LU85241A1 - PROCESS FOR THE PREPARATION OF CARBAPENEM DERIVATIVES, NOVEL SYNTHESIS INTERMEDIATES AND PROCESS FOR THE PREPARATION OF SUCH INTERMEDIATES - Google Patents

Description

D.52.216

Patent application of March 7, 1984 Designation of the Inventor (1) The undersigned .. Jacques de Muyser ............................. ......................................... ......... .................................................. ......................

.................................................. ...... 35d, _b ^ ulevar (â .__ 3 ^ Qyja.l ..... à .... LüXEMB.QnEG .................. .................................................. ............

i acting as desdepasantr- agent of the depositor - (2) .la ..... company called: BRXSTQIgrMYERS-, CQMPANY ...................... .................................................. ...........................

345 Park Avenue NEW YORK, NYlol54 (United States of America) (3) of the intention concerning: ...... "Process ... ... preparation ..... of ... carbap derivatives. ênem, .. no.uveaux .... inter: = synthesis media and preparation process for these ..... i.nb.er.-.

"designates as inventor (s): 1. Surname and first names ..REXTRAZE ................................ .............! .................................... .................................................. .................

Address 6600 Place de la Bataille, LAPRAIRIE, PQ (Canada) 2. Last and first names ................................ .................................................. ............................................. ..... .................................................

Address ................................................. .................................................. .................................................. .................................................. .........................................

3. Surname and first names ............................................. .................................................. .................................................. .................................................. ..........

Address ................................................. ............................! ..................... ................ .................................. .................................................. ..............................

He affirms the sincerity of the above indications and declares to assume full responsibility for them.

.............. Luxembourg f ie ................................. ............................................... 19 84 { Ækm (signature) (1) Last name, first names, company, address.

(2) Last name, first names and address of the depositor.

(8) Title of the invention as in the patent application. T. s. v. p.

OC 52 21b

PRIORITY RENDERING

of the patent application / utility model Εκ IN THE UNITED STATES OF AMERICA FROM 8 MARCH 1983 17 JUNE 1983_ 1 DECEMBER 1983 Brief Description filed in support of a request for

PATENT

at

Luxembourg

in the name of: BRISTOL-MYERS COMPANY

for: "Process for the preparation of carbapenem derivatives, new synthesis intermediates and process for the preparation of these intermediates".

The present invention relates to a new process for the preparation of carbapenem derivatives, new synthesis intermediates and a process for the preparation of these intermediates. More particularly, the present invention relates to a new process for the preparation of carbapenem derivatives having a substituent in position 2 of the formula -SA-R14 in which A represents an optionally substituted cyclopentylene, cyclohexylene or 10o-C 4 alkylene radical. at 14

by one or more alkyl groups in and R

represents an aromatic or non-aromatic heterocycle containing a nitrogen atom in quaternary form linked to A by a quaternary nitrogen atom.

The carbapenem derivatives prepared by the process of the present invention are disclosed and claimed by Messrs. Choung U. Kim and Peter F. Misco, Jr, in American patent applications filed under Nos. 366,910, 471,379 and 389,652 respectively. April 9, 1982, March 8, 1983 and June 18, 1982; whose entire content of each of these requests is incorporated herein by reference.

US application 366,910 and its application for partial continuation filed under No. 471,379 of March 8, 1983, which request for partial continuation 25 corresponds to the patent application for Federal Germany published under No. 3,312,533, reveal the preparation of carbapenem antibiotics of the formula ci 2 o HE 1 R1--

5 N-L

X,

COOR

IA

8 1: ¾ 10 in which R is hydrogen and R is chosen from the group consisting of hydrogen; the following substituted and unsubstituted groups: alkyl, alkenyl and alkynyl, having from 1 to 10 carbon atoms; cycloalkyl and cycloalkylalkyl, having from 3 to 6 carbon atoms in the cycloalkyl ring and from 1 to 6 carbon atoms in the alkyl parts; phenyl; aralkyl, aralkenyl, and aralkynyl in which the aryl part is phenyl and the aliphatic part has from 1 to 6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and hetero-cyclylalkyl in which the hetero atom (s) in the heterocyclic moieties mentioned above are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and alkyl parts associated with said heterocyclic parts have from 1 to 6 carbon atoms; in which the substituent or the substituents with respect to the radicals mentioned above are chosen from the group consisting of: C 1 -C 6 alkyl group optionally substituted by amino, halogen, hydroxy or halogenated carboxyl 3 -0R ^ 0

It 3 4

-OCNR ^ R

3

8 3 4 -CNÏTFT

3 4

-mJR

5 / NR3 -f 'NR3R4 10 -S02NR3R4

S 34 -NHCNR ^ IT

0 • 5II 4 R ^ CNR - 15 -co2r3 = 0 fi3 20 -OCR "5 -SR3 8 9 -SRy 25. 0 -Ir9 11 0

-CN

30 -N3 -OSO ^ R3 -oso2r3 35 -NR3S02R4 4 -nr3c = nit b 3 4

-Nirco2R

5 -N02 in which, with respect to the substituents mentioned 3 4 above, the groups R and R are independently chosen from hydrogen; an alkyl, alkenyl and alkynyl group, having from 1 to 10 carbon atoms, cycloalkyl, cycloalkyl-alkyl and alkylcycloalkyl, having from 3 to 6 carbon atoms in the cycloalkyl ring and from 1 to 6 carbon atoms in the alkyl parts ; phenyl; aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has from 1 to 6 carbon atoms; and heteroaryl, hereroaralkyl, heterocyclyl and heterocyclylalkyl in which the hetero atom or atoms in the heterocyclic moieties mentioned above are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the moieties alkyl associated with said heterocyclic parts have from 1 to 6 carbon atoms, or R and R taken together with the nitrogen to which at least one is linked or attached may form a heterocyclic ring containing nitrogen with 5 or 6 members ; R is as defined for R except that it cannot be hydrogen; or in which R and R taken together represent a C2-C10 alkylidene group; U alkylidene substituted by hydroxy; A is cyclopentylene, cyclohexylene or C 1 -C 6 alkylene optionally substituted by one or more C 1 -C 4 alkyl groups; R is hydrogen, an anionic charge or a conventional easily separable carboxyl protecting group, provided that when R is hydrogen or a protecting group, a counter anion is also present; and © / "-o represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocyclic radical containing at least one nitrogen in the nucleus and linked to A by a nitrogen in the nucleus, thus forming a quaternary ammonium group; and the pharmaceutically acceptable salts thereof by the process shown in the following reaction scheme: 10

Be I

0.01 diphenyl chlorophosphate N ^ ^ COOR2 ’

III

20 O H 0 R8 - It

r14_1 / \ ^ op (oc6h5) 2 HS-A-OH

I l - J — N- 25 0 COOR ^ A = C ^ -Cg alkylene or cycloalkylene

30 _8 H

1 ^ S-A-OH ^ - I chloride methanesulfonyl I | - Â N- 0 COOR ^ 1

V

35 6

8 a.m.

, Ξ / \. S-A-OSO CH ΙΘ R - 1 T - * X-N-U. 2 ’5 0 ^ ^ COOk

VI

10 4rr ‘" Q

C (X N C00R2 'Αζ> ^ (XV =. Against anion)

15 II

"8 H Θ f \

20 K Ξ yv / S-A-N J

R1 - T Π W

l -►-

A-N-V

q / COOR

25 I, A

R8 S βΓ ~.

R'-f + ^ v ‘MJ

30 I

// - N- \ 2 V COOR ^

f IA

7

Application 389,652 and its application for partial continuation 499,690 filed on June 7, 1983 disclose the preparation of carbapenem antibiotics of the formula: 5

R8? r5 / '"“' N

R1-- | ^ VS ~ A \ ^)

i — N —- L

'10 0 ^ COOR2

f IB

o 1 in which R is hydrogen and R is chosen from the group consisting of hydrogen; the following substituted and unsubstituted groups; alkyl, alkenyl and alkynyl, having from 1 to 10 carbon atoms; cycloalkyl and cycloalkylalkyl, having from 3 to 6 carbon atoms in the cycloalkyl nucleus and from 1 to 6 carbon atoms in the alkyl parts; phenyl; aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has from 1 to 6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and -heterocyclylalkyl in which the hetero atom (s) in the heterocyclic moieties mentioned above are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl parts associated with said heterocyclic parts have from 1 to 6 carbon atoms; in which the substituent or the substituents with respect to the radicals mentioned above are independently chosen from the group consisting of: C 1 -C 6 alkyl group optionally substituted by amino, halogen, hydroxy or halogenated carbonyl 35 -OR5 8 fi 3 4 -OCNR ^ ït fi 3 4 -CNR-Tl 5 34 -NR ^ R -, NR3 - ^ 3 4

, \ NR R

-SO2NR3R4; ? -NHCNrV * 3ff 4 R ^ CNR ^ 15 -co2r3 = 0 »3 20 -OCR" * -SR3? 9 -SRy 25 S9 -SR "3

II

ο

-CN

30 -N

3 -OSO ^ R3 3 -0S02R3 9 -NR3S02R4 -nr3ç = nr4 i 3 5 R3 -NR3C02R4 -no2 10 in which, with respect to the substituents mentioned above, the groups R3 and R4 are independently chosen from hydrogen; an alkyl, alkenyl and alkynyl group having from 1 to 10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl, having 3-6 carbon atoms in the cycloalkyl nucleus and 1-6 carbon atoms in the alkyl parts; phenyl; aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has from 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the hetero atom (s) in the heterocyclic moieties mentioned above are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with said heterocyclic moieties have from 3 4 25 1-6 carbon atoms, or R and R taken together with the nitrogen to which at least one is linked may form a heterocyclic ring containing nitrogen with 5 or 6 members; o 3 R is as defined for R with the exception that it cannot

Λ R

not be hydrogen; or wherein R and R taken together represent a C 2 -C 12 alkyl or C 2 -C 6 alkylidene group substituted by hydroxy; R ^ is selected from the group consisting of the following substituted and unsubstituted groups: alkyl, alkenyl and t alkynyl, having 1-10 carbon atoms; cycloalkyl and cycloalkylalkyl, having 3-6 carbon atoms in the cycloalkyl nucleus and 1-6 carbon atoms in the alkyl parts; phenyl; aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has from 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and hetero-5-cyclylalkyl in which the hetero atom or atoms in the heterocyclic parts mentioned above are chosen from the groups consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl parts associated with the heterocyclic parts have from 1-6 carbon atoms; in which the radicals R3 mentioned above; above are optionally substituted with 1-3 substituents independently selected from: a C 1-6 alkyl group optionally substituted with amino, fluoro, chloro, carboxyl, hydroxy or carbamoyl; fluoro, chloro or bromo -OR3; -0C02R3; 20 3 -OCOR ^; -0C0NR3R4; 25. '-0S02R3; -oxo; -NR ^; 30 3 4 R ^ CONR -; -NR3C02R ^; -NR3C0NR3R4; 11 -NR5SO2R4; -SR3; 0 5 -S-r9% / P9 -S-Ry; -SO, R3 10 3 “C02r3> -C0NR3R4; 15 -CN; or phenyl optionally substituted with 1-3 fluoro,

X

chloro, bromo, C ^ -Cg alkyl, -OR, 20 -NR% 4, -SO, R3, -C0 ~ R3 or -C0NR3R4, in X 3 L ^ 5 which R ^, R and R in said substituents R are as previously defined; or R3 may represent a divalent phenylene radical or a divalent C 1 -C 4 alkylene group linked to the ring: Ό 30 so as to form a bridged polycyclic group; A is a cyclopentylene radical, or cyclohexylene or alkylene optionally substituted by one or more C1-C6 alkyl groups; R2 is hydrogen, an anionic charge 35 or a conventional easily removable carboxyl protecting group 2 12, provided that when R is hydrogen or a protecting group, a counter anion is also present; and

5 O

represents a substituted or unsubstituted mono-, bi- or polycyclic non-aromatic heterocyclic radical containing at least one nitrogen in the nucleus and linked to A by a nitrogen in the nucleus, thus forming a quaternary ammonium group; and the pharmaceutically acceptable salts thereof, by the method shown in the following reaction scheme 15 R - (chl orophosphate 'of diphenyl 20 À-N-L „---—- ► CT ^ COOR2'

III

25 R8 I 9

R1__HS-A-OH

l -► 3 ° A-N - \ A = alkylene or 0 ^ C00R2 'C ^ -Cg cycloalkylene

! V

13 R8 ξ pi __ ^ ^ ^ I chloride | methanesulfonyl <y N ~ T! 00R2 ’

5 V

R8 9 '* ^ S-A-0S0oCHx R - T if ιθ 10 ✓? - N-L 21 * "^ COOR ^

VI

15 rU- ^ j ^ 3 - * - 1 <0 yy N \ 2 ’Ae® X® <f COOR g

II

20 R8 1 # ^ 1 __ ^ ^ release I possible —- *> / y N \ 21 0X COOR ^

25 I’B

n8 H Κ / ^ ~ Λ 3o "4tr ° J-N 2 COOR ^

- IB

14

To work on the scheme of the prior art, the starting material of formula III is reacted in an inert organic solvent with diphenyl chlorophosphate in the presence of a base to give intermediate 5 of formula IV. Intermediate IV is then reacted with a mercaptan reagent of the formula HS-A-OH in an inert organic solvent and in the presence of a base to give an intermediate of formula V. Intermediate V is then acylated with methanesulfonyl chloride in an inert organic solvent and in the presence of a base to give the intermediate of formula VI which is reacted with a source of iodide ions in an inert organic solvent to give the intermediate of formula II. Intermediate II is reacted with the desired amine in an inert organic solvent and in the presence of a silver ion to produce the quaternary product I'A or I'B which can then be unlocked to give the unlocked carbapenem correspondent of formula IA or IB.

The process described above has several disadvantages. Thus, for example, the method includes several steps which could advantageously be reduced in number. The total reaction yield is also quite low and the stage of transformation into a quaternary ion is carried out on the complete carbapenem compound. It would be desirable to carry out a new process for the production of compounds of formula IA or IB which (1) includes fewer reaction steps, (2) gives higher yields, (3) makes it possible to form the quaternary amine in first and then to link the carbapenem nucleus in a later step in the synthesis and (4) can be used to more easily form quaternary amine products with ", a large variety of amines, ie amines with steric hindrance and those with low pK ^ values.

15

The present invention makes it possible to meet all these technical requirements for the first time. Thus, the present invention provides a new process for the preparation of carbapenem derivatives of the formula 5 R8 1 R1 __ = ^ \ ^ SA-R14 10 J> -N- \ cr COOn 8 i 15 in which R is hydrogen and R is selected from the group consisting of hydrogen; the following substituted and unsubstituted groups: alkyl, alkenyl and alkynyl, having 1-10 carbon atoms; cycloalkyl and cycloalkylalkyl, having 3-6 carbon atoms in the cycloalkyl nucleus and 1-6 carbon atoms in the alkyl parts; phenyl; aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has from 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the hetero atom (s) in the heterocyclic moieties mentioned above are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with said heterocyclic portions have 1-6 carbon atoms; In which the substituent or substituents with respect to the radicals mentioned above are chosen from the group consisting of: C 1 -C 6 alkyl group optionally substituted with amino, halo, hydroxy or carboxyl.

halogen -OR3 16

-oI! nr3r4 _ ff ^ L

5 -CNR ^ R ^ -nr3r4 .NR3 10 34 -so2nr3r4 -nhc! Nr3r4 15 J 4 R ^ CNR - -co2r3 = 0 20 »3 -OCR5 -SR3

P

25 »9 -SRy

B

-CN

50 -0S03R3 -0S0oR3 35 2 17 3 _ Lî -nrjso2r -nr3c = nr4 l3 5 R · 5 -nr3co2r4 -no2 10 in which, compared to the substituents mentioned above, 34, the groups R and R are independently chosen from hydrogen; an alkyl, alkenyl and alkynyl group having 1-10 carbon atoms; cycloalkyl, cycloalkyl-alkyl and alkylcycloalkyl, having 3-6 carbon atoms in the cycloalkyl nucleus and 1-6 carbon atoms in the alkyl parts; phenyl; aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and hetero-cyclylalkyl in which the hetero atom (s) in the heterocyclic moieties mentioned above are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with the heterocyclic moieties have 1-6 carbon atoms, or R and R taken together with the nitrogen to which at least one is bonded can form a heterocyclic ring containing nitrogen at 5 or 6 members; R ^ is such that • 3 defined for R except that it cannot be hydrogen; 1 8 or in which R and R taken together represent a

30 ^ “^ lO alkyl radical or ^“ ^ lO alkyl radical

substituted by hydroxy; A is a cyclopentylene, cyclohexylene or C 2 -C 6 alkylene radical optionally substituted by one or more C 1 -C 4 alkyl groups; R ^ is hydrogen, an anionic filler or a conventional easily removable carboxyl protecting group, ^ 0 provided that when R is hydrogen or a protecting group 18, a counter anion is also present; 14 and R is an aromatic or non-aromatic heterocycle containing nitrogen in quaternary form linked to A by nitrogen in the nucleus, thereby forming a quaternary ammonium group, or a pharmaceutically acceptable salt thereof, said process being characterized in that it includes the reaction of the intermediate of the formula:

Oh

10 R ° - N T00R2 '

15 IV

1 8 in which R and R are as defined above, 2 ′ R is a conventional easily detachable carboxyl protecting group and L is a conventional detachable group, with a thiol compound of the formula: HS-A-R14 X® VII - 14 in which A and R are as defined above and X "is a counter anion, in an inert solvent and in the presence of a base, to produce a carbapenem product of the formula: 19 R8?

Ri l> ys · * -8 ΠΙ ^ 5 f N COOR2 1 8 2 '14 in which R, R, R, A, R and X are as defined above and, if desired, removal of the group 2' 10 of carboxyl protection R to give the corresponding unlocked compound of formula I or a pharmaceutically acceptable salt thereof.

The present invention also relates to novel synthesis intermediates of formula VII and methods for the preparation of such intermediates.

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

The above-mentioned compounds of general formula I contain the carbapenem ring: £ - 11_N-13 25 0 4 and can therefore be called as derivatives of 1-carba-2-penem-3-carboxylic acid. Alternatively, the compounds may be considered to have the basic structure 20 4 & - J * -, 2 5 and be termed as 7-oxo-1-azabicyclo (3.2.0) hept-2- acid derivatives ene-2-carboxylic. While the present invention includes compounds in which the relative stereochemistry of the protons 5,6 is cis as well as trans, the preferred compounds have the stereochemistry 5R, 6S (trans) as in the case of thienamycin.

The compounds of formula I can be unsubstituted in position 6 or substituted with. substitution groups previously described for other derivatives

O

15 carbapenem. More specifically, R can be hydrogen 1 and R can be hydrogen or a substituent different from hydrogen described, for example, in European patent application 38 869 (see the definition of R / -).

Alternatively, R and R taken together can be a C 2 -C 10 alkylidene or C 1 -C 4 alkylene substituted, for example by hydroxy.

1 8

To work on the definitions of R and R: (a) The aliphatic "alkenyl", "alkenyl" and "alkynyl" groups may be straight or branched chain having 1-10 carbon atoms; groups of 1 are preferred -6, more preferably 1-4, carbon; when part of another substituent, for example as in the cycloalkylalkyl, or heteroaralkyl or aralkenyl group, the alkyl, alkenyl and alkynyl group preferably contains 1-6 , more preferably 1-4, carbon atoms.

(b) "heteroaryl" includes mono-, bi- and polycyclic aromatic heterocyclic groups containing 1-4 atoms of 0, N or S; preferred are heterocyclic rings 21 with 5 or 6 members such as thienyl, furyl, thiadiazolyle, oxadiazolyle, triazolyle, isothiazolyle, thiazolyle, imidazolyle, isoxazolyle, tetrazolyle, oxazolyle, pyridyle, pyrazinyle, pyrimidinyle, pyridazyn, pyridazin

(c) "heterocyclyl" includes mono-, bi- and polycyclic saturated or unsaturated non-aromatic heterocyclic groups containing 1-4 atoms of 0, N or S; i prefer heterocyclic rings with 5 or 6 members such as morpholinyl, piperazinyl, piperidyl, pyrazoli-dinyl, imidazolinyl, imidazolidinyl, pyrrolinyl, pyrrolidinyl, etc.

(d) "halogen" includes chlorine, bromine, fluorine and iodine and is preferably chlorine or bromine.

The term "conventional easily detachable carboxyl protecting group" refers to a known ester group which has been used to block a carboxyl group during the chemical reaction steps described below and which can be removed, if desired, by methods Which do not result in any appreciable destruction of the remaining part of the molecule, for example by chemical or enzymatic hydrolysis, treatment with chemical reducing agents under mild conditions, irradiation with ultraviolet light or catalytic hydrogenation. Examples of such ester protecting groups include benzhydryl, p-nitrobenzyl, 2-naphthylmethyl, allyl, benzyl, trichloroethyl, silyl such as trimethylsilyl, phenacyl, p-methoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl and alkyl ^ -Cg such as methyl, ethyl or butyl. Also included in such protective groups are those which are hydrolyzed under physiological conditions such as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethyl.

Particularly advantageous carboxyl protecting groups are p-nitrobenzyl groups which can be easily removed by catalytic hydrogenolysis and allyl which can be removed by reaction catalyzed by Pd (P0j) 4.

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

Compounds of formula I in the form of acid addition salts can be written as R8 5 R-TI ^ 20 c / ~ N \ x) OR2 p R: H or a protecting group 25 in which X represents the acid anion. The counter anion X ~ can be chosen so as to provide pharmaceutically acceptable salts for therapeutic administration but, in the case of the intermediate compounds of formula I, X “can also be a toxic anion.

In such a case the ion can be subsequently removed or substituted with a pharmaceutically acceptable anion to form an active final product for therapeutic use.

When acidic or basic groups are present in the R group or on the radical R in quaternary form, the present invention may also include suitable base or acid salts of these functional groups, for example salts of addition of acid in the case of a basic group and metal salts (e.g. sodium, potassium, calcium and aluminum), ammonium salt and salts with non-toxic amines (e.g. trialcoylamines, procaine , dibenzylamine, 1-ephenamine, N-benzyl- ^ -phenethylamine, Ν, Ν'-dibenzylethylenediamine, etc.) in the case of an acid group.

2-5 Compounds of the formula I in which R is hydrogen, an anionic charge or a physiologically hydrolyzable ester group as well as pharmaceutically acceptable salts thereof are useful as antibacterial agents. The remaining compounds of formula I are valuable intermediates which can be converted into the biologically active compounds mentioned above.

A preferred embodiment of the present invention comprises compounds of formula I wherein 8 1 R is hydrogen and R is hydrogen,

CH, CH, OH OH

^ l Y

^ pCH-, or CH ^ H- ch3 ch3 25

Among this subclass, the preferred compounds are those

AT

in which R is

OH

CH3CH-, the more preferred compounds having the absolute configuration 5R, 6S, 8R.

Another preferred embodiment comprises compounds of formula I in which R and R taken together form an alkylidene radical of the formula: 24 H0CH2 c = / CH * 5 3

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

10 cyclopentylene H ^ C HC

£ H2C-CH ^ 15 / / HC / \ cyclohexylene / H2C HC \

H2C ^ C- / CH2 J

V H z J

20 ^ κ or a C2-C6 alkylene radical optionally substituted by one or more C ^ -C ^ alkyl substituents.

Preferred substituents A are the cyclopentylene, cyclohexylene or alkylene radical of the formula: 25 R10 r12 I i -c-c- l11 b ΊΟ ΊΊ Ίp · ι? Wherein R, R, R and Ru are each independently hydrogen or a C 1 -C 4 alkyl group. A preferred embodiment comprises the compounds of formula I in which the substituent A is -CH2CH2 ~, -CHCH2 ~,

<L

5 25 - (\ or -CH0CH- yv 2i / CH3

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

14

The substituent R in quaternary form can be an optionally substituted mono-, bi- or polycyclic aromatic or non-aromatic heterocyclic radical containing at least one nitrogen in the nucleus and linked to A by a nitrogen in the nucleus, thus forming a quaternary ammonium group .

14

A preferred class of substituents R can be represented by the general formula:

20 -N J

which is intended to define a substituted or unsubstituted mono-, bi- or polycyclic heteroaryl radical containing at least one nitrogen in the nucleus and linked to a carbon atom of the substituent A by a nitrogen in the nucleus, thus forming an ammonium group quaternary. The heteroaryl radical may be optionally substituted by substituents such as C 1 -C 4 alkyl groups, alkyl substituted by hydroxy, amino, carboxy or halogen, C 1 -C 6 cycloalkyl> C 1 -C 6 alkylthio ^, amino, C 1-6 alkylamino | di (C ^ -C ^ alkyl) amino, halogen, C ^ -C ^ alkanoylamino, C ^ -C ^ alkanoyloxy, carboxy, 0 -b-0C1-C4 alkyl, 26 hydroxy, amidino, guanidino, trifluoromethyl, phenyl, phenyl substituted by one, two or three amino groups, halogen, hydroxyl, trifluoromethyl, G ^ -C ^ alkyl or C ^ -C ^ alkoxy, heteroaryl and heteroalkyl in which the hetero atom or atoms in the heterocyclic parts mentioned above are chosen from the group consisting of 1-4 atoms of 0, N or S and the alkyl part associated with the heteroaralkyl a of 1-6 atoms of carbon.

