CS247195B2 - Method of carbapenem derivatives production - Google Patents

Abstract

Process for producing carbapenem derivatives of formula I, wherein the general symbols have the following meanings and their meanings pharmaceutically acceptable salts by reacting the intermediate of formula II wherein the general symbols have as defined below, with an alkylating agent general formula R5 - X ', wherein the general symbols are as set forth below meanings to form a general compound of formula I ', wherein the general symbols have further the above meanings and, where appropriate, will be replaced anion X 'to another anion and optionally is removed carboxyl-protecting group R2 optionally, compounds of formula I are converted to pharmaceutically acceptable salts. The compounds of formula I are useful to treat bacterial infections.

Description

A process for the preparation of carbapenem derivatives of the formula I wherein the symbols have the following meanings and their pharmaceutically acceptable salts, by reacting an intermediate of the formula II wherein the symbols have the following meanings with an alkylating agent of the formula

R ⁵ - X ', where the symbols are as defined below, to form a compound of the formula I', wherein the symbols are as defined below, optionally replacing the anion X 'with another anion and optionally removing the carboxyl protecting group R 2 and optionally the compounds of formula I are converted into pharmaceutically acceptable salts.

The compounds of formula I are useful for the treatment of bacterial infections.

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

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

-S-A · 'W R

NR ⁶ where

A is C1-C6 alkylene with straight or branched chain, ς

R is an optionally substituted aliphatic, cycloaliphatic, cycloaliphatic-aliphatic, aryl, arylaliphatic, heteroaryl, heteroarylaliphatic, heterocyclic or heterooyl-aliphatic residue and the cycle of the formula represents a nitrogen-containing aromatic heterocycle attached to alkylene 5 group A by a ring carbon atom and quaternized with a substituent.

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

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

The parent carbapenem compounds were natural products such as the thienamycin formula

SCH ₂ CH ₂ NH ₂

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

Other natural carbapenem-containing compounds include olivanoic acid derivatives such as the antibiotic MM 13902 of formula ch ₃

-sch = chnhcoch ₃

COOH claimed in U.S. Patent No. 4,113,856, the antibiotic MM 17880 of the formula

CH, claimed in U.S. Patent 4, claimed in U.S. Pat

HO ₃ S0 sch ₂ ch ₂ nhcoch ₃

COOH

162,304, the antibiotic MM 4550A of the formula

HOjSO

CH and S-CH ₃ ⁰ chnhcoch COOH

172,129, and the antibiotic 890Ag of formula

CH,

NO

II ^ sch = chnhcch ₃

COOH claimed in U.S. Patent No. 4,264,735.

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

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

^{s <0) n} w ^{NHCOCH 3}

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

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

sch ₂ ch ₂ nh

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

SCH ₂ CH ₂ NHCOCH ₃

COOH t

from Streptomyces fermentation medium.

Carbapenems, which are unsubstituted at the 6-position, were also synthesized. Thus, U.S. Pat. No. 4,210,661 discloses compounds of formula s -r ₂

COOH wherein R ₂ is phenyl or substituted phenyl, U.S. Pat. No. 4,267,177 discloses compounds of formula

s-n,

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

S - CR ₂ ⁼ CR 2 R q

0 =

COOH wherein R ₂ and R 8 are hydrogen or alkyl and R 4 -substituted phenyl and n is 1 or 2, and US Patent No. NH-CO 4 R 8, wherein R 8 is alkyl, phenyl or subst. ₁ R ₂

COOH where R is hydrogen and R ₂ is cyano or CO ₂ R g, wherein R 8 is hydrogen, alkyl, aryl or arylalkyl.

Carbapenems of general formula

R @ 1 NH - t

COOH

Wherein R is hydrogen or acyl and R is hydrogen or substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, alkylcycloalkyl, aryl, arylalkyl, arylalkenyl, arylalkinyl, heteroaryl, heteroarylalkyl, heterocyclyl or heterocyclylalkyl are disclosed in U.S. Pat. No. 4,218,463. There are no claims for any heteroarylalkyl

wherein A is alkylene and the moiety n? -r ⁵ is a quaternary nitrogen containing an aromatic rieterocycle attached to the alkylene group A via a carbon atom in the cycle.

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

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

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

OH

O CO ₂ pNB where pNB is p-nitrobenzyl.

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

1) N-formimidoylthienamycin of formula

OH sch ₂ ch ₂ n = c-nh ₂

COOH is exemplified in European Patent Application Publication No. 6,639

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

O

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

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

OH sch ₂ ch ₂ n = cCOOH

Wherein X and Y are hydrogen, R, OR, SR or NRR wherein R is substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl, hetero12 arylalkyl, heterocyclyl or heterocyclylalkyl and R and R is hydrogen or R is exemplified in U.S. Patent No. 4,194,047,

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

COOH

Wherein R is aryl, alkyl, acyl or arylalkyl and R and R are independently selected from hydrogen and acyl (including acyl type

O

-C-R wherein R ¹¹ may be, inter alia, alkyl substituted with a quaternary ammonium group, for example

O řl ©

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

5) compounds of formula

OR ³

O COOH

Wherein R is hydrogen, acyl or a monovalent optionally substituted hydrocarbon radical, R is an optionally substituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkenylalkyl, cycloalkylalkyl, aryl, arylalkyl, heteroaryl or heteroarylalkyl and R is acyl (including acyl type)

O ii

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

O ιι e

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

6) compounds of the general formula

OH ₂ NR ⁵ R ⁶ R ⁷

AND

Wherein R, R and R are independently selected from hydrogen, substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkenylalkyl, cycloalkylalkyl, aryl, U.S. Patent 4,235,920,

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

OR ³ o

R ⁵ t ®, ₂

SCH ₂ CH ₂ NC = NR ¹ R 1 ^!

1 «cox

Wherein R and R independently of the others are radicals of the type defined for R, hydrogen or nitro, hydroxyl, alkoxy of 1 to 6 carbon atoms, amino, alkylamino of 1 to 6 carbon atoms, dialkylamino or trialkylamino of 1 to 6 carbon atoms 1 2 in each of the alkyls and the other anion present in the latter case or R and R may be taken together to form together with the nitrogen atom to which they are attached a substituted or unsubstituted monocyclic or bicyclic heteroaryl or heterocyclic radical containing 4 to 10 ring atoms, one or more of which may be other heteroatoms selected from oxygen, sulfur and nitrogen, R is cyano or substituted or unsubstituted carbamoyl, carboxyl, alkoxycarbonyl of 1 to 11 carbon atoms, alkyl of 1 to 10 carbon atoms, alkenyl of 2 to 10 "Carbon atoms, C 2 -C 10 alkynyl, C 3 -C 10 cycloalkyl, C 4 -C 12 cycloalkylalkyl" carbon atoms, cycloalkylalkenyl of 5 to 12 carbon atoms, cycloalkenyl of 3 to 10 carbon atoms, cycloalkenylalkenyl of 5 to 12 carbon atoms, cycloalkenyl alkyl of 4 to 12 carbon atoms, aryl of 6 to 10 carbon atoms, arylalkyl of 7 to 16 atoms or a monocyclic or bicyclic heteroaryl, heteroarylalkyl, heterocyclyl or heterocyclylalkyl of 4 to 10 ring atoms, of which one or more are heteroatoms selected from oxygen, sulfur and and wherein the alkyl moiety of the heteroarylalkyl or heterocyclylalkyl moiety comprises

2 of from 1 to 6 carbon atoms, the substituent or substituents on R, R, R or the ring formed by 12 R and R are chlorine, bromine, iodine, fluorine, azido, alkyl of 1 to 4 carbon atoms, mercapto, sulfo, phosphono, cyanthio (-SCN), nitro, cyano, amino, hydrazino, substituted amino or hydrazino of up to three alkyls of 1 to 6 carbon atoms, hydroxyl, alkoxy of 1 to 6 carbon atoms, alkylthio of 1 to 6 carbon atoms, carboxyl , oxo, C 1 -C 6 alkoxycarbonyl, C 2 -C 10 acyloxy, carbamoyl, C 1 -C 4 alkylcarbamoyl or C 1 -C 4 dialkylcarbamoyl, R ₇ is hydrogen 44, acyl or a radical of the type defined for R, R is C1-C10 alkyl, substituted carbonylmethyl, C1-C6 alkoxyalkyl in both parts of the substituent, C3-C6 cycloalkoxyalkyl and cycloalkoxy; up to 6 carbon atoms in the alkyl, alkanoyloxyalkyl of 2 to 12 carbon atoms, partially or fully halogenated alkyl of 1 to 6 carbon atoms, wherein the halogen atoms are chlorine, bromine or fluorine, aminoalkyl, alkenyl or alkynyl of 2 to 10 carbon atoms, acyl C 3 -C 14 alkoxycarbonylalkyl, C 4 -C 21 dialkylaminoacetoxyalkyl, C 2 -C 13 alkanoylaminoalkyl, C 1 -C 3 arylalkyl and wherein the aryl radical contains from 6 to 10 carbon atoms, monocyclic or bicyclic heteroarylalkyl or a heterocyclylalkyl having 4 to 10 ring atoms, up to 3 carbon atoms in the alkyl moiety and 1 to 4 heteroatoms selected from oxygen, sulfur and / or nitrogen atoms, a nucleosubstituted arylalkyl or heteroarylalkyl, wherein the substituent is a chlorine, fluorine, bromine or iodine or alkyl of 1 to 6 carbon atoms, aryl or aryl substituted on the core of 6 to 10 carbon atoms and wherein any substituent on the core is hydroxyl yl, alkyl of 1 to 6 carbon atoms, chlorine, fluorine or bromine, arylalkoxylalkyl, alkylthioalkyl of 2 to 12 carbon atoms, cycloalkylthioalkyl of 4 to 12 carbon atoms, acylthioalkyl of 2 to 10 carbon atoms in the acyl moiety and 1 to 6 atoms alkyl in the alkyl or phenylalkenyl wherein alkenyl has 2 to 6 carbon atoms, R is substituted or unsubstituted alkyl of 1 to 10 carbon atoms, alkenyl of 2 to 10 carbon atoms or alkynyl of 2 to 10 carbon atoms, ring substituted or unsubstituted cycloalkyl , cycloalkenyl, cycloalkenylalkyl and cycloalkylalkyl of 3 to 6 carbon atoms and up to 6 carbon atoms in any chain, aryl of 6 to 10 carbon atoms, arylalkyl of 6 to 10 carbon atoms and 1 to 6 carbon atoms in the alkyl moiety, monocyclic or bicyclic heteroaryl or heteroarylalkyl having 4 to 10 carbon atoms in the ring, one or more of which is oxygen, nitrogen or sulfur and 1 to 6 carbon atoms in the alkyl moiety and ring substituents u or on the chain are chlorine, bromine, iodine, fluorine, azldo, cyano, amino, C 1 -C 6 alkylamino, C 1 -C 6 dialkylamino in each alkyl or C 1 -C 6 trialkylamino in each of anion, hydroxyl, C 1 -C 6 alkoxy, C 1 -C 6 alkylthioalkyl, carboxyl, oxo, C 1 -C 6 alkoxycarbonyl, C 2 -C 6 acyloxy are also present in the latter case 10 carbon atoms, carbamoyl, C 1 -C 4 alkylcarbamoyl, C 1 -C 4 dialkylcarbamoyl in each of the alkyls, cyanothio (-SCN) or nitro, R ® is hydrogen, hydroxyl, mercapto, R, -OR, - SR, or 12 12

-NR R where R, R and R are as defined above;

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

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

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

M is a pharmaceutically acceptable cation and

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

8) compounds of the general formula

OH sch ₂ ch ₂ w COO where

R 8 (j attached to the nitrogen atom of the amino group of the thienamycin represents a mono- or polycyclic nitrogen atom containing a heterocyclic group and

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

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

Among the compounds listed in U.S. Pat. No. 4,235,920 are compounds of formula © FCJ SCH ₂ CH ₂ N (CH _{3) 3} and

O COOH wherein A is a pharmaceutically acceptable anion.

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

In addition to the above, carbapenem derivatives also have a wide range of substituents at the 6-position

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

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

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

COOH where optionally substituted pyrimidinyl and R ₂ is CR 1 R ₃ , where R ₃ is hydrogen or hydroxyl, and R 3 is hydrogen, alkyl, benzyl or phenyl or a ring, is hydrogen or R 4 is hydrogen or alkyl and Rq together form carbocyclic

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

R ⁶

R ⁷

SR ⁸

O

COOH

Wherein R °, R and R ° are independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, alkenyl or alkynyl of 1 to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl of 3 to 6 carbon atoms in the cycloalkyl ring having 1 to 6 carbon atoms in the alkyl moiety, aryl such as phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein the substituent or substituents related the above-mentioned residues are selected from the group consisting of -X ° halogen (chlorine, bromine, fluorine), -OH hydroxyl, -OR * alkoxy, aryloxyl O

M 12

-OCNR R is carbamoyloxy O

- <!! nr1r2 Carbamoyl 1 2

-NR R is amino, amidino

-NO ₂ nitro, © 1 1

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

2 ¹ 2.

-C = NOR of oximino

-SR ⁴ alkyl- and arylthio 1 2

-SO 2 NR R R sulfonamido O ¹¹ 1 2

-NHCNR R ureido O

1 ¹¹ 2

-R CNR - amido

-CO 3 H carboxy

-CO ₂ R ⁴ carboxylate O ¹¹ 1

-CR acyl O # 1

-OCR acyloxy

-SH mercapto group O

-R ⁴ alkyl- and arylsulfinyl O

II 1

-SR alkyl- and arylsulfonyl II O

-CN cyano

-Nj azido wherein, with respect to the above-mentioned list of substituents on R, R? and R ® are substituents and R ® is independently selected from the group consisting of hydrogen, alkyl, alkenyl and alkynyl of 1 to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl of 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl portion of the molecule, aryl such as phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl portion is phenyl and the aliphatic portion has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl and wherein the heteroatom or heteroatoms in the above heterocyclic portions they are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and wherein the alkyl moieties associated with these heterocyclic moieties have 1 to 6 carbon atoms (see also European Patent Application Publication No. 1,627, 1,628, 10,317, 17,992, 37,080, 37,081 and 37,082).

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

7 ' ^ch 3 ^{ch s} ' ¹²

J - N - X.

what ₂ H wherein R ¹ is hydrogen or a group selected from the substituents consisting of OH, OSO-H, or a salt or alkyl ester having 1-4 carbon atoms, OR ^2, SR ^3, OCOR ^2, OCO ₉ R ³ or OCONHR ^3, wherein R ²³ is alkyl of 1 to 6 carbon atoms or optionally substituted benzyl and R is alkyl of 1 to 6 carbon atoms or optionally substituted phenyl or benzyl and R is alkyl of 1 to 6 carbon atoms, alkenyl of 2 to 6 atoms C 3 -C 6 alkynyl wherein the triple bond is not present on the carbon atom attached to the sulfur atom, arylalkyl, C 1 -C 6 alkanoyl, arylalkanoyl, aryloxyalkanoyl or arylcarbonyl, each of which R groups are optionally substituted.

