CH658248A5 - METHOD FOR PRODUCING CARBAPENEM DERIVATIVES. - Google Patents

Description

The invention relates to processes for the preparation of carbapenem derivatives and to quaternary aminothiol compounds as agents for carrying out this process. The carbapenem derivatives produced according to the invention have the formula

0 0 NR-1

-OR3, -OCNR3R4, -CNR3R4, -NR3R4, -0,

NR3R4

0 0

20 -s02nrjrh, -nhcnr ^ r, r cnr -, -CO ^ ß? , = 0, 0 0 0

-ocr3, -sr3, -sr9, -sr9, -cn, -nj, -oso ^ r3,

0

25-0S02R3, -NR3S02R4, -NR3C = NR4, -NR3C02R4 or r3

, 8 h

S-A-R '

COOR

14

(I)

wherein A represents a cyclopentylene, cyclohexylene or C ^ alkylene radical, which is optionally substituted by one or more CM alkyl groups, R14 represents an aromatic or non-aromatic heterocycle containing a quaternized nitrogen atom and via a quaternary Nitrogen atom is bonded to A, R2 represents a hydrogen atom, an anionic charge or a carboxyl protective group, where, if R2 denotes hydrogen or a protective group, a counter anion is also present, and R1 and R8 have the meaning given in claim 1.

The carpabenem derivatives produced by the process according to the invention are described in US patent applications Serial Nos. 366,910,471379 and 389,652 (inventors: Choung U. Kim and Peter F. Misco Jr.). The entire disclosure of each of these applications is hereby incorporated by reference.

US Application No. 366,910 and its continuation application, Serial No. 471379, which corresponds to DE-OS 3 312 533, describes the preparation of carbapenem antibiotics of the formula

(IA)

35

COOR

wherein

R8 represents a hydrogen atom and R1 represents a hydrogen atom or represents the following substituted and unsubstituted radicals:

an alkyl, alkenyl and alkynyl group having 1 to 10 carbon atoms; a cycloalkyl and cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl units; a phenyl group; an aralkyl, aralkenyl and aralkynyl group, in which the aryl

-no2,

wherein in the above-mentioned substituents, the groups 30 R3 and R4 independently of one another for a hydrogen atom; an alkyl, alkenyl and alkynyl group having 1 to 10 carbon atoms; a cycloalkyl, cycloalkylalkyl and alkylcycloalkyl group having 3 to 6 carbon atoms in the cycloalkyl ring and having 1 to 6 carbon atoms in the alkyl units; a phenyl group; an aralkyl, aralkenyl and aralkynyl group, wherein the aryl unit is a phenyl group and the aliphatic part has 1 to 6 carbon atoms; or a heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl group, where the heteroatom or the heteroatoms in the above-

40 named heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl units connected to said heterocyclic units have 1 to 6 carbon atoms, or R3 and R4 together with the nitrogen atom to which at least one is attached

45 of these radicals are bonded to form a 5- or 6-membered nitrogen-containing heterocyclic ring;

R9 is defined as the group R3, but cannot denote a hydrogen atom;

or in what

50 R1 and R8 together mean a C2_io-alkylidene radical or a C2_io-alkylidene radical substituted by a hydroxyl group;

A represents a cyclopentylene, cyclohexylene or a C 1-4 alkyl group which is optionally substituted by one or more C 1 -C 4 alkyl groups;

55 R2 represents a hydrogen atom, an anionic charge or a conventional, easily removable carboxyl protective group, with the proviso that if R2 represents a hydrogen atom or a protective group, a counter anion is also present, and the rest of the formula

60

-N

represents a substituted or unsubstituted, mono-, bi- or poly-65 cyclic, aromatic, heterocyclic radical which contains at least one nitrogen atom in the ring and is bonded to A via a ring nitrogen atom, thereby forming a quaternary ammonium group;

658 248

4th

and their pharmaceutically acceptable acid addition salts. wherein R8, R1, R3, R4 and R '; the meanings given above These compounds are prepared by a process, have:

which is shown in the following reaction scheme: Rs is selected from the following substituted and unsubstituted

groups: alkyl, alkenyl and alkynyl with 1 to 10 carbon-5 atoms; Cyclkoalkyl and Cycloalkylalkyl with 3 to 6 carbon diphenylchlorophosphate substance atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl units; Phenyl; Aralkyl, aralkenyl and aralkinyl,

R8 H

• COOR

III

(° C6H5) 2

HS-A-OH

->

COOR

wherein the aryl moiety is phenyl and the aliphatic portion has 1 to 6 carbon atoms; Heteroaryl, heteroaralkyl, 10 heterocyclyl and heterocyclylalkyl, wherein the heteroatoms or the heteroatom in the abovementioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl units connected to the heterocyclic units mentioned have 1 to 6 carbon atoms 15;

the above-mentioned R5 residues if desired by

R8 H

VI

A = alkylene or c -c-cycloalkylene ibis 3 substituents which are selected independently of one another - 5 5 from:

a C 1-4 alkyl radical which is optionally substituted by amino, fluorine, chlorine, carboxyl, hydroxy or carbamoyl -A-OH methanesulfonyl chloride;

Fluorine, chlorine or bromine;

-OR3; -OCO, R3; -OCOR3; -OCONR3R4; -OSO, R3; -Oxo; -NR3R4; R3CONR4; -NR3CO, R4; -NR3CONR3R4; -NR3S02R4; 25 -SR3;

O

Î

-S-R9;

> 30 ^ »° g

-S-R

-SO3R3; -C02R3; -CONR3R4; -CN; or a phenyl radical which is optionally substituted by: 1-3-fluorine, chlorine, bromine, C 1-4 alkyl, -OR3, -NR3R4, -S03R3, 35 -C02R3 or -CONR3R4, where R3, R4 and R9 in these substituents R5 have the meanings given above;

or wherein R5 is a divalent phenylene or Cw alkylene

group that means with the ring:

^ • COOR2

S-A-0S02CH3

COOR

O

II

COOR

(jP = counter anion) 40 ®

is connected to form a bridged polycyclic group;

A represents a cyclopentylene or cyclohexylene radical or a C2 ^ -. 43 alkylene group which is optionally substituted by one or more CM alkyl groups;

R2 represents a hydrogen atom, an ionic charge or a conventional, easily removable carboxyl protective group, with the proviso that if R2 represents a hydrogen atom or a 50 protective group, a counterion is also present; and the rest of the formula

%O

55 denotes a substituted or unsubstituted, mono-, bi- or poly-cyclic, non-aromatic, heterocyclic radical which contains at least one nitrogen atom in the ring and is bonded to A via a ring nitrogen atom in US Pat. No. 389,652 and the continuation application, whereby a quaternary 499 690 describe the production of carbapenem antibioammonium group;

tika of the general formula IB 60 and the pharmaceutically acceptable salts thereof. This l'A

; 8 h

S

-A-I

Process is shown in the following reaction scheme:, 8 H

COOR2

Diphenyl chlorophosphate

COOR '

2 '

IB

III

5

658 248

o II

P (° C6H5) 2

0

, 8 h

N - IV

COOR "

HS-h-OH

In addition, the quaternization is carried out with the entire carbape-nem compound.

The object of the present invention is therefore to prepare a new process for preparing the compounds IA or IB.

S-A-OH

'COOR *

A = AlJcylen- or c5 ~ Cg cycloalkylene 5, which (1) has fewer reaction stages, (2) leads to higher yields, (3) first enables the formation of the quaternized amine, which is then later synthesized on the carbapenem Core is bound, and (4) easier formation of quaternary amine products with a variety of io amines, e.g. B. sterically hindered amines or those with methanesulfonyl chloride low pKb values.

9 The invention relates to those in claims 1 and 2

defined new processes for the preparation of the compounds of the formula I 'or I ". These compounds can be described by the formula I below:

• S-A-0S02CH3

COOR

î8 H

20th

T I

S-A-R 2

COOR "

14

VI

R H

II

IB

S-A-I

COOR

COOR

ÎO,

Ag + X6 ^

possibly De-. blockage ^

wherein

25 R8 represents a hydrogen atom and

R1 represents a hydrogen atom or represents the following substituted and unsubstituted radicals:

an alkyl, alkenyl and alkynyl group having 1 to 10 carbon atoms; a cycloalkyl and cycloalkylalkyl group having 30 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl units; a phenyl group; an aralkyl, aralkenyl and aralkynyl group, wherein the aryl unit represents a phenyl group and the aliphatic part has 1 to 6 carbon atoms; a heteroaryl, heteroaralkyl, 35 heterocyclyl and heterocyclylalkyl group, the hetero atom or the heteroatoms in the above-mentioned heterocyclic units being selected from 1 to 4 oxygen, nitrogen and sulfur atoms and the alkyl units connected to the heterocyclic units 1 to 6 carbon atoms have 40;

where the substituent or the substituents of the abovementioned radicals mean one of the following groups:

a C 1-4 alkyl group which is optionally substituted by amino, halogen, hydroxy or carboxyl, 43 a halogen atom,

In this known process, the compound of the formula III is used and reacted in an inert organic solvent with diphenylchlorophosphate in the presence of a base to give the intermediate IV. The intermediate compound IV is then reacted with a mercaptan reagent of the formula HS-A-OH in an inert, organic solvent and in the presence of a base to give the intermediate compound V. Intermediate V is then aylated with methanesulfonyl chloride in an inert organic solvent and in the presence of a base to give intermediate VI, which is reacted with an iodide ion source in an inert organic solvent to form intermediate II. This intermediate compound II is reacted with the desired amine in an inert organic solvent and in the presence of silver ions to give the quaternized product I'A or I'B, which can then be deblocked, giving the corresponding deblocked carbapenem compound of the formula IA or IB receives.

The method described above has several disadvantages. So this process runs over different stages, and it would be advantageous if the number of stages could be reduced. The overall reaction yield is also quite low.

NO

50

55

-or3, -ocnr3r / f, -cnr3r4, -nr3r4t -ù,

vnrV

0 0

U n t, L L 'S

-s02nr:> r, -nhcnr ^ r, r-'cnr -, -cogr, = 0,

0 0 0

ti * anqiiq

-ocr, -sr, -sr, -sr, -cn, -n ,, -oso ^ r-1,

n y y

O

-0s02r3, -nr3s02r4, -nr3c = nr4, -nr3c02r4 or r3

-no2,

fio where in the above-mentioned substituents the groups

R3 and R4 independently represent a hydrogen atom; an alkyl, alkenyl and alkynyl group having 1 to 10 carbon atoms; a cycloalkyl, cycloalkylalkyl and alkylcycloalkyl group having 3 to 6 carbon atoms in the cycloalkyl ring having 65 1 to 6 carbon atoms in the alkyl units; a phenyl group; an aralkyl, aralkenyl and aralkynyl group, wherein the aryl unit is a phenyl group and the aliphatic part has 1 to 6 carbon atoms; or a heteroaryl, hetero

658 248

roaralkyl, heterocyclyl and heterocyclylalkyl groups, the heteroatom or the heteroatoms in the abovementioned heterocyclic units being selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl units connected to the said heterocyclic units having 1 to 6 carbon atoms, or are

R3 and R4 together with the nitrogen atom to which at least one of these radicals is bonded form a 5- or membered nitrogen-containing heterocyclic ring;

R9 is defined as the group R3, but cannot denote a hydrogen atom;

or in what

R1 and Rs together represent a C2_i0-alkylidene or Q.jo-alkylidene radical which is substituted by a hydroxyl group;

A represents a cyclopentylene, cyclohexylene or a C2_6-alkylene group which is optionally substituted by one or more C 1-4 alkyl groups;

R2 represents a hydrogen atom, an anionic charge or a carboxyl protecting group, with the proviso that if R2 represents a hydrogen atom or a protecting group, a counter anion is also present, and

R14 denotes a quaternized, nitrogen-containing, aromatic or non-aromatic heterocycle which is bonded to A via a ring nitrogen atom, whereby a quaternary ammonium group is formed,

or a pharmaceutically acceptable salt thereof.

The compounds of formula IV serve as starting materials

5;

10th

15 i,

20th

25th

30th

COOR

2 *

IV

wherein R1 and R8 have the meanings given above, R2 'represents a customary, easily removable carboxyl protective group and L represents a leaving group,

and the thiol compound of formula VII

HS-A-R14 X®

VII

wherein A and R14 have the meanings given above and X® represents a counter anion.

The invention also relates to thiol compounds of the formula VII as agents for the preparation of the carbapenem compounds of the formula I 'and I ".

These carbapenem compounds are effective antibacterial or intermediate compounds which are useful in the preparation of such agents.

The compounds of the general formula I contain the carpapenem core I

6

35

40

45

50

55

and can thus be referred to as l-carba-2-penem-3-carboxylic acid derivatives. However, the connections can alternatively be considered to have the following basic structure

4th

60

-

Have O x and are called 7-Oxo-l-azabicyclo [3.2.0] hept-2-en-2-carboxylic acid derivatives. According to the invention

65

Compounds comprises, in which the hydrogen atoms in the 5- and 6-position can be both in the ice and in the trans position, the preferred compounds having 5R, 6S (trans) configuration, as is also the case with thienamycin.

The compounds of the general formula I can be unsubstituted in the 6-position or substituted by the substituent groups described above for the other carbapenem derivatives. In particular, R8 represents a hydrogen atom and R1 can represent a hydrogen atom or a substituent that differs from hydrogen and is disclosed, for example, in European patent application 38 869 (see the definition of R6). Alternatively, R8 and R1 together can mean a Q ^ o-alkylidene radical or a substituted (z. B. by hydroxy) C2_io-alkylidene radical.

The definitions for R1 and R8 are described in more detail below:

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

(b) “Heteroaryl” denotes mono-, bi- and polycyclic, aromatic, heterocyclic groups which contain 1 to 4 O, N or S atoms. Preferred are 5- or 6-membered heterocyclic rings, such as thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, tetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazyl-pyrazyl, pyrazyl-pyrazole

(c) “Heterocyclyl” means mono-, bi- and polycyclic, saturated or unsaturated, non-aromatic, heterocyclic groups which contain 1 to 4 O, N or S atoms. Preferred are 5- or o-membered, heterocyclic rings such as morpholinyl, piperazinyl, piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrrolinyl, pyrrolidinyl, etc.

(d) “Halogen” means chlorine, bromine, fluorine and iodine atoms and preferably chlorine or bromine atoms.

The term "carboxyl protecting group" refers to a known ester group which has been used to block a carboxyl group during the chemical reactions described below and which, if desired, can be removed by methods which do not destroy the rest of the molecule. Such processes include, for example, chemical or enzymatic hydrolysis, treatment under mild conditions with chemical reducing agents, irradiation with UV light and catalytic hydrogenation. Examples of such ester-forming protective groups are benzhydryl, p-nitrobenzyl, 2-naphthylmethyl, allyl, benzyl, trichlorethyl, silyl, e.g. Trimethylsilyl, phenacyl, p-methoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl and C, _16-alkyl, e.g. B. methyl, ethyl or butyl. Protecting groups which can be hydrolyzed under physiological conditions are also suitable. These include, for example, pivaloyloxymethyl, acetoxymethyl, phthalidyl, idanyl and methoxymethyl. Particularly advantageous carboxyl protective groups are the p-nitrobenzyl group, which can be easily removed by catalytic hydrogenolysis, and the allyl group, which can be removed by Pd (P03) 4-catalyzed reaction.

The pharmaceutically acceptable salts include the non-toxic acid addition salts, e.g. B. salts with mineral acid, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid, etc., and salts with organic acids, such as maleic, vinegar, lemon, amber, benzoic, wine, fumaric, almond, Ascorbic, milk,

658 248

Gluconic and Malic Acid. Compounds of the general formula

I in the form of acid addition salts can be described as

, 8 h

a cyclohexylene residue cT ~

-A-R

'COOR

14

\

J

or a Q ^ -alkylene radical which, if desired, is substituted by R2 = H or a protective group or one or more C ^ -alkyl substituents.

Preferred substituents A are cyclopentylene, cyclohexylene or alkylene of the formula in which X® denotes the acid anion. The counter anion X® can be selected so that pharmaceutically acceptable salts are obtained for therapeutic purposes. In the case of the intermediate compounds of the formula I, however, X® can also be a toxic anion. In such a case, the ion can subsequently be removed or replaced with a pharmaceutically acceptable anion to obtain the final active product for therapeutic use. If acidic or basic groups are present in group R1 or on the quaternized R14 radical, then suitable base or acid salts of these functional groups are also included according to the invention, e.g. B. acid 'addition salts in a basic group and metal salts (e.g. sodium, potassium, calcium and aluminum), the ammonium salt and salts with non-toxic amines (e.g. trialkylamines, procain, dibenzylamine, 1-ephenamine, N-benzyl-ß-phenethyl- amine, N, N'-dibenzylethylenediamine, etc.) in an acidic group.

Compounds of the general formula I, in which R 2 represents a hydrogen atom, an anionic charge or a physiologically hydrolyzable ester group, and the pharmaceutically acceptable salts thereof, are useful antibacterial agents. The other compounds of the general formula I are valuable intermediates which can be found in the abovementioned, biologically active compounds can be transferred.

Preferred compounds of the general formula I are those in which R8 is a hydrogen atom and R1 is a hydrogen atom, CH3CH2-,

r i

-c-

10th

15

R

11

r12

I.

-c-

R13

wherein R10, R11, R12 and R13 each independently represent a hydrogen atom or a C 1-4 alkyl group. A preferred embodiment comprises those compounds of the general formula I in which the substituent A is -CH2CH2-,

-chch2-

ch-r

-P

25th

or

-ch, ch-ch-z

CH

CH, OH v3 i

. CH-

'C-

or

OH

I.

CH3CH-

CH.

stands.

CH.

stands.

If the substituent A in some compounds of the general formula I is a cycloalkylene or branched alkylene radical, one or more additional, asymmetric carbon atoms are present, which leads to the formation of diastereoisomers. According to the invention, the mixtures of these diastereoisomers and also the individual, purified diastereoisomers are discussed.

The quaternized substituent R14 is an optionally substituted, mono-, bi- or polycyclic, aromatic or non-aromatic, heterocyclic radical which contains at least one nitrogen atom in the ring and is bonded to A via a ring nitrogen atom 40, whereby a quaternary ammonium group is formed.

A preferred class of substituents R14 are those of the general formula

In this subclass, the preferred compounds are those in which R1 is

45

oh yeah

-o stands. Particularly preferred connections have the absolute configuration 5R, 6S, 8R.

Also preferred are those compounds of the general formula I in which R1 and R8 together represent an alkylidene radical of the general formula hoch0

CH,

c =

This formula represents a substituted or unsubstituted, mono-, bi- or polycyclic heteroaryl radical which contains at least one nitrogen atom in the ring and is bonded via a ring 50 nitrogen atom to a carbon atom of the substituent A, resulting in a quaternary ammonium group. The heteroaryl radical can, if desired, be substituted by the following substituents: Q ^ alkyl, Q ^ alkyl substituted by hydroxy, amino, carboxy or halogen, 55 C ^ cycloalkyl; Ci_4-alkoxy; C ^ alkylthio; Amino; Cw-alkylamino; Di (Cw alkyl) amino; Halogen; CM alkanoylamino; C ^ alkanoyloxy; Carboxy;

0

II

form.

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

X /

H-C HC

2 \ /

H2C CH2

60

-C-0C1 _ ^ - alkyl;

Hydroxy; Amidino; Guanidino; Trifluoromethyl; Phenyl; Phenyl substituted by one, two or three amino, halogen, hydroxy, trifluoromethyl, ClJt alkyl or Q ^ alkoxy groups; Heteroaryl and heteroaralkyl, where the heteroatom or f> 5 the heteroatoms in the abovementioned heterocyclic units are selected from 1 to 4 O, N or S atoms and the alkyl group connected to the heteroaralkyl group has 1 to 6 carbon atoms.

658 248

8th

The heteroaryl radical bonded to the substituent A is preferably a 5- or 6-membered aromatic, heterocyclic radical which contains a quaternized nitrogen atom which is bonded directly to a carbon atom of the alkylene or cycloalkylene radical and, if desired, one or more additional heteroatoms which are selected are from O, N or S. Although in general any heteroaryl radical which is bonded to A via a quaternized nitrogen atom leads to biologically active carbapenem derivatives, a preferred embodiment of the invention comprises compounds in which the radical of the general formula

represents one of the radicals listed below:

y

(a) c

n or men are selected and the alkyl unit connected to said heteroaralkyl unit has 1 to 6 carbon atoms, or, if desired, are substituted so that a fused carbocyclic or heterocyclic ring is formed;

«1 I

(c)

10th

15

'-S

VT '

or

Q

wherein R5, R6 and R7 independently represent hydrogen; C1JS.-alkyl; C 1-4 alkyl substituted by hydroxy, amino, carboxy or halogen; C ^ cycloalkyl; Cw alkoxy; C 1-4 alkyl thio; Amino; C ^ -Alky lamino; Di- (C1-alkyl) amino; Halogen; CM-alkanoylamino; Çw-alkanoyloxy; Caboxy;

0

-C-0C1 ^ alkyl;

Hydroxy; Amidino; Guanidino; Trifluoromethyl; Phenyl; Phenyl substituted by one, two or three amino, halogen, hydroxy, trifluoromethyl, Cw-alkyl or CM-alkoxy groups; and heteroaryl and heteroaralkyl, wherein the heteroatom or the heteroatoms in the abovementioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl unit connected to said heteroaralkyl unit has 1 to 6 carbon atoms; or wherein two of the radicals R5, R6 or R7 together form a fused saturated carbocyclic ring, a fused aromatic carbocyclic ring, a fused saturated heterocyclic ring or a fused heteroaromatic ring;

(b)

if desired, these radicals are substituted on a carbon atom by one or more substituents, 20 which are independently selected from the following radicals: CM-alkyl; CM-alkyl substituted by hydroxy, amino, carboxy or halogen; C ^ cycloalkyl; CH alkoxy; CM alkylthio; Amino; C ^ -Alky lamino; Di- (C 1-4 alkyl) amino; Halogen; Cw Alkanoylamino; Cw alkanoyloxy; 25 carboxy;

0

-C-0C1_4-alkyl;

Hydroxy, amidino; Guanidino; Trifluoromethyl; Phenyl; Phenyl, 30 substituted by one, two or three amino, halogen, hydroxy, trifluoromethyl, Cw-alkyl or Cw-alkoxy groups; and heteroaryl or heteroaralkyl, where the heteroatom or the heteroatoms in the abovementioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl unit connected to said heteroaralkyl unit has 1 to 6 carbon atoms, or are so substituted if desired that a condensed carbocyclic or heterocyclic ring is formed;

35

40

(d)

ej.