>! 0 The heteroaryl radical linked to the substituent A is preferably an aromatic heterocyclic radical with 5 or * 6 members containing a nitrogen atom in quaternary form (which is directly linked to the carbon atom of the alkylene or cycloalkylene radical) and , optionally, one or more additional hetero atoms chosen from 0, N or S. While, in general, any heteroaryl radical linked to A via a nitrogen atom in quaternary form has been found to produce biologically active carbapenem derivatives, a preferred embodiment comprises compounds in which -Ό 25 represents a radical chosen from the group consisting of: (a) 30 R5 "c ÇL rj in which R, R ° and R 'are independently chosen from hydrogen; C 1-6 alkyl, C 55 substituted by hydroxy, amino, carboxy or halogen; 27 C 1-6 cycloalkyl; C alkoxy; C 1-6 alkylthio; amino, C 1-6 alkylamino ^; di (C ^ -C ^ alkyl) - amino; halogen; C ^ -C ^ alkanoylamino; CrC4 alkanoyloxy; carboxy; 5 8 -C-OC ^ -C ^ alkyl; hydroxy, amidino; guanidino; trifluoromethyl; phenyl; phenyl substituted with one, two or three amino, halogen, hydroxyl, trifluoromethyl, C 1 -C 4 alkyl or C 1 -C 4 alkoxy; and heteroaryl and heteroaralkyl in which the hetero atom or atoms in the heterocyclic parts mentioned above are chosen from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part associated with said heteroaralkyl part has 1-6 carbon atoms; 5 6 7 15 or wherein two of R5, R ° or R 'taken together may be a fused saturated carbocyclic ring, a condensed aromatic carbocyclic ring, a condensed saturated heterocyclic ring or a condensed heteroaromatic ring, 20

(b) 5 ^ ®N

Π fl fl] 25 W ’or optionally substituted on a carbon atom by one or more substituents independently chosen from C 1 -C 4 alkyl groups; C 1 -C 4 alkyl substituted by hydroxy, amino, carboxy or halogen; C ^ -C6 cycloalkyl C <j-C4 alkoxy; alkylthio en; amino; C 1 -C 4 alkylamino di (C 1 -C 4 alkyl) amino; halogen; C ^ -C ^ alkanoylamino; C1-4 alkanoyloxy carboxy; 8 35 —C — 0C> | —C4 alkyl; hydroxy; amidino; guanidino; , ---- 28 trifluoromethyl; phenyl; phenyl substituted with one, two or three amino, halogen, hydroxyl, trifluoromethyl, C 1 -C 4 alkyl or C 1 -C 4 alkoxy group; and heteroaryl or heteroaralkyl in which the hetero atom or atoms in the heterocyclic parts mentioned above are selected from the groups consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part associated with said heteroaralkyl part has 1-6 carbon atoms, or optionally substituted so as to form a condensed carbocyclic or heterocyclic ring; © I gJ © j (c) 15

»F '·', -ys A

U U "U

Optionally substituted on a carbon atom by one or more substituents independently selected from C 1 -C 4 alkyl groups; C 1 -C 4 alkyl substituted by hydroxy, amino, carboxy or halogen; C ^ -Cg cycloalkyl; C 1 -C 4 alkoxy; C 1 -C 4 alkylthio; amino; C 1 -C 4 alkylamino; di (C 1 -C 4 alkyl) amino; halogen; C 1 alkanoyl amino] C 1; C 1-6 alkanoyloxy; carboxy; 0

II

-C-OC ^ -C ^ alkyl; hydroxy; amidino; guanidino; trifluoromethyl; phenyl; phenyl substituted with one, two or three amino, halogen, hydroxyl, trifluoromethyl, C 1-4 alkyl or C 1-4 alkoxy groups; and heteroaryl or heteroaralkyl in which the hetero atom or atoms in the heterocyclic parts mentioned above are chosen from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part associated with said heteroaralkyl part has 1-6 carbon atoms, or optionally substituted so as to form a condensed carbocyclic or heterocyclic ring; , «> Ày J'y U u u ·

15 I

J © © I

YEAR

K r M tr n

I 11, I fl or I II

20 N ^ N-N N ^ N

optionally substituted on a carbon atom by one or more substituents independently chosen from C 1 -C 4 alkyl groups; C 1 -C 4 alkyl substituted by hydroxy, amino, carboxy or halogen groups; C 1 -C 4 alkoxy cycloalkyl; C 1 -C 4 alkylthio; amino; C 1 -C 4 alkylamino; di (C 1 -C 4 alkyl) amino; halogen; C ^ -C ^ alkanoylamino; C 1 -C 4 alkanoyloxy; carboxy; 3 ° j —C — 0C> j —alkyl; hydroxy; amidino; guanidino; trifluoromethyl; phenyl; phenyl substituted by one, two or three t amino, halogen, hydroxyl, trifluoromethyl, C 1 -C 4 alkyl or alkoxy group; and heteroaryl or heteroaralkyl in which the hetero atom (s) in the heterocyclic moieties mentioned above are selected from the groups consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moiety associated with said heteroaralkyl part has 1-6 carbon atoms, or optionally substituted so as to form a condensed carbocyclic or heterocyclic ring; (e) O © O »0

10 X

in which X is 0, S or NR in which R is a C 1 -C 4 alkyl or phenyl group, said radical being optionally substituted on a carbon atom by one or more substituents independently chosen from C 1 alkyl groups -C ^; C 1 -C 4 alkyl substituted by hydroxy, amino, carboxy or halogen; C ^ -C ^ cycloalkyl; C 1 -C 4 alkoxy, C 1 -C 4 alkylthio; amino; C 1 -C 4 alkylamino; di (C 1 -C 4 alkyl) amino; halogen; C 1-6 alkanoylamino; C 1 -C 4 alkanoyloxy; carboxy; 8 -C-OC ^ -C ^ alkyl; hydroxy; amidino, guanidino; trifluoromethyl; phenyl; phenyl substituted with one, two or three amino, halogen, hydroxyl, trifluoromethyl, C1-C6 alkyl or C1-C6 alkoxy groups; and heteroaryl or heteroaralkyl in which the hetero atom or atoms in the heterocyclic parts mentioned above are chosen from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part associated with 30 said heteroaralkyl part has from 1-6 carbon atoms, or optionally substituted so as to form a ring: condensed carbocyclic or heterocyclic; "31" -Ο. Ô 5 © Θ X — N- N —- N- C 5 ° u L 5 1 ° (r in which X is 0, S or NR in which R is an alkyl or phenyl group, said radical optionally substituted on a carbon atom by one or more substituents independently chosen from C 1 -C 4 alkyl groups; C 1 -C 4 alkyl substituted by hydroxy, amino, carboxy or halogen; C 1 -C 6 cycloalkyl ; C ^ -C ^ alkoxy; C ^ -C ^ alkylthio; amino; C ^ -C ^ alkylamino; di (C ^ -C ^ alkyl) amino; halogen; 20 alkanoylamino; C ^ alkanoyloxy - C4; carboxy; fl -C-OC ^ -C ^ alkyl; hydroxy; amidino; guanidino; trifluoro-methyl; phenyl; phenyl substituted by one, two or three amino, halogen, hydroxyl, trifluoromethyl, C ^ alkyl group - C ^ or C ^ -C ^ alkoxy; and heteroaryl or heteroaralkyl in which the hetero atom or atoms in the heterocyclic parts mentioned above are chosen from the group consisting of 1-4 oxygen and nitrogen atoms or sulfur and the part alkyl associated with said heteroaralkyl portion has 1-6 carbon atoms; and 32 Θ. Θ (g). "=" Γ / "= \

Ik N-R ’Nc ^^ îJ-R

5

N — N îj N-R

® / \, // V

-VR nn ^ S "10 x®

-N-R N N-R

1 / N or 3 / χΛ 15 in which R is a C 1 -C 4 alkyl or phenyl group, said radical being optionally substituted on the carbon atom by a substituent chosen from C 1 -C 4 alkyl groups ; hydroxy substituted C 1 -C 4 alkyl; amino, carboxy or halogen; cycloalkyl; C 1 -C 4 alkoxy; C 1 -C 4 alkylthio; amino; C 1 -C 4 alkylamino; di (C 3 -C 4 alkyl) amino; C1_C4 alkanoylamino "carboxy; 25 g -C-OC ^ -C ^ alkyl; hydroxy; amidino; guanidino; trifluoromethyl; phenyl, phenyl substituted with one, two or three amino groups, halogen, hydroxyl, trifluoromethyl, C 1 -C 4 alkyl or C 1 -C 4 alkoxy; and heteroaryl or heteroaralkyl in which the hetero atom or atoms in the heterocyclic parts mentioned above are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part associated with said heteroaralkyl part has 33 1-6 carbon atoms.

In the above subclass, preferred compounds are those in which the substituent A is 5 -CH £ CH2-, -CHCH2-, - / \ OR -CH2ÇH-; y— * ch3 1 8 and in which either (a) R and R taken together represent 10 hoch2 c £ 3 8 1 15 or (b) R is hydrogen and R represents hydrogen,

CH, ÇH, nH OH

^ X? H! CH3CH2-, '> CH-, or CH3CH ~ Ζΰζ ch3 20

O

Particularly preferred are the compounds in which R

Λ is hydrogen and R is

OH

CH ^ CH-, preferably the compounds having the absolute configuration 5R, 6S, 8R.

A particularly preferred embodiment of the present invention comprises the preparation of the compounds in which: 30 _ + KC3 represents a radical of the formula 5 34 / R6 "NI 7 • 3 6 7 in which R, R and R 'are independently chosen s from the group consisting of hydrogen, of a C 1 -C 4 alkyl, C 1 alkoxy C 1 -C 4 alkyl group. substituted with hydroxy, alkyl, amino, carboxy and carbamoyl. In this subclass, the preferred compounds are those in which the substituent A is -CH2CH2-, -CHCH2-, - / ^ or -CH2CH - ch3 / * ch3

1 O

and in which either (a) R and R taken together represent 20 H0CHo CH ^ or (b) R is hydrogen and R represents hydrogen, 25

CH-, CH-, OH OH

not? ^ i i CH, CH -,> CH-, ^ C- or CH, CH- CÎL CH, 3 3 8

Particularly preferred are those compounds in which R

η is hydrogen and R is

OH

CH ^ CH-, preferably the compounds having the absolute configuration 5R, 6S, 8R.

35

A more preferred embodiment of the present invention comprises the preparation of the compounds in which: 5 -o represents a radical of the formula / r6 10 © 5 6 7 15 in which R, R and R 'are independently chosen from the group consisting hydrogen, C 1 -C 4 alkyl, C 1 -C 4 alkoxy> C 1 -C 4 alkyl substituted by a hydroxy group, C 1 -C 4 alkylthio and amino. In this subclass, the preferred compounds are those in which 2 ° the substituent A is -C ^ CHg -, -CHO ^ -j \ or -CH ^ CH- ch3 ^ 'ch3

<Q

and in which either (a) R and R taken together represent 25 H0CHo CH, 5. . 8 1 30 either (b; R is hydrogen and R represents hydrogen,

CH, CH, OH OH

^ ^ I! CH3CH2 ~ ’^^ CH-, or CH3CH- chT CH, 3 3 36 Ο

Particularly preferred are the compounds in which R

OH

is hydrogen and R ** is CH ^ H-, preferably the compounds having the absolute configuration 5R, 6S, 8R.

Another preferred embodiment of the present invention comprises the preparation of the compounds in which

O

s 10 represents a radical of the formula + —H · —SCHg 15 ^ 'CH3

In this subclass, the preferred compounds are those in which the substituent A is -CHCHg-, -CH ^ C ^ -, 20 --- iH3 - (> or -CH9CH- y— / ch3 Λ 8 and in which either (a) R and R taken together represent 25 HOCH- ciÇ 8 Λ 30 or (b) R is hydrogen and R represents hydrogen,

CH, CH, OH OH

H ^ I I

CH3CH2-, J ^ CH- ”or CH3CH- i 37 ο

Particularly preferred are the compounds in which R

Λ is hydrogen and R is

OH

l CH ^ CH-, preferably the 5 compounds having the absolute configuration 5R, 6S, 8R.

Another preferred embodiment of the present invention comprises the preparation of the compounds in which -30

‘10 O

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

15 is -CHCH0-, -CH0CH0-, \ / or -CH0CH- | 2 2 2 yj 2 |

CH, CH

D D

Ί ο r and in which either (a) R and R taken together represent 20 <H0CHo <

O A

25 or (b) R is hydrogen and R represents hydrogen,

CH CH, OH OH

CH3CH2-, '^ CH-, ^ J> C- or CH3CH- CH ^ CH3

30, 8 Particularly preferred are compounds in which R

is hydrogen and R is * OH

CH3CH-, preferably the 35 compounds having the absolute configuration 5R, 6S, 8R.

38

A more preferred embodiment of the present invention comprises the preparation of the compounds of the formula

’T

<"> I ^ N- 0 ^ COOR ^ 10 in which -α ~ 0 15 represents cjj (I) -CH2CH2% ^) (2) -CH2CH2% ^^). CH3. ch2oh 20 © f / Vv,, ®0 Yv (3) -CH2CH2 \ _J (4) -CH2CH2 \ / ^ H3 (5) -CH2Ch / (3-CH3 (6) -CH2CH2® N ^ jV-ÆCHj 25 _ .SCH ^ (7) -ch2ch2 Ν (> η2οη (8) -CH2CH2 @ / ~ ^ ^ X) Ch3 H3Cx 30 (9) -CH2CH2 @ I ^ J) (1 °) -CH2CH2 ^ * Θ / Γ ~ λ CH3 J5CH,

(II) -CHCH2 (12) -CH2CH2®N = A

CH3 35 Diastereoisomers

R or S

39 Ί - <^ ΝΗ2 (13) -CH2CH2 I ^ _J / OR (14) Μ 'Θ diastereoisomers

5 R, R 0U S’S

I l to two asymmetric carbons of the cyclohexyl group 10 i 2 and R is hydrogen, an anionic charge or a conventional easily detachable decarboxyl protecting group, p provided that when R is hydrogen or a protecting group, a counter anion is also present, and pharmaceutically acceptable acid addition salts thereof.

14

Another preferred class of substituents R in quaternary form can be represented by the general formula: <0 1 f in which R is chosen from the group consisting of the following substituted or unsubstituted groups: alkyl, alkenyl and alkynyl, having from 1- 10 carbon atoms; cycloalkyl and cycloalkylalkyl, having 3-6 carbon atoms in the cycloalkyl nucleus, and 1-6 carbon atoms in the alkyl parts; phenyl; aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has 1-6 carbon atoms; Heteroaryl, heteroaralkyl, heterocyclyl and hetero-cyclylalkyl in which the hetero atom or atoms in the heterocyclic moieties mentioned above are selected from the group consisting of 1-4 oxygen, 40 nitrogen or sulfur atoms and the alkyl moieties associated with said heterocyclic moieties have 1-6 Λ 6 carbon atoms; in which the radicals R mentioned above are optionally substituted with 1-3 substituents independently chosen from: C 1 -C 6 alkyl optionally substituted with amino, fluoro, chloro, carboxyl, hydroxy or carbamoyl, fluoro, chloro or bromo ; 10 -OR3; -0C02R3; -0C0R3; 15 3 4 -OCONR ^ R; -0S02R3; 20 “oxo; -NR% 4; R3C0NR4-; 25 ’-NR3C02R4; -NR3C0NR3R4; -NR3S02R4; 3 ° -SR ^; 0-1-R9; 0 0 35 -S-Ry 41 -S03R3; -C02R3; 5 -C0NR3R4; -CN; or phenyl optionally substituted with 1-3 fluoro, 3 3 4 chloro, bromo, C ^ -Cg alkyl, -OR, -NR R, -SO ^ R3, -C0oR3 or -CONR3R ^ j in which R3, A 2 Q ^ 10 R and R in such substituents are as defined above; - Ί 6 or R may represent a divalent phenylene group or C 1 -alkylene, bonded to ring 15 so as to form a bridged polycyclic group and represents a non-aromatic heterocyclic radical (which may be fused or condensed with another aromatic ring Substituted or unsubstituted mono-, bi'- or polycyclic) containing at least one nitrogen in the nucleus and linked to A by nitrogen in the nucleus, thereby forming a quaternary ammonium group. The heterocyclic radical can be saturated or unsaturated (with 1-2 double bonds) and can contain up to 2 additional hetero atoms in addition to the quaternary nitrogen, such additional hetero atoms being chosen from 0, S (0) m , N, Nr "* 3 or NR ^ R ^ 8 in m 15 which m is equal to 0, 1 or 2, R is hydrogen, an alkyl group in 0 ,, - 0 / - optionally substituted or a * ^ r 17 Ί 8 35 optionally substituted phenyl and R and R are each 42 independently an optionally substituted C 1 -C 6 alkyl group or an optionally substituted phenyl.

In a preferred embodiment \ / © "Λ 5 -Ό represents a heterocyclic ring containing N non-aromatic having from 4 to 7 members, preferably 5 or 6 members, 10 containing 0-2 double bonds and 0-2 additional hetero atoms chosen from 0, S (o) m, N, NR ^ or in; 111 15 where m is equal to 0; 1 or 2, R is hydrogen, a C 1 -C alkyl group / - optionally substituted by D x 34 1-2 substituents independently chosen from -OR- ^, -NR R, 3 3 15 -002 ^ · ^ oxo, phenyl, fluoro, chloro, bromo, -SO ^ R and -CONR ^ R or phenyl optionally substituted by 1-3 substituents independently chosen from a C 1 -C 10 alkyl group -OR ^, -NR ^ R ^, fluoro, chloro, bromo, -S0XR ^, ^ U ΛΠ -io -CO ^ i and - CONR ^ R and R 'and R are each independently a C ^ -Cg alkyl group optionally substituted by 1-2 substituents independently chosen from -OR · ^, -NR ^ R, 3 3 -C0oR, oxo, phenyl, fluoro , chloro, bromo, -SOJR and -CONR R or phenyl optionally substituted with 1-3 subst ituants independently selected from a C 25 alkyl group. —O / -, -OR ^, -NR ^ R ^, fluoro, chloro, bromo, -SOJR ^, 2 D 1 4 ^ λ ·? -COpR and -CONR ^ R, in which and R ^ in such ^ ✓ 15 1718 heterocyclic groups NR J or NR 'R are such that 1 f \ defined above in relation to the substituent R.

In such a preferred embodiment, the -o nucleus may be optionally substituted with 1-3 substituents independently chosen from 43 (a) C1-C6 alkyl optionally substituted with 1-2 substituents independently chosen from fluoro, chloro, bromo, -OR3, -OCOR3, -0C0NR3R4, oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -nr3so2r4, 5 -SR3, -SO ^ R3, -C02R3 and -CONR3R4; (b) C2-C8 alkenyl optionally substituted with 1-2 substituents independently chosen from 3 3 3 a fluoro, chloro, bromo, -OR, -OCOR ^, -OCONR RH, oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -nr3so2r4, -SR3, -SO ^ R3, -COR3 and -CONR% 4; 5 (c) C ^ Cg alkynyl optionally substituted with 1-2 substituents independently chosen from 3 3 3 4 fluoro, chloro, bromo, -0RJ, -OCOR, -OCONR ^ R ^, oxo, -NR3R4, -NR3COR4, - NR3CONR3R4, -nr3so2r4, -SR3, -SO ^ R3, -C02R3 and -CONR3R4; (d) C ^ -Cg cycloalkyl optionally substituted with 1-2 substituents independently chosen from fluoro, chloro, bromo, -OR3, -OCOR3, -0C0NR3R4, oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -nr3so2r4, 20 - SR3, -SOÿt3, -C02R3 and -CONR3R4; (e) cycloalkylalkyl having 3-6 carbon atoms in the cycloalkyl nucleus and 1-6 carbon atoms in the alkyl part, optionally substituted with 1-2 substituents independently chosen from fluoro, chloro, bromo. -OR3, -OCOR3, -0C0NR3R4, oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -nr3so2r4, -SR3, -SOÿl2, -C02R3 and CONR3R4; (f) heteroaryl in which the hetero atom or atoms are chosen from the group consisting of 30 1-4 oxygen, nitrogen or sulfur atoms, optionally substituted by 1-2 substituents independently chosen from fluoro, chloro, bromo, -OR3, -OCOR3, -0C0NR3R4, oxo, -NR3R4, -NR3COR4, -NR3CONR3R4, -NR3S0oR4, -SR3, -SO, R3, -CO ~ R3 7 ^ O fs * “* 35 and -CONR ^ R ^; preferred heteroaryl radicals 44 are 5 or 6 membered aromatic heterocyclic rings; (g) heteroaralkyl in which the hetero atom or atoms are chosen from the group consisting of 5 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part has from 1-6 carbon atoms, optionally substituted by 1-2 substituents independently chosen from fluoro, chloro, bromo, -OR3, -OCOR3, -0C0NR3R4, oxo, -NR3R4, -NR3C0R4, 10 -NR3C0NR3R4, -NR3S09R4, -SR3, -S0, R3, -C05R3 i and -C0NR R; the preferred heteroaralkyls are those in which the heteroaryl radical is an aromatic heterocyclic ring with 5 or 6 members and the alkyl part has 1-2 carbon atoms; (H) 'heterocyclyl in which the hetero atom (s) are chosen from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms, optionally substituted by 1-2 substituents independently chosen from fluoro, chloro, bromo , 20 -OR3, -OCOR3, -0C0NR3R4, oxo, -NR3R4, -NR3COR4, -NR3C0NR3R4, -NR3S09R4, -SR3, -S0, r3, -C0oR3 7 A ^ ^ and -C0NR "R; heterocyclyl are preferred 5 or 6 membered saturated or unsaturated nuclei; (i) heterocyclylalkyl in which the hetero atom (s) are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl part has 1- 6 carbon atoms, optionally substituted by 1-2 substituents independently chosen from fluoro, chloro, bromo, -OR3, -OCOR3, 30 -0C0NR3R4, oxo, -NR3R4, -NR3COR4, -NR3C0NR3R4, -NR3S02R4, SR3, -S03R3, -C02R3 and -C0NR3R4; the preferred heterocyclylalkyls are those in which the heterocyclyl part is a 5 or 6 membered saturated or unsaturated nucleus; * 45 (ô ) fluoro, chloro or bromo; (k) -OR3; (1) -0C02R3; (m) -OCOR3; 5 (n) -OCONR3R4; (o) -0S02R3; (p) oxo; (q) -NR3R4; (r) R3CONR4-; 10 (s) -NR3C02R4; = (t) -NR3C0NR3R4; (u) -NR3S02R4; (v) -SR3 (w) 0 g 15 -S-R ^;

(χ) = P

-s-Ry; (y) “SO R3; (z) -C02r3; 20 (aa) -CONR3R4; (bb) -CN; or (cc) phenyl optionally substituted with 1-3 fluoro,

x L

chloro, bromo, C 1 -C 6 alkyl, -OR, -NR R, -SO, R3, -C0oR3 or -C0NR3R4. u d τ L q 25 The substituents R, R and R ^ mentioned above are as defined in relation to the

AT

substituting R.

The nucleus c.

30 __1N J

as defined above is a non-aromatic heterocyclic group. This ring, however, can be fused or fused with another ring which can be a saturated or unsaturated carbocyclic ring, preferably a C 4 -C 6 carbocyclic ring, a phenyl ring, a heterocyclic ring (saturated or unsaturated) with 4-7 members containing 1-3 hetero atoms chosen from 0, N, S (0), NR ^ or 17 1R ^ NR 'R or an aromatic ring with 5-6 members containing 5 1-3 hetero atoms chosen among 0, S (0) m, N, NR ^ or NR ^ R ^ 8 in which m, R ^, R ^ and r "'8 are as defined above.

16 14

The substituent R of the non-aromatic radical R can be either (a) an optionally substituted following group: C 1 -C 6 alkyl> C 1 -C alkenyl; G ^ -C ^ q alkynyl, C ^ -Cg cycloalkyl G ^ -Cg cycloalkyl - C ^ -Cg alkyl, phenyl, phenyl-G ^ -Cg alkyl, phenyl-C ^ -Cg alkenyl, phenyl C 2 -C 6 alkynyl heteroaryl, heteroaralkyl in which the alkyl part has from 1-6 carbon atoms, heterocyclyl or heterocyclylalkyl in which the alkyl part has from 1-6 carbon atoms is (b) a phenylene or alkylene group in C ^ -C ^ divalent linked to the nucleus

-O

so as to form a polycyclic group with a bridged nucleus, for example a quinuclidine group. The heteroaryl substituent (or the heteroaryl portion of heteroaralkyl) may be a mono-, bi- or polycyclic aromatic heterocyclic group containing 1-4 atoms of 0, N or S; 5 or 6 membered heterocyclic rings are preferred, such as thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, tetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyrididyl pyridinyl. The heterocyclyl substituent (or the heterocyclyl portion of heterocyclyl-j-alkyl) may be a mono-, bi- or polycyclic saturated or unsaturated non-aromatic heterocyclic group containing 47 1-4 atoms of 0, N or S; 5 or 6 membered heterocyclic rings are preferred, such as morpholinyl, piperazinyl, piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrrolinyl and pyrrolidinyl, 5 In the case where the substituent R ^ is an alkyl, alkenyl, alkyl group, cycloalkyl cycloalkylalkyl, phenyl, phenylalkyl, phenylalkenyl, phenylalkynyl, heteroaryl, heteroaralkyl, heterocyclyl or heterocyclylalkyl, such groups may be optionally substituted with 1-3 substituents independently = selected from: (a) C group ^ -Cg optionally substituted by, preferably 1 -3, amino, fluoro, chloro, carboxyl, hydroxy or carbamoyl groups; (B) fluoro, chloro or bromo; (c) -OR3; (d) -0C02R3; (e) -0C0R3; (f) -0C0NR3R4; (G) -0SO2R3; (h) oxo; (i) -NR3R4; (j) R3C0NR4-; (k) -NR3C0oR4; / \ 3 2 3 4 25 (1) -NR CONR R; (m) -NR3SO2R4; (n) -SR3; (o) -SOR9; (p) -SO2R9; 3 ° (q) -SO3R3; (r) -C02R3: (s) -C0NR3R4; (t) -CN; or 1 (u) phenyl optionally substituted with 1-3 substituents independently chosen from fluoro, chloro, 48 bromo, Oj-Cg alkyl, -OR ^, -NR ^ R ^ 1, -SO ^ R- ', -CCRR ^ or -CONR ^ R ^, in which, with respect to Λ Ci to the substituents R mentioned above, the 3 4 groups R- 'and R are independently chosen from hydrogen; an alkyl, alkenyl and alkynyl group having 1-10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl, having 3-6 carbon atoms in the cycloalkyl nucleus and 1-6 carbon atoms in the alkyl parts; phenyl; aralkyl, aralkenyl = 'and aralkynyl in which the aryl part is phenyl and the aliphatic part has from 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the heteroaryl and the heterocyclyl group or the part of a group is as defined above for Λ 6 R and the alkyl parts associated with said heterocyclic parts have 1-6 atoms of carbon; 3 4 or R ^ and R taken together with the nitrogen to which at least one is bonded can form a hetero-cyclic ring (as defined above for R ^) containing nitrogen with 5 or 6 members; R ^ is as 3 defined above for R ^ except that it cannot be hydrogen. A more preferred substituent R is a C 1 -C 6 alkyl group, especially methyl.

In the case where R ^ is a divalent phenylene or alkylene group, j-Cg, such a group is linked to another atom of the nucleus 30 \ / Θ \

-O

- so as to form a bridged polycyclic ring, for example a quinuclidine ring in quaternary form of the formula: 49

U

5 I®

A particularly preferred embodiment of the present invention comprises the preparation of the compounds of formula I in which

io ITn ® J

represented

15 \ ^ ν C \ ^ YY

20 γ v 1-v% / -V "% Γ ~> <^ ®V- \ \ N7® Λ,>>“ \ ’

\ _ / v_y V-? N

25 / / / CH

/ ~ r \ c ^ n \ 3 θ \ _ / 3 ® \ _ / ® \ _ / CH3 30 CH * -CQ "UÜ 50 in which Y is hydrogen, a C ^ -Cg alkyl group, hydroxy, -S-C 1 -C 6 alkyl> carboxyl, carbamoyl, chloro, bromo, iodo, fluoro or phenyl In this subclass, the preferred compounds are those in which A is -CHCH0-, - / \, -CH9CH- or i 2 \ _y 2i CHj y- CH3 n - (CH ^^ - in which n is 2, 3 or 4, more preferably 10 in which A is -CHgC ^ -, -QE ^ CHgCHg- "- CHCH0-, - / \ or -CH9CH-

I 2 2I

ch3 y ch3 15 and more preferably those in which A is -CH9CH9-, 18 ^ ^ ^ and in which either (a) R and R taken together represent H0CHo> 2C = 20 CH, 3

O

either (b) R is hydrogen and R represents hydrogen,

ch3 ch3 OH OH

CH, CH0-, ^ CH-, ^ C- or CH, CH- 5 ^ ^ 3 25 CH ’CH, 5 5 8

Particularly preferred are the compounds in which R

, „1 is hydrogenated and R is

OH

CH3CH-, especially compounds having the absolute configuration 5R, 6S, 8R.

