NO843541L - PROCEDURE FOR MANUFACTURING CARBAPENEM - Google Patents

Abstract

Antibiotically active carbapenem derivatives having a 2-substituent having the formula. wherein A represents a C to alkylene group A at a ring carbon atom and quaternized with the substituent, is prepared by reacting the carbapenem core containing a conventional leaving group in the 2-position, with a thiol of the formula. the X ^ is an amine.

Description

The present invention relates to a special method for the production of carbapenem antibiotics where the 2-substituent has the formula

wherein A means a C-]-g straight chain or branched alkylene group, R 5 means an optionally substituted aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, aryl, araliphatic, heteroaryl, heteroaraliphatic, heterocyclyl or heterocyclyl-aliphatic group, and

means a nitrogen-containing, aromatic heterocyclic group, which is attached to the alkylene group A at a ring carbon atom and quaternized with the substituent R₂.

The carbapenem derivatives produced according to the method of the invention are described in US serial number 425 755 of 28 September 1982 and in a CIP application to this with the same submission date as the US application corresponding to the present application.

US serial number 425 755 and this CIP application describe the preparation of carbapenem antibiotics of the formula

wherein R 8 is hydrogen and R 1 is selected from hydrogen; substituted and unsubstituted alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl and cycloalkylalky1 with 3-6 carbon

atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic part are selected from the group comprising 1-4 oxygen, nitrogen or sulfur atoms and the alkyl parts connected to the heterocyclic parts have 1-6 carbon atoms; in which the substituent or substituents with regard to the above-mentioned groups are independently selected from C. I — o, alkyl, which is optionally substituted with amino, halogen, hydroxy or carboxyl halogen

wherein with respect to the above-mentioned substituents, the groups R 3 and R 4 are independently selected from hydrogen; alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl parts; phenyl; aralkyl, arralkenyl and aralkynyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms, and the alkyl parts in connection with these hetero-3 4

cyclic parts have 1-6 carbon atoms or R and R form together with the nitrogen atom to which at least one is bound,

a 5- or 6-membered nitrogen-containing heterocyclic ring, R<9>

is as defined for R 3 , except that it cannot be hydrogen; or wherein R 1 and R 8 together form the C 2 -Q alkylidene substituted with hydroxy, R 1 is selected from substituted and unsubstituted alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl and cycloalkylalkyl with 3-6 carbon atoms

in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the heteroatom or atoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moieties, which are connected to the heterocyclic moieties, have 1-6 carbon atoms , in which the above-mentioned R groups are optionally substituted with 1-3 substituents independently chosen from:

C 1 -C 8 alkyl, which is optionally substituted with amino, fluorine, chlorine, carboxyl, hydroxy or carbamoyl; fluorine, chlorine or bromine;

phenyl, which is optionally substituted with 1-3 fluorine, chlorine, bromine, C..-C -alkyl, -OR3, -NR3R4, -SO-R3, -CO„R3 or -CONR3R4, 6 3 4 9 5 3 2 wherein R, R and R in these R substituents are as defined

above; or R<5> may be bound to

at another site on the ring to form a fused heterocyclic or heteroaromatic ring, which ring may contain additional heteroatoms selected from O, S and N,

R 1 5 is selected from the group consisting of hydrogen, substituted and unsubstituted alkyl, alkenyl and alkynyl with from 1 to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl parts, spirocycloalkyl with 3- 6 carbon atoms, phenyl, aralkyl, aralkenyl and aralkynyl, in which the aryl part is phenyl and the aliphatic part has 1-6 carbon atoms, heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from 1-4 oxygen, nitrogen and sulfur atoms and the alkyl moieties connected to the heterocyclic moieties have 1-6 carbon atoms, in which the substituent or substituents, which relate to the above-mentioned groups, are selected from the group consisting of amino, mono-, di- and trialkylamino, hydroxyl, alkoxyl , mercapto, alkylthio, phenylthio, sulfamoyl, amidino, guanidino, nitro, chlorine, bromine, fluorine, cyano and carboxy, and wherein the alkyl parts of the above-mentioned substi tuents have 1-6 carbon atoms,

2

A is straight chain or branched alkylene, R is hydrogen, an anionic charge or a conventional easily removable carboxyl protecting group provided that, when R 2 is hydrogen or a protecting group, a counterion is also present; and

means a substituted or unsubstituted mono-, bi- or polycyclic, aromatic, heterocyclic group containing at least one nitrogen atom in the ring and bonded to A through a ring carbon atom and with a ring nitrogen, which is quaternized by the group R 5 , and pharmaceutically acceptable salts thereof, by the procedure shown in the following reaction core:

In a preferred variant of the above-mentioned method, the form is modified as shown below:

More specifically, the starting material (III) is reacted in an inert organic solvent such as methylene chloride, acetonitrile or dimethylformamide, with approx. an equimolecular amount of an agent 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 or the like, wherein L is the corresponding leaving group such as toluenesulfonyloxy, p-nitrobenzenesulfonyloxy, diphenoxyphosphinyloxy and other leaving groups, which are formed by conventional methods and which are well known in the art. Reaction to form the cleavable group in the 2-position of intermediate III) is advantageously carried out in the presence of a base such as diisopropylethylamine, triethylamine, 4-dimethylaminopyridine or the like, at a temperature between approx. -20° and +40°C, preferably at approx. 0°C. The cleavable group L in intermediate (IV) can also be halogen, in which case such a group is formed by reacting intermediate (III) with a halogenating agent such as 03PC12, 03PBr2, (00)3<p>Br2, oxalyl chloride or the like in a solvent such as Cr^C^, CH^CN, THF or the like, in the presence of a base such as diisopropylethylamine, triethylamine, 4-dimethylaminopyridine or the like. Intermediate product (IV) can, if desired, be isolated, but is suitably used for the next step without isolation or purification.

Intermediate (IV) is then converted to intermediate (II) by a conventional yield reaction. Thus, intermediate (IV) can be reacted with an equimolecular amount of a heteroaralkyl marcaptan reagent with the formula

wherein A means the C^_g straight chain or branched alkylene, and

means a mono-, bi- or polycyclic aromatic heterocyclic group containing a quaternisable nitrogen atom in the ring,

which ring may be bonded to A through a ring carbon atom, 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 yield is not critical, but an advantageous temperature interval is from approx. -40° to +25°C. The turnover is most appropriately carried out by cooling, e.g. at approx. 0°C to -10°C.

The quaternization of the ring nitrogen in the heteroaralkyl group in intermediate (II) is carried out by reacting intermediate (II) in an inert organic solvent with at least one equivalent (up to approx. 50% molar excess) of an alkylating agent of the formula

where R 5 has the meaning stated above and X<1> is a conventional leaving group such as halogen (chlorine, bromine or iodine, most preferably iodine) or a sulfonate ester moiety such as a mesylate, tosylate or triflate. Examples of suitable non-reactive organic solvents are chloroform, methylene chloride, tetrahydrofuran, dioxane, acetone, dimethylsulfoxide and dimethylformamide. The temperature for the alkylation reaction is not critical and temperatures in the range between approx. 0°C and 40°C are preferred. Most advantageously, the reaction step is carried out at room temperature.

Intermediate product (I<1>) will have in connection with it an opposite ionX<1> (e.g. derived from the alkylating agent used), which at this stage or at a later stage, i.e. after the deblocking stage, can be substituted of another opposite ion, e.g. one that is pharmaceutically acceptable, using conventional procedures. Alternatively, the counterion can then be removed during the deblocking step.

The deblocking step for removal of the carboxyl protecting group R 2 ' in the intermediate (I') is carried out by means of conventional procedures such as solvolysis, chemical reduction or hydrogenation. When a protecting group such as p-nitrobenzyl, benzyl, benzhydryl or 2-naphthylmethyl is used, which can be removed by catalytic hydrogenation, the intermediate (I<1>) in a suitable solvent such as dioxane-water-ethanol, tetrahydrofuran-aqueous dipotassium hydrogen -phosphate-isopropanol or the like, is treated under a hydrogen pressure of from 1 to 4 atmospheres in the presence of a hydrogenation catalyst such as palladium-on-charcoal, palladium hydroxide, platinum oxide or the like, at a temperature from 0 to 50 o C in from about. 0.24 to 4 hours. When R 2 ' is a group such as o-nitrobenzyl, photolysis can also be used for the deblocking. Protecting groups such as 2,2,2-trichloroethy1 can be removed by mild zinc reduction. 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 conventional carboxyl protecting groups can be removed using methods known per se. Finally, as mentioned above, compounds of formula (I<1>) where R 2 ' is a physiologically hydrolyzable ether, such as acetoxymethyl, phthalidyl, indanyl, pivaloyloxymethyl, methoxymethyl, etc., can be administered directly to the host without deblocking, as such esters are hydrolyzed in vivo under physiological conditions.

The method described above has several disadvantages. Thus, the method includes e.g. several steps, the number of which could be advantageously reduced. The overall reaction yield is also quite low and the quaternization step is carried out on the carbapenem compound itself. It would be desirable to have a new process for the preparation of compounds of formula (I) which (1) involves fewer steps, (2) gives higher yields, (3) allows the quaternized amine to be formed first and then attached to the carbapenem nucleus on a later step in the synthesis and (4) can be used for an easier formation of quaternary amine products with a large number of amines, i.e. amines with steric hindrance and those with low pKb values.

The present invention provides a special method for the production of carbapenem derivatives with

the formula

wherein R 8 is hydrogen and R 1 is selected from the group consisting of hydrogen; substituted and unsubstituted alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl and cycloalkylalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the heteroatom or atoms in the above-mentioned heterocyclic part are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl parts which are connected to the heterocyclic parts have 1-6 carbon atoms; wherein the substituent or substituents with respect to the above-mentioned groups are independently selected from the group consisting of

C1-g alkyl which is optionally substituted with amino, halogen, hydroxy or carboxyl

halogen

wherein with respect to the above-mentioned substituents, the groups R 3 and R 4 are independently selected from hydrogen; alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl1 with 3-6 carbon atoms in cycloalkyl-

the ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl parts in connection with these hetero-3 4 cyclic parts have 1- 6 carbon atoms or R and R form, together with the nitrogen atom to which at least one is attached, a 5- or 6-membered, nitrogen-containing, heterocyclic ring, R 9 is as defined for R<3> except that it cannot be hydrogen; or wherein R 1 and R 8 together form the C 2 -Q alkylidene substituted with hydroxy, R 1 is selected from the group consisting of substituted and unsubstituted alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl and cycloalkylalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the heteroatom or atoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms, and the alkyl moieties connected to the heterocyclic moieties have 1-6 carbon atoms; in which the above-mentioned R 5 groups are optionally substituted with 1-3 substituents independently selected from: C 1 -C 8 alkyl which is optionally substituted with amino, fluorine, chlorine, carboxyl, hydroxy or carbamoyl; fluorine, chlorine or bromine; phenyl, which is optionally substituted with 1-3 fluorine, chlorine, bromine, C1-C1-alkyl, -OR3, -NR3R4, -SOoR3, -COoR3 or -CONR3R4, 1 6 3 4 9 5 3 2 in which R , R and R in these R substituents are as defined above; or R may be bound to

at another point on the ring, so as to form a fused heterocyclic or heteroaromatic ring which may contain additional heteroatoms selected from O, S and N;

R 1 5 is selected from the group consisting of hydrogen, substituted and unsubstituted alkyl, alkenyl and alkynyl with 1-10 carbon atoms, cycloalkyl, cycloalkylalky1 and alkylcycloalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl parts, spirocycloalkyl with 3-6 carbon atoms, phenyl, aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has 1-6 carbon atoms, heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from the group consisting of 1- 4 oxygen, nitrogen and sulfur atoms and the alkyl moieties, which are connected to the heterocyclic moieties, have 1-6 carbon atoms, in which the substituent or substituents with respect to the above-mentioned groups are selected from the group consisting of amino, mono-, di- and trialkylamino, hydroxyl, alkoxy, mercapto, alkylthio, phenylthio, sulfamoyl, amidino, guanidino, nitrogen, chlorine, bromine, fluorine, cyano and carboxy, and wherein the alkyl moieties in the above each of said substituents has 1-6 carbon atoms,

A is C^_g straight chain or branched alkylene, R 2 is hydrogen, an anionic charge or a conventional easily removable carboxyl protecting group, provided that when R 2 is hydrogen or a protecting group, a counter ion is also present; and

means a substituted or unsubstituted mono-, bi- or polycyclic, aromatic, heterocyclic group containing at least one nitrogen atom in the ring, which ring is bonded to A through a ring carbon atom and has a ring nitrogen quaternized by the group R^, and pharmaceutically acceptable salts thereof , which method is characterized by reacting an intermediate product with the formula 18 15 2' in which R , R and R have the above meaning, R is a conventional easily cleavable carboxyl protecting group and L is a conventional cleavable group such as toluene-sulfonyloxy, p-nitrobenzenesulfonyloxy, diphenoxyphosphinyloxy or halogen, with a thiol compound of the formula wherein A and has the above meaning and #<>> is an opposite anion, in an inert solvent and in the presence of a base to form the carbapenem compound of the formula 8 15 in which R, R, R, R ,A,

and has the above meaning, and if desired removes the carboxyl protecting group R 2 ' to form the corresponding deblocked compound of formula (I) or a pharmaceutically acceptable salt thereof.

