), 4.04 (s, CH ₂ ), 2.55 (s, CHj ), 2.39 (s, CHj ) IC (oil) 1695 cm ¹ .

C. Preparation of 4-mercaptomethyl-2-methylpyridine ch ₂ sh ох ^N ch ₃

To 15 ml of ice-cold, nitrogen-flushed 1N sodium hydroxide solution was added 4-acetylthiomethyl-2-methylpyridine (1.358 g, 0.0075 mol) in one portion. After stirring at 0 °C for 15 min, the reaction mixture was washed with ether (2 x 5 ml), neutralized with concentrated hydrochloric acid, and extracted with methylene chloride (3 x 10 ml).

Evaporation of the methylene chloride solution gave the thiol (0.89 g, 96%) as a yellow oil, which gradually turned pink on standing. ^HNMR (CDCl^) delta 8.43, 7.37 (ABq, J = 5 Hz, 2H),

7.43 (s, 1H), 3.63 (d, J = 7.5 Hz, CH ₂ >, 2.55 (s, CH ₃ ), 1.81 (t, J = 7.5 Hz, SH).

D. Preparation of allyl-(5R,6S)-3-[_[(2-methylpyridin-4-yl)methyl]thio]-6-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo[3,2,oj hept-2-ene-2-carboxylate

OH

To an ice-cooled solution of the bicyclic ketone (0.760 g, 0.003 mol) in 8 ml of acetonitrile was added successively diphenylchlorophosphate (0.653 ml, 0.00315 mol), diisopropylethylamine (0.550 ml, 0.00315 mol) and dimethylaminopyridine (0.8 g).

The mixture was stirred at 0°C for 1 hour and then cooled to -20°C and 4-mercaptomethyl-2-methylpyridine (0.620 g, 0.00446 mol) was added followed by diisopropylethylamine (0.550 mL, 0.00315 mol). The reaction mixture was stirred at -20°C for 1.5 hours and then allowed to warm to room temperature.

The resulting mixture was diluted with 50 ml of ethyl acetate, washed with water, saturated sodium bicarbonate solution and saturated ammonium chloride solution, dried over sodium sulfate and evaporated.

The residue was chromatographed on silica gel (elution with ethyl acetate and then acetonitrile) to give a solid which was triturated in ether to give the pure product (0.820 g,

%) in the form of a white solid.

^X HNMR (CDCip delta 8.45, ^.09 (ABq, J = 5 Hz, 2H), 7.15 (s, 1H), 6.25 to 5.80 (m, 1H),

5.60 to 5.20 (m, 2H) , 4i.82 to 4.68 (m, 2H) , 4.55 to 4.05 (m, 2H) , 3.97 (s, 2H) , 3.16 to 2.93 (m, 3H), 2.55 (s, 3H), 1.84 (broad s, 1H), 1.32 (d, J = 6 Hz, 3H); ID (oil) 1 777,

695 cm ^-1 .

E. Preparation of (5R,6S)-3-[(1,2-dimethylpyridinium-4-yl)methyl]thiol-6-[1-(R)-hydroxyethyl]-7-oxo-1-azabiocyclo[3,2,o]hept-2-ene-2-carboxylate

A solution of the allyl ester (0.350 g, 0.936 mmol) in 6 mL of anhydrous acetonitrile was cooled to -5 °C and then methyl trifluoromethanesulfonate (0.111 mL, 0.983 mmol) was added. After 15 min, a solution of tetrakis(triphenylphosphine)palladium (0.027 g, 2.5 mol%) and triphenylphosphine (0.027 g) was added.

The reaction mixture was stirred for 5 minutes and then pyrrolidine (0.082 mL, 0.983 mmol) was added dropwise.

A solid slowly begins to separate from the resulting brown solution. The mixture is stirred vigorously for 20 minutes at 0 °C, then 15 ml of cold (0 °C) acetone is slowly added and stirring is continued for 20 minutes at 0 °C.

The resulting suspension was filtered and the residue was washed with cold acetone and dried in vacuo to give 0.345 g of a beige powder. This material was taken up in a small amount of phosphate buffer pH 7 (0.05M) and transferred to a short reversed phase column (C18 Bondapak).

Elution with water and lyophilization of the appropriate fractions gave 0.255 g of a pale yellow solid. This material was rechromatographed as before to give (after lyophilization) pure 17A (0.195 g, 60%) as a pale yellow solid.

^X HNMR (D ₂ O) delta 8.58, 7.83 (ABq, J = 6.4 Hz, 2H), 7.87 (s, 1H), 4.32 to 3.95 (m, 2H), 4.22 )s, 2H), 4.17 (s, 3H), 3.32 (dd, = 2.6 Hz, J ₂ = 6.1 Hz, 1H), 3.06 to 2.93 (m, 2H), 2.74 (s, 3H), 1.22 (d, J = 6.4 Hz, 3H); IR (KBr) 1,757, 1,590 cm ^-1 ; UV (phosphate buffer, pH 7.4) 296 nm (epsilon 7446).

F. Preparation of allyl-(4R,5R,6S)-3-[(2-methylpyridin-4-yl)methyl]thiol-6-p-(R)-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate

A solution of the alpha-diazoester (1/50 g, 0.00508 mol) in 12 ml of ethyl acetate and hexane (3:1) is heated to gentle reflux under nitrogen and then 0.020 g of rhodium octanoate is added all at once. Rapid evolution of nitrogen occurs for about 5 minutes and after boiling for a further 10 minutes the reaction is complete (thin layer chromatography).

The solvents were then removed under reduced pressure and the remaining gum was taken up in 15 mL of acetonitrile. The solution was cooled to -5°C and treated with diphenylchlorophosphate (1.10 mL, 0.00533 mol), diisopropylethylamine (0.927 mL, 0.00533 mol) and 4-dimethylaminopyridine (0.6 mg, 0.1 mol%).

The reaction mixture was stirred at 0 °C for 1 hour and then cooled to -20 °C and treated with a solution of 4-mercaptomethyl-2-methylpyridine (0.656 g, 0.00533 mol) in 1 ml of acetonitrile and then with 0.927 ml (0.00533 mol) of diisopropylethylamine.

The resulting mixture was stirred for 1.5 hours at -10 °C and then additional thiol (0.066 g,

0.53 mmol) and diisopropylethylamine (0.093 ml, 0.53 mmol). The reaction mixture was allowed to warm to about 10 °C over 1 hour and then diluted with 75 ml of cold ethyl acetate, washed (water and brine), dried (sodium sulfate) and evaporated (bath temperature < 30 °C).

The resulting gum was chromatographed on silica gel, eluting with ethyl acetate to remove impurities, followed by elution with acetonitrile to give the product (1.04 g, 53%) as a pale yellow foam.