European Patent Application no. 44170 demonstrates carbapenem derivatives of formula <°> ni nec SXN .3CO N H ₂ wherein R is hydrogen or an organic group bonded via a carbon atom to the carbapenem ring, n is 0 or 1, X is a saturated or an unsaturated hydrocarbon radical optionally substituted by a bromine or chlorine atom and R is alkyl of 1 to 6 carbon atoms, alkenyl of 2 to 6 carbon atoms, arylalkyl of 1 to 10 carbon atoms or aryl, wherein any of the R ⁴ groups is optionally substituted. However, none of the compounds wherein the tetrazole ring is bonded to X via a quaternized nitrogen atom, i.e. a positively charged nitrogen atom that is not attached to a hydrogen atom, is claimed in the application.

European Published Patent Application No. 38,869 mentioned above discloses syntheses of carbapenem derivatives via intermediates of the general formula wherein R ⁶ R 'and R are as defined above, and is an easily removable carboxyl protecting group. Likewise, compounds of the formula are also shown as intermediates

R ⁶

X

2 *

CO ₂ R ^z wherein X is described as leaving; group.

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

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

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

NR ¹

II -, SAC-NFVR ²

COOH

Wherein R and R are, inter alia, independently selected from the group consisting of hydrogen, alkyl, alkenyl, aryl and arylalkyl, A is a direct single bond linking sulfur and carbon atoms, or A is a cyclic or acyclic linking group selected from among other substituents 1; R 2 and R 2 are alkyl, cycloalkyl, aryl, heteroaryl, or heteroalkyl, R and R which define the carbamidoidoyl function are, inter alia, independently selected from the group consisting of hydrogen, alkyl and aryl; The present invention further provides carbamimidinium residues obtained by quaternizing one of the carbamimidoyl nitrogen atoms. On page 12 of this application, substituents at the 2-position of the group δ are possible

NR ¹ '',

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

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

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

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

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

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

-S-CH, CH, N-CH ₃ was mentioned, as well as carbapen with a quaternized heteroaromatic ring bonded directly to the sulfur substituent.

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

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

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

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

The present invention relates to a process for the preparation of carbapenem derivatives of the general formula I

where

Q

R is hydrogen and

R ³ is selected from the group consisting of hydrogen, substituted or unsubstituted: alkyl, alkenyl and alkynyl of up to 10 carbon atoms, cycloalkyl and cycloalkylalkyl of 3 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the cycloalkyl ring and 1 to 6 carbon atoms in the alkyl moiety, phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein heteroaryl is a mono-, di- or polycyclic aromatic heterocyclic radical with 5 or 6 members in each ring and heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic heterocyclic residue with 5 or 6 members in each ring and wherein the heteroatom or heteroatoms in the above-mentioned moieties are selected from the group consisting of 1 to 4 atoms oxygen, nitrogen or sulfur, and the alkyl moieties attached to said heterocyclic moieties have an alkyl moiety; (C 1 -C 6) wherein the substituent (s) of the abovementioned radicals are independently selected from the group consisting of C 1 -C 6 alkyl optionally substituted with amino, halogen, hydroxyl or carboxyl, halogen

-OR ³

O

IR 3 4 -OCNR R

4 -NR R

NR

3 '4

-so ₂ nr ^j r * * 3 4

-NHCNR R

The four -R CNR

-OP (O) (OR ³⁾ (OR ^{4 * * * *)} = 0

M 3 -OCR

-SR

O

-ŠR ⁹

-CN

-N

-oso ₃ y

About «

-OS ·> 1 o

, 9

3 «

-NR S-R

O

-NR C-NR k ³

4

-NR CO ₂ R

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

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

-OCO ₂ R ³

-OCOR ³

-contr ³ r ⁴ o

H ₉ -OS-R 1

O

4 -NR R

4

R ^J C0NR 3 3

-NR CO ₂ R

31 q -NR ^J SR ^y

II

O

-SR

O 0

-V- R ⁹

-SO ₃ ^r3

-what ₂ r ³

4

-CONR R

-CN phenyl optionally substituted with 1 to 3 fluorine, chlorine, bromine, alkyl of 1 to 6 carbon atoms, OR 1 -HR 2 R 4, -SO 2 R 4, -CO 2 R 4, or -CONR 4 R 4, wherein R 6 and R 6 in the substituents R 6 are as defined above, or R 0 may be attached elsewhere in the ring to form a fused heterocyclic or heteroaromatic ring, which may further comprise additional heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur, is hydrogen, substituted and unsubstituted alkyl, alkenyl and alkynyl of 1 to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl of 3 to 6 carbon atoms in the cycloalkyl ring of 1 to 6 carbon atoms in the alkyl moieties, spirocycloalkyl of 3 to 6 carbon atoms, phenyl, arylalkyl, arylalkenyl and arylalkinyl wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, heterocyclyl and heterocyclylalkyl, wherein the heteroaryl is a mono-, di- or polycyclic aromatic heterocycle a 5-membered or 6-membered heterocyclic moiety and the heterocyclyl is a mono-, di- or polycyclic saturated or unsaturated non-aromatic 5-membered or 6-membered heterocyclic moiety and wherein the heteroatom or heteroatoms in the above heterocyclic moieties are selected from the group consisting of up to 4 oxygen, nitrogen or sulfur atoms, and the alkyl moieties attached to these heterocyclic moieties, have 1 to 6 carbon atoms, wherein the substituent or substituents of the above moieties are selected from the group consisting of amino, mono-, dia trialkylamino, hydroxyl, alkoxy, mercapto , alkylthio, phenylthio, sulfamoyl, amidino, guanidino, nitro, chlorobromo, fluoro, cyano and carboxyl, and wherein the alkyl moieties in the above substituents have 1 to 6 carbon atoms,

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

R is a hydrogen atom, an anionic charge, or a conventional carboxyl protecting group which is easily cleavable, selected from the group consisting of benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloroethyl, trialkylsilyl wherein alkyls have 1 to 4 carbon atoms, phenacyl, methoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridiylmethyl, C1-C6 alkyl, pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, methoxymethyl, and preferably p-nitrobenzyl, wherein when R is hydrogen or a protecting group, an ion is also present and the group represents a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocyclic radical containing at least one ring nitrogen atom and a total of 1 to 4 nitrogen, oxygen or sulfur atoms and attached to A through a ring carbon atom and having a ring nitrogen atom which is quaternized with R 1 and pharmaceutically acceptable salts thereof, characterized in that the intermediate compound is reacted formula II

wherein R1, R8, _and ^R15 are as defined above. '

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

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

S-A z- ©

COOR ² '' 15

N-r5, as defined above, and optionally replacing anion X with another anion, and optionally removing the 2 'carboxyl protecting group R to form an unprotected compound of formula I or pharmaceutically acceptable salts thereof.

Compounds of formula X are potent antibacterial agents or intermediates useful in the preparation of these agents.

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

The novel compounds of formula I above contain a carbapenem core

and may be referred to as 1-carb-2-penem-3-carboxylic acid derivatives. Alternatively, the compounds may be considered as compounds of the formula

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

The compounds of formula I may be unsubstituted at the 6-position or substituted by the 8 L groups mentioned above for the other carbapenem derivatives. More specifically, R can be a hydrogen atom and R can be a hydrogen atom or a substituent other than a hydrogen atom, as disclosed, for example, in European Patent Application Publication No. 38,869 (see definition of Rg).

1

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

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

More specifically, R may be a hydrogen atom or a substituent other than the hydrogen atom shown in the example given in European Published Patent Application No. 54,917 (as defined by R or R) or in U.S. Patent No. 4,350,631.

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

15 Dec

For definitions for R, R and R expressions

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

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

b) heteroaryl includes mono-, bi- and polycyclic aromatic heterocyclic groups having 1 to 4 oxygen, nitrogen or sulfur atoms, preferably 5- or 6-membered heterocyclic rings such as thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, fetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyrilazinyl, pyrrolyl, pyrazolyl etc .;

c) heterocyclyl includes mono-, bi- and polycyclic saturated or unsaturated non-aromatic heterocyclic groups containing from 1 to 4 oxygen, nitrogen or sulfur atoms, preferably 5- or 6-membered heterocyclic rings such as morpholinyl, piperazinyl, piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrvolinyl, pyrrolidinyl and the like;

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

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

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

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

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

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

The compounds of formula X in the form of an acid salt may be written as

R = H or a protecting group wherein x 'is an acid anion. Anion x may be selected to provide a pharmaceutically acceptable salt for therapeutic applications, but in the case of intermediates of compounds of formula I, anion X may also be a toxic anion.

In this case, the ion can then be removed or substituted with a pharmaceutically acceptable anion to form an active end product for therapeutic use. When acidic or basic groups are present in or R 1 groups or in the residue

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

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

Preferred compounds of the present invention are compounds of formula I wherein R is hydrogen and R 6 is hydrogen, CH 2 Cl 2 ^- ,

CH.

'CH-,

CH. OH \ t

CH.

OH or CH ₃ CH 2 of this subgroup are preferred compounds wherein R is

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

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

CH. '

The alkylene moiety (i.e., substituent A ') in the compound of formula I may be straight or branched chain and may contain 1 to 6 carbon atoms. Preferred compounds are those wherein A ^'(CH 2 ^ n ", ^wherein n is 1 or 2 and particularly preferred embodiment comprises those compounds

The alkylene moiety of A is attached via a ring carbon atom to the N-substituted quaternized aromatic heterocycle of the formula wherein the r 5 substituent is preferably optionally substituted alkyl of 1 to 6 carbon atoms, alkenyl of 2 to 10 carbon atoms, alkynyl of 2 to 10 atoms carbon, cycloalkyl of 3 to 6 carbon atoms, cycloalkylalkyl of 6 carbon atoms in the cycloalkyl and 1 to 6 carbon atoms in the alkyl part, phenyl, phenylalkyl of 1 to 6 carbon atoms in the alkyl part, phenylalkenyl of 2 to 6 carbon atoms in the alkenyl part phenylalkynyl having 2 to 6 carbon atoms in the alkynyl moiety, heteroaryl, heteroarylalkyl wherein the alkyl moiety has 1 to 6 carbon atoms, heterocyclyl or heterooyl alkyl wherein the alkyl moiety has 1 to 6 carbon atoms.

The heteroaryl (or heteroaryl portion of heteroarylalkyl) as the R substituent may be a mono-, bi-, or polycyclic, aromatic heterocyclic group containing from 1 to 4 oxygen, nitrogen or sulfur atoms, preferably a 5- or 6-membered heterocyclic ring such as thienyl, furyl, thiadiazolyl, oxadiazolyl, triazolyl, isothiazolyl, thiazolyl, imidazolyl, isoxazolyl, tetrazolyl, oxazolyl, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl and pyrazolyl.

C

The heterocyclyl (or heterocyclic moiety of the heterooyklylalkyl) as the R substituent may be a mono-, bi- or polycyclic saturated or unsaturated non-aromatic heterocyclic group containing 1 to 4 oxygen, nitrogen or sulfur atoms, preferably 5- or 6-membered heterocyclic rings such as morpholinyl, piperazinyl , piperidyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, pyrrolinyl and pyrrolidinyl.

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

(a) alkyl of 1 to 6 carbon atoms, optionally substituted with 1 to 3 substituents selected from the group consisting of amino, fluoro, chloro, carboxyl, hydroxyl or carbamoyl;

b) fluorine, chlorine or bromine,

o) -QR ³

(d) -OCO ₂ R ³ ;

e) -OCOR ³ ,

f) -OCONR ³ R ⁴ , if q

g) -OS-R;

(h) -oxo,

4

(i) -NR R,

j) r ³ conr ⁴ -,

k) -nr ³ co ₂ r ⁴

(l) -NR ³ CONR ³ R ⁴

O ll q

m) -NR S-R 11;

O

n) -SR ³ ,

o) -SOR ⁹ ,

O ¹¹ 9

p) -S-R,

(q) -SO ₃ R ³

(r) -CO ₂ R ³ ,

s) -CONR ³ R ⁴

t) -CN or

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

-NR R, -SO 2 R, -CC 1 R or -CONR R, wherein, with respect to the aforementioned substituents r 5, R 4 and R 4 are independently selected from the group consisting of hydrogen, alkyl, alkenyl, and alkynyl of 1; up to 10 carbon atoms, cycloalkyl, cycloalkylalkyl and alkylcycloalkyl of 3 to 6 carbon atoms in the cycloalkyl moiety and 1 to 6 carbon atoms in the alkyl moiety, phenyl, arylalkyl, arylalkenyl and arylalkinyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1 to 6 carbon atoms, heteroaryl, heteroarylalkyl, hetero cyclyl and heterocyclylalkyl, wherein the heteroaryl and heterocyclyl group or moiety is as defined above for R ° and the alkyl moieties attached to these hetero-3 cyclic moieties have 1 to 6 carbon atoms, or R and R together with the nitrogen atom to which at least one is attached may form a five- or six-membered nitrogen containing a heterocyclic (as defined above for R) ring and R is defined as above except that it is not a hydrogen atom. Most preferred substituent

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

In addition, the R substituent together with another atom in the ring portion of the molecule may form a fused heterocyclic or heteroaromatic ring, which ring may contain additional preferably 1 or 2 heteroatoms selected from oxygen, nitrogen and sulfur.

For example

Group

or preferably represents a substituted or unsubstituted mono-, bi-, or polycyclic aro 247175 heterocycle containing at least one ring nitrogen atom and 0 to 5 additional heteroatoms selected from oxygen, nitrogen and sulfur, the heterocyclic ring being attached to A through a ring carbon atom and has a quaternary ring nitrogen atom.

The heteroaromatic β-N ring may be optionally substituted on available ring carbon atoms, preferably 1 to 5 substituents, preferably 1 to 3 substituents independently selected from the group consisting of C 1 -C 4 alkyl, C 1 -C 4 alkyl preferably substituted by 1 to 3 hydroxy, amino, (C 1 -C 4) alkylamino, (C 1 -C 4) dialkylamino (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy, carboxy, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, bromine or fluorine) or sulfo groups, cycloalkyl of 3 to 6 carbon atoms, cycloalkylalkyl of 3 to 6 carbon atoms in the cycloalkyl moiety and 1 to 4 carbon atoms in the alkyl moiety, optionally substituted with 1 to 3 substituents. above in connection with alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, alkylthio of 1 to 4 carbon atoms, amino (C 1 -C 4) dialkylamino, halogen, (C 1 -C 4) alkanoylamino, (C 1 -C 4) alkanoyloxy, carboxyl, sulfo, -C 1 -C 4 -alkyl, hydroxy, amidino, guanidino , phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group consisting of amino, halogen, hydroxyl, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, alkylamino of 1 to 4 carbon atoms, dialkylamino having 1 to 4 carbon atoms in each of alkyl, carboxy, and sulfo; C 1 -C 4 phenylalkyl, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl, and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above with respect to alkyl of 1 to 4 carbon atoms, and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl moiety associated with the heteroarylalkyl has 1 to 6 carbon atoms, the heteroaryl and heteroarylalkyl groups being optionally substituted in the 1 to 3 heterocyclic ring substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, alkylamino of 1 to 4 carbon atoms, dialkylamino of 1 to 4 carbon atoms in each alkyl, carboxy and sulfo; the moiety may be substituted with 1 to 3 substituents selected from the group consisting of hydroxyl, amino, (C 1 -C 4) alkylamino, (C 1 -C 4) dialkylamino, (C 1 -C 4) alkoxy, carboxyl, halogen, and sulfo .