The present invention also provides intermediates of formula (VII).

The carbapenem compounds of formula (I) are powerful antibacterial agents or intermediates which can be used for the preparation of such agents.

The compounds of formula (I) above contain the carbapenem nucleus and can therefore be called 1-carba-2-penem-3-carboxylic acid derivatives. Alternatively, the compounds can be considered with the base structure

and are designated as 7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylic acid derivatives. Although the present invention includes compounds in which the relative stereochemistry of the 5,6-protons is both cis and trans, the preferred compounds have 5R,6S-(trans)-stereochemistry as is the case with thienamycin.

The compounds of formula (I) may be unsubstituted

in the 6-position or substituted with substituent groups previously described for other carbapenem derivatives. More particularly, R 8 can be hydrogen and R 1 can be hydrogen or a non-hydrogen substituent referred to e.g. in European application

8 1 No. 38,869 (see the definition of Rg). Alternatively, R and R can together form the C 2 -6 alkylidene with e.g. hydroxy substituted C 1 -Q alkylidene. The compounds of formula (I) can also be unsubstituted in the 1-position (R^<5>=H) or substituted with substituent groups previously described in connection with other carbapenem derivatives. More particularly, R 1 5 can be hydrogen or any of the non-hydrogen 1 substituents such as e.g. is described in European application no. 54,917 (see defini-1 2

the sion for R or R herein) or US-PS 4,350,631. Preferred non-hydrogen R 15 substituents include C 1-6 alkyl,

most preferably methyl, phenyl and phenyl(C 1 if )alkyl. The non-hydrogen R 15 substituent can either be in a or B configuration, and it is intended that the present invention includes both the individual a and B isomers and mixtures thereof.

1 8 15

The definitions of R , R , and R are further explained below: (a) The aliphatic "alkyl", "alkenyl" and "alkynyl" groups may be straight-chain or branched with 1-10 carbon atoms, preferably 1-6 and most preferably 1- 4 carbon atoms; when they form part of another substituent such as in cycloalkylalkyl or heteroaralkyl or aralkenyl, the alkyl, alkenyl and alkynyl group preferably contains 1-6 and preferably 1-4 carbon atoms. (b) "Heteroaryl" includes mono-, bi- and polycyclic aromatic heterocyclic groups containing 1-4 O, N or S atoms; preferred are 5- or 6-membered heterocyclic rings such as thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, tetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, pyrazolyl, etc. (c ) "heterocyclyl" includes mono-, bi- and polycyclic, saturated or unsaturated, non-aromatic heterocyclic groups containing 1-4 O, N or S atoms; preferred are 5- or 6-membered heterocyclic rings such as morpholinyl, piperazinyl, piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrrolinyl, pyrrolidinyl, etc. (d) "Halogen" includes chlorine, bromine, fluorine and iodine and is preferably chlorine , fluorine or bromine.

The term "conventional easily removable carboxyl protecting group" refers to a known ether group which has been used to block a carboxyl group during the chemical reaction steps described below, and which can be removed if desired by methods which do not result in any significant destruction of the remainder of the molecule, e.g. by chemical or enzymatic hydrolysis, treatment with chemical reducing agents under mild conditions, irradiation with ultraviolet light or catalytic hydrogenation. Examples of such ester protecting groups include benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloroethyl, silyl such as trimethylsilyl, phenacyl, p-methoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl and C 1-6 alkyl , such as methyl, ethyl or t-butyl. Such protecting groups include those that are hydrolyzed under physiological conditions such as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethyl. A particularly advantageous carboxyl protecting group is p-nitrobenzyl which can be easily removed by catalytic hydrogenolysis.

The above-mentioned pharmaceutically acceptable salts include the non-toxic acid addition salts, e.g. salts with mineral acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid, etc., 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 malic acid. Compounds of formula (I) in the form of acid addition salts can be written as

(R<2>= H or protecting group) where X means the acid anion. The opposite anion X may be chosen so as to provide pharmaceutically acceptable salts for therapeutic administration, but in the case of intermediates of formula (I) X may also be a toxic anion. In such a case, the ion can then be removed or substituted with a pharmaceutically acceptable anion to form an active end product for therapeutic use. When acidic or basic groups are present in the R^ or R 5 group or on

group, the present invention also includes suitable base or acid salts of these functional groups, e.g. acid addition salts in the case of a basic group and metal salts (e.g. sodium, potassium, calcium and aluminium), the ammonium salt or salts with non-toxic amines (e.g. trialkylamines, procaine, dibenzylamine, 1-e -phenamine, N-benzyl-8-phenethylamine, N,N<1->dibenzylethylenediamine, etc.) in the case of an acidic group.

Compounds of formula (I) in which R 2 is hydrogen, an anionic charge or a physiologically hydrolyzable ester group are, together with pharmaceutically acceptable salts thereof, useful as antibacterial agents. The other compounds of formula (I) are valuable intermediates that can be converted into the biologically active compounds mentioned above.

/ n preferred embodiment of the present invention comprises compounds of formula (I) in which R Q is hydrogen and

1

R is hydrogen, CE^ CH -,

Within this subgroup, the preferred compounds are those where R 1 is

OH

in

CH-jCH- and the most preferred compounds have the absolute configuration 5R, 6S, 8R.

Another preferred embodiment comprises compounds of formula (I) in which R 1 and R 8 together form an alkylidene group with the formula

The alkylene group (ie substituent "A") in the compounds of formula (I) can be straight-chain or branched and contain from 1-6 carbon atoms. A preferred embodiment if-

includes such compounds in which A is -(CH_)- , where n is 1

2. n

or 2 and a particularly preferred embodiment comprises such compounds where A is -CH,,-.

The alkylene part "A" is via a ring carbon atom bound to

an N-substituted, quaternized, aromatic, heterocyclic group of the general formula

in which the R^-substituent is preferably an optionally substituted Cj-Cg-alkyl, C^-C^Q-alkenyl, C2-Cio"alkynyl'C3-C6-cycloalkyl, C^-Cg-cycloalkyl-Cj-Cg-alkyl, phenyl, phenyl-Cj-Cg-alkyl, phenyl-C^-Cg-alkenyl, phenyl-C2-Cg-alkynyl, heteroaryl, heteroaralkyl, in which the alkyl moiety has 1-6 carbon atoms. The heteroaryl substituent (or heteroaryl moiety of heteroaralkyl)

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

The R 5 substituent can optionally be substituted with 1-3 substituents independently selected from: (a) C,I -Crb-alkyl which is optionally substituted with preferably 1-3 amino-, fluorine-, chlorine-, carboxyl-, hydroxy- or carbamoyl groups; (b) fluorine, chlorine or bromine;

(u) phenyl which is optionally substituted with 1-3 substituents independently selected from fluorine, chlorine, bromine, C^-Cg-alkyl, -OR 3,

-NR3R4 4, -SO^R<3>, -CO R<3>or -CONR3R54, in which the groups R<3> and R in relation to the above-mentioned R -substituents are independently selected from hydrogen, alkyl, alkenyl and alkynyl with 1-10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl parts; phenyl, aralkyl, aralkenyl and aralkynyl, in which the aryl part is phenyl and the aliphatic part has 1-6 carbon atoms and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the heteroaryl and heterocyclyl group

or - part of a group as defined above for R 1 , and the alkyl moieties associated with the heterocyclic moieties have 1-6 carbon atoms; or R 3 and R 4 form together with the nitrogen atom to which at least one is bound, a 5- or 6-membered, nitrogen-containing, heterocyclic (as defined above 5 9 3 for R ) ring; and R has the meaning given above for R except that it cannot be hydrogen. A particularly preferred R -substituent 5 is C 1 -C 6 -alkyl and especially methyl. Moreover, the R -substituent can together with another ring atom of the moiety

form a fused heterocyclic or heteroaromatic ring which may contain further, preferably 1 or 2, heteroatoms selected from 0, N and S. For example,

preferably represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocyclic ring containing at least one nitrogen atom in the ring and 0-5 further ring heteroatoms selected from 0, S and N, the heterocyclic ring being bound to A through a ring carbon atom and has a ring nitrogen atom quaternized by the group R^.

The heteroaromatic ring

may optionally be substituted at available ring carbon atoms with preferably 1-5, and preferably 1-3 substituents independently selected from C^_^-alkyl, C^_^-alkyl substituted with preferably 1-3 hydroxy, amino, _^- alkylamino, di(C^_^)-alkylamino, Cj _4~ alkoxy, carboxy, halogen (which shall hereafter mean chlorine, bromine, fluorine or iodine, preferably chlorine, bromine or fluorine) or sulfo, C^g-cycloalkyl, C -^g-cycloalkyl(C^_4)alkyl, which is optionally substituted with 1-3 substituents mentioned above in connection with C^_^-alkyl, C1_4- alkoxy, _4-alkylthio, amino, _^-alkylamino, di(C^_^)-alkylamino, halogen, _^-alkanoylamino, C _4~alkanoyloxy, carboxy, sulfo,

hydroxy,

amidino, guanidino, phenyl, phenyl substituted with 1-3 substituents independently selected from among amino, halogen, hydroxy, trifluoromethyl, C^_4~alkyl, _4~alkyl, _4~alkylamino, di(C^_4)alkylamino, carboxy, sulfur, phenyl(Cj _4)alkyl, in which the phenyl part can optionally be substituted with 1-3 substituents mentioned above in connection with phenyl and the alkyl part can optionally be substituted with 1-3 substituents mentioned above in connection with C^_^-alkyl, and heteroaryl or heteroaralkyl, in which the heteroatom or atoms are selected from the group consisting of 1-4 O-, S- or N-atoms and the alkyl part connected to the heteroaralkyl has 1-6 carbon atoms, the heteroaryl and heteroaralkyl groups in the heterocyclic ring part being optionally substituted with 1-3 substituents independently selected from hydroxy, amino, halogen, trifluoromethyl, <C>1_4-alkyl, _4~ alkoxy, C1_4~ alkylamino, di(C^ _4) alkylamino, carboxy and sulfo and in the alkyl part with 1-3 substituents selected among hydroxy, amino, _4~alkylamino, di(C^_4)alkylami no, _4~Alkoxy, carboxy, halogen and sulfo. Moreover, available ring nitrogen atoms (not the quaternized nitrogen) can be substituted with 1-3 substituents independently selected from C^_4~alkyl, C^^-alkyl substituted with preferably 1-3 hydroxy, amino, _4~alkylamino,. di (C ^ _4) alkylamino , Cj _4~ alkoxy, carboxy, halogen or sulfo groups, C^_g-cycloalkyl, C3_g-cycloalkyl(Cj _4)alkyl, which is optionally sub-

substituted with 1-3 substituents mentioned above in connection with C^_4~alkyl, phenyl, phenyl substituted with 1-3 substituents independently selected from among amino, halogen, hydroxy, trifluoromethyl, C^^-alkyl, _4~alkoxy, Cj _4- alkylamino, di-(Cj _^)alkylamino, carboxy and sulfo, phenyl(C^alkyl, in which the phenyl part may optionally be substituted with 1-3 substituents mentioned above in connection with phenyl and the alkyl part may optionally be substituted with 1-3 substituents mentioned above in connection with C1-4~alkyl, and heteroaryl or heteroaralkyl, in which the heteroatom or atoms are selected from 1-4 0-, S- or N-atoms and the alkyl part which is connected to the heteroaralkyl has 1-6 carbon atoms, the heteroaryl being - and the heteroaralkyl groups in the heterocyclic ring part are optionally substituted with 1-3 substituents independently selected from hydroxy, amino, halogen, trifluoromethyl, C^_^-alkyl, _4~ alkoxy, _4-alkylamino, di(C^_4)alkylamino, carboxy and sulfo and in the alkyl part with 1-3 substituents selected from hydroxy, amino, Cj _4~alkylamino, di(C^_^)alkylamino, Cj _4~alkoxy, carboxy, halogen and sulfo. The most preferred ring carbon atom and the most preferred nitrogen substituents are C 1-6 alkyl, especially methyl.

Within the preferred embodiment described above, the preferred compounds are those where A is -(CH2 0) n, where n is 1 or 2, most preferably those where A is -CH_-18

and where (a) R and R together mean

or (b) when R 8 is hydrogen and R 1 means hydrogen, CH^Ct^-,

Particularly preferred are the compounds where R g is hydrogen and <1>

R is

especially compounds with the absolute configuration

5R, 6S, 8R.