¹ HNMR (CDC1 ₃ ) delta 8.43 , 7.07 (ABq, J = 5 Hz, 2H), 7.10 (s, 1H) , 6.20 to 5.75 (m, 1H) , 5.51 to 5.29 (m, 2H), 4.81 to 4.69 (m, 2H), 4.29 to 4.03 (m, 2H), 3.96 (s, 2H), 3.35 to 3.05 (m, 2H) , 2.53 (s, 3H) , 2.16 (broad s, 1H) , 1.33 (d, J = 6.3 Hz, 3H) , 1.22 (d, J = 7.3 Hz,

3H); IR (oil) 1,770, 1,705 cm” ¹ .

G. Preparation of (4R,5R,6S)-3-[[(1,2-dimethylpyridinium-4-yl)methyl]thio]-6-[i-(R)-hydroxyethyl]-4-methyl-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate

A solution of the allyl ester (0.582 g, 0.0015 mol) in 15 mL of anhydrous acetonitrile was treated under a nitrogen atmosphere with methyl trifluoromethanesulfonate (0.178 mL, 1.575 mmol) at -5 °C.

After 15 minutes, a solution of tetrakis(triphenylphosphine)palladium (0.035 g, 2 mol%) and triphenylphosphine (0.035 g) in 1 ml of methylene chloride was added, followed after 5 minutes by the addition of 0.131 ml (1.575 mmol) of pyrrolidine.

The resulting mixture was stirred for 20 minutes at 0°C and then 30 ml of cold (0°C) acetone was added. The mixture was stirred vigorously for 15 minutes at 0°C and then the precipitate was filtered off, washed with cold acetone and dried in vacuo to give 0.520 g of a beige powder.

Dilution of the filtrate with ether gave an additional 0.041 g of crude product. The combined solids were dissolved in a small amount of phosphate buffer pH 7.4 (0.05M) and applied to a reversed phase column (C10g Bondapak).

Elution with water and then 2% acetonitrile and water gave compound 17B (0.413 g, 76%) as a yellow solid after lyophilization. ¹ HNMR (D ₂ O) delta 8.55, 7.76 (ABq, J = 6.3 Hz, 2H), 7.81 (s, 1H), 4.4-3.7 (m, 2H), 4.19 (s, 2H), 4.16 (s, 3H), 3.47-3.14 (m, 2H), 2.73 (s, 3H), 1.24 (d, J = 6.4 Hz, 3H), 1.16 (d, J = 7.3 Hz, 3H); IR (KBr) 1750, 1595 cm-1 UV (phosphate buffer, pH 7.4) 293 nm (epsilon 7170).

Example 18

Preparation of (5R,6S) 3-Q.,6-dimethyl-pyridinium-2-yl)methylthio]-6-(1R-hydroxyethyl)-4R-methyl-7-oxo-1-azabicyclo[3.2.0}hept-2-ene-2-carboxylate

A. (1,6-Dimethylpyridinium-2-yl)methylthiol trifluoromethanesulfonate salt

CH ₃

About ''N'

O

SCCH,

MeOTf

-?

ether

HCl, 6N

Δ >

ch ₃

A solution of (6-methylpyridin-2-yl)methylthioacetate (1.0 g, 5.52 mmol) in anhydrous ether (5 mL) was treated under nitrogen with methyl trifluoromethanesulfonate (0.74 mL,

6.5 mmol) and stirred for 4 hours at 23 °C. The ether was decanted and the white solid was washed twice with ether (2 ml) and dissolved in hydrochloric acid solution (15 ml, 6N, 90.0 mmol)

The resulting solution was heated to 70°C for 4 hours under nitrogen and then concentrated under reduced pressure to a yellow syrup. Traces of hydrochloric acid were removed by distillation with water (2 x 10 ml). The crude material was purified by reverse phase column chromatography (2.2 x x 13.0 cm, PrepPak C-18) with water as the eluent.

The appropriate fractions were combined and lyophilized to give a white powder, 1.43 g, 85.4%, IR (KBr) λ: 2565 (SH, 1626 (pyridinium), 1585 (pyridinium cm ; UV (H_0) λ 278 inax z rnax (epsilon 7355);

Analysis: calculated for CgH ₁₂ NOgS ₂ F ₃ :

C 35.64, H 3.99, N 4.62, S 21.14;

found C 35.49, H 4.05, N 4.56, S 20.99.

B. (5R,6S) 3- (5R,6-Dimethylpyridinium-2-yl)methylthi(J-6-(1R-hydroxyethyl)-4R-methyl-7-oxo-1-azabicyclo[3,2,5]hept-2-ene-2-carboxylate

O

1-(PhO) _2PCl

2- NEt(iPr) ₂ 3- Thiol

4- Net(iPr) ₂

H ₂ ,Pd/C buffer 7.0 ether

THF

whoa

To a cold (5 C) solution of (5R,6S)-p-nitrobenzyl-3,7-dioxo-6-(1R-hydroxyethyl)-4RS-methyl-1-azabicyclo[3,2,q]heptane-2-R-carboxylate (1.11 g, 3.06 mmol, R/S: 86/14) in anhydrous acetonitrile (90 ml) under a nitrogen atmosphere, diphenylchlorophosphate (0.68 ml, 3.3 mmol) and diisopropylethylamine (0.57 ml, 3.3 mmol) were added simultaneously over 10 minutes. The cold (5 °C) mixture was stirred for 1 hour, cooled to -30 °C, and a solution of (1,6-dimethylpyridinium-2-yl)methylthiol trifluoromethanesulfonate salt (1.03 g, 3.4 mmol) in anhydrous acetonitrile (2 ml) and diisopropylethylamine (0.59 ml, 3.4 mmol) were added simultaneously over 15 minutes.

The resulting mixture was stirred for 0.5 h at -30 °C, warmed to 0 °C, diluted with cold water (35 ml) and stirred for 1 h. The resulting emulsion was poured onto the top of a reversed phase column (PrepPak C-18, 2.5 x 18 cm) and then eluted with a mixture of 25 to 50% acetonitrile in water.

Lyophilization of the appropriate fractions gave a sticky yellow solid, 1.69 g, which was dissolved in wet tetrahydrofuran (40 ml). Ether (70 ml), potassium dihydrogen phosphate-sodium hydroxide buffer (pH 7.0, 0.2 M, 50 mol) and 10% palladium on carbon (1.69 g) were added to the resulting solution and the mixture was hydrogenated for 2 hours at 290 kPa and 23 °C and then filtered through Celite.