In addition, the ring nitrogen atoms present (except the quaternized nitrogen atom) may be substituted with 1 to 3 substituents independently selected from the group consisting of alkyl of 1 to 4 carbon atoms, alkyl of 1 to 4 carbon atoms substituted with preferably 1 to 3 hydroxy groups, amino groups , (C 1 -C 4) alkylamino, (C 1 -C 4) dialkylamino, (C 1 -C 4) alkoxy, carboxy, halogen or sulfo groups, C 3 -C 6 cycloalkyl, C 3 -C 6 cycloalkylalkyl a (C 1 -C 4) cycloalkyl moiety optionally substituted with 1 to 3 substituents listed above for C 1 -C 4 alkyl, phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group consisting of amino, halogen, hydroxyl, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, and C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino, carboxy and sulfo, C 1 -C 4 phenylalkyl in the alkyl moiety, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and alkyl the moiety may be optionally substituted with 1 to the substituents listed above in connection with C 1-4 alkyl and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms are selected from the group consisting of 1-4 oxygen, sulfur or nitrogen atoms and the alkyl moiety associated with heteroarylalkyl has C 1 -C 6 heteroaryl and heteroarylalkyl optionally substituted in the heterocyclic ring with 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluoromethyl, C 1 -C 4 alkyl, C 1 -C 4 alkoxy carbon atoms, alkylamino having 1 to 5 carbon atoms; C 1-4 alkyl, C 1-4 dialkylamino, carboxyl and sulfo and the alkyl moiety is substituted with 1-3 substituents selected from the group consisting of hydroxyl, amino, C 1-4 alkylamino, C 1-4 dialkylamino in each of alkyl, C1-C18 alkoxy, carboxyl, halogen and sulfo.

is alkyl of 1 to 6 preferably compounds, -CH ₂ ', and wherein

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

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

8

a) R and R together form ch;

or

1

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

CHCHCH _O OH I

Cnebo

OH

I ch ₃ ch-.

CH _O

CH _O , 1 ·

Especially preferred are compounds wherein R₁ is hydrogen and R ^x is

OH

I CH ₃ -chzejména compounds having the absolute configuration 5R, 6S, 8R.

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

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

34 are selected from the group consisting of alkyl of 1 to 6 carbon atoms, OR, -NR R, fluorine, chlorine, bromine, -SO, R 6, -CO-R 6 and -CONR ⁴ R ⁴ , wherein R ³ and R ⁴ in these R ⁴⁴ substituents are jzi as defined in connection with substituent R, or five or six membered heterocyclic ring containing 1-3 heteroatoms selected from the group consisting of oxygen, sulfur, nitrogen or NR ''^'4, wherein R ¹⁴ has the meaning above.

The 5- or 6-membered aromatic quaternized ring or, where appropriate, a carbocyclic, heterocyclic or heteroaromatic ring attached thereto, or both may optionally be substituted on available ring atoms, preferably up to a total of five substituents on the total ring system, the substituents being above in connection with the group

NVE preferred embodiment described above, the preferred compounds are those wherein A is ^- ICH ₂₎ where n is 1 or 2, most preferably those, wherein A is -CH ,, and wherein

8

a) R and R together form a group

CHÍ

1

b) R is hydrogen and R is hydrogen, CH 2 Cl 2 -,

CH,, CH-,

CH, OH: c _? H

CH,

CH,

1

Particularly preferred are compounds wherein R is hydrogen and R is especially those compounds having the absolute configuration of 5R, 6S, 8R.

Another preferred embodiment of the present invention includes compounds of Formula I wherein

NR ⁶ represents a residue selected from the group consisting of

a) the rest of the formula

R ⁵

R ⁷ _R 10 wherein R ⁶ , R ⁷ and R ¹⁸ are independently selected from the group consisting of hydrogen, C 1 -C 4 alkyl, optionally substituted preferably with 1 to 3 hydroxy groups, C 1 -C 4 alkylamino groups, (C 1 -C 4) dialkylamino, C 1 -C 4 alkoxy, amino, sulfo, carboxy or halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine); C 1 -C 4 alkoxy, C 1 -C 4 alkoxy, C 1 -C 4 alkylthio, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino, halogen, (chlorine, bromine) , fluorine or iodine, preferably chlorine, fluorine or bromine), (C 1 -C 4) alkanoylamino, (C 1 -C 4) alkanoyloxy, carboxy, -C-O (C 1 -C 4) alkyl, hydroxyl,

O amidino, guanidino, phenyl, phenyl substituted with one, two or three amino groups, halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyls, trifluoromethyls, (C1-C4) alkyls or alkoxy groups of C 1 -C 4 phenyl, C 1 -C 4 phenylalkyl wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to alkyl of 1 to 4 carbon atoms and heteroaryl and heteroarylalkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms, the alkyl moiety attached to said heteroalkyl moiety has 1 to 6 carbon atoms, or wherein two of the substituents R ®, R ⁷ or R ³⁸ together may be combined to form saturated a fused carbocyclic ring, a fused aromatic carbocyclic ring, a fused non-aromatic, heterocyclic ring or a fused heteroaromatic ring, which fused ring is optionally substituted with 1 or 2 substituents as defined above for R ⁸ , R ⁷ and R ³⁸ ,

(b) formula remnants

R ⁵

R ⁵ '' her or

N

R ^{5 is} optionally substituted on carbon by one to three substituents independently selected from the group consisting of alkyl of 1 to 4 carbon atoms, alkyl of 1 to 4 carbon atoms, substituted preferably with 1 to 3 hydroxy groups, alkylamino groups of 1 to 4 carbon atoms , sulfo, C 1 -C 4 dialkylamino, C 1 -C 4 alkoxy, carboxy, amino or halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C 3 -C 3 cycloalkyl 6 carbon atoms, C 1 -C 4 alkoxy, C 1 -C 4 alkylthio, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino in each alkyl, halogen (chlorine, bromine, fluorine) or iodine, preferably chlorine, fluorine or bromine), (C1-C4) alkanoylamino, (C1-C4) alkanoyloxy, carboxyl, (C1-C4) -alkyl; alkyl, hydroxy, aminidino, guanidino, phenyl, phenyl substituted with one, two or three amino groups, halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyls, trifluoromethyls, alkyls of 1 C 1-4 alkyl or C 1-4 alkoxy, C 1-4 alkyl phenylalkyl, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl, and the alkyl moiety may be optionally substituted with 1 to 3 substituents. above in connection with alkyl of 1 to 4 carbon atoms and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen, or sulfur atoms and the alkyl moiety associated with the heteroarylalkyl moiety Or optionally substituted to form a linked carbocyclic, hete a rocyclic or heteroaromatic ring, optionally substituted with 1 or 2 substituents as defined above?

(c) residues

R ⁵

R ⁵

R ⁵

R ⁵ i \ r tm

R ⁵ .

or

R5

optionally substituted on carbon by one to two substituents independently selected from the group consisting of alkyl of 1 to 4 carbon atoms, alkyl of 1 to 4 carbon atoms, substituted with preferably 1 to 3 hydroxy groups, alkylamino groups of 1 to 4 carbon atoms, sulfo groups of dialkylamino groups (C 1 -C 4) alkyl, (C 1 -C 4) alkoxy, amino, carboxy or halogen (chlorine, bromine, fluorine or iodine, preferably fluorine or bromine chlorine), (C 3 -C 6) cycloalkyl, alkoxy C 1 -C 4 alkylthio, C 1 -C 4 alkylthio, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino in each alkyl, halogen (chlorine, bromine, fluorine or iodine), preferably chloro, fluoro or bromo), (C 1 -C 4) alkanoylamino, (C 1 -C 4) alkanoyloxy, carboxyl, -CO (C 1 -C 4) -alkyl u in alkyl, hydroxy, amidino, guanidino,

Phenyl, phenyl substituted with one, two or three amino groups, halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyls, trifluoromethyls, C1-C4 alkyls or C1-C4 alkoxys carbon, phenylalkyl of 1 to 4 carbon atoms, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above with respect to alkyl of 1 to 4 carbon atoms , and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the aforementioned heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety associated with the heteroarylalkyl moiety has 1 to 6 carbon atoms or is optionally substituted to form a fused carbocyclic, heterocyclic or heteroaromatic ring, optionally substituted 1 or 2 substituents as defined above;

(d) residues of formulas> IM

JM, 5 n

rV

or

optionally substituted on carbon by a substituent independently selected from the group consisting of alkyl of 1 to 4 carbon atoms, alkyl of 1 to 4 carbon atoms, substituted preferably with 1 to 3 hydroxy groups, alkylamino groups of 1 to 4 carbon atoms, sulfo groups, dialkylamino groups of 1 up to 4 carbon atoms in each of the alkyl, C1-C4 alkoxy, amino, carboxy or halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), cycloalkyl of 3 to 6 carbon atoms, alkoxy C 1 -C 4 alkylthio, C 1 -C 4 alkylthio, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino in each alkyl, halogen (chlorine, bromine, fluorine or iodine), preferably chloro, fluoro or bromo), (C 1 -C 4) alkanoylamino, (C 1 -C 4) alkanoyloxy, carboxyl, -GO (C 1 -C 4) -alkyl, hydroxyl yl, amidino, guanidino, phenyl, phenyl substituted with one, two or three amino groups, halogen atoms (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyls, trifluoromethyls, C 1 -C 4 alkyls or alkoxy groups; 1-4 carbon atoms, phenylalkyl having 1 to 4 carbon atoms, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety may be optionally substituted with 1 to 3 subistutents mentioned above with respect to alkyl of 1 up to 4 carbon atoms, and heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the above heterocyclic moieties are selected from the group consisting of 1 to 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety associated with the heteroarylalkyl moiety has 1 to 6 carbon atoms,

(e) remnants of formulas

wherein X is O, S or NR, wherein R is C 1 -C 4 alkyl, C 1 -C 4 alkyl substituted with 1 to 3 hydroxy, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino C 1 -C 4 -alkoxy, C 1 -C 4 -alkoxy, carboxy, halogen or sulfo, C 3 -C 6 -cycloalkyl, C 3 -C 6 -cycloalkylalkyl and C 1 -C 4 -alkyl, optionally substituted with 1 to 3 substituents listed above in connection with alkyl of 1 to 4 carbon atoms, phenyl, phenyl substituted with 1 to 3 substituents independently selected from the group consisting of amino, halogen, hydroxyl, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy (C až-C alky) alkyl (C alky-C alky) alkylamino, (C až-C dial) dialkylamino in each of the alkyls, carboxyl and sulfo, phenylalkyl C 1-4 alkyl in which the phenyl may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to C 1-4 alkyl; and heteroaryl and heteroarylalkyl, wherein the heteroatom or heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl moiety attached to the heteroarylalkyl has 1 to 6 carbon atoms, wherein the heteroaryl and heteroarylalkyl groups are optionally substituted in the heterocyclic ring with 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, halogen, trifluromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, alkylamino of 1 to 4 carbon atoms, dialkylamino of 1 to 4 carbon atoms in each of alkyl, carboxy and sulfo; 3 substituents selected from the group consisting of hydroxyl, amino, alkylamino. C1 -C4 dialkylamino, C1 -C4 dialkylamino, C1 -C4 alkoxy, carboxyl, halogen and sulfo, said heteroaromatic radical being optionally substituted on the carbon atom by one or more substituents independently of one another; selected from the group consisting of alkyl of 1 to 4 carbon atoms, alkyl of 1 to 4 carbon atoms substituted preferably with 1 to 3 hydroxy groups, amino groups, alkylamino groups of 1 to 4 carbon atoms, dialkylamino groups of 1 to 4 carbon atoms in each alkyl, (C 1 -C 4) alkoxy, sulfo, carboxy or halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), C 3 -C 6 cycloalkyl, C 1 -C 4 alkoxy, C 1 -C 4 alkylthio C 1 -C 4, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino, in each residue, halogen (chlorine) (bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), (C 1 -C 4) alkanoylamino, (C 1 -C 4) alkanoyloxy, carboxy, -C (C 1 -C 4) alkyl, hydroxyl, amidino , guanidino, phenyl, phenyl substituted with one, two or three substituents such as amino, halogen (chlorine, bromine, fluorine or iodine, preferably chlorine, fluorine or bromine), hydroxyl, trifluoromethyl, (C 1 -C 4) alkyl, or (C 1 -C 4) alkoxy, (C 1 -C 4) phenylalkyl, wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above with respect to alkyl having 1 to 4 carbon atoms, heteroaryl or heteroarylalkyl, wherein the heteroatom or heteroatoms in the above-mentioned heterocyclic moieties are selected from the group consisting of 1 to 4 carbon atoms; 4 oxygen, nitrogen or sulfur atoms and the alkyl moiety attached to said heteroarylalkyl moiety has 1 to 6 carbon atoms or is optionally substituted to form a fused carbocyclic, heterocyclic or heteroaromatic ring, optionally substituted with 1 or 2 substituents as defined above;

(f) formula remnants

R5

\ Φ

N —X - (ΓΊΙ © c hk ^ N-R “

NR ϋ π — NR ⁵ i | j_ ^Ν ? Χ ^

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

(g) formula remnants

n_n-r II I c '

Nx = NR ⁵ "$ _r ~ NR 5_n-

II I

N ^ N or n-n-R

wherein R is C 1 -C 4 alkyl, C 1 -C 4 alkyl substituted with 1 to 3 hydroxy, amino, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino, C 1 -C 4 alkoxy 4 carbon atoms, carboxy, halogen or sulfo groups, cycloalkyl of 3 to 6 carbon atoms, cycloalkylalkyl of 3 to 6 carbon atoms in the cycloalkyl moiety and 1 to 4 carbon atoms in the alkyl moiety, optionally substituted with 1 to 3 substituents listed above in connection with C 1-4 alkyl, phenyl, phenyl substituted with 1-3 substituents independently selected from the group consisting of amino, halogen, hydroxyl, trifluoromethyl, alkyl of 1 to 4 carbon atoms, alkoxy of 1 to 4 carbon atoms, alkylamino of (C 1 -C 4), (C 1 -C 4) dialkylamino, carboxy and sulfo, (C 1 -C 4) phenylalkyl; an alkyl moiety wherein the phenyl moiety may be optionally substituted with 1 to 3 substituents listed above in relation to phenyl and the alkyl moiety may be optionally substituted with 1 to 3 substituents listed above with respect to alkyl of 1 to 4 carbon atoms, heteroaryl and heteroarylalkyl, wherein the heteroatom and the heteroatoms are selected from the group consisting of 1 to 4 oxygen, sulfur or nitrogen atoms and the alkyl moiety associated with the heteroarylalkyl has 1 to 6 carbon atoms, said heteroaryl and heteroarylalkyl groups being optionally substituted in the heterocyclic ring with 1 to 3 substituents independently selected from the group including hydroxyl, amino, halogen, trifluoromethyl, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, C 1 -C 4 alkylamino, C 1 -C 4 dialkylamino in each alkyl, carboxy, and sulfo group; and the alkyl moiety is substituted with 1 to 3 substituents independently selected from the group consisting of hydroxyl, amino, (C 1 -C 4) alkylamino, (C 1 -C 4) dialkylamino, (C 1 -C 4) alkoxy, carboxyl, halogen and sulfo.