In a preferred embodiment, the group means

an aromatic 5- or 6-membered, N-containing heterocyclic ring containing 0-3 additional heteroatoms selected from 0, S or N. Such aromatic heterocyclic rings may, where possible, be fused to another ring which may be a saturated or unsaturated carbocyclic ring, preferably a C^_^-carbocyclic ring, an aromatic carbocyclic ring, preferably a phenyl ring, a 4-7 membered heterocyclic ring (saturated or unsaturated) containing 1-3 heteroatoms selected from 0, 11 11 S, N or NR where R is hydrogen, C^_g-alkyl which is optionally substituted with 1-2 substituents independently chosen 3 3 4 3 from -OR , -NR R , -CO-R , oxo, phenyl, fluorine, chlorine, bromine, 3 3 4 -S0-.R and -C0NR R , or phenyl optionally substituted with 1-3 3 3 4 substituents independently selected from C1 ,-alkyl, -OR , -NR R , 3 3 3 4 3 fl4 uor, chlorine,1b1rom , -SO., R , -CO., R and -CONR R , wherein R and R in such R -substituents have the meanings given above in connection with substituent R 1 , or a 5-6-membered heteroarom atic ring containing 1-3 heteroatoms and selected from 0, S, N or NR where R 11 has the meaning stated above. The 5- or 6-membered aromatic quaternized ring or, where appropriate, the fused carbocyclic, heterocyclic or heteroaromatic ring or both such rings may optionally be substituted on available ring atoms with preferably up to a total of 5 substituents for the overall ring system, of the above in connection with the group

said substituents.

Within the preferred embodiment described above, the preferred compounds are those where A is ~(CH2^n'where n is 1 or 2, most preferably those where A is -CH9-18

and (a) R and R together mean

or (b) when R 8 is hydrogen and R 1 is hydrogen, CH^Cr^-, Particularly preferred are the compounds where R is hydrogen and<1>R is

especially connections with the absolute con-

figuration 5R, 6S, 8R.

A further preferred embodiment of the invention comprises compounds of formula (I), where

means a group selected from

where R^, R^ and R^ are independently selected from hydrogen, C^_^-alkyl, C^_4~alkyl substituted with preferably 1-3 hydroxy, C1_4~alkylamino, di(C^_4~alkyl)amino, _4 ~Alkoxy, amino, sulfo, carboxy or halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C-^g-cycloalkyl, C^_^-

alkoxy, Cj _4~alkylthio, amino, _4~alkylamino, di(C^ _4~alkyl)-amino, halogen, (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), _4-alkanoylamino, _4~alkanoyloxy , carboxy,

hydroxy, amidino, guanidino, phenyl, phenyl substituted with one, two or three amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyl, trifluoromethyl, C^_4~alkyl- or _4~alkoxy groups , phenyl(C^_^)-alkyl, in which the phenyl part can optionally be substituted with 1-3 substituents mentioned above in connection with phenyl and the alkyl part can optionally be substituted with 1-3 substituents mentioned above in connection with C^_4~alkyl, and heteroaryl and heteroaralkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic part are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part connected to the heteroaralkyl part has 1-6 carbon atoms, or in which two of R^, R^ or R^ together can be a fused, saturated carbocyclic ring, a fused aromatic carbocyclic ring, a fused non-aromatic heterocyclic ring or a fused heteroaromatic ring, the fused rings being optionally substituted with 1 or 2 of the above for R^, R^ and R^ defined substituents,

optionally substituted on a carbon atom with one to three substituents independently selected from C^_4~alkyl, Cj^-alkyl substituted with preferably 1-3 hydroxy, _^-alkylamino, sul-

fo, di (C 1-4 -alkyl) amino, _4-alkyl, amino, carboxy or halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C-1-6-cycloalkyl, _4-methoxy, C 2 -alkylthio, amino, _4~alkylamino, di(Cj _4~alkyl)amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C ..-alkanoylamino,

Cj _4~alkanoyloxy, carboxy,

hydroxy, amidino, guanidino, phenyl, phenyl substituted with one, two or three amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyl, trifluoromethyl, C 1-4 -alkyl or -4-alkyl groups, phenyl(C^_^)-alkyl, in which the phenyl part may optionally be substituted with 1-3 substituents mentioned above in connection with phenyl and the alkyl part may optionally be substituted with 1-3 substituents mentioned above in connection with Cj_4-alkyl, and heteroaryl or heteroaralkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part connected to the heteroaralkyl part has 1-6 carbon atoms, or optionally substituted to form a fused carbocyclic, heterocyclic or heteroaromatic ring which is optionally substituted with 1 or 2 of the above defined substituents,

optionally substituted on a carbon atom with one or two substituents

tuents independently selected from C^_4-alkyl, C^^-alkyl substituted with preferably 1-3 hydroxy, _4~alkylamino, sulfo, di(Cj _4~alkyl)amino, _4~ alkoxy, amino, carboxy or halogen (chloro, bromine, fluorine, or iodine, preferably chlorine, fluorine, or bromine), C1-6-cycloalkyl, _4~ alkoxy, C1_4-alkylthio, amino, _4alkylamino, di(C^_^-alkyl)amino, halogen (chlorine, bromine , fluorine or iodine, preferably chlorine, fluorine or bromine), C1-4-alkanoylamino,

_4~alkanoyloxy, carboxy,

hydroxy, amidino, guanidino, phenyl, phenyl substituted with one, two or three amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyl, trifluoromethyl, C^_^-alkyl- or Cj _4 ~Alkoxy groups, phenyl(Cj_^)-alkyl, in which the phenyl part may optionally be substituted with 1-3 substituents mentioned above in connection with phenyl and the alkyl part may optionally be substituted with 1-3 substituents mentioned above in connection with C^^-alkyl , and heteroaryl or heteroaralkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part connected to the heteroaralkyl part has 1-6 carbon atoms, or optionally substituted to form a fused carbocyclic , heterocyclic or heteroaromatic ring, which is optionally substituted with one or two of the above defined substituents, optionally substituted on a carbon atom with a substituent independently selected from C^_4~alkyl, C^_4-a alkyl substituted with preferably 1-3 hydroxy, C^_4~alkylamino, di(C^_4~alkyl)-amino, sulfo, _4~alkoxy, amino, carboxy or halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C-1.g-cycloalkyl, _4- alkoxy, _4-alkylthio , amino, C.j _4-alkylamino , di (C-1_4-alkyl) amino , halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C^_4~alkanoylamino, _4-alkanoyloxy, carboxy, hydroxy, amidino, guanidino, phenyl, phenyl substituted with one, two or three amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine) , hydroxyl, trifluoromethyl, C^_4~alkyl-or _4~alkoxy groups, phenyl(C^_4)-alkyl, in which the phenyl part may optionally be substituted with 1-3 substituents mentioned above in connection with phenyl and the alkyl part may optionally be substituted with 1 -3 substituents mentioned above in connection with C^_4~alkyl, and heteroaryl or heteroaralkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic parts are selected among 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part connected to the heteroaralkyl part has 1-6 carbon atoms,

wherein X is 0, S or NR, where R is C^-alkyl, C1-4-alkyl substituted with 1-3 hydroxy, amino, Cj 4-alkylamino, di(C^-4)-alkylamino, <_>4- alkoxy , carboxy, halogen or sulfo groups,

C^_g-cycloalkyl, g-cycloalkyl(C^_4)alkyl optionally substituted with 1-3 substituents mentioned above in connection with Cj_4~alkyl, phenyl, phenyl substituted with 1-3 substituents which are independently selected from amino, halogen, hydroxy , trifluoromethyl, C^_4~alkyl, Cj _4~alkyl, Cj _4-alkylamino, di(Cj _^)alkylamino, carboxy and sulfo, phenyl(Cj _4)alkyl, which in the phenyl part may optionally be substituted with 1-3 substituents mentioned above in connection with the phenyl and the alkyl part may optionally be substituted with 1-3 substituents mentioned above in connection with C^_4-alkyl, and heteroaryl and heteroaralkyl, in which the heteroatom or atoms are selected from 1-4 0-, S- or N atoms and the heteroaralkyl-linked alkyl part have 1-6 carbon atoms, the heteroaryl and heteroaralkyl groups being optionally substituted in the heterocyclic ring part with 1-3 substituents independently chosen from hydroxy, amino, halogen, trifluoromethyl, C^_4~alkyl, _4 ~Alkoxy, Cj _4~alkylamino, di(C^_4)alkylamino, carboxy and sul fo and in the alkyl part with 1-3 substituents chosen from hydroxy, amino, Cj _4~alkylamino, di(C^_^)-alkylamino, _4~ alkoxy, carboxy, halogen and sulfo, the heteroaromatic group being optionally substituted on a carbon atom with one or more substituents independently selected from C1_4-alkyl, C1-4-alkyl substituted with preferably 1-3 hydroxy, amino, C1_4-alkylamino, di(C1_4-alkyl)amino, _4-alkyl, sulfo, carboxy or halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C1-4-cycloalkyl, C1-4-alkyl, C1-4-alkylthio, amino, -4-alkylamino, di(C^_^-alkyl) amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C._.-alkanoylamino, C._.-alkanoyloxy, carboxy,

hydroxy, amidino, guanidino, phenyl phenyl

substituted with one, two or three amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyl, trifluoromethyl, C^^-alkyl- or Cj_4~ alkoxy groups, phenyl(C^_4)- alkyl, in which the phenyl part can optionally be substituted with 1-3 substituents mentioned above in connection with phenyl and the alkyl part can optionally be substituted

tuated with 1-3 substituents mentioned above in connection with C1_4-alkyl, and heteroaryl or heteroaralkyl in which the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part connected to the heteroaralkyl part has 1-6 carbon atoms, or optionally substituted so that a fused carbocyclic, heterocyclic or heteroaromatic ring is formed, optionally substituted with 1 or 2 of the above

defined substituents,

wherein X is 0, S or NR, where R is C 1 -alkyl, C 3 -alkyl substituted with 1-3 hydroxy, amino, _4-alkylamino, di-(C 1 _4)-alkylamino, 3-alkyl, carboxy, halogen or sulfo groups, C3_g-cycloalkyl, C3_6-cycloalkyl(C^_4)alkyl optionally substituted with 1-3 substituents mentioned above in connection with C^-alkyl, phenyl, phenyl substituted with 1-3 substituents independently selected from amino, halogen, hydroxy, trifluoromethyl, C₁-alkyl, C₁-alkyl, C₁-alkylamino, di(C₄₄)alkylamino, carboxy and sulfo, phenyl(C₄₄)-alkyl, which in the phenyl part may optionally be substituted with 1-3 substituents mentioned above in connection with the phenyl and the alkyl part may optionally be substituted with 1-3 substituents mentioned above in connection with C^_4~alkyl, and hetero-

aryl and heteroaralkyl in which the heteroatom or atoms are selected from the group consisting of 1-4 O-, S- or N-atoms and the alkyl part connected to the heteroaralkyl has 1-6 carbon atoms, the heteroaryl and heteroaralkyl groups being optionally substituted in the heterocyclic ring part with 1-3 substituents independently selected from hydroxy, amino, halogen, trifluoromethyl, C^_4~alkyl, Cj _4~ alkoxy, _4~alkylamino, di-(Cj _4)alkylamino, carboxy and sulfo and in the alkyl part with 1-3 substituents selected from hydroxy, amino, _^-alkylamino, di(Cj_^)alkylamino, _4- alkoxy, carboxy, halogen and sulfo, the heteroaromatic group being optionally substituted on a carbon atom with a substituent selected from C^ ^-alkyl, C^_4~alkyl substituted with preferably 1-3 hydroxy, amino, _4~alkylamino, di(Cj_4~alkyl)amino, Cj_4~ alkoxy, sulfo, carboxy or halogen (chlorine, bromine, fluorine or iodine , preferably chlorine, fluorine or bromine), C3_g-cycloalkyl, C^__-alkyloxy, _4~alkylthio , amino, _^-alkylamino, di (C^^-alk l) amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C. _4~alkanoylamino, _4~alkanoyl-

oxy, carboxy,

hydroxy, amidino, guanidino, phenyl, phenyl substituted with one, two or three amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyl, trifluoromethyl, C^^-alkyl- or C^ ^-Alkoxy groups, phenyl(Cj _^)-alkyl, in which the phenyl part may optionally be substituted with 1-3 substituents mentioned above in connection with phenyl and the alkyl part may optionally be substituted with 1-3 substituents mentioned above in connection with C.j_4- alkyl, and heteroaryl or heteroaralkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part connected to the heteroaralkyl part has 1-6 carbon atoms, and in which R is C1-4-alkyl substituted with 1-3 hydroxy, amino, C1 _4~alkylamino, di(C^_4)-alkylamino, _4~ alkoxy, carboxy, halogen or sulfo groups, C3_g-cycloalkyl, C^g-cycloalkyl-(C^_4)alkyl optionally substituted with 1-3 substituents mentioned above in connection with C^^-alkyl, phenyl, phenyl substituted with 1-3 substituents independently selected from amino, halogen, hydroxy, trifluoromethyl, Cj_4-alkyl, Cj _4~ alkoxy, Cj _4~ alkylamino, di(C^_4)alkylamino, carboxy and sulfo, phenyl(C^_4)alkyl, which in the phenyl part can optionally be substituted with 1-3 substituents mentioned above in connection with phenyl and the alkyl part can optionally be substituted with 1-3 substituents mentioned above in connection with C^_4~alkyl . substituted in the heterocyclic ring part with 1-3 substituents independently selected from hydroxy, amino, ha logene, trifluoromethyl, C^-alkyl, C_4~ alkoxy , C1_4-alkylamino, di(C^_4)alkylamino, carboxy and sulfo and in the alkyl part with 1-3 substituents selected from hydroxy, amino, C^_4~alkylamino , di(C 1 -4 ._ )-alkylamino, C 1 -4,- alkoxy, carboxy, halogen and sulfo. The R and R groups can also together form a fused heterocyclic or heteroaromatic ring.