An even more preferred embodiment of the present invention comprises the preparation of the 51 compounds of formula I wherein r1Lq 5 represents: CH, ______ CH3 / \ £> \ 10 _J \ _ / CH, y-- CH, y -, CH,, -v CH, N3 / \ X / \ N3 / \ ®X 3 15 —s— * · 0, —N N-CH, —-N ï <© \ _ / © \ _ / 3 © \ _ / \ ch3% / - \ cÎbZ \ ® /> ^ = \ 20 \ n © â, —1f f1 or /; v_v w

In this preferred subclass, preferred compounds are those where A is -CHCH2-? - / \, CH2CH or CH, / 'CH, 5 3 ”^ CH2 ^ n“ in which A is 2, 3 or 4, more preferably those in which A is -CH ^ CHg-, -CH ^ CHgCHg-, - (j! HCH2-, -ί \ or -CH2CH-, ch3, ch3 35 and more preferably those in which A is -CH9CH9-, and in which either (a) R and R taken together represent: 52 HOCHp <C = a λ 5 either (b) R is hydrogen and R represents hydrogen,

ch3 ch3 OH OH

CH3CH2-, ^ CH- or CH3CH-

CrT CH, - 3 3 ^ ^ g 10 The compounds in which R 5 is hydrogen and R is particularly preferred

OH

CH ^ H-, especially the compounds having the absolute configuration 5R, 6S, 8R.

An even more preferred embodiment of the present invention comprises the preparation of the compounds of formula I in which: 20 V __ / represents:

25 Y

wherein Y is hydrogen, C 1-6 alkyl, hydroxy, -S-C 1-6 alkyl, carboxyl, carbamoyl, chloro, bromo, iodo, fluoro or phenyl. In this preferred subclass, the preferred compounds are those in which A is - (CH2) n ~ in which n is 2, 3 or 4, more preferred are those in which A is 35 -CH2CH2- and in which either (a) R and R taken together 53 represent:

HOCH

> CH3 5 8, either (b) R is hydrogen and R represents hydrogen,

ch3 CH OH OH

CH3CH2-, ^ .CE-, or CH3CH

CH3 CH3 * 10

Particularly preferred compounds are those in, g Λ * where R is hydrogen and R is

OH

CH3CH-, especially compounds having the absolute configuration 5R, 6S, 8Ro

A more preferred embodiment of the present invention comprises the preparation of the compounds of formula I in which:

20 R x-nO

represents 25 CH, H)

In this preferred subclass, preferred compounds are those in which A is - (CH2) - in which n is 2, 3 or 4, more preferred are those in which A is -CH2CH2- and in which either ( a) r "^ and R® taken together represent HOCHo ch3 54 8 Λ

either (b) R is hydrogen and R represents hydrogen, CH * CH * OH OH

CH3CH2-, J ^ CH-, or CH3CH

ch3 ch3 5

Particularly preferred are the compounds in which R® is hydrogen and r "* is

OH

CH3CH-, 10 I and especially the compounds having the absolute configuration 5R, 6S, 8R.

A more preferred embodiment of the present invention comprises the preparation of the compounds of the formula

OH H

A 'CK

(R) "ΓΠ | pS-CH2CH2-N®J

20 0 ^ N- COOR ^ in which »A Ç \ CH- *

25 R1xb / 'N X

—N) represents —N

30 C ^ 3 / \ ^ r ~ \ \ 3ΛΛ -NS, - NP f- 0 © \ _ / ®W ^ '(both diastereoisomers 35 c * and / ¼) 55 ch3, -v eu * / - \ ch3

- N N-CH_ or —N N

Θ \ _ / 3 © \ _ / ^ CH3 5 p and R is hydrogen, an anionic charge or a readily separable conventional decarhoxyOe protecting group, 2 provided that when R is hydrogen or a protecting group, a counter ion or contrary ion is also present, and pharmaceutically acceptable acid addition salts thereof.

It will be appreciated that certain products within the scope of formula I can be formed as optical isomers as well as as epimeric mixtures thereof. It is understood that the present invention encompasses all of these optical isomers and these epimeric mixtures. For example, when the substituent at position 6 is hydroxyethyl, such a substituent can be either in the R configuration or in the S configuration and the resulting isomers as well as the epimeric mixtures thereof are included in or encompassed by the present invention .

The process of the present invention uses the formula: 25; i8 h 4τγ 30 \ C00R2 ’

IV

56 which has been described, for example, in European patent application 38,869 and which can be prepared by the general methods described therein. L represents a conventional separable group (defined as "X" in European patent application 38 869) such as chloro, bromo, iodo, benzenesulfonyloxy, p-toluenesulfonyloxy, p-nitro-benzenesulfonyloxy, methanesulfonyloxy, trifluoromethane-sulfonyloxy, diphenoxyphosphinyloxy or di (trichloroethoxy) - ^ phosphinyloxy. The preferred separable group is the group

10 diphenoxyphosphinyloxy. Intermediaries of formula IV

j, are generally formed in situ by reacting the intermediate ♦ of the formula: r8 H o 15 R1_ \ C00R2 '

III

20 18 2 'in which R, R and R are as defined above, with an appropriate acylating agent R ° - L. The preferred intermediate IV where L is a diphenoxyphosphinyloxy group can be prepared by reacting the keto ester III in an inert organic solvent such as methylene chloride, acetonitrile or dimethylformamide with approximately an equimolar amount of diphenyl chloro-phosphate in the presence of a base such as diisopropylethylamine, triethylamine, 4-dimethylamino -30 pyridine or the like, at a temperature of about -20 ° C to + 40 ° C, more preferably about 0 ° C. Intermediate IV can be isolated, if desired, but is suitably used as a starting material for the process of the present invention without isolation or purification.

57

In the present process, the carbapenem IV intermediate is reacted with a quaternary thiolamine compound of the formula: 14 HS — A — R1 ^ r 5 #

VII

in which A is a cyclopentylene, cyclohexylene or C 1 -C 6 alkyl group optionally substituted by one or more C 1 -C 4 alkyl groups, more preferably 10 cyclopentylene, cyclohexylene or R10 R12 ê l_! ~ A11 to "15 in which R ^, R ^, R ^ and R ^ are each independently hydrogen or a C ^ -C ^ alkyl group, is a counter anion associated with a strong acid, such as Cl", Br “, CH ^ SO ^“, CF ^ SO ^ - or 20 CH3- ^ r ”^ - S03- and 14 R is an aromatic or non-aromatic heterocycle containing nitrogen in quaternary form as defined above. The reaction is carried out in an inert solvent such as acetonitrile, acetonitrile-dimethylformamide, tetrahydrofuran, tetrahydrofuran-H 2 O, acetonitrile-2 O or acetone in the presence of a base. The nature of the base is not critical. Best results, however, have been obtained when a non-nucleophilic tertiary amine base such as diisopropylethylamine, 1,8-

diazabicyclo [5.4.0l undec-7-ene, 1,5-diazabicyclo [4.3.0J

-non-5-ene or a trialkylamine such as triethylamine, tributyiamine or tripropylamine is used. The reaction of intermediate IV thiol VII can be carried out with a wide temperature range, for example 58 of -15 ° C to room temperature, but preferably carried out at a temperature in the range from about -15 ° C to + 15 ° C, more preferably around 0 ° C.

The carbapenem product produced by the reaction of the thiol VII of the quaternary amine with the intermediate IV will have a counter anion associated with it £ for example (CgH ^ 0) 2PO2®, Cl® or the anion associated with the quaternary thiol ^ which can at this stage be replaced by a counter anion -? different, for example one which is more pharmaceutically acceptable, by conventional procedures. Alternatively, the counter anion can be removed during the subsequent unblocking step. If the carbapenem compound in quaternary form and the counter anion form an insoluble product, the product can crystallize as it is formed and be collected pure by filtration.

Following the formation of the desired 2 'carbapenem product, the carboxyl protecting group R of compound 1' can optionally be removed by conventional procedures such as solvolysis, reduction or chemical hydrogenation. If a protecting group such as p-nitrobenzyl, benzyl, benzhydryl or 2-naphthylmethyl is used which can be removed by catalytic hydrogenation, the intermediate 1 'in a suitable solvent such as dioxane-water-etiole, tetrahydrofuran-diethyl- buffer ether, tetrahydrofuran-aqueous dipotassium hydrogen phosphate-isopropanol or the like can be treated under hydrogen pressure from 1 to 4 atmospheres in the presence of a hydrogenation catalyst such as palladium on charcoal, palladium hydroxide, platinum or the like at 0 to 50 ° C for about 0.24 to 4 hours. When R is a group such as o-nitrobenzyl, photolysis can also be used for deblocking. Protecting groups such as 2,2,2-trichloroethyl can be removed by reduction with mild zinc. The allyl protecting group can be removed using a catalyst comprising a mixture of a palladium compound and triphenylphosphine in a suitable aprotic solvent such as tetrahydrofuran, methylene chloride or diethyl ether. Similarly, other conventional carboxyl protecting groups can be removed by methods known to those skilled in the art. Finally, as mentioned above 2 '

above, compounds of formula 1 'where R is a physiologically hydrolyzable ester such as acetoxymethyl, phthalidyl, indanyl, pivaloyloxymethyl, methoxymethyl, etc. can be administered directly to the host without release since such esters are hydrolyzed in vivo under physiological conditions. It will be understood that if the substituent R and / or R

or the substituent R1 ^ nucleophile in quaternary form linked to the substituent A contains a functional group which could interfere with the course of the expected reaction, such a group can be protected by a conventional blocking group and then be subsequently unblocked to regenerate the group desired functional. Appropriate blocking groups and suitable procedures for the introduction and removal of such groups are well known to those skilled in the art.

As in the case of other antibiotics / 1-lactam, compounds of general formula I can be converted by known procedures into pharmaceutically acceptable salts which, for the purposes of the present invention, are substantially equivalent to compounds not salty.

Thus, for example, one can dissolve a compound of formula I wherein R is an anionic charge in a suitable inert solvent and then add an equivalent of a pharmaceutically acceptable acid. The desired acid addition salt can be recovered by conventional procedures, for example by solvent precipitation, lyophilization, etc. If other basic or acidic functional groups are present in the compound of formula I, pharmaceutically acceptable basic addition salts and pharmaceutically acceptable acid addition salts can be similarly prepared by methods known.

2

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

Some of the thiol intermediates of formula VII can be prepared, for example, by reacting a sulfide of the formula: ό d - 25 Villa VlIIb R1 2 —C —— C — R3 30

City Λ Λ 12 Λ ^ 2 in which R, R, R and Ru are each independently hydrogen or a C ^ -C ^ 3 alkyl group with a heteroaromatic amine (as defined above) 35 of formula 61 Ο or a non-aromatic heterocyclic amine (as defined above) of the formula <o 3 10 and a strong acid. The reaction can be carried out in the presence or in the absence of an inert organic solvent which is preferably a non-polar organic solvent such as methylene chloride, benzene, xylene, toluene or the like. If the amine and the sulfide reagents are liquids or if a solid amine is soluble in a liquid sulfide reagent, it is preferred to carry out the reaction without the use of an additional solvent.

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

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

The sulfide reagent, the aromatic amine and the acid are preferably used so that the sulfide and acid are used in approximately equimolar amounts with the amine being used in excess, for example two or three moles of amine per mole of sulfide or acid.

a The quaternary thiol amine intermediate will have a counter anion associated with it which will be determined by the particular acid employed. Of course, it is possible at this time to substitute a different counter anion by conventional procedures for use in the subsequent reaction with the intermediate carbapenem IV.

5 The carbapenem derivatives of the general formula I

wherein R is hydrogen, an anionic charge or a physiologically hydrolyzable carboxyl protecting group, or the pharmaceutically acceptable salts thereof, are potent antibiotics active against various gram-positive and gram-negative bacteria and they * can be used, for example, as animal feed additives to promote growth, as preservatives in food, as bactericides in industrial applications, for example in water-based paint and in water bleaching ("white water") paper rolls to inhibit the growth of harmful bacteria and as disinfectants for the destruction or inhibition of the growth of harmful bacteria on medical equipment and den-20. They are especially useful, however, in the treatment of infectious diseases in humans and other animals caused by gram-positive or gram-negative bacteria.

The pharmaceutically active compounds provided by the novel process of the present invention can be used alone or formulated as pharmaceutical compositions comprising, in addition to the active carbapenem ingredient, a pharmaceutically acceptable carrier, diluent or excipient. The compounds can be administered by a variety of routes or means; those of main interest include: an oral, topical or parenteral route (intravenous or intramuscular injection). The pharmaceutical compositions can be in solid form such as capsules, tablets, powders, etc., or in liquid form such as solutions, suspensions or emulsions. Compositions for injection, the preferred route of administration, may be prepared as a unit dose in ampoules or in multi-dose containers and may contain formulating agents such as suspending, stabilizing and dispersing agents. The compositions can be in ready-to-use form or in powder form for reconstitution at the time of administration with a suitable vehicle such as sterile water.

The dosage to be administered largely depends on the particular compound to be used, the particular composition formulated, the route of administration, the nature and condition of the host and the particular site and organism to be treated. The selection of the particular preferred dosage and route of application is then left to the discretion of the therapist or practitioner. In general, however, the compounds can be administered parenterally or orally to mammalian hosts in an amount ranging from about 5 to 200 mg / kg / day. Adminis-20 tration is usually done in divided form, for example three to four times a day.

Other objects, characteristics and advantages of the present invention will become apparent in the light of the explanatory description which follows, given with reference to the following examples, given simply by way of illustration and which therefore cannot in any way limit the scope of the invention.

64 EXAMPLE 1

Preparation of 3-f2- (1-pyridinium) ethylthioj- 6 <X -Cl- (R) -hydroxyethvlT-7-oxo-1-azabicyclo (5.2.0) - hept-2-ene-2-carboxylate 5 A. 1- (2-Mercaptoethyl) pyridinium methanesulfonate "A + / ô \ + Ms0H _HS Ks0 10 \ ~ y 55 ° C, 16 h At a suspension of pyridinium methanesulfonate in pyridine prepared by dropwise addition 15 d methanesulfonic acid (1.95 ml, 0.03 mol) to pyridine (8.0 ml, 0.099 mol) with cooling, ethylene sulfide (1.96 ml, 0.033 mol) is added. The resulting mixture is stirred at 55 ° C. for 16 hours and concentrated under reduced pressure into a thick syrup which is mixed with a few milliliters of water The solution is poured onto the top of a column (40 x 15 cm) of micro-bondapak C- 18 (trade name) which is eluted with water. Lyophilization of the appropriate fractions gives a colorless syrup, 6.5 g (9V / o), IR (film) ύ:

IïlaX

25 2300-2600 (br, SH), 1635 (pyridinium), 1490, 1200 (suifonate), 1068, 1060, 1045, 791, 780 cm “1, 1 H NMR (DMSO-dg) S: 2.32 (3H, s , CH3S03 "), 2.61, 2.70, 2.73, 2.82 (1H, B part of the A2B, SH system), 3.07 (2H, m [with D2O, 3.08 (2H, t , J = 6.5 Hz)}, CH2S), 4.76 (2H, t, J = 6.5 Hz, 0Η £ Ν +), 30 8.19 (2H, m, Hm of pyridinium), 8, 6 (1H, m, Ho of pyridinium), 9.08 (2H, dd, J = 6.8 Hz, J = 1.4 Hz, Ho of pyridinium), UY (H2O)> max: 206 (£ 5,230) , 258 (£ 3760) m jl,.

65

METHOD A

B. 1- (2-Mercaptoethyl) pyridinium chloride ^ / z-n \ Permutit S-1 Cl “V ~ \

Mso -► l (©) 10 Pour an aqueous solution of methane sulfonate s 1- (2-mercaptoethyl) pyridinium (9.4 g, 0.04 mole) at the top of a column (2.5 x 41 cm ) of permutit S-1 Cl ”(trade name). The column is eluted with water at a rate of 0.5 ml per minute and the appropriate fractions are combined and lyophilized to give a yellowish syrup 7.0 g (100%) which is used as it is for the step next, 1H NMR (D20) S: 3.22 (2H, m, CH2S), 4.88 (m, CH2N +), 8.18 (2H, m, pyridinium Hm), 8.7 (1H, m, Hp of pyridinium), 9.0 ppm (2H, m, Ho of pyridinium).

20 METHOD B

. Δ. <0> · (Cÿi - 25 Cl "To a precooled pyridine (ice bath (5.6 ml, 70 mmol), hydrochloric acid pyridine 30 (4.05 g, 35 mmol) and ethylene sulfide are added (2.1 ml, 35 mmol) The mixture is heated to 65 ° C and stirred for 75 minutes to give a two phase system.

The lightest phase is removed. The remaining oil is washed with ether (5 x 10 ml) and pumped under high vacuum to give the title compound (90-100%) which is used as it is for the next step.

C. Paranitrobenzyl chloride 5- C 2- (1-pyridinium) - ethylthio] -6c <- L 1 ~ (E) -hydroxyethyll -7 ~ oxo-1 ~ azabicyclo (3 ”2.0.) Hept — 2-ene— 2-carboxylate 5

OH 1 "NEt (iPr) 2 OH

2- ClP (OPh)? / ZT \ 0 ^ ° »rMÉ> S COOPNB -3 \ _ / COOPNB L1 4- NEt (iPr) 2 15

A solution of p-nitrobenzyl 6 <K- £ l- (R) -hydroxyethyl] -3,7-dioxo-1-azabicyclo (3.2.0) -heptane-2-carboxylate (6.09 g, 17, is cooled. 5 mmol) in acetonitrile (20 ml) at + 5 ° C under a nitrogen atmosphere and treated successively with diisopropylethylamine (3.65 ml, 21.0 mmol) and diphenyl chlorophosphate (4.34 ml , 21.0 mmol). The resulting mixture is stirred for 30 min at 5 ° C, cooled to -5 ° C and treated successively with a solution of crude 1- (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 hour, cooled to -30 ° C and stirred for an additional 15 minutes. The solid is filtered and washed with cold acetonitrile (-30 ° C); 5.77 g (6596), IR (nujol) v) mQx: 3300 (OH), 1775 (C = 0 of yi-lactam), 1690 (C = 0 of the PNB ester), 1630 (pyridi-nium) , 1605 (phenyl of the PNB ester), 1515 (N02), 1335 cm - "* (N02), 1H NMR (DMS0- d6) S: 1.17 (3H, d, J = 6.1 Hz, Cï ^ CHOH), 67 3.2-3.75 (5H, H-4, H-6, CH2S), 3.75-4.5 (2H, H-5, CH3CHOH), 4.92 (2H, brt , J = 6.5 Hz) CH2N +), 5.18 (1H, d, J = 4.9 Hz, OH), 5.37 (center of ABq, J & b = 14.2 Hz, CH2 of GNP), 7.69 (2H, d, J = 8.7 Hz, Ho of PNB) '8.24 (d, J = 8.7 Hz, Hm of PNB), 5 8.0-8.4 (4H, Hm of PNB, Hm of pyridinium), 8.66 (1H, Hp of pyridinium), 9.17 (2H, brd, J = 5.5 Hz, Ho of pyridinium).

The filtrate and the washing are combined and diluted with ether (150 ml). The supernatant solution is decanted and the gum is dissolved in water (40 ml) containing 10 enough acetonitrile to have a solution which is poured onto the top of a column (3 x 10 cm) of micro-bondapak C-18 (tradename). The column is eluted with mixtures of 10% acetonitrile-90% water (150 ml) and 50% acetonitrile-50% water (100 ml). The appropriate fractions are combined and lyophilized after the acetonitrile has been removed in vacuo to give a yellowish powder. An NMR spectrum of this shows the presence of the title compound mixed with a little p-nitrobenzyl diphenylsulfonate 3- ^ 2- (1-pyridinium) ethylthio 3-6 o (- [1- (R) -20 hydroxyethyl3 -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate (2: 1). The powder is dissolved in water (minimum quantity) and passed through a column (1 , 5% 21 cm) of permutit S-1C1 "(trade name) with water. Lyophilization of the appropriate fractions gives 1.8 g (20%) of the title compound.

D. Paranitrobenzyl diphenylphosphate 5- C 2- (1- pyridinium) ethylthio3 -6QÇ-Γ 1- (R) -hydroxyethvll - 7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

0H 1) NEt (iPr) 9 PH

30 T 0 2 A

Arry0 „n> sW © \ 3) Hs ~ © Ms <r Λ ^ C '° 0PHB 4) NEt (iPr) 2 (PhO) jcr 68

A solution of p-nitrobenzyl 6o (- £ 1- (R) -hydroxy-ethyl 1 -3,7-dioxo-1-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 and treated successively with diisopropylethylamine (0.105 ml, 0.60 mmol) and diphenyl choropbosphate (0.124 ml, 0.60 mmol) .

The resulting solution is stirred for 30 minutes at 0 ° C and treated successively with a 1- (2-mercaptoethyl) pyridinium methane sulfonate solution (0.170 g, 0.72 mmol) in acetonitrile (0.6 ml) and diisopropyl-. ethylamine (0.105 ml, 0.60 mmol). The reaction mixture is stirred at 0 ° C for 15 minutes, diluted with cold water (0 ° C) (7 ml) and poured onto the top of a column (1.5 x 6.4 cm) of micro-bondapak C-18 (trade name). The column is eluted with a mixture of acetonitrile (25% -50 <X)) in water (75% -50%). The appropriate fractions are combined and lyophilized after the acetonitrile has been removed in vacuo to give a yellowish powder 0.33 g ($ 92; IR (KBr) Omax- 3600-3000 (OH), 1765 (C = 0 of / -lactam), 1690 (C = 0 of the PNB ester), 1625 (pyridinium), 1585 (phenyl), 1510 (NO2), 1330 (NO0), 885 cm-1 (NO2), NMR 1H (DMSO-dg) ^: 1.16 (3H, d, J = 6.2 Hz, CH ^ CHOH), 4.87 (2H, brt, J = 6.6 Hz, CH2S), 5.37 ( center of ÀBq, Ja ^ = 14.3 Hz, CH2 of PNB), 6.7-7.5 (phenyl), 25 7.68 (d, J = 8.8 Hz, Ho of PNB), 8.23 (d, J = 8.8 Hz, Hm of PNB), 8.0-8.3 (m, Hm of pyridinium), 8.4-8.8 (1H, Hp of pyridinium), 9.09 (2H , dd, J = 6.7 Hz, J = 1.3 Hz, Ho of pyridinium).

E. 3- C2- (1 -pyridinium) ethylthioH -6oc - Cl- (R) -hvdroxv- ethyl3 -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxvlate

Method A

69 0H] H / -v cr b, ff 0 coo “5 COOPNB (ph0) 2 | o- To a solution of diphenylphosphate 10 p-nitrobenzyl 3-L 2- (1-pyridinium) ethylthio 1 -6 of- 11 - (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3. 2.0) hept-2-ene-2-carboxylate (0.16 g, 0.22 mmol) in wet tetrahydrofuran (10 ml) is added ether (10 ml), monobasic potassium phosphate-sodium hydroxide buffer pH 7.4 (16 ml, 0.05 M) and 10% palladium on carbon (0.16 g). The resulting mixture is hydrogenated under a pressure of 275.792 kPa for 1 hour at 25 ° C. The two phases are separated and the organic phase is extracted with water (2 times 3 ml). The aqueous solutions are combined, washed with ether (2 times 10 ml) and poured at the top of a column (1.5 x 6.2 cm) of micro-bondapak C-18 (trade name ) after the traces of organic solvents have been removed under vacuum. Elution of the column with water gives, after lyophilization of the appropriate fractions, a yellowish powder 0.062 g (84%), IR (KBr) ^ max: 3700-3000 (OH), 1755 (C = 0 of y ^ -lactam), 1630 (pyridinium), 1590 cm “1 (carboxylate), 1H NMR (D20): 1.22 (3H, d, J = 6.4 Hz, CH3CH0H), 2.92 (d, J = 9 , 1 Hz, H-4), 2.97 (d, £ = 9.1 Hz, H-4), 3.20 (dd, J = 2.5 Hz, J = 6.1 Hz, H-6 ), 30 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, C ^ N4-), 8.04 (m, Hm of pyridinium), ·? ! 70 8.5 (m, pyridinium Hp), 8.82 (dd, J = 3.2 Hz, J = 1.1 Hz,

Ho of pyridinium), UV (H_0) X: 259 (£ 5800), max 296 (6 7030) m dd, T ^ = 13.5 h (measured at a concentration of 10 M in phosphate buffer pH 7.4 at 36.8 ° C).

5

Method B

fH 10/0 Pd / T / 7T \ "j" -. o ^ 'T. w

C00PNB

15 To a solution of p-nitrobenzyl chloride 3- | _2- (1-pyridinium) ethylthio3 -6 "- Q1- (R) -hydroxyethyl3 -7-oxo 1-azahicyclo (3.2.0) hept-2-ene -2-carhoxylate (5.77 g, 11.4 mmol) in monobasic potassium phosphate-sodium hydroxide buffer (170 ml, 0.2 M, pH 7.22), tetrahydrofuran (30 ml) is added, ether (30 ml) and Λ0% palladium on charcoal (5.7 g). The resulting mixture is hydrogenated at 22 ° C under a pressure of 275.792 KPa 25 for 1 hour and filtered through a pad of Celite (trade name). The tampon is washed with water (2 x 15 ml). The filtrate and the washes are combined and diluted with ether (100 ml). The aqueous phase is separated, washed with ether (3 times 100 ml) and poured onto the top of a column (4.5 x 20 cm) of micro-bondapak C-18 (trade name) after the organic solvents have been removed under vacuum. Elution from the column with water followed by a mixture of 1% acetonitrile in water gives lyophilization of the appropriate fractions 35 2.48 g (65%) of the title compound as a powder 71 yellowish. The analytical data was identical to that reported for the compound prepared according to method A.