The two phases were separated and the aqueous phase was washed with ether (2 x 20 ml) and concentrated at <23 °C under high vacuum to 15 ml and loaded onto the top of a reversed phase column (PrepPak C-18). Elution with 4% acetonitrile in water gave 0.23 g of the title compound mixed with potassium sodium diphenyl phosphate (24% by mol) after lyophilization of the appropriate fractions. Further purification on a reversed phase column (2.5 x 14 cm, PrepPak C-18) with water (400 ml) and a mixture of 10% acetonitrile in water (200 ml) as eluent gave, after lyophilization of the appropriate fractions, a yellow powder, 0.17 g 15.3 % IR (KBr) gamma _max : 1,750 (C = 0 beta-lactam), 1,625 (pyridinium), 1,600 (C = 0 carboxylate) cm ^-1 ; ¹ HNMR D ₂ ) delta: 1.12 (d, J = 7.2 Hz, CHg at C-4), 1.24 (d, J = 6.4 Hz, CH^CHOH) 2.80 (s, CH^ at C-6 pyridinium), 4.18 (CH^ at N pyridinium), 4.41 (center of AB quartet, CH^S),

7.5 to 8.4 (hydrogens on pyridinium); UV (buffer 0.05M, pH 7.0) lambda^ : 278 )epsilon 11 504)?

-λ max j^ _D -256.4 (c 0.22, H ₂ <3) ; ^{= 2} θ'θ ^h measured at 37 °C in buffer (pH 7.4) at a concentration

Example

Preparation of (5R,6S) 3-[(1,6-dimethylpyridinium-2-yl)methylthio]-6-(1R-hydroxyethyl)-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate

COO

A. Trifluoromethanesulfonate and diphenylphosphate salt of (5R,6S) p-nitrobenzyl-3-[1,6-dimethylpyridinium-2-yl)methylthio]-6-(1R-hydroxyethyl)-7-oxo-1-azabicyclo[3,2,6]hept-2-ene-2-carboxylate

MeOTf

-γ ether

1-(PhO) _2POCl

2- NEt(iPr) ₂

3- thiol

4- Net(iPr) ₂

TfO © (PhO) ₂ PO ₂ o

To a cold (5 °C) solution of (5R,6S) p-nitrobenzyl-6-(1R-hydroxyethyl)-3,7-dioxo-1-azabicyclo[2,θ]heptane-2R-carboxylate (2.14 g, 6.14 mmol) in anhydrous acetonitrile (18 mL) under nitrogen was added diphenylchlorophosphate (1.37 mL, 6.6 mmol), diisopropylethylamine (1.15 mL, 6.6 mmol) at a rate to maintain the temperature at 5 °C (7 to 10 min) and 4-dimethylaminopyridine (6 mg, 0.05 mmol).

The mixture was stirred for 1.5 hours at 5 °C and used as is. For the following procedure, this mixture is referred to as solution A. A solution of (6-methylpyridin-2-yl)methylthioacetate (1.23 g, 6.8 mmol) in anhydrous ether (10 mL) was treated with methyl trifluoromethanesulfonate (0.85 mL, 7.5 mmol) under nitrogen and stirred for 1.5 hours at 23 °C.

The ether was decanted and the white powder was washed twice with ether (2 x 10 mL) and dissolved in water (20 mL). The resulting aqueous solution was cooled to 0 °C in an oxygen-free atmosphere and treated with sodium hydroxide (4N, 3.4 mL, 13.6 mmol). The mixture was stirred for 1 h at 2 °C and then the pH was adjusted to 7.6 by addition of potassium dihydrogen phosphate.

This mixture is referred to as solution B for the following procedure. Cold (5 °C) solution A is allowed to react with solution B for 0.5 hour, while the pH is maintained between 7.25 and 7.35 by dropwise addition of 4N sodium hydroxide solution.

The mixture was stirred for 0.5 h and poured onto the top of a reversed phase column (4.0 x 18 cm, PrepPak C-18). The column was eluted with 25 to 50% acetonitrile in water. Lyophilization of the appropriate fractions gave the title compound as a yellow powder, 2.82 g (51* (PhO) ₂ PO ₂ , 49%, CF 2 SO 4 ),

80%; IR (KBr) gamma _max : 3,700-3,000 (OH), 1,772 (C = 0 beta-lactam), 1,700 (C = O ester),

625 (pyridinium), 1,590 (pyridinium), cm ³ HNMR (DMSO, d-6) delta: 1.15 (d, J = 6.2 Hz, CHjCHOH), 2.84 (s, CH^ on C-6 pyridinium), 4.16 (s, CHg and N pyridinia), 4.79 (s, SCH ₂ ),

6.6 to 7.5 f(PhO) ₂ PO ₂ J, 7.5 to 8.7 (hydrogens on pyridinium and hydrogens of pNB ester).

B. (5R,6S)3-Q(1,6-Dimethylpyridinium-2-yl)methylthio]-6-(1R-hydroxyethyl)-7-oxo-1-azabicyclo p,2,θ]hept-2-ene-2-carboxylate

COOpNB h ₂ , Pd/C

->

buffer

whoa

To a solution of the trifluoromethanesulfonate and diphenylphosphate salt (49:51) of (5R,6S) p-nitrobenzyl-3- [1,6-dimethylpyridinium-2-yl)methylthio]-6-(1R-hydroxyethyl)-7-oxo-1-azabicyclo[3,2, ojhept-2-ene-2-carboxylate, (0.87 g, 1.27 mmol) in wet tetrahydrofuran (50 ml) was added ether (50 ml), potassium dihydrogen phosphate and sodium hydroxide buffer (0.1M) (40 ml, pH 7.0) and 10% palladium on carbon (0.87 g).

The mixture was hydrogenated for 2 hours at 23°C at 248 kPa and filtered through Celite. The two phases were separated and the aqueous phase was washed with ether (2 x 15 mL), concentrated under high vacuum to 30 mL and poured onto the top of a reversed phase column (PrepPak C-18, 2.2 x 13 cm).

The column was eluted with water. The appropriate fractions were combined and lyophilized to give a yellow powder, 0.179 g, 40%: IR (KBr) .9 ^ama max ^! 1,755 (C = 0 beta-lactam), 1,628 (pyridinium),

590 (c = O carboxylate) cm“ ¹ : ¹ HNMR (D ₂ O) delta: 1.25 (d, J = 6.4 Hz, CH.jCHOH), 2.82 (s, CH ₃ at C-6 pyridinium), 3.12 ('dď, J = 9.2 Hz, J = 2.9 Hz, C-4), 3.39 (dd, J = 6.0 Hz,

J = 2.8 Hz, H-6), 3.7 to 4.4 (CHjCHOH, H-5, CH ₃ on N of pyridinium), 4.48 (s, CH ₂ S), 7.6 to 8.4 (hydrogens on pyridinium) ; UV (HO) lambda : 279 (epsilon 9 628) with inflection at 296:^]^ ¹ of max jj

55.0 °C (c 0.63, H^O) ; ⁼ ^-^.5 h measured at 37 °C in pH 7.4 buffer for a concentration of io ^{* 4} m.