Within the preferred embodiment described above, the preferred compounds are those where A is~(CH2^n'in which n is 1 or 2, most preferred are those where A is

1 8

-Cr^- and (a) R and R together mean

or (b) R 8 is hydrogen and R<1> is hydrogen, CH^CH,,-,

Particularly preferred are the compounds where R g is hydrogen and <1> is

R is

especially compounds with the absolute configuration

5R, 6S, 8R.

A particularly preferred embodiment of the invention comprises compounds of formula (I), where

means a group with the formula

where R , R and R are independently selected from hydrogen, C 1 -alkyl-5 alkyl, C 1-4 -alkyl, carboxyl and carbamoyl and R has the above meaning, and is preferably C 1-6 -alkyl, most preferably

-CH3.

Within the preferred embodiment described above, the preferred compounds are those where A is -(CH2 n) n, where n is 1 or 2, most preferably those where A is -CH -

18

and where (a) R and R together mean

or (b) R 8 is hydrogen and R<1> is hydrogen, CH^Cf^-,

Particularly preferred are the compounds where R 8 is hydrogen and R<1> is OH

CH-jCH-, especially compounds with the absolute configuration 5R, 6S, 8R.

Another preferred embodiment of the invention comprises compounds of formula (I), wherein

means a group with the formula where R^ is C^_4-alkyl, most preferably methyl, and R^ means hydrogen or C^_4-alkyl, 5 6 7 where R is C^-alkyl, most preferably methyl and R and R are hydrogen or C 4 -alkyl,

where R is C^_4-alkyl, most preferably methyl and R is C^_^-alkyl or phenyl(Cj_^)alkyl, 5 6 in which R is C^^-alkyl, most preferably methyl and R is C1-4-alkyl, preferably methyl,

wherein R^ is C^_4-alkyl, most preferably methyl and R is C^_^-alkyl, most preferably methyl, or

where R<5> is C 1-4 alkyl, preferably methyl.

Within the preferred embodiment described above, the preferred compounds are those where A is -(CH2„) n, where n is 1 or 2, most preferably those where A is -CH9-18

and where (a) R and R together mean

or (b) R 8 is hydrogen and R<1> is hydrogen, CE^ CE^- i

Particularly preferred are the compounds where R g is hydrogen and R<1> is OH

CH-jCH-, especially compounds with the absolute configuration 5R, 6S, 8R.

A most preferred embodiment of the invention comprises compounds of formula (I), wherein

means a group with the formula

Within the preferred embodiment described above, the preferred compounds are those where A is -(CH2 n) n, where n is 1 or 2, most preferably those where A is -CH_-18

and where (a) R and R together mean

or (b) R 8 is hydrogen and R<1> means hydrogen, CH^CH,,-, Particularly preferred are the compounds where R g is hydrogen and R<1> is

especially compounds with the absolute con-

figuration 5R, 6S, 8R.

Particularly preferred compounds according to the invention are those with the formula

wherein R 2 is hydrogen, an anionic charge, or a conventional, readily removable carboxyl protecting group provided that, when R 2 is hydrogen or a protecting group, a counterion is also present, wherein<1>HNMR(D20)- the spectrum shows characteristic peaks at 1.23 (3H, d, J=6.4 Hz), 3.12 (2H, q, J=1.4, 8.9 Hz), 3.39(1H, q, J =2.7, 6.0 Hz), 4.07-4.68 (10H, m), 8.19 (1H, s), in which the 1HNMR(D20) spectrum shows characteristic peaks at 6: 1.23 ( 3H, d, J=6.4 Hz), 3.15 (2H, q, J=3.7, 9.0 Hz), 3.37 (1H, q, J=2.6, 6.0 Hz ), 3.95-4.65 (10H, ra), 8.62 (1H, s), A most preferred embodiment of the invention comprises compounds of formula (I), in which

Within the preferred embodiment described above, the preferred compounds are those where A is -(CH2„) n, where n is 1 or 2, most preferably those where A is -CH_-18

and where (a) R and R together mean

8 1 or (b) when R is hydrogen and R is hydrogen, CH^Cf^-, Particularly preferred are compounds where R gives hydrogen and <1>R is

especially connections with the absolute con-

figuration 5R, 6S, 8R.

The method according to the invention uses the intermediate products with the formula

like for example. is mentioned in the European application no. 38,869 and the European application no. 54,917 and which can be produced using the general methods described there. L means a conventional leaving group (defined as "X" in European Application No. 38,869) such as chlorine, bromine, iodine, benzenesulfonyloxy, p-toluenesulfonyloxy, p-nitrobenzenesulfonyloxy; methanesulfonyloxy, trifluoromethanesulfonyloxy, dephenoxy-phosphinyloxy or di(trichloroethoxy)phosphinyloxy. The preferred leaving group is diphenoxyphosphinyloxy. Intermediates of formula (IV) are usually formed in situ by reacting an intermediate of the formula 18 15 2' where R f R , R and R have the meanings given above, with a suitable acylating agent R 1 -L. The preferred intermediate (IV) where L is diphenoxyphosphinyloxy can be prepared by reacting keto-ester III in an inert organic solvent such as methylene chloride, acetonitrile or dimethylformamide, with approx. 1 equimolecular amount of diphenylchlorophosphate in the presence of a base such as diisopropylethylamine, triethylamine, 4-dimethylaminopyridine or the like at a temperature between approx. -20°C and +40°C, preferably at approx. 0°C. Intermediate product (IV) can be isolated if desired, but can suitably be used as starting material for the method according to the invention without isolation or purification. In the method according to the invention, the carbapene intermediate (IV) is reacted with a quaternary aminothiol compound with the formula in which

has the above meaning and

X

<©>

is an opposite anion.

The reaction is carried out in an inert solvent such as acetonitrile, acetonitrile-dimethylformamide, tetrahydrofuran, tetrahydrofuran-f 2 O, acetonitrile-H 2 O or acetone in the presence of a base. The nature of the base is not significant. Examples of suitable bases are sodium hydroxide, diisopropylethylamine, 1,8-diazabicyclo^5.4.o7undec-7-ene, 1, 5-diazabicyclo/4.3.07non-5-ene and tri(C^_^)alkylamines such as triethylamine, tributyl -amine or tripropylamine. Reaction of intermediate (IV) and thiol (VII) can be carried out within a large temperature range, e.g. -15°C up to room temperature, but preferably carried out at a temperature between approx. -15 and +15°C, preferably at approx. 0°C.

The carbapenem product produced by reaction of the quaternary amine thiol (VII) with intermediate (IV) will be associated with an opposite anion (e.g. C6H50)2P02® , Cl® or the anion associated with the quaternary thiol), as in this step may be substituted with another opposite anion, e.g. one that is pharmaceutically more acceptable, by conventional methods.

Alternatively, the counter anion may be removed during the subsequent deblocking. If the quaternized carbapenem compound and the counter anion form an insoluble product, the product can be crystallized as it forms and collected cleanly by filtration.

After formation of the desired carbapenem product, the carboxyl-protecting group R 2 'of 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, intermediate (I<1>) in a suitable solvent such as dioxane-water-ethanol, tetrahydrofuran- diethyl ether buffer, tetrahydrofuran-aqueous dipotassium hydrogen phosphate-isopropanol or the like, is treated under a hydrogen pressure of between 1 and 4 atmospheres in the presence of a hydrogenation catalyst such as palladium-on-charcoal, palladium hydroxide, platinum oxide or the like at a temperature between 0 and 50°C in between approx. 0.24 to 4 hours. When R is a group such as nitrobenzyl, photolysis can also be used for deblocking. Protecting groups such as 2,2,2-trichloroethyl can also be removed by mild zinc reduction. 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. In a similar way, other conventional carboxyl protecting groups can be removed using methods known to those skilled in the art. Finally, compounds of formula (I') where R 2 1 is a physiologically hydrolyzable ester such as acetoxymethyl, phthalidyl, indanyl, pivaloyloxymethyl, methoxymethyl, etc., as mentioned above, can be administered directly to the host after deblocking, as such esters are hydrolyzed in in vivo under physiological conditions.

18 5 15

It will be clear that when the R -, R -, R - or R - substituent or the quaternized heteroaromatic group attached to the substituent A contains a functional group that can interfere with the planned course of reaction, such a group can be protected by a conventional blocking -group and then unblocked to restore the desired functional group. Suitable blocking groups and methods for introducing and removing such groups are well known to those skilled in the art.

As is the case with other 3-lactam antibiotics, compounds of the general formula (I) can be converted by known methods into pharmaceutically acceptable salts, which for the purposes of the present invention are essentially equivalent to the non-salt-formed compounds. Thus, one can e.g. dissolving a compound of formula (I) wherein R 2 is an anionic charge, in a suitable inert solvent and then adding one equivalent of a pharmaceutically acceptable acid. The desired acid addition salt can be recovered by conventional methods, e.g. solution precipitation, lyophilization etc. If there are other basic or acidic functional groups present in the compound of formula I, pharmaceutically acceptable base addition salts and acid addition salts can be prepared in a similar manner using known methods.

A compound of formula (I) where R 2 is hydrogen or an anionic charge, or a pharmaceutically acceptable salt thereof, can also be converted by conventional methods into a corresponding compound where R 2 is a physiologically hydrolyzable ester group, or a compound of formula (I ) where R 2 is a conventional carboxyl protecting group, can be converted to the corresponding compound where R 2 is hydrogen, an anionic charge or a physiologically hydrolyzable ester group, or a pharmaceutically acceptable salt thereof.

It will be understood that certain products within the scope of formula (I) can be formed as optical isomers as well as epimeric mixtures thereof. It is intended that the present invention should encompass all such optical isomers and epimeric mixtures. For example, when the 6-substituent is hydroxyethyl, such a substituent can either be in R or S configuration, and the resulting isomers and epimeric mixtures thereof are covered by the present invention.

The thiol intermediates of formula (VII) can e.g. is prepared from the corresponding thioacetate compound with the formula

where A has the above meaning and means a mono-, bi- or polycyclic aromatic heterocyclic group containing a quaternisable nitrogen atom in the ring, the ring being bound to A through a ring carbon atom. The thio-acetate compound is quaternized by reacting it in an inert organic solvent such as diethyl ether, dichloromethane, methylene chloride, dioxane, benzene, xylene, toluene or mixtures thereof, with an appropriate alkylating agent of the formula

in which R^ has the meaning given above and X<1> is a conventional leaving group such as halogen (chloro, bromine or iodine, preferably iodine) or a sulfonate ester moiety such as mesylate, tosylate or triflate. The temperature in the alkylation reactor is not critical and temperatures in the area approx. 0°C

until 4C<T>C is withdrawn.

Before the reaction with carbapene intermediate (IV), the quaternized thioacetate compound is subjected to acid or base hydrolysis to form the quaternary thiol intermediate (VII). This hydrolysis is preferably carried out before coupling with (IV) to reduce decomposition of the relatively unstable, quaternary thiol (VII).

With the right choice of solvents, the reaction from intermediate product (III) to final product (I) can be carried out without isolation of the various intermediate products, i.e. in a "one-flask" process. An example of such a process is illustrated below in example 7.

The carbapenem derivatives of the general formula (I)

where R 2 is hydrogen, an anionic charge or a physiologically hydrolyzable carboxyl protecting group, or the pharmaceutically acceptable salts thereof, are powerful antibiotics which are active against various gram-positive and gram-negative bacteria, and they can be used e.g. as animal food additives, as preservatives in foodstuffs,

as bactericides in industrial applications, e.g. water-based paint and in the waste water from<p>apir mills to inhibit the growth of harmful bacteria and as disinfectants to destroy or inhibit the growth of harmful bacteria on medical and dental equipment. However, they are particularly valuable for the treatment of infectious diseases in humans and animals caused by gram-positive or gram-negative bacteria.