EXAMPLE 2 5 Preparation of 5- L 2- (1- (3,5-dimethylpyridinium) - ethylthio 1 -6Q (- Cl - (R) -hydroxy thyl1 -7-oxo-1-azabicyclo (3 "2,0) hept-2-ene-2-carboxylate DE CH * 10 f _ / 3 èoo- 15 A. 1- (2-Mercaptoethyl) -3.5- dimethylpyridinium methanesulfonate 2 ° g - / j- / / \ + N Çj \ + MsOH - * ©) MS ° 25 To a suspension of 3,5-lutidinium methanesulfonate in 3,5-lutidine prepared by the addition of methanesulfonic acid (0.65 ml, 0.010 mol) to 3, Cold 5-lutidine (2.51 ml, 0.022 mole) ethylene sulfide (0.655 ml, 0.011 mole) is added, the resulting mixture is stirred under nitrogen at 55 ° C for 24 hours, cooled to 23 ° C. and diluted with water (5 ml) and ether (5 ml). The organic layer is separated and the aqueous solution is washed with ether (6 times 4 ml). trace the ether in vacuo and apply the solution to the top of a column (2.5 x 6.0 cm) 72 of micro-bondapak C-18 (trade name). Elute the column with water and lyophilization of the appropriate fractions gives 2.4 g of colorless syrup (91%); I.R. (film) \): 2520 (SH), 1628 (pyridinium), 1600, 1495, 5 1325, 1305, 1283, 1200 (sulfonate), 1040, 938, 765, 680 cm'1; 1H NMR (DMSO d6) <5 ": 2.31 (3H, s, CHjSO ^), 2.47 (6H, s, CH ^ on pyridinium), 2.57, 2.66, 2.69, 2, 78 (1H, B part of the system A ^ B, SH), 3.06 (2H, m [with D ^ O added (2H, t, J = 6.5 Hz)], CH2S), 4.65 (2H , t, J = 6.5 Hz, CH2N +), 8.34 10 (1H, s, Hp of pyridinium), 8.79 (2H, s, Ho of pyridinium);; UV (H20) A.max: 271 (£ 4860) mju .. Anal hold, for C10H17N03S2.0.5H20: 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 B. Paranitrobenzyl diphenyl phosphate 3-12- (1- (5,5-dimethylpyridinium)) ethvlthiol -6oç - Cl- (R) -hydroxyethyl 1 -7-oxo-1-azabicyclo (3.2.0) hept-2- ene-2-carboxylate 20

1) NEt (iPr) Q

OH 2 OH

AT 2) ClP (OPh) / tA

4) NEt (iPr) 2 30 A cold solution (0 ° C) of p-nitrobenzyl 6 4f- (1- (R) -hydroxyethyl) -3,7-dioxo-1-azabicyclo (3.2.0) heptane- 2-carboxylate (0.523 g, 1.50 mmol) in acetonitrile (6.0 ml) maintained under a nitrogen atmosphere is added; diisopropylethylamine (0.314 ml, 1.8 mmol) followed by diphenyl chlorophosphate (0.373 ^ 1.1.8 mmol).

1 73

The reaction mixture is stirred for 30 minutes and treated with a solution of 1- (2-mercaptoethyl) -3,5-dimethylpyridinium methanesulfonate (0.493 g, 1.87 mmol) in acetonitrile (1.9 ml) followed with 5-diisopropylethylamine (0.314 ml, 1.8 mmol). The reaction mixture is stirred at 0 ° C for 1 hour, diluted with cold water (° C) (26 ml) and poured onto the top of a column (7.0 x 3.5 cm) of micro- bondapak C-18 (trade name). Elution from the column with a mixture of 25-50% acetonitrile-75-50% water gives after lyophilization of the appropriate fractions 1.01 g (90%) of the title compound;> as a yellowish powder, IR (KBr) ^ max: 3700-3100 (OH), 1778 (C = 0 of yi-lactam), 1700 (C = 0 of the PNB ester), 1635 (pyridinium), 1595 (phenyl) , 1521 (N02), 15 1335 (N02), 895 cm ”1 (N02), 1H NMR (DMSO dg) S: 1.16 (3H, d, J = 6.1 Hz, CH ^ CHOH), 2, 43 (s, CH ^ on pyridinium), 4.75 (2H, m, CH2N +), 5.38 (center of ABq, fe = 14.3 Hz, CH2 of PNB), 6.6-7.5 (10H , m, phenyl), 7.70 (2H, d, J = 8.7 Hz,

Ho of PNB), 8.0-8.5 (3H, m, Hp of pyridinium, Hm of PNB), 20 8.82 (2H, s, Ho of pyridinium), UV (H20) Xmaxï 270 (£ 11,570) , 306 (£ 7343) πyjo. Anal, hold, for C37H38N3 ° 10SP‘H20: 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.

C. 3- C2- (1- (3,5-dimethylpyridinium)) ethylthio3 -6 ^ - 25 Cl- (R) -hydroxyethyl2-7-oxo-1-azahicyclo (3.2.0) hept-2 -ene-2 -carboxylate | H / 10% Pd / C, F, - (30 y **, - / pj \ H2 ο Θ> THF, ether, '* 0' '' 'v \ Π buffer \ COOPNB (Ph0) 2P0 " C00 ”4 74 To a solution of p-nitrobenzyl diphenylphosphate 3- [2- (1- (3,5-dimethylpyridinium)) ethylthio ~] -6 <X- Y1- (R) -hydroxyethylJ -7-oxo-1 -azabicyclo (3.2.0) hept-2-ene-2-carboxylate (0.600 g, 0.80 mmol) in wet tetra-5 hydrofuran (36 ml) added ether (36 ml), buffer monobasic potassium phosphate-sodium hydroxide (0.05 M, pH 7d4, 44 ml) and 10% palladium on carbon (0.60 g) The resulting mixture is hydrogenated at> a pressure of 275.792 kPa at 23 ° C for 1.25 hours.

The organic layer is separated and extracted with buffer i (2 times 5 ml). The aqueous layers are combined, filtered through a pad of Celite (trade name), washed with ether (40 ml), pumped to remove traces of organic solvents and poured on top of a 15 column (2.5 x 10.0 cm) of micro-bondapak C-18 (trade name). Eluting the column with water and lyophilizing the appropriate fractions gives the title compound 0.186 g (64%) as a yellowish powder IR (KBr): 3700-3100 (OH), 1760 (c = 0 of Â-lactam), IîlcLX. * / 20 1595 cm (carboxylate), NMR H (D ^ O): 1.21 (3H, d, J = 6.3 Hz, CH ^ CHOH), 2.45 (6h, s, CH ^ on pyridinium) , 2.81 (d, J = 9.2 Hz, H-4), 2.96 (d, J = 9.2 Hz, H-4), 3.22 (dd, J = 2.6 Hz, J = 6.2 Hz, H-6), 3.40 (t, J = 6.2 Hz, CH2S), 3.84 (dd, J = 9.2 Hz, J = 2.6 Hz, H- 5), 4.15 (m, CH ^ CHOH), 4.71 (t, J = 6.2 Hz, CH2N +), 8.21 (1H, s, pyridinium Hp), 8.46 (2H, s, pyridinium Ho), UV (H2 °) Xmax: 279 (£ 8,345), 296 (£ 7,714) m, MD23 + 40.7 (c 0.53, Η £ 0) = 16.9 h (pitted at a concentration of 10 ~ ^ M phosphate buffer 30 pH 7.4 at 36.8 ° C).

75 EXAMPLE 3

Preparation of (5R? 6s) —5 ~ r [] 2 - fe-hydroxymé'thy, l— pyridinio) ethyl] thio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3 »3« oj hept-2-ene-2-carboxylate 5 OH CH9Om.

C02 ci A. 5-hvdroxymethyl-1- 1 5 1- (2-mercap to ethyl) pyridinium trifluoromethanesulfonate

^ CH20H _ .CH £ OH

25 (/ ON + CFjSOjH + -► HSCH2CHfQ

CF_SO ® D 3 25 Trifluoromethanesulfonic acid (1.327 ml, 0.015 mole) is added dropwise to 3-pyridine-methanol (2.91 ml, 0.030 mole), followed by ethylene sulfide (0.89 ml, 0.015 mole). The resulting homogeneous mixture is heated (oil bath) to 50-70 ° C under nitrogen for 20 hours. The reaction mixture is then taken in water (15 ml) and extracted with CH2Cl2 (5 times 5 ml). The aqueous phase is concentrated in vacuo and then applied to a reverse phase column. Elution with water followed by evaporation of the appropriate fractions gives a pale yellow oil. This material is rechromatographed to give a practically colorless oil. After drying under vacuum (P ^ Q ^) this gives the product (4.50 g, 94%) in the form of a viscous oil. I.R. (film) Ό 3450 (s, OH), 2560 (w, 5 SH) cm-; 1 H NMR (dg-acetone) S: 9.10-8.05 (m, 4H, aromatic), 5.01 (t, J = 5.5 Hz, 2H, N-CH2), 4.93 (s, 2H, -CH20H), 4.43 (br S, 1H, -OH), 3.43-3.18 (m, 2H, S-CH2), 2.34-2.10 (m, 1H, SH) .

j B. p-nitrobenzyl diphenylphosphate (5R, 6S) - 10 3- [2- (3-hydroxymethylpyridinio) ethyl thio ^ -6- [1- (R) -hydroxyethyl ^ -7-oxo-1-azabicyclo Ç5. 2.0] -hept-2-ene-2-carboxylate

, 5 OH OH CH OH

JX> ° - * ^ 0 \ oPNB C02PHB «° ^ ° θ 20 To a solution of p-nitrobenzyl (5R, 6S) -6 - [. 1- (R) -hydroxyethyl] [-3,7-dioxo-1 -azabicyclo [.3.2.0 ^ heptane-2-carboxylate (0.174 g, 0.50 mmol) in 2 ml of anhydrous acetonitrile is added diisopropylethylamine (0.096 ml, 0.55 mmol) at 0 ° C under nitrogen . Diphenyl chlorophosphate (0.114 ml, 0.55 mmol) is then added dropwise and the reaction mixture is stirred at 0 ° C for 30 minutes. A solution of 3-hydroxymethyl-1- (2-mercaptoethyl) pyridinium trifluoromethanesulfonate 30 (0.223 g, 0.70 mmol) in 0.50 ml of acetonitrile is then added, followed by 3 diisopropylethylamine (0.122 ml, 0 , 70 mmol). After standing at 0 ° C for 30 minutes the mixture is concentrated: reaction under vacuum and the residual yellow gum is taken up in water (enough acetonitrile has been added to help dissolve the gum). This solution is applied to a reverse phase column which is eluted with 15% acetonitrile-H ^ O. Lyophilization of the appropriate fractions results in the product (0.305 g, 81%) in the form of a beige solid. I.R. (KBr) Ό · Π13Χ 3420 (br, OH), 1775 (73-lactam CO), 1695 (-CO ^ PNB) cm; 1 H NMR (d ^ -acetone) S: 9.44-7.72 (m, 8H, aromatic), 7.22-6.91 (m, 10H, diphenylphosphate), 5.53, 5.27 (ABq, J = 14 Hz, 2H, benzyl), 5.04 (t, J = 7.4 Hz, 2H, N-CH2), 10 4.75 (s, 2H, CH20H), 4.5-3.1 ( m, 8H), 1.21 (d, J = 6.3 Hz, i 3H, CHMe).

C. (5R, 6s) -3-Ç 2 - (3-hydroxymethylpyridinio) ethyl] - thio ^ C1- (R) -hydroxyethyl-} -7-oxo-1-azabicyclo- t 3.2.0] hept-2- ene-2-carboxylate 15 T / CHz ° H f 9h2 ° h 20 (K \,,, Il _ u ^ Θ C02PNB (0O) 2PO C02ü To a solution of p-nitro-25 benzyl diphenylphosphate (5R, 6S) -3- [2- (3-hydroxymethylpyridinio) ethyl thioj- 6- [1 - (R) -hydroxyé thyl -7-oxo-1 -azabicyclo \ ^ 3.2.0 ~ \ -hept- 2-ene-2-carboxylate (0.145 g, 0.194 mmol) in 10 ml of tetrahydrofuran containing 5 drops of H2O, 6.0 ml of phosphate buffer (0.05 M, pH 7.4), 0.145 g of 10% palladium on carbon are added. carbon and 10 ml of ether The mixture is hydrogenated (by Parr) at a pressure of 275.792KPa s for 1 hour and then filtered through a pad of Celite (trade name). The filter cake X is washed with a little water and ether and the aqueous phase 33 is separated and extracted with ether (3 times). The aqueous solution is then cooled to 0 ° C. and the pH is adjusted to 7.0 with 7.4 pH buffer . After removal of the residual volatile fractions in vacuo the aqueous solution is applied to a reverse phase column of C ^ g 5 which is eluted with water. Lyophilization of the appropriate fractions gives the product (36 mg, 51%) as a slightly yellow solid. Further purification by reverse-phase high-pressure liquid chromatography ("Reverse-phase HPLC") gives the pure product (31 mg, 41%), as a solid. IR (KBr) v> max: 1 3300 (br, OH), 1755 (/ $ - lactam CO), 1590 (-C02) cm ~; 1H NMR (D20): 8.78-7.94 (m, 4H, aromatic), ^, 83 (t, J = 6.0 Hz, 2H, N-CH2), 4.83 (s, 2H, CH20H ), 4'16 (d of q, J = J '= 6.2 Hz, 1H, H-1'), 3.98 (d, of t, 15 £ = 9.1 Hz, = 2.6 Hz , 1H, H-5), 3.75-3.20 (m, 3H), 3.20- 2.65 (m, 2H), 1.22 (d, J = 6.4 Hz, 3H, CHMe ); U.V. (^ 0) XmQX '294 (£ 7614), 266 (£ 6936) nm; t 1 (ph 7.4, 36.8 ° C) 14.0 h.

EXAMPLE 4 20 -

Preparation of (5R, 6s) -3-L 2- (4-hydroxymethylpyridinio) ethylthio1 -6- [, 1- (R) -hydroxyethyl1 -7- oxo-1-azabicyclo C3.2.0J hept-2-ene -2-carboxylate 25

OH

^ θ C02 30 79 A. 4-hydroxymethyl-1- (2-mercaptoethyl) pyridinium 5 trifluoromethanesulfonate 5

^ CH20H h- CF3S03H - * HSCH2CH2 ^ Oy "CH20H

CF, SO-- θ 10 3 3 To a solution of 4-pyridinemethanol (1.635 g, 0.015 mole) in 10 ml of CH2Cl2, at 0 ° C under nitrogen, is added dropwise trifluoromethanesulfonic acid 15 (1.327 ml ; 0015 mole). An oil, yellow-brown separates quickly. An additional equivalent of 4-pyridinemethanol (0.635 g, 0.015 mole) is added to this mixture and the solvent is removed under reduced pressure to give an oil. To this oil is added ethylene sulfide (0.891 ml, 0.015 mole) and the resulting homogeneous mixture is heated (oil bath) at about 60 ° C for 3 hours.

The reaction mixture is then taken in 15 ml of water and the aqueous solution is washed with CHgClg (5 times 5 ml). After removal of the residual organic solvent in vacuo, the aqueous solution is applied to a C ^ g reverse phase column. Elution with water and subsequent evaporation of the appropriate fractions results in an oil which is further dried in vacuo over P20 ^ to give the product (4.64 g, 97%) as an oil. colorless. I.R. (film) \): 3455 (s, OH), 2565 (w, SH) cm ^;

Λ ÏÏlcLX

1 H NMR (dg-acetone) S: 9.07, 8.18 (ABq, J = 6.8 Hz, 4H, aromatic), 5.03 (s, 2H, C ^ OH), 4.96 (t, J = 6.5 Hz, 2H, N-CH2), 4.09 (br s, 1H, -OH), 3.5-3.1 (m, 2H, S-CH2), 2.25 (brs, 1H, -SH).

80 B. P-nitrobenzyl diphenylphosphate (5R, 6S) - 3- [2- (4-hydroxymethylpyridinio) ethyl thioj - 6- £ 1- (R) -hydroxyethyl3 -7-oxo-1-azabicyclo [3.2.oj- hept-2-ene-2-carboxylate 5

OH OH

"VrVo -„ o ^ Un ^ v — 7 o 10 C02PNB C02PNB (îzÎO) aP0 “To a solution of p-nitrobenzyl (5R, 6S) -6- £ l- (R) -15 hydroxyethyl3 -3,7-dioxo -1-azabicyclo [_3.2.J heptane-2-carboxylate (0.348 g, 1.0 mmol) in 5 ml of anhydrous acetonitrile, at 0 ° C. under nitrogen, diisopropylethylamine (0.191 ml, 1 is added dropwise) , 1 mmol) followed by diphenyl chlorophosphate (0.228 ml, 1.1 mmol).

The resulting yellow-gold solution was stirred at 0 ° C for 40 minutes. To this solution is added a solution of 4-hydroxymethyl-1- (2-mercapto-ethyl) pyridinium trifluoromethanesulfonate (0.447 g, 1.4 mmol) in 1 ml of acetonitrile, followed by diisopropylethylamine (0.191 ml , 25 1.1 mmol). A gum-black gum separates from the reaction mixture. After 20 minutes at 0 ° C., the reaction mixture is filtered and concentrated in vacuo. The residue is taken in a minimum volume of acetonitrile-HgO (1: 1) and applied to a C18 reverse phase column. Elution with 25% acetonitrile-H20 and subsequent lyophilization of the appropriate fractions give the product (0.353 g, 47%) as a cream-colored solid. IR (KBr) \) 3240 'ïïlcLX λ - (br, 0H), 1775 (/ ¾ -lactam C0), 1695 (-C02PNB) cm "'; 1 H NMR (dg-acetone) 5: 9,24-7 , 84 (m, 8H, aromatic), 35 7d4-6.9 (m, 10H, diphenylphosphate), 5> 52-5.24 (ABq, 81 J = 14 Hz, 2H, benzyl), 5.15-4 , 80 (m, 4¾ 4.45-3.05 (m, 7H), 1.35 (d, J = 6.6 Hz, 3H, CHMe).

C. (5R, 6S) -3- [2- (4-hydroxymethylpyridinio) ethyl thio] - 6- ^ 1- (R) -hydroxyethyl3 -7-oxo-1-azabicyclo [3.2.0j-5 hept-2- ene-2-carboxylate

OH OH

10 o ^ zô- ^ € ^ H2 ° h C02PNB (^ 0) 2Ρ0Θ C02e 15 A mixture of p-nitrobenzyl diphenylphosphate (5R, 6S) -3- [2- (4-hydroxymethylpyridinio) ethyl thio} -6- £ 1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo ^ 3.2.0 ^ hept-2-ene-2-carboxylate (0.348 g, 0.465 mmol) and 10% palladium on carbon (0.35 g) in 11 ml of phosphate buffer (0.05 M, pH 7.4), 5 ml of THF and 10 ml of ether is hydrogenated at a pressure of 275y792 kPa for 1.25 h.

The mixture is then filtered through a pad of Celite (trade name) and the aqueous phase is washed with ether (3 times). The pH of the aqueous solution is then adjusted to 7.0 using an additional pH 7.4 buffer. After removal of the residual volatile fractions in vacuo the aqueous solution is applied to a C ^ g reverse phase column Elution with 2% acetonitrile-H2O and subsequent lyophilization gives a brown-yellow solid. This material is rechromatographed (reverse phase in C ^ g / ^ O) to give the desired product (Ο, ΟβΟ g, 36%) as a slightly yellow solid.

I.R. (KBr) Ό max: 3400 (br, OH), 1755 (/ 2> -lactam CO), 1590 (~ C02 “) cm-1; 1H NMR (DgO) S: 8.73, 7.96 (ABq, 35 J = 6.8 Hz, 4H, aromatic), 4.93 (s, 2H, CH20H), 82 4.77 (t, J = 6.0 Hz, 2H, N-CHg), 4.15 (d of q, J = J '= 6.3 Hz, 1H, HV), 3.96 (d of t, J = 9.2 Hz, J '= 2.6 Hz, 1H, H-5), 3.65-3.20 (m, 3H), 3.13-2.62 (m, 2H), 1.21 (d, J = 6 , 3 Hz, 3H, CHMe); U.V. (Η £ θ) λ max: 295 (66880), 256 (£ 5595), 5 224 (£ 8111) nm; (pH 7.4, 36.8 ° C) 14.5 h.

EXAMPLE 5

Preparation of 3- [2- (1- (2-methylpyridinium)> - ethylthio ^ -6 <X- [1- (R) -hydroxyethyll -7-oxo-1-10 az abicyclo (3.2.0) hept-2 -ene-2-carboxylate 4

OH

coo "-9 A. 1- (2-Mercaptoethyl) -2- methylpyridinium methanesulfonate

Ms0 ~ L \ '+ NU / + MsOH - * / 25 \ _ / 55 ° C, 21 h To a suspension of 2-methyl-30 pyridinium methanesulfonate in 2-methylpyridine prepared by the addition of acid methanesulfonic acid (0.65 ml, 0.010 mole) to cold 2-methylpyridine (2.17 ml, 0.022 mole) ethylene sulfide (0.655 ml, 0.011 mole) is added.

The reaction mixture is stirred under a nitrogen atmosphere at 55 ° C for 21 hours, cooled to 23 ° C and diluted 83 with water (5 ml). The aqueous solution is washed with ether (6 times 4 ml), pumped to remove traces of organic solvents and poured onto the top of a column (2.5 x 10.0 cm) of micro-bondapak C- 18 (trade name 5). The column is eluted with water and the lyophilization of the appropriate fractions gives 2.13 g (85%) of the title compound, IR (film) 0 ω & χ: 2520 (SH), 1623 (pyridinium) 1574, 1512, 1485 , 1412, 1195 (sulfonate), 1038 cm "1; 1H NMR (DMSO-dg + D20) <5: 2.37 (3H, s, CH ^ SO ^), 10 2.83 (3H, s, CH ^ on pyridinium), 3.09 (2H, J = 6.9 Hz, CH2S), 4.71 (2H, t, J = 6.9 Hz, 0Η £ Ν +), 7.93 (2H, m, Hm pyridinium), 8.44 (1H, m, pyridinium Hp), 8.89 (1H, m,

Ho de pyridinium), U.V. (H20) ^ max: 266 ((.3550) m ^ tu.

15 B. Paranitrobenzyl diphenylphosphate 3- £ 2- 1- (2-methylpyridinium)) ethylthio 3 -6 <X - \ _1- (R) ~ hydroxyethyl] -7-oxo-1-azabicyclo (3 »2.0) hept- 2- ene-2-carboxylate 20

1) NEt (iPr) p nM

OH 0 f 25 0> 3) © / Ms0 0 0 COOPNB / - COOPNB (ph0) ^ (j · 4) NEt (iPr) 2 30 To a cold solution (0 ° C) of p-nitrobenzyle 6o (- j ^ 1- (R) -hydroxyethyl3 -3,7-dioxo-1 -az abicycio (3 · 2.0) -heptane-2-carboxylate (0.523 g, 1.50 mmol) in acetonitrile (6 ml) maintained under an atmosphere diisopropylethylamine (0.314 ml, 1.80 mmol) is added with nitrogen followed by diphenyl chlorophosphate (0.373 ml, 1.80 mmol), the reaction mixture is stirred for 30 minutes at 0 ° C. and treated with a solution of 1- (2-mercaptoethyl) -2-methylethylpyridinium methanesulfonate (0.530 g, 2.16 mmol) in acetonitrile (18 ml) followed by diisopropylethylamine (0.314 ml, 1.8 mmol). The reaction mixture is stirred at 0 ° C for 1 hour, diluted with cold water (0 ° C) (26 ml) and poured onto the top of a micro column (3.5 x 7.0 cm) -bondapak C-18 10 (trade name) Eluting the column with 2596 acetonitrile-75% water and with 50% acetonitrile-s 50% water gives apr upon lyophilization of the appropriate fractions 1.06 g, (96%) of the title compound in the form of a yellowish powder, IR (KBr) O: 3650-3100 (0H), 1770 (C = 0 15 dey?> - lactam), 1693 and 1690 (C = 0 of the PNB ester), 1630 (pyridinium), 1595 (phenyl), 1518 (NO2), 1335 (NO2), 890 cm “1 (NO2), 1 H NMR (DMSO , dß) S: 1.15 (3H, d, J = 6.1 Hz, CH ^ CHOH), 2.87 (s, CH ^ on pyridinium), 3.6-4.4 (2H, m, H -5, CH3CH0H), 4.75 (2H, m, CH ^ N4), 5.37 (center of ABq, 20 J = 14 Hz, CH2 of PNB), 6.5-7.4 (10H, m, phenyl), 7.70 (2H, d, J = 8.8 Hz, Ho of PNB), 8.0 (2H, m, Hm of pyridinium), 8.24 (2H, d, J = 8.8 Hz , Hm of PNB), 8.50 (1H, m, Hp of pyridinium), 8.95 (1H, brd, J = 6.1 Hz, Ho of pyridinium), UV (H20) Amax: 265 (611990), 314 (£ 8020) mjt ,.

25 C. 3- Γ2- (1- (2-methylpyridinium)) ethylthio1 -6 <Y- L 1- (R) -hydroxyethyl] -7 — oxo-1-azabicyclo (3.2.0) -hent-2-ene -2-carboxylate

30 OH OH

COOPNB (PhO) DP0 C00: 35 * 85 To a solution of p-nitro-benzyl diphenylphosphate 3- £. 2- (1- (2-methylpyridinium)) ethylthio ~ \ -6χ- jjl- (H) -hydroxyethyl ^ -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate (0, 66 g, 0.90 mraole) in wet tetrahydrofuran (34 ml), ether (34 ml), monobasic potassium phosphate-sodium hydroxide buffer (0.15 M, 16.5 ml, are added). pH 7.22) and 10% palladium on charcoal (0.66 g). The resulting mixture is hydrogenated under a pressure of 275.792 kPa at 23 ° C for 1.25 hours.

The organic layer is separated and extracted with buffer (twice 6 ml). The aqueous layers are combined, filtered through a pad of Celite (trade name), washed with ether (40 ml), pumped to remove traces of organic solvents and poured on top of a 15 column (2.5 x 10 cm) of micro-bondapak C-18. Elution from the column with water and lyophilization of the appropriate fractions give the title compound (0.098 g, 31%) in the form of a yellowish powder, IR (KBr) v: 3650-3100 (OH), 1755 (0 = 0 de / ½-lactarne), 1630 (pyridinium), 1595 cm ”20 (carboxylate), 1H NMR (DgO) 1.20 (3H, d, J = 6.3 Hz, CH3CH0H), 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, CH ^ HOH), 4.78 (t, J = 6.2 Hz, CH2N), 7 , 8 (2H, m, Hm of pyridinium), 8.3 (1H, m, 25 Hp of pyridinium), 8.65 (1H, m, Ho of pyridinium), 23 UV (H20) 'Xmax: 268 (fc9350 ), 296 fe8840) m ^, [(¾¾ + 41 ° (c 0.5, H2O), t ^ = 15.0 h (measured at a concentration of 10 ”4 M in phosphate buffer pH 7.4 at 36.8 ° C).