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

Among the preferred embodiments of the present invention described above, those wherein A is ^CH 2 * n ^and n ° C are preferred, and more preferred are those wherein A is Cl 2 " ^and wherein

8

(a) R and R together form ^HCH 2 or

1

b) R is hydrogen and R is hydrogen, CH 2 Cl 2 -

Target., 3 ^CH 3- OH OH C or CH ₃ CH-. ^CH r ^ ^ch 3 Particularly preferred are compounds. where R ⁸ is hydrogen and is

OH (

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

A particularly preferred embodiment of the present invention includes compounds of formula I wherein it represents the remainder of the formula

wherein r ⁶ , r ⁷ and are independently selected from hydrogen, C 1 -C 4 alkyl, C 1 -C 4 alkoxy, carboxyl and carbamoyl, and R ³ is as defined above and is preferably C 1 -C 4 alkyl; up to 6 carbon atoms, preferably methyl.

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

a) R and R together form

BOY

C =

CH or

1

b) R c is hydrogen and R c is hydrogen, CH 2 CH 2,

CHCH-, ^CH 3 of CH 2 CH 2 CHCHCH 1

1

Particularly preferred are compounds wherein R is hydrogen and R is

Especially those compounds having the absolute configuration of 5R, 6S, 8R.

Another preferred embodiment includes compounds of Formula I wherein

N ® -R ⁵ represents the remainder of the formula

and)

wherein R ³ is C 1 -C 4 alkyl, preferably methyl, and R 3 is hydrogen or C 1 -C 4 alkyl,

b) _n 5

Wherein R ⁵ is C 1 -C 4 alkyl, preferably methyl, and R ⁵ and R ⁷ are hydrogen or C 1 -C 4 alkyl, ^C 1 -C ₅

RN NR ⁵ wherein R is alkyl of 1 to 4 carbon atoms, preferably methyl and R is alkyl having 1 to 4 carbon atoms or phenylalkyl having 1 to 4 carbon atoms,

(d)

ŇTŇ ^ rG

R 0 wherein R ⁵ is C 1 -C 4 alkyl, preferably methyl, and R ⁵ is C 1 -C 4 alkyl.

preferably methyl,

E)

RS '©

-C)

Wherein R is alkyl of 1 to 4 carbon atoms, preferably methyl and R is alkyl of 1 to 4 carbon atoms, preferably methyl or

f) wherein R ⁵ is C 1 -C 4 alkyl, preferably methyl.

From the above-described embodiment of the present invention, preferred compounds are those wherein A is ^{- '-CH} 2> N ^{-' N} 1 ^e 3 ° ^NEB 2 ^»3 preferably those compounds wherein A is -Cl ^ -, and wherein

8

a) R and R together form

or

1

b) R is hydrogen and R is hydrogen, CH ₃ CH ₂ -,

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

especially those compounds having the absolute configuration of 5R, 6S, 8R.

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

4- nr ⁵ represents the rest of the formula

(b)

(d)

F)

(h)

(j)

1)

(n)

CH

CH, _CH3 / in

N ^ N ^ CH ₃

Of this embodiment described above, those compounds wherein A is - ( ^CH 2) n ^- are 1 or 2, more preferred are those compounds wherein A is - (C 1-6) - and wherein

8

(a) R and R together form boy ₂

or

b) R 0 is hydrogen and R ¹ is hydrogen, CH ₃ CH ₂ -,

CH.

CH.

Particularly preferred are those wherein R ⁸ is or a hydrogen atom and R 8 is

Especially those compounds having the absolute configuration of 5R, 6S, 8R.

Particularly preferred compounds of the present invention are compounds of formula

wherein R is a hydrogen atom, an anionic charge, or a conventional, easily cleavable carboxylic acid protecting group, wherein when R is a hydrogen atom or a protecting group it may also be present in the form of an ion and wherein

-S-A

NR ⁵ is

a) _CH ₂ - / - CH,

b) ₂ -5ΟΗ ΟΗ ₂ -ΑΛ | -ΟΗ3

CH,

d) - SCH;

f) -sch ₂ - <(n-ch ₂ ch ₂ ch ₃ where

3.12

@ 1 H NMR ( ^D @ 20) shows characteristic peaks at delta: 1.23 (3H, d, J = 6.4 Hz), (2H, q, J = 1.4, 8.9 Hz), 3.39 ( 1H, q, J = 2.7, 6.0Hz), 4.07-4.68 (10H, m), 8.19 (1H, s);

q) ^- SCH '

CH,

CH, where

3.15

NMR (D ₂ O) spectrum shows characteristic peaks at delta 1.23 (3H, d, J = 6.4 Hz), (2H, q, J = 3.7, 9.0 Hz), 3.37 ( 1H, q, J = 2.6, 6.0 Hz), 3.95-4.65 (10h, m), 8.62 (1H, s);

(r)

SCH _S / ^{N CH3}

with)

CH,

t) φ

-sch ₂ - / \

at)

COO .Θch ₂ i® sch ₂ -ZV ¹ n-ch ₃

CH, n-ch ₂ coo ' ^SCH Z

N-N /

ch ₃ or

CH

₃

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

Of the above-described embodiment of the present invention, those compounds wherein A 18 is "CH ₂ ) _n , where n is 1 or 2, are preferred, wherein A is -CH ₂ - and wherein a) R and R together form a boy" ₂ or

b) R ⁸ is

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

CH_

CH3OH 3-. I? H-,

OH ch ₃ Íh-.

Particularly preferred are compounds wherein R is hydrogen

OH l

_CH3 CHzejména those compounds

The carbapenem derivatives of formula II have an absolute 5R configuration, of formula I being prepared

6S, 8R.

from raw materials of general use

R ⁸ H

H.-šor '15

I .S A

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

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

d ') has a balancing ion X' (for example derived from the alkylating agent used), which at this stage or at a later stage, for example after the cleavage step, can be replaced by another ion, for example in a conventional manner, for a more pharmaceutically suitable ion. Alternatively, the ion may be removed during the reaction by leaving the protecting group.

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

'ovv, _at ,

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

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

Similarly, other conventional carboxyl protecting groups can be removed by methods commonly known in the literature. Finally, as mentioned above, compounds of formula I wherein R is a physiologically hydrolysable ester such as acetoxymethyl, phthalidyl, indanyl, pivaloyloxymethyl, methoxymethyl and the like can be applied directly to the host without deprotection, as these esters are hydrolyzable in vivo under physiological conditions.

5 15

It is understood that where an R, R, R, or R substituent or heterocyclic ring is attached to a substituent A containing a functional group that may interfere with the reaction being contemplated, that group may be protected by a conventional protecting group and then removed. regenerates the desired functional group.

Suitable protecting groups and procedures for introducing and removing these groups are well known in the literature.

In a variant of the above process, the carboxyl protecting group of intermediate II can be removed prior to the quaternization step. Thus, the carboxyl protecting group is removed as described above to give the corresponding free carboxylic acid and the free acid is then quaternized with an alkylating agent R-X 'to give the desired quaternized product of Formula I.

When the unprotected intermediate IIIa is quaternized, water or a non-reactive organic solvent or a mixture thereof can be used as the solvent. Examples of suitable solvents are water, organic solvents such as chloroform, methylene chloride, tetrahydrofuran, dioxane, acetone, dimethylsulfloxide and dimethylformamide and water-organic solvent mixtures such as water-acetone or water-dimethylformamide.

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

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

It has been found that this greatly facilitates preferential quaternization at the ring nitrogen atom.

The above-described variant is particularly useful when the carboxyl protecting group is more easily removable from the non-quaternized intermediate XI than from the quaternized intermediate I. For example, in the preparation of a product of the formula

R) (R) from an intermediate of formula

removal of the allyl protecting group before quaternization results in better yields of the desired end product.

The starting compounds of formula (J) used in the process of the present invention can be prepared from compounds of formula (III)

(III) wherein r 1, R 2 and R 2 are as defined above, and wherein R ² represents a conventional readily cleavable carboxyl protecting group. The compounds of formula III are disclosed, for example, in European Published Patent Application No. 38,869 (Compound 7) and European Published Patent Application No. 54,917 and can be prepared by the general methods described above.

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

*

5 »»

L = normal leaving group

A variation of the procedure described above is apparent from the following reaction scheme

R® o1 --- 1 - = _d 15

HS-A

COOR ²

R®

R ¹⁵

S A

COOR ² deprotection

 (III

When operating as described above, the starting material of formula XII is reacted in an inert organic solvent such as methylene chloride, acetonitrile, or dimethylformamide with an approximately equimolar amount of reagent R ° -L, such as p-toluenesulfonic anhydride, p-nitrobenzenesulfonic anhydride. acids, 2,4,6-triisopropylbenzenesulfonic anhydride, methanesulfonic anhydride, trifluoromethanesulfonic anhydride, diphenylchlorophosphate, toluenesulfonyl chloride, p-bromobenzenesulfonyl chloride and the like, wherein L is a corresponding leaving group such as toluenesulfonyloxy group, p-nitrobenzenesulfonyloxy group, other p-nitrobenzenesulfonyloxy group and other which are cleavable by conventional methods known in the literature.

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

The leaving group L of intermediate IV may also be a halogen atom, in which case this group is cleaved by reacting intermediate III with a halogenating agent such as O 2 PCl ₃ and (O) 2 PB 3, oxalyl chloride and the like in a solvent such as dichloromethane, acetonitrile, tetrahydrofuran and the like, in the presence of a base such as diisopropylethylamine, triethylamine, 4-dimethylaminopyridine and the like.

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

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

wherein A is an unbranched or branched alkylene chain of 1 to 6 carbon atoms and represents a mono-, bi- or polycyclic aromatic heterocyclic radical containing a nitrogen -izable quaternary ring, which ring is attached to A through the carbon *. in an inert organic solvent such as dioxane, dimethylformamide, dimethylsulfoxide or acetonitrile and in the presence of a base such as diisopropylethylamine, triethylamine, sodium bicarbonate, potassium carbonate or 4-dimethylaminopyridine.

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

While the above process is suitable for the preparation of the compounds of the present invention, a colleague of Pierre Dextraze has found a new process for the preparation of the compounds of Formula I. This alternative process, which is disclosed and claimed in the parallel US patent application filed the same day as this application, .

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

(IV) wherein R ⁸ and R ⁸ are as defined above, a conventional readily removable carboxyl protecting group is common and L is a common leaving group such as toluenesulfonyloxy, β-nitrobenzene; VII

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

R ° H

NR ³

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

O COOR ²

d ') wherein R ¹ , R ⁸ , R 1', A, R ¹ ', 2' and X are as defined above and optionally the carboxyl protecting group R is removed to give the corresponding deprotected compound of formula I or a pharmaceutically acceptable salt thereof .

An alternative procedure utilizes the intermediate formula

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

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

The intermediates of formula IV are generally formed in situ by reaction of an intermediate of formula III

R ⁸ H _r 15

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

The carbapenem intermediate IV is reacted with a quaternary aminthiolein of formula

VII

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

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

1,8-diazabicyclo (5.4.0) undec-7-ene, 1,5-diazabicyclo (4,3,0) non-5-ene and C 1 -C 4 trialkylamines in each of the alkyls, such as triethylamine, tributylamine or tripropylamine.

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

Carbapenem products prepared by the reaction of a quaternary aminthiol VII with an intermediate of formula IV are associated with an anion (for example (C CH HO) ₂ ?? ₂ Θ, Cl θ or an anion associated with a quaternary thiol), which at this stage may be replaced by another anion , for example, one that is pharmaceutically aqueous. This replacement is carried out in a conventional manner. Alternatively, the anion can be removed during the next deprotection step.

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

To form the desired carbapenem product, the carboxyl protecting group R ₂ in compound I 'can optionally be removed by conventional methods such as solvolysis, chemical reduction or hydrogenation. If a protecting group such as p-nitrobenzyl, benzyl, benzhydryl or 2-naphthylmethyl is used which can be removed by catalytic hydrogenation, then. intermediate I * in a suitable solvent such as dioxane-water-ethanol, tetrahydrofuran-diethyl ether-buffer, tetrahydrofuran-aqueous potassium hydrogen phosphate-isopropanol, or the like. They can be reacted under a hydrogen pressure of from 0.1 to 0.4 MPa in the presence of a hydrogenation catalyst such as palladium on carbon, palladium hydroxide, platinum oxide and the like at a temperature of from 0 to 50 ° C for 0.24 to 4 hours.

If there is a group such as o-nitrobenzil, photolysis can also be used to remove the protecting group. Protecting groups such as 2,2,2-trichloroethyl can be removed by slight reduction with zinc.

The allyl protecting group may 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 may be removed by methods commonly known in the art. Finally, as noted above, the compounds of formula I wherein R is a physiologically hydrolyzable ester such as acetoxymethyl 'phthalide' .- ¹ iudanyl, pivaloyloxymethyl, methoxymethyl, etc.. Aplikovič may directly hostiie.lj OEZ deprotection monoperoxycarbonate or these esters, they are hydrolysable in vivo under physiological conditions.

The thiol intermediates of formula VII can be prepared, for example, from the corresponding thioacetate compounds of the formula

wherein A is as defined above and

represents a mono-, bi- or polycyclic aromatic heterocyclic radical containing a quaternary ring-atomizable nitrogen atom, which ring is attached to A via a ring carbon atom. dioxane, benzene, xylene, toluene or mixtures thereof with a suitable alkylating agent of the general formula

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

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

By suitably selecting solvents, the reaction in intermediate III to the final product I can be carried out without isolation of the various intermediates, i.e. by the so-called single flask process.

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

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

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

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

It will be understood that certain products within the scope of formula (I) may be formed in the form of optical isomers as well as in the form of epimeric mixtures. All optical isomers and epimeric mixtures are included within the scope of the present invention.

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

A compound of formula 1 wherein R is hydrogen or an anionic charge, or a pharmaceutically acceptable 2 thereof.

a suitable salt may also be converted by a conventional method to the corresponding compound wherein R is a physiologically hydrolyzable ester group or a compound of formula I wherein R is a common carboxyl protecting group 2 may be converted to the corresponding compound wherein R is a hydrogen atom, an anionic charge, or a physiologically hydrolyzable ester group, or a pharmaceutically acceptable salt thereof.

Novel derivatives of carbapenem of formula I, wherein R ² is hydrogen, an anionic charge or a physiologically hydrolysovateiná carboxyl protecting group, or their pharmaceutically acceptable salts are potent antibiotics for various gram-positive and gram-negative bacteria and can be used for example as additives to growth-enhancing feedstuffs, as food preservatives, as bactericides for industrial applications, for example, water-soluble dyes and orbital waters in paper mills, where they inhibit the growth of harmful bacteria and as disinfectants for decomposition and inhibition of harmful bacteria growth in medical and dental instruments .

However, they are particularly useful in the treatment of infectious diseases in humans and other animals caused by Gram-positive or Gram-negative bacteria.