The pharmaceutically active compounds in question can be used alone or formulated as pharmaceutical preparations which, in addition to the active carbapenem component, comprise a pharmaceutically acceptable carrier or an excipient. The connections can be managed using a variety of means; those of primary interest include: oral, topical or parenteral (intra-venous or intramuscular injection). The pharmaceutical preparations can be used in solid form such as capsules, tablets, powders, etc., or in liquid form as solutions, suspensions or emulsions. Injection preparations, which are the preferred route of administration, can be prepared in unit dose form in ampoules or in multi-dose containers and can contain formulation agents, such as suspension, stabilization and dispersing agents. The preparations may be in a ready-to-use form or in powder form for reconstitution at the time of administration with a suitable vehicle such as sterile water.

The dose to be administered depends to a large extent

of the particular compound used, the particular preparation formulated, the mode of administration, the nature and condition of the host and the particular situs and organism treated. Selection of the particularly preferred dose and route of administration is therefore left to the therapist. Generally, however, the compounds can be administered parenterally or orally to mammalian hosts in an amount from about 5 to 200 mg/kg/day. Administration is usually carried out in divided doses, e.g. three to four times daily.

The following examples shall illustrate the invention in more detail without limiting it.

Example_el_1_

Preparation of 3-/ 4-( N, N- dimethyl- 1, 2, 3- triazolium)- methylthio/- 6ot-/ T-( R) - hydroxyethyl7- 7- oxo- 1 - azabicyclo ( 3. 2 . 0 ) hept-2-ene-2-carboxylate

Methyl trifluoromethanesulfonate (0.58 mL, 5.16 mmol) was added dropwise to an ice-cooled stirred slurry of 4-(methanethiolacetate)-1-methyl-1,2,3-triazole (590 mg, 3.52 mmol )

in 2 ml of dry methylene chloride under nitrogen. After 3 hours the bath was removed and after 1 hour the solvent was removed with a suction device. The remaining oil was dissolved in a few ml of water and this solution was cooled in an ice bath. A cold solution of sodium hydroxide (305 mg, 7.59 mmol) in a few mL of water was then added and the reaction mixture was stirred for 3/4 hour. The solution was diluted to 25 ml of water and the pH adjusted to 7.5 by adding solid sodium dihydrogen phosphate monohydrate. Then 14 ml of this solution (approx. 1.9 mmol of the triazolium thiol) was added to an ice-cooled and stirred solution of the enol phosphate (1.0 g, 1.72 mmol) in 10 ml of tetrahydrofuran (THF). This solution was stirred for 3/4 hour (some crystalline material, probably Na 2 HPO 4 , was deposited during this reaction). The suspension was transferred to a pressure bottle using 20 ml of THF and 20 ml of water. Then 30 ml of ether and 1.0 g of 10% palladium-on-charcoal were added and the mixture was hydrogenated under 40 P.S.I. for 1 hour. The organic phase was separated and washed with 2 x 5 ml of water. The combined aqueous phases were filtered and the filtrate concentrated under high vacuum (about 0.5 mm, 1} hour). The yellow solution was chromatographed (intermediate pressure reverse phase column, 35 x 90 mm, ^0 as eluent), whereby after lyophilization 395 mg of carbape-

which was somewhat contaminated with some inorganic material. It was purified by HPLC (10 x 300 mm Waters Micobondapack C-18 column, multiple injections, H 2 O as eluent) for

obtaining 310 mg (57%) of isomer A as an amber powder: <1>HNMR (D2O) 6 : 1.23 (3H, d, J=6.4 Hz), 3.10 (2H, d, J =9.1 Hz), 3.24 (1H, q, 3=2, 1, 6.1 Hz), 4.03-4.71 (10H, m), 8.46 (1H, s); IR (nujol) 1760 cm"<1>; uv (phosphate buffer, pH 7.4, M=0.05)^max<:>296 (t=7,500).

B. Preparation of Isomer B and Isomer C

Methyl trifluoromethanesulfonate (1.60 mL, 14.0 mmol) was added dropwise to an ice-cooled solution of 4-(methanethiolacetate)-2-methyl-1,2,3-triazole (1.20 g, 7.02 mmol) in 6 ml of dry methylene chloride under nitrogen. This mixture was allowed to warm to room temperature and then stirred for 16 hours. Additional methyl trifluoromethanesulfonate (0.40 ml, 3.56 mmol) was added and after 3 hours at room temperature the solvent was removed with suction. The remaining oil was triturated with ether and the resulting gum dissolved in 5 ml of water. This solution was cooled in an ice bath and a solution of sodium hydroxide (844 mg, 21.1 mmol) in 5 mL of water was added. After stirring for 3/4 hour, this solution was diluted to 60 ml with water and the pH adjusted to 8 by adding solid potassium dihydrogen phosphate. Then 40 ml of this solution (ca. 4.7 mmol of a mixture of isomeric triazolium thiols) was added to an ice-cooled, stirred solution of the enol phosphate (2.00 g, 3.45 mmol) in 60 ml of THF. This solution was stirred in an ice bath in ? hour after which it was transferred to a pressure bottle containing a suspension of 2 g of 10% palladium-on-charcoal and 60 ml of ether. The mixture was hydrogenated under 40 P.S.I. for 1 hour. The organic phase was separated and washed with 2 x 10 ml of water. The combined aqueous phases were filtered and the filtrate concentrated under a high vacuum (about 0.5 mm, 1 hour). The remaining solution was then chromatographed (intermediate pressure reverse phase column, 45 x 130 mm, H 2 O as eluent) whereby after lyophilization 595 mg of a mixture of isomeric carbapenems contaminated with some inorganic material were obtained. This mixture was separated and purified by HPLC (10 x 300 mm Waters Microbondapack C-18 column, multiple injections, H 2 O as eluent) to give in sequence: 153 mg (13%);<1>HNMR (D 2 O) 6: 1 .23 (3H, d, J=6.4 Hz), 3.12 (2H, q, J=1.4, 8.9 Hz), 3.39 (1H, q, J=2.7, 6 .0 Hz), 4.07-4.68 (10H, m), 8.19 (1H, s); IR (nujol) 1755 cm"<1>; uv (phosphate buffer, pH 7.4, M=0.05) *max: 296 nm (t=6,700), and isomer C; 284 mg (24%); 1HNMR ( D20) 6: 1.23 (3H, d, J=6.4 Hz), 3.15 (2H, q, J=3.7, 9.0 Hz), 3.37 (1H, q, J= 2.6, 6.0 Hz), 3.95-4.65 (10H, m), 8.62 (1H, s); IR (nujol) 1750 cm"<1>; uv (phosphate buffer, pH 7.4, M=0,<05>)<X>max: 298 nm (t=7,600).

Eczema<p>_<e>l_2

( 5R, 6S)- 6-( 1R- hydroxyethyl- 3-( 2- methyl- 1, 2, 3- thiadiazolium- 4-ylmethylthio)- 7- oxo- 1- azabicyclo/ 3. 2. 07hept- 2- one - 2- carboxylate

A. Ethyl-1,2,3-thiadiazol-4-ylcarboxylate1

iCD. Hurd and R.I. Mori, J. Am. Chem. Soc. , 5349(1955).

A solution of ethyl α-N-carbethoxyhydrazone propionate (31.2 g, 0.154 mol) in 80 mL of thionyl chloride was stirred at 23°C for 3 hours and heated to 70°C for 20 minutes. Thionyl chloride was evaporated and the residue triturated in 4 x 30 ml of hexane. The red solid was dissolved in 150 ml of dichloromethane and the solution washed with saturated sodium bicarbonate solution and water. After drying over Na 2 SO 4 , the solution was concentrated until crystallization. After standing at 23°C for some time, the crystals were filtered, and 16.8 g with a melting point of 86°C were obtained, the yield corresponding to 69%. The filtrate was concentrated and purified by chromatography on a silica gel column with dichloromethane as elution solvent to give 3.17 g, mp 86°C, 33%, ir (KBr)^max: 1720 (ester) cm ; Hmr (CDCl 3 ) 6 : 1.52 (3H, t, J=7.1 Hz, CH 3 CH 2 O), 4.57 (2H, q, J=7.1 Hz, CH 3 CH 2 O), 9.47 (1H, s, H of thiadiazole).

B. 1,2,3-thiadiazol-4-yl-methanol1

To a suspension of ethyl 1,2,3-thiadiazole-4-y1-carboxylate (18.35 g, 0.116 mol) in 400 ml of ether was added portionwise lithium aluminum hydride (2.47 g, 0.065 mol) during of 1 hour. The reaction mixture was stirred at 23°C for 7 h and treated with lithium aluminum hydride (2.47 g, 0.065 mL). Stirring was continued for 24 hours before 7 ml of water, 7 ml of 15% sodium hydroxide solution and 21 ml of water were added in sequence. After stirring for 15 minutes, the ether solution was decanted and the gum extracted with 5 x 100 ml of ether. The ether extracts were combined, dried over (MgSO 4 ) and concentrated. 5.4 g were obtained. The crude material was purified on a silica gel column (120 g, 4 x 16 cm) with ether as elution solvent to give 1.3 g (7%) of ethyl 1,2,3- thiadiazol-4-yl carboxylate and 2.45 g (18%) of 1,2,3-thiadiazol-4-yl methanol; ir (film) vmax: 3380 (OH) cm<-1>;<1>Hmr (CDC13)<5: 2.31 (1H, s, OH), 5.22 (2H, s, CH2O), 8, 50 (1H, s, H of thiadiazole).

<1>S.I. Råmsby, S.O. Øgren, S.B. Ross and N.E. Stjern-strøm, Acta Pharm. Succi., 10, 285-96 (1973); C. A., 79, 137052W (1973).

C. 1,2,3-thiadiazol-4-yl-methanol-methanesulfonate

A solution of 1,2,3-thiadiazol-4-yl-methanol (0.75 g, 6.5 mmol) in dichloromethane (20 mL) was cooled to 5°C under a nitrogen atmosphere and treated with triethylamine (1.018 mL, 7.3 mmol) and methanesulfonyl chloride (0.565 mL, 7.3 mmol). After 15 minutes, the ice bath was removed and the reaction mixture was stirred for 2 hours. The solution was washed with 2 x 2 mL 1N hydrochloric acid solution and water, dried over MgSO 4 + MgO and concentrated. The residue was purified by chromatography (silica gel column 1.5 x 21 cm) with ether as elution solvent to give 0.90 g (71%) of 1,2,3-thiadiazol-4-yl-methanol-methanesulfonate;

ir (film)v, : 3350 (SO_)cm"<1>;<1>Hmr (CDC1,)6: 3.09 (3H, s,

rrici.KSz.jt

CH 3 ), 5.75 (2H, s, CH 2 ), 8.72 (1H, s, H of thiadiazole); uv (CH 2 Cl 2 ) Amakg: 251 (e1 990). Analysis: calculated for CgHgN^S: C 24.73, H 3.11, N 14.42, S 33.02; found: C 24.78, H 3.09,

N 14.66, S 31.94 and 0.13 g (19%) di(1,2,3-thiadiazol-4-yl-methyl) ether; ir (film)%ax:<1>272, 1242, 1200 , 986 , 805, 728 cm"1; 1Hmr (CDC13)6: 5.16 (s, 4H, CH2), 8.42 (s, 2H , H's of thiadiazole).

D. 4- acetylthiomethyl- 1, 2, 3- thiadiazole

To a solution of 1,2,3-thiadiazol-4-yl-methanol methanesulfonate (0.90 g, 4.6 mmol) in tetrahydrofuran (9 ml) was added 2 ml of an aqueous solution of sodium thiol acetate (prepared from thiolacetic acid (0.38 mL, 5.3 mmol) and sodium bicarbonate (0.445 g, 5.3 mmol)). The resulting mixture was stirred at 23°C for 1 hour and diluted with 75 mL of ether. The organic solution was washed with 3 x 23 ml of water, dried over MgSO 4 and concentrated. The crude mixture was purified by chromatography (silica gel column: 1.4 x 19 cm) with 50% ether in hexane as eluent to give 0.60 g (75%); ir (fil<m>)<vm>ax: 1675 (C=0) cm"<1>;<1>Hmr (CDC13)6: 2.37 (3H, s, CH3), 4.58 (2H, s, CH2>, 8.44 (1H, s, H of thiadiazole). Analysis: calculated for C5H6N2OS2: C 34.47, H 3.47,

N 16.08, S 36.80; found: C 34.48, H 3.83, N 16.28, S 36.80.