86 EXAMPLE 6

Preparation of 3- t2- (1- (4-methylpyridinium)) - ethylthio -6o <- 1 (R) -hydroxyethylJ -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate 5 râ- 10 q / ^ coo "A. Methanesulfonate of 1- (2-mercaptoethvl) -4- mé thvlpvridinium 15 â \ + + Ms0H 55 ° C, * 1150 '-' 24 h '20 A suspension of methanesulfonate 4 -picolinium in 4-picoline prepared by the addition of methanesulfonic acid (0.65 ml, 0.010 mole) to 4-picoline (2.14 ml, 0.022 mole) while cooling, sulfide d ethylene (0.655 ml, 0.011 mole) The reaction mixture is stirred under an atmosphere of nitrogen at 55 ° C for 24 hours, cooled to -23 ° C and diluted with water (5 ml) and ether (10 ml), separate the organic layer and wash the aqueous layer with ether (5 x 5 ml) and apply to the top of a micro column (2.5 x 10 cm) -bondapak C-18 after the traces of ether have been removed under reduced pressure Elution from the column with a mixture of 15% acetonitrile ~ -85% water after lyophilization gives the appropriate fractions of a colorless syrup 2.66 g (100%), IR (film) S) max: 87 2500 (SH), 1640 (pyridinium), 1572, 1520, 1478, 1200 ( sulfonate), 1040, 833 and 768 cm “1, 1H NMR (DMSO-dg) S: 2.31 (3H, s, CH ^ SO ^), 2.62 (s, CH ^ on pyridinium), 2.2 -2.9 (4H, SH, CH ^ on pyridinium), 3.04 (2H, m, CH2S), 5 4.68 (2H, t, J = 6.4 Hz, CH2N +), 8.01 (2H , d, J = 6.6 Hz, Hm of pyridinium), 8.89 (2H, d, J = 6.6 Hz, Ho of pyridinium), UV (H20) Amax: 256 (64100), 221 (67544) m ^.

B. 1- (2-mercaptoethyl) -4- methylpyridinium / S \ + + pTsOH - "H ^ © PTS ° 15 p-toluene sulfonate To a suspension of p-toluenesulfonic acid (1.72 g, 0.01 mole) in benzene (6.5 ml) 4-picoline (1.17 ml, 0.012 ml) is added. The resulting mixture is stirred under a nitrogen atmosphere at 23 ° C for 30 minutes, treated with ethylene sulfide (0.65 ml, 0.011 mole) and stirred at 75 ° C for 24 hours. More ethylene sulfide (0.65 ml, 0.011 mole) was added and stirring continued at 75 ° C for an additional 24 hours. The reaction mixture is cooled to 23 ° C and diluted with water (5 ml) and ether (8 ml). The aqueous layer is separated and washed with ether (3 times 8 ml). The traces of organic solvents are removed in vacuo and the compound 30 is chromatographed on micro-bondapak C-18 and water as elution solvent to give 2.94 g (90%) of the title compound as a colorless syrup; IR (film) \) max: 2510 (SH) 1640 (pyridinium), 1595, 1582, 1475, 1200 (sulfonate), 1031, 1010, 818 cm “1, 1H NMR (DMSO, dg) 5: 2.29 ( 3H, 35 s, CH ^ on pyridinium), 2.61 (s, CH · ^ Ph), 2.4-2.8 (4H, SH, 88 CH ^ Ph), 3.03 (2H, m \ addition of D ^ O 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 = 7.9 Hz, phenyl), 8.00 (2H, d, J = 6.5 Hz, Hm of pyridinium), 8.89 (2H, d, J = 6.5 Hz, Ho pyridinium), 5 UY (H20) X max: 256 (£ 4,315), 222 (£ 17,045) ψ * ..

C. Paranitrobenzyl diphenylphosphate 3—2— (1 - (4-methylpyridiniuin)) ethylthio -6ςχ - _1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-10 ene- 2-carboxylate

D NEt (iPr) 2 QH

Oh y

€ >> '0 · ° (M

4) NEt (iPr) 2 20 In a cold solution (0 ° C) of p-nitrobenzyl 6 <X _ L1- (R) -hydroxyethyl ^ \ -3,7-dioxo-1-azabicyclo (3.2û) heptane- 2-carboxylate (0.522, 1.5 mmol) in acetonitrile (6 ml) maintained under a nitrogen atmosphere, diisopropylethylamine (0.314 ml, 1.8 mmol) is added followed by diphenyl chlorophosphate (0.373 ml, 1.9 mmol). The reaction mixture is stirred for 45 minutes and treated dropwise with a solution of 1- (2-mercaptoethyl) -4-methylpyridinium methanesulfonate (0.559g, 2.16 mmcJes) in acetonitrile (1.8 ml ) followed by diisopropylethylamine (0.314 ml, 1.8 mmol). The mixture is stirred: reaction at 0 ° C for 1 hour, diluted with cold water (0 ° C) (24 ml) and poured at the top of a column (2.5 x 8.5 cm) of micro-bondapak C-18. Elution from the column first with a mixture of 259% acetonitrile-75% water (100 ml) then with a mixture of 50% acetonitrile-50% water ( 100 ml) produced after lyophilization of the appropriate fractions 0.91 g (83%) of the title compound in the form of a yellowish powder, IR (KBr) max: 3700-5 2800 (0H), 1770 (C = 0 of / Ulactam), 1700 (C = 0 of the PNB ester), 1640 (pyridinium), 1595 (phenyl), 1520 (N02), 1340 (N02), 890 cm "1 (N02), 1 H NMR (DMSO, dg) S: 1.16 (3H, d, J = 6.2 Hz, CH- ^ CHOH), 2.61 (s, CH ^ on pyridinium), 3.1-3.7 ^ (3H, m, H- 6, CH2S), 3.7-4.4 (2H, m, H-5, CH ^ CHOH), 4.79 (2H, 10 brt, J = 6.3 Hz, CH2N +), 5.17 (d , J = 4.9 Hz, OH), 5.37 (center of ABq, J = 14.1 Hz, CH2 of PNB), 6.7-7.4 (10 H, m, phenyl), 7.69 (2H, d, J = 8.8 Hz, Ho of PNB), 8.00 (2H, d, J = 6.5 Hz,

Hm of pyridinium), 8.23 (2H, d, J = 8.8 Hz, Hm of GNP), 8.92 (2H, d, J = 6.5 Hz, Ho of pyridinium), UV (H „0 ) λ: 15 262 ((: 10835), 311 ((.9670) m ^ e. Anal, hold, for C36H36N3 ° 10SP * 1'5 H20: 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.

D. 3- [.2- (1- (4-methylpyridinium)) ethylthio ^ -6o (- 20 L1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) - hept-2- ene-2-carboxylate

OH OH

* - 10% Pd / C, W

25 lO-s ~ ^ 9 vq-s ^^ (Ph0) Jo- tampon ^ U N 2 C00 “

COOPNB

30 To a solution of p-nitrobenzyl diphenylphosjhate of 3- £ - (1- (4-methylpyridinium)) -ethylthio "i -6 ° (- Li- (R)“ hydroxyethyl] - 7-oxo-1-azabicyclo ( 3.2.0) hept-2-ene-2-carboxylate (0.587 g, 0.80 mmol) in wet tetrahydrofuran (30 ml) ether (30 ml), monobasic potassium phosphate buffer 90 are added -sodium hydroxide (0.15 M, 14.7 cil, pH 7.22) and 10% palladium on charcoal (0.59 g).

The resulting mixture is hydrogenated under a pressure of 275.792 kPa at 23 ° C for 1.25 hours. The organic layer is separated and extracted with the buffer (twice 6 ml).

The aqueous extracts are combined, filtered through a pad of Celite (trade name), washed with ether (5 times 20 ml), pumped to remove traces of organic solvents and poured on top of '' a column 10 (2.5 x 10 cm) of micro-bondapak C-18 (trade name). Elution from the column with water and lyophilization of the appropriate fractions gives 0.136 g (49%) of the title compound as a yellowish powder; IR (KBr) "0 max: 3700-3000 (OH), 1770 (C = 0 de / i-lactam), 15 1642 (pyridinium) 1592 cm-1 (carboxylate), NMR (D20): 1.19 (3H , t, J = 6.3 Hz, CH ^ CHOH), 2.59 (3H, s, CH3 on pyridinium), 2.84 (d, J = 9.1 Hz, H-4), 2.90 ( d, J = 9.1 Hz, H-4), 3.0-3.6 (3H, m, CH2S, H-6), 3.86 (dd, J = 9.1 Hz, J = 2, 6 Hz, H-5), 4.12 (m, CH ^ CHOH), 4.5-4.9 (CH ^ N4 "masked by HOD), 20 7.80 (2H, d, J = 6.6 Hz, Hm of pyridinium), 8.58 (2H, d, J = 6.6 Hz, Ho of pyridinium), UV (H20) Λ max: 256 (¢ 5510), 262 (65360), 296 (£ 7050) myuu, [p <\ Z3 + 20.8 ° (C 0.48, HgO), tj ,. = 12.8 h (measured at a concentration of 10 ”^ M in a

S

phosphate buffer pH 7.4 at 36.8 ° C).

25 ’EXAMPLE 7

Preparation of (5R) 3 ~ C2- (4-methylthiopyridinio) -ethylthio-] - (6S) -Q (1R) -hydroxyethyl3 -7-oxo-1-azabicyclo |] 3.2. θ '} hept-2-ene-2-carboxylate

30 OH

GOf 91 y.

A. 4-methylthiopyridine

c / P = S \ 1) Mel / EtOH

N ^ / “SH - * Ni) VsMe '-' 2) NaOH '-' ^ Dissolve 4-mercaptopyridine (5.55 g, 50.0 mmciles · 10 Aldrich) in boiling absolute ethanol (50 ml). The. insoluble material is removed by filtration on Celite.

The filtrate is heated to redissolve and then it is cooled to about 50 ° C, methyl iodide is added all at once (3.17 ml, 51.0 mmol; Aldrich). The mixture is cooled to crystallize. Filtration of the solid gives 6.77 g (26.7 mmol, yield 53.550 of the title compound in the form of hydrogen iodide: 1 H NMR (D20) S: 2.70 (3 H, s, -SCH3) and 7 , 65-7.77-0.35- 8.48 ppm (4H, type A ^^, aromatic-H s); IR (Nujol) Omax: 20 1615, 1585 (aromatic) and 780 cnT'X UV (Ho0) X : 227 (£ 2.02 x 10H) and 298 nm (£ 1.64 x 10 ^).

The iodide (6.33 g, 25.0 mmol) is dissolved in H2O (40 ml) and the insoluble material is removed and washed with H2O (10 ml). To the filtrate, NaOH in 25 granules (5 g) is added at 0-5 ° C. and extracted with Et 2 O (3 times 25 ml), saturating the aqueous layer with NaCl. The combined organic extracts are washed with salt water (x 2), dried (MgSO 4) and evaporated, obtaining 2.92 g (23.4 mmol, total yield 50%) of the title compound as oil: 1 H NMR (CDCl3) S: 2.48 (3H, s, -SCH3) and 7.03-7.13-8.38- 8.48 ppm (4H, type A9B9, aromatic-HS); I.R. (film) ^: 1580 and 800 cm "1.

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

92 B. 4-methylthio-N- (2-mercaptoethvl) pyridinium methanesulfonate> v · 3 3

4-Methylthiopyridine (2.75 g, 22.0 mmol) is added slowly to methanesulfonic acid (0.65 ml, 10.5 mmol) while cooling in an ice bath. To this solid is added ethylene sulfide (0.66 ml, 11.0 mmd Aldrich) and the mixture is heated to 50-60 ° C for 21 hours. As the reaction takes place the solid passes 15 solution. After cooling, the reaction mixture is dissolved in (5 ml) and washed with Et 4 O (5 times 4 ml).

The cloudy aqueous layer is filtered through Celite and the filtrate is purified by chromatography on a column of reverse phase silica gel (microbondapak C-18 10 g), eluting with water. Each 10 ml fraction is collected. Fractions 2 and 3 are combined and repurified by the column in reverse phase. Fraction 2 gives 1.258 g (4.48 mmol, yield = 42.6%) of the title compound in the form of a viscous oil: 1 H NMR (DMSO-dg, CFT-20) S: 2.32 (3 H , s, MeS03G), 2.72 (3H, s, -SMe), 2.68 (1H, m, SH), 2.9-3.2- (3H, m, -CHgS-), 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, aromatic-HS); I.R. (pure) ί> w: 1630, 1200 (br, Θ λ max), 785 and 770 cm Ί.

D Ή · 30 These reagents are distilled before use.

93 C. (5R) p-nitrobenzyl chloride 3-L 2- (4-methyl-thiopyridinio) ethylthio j - (6s) - \ (iR) -hydroxyethyl - 7-oxo-1-azabicyclo C3.2.01 hept-2 -ene-2-carboxylate

5 OH OH

/ "" T d.

10 “o ™“ ο "“ To a solution of (5R) p-nitrobenzyl 3,7-dioxo- (6S) ~ (iR) -hydroxyethyl] -1-azabicyclo [3.2.0 ^ heptane-15 (2R) - carboxylate (475 mg, 1.36 mmol) and diisopropylethylamine (0.24 ml, 1.4 mmol) in CH ^ CN (5 ml), chlorophosphate is added at 0 ° -5 ° C under nitrogen of diphenyl (0.29 ml, 1.41 mmol) The mixture is stirred at 0 ° -5 ° C. for 30 minutes to which is added an oily suspension of 4-methylthio-N- (2-methanesulfonate) -mercaptoethyl) pyridinium (678 mg, 1.45 mmol; 60% pure) in CH ^ Cl (1.5 ml) followed by diisopropylethylamine (0.24 ml, 1.4 mmol). The mixture is stirred at 0 ° -5 ° C for w 1 hour Immediately after addition of the base, a yellowish precipitate is formed The precipitate is filtered and washed with cold CH 3 CN (3 ml), obtaining 413 mg of yellowish solid. -this is triturated from 10% MeOH in H2O (5 ml) to obtain 341 mg (0.618 mmol, yield = 45.4%) of the title compound in the form of 30 white crystals: PF 11 8 ° -120 ° C; 1 H NMR (DMSO-dg, CFT-20) S: 1.16 (3H, d, J = 6.1 Hz, l'-CH ^, 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, -C ^ N®), 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, 94 nitrophenyl) -HS) 7.97 (2H, "d", J = 7.0 Hz, pyridinio-HS), 8.25 (2H, "d", J = 8.8 Hz, nitrophenyl -Hs), and 8.76 ppm (2H, "d", J = 7.1 Hz, pyridinio-Hs); I.R. (Nujol) Ό: 3250 (OH), 1775 (fî-lactam), 1700 (ester) and 1625 cm “(pyridinio); 5 U.V. (abs, Et OH) λ: 308 nm (£ 4.47 x 104); T ^] n25 ° + 24.8 (c 0.5, MeOH); Anal, hold, for C2ZfH26N306S2Cl.H20: C 50.56, H 4.95, N 7.37; found: C 50.63, H 4.72, N 6.89.

10 D. (5R) 3- [2- (4-methylthiopyridinio) ethylthio3 - (6s) - (IR) -hydroxyethyl ~ [-7-oxo-l-azabicyclo L 5.2.θ1 -hept-2-ene-2- carboxylate

15 OH OH

V ...... .‘ίδ / ™ · 7- · 'Vr ......

20 C02PNB C02

(5R) p-nitrobenzyl chloride 3 ~ L2- (4-methyl-thiopyridinio) ethylthio 3 - (6S) - ^ (1 R) -hydroxyethyl3 -7-oxo-1-azabicyclo b »2.ol hept-2 -ene-2-carboxylate (380 g, * r 25 0.688 mmol) is dissolved in THF (31.5 ml) and phosphate buffer at pH 7.40 (31.5 ml; 0.05 M Fisher) and diluted with Et20 (31.5 ml). This solution is mixed with 10% -C palladium (380 mg, Engelhard) and hydrogenated at a pressure of 241.318 kPa on the Parr shaker at room temperature for 1 hour. The aqueous layer is filtered through Celite to remove the catalyst and the Celite pad is washed with H2O (2 times 5 ml). The filtrate and the washes are combined and washed with Et20 (2 times 30 ml).

* The aqueous layer is pumped to remove all organic solvents and purified by reverse phase column chromatography (microbondapak C18, 13 g, trade name of Waters Associates), eluting with E ^ O. The fractions having a UV absorption at 307 nm are collected (approximately 1 liter) and lyophilized to obtain 127 mg 5 (0.334 mmol, yield = 48.590 of the title compound in the form of a yellowish powder: NMR (D20, CFT-20) 5: 1.20 (3H, d, J = 6.4 Hz, 1'-CH3), 2.64 (3H, s, -SCHj), 2.81 (2H, m, -SCi £ -), 3 , 19 (1H, dd, ^, = 6.1 Hz, J6_5 = 2.6 Hz, 6-H), 3.32 (2H, dd, J = 11 Hz, J-5.5 Hz, 4-Hs ), 3.92 (1H, dt, 10 J = 9.2 Hz, J3__6 = 2.6 Hz, 5-H), 4.1 (1H, m, 1'-H), 4.61 (2H, t, J = 5.9 Hz, -CHgN®), 7.70 (2H, "d”, J = 7.1 Hz, aromatic-Hs), and 8.40 ppm (2H, "d", J = 7.1 Hz, aromatic-Hs); IR (KBr, disc) -0 max: 3400 (OH), 1750 (ß-lactam), 1630 (pyridi-nium) and 1590 cirf ^ (carboxylate); UV (Ho0) Λ: 231 15 (29800) and 307 nm (£ 25000); + 3.14 (ç 0.5, H2O).

EXAMPLE 8

Preparation of 3-1 2- (3-methoxy-1 -pyridinium) -ethylthio ^ -6o6 - (j '- (R) -hydroxyethyl 1 -7-oxo-1-20 azabicyclo (3.2.0) -hept-2 -ene-2-carboxylate

OH

T

_ OMe 25 C02e A. 1- (2-Mercaptoethyl) -3- methoxypyridinium 30 MeO methanesulfonate _.__ 0

\ MSO

“- <0) * Δ JÏ2L- 96 A of precooled 3-methoxypyridine (5 ° C) (698 mg, 6.6 mmol) methanesulfonic acid (0.216 ml, 3.05 mmol) is added dropwise ethylene sulfide (0.19 ml, 3.2 mmol) The mixture is then heated at 60 ° C for 18 hours, cooled to 20 ° C, diluted with water (10 ml) and washed with ether (3 times 10 ml) The aqueous phase is pumped under high vacuum for 15 minutes and poured onto a C ^ g reverse phase column.

"The title compound is eluted with water. The appropriate fractions are combined and evaporated under high vacuum: to give the desired thiol (61.6 mg, yield 76.3%); IR (CH9C19) O: 2550 (w, SH) and 1620, 1600, 1585 cm ”1 (m, aromatic); NMR H (DMSO d ^) o: 8.90-7.90 (4H, m, aromatic CH), 4.72 (2H , t, J = 6.6 Hz, C ^ N *), 4.01 (3H, 15 s, OCH ^), 3.5-3.0 (m, CH ^ S masked), 2.66 (1H , dd, J = 9.5 Hz, J = 7.5 Hz, SH) and 2.31 ppm (3H, s, CH ^ SO ^).

B. para-nitrobenzyle 3 Ç2- (3-methoxy-1-pyridinium chloride) ethylthio3 -6û <-L 1 '- (R) -hydroxyéthylJ -20 7-oxo-1-azabicyclo (3.2.0) hept-2- ene-2-carboxylate

OH 1) EtN (- <) 9 nH

f v 2. niG OMe

L. - 2) C1PO (0 ?!) T C1 J

25 tlVo _Λ / Γ-τΛ ® / 7 = Λ ο ^ ~ Νγ 3) hs ^ (o ^ ;; o-

C0? GNP

4) EtH (_Oz 2 30

A cooled (0 ° C) solution of p-nitrobenzyl 6 <X-C1 '- (R) -hydroxyethyl3 -3,7-dioxo-1-azabicyclo- (3.2.0) -heptane-2-carboxylate (1.04 g, 3 mmol) in acetonitrile (12 ml) is treated dropwise with diisopropylethylamine (0.63 ml, 3.6 mmol) and diphenyl chlorophosphate (0.75 ml, 3, 6 mmol) and stirred at 0 ° C for 30 minutes. The resulting enol phosphate is treated with 1- (2-mercaptoethyl) -3-methoxypyridinium methanesulfonate (1.14 g, 4.30 mmol) in CH ^ CN (7 ml), diisopropylethylamine (0.63 ml, 4.30 mmol), stirred for 30 minutes and cooled to -10 ° C for 30 minutes.

The solid which precipitates from the mixture is filtered, washed with cold acetonitrile (2 ml) and dried to give the title compound (1.32 g, yield 82%); IR (Nujol) ν ': 10 3320 (m, OH), 1780, 1765 (s, / 4 -lactam C = 0), 1700, 1695 (m, ester C = 0) and 1520 cm ^ (s, NO2) > * NMR (DMSO dg): 9.01 (1H, bs, H-3 aromatic), 8.75 (1H, bd, J = 5.4 Hz, H-6 aromatic), 8.35-7.95 (4H, m, H-aromatic), 7.70 (2H, d, J = 7.7 Hz, H-aromatic), 5.37 (2H, center of ABq, 15 J = 13 Hz, CH2PNB), 5 , 17 (1H, d, J = 4.9 Hz, OH), 4.87 (2H, t, J = 6.3 Hz, CH2-N +), 4.35-3.75 (2H, m, H -5 and H-1 ·), 4.00 (3H, s, OCHj), 3.56 (part of a 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, CH ^ CHO).

20 C. 3- ^ 2- {3-methoxy-1-pyridinium) ethylthio 3 -6 (X- y 1 '- (R) -hydroxyethyl3 -7-oxo-1-azabicyclo (3.2.0) -hept-2 -ene-2-carboxylate OH o OMe OH ° Me '25 l Cl0 / _ /] -r>, ¾¾ VV® A * - / ^ 0Α'γ J0 CO-jPNB CO ©

A solution of para-nitrobenzyl 3 12- (3-methoxy-1-pyridinium chloride) ethylthio -6- 111- (R) -hydroxyethyl -35 7-oxo-1-azabicyclo (3.2.0) hept-2-ene 2-carboxylate (600 mg,!! 98 1.12 mmol) in THF (25 ml), ether (25 ml) and phosphate buffer at pH 7.4 (0.1M, 25 ml) is hydrogenated in a Parr shaker on 10% palladium / C (1.1 g) for 1 hour at a pressure of 275.792 kPa. The mixture is diluted with ether and the aqueous phase is filtered through a hardened filter paper equal to ^ 54. The aqueous layer is washed with ether (2 times 20 ml), pumped under vacuum and poured onto a reverse phase column of silica gel 3. 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 repurified by HPLC to give the penem carboxylate (150 mg, 38%); I.R. (Nujol): 1750 (s, Ä-lactarne C = 0) mm Λ ^ A -.

and 1580 cm “(s, carboxylate); NMR H (D20) o: 8.55-8.30 15 (2H, m, H-2, H-6 aromatic), 8.17-7.75 (2H, m, H-3, H-4 aromatic ), 4.47 (2H, t, J = 5.9 Hz, CH2î ^), 4.10 (1H, part of 5 lines, J = 6.3 Hz, H-1 '), 3.97 (3H , s, OCH ^), 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, 20 J = 2.6 Hz, H-6), 2.99-2.60 (2H, 6 lines, part of H -4), and 1.20 ppm (3H, d, J = 6.4 Hz, CH3); U.V. (H £ 0, c 0.05) λ max: 290 (£ 10517), 223 (£ 6643); t ^ (0.1 M pH 7.4 phosphate buffer, 37 ° C) 20 h.

EXAMPLE 9 Preparation of (5R, 6s) -3- [2- (3-methylthiopyridinio) -ethyl thio 1 -6- Z1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0) hept-2-ene-2-carboxylate OH SMe * JVr- \ JVo> oJ-N ^ S 'CO θ 55 2 99 A. 3-Methlthio-1- (2-mercaptoethyl) chloride - pyridinium SMe 5 ® / - <^ + HCl + ^ - »HSCH2CH2Ng> Cl® 10 To a solution of 3-methylthiopyridine (2.00 g,

0.016 mol) in 10 ml of ether is added 15 ml of 1N HCl

and the mixture is then shaken. The aqueous phase is separated with 10 ml of ether and then evaporated. The residual hydrogen chloride is then dried under vacuum (P ^ O ^) to give a white solid. To this hydrochloride solid, 3-methylthiopyridine (1.88 g, 0.015 mole) and ethylene sulfide (0.89 ml, 0.015 mole) are added and the resulting mixture is heated (oil bath) at 55-65 ° C under nitrogen for 15 hours. This gives a slightly turbid oil which is taken in 125 ml of water and washed with CH2Cl2.

The aqueous solution is concentrated to approximately 25 ml and then a few drops of acetonitrile are added to make the mixture homogeneous. The resulting aqueous solution is applied to a C18 e reverse phase column. Elution with water and subsequent evaporation of the appropriate fractions results in the product (2.66 g, $ 80) as a viscous oil. pale yellow. I.R. (film) Ό: 2410 a λ ¢. max (br, -SH) cm “; H NMR (dg-DMSO + D ^) to: 8.88-7.88 (m, 4H, aromatic), 4.70 (t, J = 6.5 Hz, 2H, N-CH2), 3.08 (bias t, 30 J = 6.5 Hz, 2H, S-CHj, 2.64 (ε, 3H, S-Me).

“Ί,,

Prepared by the method of J.A. Zoltewiez and C. Nisi, J. Org. Chem. 34, 765 (1969).

100 B. p-nitrobenzyl chloride (5R, 6S) -3- LM3- methylthiopyridinio) ethyl thio - 6-ti- (R) -hydroxyethyl3 -7-oxo-1-azabicyclo £ 3.2.03 hept- 2-ene- 2-carboxylate 5 QPj OH Sl-ie 'Vr>. - “s C02PNB co2pnb

A solution of p-nitrobenzyl (5R, 6S) -6- i— (R) - 15 hydroxyethyl J -3,7-dioxo-1-azabicyclo (3.2.0) heptane-2-oarboxylate (0.522 g, 1.50 mmol) in 7 ml of anhydrous acetonitrile is cooled to 0 ° C. and then diisopropylethylamine (0.287 ml, 1.65 mmol) is added dropwise.

To the resulting yellow-brown solution is added dropwise diphenyl chlorophosphate (0.342 ml, 1.65 mmol) and the reaction mixture is kept at 0 ° C for 30 minutes. Diisopropylethylamine (0.313 ml, 1.80 mmol) is then added, followed by a solution of 3-methylthio-1- (2-mercaptoethyl) pyridinium 23 chloride (0.398 g, 1.80 mmol) in 0.70 ml of anhydrous DMF. About a minute after the addition is complete, a precipitate separates from the reaction mixture and additional cooling at -10 ° C for 10 minutes gives a solid mass of orange color. This solid is subsequently triturated with acetonitrile and the residue is collected by filtration. The residue is washed with acetonitrile, then acetone and finally dried in vacuo to give the product (0.455 g, 55%) as a cream-colored solid. The combined filtrate is evaporated to give a yellow oil which is taken up in a minimum volume of acetonitrile and cooled to 0 ° C for 30 minutes. Filtration of this mixture produced an additional 0.139 g of the product as a light yellow solid. The combined yield is 0.594 g (7296). I.R. (KBr) Ό 3345 (br, 5 -OH), 1770 (fl -lactam CO), 1680 (-C02PNB) cm “n; NMR (dg-DMSO) S; 8.98-7.96 (m, 4H, aromatic pyridinium) ^ 8.20-7.65 (ABq, J = 7.0 Hz, 4H, aromatic GNP), 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).