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

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

Injectable compositions, a preferred mode of administration, may be prepared in unit dosage form in ampoules or in multi-dose containers, and may contain adjuvants such as suspending, stabilizing and dispersing agents. The compositions may be in a form directly applicable for application or in a powder form for reconstitution at a suitable time by the addition of a suitable carrier, such as sterile water.

The dose to be administered depends in particular on the compound used, the particular formulation of the composition, the mode of administration, the type and condition of the patient and the target site, and the organism to be treated. The choice of a particular preferred dosage and route of administration is then left to the skill of the art. In general, however, the compounds are administered parenterally or orally to a mammal in an amount of from about 5 to 200 mg / kg / day.

Administration is generally carried out in divided doses, i.e. three to four times a day.

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

In vitro activity

Samples of the carbapenem compounds prepared in Examples 1 and 2, when dissolved in water and diluted with nutrient medium, exhibited the following minimum inhibitory concentrations (MIC) in µg / ml on said microorganisms. The assay was determined by incubating overnight at 37 ° C- by dilution in a test tube. N-formimidoylthiermamycin was used as a reference compound.

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

MIC (^ g / ml)

New compound N-formimidoylthienamycin

with. pneumoniae A-9585 0.25 0.004 with. Pyogenes A-9604 0.06 0.001 with. Aureus A-9537 0.13 0.004 with. aureus + 50% serum A-9537 0.03 0.016 with. aureus (Pen-res.) 1-9606 0.06 0.008 with. aureus (Meth res.) A-15097 4 0.5 with. faecalis A20688 0.5 0.5 E. coli (10 ⁴ dil.) A15119 0.06 0.016 E. coli (10 ^-3) A15119 - 0.03 E. coli (ΙΟ ^-2 ) A15119 - 0,06 ' E. coli (10 ~ ⁴⁾ A20341-1 0.03 0.03 E. coli (10 ^-3 ) A20 341-1 - 0.03 E. coli, into ^'2) A20341-1 - 0.13 TO. pneumoniae A-9664 0.25 0.13 TO. pneumoniae A20468 0.25 0.06

continued table

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

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

MIC (µg / ml)

New compound N-formimidoylthienamycin

with. pneumoniae A-9585 0.001 0,002 with. pyogenes A-9604 0.001 0,002 with. Aureus A-9537 0.004 0.004 with. aureus A-9537 + 50% serum 0.016 0.016 with. aureus A-9 6 0 6 (Pen-re) 0.008 0.008 with. aureus (Meth-res.) A15097 8 4 with. faecalis A20688 0.25 0.5 E. coli A15119 0.016 0.016 E. coli A2034L-1 0.016 0.03 TO. pneumoniae A-9664 0.06 0.06 P. mirabilis A-9900 0.03 0.06 P. vulgaris A21559 0.016 0.03 P. morganii A15153 0.06 0.13 P. rettgeri A22424 0.13 0.13 with. marcescens A20019 0.03 0.03 E. cloacae A9659 0.13 0.06 E. cloacae A9656 0.25 0.06 P. aeruginosa A9843A 8 1 P. aeruginosa A21213 2 0.25 In vivo activity In vivo therapeutic effect the compound of Example 1 and N, N-formimidoylthienamycin

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

Protective effect in intramuscular treatment of infected mice

Organism Infection (number organisms) PDj-0 (treatment (mg / kg) Compound according to Example 1 N-formimidoyl- thienamycin P. , mirabilis A-9900 3.6 χ 10 ⁶ 3.3 3 ^X / 15 ^x P. . aeruginosa A-9843a 5.5 χ 10 ⁴ 0.3 0.5 ^x P. . aeruginosa A-20481 5.4 χ 10 ⁴ 0.63 0.4 ^X P, , aeruginosa A-20599 1.4 χ 10 ⁵ 0.7 0.18 ^x with. aureus A-9606 6.6 χ 10 ~ ⁸ 0.09 0,07 ^x with. faecalis A-20688 2.3 χ 10 ⁸ 3.3 2.8 ^X E. coli A-15119 6.2 x 10 ⁶ 0.6 2,5 ^X TO. . pneumoniae A-9964 5.1 x 10® 2.5 2,2 ^X

historical data sequence of treatments: With the exception of E. coli A15119 and K. pneumoniae A9964, mice are treated i.m. drug 0 and 2 hours after infection. For E. coli and K. pneumoniae treatment was 1 and 3.5 hours after infection. 5 mice per dose were used for each test.

Toxicity

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

Toxicity for intracranial administration to mice

Compound

Highest dose (mg / kg) x _T No clinical ^Βυ 50 signs of toxicity (mg / kg) Compound of Example 1

N-formimidoylthienamycin

5

2 x 5 ^x average 25 / mouse / compound

Blood levels of mice after intramuscular administration

Blood levels and half-life of the compound of Example / 0 mg / kg mice are shown in the following table:

after intramuscular administration

Compound 10 20 May Level v 30 45 blood 60 (µg / ml) 90 ^x tl / min (min) ^XX AUC (µg / h) the compound of Example 1 14 10.8 6.8 2.6 0.8 Zl · 0.6 10 6.3 N-formimidoylthienamycin 12.6 9.9 7.3 2.6 0.7 <0.3 9 6

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

Excretion of urine

Urine / compound excretion in Example 1 following intramuscular administration (20 mg / kg) to mice is seen from n / sec; . : * · Ϊ> '·.

Urinary excretion following intramuscular administration to 20 mg / kg to mice

Compound Percentage of dose excluded 0-3 3-6 hours after 6-24 application 0-24 the compound of Example 1 26.1 0.5 0.1 26.7 + 6.7 N-formimidoylthienamycin 12.1 0.1 <0.1 12.2 + 3.6

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

In vitro activity

Samples of the carbapenem compounds listed below (identified by example numbers) for dissolution in water and dilution with nutrient medium exhibited the following minimum inhibitory concentrations (MIC) in yug / ml for the indicated microorganisms. Test performed by incubation overnight at 37 ° C by dilution in a tube. In the following tables, N-formimidoylthienamycin is used as reference compound.

MIC (µg / ml)

Ex 3 4 Ex 5 Ex 6 Ex 7 MK 0787 with. , pneumoniae 1-9585 0,002 0,002 0.004 0.004 0.004 0,002 with. pyogenes A-9604 0,002 0.001 0.004 0.004 0.004 0,002 with. , faecalis A20688 0.5 0.5 0.25 0.5 0.13 0.5 with. . aureus A-9537 0.016 0.008 0.004 0.03 0.008 0.004 5. . aureus + 50% serum 1-9537 0.016 0.03 0.016 0.06 0.03 0.016 with. aureus A-9606 Pen-res 0.016 0.016 0.008 0.03 0.016 0.008 WITH. aureus A15097 4 4 8 4 2 4

Meth-res

E. coli A15119 0.03 0.016 0.016 0.06 0.004 0.016 E. coli A20341-1 0.016 0.016 0.016 0.06 0.004 0.03 TO. pneumoniae A-9664 0.06 0.03 0.03 0.13 0.016 0.06 TO. pneumoniae 120468 0.06 0.03 0.06 0.25 0.016 0.13 E. clocae A-9659 0.13 0.03 0.06 0.25 0.016 0.06 E. clocae A-9656 0.13 0.06 0.13 0.25 0.016 0.06 P. mirabilis A-9900 0.13 0.06 0.03 0,025 0.016 0.06 P. vulgaris A-21559 0.016 0.016 0.016 0.06 0.008 0.03 M. morganii A15153 0.13 0.016 0.06 0.13 0.016 0.13 P. rettgeri A22424 0.25 0.13 0.13 0.25 0.06 0.13 WITH. marcescens A20019 0.016 0.03 0.06 0.13 0.008 0.03 P. aeruginosa A-9843a 2 2 4 8 1 0.25 P. aeruginosa A21213 0.5 0.13 2 1 0.25 0.25 H. influenzae A-9833 2 2 4 > 32 > 32 16 H. influenzae A21518 2 2 4 732 > 32 16 B. fragilis A22862 0.25 0.06 0.03 0.03 0.016 0.06 B. fragilis A22696 0.5 0.5 0.25 0.25 0.25 0.13

^x N-formimidoylthienamycin

Organism Ex 8 MIC (µg / ml) Ex 9 eg 10 Ex 11 MK 0787 WITH. pneumoniae A-9585 0,002 0.001 0.001 0,002 0,002 WITH. pyogenes A-9604 0,002 0.001 0.001 0,002 0,0Q2 WITH. faecalis A20688 0.5 0.13 0.25 0.5 0.25 WITH. aureus A-9537 0.008 0.004 0.008 0.004 0? 002 WITH. aureus A-9537 0.03 0.008 0.06 0.016 0.016

(50% serum)

WITH. aureus A-9606 0.03 0.008 0.008 0.016 0.008 Pen-res with. aureus A15097 - - - Meth-res E. coli A15119 0.016 0.008 0.016 0.016 0.016 E. coli A20341-1 0.03 0.004 0.008 0.03 0.016 TO. pneumoniae A-9664 0.03 0.016 0.06 0.03 0.03 TO. pneumoniae A20468 0.13 0.03 0.13 0.13 0.13 E. clocae A-9659 0.13 0.03 0.013 0.06 0.13 E. clocae A-9656 0.06 0.03 0.13 0.13 0.06 P. mirabilis A-9900 0.13 0.016 0.13 0.03 0.03 P. vulgaris A21559 0.03 0.008 0.016 0.03 0.016 M. morganii A15153 0.13 0.03 0.13 0.06 0.06 P. rettgeri A22424 0.13 0.06 0.13 0.13 0.13 WITH. marcescens A20019 0.06 • 0.016 0.06 0.06 0.03 P. aeruginosa A-9843A 1 2 32 0.5 1 P. aeruginosa A21213 0.25 0.13 2 ' 0.13 0.13 ^X N- -formimidoylthienamycin

Organism pr. 12 MIC (µg / ml) Ex 13 Ex 14 MK 0787 ^X WITH. , pneumoniae A-9585 0,002 0.0005 0.0005 0,002 WITH. pyogenes A-9604 0.004 0.0005 0.0005 0,002 WITH. faecalis A20688 0.5 0.13 0.13 0.25 WITH. . aureus A-9537 0.008 0.008 0.008 0,002 with. aureus A-9537 0.016 0.016 0.03 0.008 + 50% serum with. aureus A-9606 0.03 0.008 0.016 0.008 Pen-res with. aureus A15097 - - - - Meth-res E. coli A15119 0.016 0.004 0.008 0.016 E. coli A20341-1 0.008 0.008 0.008 0.016 TO. pneumoniae A-9664 0.03 0.03 0.03 0.03 TO. pneumoniae A20468 0.06 0.13 0.03 0.06 E. cloacae A-9659 0.06 0.06 0.03 0.06 E. cloacae A-9656 0.03 0.03 0.03 0.06 P. mirabilis A-9900 0.03 0.016 0.016 0.016 P. vulgaris A21559 0.016 0.008 0.016 0.016 M. morganii A15153 0.06 0.016 0.03 0.06 P. rettgeri A22424 0.13 0.25 0.06 0.13 WITH. marcescens A20019 0.03 0.016 0.016 0.03 P. aeruginosa A-9843A 16 32 8 1 P. aeruginosa A21213 2 2 0.5 0.13

^X N-formimidoylthienamycin

In vivo activity

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

Values of ΡΟ ₅ θ (dose in MG / kg) that provide 50% protection of infected mice are shown.

Protective effect in intramuscular treatment of infected mice

PDjg / treatment (mg / kg)

Ex 6 Ex 8 Ex 9 Ex 12 Ex 14 MK 0787 with. aureus A9606 - 0.21 - 0.89 0.07 0.07 E. coli A15119 - 0.86 1,2 - - 3 TO. pneumoniae A9664 - 1,8 1,8 - - 3 P. mirabilis A9900 - 1.4 7.1 - - 9 P, aeruginosa A9843A 0.4 0.19 0.19 1,8 0.45 1 P, aeruginosa A24081 - 0.33 0.19 - - 0.4

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

Blood levels of mice after intramuscular administration

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

Compound C max Qig / ml) * ^T l / 2 (min) ^XX AUC (jog.h / ml) example 1 14 10 6.3 Example 2 13.9 9 5.3 Example 3 14.5 10 • 6.9 Example 4 15.5. 11 7.7 Example 5 - - Example 6 17.7 9 8.5 example 7 19.2 11 11.8 Example 8 18.8 11 10.5 Example 9 16.7 12 Ó, 5

continued table

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

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

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

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

Example 1

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

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

%

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

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

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

NMR (DMSO-d6) δ:

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

4.1 (2H, m);

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

J = 5.5 Hz)

IC (KBr) gamma max: 3400,

790,

695 and 1600 cm

Anal. Calcd for ^C 22 ^H 21 ^N 3 ° 6 ^S

Found: C, 57.74; H, 4.56; N, 9.58; N, 7.04; N, 7.21.

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

NMR (DMSO-d 6) δ: 1.15 (3H, d, J = 6.0 Hz), 3.2 to 3.4 (3H, m), 3.7 to 4.1 (2H, m),

4.25 (3H, s), 4.30 (2H, s), 5.25 (1H, d, J = 14.0 Hz), 5.50 (1H, d, J = 14.0 Hz), 7, 70 (2H, d, J = 9.0Hz), 8.10 (2H, d, J = 7.0Hz)

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

IR (KBr) gamma: 3400, 1770, 1690 and 1640 cm ^-1 .

max ·

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

To a solution of 779 mg (1.27 mmol) of 3 in tetrahydrofuran-ether (80 mL-80 mL-100 mL) was added 140 mg (1.4 mmol) of dibasic potassium phosphate bicarbonate (0.7 mmol).

water and diethyl potassium and 125 mg

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

This crude material was purified on a C18 g BONDAPAK reverse phase column (30 g) (Waters Associates) and eluted with water at 50 psi. Each fraction (20 mL) was separated by high pressure liquid chromatography, and the ultraviolet absorption fractions at lambda _max = 300 nm were combined and lyophilized to afford 135 mg (32% yield) of compound 4 as a pale yellow solid.

NMF t (D ₂ O) delta: 1.25 (3H, d, J = 6.0) and 2.5 Hz), 3.9-4 d, J = 6.0 Hz), 8.72 IR (KBr) ^gamma max : 3 400, 1755, 1640 UV lambda max ^(H 2 ') 0): 296 nm (epsilon = ř í k lad 2

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

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

OH

S · ©

N- CH

OH

CO ₂ pNB

CO ₂ PNB

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

IR (KBr) 1765, 1690 cm

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

B.

CHq

CO ₂ pNB (Η) γ

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

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

The aqueous layer was lyophilized to give a brownish solid which was purified twice by chromatography on 12 g C ^ gBONDAPAK reverse phase column (Waters Associates) (H ₂ O) to give 55 mg of the fifth

IR (KBr): 1750, 1640 cm @ ³ .