Ì

II

N

45

50

© I

N ^ ^ 3

N. N

or

J

where the radicals are optionally substituted on a carbon atom by one or more substituents which are independently selected from the following radicals: C 1-4 alkyl; C 1-4 alkyl substituted by hydroxy, amino, carboxy or halogen; C ^ cycloalkyl; C, _} - alkoxy; Cw alkyl thio; Amino; C 1-4 alkylamino; Di- (C1_4-alkyl) amino; Halogen; C14 alkanoyloxy: C ^ alkanoylamino; Carboxy;

0

-C-OC1 _ ^ - alkyl;

Hydroxy; Amidino; Guanidino; Trifluoromethyl; Phenyl; Phenyl substituted by one, two or three amino, halogen, hydroxy, trifluoromethyl, C 1-4 alkyl or CM alkoxy groups; and heteroaryl or heteroaralkyl, where the heteroatom or heteroatoms in the heterocyclic units mentioned above contain from 1 to 4 oxygen, nitrogen or sulfur atoms.

if desired, these radicals are substituted on a carbon atom by one or more substituents which are independently selected from the following radicals 55: Cw-alkyl; C 1-4 alkyl substituted by hydroxy, amino, carboxy or halogen; Q ^ cycloalkyl; Cw alkoxy; C 1-4 alkyl thio; Amino; C 1-4 alkylamino; Di (Cw alkyl) amino, halogen, CM alkanoylamino; C ^ alkanoyloxy, carboxy;

0

60 II

-C-OC ^ _ ^ - alkyl;

Hydroxy; Amidino; Guanidino; Trifluoromethyl; Phenyl; Phenyl substituted with one, two or three amino, halogen, hydroxy, trifluoromethyl, CM-alkyl or C 1-4 alkoxy group 65 pens; and heteroaryl or heteroaralkyl, where the heteroatom or the heteroatoms in the above-mentioned heterocyclic units are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and those with the said heteroaralkyl-

9

658 248

unit-linked alkyl unit has 1 to 6 carbon atoms or, if desired, are substituted so that a fused carbocyclic or heterocyclic ring is formed;

N

\

»J-R

rv

(e)

or n-r e

// \

or r

where X is O, S or NR, where R C is alkyl or phenyl, these radicals being optionally substituted on a carbon atom by one or more substituents which are independently selected from the following radicals: CM-alkyl; Cw-alkyl, substituted io in which R is CM-alkyl or phenyl, these radicals optionally being substituted on a carbon atom by one or more substituents which are selected independently of one another from the following radicals: CM-alkyl; C 1-4 alkyl substituted by hydroxy, amino, carboxy by hydroxy, amino, carboxy or halogen; C ^ cycloalkyl; 15 or halogen; C ^ cycloalkyl; Cw alkoxy; CM alkylthio;

Cw alkoxy; Q ^ alkyltio; Amino; Cw alkyamino; Di (CW-alkyl) amino; Halogen; Cw alkanoylamino; Cw alkanoyloxy; Carboxy;

0

II

Amino; Cw alkyamino; Di (Cw alkyl) amino; CM-AlkanoyI-amino; Carboxy;

0

-C-OC. L-alkyl}

20 1

Hydroxy; Amidino; Guanidino; Trifluoromethyl; Phenyl; Phenyl substituted by one, two or three amino, halogen, hydroxy, trifluoromethyl, Cw-alkyl or C ^ alkoxy groups; and heteroaryl or heteroaralkyl, the heteroatom

-C-OC ^ -AUtyl;

Hydroxy; Amidino; Guanidino; Trifluoromethyl; Phenyl; Phenyl substituted by one, two or three amino, halogen,

Hydroxy, trifluoromethyl, C 1-4 alkyl or C 1-4 alkoxy groups; and heteroaryl or heteroaralkyl, the heteroatom 25 or the heteroatoms in the above-mentioned heterocyclic or the heteroatoms in the above-mentioned heterocyclic units being selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl unit 1 to 6 connected to the said heteroaralkyl unit Has carbon atoms or, if desired, are substituted so that a fused carbocyclic or heterocyclic ring is formed;

Units from 1 to 4 oxygen, nitrogen or sulfur atoms are selected and the alkyl unit connected to the heteroalkyl unit mentioned has 1 to 6 carbon atoms.

30 In the above subclass, the preferred compounds are those in which the substituent A is -CH2CH2-,

-chch2-,

(f)

ch,

r

• N - \\

35

-Ç ~ \ or -CH2ÇH- ^ CH,

x

N

or

N N-

l Ì

, and wherein 40 (a) R1 and R8 together represent

Ü0CHo c =

45

where X is O, S or NR, where R Q_r is alkyl or phenyl, these radicals being optionally substituted on a carbon atom by one or more substituents which are independently selected from the following radicals: Cw-alkyl; Cw alkyl substituted by hydroxy, amino, carboxy or halogen; C ^ cycloalkyl; Ci_4-alkoxy; C ^ alkylthio; Amino; Cw alkylamino; Di (CW-50 alkyl) amino; Halogen; Cw alkanoylamino; C ^ alkanoyloxy; Carboxy;

0

CH,

stand or

(b) R8 represents a hydrogen atom and R1 represents a hydrogen atom, CH3CH2-,

CH,

ch-

ch.

oh ch

or

OH

ch3ch-

stands.

Compounds in which R8 is a what hydroxy; Amidino; Guanidino; trifluoromethyl; Phenyl; Phenyl, serstoffatom means and R1 for

-C-0C.J alkyl;

substituted by one, two or three amino, halogen, hydroxy, trifluoromethyl, Q_4 alkyl or Cw alkoxy groups; and heteroaryl or heteroaralkyl, the heteroatom or heteroatoms in the abovementioned heterocyclic units being selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl unit connected to said heteroaralkyl unit having 1 to 6 carbon atoms; and

(G)

n-

- N-

/ \ n <vn ~ r

-N ~

\

/

N ^ N-R

OH CH ^ CH-

60 stands, the preferred compounds having the absolute configuration 5R, 6S, 8R.

According to a particularly preferred embodiment according to the invention, compounds are obtained in which the rest of the formula

65 © ^ —v

-O

a remainder of the formula

658 248 10

e (a) R1 and R8 together for

—K / '^ l 7

—R h0ch2

c =

5 «r 7 5 ch ^

means in which R, R and R independently of one another a 3rd

Is a hydrogen atom, a Cw-alkyl group, a C ^ -alkoxy or group, a CM-alkyl group, substituted by a hydroxy- (b) R8 is a hydrogen atom and R1 for hydrogen, group, a C ^ -alkylthio group, an amino- , Carboxy and CH3H2,

Mean carbamoyl group. In this subclass, the 10th

preferred compounds are those in which the substituent A for v3 i i

-CH2CH2, ck-, ^ c- or ch3ch-

-chch2-, ch3 ch3

CH3 15 stands.

_ / \ or _CH CH_ Particularly preferred are compounds in which R8 is

> - '2' is hydrogen and R1 is

CHï

3 OH

and where either “„ •

(a) R1 and R8 together for 20 3

H0CH stands. The preferred compounds have the absolute confi-

guration 5R, 6S, 8R.

c = a further preferred embodiment according to the invention

CH, 25 form comprises the preparation of compounds in which the rest of the general formula are or

(b) R8 represents a hydrogen atom and R1 represents hydrogen,

CH3CH2,

® -—x

■ o ch, ch, oh oh 30.

\ 3 | | a radical of the general formula ch-, _ c- or ch3ch-

ch3 CH3 —sch3

stands. n ^ / - ^ '~ ch3

Compounds in which R8 is 35 are particularly preferred

Means hydrogen atom and R1 means for.

OH In this subclass, preferred compounds are those

CHjCH- wherein the substituent A for

4fi —chch -

stands. The preferred compounds have the absolute confi, 2 ~ »

guration 5R, 6S, 8R. c%

A most preferred embodiment according to the invention comprises the preparation of compounds in which the -CH2CH2-,

Rest of the general formula

0

-O

45

or -ch2ch-

ch3

is a radical of the general formula and wherein either

, 6 (a) R1 and R8 together for

71 —R

high-

CH3

means in which R5, R6 and R7 are independently a 55 or

Hydrogen atom, a C ^ alkyl, Cw alkoxy, through a (b) R8 represents a hydrogen atom and R1 for hydrogen

Hydroxy group substituted CM alkyl, C ^ alkylthio and ch, ch, oh

Amino group mean. In this subclass are the preferred v3 ^ 3

preferred compounds are those in which the substituent A for ch, ch? -, ch- \ c-

-ch2CH2-, 60 c-r - "*

. -CHCHo-f

CH3 or

- / \ or -CHoC

H

CH3

ch3 oh ch, ch-

or -choch- 65

CH3 stands.

Particularly preferred are compounds in which R8 is and where either is hydrogen and R1 is

11

oh ch ^ ch-

658 248

sh

(7) -chjchj n stands. The preferred connections have the absolute configuration 5R, 6S, 8R.

Another preferred embodiment according to the invention comprises the preparation of compounds in which the rest of the general formula

-O

represents a pyridinium radical. In this subclass, preferred compounds are those in which the substituent A is chch2-, -ch2ch2- or -ch2ch-

JL »» s ntr

(9)

■ Œ2Œ2

ch20h (8) -ch, ch, en och,

'' \ N

2 2 h

... Svi V

10th

(U) -PCH2SC3

ch3 n— '

R or S dlastereoisomers

NH,

(10) -CHjCHj N

ck,

(12) -ca2ca2 N

\ '/ W

sh

CH ^

and in which either (a) R1 and R8 together represent highly

ch,

15 (13) -ch2ch28n /

20th

or (14)

X ^ - ^ 3

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

, c =

ch-

or and in which R2 represents a hydrogen atom, an anionic charge or a customary, easily removable carboxyl protective group, with the proviso that if R2 represents a hydrogen atom or a protective group, a counter anion is also present,

(b) R8 represents a hydrogen atom and R1 represents hydrogen, and the pharmaceutically acceptable acid addition salts

CH3CH2,

ch ch.

oh ch-

ch.

30th

from that.

Another preferred class of quaternized R14 substituents are those of the general formula

16

r "

or in which R16 is selected from the following substituted and 35 unsubstituted radicals: alkyl, alkenyl and alkynyl having 1 to 10 carbon atoms; Cycloalkyl and cycloalkylalkyl is 3 to 6. Carbon atoms in the cycloalkyl ring and 1 to 6 carbon

Compounds in which R8 is a was atom in the alkyl units are particularly preferred; Phenyl; Aralkyl, aralkenyl and seratomatom means and R1 for aralkynyl, in which the aryl unit is phenyl and the aliphatic

40 parts has 1 to 6 carbon atoms; Heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, where the heteroatom or the heteroatoms is in the above-mentioned heterocyclic. The preferred compounds have the absolute confi units under 1 to 4 oxygen, nitrogen or sulfur 5G, 6S, 8R configuration. Men are selected and those with the heterocyclic

A most preferred embodiment according to the invention of 45 units of linked alkyl units of 1 to 6 carbon atoms comprises the preparation of compounds of the general; where the above-mentioned radicals R16, if desired, oh

oh yeah

my formula oh h

0 "- coor wherein the rest of the general formula

®.—, -a-nj has the following meanings:

are substituted by 1 to 3 substituents which are independently selected from the following radicals: ~ CW) -alkyl, optionally substituted by amino, 50 fluorine, chlorine, carboxyl, hydroxy or carbamoyl;

Fluorine, chlorine or bromine;

-OR3; -0C02R3; -OCOR3; -OCONR3R4; -0S02R3; -Oxo; -NR3R4; R3CONR4-; -NR3C02R4; -NR3CONR3R4; -NR3S02R4; -SR3;

55 O

f Q

-s-r *;

ch,

(l) -ch2ck2 "n

(2) -ch2ch2 n)

't

0 0 (s7> -s-r

60

(3) -CH2CH2eN / ^

.ch.

(4) -CE2CH2-M

.ch 2 OS

'l \\

H,

(5) -CK ^ CH.6! ^ ^) ~ CH3

(6) -CH2CH2

-SO3R3; -C02R3; -CONR3R4; -CN; or a phenyl radical which is optionally substituted by: 1-3-fluorine, chlorine, bromine, C 1-4 alkyl, -OR3, -NR3R4, -S03R3, -C02R3 or -CONR3R4, where R3, R4 and R9 are ducks in these substituents Rî6 have the meanings given above; j—, or in what

ä, v> -sch, R16 is a divalent phenylene or Cw alkylene group, which with the

\ /

658 248

12

Ring is connected so that a bridged, polycyclic group forms, and the rest of the general formula

-KD

means a substituted or unsubstituted, mono-, bi- or poly-cyclic, non-aromatic, heterocyclic radical which can be fused to another aromatic or non-aromatic ring and which has at least one nitrogen atom in the ring and is bonded to A via a ring nitrogen atom is, which results in a quaternary ammonium group. The heterocyclic radical can be saturated or unsaturated (with 1 to 2 double bonds) and can contain up to two further heteroatoms in addition to the quaternary nitrogen atom. Such additional heteroatoms are O, S (0) m, N, NR15 or NR17R18, in which m represents 0.1 or 2, R15 represents a hydrogen atom, an optionally substituted Ci_6-alkyl or optionally substituted phenyl group and R17 and R18 independently of one another mean a optionally substituted Ci - ^ - alkyl or optionally substituted phenyl group.

In a preferred embodiment, the rest of the general formula denotes a non-aromatic, 4- to 7-membered, preferably 5- or O-membered, N-containing heterocyclic ring with 0 to 2 double bonds and 0 to 2 additional heteroatoms, which are listed under O, S (0) m, N, NR15 or NR17R18 are selected, in which m stands for 0.1 or 2, RI5 for a hydrogen atom, a CMAalkyl group which is optionally substituted by 1 to 2 substituents, these substituents being selected independently of one another under-OR3 , -NR3R4, -C02R3, oxo, phenyl, fluorine, chlorine, bromine, -S03R3 and -CONR3R4, or phenyl group, which is optionally substituted by 1 to 3 substituents, these substituents being selected independently of one another from CM-alkyl-, - OR3, -NR3R4, fluorine, chlorine, bromine, -S03R3, -C02R2 or -CONR3R4, and R17 and R18 each independently represent a Q ^ -alkyl group, which is optionally substituted by 1 or 2 substituents i these substituents are independently selected from -OR3, -NR3R4, -C02R3, oxo, phenyl, fluorine, chlorine, bromine, -S03R3 and -CONR3R4, or a phenyl group which is optionally substituted by 1 to 3 substituents, whereby these substituents are selected independently of one another from CM-alkyl, -OR3, -NR3R4, fluorine, chlorine, bromine, -S03R3, -C02R2 and -CONR3R4, where R3 and R4 in these heterocyclic NR15 or NR17R18 groups have the above in connection with have the meanings given for R16. In such a preferred embodiment, the ring

—10

if desired, be substituted by 1 to 3 of the following substituents:

(a) C 1-4 alkyl, optionally substituted by 1 to 2 substituents which are selected independently of one another from fluorine, chlorine, bromine, -OR3, -OCOR3, -OCONR3R4, oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -NR3S02R4 , -SR3, -S03R \ -CO, R3, -C02R3 and -CONR3R4;

(b) CM-alkenyl, optionally substituted by 1 to 2 substituents which are selected independently of one another from fluorine, chlorine, bromine, -OR3, -OCOR3, -OCONR3R4, oxo,

-NR3R4, -NR3R4, -NR3COR4, -NR3CONR3R4, -NR3SO, R4 ,. -SR3, -S03R3, -CO, R3 and -CONR3R4;

(c) C2_o-alkynyl, optionally substituted by 1 to 2 substituents which are selected independently of one another from fluorine, chlorine, bromine, -OR3, -OCOR3, -OCONR3R4, oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -NR3SO, R4, -SR3, -S03R3 and -CONR3R4;

(d) C 1 -C -cycloalkyl, optionally substituted by 1 to 2 substituents which are selected independently of one another from fluorine, chlorine, bromine -OR3, -OCOR3, -OCONR3R4, oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -NR3S02R4, -SR3, -SO3R3, -C02R3 and -CONR3R4;

(e) Cycloalkylalkyl having 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl unit, optionally substituted by 1 to 2 substituents which are selected independently of one another from fluorine, chlorine, bromine, -OR3, -OCOR3, -OCONR3R4, oxo , -NR3R4, -NR3COR4, -NR3CONR3R4, -NR3S02R \ -SR3, -S03R3, -C02R3 and CONR3R4;

(f) Heteroaryl, where the heteroatom or the heteroatoms are selected from 1 to 4 oxygen, nitrogen or sulfur atoms, optionally substituted by 1 to 2 substituents, which are selected independently of one another from fluorine, chlorine, bromine, -OR3, -OCOR3 , -OCONR3R4, Oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -NR3S02R4, -SR3, -S03R \ -C02R3 and -CONR3R4; preferred heteroaryl radicals are 5- or 6-membered, aromatic, heterocyclic rings;

(g) Heteroaralkyl, where the heteroatom or the heteroatoms are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl unit has 1 to 6 carbon atoms, optionally substituted by 1 or 2 substituents which are selected independently of one another from fluorine, chlorine , Bromine, -OR3, -OCOR3, -OCONR3R \ Oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -NR3S02R4, -SR3, -S03R3, -C02R3 and -CONR3R4; preferred heteroaralkyl groups are those in which the herteroaryl radical is a 5- or o-membered aromatic, heterocyclic ring and the alkyl unit has 1 to 2 carbon atoms;

(h) Heterocyclyl, the hetero atom or the hetero atoms being selected from 1 to 4 oxygen, nitrogen or sulfur atoms, optionally substituted by 1 to 2 substituents which are selected independently of one another from fluorine, chlorine, bromine, -OR3, -OCOR3 , -OCONR3R4, Oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -NR3SO2R4, -SR3, -SO3R3, -C02R3 and -CONR3R4; preferred herterocyclyl groups are 5- or 6-membered, saturated or unsaturated rings;

(i) Heterocyclylalkyl, where the heteroatom or the heteroatoms are selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl unit has 1 to 6 carbon atoms, optionally substituted by 1 to 2 substituents which are selected independently of one another from fluorine, chlorine, bromine , -OR3, -OCOR3, -OCONR3R4, Oxo,

-nr3r4, -nr3cor4, -nr3conr3r4, -nr3so2r4, -sr3, -so3r3,

-C02R3 and -CONR3R4; preferred heterocyclylalkyl radicals are those in which the heterocyclyl unit is a 5- or membered saturated or unsaturated ring.

(j) fluorine, chlorine or bromine;

(k) -OR3;

(1) -0C02R3;

(m) -OCÖR3;

(n) -OCONR3R4;

(o) -0S02R3;

(p) oxo;

(q) -NR3R4;

(r) R3CONR4;

(s) -nr3coor4;

(t) -NR3CONR3R4;

5

10th

15

20th

25th

30th

35

40

45

50

55

60

65

13

658 248

(u) -NR3S02R4;

(v) -SR3;

O

î

(w) -S-R9;

O O

v,

(x) -S-R9;

(y) -SO3R3;

(z) -C02R3;

(aa) -CONR3R4;

(bb) -CN; or

(cc) phenyl, optionally substituted by 1-3 fluorine, chlorine, bromine, CM-alkyl, -OR3, -NR3R4, -SO3R3, -C02R3 or -CONR3R4.

The substituents R3, R4 and R9 given above have the meanings given in connection with the substituents R1.

The ring defined above

is a non-aromatic, heterocyclic group. However, this ring can be fused to another ring, which is a saturated or unsaturated carbocyclic ring, preferably a C4_7-carbocyclic ring, a phenyl ring, a 4- to 7-membered heterocyclic (saturated or unsaturated) ring with 1 to 3 heteroatoms , which are selected from O, N, S (0) m, NR15 and NRnR18, or a 5- to 8-membered heteroaromatic ring which contains 1 to 3 heteroatoms, which are selected from O, S (0) m, N, NR1Sund NR17R1S are selected, wherein m, R15, R17 and R18 have the meanings given above.

The substituent R16 of the non-aromatic radical R14 can either (a) be a optionally substituted CM-alkyl-, Q-io-alkenyl, Qk, -alkynyl-, C ^, - cycloalkyl-, C ^ -cycloalkyl-C ^ - alkyl-, phenyl-, phenyl-Cw-alkyl-, phenyl-C ^ -alkenyl-, phenyl-C2-6-alkynyl-, heteroaryl-, heteroaralkyl-, in which the alkyl unit has 1 to 6 carbon atoms, heterocyclyl- or heterocyclylalkyl- Mean group in which the alkyl unit has 1 to 6 carbon atoms, or (b) mean a divalent phenylene group or C 1-4 alkylene group attached to the ring

- N

is bound so that a ring-bridged polycyclic group is formed, e.g. B. a quinuclide group. The heteroaryl substituent (or the heteroaryl part of the heteroaralkyl substituent) can be a mono-, bi- or polycyclic, aromatic, heterocyclic group which contains 1 to 4 O, N or S atoms; preferred are 5- or o-membered heterocyclic rings such as thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, tetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyrylazolyl, pyridazinyl, pyridazinyl. The heterocyclyl substituent (or the heterocyclyl part of the heterocyclylalkyl substituent) can be a mono-, bi- or polycyclic, saturated or unsaturated, non-aromatic, heterocyclic group containing 1 to 4 O, N or S atoms; 5- or o-membered heterocyclic rings, such as morpholinyl, piperazinyl, piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrrolinyl and pyrrolidinyl, are preferred.

If the substituent R16 represents an alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, phenyl, phenylalkyl, phenylalkenyl, phenylalkynyl, heteroaryl, heteroaralkyl, heterocyclyl or heterocyclylalkyl group, these can be used

If desired, groups can be substituted by 1 to 3 substituents which are selected from the following radicals;

(a) CM-alkyl, optionally substituted by preferably 1 to 3 amino, fluorine, chlorine, carboxyl, hydroxyl or 5 carbamoyl groups;

(b) fluorine, chlorine or bromine;

(c) -OR3;

(d) -0C02R3;

(e) -OCOR3;

10 (f) -OCONR3R4;

(g) -0S02R3;

(h) oxo;

(i) -NR3R4;

(j) R3CONR4;

15 (k) -NR3C02R4;

(1) -NR3CONR3R4;

(m) -NR3S02R4;

(n) -SR3;

(0) -SOR9;

20 (p) -S02R9;

(q) -SO3R3;

(r) -C02R3;

(s) -CONR3R4;

(t) -CN; or

25 (u) phenyl, optionally substituted by 1 to 3 substituents which are independently selected from fluorine, chlorine, bromine, C 1-4 alkyl, -OR3, -NR3R \ -S03R3, -C02R3 or CNR3R4, in the above substituents R16 the groups R3 and R4 are independently selected from water-30 substance; Alkyl, alkenyl and alkynyl of 1 to 10 carbon atoms; Cycloalkyl, cycloalkylalkyl and alkylcycloalkyl having 3 to 6 carbon atoms in the cycloalkyl ring having 1 to 6 carbon atoms in the alkyl units; Phenyl; Aralkyl, aralkenyl and aralkynyl, the aryl unit being phenyl and the aliphatic part having 1 to 6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, where the heteroaryl and heterocyclyl groups or part of this group is as previously defined for R16 and the alkyl units connected to said heterocyclic units have 1 to 6 40 carbon atoms; or R3 and R4, together with the nitrogen atom to which at least one of these radicals is bonded, can form a 5-membered or membered nitrogen-containing heterocyclic ring (as defined above for R5); and R9 has the meanings given above for R3, but cannot stand for hydrogen. The most preferred substituent R16 is a CWI alkyl group, especially a methyl group.

If R16 represents a divalent phenylene or Cw alkylene group, then this group is bonded to another atom of ring 50 of the formula, so that a bridged, polycyclic ring 55 is formed, e.g. B. a quaternized quinuclidine ring of the formula

CD

K,

According to a particularly preferred embodiment, compounds of the general formula I are prepared according to the invention, in which the rest of the formula

, 16th

65

o has the following meanings:

658 248

14

ch "\

ch3

ch "

ch3 r ch.

ch

—N9 S -

V /

ch

■ n® /

w nh /

eV

A.

\ ch / \

s- *},

n-ch.

V /

~ \ _y ch ,, '\

_ \ l 'N

ch

-ch

10th

ru

<3T \

ch ch

-êN

ch

/ -CH3 '"S1? S_> °

\ /

y

-A

s-

v_y

/ ■

—N

3rd

—N "

A

CÏ3 / TA • /

N

ch,

v_y '\

/ H or -

v_y

CK 15 In this preferred subclass, preferred Verbin-3 fertilize those in which A is ch,

-chch,

i £

CH,

£> ■

-chgch-ch,

or

—N

20th

- (CH2) n-, where n is 2, 3 or 4, more preferably those in which A is -CH2CH2-, -CH2CH2CH2-,

25th

-chch2

ch,

-P

wherein Y is a hydrogen, chlorine, bromine, iodine or fluorine atom or a C ^ alkyl, hydroxy, -SCM alkyl, carboxyl, carb

means amoyl or phenyl group. In this subclass are "" 3

and most preferably those in which A is -CH2CH2-and in which either or -ch2ch-ch,

preferred compounds are those in which A is

-chch2-, - / ~ \, -ch2ch-ch, '™

(a) R1 and R8 together for ch,

3 "" 3

or - (CH2) n stands,

where n is 2, 3 or 4, and in particular those in which A is -CH2CH2-, -CH2CH2CH2-,

-chch2-, -çy or -ch2ch-

30th

high,

ch,

c =

ch.

ch,

, and most preferably those wherein A is -CH2CH2- and where either (a) R1 and R8 together are or

35 (b) R8 represents a hydrogen atom and R1 represents hydrogen, CH3CH2-,

ch, ch, oh oh

3 \ 3- i «

ch-, yc or ch, ch-

40

ch,

ch,

high-,

ch,

45

stands.

Particularly preferred are compounds in which R8 represents a hydrogen atom and R1 for

OH

ch3ch-

stand or

(b) R8 is hydrogen and R1 is hydrogen, CH3CH2, in particular compounds of the absolute configuration

5R, 6S, 8R.

ch,

ch,

ch-

ch, oh 3 ^ ■

ch,

oh c- or ch ^ ch-

In a further, more preferred embodiment 50, compounds of the general formula I are prepared according to the invention, in which the rest of the formula is.