* 10 c. (5R, 6s) -3-t2- (3-methylthiopyridinio) ethyl thio3 ~ 6- Ll- (R) -hydroxyethyl ^ -7-oxo-1-azabicyclo Χ, 3.2.θ] -hept-2-ene-2 -carboxylate 15 OH SMe OH gMe V> ^ - * 20 Wd® co2pnb co2g A mixture of p-nitrobenzyl chloride (5R, 6S) -25 3- - (3-methylthiopyridinio) ethyl thio3 -6- £ l- ( R) - hydroxyethylj -7-oxo-1-azabicyclo £ 3.2.θΊ ^ hept-2-ene-2-carboxylate (0.551 g, 1.0 mmol) and 10% palladium on carbon (0.55 g) 5 ml of THF and 25 ml of ether are added to 25 ml of phosphate buffer (0.05 M, pH 7.4).

This mixture is hydrogenated (Parr) at a pressure of 275,792 KPa for 1 hour. The reaction mixture is then filtered through a Celite pad and the filter cake is washed with H2O and ether. The aqueous phase is separated and washed with additional ether (3 times).

102

After removal of the residual organic solvents under vacuum, the aqueous solution is cooled to 0 ° C. and the pH is adjusted to 7.0 with saturated aqueous NaHCO 4. This solution is immediately applied to a reverse phase column 5 in C ^ g. Elution with water and subsequent lyophilization of the appropriate fractions produces 0.25 g of a bright yellow solid. This material is repurified by reverse phase HPLC to give the product (0.210 g, 55%) in the form of a light yellow solid. I.R. (KBr) Ό HIcLX. 'Μ - 10 5400 (br, -OH), 1755 (y2> -lactam CO), 1590 (-CO ^ -) cm; 1 H NMR (Dg0) S: 8.60-7.76 (m-4H, aromatic), 4.76 (t, J = 5.8 Hz, 2H, N-CH2), 4.13 (d of q) , J = J '= 6.3 Hz, 1H, H-1'), 3.95 (d of t, J = 9.0 Hz, J '= 2.8 Hz, 1H, H-5), 3 , 45-2.75 (m, 5H), 2.59 (s, 3H, S-Me), 1.20 (d, J = 6.4 Hz, 3H, CHMe); 15 U.V. (HgO) Λ max: 296 ¢ 68509), 273 (£ 13005), 231 (€ .11576) nm; ti (pH 7.4, 36.8 ° C) 20 h.

EXAMPLE 10

Preparation of 3-L2- (1- (2,6-dimethylpyridinium) -ethylthiol -6c * - [] l- (R) -hydroxyethyll -7-oxo-1-20 azabicyclo (3 ♦ 2.0) hept-2-ene -2-carboxylate OH CH3 coo "ch3 A. 1- (2-Mercaptoethyl) methanesulfonate ~ 2? 6r dimé thylpyridinium 30 1 + Ms OH -► V / 100 ° C, 42 h MsO” 35 103

A mixture of 2,6-dimethylpyridine (19.2 ml, 0.165 mol) and methanesulfonic acid (3.27 ml,

0.050 mole) for 15 minutes, treat with ethylene sulfide (4.17 ml, 0.070 mole) and stir at 100 ° C

5 for 42 hours under a nitrogen atmosphere. 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 organic layer is extracted with water (2 x 5 ml). The aqueous layers are combined, ώ ίο filtered through a pad of Celite, washed with ether (2 times 15 ml), pumped to remove traces of organic solvents and poured at the top of a column (3, 0 x 12 cm) of micro-bondapak C-18 (trade name). Elution with a 3% mixture of acetonitrile-15,979 ^ of water gives lyophilisation of the appropriate fractions 2.5 g of the impure title compound in the form of a syrup. It is repurified by HPLC (micro-bondapak C-18) to give 0.90 g (7%) of the title compound.

1. R. (film) S> 2520 (SH), 1640 and 1625 (pyridinium), 20 1585, 1490, 1200 cm "1 (sulfonate), NMR H (DMSO-dg + D £ 0) c): 2, 36 (3H, s, CH3S03 "), 4.62 (2H, m, CH2N +), 7.74 (2H, m,

Hm of pyridinium), 8.24 (1H, m, Hp of pyridinium), U.V. (H20) Xmax: 272 (6 4080) m ^.

25 B. Paranitrobenzyl diphenylphosphate 3- Γ 2- (1- (2,6-dimethylpyridinium)) ethylthio1 -6oç - Çl- (R) -hydroxyethyl ~ 3 -7-oxo-1-azabicyclo (3.2.0) hept- 2 ene-2-carboxylate

OH Ό NEt (iPr) p OH

30 0 ά J ^

Vry0 2) cll! (QPh) *, COOPNB 3) HS— ^ Ms0. \ oopNB (Ph0) # 0 35 2) NEt (iPr) 2 104 To a cooled solution (0 ° C) of p-nitrobenzyl 6 <X- [1- (R) -hydroxyethyl3 -3,7-dioxo-1- azabicyclo (3.2.0) -heptane-2-carboxylate (0.658 g, 1.89 mmol) in aceto-nitrile (6 ml) maintained under a nitrogen atmosphere, diisopropylethylamine (0.394 ml, 2.26) is added mmol) and diphenyl chlorophosphate (0.468 ml, 2.26 mmol).

The reaction mixture is stirred for 30 minutes and treated with a solution of 1- (2-mercaptoethyl) -2,6-dimethylpyridinium methanesulfonate (0.720 g, 2.73 mmol) - in acetonitrile (3 ml) followed with diisopropylethyl amine (0.394 ml, 2.26 mmol). The reaction mixture is stirred at 0 ° C for 2 hours, eluted with cold water (0 ° C) (27 ml) and poured onto the top of a column (2.5 x 9.0 cm) of micro- bondapak C-18. Elution with mixtures of acetonitrile-water and freeze-drying of the appropriate fractions gives 0.92 g (65%) of the title compound, IR (KBr) Ό max: 3700-3000 (OH), 1765 (C = YS-lactam 0), 1690 -C = PNB ester), 1620 (pyridinium), 1590 (phenyl), 1517 (NO2), 1330 (NO2), 880 cm-1 (NO2) 1H NMR (DMSO , d6) Γ: 20 1.15 (3H, d, J = 6.2 Hz, 'CH3CH0H), 2.7-3.7 (11H, CH2S, 2-CH3 on pyridinium, H-4, H-6 ), 3.7-4.4 (2H, CH3CH0H, H-5), 4.7 (2H, m, CH2N +), 5.14 (1H, d, J = 4.5 Hz, OH), 5, 37 (center of ABq, J = 13.2 Hz, C ^ of PNB), 6.7-7.5 (10H, m, phenyl), 7.5-8.7 (7H, pyridinium, H's of PNB), UV (H20) X max: 25 274 (£ 14150), 319 (69445) myu .. ^ C. 3-L2- (l- (2,6-dimethylpyridinium) ethylthio] -6o (- C1- (R) - hydroxyethyl] -7-oxo-1-azabicyclo (3.2,0) -hept-2-ene-2-carboxylate

30 OH OH

X \, —V ^ \ Λα * „Pd / C> X 77λ \. rr> ~ # / $ buffer o ^ "/ COOPNB (PhO) 2P0 X300“

- O O

105 To a solution of p-nitrobenzyl diphenylphosphate 3-1 2- (1- (2,6-dimethylpyridinium)) - ethylthio] -6c <- (1 - (R) -hydroxy thyl] J -7-oxo-1 -azabicyclo (3.2.0) hept- 2-ene-2-carboxylate (0.80 g, 1.07 mmol) in wet tetrahydrofuran (42 ml) is added ether (42 ml), buffer monobasic potassium phosphate-sodium hydroxide (0.15 M, pH 7.22, 21 ml) and 10% palladium on charcoal (0.80 g) The resulting mixture is hydrogenated for 1 hour under a pressure of -jO 275.792 kPa at 23 ° C. and filtered through a pad of Celite, the two layers are separated and the organic layer is extracted with the pad (3 times 8 ml);

The aqueous phases are combined, washed with ether (50 ml), pumped to remove traces of organic solvent and poured onto the top of a column (3.0 x 10 cm) of micro-bondapak C-18. Elution from the column with a mixture of 5% acetonitrile-95% water and lyophilization of the appropriate fractions gives the title compound 0.246 g (63%) as a yellowish powder, 20 IR (KBr ) max: 3700-2800 (OH), 1750 (C = 0 de / 3 -lactam), 1620 (pyridinium), 1595 cm-1 (carboxylate), NMR (D20) S: 1,23 (3H, d, J = 6.4 Hz, CH3CH0H), 2.5-3.5 (11H, H-4, H-6, CH2S, 2CH3 on pyridinium), 3.8-4.4 (2H, CH3CH0H, H-5) , 4.5-4.9 (CH2N +, H0D), 7.64 and 7.74 (2H, part of the A2B system, pyridinium Hm), 8.07, 8.16, 8.18 and 8, 27 (1H, B part of the A2B system, pyridinium Hp), UV (H20) ^ max: 277 (* 9733), 300 (£ 8271) + 50.7 ° (C 0.48, H20),

Anal. wedge, for C ^ H ^ N ^ S 1.5 H 2 O: C 55.51, H 6.47, 30 N 7.19; found: C 55.14, H 6.23, N 6.46.

106 EXAMPLE 11

Preparation of (5R, 6S) -3 - (_ 2- (2-methylthio-3-methylimidazolio) ethylthio] [-6- '1- (R) -hydroxy-ethyl] 7-oxo-1-azat) icyclo ^ 3 "2" 03 hept-2-ene-2-carboxylate 5 OH * e N-Me CK \ CO® 10 2 A. 2-methylthio-3-methyl-1- (2-mercaptoethyl) imidazolium SMe 15 I \ trifluoromethanesulfonate \ '+ CF3S03H + _ ^ HSCH2CH2 n' N-Me

Nt ^ Sto W

Me CFjSOj® 20 Trifluoromethanesulfonic acid (1.38 ml, 0.015 mole) is added dropwise to 2-methylthio-1--1 methylimidazole (4.0 g, 0.03 mole) at 0 ° C under nitrogen ..

Ethylene sulfide (0.9 ml, 0.015 mole) is then added and the mixture is heated at 55 ° C under nitrogen for 24 hours. The reaction mixture is triturated with ether (3 times) and the residue is taken up in acetone, filtered and evaporated. This gives the product (4.2 g, $ 82) as a semi-crystalline solid which is used as it is without further purification. I.R. (film) \>: 2550 (w, sh) cm ”" ';

"4 IHcQC

30 N NMR (dg-acetone) 5: 7.97 (s, 2H), 4.66 (t, J = 7 Hz, 2H, methylene), 4.17 (s, 3H, N-Me), 3, 20 (d of t, J = 7 Hz, J '= 9 Hz, 2H, methylene), 2.72 (s, 3H, S-Me), 2.20 (t, J = 9 Hz, 1H, -SH ).

107 Λ

Prepared in accordance with A. Wohl and W. Marckwald, Chem,

Ber. 22, 1353 (1889).

B. P-nitrohenzyl diphenylphosphate (5R, 6S) -3- [2- (2-methylthio-5-methylimidazolio) ethyl thio] -5 6- 1- (R) -hydroxyethyl-7-oxo-1-azabicyclo [ 3.2.oj- hept-2-ene-2-carboxylate OH OH SMe

s 10 Vv_► VrVs ^ V

CO ^ PNB C02pnb (00) 2 ^ ° 15 To a solution of p-nitrohenzyle (5R, 6S) -6- (j- (R) -hydroxyéthylj -3,7-dioxo-1-azabicyclo [3.2.0 "! heptane-2-carboxylate (1.40 g, 4.0 mmol) in 50 ml of anhydrous acetonitrile, at 0 ° C. under nitrogen, diisopropylethylamine is added dropwise (0.76 ml, 4.4 mmol ) followed by diphenyl chlorophosphate (0.91 ml, 4.1 mmol).

After stirring the reaction mixture at room temperature for 1 hour, diisopropylethylamine (0.76 ml, 4.4 mmol) is added and then a solution of 2-methylthio-3-methyl-1 trifluoromethanesulfonate is added dropwise - (2-mercaptoethyl) imidazolium (2.0 g, 5.9 mmol) in 5 ml of acetonitrile. The reaction mixture is left at room temperature for 1.5 hours and then concentrated in vacuo to give a gum.

This gum is taken in H2O and applied to a C ^ g reverse phase column. Elution with water, then 20% acetonitrile-H2O and finally 30% acetonitrile-H2O, followed by lyophilization of the appropriate fractions results in the product (0.90 g, 30%) in the form a solid; 35 light yellow. I.R. (KBr) S): 3380 (br, OH), 1770

IlclX

108 (/ ¾-lactarne CO) cm-1; 1H NMR (dg-acetone) <$: 8.35 (br, s, 1H), 8.24, 6.78 (AB q, J = 8.8 Hz, 4H, aromatic), 7.89 (br, s, 1H), 7.25-6.91 (m, 10H, diphenylphosphate), 5.50, 5.25 (AB q, J = 12 Hz, 2H, benzyl), 4.75-4.27 (m , 3H), 4.03 5 (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-C 2- (2-methylthio-3-methylimidazolio) - ethylthio ^ -6- | "1 - (R) -hydroxyethyl] -7-oxo-10 azabicyclo [3.2,0J hept -2-ene-2-carboxylate OH * ef

”^„ W

\ o2PNB (0O) 2PO C02® 2o To a solution of p-nitro-benzyl diphenylphosphate (5R, 6S) -3- [2- (2-methylthio-3-methylimidazolio) -ethyl thio! [-6- [_1 - (R) -hydroxy thyl J -7-oxo-1-azabicyclo- [3.2.0 1 hept-2-ene-2-carboxylate (1.20 g, 1.56 mmol) in a mixture of 70 ml of THF , 70 ml of ether and 31 ml of phosphate buffer (0.05 M, pH 7.4) 1.2 g of 10% palladium on carbon are added. This mixture is hydrogenated (Parr) at a pressure of 241.318 kPa for 55 minutes. The reaction mixture is then filtered through a pad of Celite and the filter cake is washed with HgO and ether. The aqueous phase is separated, cooled to 0 ° C and the pH is adjusted to 7.0 with saturated aqueous NaHCO 4. After removal of the residual organic solvents in vacuo, the aqueous solution is applied to a reverse phase column at C ^ g. Elution with HgO and then 8% aceto-nitrile-O and subsequently lyophilization of the appropriate fractions 109 gives 0.25 g of a solid. This material is repurified by reverse phase HPLC to give the product (0.114 g, 19%) in the form of a practically white solid.

IR (KBr) ^ max: 3420 (OH), 1750 (ß-lactam CO), 1590 5 (-¾0) cm "1; 1 H NMR (D20) Γ: 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 (H20) Xmax: 297 (6 7572), 262 U 6259), 222 ( t 7955) nm.

V, 10 EXAMPLE 12

Preparation of (5R, 6s) -5-L2- (3-aminopyridinio) -ethyl thio] -6- Q1- (R) -hydroxyethyll -7-oxo-1-azabicyclo [3 "2.0] hept-2-ene- 2-carboxylate

OH NH

/ - N ._ // N Θ co® 20 A. 3-amino-1- (2-mercaptoethyl) pyridinium chloride H £ N .. H2N _ / NH2

- 25 Y "J +) ¾ + HSCH2CH

'' N .HCl \ \ ^ = y

Cl®

3-Aminopyridine (1.50 g, 0.016 mole) is taken in 15 ml of 1 N methanolic HCl and the resulting solution is evaporated to give the hydrogen chloride product as an oil. To this oil was added 3-aminopyridine (1.32 g, 0.015 mole) and ethylene sulfide (0.89 ml, 0.015 mole) and the resulting mixture was heated (oil bath) to 60 -65 ° C under nitrogen for 2 hours.

110

Another equivalent of ethylene sulfide (0.89 ml, 0.015 mole) is added and heating is continued at 55-65 ° C for 65 hours. The reaction mixture is washed with CH2Cl2 and then taken in H2O (25 ml). The aqueous solution is applied to a C ^ g reverse phase column which is eluted with ΕΡ, Ο. Evaporation of the appropriate fractions gives the product (1.26 g, 44%) as a colorless viscous oil. I.R. (film) 0: 3180 (NH „) cm; H NMR (dg-DMSO) o: 8.19-7.59 (m, 4H, aromatic), 4.59 'J 10 (t, J = 2 HZ, 2H, N-CH2), 3.5 (hr s, 2H, -NH2), 3.20-2.77 (m, 3H).

s> B. p-nitrobenzyl diphenylphosphate (5R, 6S) -3- [2- (3-aminopyridinio) ethyl thio]] -6- £ 1- (R) -hydroxyethylj -7-ox: o-1-azabicyclo (3.2.0) hept-2-15 ene-2-carboxylate

NH

OH OH / 2! 0 το - * -

C02PNB C02pnb (0O) 2PO

25 To a solution of p-nitrobenzyl (5R, 6S) -6- (1- (r) - hydroxyethyl) -3,7-dioxo-1-azabicyclo (3.2.0) heptane-2-carboxylate (0.696 g, 2 , 0 mmol) in 10 ml of anhydrous acetonitrile, at 0 ° C. under nitrogen, diisopropylethylamine (0.382 ml, 2.2 mmol) is added dropwise followed by diphenyl chlorophosphate (0.457 ml, 2.2 mmoles). After stirring at 0 ° C for 30 minutes, a solution of 3-amino-1- (2-mercaptoethyl) pyridinium chloride (0.475 g, 2.5 mmol) in 1 ml of anhydrous DMF is added, followed by additional diisopropylethylamine (0.435 ml,

35 2.5 mmol). Maintaining the reaction mixture at 0 ° C.

111 for 1.5 hours and then concentrated in vacuo. The resulting gum is taken in acetonitrile-H ^ O (1: 1) and applied to a reverse phase column in c1g. Elution with water, followed by 20% acetonitrile-H2O and subsequent lyophilization of the appropriate fractions results in the product (0.730 g, 50%) as a beige solid. LR. (KBr) i) max: 3330 (br, OH), 3180 (br, NH2), 1770 (-lactam CO), 1690 (-C02PNB) cm-1; 1H NMR (dg-DMSO) 5: 8.29-7.63 (m, 8 aromatic), 7.2-6.7 (m, 10H, diphenylphosphate), 5.47-5.18 (AB d, J = 14 Hz, 2H, benzy-10 lique), 4.73-4.45 (m, 3H), 4.2-3.8 (m, 1H), 3.6-2.6 (m, SH) , 1.15 (d, J = 6.2 Hz, 3H, CHMe).

C. (5R, 6S) -3- £ 2- (3-aminopyridinio) ethyl thio] -6- \ 1- (R) -hydroxyethylJ -7-oxo-1-azabicyclo (3.2.0) -15 hept-2 -ene-2-carboxylate OH HH, ° TH / H2 /) —- J-14 ^ ^ \ fi CO®

C02PNB (0O) 2PO

25 To a mixture of p-nitro-benzyl diphenylphosphate (5R, 6S) -3- [2- (3-aminopyridinio) ethyl thio ^ -6- [1- (R) -hydroxyethyl] -7-oxo-1- azabi cyclo (3.2.0) hept-2-ene-2-carboxylate (0.730 g, 1.0 mmol) and 10% palladium on carbon (0.7 g) in 25 ml of phosphate buffer 30 (0.05 M, pH 7.4) 8 ml of THF and 20 ml of ether are added.

This mixture is then hydrogenated (Parr) at a pressure of 275.792 kPa for 1 hour. The resulting mixture is filtered through a pad of Celite and the filter cake is washed with H2O and ether. The aqueous phase is separated, 112 washed with ether (2 times) and then the residual volatiles are removed in vacuo. The aqueous solution is immediately applied to a C ^ g reverse phase column which is eluted with H ^ O. Lyophilization of the appropriate fractions resulted in 0.45 g of a practically white solid. This material is repurified by reverse phase HPLC to give the desired product (0.123 g, 35%) in the form of an ivory-colored solid.

IR (KBr) Ό max: 3340 (br), 1750 (br, / 3-lactam CO), 10 1580 (br, -002θ) cm "1; 1 H NMR (DgO) £: 8.07-7.59 ( m, 4H, aromatic), 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 of t, J = 9.2 Hz, J' = 2.6 Hz, 1H, H-5), 3.38 (t, J = 5.8 Hz, 2H, S -CH2), 3.24 (d of d, J = 6.0 Hz, J '= 2.6 Hz, 1H, H-6), 3.17-2.57 Cm, 2H, H-4), 1.21 (d, J = 6.3 Hz, 3H, CHWe); U.V. (HgO) 299 (8,794), 256 (t 8822) nm; t ^ (pH 7.4, 36.8 ° C) 18.5 h.

EXAMPLE 13 PREPARATION OF

20 ^ OH

0 ^ © 25 C0 ° (5R, 6s) 3-1 1- (S) -methyl-2- (1-pyridinium) ethylthiol -6- 11- (R) -hydroxyethyl] [-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-30 2-carboxylate and c? 113

OH

T. - / 7Λ 'vile /'. ln

^ 'Q

coo ^ (5R, 6S) 3- [1 (R) -methyl-2- (1-pyridinium) ethylthio] - 10 6- [l- (R) -hydroxyethyl ^] - 7-oxo-1-azabicyclo ( 3.2.0) hept- 2-ene-2-carboxylate A. dl-1- methanesulfonate (2-mercapto-2-methylethyl) - pyridinium dl-1- methanesulfonate (2-mercapto-1-methylethyl) - pyridinlum

Ms0_ 20 A + ι / ζ)) + Ms0H ___ ^ C +) \ MsO “hs · '^^ \ _n / 25

Methanesulfonic acid (1.95 ml, 0.030 mole) is added slowly to cold pyridine (7.83 g, 0.097 mole) and the resulting mixture is stirred at 40 ° C for 15 minutes, treated with sulfide dl-propylene (2.59 ml, 0.033 mole) and stirred at 60 ° C under nitrogen for 90 hours. The pyridine is removed under vacuum; the residue is mixed with water and purified by 3 · chromatography (hplc, Prep. Bondapak C-18). The appropriate fractions are combined and lyophilized to give dl-1- (2-mercapto-2-methylethyl) pyridinium methanesulfonate 1.14 g (15%) as a colorless syrup; 114 IR (film) γ>: 2520 (SH), 1640 (pyridinium), 1180 (s, CH3S03 ~), 1040 (CH3S03 ~) cm-1, 1H NMR (DMSO dg) c: 1.35 (d, J = 6.8 Hz, 3H, CH3CHS), 2.30 (s, 3H, CH3S03 “), 2.90 (d, J = 8.5 Hz, 1H, SH), 3.2-3.7 (m , CHSH), 4.52 (dd, Jgem = 12.9 Hz, 5 J = 8.4 Hz, CHCH0N +), 4.87 (dd, J = 12.9 Hz, J = 6.0 Hz,, - gern 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 (HgO) Xmax: 208 (£ 5267), 259 (t-3338),

Anal, hold, for CÿH | 3N03S2.2H20: C 37.88, H 6.71, 10 N 4.91, S 22.47; found: C 37.49, H 6.85, N 4.86, S 22.09 and dl-1- (2-mercapto-1-methyl-ethyl) pyridinium methanesulfonate 0.82 g (11%) in the form a colorless syrup; IR (film) \): 2500 (SH), 1628 (pyridinium), 1180 (sulfonate), 1035 (sulfonate) cm NMR H (DMSO dg) ù: 15 1.69 (d, J = 6.8 Hz, 3H , CH3CHN +), 2.31 (s, 3H, CH ^ O ^), 3.0- 3.3 (m, 2H, CH2S), 4.2-5.2 (m, 1H, CHN +), 8.0- 8.4 (m, 2H,

Hm of pyridinium), 8.5-8.8 (m, 1H, Hp of pyridinium), 9.0- 9.2 (m, 2H, Ho of pyridinium), U.V. (Ho0) λ: 209

f <L In 8.X

(£ 4987), 258 (* 3838). Anal, wedge, for 0 ^^ 0 ^ .1.5 ^ 0: 20 C 39.11, H 6.56, N 5.07; found: C 39.13, H 5.92, H 5.20.

B · (5R, 6S) paranitrobenzyl diphenylphosphate 3- [1- (R, S) methyl-2- (1-pyridinium) ethylthio] -6-Ï.1 - (R) -Uydroxyéthyl ^ -7-oxo-1 -azaMcyclo (3.2, 0) -25 hept-2-ene-2-cart> oxylate n „D NEt (iPr)„

® o 2 OH

0 ^ N 3) hs — C-î (O) Ms0 "COOPNB COOPNB,. · T _ 4) NEt (iPr) 2 (ph0) 2P0 .7 115 To a cooled solution (0 ° C) of (5R, 6S ) para-nitrobenzyl 6- 1- (R) -hydroxyethyl J -3,7-dioxo-1-aza-bicyclo (3-2.0) heptane-2-carboxylate (0.523 g, 1.5 mmol) in acetonitrile ( 6 ml) maintained under a nitrogen atmosphere, diisopropylethylamine (0.314 ml, 1.8 mmol) is added followed by diphenyl chlorophosphate (0.373 El, 1.8 mmol), the reaction mixture is stirred for 30 minutes and treated with a solution of dl-1- (2-mercapto-2-methylethyl) -10 pyridinium methanesulfonate (0.539 g "2.16 mmol) in acetonitrile (2 ml) and diisopropylethylamine (0.314 ml, 1, Mm The reaction mixture is stirred at 0 ° C for 1 hour, diluted with cold water (0 ° C) (24 ml) and chromatographed on a prep column bondapak C-18 (2 , 5 x 8.5 cm) with 15 as elution solvents 25-50% acetonitrile in water to give, after lyophilization, 1.07 g (97%) of the title compound in the form of a yellowish powder.

IR (KBr) 0max: 3700-3100 (OH), 1770 (C = 0 of -lactam), 1695 (C = 0 of the PNB ester), 1630 (pyridinium), 1590 (phenyl), 20 1518 (N02) , 1348 (N02) 885 (N02) cm "1, 1 H NMR (dMSO dg) S: 1.14 (d, J = 6.1 Hz, 3H, CH ^ QHO) 1.33 (d, J = 6, 3 Hz, 3H, CEUCHS), 4.6-5.0 (m, CH2N), 5.14 (d, J = 5.2 Hz, 1H, OH), 5.37 (center of ABq, J = 12 , 4 Hz, 2H, 0Η £ of GNP), 6.6-7.5 (m, 10H, phenyl phosphate), 7.69 (d, J = 8.7 Hz, 2H, Ho of PNB),

25 8.0-8.4 (m, 4H, Hm of GNP, Hm of pyridinium), 8 4-8 8 (m 1H

Hp de pyridlni ”), 9> 08 (d, .5> 6 Hz, 2H> hÎ de pyïdini ^) U.V. (H20) ^ max: 263 (fc13325) 308 (Ç8915). Anal, hold, for C36H36N3010SP.H20: C 57.52, H 5.10, H 5.10, N 5.59, S 4.27; found: C 57.76, H 4.96, N 5.35, s 4.35, 116 C. (5R, 6S) 3- L1- (R and S) -methyl-2- (l-pyriainium) - ethylthioj -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3 * 2.0) hept-2-ene-2-carboxylate

5 OH OH

X,, - \ 10% Pd / C, XIJ ('2 · a _ & ___ * p pL / s ™? 4gsr 0 ^ ~ N ^ (^ 0 \ (PhO) 9PO ~ C00 “10 COOPNB ^ To a solution of diphenylphosphate of (5R, 6S) -paranitrobenzyl 3- 1- (R, S) -methyl-2- (1-pyridinium) ethylthiq} 15 -6-Γ1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo ( 3.2.0) hept-2- ene-2-carboxylate (0.60 g, 0.82 mmol) in wet tetrahydrofuran (33 ml) ether (33 ml), phosphate buffer are added monobasic potassium-sodium hydroxide (17 ml, 0.15 N, pH 7.22) and 10% palladium on charcoal (0.60 g) The resulting mixture is hydrogenated for 1 hour at a pressure of 275,792 kPa at 23 ° C.