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

OH

ch ₃

Example 3

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

CH ₃ I

OH

Pd / C, H ₂

CH,

COjpNB

OH

CH,

WHAT,

1 '

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

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

NMR (DMSO-d6) δ: 1.25 (3H, d, J = 5.8 Hz), 3.6-4.3 (5H, nI), 4.20 (3H, s), 4.25 ( 2H, s), 5.25 (1H, d, J = 7.2 Hz), 5.40 (2H, ABq, J = 12.16 Hz) and 7.6-9.2 (9H, m).

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

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

To a solution of 933 mg (1.6 mmol) of compound 20 in 90 mL of tetrahydrofuran and 90 mL of ether was added 200 mg of KHCO3 and 249 mg of &lt; RTI ID = 0.0 &gt; HPO2 &lt; / RTI &gt; in 90 mL of water.

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

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

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

Analysis for ^C ^ gN IgH ₂ O ^ S ^ .2H ₂ O calculated 51.87% C 5.44% H 7.56% N Found 51.95% C, 5.66% H, 7.56 % N.

Example 4

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

OH

CO ₂ pNB

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

To a solution of compound 22 (650 mg, 1.39 mmol) in acetone (100 mL) was added methyl iodide (4 mL). The reaction mixture was stirred at room temperature for 3 days. The precipitate was filtered off and washed with acetone (10 mL) to give 500 mg (60% yield) of quaternized pyridine 26 as a light yellow solid.

NMR (DMSO-D 6) δ 1.26 (3H, d, J = 7.0 Hz), 3.9-4.2 (2H, m), 4.4 (3H, s), 4.78 (2H s), 5.2 (1H, d, J = 3.8 Hz), 5.50 (2H, ABq, J = 14 Hz) and 7.8-9.4 (8H, m).

IR (KBr) gamma 3400, 1765 and 1690 cm @ -1 max

Analysis for ^C 23 ^H 24 ^N 6 ^S 3 ° l ^'i' l calculated 46.24% C, 4.05% H, 7.03% N, 5.37%; Found 45.62% C 4, % H,% N, 5.30%.

To a solution of compound 23 (1.0 g, 1.167 mmol) in tetrahydrofuran (90 mL) and ether (90 mL) was added KHCO 3 (215 mg, 2.15 mmol) and K 4 HPQ 4 (374 g, 2.1 mmol) in water (90 mL). 1.0 g of 10% palladium on carbon is added. The mixture is hydrogenated at 310 kPa for 45 minutes on a Paar shaker.

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

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

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

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

TC (KBr) 9 ^ama max ^: 3,400, 1,755 and 1,590 cm.

UV λmax (H-O): 292 nm (epsilon = 8092). max z *

Analysis for ^C i6 ^H 28 ^N 4 ^S 2 ° L ^'2H 2 ° Calculated 51.87% C 5.44% H 7.56% N Found 51.37% C, 5.69% H, 7 37% N.

Example5 &

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

CO ₂ pNB

- 1- (R) -hydroxyethyl-3- (N-methylpyridin-2-ylethanthio) -6alpha-hept-2-ene-2-carboxylate

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

NMR (DMSO-d 6) δ: 1.20 (3H, d, J = 5.6 Hz), 3.2-4.3 (9H, m), 4.20 (3H, s), 5.26 and 5.55 (1H each, ABq, J = 15 Hz) and 7.8-9.2 (8H, m).

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

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

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

10% acetonitrile in water at 55 kPa.

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

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

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

750, 1590 cm ¹ .

= 8,108).

Preparation of 3- (1-propylpyridin-4-ylmethanthio-6α) - [11- (E) hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

acetone

Nal

- (1-Allylpyridin-4-ylmethanthio) -6alpha- [1- (R) -hydroxyethyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-3-2-carboxylate

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

This material is suspended in 120 ml of acetonitrile, p. jt is sc and ee ^{, 7} g to give 1.0 g (87% yield) of the title compound as a yellow solid. and·'.·.:;.·.

NMR (CD CH) δ 1.20 (3H, d, J = 6.2 Hz), 3.0 to 3.4 (4H, m), 4.0 to 4.4 (4H, m), δ 1 to 5.6 (4H, m) and 7.4 to 7.9 (8H, m).

IR (KBr) gamma: 3400, 1770 and 1690 cm

Analysis for ^C 25 ^RI 26 ^N 6 ^S 3 ° l 'l ^I * calculated 48.16% C, 4.21% H, 6.74% N, 5.15% G, Found 48.55% C 4, H, 46%; N, 6.69; S, 5.15.

24719

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

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

The reaction mixture was filtered through celite and the catalyst washed with water (2 x 10 mL) and lyophilized to give a yellow powder which was purified on a C _lg BONDAPAK (Waters Associates) round them (40 g), eluting with 10 * acetonitrile in water 55 kPa.

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

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

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

J = 6.0 Hz).

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

Analysis for C ₃ H ₂₂ N ₉ O 2 S. H, 6.10; N, 7.07. Found: C, 54.32; H, 6.03; N, 6.99.

Example 7

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

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

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

NMR (DMSO-d 6) δ: 1.15 (3H, d, J = 5.8 Hz), 2.62 (3H, s), 3.2 to 4.4 (5H, m), 4.45 ( 3H, s), 4.60 and 4.82 (1H each, ABq, J = 9.2 1-Iz), 5.30 and 5.46 (1H each, ABq, J = 12.8 Hz) and 7 6 to 8.9 (7H, m).

IR (KBr technique) gamma: 400, 1750 and 1590 cm @ -1

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

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

The reaction mixture was filtered through celite and the catalyst was washed with ether (2 x 10 mL). The combined filtrate and wash solution was extracted with ether (2 x 40 mL) and lyophilized to give a yellow solid which was purified on a C 4 g BONDAPAK (Waters Associates) column (20 g), eluting with 5% acetonitrile in water at 55 ° C. kPa.

Each 15 mL fraction was assayed by high pressure liquid chromatography and the fractions having a 11 11 affinity absorption at λ _max 300 nm were combined, and lyophilized to afford 141 mg (30% yield) of the title compound as a yellow solid.

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

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

IR (KBr) _ymax: 3400, 1750 and 1580 cm h UV lambda _max (H ₂ O): 296 nm (epsilon = 8014).

Example 8

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

OH

COjpNB

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

To a solution of Compound 9 (525 mg, 1.1 mmol) in methylene chloride (20 mL) was added methyl fluorosulfonate (0.27 mL, 3.3 mmol). The reaction mixture was stirred at room temperature for 5 minutes.

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

Then, to a solution of compound 10 in 100 mL of tetrahydrofuran and 100 mL of ether was added 100 mL of pH 7.0 buffer, followed by the addition of 500 mg of 10% palladium on carbon. The mixture is hydrogenated at 20 psi for 45 minutes on a Paar shaker.

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

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

Each 15 ml fraction was assayed by high pressure liquid chromatography, and fractions with ultraviolet absorption of lambda _max 300 nm were pooled and lyophilized. This afforded 145 mg (48% yield) of the title compound as a pale yellow powder.

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

IR (KBr) gamma: 3400, 1750 and 1585 cm

UV λ _max (H 2 O) ^: nm (epsilon = 7500).

Analysis for ^{C H} i5 I8 ° 4 ^N 2 ^S 2 • ^2H 2 ° Calculated: 46.15% C 5.64% H 7.17% N 16.41% S, 46.50 Found% C, 5, H, 26%; N, 7.13; S, 16.20.

Example 9

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

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

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

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

IR (KBr technique): _νm : 3400, 1750 and 1600 cm ^-1 .

* IÍIQA

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

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

The mixture was filtered through celite and the catalyst was washed with water (2 x 15 mL). The combined filtrate and washings were extracted with ether (3 x 100 mL) and lyophilized to give a yellow amorphous powder which was purified on a C 4 g BONDAPAK (Waters Associates) column (30 g) and eluted with 10% acetonitrile in water under pressure 55 kPa.

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

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

IR (KBr) gamma ^^^: 3500, 1750 and 1590 cm L UV _.lambda.max ^(Hjo): 296 nm (epsilon = 8411).

Anal. H, 5.67; N, 12.06; N, 9.50. Found: C, 49.93; H, 5.94; N, 11.46. % S.

Example 10

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

OH

Pd / C, H ₂

Ψ

3- (2,3,4-trimethylthiazol-5-yl-methanthio) -6alpha-E1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

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

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

IR (KBr) 3400, 1770 and 1690 cm @ -1

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

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

The mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (3 x 100 mL) and lyophilized to give a yellow powder which was purified on a C-4 BONDAPAK (Waters Associates) column (40 g), eluting with 10% acetonitrile in water at 55 pressure. kPa.

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

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

IR (KBr technician? 9 ^ama max ^: 3400, 1750 and 1580 cm @ -1)

UV λ _max λ 0: 297 nm (epsilon = 8,994).

Analysis for C-1. % H, 6.09; N, 7.79. Found: C, 47.96; H, 5.83; N, 7.89.

Example 11

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

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

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

To a solution of compound 7 in 140 mL of tetrahydrofuran and 120 mL of ether was added 140 mL of pH 7.0 buffer followed by 650 mg of 10% palladium on carbon. The reaction mixture was hydrogenated at 20 psi on a Paar shaker for 35 minutes.

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

Each 15 ml fraction was assayed by high pressure liquid chromatography, the ultraviolet absorption fractions at lambda _max 300 nm were combined and lyophilized to afford 23 mg (5% yield) of the title compound as a yellow amorphous solid.

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

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

IR (KBr) gamma: 3400, 1740 and 1580 cm \ ³ max.

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

Example 12

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

3- [-1- (RS) -methyl-N-methylpyridin-3-yl-methanthioyl-6-a- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-ene- 2-carboxylate

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

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

IR (KBr) ^{n tmax: 2400,} 1770 and 1190 cm

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

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

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

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

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

IR (KBr) ~ 'Nu _3χ: 3400, 1750 and 1590 cm ^-1.

UV λmax (H10): 296 nm (epsilon = 7573). max 2

Analysis for 37 ^ ^{Η Ν} 20 ° 4 ⁸ 2 1 ^^^ ^{2Η 2θ} calculated 54.38% C, 5.77% H 7.46% N Found 54.39% C, 5.98% H, 7 , 68% N.

Example 13

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

₄₃ COjpNB

4!

3- (N-methyl-1H-benzylimidazol-4-ylmethanthio) -6alpha-1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

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

The reaction mixture was concentrated to about 15 mL in vacuo.

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

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

IR (KBr) _m ax ^{b y:} 500, 1770 and 1700 cm ^1st

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

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

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

Each 15 ml fraction was assayed by high pressure liquid chromatografii.a fractions having an ultraviolet absorption at .lambda _χ 300 nm were collected and lyophilized to yield 305 mg (43% yield) of the title compound as a pale yellow amorphous solid.

NMR (DMSO) δ: 1.40 (3H, d, J = 7.0 Hz), 2.9-3.4 (3H, m), 3.98 (3H, s), 4.0-4

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

IR (KBr) ^{n Ymax:} 400, 1760 and 1590 cm \

UV λmax (H 2 O): 299 nm (epsilon = 8,807).

Analysis for ^C 21 ^H 23 ^N 4 S 3 ° 1 ^{'4 4H} 2 ° Calculated 57.25% C 5.94% H 9.54% N 7.28% S, 56.66 Found% C, 5 H, 70%; N 9.49;

Example 14

Preparation of 3- (2-methyl-N-methylpyridin-3-ylmethanthio) -6alpha- [4- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2.0) hept-2-en-2- carboxylate

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

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

NMR (CDCl3) of 17 delta: 1.20 (3H, d, J = 7Hz), 2.82 (3H, s,), 4.36 (3H, s), 4.16 (2H, m), 4.60 (2H, s), 5.20 (1H, m), 5.42 (2H, q, J = 14Hz), 7.80 (2H, d, J = 8Hz), 8.04 ( 1H, dd,

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

IR (KBr technology) ⁿ _{ma ma} x ^! 3,500 and 1,765 cm ⁹ .

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

To a solution of 1.10 g (1.88 mmol) of compound 17 in 80 mL of tetrahydrofuran and 80 mL of ether was added 80 mL of pH 7.0 buffer followed by 800 mg of 10% palladium on carbon. The mixture is hydrogenated at 20 psi on a Paar shaker for 40 minutes.

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

NMR (D ₂ O) δ: 1.28 (d, 3H, J = 7Hz), 2.86 (3H, s), 3.20 (2H, dd, J = 10Hz, 3.5Hz), 3.42 (1H, dd, J = 5.4 Hz, 3.5 Hz), 4.20 (3H, m), 4.32 (3H, s), 4.35 (2H, s), 9, 88 (1H, dd, J = 7.2Hz, 6.5Hz), 8.5 (1H, d, J = 8Hz) and 8.7 (1H, d, J = 8Hz).

IR (KBr) _ymax: 3400, 1760 and 1590 cm L UV _{.lambda.max (H2} 0): 298 nm (epsilon = 8391).

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

Example 15

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

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

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

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

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

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

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

Example 16

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

(4R, 5R, 6S) -3 - [[(1,33-Dimethyl-pyridinium-4-yl) -methyl] -thio] -6- [1- (R) -hydroxy-ethyl] -4-methyl-7-oxo-1-azabicyclo [ 3,2,0] Hept-2-ene-2-carboxylate (16B)

16A R = H 16B R = CH ₃

A. Preparation of 4-hydroxymethyl-3-methylpyridine

Boekelheid procedure (V. Boekel

1) h ₂ o ₂

- 2) Ac ₂ O

3) H ₃ O ®

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

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

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

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

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

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

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

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

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

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

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

To this mixture was added dropwise a solution of 4-hydroxymethyl-3-methylpyridine (4.0 g, 0.0325 mol) in 100 mL of anhydrous tetrahydrofuran and then freshly distilled thiol acetic acid (4.64 mL, 0.065 mol).

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

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

¹ H NMR (CDCl 3): 8.40, 7.20 (ABq, J = 5Hz, 2H), 8.37 (s, 1H), 4.08 (s, CHp, 2.35 (s, CH ₃ )) , 2.32 (s, CH _3); IR (oil) 1695 cm ^-1).

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

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

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

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

4.17 (s, _CH2), 2.53 (s, CH), 2.36 (s, CH); IR (oil) 1700 cm ^-1

D. Preparation of (5R, 6S) -3 - ([(1,3-dimethylpyridinium-4-yl) methyl] thio) -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.1] 2, θ] hept-2-ene-2-carboxylate

CH ₂ SAc

CH,

1) NaOH / H ₂ O

OH

I ch ₃

2) io-γ-ορ (οφ) ₂ CO ₂ pNB

OH

3) H ₂ , Pd / C

CH,

WHAT,

Nx /

CH,

To an ice-cooled nitrogen purged solution of sodium hydroxide (0.324 g, 0.008 mol) in 10 mL of water was added thioacetate (1.40 g, 0.004 mol) and the mixture was stirred at 0 ° C under nitrogen for 1 hour. The pH is then adjusted to 7.2-7.3 with a 10% aqueous potassium dihydrogen phosphate solution and the solution is added dropwise to an ice-cooled enolphosphate solution (1.45 g,

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

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

The aqueous phase was separated and the remaining solvents were removed in vacuo. The resulting aqueous solution was applied to a reverse phase column (C18 g Bondapak) which was eluted with water followed by 10% acetonitrile in water.

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

Lyophilization gave pure compound 16A (0.25 g, 57%) as a yellow solid. ¹ HNMR (θ ₂ Ο) δ 8.55 (s, 1H), 8.53, 7.96 (ABq, J = 6.8 Hz, 2H), 4.30-3.99 (m, 2H), 4.27 (s, 5H), 3.35 (dd, = 2.8 Hz, J ₂ - 6.0 Hz, 1H), 3.05 (d, J = 8.8 Hz, 2H), 2, 58 (s, 3H), 1.23 (d, J = 6.3Hz, 3H); IR (KBr) 1755, 1590 cm; UV (phosphate buffer, pH 7) 295 nm (epsilon 7 180).

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

3) H ₂ , Pd / C

To an ice-cooled solution of bicyclic ketone (0.906 g, 0.0025 mol) in 10 mL acetonitrile was added diphenylchlorophosphate (0.544 mL, 0.00263 mol), diisopropylethylamine (0.457 mL, 0.00263 mol) and 4-dimethylaminopyridine (0, 3 mg).

After 50 minutes, the reaction mixture was diluted with cold ethyl acetate and then washed with cold water and brine. The organic phase was dried (Na 2 SO 4) and evaporated at room temperature to give the enol phosphate as an off-white foam. This foam is taken up in 20 ml of tetrahydrofuran, cooled to -30 ° C under a nitrogen atmosphere and then treated with an aqueous solution of thiolate [prepared from 0.3 g of sodium hydroxide (7.5 mmol) and 1.3 g of thioacetate]. (3.76 mmol) in 10 mL of water].

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

After hydrogenation at 310 kPa for 1 hour, the mixture is filtered through Celite and the aqueous phase is separated and concentrated in vacuo. The resulting solution was applied to a reversed phase (C _1O Bonda1 by then), which was eluted with water.

Lyophilization of the appropriate fractions gave 1.2 g of a yellow solid. This was again chromatographed (eluting with water to 4% acetonitrile and water) to give the pure material after lyophilization. 16B (0.250 g, 28%) as a yellow solid.

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

J = 7.6 Hz, 3H) IR (KBr) 1750, 1595 cm -1; UV (phosphate buffer, pH 7) 292 nm (epsilon 7,930).

Example 17

Preparation of (5R, 6S) -3-f [(1,2-dimethylpyridinium-4-yl) methyl] thio] -6-L1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.1] o] Hept-2-ene-2-carboxylate (17A) (4R, 5R, 6S) -3-LL (1,2-dimethylpyridinium-4-yl) methyl] thio] -6- [1- (R) - hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate (17B)

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

The resulting mixture was stirred at room temperature for 30 minutes and then boiled under reflux for 2 hours. An additional 1.2 g (0.03 mol) of lithium aluminum hydride was added portionwise and refluxed for 1 hour.

An additional 1.2 g (0.03 mol) of lithium aluminum hydride was added portionwise and refluxed for 1 hour. The reaction mixture was then cooled to 0 ° C and 3.75 ml of water, 3.75 ml of a 15% aqueous sodium hydroxide solution were added, followed by 11.25 ml of water.

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

Obtained (7.7 g, 67%) as a yellow oil. ³ H NMR (CDCl ³ ) δ 8.30, 7.10 (ABq, J = 5 Hz, 2H), 7.17 (s, 1H), 5.42 (s, -OH), 4.70 (s, CHp, 2.50 (s, CH- ¹⁾

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

ch ₂ sac ch ₃

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

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

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

The filtrate was evaporated and the residual oil was chromatographed (silica gel / ethyl acetate-hexane 1: 1) to give thioacetate (8.87 g, 79%) as a yellow oil. ⁴ H NMR (CDCl3) δ 8.45,

7.03 (AB q, J = 5 Hz, 2H), 7.08 (s, 1H), 4.04 (s, _CH2), 2.55 (s, CH₃), 2.39 (s, CH) IR (oil) 1695 cm ^1st

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

4-Acetylthiomethyl-2-methylpyridine (1.368 g, 0.0075 mol) was added in one portion to 15 ml of ice-cooled nitrogen purged 1 N sodium hydroxide solution. After stirring at 0 ° C for 15 min, the reaction mixture was washed with ether (2 x 5 mL), neutralized with concentrated hydrochloric acid, and extracted with methylene chloride (3 x 10 mL).

Evaporation of the methylene chloride solution gave the thiol (0.89 g, 96%) as a yellow oil which gradually rose on standing. 1 H NMR (CDCl 3) δ 8.43, 7.37 (ABq, J = 5 Hz, 2H),

7.43 (s, 1H), 3.63 (d, J = 7.5 Hz, CH _2>, 2.55 (s, _CH3), 1.81 (t, J = 7.5 Hz, SH ).

D. Preparation of allyl- (5R, 6S) -3 - [- [(2-methylpyridin-4-yl) methyl] thio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [b] 3.2 of hept-2-ene-2-carboxylate

OH

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

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

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

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

%) as a white solid.

¹ H NMR (CDCl 3) 8.45, 10.09 (ABq, J = 5 Hz, 2H), 7.15 (s, 1H), 6.25-5.80 (m, 1H),

5.60 to 5.20 (m, 2H), 4i, 82 to 4.68 (m, 2H), 4.55 to 4.05 (m, 2H), 3.97 (s, 2H), 3, 16-2.93 (m, 3H), 2.55 (s, 3H), 1.84 (broad s, 1H), 1.32 (d, J = 6 Hz, 3H); IR (oil) 1,777,

695 cm ^-1 .

E. Preparation of (5R, 6S) -3- [[(1,2-dimethylpyridinium-4-yl) methyl] thio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabioyklo [3.2.1.0 &lt; 2,7 &gt; o] hept-2-ene-2-carboxylate

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

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

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

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

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

^X H-NMR (D ₂ O) delta 8.58, 7.83 (AB q, J = 6.4 Hz, 2H), 7.87 (s, 1H), 4.32 to 3.95 (m, 2H); 4.22) s, 2H), 4.17 (s, 3H), 3.32 (dd, = 2.6 Hz, J ₂ = 6.1 Hz, 1H), 3.06 to 2.93 (m 2H), 2.74 (s, 3H), 1.22 (d, J = 6.4 Hz, 3H); IR (KBr) 1757, 1590 cm ^-1 ; UV (phosphate buffer, pH 7.4) 296 nm (epsilon 7,446).

F. Preparation of allyl- (4R, 5R, 6S) -3- (2-methylpyridin-4-yl) methyl] thio-6- [beta] - (R) -hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo [3,2,0] hept-2-ene-2-carboxylate

A solution of the alpha-diazoester (1/50 g, 0.00508 mol) in 12 ml of ethyl acetate and hexane (3: 1) was heated under gentle reflux under a nitrogen atmosphere, then 0.020 g of rhodium octanoate was added in one portion. Nitrogen evolves rapidly for about 5 minutes and after boiling for a further 10 minutes the reaction is complete (thin layer chromatography).

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

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

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

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

The resulting gum was chromatographed on silica gel. Elution with ethyl acetate removed impurities and subsequent elution with acetonitrile gave the product (1.04 g, 53%) as a light yellow foam.

¹ H NMR (CDCl ₃ ) δ 8.43, 7.07 (ABq, J = 5Hz, 2H), 7.10 (s, 1H), 6.20-5.75 (m, 1H), 5.51 to 5.29 (m, 2H), 4.81 to 4.69 (m, 2H), 4.29 to 4.03 (m, 2H), 3.96 (s, 2H), 3.35 to 3 1.05 (m, 2H), 2.53 (s, 3H), 2.16 (broad s, 1H), 1.33 (d, J = 6.3 Hz, 3H), 1.22 (d, J = 7.3 Hz,

3H); IR (oil) 1,770, 1,705 cm ^-1 .

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

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

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

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

Dilution of the filtrate with ether gave an additional 0.041 g of crude product. The combined solids were dissolved in a small amount of phosphate buffer pH 7.4 (0.05M) and loaded onto a reverse phase column (C ^ g Bondapak).

Elution with water followed by 2% acetonitrile and water gave lyophilized compound 17B (0.413 g, 76%) as a yellow solid. ¹ H NMR (D ₂ O) δ 8.55, 7.76 (ABq, J = 6.3 Hz, 2H), 7.81 (s, 1H), 4.4-3.7 (m, 2H), 4.19 (s, 2H), 4.16 (s, 3H), 3.47 to 3.14 (m, 2H), 2.73 (s, 3H), 1.24 (d, J = 6, 4 Hz, 3H), 1.16 (d, J = 7.3 Hz, 3H); IR (KBr) 1750, 1595 cm -1 UV (phosphate buffer, pH 7.4) 293 nm (epsilon 7 170).

Example 18

Preparation of (5R, 6S) -3- (6-Dimethyl-pyridin-2-yl) -methylthio] -6- (1R-hydroxyethyl) -4R-methyl-7-oxo-1-azabicyclo [3] 2,0} hept-2-ene-2-carboxylate

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

CH ₃

O '' Ν '

O

SCCH,

MeOTf

-?

ether

HCl, 6N

Δ>

ch ₃

A solution of (6-methylpyridin-2-yl) methylthioacetate (1.0 g, 5.52 mmol) in anhydrous ether (5 ml) was treated with methyltrifluoromethanesulfonate (0.74 ml,

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

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

Appropriate fractions were combined and lyophilized to give a white powder, 1.43 g, 85.4%, IR (KBr) gamma,: 2565 (SH, 1626 (pyridinium), 1585 (pyridinium cm; UV (H_0)) lambda 278 inax of rnax (epsilon 7,355);

Analysis: calculated for C ₈ H ₁₂ NO ₃ S ₂ F ₃ :

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

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

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

O

1- (PhO) ₂ PCl

2- NEt (iPr) ₂ 3-Thiol

4-Net (iPr) ₂

H _2, Pd / C Buffer 7.0 ether

THF

coo

To a cold (5 C) solution of (5R, 6S) -p-nitrobenzyl-3,7-dioxo-6- (1R-hydroxyethyl) -4RS-methyl-1-azabicyclo [3.2.1] heptane-2-R- of carboxylate (1.11 g, 3.06 mmol, R / S: 86/14) in anhydrous acetonitrile (90 mL) were added diphenylchlorophosphate (0.68 mL, 3.3 mmol) and diisopropylethylamine under nitrogen atmosphere over 10 minutes. (0.57 mL, 3.3 mmol). The cold (5 ° C) mixture was stirred for 1 hour, cooled to -30 ° C, and a solution of (1,6-dimethylpyridinium-2-yl) methylthiol trifluoromethanesulfonate salt (1.03 g, 3.4) was added simultaneously over 15 minutes. mmol) in anhydrous acetonitrile (2 mL) and diisopropylethylamine (0.59 mL, 3.4 mmol).

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

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

The two phases were separated, and the aqueous phase was washed with ether (2 x 20 mL) and concentrated at -23 ° C under high vacuum to 15 mL and applied to the top of a reverse phase column (PrepPak C-18). Elution with 4% acetonitrile in water gave 0.23 g of the title compound mixed with potassium sodium diphenylphosphate (24% mole) after lyophilization of the appropriate fractions. Further purification on a reverse phase column (2.5 x 14 cm, PrepPak C-18) with water (400 mo) and 10% acetonitrile in water (200 ml) as eluent gave a yellow powder after freeze-drying of the appropriate fractions. 17 g 15.3% IR (KBr) gamma _max : 1750 (C = 0 beta-lactam), 1625 (pyridinium), 1600 (C = 0 carboxylate) cm ^-1 ; ¹ HNMR (D ₂ ) δ: 1.12 (d, J = 7.2 Hz, CH 8 on C-4), 1.24 (d, J = 6.4 Hz, CH 2 CHOH) 2.80 (s, CH3 on C-6 pyridinium), 4.18 (CH3 on N pyridinium), 4.41 (center of AB quartet, CH3S),

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

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

Example

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

COO

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

MeOTf

-γ ether

1- (PhO) ₂ POCl

2 NEt (iPr) ₂

3-thiol

4-Net (iPr) ₂

TfO © (PhO) ₂ PO ₂ o

To a cold (5 ° C) solution of (5R, 6S) p-nitrobenzyl-6- (1R-hydroxyethyl) -3,7-dioxo-1-azabicyclo [2.2.0] heptane-2R-carboxylate (2.14 g, 6.14 mmol) in anhydrous acetonitrile (18 mL) was added diphenylchlorophosphate (1.37 mL, 6.6 mmol), diisopropylethylamine (1.15 mL, 6.6 mmol) under nitrogen atmosphere at such a rate that the temperature was maintained at 5 ° C (7-10 min) and 4-dimethylaminopyridine (6 mg, 0.05 mmol).

The mixture was stirred at 5 ° C for 1.5 hours and used as it is. For further progress, this mixture is referred to as solution A. A solution of (6-methylpyridin-2-yl) methylthioacetate (1.23 g, 6.8 mmol) in anhydrous ether (10 mL) is treated with methyltrifluoromethanesulfonate (0) under nitrogen. (85 mL, 7.5 mmol) and stirred at 23 ° C for 1.5 h.

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

This mixture is referred to as solution B for further processing. The cold (5 ° C) solution A is treated with solution B for 0.5 hours, maintaining the pH between 7.25 and 7.35 by dropwise addition of 4N sodium hydroxide solution.

The mixture was stirred for 0.5 h and poured onto the top of a reverse phase column (4.0 x 18 cm, PrepPak C-18). The column was eluted with 25-50% acetonitrile in water. Lyophilization of the appropriate fractions gave the title compound as a yellow powder, 2.82 g (51 * (PhO) ₂ PO ₂ , 49%, CF 3 SO 4),

80%; IR (KBr) gamma _max : 3700-3000 (OH), 1772 (C = O beta-lactam), 1700 (C = O ester),

625 (pyridinium), 1590 (pyridinium), cm ³ H NMR (DMSO, d-6) δ: 1.15 (d, J = 6.2 Hz, CH 3 CHOH), 2.84 (s, CH 2 on C-); 6 pyridinium), 4.16 (s, CH 3 and N pyridinium), 4.79 (s, SCH ₂ ),

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

B. (5R, 6S) 3-Q (1,6-dimethylpyridinium-2-yl) methylthio] -6- (1-hydroxyethyl) -7-oxo-1-azabicyclo [2.2.0] hept-2-ene -2-carboxylate

COOpNB h ₂ , Pd / C

->

buffer

coo

To a solution of trifluoromethanesulfonate and diphenylphosphate salt (49:51) (5R, 6S) p-nitrobenzyl-3- (1,1,6-dimethylpyridinium-2-yl) methylthio] -6- (1R-hydroxyethyl) -7-oxo-1-azabicyclo [3,2, o] Hept-2-ene-2-carboxylate (0.87 g, 1.27 mmol) in wet tetrahydrofuran (50 mL) was added ether (50 mL), a potassium dihydrogen phosphate buffer and sodium hydroxide (0 mL). , 1 M) 40 ml, pH 7.0) and 10% palladium on carbon (0.87 g).

The mixture was hydrogenated for 2 hours at 23 ° C at 10 psi and filtered through Celite. The two phases were separated and the aqueous phase was washed with ether (2 x 15 mL), concentrated under high vacuum to 30 mL and poured onto the top of a reverse phase column (PrepPak C-18, 2.2 x 13 cm).