Particularly preferred are compounds in which R8 is a hydrogen atom and R1 is 55

, 16th

-N

oh yeah

for ch,

X3-

stands, in particular compounds of the absolute configuration 5R, 6S, 8R are preferred. 60

In an even more preferred embodiment, compounds of the general formula I are prepared according to the invention in which Y is hydrogen, chlorine, bromine or iodine or in which the rest of the general formula is fluorine atom or a CM-alkyl, hydroxy, -S-CM-alkyl-,

s 16 means carboxyl, carbamoyl or phenyl group. In this

65 preferred subclass are the preferred compounds those in which A is - (CH2) n-, where n is 2, 3 or 4. Most preferred are those compounds in which A has the following meanings: -CH2CH2- and in which either

\.

O

15

658 248

(a) R1 and R8 together for high2

ch,

wherein

"C =

For

\ s.

"~ 6 ^

5 C? 3

r stand or

(b) R8 represents a hydrogen atom and R1 represents hydrogen, CH3CH2-,

fV

Y% /

ch

'V

0

y

\

S ^ -0

CH,

ch,

ch-

ch, oh 5

oh c- or ch, ch-

10th

(both a- and ß-

Diaster soisoners)

ch ch

ch.

stands.

Particularly preferred are compounds in which R8 is a hydrogen atom and R1 is ohch-

stands, in particular connections of the absolute configuration 5R, 6S, 8R.

In a most preferred embodiment according to the invention

15

V

N-CH. or y j

\ 6 / CH3

.X Li w

and in which R2 represents a hydrogen atom, an anionic charge or a conventional, easily removable carboxyl protective group, with the proviso that if R2 represents a hydrogen atom or a protective group, a counterion is also present 20 and the pharmaceutically acceptable addition salts thereof. Certain compounds falling under the general formula I can exist both as optical isomers and as epimer mixtures. According to the invention, all of these management forms are compounds of the general formula optical isomers and the epimer mixtures. Han-I, in which the rest of the formula 25 is, for example, the substituent in the 6-position

16 ^^ a hydroxyethyl group, this substituent can be in both the R and S configuration. Thus, according to the invention, both the isomers obtained and their epimer mixtures are included.

for 30 In the process according to the invention, an intermediate

"MD

compound of the general formula IV

R8 H

pi stands. 35

In this preferred subclass, preferred compounds are those in which A is - (CH2) "-, where n is 2, 3 or 4, and most preferred are those in which A is -CH2CH2- and wherein either

(a) R1 and R8 together for f T

// 0

coor iv high,

. c =

ch,

stand or

(b) R8 represents a hydrogen atom and R1 represents hydrogen, CH3CH2-

ch,

ch,

ch-

ch, oh

D *

oh ch

, c- or chjch-

40, which is disclosed, for example, in European patent application 38 869 and which can be produced by the general processes described there. L denotes a customary leaving group (it is referred to as “X” in European patent application 38 869), such as chlorine, bromine, iodine, 45 benzenesulfonyloxy, p-toluenesulfonyloxy, p-nitrobenzenesulfonyloxy, methanesulfonyloxy, trifluoromethanesulfonyloxy, diphenoxyoxyphosphinyloxy or di - (trichloroethoxy) phosphinyloxy. The preferred leaving group is diphenoxyphosphinyloxy. The intermediate compounds of the general formula IV are generally formed in situ by reacting an intermediate compound of the general formula III

stands.

Particularly preferred are compounds in which R8 is a hydrogen atom and R1 is oh ch ^ ch-

stands, in particular connections of the absolute configuration 5R, 6S, 8R.

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

55

H

y

T

-COOR

s-ch, ch

COOR "

S

iii

60 wherein R1, R8 and R2 'have the meanings given above, with a suitable acylating agent R ° -L. The preferred intermediate IV, wherein L represents a diphenoxy-phosphinyloxy group, can be prepared by using a ketoester of formula III in an inert organic solvent such as methylene chloride, acetonitrile or dimethylformamide with about one mole equivalent of diphenyl chlorophosphate in the presence a base such as diisopropylethylamine, triethylamine, 4-dimethylaminopyridine or the like

658 248

16

a temperature of about -20 ° C to + 40 ° C, most preferably at about 0 ° C. If desired, the intermediate compound IV can be isolated; however, they are usually used without further isolation or purification as the starting compound for the process according to the invention.

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

HS-A-R14 VII

io

X®

wherein A represents a cyclopentylene, cyclohexylene or C2 ^ alkylene group, which is optionally substituted by one or more Cw-alkyl groups, most preferably for a cyclopentylene group, a cyclohexylene group or for

15

, 10th

-C-

t

.11

R12

f

-c-

r

»13

R '1 R'

stands in which R10, Rn, R12 and R13 each independently represent a hydrogen atom or a C ^ alkyl group, X® represents a counterion which is derived from a strong acid, for. B. Cl ", Br", CH3S03 ", CF3S03" or

CH-jSOJ

CF-SO ^ "

or CH-

r <0> "so3

20th

25th

30th

and R14 represents a quaternized, nitrogen-containing, aromatic or non-aromatic heterocycle as defined above. The reaction takes place in an inert solvent, such as acetonitrile, acetonitrile-dimethylformamide, tetrahydrofuran, tetrahydrofuran-H20, acetonitrile-H20 or acetone, in the presence of a base. The type of base is not critical. However, the 35 best results are obtained if a non-nucleophilic, tertiary amine is used as the base, such as diisopropylethylamine, 1,8-diazabicyclo [5.4.0] undec-7-ene, 1,5-diazabi-cyclo [ 4.3.0] non-5-enes, or a tri- (Cw) alkylamine such as triethylamine, tributylamine or tripopylamine. The reaction of the intermediate compound IV with the thiol VII can be carried out in a wide temperature range, e.g. B. at -15 ° C up to room temperature. It is preferred to operate in a temperature range from about -15 ° C to +15 ° C, most preferably at about 0 ° C.

The carbapenem compound obtained by reacting the quaternary aminthiol of formula VII with the intermediate compound IV has a counter anion [e.g. (C6H50) 2P02®, Cl®, or the quaternary thiol-associated anion] associated with the compound. This counter anion can be replaced by another counter anion at this stage by conventional methods, e.g. B. one that is pharmaceutically acceptable. Alternatively, the counter anion can be removed during the subsequent deblocking step. If the quaternized carbapenem compound and the counter anion form an insoluble product, the product can be allowed to crystallize out as it forms and can be obtained by filtering off.

After the desired carpabenem compound has been obtained, the carboxyl protective group R2 'of the 60 compound I' can, if desired, be removed by customary processes such as solvolysis, chemical reduction or hydrogenation. If the protective group is a p-nitrobenzyl, benzyl, benzhydryl or 2-naphthylmethyl group which can be easily removed by catalytic hydrogenation, the intermediate compound I 'is treated in a suitable solvent, such as dioxane-water-ethanol, Tetrahydrofuran-diethyl ether buffer, tetrahydrofuran-aqueous dipotassium hy-

45

50

55

drug phosphate isopropanol or the like, at a hydrogen pressure of 1 to 4 at in the presence of a hydrogenation catalyst such as palladium on carbon, palladium hydroxide, platinum oxide or the like, at a temperature of 0 to 50 ° C for about 0.24 to 4 Hours. If the group R2 'is o-nitrobenzyl, then photolysis can be used for deblocking. Protecting groups such as 2,2,2-trichloroethyl can be removed by mild reduction with zinc. The allyl protecting group can be removed using a catalyst such as a mixture of a palladium compound and triphenylphosphine in a suitable aprotic solvent such as tetrahydrofuran, methylene chloride or diethyl ether. Similarly, other conventional carboxyl protecting groups can be removed by methods known to those skilled in the art. Finally, as already stated above, the compounds of the formula I 'in which R2' is a physiologically hydrolyzable ester group, such as acetoxymethyl, phthalidyl, indanyl, pivaloyloxymethyl, methoxymethyl etc., can be administered directly to the host without deblocking, since such esters are hydrolyzed in vivo under physiological conditions.

If the substituent R1 and / or R8 or the quaternized nucleophile R14 bound to the substituent A contains a functional group which could interfere with the intended course of the reaction, this group can be protected with a customary blocking group and then deblocked to give the desired one regenerate functional group. Suitable blocking groups and methods for introducing and removing such groups are well known to those skilled in the art.

Like the other β-lactam antibiotics, the compounds of the general formula I can be converted into pharmaceutically acceptable salts by known processes, which for the purposes of the invention are essentially equivalent to the compounds which have not been converted into the salts. For example, a compound of the general formula I in which R 2 is an anionic charge can be dissolved in a suitable inert solvent. Then you add an equivalent of a pharmaceutically acceptable acid. The desired acid addition salt can be obtained by conventional methods, e.g. B. solvent precipitation, lyophilization, etc., win. If other basic or acidic functional groups are present in the compound of the general formula I, the pharmaceutically acceptable base addition salts and acid addition salts can be obtained in a similar manner by known processes.

The compounds of the general formula I, in which R2 denotes a hydrogen atom or an anionic charge, or pharmaceutically acceptable salts thereof, can also be converted into the corresponding compounds by conventional methods, in which R2 denotes a physiologically hydrolyzable ester group. Compounds of the general formula I in which R 2 is a customary carboxyl protecting group can be converted into the corresponding compounds in which R 2 is a hydrogen atom, an anionic charge or a physiologically hydrolyzable ester group or in the pharmaceutically acceptable salts thereof.

Some of the thiol intermediates of general formula VII can be prepared by, for example, a sulfide of the formula or

villa

65

S

-, 10 / \ P. -7-_

R11 S

\

12

VT IIb

Ville

17th

658 248

wherein R10, R11, R12 and R13 each independently represent a hydrogen atom or a Q ^ alkyl group, with a heteroaromatic amine (as defined above) of the general formula o

or with a non-aromatic, heterocyclic amine (as defined above) of the general formula

R16

X)

and reacted with a strong acid. The reaction can be carried out in the presence or absence of an inert organic solvent. This solvent is preferably a non-polar, organic solvent such as methylene chloride, benzene, xylene, toluene or the like. If the amine and sulfide reagents are liquids or if a solid amine is soluble in a liquid sulfide reagent, the reaction is preferably carried out without an additional solvent by.

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

The formation of the quaternary aminthio intermediate compound of formula VII can be carried out in a temperature range from about -20 ° C to about 100 ° C. Is preferably carried out at temperatures of about 50 to 70 ° C.

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

The quaternary aminthiol intermediate has an associated counter anion, which is derived from the acid used. It is of course possible at this point to replace the anion with another counter anion using conventional methods, which is then used in the subsequent reaction with the carbapenem intermediate IV.

The carbapenem derivatives of the general formula I, in which R2 denotes a hydrogen atom, an anionic charge or a physiologically hydrolyzable carboxyl protective group, or the pharmaceutically acceptable salts thereof, are effective antibiotics which can be used against various gram-positive and gram-negative bacteria. For example, they can also be used as animal feed additives to increase growth, as food preservatives, as bactericides for industrial applications, e.g. B. in water-based paints or in the washing water of paper mills to inhibit the growth of dangerous bacteria, and as a disinfectant to destroy or inhibit the growth of dangerous bacteria on medical and dental equipment. However, they are particularly useful in the treatment of infectious diseases caused by gram-negative or gram-positive bacteria in humans and animals.

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

The dose to be administered depends primarily on the compound used, the specially formulated composition, the mode of administration, the nature and the condition of the host. The dosage of course also depends on the specific site and the organism to be treated. The choice of the particularly preferred dosage and the application route 15 is left to the treating doctor. In general, however, the compounds are administered parenterally or orally to a mammal (human and animal) in an amount of about 5 to 200 mg / kg / day. Administration is generally in multiple doses, e.g. B. three or four times a day. 20 The following examples serve to illustrate the invention.

example 1

Preparation of 3- (2- (l-pyridinium) ethylthio) -6a- (l- (R) -25 hydroxyethyl) -7-oxo-l-azabicyclo [3.2.0] hept-2-ene-2-carboxylate A. l- (2-Mercaptoethyl) pyridinium methanesulfonate

30th

+ N

+ MsOH

55 C, 16 h

MsO

A suspension of pyridinium methanesulfonate in pyridine is prepared by dropwise adding methanesulfonic acid (1.95 ml, 0.03 mol) to pyridine (8.0 ml, 0.099 mol) while cooling. 35 Ethylene sulfide (1.96 ml, 0.033 mol) is added to this suspension. The mixture obtained is stirred at 55 ° C. for 16 h, concentrated to a thick syrup under reduced pressure, which is mixed with a few ml of water, and the solution is poured onto a column (40 × 16 cm) of (x-Bondapak C-18, which is eluted with water 40. Lyophilization of the appropriate fractions leads to a colorless syrup, yield 6.5 g (91%).

IR (film) vmax: 2300-2600 (br, SH), 1635 (pyridinium), 1490, 1200 (sulfonate), 1068.1060.1045.791, 780 cm "1

'H NMR (DMSO-d6) ô: 2.32 (3H, s, CH3SO3-), 2.61.2.70, 45 2.73.2.82 (IH, B part of the A2B system, SH), 3.07 (2H, m [with D20: 3.08 (2H, t, J = 6.5 Hz)], CH2S), 4.76 (2H, t, J = 6.5 Hz, CH2N + ), 8.19 (2H, m, Hm of pyridinium), 8.6 (IH, m, Ho of pyridinium), 9.18 (2H, dd, J = 6.8Hz, J = 1, 4Hz, Ho of Pyridinium)

50 UV (H20) Xmax: 206 (e 5230) 258 (e 3760) m | x.

Method A B. 1- (2-Mercaptoethyl) pyridinium chloride

MsO

_ Permutlt S-1

£ i ~

An aqueous solution of crude l- (2-mercaptoethyl) pyridi-60 nium methanesulfonate (9.4 g, 0.04 mol) is added to a column (2.5 x 41 cm) of Permutit S-1 CT. The column is eluted at a rate of 0.5 ml / min, the appropriate fractions are combined and lyophilized to give a yellow syrup, yield 7.0 g (100%), which is used for the next step 65.

! H NMR (D20) ò: 3.22 (2H, m, CH2S), 4.88 (m, CH, N +), 8.18 (2H, m, Hm from pyridinium), 8.7 (1H, m, Hp of pyridinium), 9.0 ppm (2H, m, Ho of pyridinium).

658 248

18th

Procedure B

A

Cl

Dp-nitrobenzyl-3- [2- (l-pyridinium) ethylthio] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2- carboxylate diphenyl phosphate

Pyridine hydrochloride (4.05 g, 35 mmol) and ethylene sulfide (2.1 ml, 35 mmol) are added to precooled (ice bath) pyridine (5.6 ml, 70 mmol). The mixture is warmed to 65 ° C. and stirred for 75 minutes, giving a two-phase system. The easier phase is removed. The remaining oil is washed with ether (5 x 10 ml) and a high vacuum is obtained, giving the title compound (90-100%), which is used for the next step.

C. p-nitrobenzyl-3- [2- (l-pyridinium) ethylthio] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2- en-2-carboxylate chloride

1) HKt (iI'r) 2

5 OH

'' XJ> °

0 '"| 3) HS COOPNB

1) NEt (iPr),

O *

2) ClP (OPh)? , n ^ SHsi

10th

4) NEt (lPr),

COOPNB

O

COOPNB 4) NEt (iPr) g

COOPNB

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

IR (Nujol) vmax: 3300 (OH), 1775 (C = 0 of β-lactam), 1690 (C = 0 of the PNB ester), 1630 (pyridinium), 1605 (phenyl of the PNB ester), 1515 (N02 ), 1335 cm "1

^ -NMR (DMSO-d6) ô: 1.17 (3H, d, J = 6, 1 Hz, CH3CHOH), 3.2-3.75 (5H, H-4, H-6, CH2S), 3 , 75-4.5 (2H, H-5, CH3CHOH), 4.92 (2H, br.t, J = 6.5 Hz, CH2N +), 5.18 (1H, d, J = 4.9 Hz , OH), 5.37 (center of ABq, yes b = 14.2 Hz, CH2 of PNB), 7.69 (2H, d, J = 8.7 Hz, Ho of PNB), 8.24 (d , J = 8.6 Hz, Hm from PNB), 8.0-8.4 (4H, Hm from PNB, Hm from pyridinium), 8.66 (1H, m, Hp from pyridinium), 9.17 (2H, br.d, J = 5.5 Hz, Ho of pyridinium).

The filtrate and the solvents used for washing are combined and diluted with 150 ml of ether. The supernatant liquid is decanted and the gummy product is dissolved in 40 ml of water containing enough acetonitrile to give a solution which is placed on a column (3 X10 cm) of [x-Bondapak C-18. The column is eluted with 10% acetonitrile-90% water (150 ml) and 50% acetonitrile-50% water (100 ml) mixtures. The appropriate fractions are combined and lyophilized after the acetonitrile has been removed in vacuo. A yellow powder is obtained. The NMR shows the presence of the title compound with some p-nitrobenzyl-3- [2- (l-pyridinium) ethylthio] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabi-cyclo [3.2.0] hept-2-en-2-carboxylate-diphenylphosphate (2: 1) is mixed. The powder is left in as little water as possible and passed over a column of 1.5 x 21 cm) of Permutit S-1 CL ~ with water. Lyophilization of the appropriate fractions gives 1.8 g (20%) of the title compound.

A solution of p-nitrobenzyl-6a- [l- (R) -hydroxyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2-carboxylate (0.174 g, 0.50 mmol) in acetonitrile ( 2 ml) is cooled to 0 ° C. under a nitrogen atmosphere, treated successively with diisopropylethyl-15-amine (0.105 ml, 0.60 mmol) and diphenyl chlorophosphate (0.124 ml, 0.60 mmol), the resulting solution is stirred at 0 ° C. for 30 min and treated sequentially with a solution of l- (2-mercap-toethyl) pyridinium methanesulfonate (0.170 g, 0.72 mmol) in 0.6 ml acetonitrile and diisopropylethylamine (0.105 ml, 20 0.60 mmol). The reaction mixture is stirred at 0 ° C for 15 min. Dilute with cold (0 ° C) water (7 ml) and pour the mixture onto a column (1.5x6.4 mm) of fi-Bondapak C-18. The column is eluted with a mixture of acetonitrile (25% -50%) in water (75% -50%). The appropriate fractions are combined and lyophilized after the acetonitrile has been removed in vacuo. A yellow powder is obtained; 0.33 g

(92%).

IR (KBr) vmax: 3600-3000 (OH), 1765 (C = 0 of ß-lactam), 1690 (C = 0 of the PNB ester), 1625 (pyridinium), 1585 (phenyl), 30 1510 (N02) , 1330 (N02), 885 cm "1 (N02)

'H NMR (DMSO-d6) ô: 1.16 (3H, d, J = 6.2 Hz, CH3CHOH), 4.87 (2H, br.t, J = 6.6 Hz, CH2S), 5 , 37 (center of ABq, Yes b = 14.3 Hz, CH2 from PNB), 6.7-7.5 (phenyl), 7.68 (d, J = 8.8 Hz, Ho from PNB), 8 , 23 (d, J = 8.8 Hz, Hm from PNB), 8.0-8.3 (m, Hm from pyridinium), 8.4-8.8 (1H, Hp from pyridinium), 9.09 (2H, dd, J = 6.7 Hz, J = 1.3 Hz, Ho of pyridinium).

25th

35

40

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

011

10 # Pd / C, H2

THF, ether

43 0 1 (PhO), PO ~

COOPNB

50

C00

To a solution of p-nitrobenzyl-3- [2- (l-pyridinium) ethylthio] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo- [3.2.0] hept-2 -en-2-carboxylate-diphenylphosphate (0.16 g, 55 0.22 mmol) in moist tetrahydrofuran (10 ml) are added ether (10 ml), potassium phosphate-monobasic sodium hydroxide buffer pH 7.4 (16 ml, 0 , 05 M) and 10% palladium-on-carbon (0.16 g). The resulting mixture is hydrogenated at 25 ° C and 2.76 bar (40 psi) for 1 hour. The two phases are separated and the 60 organic phase is washed with water (2x3 ml). The aqueous solutions are combined, washed with ether (2x 10 ml) and added to a column (1.5x6.2 cm) of [x-Bondapak C-18 after removing traces of the organic solvents in vacuo. The column is eluted with water and, after lyophilization of the 65 suitable fractions, a yellow powder is obtained, yield 0.062 g (84%).

IR (KBr) vmax: 3700-3000 (OH), 1755 (C = 0 of ß-lactam), 1630 (pyridinium), 1590 cm "1 (carboxylate)

19th

658 248

'H NMR (D20) ô: 1.22 (3H, d, J = 6.4 Hz, CH3CHOH), 2.92 (d, J = 9, 1 Hz, H-4), 2.97 (d , J = 9, 1 Hz, H-4), 3.20 (dd, J = 2.5 Hz, J = 6, 1 Hz, H-6), 3.44 (t, J = 6.0 Hz , CH2S), 3.93 (dd, J = 9, 1 Hz, J = 2.5 Hz, H-5), 4.82 (t, J = 6.0 Hz, CH2N +), 8.04 (m , Hm of pyridinium), 8.5 (m, Hp of pyridinium), 8.82 (dd, J = 3.2Hz, J = l, l Hz, Ho of pyridinium)

UV (H20) Xraax: 259 (e 5800), 296 (s 7030) m \ i Tiß = 13.5 h (measured at a concentration of 10 "4 M in phosphate buffer pH 7.4 at 36.8 ° C).

Procedure B

Oh

A

Cl

10% Pd / C, H2

THF, ether, buffer 7.2

Bondapak C-18. The column is eluted with water and the appropriate fractions are lyophilized to give a colorless syrup, yield 2.4 g (91%).

IR (film) vmax: 2520 (SH), 1628 (pyridinium), 1600.1495, s 1325.1305, 1283.1200 (sulfonate), 1040, 938, 765, 680 cm "1. ^ -NMR (DMSO-d6 ) ô: 2.31 (3H, s, CH3SO3-), 2.47 (6H, s, CH3 of pyridinium), 2.57.2.66.2.69.2.78 (1H, B part of the A2B-Systems, SH), 3.06 (2H, m [with added D20 (2H, t, J = 6.5 Hz)], CH? S), 4.65 (2H, t, J = 6.5 Hz, CH2N +), 8.34 (1H, s, io Hp of pyridinium), 8.79 (2H, s, Ho of pyridinium) UV (H20) lm, x: 271 (e 4860) m [x analysis: for C10H17N03S2-0.5H20 calculated: C 44.09% 'H 6.66% N 5.14% S 23.54% found: C 44.26% H 6.49% N 5.17% S 24.18%

15

coopnb

Bp-nitrobenzyl-3- [2- (1- (3,5-dimethylpyridinium)) ethylthio] -6a- [1- (R) -hydroxyethyl] -7-oxo-l-azabicyclo- [3.2.0] hept -2-en-2-carboxylate diphenyl phosphate

To a solution of p-nitrobenzyl-3- [2- (l-pyridinium) ethylthio] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo- [3.2.0] hept-2 -en-2-carboxylate chloride (5.77 g, 11.4 mmol) in potassium phosphate-monobasic sodium hydroxide buffer (170 ml, 0.2 M, pH 7.22), tetrahydrofuran (30 ml) is added,

Ether (30 ml) and 10% palladium-on-carbon (5.7 g), the resulting mixture was hydrogenated for 1 h at 22 ° C. and 2.76 bar (40 psi) and filtered through a pad of Celite. The pillow is washed with water (2x15 ml), the filtrate and the washing water are combined and diluted with ether (100 ml). The aqueous phase is separated off, washed with ether (3 × 100 ml) and poured onto a column (4.5 × 20 cm) of u-Bondapak C-18 after the organic solvents have been removed in vacuo. The column is eluted with water and then with a mixture of 1% acetonitrile in water. After lyophilization of the appropriate fractions, 2.48 g (65%) of the title compound is obtained as a yellow powder. The analytical data are identical to those for the compound produced according to method A.