The two layers are separated and the organic layer is extracted with water (3 times 7 ml). The aqueous layers are combined, filtered through a pad of Celite, washed with ether (3 times 20 ml) and chromatographed on a prep column. bondapak C-18 (2.5 x 9.5 cm) with water as the elution solvent to give 0.18 g (63%) of mixture of diastereoisomers. The two diastereoisomers are separated by hplc (prep. Bondapak C-18) with 3Q water as elution solvent: isomer with the lowest retention time, 0.068 g (23%) compound "B", IR ( KBr) O: 1770 (C = 0 of 7 ^ -lactarne), 1633 (pyridinium), ~ max λ 1593 (carboxylate) cm, NMR H (D £ °): 1.20 (d, J = 6.3 Hz , 3H, CH3CH0), 1.42 (d, J = 6.9 Hz, 3H, CH ^ CHS), 2.3-3.2 (m, 3H, 117 H-4, H-6), 3, 5-3.9 (m, 1H, SCH), 3.9-4.2 (m, 2H, H-5, CH ^ CHO), 4.3- 5.1 (m, CH2N +), 7.8-8 , 2 (m, 2H, Hm of pyridinium), 8.4- 8.7 (m, 1H, Hm of pyridinium), 8.7-9.0 (m, 2H, Ho of pyridinium), UV (HgO) ffiax: 260 ¢ .6727), 300 ¢ 8245), ^ '^ p®)> 1 - 12.6 h (measured at a concentration of 10 M in phosphate buffer pH 7.4, at 36.8 ° C); isomer with the longest retention time, 0.081 g (28%), compound "A", IR (KBr): 1755 (C = 0 of; f) \ max a-lactam), 1630 (pyridinium) , 1590 (carboxylate) cm ”, 10 1 H NMR (D20) S: 1.18 (d, J = 6.3 Hz, 3H, CHÿiHO), 1.40 (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, CH ^ CHO, H-5), 4.2-5.1 (m, CH2N +), 7.7-8.1 (m, 2H, Pyridinium Hm), 8.3-8.65 (m, 1H, Hp pyridinium), 8.65-8.9 (m, 2H, pyridinium Ho 15), UV (H? 0) λ: 259 (6,5694), 296 ¢ 6,936), [pOp + 96.9 ° (c 0.56, H2O), t ^ = 15.6 h (measured at a concentration of 10 “^ M in phosphate buffer pH 7.4 at 36.8 ° C).

EXAMPLE 14

20 PREPARATION OF

OH

25 '25 yy — N__y tr 0 ^ \ 5 C00 “. NOT-.

(V) (5R, 6S) (S) - (1 -pyridinium) 1 -1- (S) -cyclohexylthio "3 -6- ^ 1- (R) -hydroxyethyl-7-oxo-1-azabicyclo ( 3.2.0) hept-2-ene-2-carboxylate r 30 118

and OH

'V-j-'v I, - SMI.l 1 • 5

O -> A

/ Nv (©.! -3 10 (5R, 6S) 3- [2- [(R) - (l-pyridinium) 3 1- (R) -cyclo-> hexylthiol -6- {_ 1- (R) -hydroxyethylj -7-oxo-1- azabicyclo (3.2.0) hept-2-ene-2-carboxylate 15 A. dl-1- (2-mercapto-1-cyclohexyl) methanesulfonate - pyridinium

0 SH

; MsO "_ 25

Methanesulfonic acid (0.65 ml, 0.01 mole) is added dropwise to the pyridine (2.42 ml, 0.03 mole) under cooling. The mixture is stirred under a nitrogen atmosphere for 10 minutes, treated with dl-30 cyclohexene sulfide £ 1.377 g (85% pure), 0.0102 mole ^ and stirred at 72 ° C for 25 hours. The excess pyridine is removed in vacuo and the traces are codistilled with water. The residue is mixed with water and chromatographed through a prep column. bondapak C-18 (5 x 13 cm) with -2 35 eluting solvent 0-2% acetonitrile in water, giving after 119 freeze-drying a colorless syrup, 1.57 g (53%), IR (film ) v max: 2500 (SH), 1625 (pyridinium), 1190 (S0 ^ ~), 1 H NMR (DMSO dg) l): 1.2-2.5 (m, 8H, cyclohexyl H) 2.32 (s , 3H, CH3S03 “), 2.82 (d, J = 9.8 Hz, SH), 3.0-3.5 (m, 1H, CHSH), 5 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 de pyridinium), UV (¾0) Λ max: 214 (1 5365), 258 (^ 3500),

Anal, cal. for 2 ^ 19 ^ 3¾ * ^ 2 ^: ^ ^ 6.88, H 6.88, N 4.56; found: C 46.61, H 6.46, N 4.65.

10 B. (5R, 6s) paranitrobenzyl diphenylphosphate 3- 5 \ 2- 2- L (R or S) - (l-pyridinium) 3 -1- (R or S) -cyclohexylthiol -6 - {_ 1 - ( R) -hydroxyethyll -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carhoxylate 15

OH 1) NEt (iPr) 2 0H

î 8 T

’ΝΓΤ ^ \ - Z) cip (0ph) 2" 9- ^ 20 3) Hs-p] L- oXLf-3'g_

COOPNB / —COOPNB

@ 10.1 4) NEt (iPr) 2 25 To a cooled solution (0 ° C) of (5R, 6s) para-nitrobenzyl 6-Il 1- (R) -hydroxyethyll -3,7-dioxo-1-azabicyclo- ( 3.2.0) heptane-2-carboxylate (1.37 g, 3.93 mmol) in acetonitrile (15 ml) maintained under a nitrogen atmosphere 30 diisopropylethylamine (0.822 ml, 4.7 mmol) is added and diphenyl chlorophosphate (0.979 ml, 4.7 mmol).

The resulting solution is stirred for 30 minutes and treated with a solution of dl-1- (2-mercapto-1-cyclohexyl) pyridinium methanesulfonate (1.64 g, 5.66 mmol) in acetonitrile (4.7 ml) followed by diisopropylethylamine 120 (0.822 ml, 4.7 mmol). The reaction mixture is stirred at 0 ° C for 1 hour, diluted with cooled water (0 ° C) (75 ml) and chromatographed on a prep column. bondapak C-18 with 25-50% acetonitrile in water as elution solvent, giving, after lyophilization, appropriate fractions 1.9 g (53%) of the title compound, I.R. (KBr) Ό:

ÎÎIcLX

3700-3000 (0H), 1770 (C = 0 of / 3-lactam), 1700 (C = 0 of the PNB ester), 1628 (pyridinium), 1590 (phenyl), 1515 (N02), 1345 (N02) , 880 (N02) cm-1, 1H NMR (D20) S: 1.13 (d, 10 J = 6.1 Hz, 3H, CH ^ CHO), 1.2-2.5 (m, 8H, H cyclohexyl), 2.7-3.5 (m, 4H, H-4, H-6, CHS), 3.5-4.4 (m, 2H, CH3CH0, H-5), 4.4- 5.0 (m, 1H, CHN +), 5.30 (center of ABq, J = 12.8 Hz, CH2 of PNB), 6.7-7.4 (m, 10H, phenyl), 7.65 ( d, J = 8.6 Hz, 2H,

PNB Ho), 7.9-8.4 (m, 4H, PNB Hm, pyridinium Hm), 8.4-8.8 (m, 1H, pyridinium Hp), 9.0-9 , 4 (m, 2H, Ho of pyridinium), UV (Η £ 0) Xmax: 263 (£ 9038), 309 (£ 6394).

Anal, wedge, for C ^ H ^ qN ^ O ^ qSP. ^ O: C 59.16, H 5.35, N 5.31; found: C 58.95, H 5.15, N 5.57.

20 C. (5R, 6S) 3-L 2- I (R or S) - (1-pyridinium) 3 -1- (R or S) - cyclohexylthio]] -6- £ 1- (R) -hydroxyethyl] -7-oxo-l-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

or OH OH

J l

f j 10% Pd / C, H2 x 4 — I

’—I // S THF, ether, buffer ^ -Nv / -3 0 ^ \! 0 \ Nv COOPNB ^ N. 0 C00 ~ i7? Vl

30 (Ö1 (Ph0) 2 ^ 'l ^ J

-Μ * 121 To a solution of (5R, 6S) para-nitrobenzyl diphenylphosphate 3-2- {_ (R or S) - (1-pyridinium) J -1- (R or S) -cyclohexylthioj -6- [ 1- (R) -hydroxyethyl \ -7-oxo-1-azabicyclo ~ (3.2.0) hept-2-ene-2-carboxylate (1.85 g, 2.34 mmol) in 5 of wet 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 ml) are added g) The resulting mixture is hydrogenated at 23 ° C under a pressure of 275,792 kPa (40 psi) for 1.5 hours. The organic layer is separated and washed with water (3 times 20 ml). The aqueous solutions are filtered through a pad of Celite, with ether (2 times 60 ml), pumped to remove the traces of organic solvents and chromatographed on a prep column.

15 bondapak C-18 (4.5 x 9 cm) with the elution solvent 0-5% acetonitrile in water, after lyophilization giving 0.705 g (76%) of a mixture of diastereoisomers. The separation of the diastereoisomers is carried out by hplc (prep. Bondapak C-18) with as elution solvent 4% 20 of acetonitrile in water; the diastereoisomer with the lowest retention time, compound "A", (0.29 g, 31%), IR (KBr) -O "max: 1750 (C = 0 of / ^ - lactam), 1620 (sh , pyridinium), 1685 (carboxylate) cm- ^, NMR (D20): „1.21 (d, J = 6.3 Hz, 3H, CH ^ CHO), 1.4-2.5 (m, 8H, Cyclohexyl H), 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, CH ^ HO), 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) Xmax: 260 (6.7123), 300 (68685), + 6.2 ° (c 30 0.63, H90), t ^ = 16.6 h (measured at a concentration of -4 ^ 2 10 M in phosphate buffer pH 7 , 4-36.8 ° C);

Anal, hold, for C20H24N2 ° 4S * 2H2 °: 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; diastereoisomer with the longest retention time, compound "B", (0.35 g, 38%), IR (KBr) v> max: 1750 (C = 0 of 122 / i-lactam), 1662 (sh, pyridinium), 1588 (carboxylaxis) cm 1, 1 H NMR (D20) £: 1.19 (d, J = 6.4 Hz, 3H, CH ^ CHO), 1.3-2.5 (m, 8H, Cyclohexyl H), 2.5-3.1 (m, 2H, H-4), 3.1-3.3 (m, 1H, H-6), 3.3-3.8 (m, 2H , H-5) CHS), 4.1 (center of m, 1H, 5 CH3CHO), 4.25-4.7 (m, 1H, CHN +), 7.8-8.1 (m, 2H, Hm pyridinium), 8.3-8.7 (m, 1H, pyridinium Hp), 8.75-9.0 (m, 2H, pyridinium Ho), UV (H90) X: 259 (f5992), 296 (i 7646), | ÇXjyJ +65.3 “(ç 0.43, HgO), ti = 20.2 h (measured at a concentration of 10-½ in 10 phosphate buffer pH 7.4 at 36.8 ° C), EXAMPLE 15 A. (5R) allyl diphenylphosphate 3-Ç (2-pyridinioethyl) thioQ - (6S) - f (lR) -hydroxyethyl3-7-oxo-1-15 azabicyclo 13.2.03 hept-2 -ene-2-carboxylate

OH | H

1) C1P0 (0s <) Q / n \ hs v_y co ^> © 0po (cV) 2 25 To a solution of (5R) allyl 3,7-dioxo- (6S) - £ (lR) -hydroxyethyl -1- azabicyclo 13.2.03 heptane- (2R) -carboxylste (473 mg, 1.87 mmol) in CH ^ CN (6 ml) is added at about -10 ° C under nitrogen atmosphere of diisopropylethylamine 30 (0.42 ml , 2.4 mmol) followed by diphenyl chlorophosphate (0.50 ml, 2.4 mmol). The mixture is stirred at 0 ° C for 30 minutes, and then cooled to -15 ° C. To this is added an oily suspension of M (2-mercapto-ethyl) pyridinium chloride (527 mg, 3.00 mmol) in CH ^ CN (1 ml) containing 5 drops of DMF, followed by 123 diisopropylethylamine (0.42 ml, 2.4 rnmoles). The mixture is stirred at -15 ° C for 30 minutes and then diluted with H2O (20ml). This mixture is directly purified on a reverse phase silica gel (PrePAK, 12 g, trade name of Waters Associates), eluting with water (200 ml), 10 ° / o of CH CN / HgO (100 ml ), 20% CH3CN / H20 (100 ml), 30% CH-CN / H 0 (100 ml) and then 40% CH, CN / H ^ O (100 ml).

3 2 ^ 3 2

The appropriate fractions are collected, the organic solvent is removed by a vacuum pump and lyophilized to obtain 786 mg (1.26 mmol, yield 67.3%) of the title compound as a brownish powder.

1H NMR (DMSO-dg, CFT-20) c: 1.16 (3H, d, J = 6 Hz, 1'-CH3), 2.6-3.7 (m), 3.75-4.3 (2H, m, 5-H and 1'-H), 4.65 (2H, m, -C02CH2-), 4.87 (2H, t, J = 6 Hz, -CH2-N +), 5-6 , 2 (3H, m, 15 olefinic protons), 6.6-7.4 (m, aromatic protons), 8.15 (2H, "t", J = 7 Hz, aromatic protons meta with respect to nitrogen ), 8.63 (1H, "t", J = 7 Hz, aromatic proton para with respect to nitrogen) and 9.07 ppm (2H, "d", J = 7 Hz, aromatic provons ortho with respect to nitrogen); I.R. (film) 0: 20 3400 (OH), 1770 (/ i> - lactam), 1690 (ester), 1625 (pyridinio).

. (5R) 5- L 2-pyridinioethyl) thio3 - (6g) - [(1R) ~ hydroxyethyl] -7-oxo-1-azabicyclo [3-2.0J hept- 2-ene-2-carboxylate 25

OH

y oh

"Vr \ STh Μ (ρ44 —— \ ® / ^ \ CO ^ 'V' ®0Ρ0 (0 $, ^ N ^ VC02K ^ Yqq i #! Ii 124 A solution of (5R) allyl 3-L diphenylphosphate ( 2-pyridinioethyl) thio] - (6S) - [(IR) -hydroxyethyl -7-oxo-1-azabicyclo [3.2.0J hept-2-ene-2-carboxylate (156 mg, 0.25 ffimole) in CH ^ CN (2 ml) is successively added 5 at approximately 22 ° C. a solution of potassium 2-ethylhexanoate in EtOAc (0.5 M, 0.6 ml; 0.3 mmol), triphenylphosphine (15 mg, 0.057 mmol) and tetrakis-triphenylphosphine palladium (15 mg, 0.013 mmol) The mixture is stirred at approximately 22 ° C. under a nitrogen atmosphere for 10 hours. After addition of anhydrous Et 2 O (7 ml), the filter is filtered. precipitated, washed with anhydrous Et ^ O (7 ml) and dried under vacuum to obtain 101 mg of brownish solid. This is purified by reverse phase column chromatography (PrePAK C ^ g, 12 g, from Waters Associates) eluting with H20.

The appropriate fractions (fractions 7-12, each of 20 ml) are collected and lyophilized to obtain 53 mg (0.16 mmol, yield 64%) of the title compound as a yellowish powder. This material is contaminated with potassium diphenylphosphate and potassium 2-ethylhexanoate: 1 H NMR (D20, CFT-20) <f: 0.80 (t, J = 6.4 Hz, Me of ethylhexanoate), 1.21 (3H, d, J = 6.3 Hz, 1'-Me), 2.93 (2H, dd, J = 9 Hz, Jgem = 4 Hz, 1-Hs), 3.28 (1H, dd,, = 6.2 Hz, Î6-5 = 2'5 Hz '6 "H)' 3'42 (2H> t» £ = 6 Hz '”CH2S)» 3.98 (1H, td, £ 5 -1 = 9 ^ z '% -6 = 2'5 Hz' 5 “H) '4'15 (1H' 9- '^ = 6'2 Hz' 25 4.80 (2H, t, J = 6, 0 Hz, -CH2N +), 7-7.5 (m, phenyl protons of diphenyl phosphate), 8.03 (2H, m, Hm of pyridinium), 8.56 (1H, m, Hp of pyridinium) and 8 , 81 ppm (2H, "d", J = 6.5 Hz, Ho of pyridinium).

; 125 ί EXAMPLE 16

Preparation of 3- [2- (N-methyl-thiomorpholinium) -ethylthio] -6c \ -L1 '- (R) -hydroxyethyll -7-oxo-1-azabicyclo l3.2.0] hept-2-ene-2-carboxylate 5

OH

T

^ Θ / \ J— S Me 1O 0 © C02 ^ 'la »A. Methanesulfonate N-methyl-N- (2-mercaptoethyl) - thiomorpholinium 15 _. Me / \ 1) Ms0H HS \ / \

MeN S --—- ► \ / \ jn S

^ - / 2) S. © \ -f 2o ^^ MsO ® A of precooled N-methylthiomorpholine (ice bath) (5.00 g, 42.7 mmol) methanesulfonic acid is added (1.47 ml, 20.5 mmol) and ethylene sulfide (1.30 ml, 21.4 mmol). The mixture is heated at 65 ° C for 24 hours and diluted with water (25 ml). The aqueous solution is washed with diethyl ether (3 times 25 ml), pumped in vacuo and poured onto a reverse phase column with silica gel; the title compound 30 is eluted with water. The appropriate fractions are combined and evaporated to produce the thiol as an oil (4.80 g, 86% yield); I.R. (film):

λ # 1 - Ulâ-X

2550 cm "1 (w, SH); NMR H (DMSO dg) 5: 3.25-2.95 (6H, m, CH2 #), 3.32 (3H, s, CH3 #), 3.20- 2.65 (7H, m, CH2S, SH) and 2.32 ppm (3H, s, CH ^ SO ^).

j j j 126 * 4f · J.M. Lehn and J. Wagner. Tetrahedron, 26, 4227 (1970).

B. para-nitrobenzyl 3- L 2- (N-methyl-thiomorpholinium-diphenylphosphate) ethylthio 3 - 6 oç - C 1 '- (R) -5-hydroxy-ethyl]) -7-oxo-1-azabicyclo L 3.2. 0} hept-2-ene-2-carboxylate

OH

OH 1) EtN (—C) 2 jf Y ° i g ^ C02PNB m ^ \ _ v co2pnb (^ ç (o ^) 2 b 4) EtN (- <) 2 15

A cold solution (ice bath) of para-nitrobenzyl 6 (\ - C1 '- (R) -hydroxyethyl] -3,7-dioxo-1-azabicyclo .3.2.0] heptane-2-carboxylate is treated (557 ml, 1.60 mmol) in CH- ^ CN (8 ml) dropwise with diisopropylethylamine (0.336 ml, 1.92 mmol) and diphenyl chlorophosphate (0.400 ml, 1.92 mmol) and the mixture is stirred for 30 minutes. The reaction mixture is treated with N-methyl 1-N- (2-merc apto ethyl 1) thio-morpholinium methanesulfonate (893 mg, 2.29 mmol) in CH 3 CN (4 ml) and diisopropylethylamine (0.336 ml, 1.92 mmol) and stirred for 30 minutes The solution is diluted with water (20 ml) and poured onto a reverse phase column of silica gel. The desired compound is eluted with a mixture of 50% aceto-nitrile-water. The appropriate fractions are combined, pumped under vacuum for 2 hours and lyophilized to produce the title compound (1.01 g, 85% yield): IR (nujol) y) max: 1760 (s, fi -lactarne C = 0) and 1510 cm-1 (s, N02); 1H NMR (DMSO-dg) &: 8.25 (2H, d, J = 8.8 Hz, 35 H-aromatic), 7.70 (2H, d, J = 8.8 Hz, H-aromatic), 127 7.33-6.84 (10 H, m, H-aromatic), 5.37 (2H, center of ABq, J = 14.2 Hz, CH2), 5.14 (1H, d, J = 4 , 5 Hz, OH), 4.35-3.80 (2H, m, H-1 'and H-5), 3.75-3.45 (6H, m, CH2N +), 3.31 (3H, s, CH3N +), 3.45-2.75 (9H, m, CH2S, H-6 and H-4) and 1.15ppni (3H, 5 d, J = 6.2 Hz, CH ^).

C. 3- Γ. 2- (N-methyl-thiomorpholinium) ethylthid] -6o (- 1 * - (R) - hydroxyethyl 1-7-oxo-1-azabicyclo L3.2.o3-hept-2-ene-2-carboxylate 10

OH

T 0H

Vt "\ Me H K .x M

15 Y] COpPNB Q JpP) 2% ^ (o ^) 2 CC ^ 0 20 A solution of para-nitrobenzyl 3- [_ 2- (N-methyl- thiomorpbolinium diphenylphosphate) ethylthio 3 -6 <3 - \ _1 '- (R) -hydroxyethyl ^ -7-oxo-1-azabicyclo ^ 3.2.θ] hept-2-ene-2-carboxylate (1.31 g, 1.76 irnnole) in 0.1 M phosphate buffer pH 7.4 (48.8 ml), tetrahydrofuran (20 ml) and diethyl ether (20 ml) is hydrogenated over 10% palladium / C (1.5 g) in a Parr shaker for 1 hour at a pressure of 275,792 kPa. The reaction mixture is diluted with diethyl ether (40 ml) and the phases are separated. The organic phase is washed with water (twice 5 ml). The aqueous phases are combined, filtered through a hardened filter paper equal to 52, washed with diethyl ether (2 times 20 ml) and pumped under vacuum.

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%); I.R. (nujol) V max: 1750 (s, A-lactam C = 0) and 1590 cm “^ (s, C = 0); 1 H NMR (DgO) S: 4.25-3.95 (2H, m, H-1 ', H-5), 3.70-3.40 (6H, m, CH2N +), 3.35 (1H, dd, J = 6.1 Hz, J = 2.6 Hz, H-6), 3.08, s, CH3N +), 5.25-2.75 (8H, CH2S, H-4), and 1, 24 ppm (3H, d, J = 6.4 Hz, CH3); U.V. (HgO, c 0.062) Amax: 299 (£ 10.962) T1 / 2: 17.7 h (0.1M phosphate buffer pH 7, 37 ° C).

10 EXAMPLE 17

Preparation of (5R, 6S) -3-L 2- (l-methylmorpholino) · -ethylthio 3 -6- L (R) -l-hydroxyethylJ -7-oxo-1-azabicyclo L 3.2.0 J hept-2- ene-2-carboxylate ÖH

Vr \ /: - N S © N 1 0 "\ CO Θ 2 20 A. 1-Methyl-1- (2-mercapto-ethyl) morpholinium trifluoromethanesulfonate

Me _

25 f + CF3S03H + yS · ^ ^ HSCH2CH2N_jD

| CF3SO30

Me 30 A of N-methylmorpholine (3.29 ml, 0.030 mole)

trifluoromethane sulfonic acid (1.327 ml, 0.015 mole) is added dropwise at 10 ° C, followed by ethylene sulfide (0.89 ml, 0.015 mole). The resulting yellow-brown solution is heated (oil bath) to 50-60 ° C

129 under nitrogen for 18 hours. The volatile material is then removed under vacuum and the residual oil is taken up in 10 ml of t ^ O. The aqueous solution is washed with · diethyl ether (3 times 5 ml) and then the residual organic solvent is removed in vacuo. The resulting aqueous solution is applied to a C 6 g reverse phase column which is eluted with water, then 5% acetonitrile-H 2 O and finally 10% acetonitrile-HgO. Evaporation of the appropriate fractions produces a white solid which is dried under vacuum (P2O5) to give the product (1.92 g, 41%). 10 IR (KBr) v> max: 2560 (-SH) cm- ^ ; NMR (dg-acetone) &: 4.25-3.6 (m, 8H), 3.49 (s, 3H, N-Me), 3.35-2.7 (m, 5H).

B. P-nitrobenzyl diphenylphosphate (5R, 6S) -3- [.2- (1-methylmorpholino) ethylthio ^ -6- [(R) -1-15 hydroxyethyl]] -7-oxo-i-azablcyclo ^ 3.2 .0 ^ hept- 2-ene-2-carboxylate i GH OH - ”iX>" · · jxv 0 '0 \. 0 C02PMB _ C ° 2Ptffi (0O) 2PO® 25 To a solution of p-nitrobenzyl (5R, 6S) -6- [(R) -1-hydroxyethyl "] -3,7-dioxô-1-azabicyclo £ 3.2.0 J heptane-2-carboxylate (0.348 g, .1, -0 mmol) in 25 ml d 'anhydrous acetonitrile on% joust. drip of diisopropylethyl amine 30' (0.191 ml, 1.1 mmol) and then diphenyl chlorophosphate (0.228 ml, 1.1 mmol) at 0 ° C under nitrogen. After stirring at 0 ° C for Ί hour, diisopropylethylamine (0.226 ml, 1.3 mmol) is added to the resulting enol phosphate, followed by the I 130 trifluoromethanesulfonate 1 ~ methyl-1- (2-mercaptoethyl) morpliolinium (0.373 g, 1.2 mmol). The reaction mixture is stirred at room temperature for 1.5 hours and then concentrated in vacuo. The residual material is taken up in water and applied to a C18 reverse phase column. Elution with water, then 20% acetonitrile-H20 and finally 30% acetonitrile-H20 followed by lyophilization of the appropriate fractions gives the product (0.360 g, 40%) in the form of an amorphous solid. I.R. (film) n) 3300 (-0H), 10 1770 (/ 3-lactam CO), 1700 (-C02PNB) cm “1; 1 H NMR (dg-acetone): 8.25, 7.80 (ABq, J = 8.6 Hz, 4H, aromatic), 7.4-6.8 (m, 10 H, diphenylphosphate), 5.56 -5.27 (ABq, J = 14.2 Hz, 2H, benzyl), 4.42 (d of t, J = 9.2 Hz, J '= 2.7 Hz, 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- Π 2- (1 -methylmorpholino) ethylthio 1 -6- 1 · (R) -l-hydroxyethyl j -7-oxo-1 -azabicyclo | _ 3 "2.o] - hept-2-ene-2-carboxylate 20

OH OH

? Me, - X _ Me / - \ o \ 9 o \

25 C02PNB (^ 0) 2Ρ0Θ CO P

To a solution of benzyl p-nitro-3Q diphenylphosphate (5R, 6S) -3-C 2- (1-methylmorpholino) ethylthio ~ \ -6- C (R) - 1-hydroxyethyl J -7-oxo-1- azabicyclo [_3 · 2.0] hept-2-ene-2 carboxylate (0.360 g, 0.49 mmol) in 13 ml of phosphate buffer (0.05 M pH 7.4) 0.36 g of palladium 10 is added % on charcoal, 20 ml of tetrahydrofuran and 20 ml of diethyl ether. This mixture is hydrogenated (Parr) at one! ί 131 pressure of 220.633 kPa for 1 hour. The mixture is filtered through a pad of Celite and the filter pad is washed with H2O and diethyl ether. The aqueous phase is separated and the pH is adjusted to 7.0 with additional phosphate buffer at pH 7.4. After removal of the residual organic solvents under vacuum, the aqueous solution is applied to a C ^ g reverse phase column. Elution with water and freeze-drying of the appropriate fractions produces 0.130 g of an amorphous solid. This material is repurified by reverse phase hplc to give the pure product (0.058 g, 34%) of an amorphous solid.