E. 4- acetylthiomethyl- 2- methyl- 1, 2, 3- thiadiazolium trifluoromethanesulfonate and 4- acetylthiomethyl- 3- methyl- 1, 2, 3- thiadiazolium trifluoromethanesulfonate

To a solution of 4-acetylthiomethyl-1,2,3-thiadiazole (0.60 g, 3.44 mmol) in a mixture of 4 ml of ether and 0.4 ml of dichloromethane was added a few crystals of the title compounds and trifluoromethanesulfonate (0.407 mL, 3.6 mmol) over 5 min. The reaction mixture was stirred at 23°C under a nitrogen atmosphere for 6 hours. The white solid which was a mixture of the two title compounds was filtered and washed -1 L with ether, 1.05 g, 90%: ir (KBr)v ros. , Ks: 1675 (C=0) cm ; Hmr (DMSO, d-6)6: 2.43 (3H, s, CH3COS), 3.33 (s, CH3on N-3), 4.57 (s, CH3on N-2), 4.66 (2H , s, CH2), 9.55 (H on thiadiazolium N-2), 9.66 (H on thiadiazolium N-3). Analysis: calculated for C7H9N204S3F3: C 20.27, H 2.38, N 9.45, S 32.46; found: C 24.61, H 2.57, N 8.47, S 28.21. F. 4- mercaptomethyl- 2- methyl- 1, 2, 3- thiadiazolium trifluoromethane-sulfonate and 4- mercaptomethyl- 3- methyl- 1, 2, 3- thiadiazolium- ) trifluoromethanesulfonate

A solution of a mixture of 4-acetylthiomethyl-2-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate and 4-acetylthiomethyl-3-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate (1.05 g, 3.1 mmol) in 10 ml of 6N hydrochloric acid was heated to 65°C under a nitrogen atmosphere for 1 3/4 hours. The solvent was evaporated under reduced pressure to give 0.91 g of a yellow syrup. This compound was used in the next step without purification.

G. ( 5H, 6S)- 6-( iR- hydroxyethyl)- 3-( 2- methyl- 1, 2, 3- thiadiazolium-4- yl- methylthio)- 7- oxo- 1- azabicyclo/ 3. 2. 07hept-2-ene-2-carboxylate

A cold (5 C) solution of (5R,6S)-paranitrobenzyl-6-(1R-hydroxyethyl)-3-(diphenylphosphono)-7-oxo-1-azabicyclo-(3.2.0)hept-2 -ene-2-carboxylate (1.7 g, 2.92 mmol) in tetrahydrofuran (10 mL) was treated with a solution of an impure mixture of 4-mercaptomethyl-2-methyl-1,2,3- Thiadiazolium trifluoromethanesulfonate and 4-mercaptomethyl-3-methyl-1,2,3-thiadiazoliumtrifluoromethanesulfonate (0.9 g) in a mixture of phosphate buffer (pH 7.2, 0.3M, 15 ml) and 5 ml of tetrahydrofuran. The reaction mixture was stirred for 1 hour and the pH was maintained at 7.2 with 2N sodium hydroxide solution. Stirring was continued for a further 1 hour before 50 ml of ether and 1 g of 10% palladium-on-charcoal were added. The resulting mixture was hydrogenated at 23°C under 45 psi for 2 hours and filtered through a Celite pad. The organic phase was separated, diluted with 50 ml of ether and phosphate buffer (pH 7.2, 0.3M, 20 ml) and hydrogenated with 2 g of 10% palladium-on-charcoal for 2 hours under 50 psi. The aqueous phases were combined (from the first and second hydrogenolysis), washed with ether and purified by chromatography on PrePak 500-C/18 with water as the eluent solvent to give 0.22 g of crude material. It was purified by HPLC with water as elution solvent and 0.040 g (4%) of the title compound was obtained before lyophilization. ir (KBr)v , : 3400 (br, OH), 1745 (C=0 of B-lactam), 1580 (carboxylate) cm , Hmr (D2O)6: 1.23 (3H, d, J=6.3 Hz, CH3CHOH), 3.04, 4.05, 3.16 (2H, m, H-4), 3.38 (1H, dd, J=2.8 Hz, J = 6.0 Hz, H- 6), 3.9-4.6 (2H, m, H-5, CH3<IH0H) , 4.51, 4.53, (2"s", SCH_), 4.61 (s, N<+ >CH,); uv Ho0) X . : 224 (e4345),

262 (e4980), 296 (e6885), /α/J<3>18° (c, 0.18, HO); 1^=9.8 hours (measured at a concentration of 10 in phosphate buffer pH

7.4 at 36.8°C.

Example_el_3

Potassium- 3-/ 5-( 1- carboxylatemethyl- 3- methyl- 1, 2, 3- triazolium)-methanethio7- 6a-/ T-( R)- hydroxyethyl7~ 7- oxo- 1- azabicyclo/ 3. 2. 0j - hept-2-ene-2-carboxylate

Lithium aluminum hydride (2.83 g, 70.9 mmol) was added in small portions to a stirred suspension of 1-methyl-1,2,3-triazole-4-carboxylic acid<1> (9.00 g, 70.9 mmol ) in 200 mL dry THF. The mixture was stirred at room temperature for 15 hours, after which 20 ml of a 20% aqueous solution of sodium hydroxide was carefully added over approx. 1 ml<1>s amounts. The resulting granular suspension was filtered and the solid washed with an additional 5 x 75 mL of THF. The combined THF solutions were dried over MgSO 4 and the solvent removed. The remaining yellow oil was flash chromatographed on a silica gel column (90 x 35 mm) (100 ml portions of hexane, mixtures of ethyl acetate-hexane (1:1) and (1:3) and finally ethyl acetate-methanol (9:1) as eluent). This gave 4-hydroxymethyl-1-methyl-1,2,3-triazole (3.18 g, 40%) which is a colorless oil: 1 H NMR (CDCl 3 ) 6 : 4.07 (3H, s), 4, 73 (2H, d), 7.52 (1H, s); IR (pure) 3320 cm-1.

Methanesulfonyl chloride (3.82 mL, 49.6 mmol) was added dropwise to an ice-cooled, stirred solution of the alcohol (4.67 mL, 41.3 mmol) and triethylamine (7.47 mL, 53.7 mmol) in 20 mL methylene chloride. After | hour, the solvent was removed and the remaining solid was taken up in 30 ml of acetonitrile. Potassium thiolacetate (7.06 g, 62.0 mmol) was then added and the suspension stirred for an additional 16 h. The dark-colored suspension was then concentrated and 10 ml of water was added. This mixture was extracted with 5 x 40 ml of methylene chloride. The combined extracts were dried over MgSO 4 and the solvent removed. The remaining oil was flash chromatographed on a silica gel column (90 x 36 mm) (hexane followed by a mixture of hexane:ethyl acetate (1:1) was used as eluent). This gave 4-(methanethiolacetate)-1-methyl-1,2,3-triazole (5.95 g, 84%) as a pale pink-colored solid:<1>HNMR (CDCl-J 62.40 (3H , s), 4.10 (3H, s), 4.20 (2H, s), 7.53

-1

(1H, s); IR (nujol mull) 1675 cm.

A solution of the triazole (1.00 g, 5.85 mmol) and ethyl bromoacetate (1.48 mL, 13.3 mmol) in 10 mL of dry acetonitrile was heated to 60 °C for 90 h under nitrogen. Resolution-<1>C. Pederson, Acta. Chem. Scand., 1959, 13, 888.

the agent was removed and the residual oil triturated with 4 x 25 ml of ether to give 1-methyl-3-(ethylcarboxymethyl)-4-methanethiolacetate-1,2,3-triazolium bromide as a brownish gum which was used directly.

A cold solution of KOH (0.66 g, 12 mmol) in 5 mL of water was added to an ice-cooled, stirred solution of triazolium bromide in 20 mL of water. After 20 minutes, the solution was diluted to 35 ml and sufficient solid potassium dihydrogen phosphate was added to bring the pH of the solution to 8.0. This solution was then added to a stirred, ice-cooled solution of the enol phosphate in 35 mL of THF. After } hour, the mixture was transferred to a pressure bottle containing 35 ml of ether and 1.5 g of 10% palladium-on-charcoal. It was hydrogenated at 40 psi for 55 minutes. The organic phase was separated and washed with 2 x 5 ml of water. The combined aqueous phases were filtered and the filtrate concentrated under high vacuum. The remaining material was chromatographed on a reverse phase column (35 x 120 mm) with water as eluent. Lyophilization of the carbapenem-containing fractions gave 1.20 g of a green colored solid. This was rechromatographed on a Waters Prep. 500 HPLC (PrepPak-500/C1g column) with 2% acetonitrile-water as eluent. The carbepenem-containing fractions were combined and lyophilized. This material was then rechromatographed again by HPLC (10 x 300 mm Waters Microbondapack C-18 column) with water as eluent, whereby after lyophilization the pure title compound (190 mg, 17%) was obtained as a pale yellow solid:<1> HNMR (D20) 6: 1.24 (3H, d, J=6.4 Hz), 3.07 (2H, d, J=9Hz), 3.38 (1H, q, J=2.7, 6 .0 Hz), 4.02-4.30 (3H, m), 4.29 (3H, s), 5.23 (2H, s), 8.52 (1H, s); IR (nujol mull) 1750 cm"<1>; UV (phosphate buffer, pH 7.4) X . 296 nm (e = 7,520) .

max

Example 4

Potassium- 3-/ 4-( 1- carboxylatemethyl- 3- methyl- 1, 2, 3- triazolium)-methanethio7- 6a-/ T-( R)- hydroxyethyl7- 7- oxo- 1- azabicyclo/ 3. 2. 0/hept-2-ene carboxylate

A mixture of ethyl azidoacetate (30.0 g, 0.23 mol) and propargylic acid (14.3 mL, 0.23 mol) in toluene (75 mL) was stirred at room temperature. The reaction mixture continued to be mildly exothermic for 1 hour after which it quickly became strongly exothermic and required cooling with an ice bath. After the expiration of this exothermic phase, the reaction mixture was heated under reflux for \ hour. After cooling in an ice bath, the crystalline material was collected by filtration and washed with some toluene. The impure material thus obtained (33.3 g, 72%) consisted of a single isomer ( iHNMR (DMSO-■dg) 6: 1.20 (3H, t, J=7 Hz), 4, 15 (2H, q, J=7 Hz), 5.42 (2H, s), 8.67 (1H, 3)), presumably 1-(ethylcarboxymethyl)-1,2,3-triazole-4-carboxylic acid analogue previously performed work 1.

A solution of the carboxylic acid (5.00 g, 25.1 mmol) and triethylamine (3.68 mL, 26.4 mmol) in 50 mL of dry methylene chloride was added to an ice-cooled, stirred solution of ethyl chloroformate (2.52 mL , 26.4 mmol) in 50 ml of dry methylene chloride. This purple colored solution was stirred for 12 hours after which it was washed with 10 ml of water, dried over MgSO 4 and the solvent removed. The crude mixed anhydride was dissolved in 50 mL THF and then slowly added to an ice-cooled suspension of sodium borohydride (0.72 g, 18.9 mmol) in 50 mL THF. After stirring for 1 hour, additional sodium borohydride (0.30 g, 7.9 mmol) was added and the reaction mixture remained in the ice bath for 1 hour. Then 5 ml of water was added and after 10 minutes this was followed by 3 ml of 10% aqueous HC1. After gas evolution had finished, 2 g of solid potassium carbonate were added while stirring. The organic phase was removed and the remaining white paste extracted with additional THF. The combined organic phases were dried over MgSO 4 and the solvent removed. Flash column chromatography on silica gel, eluting with hexane, mixtures of ethyl acetate-hexane and finally ethyl acetate gave 1-(ethylcarboxymethyl)-4-hydroxymethyl-1,2,3-triazole (2.04 g, 44%) as a crystalline solid: 1 HNMR (CDCl 3 ) 6 : 1.28 (3H, t, 3=1 Hz), 4.23 (2H, q, 3=1 Hz), 4.75 (2H, s), 4 .85 (2H, s), 7.73 (1H, s) .

Diisopropylazodicarboxylate (4.11 mL, 20.8 mmol) was added dropwise to an ice-cooled solution of triphenylphos-<1>C. Pederson, Acta. Chem. Scand., 1959, 13, 888.

fine (5.47 g, 20.8 mmol) in 100 mL dry THF under nitrogen. After 1 hour, an ice-cooled solution of the alcohol (1.93 g, 10.4 mmol) and thiolacetic acid (1.49 mL, 20.8 mmol) in 50 mL of dry THF under nitrogen was added to this mixture. It remained for 2 hours in the ice bath and was then allowed to stand for a further 12 hours at room temperature after which the solvent was removed. The reaction mixture was flash chromatographed on silica gel (40 g, eluting with 100 ml portions of hexane, 5%, 10%, 15%... 50% ethyl acetate-hexane). Thiolacetate-containing fractions were combined and chromatographed again on 60 g of silica gel (elution with 200 ml portions of hexane, 5%, 10%, 15%, 20% ethyl acetate-hexane and 22.5, 25, 27.5 ... 35% ethyl acetate-hexane). This gave 1.24 g (45%) of 1-(ethylcarboxymethyl)-4-methanethiolacetate-1,2,4-triazole as a crystalline solid (<1>HNMR 6: 1.28 (3H, t, J=7 Hz), 2.37 (3H, s), 3.87 (2H, s), 3.90 (2H, q, J=7 Hz), 5.12 (2H, s), 7.63 (1H, s); IR (nujol mull), 1735, 1780 cm"<1>) and a further 1.40 g of material contaminated with triphenylphosphine oxide.