Example

Preparation of 3-[2-(N-methylpyridiniummethanethiol]-6alpha-[1-(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo(3,2,0)hept-2-ene-2-carboxylate

_CO2 pNB

A. p-nitrobenzyl-2-diazo-3-oxo-n-valerate (4)

A solution of 50 g (0.35M) ethyl 3-oxo-n-valerate and 54 g (0.35M) p-nitrobenzyl alcohol in 400 ml toluene was heated at 130-140 °C for 18 hours without reflux. Evaporation of the solvent gave a yellow crystalline solid which was recrystallized from ether and pentane to give 75 g (86% yield) of p-nitrobenzyl-3-oxo-n-valerate (3), mp 33-34 °C.

IR (KBr) gamma 1,740 and 1,705 cm ^-1 . NMR (CDCl 1 ) delta 1.20 (3H, t, J = 7.0 Hz), 2.65

247195 ⁸⁴ (2H, q, J = 7.0 Hz), 3.60 (2H, s), 5.28 (2H, s), 7.45 (2H, d, J = 9.5 Hz), and 8.18 (2H, d,

J = 9.5 Hz. To a solution of 55.5 g (0.22M) of compound 3 in 500 ml of acetonitrile at 0°C was added 45 g (0.44M) of TEA and then 50 g (0.22M) of p-carboxybenzenesulfonyl azide.

The ice bath was removed and the mixture was allowed to stir for 90 minutes. The precipitate was filtered off, washed with acetonitrile and the filtrate was concentrated to about 100 ml volume and then diluted with 800 ml ethyl acetate. The organic solution was washed with aqueous sodium bicarbonate solution, brine and dried (MgSO4).

Evaporation of the solvent gives 55 g (90% yield) of compound 4 in the form of light yellow crystals, mp 96-97° C. IR (KBr) gamma 2120 and 1710 cm ^*1 .

NMR (CDCl 2 ) delta 1.20 (3H, t, J = 7.0 Hz), 2.85 (2H, q, J = 7.0 Hz), 5.40 (2H, s),

7.50 (2H, d, J = 8.0 Hz) and 8.15 (2H, d, J = 8.0 Hz).

OSi + \X>°2 ^pNB

COjpNB _ n ₂

5

B. 1-p-nitrobenzyloxycarbons1-1-diazo-2-tert.butyldimethylsilyloxy-2-butene (5)

To a cooled (0°C) solution of 54 g (0.2M) of compound 4 in 400 mL of methylene chloride was added 41.4 g (0.4M) of TEA followed by .56 g (0.21M) of tert-butyldimethylsilyl chloride in 30 mL of methylene chloride over 40 min. The solution was stirred for 120 min and then washed with ice water.

The methylene chloride was dried (MgSO4), filtered and evaporated in vacuo to give 68 g (89% yield) of compound 5^ as a yellow solid, mp 54-55°C.

IR (KBr) gamma 2,080 and 1,695 cm NMR of compound _5 shows that compound 5^ is obtained as a mixture of E/Z in the olefinic position in the ratio 9:1, NMR (CDCl2 of the major isomer) delta 0.15 (6H, s), 0.90 (9H, ε), 1.58 (3H, d, J = 7.0 Hz), 5.15 (2H, s), 7.30 (2H, d, J = 9.0 Hz) and 8.0 (2H, d, J = 9.0 Hz).

OSi + .OAc

AXIS+

-NH \xíLxCO ₂ pNB

T

->

N ₂

OSi + S ^H 3 ^N 2

X—

I ( CO ₂ pNB J—NH O

Oh, C. 4beta-1-methyl-3-diazo-3-p-nitrobenzyloxycarbonyl-2-oxo-propyl)-3alpha-[Ϊ-(R)-tert. butyldimethylsilyloxyethyl]-azetidin-2-one

To a suspension of 12.5 g (0.1 M) of anhydrous zinc chloride in 700 ml of methylene chloride was added 60.4 g (0.21 M) of compound 6, stirred for 15 minutes at 23°C and then cooled to 0°C.

A solution of 106 g (0.27M) of compound 5_ in 200 ml of methylene (

chloride and then stirred for 120 minutes without a cooling bath. The reaction mixture was washed with aqueous sodium bicarbonate solution (4 x 150 ml), water, brine and dried over magnesium sulfate.

*

Evaporation of the solvent gave a dark oil which was purified on a silica gel column. Elution of the column with a mixture of ethyl acetate and methylene chloride (1:9) gave 51.5 g (54%) of compound 1 as a white crystalline solid, mp 112-114 °C.

IR (KBr) gamma 2130, 1760 and 1720 cm h NMR at 360 MHz of compound 7_ shows that compound T_ is a mixture of methyl isomers in the 1-position in a 2:1 ratio. NMR (CDCl^) delta 0.3 to 0.6 (6H, 2s), 0.82 (9H, 2s), 1.05 to 1.15 (6H, m), 2.68 (0.88 H, q,J = 6.6 and 2.0 Hz), 2.88 (0.34 q,

J = 6.6 and 2.0 Hz), 3.57 (1H, m), 3.84 (1H, m), 4.09 (1H, m), 5.17 (2H), 5.84 (0.66 H, s), 5.95 (0.34 H, s), 7.52 (2H, d, J = 8.5 Hz), and 8.23 (2H, d, J = 8.5 Hz).

D. 4beta-(1-methyl-3-dioazo-3-p-nitrobenzyloxycarbonyl-2-oxo-propyl)-3alpha-[1-(R)-hydroxyethyl]/[beta]-azetidin-2-one (8)

To a solution of 30 g (59.5 mmol) of compound 7 in 400 ml of methanol at 23°C was added 150 ml of 1N hydrochloric acid and stirred for 18 hours. The reaction mixture was concentrated to a volume of about 200 ml and extracted with ethyl acetate (3 x 200 ml).

The combined ethyl acetate extracts were washed with water, aqueous sodium bicarbonate solution and brine. Evaporation of the solvent dried over magnesium sulfate gave 22.3 g (96%) of the title compound as a white crystalline solid, mp 147-148°C.