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

590 (c = O carboxylate) cm ^-1 : ¹ H NMR (D ₂ O) δ: 1.25 (d, J = 6.4 Hz, CH ₃ CHOH), 2.82 (s, CH ₃ on C-6) pyridinium), 3.12 (dd, J = 9.2 Hz, J = 2.9 Hz, C-4), 3.39 (dd, J = 6.0 Hz,

J = 2.8 Hz, H-6), 3.7-4, 4 (CH ₃ CHOH, H-5, CH ₃ on N pyridinium), 4.48 (s, CH ₂ S), 7.6 to 8 4 (hydrogen on pyridinium); UV (HO) lambda: 279 (epsilon 9 628) with inflex at 296:?] ^ ¹ of max.

55.0 ° C (c 0.63, H 2 O); ⁼ .About.5 hours measured at 37 DEG C. in a pH 7.4 buffer at a concentration of 10 ^{* 4} m.

Example

Preparation of 3- [2- (N-methylpyridiniummethanethiol) -6alpha-1- (R) -diethoxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

CO ₂ pNB

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

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

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

247195 ⁸⁴ (2H, q, J = 7.0 Hz), 3.60 (2H, s), 5.28 (2H, s), 7.45 (2H, d, J = 9.5 Hz) and 8 18 (2H, d)

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

The ice bath was removed and the mixture was allowed to stir for 90 minutes. The precipitate is filtered off, washed with acetonitrile and the filtrate is concentrated to about 100 ml volume and then diluted with 800 ml ethyl acetate. The organic solution was washed with aqueous sodium bicarbonate solution, brine and dried (MgSO 4.

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

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

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

OSi + \ X> 2 ^pNB

COjpNB _ n ₂

5

Β. 1-p-Nitrobenzyloxycarbonyl-1-diazo-2-tert-butyldimethylsilyloxy-2-butene (5)

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

The methylene chloride was dried (MgSO 4), filtered and evaporated in vacuo to give 68 g (89% yield) of compound 5 as a yellow solid, m.p. Mp 54-55 ° C.

IR (KBr) gamma 2080 and 1695 cm @ 1 NMR of Compound (5) shows that Compound (5) is obtained as a 9: 1 E / Z mixture in olefinic position. NMR (CDCl3 of the predominant isomer) delta 0.15 (6H, s). ), 0.90 (9H, ε), 1.58 (3H, d, J = 7.0 Hz), 5.15 (2H, s), 7.30 (2H, d, J = 9.0 Hz) ) and 8.0 (2H, d, J = 9.0 Hz).

OSi + .OAc

OSÍ +

-NH 1 x CO ₂ pNB

T

->

N ₂

OSi + S ^H 3 ^N 2

X—

I (CO ₂ pNB J - NH O

O, C. 4beta-1-methyl-3-diazo-3-p-nitrobenzyloxycarbonyl-2-oxopropyl) -3alpha- [[- (R) -ter. butyldimethylsilyloxyethyl] azetidin-2-one

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

A solution of 106 g (0.27 M) of 5 in 200 ml of methylene (200 ml) was added dropwise over 90 minutes to the reaction mixture.

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

*

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

IR (KBr) gamma 2130, 1760 and 1720 cm @ 1 H at 360 MHz of compound 7 shows that compound T_ is a 2: 1 mixture of methyl isomers. NMR (CDCl 3) δ 0.3 to 0.6 (6H, 2s), 0.82 (9H, 2s), 1.05 to 1.15 (6H, m), 2.68 (0.88 H, q, J = 6.6 and 2.0 Hz), 2.88 (0.34 q,

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

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

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

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

IR (KBr) gamma 3400, 2135 and 1750 cm @ ^-1 . NMR at 360 MHz of compound 8 shows that compound 8 is obtained as a 2: 1 mixture at the 4-position of methyl. NMR (DMSO-d)?

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

3.90 (3H, m), 5.25 (2H, s), 7.70 (2H, d, J = 9.0 Hz), 8.05 (0.66H, s), 8.10 (0 34H, s) and 8.27 (2H, d, J = 9.0)

Hz).

OH

CO ₂ pNB

- =?

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

A solution of 14.0 g (35.86 mmol) of compound III and 70 mg of rhodium (II) octanoate in ethyl acetate was refluxed for 20 minutes under a nitrogen atmosphere. The mixture was evaporated in vacuo to give compound 9 as a foam. IR (CHCl 3) gamma 3400 and 1750 cm ³ . NMR at 360 MHz of compound 9 shows that compound 9 is obtained as a 2: 1 mixture in the 4-position of methyl. Nuclear Overhauser effect is used to determine the 4-methyl configuration. If H is irradiated. of the isomer, which is more, there will be an approximately 7% increase in the signal for 4-methyl protons, indicating the cis configuration of H 2 and 4-methyl.

Conversely, if the less isomer is irradiated, there is no desirable increase in the signal for 4-methyl, indicating a trans configuration and 4-methyl for the less isomer.

NMR (CDCl3) isomer of the more delta 1.24 (3H, d, J = 7.35 Hz), 1.40 (3H, d, J = 6.3 Hz), 2.40 (1H, m) 3.24 (1H, q, J = 6.6 and 7.2 Hz), 3.76 (1H, q, J = 8.0 and 2.2 Hz), 4.18 (1H, m),

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

NMR (CDCl3) isomer of less delta 1.0 (3H, d, H = 7.35 Hz), 1.40 (3H, d,

J = 6.3 Hz), 2.83 (1H, m), 3.25 (1H, q, J = 6.6 and 1.50 Hz), 4.14 (1H, q, J = 7.36) and 1.50 Hz), 4.67 (1H, s), 5.24 (1H, d, J = 6.3 Hz), 6.18 (1H, d, J = 6.3 Hz) and 7, 60 (1H, d, J = 8.5Hz), and 8.22 (1H, d, J = 8.5Hz).

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

F. p-Nitrobenzyl-3-diphenoxyphosphinyl-6alpha- [1- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate (10) )

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

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

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

IR (CHCl 3) gamma 3400, 1790 and 1720 cm ^-1 .

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

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

The Nuclear Overhauser effect is used to determine the 4-methyl configuration of compound 10. When irradiated with H 2, there was no signal increase for 4-methyl, indicating a trans configuration of H 1 and 4-methyl.

/ Xz

G. p-Nitrobenzyl-3-pyridin-2-yl-methanthyl] 6a, [1- (R) -hyloxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo [3,2.0] hept-2-ene 2-carboxylate (11)

To a cooled (-15 ° C) solution of 1.2 g (2 mmol) of phosphonate 10 in 10 mL of acetonitrile was added 390 mg (3 mmol) of diisopropylethylamine followed by 370 mg (3 mmol) of 2-mercaptomethylpyridine under nitrogen.

The reaction mixture was stirred at -15 ° C for 60 minutes and then at 0 ° C for a further 60 minutes. The reaction mixture was diluted with ethyl acetate, washed with ice water, brine and dried (MgSO 4). Evaporation of the solvents in vacuo afforded a yellow oil which was purified on a silica column.

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

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

H. 3- [2- (N-Methylpyridinium) methanothioyl-6-alpha-yl- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3,2,0) hepet-2- en-2-carboxylate (12)

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

The crude pyridinium salt was dissolved in TMF-ether pH 7 buffer (1: 1: 1, 100 mL each), followed by the addition of 600 mg of 10% palladium on carbon. The mixture is hydrogenated for 45 minutes at 20 psi on a shaker. The mixture was filtered through celite and the catalyst was washed with water (2 x 10 mL).

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

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

IR (KBr) Gamma 3400, 1750 and 1590 cm Lambda _max (H ₂ O) = 92 ^{nm (P} ^ l ⁿ ° 7081). NMR (D ₂ O) δ 1.13 (3H, d, J = 6.5 Hz), 1.23 (4H, d, J = 6.5 Hz), 3.18 (1H, m), 3, 45 (1 H, q,

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

Example 21

Preparation of 3- [2- (1,4-dimethylpyridinium methanthio) -6alpha- [1 (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-en-2 -carboxylate

3- [2- (1,4-dimethylpyridiniummethanethio) -6alpha- [* 1- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3,2,0) hept-2-enecarboxylate (13)

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

Gamma IR 3400, 1755 and 1600 cm \ UV .lambda.max (H _o 0) 300 nm (e 7600). NMR (D _o O) max zz delta 1.20 (3H, d, J = 6.7 Hz), 1.28 (3H, d, J = 6.7 Hz), 2.60 (3H, s) 3.4 to 3.5 (2H, m), 4.2 to 4.4 (4H, m), 4.52 (3H, s), 7.82 (1H, t, J = 6.5 and 4) 2 Hz), 8.32 (1H, d, J = 6.5 Hz) and 8.60 (1H, d, J = 4.2 Hz).

Example 22

Preparation of 3- [4- (1-methylpyridinoneunethanthiq) -6alpha-f- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate

OH

1) HS

2) CF ₃ SO ₃ CH ₃

3) Pd / H ₂ // 7 COF ©

] -ch ₃

3- [4- (1-methylpyridinium methanthioj) -6alpha- [1- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate ( 14)

This compound was obtained as a yellow powder in 15% yield from compound 10 in the same manner as described in Example 20. IR (KBr) gamma 34104 1750 and 1650 cm. UV λ _max (H ₂ O) 293 nm (epsilon 7,295). NMR (D ₂ O) δ 1.15 (3H, d, J = 6.5 Hz), 1.20 (3H, d, J = 6.5 Hz), J = 6.5 Hz), 3.20 (1H, m), 3.45 (1H, q, J = 6.0 and 2.0 Hz), 4.11 (1H, q, J = 8.0 and 2.0 Hz),

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

Example 23

Preparation of 3- [3- (1-methylpyridinium) methanthio] -6alpha- [1- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene 2-carboxylate

3- [3- (1-Methyl-pyridinium-methanthio) -6alpha-Q1- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate ( 15)

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

IR (KBr) gamma 3420, 1750 and 1610 cm ^-1 . UV λ _max (H ₂ O) 295 nm (epsilon 8 750).

NMR (D ₂₀ ) δ 1.10 (3H, d, J = 6.9 Hz), 1.25 (3H, d, J = 6.9 Hz), 1.27 (1H, m), 1, 43 (1H, q, J = 6.2 and 1.8 Hz), 4.1 to 4.35 (4H, m), 4.39 (3H, s), 8.0 (1H, t, J = 8.5 and 6.2 Hz), 8.45 (1H, d, J = 8.5 Hz), 8.70 (1H, d, J = 6.2 Hz) and 8.82 (1H, s) .

For C 6 proHjqNjO :S: S 1/2 pro HHjO: C 51.90, H 6.36, N 7.12; Found C 51.92, H 5.71, N 6.88.

Example 24

Preparation of 3- [3- (1,2, -dimethylpyridinium) methanesulfonyl-6α- [1- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabioylclo (3,2,0) hept-2 -en-2-carboxylate

2) CF 3 SO 3 CH 3

->

3) Pd / H ₂

3- [3- (1,2-Dimethyl-pyridinium-methanesulfonyl) -6-alpha - [[1- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabioylclo (3,2,0) hept-2-en-2 carboxylate (16)

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

IR (KBr) Gamma 3410, 1750 and 1600 cm UV lambda _max (H ₂ O) ² ≤ 6 nm ^(P ^ -i ⁿ ° 500 ®).

NMR (D ₂ O) δ 1.25 (3H, d, J = 6.5 Hz), 1.30 (4H, d, J = 6.5 Hz), 2.95 (3H, s), 3, 40 (1H, m), 3.50 (1H, q, J = 6.2 and 1.8 Hz), 4.2-4.4 (4H, m), 4.35 (3H, s), 7.82 (1H, t, J = 8.5 and 6.3 Hz), 8.40 (1H, d, J = 8.5 Hz) and 8.72 (1H, d, J = 6.3 Hz) ).

Example 25

Preparation of 3- (2,4-dimethyl-1,2,4-triazolium) methanthio-6alpha-Q1- (R) -hydroxyethyl-4-beta, 1'-methyl-1-oxo-1-azabicyclo (3,2,0) of hept-2-ene-2-carboxylate ch ₃

2) CF ₃ SO ₃ CH ₃

3) Pd / H ₂

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

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

IC (KBr) gamma 3420, 1756 and 1725 cm -1 UV λmax (H ₂ O) 291 nm (epsilon 7 850).

NMR (D ₂ O) δ 1.15 (3H, d, J = 6.3 Hz), 1.22 (3H, d, J = 6.3 Hz), 3.35 (1H, m), 3, 48 (1H, q), J = 6.0 and 1.8 Hz), 3.90 (3H, s), 4.05 (3H, s), 4.2 to 4.4 (4H, m), 8.80 (1H, s).

Example 26

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

3- [2- (1,3-dimethylimidazolium methanthio) -6-α- [1- (R) -hydroxyethyl] -4-beta-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-en-2- carboxylate (18)

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

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

NMR (D ₂ O) δ 1.10 (3H, d, J = 6.3 Hz), 1.25 (3H, d, J = 6.3 Hz), 3.30 (1H, m), 3, 42 (1H, 1, J = 6.0 and 2.2 Hz), 3.85 (6H, s), 4.2 to 4.6 (4H, m), and 7.40 (2H, s).

Example 27

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

Example

C.

27a

-CH

27b

-ch ₂ ch ₂

N-CH

27c

-CH

CH,

27d

-CH ^CH \ ®

27e

-ch ₂ ch ₂ -

O ₂ ch ₂ -ch ₂ -ch ₃

CH, \ ®

-p

CH,

27h

-CH _n 247195

27i

_N -CH IT

CH,

I® .N.

O

CH *

27j

CHg - r - N - CH 3 - CH ₃

27k

-CH _n

- <j ®r

CH,

271

CH, 3-CH-O ^? ch ₃

7tn ® /

CH,

27n

-CH ~ 27o

-CH-

CH ₃ (mixture of 3 possible

27p

Τ ^- ί s' s ^ n-ch ₃ coo ^fc

CH,

27q

CH,

Example

C.

CH,

27r

-CH ". ©

27s

-CH _n

27t

-CH-ch ₂ -coo

CH,

I ©

-AND

Ν — N, © ch ₃

N-CH, N-N

CH,

Example 28

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

CH,

28b \ ©

-O

28c - .beta. @ 4 ^{H4 CH2 CH2 CH2}

Example

C.

28d

-CH-CH 11> ^H 3

28e

28f ^CH3 XJtc ^C H ^H 3 ³

N-

CH,

28g

CH,

-AT

28h

28i

28j

28k

281

8m i mixture of N-p-N CHg isomers)

CH,

N

AND! © xN-CH

COO

CH,

I - N

CH,

CH, ch ₂ -coo ch ₃

I ©

N,

N - N (mixture of 3 possible

CH,

Example

E.

O ^{% s}

28n

-CH

N-N-CH, > N = N

CH,

Example 29

When the ketone intermediate is replaced by an equimolar amount of the corresponding 1-alpha-methyl intermediate, the above carbapenem product is obtained.

Claims (7)

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