Methyl trifluoromethanesulfonate (0.51 mL, 4.53 mmol) was added dropwise to an ice-cooled and stirred solution of the triazole (1.00 g, 4.12 mmol) in 5 mL of dry methylene chloride. The bath was removed after 1 hour and after a further 2 hours the solvent was removed with a suction device-vacuum. This left a white solid which was suspended in 15 ml of water and this stirred mixture was cooled in an ice bath. A solution of KOH (0.69 g, 12.4 mmol) in 5 mL of water was added and the reaction mixture stirred for 1 hour. It was then diluted to 30 ml with water and solid potassium dihydrogen phosphate was added to bring the pH to 8.0. A portion of this solution (22 mL, ca. 3.0 mmol thiol carboxylate) was added to an ice-cooled stirring solution of the enol phosphate (1.60 g, 2.76 mmol) in 30 mL THF. After 1 hour, the reaction mixture was removed and subjected to high vacuum to remove THF. The yellow solution was then chromatographed on

a reverse phase column (35 x 120 mm), eluted with 300 ml water followed by 100 ml portions of 5, 10, 15 .... 30% acetonitrile-water. Lyophilization of the desired fractions gave the p-nitro-benzyl ester as a yellow solid in an amount of 930 mg.

This was transferred to a pressure flask containing 25 ml of ether, 25 ml of THF and 25 ml of phosphate buffer, prepared by dissolving potassium dihydrogen phosphate (1.36 g, 0.01 mol) in 100 ml of water and adjusting the pH to 7.4 by addition of 45% aqueous KOH) and 900 mg of 10% palladium-on-charcoal.

The hydrogenation was carried out at 40 psi for 1 hour after which the organic phase was separated and washed with 2 x 5 ml of water. The combined aqueous phases were filtered and concentrated under high vacuum. The remaining solution was chromatographed on a reverse phase column (35 x 120 mm) eluted with water. Carbapenem-containing fractions were combined and lyophilized to give 1.21 g of a slightly greenish solid. This was purified by HPLC (10 x 300 mm water Microbondapack C-18 column, H 2 O as eluent) to give the pure title product, 480 mg (41%):<1>HNMR (D 2 O)

6: 1.23 (3H, d, J=6.4 Hz), 3.11 (2H, d, J=9 Hz), 3.37 (1H,

q, J=3.0, 6.1 Hz), 4.02 (7H, m), 5.18 (2H, s), 8.53 (1H, s): IR (nujol mull) 1750 cm"< 1>: UV (f osf atpuf fer, pH 7.4UITlcLKS (e = 7,810) .

Example_el_5

3-/ 5-( 1, 4- dimethyl- 1, 2, 4- triazolium) methanethio7- 6a-(/ T-( R)-hydroxyethyl7- 7- oxo- 1- azabicyclo/ 3. 2. 07hept- 2- en- 2- carboxylate

A. 1- methyl- 5- methanethiolacetate- 1, 2, 4- triazole

Methanesulfonyl chloride (0.46 mL, 6.0 mmol) was added dropwise to an ice-cooled, stirred solution of 1-methyl-5-hydroxymethyl-1,2,4-triazole (565 mg, 5.0 mmol) and triethylamine (0 .91 ml, 6.5 mmol) in 5 ml methylene chloride. After 30 minutes additional triethylamine (1.05 mL, 7.5 mmol) was added followed by thiolacetic acid (0.53 mL, 7.5 mmol) and stirring was continued for 45 minutes. The reaction mixture was diluted with methylene chloride and washed with water. The aqueous phase was extracted with 3 x 5 ml of methylene chloride and the combined organic phases were dried over MgSO 4 and the solvent removed. Column chromatography on silica gel gave pure 1-methyl-5-methanethiolacetate-1,2,4-triazole (570 mg) as a yellow oil (in addition, they were rechromatographed (preparative TLC, silica gel) an impure fraction of 200 mg to obtain additional 100 mg pure substance (overall yield: 85%)),<1>HNMR (CDCl3) 6: 2.38 (3H, s), 3.90 (3H, s), 4.25 (3H, s), 7.80 (1H, p).

B. 3-/ 5- Q , 4- dimethyl- 1 , 2 , 4- triazolium) methanethio7- 6a-(/ T - ( R ) -

hydroxyethyl 7- 7- oxo- 1- azabicyclo/ 3. 2. 07hept- 2- en- 2-carboxylate

Methyl trifluoromethanesulfonate (1.20 mL, 10.7 mmol) was added dropwise to an ice-cooled solution of 1-methyl-5-methanethiolacetate-1,2,4-triazole (730 mg, 4.27 mmol) in methylene chloride (7 mL) . The reaction mixture was allowed to warm slowly to room temperature over 3 hours after which it was concentrated. The remaining oil was triturated with ether to give an impure 1,4-dimethyl-5-methanethiol acetate-1,2,4-R.G. Jones and C. Ainsworth, J.Amer.Chem.Soc., 1955, 77, 1938. triazolium trifluoromethanesulfonate (1.46 g), which was used directly.

A solution of sodium hydroxide (512 mg, 12.8 mmol) in 5 mL of water was added to an ice-cooled solution of the triazolium salt (1.45 g, 4.35 mmol) in 5 mL of water. After 45 minutes, this solution was diluted to 25 ml with water and the pH adjusted to 7.6 with solid potassium dihydrogen phosphate. This solution was then added to an ice-cooled stirred solution of enol phosphate (2.00 g, 3.45 mmol) in 25 mL of THF. After 30 minutes, the reaction mixture was transferred to a pressure bottle containing 40 ml of ether and 2.0 g of 10% palladium-on-charcoal. This mixture was hydrogenated under 45 psi for 1 1/4 hours. The reaction mixture was then diluted with

25 ml of ether and filtered. The organic phase was separated and washed with 2 x 5 ml of water. The combined aqueous phases were washed with 3 x 25 mL of ether and then concentrated in vacuo. Column chromatography (reverse phase, 45 x 130 mm, water as eluent) followed by lyophilization of the carbapenem-containing fractions gave 650 mg of impure material. This was rechromatographed to give pure title product (450 mg, 39%): <1>HNMR (D 2 O ) 6 : 1.24 (3H, d, J=6.4 Hz), 3.19 (2H, q, J=2.6, 9.2 Hz), 3.45 (1H, q, J=2.8, 6.0 Hz), 3.91 (3H, s), 4.06

(3H, s), 4.08-4.36 (2H, m), 4.54 (2H, d, J=2.8 Hz), 8.71 (1H, s); IR (nujol mull) 1755 cm"<1>; UV (phosphate buffer, pH 7.4) X max 294 nm (e = 8'202)'* T£ (phosphate buffer, pH 7.4, M=0.067, T= 37°C) 9.1 h.

Example_el_6

( 1' R, 5R, 6S)- 3-/ 71, 3- dimethyl- 5- tetrazolium)- methylthio7- 6-( 1 - hydroxyethyl)- 7- oxo- 1- azabicyclo/ 3. 2. o7hept- 2- en- 2- carboxylate

A. 5- Carbethoxy- 2- methyltetrazole and 5- Carbethoxy- 1- methyltetrazole

1 a. Methylation of diazomethane

A solution of 5-carbethoxytetrazole 1 (9.17 g, 0.064 mmol) in 80 mL of ethyl ether<2> was cooled to 0°C and treated dropwise over 15 min with a solution of diazomethane (3 g, 0.071 mmol) in 200 mL ether. The pale yellow solution was stirred for 30 minutes and excess diazomethane destroyed by addition of acetic acid (1 mL). Evaporation of the solvent and distillation of the residue gave a clear oil with a boiling point of 95-100°C/0.5 torr; 9.64 g (96%).<1>Hmr showed a mixture of 1-methyl and 2-methyl isomers in a ratio of 6:4. Separation of the two isomers could not be carried out either by d1 distillation or by HPLC: v mak, s: 1740 cm (C=0 of ester): Hmr (CDCl 3 ) 6: 1.53 (3H, two overlapping t, J= 7.0, CH2CH3), 4.46 and 4.53 (3H, 2S, CH3 of 1-methyl- and 2-methyltetrazoles, ratio 6:4. Methylene of the 2-isomer is in a lower field and is the smallest product) , 4.5 ppm (2H, two overlapping q, CH^CH3.

1b. 5- carbethoxy- 2- methyltetrazole

D. Moderhack, Chem. Ber., 108, 887 (1975)<2>The use of a mixture of ethanol and ether gave the same ratio isomers. A mixture of 5-carbethoxy-2-methyltetrazole and 5-carbethoxy-1-methyltetrazole (0.252 g, 1.61 mmol, ratio of the two isomers 1:1) in 0.5 mL of iodomethane was sealed in a glass tube and op< p>heated to 100°C for 15 hours and at 130°C for 6 hours. Distillation of the reaction mixture gave the title compound as a pale yellow oil: 0.139 g (55%); boiling point 95-100°C/ -1

0.5 torr (air* ftbath temperature): ir (film)v IT13.K , S: 1740 cm (C=0 of ester); Hmr (CDClg) 6: 1.46 (3H, t, J=7.0, CH3CH2), 4.53 (3H, s, CH3-2), 4.5 (2H, q, J=7.0, CH2CH3).

2. Methylation with dimethyl sulfate

A solution of 5-carbethoxytetrazole (1.42 g, 0.01 mol) in 20 mL of dry acetone was treated with anhydrous potassium carbonate (1.38 g, 0.01 mol) and dimethyl sulfate (1.26 g, 0.01 mol ). The mixture was heated under reflux for 12 hours. The carbonate was filtered and the solvent evaporated under reduced pressure. The residue was diluted with 30 ml of dichloromethane, washed with 10 ml of saturated sodium bicarbonate, 10 ml of brine and dried over anhydrous sodium sulfate. Evaporation of the solvent and distillation under vacuum gave a clear oil: 1.46 g (93%); boiling point 85-110°C/0.5 torr. Hmr showed the presence of 2 isomers in a 1:1 ratio.

B. 5- hydroxymethyl- 2- methyltetrazole

1. When reducing the ester mixture

A mixture of 5-carbethoxy-1-methyltetrazole and 5-carbethoxy-2-methyltetrazole (ratio 6:4) (7.60 g, 0.049 mol)

in 50 mL of dry tetrahydrofuran was cooled to 0°C and treated with lithium borohydride (1.06 g, 0.049 mmol) added in small portions over 15 min. The mixture was kept at 10°C for 30 minutes of addition and then stirred at 20°C for 4 hours. The mixture was cooled to 0°C and the excess hydride was carefully

readily destroyed by the addition of 6N HCl (pH 7 after no more gas is evolved). The solvent was concentrated under vacuum and the remaining oil diluted with 200 ml of dichloromethane, washed with 10 ml of brine and finally dried over Na 2 SO 4 . Concentration of the solvent and distillation of the residue under vacuum gave 1.83 g corresponding to 33% of a clear oil.<1>Hmr of this substance showed that the product was 5-hydroxymethyl-2-methyltetrazole.

2. By reduction of 5-carbethoxy-2-methyltetrazole

To a solution of 5-carbethoxy-2-methyltetrazole (0.139 g, 0.89 mmol, obtained by isomerization of the ester mixture with methyl iodide) in 1 ml of dry tetrahydrofuran at 10°C was added lithium borohydride (0.019 g, 0.87 mmol ). The mixture was slowly warmed to room temperature and stirred for 4 hours. Excess borohydride was destroyed by careful addition of 6N HCl at 0°C (pH 7). The solvent was evaporated and the residue dissolved in 25 ml of dichloromethane

and dried over anhydrous sodium sulfate. Evaporation of the solvent gave the title compound as a clear oil: 0.092 g (91%); boiling point 90-120°C/0.5 torr with decomposition;

ir (film) vmax: 3350 cm"<1>(broad, OH);<1>Hmr (CDClj) 6: 4.4 (2H, s, CH3-2), 4.93 (2H, s, CH2" 5).

C. 5- acetylmercaptomethyl- 2- methyltetrazole

To a solution of 5-hydroxymethyl-2-methyltetrazole (1.83 g, 1.1.7 mmol) in 25 ml of dry dichloromethane at 0°C was

added methanesulfonyl chloride (1.47 g, 12.9 mmol) followed by triethylamine (1.30 g, 12.9 mmol) added dropwise over 5 minutes.