IR (KBr) gamma 3400, 2135 and 1750 cm ¹ , NMR at 360 MHz of compound 8 shows that compound 8 is obtained as a mixture of 4-methyl in the ratio 2:1. NMR (DMSO-d) delta 1.07 to

1.10 (6H, m), 2.75 (0.66 H, q, J = 6.6 and 2.0 Hz), 2.85 (9.34 H, J = 6.6 and 2.0 Hz), 3.55 to

3.90 (3H, m), 5.25 (2H, s), 7.70 (2H, d, J = 9.0 Hz), 8.05 (0.66H, s), 8.10 (0.34H, s), and 8.27 (2H, d, J = 9.0

Hz) .

OH

_CO2 pNB

-=?

E. p-Nitrobenzyl-6alpha-[1-(R)-hydroxyethyl]-4-methyl-3,7-dioxo-1-azabicyclo[3.2.0]hepta·-2-carboxylate (9)

A solution of 14.0 g (35.86 mmol) of compound 11 and 70 mg of rhodium(II) octanoate in ethyl acetate was heated to reflux for 20 min under nitrogen. The mixture was evaporated in vacuo to give compound 9 as a foam. IR (CHCl^) gamma 3400 and 1750 cm ³ . NMR at 360 MHz of compound 9 showed that compound 9 was obtained as a 2:1 mixture of methyls at the 4-position. The nuclear Overhauser effect was used to determine the 4-methyl configuration. When the H,. of the more abundant isomer was irradiated, there was an approximately 7% increase in the signal for the 4-methyl protons, indicating a cis configuration of H^ and 4-methyl.

Conversely, when the isomer in lesser abundance is irradiated, the desired increase in signal for 4-methyl does not occur, indicating a trans configuration of 4-methyl for the isomer in lesser abundance.

NMR (CDCl2) of the isomer with the higher delta 1.24 (3H, d, J = 7.35 Hz), 1.40 (3H, d, J = 6.3 Hz), 2.40 (1H, m), 3.24 (1H, q, J = 6.6 and 7.2 Hz), 3.76 (1H, q, J = 8.0 and 2.2 Hz), 4.18 (1H, m),

4.82 (1H, s), 5.24 (1H, d, J = 6.3 Hz), 6.18 (1H, d, J = 6.3 Hz), 7.60 (1H, d, J % 8.5 Hz), and 8.22 (1H, d, J = 8.5 Hz).

NMR (CDCl2) of the isomer with less delta 1.0 (3H, d, H = 7.35 Hz), 1.40 (3H, d,

J = 6.3 Hz), 2.83 (1H, m), 3.25 (1H, q, J = 6.6 and 1.50 Hz), 4.14 (1H, q, J = 7.36 and 1.50 Hz), 4.67 (1H, s), 5.24 (1H, d, J = 6.3 Hz), 6.18 (1H, d, J = 6.3 Hz) and 7.60 (1H, d, J = 8.5 Hz), and 8.22 (1H, d, J = 8.5 Hz).

OH CH, OH CH, ^J - -> ήπ \-OP(O0) ₂ O^ what ₂ pnb 10 1 CO ₂ pNB

F. p-Nitrobenzyl-3-diphenoxyphosphinyl-6alpha-[1-(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo (3,2,0)hept-2-ene-2-carboxylate (10)

To a cooled (0 °C) solution of 20.0 g (55.2 mmol) of keto intermediate 9^ in 150 mL of acetonitrile was added 7.18 g (55 mmol) of diisopropylethylamine followed by 14.85 g (55 mmol) of diphenylchlorophosphonate in 20 mL of acetonitrile over 5 minutes.

The resulting solution was stirred for 60 minutes at 0°C, then diluted with 600 ml of ethyl acetate, washed with ice-cold 10% phosphoric acid and brine. The mixture was dried over magnesium sulfate and evaporated to give a crude oil which was purified on a silica column.

Elution of the column with 10% ethyl acetate in methylene chloride afforded 3.7 g (11.5%) of phosphonate 10 as a white foam.

IR (CHCl-j) gamma 3400, 1790 and 1720 cm ^-1 .

NMR (CDCl ₃ ) delta 1.20 (3H, d, J = 7.2 Hz), 1.38 (3H, d, H = 7.3 Hz), 3.35 (1H, 1,

J = 6.7 and 2.0 Hz), 3.50 (1H, m), 4.2 to 4.25 (2H, m,, 5.20 (1H, d, J = 10.5 Hz), 5.37 (1H, d, J = 10.5 Hz), 7.1 to 7.4 (10H, m), 7.56 (1H, d, J = 9.0 Hz), and 8.10 (1H, d, J = 9.0 Hz).

The nuclear Overhauser effect is used to determine the 4-methyl configuration of compound 10. When irradiated with H^, there is no increase in the signal for 4-methyl, indicating a trans configuration of H^ and 4-methyl.

/Xz

G. p-nitrobenzyl-3-[pyridin-2-yl-methanethiol]6alpha[1-(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo[3,2,0]hept-2-ene-2-carboxylate (11)

To a cooled (-15°C) solution of 1.2 g (2 mmol) of phosphonate 10 in 10 ml of acetonitrile was added 390 mg (3 mmol) of diisopropylethylamine and then 370 mg (3 mmol) of 2-tercaptomethylpyridine under a nitrogen atmosphere.

The reaction mixture was stirred for 60 minutes at -15°C and then for a further 60 minutes at 0°C. The reaction mixture was diluted with ethyl acetate, washed with ice water, brine and dried (MgSO4). Evaporation of the solvents in vacuo gave a yellow oil which was purified on a silica column.

Elution of the column with 20% ethyl acetate in methylene chloride afforded 375 mg (40% yield) of compound 11 as a white amorphous foam. IR (KBr) gamma 3400, 1775 and 1710 cm \ NMR (CDCl^η delta 2.14 (3H, d, J = 6.7 Hz), 2.19 (3H, d, J = 6.7 Hz), 3.14 (1H, q, J = 6.2 and 2.0 Hz),

3.40 (1H, m), 4.0 (1H, d, J = 7.6 Hz), 4.12 (1H, d, J = 7.6 Hz), 4.18 OH, q, J = 6.7 Hz and 2.0 Hz), 4.25 (1H, m), 5.25 (1H, d, J = 11.3 Hz), 5.40 (1H, d, J = 11.3 Hz) 11.3 Hz), 7.15 to 8.2 (4H, m).

H. 3-[2-(N-methylpyridinium)methanethiol-6alpha-yl-(R)-hydroxyethyl]-4beta-methyl"7-oxo--1-azabicyclo{3,2,0)hept-2-ene-2-carboxylate (12)

To a solution of 1.0 g (2 mmol) of compound 11 in 10 ml of methylene chloride was added <50 mg (3.3 µg/mL) of methyl trifluoromethanesulfonate and stirred for 90 min at 23 °C. Evaporation of the ethyl chloride in vacuo gave the quaternized pyridine as a foam, which was hydrogenated immediately ^for further purification.