Example 2

Preparation of 3- [2- (l- (3,5-dimethylpyridinium) ethylthiol] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-ene -2-carboxylate

COOPNB

25th

4) NEt (iPr) 2

COOPNB

0

(PhO), &) ~

30th

COO

A. l- (2-Mercaptoethyl) -3,5-dimethylpyridinium methansulfonate

To a cold (0 ° C) solution of p-nitrobenzyl-6a- (l- (R) -hydroxyethyl) -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2-carboxylate (0.523 g, 1 , 50 mmol) in acetonitrile (6.0 ml) are added diisopropylethylamine (0.314 ml,

1.8 mmol) and then diphenyl chlorophosphate (0.373 ml, 1.8 mmol). The reaction mixture is stirred for 30 min and treated with a solution of l- (2-mercaptoethyl) -3,5-dimethylpyridinium methanesulfonate (0.493 g, 1.87 mmol) in acetonitrile (1.9 ml) and then with diisopropylethylamine (0.314 ml, 1.8 mmol). The reaction mixture is stirred for 1 h at 0 ° C, diluted with cold (0 ° C) water (26 ml) and poured onto a column (7.0 x 3.5 cm) of fi-Bondapak C-8. The column is eluted with a 25-50% acetonitrile-75-50% water mixture and after lyophilization of the appropriate fractions, 1.01 g (90%) of the title compound is obtained as a yellow powder.

IR (KBr) vmax: 3700-3100 (OH), 1778 (C = 0 of β-lactam), 1700 (C = O of the PNB ester), 1635 (pyridinium), 1595 (phenyl), 1521 (N02), 1335 (N02), 895 cm "1

"H NMR (DMSO-d6) Ô: 1.16 (3H, d, J = 6.1 Hz, CH3CHOH), 2.43 (s, CH3 of pyridinium), 4.75 (2H, m, CH2N +) , 5.38 45 (center of ABq, Yes b = 14.3 Hz, CH2 of PNB), 6.6-7.5 (10H, m, phenyl), 7.70 (2H, d, J = 8, 7 Hz, Ho from PNB), 8.0-8.5 (3H, m, Hp from pyridinium, Hm from PNB), 8.82 (2H, s, Ho from pyridinium)

UV (H20)> .max: 270 (e 11570), 306 (e 7343) my, 50 analysis for C37H38N3O10SP-H2O

calculated: C 58.03%, H 5.26%, N 5.48%, S 4.18% found: C 57.98%, H 5.05%, N 5.22%, S 4.34%

40

55

+ HaOH

MsO

C. 3- [2- (l- (3,5-Dimethylpyridinium)) ethylthio] -6a- [l (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2 -en-2-carboxylate

1034 Pd / c, h2

To a suspension of 3,5-lutidinium methanesulfonate in 3,5-lutidine, prepared by adding methanesulfonic acid (0.65 ml, 0.010 mol) to cold 3,5-lutidine (2.51 ml, 0.022 mol) ethylene sulfide (0.655 ml, 0.011 mol), the mixture obtained is stirred under a nitrogen atmosphere at 55 ° C. for 24 h, cooled to 23 ° C. and diluted with water (5 ml) and ether (5 ml). The organic layer is separated off and the aqueous solution is washed with ether (6 × 4 ml). The traces of ether are removed in vacuo and the solution is placed on a column (2.5 × 6.0 cm) of lieo

THF, ether,

buffer

COOPNB

(PhO), PO ~ «Wed

65

COO

To a solution of p-nitrobenzyl-3- [2- (l- (3,5-dimethylpyridinium)) ethylthio] -6a- [l- (R) -hydroxyethyl) -7-oxo-l-azabicy-

658 248

20th

clo [3.2.0] hept-2-en-2-carboxylate-diphenylphosphate (0.600 g, 0.80 mmol) in moist tetrahydrofuran (36 ml) are added ether (36 ml), potassium phosphate-monobasic sodium hydroxide buffer (0, 05 M, pH 7.4.44 ml) and 10% palladium-on-carbon (0.60 g), the resulting mixture is hydrogenated for 1.25 h at 23 ° C. and 2.76 bar (40 psi), the organic is separated Layer off and extracted with the buffer (2x5 ml). The water layers were combined, filtered through a pad of Celite, washed with ether (40 ml) and removed traces of the organic solvents in vacuo. Then the solution is placed on a column (2.5 x 10.0 cm) of liES ondapak C-18. The column is eluted with water and the appropriate fractions are lyophilized to give 0.186 g (64%) of the title compound as a yellowish powder.

IR (KBr) vmax: 3700-3100 (OH), 1760 (C = 0 of β-lactam), 1595 cm "1 (carboxylate)

* H-NMR (D20) ô: 1.21 (3H, d, J = 6.3 Hz, CH3CHOH), 2.45 (6H, s, CH3 on pyridinium), 2.81 (d, J = 9, 2Hz, H-4), 2.96 (d, J = 9.2 Hz, H-4), 3.22 (dd, J = 2.6 Hz, J = 2.6 Hz, H-6), 3.40 (t, J = 6.2Hz, CH, S), 3.84 (dd, J = 9.2Hz, J = 2.6 Hz, H-5), 4.15 (m, CH3CHOH), 4.71 (t, J = 6.2 Hz, CH, N +), 8.21 (1H, s, Hp of pyridinium), 8.46 (2H, s, Ho of pyridinium)

UV (H20) Xmax: 279 (s 8345), 296 (e 7714) mjx [«] § = +40.7 (c 0.53, H20)

ti = 16.9 h (measured at a concentration of 10 "4 in phosphate buffer pH 7.4 at 36.8 ° C).

Example 3

Preparation of (5R, 6S) -3 - [[2- (3-hydroxymethylpyridinio) ethyl] thio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo- [3.2.0 ] hept-2-en-2-carboxylate oh

A,

Oh

ch2oh n>

»^

co2pnb

10th

CO,

c02pnb ^^ 2p0

To a solution of p-nitrobenzyl- (5R, 6S) -6- [l- (R) -hydro-15 xyethyl [-3,7-dioxo-l-azabicyclo [3.2.0] heptane-2-carboxylate (0.174 g, 0.50 mmol) in 2 ml dry acetonitrile are added at 0 ° C under N2 diisopropylethylamine (0.096 ml, 0.55 mmol), diphenyl chlorophosphate (0.114 ml, 0.55 mmol) is added dropwise and the reaction mixture is stirred 30 min at 0 ° C. A solution 20 of 3-hydroxymethyl-l- (2-mercaptoethyl) pyridinium trifluoromethanesulfonate (0.223 g, 0.70 mmol) in 0.50 ml acetonitrile is then added, followed by diisopropylethylamine, (0.122 ml, 0.0 70 mmol), the mixture is kept at 0 ° C. for 30 min and then the reaction mixture is concentrated in vacuo. The remaining, yellow, gummy residue is taken up in water, adding enough acetonitrile that the gummy product goes into solution. This solution is placed on a C18 reverse phase column which is eluted with 15% acetonitrile water. The appropriate fractions are lyophilized and the 30 product (0.305 g; 81%) is obtained as a beige solid.

IR (KBr) v max: 3420 (br, OH), 1775 (β-lactam CO), 1695 (-C02PNB) CM "1

^ -NMR (d6-acetone) ô: 9.44-7.72 (m, 8H, aromatic), 7.22-6.91 (m, ÎOH, diphenylphosphate), 5.53.5.27 (ABq, J = 14 "Hz, 2H, benzylic), 5.04 (t, J = 7.4Hz, 2H, N-CH2), 4.75 (s, 2H, CH2OH), 4.5-3.1 (m , 8H), 1.21 (d, J = 6.3 Hz, 2H, CHMe).

25th

35

A. 3-hydroxymethyl-l- (2-mercaptoethyl) pyridinium trifluoromethanesulfonate ch-oh

N ()) + CFJSOJH +

CF3SO3-

40

C. (5R, 6S) -3- [2-hydroxymethylpyridinio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2 carboxylate oh

CHoOH

A

Trifluoromethanesulfonic acid (1.327 ml, 0.015 mol) is added dropwise to 3-pyridinemethanol (2.91 ml, 0.030 mol) and then ethylene sulfide (0.89 ml, 0.015 mol) is added. The homogeneous mixture obtained is heated (oil bath) for 20 h under N2 at 50 to 70 ° C, the reaction mixture is then taken up in water (15 ml) and extracted with CH2C12 (5x5 ml). The aqueous phase is concentrated in vacuo and then put it on a C] 8 reverse phase column. It is eluted with water and then the appropriate fractions are concentrated to give a pale yellow oil. This material is chromatographed again to give an almost colorless oil. After drying in vacuo (P2Os), the product (4.50 g; 94%) is obtained as a viscous oil.

IR (film) vmax: 3450 (st, OH), 2560 (schw, SH) CM "1! H-NMR (d6-acetone) ô: 9.10-8.05 (m, 4H, aromatic), 5, 01 (t, J = 5.5 Hz, 2H, N-CH,), 4.93 (s, 2H, -CH, OH), 4.43 br.s, 1H, -OH), 3.43- 3.18 (m, 2H, S-CH2), 2.34-2.10 (m, 1H, SH).

B. p-Nitrobenzyl- (5R, 6S) -3- [2- (3-hydroxymethylpyridinio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo- [3.2.0 ] hept-2-en-2-carboxylate diphenyl phosphate

"N

45

0

«To) 2Po ~

Oh

CHoOH

50

co, "

To a solution of p-nitrobenzyl- (5R, 6S) -3- [2- (3-hydroxy-methylpyridinio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-aza-bicyclo [ 3.2.0] hept-2-en-2-carboxylate-diphenylphosphate (0.145 g, 55 0.194 mmol) in 10 ml THF containing 5 drops of H20 are added 6.0 ml phosphate buffer (0.05 M, pH 7, 4), 0.145 g 10% palladium-on-carbon and 10 ml ether. The mixture is hydrogenated (Parr) for 1 h at 2.76 bar (40 psi), then filtered through a pad of Celite, the filter cake is washed with a little H20 and ether, 60 the aqueous phase is separated off and extracted three times with ether. The aqueous solution is then cooled to 0 ° C. and the pH is adjusted to 7.0 with pH 7.4 buffer. After the remaining volatile constituents have been removed in vacuo, the aqueous solution is added to a C18 reverse phase column, which is eluted with 65 water. Lyophilization of the appropriate fractions gives the product (36 mg, 51%) as a light yellow solid. Further purification by reverse phase HPLC gives the pure product (31 mg, 41%) as a solid.

21st

658 248

IR (KBr) v max: 3300 (br, OH), 1755 (β-lactam CO), 1590 (-COf) cm "1

'H NMR (D20) ô: 8.78-7.94 (m, 4H, aromatic), 4.83 (t, J = 6.0 Hz, 2H, N-CH2), 4.83 (s, 2H, CH2OH), 4.16 (d of q, J = J '= 6.2Hz, 1H, H-1'), 3.98 (dvont, J = 9, 1Hz, J '= 2.6Hz, IH , H-5), 3.75-3.20 (m, 3H), 3.20-2.65 (m, 2H), 1.22 (d, J = 6.4 Hz, 3H, CHMe)

UV (H20) Xmax: 294 (e 7614), 266 (e 6936) nm tv4 = (pH7.4; 36.8 ° C) 14.0h.

Example 4

Preparation of (5R, 6S) -3- [2- (4-hydroxymethylpyridinio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-azabicyclo [3.2.0] hept-2-en- 2-carboxylate oh

A,

-.3 ^

ch2oh

A.4-hydroxymethyl-l- (2-mercaptoethyl) pyridinium trifluoromethanesulfonate ch2 (m + CFjsojh + / \

hsch2ch2

+ H§) -CH2

40 min at 0 ° C, this solution is added to a solution of 4-hydroxymethyl-l- (2-mercaptoethyl) pyridinium trifluoromethanesulfonate (0.447 g, 1.4 mmol) in 1 ml acetonitrile, followed by diisopropylethylamine ( 0.191 ml, 1.1 mmol), whereby a reddish-black, rubber-like product separates from the reaction mixture, the reaction mixture is filtered after 20 min at 0 ° C. and concentrated in vacuo. The residue is taken up in the smallest possible amount of acetonitrile water (1: 1) and the mixture is added to a C18 reverse phase column. It is eluted with 25% acetonitrile water and then the relevant fractions are lyophilized, the product (0.353 g,

47%) as a cream-colored solid.

IR (KBr) vmax: 3240 (br, OH), 1775 (β-lactam (CO), 1695 (-C02PNB) cm "1 15 XH-NMR (d6-acetone) ô: 9.24-7.84 (m , 8H, aromatic), 7.4-6.9 (m, ÎOH, diphenylphosphate), 5.52.5.24 (ABq, J = 14Hz, 2H, benzylic), 5.15-4.80 (m, 4H), 4.45-3.05 (m, 7H), 1.35 (d, J = 6.6 Hz, 3H, CHMe).

20 C. (5R, 6S) -3- [2- (4-hydroxymethylpyridinio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2 -en-2-carboxylate

OH

,Oh

25th

X

CFjSOJ "

Trifluoromethanesulfonic acid (1.327 ml, 0.015 mol) is added dropwise to a solution of 4-pyridine-methanol (1.635 g, 0.015 mol) in 10 ml of CH2C12 under N2 at 0 ° C. A yellow-brown oil separates quickly. An additional equivalent of 4-pyridine methanol (1.635 g, 0.015 mol) is then added to this mixture and the solvent is removed under reduced pressure to give an oil. Ethylene sulfide (0.891 ml, 0.015 mol) is added to this oil and the homogeneous mixture obtained is heated on an oil bath at about 60 ° C. for 3 hours. The reaction mixture is then taken up in 15 ml of water and the aqueous solution is washed with CH2C12 (5x5 ml). After residual organic solvent has been removed in vacuo, the aqueous layer is placed on a CI8 reverse phase column. The mixture is eluted with water and the appropriate fractions are then concentrated, giving an oil which is dried further in vacuo over P2Os, giving the product (4.64 g, 97%) as a colorless oil.

IR (film) vmax: 3455 (st, OH), 2565 (schw, SH) CM "1

^ -NMR (d6-acetone) ô: 9.07.8.18 (ABq, J = 6.8 Hz, 4H, aromatic), 5.03 (s, 2H, CH2OH), 4.96 (1, J = 6.5 Hz, 2H, N-CH2), 4.09 (br.s, IH, -OH).

Bp-nitrobenzyl- (5R, 6S) -3- [2- (4-hydroxymethylpyridino) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-azabicyclo [3.2.0] hept-2- en-2-carboxylate diphenyl phosphate

CH, OH

30th

c02pnb ^ 0) 2p0

OH

A

CHo0H

0h

A

XT>

co-pnb

0H

c02pnb

- \ 0> -ch

0

«J0) 2P0-

ÏQ-:

A mixture of p-nitrobenzyl- (5R, 6S) -3- [2- (4-hydroxy-35 methylpyridinio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [ 3.2.0] hept-2-ene-2-carboxylate diphenyl phosphate (0.348 g, 0.465 mmol) and 10% palladium-on-carbon (0.35 g) in 11 ml phosphate buffer (0.05 M, pH 7.4 ), 5 ml of THF and 10 ml of ether are hydrogenated at 2.76 bar (40 psi) for 1.25 h. The mixture 40 is filtered through a pad of Celite, the aqueous phase is washed with ether (3 ×), the pH of the aqueous solution is adjusted to 7.0 using a further pH 7.4 buffer, residual volatiles are removed in the Vacuum and place the aqueous solution on a C18 reverse phase column. It is eluted with 2% 45 acetonitrile water and then lyophilized, giving a yellow-brown solid. This material is chromatographed again (C18 reverse phase / H20), giving the desired product (0.060 g, 36%) as a light yellow solid.

50 IR (KBr) v max: 3400 (b4, OH), 1755 (β-lactam OH), 1590 (-COf) cm "1

^ -NMR (D20) ô: 8.73.7.96 (ABq, J = 6.8 Hz, 4H, aromat.), 4.93 (s, 2H, CH2OH), 4.77 (t, J = 6.0 Hz, 2H, N-CH2), 4.15 (d of q, J = J '= 6.3 Hz, IH, H-l'), 3.96 (d of t, J = 9.2 Hz , 55 J '= 2.6 Hz, 1H, H-5), 3.65-3.20 (m, 3H), 3.13-2.62 (m, 2H), 1.21 (d, J = 6.3 Hz, 3H, CHMe)

UV (H20) km, x: 295 (e 6880), 256 (e 5595), 224 (e 8111) nm% W (pH 7.4, 36.8 ° C) 14.5 h.

60

To a solution of p-nitrobenzyl- (5R, 6S) -6- [l- (R) -hydro-xyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2-carboxylate (0.348 g , 1.0 mmol) in 5 ml of dry acetonitrile are added dropwise under N2 at 0 ° C. diisopropylethyl chlorophosphate (0.191 ml, 1.1 mmol). The resulting golden yellow solution is stirred

Example 5

Preparation of 3- [2- (l- (2-methylpyridinium)) ethylthio] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en- 2-carbo-

xylate

65

coo

658 248

22

A. 1- (2-Mercaptoethyl) -2-methylpyridinium methanesulfonate

OH

A

zfx

+ MeOH

55 C, 21 h

MsO

COOPNB

(PhO) pfû "

10g Pd / C, ed

THF, ether, buffer

Ethylene sulfide is added to a suspension of 2-methyl pyridinium methanesulfonate in 2-methyl pyridine, prepared by adding methanesulfonic acid (0.65 ml, 0.010 mol) to cold 2-methyl pyridine (2.17 ml, 0.022 mol) (0.655 ml, 0.011 mol), the reaction mixture is stirred under a nitrogen atmosphere at 55 ° C. for 21 h, cooled to 23 ° C. and diluted with 5 ml of water. The aqueous solution is washed with ether (6x4 ml). Traces of organic solvents are removed by pumping. The solution is then added to a column (2.5 x 10.0 cm) of ji-Bondapack C-18, the column is eluted with water and the appropriate fractions are lyophilized to give 2.13 g (85%) of the title compound .

IR (film) vmax: 2520 (SH), 1623 (pyridinium), 1574.1512, 1485, 1412, 1195 (sulfonate), 1038 cm "1

'H NMR (DMS0-d6 + D20) δ: 2.37 (3H, s, CH3SO3-), 2.83 (3H, s, CH3 on pyridinium), 3.09 (2H, J = 6.9 Hz , CH2S), 4.71 (2H, t, J = 6.9 Hz, CH2N +), 7.93 (2H, m, Hm of pyridinium), 8.44 (1H, m, Hp of pyridinium), 8, 89 (IH, m, Ho of pyridinium)

UV (H20) Xmax: 266 (s 3550) mu.

Bp-nitrobenzyl-3- [2- (1- (2-methylpyridinium-ethylthio) -6a [1- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en- 2-carbo-xylate-diphenylphosphate oh 1) NEt (iPr) 2

A ~ ry0 2) ClP (OPh) g

0H

A

«R +

10th

eoo

3) HS

COOPNB

COOPNB

4) NEt (iPr) 2

To a solution of p-nitrobenzyl-3- [2- (l- (2-methylpyridium-nium)) ethylthio] -6cc- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo- [3.2 .0] hept-2-en-2-carboxyiat-diphenylphosphate (0.66 g, 0.90 mmol) in moist tetrahydrofuran (34 ml) are added ether 15 (34 ml), potassium phosphate monobasic sodium hydroxide buffer (0.15 M , 16.5 ml, pH 7.22) and 10% palladium-on-carbon (0.66 g) and hydrogenated the mixture obtained for 1.25 h at a pressure of 1.76 bar (40 psi) and 23 ° C. The organic layer is separated off and extracted with buffer (2x6 ml). The water-20 layers are combined, filtered through a Celite pad, washed with ether (40 ml), traces of organic solvents are removed by applying a vacuum and applied to a u-Bondapack C18 column (2.5 × 10 cm). The column is eluted with water and the appropriate fractions are lyophilized, giving 0.098 g

(31%) of the title compound as a yellowish powder.

IR (KBr) vmax: 3650-3100 (ÖH), 1755 (C = 0 of ß-lactam), 1630 (pyridinium), 1595 cm "" 1 (carboxylate)

> H NMR (D20) ô: 1.20 (3H, d, J = 6.3 Hz, CH3CHOH), 2.83 (s, CH3 on pyridinium), 2.7-3.1 (5H, H- 4, CH3 on pyridinium), 3.1-3.7 (3H, m, CH2S, H-6), 3.90 (dd, J = 9, 1 Hz, J = 2.6 Hz, H-5) , 3.1 (m, CH3CHOH), 4.78 (t, J = 6.2Hz, CH2N +), 7.8 (2H, m, Hm from pyridinium), 8.3 (1H, m, Hp from pyridinium) , 8.65 (1H, s, Ho of pyridinium)

UV (H20) Xmax: 268 (e 3950), 196 (e 8840) m | X [a] o = + 41 °

xi /,: 15.0 h (measured at a concentration of 10 "4 M in phosphate buffer pH 7.4 at 36.8 ° C).

25th

30th

35

To a cold (0 ° C) solution of p-nitrobenzyl-6a- [l- (R) -hydroxyethyl] -3,7-dioxo-l-azabicyclo [3.2.o] heptane-2-carboxylate4o (0.523 g, 1 , 50 mmol) in acetonitrile (6 ml) (stored under nitrogen), diisopropylethylamine (0.314 ml, 1.80 mmol) and then diphenyl chlorophosphate (0.373 ml,

1.80 mmol). The reaction mixture is stirred at 0 ° C. for 30 min and treated with a solution of 1- (2-mercaptoethyl) -2- 45 methylpyridinium methanesulfonate (0.530 g, 2.16 mmol) in 18 ml of acetonitrile and then with diisopropylethylamine (0.314 ml , 1.8 mmol). The reaction mixture is stirred at 0 ° C. for 1 h, diluted with cold (0 ° C.) water (26 mol) and the mixture is placed on an a-Bondapak C18 column (3.5 × 7.0 cm). The column is eluted with 25% acetonitrile-75% water and with 50% acetonitrile-50% water. After lyophilization of the appropriate fractions, 1.06 g (96%) of the title compound is obtained as a yellowish powder.

IR (KBr) vmax: 3650-3100 (OH), 1770 (C = 0 of β-lactam), 55 1695 and 1690 (C = 0 of the PNB ester), 1630 (pyridinium), 1595 (phenyl), 1518 ( N02), 1335 (N02), 890 cm "1 (NO,)

'H NMR (DMSO-d6) Ô: 1.15 (3H, d, J = 6.1 Hz, CH3CHOH), 2.87 (s, CH3 on pyridinium), 3.6-4.4 (2H, m, H-5, CH3CHOH), 4.75 (2H, m, CH, N +), 5.37 (center of ABq, J = 14Hz, CH go of PNB), 6.5-7.4 (10H, m , Phenyl), 7.70 (2H, d, J = 8.8Hz, Ho from PNB), 8.0 (2H, m, Hm from pyridinium), 8.24 (2H, d, J = 8.8 Hz , Hm from PNB), 8.50 (IH, m, Hp from pyridinium), 8.95 (1H, br.d, J = 6, 1 Hz, Ho from pyridinium)

UV (H20) Xraax: 265 (s 11990), 314 (e8020) m y .. 6s

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

Example 6

Preparation of 3- [2- (1- (4-methylpyridinium)) ethylthio] -6a- [1- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] -hept-2-ene -2-carbo-

xylate

OH

n r +

CH,

COO

50 A. l- (2-mercaptoethyl) -4-methylpyridinium methanesulfonate zf \

Ms OH

55 C, 24 h

HS

To a suspension of 4-picolinium methanesulfonate in 4-picoline, prepared by adding methanesulfonic acid (0.65 ml, 0.010 mol) to 4-picoline (2.14 ml, 0.022 mol), ethylene sulfide (0.655 ml, 0.011 mol) is added with cooling ), the reaction mixture is stirred for 4 h under a nitrogen atmosphere at 55 ° C., cools to 23 ° C. and diluted with water (5 ml) and ether (10 ml). The organic layer is separated off, the aqueous layer is washed with ether (5 × 5 ml) and placed on an | x-Bondapak C18 column (2.5 × 10 cm) after traces of ether have been removed under reduced pressure. The column is eluted with a 15% acetonitrile-85% water mixture and the appropriate fractions are lyophilized. This gives 2.66 g (100%) of a colorless syrup.

IR (film) vmax: 2500 (SH), 1640 (pyridinium), 1572.1520, 1478.1200 (sulfonate), 1040, 833 and 768 cnT1

23

658 248

^ -NMR (DMSO-d6ô: 2.31 (3H, s, CH3S03 "), 2.62 (s, CH3 on pyridinium), 2.2-2.9 (4H, SH, CH3 on pyridinium), 3, 04 (2H, m, CH2S), 4.68 (2H, t, J = 6.4Hz, CH2N +), 8.01 (2H, d, J = 6.6 Hz, Hm from pyridinium), 8.89 ( 2H, d, J = 6.6 Hz, Ho of pyridinium)

UV (H20)) ,, mx: 256 (e 4100), 221 (e 7544) mu.