The mixture was stirred at 0°C for 1 h and then treated with a solution of potassium thioacetate (1.60 g, 14.0 mmol) in

10 ml of dry N,N-dimethylformamide. The resulting gel was stirred at 0°C for 3 hours. The reaction mixture was diluted with 200 ml of dichloromethane, washed with 20 ml of brine and dried over anhydrous sodium sulfate. Evaporation of the solvent under vacuum and chromatography of the resulting oil over silica gel (2x15 cm, eluting with dichloromethane and dichloromethane acetone 5%) gave the title compound as a clear oil: 1.31 g (65%); ir (film) vmax: 1696 cm-1 (C=0 of thioester);<1>Hmr (CDCl3) <5 : 2.43 (3H, s, SAc), 4.36 (3H, s, 2-CH3 ), 4.38 ppm (2H, s, 5-CH2). D. 5- mercaptomethyl- 1, 3- dimethyltetrazolium trifluoromethanesulfo- nat A solution of 5-acetylmercaptomethyl-2-methyltetrazole (0.400 g, 2.32 mmol) in 3 mL of dry dichloromethane was treated with methyl triflate (0.76 g, 4.64 mmol) and stirred at 22°C for 16 h. This salt was dissolved in 5 mL of cold, oxygen-free water and treated with 4 M sodium hydroxide (0.8 mL, 3.2 mmol). The mixture was stirred at 0°C for 40 minutes, diluted with 7 ml of water after which the pH was adjusted to 7.3 with saturated Kf 2 PO 3 . The resulting clear solution was kept under nitrogen and used immediately in the subsequent step.

E. ( 1' R, 5R, 6S)- 3/ T, 3- dimethyl- 5- tetrazolium)- methylthio7- 6-( 1- hydroxyethyl)- 7- oxo- 1- azabicyclo/ 3. 2. 07hept- 2 - one-2-carboxylate

A solution of enol phosphate (0.915 g, 1.58 mmol) in

8 mL of tetrahydrofuran was cooled to 0°C and treated dropwise with a solution of 5-mercaptomethyl-1,3-dimethyltetrazolium trifluoromethanesulfonate (2.32 mmol, prepared above)

within 20 minutes. The pH value of the reaction was stable at 6.5 throughout the addition. After 20 additional minutes, the pH of the solution was adjusted to 7.0 with saturated sodium bicarbonate. The mixture was transferred to a hydrogenation flask diluted with 10 ml of THF, 20 ml of ether and 20 g of ice. The carbapenem was hydrogenated over 10% palladium-on-charcoal under 45 psi while the temperature was slowly raised to 22°C for 90

minutes. The catalyst was separated from and washed with 5 ml

cold water and 20 ml of ether. The aqueous phase was washed with 20 ml of ether and kept under vacuum for 20 minutes to remove traces of organic solvent. Chromatography on PrePak 500-C/ 18 eluting with water gave the title compound as a white

powder after lyophilization: 0.266 g (49%); /α7^<3>+ 13° (c 1.04, H 2 O); UV (H20, pH 7.4) *max: 294 nm (e7,500); ir (KBr) Vmax: 1755 (C=0 of B-lactam), 1600 cm"1 (broad, C=0 of carboxylate);<1>Hmr (D20) 6: 1.24 (3H, d, J= 6.4Hz, CH3CHOH), 3.0-3.3 (2H, m, H-4), 3.42 (1H, dd, J=5.8, J=2.9, H-6), 4 -4.2

(2H, m, H-5 and CH3CHOH), 4.34 and 4.57 (2x 3H, 2S, CHg-1 and 3 of tetrazole), 4.49 and 4.51 (2H, 2s, CH-S) . The product has a

o -4 half-life of 10.5 hours at 37 C (c of 10 M in pH 7.4 phosphate buffer.

Example_el_7

Preparation of 3-(N-methylpyridin-2-yl-methanethio)-6a-/1-(R)-hydroxyethyl7-7-oxo-1-azabicyclo/3.2.07hept-2-ene-2-carboxylate

Via a "one-flask" process

A. Preparation of enol phosphate ( 2) An ice-cooled solution of ketone 1 (3 g, 8.62 mmol) in 30 ml of acetonitrile is treated with (9 mmol, 1.04 equivalents, 1.57 ml) ethyldiisopropylamine (addition time approx. 2 minutes ) and chlorodiphenyl phosphate (9 mmol, 1.04 equivalents, 1.87 ml) (addition time approx. 2 minutes). The reaction mixture was stirred for 45 minutes and TLC (ethyl acetate, silica gel) showed that ketone 1 disappeared. The solution was diluted with 60 ml of ethyl acetate, washed with 2 x 50 ml of cold water and brine, dried over sodium sulfate and concentrated (bath temperature below 20°C) to form a foam which was used as such. B. Preparation of thiol ( 4)

An ice-cooled solution of thioacetate 3 (3.31 g, 10 mmol) in water purged with nitrogen for 5 min was treated dropwise (about 5 min) with a cooled solution of sodium hydroxide (1.75 equivalents, 17.5 mmol, 0 .7 g) in 8 ml of water. The mixture turned yellow. After 75 minutes under nitrogen, the pH value was adjusted to 7.4 with saturated aqueous solution of Kf^PO^. The reaction mixture was diluted with 15 ml of water. This aqueous solution of thiol 4 (50 mL, 0.2 mmol/mL) was used as such.

C. Coupling

An ice-cooled solution of compound 2 (impure, prepared under A, 8.62 mmol) in 50 ml tetrahydrofuran was treated dropwise with the aqueous solution of thiol 4 prepared in B (5 ml solution every 5 minutes). During the course of the reaction, the pH value of the reaction mixture was kept at approx. 6.5-7.5 (preferably 7) by adding cooled 2N sodium hydroxide solution. The reaction mixture was followed by TLC (a) silica gel, ethyl acetate; (b) reverse phase Analtech RPSF, CHgCN-pH 7 buffer (4:6).

Finally, 1.15 equivalents of thiol (50 ml solution) were used. The reaction was complete after 1 hour at 0°C and the mixture was used as such for hydrogenation after the pH value was adjusted to 7.

The reaction mixture containing 5 (prepared under C) was transferred to a Parr flask with 10 mL of THF, phosphate buffer (pH 7, 0.1 M) (10 mL), 75 mL of ether, and 5 g of Pd-C 10% and hydrogenated at 45 psi at 3-10°C for 2 hours. The catalyst was filtered off, the whole washed with 3 x 10 ml of water and the pH carefully adjusted to 6.2 with cold 2N NaOH. Ether was added and the aqueous phase separated from and washed again with ether. The aqueous phase was stripped of organic solvent under vacuum and then purified on a Bondapak C-18 column (100 g, 4.5 x 13 cm) with cold distilled water. The pale yellow fractions containing the product (checked by U.V. and TLC) were lyophilized to give 1.46 g or 50%<*>of 6 as a yellow powder. X269, e = 9000, X271 , e=11064.

<*>Yield calculated from bicyclic ketone.

Example_el_8

If the keto intermediate 1 in the method in example 7 is replaced with an equimolar amount of the corresponding 1B-methyl intermediate, the carbapenem end product shown above is obtained.

Example_el_9

If the keto intermediate 1 in the method in example 7 is replaced by an equimolar amount of the corresponding 1a-methyl intermediate, the carbapenem end product shown above is obtained.

Claims (2)

1. Procedure for preparing a compound of the formula 8 1 wherein R is hydrogen and R is selected from the group consisting of hydrogen; substituted and unsubstituted alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl and cycloalkylalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the heteroatom or atoms in the above-mentioned heterocyclic part are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl parts which are connected to the heterocyclic parts have 1-6 carbon atoms; wherein the substituent or substituents with respect to the above-mentioned groups are independently selected from the group consisting of C 1 -g alkyl which is optionally substituted with amino, halogen, hydroxy or carboxyl halogen wherein with respect to the above-mentioned substituents, the groups R 3 and R 4 are independently selected from hydrogen; alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl parts; phenyl; aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl parts in connection with these hetero-3 4 cyclic parts have 1- 6 carbon atoms or R and R form, together with the nitrogen atom to which at least one is attached, a 5- or 6-membered, nitrogen-containing, heterocyclic ring, R 9 is as defined for R 3 except that it cannot be hydrogen; or wherein R 1 and R 8 together form the C 1 -C 4 alkylidene substituted with hydroxy, R 1 is selected from the group consisting of substituted and unsubstituted alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl and cycloalkylalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the heteroatom or atoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms, and the alkyl moieties connected to the heterocyclic moieties have 1-6 carbon atoms; in which the above-mentioned R 5 groups are optionally substituted with 1-3 substituents independently selected from: Cj -Cg alkyl which is optionally substituted with amino, fluorine, chlorine, carboxyl, hydroxy or carbamoyl; fluorine, chlorine or bromine: phenyl, which is optionally substituted with 1-3 fluorine, chlorine, bromine, C..-C -alkyl, -OR3, -NR3R4, -S0oR3, -CO~R3 or -CONR3R4, 6 3 4 9 5 3 2 in which R , R and R in these R -substituents are as defined 5 above; or R may be bound to at another point on the ring, so that a fused heterocyclic or heteroaromatic ring is formed which may contain additional heteroatoms selected from 0, S and N;R^ is selected from the group consisting of hydrogen, substituted and unsubstituted alkyl, alkenyl and alkynyl with 1 -10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl1 with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl parts, spirocycloalkyl with 3-6 carbon atoms, phenyl, aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has 1-6 carbon atoms, heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1-4 oxygen, nitrogen and sulfur atoms and the alkyl moieties, which are connected to the heterocyclic moieties, have 1 -6 carbon atoms, in which the substituent or substituents with respect to the above-mentioned groups are selected from the group consisting of amino, mono-, di- and trialkyla mino, hydroxyl, carboxyl, mercapto, alkylthio, phenylthio, sulfamoyl, amidino, guanidino, nitrogen, chlorine, bromine, fluorine, cyano and carboxy, and in which the alkyl parts of the above-mentioned substituents have 1-6 carbon atoms, A is C. I —b, straight chain or branched alkylene, R 2 is hydrogen, an anionic charge or a conventional easily removable carboxyl protecting group, provided that when R 2 is hydrogen or a protecting group, a counterion is also present ; and means a substituted or unsubstituted mono-, bi- or polycyclic, aromatic, heterocyclic group containing at least one nitrogen atom in the ring, which ring is bonded to A through a ring carbon atom and has a ring nitrogen quaternized by the group R~*, or a pharmaceutically acceptable salt thereof, characterized in that an intermediate product with the formula is reacted 18 15 2 1 wherein R , R , and R have the meanings given above, R is a conventional, easily cleavable carboxyl protecting group and L is a conventional cleavable group, with a thiol compound of the formula in which A and have the meanings given above and X © is an opposite anion, in an inert solvent and in the presence of a base to form a carbapenem product of the formula 1 8 1 5 2 1 in which R, R, R, R ,A 0 and X have the meanings given above, and if desired, the carboxyl protecting group R 2 1 is removed to form the corresponding deblocked compound of formula (I) or a pharmaceutically acceptable salt thereof. 2. Process according to claim 1, characterized in that the base is sodium hydroxide. 3. Method according to claim 1, characterized in that the reaction is carried out at a temperature in the range between -15°C and +15°C. 4. Method according to claim 1, 2 or 3, characterized in that the cleavable group L is chlorine, bromine, iodine, benzenesulfonyloxy, p-toluenesulfonyloxy, p-nitrobenzenesulfonyloxy, methanesulfonyloxy, trifluoromethanesulfonyloxy, diphenoxyphosphinyloxy or di(trichloroethoxy)phosphinyloxy . 5. Method according to claim 1, 2 or 3, characterized in that the cleavable group L is diphenoxyphosphinyloxy. 6. Method for producing a compound with the formula wherein R 2 is hydrogen, an anionic charge or a conventional easily removable carboxyl protecting group, provided that when R 2 is hydrogen or a protecting group, a counter ion is also present, or a pharmaceutically acceptable one thereof, characterized by that one converts an intermediate product with the formula where L is a conventional leaving group and R 2 'is a conventional easily removable carboxyl protecting group, with a thiol compound with the formula where X is an opposite anion, in an inert solvent and in the presence of a base to prepare a carbapenem product of the formula 21 (°) where R and X 2 have the meaning given above, and if desired, the protecting group R 2 ′ is removed to form the corresponding deblocked compound of formula (I) or a pharmaceutically acceptable salt thereof. 7. Procedure for preparing a compound of the formula where R 2 is hydrogen, an anionic charge or a conventional, easily removable carboxyl protecting group, provided that, when R 2 is hydrogen or a protecting group, an opposite is also present, or a pharmaceutically acceptable salt thereof, characterized by that one converts an intermediate product with the formula where L is a conventional leaving group and R 2 1 is a conventional easily removable carboxyl protecting group, with a thiol compound of the formula where X® is an opposite anion, in an inert solvent and in the presence of a base for the preparation of a carbapenem- product with the formula 2 • X ( <=> ) meaning, where R and X w have the meaning given above, and if desired, the protecting group R 2 ' is removed to form the corresponding deblocked compound of formula (I), or a pharmaceutically acceptable salt thereof .