The crude pyridinium salt was dissolved in TMF-ether buffer pH 7 (1:1:1, 100 mL each) and 600 mg of 10% palladium on carbon was added. The mixture was hydrogenated for 45 minutes at 241 kPa on a shaker. The mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL).

The combined filtrate and washings were extracted with ether (2 x 100 ml) and lyophilized to give a yellow powder which was purified on a C18 Bondapak reversed phase column (10 g), eluting with 5% acetonitrile in water at 55 kPa.

Each 15 ml fraction was subjected to high-performance liquid chromatography and the fractions with ultraviolet absorption at lambda _max 300 nm were pooled and lyophilized to give 58 mg (11% yield) of the title compound as a pale yellow powder.

IR (KBr) gamma 3 400, 1 750 and 1 590 cm Lambda _max (H ₂ O) ^92 ^nm ( ^e P ^s ^l° ⁿ 7 081). NMR (D ₂ O) delta 1.13 (3H, d, J = 6.5 Hz), 1.23 (4H, d, J = 6.5 Hz), 3.18 (1H, m), 3.45 (1H, q,

J < 6.0 and 2.1 Hz), 4.0 to 4.4 (4H, m), 4.65 (3H, s), 7.79 (2H, m), 8.30 (1H, m) and 8.60 (1H, m).

Example 21

Preparation of 3-[2-(1,4-dimethylpyridinium methanethio]-6alpha-[1(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo(3,2,0)hept-2-ene-2-carboxylate

3-[2-(1,4-dimethylpyridinium-methanethio2-6alpha-[*1-(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo(3,2,0)hept-2-ene-carboxylate (13)

This compound was obtained as a yellow powder in 17% yield from compound 10 in the same manner as described in Example 20.

IR gamma 3,400, 1,755 and 1,600 cm \ UV lambda (H _o 0) 300 nm (epsilon 7,600). NMR (D _o O) max zz delta 1.20 (3H, d, J = 6.7 Hz), 1.28 (3H, d, J = 6.7 Hz), 2.60 (3H, s), 3.4 to 3.5 (2H, m), 4.2 to 4.4 (4H, m), 4.52 (3H, s), 7.82 (1H, t, J = 6.5 and 4.2 Hz), 8.32 (1H, d, J = 6.5 Hz) and 8.60 (1H, d, J = 4.2 Hz).

Example 22

Preparation of 3-[4-(1-methylpyridinium unenethi]-6alpha-[1-(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo(3,2,0)hept-2-ene-2-carboxylate

OH

1) HS

_{2 ) CF3SO3CH3}

3) Pd/H ₂ //7 coF ©

]-ch ₃

3-[4-(1-Methylpyridiniummethanethiol-6alpha-[1-(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo(3,2,0)hept-2-ene-2-carboxylate (14)

This compound was obtained as a yellow powder in 15% yield from compound 10 in the same manner as described in Example 20. IR (KBr) gamma 3 4104 1750 and 1650 cm . UV lambda _max (H ₂ 0) 293 nm (epsilon 7 295). NMR (D ₂ O) delta 1.15 (3H, d, J = 6.5 Hz), 1.20 (3H, d, J = 6.5 Hz), J = 6.5 Hz), 3.20 (1H, m), 3.45 (1H, q, J = 6.0 and 2.0 Hz), 4.11 (1H, q, J = 8.0 and 2.0 Hz),

4.20 (1H, m) and 4.35 (3H, s), 7.95 (2H, d, J = 5.2 Hz) and 8.72 (2H, d, J = 5.2 Hz).

Example 23

Preparation of 3-[3-(1-methylpyridinium)methanethio]-6alpha-[1-(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo(3,2,0)hept-2-ene-2-carboxylate

3-[3-(1-methylpyridiniummethanethio]-6alpha-Q1-(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo(3,2,0)hept-2-ene-2-carboxylate (15)

This compound is obtained as a yellow powder in 27% yield from compound 10 in the same manner as described in Example 20.

IR (KBr) gamma 3,420, 1,750 and 1,610 cm ^-1 . UV lambda _max (H ₂ O) 295 nm (epsilon 8,750).

NMR (D ₂ 0) delta 1.10 (3H, d, J = 6.9 Hz), 1.25 (3H, d, J = 6.9 Hz), 1.27 (1H, m), 1.43 (1H, q, J = 6.2 and 1.8 Hz), 4.1 to 4.35 (4H, m), 4.39 (3H, s), 8.0 (1H, t, J = 8.5 and 6.2 Hz), 8.45 (1H, d, J = 8.5 Hz), 8.70 (1H, d, J = 6.2 Hz), and 8.82 (1H, s).

Analysis: Calcd for C12H12N12O12S1/2H12O: C 51.90, H 6.36, N 7.12; Found C 51.92, H 5.71, N 6.88.

Example 24

Preparation of 3-[3-(1,2,-dimethylpyridinium)methanthic]-6alpha-[1-(R)-hydroxyethyl']-4beta-methyl-7-oxo-1-azabiocyclo(3,2,0)hept-2-ene-2-carboxylate

2) CFjSOjCHj

->

3) Pd/H ₂

3-[3-(1,2-dimethylpyridiniummethanethiol| -6alpha-[[1-(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabiocyclo(3,2,0)hept-2-ene-2-carboxylate (16)

This compound was obtained as a yellow powder in 14% yield from compound 10 in the same manner as described in Example 20.

IR (KBr) gamma 3 410, 1 750 and 1 600 cm UV lambda _max (H ₂ O) ² ^6 nm ( ^e P ^s ^-i° ⁿ ® 500).

NMR (D ₂ O) delta 1.25 (3H, d, J = 6.5 Hz), 1.30 (4H, d, J = 6.5 Hz), 2.95 (3H, s), 3.40 (1H, m), 3.50 (1H, q, J = 6.2 and 1.8 Hz), 4.2 to -4.4 (4H, m), 4.35 (3H, m). s), 7.82 (1H, t, J = 8.5 and 6.3 Hz), 8.40 (1H, d, J = 8.5 Hz) and 8.72 (1H, d, J = 6.3 Hz).

Example 25

Preparation of ^3-(2,4-dimethyl-1,2,4-triazolium)methanethioJ-6alpha-Ql-(R)-hydroxyethylj-4beta , '-methyl-1-oxo-1-azabicyclo(3,2,0)hept-2-ene-2-carboxylate ch ₃

_{2 ) CF3SO3CH3}

3) Pd/H ₂

0.3-(2,4-dimethyl-1,2,4-triazolium)methanethio]-6alpha-Q(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo(3,2,0)hept-2-ene-2-carboxylate (17)

This compound was obtained as a yellow powder in 9% yield from compound 10 in the same manner as described in Example 20.