"IR (KBr) ^: 3420 (br, OH), 1750 (A-lactam CO), 1590 (-C02) cm” '; NMR Ή (D20) d: 4.35-2.77 (m, 17H) , 3.18 (s, 3H, N-Me), 1.23 (d, J = 6.3 Hz, 3H, CHMe); 15 UV (H20) Xmax: 300 (£ 6344Î nm; t ^ (pH 7 , 4, 36.8 ° C) 18.5 h.

EXAMPLE 18

Preparation of (5R> 6s) 3- ^ 2- (1,4-dimethyl-1-piperazinium) ethylthiol -6- L 1 - (R) -hydroxyethyll-2o 7-oxo-1-azabicyclo T. 3.2.0] hept-2-ene-2-carboxylate J. ^ ΑΛ '' \ - / V ς ^ \ "—CH, • I r5 w 25 Λ-n —i

0 cocP

A. 1- (2-acetylthioethyl) -1, -4-dimethyl-piperazinium bromide 9 Br / \ Acetone + N-, Br PH LAv / + - N N— -CH3CS \ _ /

\ / 50 ° C

30 ’X’ 132

A solution of 2-bromoethyl thioacetate (2.20 g, 0.012 mole) and 1,4-dimethylpiperazine (1.95 ml, 0.014 mole) in acetone (4 ml) is stirred at 50 ° C for 65 hours . After cooling to 25 ° C, the liquid phase 5 is decanted from the gum which is. triturated twice in diethyl ether; a powder, yellowish hy coarse copic, 3.2 g (90%) is obtained; I.R. (Nujol) \> ·. 1685 (0 = 0 Λ r * JJlCLX.

thioester) cm "; NMR H (D ~ 0) S: * '-H“ Λ Ά / CH3 10 2.37, 2.39 (2s, 6H, CH ^ O, ^ N-Ci ^), 3, 18 (s, 3H, ^ N).

'ï * B, Hansen, Acta Chem. Scand. 11, 537-40 (1957).

B. 1.4-Dimethvl-1- (2-mercaptoethyl) piperazinium bromide and hydrochloride CH ^ - "Br" V "Br" HC1 3 N_ / A N- '20

A solution of 1- (2-acetylthioethyl) - 1,4-dimethylpiperazinium bromide (1.1 g, 3.7 mmol) in 4 © 6N hydrochloric acid (4 ml) is heated to 80 ° C under nitrogen atmosphere for 1 hour. The solution is concentrated under reduced pressure to give a white powder, 0.41 g (38 #), 1 H NMR (EKSO, dg) S: 2.90 (s, - \ -Λ / +% 30, îC), 3 , 26 (s,>).

-J xch3 ->

Anal, hold, for CgH ^ NgSBrCl.HgO: C 31.03, H 7.16, N 9.05, S 10.35; find ; C 31.62, H 7.46, N 9.19, S 10.19.

! ! 133

(5R, 6s) paranitrobenzyl diphenyiphosphate 3-ί 2 ~ (1> 4-dimethyl-1-piperazinium) -ethylthio -6-ί ^ “(S) -hydroxyethyl 3-7-oxo-1-azat) icyclo [[ 5.2.0] -hept-2-ene-2-carboxylate 5 nM 1> NEt (iPr) p

0H ^ 2 OH

yh — 2) ClP (OPh) „-Λ \ / Γ \,. 8 _ 10 0 § tr ~ N- ~ (

COOPNB \ _ / COOPNB

s 4) NEt (iPr) 2 15 To a cooled solution (0 ° C) of (5 »6s) para nitrobenzyle 6- 11- (R) -hydroxyethyl3 -3,7-d.ioxo-1-aza-bicyclo 3.2 .0d heptane-2- (R) -carboxylate (0.465 g, 1.33 mmol) in acetonitrile (2 ml) maintained under an atmosphere of nitrogen is added diisopropylethylamine 20 (0.278 ml, 1.59 mmol) and diphenyl chlorophosphate (0.33 ml, 1.59 mmol). The reaction mixture is stirred for 30 minutes and treated with a suspension of 1,4-dimethyl-1- (2-mercaptoethyl) -piperazinium hydrochloride (0.40 g, 1.37 mmol) in a mixture of 25 d acetonitrile (3 ml) -water (1 ml) and diisopropylethylamine (0.278 ml, 1.59 mmol). After stirring for 18 hours at 5 ° C, cooled water (15 ml) is added to the mixture. The resulting solution is chromatographed on a PrepPAK-500 / C <d8 column (trade name of Waters 3Q Associates) (2.5 x 7 "5 cm) with as elution solvent 25-359 (acetonitrile in water for give a yellowish powder 0.50 g (50%) after lyophilization; IR (KBr) 0 max: 1765 (C = 0 of fi> -lactam), 1690 (C = 0 of the PNB ester), 1585 (phenyl) , 1512 (N02), 875 (N02) cm "1.35 NMR * H (DMSO, dg) S: 1.16, 1.18 (2d, J = 6.1 Hz, 3H, CH ^ CHOH), 134 CH, ο - \ ~ + \ / 3 ^, 44 (s, _-CH ^), 3.14 (s,), 5.31 (d, J = 6 Hz, OH), 5.39 (center of ABq, J = 13 Hz, CH £ 2 d © PNB), 6.6-7.4 (m, 10H, phosphate phenyl),? ”71 (d, J = 8.8 Hz, 2H, Ho of PNB ), 8.26 (d, J = 8.8 Hz, ftïû of PNB).

(5R, 6S) 3- 2- (1,4-dimethyl-1-piperazinium) ethyl- Ί 10 thio} -6- Çl- (R) -hydroxyethyl -7-oxo-1-aza- bicyclo L3.2.0 Jhept -2-ene-2-carboxylate

OH

X P 10 ° /, 15 Aro ^ / ry

Cr N buffer! _ N // '-'

COOPNB

C00 To a solution of diphenylphosphate of (5R, 6S) -paranitrobenzyl 3- U 2- (1,4-dimethyl-1-piperazinium) ethyl-thio 3 -6- £ 1- (R) -hydroxyethyl3 -7-oxo- 1-azabicyclo j] 3.2.o] [- bept-2-ene-2-carboxylate (0.47 g, 0.623 mmol) in wet tetrahydrofuran (25 ml) add diethyl ether (25 ml), buffer monobasic potassium phosphate-sodium hydroxide (13 ml, pH 7.22) and 10? palladium on charcoal (0.47 g). The resulting mixture is hydrogenated at 23 ° C under a pressure of 275.792 kPa for 1 hour. The two layers are separated and the organic layer 30 is extracted with water (2 times 7 ml). The aqueous layers are combined, filtered through a pad of Celite, washed with diethyl ether (2 times 15 ml) and

Chromatographed on a PrepPAK-500 / C ^ g column (name = Waters Associates commercial) (2.5 x 9.5 cm) with ί 135 water as elution solvent to give 0.097 g (43? 0 after lyophilization ; IR (KBr) Ό max: 3000-3700 (OH), 1750 (C = 0 of P-lactam), 1585 (carboxylate) cnf ^, 1 H NMR (D20) S: 1.24 (d, J = 6 , 4 Hz, 3H, Cï ^ CHOH), 2.33 (s, 3H, 5 _, _ CH, \ - \ / “3 _ / f-CH5), 3.15 (s,) 4.0-4 , 5 (m, H-5, CH3CH0H), UV (Η20) Λ 296 (fc 9476), [«] 3 61.1 ° (C 0.26, HO), - I ^ - ti = 12.4 h (measured at a concentration of 10 "M in 10 3 phosphate buffer pH 7.4 at 36.8 ° C).

EXAMPLE 19

Preparation of (5R, 6S) -3- L2- (N-methyl-thio-morpholinium-oxide) ethylthio ~ 3 -6- Ç1- (R) -hydroxy-15 ethyl] -7-oxo-1-azabicyclo (3.2 .0) hept-2-ene-2-carboxylate

OH

20 'V — sv \

Js— N ^ // N S ~ - ~ 0 r Ί n / v_ / C02® Me

25 OH OH

„® / \ mCPBA 'V-ΤΛ _ © / \ ^ 7οθ Me °' Ig1® '/ 30 2 C02' <3 136 To a cooled solution (-10 ° C) of (5R, 6S) -3- £ 2 - (NHnethyl-thiomorpholinium) ethylthioJ -6- £ l- (R) -hydroxyethyl -7-oxo-1 -azabicyclo (3.2.0) hept-2-ene-2-carhoxylate (608 mg, 1.65 mmol) in a 1: 1 mixture of acetonitrile-water (9 ml), m-chloro-perbenzoic acid (334.8 mg, 1.65 mmol) is added in small portions over a period of one hour. The mixture is then diluted with water (15 ml) and washed with diethyl ether (3 times 15 ml). The aqueous phase is pumped in vacuo and passed through a column of reverse phase silica gel (water) to give a solid which consists of a mixture of compounds. This mixture is separated by reverse phase HPLC and a fraction A 52.4 mg (yield 12%) and a fraction B 23.6 mg (yield 6%) are produced as diastereoisomers of the title compound;

Fraction A: IR (Nujol) max: 1750 (s, / 9-lactam C = 0) and 1580 cm "1 (s, C = 0); 1 H NMR (D £ 0) S: 4.26-2.91 (20H, m, H-4, H-5, H-6, H-1 ', CH2S, CH2S-0, CH3-N + and CH2N +) and 1.24 ppm (3H, d, J = 6.4 Hz , CH3); UV (η £ 0, c 0.0-6) Xmax: 20 302 (^ 10425); T 1/2: 12 h (phosphate buffer 0.065 M, pH 7.4, 37 ° C). Fraction B; ir (Nujol) ^ max: 1750 (s, / 1-lactarne C = 0) and 1585 cm "1 (s, C = 0); 1H NMR (D20) S: 3.86-2.90 (17H, m, H-4, H-5, H-6, H-1 ', CH2S, CH2S-0, CH2N +), 3.25 (3H , a, CH, N +) and 1.24 ppm (3H, 25 d, J = 6.4 Hz, CH3); U.V. (HgO, c 0.05) Λ: 2.99 (8,6517), T 1/2: 10.75 h (buffer solution 0.065 M, ph 7.4, 37 ° C).

137 EXAMPLE 20

Preparation of (5R, 6S) chloride -3-l2- (1,4,4-trimethyl-1-piperazinium) -ethylthio_] -6- [1R-hydroxyethyl] ~ 7-oxo-1-azabicyclo (3.2.0 ) hept-2-5 ene-2-carboxylate

OH

X CH-. / - CH ~ r t \ \ 3 / ¢) 3 (C \

! -T \ _ XN ^, Cl U

ίο 'κ> - s \ _ / ^ ch, ^ -N- ^ 3

0 \ Q

coou A. Bromide and iodide of 1- (2-acetylthioethyl) -1,4,4- 15 trimethylpiperazinium "\ / ~ \ t 8 ^ ν + Λ. (. Βτ-, Γ 20 CH3CS ~ X_J- 'Br' y —OH, 55-60®C \

A suspension of 1- (2-acetylthioethyl) -1,4-dimé thylpiperazinium (1.48 g, 5.0 mmol) in isopropyl alcohol (10 ml) is treated with iodide methyl (0.573 ml, 6.0 mmol) and heated to 55-60 ° C for 50 hours. 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 hot water (8 ml) and the solution is diluted with acetone until turbidity (70-80 ml). Two successive crystallizations give 1.5 g, melting point 220-5 ° C decomposition, 68% of the title compound; I.R. (KBr) Ό max: l692 cm * ”^ (0 = 0); 1H NMR (D20) £: 2.40 (s, 5H, CHjCOO), 5.57 (s, N-CH ^), i: 138! * 3.99 (s); U.V. (HgO) Λ max: 226 (£ 13144). Anal, hold for C11H24N20SBrI: C 30.08, H 5.51, N 6.38; found: C 30.48, H 5.53, N 6.86.

5 B. 1- (2-Mercaptoethyl) -1,4,4- trimethylpiperazinium 10 5 '\ f + V + _ _ HCl 6N Permutit \ f ~~ "„ 4 CH3CS' - ^ J, N_ν ^, βγ, ι g_ï C1r * '2C1 ù 15

A mixture of 1 - (2-acetylthioethyl) -1,4,4-trimethylpiperaziniura iodide bromide (1.84 g, 4.19 mmol) and 6N hydrochloric acid (15 ml) is heated to 57 ° C. under atmosphere nitrogen for 2.5 hours. The solution is concentrated under reduced pressure until dry. The solid is suspended in water (10 ml) and the well-stirred suspension is treated with Permutit S-1 Cl "(trade name) until a solution is obtained.

The solution is poured onto a column (1.2 x 60 cm) of 25 Permutit S-1 Cl “. The column is eluted with water (1.5 ml / min). The appropriate fractions are combined and lyophilized to give a white powder, 0.93 g, m.p. 190-191 ° C, 85%; I.R. (Nujol) 0 raax: 2460 (SH); 1 H NMR (D20) S: 3.4 (s, N-CH3), 3.45 (s, N-CHj, 4.07 (s), 30 Anal, hold for C ^ H ^ N ^ Clg'O ^ S H2O: C 39.34, H 8.62, N 10.20, S 11.67; found: C 39.48, H 8.39, N 10.55, S 11.15.

139 C · (5R, 6S) paranitrobenzyl 3-L 2- (1> ^> 4 ~ trimethyl-1-piperazinium) -ethylthio ~] 6-11R-hydroxyethyl ~] -7-QXQ-1-azabicyclo (3.2 .0) hept- 2-ene-2-carboxylate 5 '1) NEt (iPr) 2

OH 0 OH

VfVn 2) C1P (0Ph) 2 Ρ1Ταί- \ Κπ, 10 ûJ-N- / "3TNEt (iPr) 2" ^ "0JZOs- ^ O \ + (ÎOOPNB \ '\ / _ rnnDMP 2C1”

· :. 4) HS ^^ U \ »2C1 C P

15 To a cooled solution (5 ° C) of (5R, 6S) para-nitrobenzyl 6- [.IR-hydroxyethyl ^ -3,7-dioxo-1-azabicyclo- (3.2.0) heptane-2R-carboxylate (0 , 94 g, 2.7 mmol) in acetonitrile (3 ml) maintained under a nitrogen atmosphere is added diisopropylethylamine (0.557 ml, 3.2 mmol) and diphenyl chlorophosphate (0.663 ml, 3.2 mmoles).

The reaction mixture is stirred at 5 ° C for 30 minutes and treated with diisopropylethylamine (0.599 ml, 3.44 mmol) and an aqueous solution (4 ml) of 1- (2-mercaptoethyl) bischloride -1.4 , 4-trimethylpiperazinium (0.98 g, 3.44 mmol). After 1.25 hours, diisopropylethylamine (0.1 ml, 0.57 mmol) is added and stirring is continued for 2 hours. Part of the acetonitrile is removed under reduced pressure and the resulting red mixture is chromatographed on a PrepPAK-500/30 C ^ Q column (Waters Associates) with 25-75% acetonitrile in water as elution solvent. to give a yellowish powder (1.4 g) after lyophilization. The powder is dissolved in water and the solution is passed through a column (1.2 x 58 cm) of Permutit S-1 Cl ~ using 140 water as the elution solvent. Lyophilization of the appropriate fractions gives 1.17 g of a powder which is repurified on a column of PrepPAK-500 / C.jg. Lyophilization of the appropriate fractions gives a yellowish powder, 0.80 g (53%); IR (KBr) -0 3400 (br, 0H), Λ Πι3 · Χ 1770 (C = 0 du / = -lactame), 1690 (C = 0 ester PNB), 1605 (aromatic), 1515 (N0 £ ) cm "1; 1H NMR (ϋ £ 0) &: 1.26 (d, J = 6.3 Hz, 3H, CH3CH0H), 3.39 (s, NCH3), 4.00 (s), 5, 37 (br, s, CH? _ Of PNB), 7.60 (d, J = 8.6 Hz, 2H, Ho of PNB), 10 8.20 (d, J = 8.7 Hz, 2H, Hm PNB); UV (Η £ 0) λ max: 276 (£ 12094), 306 (£ 10752). Anal, hold for C25H36N4 ° 6SC12 * 3H20: C 46.51, H 6.56, N 8.68, S 4.97,

Cl 10.98; found: C 46.31, H 6.18, N 8.57, S 5.36,

Cl 11.37.

D. (5R, 6S) -3- j ~ 2- (1,4,4-trimethyl-1-piperazinium) ethylthio chloride] | 6- £ 1R-hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

20 OH OH

/ +, 201-- „lMs / - 'Λ1 / w.

COOPNB C00 "Cl 25

Hydrogenate a mixture of (5R, 6S) paranitrobenzyl bischloride 3- £ 2- (1,4,4-trimethyl-1-3q piperazinium) ethylthio] -6-LIR-hydroxyethylJ -7-oxo-1-azabicyclo (3.2 .0) hept-2-ene-2-carboxylate (0.40 g, 0.68 mmol), phosphate buffer (30 ml, 0.05 M, pH 7.0), tetrahydrofuran (10 ml), ether (30 ml) and 10% palladium on charcoal (0.40 g) at 23 ° C under pressure of 241.318 kPa for 1 hour. We separate the two phases. The organic phase is extracted with water (10 ml). The aqueous phases are filtered through a pad of Celite, washed with ether (10 ml), 5 concentrated in vacuo to 10 ml and chromatographed on a PrePAK-500 / C ^ 3 column (2.2 x 11 cm ) with water as the elution solvent to give 70 mg (25%) after lyophilization; I.R. (KBr) 0 max: 3400 (br, OH), ^ 1755 (C = 0 du / 9 -lactam), 1585 (carboxylate) cm “^; 1 H NMR (D20) S: 1.24 (3H, d, J = 6.3 Hz, CHjCHOH), 3.36 (s, NCH3), 3.98 (s); U.V. (HgO) Λ m ^: 296 (8 7987); 1¾¾23 35.9 ° (c, 0.30, HgO), ^ / 2 = 9.8 h (measured at a concentration of 10M in a phosphate buffer pH 7.4, at 36.8 ° C).

Claims (15)

142 1, - Process for the preparation of carbapenem derivatives of the formula R1 2 3 4 5 6 H 5. S-A-R7 N COOR2 10 2 in which R is hydrogen and R is chosen from the group consisting of hydrogen; the following substituted and unsubstituted groups: alkyl, alkenyl and alkynyl, 3 having 1-10 carbon atoms; cycloalkyl and cycloalkylalkyl, having 3-6 carbon atoms in the cycloalkyl nucleus and 1-6 carbon atoms in the alkyl parts; phenyl; aralkyl, aralkenyl and 4 aralkynyl in which the aryl part is phenyl and the aliphatic part has from 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl in which the hetero atom or atoms in the heterocyclic moieties mentioned above are selected from the group consisting of 1-4 oxygen, nitrogen or 6 sulfur atoms and the alkyl parts associated with said heterocyclic parts have 1-6 carbon atoms; 7 in which the substituent or substituents with respect to the radicals mentioned above are chosen from the group consisting of: 143 C 1 -C 6 alkyl group optionally substituted by amino, halogen, hydroxy or carboxyl, halogen -OR3 5 S 3 4 -OCNR ^ R ^ 0 - $ nr3r4 -nr3r4 10 NR3 J. \ r3R4 -so2nr3r4 0 15 -nh'cnr3r4 0 3I! LR ^ CNR - -co2r3 20 = ° 0 -OCR3 -SR3 0 25 4r9 0 H q -SRy II 0 -CN 30 -n3 -OSOJl3 u D 35 -0S02R3 144 3 „4 -NRJS02R -NR5C = NR4 h 5 -NR3C02R4 -no2 ^ in which, compared to the substituents mentioned above ^ “i ,,„ -7. A • O above, the groups R ^ and R are independently chosen from hydrogen; an alkyl, alkenyl and alkynyl group having 1-10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl, having 3-6 carbon atoms in the cycloalkyl nucleus and 1-6 carbon atoms in the alkyl parts; phenyl; aralkyl, aralkenyl and aralkynyl in which the aryl part is phenyl and the aliphatic part has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and hetero-cyclylalkyl in which the hetero atom or atoms in the heterocyclic moieties mentioned above are selected from the group consisting of 1-4 oxygen, nitrogen or sulfur atoms and the alkyl moieties associated with the heterocyclic moieties have 1-6 3-4 carbon atoms, or R and R taken together with the nitrogen to which at least one is bonded may form a heterocyclic ring containing 5-nitrogen or 6 members; R ^ is such that • Z. defined for except that it cannot be hydrogen; A O or in which R and R taken together represent a C 2 -C 6 alkylidene radical or C 2 -C 30 alkylidene radical substituted by hydroxy; A is a cyclopentylene, cyclohexylene or C 2 -C 6 alkylene radical optionally substituted by one or more C 1 -C 4 alkyl groups; R ^ is hydrogen, an anionic charge or a readily removable conventional carboxy protecting group, û A 35. the condition that when R ^ is hydrogen or a protecting group 145, a counter anion is also present; 14 and R is an aromatic or non-aromatic heterocycle containing nitrogen in quaternary form linked to A by a nitrogen of the nucleus, thus forming a quaternary ammonium group, or a pharmaceutically acceptable salt thereof, said process being characterized in that it includes the reaction of the intermediate of the formula: ,, TT _8 " 10 R - "H-V'V1 X-N-2 '0 ^ COOR IV IV 18. in which R and R are as defined above, 2 ′ R is a conventional easily detachable carboxyl protecting group and L is a conventional detachable group, with a thiol compound of the formula: HS-A-R1 ^ X® 25 VII 14 in which A and R are as defined above and χθ is a counter anion, in an inert solvent and in the presence of a base, to produce a carbapenem product 30 of the formula: t,! .. 146 _8 „R _ 14„ / Sar, h · r — ΓΥ *, - N- \ 2, 5 0 ^ COOR ^ 1 '* N% in which r \ R®,, A, R ^ and X® are such that; previously defined and, if desired, removal of the 2 'carboxyl protecting group R to give the corresponding deblocked compound 10 of formula I, or a pharmaceutically acceptable salt thereof. 2.- Method according to claim 1, characterized in that the base is a non-nucleophilic tertiary amine or a tri (C <j-C ^) alkylamine. 3. A method according to claim 2, characterized in that the base is a non-nucleophilic tertiary amine. 4, - Method according to claim 1, 2 or 3, characterized in that the reaction is carried out at a temperature of -15 ° C to room temperature. 5. A method according to claim 4, characterized in that the reaction is carried out at a temperature in the range from about -15 ° C to + 15 ° C. 6.- Method according to one of claims 1, 2, 3, 4 or 5, characterized in that the inert solvent is chosen from the group consisting of acetonitrile, of a mixture of acetonitrile, of dimethylformamide, of tetrahydrofuran, a mixture of tetrahydrofuran and water, a mixture of acetonitrile and water and acetone. v- 7.- New synthesis intermediates, characterized in that it is a quaternary thiol amine compound of the formula 147 HS - A - R14 Xe VII in which A is a bivalent cyclopentylene, 5 cyclohexylene or R10 R12 .. II - CC— <. R11 R13 10 ΛΛ 1? 1 ^ in which R, R, R and Ru are each independently hydrogen or a C 1 -C 6 alkyl group, X® 14. h is a counter anion and R represents a mono aromatic or non-aromatic heterocyclic radical , substituted or unsubstituted bi- or polycyclic containing at least one nitrogen in the nucleus and linked to A by a nitrogen in the nucleus, thus forming a quaternary ammonium group. 8.- Process for the preparation of the new synthesis intermediate according to claim 7, constituted by a thiol quaternary amine compound of formula 14 20 HS - A - R ^ m in which A is a bivalent cyclopentylene, cyclohexylene or R10 R12 i I —CC— 25 in i “** in which R ^, R ^, r” '2 and R ^ are each inde while hydrogen or a CL-C alkyl group. ,> ο, λ A 1 ^ = λτ is a counter anion and R represents a heterocyclic aromatic or non-aromatic mono, bi- or substituted or unsubstituted 148 radical containing at least one nitrogen in the nucleus and linked to A by a nitrogen of the nucleus, thereby forming a quaternary ammonium group, said process being characterized in that it comprises the reaction 5 of a sulphide of the formula 6 · <5 - p10 s p12 w, o A ”1“ 10 11 1? 17 in which R, R, R and R ^ are each independently hydrogen or a C ^ -C ^ alkyl group, with a strong acid and either (a) a heteroaromatic amine of the formula, s O in which: O represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocyclic radical containing at least one nitrogen in the nucleus capable of being transformed into quaternary form and linked to a carbon atom of the substi-i * killing A, or ( b) a heterocyclic amine of the formula: 149 R16 in which: \ ”q represents a substituted or unsubstituted mono-, bi- or polycyclic non-aromatic heterocyclic radical containing at least one nitrogen in the nucleus capable of being transformed if \ sous quaternary form through a substituent R and linked to a carbon atom of the substituent A and R represents either (a) an aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, aryl, araliphatic, heteroaryl, heteroaraliphatic, heterocyclic or heterocyclyl radical optionally substituted aliphatic either (b) a divalent C 1 -C 4 alkylene group linked to the ring '' O so as to form a bridged polycyclic group. 9. A method according to claim 8, characterized in that the reaction is carried out at a temperature between about -20 ° C and about 100 ° C. 10, - Method according to claim 8 or 9, characterized in that the strong acid is hydrochloric acid, hydrobromic, methanesulfonic, p-toluenesulfonic or trifluoromethanesulf onic. 11. Method according to one of claims 8 to 10, characterized in that the reaction is carried out in the presence of a non-polar organic solvent, preferably methylene chloride, benzene, xylene or toluene. 150 12, - Method according to one of claims 8 to 11, characterized in that the amine and sulfide reagents are liquid or when the amine reagent is a solid, the sulfide reagent is liquid and the solid amine is soluble in the reagent liquid sulfide, the reaction is carried out without additional solvent. "Vv 'U