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

Ä +

+ pTfiOH

HS '

pTsO ~

4-Picolon (1.17 ml, 0.012 mol) is added to a suspension of p-toluenesulfonic acid (1.72 g, 0.01 mol) in benzene (6.5 ml), and the resulting mixture is stirred under a nitrogen atmosphere for 30 min at 23 ° C, treated with ethylene sulfide (0.65 ml, 0.011 mol) and stirred at 75 ° C for 24 h. More ethylene sulfide (0.65 ml, 0.011 mol) is added, the mixture is stirred at 75 ° C. for a further 24 h, the reaction mixture is cooled to 23 ° C. and diluted with 5 ml of water and 8 ml of ether. The aqueous layer is separated off, washed with ether (3 × 8 ml), traces of organic solvents are removed in vacuo and chromatographed on [X-Bonda-pak C-18 using water as the eluent. 2.94 g (90%) of the title compound are obtained as a colorless syrup.

IR (film) vmax: 2510 (SH), 1640 (pyridinium), 1595.1582, 1475, 1475, 1200 (sulfonate), 1031.1010, 818 cm "1

1H NMR (DMSO-d6ô: 2.29 (3H, s, CH3 on pyridinium), 2.61 (s, CH3Ph), 2.4-2.8 (4H, SH, CH3Ph), 3.03 (2H , m [addition of D20 gives a t, J = 6.4 Hz, at 3.04], CH2S, 4.68 (2H, t, J = 6.4 Hz, CH2N +), 7.11.7.49 (4H, 2d, J = 7.9Hz, phenyl), 8.00 (2H, d, J = 6.5 Hz, Hm of pyridinium), 8.89 (2H, d, J = 6.5 Hz, Ho of pyridinium)

UV (H20) Xmax: 256 (e 4315), 222 (s 17045) mu

C. p-nitrobenzyl-3- [2- (l- (4-methylpyridinium)) ethylthio] -6 <x- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2-car boxylate diphenyl phosphate

1) NEt (iPr)

CH2N +), m5.17 (d, J = 4.9 Hz, OH), 5.37 (center of ABq, J = 14.1 Hz, CH2 of PNB), 6.7-7.4 (10H, m, phenyl ), 7.69 (2H, d, J = 8.8 Hz, Ho from PNB), 8.00 (2H, d, J = 6, m5 Hz, Hm from pyridinium), 8.23 (2H, d, J = 8.8 Hz, Hm from PNB), 8.92 (2H, d, 5 J = 6.5 Hz, Ho from pyridinium)

UV (H20) Xmax: 262 (e 10835), 311 (e 9670) mu Analysis: for C36H36N3Oi0SP-r, 5H2O calculated: C 56.84%, H 5.17%, N 5.52%, S 4.21 % found: C 56.89%, H 5.13%, N 5.19%, S 4.41%

io

D.3- [2- (1- (4-methylpyridinium)) ethylthio] -6a- [1- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2- en-2-carboxylate

COOPNB

(PhO) -PO COOPNB 2 "

0

OH

15

20th

A

M *

COOPNB

(Ph0)? £ 0 *

10 # Pd / C,

THF, ether, buffer

OH

To a cold (0 ° C) solution of p-nitrobenzyl-6a- [l- (R) -hydroxyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2-carboxylate (0.522 g, 1 , 5 mmol) in acetonitrile (6 ml) (stored under nitrogen), diisopropylethylamine (0.314 ml, 1.8 mmol) and then diphenyl chlorophosphate (0.373 ml, 1.9 mmol) are added, the reaction mixture is stirred for 45 min and treated dropwise with it a solution of 1- (2-mercaptoethyl) -4-methyl pyridium methanesulfonate (0.539 g, 2.16 mmol) in acetonitrile (1.8 ml) and then with diisopropylethylamine (0.314 ml, 1.8 mmol). The reaction mixture is stirred for 1 h at 0 ° C., diluted with cold (0 ° C.) water (24 ml) and applied to an (x-Bondapak-C18 column (2.5 × 8.5 cm). The column is eluted first with a 25% acetonitrile-75% water mixture (100 ml), then with a 50% acetonitrile-50% water mixture (100 ml) and after lyophilization of the appropriate fractions 0.91 g (83%) of Title compound as a yellowish powder. __

IR (KBr) vmax: 3700-2800 (OH), 1770 (C = O of β-lactam), 1700 (C = 0 of the PNB ester), 1640 (pyridinium), 1595 (phenyl), 1520 (N02), 1340 (N02), 890 cm "1 (N02)

MMR (DMSO-d6) Ô: 1.16 (3H, d, J = 6.2Hz, CH3CHOH), 2.61 (s, CH3 on pyridinium), 3.1-3.7 (3H, m, H- 6, CH2S), 3.7-4.4 (2H, m, H-5, CH3CHOH), 4.79 (2h, br.t, J = 6.3 Hz,

eoo

To a solution of p-nitrobenzyl-3- [2- (1- (4-methylpyridium-nium)) -ethylthio] -6a- [1- (R) -hydroxyethyl] -7-oxo-l-azabicyclo-25 [ 3.2.0] hept-2-en-2-carboxylate-diphenylphosphate (0.587g, 0.80 mmol) in moist tetrahydrofuran (30 ml) are added ether (30 ml), potassium phosphate-monobasic sodium hydroxide buffer (0.15 M , 14.7 ml, pH 7.22) and 10% palladium-on-carbon (0.59 g), the resulting mixture is hydrogenated for 1.25 h at 23 ° C. under 30 a pressure of 1.76 bar (40 psi) , separates the organic layer and extracted with the buffer (2x6 ml). The aqueous extracts are combined, filtered through a pad of Celite, washed with ether (3 x 20 ml), traces of organic solvents are removed by pumping and placed on an [x-Bondapak C-18 column 35 (2.5 x 10 cm ). Elution of the column with water and lyophilization of the appropriate fractions gives 0.135 g (49%) of the title compound as a yellowish powder.

IR (KBr) vmax: 3700-3000 (OH), 1770 (C = 0 of ß-lactam), 1642 (pyridinium), 1592 cm-1 (carboxylate)

40 'H NMR (D20) Ô: 1.19 (3H, t, J = 6.3 Hz, CH3CHOH), 2.59 (3H, s, CH3 on pyridinium), 2.84 (d, J = 9 , l Hz, H-4), 2.90 (d, J = 9, l Hz, H-4), 3.0-3.6 (3H, m, CH2S, H-6), 3.86 ( dd, J = 9, 1 Hz, J = 2.6 Hz, H-5), 4.12 (m, CH3CHOH), 4.5-4.9 (CH2N +, masked by HOD), 7.80 (2H , d, J = 6.6 Hz, Hm of pyridi-45 nium), 8.58 (2H, d, J = 6.6 Hz, Ho of pyridinium)

UV (H20)> .max: 256 (e 5510), 262 (e 5360), 296 (e 7050) nm [a] o = + 20.89 ° (C 0.48, H? 0)

12.8 h (measured at a concentration of 10 "4 M in a phosphate buffer pH 7.4 at 36.8 ° C).

50

Example 7

Preparation of (5R) -3- [2- (4-methylthiopyridinio) ethylthio] - (6S) - [(IR) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en- 2-carboxylate

55

60

CO «"

65

§) -SH

A. 4-methylthiopyridine +

1) MeJ / EtOH tfA \ SHe

2) NaOH

4-Mercaptopyridine (5.55 g, 50.0 mmol; Aldrich) is dissolved in 50 ml of boiling absolute ethanol. The insoluble material is filtered off through Celite. The filtrate is heated to

658 248

24th

Dissolve again and, when it has cooled to about 50 ° C., methyl iodide (3.17 ml, 51.0 mmol; Aldrich) is added all at once. The mixture is cooled to crystallize out. The solid is filtered off, giving 6.77 g (26.7 mmol; yield 53.5%) of the title compound as the hydroiodide.

'H NMR (DzO) ô: 2.70 (3H, s, -SCH3) and 7.65-7.77-8.35-8.48 ppm (4H, A2B2 type, aromatic Hs) IR ( Nujol) vmax: 1615.1585 (aromat.) And 780 cm-1 UV (H, 0) Xmax: 227 (e 2.02xl04) and 298 nm_ (el, 64xl04). The hydroiodide (6.33 g, 25.0 mmol) is dissolved in 40 ml of water, insoluble material is removed, washed with 10 ml of water, added to the filtrate at 0 to 5 ° NaOH pellets (5 g) and extracted with EtzO (3 x25 ml), the aqueous layer being saturated with NaCl, the combined organic extracts washed with brine (2x), dried (MgS04) and concentrated to give 2.92 g (23.4 mmol, overall yield 50%) of the title compound received as oil.

'H NMR (CDC13) Ô: 2.48 (3H, s, -SCH3) and 7.03-7.13-8.38-8.48 ppm (4H, A2B2 type, aromatic Hs) IR ( Film) vmax: 1580 and 800 cm "" 1

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

B.4-Methylthio-N- (2-mercaptoethyl) pyridinium methylsulfonate

V

CHJSOJH

(475 mg, 1.36 mmol) and diisopropylethylamine (0.24 ml, 1.4 mmol) in 5 ml of CH3CN are added under a nitrogen atmosphere at 0 to 5 ° C. diphenyl chlorophosphate (0.29 ml, 1.41 mmol) the mixture for 30 min at 0 to 5 ° C. gives an oily suspension of 4-methylthio-N- (2-mercaptoethyl) pyridinium methanesulfonate (678 mg, 1.45 mmol; 60% purity) to this mixture 1.5 ml of CH3CN and then diisopropylethylamine (0.24 ml, 1.4 mmol) and the mixture is stirred for 1 h at 0 to 5 ° C. Immediately after adding the base, a yellow precipitate forms. The precipitate is filtered off and washed with 3 ml of cold CH3CN to give 413 mg of a yellowish solid. This is triturated with 10% MeOH in water (5 ml), giving 341 mg (0.618 mmol, yield 45.4%) of the title compound as white crystals; Mp 118 to 120 ° C.

15 'H NMR (DMSO-de, CFT-20) Ô: 1.16 (3H, d, J = 6, 1 Hz, l'-CH3), 2.72 (3H, s, -SCH3), 3 , 1-3.7 (5H, m), 3.7-4.3 (2H, m), 4.71 (2H, t, J = 6.3 Hz, -CH2N +), 5.15 (1H, d, J = 4.9 Hz, OH), 5.20-5.35-5.40-5.55 (2H, ABq, C02CH2-Ar), 7.70 (2H, d, J = 8.8 Hz, nitrophenyl-Hs), 7.97 (2H, d, J = 7.0Hz, pyridinio-20 Hs), 8.25 (2H, d, J = 8.8 Hz, nitrophenyl-Hs) and 8.76ppm ( 2H, d, J = 7.1 Hz, pyridinio-Hs)

IR (Nujol) vmM: 3250 (OH), 1775 (ß-lactam), 1700 (ester) and 1625 cm "1 (pyridinio)

UV (abs. EtOH) 308 nm (e 4.47x104) 25 [a] g = + 24.8 ° (c 0.5, MeOH)

Analysis: for C24H26N3Of) SiCI • H20 calculated: C 50.56%, H 4.95%, N 7.37%

found: C 5.63%, H 4.72%, N 67.89%

50-60 '

30th

4-Methylthiopyridine (2.75 g, 22.0 mmol) is slowly added to methanesulfonic acid + (0.65 ml, 10.5 mmol) while cooling in an ice bath. Ethylene sulfide "1" (0.66 ml, 11.0 mmol; Aldrich) is added to this solid and the mixture is heated at 50 to 60 ° C. for 21 hours. As the implementation progresses, the solid goes into solution. After cooling, the reaction mixture is dissolved in 5 ml of water, washed with Et20 (5x4 ml), the fluffy, aqueous layer is filtered through Celite and the filtrate is purified by reverse-phase silica gel column chromatography (C18 micro Bondapak 10 g), whereby eluted with water. Fractions of 10 ml were collected in each case. Fractions 2 and 3 are combined and cleaned again with the reverse phase column. Fraction 2 gives 1.258 g (4.48 mmol; yield 42.6%) of the title compound as a viscous oil.

■ H NMR (DMSO-d6, CFT-20) ô: 2.32 (3H, s, MeSOf), 2.72 (3H, s, -SMe), 2.68 (IH, m, SH), 2 , 9-3.2 (3H, m, -CH2S-), 4.59 (2H, t, J = 6.4 Hz, -CH2N +), 7.97 (2H, d, J = 7.2 Hz, aromatic Hs) and 8.72 ppm (2H, d, J = 7.2 Hz, aromatic Hs)

IR (neat) vmax: 1630, 1200 (br., -S03-), 785 and 770 cm "1 + These reagents were distilled before use.

C. (5R) -p-nitrobenzyl-3- [2- (4-methylthiopyridino) ethylthio] - (6S) - [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2-carboxylate chloride

D. (5R) -3- [2-methylthiopyridinio) ethylthio] - (6S) - [(IR) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2 carboxylate

A

35

40

SMe

Cl

H2 / Pd-C

PH 7.4 buffer

0H

CO «"

• SMe

45

OH

A

£]>

1) ClPO «tf) -

HbO "

■ SMe

-SMe

To a solution of (5R) -p-nitrobenzyl-3,7-dioxo (6S) - [(IR) -hydroxyethyl] -l-azabicyclo [3.2.0] heptane (2R) carboxylate

(5R) -p-nitrobenzyl-3 - [- methylthiopyridinio-ethylthio] - (6S) - [l (lR) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2- car-boxylate chloride (380 mg, 0.688 mmol) is dissolved in 31.5 ml THF and a pH 7.40 phosphate buffer (31.5 ml, 0.05M; Fisher), diluted with Et20 (31.5 ml), mix this solution with 10% Pd-C (380 mg; Engelhard) and hydrogenate for 1 h at room temperature with a Parr shaker at 2.4 bar (35 psi). The aqueous layer is filtered through Celite to remove the catalyst and the Celite pad is washed 50 with water (2x5 ml). The filtrate and the wash water are combined and washed with Et20 (2x30 ml). The aqueous layer is freed from organic solvents under reduced pressure and purified by reverse phase column chromatography (C18 microbondapak, 13 g; Waters 55 Associates), eluting with water. Fractions with UV absorption at 307 nm are collected (approx. 11) and lyophilized, giving 127 mg (0.334 mmol, yield 48.5%) of the title compound as a yellowish powder.

* H-NMR (DiO, CFT-20) Ô: 1.20 (3H, d, J = 6.4Hz, l'-CH3), 60 2.64 (3H, s, -SCH3), 2.81 ( 2H, m, -SCHr), 3.19 (1H, dd, Jw. = 6, 1 Hz, JM = 2.6 Hz, 6-H), 3.32 (2H, dd, J = 11 Hz, J = 5.5 Hz, 4-Hs), 3.92 (1H, dt, J = 9.2Hz, J5 ^ = 2.6 Hz, 5-H), 4.1 (1H, m, l'-H ), 4.61 (2H, t, J = 5.9 Hz, -CH2N +), 7.70 (2H, d, J = 7.1 Hz, aromatic Hs) and 8.40 ppm (2H, d, J = 7.1 Hz, aromatic Hs) 65 IR (KBr, pill) vmas: 3400 (OH), 1750 (ß-lactam), 1630 (pyridinium) and 1590 cm "" 1 (carboxylate)

UV (H-.O) lmax: 231 (e 9800) and 307 nm (e 25000) [ct] 2D3 = + 3.14 ° (c, 0.5; H20)

25th

658 248

Example 8 c. 3- [2-methoxy-l-pyridinium) ethylthio] -6a [l '- (R) -hydroxy-

Preparation of 3- [2- (3-methoxy-l-pyridinium) ethylthio] -6a-ethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-ene-2-carboxylate [l '- (R ) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2-car-

boxylate 0H C1 0He

OH

OMe r

C0o

10th

GH

OMe

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

MeO

Meo

-Q • a-

MSO

HsOH

To pre-cooled (5 ° C) 3-methoxypyridine (698 mg,

6.4 mmol) of methanesulfonic acid (0.216 ml,

3.05 mmol) and ethylene sulfide (0.19 ml, 3.2 mmol). The mixture is then heated to 60 ° C. for 18 h, cooled to 20 ° C., diluted with water (10 ml) and washed with ether (3 × 10 ml). The aqueous phase is treated under high vacuum for 15 min and placed on a Ci8 reverse phase column. The title compound is eluted with water. The appropriate fractions are combined and concentrated under high vacuum, giving the desired thiol (61.6 mg, yield 76.3%).

IR (CH2a2) vŒM: 2550 (black, SH) and 1620.1600.1585 cm "1 (m, aromat.)

: H-NMR (DMSO-d6) Ô: 8.90-7.90 (4H, m, aromatic CH), 4.72 (2H, t, J = 6.6 Hz, CH2N +), 4.01 ( CH, s, OCH3), 3.5-3.0 (m, hidden-CH2S), 2.66 (1H, dd, J = 9.5 Hz, J = 7.5 Hz, SH) and 2.31 ppm (3H, s, CH3S03-).

B. p-nitrobenzyl-3- [2- (3-methoxy-l-pyridinium chloride) -ethyl-thio] -6a- [r- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2-carboxylate

1) Etu «) 2

2) ClP0 {0rf) 2

C02

A solution of p-nitrobenzyl-3- [2- (3-methoxy-l-pyridi-15 nium chloride) ethylthio] -6a- [l '- (R) -hydroxyethyl] -7-oxo-l-aza-bicyclo [3.2.0] hept-2-en-2-carboxylate (600mg, 1.12 mmol) in THF (25 ml), ether (25 ml) and pH 7.4 phosphate buffer (0.1 M, 25 ml) hydrogenated one hour in a Parr shaker at 2.76 bar (40 psi) over 10% Pd / C. The mixture is diluted with ether 20 and the aqueous phase is filtered through a No. 52 hardened filter paper. The aqueous layer is washed with ether (2 x 20 ml), a vacuum is applied and applied to a silica gel reversed phase column. The title compound is eluted with water containing 2% and 5% acetonitrile. The appropriate fractions are combined and lyophilized to give a yellow solid which is purified by HPLC to give the penem carboxylate (150 mg, 38%).

IR (Nujol) vmax: 1750 (st, ß-lactam C = 0) and 1580 cm "1 (st, carboxylate)

30 ^ -NMR (D20) ô: 8.55-8.30, (2H, m, H-2, H-6 aromat.), 8.17-7.75 (2H, m, H-3, H -4 aromat.), 4.77 (2H, t, J = 5.9 Hz, CH2N +), 4.10 (1H, part of 5 lines, J = 6.3 Hz, H-l '), 3, 97 (3H, s, OCH3), 3.85, 3.82 (2 lines, part of dt, J = 2.6 Hz, part of H-5), 3.42 (2H, t, J = 5.9 Hz, CH2-S), 3.25 (1H, dd, J = 6, 1 Hz, J = 2.6 35 Hz, H-6), 2.99-2.60 (2H, 6 lines, part of H- 4) and 1.20 ppm (3H, d, J = 6.4 Hz, CH3)

UV (H20, c 0.05) 290 (e 10517), 223 (e 6643) tv ,: (0.1 M pH 7.4 phosphate buffer, 37 ° C) = 20 h.

40

OMe

CMe

Example 9

Preparation of (5R, 6S) -3- [2- (3-Methylthiopyridinio) -ethyl-thio] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept- 2-en-2-carboxylate

45

0 «

A

A cold (0 ° C) solution of p-nitrobenzyl-6a- [l '- (R) -hydroxyethy] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2-carboxylate (1.04 g , 3 mmol) in 12 ml acetonitrile is treated dropwise with diisopropylethylamine (0.63 ml, 3.6 mmol) and diphenyl chlorophosphate (0.75 ml, 3.6 mmol) and stirred for 30 min at 0 ° C. The enol phosphate obtained is treated with 1- (2-mercapto-ethyl) -3-methoxypyridinium methanesulfonate (1.14 g,

4.30 mmol) in 7 ml CH3CN, 0.63 ml (4.30 mmol) diisopropylethylamine, stirred for 30 min and cooled to -10 ° C for 30 min. The precipitated solid is filtered off from the mixture, washed with cold acetonitrile (2 ml) and dried to give the title compound (1.32 g, yield 82%).

IR (Nujol) vmax: 3320 (m, OH), 1780.1765 (st., Β-lactam C = 0), 1700.1695 (m, ester C = 0) and 1520 CM "1 (st, N02)

! H NMR (DMSO-d6) Ô: 9.01 (1H, bs, H-3 aromat.), 8.75 (1H, bd, J = 5.4Hz, H-6 aromat.) 8.35- 7.95 (4H, m, H-aromat.), 7.70 (2H, d, J = 7.7 Hz, H-aromat.), 5.37 (2H, center of ABq, J = 13 Hz, CH2PNB), 5.17 (1H, d, J = 4.9 Hz, OH), 4.87 (2H, t, J = 6.3 Hz, CH, -N +), 4.35-3.75 ( 2H, m, H-5 and H-l '), 4.00 (3H, s, OCH3), 3.56 "(part of t, J = 6.3 Hz, CH2S), 3.5-3, 20 (3H, m, H-6, H-3) and 1.16 ppm (3H, d, J = 6, 1 Hz, CH3CHO).

CO,

50

55

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

SMe

SMe

HCl

À

Cl

To a solution of 3-methylthiopyridine (2.00 g, 0.016 mol) (prepared by the method of JA Zoltewiez and C. Nisi, J. Org. Chem. 34,765,1969) in 10 ml of ether is added 15 ml of IN HCl, shake the mixture thoroughly, separate the aqueous phase 60, wash with 10 ml ether and then concentrate. The remaining hydrochloride is then dried in vacuo (P2Os), giving a white solid. 3-Methylthiopyridine (1.88 g, 0.015 mol) and ethylene sulfide (0.89 ml, 0.015 mol) are added to this hydrochloride solid and the 65 mixture obtained (oil bath) is heated at 55 to 65 ° C. under N2 for 15 h. A slightly cloudy oil is obtained, which is taken up in 125 ml of water and washed with CH2C12. The aqueous solution is concentrated to about 25 ml, then a few drops of acetonitrile are added to the mixture

658 248

26

To make the solution homogeneous, put the aqueous solution obtained on a C18 reverse phase column, elute with water and then concentrate the relevant fractions, giving the product as a pale yellow, viscous oil (2.66 g, 80%). IR (film) vmax: 2410 (br, -SH) cm-1 'H-NMR (d6-DMSO + D2ö) δ: 8.88-7.88 (m, 4H, aromat.), 4.70 (t , J = 6.5 Hz, 2H, N-CH- »), 3.08 (oblique t, J = 6.5 Hz, 2H, S-CH2), 2.64 (s, 3H, S-Me)

B. p-Nitrobenzyl- (5R, 6S) -3- [2- (3-methylthiopyridinio) ethylthio] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2. 0] hept-2-en-2-carboxylate chloride

OH

A solution of p-nitrobenzyl- (5R, 6S) -6- [l- (R) -hydroxy-ethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2-carboxylate (0.522 g, 1.50 mmol) in 7 ml of dry acetonitrile is cooled to 0 ° C., then diisopropylethylamine (0.287 ml, 1.65 mmol) is added dropwise, and dropwise diphenyl chlorophosphate (0.342 ml, 1.65 mmol) is added to the yellow-brown solution obtained. and hold the reaction mixture at 0 ° C for 30 min. Diisopropylethylamine (0.313 ml, 1.80 mmol) is then added, followed by a solution of 3-methylthio-l- (2-mercaptoethyl) pyridium chloride (0.398 g, 1.80 mmol) in 0.70 ml dry DMF . About 1 min after the addition has ended, a precipitate separates from the reaction mixture. Upon further cooling to -10 ° C for 10 min, a solid, orange mass is obtained. This solid is then triturated with acetonitrile and the residue is filtered off. The residue is washed with acetonitrile, then acetone and finally dried in vacuo to give the product (0.455 g, 55%) as an off-white solid. The combined filtrate is concentrated to give a yellow oil which is taken up in as little acetonitrile as possible and cooled to 0 ° C. for 30 minutes. This mixture is filtered and a further 0.139 g of the product is obtained as a light yellow solid. The overall yield is 0.594 g (72%).

IR (KBr) v max: 3345 (br, -OH), 1770 (β-lactam CO), 1680 (-CO, PNB) cm "1

'H NMR (d6-DMSO) Ô: 8.98-7.96 (m, 4H, pyridinium aromat.), 8.20-7.65 (ABq, J = 7.0Hz, 4H, PNB aromat.) 5.53-4.80 (m, 4H), 4.3-3.7 (m, 2H), 3.6-3.25 (m, 6H), 2.66 (s, 3H, S-Me ), 1.16 (d, J = 6.0 Hz, 3H, CHMe).

C. (5R, 6S) -3- [2- (3-Methylthiopyridinio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept- 2-en-2-carboxylate

OH

A

l-azabicyclo [3.2.0] -hept-2-en-2-carboxylate chloride (0.551 g, 1.0 mmol) and 10% palladium-on-carbon (0.55 g) in 25 ml phosphate buffer (0, 05 M, pH 7.4), 5 ml of THF and 25 ml of ether are added. The mixture is hydrogenated (Parr) at 2.76 bar (40 psi) for 1 h. 5 The reaction mixture is then filtered through Celite and the filter cake is washed with water and ether. The aqueous phase is separated off, washed with additional ether (3 ×), residual organic solvents are removed in vacuo and the aqueous solution is cooled to 0 ° C. and the pH is adjusted to 7.0 with 10 saturated aqueous NaHC03. This solution is immediately placed on a C18 reverse phase column, eluted with water and then lyophilized the appropriate fractions, whereby 0.25 g of a light yellow solid is obtained. This material is again purified by reverse phase 15 HPLC to give the product (0.210 g, 55%) as a light yellow solid.

IR (KBr) v max: 3400 (br, -OH), 1755 (β-lactam CO), 1590 (-C02-) cm "1

1H-NMR (D, 0) ô: 8.60-7.76 (m, 4H, aromatic), 4.76 (t, J = 5.8 20 Hz, 2H, N-CH2), 4.13 (dvonq, J = J '= 6.3Hz, 1H, H-1'), 3.95 (d vont, J = 9.0Hz, J '= 2.8 Hz, 1H, H-5), 3, 45-2.75 (m, 5H), 2.59 (s, 3H, S-Me), 1.20 (d, J = 6.4 Hz, 3H, CHMe)

UV (H20) /.max: 296 (s 8509), 273 (e 13005), 231 (e 11576) nm ii / ,: (pH 7.4, 36.8 ° C) = 20 h.

25th

30th

Example 10

Preparation of 3- [2- (l- (2,6-dimethylpyridinium)) ethylthio] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2- en-2-carboxylate

35

40

COO

A. l- (2-Mercaptoethyl) -2,6-dimethylpyridinium methansulfonate

+ n

••• .'1a OH

100 ° C, 42 h

- »ms

MsO

45

SMa

Cl

OH

A

CO,

To a mixture of p-nitrobenzyl- (5R, 6S) -3- [2- (3-methylthiopyridinio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo

A mixture of 2,6-dimethylpyridine (19.2 ml, 0.165 mol) and methanesulfonic acid (3.27 ml, 0.050 mol) is stirred for 15 minutes, treated with ethylene sulfide (4.17 ml, 0.070 mol) and stirred in for 42 hours Nitrogen atmosphere at 100 ° C. After cooling to 25 ° C., the reaction mixture is diluted with ether (45 ml) and water (30 ml), the two layers are separated and the 50 organic layer is extracted with water (2x5 ml). The aqueous layers are combined, filtered through a Celite pad, washed with ether (2 x 15 ml), traces of organic solvents are removed in vacuo and poured onto an | x-Bondapak C-18 column (3.0 x 12 cm) . Elute with a 3% acetonitrile-97% water-55 mixture and lyophilize the appropriate fractions to give 2.5 g of the impure title compound as a syrup. This syrup is again purified by HPLC (jx-Bondapak C-18), whereby 0.90 g (7%) of the title compound is obtained.

IR (film) vmax: 2520 (SH), 1640 and 1625 (pyridinium), 1585, 60 1490, 1200 cm "1 (sulfonate)

'H NMR (DMSO-d6 + D2) Ô: 2.36 (3H, s, CH3SO3-), 4.62 (2H, m, CH2N +), 7.74 (2H, m, Hm from pyridinium), 8 , 24 (1H, m, Hp of pyridinium)

UV (H20)> .max: 272 (e 4080) m | i

65

Bp-nitrobenzyl-3- [2- (l- (2,6-dimethylpyridinium)) ethylthio] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept- 2-en-2-carboxylate diphenyl phosphate

27th

658 248

OH

^ Xì>

1) NEt (lPr) 2

0

2) ClP (OPh) p

3)

hs-

»K

[80

4) NEt (iPr) 2

COOPNB

COOPNB

(PhO)

0 «-

> P0

IR (KBr) vmax: 3700-2800 (OH), 1750 (C = 0 of β-lactam), 1620 (pyridinium), 1585 cm-1 (carboxylate)

* H-NMR (D20) ò: 1.23 (3H, d, J = 6.4Hz, CH3CHOH), 2.5-3.5 (UH, H-4, H-6, CH2S, 2CH3 on pyridinium) , 3.8-4.4 s (2H, CH3CHOH, H-5), 4.5-4.9 (CH2N +), HOD), 7.64 and 7.74 (2H, A part of the A2B system, Hm of pyridinium), 8,07,8,16, 8,18 and 8,27 (IH, B part of the A2B system, Hp of pyridinium) UV (H20) À.max: 277 (e 9733), 300 ( e 8271) m | x [a] o = + 50.7 ° (C 0.48, H20)

io analysis: for C18H22N204S-1,5 H20

calculated: C 55.51%, H 6.47%, N 7.19%

found: C 55.14%, H 6.23%, N 6.46%

To a cold (0 ° C) solution of p-nitrobenzyl-6a- [l- (R) -hydroxyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2-carboxylate (0.658 g, 1 , 89 mmol) in 6 ml acetonitrile (stored under a nitrogen atmosphere), diisopropylethylamine (0.394 ml, 2.26 mmol) and diphenyl chlorophosphate (0.468 ml, 2.26 mmol) are added, the reaction mixture is stirred for 30 min and treated with a solution of 1- (2-Mercaptoethyl) -2,6-dimethylpyridinium methanesulfonate (0.720 g, 2.73 mmol) in 3 ml acetonitrile and then with diisopropylethylamine (0.394 ml, 2.26 mmol). The reaction mixture is stirred at 0 ° C. for 2 h, diluted with cold (0 ° C.) water (27 ml) and poured onto a u-Bondapak C-18 column (2.5 × 9.0 cm). Elute with an acetonitrile-water mixture and lyophilize the appropriate fractions to give 0.92 g (65%) of the title compound.

IR (KBr) vmax: 3700-3000 (OH), 1765 (C = 0 of ß-lactam), 1690 (C = 0 of the PNB ester), 1620 (pyridinium), 1590 (phenyl), 1517 (N02), 1330 (N02), 880 cm "1 (N02)

^ -NMR (DMSO-d6) ö: 1.15 (3H, d, J = 6.2Hz, CH3CHOH), 2.7-3.7 (11H, CH2S, 2-CH3 on pyridinium, H-4, H -6), 3.7-4.4 (2H, CH3CHOH, H-5), 4.7 (2H, m, CH2N +), 5.14 (1H, d, J = 4.5 Hz, OH), 5.37 (center of ABq, J = 13.2Hz, CH2 of PNB), 6.7-7.5 (10H, m, phenyl), 7.5-8.7 (7H, pyridinium, H of PNB) UV ( H20) Xmax: 274 (el4150), 319 (e9445) m [x

15

20th

Example 11

Preparation of (5R, 6S) -3- [2- (2-methylthio-3-methylimide-azolio) -ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo- [3.2 .0] hept-2-en-2-carboxylate

25 A. 2-Methylthio-3-methyl-l- (2-mercaptoethyl) imidazolium tri-fluoromethanesulfonate

30th

* A

■ A.

w

I.

Hey

CF3S03

Trifluoromethanesulfonic acid (1.38 ml, 0.015 mol) is added dropwise to 2-methylthio-l-methylimidazole (4.0 g, 0.03 mol) [prepared as described by A. Wohl and W. Marckwald in Chem. Ber.

35 22.1353 (1889), described] at 0 ° C. under N2, then adds ethylene sulfide (0.9 ml, 0.015 mol), heats the mixture under N2 at 55 ° C. for 24 h, triturates the reaction mixture with ether (3 X) and takes up the residue in acetone. Then

"R" ri is filtered and evaporated. The product (4.2 g,

^ [{i {/ n 1; yPuyn <wÄu Yl-thio] -6a- [l- (R) -hy- 40 82%) as a semi-crystalline solid, which can be used without further droxyethyl] -7-oxo-l-azabicyclo [3.2.0] 2-en-2-carboxylate

Cleaning used.

IR (film) vmax: 2550 (black sh.) Cm "" 1 ^ -NMR (d6-acetone) Ô: 7.97 (s, 2H), 4.66 (t, J = 7 Hz, 2H, methylene ), 4.17 (s, 3H, N-Me), 3.20 (d of t, J = 7Hz, J '= 9 Hz, 45 2H, methylene), 2.72 (s, 3H, S-Me ), 2.20 (t, J = 9 Hz, 1H, -SH).

B. p-nitrobenzyl- (5R, 6S) -3- [2- (2-methylthio-3-methylimide-azolio) -ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l- azabicyclo- [3.2.0] hept-2-ene-2-carboxylate diphenyl phosphate

50

eoo

To a solution of p-nitrobenzyl-3- [2- (l- (2,6-dimethylpyridinium)) ethylthio] -6a- [l- (R) -hydroxyethyl] -7-oxo-l-azabicy- clo [3.2.0] hept-2-en-2-carboxylate-diphenylphosphate (0.80 g, 1.07 mmol) in moist tetrahydrofuran (42 ml) are added ether (42 ml), potassium phosphate-monobasic sodium hydroxide puf- fer (0.15 M, pH 7.22.21 ml) and 10% palladium-on-carbon (0.80 g), the mixture obtained was hydrogenated for 1 h at 23 ° C. and a pressure of 2.76 bar (40 psi) and filtered through a Celite cushion. The two layers are separated and the organic layer is extracted with the buffer (3x8 ml). The aqueous phases are combined, washed with 50 ml of ether, traces of organic solvents are removed in vacuo and applied to an [x-Bondapak C-18 column (3.0 = 10.2 cm). The column is eluted with a 5% acetonitrile-95% water mixture and the appropriate fractions are lyophilized to give 0.246 g (63%) of the title compound as a yellowish powder.

55

To a solution of p-nitrobenzyl- (5R, 6S) -6- [l- (R) -hydro-xyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptan-2-en-2- Carboxylate (1.40 g, 4.0 mmol) in 50 ml dry acetonitrile is added dropwise under N2 at 0 ° C diisopropylethylamine (0.76 ml, so 4.4 mmol) and then diphenyl chlorophosphate (0.91 ml, 4, 1 mmol), the reaction mixture is stirred for 1 h at room temperature, gives diisopropylethylamine (0.76 ml, 4.4 mmol) and then a solution of 2-methylthio-3-methyl-l- (2-mercaptoethyl) imidazolium trifhioromethanesulfonate ( 2.0 g, 5.9 mmol) in 5 ml of 65 acetonitrile dropwise, the reaction mixture is kept at room temperature for 1.5 h and then concentrated in vacuo to give a gummy product. This gummy product is taken up in water and put on a C18

658 248

28

Reverse phase column. It is eluted with water, then 20% acetonitrile water and finally 30% acetonitrile water. The appropriate fractions are then lyophilized and the product (0.90 g, 30%) is obtained as a light yellow solid. IR (KBr) vmax: 3380 (br, OH), 1770 (β-lactam CO) cm-1] H NMR (d6-acetone) Ô: 8.35 (brs, 1H, 8.24.7.78 ( ABq, J = 8.8 Hz, 4H, aromat.), 7.89 (brs, 1H), 7.25-6.91 (m, 10H, diphenylphosphate), 5.50.5.25 (ABq, J = 12 Hz, 2H, benzyl.), 4.75-4.27 (m, 3H), 4.03 (s, 3H, N-Me), 4.15-2.75 (m, 8H), 2 , 59 (s, 3H, S-Me), 1.22 (d, J = 6.2 Hz, 3H, -CHMe).

C. (5R, 6S) -3- [2- (2-methylthio-3-methylimidazolio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2-carboxylate

SMe w

N-Me

(<j> 0) £ p0 '

the hydrochloride is obtained as an oil, 3-aminopyridine (1.32 g, 0.015 mol) and ethylene sulfide (0.89 ml, 0.015 mol) are added to this oil and the mixture obtained (oil bath) is heated to 60 to 75 under N2 for 2 h ° C. Another equivalent of ethylene sulfide (0.89 ml, 5 0.015 mol) is added and the mixture is further heated at 55 to 65 ° C. for 65 h. The reaction mixture is washed with CH2CI2 and then taken up in 25 ml of water. The aqueous solution is placed on a Clg reverse phase column, which is eluted with water. After evaporation of the relevant fractions, the product (1.26 g, 44%) is obtained as a colorless, viscous oil.

IR (film) vmax: 3180 (NH2) cm "1] H-NMR (d6-DMSO) Ô: 8.19-7.59 (m, 4H, aromat.), 4.59 (t, J = 6, 2 Hz, 2H, N-CH2), 3.5 (brs. 2H, -NH2), 3.20-2.77 (m, 3H).

15 B. p-Nitrobenzyl- (5R, 6S) -3- [2- (3-aminopyridinio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0 ] hept-2-en-2-car-boxylate-diphenylphosphate

8H "

OH

/ "(- r \

K-Me

20th

25th

and Y

To a solution of p-nitrobenzyl- (5R, 6S) -3- [2- (2-methylthio-3-methylimidazolio) ethylthio] -6- [i- (R) -hydroxyethyl] -7-oxo l-azabicyclo [3.2.0] hept-2-en-2-carboxylate-diphenylphosphate (1.20 g, 1.56 mmol) in a mixture of 70 ml, THF, 70 ml ether and 31 ml phosphate buffer (0 , 05 M, pH 7.4), 1.2 g of 10% palladium-on-carbon are added, the mixture (Parr) is hydrogenated for 55 min at a pressure of 2.41 bar (35 psi), the reaction mixture is filtered through Celite and washes the filter cake with water and ether. The aqueous phase is separated off, cooled to 0 ° C. and the pH is adjusted to 7.0 with saturated, aqueous NaHC03. After residual organic solvents have been removed in vacuo, the aqueous solution is added to a Cj8 reverse phase column. It is eluted with water and then with 8% acetonitrile-water and then the appropriate fractions are lyophilized, giving 0.25 g of a solid. This material is again purified by reverse phase HPLC, giving the product (0.114 g, 19%) as an off-white solid.

IR (KBr) v max: 3420 (OH), 1750 (β-lactam CO), 1590 (-COf) cm "1

tì-NMR (DiO) ô: 7.58 (s, 2H), 4.52 (t, J = 6 Hz, 2H), 4.28-3.82 (m, 2H), 3.90 (s, 3H, N-Me), 3.40-2.87 (m, 5H), 2.40 (s, 3H, S-Me), 1.20 (d, J = 6.4 Hz, 3H, -CHMe )

UV (H2Q) kmax: 297 (e 7572), 262 (e 6259), 222 (e 7955) nm

Example 12

Preparation of (5R, 6S) -3- [2- (3-aminopyridinio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2- en-2-car boxylate

SH.

30th

35

40

45

To a solution of p-nitrobenzyl- (5R, 6S) -6- [l- (R) -hydro-xyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2-carboxylate (0.696 g , 2.0 mmol) in 10 ml dry acetonitrile are added dropwise under N2 at 0 ° C diisopropylethylamine (0.382 ml, 2.2 mmol) and then diphenyl chlorophosphate (0.457 ml, 2.2 mmol), stirred for 30 min at 0 ° C , adds a solution of 3-amino-l- (2-mercaptoethyl) pyridinium chloride (0.475 g, 2.5 mmol) in 1 ml dry DMF and then further diisopropylethylamine (0.435 ml, 2.5 mmol), the reaction mixture is held 1.5 h at 0 ° C and then concentrated in vacuo. The rubbery product obtained is taken up in acetonitrile water (1: 1) and placed on a Ci8 reverse phase column. It is eluted with water, then with 20% acetonitrile-water and then the relevant fractions are lyophilized, the product (0.730 g, 50%) being obtained as a beige solid.

IR (KBr) v max: 3300 (br, OH), 3180 (br, NH2), 1770β-lactam CO), 1690 (-C02PNB) cm "1

! H NMR (d6DMSO) Ô: 8.29-7.63 (m, 8H, aromat.), 7.2-6.7 (m, 10H, diphenylphosphate), 5.47.5.18 (ABq, J = 14Hz, 2H, benzyl.), 4.73-4.45 (m, 3H), 4.2-3.8 (m, 1H), 3.6-2.6 (m, 8H), 1 , 15 (d, J = 6.2 Hz, 3H), CHMe).

C. (5R, 6S) -3- [2- (3-aminopyridinio) ethylIthio] -6- [l- (R) -hydro-xyethyl] -7-oxo-l-azabicyclo [3.2.0] hept- 2-en-2-carboxylate

50

55

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

O ^ "T> A

K. EC1

0

Ö

-i ©

3-aminopyridine (1.50 g, 0.016 mol) is taken up in 15 ml of 1N methanolic HCl, the solution obtained is evaporated off, with

60 To a mixture of p-nitrobenzyl- (5R, 6S) -3- [2- (3-aminopyridinio) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicy -clo [3.2.0] hept-2-en-2-carboxylate-diphenylphosphate (0.730 g, hh2 1.0 mmol) and 10% palladium-on-carbon (0.7 g) in 25 ml of a phosphate buffer (0 , 05 M, pH 7.4) 8 ml THF and 65 20 ml ether are added, the mixture is then hydrogenated for 1 h at a pressure of 2.76 bar (40 psi) (Parr), the mixture obtained is filtered through a pad of Celite and washes the filter cake with water and ether. The aqueous phase is separated off, washed twice with

29

658 248

Ether and then removes residual volatiles in vacuo. Immediately afterwards, the aqueous solution is placed on a C18 reverse phase column, which is eluted with water. Lyophilization of the elevation fractions gives 0.45 g of a dirty-white solid. This material is again purified by reverse phase HPCL, which gives the desired product (0.123 g, 35%) as an ivory colored solid.

IR (KBr) vmax: 3340 (br), 1750 (br, β-lactam CO), 1580 (br, -C02 ~) cm "" 1

iH-NMR (D20) ô: 8.07-7.59 (m, 4H, aromat.), 4.61 (t, J = 5.8 Hz, 2H, N-CH2), 4.14 (d of q, J = J '= 6.3 Hz, 1H, H-1'), 3.97 (d vont, J = 9.2 Hz, J '= 2.6 Hz, 1H, H-5), 3, 38 (t, J = 5.8Hz, 2H, S-CH2), 3.24 (d of d, J = 6.0Hz, J '= 2.6 Hz, 1H, H-6), 3.17- 2.57 (m, 2H, H-4), 1.21 (d, J = 6.3 Hz, 3H, CHMe)

UV (H20) Xmax: 299 (e 7949), 256 (s 8822) nm vA: (pH 7.4, 36.8 ° C) = 18.5 h.

Example 13

Preparation of (5R, 6S) -3- [l - ((S) -methyl-2- (l-pyridinium) -ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo - [3.2.0] hept-2-en-2-carboxylate

IR (film) vmax: 2500 (SH), 1628 (pyridinium), 1180 (sulfonate), 1035 (sulfonate) cm "1

H-NMR (DMSO-d6) ô: 1.69 (d, J = 6.8 Hz, 3H, CH3CHN +), 2.31 (s, 3H, CH3SO3-), 3.0-3.3 (m , 2H, CH2S), 4.2-5.2 (m, 1H, s CHN +), 8.0-8.4 (m, 2H, Hm of pyridinium), 8.5-8.8 (m, 1H , Hp of pyridinium), 9.0-9.2 (m, 2H, Ho of pyridinium) UV (H20) Xmax: 209 (e 4987), 258 (e 3838)

Analysis: calculated for C9H15N03S2-1.5H20: C 39.11%, H 6.56%, N 5.07%

10 found: C 39.13%, H 5.92%, N 5.20%

B. (5R, 6S) -p-nitrobenzyl-3- [1- (R, S) -methyl-2- (l-pyridinium) -

ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo-

[3.2.0] hept-2-en-2-carboxylate diphenyl phosphate

15 1) HEt (iPr),

Oh

A, _

"" JCM,,

Oh

A

20th

25th

COOPNB

3) rs

<> HEt (iPr) j ÖH ~

A ^ nv2 / K, ° "

A

eoo ©

COOPNB

(Pho) po "

and

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

OH

A

: of>

A. dl-l- (2-mercapto-2-methylethyl) pyridinium methanesulfonate dl-l- (2-mercapto-l-methylethyl) pyridinium methanesulfonate

A- "s

^ ± 1 + N

H MsOH

-Ol _

Methanesulfonic acid (1.95 ml, 0.030 mol) is slowly added to cold pyridine (7.83 ml, 0.097 mol), the mixture obtained is stirred for 15 min at 40 ° C., treated with dl-propylene sulfide (2.59 ml, 0.033 mol ) and stirred for 90 h under a nitrogen atmosphere at 60 ° C. The pyridine is removed in vacuo, the residue is mixed with water and purified by chromatography (HPLC, preparative Bondapak C-18). The appropriate fractions are combined and lyophilized to give dl-1- (2-mer-capto-2-methylethyl) pyridinium methanesulfonate (1.14 g, 15%) as a colorless syrup.

IR (film) vmax: 2520 (SH), 1640 (pyridinium), 1180 (st, CH3SO3-), 1040 (CH3SO3-) cm "1

XH-NMR (DMSO-d6) Ô: 1.35 (d, J = 6.8 Hz, 3H, CH3CHS), 2.30 (s, 3H, CH3SO3-), 2.90 (d, J = 8, 5 Hz, 1H, SH), 3.2-3.7 (m, CHSH), 4.52 (dd, Jgem = 12.9 Hz, J = 8.4Hz, CHCH2N +), 4.87 (dd, Jgem = 12.9 Hz, J = 6.0 Hz, CHCH2N +), 8.0-8.4 (m, 2H, Hm of pyridinium), 8.5-8.8 (m, 1H, Hp of pyridinium), 9.04 (dd, J = 1.4 Hz, J = 6.7 Hz, 2H, Ho of pyridinium)

UV (H20) Xmax: 208 (s 5267), 259 (s 3338)

Analysis: for C9Hi5N03S2-2H20 calculated: C 37.88%, H 6.71%, N 4.91%, S 22.47% found: C 37.49%, H 6.85%, N 4.86% , S 22.09%

Furthermore, dl-l- (2-mercapto-l-methylethyl) -pyridium-methanesulfonate (0.82 g, 11%) is obtained as a colorless syrup.

To a cold (0 ° C) solution of (5R, 6S) -p-nitrobenzyl-6- [l- (R) -hydroxyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2- Carboxylate (0.523 g, 1.5 mmol) in 6 ml acetonitrile (stored under a nitrogen atmosphere), diisopropylethylamine (0.314 ml, 1.8 mmol) and then diphenyl chlorophosphate (0.773 ml, 1.8 mmol) are added, the reaction mixture is stirred 30 min and treated with a solution of dl-1- (2- (mercapto-2-methyl-ethyl) -pyridinium methanesulfonate (0.539 g, 2.16 mmol) in 2 ml of 35 acetonitrile and diisopropylethylamine (0.314 ml, 1.8 mmol The reaction mixture is stirred for 1 h at 0 ° C., diluted with cold (0 ° C.) water (24 ml) and chromatographed on a prep. Bondapak C-18 column (2.5 × 8.5 cm), with 25 % to 50% acetonitrile in water was used as eluent After 40 lyophilization, 1.07 g (97%) of the title compound was obtained as a yellowish powder.

IR (KBr) vmax: 3700-3100 (OH), 1770 (C = 0 of ß-lactam), 1695 (C = 0 of the PNB ester), 1630 (pyridinium), 1590 (phenyl), 1518 (NO,) , 1348 (N02), 885 (N02) cm "" 1 45 1H-NMR (DMSO-d6) ô: 1.14 (d, J = 6.1Hz, 3H, CH3CHO), 1.33 (d, J = 6.3 Hz, 3H, CH3CHS), 4.6-5.0 (m, CH2N +), 5.14 (d, J = 5.2Hz, IH, OH), 5.37 (center of ABq, J = 12.4Hz, 2H, CH2 from PNB), 6.6-7.5 (m, lOH, phenyl of phosphate), 7.69 (d, J = 8.7 Hz, 2H, Ho from PNB), 8, 0-8.4 (m, 4H, Hm from PNB, Hm 50 from pyridinium), 8.4-8.8 (m, 1H, Hp from pyridinium), 9.08 (d, J = 5.6 H7, 2H, Ho of pyridinium)

UV (H20)> .max: 263 (e 13325), 308 (e 8915)

Analysis: calculated for C36H36N3Oi0SP-H2O: C 57.52%, H 5.10%, N 5.59%, S 4.27% found: C 57.76%, H 4.96%, N 5.36% , S 4.35%

55

60

65

C. (5R, 6S) -3- [l- (R and S) -methyl-2- (l-pyridinium) -ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l- azabicyclo [3.2.0] hept-2-en-2-carboxylate oh

"Vi

° COOPNB (PhO) 2PO

THF, ether, O buffer

0h

A

coo

658 248

30th

To a solution of (5R, 6S) -p-nitrobenzyl-3- [l- (R, S) -methyl-2- (l-pyridinium) -ethylthio] -6- [l- (R) -hydroxyethyl] - 7-oxo-l-azabicyclo [3.2.0] hept-2-en-2-carboxylate-diphenylphosphate (0.60 g, 0.82 mmol) in moist tetrahydrofuran (33 ml)

are added ether (33 ml), potassium phosphate monobasic 5

Sodium hydroxide buffer (17 ml, 0.15 M, pH 7.22) and 10% palladium-on-carbon (0.60 g) and hydrogenated the resulting mixture for 1 h at 23 ° C. and a pressure of 2.76 bar (40 psi). The two layers are separated and the organic layer extracted with water (3 x 7 ml). The aqueous layers are combined, filtered through a pad of Celite, washed with ether (3 X20 ml) and chromatographed on a prep. Bondapak C-18 column (2.5x9.5 cm), using water as the elution solvent. 0.18 g (63%) of a mixture of diastereoisomers is obtained. The two diastereoisomers are separated by HPLC (prep. Bondapak C-18), water being used as the eluent. 0.068 g (23%) of the isomer with the shorter retention time, referred to as compound «B», is obtained.

IR (KBr) v max: 1770 (C = 0 from β-lactam), 1633 (pyridinium), 20 1593 (carboxylate) cm "1

^ -NMR (D20) ô: 1.20 (d, J = 6.3 Hz, 3H, CH3CHO), 1.42 (d, J = 6.9 Hz, 3H, CH3CHS), 2.3-3, 2 (m, 3H, H-4, H-6), 3.5-3.9 (m, IH, SCH), 3.9-4.2 (m, 2H, H-5, CH3CHO), 4 , 3-5.1 (m, CH2N ") 7.8-8.2 (m, 2H, Hm of pyridinium), 8.4-8.7 (m, lH, Hp of pyridinium), 8.7-9 , 0 (m, 2H, Ho of pyridinium)

UV (H20) Xmax: 260 (s 6727), 300 (e 8245)

[ag = -39.3 ° (c, H20)

T! / ,: 12.6 h (measured at a concentration of 10 "" 4 M in phosphate buffer pH 7.4 at 36.8 ° C).

0.081 g (28%), designated compound “A”, is also obtained as the isomer with a longer retention time.

IR (KBr) v max: 1755 (C = O of β-lactam), 1630 (pyridinium), 1590 (carboxylate) cm-1

XH-NMR (D20) ô: 1.18 (d, J = 6.3Hz, 3H, CH3CHO), 1.40 35 (d, J = 7.0 Hz, 3H, CH3CHS), 2.84 (d, J = 9.3 Hz, 2H, H-4), 3.26 (dd, J = 2.7 Hz, J = 5.9 Hz, 1H, H-6), 3.4-4.2 (m , 3H, SCH, CH3CHO, H-5), 4.2-5.1 (m, CH2N +), 7.7-8.1 (m, 2H, Hm of pyridinium), 8.3-8.65 ( m, 1H, Hp of pyridinium), 8.65-8.9 (m 2H, Ho of pyridinium)

UV (H20) Xmax: 259 (e 5694), 296 (e 6936)

[a] n = + 96.9 ° (£ 0.56, H20)

15.6 h (measured at a concentration of IO-4 M in phosphate buffer pH 7.4 at 36.8 ° C)

A. dl-l- (2-mercapto-l-cyclohexyl) pyridinium methanesulfonate see

Q> + «o) + msoh - * OC N-

MsO

25th

30th

Methanesulfonic acid (0.65 ml, 0.01 mol) is added dropwise with cooling to pyridine (2.42 ml, 0.03 mol), the mixture is stirred for 10 min under a nitrogen atmosphere, treated with dl-cyclohexene sulfide [1.377 g (85 % purity), 0.0102 mol) and stirred for 25 h at 72 ° C. Excess pyridine is removed in vacuo. Traces are distilled together with water. The residue is mixed with water and chromatographed on a prep. Bondapak C-18 column (5 X13 cm) with 0-2% acetonitrile in water as eluent. After lyophilization, 1.57 g (53%) of a colorless syrup is obtained.

IR (film) vmax: 2500 (SH), 1625 (pyridinium), 1190 (S03 ") 1H-NMR (DMSO-d6) Ô: 1.2-2.5 (m, 8H, cyclohexylH), 2.32 ( s, 3H, CH3SO3-), 2.82 (d, J = 9.8 Hz, SH), 3.0-3.5 (m, 1H, CHSH), 4.2-4.9 (m, 1H , CHN +), 8.0-8.3 (m, 2H, Hm of pyridinium), 8.4-8.8 (m, 1H, Hp of pyridinium), 8.9-9.3 (m, 2H, Ho of pyridinium)

UV (H20) 214 (e 5365), 258 (e 3500)

Analysis: calculated for Ci2HI9N03S2-H20: C 46.88%, H 6.88%, N 4.56%

found: C 46.61%, H 6.46%, N 4.65%

B. (5R, 6S) -p-nitrobenzyl-3- [2 - [(R or S) - (l-pyridinium)] - l- (R orS) -cyclohexylthio] -6- [l- (R) - hydroxyethyl] -7-oxo-l-azabi-cyclo [3.2.0] hept-2-en-2-carboxylate diphenyl phosphate

OH

A

ro-

1) NEt (iPr) 2

P

2) ClP (OPh)

3)

"TD

«> NEt (iPr),

40

A

45

COOPNB (PhO) -PO '

[gj

Example 14

Preparation of (5R, 6S) -3- [2 - [(S) - (l-pyridinium)] - l- (S) -cyclo-hexylthio] -6- [l- (R) -hydroxyethyl] -7- oxo-l-azabicyclo [3.2.0] hept-2-en-2-carboxylate

OH

A

.-O

and

(5R, 6S) -3- [2- (R) - (l-pyridinium-l- (R) -cyclohexylthio] -6- [l- (R) -hydroxyethylj-7-oxo-l-azabicyclo [3.2. 0] hept-2-en-2-carboxylate

OH

k

To a cold (0 ° C) solution of (5R, 6S) -p-nitrobenzyl-6- [l- (R) -hydroxyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2- Carboxylate (1.37 g, 3.93 mmol) in 15 ml acetonitrile (stored under a nitrogen atmosphere) are added diisopropylethyl-50 amine (0.822 ml, 4.7 mmol) to diphenyl chlorophosphate (0.979 ml, 4.7 mmol). The solution obtained is stirred for 30 min, treated with a solution of dl-l- (2-mercapto-l-cyclohexyl) pyridinium methanesulfonate (1.64 g, 5.66 mmol) in 4.7 ml of acetonitrile and then with Diisopropylethylamine (0.822 ml, 55 4.7 mmol). The reaction mixture is stirred for 1 h at 0 ° C., diluted with cold (0 ° C.) water (75 ml) and chromatographed using a prep. Bondapak C-18 column using 25-50% acetonitrile in water as the eluent. After lyophilization of the appropriate fractions, 1.9 g of 60 (53%) of the title compound are obtained.

IR (KBr) vmax: 3700-3000 (OH), 1770 (C = 0 of ß-lactam), 1700 (C = 0 of the PNB ester), 1628 (pyridinium), 1590 (phenyl), 1515 (NO,) , 1345 (NO,), 880 (NO,) cm "1

'H NMR (D, 0) Ô: 1.13 (d, J = 6, l, H7.3H, CH3CHO), 651.2-2.5 (m, 3H, cyclohexyl H), 2.7- 3.5 (m, 4H, H-4, CHS), 3.5-4.4 (m, 2H, CH3CHO, H-5), 4.4-5.0 (m, 1H, CHN +), 5 , 30 (center of ABq, J = 12.8 Hz, CH, of PNB), 6.7-7.4 (m, lOH, phenyl), 7.65 (d, J = 8.6 Hz, 2H, Ho from PNB), 7.9-8.4 (m,

31

658 248

4H, Hm from PNB, Hm from pyridinium), 8.4-8.8 (m, 1H, Hp from pyridinium), 9.0-9.4 (m, 2H, Ho from pyridinium) UV (H20) lmax: 263 (e 9038), 309 (e 6394)

Analysis: for C39H4oN3010SP • H20 calculated: C 59.16%, H 5.35%, N 5.31%

found: C 58.95%, H 5.15%, N 5.57%

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

10% wc, h,

10th

UV (H20)> .max: 259 (e 5992), 296 (e 7646)

[cx] d = + 65.30 (_c 0.43, H20)

ziA: 20.2 h (measured at a concentration of 10 "4 M in phosphate buffer pH 7.4 at 36.8 ° C)

Example 15

A. (5R) -Allyl-3 - [(2-pyridinioethyl) thio] - (6S) - [(IR) -hydroxy-ethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2- en-2-carboxylate diphe nyl phosphate

OH

A

THF, ether, buffer

1) «Poloni

15

20th

Oh

C02 ^ -bPOfOM

To a solution of (5R, 6S) -p-nitrobenzyl-3- [2 - [(R or S) - (l-pyridinium)] - l- (R or S) -cyclohexylthio] -6- [l- ( R) -hydro-xyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2-carboxylate-diphenylphosphate (1.85 g, 2.34 mmol) in moist tetrahydrofuran (96 ml ) ether (96 ml), potassium phosphate-monobasic sodium hydroxide buffer (0.15 M, pH 7.22.50 ml) and 10% palladium-on-carbon (1.9 g) are added and the mixture obtained is hydrogenated 1, 25h at 23 ° C under a pressure of 2.76bar (40 psi). The organic layer is separated off, washed with water (3 × 20 ml), the aqueous solutions are filtered through a pad of Celite, washed with ether (2 × 60 ml) and traces of organic solvents are removed by pumping and chromatographed on a prep. Bondapak C-18 column (4.5x9 cm) with 0-5% acetonitrile in water as eluent. After lyophilization, 0.705 g (76%) of a mixture of diastereoisomers is obtained. These diastereoisomers are separated by means of HPLC (prep. Bondapak C-18) with 4% acetonitrile in water as the eluent. The diastereoisomer with the lower retention time is designated compound “A” (0.29 g, 31%).

IR (KBr) vmax: 1750 (C = 0 of β-lactam), 1620, (Sch, pyridinium), 1685 (carboxylate) cm "1

1H NMR (D20) ô: 1.21 (d, J = 6.3 Hz, 3H, CH3CHO), 1.4-2.5 (m, 3H, cyclohexylH), 2.5-3.05 (m , 2H, H-4), 3.05-3.25 (m, 1H, H-6), 3.3-3.7 (m, 1H, CHS), 3.9-4.3 (m, 2H, H-5, CHjCHO), 4.3-4.8 (m, CHN +), 7.8-8.2 (m, 2H, Hm of pyridinium), 8.3-8.7 (m, 1H , Hp of pyridinium), 8.8-9.1 (m, 2H, Ho of pyridinium)

UV (H20)> "max: 260 (e 7123), 300 (e 8685)

[a] n = + 6.2 ° (_c 0.63, H20)

ziA \ 16.6 h (measured at a concentration of 10 "4 M in phosphate buffer pH 7.4 at 36.8 ° C)

Analysis: calculated for C20H24N2O4S-2H2Ó: C 56.59%, H 6.65%, N 6.60%, S 7.55% found: C 56.83%, H 6.47%, N 6.59% , S 7.43% The diastereoisomer with the higher retention time is referred to as compound «B» (0.35 g, 38%).

IR (KBr) vmax: 1750 (C = Ovon β-lactam), 1622 (Sch, pyridinium), 1588 (carboxylate) cm "1

1H-NMR (D20) Ô: 1.19 (d, J = 6.4Hz, 3H, CH3CHO), 1.3-2.5 (m, 8H, cyclohexylH), 2.5-3.1 (m, 2H, H-4), 3.1-3.3 (m, 1H, H-6), 3.3-3.8 (m, 2H, H-5, CHS), 4.1 (center of m, IH, CH3CHO), 4.25-4.7 (m, 1H, CHN +), 7.8-8.1 (m, 2H, Hm of pyridinium), 8.3-8.7 (m, 1H, Hp of pyridinium), 8.75-9.0 (m, 2H, Ho of pyridinium)

To a solution of (5R) -Allyl-3,7-dioxo- (6S) - [(IR) -hydro-xyethyl] -l-azabicyclo [3.2.0] heptane (2R) carboxylate (473 mg, 1 , 87 mmol) in 6 ml of CH3CN, diisopropylethylamine (0.42 ml, 2.4 mmol) is added at about -10 ° C. under a nitrogen atmosphere and then diphenyl chlorophosphate (0.50 ml, 2.4 mmol). The mixture is stirred for 30 min at 0 ° C, cooled to -15 ° C, gives an oily suspension of N- (2-mercaptoethyl) pyridinium chloride (527 mg, 3.00 mmol) in 1 ml of CH3CN containing of 5 30 drops of DMF and then diisopropylethylamine (0.42 ml, 2.4 mmol). The mixture is stirred for 30 min at -15 ° C, diluted with 20 ml of water, this mixture is purified directly on a reversed-phase silica gel column (C18PrepPAK, 12 g, Waters Associates) while eluting with water (200 ml) 10% CH3CN / H20 35 (100ml), 20% CH3CN ° CH20 (100ml), 30% CH3CN / H20 (100 ml) and then with 40% CH3CN / H20 (100 ml). The appropriate fractions are collected, the organic solvent is removed with the aid of a vacuum pump and lyophilized, giving 786 mg (1.26 mmol, yield 67.3%) of the title compound as a brownish powder.

• H NMR (DMSO-d6, CFT-20) Ô: 1.16 (3H, d, J = 6 Hz, 1'-CH3), 2.6-3.7 (m), 3.75-4 , 3 (2H, m, 5-H, and l'-H), 4.65 (2H, m, -C02CH2-), 4.87 (2H, t, J = 6 Hz, -CHr-N +), 5-6.2 (3H, m, olefinic protons), 6.6-7.4 (m, aromatic protons), 8.15 (2H, 451, J = 7Hz, aromatic protons meta to nitrogen), 8 , 63 (1H, t, J = 7 Hz, aromatic proton para to nitrogen), 9.07 ppm (2H, d, J = 7 Hz, aromatic proton ortho to nitrogen)

IR (film) vmax: 3400 (OH), 1770 (β-lactam), 1690 (ester), 1625 (pyridinio).

50

B. (5R) -3 - [(2-pyridinioethyl) thio] - (6S) - [(IR) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2- carboxylate

OH

A,

55

60

0J2>

©,

Pd (P $ 3) 4

p $,

CO ®opo (o $)

OH

A,

To a solution of (5R) -Allyl-3 - [(2-pyridinioethyl) thio] -65 (6S) - [(IR) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2 -en-2-carboxylate-diphenylphosphate (156 mg, 0.25 mmol) in 2 ml of CH3CN, a solution of potassium 2-ethylhexanoate in EtOAc (0.5 M, 0.6 ml ; 0.3 mmol),

658 248

32

Triphenylphosphine (15 mg, 0.057 mmol) and tetrakistriphenyl-phosphine-palladium (15 mg, 0.013 mmol) and the mixture is stirred for 2 hours at approx. 22 ° C. under a nitrogen atmosphere. After adding 7 ml of anhydrous Et20, the precipitate is filtered, washed with 7 ml of anhydrous Et20 and dried in vacuo to give 101 mg of a brownish solid. This is purified by reverse phase column chromatography (Cjg PrepPAK, 12 g, Waters Associates) while eluting with water. Suitable fractions (Fr.7-12, 20 ml each) are collected and lyophilized to give 53 mg (0.16 mmol, yield 64%) of the title compound as a yellowish powder. This material is contaminated with potassium diphenyl phosphate and potassium 2-ethylhexanoate.

'H NMR (D20, CFT-20) Ô: 0.80 (t, J = 6.4Hz, Me from ethylhexanoate), 1.21 (3H, d, J = 6.3 Hz, l'-Me) , 2.93 (2H, dd, J1_5 = 9 Hz, Jgem = 4 Hz, 1-s), 3.28 (1H, dd, JM = 6.2Hz, JM = 2.5 Hz, 6-H), 3.42 (2H, t, J = 6 Hz, -CH2S), 3.98 (1H, td, 1 ^ = 9 Hz, 15.6 = 2.5 Hz, 5-H), 4.15 (1H, q , J = 6.2Hz, l'-H), 4.80 (2H, t, J = 6.0 Hz, -CH2N +), 7-7.5 (m, phenyl protons from diphenyl phosphate), 8.03 (2H , m, Hm of pyridinium), 8.56 (1H, m, Hp of pyridinium) and 8.81 ppm (2H, d, J = 6.5 Hz, Ho of pyridinium).

A cold (ice bath) solution of p-nitrobenzyl-6a- [l '(R) -hydroxyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2-carboxylate (557 mg, 1.60 mmol ) in 8 ml of CH3CN, dropwise treated with diisopropylethylamine (0.336 ml, 1.92 mmol) and diphenyl chlorophosphate (0.400 ml, 1.92 mmol), stirred for 30 min, the reaction mixture was treated with N-methyl-N- (2-mercaptoethyl) -thio-morpholinium methanesulfonate (893 mg, 2.29 mmol) in 4 ml CH3CN and with diisopropylethylamine (0.336 ml, 1.92 mmol) and stirred for 30 min. The solution is diluted with 20 ml of water and poured onto a silica gel reverse phase column. The desired compound is eluted with a 50% acetonitrile-water mixture, the appropriate fractions are combined, pumped under vacuum for 2 h and lyophilized to give the title compound (1.01 g, 85%).

IR (Nujol) vmax: 1760 (s, β-lactam C = 0), 1510 cm "1 (st, N02) cm" 1

! H NMR (DMSO-d6) Ô: 8.25 (2H, d, J = 8.8 Hz, H-aromat.), 7.70 (2H, d, J = 8.8 Hz, H-aromat .), 7.33-6.84 (10H, m, H-aroma), 5.37 (2H, center of ABq, J = 14.2Hz, CH2), 5.14 20 (1H, d, J = 4.5 Hz, OH), 4.35-3.80 (2H, m, Hl 'and H-5), 3.75-3.45 (6H, m, CH, N +), 3.31 ( 3H, s, CH3N +), 3.45-2.75 (9H, m, CH-.S, H-6 and H-4) and 1.15 ppm (3H, d, J = 6.2 Hz, CH3 ).

10th

15

Example 16

Preparation of 3- [2- (N-methylthiomorpholinium) ethylthio] -6a- [l '- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2 carboxylate

25 C. 3- [2- (N-methylthiomorpholinium) ethylthio] -6a- [l '- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en- 2-carboxylate

30th

X

ri

® i

CO.ÏNB

^ 0 = fOÇ) 2

A.N-methyl-N- (2-mercaptoethyl) thiomorpholinium methane sulfonate

-O

1) MSOH

Hey / \

2) Â

To pre-cooled (ice bath) N-methylthiomorpholine (5.00 g, 42.7 mmol) [s. J.M. Lehn and J. Wagner, Tetrahedron, 26.4227 (1970)] are added methanesulfonic acid (1.47 ml, 20.5 mmol) and ethylene sulfide (1.30 ml, 21.4 mmol), the mixture is heated to 65 for 24 h ° C and diluted with 25 ml of water. The aqueous solution is washed with diethyl ether (3 x 25 ml), pumped under vacuum and poured onto a silica gel reverse phase column. The title compound is eluted with water. The appropriate fractions are combined and concentrated to give the thiol as an oil (4.80 g, yield 86%).

IR (film) vmax: 2550 cm "1 (black SH),

! H NMR (DMSO-de) Ô: 3.25-2.95 (6H, m, CH2N +), 3.32 (3H, s, CH3N +), 3.20-2.65 (7H, m, CH2S , SH) and 2.32 ppm (3H, s, CH3S03).

B. p-Nitrobenzyl-3- [2- (N-methylthiomorpholinium diphenylphosphate) ethylthio] -6a- [l '- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2 .0] hept-2-en-2-carboxylate

OH D EtN «- <> 2

2). C1? 0 ($ 0) 2

35

40

OH

X,

G

© o

X

3)

3) HS-v ^, n

Hey - EtN <- <),

** '- 5 He / V

s,

O-- - © \ /

1 <3

CO-PNB OP (O0) _

A solution of p-nitrobenzyl-3- [2- (N-methylthiomorpho-linium diphenylphosphate) ethylthio] -6a- [l '- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2. 0] Hept-2-en-2-carboxylate (1.31 g, 1.76 mmol) in 0.1M pH 7.4 phosphate buffer (48.8 ml), tetrahydrofuran (20 ml) and diethyl ether (20 ml) hydrogenated one hour at 45 2.76 bar (40 psi) over 10% Pd / C (1.5 g) in a Parr shaker. The reaction mixture is diluted with 40 ml of diethyl ether and the phases are separated. The organic phase is washed with water (2x5 ml), the aqueous phases are combined, filtered through a No. 52 hardened filter paper, washed with diethyl ether (2x20 ml) and a vacuum is applied. The aqueous solution is poured onto a silica gel reverse phase column and the desired carbapenem is eluted with 5% acetonitrile water. The appropriate fractions are combined and lyophilized to give the title compound as an amorphous solid (205 mg, 31%).

IR (Nujol) vmax: 1750 (st, ß-lactam C = 0), 1590 cm "1 (st, C = 0)

H-NMR (D, 0) ò: 4.25-3.95 (2H, m, H-l ', H-5), 3.70-3.40 (6H, m, CHiN +), 3, 55 (1H, dd, J = 6, 1 Hz, J = 2.6 Hz, H-6), 3.08 (3H, s, CH3N +), 3.25-2.75 (8H, CH, S, H-4) and 1.24 ppm (3H, 60 d, J = 6.4 Hz, CH3)

UV (HoO; 0.062) Xmax: 299 (e 10962)

xi,: 17.7 h (0.1 M pH 7 phosphate buffer, 37 ° C).

50

55

65 Example 17

Preparation of (5R, 6S) -3- [2- (l-methylmorpholino) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2- en-2-carboxylate

33

658 248

OH

A,

A. l-Methyl-l- (2-mercaptoethyl) morpholinium trifluoromethanesulfonate

To a solution of p-nitrobenzyl- (5R, 6S) -3- [2- (l-methylmorpholino) ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabi- Cyclo [3.2.0] hept-2-en-2-carboxylate-diphenylphosphate (0.360 g, 0.49 mmol) in 13 ml of phosphate buffer (0.05 M pH 7.4) are added

0 + cf3so3h + / v ^ hschjchj / \ io 0.36 g 10% palladium-on-carbon, 20 ml tetrahydrofuran and c 20 ml diethyl ether and hydrogenated this mixture (Parr) for 1 h under me cf3so3 at a pressure of 2.2 bar (32 psi). The mixture is filtered

To N-methylmorpholine (3.29 ml, 0.030 mol) is added through Celite, the filter pad is washed with water and diethyl-10 ° C. dropwise trifluoromethanesulfonic acid (1.327 ml, ether), the aqueous phase is separated off and the pH is adjusted

0.015 mol) and then ethylene sulfide (0.89 ml, 0.015 mol), 15 additional pH 7.4 phosphate buffer to 7.0. After removal, the resulting yellow-brown solution is heated for 18 hours under N2 on the residual organic solvent in vacuo, an oil bath is added at 50 to 60 ° C., then volatile material in the aqueous solution is removed on a C18 reverse phase column. Elution

Vacuum and take up the remaining oil in 10 ml of water. The aqueous fractions obtained with water and lyophilization of the relevant fractions are washed with diethyl ether (3x5 ml), 0.130 g of an amorphous solid is removed. This material is then cleaned of residual organic solvents in vacuo and the 20 are added again by reverse phase HPLC, the pure resulting aqueous solution on a C18 reverse phase column, product (0.058 g, 34%) being obtained as an amorphous solid.

which one with water, then 5% acetonitrile-H20 and finally IR (KBr) vmax: 3420 (br, OH), 1750 (ß-lactam CO), 1590

10% Acetonitrile-H20 eluted. Evaporation of the relevant Frak- (-C02 ~), cm-1

ions gives a white solid which is dried in vacuo 'H-NMR (D20) ô: 4.35-2.77 (m, 17H), 3.18 (s, 3H, N-Me) (P205), whereby the product is obtained (1.92 g, 41%). 25 1.23 (d, J = 6.3 Hz, 3H, CHMe)

UV (H20) Xm, x: 300 (e 6344) nm (pH 7.4, 36.8 ° C) = 18.5 h.

IR (KBr) vmax: 2560 (-SH) cm "1 MMR (d6-acetone) Ô: 4.25-3.6 (m, 8H), 3.49 (s, 3H, N-Me), 3, 35-2.7 (m, 5H)

Bp-nitrobenzyl- (5R, 6S) -3- [2- (l-methylmorpholino) ethylthio] -6 - [(R) -l-hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept-2-en-2-carboxylate diphenyl phosphate

OH

0 coopnb

OH

A,

//

Me .

\ / 35

O

II

Example 18

30 Preparation of (5R, 6S) -3- [2- (1,4-dimethyl-l-piperazinium) -ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo- [ 3.2.0] hept-2-en-2-carboxylate

CH3 / v

'(00) 2po °

To a solution of p-nitrobenzyl- (5R, 6S) -6 - [(R) -l-hydro-xyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2-carboxylate (0.348 g , 1.0 mmol) in 25 ml of dry acetonitrile are added dropwise diisopropylethylamine (0.191 ml, 1.1 mmol) and then diphenylchlorophosphate (0.228 ml, 1.1 mmol) at 0 ° C. under N 2, after stirring for 1 hour at 0 ° C diisopropylethylamine (0.226 ml, 1.3 mmol) to the resulting enolphosphate and then l-methyl-l- (2-mercaptoethyl) morpholinium trifhioromethanesulfonate (0.373 g, 1.2 mmol). The reaction mixture is stirred for 1.5 h at room temperature, concentrated in vacuo, the remaining material is taken up in water and eoo

40

45

A. 1- (2-Acetylthioethyl) -1, 4-dimethylpiperazinium bromide

N-, 3r

A solution of 2-bromoethyl thiol acetate (2.20 g, 0.012 mol) [s. B. Hansen, Acta Chem. Scand. 11.537-40 (1957)] and 1,4-dimethylpiperazine (1.95 ml, 0.014 mol) in 4 ml of acetone are stirred onto a C18 reverse phase column. Elution with water, then 50 65 h at 50 ° C. After cooling to 25 ° C., the 20% acetonitrile-H20 and finally 30% acetonitrile-H20 and liquid phase are decanted from the rubber-like material, which is obtained by twice subsequent lyophilization of the relevant fractions, and triturated with diethyl ether, giving a hygroscopic, yellow one Product (0.360 g, 40%) as an amorphous solid. Lich powder obtained (3.2 g, 90%).

IR (film) vmax: 3300 (-OH), 1770 (β-lactam CO), 1700 (IR (Nujol) vraax: 1685 (C = 0 of the thioester) cnT1

(-C02PNB) cm "1 55! H NMR (D20) Ô: 2.37, 2.39 (2s, 6H,

'H NMR (d6-acetone) Ô: 8.25.7.80 (ABq, J = 8.6 Hz, 4H,

arom.), 7.4-6.8 (m, 10H, diphenylphosphate), 5.56.5.27 (ABq, J = 14.2Hz, 2H, benzyl.), 4.42 (dvont, J = 9 , 2Hz, J '= 2.7Hz, 1H, H-5), 4.1-2.7 (m, 17H), 3.40 (s, 3H, N-Me), 1.22 (d, J = 6.2 Hz, 3H, -CHMe).

C. (5R, 6S) -3- [2- (l-methylmorpholino) ethylthio] -6 - [(R) -l-hydroxyethyl] -7-oxo-l-azabicyclo [3.2.0] hept- 2-en-2-carboxylate

X. ^ v- \

0

CHjCO,

/ -CHj), 3.18 (s, 3H

-cn3

).

60 B. 1,4-dimethyl-l- (2-mercaptoethyl) -piperaziniumbromid-hy-

hydrochloride

, N N—, Br w

? n3 8

65

HCl.

A

e

KCl

658 248

34

A solution of l- (2-acetylthioethyl) -1, 4-dimethylpiperazi-nium bromide (1.1 g, 3.7 mmol) in 6N hydrochloric acid (4 ml) is heated for 1 h at 80 ° C. under a nitrogen atmosphere and the mixture is concentrated Solution under reduced pressure to give a white powder (0.41 g, 38%).

H-NMR (DMSO-de) ô: 2.90 (s, "V *"), 3.26 (s,

v-%

• CHj

)

Analysis: for C8H20N2SBrCl-H2O

calculated: C 31.03%, H 7.16%, N 9.05%, S 10.35%

found: C 31.62%, H 7.46%, N 9.19%, S 10.19%

C. (5R, 6S) -p-nitrobenzyl-3- [2- (1,4-dimethyl-l-piperazinium) -ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l- azabicyclo- [3.2.0] hept-2-en-2-carboxylate-diphenylphosphate qjj 1J HEt (iPr) j

«

I \ 2) ClP (OPh) 2 ^

= o-

Br, HCl

4) NEtdPrïj

Jf-.

COOPBB (Ph0) 2P0-

To a cold (0 ° C) solution of (5R, 6S) -p-nitrobenzyl-6- [l- (R) -hydroxyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2- (R) -carboxylate (0.465 g, 1.33 mmol) in 2 ml of acetonitrile (stored under a nitrogen atmosphere) are added diisopropylethylamine (0.278 ml, 1.59 mmol) and diphenyl chlorophosphate (0.33 ml, 1.59 mmol), the reaction mixture is stirred for 30 min and treated with a suspension of 1,4-dimethyl-1- (2-mercap-toethyl) piperazinium bromide hydrochloride (0.40 g, 1.37 mmol) in 3 ml of acetonitrile-1 ml of water. Mixture and diisopropylethylamine (0.278 ml, 1.59 mmol). After stirring at 5 ° C. for 18 hours, 15 ml of cold water are added to the mixture and the resulting solution is chromatographed on a PrePak-500 C18 (Waters Associates) column (2.5 × 7.5 cm), with 25-35% acetonitrile in Water used as eluent. After lyophilization, a yellowish powder (0.50 g, 50%) is obtained.

IR (KBr) v max: 1765 (C = O of β-lactam), 1690 (c = 0 of PNB ester), 1585 (phenyl), 1512 (N02), 875 (N02) cm "1

! H NMR (DMSO-d6) ô: 1.16.1.18 (2d, J = 6, 1 Hz, 3H, CH3CHOH), 2.44,

(s, N-CH3), 3.14 (a.

• CH,

-3),

To a solution of (5R, 6S) -p-nitrobenzyl-3- [2- (1,4-dimethyl-l-piperazinium) -ethylthio] -6- [l- (R) -hydroxyethyl] -7- oxo-l-azabicyclo [3.2.0] hept-2-en-2-carboxylate-diphenylphosphate

(0.47 g, 0.623 mmol) in 25 ml of moist tetrahydrofuran are added diethyl ether (25 ml), potassium phosphate-monobasic sodium hydroxide buffer (13 ml, pH 7.22) and 10% palladium-on-carbon (0.47 g ), the mixture obtained is hydrogenated for 1 h at 23 ° C. 5 under a pressure of 2.76 bar (40 psi), separates the two layers and extracts the organic layer with water (2x7 ml). The aqueous layers are combined, filtered through a Celite cushion, washed with diethyl ether (2x15 ml) and chromatographed on a PrePak-500 / C18 (Waters Associates) -10 column (2.5x9.5 cm) with water as the eluent, where 0.097 g (43%) of product is obtained after lyophilization.

IR (KBr) vmax: 3000-3700 (OH), 1750 (C = 0 of β-lactam), 1585 (carboxylate) cm "1

'H NMR (D20) ô: 1.24 (d, J = 6.4 Hz, 3H, CH3CHOH), 2.33

15

(s, 3H, y -'- CHj), 3.15 (s,

-V '-—CHS

N + 3),

4.0-4.5 (m, H-5, CH3CHOH)

UV (H20) Àmax: 296 (e 9476)

[a] 2D3 = 61, r (C 0.26, H, 0)

20 ti ^: 12.4 h (measured at a concentration of 10 "4 M in pH 7.4 phosphate buffer at 36.8 ° C).

Example 19

25 Preparation of (5R, 6S) -3- [2- (N-methylthiomorpholini-umoxid] -ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo- [3.2. 0] hept-2-en-2-carboxylate

30th

35

40

PQr

C02e

OK

A,

ST \

«<\ _ V

45

50

5.31 (d, J = 6 Hz, OH), 5.39 (center of ABq, J = 13 Hz, CH, of PNB), 6.6-7.4 (m, lOH, phenyl of phosphate), 7.71 (d, J = 8.8 Hz, 2H, Ho from PNB), 8.26 (d, J = 8.8 Hz, Hm from PNB).

D. (5R, 6S) -3- [2- (1,4-dimethyl-l-piperazinium) -ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2. 0] hept-2-en-2-carb-oxylate

► v r ~ \ 0

, Ph0, J? 0 "

é? N ^ cooPNa

55

60

65

To a cold (-10 ° C) solution of (5R, 6S) -3- [2- (N-methylthiomorpholinium) ethylthio] -6- [l- (R) -hydroxyethylj-7-oxo-l- azabicyclo [3.2.0] hept-2-en-2-carboxylate (608 mg, 1.65 mmol) in a 1: 1 mixture of acetonitrile-water (9 ml) is added in small portions of m-chloroperbenzoic acid for 1 h (334.8 mg, 1.65 mmol), then dilute the mixture with 15 ml of water, wash with diethyl ether (3x15 ml), treat the aqueous phase in vacuo and pass over a reverse phase silica gel column (H20) to give a solid receives, which consists of a mixture of compounds. This mixture is separated by reverse phase HPLC, giving fraction A (52.4 mg; yield 12%) and fraction B (23.6 mg; yield 6%) as diastereoisomers of the title compound.

Fraction A

IR (Nujol) vmax: 1750 (st, ß-lactam C = 0), 1580 cm "1 (st, C = 0)

H-NMR (D-.O) Ô: 4.26-2.91 (20H, m, H-4, H-5, H-6, H-1 ', CH-.S, CHnS-O, CH3-N + and CH, M +) and 1.24 ppm 3H, d, J = 6.4Hz, CH,)

UV (H20) /.mM: 302 (e 10425)

Ti-w 12 h (0.065 M, pH 7.4 phosphate buffer, 37 ° C.

Fraction B

IR (Nujol) vmax: 1750 (st, ß-lactam C = 0) and 1585 cm "1 (st, C = 0)

35

658 248

^ -NMR (D20) ô: 3.86-2.90 (1H, m, H-4, H-5, H-6, H-l ', CH2S, CH2S-0, CH2N +), 3.25 ( 3H, s, CH3N +) and 1.24 ppm (3H, d, J = 6.4 Hz, CH3)

UV (H20, c 0.05) amax: 299 (s 6517)

Xi / 2-10.75 h (0.065 M, pH 7.4 buffer solution, 37 ° C).

Example 20

Preparation of (5R, 6S) -3- [2- (l, 4,4-trimethyl-l-piperazinium) -ethylthio] -6- [l- (R) -hydroxyethyl] -7-oxo-l-azabicyclo- [3.2.0] hept-2-en-2-carboxylate chloride cr

C. (5R, 6S) -p-nitrobenzyl-3- [2- (l, 4,4-trimethyl-l-piperazionium) ethylthio] -6- (lR) -hydroxyethyl] -7-oxo-l -azabicyclo- [3.2.0] hept-2-en-2-carboxylate-bis-chloride

OH

A

O

•• COOPKB

1) NEC (iPr) 2

2) Ci? (OPh) 2

Permutite S-1 Cl

3) NCtUPr) A) KS «^ S> s CH

2C1 ~

! \

10th

A

> / rv

'COOPNB

7 \ + 2c1 "

'coo

A. l- (2-acetylthioethyl) -l, 4,4-trimethylpiperaziniumbromid-

iodide s / \ _ - K «J

ch.

& '

'W

^ -OH, 55-

■ £.

\ f ~ \ /. .

X N + * v +. E =. J

V_ / \

A suspension of 1- (2-acetylthioethyl) -1, 4-dimethylpipe-razinkum bromide (1.48 g, 5.0 mmol) in 10 ml of isopropyl alcohol is treated with methyl iodide (0.373 ml, 6.0 mmol) and heated 30 h to 55 to 60 ° C. The solvents are evaporated off under reduced pressure, the residue is triturated in hexane and the solid is filtered (1.85 g). The solid is dissolved in 8 ml of hot water, the solution is diluted until cloudy with acetone (70-80 ml) and two successive crystallizations are carried out, 1.5 g (68%) of the title compound, mp. 220 to 225 ° C (dec.).

IR (KBr) vmax: 1692 cm "1 (C = 0)

'H NMR (D20) ô: 2.40 (s, 3H, CH3COO), 3.37 (s, N-CH3), 3.39 (s, N-CH3), 3.99 (s)

UV (H20) Xmax: 226 (e 13144)

Analysis: calculated for CnH24N2OSBrJ: C 30.08%, H 5.51%, N 6.38%

found: C 30.48%, H 5.53%, N 6.86%

B. l- (2-Mercaptoethyl) -l, 4,4-trimethylpiperazinium-bis-chloride s-i er

-—V

2a

To a cold (5 ° C) solution of (5R, 6S) -p-nitrobenzyl-6-15 [lR-hydroxyethyl] -3,7-dioxo-l-azabicyclo [3.2.0] heptane-2R-car-boxylate (0.94 g, 2.7 mmol) in 3 ml of acetonitrile (stored under a nitrogen atmosphere), dropwise diisopropylethylamine (0.557 ml, 3.2 mmol) and diphenyl chlorophosphate (0.663 ml, 3.2 mmol) are added, the reaction mixture is stirred for 30 min at 20 5 ° C and treated with diisopropylethylamine (0.599 ml, 3.44 mmol) and an aqueous solution (4 ml) of l- (2-mercap-toethyl) -l, 4,4-trimethylpiperazinium-bischlorid (0.90 g, 3.44 mmol). After 1.25 h, diisopropylethylamine (0.1 ml, 0.57 mmol) is added and the mixture is stirred for a further 2 h. A portion of the acetonitrile 25 is removed under reduced pressure and the red mixture obtained is chromatographed on a PrePak-500 / C18 (Waters Associates) column, using 25-75% acetonitrile in water as the eluent. A yellowish powder (1.4 g) is obtained after lyophilization. The powder is solubilized in water 30 and the solution is placed on a Permutit S-1 Cl "column (1.2x58 cm) using water as the eluent. Lyophilization of the appropriate fractions leads to 1.17 g of a powder which it is again purified on a column of PrePak-500 / C18, the appropriate fractions are lyophilized and a 35 yellowish powder (0.80 g, 53%) is obtained.

IR (KBr) vmax: 3400 (br, OH), 1770 (C = 0 of ß-lactam), 1690 (C = 0 of the PNB ester), 1605 (aromat.), 1515 (N02), 1345 (N02) cm "" 1

MMR (D20) ô: 1.26 (d, J = 6.3 Hz, 3H, CH3CHOH), 3.39 "o (s, NCH3), 4.00 (s), 5.37 (brs, CH2 by PNB) , 7.60 (d, J = 8.6Hz, 2H, Ho from PNB), 8.20 (d, J = 8.7 Hz, 2H, Hm from PNB) UV (H20)). Max: 276 (e 12094), 306 (e 10752)

Analysis: calculated for C25H36N406SC12-3H20: C46.51%, H6.56%, N8.68%, S 4.97%, 45 Cl 10.98%

found: C46.31%, H6.18%, N8.57%, S5.36%,

Cl 11.37%

D. (5R, 6S) -3- [2- (1,4,4-trimethyl-l-piperazinium) -ethylthio] -6-50 (IR) -hydroxyethyl] -7-oxo-l-azabicyclo [3.2. 0] hept-2-en-2-car boxylate chloride

A mixture of 1- (2-acetylthioethyl) -1, 4,4-trimethylpi-perazinium bromide iodide (1.84 g, 4.19 mmol) and 6N hydrochloric acid (15 ml) is heated under a nitrogen atmosphere for 2.5 h 57 ° C. The solution is concentrated to dryness under reduced pressure, the solid is suspended in 10 ml of water and the well-stirred mixture is treated with Permutit S-1 CL "until a solution is obtained. The solution is poured onto a Permutit S-1 CL" - Column (1.2 x 60 cm), which is eluted with water (1.5 ml / min), combines the appropriate fractions and lyophilized, giving a white powder (0.93 g, 85%), mp. 190 up to 191 ° C.

IR (Nujol) vmax: 2460 (SH)

JH-NMR (D20) Ô: 3.4 (s, N-CH3), 3.45 (s, N-CH3), 4.07 (s) Analysis: for C9H, 2N2SClv0.75H2O calculated: C 39.34 %, H 8.62%, N 10.20%, S 11.67% found: C 39.48%, H 8.39%, N 10.55%, S 11.15%

55

60

A mixture of (5R, 6S) -p-nitrobenzyl-3- [2- (l, 4,4-tri-methyl-l-piperazinium) -ethylthio] -6- (lR) -hydroxyethyl] -7-oxo l-azabicyclo [3.2.0] hept-2-en-2-carboxylate-bis-chloride (0.40 g, 0.68 mmol), phosphate buffer (30 ml, 0.05 M, pH 7.0), tetrahy -65 drofuran (10 ml), ether (30 ml) and 10% Pd / C (0.40 g) are hydrogenated for 1 h at 23 ° C. and a pressure of 2.4 bar (35 psi). The two phases are separated, the organic phase is extracted with water (10 ml), the aqueous phase is filtered through a Celite

658 248 2

Pillow, washed with 10 ml ether, concentrated in vacuo to 10 ml and chromatographed on a PrePak 500 / C18 column (2.2x11 cm) (with water as eluent). After lyophilization, 70 mg (25%) are obtained.

IR (KBr) vraax: 3400 (br, OH), 1755 (C = 0 of β-lactam), 1585 (carboxylate) cm "1

^ -NMR (D, 0) ò: 1.24 (3H, d, J = 6.3 Hz, CH3CHOH), 3.36 (s, NCH,), 3.98 (s)

UV (H-.0) 296 (e 7987)

[cc] d = 35.9 ° (C 0.30, H20)

ti2: 9.8 h (measured at a concentration of 10 "4 M in pH 7.4 phosphate buffer at 36.8 ° C).

M

Claims (8)

658 248 2nd PATENT CLAIMS 1. Process for the preparation of carbapenem derivatives of Formula I ' K8 H S-A-R 14 : © COOR 2 ' I " wherein R8 represents a hydrogen atom and R1 represents a hydrogen atom or represents the following substituted and unsubstituted radicals: an alkyl, alkenyl and alkynyl group having 1 to 10 carbon atoms; a cycloalkyl and cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl units; a phenyl group; an aralkyl, aralkenyl and aralkynyl group, wherein the aryl unit represents a phenyl group and the aliphatic part has 1 to 6 carbon atoms; a heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl group, the hetero atom or the hetero atoms in the abovementioned heterocyclic units being selected from 1 to 4 oxygen, nitrogen and sulfur atoms and the alkyl units connected to the heterocyclic units 1 have up to 6 carbon atoms; wherein the substituent or substituents of the above radicals mean one of the following groups; a C 1-4 alkyl group which is optionally substituted by amino, halogen, hydroxy or carboxyl, a halogen atom, A represents a cyclopentylene, cyclohexylene or a C2_6-alkylene group which is optionally substituted by one or more Cj_4-alkyl groups; R2 'represents carboxyl protecting group, 5 X® represents a counter anion, and R14 denotes a quaternized, nitrogen-containing, aromatic or non-aromatic heterocycle which is bonded to A via a ring nitrogen atom, whereby a quaternary ammonium group is formed, or a pharmaceutically acceptable salt thereof, characterized in that a compound of formula IV »8 H 15 COOR 10th IV 20 wherein R1 and R8 have the meanings given above, R2 'represents a carboxyl protective group and L represents a leaving group, with a thiol compound of the formula 25th HS-A-R14 X0 (VII) 30th o o -CR3, -ocnr3r4, -cnr3r4, -nr3r4, No' hr3r4 wherein A, R14 and Xe have the meanings given above, are reacted in an inert solvent and in the presence of a base, and the compound obtained is optionally converted into a pharmaceutically acceptable salt thereof. 2. Process for the preparation of a carbapenem compound of the formula 35 R8 H 0 0 -so2nr3r4, -nh £ nr3r4, r3cnr4-, -cogr3, «o, 0 0 0 -ocr3, -sr3, -§r9, -§r9, -cn, -nj, -osojr3, 8th -OSOpR3, -NR3S09R4, -NR3C = NR4, -NR3C0oR4 or t-x R3 -no2, where in the above substituents the groups R3 and R4 independently represent a hydrogen atom; an alkyl, alkenyl and alkynyl group having 1 to 10 carbon atoms; a cycloalkyl, cycloalkylalkyl and alkylcycloalkyl group having 3 to 6 carbon atoms in the cycloalkyl ring and having 1 to 6 carbon atoms in the alkyl units; a phenyl group; an aralkyl, aralkenyl and aralkynyl group, wherein the aryl unit is a phenyl group and the aliphatic part has 1 to 6 carbon atoms; or a heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl group, the heteroatom or the heteroatoms in the abovementioned heterocyclic units being selected from 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl units connected to the said heterocyclic units 1 to 6 carbon atoms exhibit, stand, or R3 and R4 together with the nitrogen atom to which at least one of these radicals is bonded form a 5- or 6-membered nitrogen-containing heterocyclic ring; R9 is defined as the group R3, but cannot mean a hydrogen atom; or in what R1 and R8 together represent a C2-10 alkylidene radical or a C1-10 alkylidene radical substituted by a hydroxyl group; 40 AT 'S-A-R "COOR2 14 (I ") wherein R1, R8, A and R14 have the meanings given in claim 1 and R2 represents hydrogen or an anionic charge, in the case when R2 "is hydrogen 45 a counter anion X® is also present, or a pharmaceutically acceptable salt thereof, characterized in that, according to the process according to claim 1, a compound of the formula I ', in which R1, R8, R2, A, R14 and X® has the content in claim 1 have the meaning given, produced, and in the 50 NEN compound removed the carboxyl protecting group R2 ', and the compound obtained optionally converted into a pharmaceutically acceptable salt thereof. 3. The method according to claim 1 or 2, characterized in that the base is a non-nucleophilic, tertiary amine 55 or a tri (Cw) alkylamine. 4. The method according to claim 3, characterized in that a non-nucleophilic, tertiary amine is used as the base. 5. The method according to any one of claims 1-4, characterized in that the implementation of the compound of 60 formula IV with the compound of formula VII at a temperature of -15 ° C to room temperature. 6. The method according to claim 5, characterized in that one carries out the reaction at -15 to +15 ° C. 7. The method according to any one of claims 1-6, characterized 65 characterized in that acetonitrile, a mixture of acetonitrile, dimethylformamide and tetrahydrofuran, a mixture of tetrahydrofuran and water, a mixture of acetonitrile and water and acetone are used as the inert solvent. 3rd 658 248 8. Quaternary aminthiol compounds of the formula HS-A (R14 X® wherein A is cyclopentylene, cyclohexylene or (VII) , 10th l11 R12 I C - ^ 13 is in which R10, R11, R12 and R13 independently of one another represent a hydrogen atom or a CM-alkyl group, X® represents a counter anion and R14 represents a substituted or unsubstituted, mono-, bi- or polycyclic, aromatic or non-aromatic, heterocyclic Represents radical which contains at least one nitrogen atom in the ring and is bonded to A via a ring nitrogen atom, whereby a quaternary ammonium group is formed, as a means for carrying out the process according to claim 1 or 2. unit represents a phenyl group and the aliphatic part has 1 to 6 carbon atoms; a heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl group, the hetero atom or the hetero atoms in the abovementioned heterocyclic units being selected from 1 to 4 oxygen, nitrogen and sulfur atoms and the alkyl units connected to the heterocyclic units Have 1 to 6 carbon atoms; wherein the substituent or the substituents of the abovementioned radicals mean one of the following groups: a C 1-4 alkyl group which is optionally substituted by amino, halogen, hydroxy or carboxyl, a halogen atom, 15