IC (KBr) gamma 3420, 1756 and 1605 cm \ UV lambda^^ (H ₂ O) 291 nm (epsilon 7850) .

NMR (D ₂ O) delta 1.15 (3H, d, J = 6.3 Hz), 1.22 (3H, d, J = 6.3 Hz), 3.35 (ÍH, m), 3.48 (ÍH, q), J = 6.0 and 1.8 Hz), 3.90 (3H, s), 4.05 (3H, s), 4.2 to 4.4 (4H, m), 8.80 (1H, s).

Example 26

Preparation of 3-[2-(1,3-dimethylimidazolium)methanethiol]-6alpha0.-(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo(3,2,0)hept-2-ene-2-carboxylate

3-[2-(1,3-dimethylimidazoliummethanethioJ-6alpha-[1-(R)-hydroxyethyl]-4beta-methyl-7-oxo-1-azabicyclo(3,2,0)hept-2-ene-2-carboxylate (18)

This compound is obtained as a yellow powder in 32% yield from compound 10 in the same manner as described in Example 20.

iC (KBr) gamma 3400, 1758 and 1600 cm . UV lambda (H ₂ O) 294 nm (epsilon 7 194).

NMR (D ₂ O) delta 1.10 (3H, d, J = 6.3 Hz), 1.25 (3H, d, J = 6.3 Hz), 3.30 (1H, m), 3.42 (ÍH, 1, J = 6.0 and 2.2 Hz), 3.85 (6H, s), 4.2 to 4.6 (4H, m), and 7.40 (2H, s).

Example 27

By the general procedures of Examples 1 to 26, the following carbapenem products are prepared from the intermediate of formula

Example

C.

27a

-CH

27b

-ch ₂ ch ₂

N-CH

27c

-CH

CH,

27d

-CH ^CH \®

27e

-ch ₂ ch ₂ -

O® ch ₂ -ch ₂ -ch ₃

CH, \®

-p

CH,

27h

-CH _n 247195

27i

-CH _n it

CH,

I® .N.

O

CH*

27j

CHg - r---N-CH3 —CH ₃

27k

-CH _n

-<j ®r

CH,

271

CH, \ 3 -CH-O ^? ch ₃

7tn ®/

CH,

27n

-CH~27o

-CH-

CH ₃ (mixture of 3 possible

27p

Τ ^- ί Φ ' s^n-ch ₃ coo ^fc

CH,

27q

CH,

Example

C.

CH,

27 years

-CH„.©

27s

-CH _n

27t

-CH-ch ₂ -coo

CH,

I©

-AND

N—N ,© ch ₃

^.-N-CH, Ν—N

CH,

Example 28

By the general procedures of Examples 1 to 26, the following carbapenem products are prepared from the intermediate of formula

CH,

28b \©

-O

28c —^J4^ ^H 2 ^CH 2 ^CH 3

Example

C.

28d

-CH-CH, ll > ^H 3

28e

28f ^CH3 XJtc ^C H ^H 3 ³

N-

CH,

28g

CH,

-AT

28h

28i

28j

28k

281

8m i mixture j rr N—p-N CHg isomers)

CH,

N

I! © xN-CH,

COO

CH,

I — N

CH,

CH, ch ₂ -coo ch ₃

I ©

N,

N—N (mixture of 3 possible

CH,

Example

E.

About ^%s

28n

-CH

N-N-CH, >N=N

CH,

Example 29

If the keto intermediate is replaced with an equimolar amount of the corresponding 1-alpha-methyl intermediate, the above carbapenem product is obtained.

Claims (7)

OBJECT OF THE INVENTION A process for the preparation of carbapenem derivatives of the general formula I wherein θ R is hydrogen, R 6 is selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, alkenyl and alkynyl of up to 10 carbon atoms, cycloalkyl and cycloalkylalkyl of 3 to 6 carbon atoms in the alkyl moiety, phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein the heteroaryl is a mono-, di- or polycyclic radical having 5 or 6 members in individual rings and the heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic a ring having 5 or 6 members in each ring and wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 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 substituent or substituents of the aforementioned radicals are independently of one another selected from the group consisting of alkyl of 1 to 6 carbon atoms optionally substituted by amino, halogen, hydroxyl or carboxyl, halogen, —QR -OCNRV O ' - ^ ⁴ -NR ³ r ⁴ nr ^ nr ³ r ⁴ o ¹¹ ·,, -s-nr ³ r ⁴ o -NHCNR ³ r ⁴ -R ³ CNR ⁴ -CO, R ³ o -OCR ³ -SR II q -SR ^y i | 9 “NETWORK -CN'O ^: V -OS-R247195 -OP (O) (OR ³⁾ (OR ⁴⁾ 3 4 -NR CO ₂ R -N ° 2 Wherein, in the above substituents, R and R are independently selected from the group consisting of hydrogen, alkyl, alkenyl and alkynyl of up to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl of 3-6 atoms in the cycloalkyl ring and 1-6 atoms carbon in the alkyl moieties, phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms and heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl wherein heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic moiety with And 5 or 6 members in each ring and heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic radical having 5 or 6 members in each ring and wherein the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moieties linked to the heterocyclic moieties have 1 to 6 carbon atoms or R ³ and R ³ R ⁴ , taken together with the nitrogen atom to which at least one is attached, may form a 5- or 6-membered nitrogen containing heterocyclic ring, R is as defined for R in addition to hydrogen or R and R together form an alkylidene of 2 to 10 carbon atoms or alkylidene of 2 up to 10 hydroxyl-substituted carbon atoms, R is substituted or unsubstituted alkyl, alkenyl and alkynyl of up to 10 carbon atoms, cycloalkyl and cycloalkylalkyl of 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moieties, phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical having 5 or 6 members in individual rings and the heterocyclyl is mono-, di- or polycyclic a saturated or unsaturated non-aromatic heterocyclic radical having 5 or 6 members in each ring and wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with said heterocyclic moieties 6 carbon atoms, wherein the abovementioned R ³ radicals are optional substituted with 1 to 3 substituents independently selected from the group consisting of alkyl of 1 to 6 carbon atoms, optionally substituted with amino, fluorine, chlorine, carboxyl, hydroxyl or carbamoyl, fluorine, chlorine or bromine, OR ³ -OCO ₂ R ³ -OCOR ³ -contr ³ r ⁴ o AND) -OS-R 3 4 -NR CO ₂ R 3 3 4 -NR CONR R 3 »9 -NR S-R -SR Feb 9 -S-R * O \ 9 -S-R -CN phenyl optionally substituted with 1 to 3 fluorine, chlorine, bromine, alkyl of 1 to 6 carbon atoms, -OR 4, -NR ⁴ R ⁴ , -SO-R 4, -CO _? R 4 or -CONR ⁴ R ⁴ , wherein R ⁴ , R ^4, and in substituent 5, ^Z entities R are as defined above, or may be attached at another point in the ring to form a linked heterocyclic or heteroaromatic ring which may further contain further heteroatoms selected from oxygen, nitrogen and sulfur, R! 5 j _e is hydrogen, substituted and unsubstituted alkyl, alkenyl and alkynyl having up to 10 carbon atoms, cycloalkyl, cycloalkylalkyl, and alkylcycloalkyl having 3-6 carbon atoms in the cycloalkyl ring and 1 up to 6 carbon atoms in the alkyl moieties, spirocycloalkyl having 3 to 6 carbon atoms, phenyl, arylalkyl, arylalkenyl, and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl wherein heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical having 5 or 6 members in individual rings and the heterocyclyl is mono-, di- or polycyclic saturated or not a saturated 5 or 6 membered non-aromatic heterocyclic radical and wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms, and the alkyl moieties attached to these heterocyclic moieties have 1 to 6 atoms wherein the substituent or substituents of the above radicals are 100 selected from the group consisting of amino, mono-, di- and trialkylamino, hydroxyl, alkoxy, mercapto, alkylthio, phenylthio, sulfamoyl, amidino, guanidino, nitro, chloro, bromo, fluoro, cyano and carboxyl and wherein the alkyl moieties in the above substituents have 1 to 6 carbon atoms, A is a straight or branched alkylene having 1 to 6 carbon atoms, R is a hydrogen atom, an anionic charge, or a conventional carboxyl protecting group which is easily cleavable, selected from the group consisting of benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, triohloroethyl, trialkylsilyl wherein alkyls have 1 to 4 carbon atoms, phenacyl, p-methoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl, alkyl of 1 to 6 carbon atoms, pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, methoxymethyl and preferably p-nitrobenzyl, where R is also hydrogen or a protecting group is also present the ion and the group represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocyclic radical containing at least one ring nitrogen atom and a total of 1 to 4 nitrogen, oxygen or sulfur atoms, and attached A through the ring carbon atom and having a ring nitrogen atom which is quaternized with the group R ⁵ and pharmaceutically acceptable salts thereof, characterized in that the intermediate of formula II wherein r 1, R 1, _and a are as defined above, 2 ' R is a conventional, easily cleavable carboxyl protecting group selected from the group consisting of benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloromethyl, trialkylsilyl wherein alkyls have 1 to 4 carbon atoms, phenacyl, p-methoxybenzyl, acetonyl, o -nitrobenzyl, 4-pyridylmethyl, alkyl of 1 to 6 carbon atoms, pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, methoxymethyl and preferably p-nitrobenzyl, and O represents a mono-, bi- or polycyclic aromatic heterocyclic radical containing a quaternizable ring nitrogen atom, which ring is attached to A via a ring carbon atom, in an inert organic solvent with an alkylating agent of the formula X ' 101 Wherein R is as defined above and X 'is a conventional leaving group such that the R group quaternizes the nitrogen atom of the substituent ring in the intermediate of formula II to form a compound of formula I (d) where R ¹ , R ⁸ , R ¹⁵ , R ² , A, O ® RS and X 'are as defined above and optionally replace the anion X' with another anion and optionally 2 'remove the carboxyl protecting group R to form an unprotected compound of formula Or a pharmaceutically acceptable salt thereof. 1 2 2. A process for the preparation of carbapenem derivatives of the general formula I according to item 1, wherein R, R, R ° and A are as defined in point 1 and R ¹ is a hydrogen atom, characterized in that an intermediate of formula (II) wherein R, R, R, A is as defined in point 1 is reacted; and _e is hydrogen, in an inert organic solvent with an alkylating agent of the formula wherein R5 is as defined in point 1, to give a compound of formula 1 'wherein R \ R, R ^2, A, A *, and X 'are as defined in paragraph 1 and R ³⁸ is hydrogen, and optionally 2' replacing the anion X 'for another anion, and optionally removing R carboxyl protecting group to give the desired deprotected compound of formula Or a pharmaceutically acceptable salt thereof. 1 2 3. A process for the preparation of carbapenem derivatives of the general formula I according to item 1, wherein R, R, R ³ , R ⁸ and A are as defined in 1 and R ³⁸ is substituted and unsubstituted alkyl, alkenyl and alkynyl of up to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl of 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 atoms carbon in the alkyl moieties, C 3 -C 6 sirocycloalkyl, phenyl, arylalkyl, arylalkenyl and arylalkinyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein heteroaryl is mono a di- or polycyclic aromatic heterocyclic radical having 5 or 6 members in individual rings and heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic radical having 5 or 6 members in individual rings and wherein the heteroatom or heteroatoms in the aforementioned heterocyclic the moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms, and alkyl moieties The 102 attached to these heterooyl moieties have 1 to 6 carbon atoms wherein the substituent (s) of the above moieties are selected from the group consisting of amino, mono-, dia-trialkylamino, hydroxyl, alkoxy, mercapto, alkylthio, phenylthio, sulfamoyl, amidino, guanidino, nitro , chlorine, bromine, fluorine, cyano and carboxyl, and wherein the alkyl moieties in the above substituents have 1 to 6 carbon atoms, are reacted by an intermediate of formula (II) wherein R 1 is as defined in (1); ¹⁵ has the meaning given in this point. in an inert organic solvent with an alkylating agent of general formula R ⁵ -X 'wherein R' A and is as defined in point 1 to form a compound of formula I 'wherein R ¹ , R °, and X' are as defined in point 1 and is as defined in that point O* 2 'and optionally removing the carboxyl protecting group of formula (I) or a pharmaceutically acceptable salt thereof to form the desired unprotected salt compound. 4. A method according to claim 3, wherein the quaternization step is carried out after removal 2 'carboxyl protecting groups R 1 5. The process of claim 2, wherein the corresponding starting compounds of step 2 are reacted to give 3- (N-methylpyridin-2-yl-methanthio) -6alpha- [1- (R) -hydroxyethyl] -7. oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate. 6. The process of clause 3, wherein the corresponding starting compounds of clause 3 are reacted to give 3- (N-methylpyridin-2-yl-methanthio) -4alpha-methyl6alpha-Q- (R) -hydroxyethyl]. -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate. 7. The process of claim 3, wherein the corresponding starting compounds of point 3 are reacted to give 3- (N-methylpyridin-2-yl-methanthio) -4-beta-methyl-6alpha- [1- (R)]. -hydroxyethyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate.