AU734948B2 - Cephalosporin derivatives - Google Patents

Description

1 CEPHALOSPORIN DERIVATIVES 1. Field of the Invention The present invention is directed to new cephem derivatives represented by the general formula (X)m

COOM

0S

S

S

wherein Q is an optionally substituted pyridinium group connected to the sulfur atom via a ring carbon atom; X is halogen; Y is hydrogen or halogen; A is

CO

2 H, PO 3

H

2

SO

3 H or tetrazole; L 1 is a furylene group, a thienylene group, a

C

2

-C

10 alkylene group, or a C 2

-C

10 alkylene group interrupted by one or more groups independently selected from vinyl, S, SO, SO 2

SO

2

NH;

S

0

AN/

H

H

or N

O

n is 0 or 1; M.is hydrogen, a negative charge or a carboxyl-protecting group; m is 0 or an integer selected so as to make the molecule as a whole electrically neutral; and X G is a counter anion; or a pharmaceutically acceptable salt and/or a prodrug thereof.

The derivatives are gram-positive antibacterial agents especially useful in the treatment of diseases caused by methicillin-resistant Staphylococcus aureus (also referred to below as MRSA or methicillin-resistant S. aureus).

W:\SPEC53652-08.doC WO 98/23621 PCT/US97/21785 2 2. Description of the Prior Art The literature discloses a vast number of cephem derivatives having a wide variety of C-3 and C-7 substituents.

With respect to the C-7 substituents of the present invention, U.S.

Patent 3,345,366 discloses cephem derivatives of the type 0 II o

R

2 -C-CH2

H

Z-CH

2 -C-N s

R

N CH 2

-A

0 CO2M wherein R, is hydrogen or chloro, R 2 is hydroxy or amino, Z is oxygen or sulfur, A is acetoxy or N-pyridinium and M is hydrogen, pharmaceutically acceptable non-toxic cations or an anionic charge when A is N-pyridinium. This approach was also discussed in Antimicrob. Ag.

Chemother. Meeting, 1968, pgs 109-114 (Hobby, and JP 50083383.

EP 638,574 Al discloses cephem derivatives of the general formula yjr

R

II

CH

(CH

2 )n Z X-CH2--ILN

COOH

WO 98123621 WO 98/362 1PCTIUS97/21785 3 wherein X absent, -SO2_1 -NH-; Y -CH, N-; Z halogen, -OH, C,-C 5 0-alkyl, -OCH 2

CONH

2

-OCONH

2

I-OSO

2

NH

2 I -OCH 2 CN, -NH 2 either as such or substituted with C 1

-C

6 alkyl radicals, -NHCOCH 3

NHSO

2

CH

3 -NHS0 _Q CH3 amides of C-C 4 linear acids, amides of benzene and toluene derivatives, -NO 2

-NO,

-CHO, -CH 2 OH, -COOH, -SH, -SOH, -S2H -SO 3 H, -S-alkyl where the alkyl residue is CI-C 3

-CF

3 R -OH, Cl-C 5 ,-O-alkyl with the alkyl residue possibly containing halogens, acid functionalities either free or salified with alkaline or alkaline earth metals, basic functions such as -OCH 2

CH

2

NH

2

-OCH

2

CH

2

NH-CH

3

OCH

2 m, p)-pyridinyl,

-OCH

2 CN, -OCH 2

CONH

2

OCH

2

SO

2

NH

2 n =0 to 4; A

-CH

2

SO

2

R

1 a structural group characteristic of cephalosporins such as -Cl, -OCH 3

,-_CH

2

OCH

2

NH

2

-CH

2

OCH

3

-CHY,-CH=CH-

CH ,CF3 -CO 2

R

2

-SO

2 R where R 2 is an alkyl or aryl radical WO 98/23621 PCT/US97/21785

N-N

-CH

2

-OCOCH

3

-CH

2 .N CH3

(CH

22

-N-CH

3

CH

2

-N

N-N

-CH2-S- N

N

CH

3 -CH=

CH

2

CH

3 N N OH CHTS N 1 0~

N

CHCHS

CH

3

-CH

2 -S CH 2

COOH

CH

3

CH

2

COOH

N-N

-CH2-S N- N

NH.

N

-CH2-S

N.

N

-CH

2 -S S

CH

3

N.

x\

N

-CH2-S S their pharmaceutically acceptable salts and their C 6 and C 7 epimers.

The C-3 substituents employed in the compounds of the present invention are known in the cephem art, but have not previously been combined with the C-7 substituents of the present invention. Applicants have discovered that the combination of C-3 and C-7 substituents provided in the compounds of the present invention unexpectedly gives the desired solubility, activity and toxicity profile needed for commercially viable anti-MRSA cephem products.

SUMMARY OF THE INVENTION The present invention provides a novel series of cephem derivatives of the general formula X Y 0A N

/-CH

2

-S-Q

COOM

wherein Q is an optionally substituted pyridinium group connected to the sulfur atom via a ring carbon atom; X is halogen; Y is hydrogen or halogen; A is

CO

2 H, PO 3

H

2

SO

3 H or tetrazole; L 1 is a furylene group, a thienylene group, a C2-C10 alkylene group, or a C2-C10 alkylene group interrupted by one or more groups independently selected from vinyl, S, SO, SO2, SO 2

NH,

O H or N- H O 10 0 n is 0 or 1; M is hydrogen, a negative charge or a carboxyl-protecting group; m is 0 or an integer selected so as to make the molecule as a whole electrically neutral; and X e is a counter anion; or a pharmaceutically acceptable salt and/or a prodrug thereof.

The compounds of formula I are antibacterial agents useful in the treatment of infections in humans and other animals caused by a variety of gram-positive bacteria, particularly methicillin-resistant S. aureus.

Also included in the invention are processes for preparing the compounds of formula I and pharmaceutical compositions containing said compounds in combination with pharmaceutically acceptable carriers or excipients.

W:\SPECI53652-98.doc The discussion of documents, acts, materials, devices, articles and the like is included in this specification solely for the purpose of providing a context for the present invention. It is not suggested or represented that any or all of these matters formed part of the prior art base or were common general knowledge in the field relevant to the present invention as it existed in Australia before the priority date of each claim of this application.

Throughout the description and claims of this specification, the word "comprise" and variations of the word, such as "comprising" and "comprises", is not intended to exclude other additives, components, integers or steps.

W:\anellespeci53652.doc WO 98/23621 PCT/US97/21785 6 DETAILED DESCRIPTION The present invention provides novel cephem derivatives of general formula I above which are antibacterial agents useful in the treatment of infectious diseases in humans and other animals. The compounds exhibit good activity against a wide variety of gram-positive microorganisms, e.g. S. pneumoniae, S. pyogenes, S. aureus, E. faecalis, S.

epidermidis and S. hemolyticus, and are particularly useful against strains of methicillin-resistant S. aureus.

The compounds of formula I are characterized by an optionally substituted pyridinium thiomethyl group of the type CH2-S Nat the 3-position of the cephem ring and a 7-substituent of the type o f 1 A-(fL )n S-CH 2

-C-NH-

wherein X is halogen; Y is hydrogen or halogen; A is CO 2 H, PO 3

H

2

SO

3

H

or tetrazole; L 1 is a furan group, a thiophene group, a C 2

-C

10 alkyl group, or a C 2

-C

10 alkyl group interrupted by one or more (preferably 1 or 2) groups independently selected from vinyl, S, SO, SO 2

SO

2

NH,

0

AN/

H

H

or N 0 and n is 0 or 1.

A preferred embodiment of the present invention comprises a compound of the formula X Y SH (X)m® 0 -N CH--S N-R COOM 4

IA

wherein X is halogen; Y is hydrogen or halogen; A is CO 2 H, P0 3

H

2

SO

3 H or tetrazolyl; L 1 is a furylene group, a thienylene group, a C 2

-C

10 alkylene group, or a C 2 -C10 alkylene group interrupted by one or more groups independently selected from vinyl, S, SO, SO2, SO 2

NH,

S.

S

0

AN-

H

or /N

O

n is 0 or 1; R 3 and R 4 are each independently selected from hydrogen or C 1

-C

6 alkyl; R 2 is hydrogen, NH 2 pyrrolidinyl, C 3

-C

6 cycloalkyl, C 1

-C

6 alkyl, C 2

-C

6 alkyl substituted by one or more substituents independently selected from OH,

NR'R

6 in which R 5 and R 6 are each independently hydrogen or Ci-C 6 alkyl,

CO

2 H, morpholinyl, morpholinyl quaternized by a C 1

-C

6 alkyl group, oxo, v halogen, SO 3 H, PO 3

H

2 imidazolyl, imidazolyl substituted by 1-2 Cl-C 6 alkyl -n S groups, tetrazolyl, tetrazolyl substituted by 1-2 C1-C 6 alkyl groups or N=CR 7 in which R 7 is a furan or thiophene ring optionally substituted by either -C0 2 H or W:\SPEC3652-98.doC

-SO

3 H, phenyl or phenyl substituted by 1-3 substituents independently selected from OH, NR 5

R

6 in which R 5 and R 6 are as defined above, CO 2 H, morpholinyl, morpholinyl quaternized by a C1-C6 alkyl group, oxo, halogen, SO 3 H, PO 3

H

2 imidazolyl, imidazolyl substituted by 1-2 C1-C 6 alkyl groups, tetrazolyl, tetrazolyl substituted by 1-2 C1-C6 alkyl groups or N=CR 7 in which R 7 is as defined above; M is hydrogen, a carboxyl-protecting group or a negative charge; X E is a counter anion; and m is 0 or an integer selected so as to make the molecule as a whole electrically neutral; or a pharmaceutically acceptable salt and/or a prodrug thereof.

Another preferred embodiment comprises a compound of the formula X Y IH 3

(X)

1S. N S R3 i A- )n D ,I A 0 N CH 2 -S N-R 2 S" COOM R4

IA

wherein X is halogen; Y is hydrogen or halogen; A is CO 2 H, P0 3

H

2

SO

3 H or tetrazolyl; L 1 is a furylene group, a thienylene group, a C2-C10 alkylene group or C2-C10 alkylene group interrupted by one or two groups independently selected from vinyl, S, SO, SO 2

SO

2

NH,

O H *N or /N H 0 n is 0 or 1; R 3 and R 4 are each independently selected from hydrogen or CI-C6 alkyl; R 2 is hydrogen, NH 2 pyrrolidinyl, C1-C6 alkyl, Ci-C6 alkyl substituted by SRA, one or two substituents independently selected from OH, NR 5

R

6 in which R and R 6 are each independently hydrogen or C1-C6 alkyl, C0 2 H, morpholinyl, 2 morpholinyl quaternized by a C1-C6 alkyl group, oxo, halogen, SO 3 H, PO 3

H

2 tetrazolyl, W:lSPECI53652-96.doc 9

CH

3 -N N-CH 3 or -N R 7 in which R 7 is a furan or thiophene radical optionally substituted by a -CO 2 H or

-SO

3 H group, phenyl or phenyl substituted by 1-2 substituents independently selectedrfrom OH, NR 5

R

6 in which R 5 and R 6 are as defined above, CO 2

H,

SO

3 H, P0 3

H

2 tetrazolyl,

CH

3 -N N-CH 3 or halogen; M is hydrogen, a carboxyl-protecting group or a negative charge; X

E

is a counter anion; and m is 0 or an integer selected so as to make the molecule as a whole electrically neutral; or a pharmaceutically acceptable salt and/or a prodrug thereof.

Particularly preferred groups of the formula X Y l A- (Lo) in the above-described compounds of formula I and formula IA include W:\SPEC53652-98.doc VO 98/23621 WO 98/362 1PCTIUS97/21785 C1 C0 2

H

c1 C1

O=SN

C0 2

H

C0 2

H

C1

CI

C0 2

H

S")

C0 2

H

C-

0=S C0 2

H

Cl1

HO

2 C N 0

H

C0 2

H

CI

-0 C0 2

H

CI

C0 2

H

SS s C1 C1

CO

2 H

HOC

C1 Ci

CO

2

H

H0 2

C~'

Particularly preferred groups of the formula N-

R

2 include WO 98/23621 PCTIUS97/21785

H

3

C

N-NH

2

CH

3 N NH

CH

3

CH

3

OH

0 C N L1' 0 S03H

CH

3

NH?

OH

CH

3

CH

3 00

H

3

C

CH

3

CH

3

OH

CO

2

H

CH

3 NH3

OH

CH

3

OH

C C02H

OH

O

CH

3 CH3 ~-CH3

CH

3 Specific preferred embodiments of the present invention include the following: WO 98/23621 WO 9823621PCTIUS97/21785 12 x Y

H

COOR'

wherein R' is hydrogen or a carboxyl-protecting group x Y

CI

Cl C0 2

H

H

3 C _j 1-[3-[N-(N-methyl) morpholino)I-prop-1-yl-4-[ [(6R)-trans-2-carboxy-8-oxo- 7 2 5 -dichloro-4-(2-carboxyethenyl)phenylthio)acetamido]5thia-l-..

azabicyclo[4.2.01-oct-2-en-3-yl]methylthiolpyridinium inner salt

CI

C0 2

H

CH

3 NNH H

CH

3 WO 98/23621 WO 9823621PCTIUS97/21785 13 2,6-dimethyl-1 [amino] [[(6R)-trans-2-carboxy-8-oxo-7- (2,5-dichloro-4-(2carboxyethenyl)phenylthio)acetamido]-5-thia--azabicyco4.2.O]-oct-2-en-3yllmethylthiolpyridinium inner salt cl C0 2

H

NH

2 1 -[amnino] [(6R)-trans-2-carboxy-8-oxo-7- [(2,5-dichloro-4-(2carboxyethenyl)phenylthio)acetamidol-5-thia--azabicyclo[420oct2en-3 yllmethylthiolpyridinium inner salt

(D

N Nn Cl C0 2

H

l-IN-pyrrolidino-4-[[(6R)-trans-2-carboxy-8-oxo-7-[ (2,5-dichloro-4-(2carboxyethenyl)phenylthio)acetamido]-5-thia-l-azabicyclo[420oct2en-3 yllmethylthiolpyridinium inner salt WO 98/23621 PCTIUS97/21785 14 Cl CH 3 C0 2

H

l-[methyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7- [(2,5-dichloro-4-(2carboxyetheny)phenylhio)acetamido5thialazabicyclol42Op]oct2en-3 yllmethylthiolpyridinium inner salt S

CH

3 Wf CI iN OH

CH

3 H0 2 C N 0

H

2,6 dimethyl-1-[2-hydroxy ethyl] [(6R)-trans-2-carboxy-8-oxo-7- dichloro-4-(2-[ [(1S)-(carboxy-l1-ethyl(amino)carbonyl)phenylthio) acetamido]-5-thia-1-azabicyclo[4.2.o] -oct-2-en-3-yllmethylthio] pyridirium inner salt WO 98/23621 WO 9823621PCT/US97/21785 c' C0 2

H

-NH 2 1 [amino] [(6R)-trans-2-carboxy-8-oxo-7- (2,5-dichloro-4-(carboxymethyl thio)phenylthio)acetamido]-5thia.1..azabicyclo[420]oct-2en3yllmethylthiolpyridinium inner salt 0

NH

2

CO

2

H

1- [aminocarbonyl methyll-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichloro-4 (carboxymethylthio)phenylthio)acetamido] -5-thia-1 -azabicyclo[4.2.0]-oct-2en-3-yllmethylthiolpyridinium inner salt C0 2

H

NH2

OH

WO 98/23621 WO 98/362 1PCTIUS97/21785 16 1-[2-hydroxy-3-amino-prop-1-yl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5dichloro-4-(2-carboxyethenyl)phenylthio)acetamido-5-thia-1 azabicyclo[4.2.O]-oct-2en-3-yl]methylthio]pyridinium inner salt

OH

C0 2

H

1-[3-hydroxy-4-carboxyphenyl]-4-[ [(6R)-trans-2-carboxy-8-oxo-7-[(2,5dichloro-4-(2-carboxyethenyl)phenylthio)acetamido] -5-thia-1 azabicycloll4.2.0]-oct-2-en-3-yllmethylthiolpyridinium inner salt

CI

C0 2

H

C~N~ C0 S03H l-[5-(sulfonylfuran-2-yl)-methylimine]-4-[ [(6R)-trans-2-carboxy-8-oxo-7- 2 ,5-dichloro-4-(2-carboxyethenyl)phenylthio)acetamido-5-thia.1 azabicyclo[4.2.0]-oct-2-en-3-yllmethylthiolpyridinium inner salt WO 98/23621 WO 9823621PCTIUS97/21785

CH

3 C .1 rNH 2

CH

3 C0 2

H

oxo- 7 -[(2,5-dichloro-4-(2-carboxyeth-1 -yl)phenylthio)acetamido]-5-thia-1 azabicyclo[ 4 2 .0I-oct-2-en-.3..yljmethylthiojpyridinium inner salt

CH

3 (in) ClC ciCH 3 N-CH3 C0 2

H

1,2 dimethy1-1H-imidazo-3)prop1y]l.4.[[(6R)-trans-2-carboxy-8-oxo- 7 2 5 -dichloro-4-carboxyphenylthio)acetamido]-5.thia.l-azabicyclo oct- 2 -en-3-yl]methylthiolpyridinium inner salt cl C0 2

H

CXT----OH

WO 98/23621 WO 98/362 1PCTIUS97/21785 18 1-[2-hydroxyethyl]-4-[[ (6R)-trans-2-carboxy-8-oxo-7- [(2,5-dichloro-4oct-2-en-3-yllmethylthiolpyridinium inner salt cI

CIS

C0 2

H

CH

3 (D GD H 3

CH

3

N-H

2,6-dimethyl-1-[3-(3-(1,2 dimethyl- 1H-imidazol-3)prop-1-yl]-4- [[(6R)-trans- 2-carboxy-8-oxo-7-[(2,5-dichloro-4azabicyclo[4.2.0] -oct-2-en-3-yl] methyithiolpyridinium inner salt

CI

C (D ri.,

OH

C0 2

H

1-[2-hydroxyethyll-4-[[(6R)-trans-2-carboxy-8-oxo-7- [(2,5-dichloro-4-((( 1-azabicyclo 4 2 .]-oct-2-en-3-yl]methylthiolpyridinium inner salt WO 98/23621 WO 9823621PCTIUS97/21785

OH

HO

2 N -A,0

H

l-[ 2 -hydroxyethyll-4-[[(6R)-trans-2-carboxy-8-oxo7[(2Sdichloro-4(2- 1 azabicyclo[4.2.0]-oct-2-en-3..yljmethylthiolpyridinium inner salt

S.

c,

CI

-S

C0 2

H

CH

3 (D H

CCH-

3 L 1- dimethyl-1H-imidazol-3-yl)prop-1 -yl]-4-[[(6R)-trans-2-carboxy-8oxo-7-[ 2 ,5-dichloro-4-(5-carboxy-2-thiophene)phenylthio)acetamidol-5.

thia-l-azabicyclo[4.2.-oct2en-3yllmethylthiolpyridinium inner salt (s Cl QI 3

NH

2

CH

3 WO 98/23621 WO 9823621PCTIUS97/21785 2,6-dimethyl-l-jlaminoj-4-[ [(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichloro-4-(2- (((carboxymethyl)amino)carbony1)ethenyl)phenythio)acetamido-5thia..1 azabicycloll 4 2 .O-oct-2-en-3-yllmethylthiolpyridinium inner salt

CCI

HO

2 N 0 H:2 H

CH

3

CH

3 2,6-dimethyl- 1-4 N-(N-methyl) morpholinol-4-[[(6R)-trans-2-carboxy-8-xo- 7 2 ,-dichloro-4-(2-(((carbonylmethyl)amino) carbonylethenyl)phenylthio)acetamido-5-thia.l-azabicyclo[4.2.0] oct-2-en- 3 -yllmethylthio]pyridinium inner salt -s o

NH

C0 2

H

CH3 (D CH3 CII I ,,!NCH3 CH3 2,6-dimethyl-1-[ l,2-dimethyl-1H-imidazol-3-yl)prop-1-y1].4[ [(6R)-trans-2carboxy-8-oxo-7-[(2,5-dichloro-4-(NcarboxymethyI aminosulfonyl)phenylthio)acetamido] -5-thia-l-azabicyclo[4.2.0] -oct-2-en-3yllmethylthiolpyridinium inner salt WO 98/23621 PCTIUS97/21785 21 or

S

C1 G

OH

CO

2

H

1-[2-hydroxyethyl]-4-[[(6R)-trans-2-carboxy-8-oxo-7-[(2,5-dichloro-4-(5carboxy-2-furan)phenylthio)acetamido]-5-thia- -azabicyclo[4.2.0]-oct-2-en- 3 -yl]methylthio]pyridinium inner salt; or pharmaceutically acceptable salts and/or prodrugs thereof.

To elaborate on the definitions for substituents of the formula I and formula IA compounds: "Halogen" includes chloro, bromo, fluoro and iodo, and is preferably chloro or bromo and most preferably chloro.

The aliphatic "alkyl" groups may be straight or branched chains containing the specified number of carbon atoms.

The term "pharmaceutically acceptable salt" as used herein is intended to include the nontoxic acid addition salts with inorganic or organic acids, e.g. salts with acids such as hydrochloric, phosphoric, sulfuric, maleic, acetic, citric, succinic, benzoic, fumaric, mandelic, ptoluenesulfonic, methanesulfonic, ascorbic, lactic, gluconic, trifluoracetic, WO 98/23621 PCT/US97/21785 22 hydroiodic, hydrobromic, and the like. Some of the compounds of the present invention have an acidic hydrogen and can, therefore, be converted with bases in a conventional manner into pharmaceutically acceptable salts. Such salts, e.g. ammonium, alkali metal salts, particularly sodium or potassium, alkaline earth metal salts, particularly calcium or magnesium, and salts with suitable organic bases such as lower alkylamines (methylamine, ethylamine, cyclohexylamine, and the like) or with substituted lower alkylamines hydroxyl-substituted alkylamines such as diethanolamine, triethanolamine or tris-(hydroxymethyl)aminomethane), or with bases such as piperidine or morpholine, are also intended to be encompassed by the term "pharmaceutically acceptable salt".

Compounds of the present invention in the form of acid addition salts may be written as x Y H S N x S

CH

CH2-S.Q

COOR'

where X represents an acid anion and R 1 is hydrogen or a carboxylprotecting group. The counter anion X E may be selected so as to provide pharmaceutically acceptable salts for therapeutic administration.

WO 98/23621 PCT/US97/21785 23 The carboxyl-protecting group R' is intended to include readily removable ester groups which have been conventionally employed to block a carboxyl group during the reaction steps used to prepare the compounds of the present invention and which can be removed by methods which do not result in any appreciable destruction of the remaining portion of the molecule, e.g. by chemical or enzymatic hydrolysis, treatment with chemical reducing agents under mild conditions, irradiation with ultraviolet light or catalytic hydrogenation.

Examples of such protecting groups include benzhydryl, p-nitrobenzyl, 2naphthylmethyl, allyl, benzyl, p-methoxybenzyl, trichloroethyl, silyl such as trimethylsilyl, phenacyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl and

C

1

-C

6 alkyl such as methyl, ethyl or t-butyl. Included within such protecting groups are those which are hydrolyzed under physiological conditions such as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl, a-acetoxyethyl, a-pivaloyloxyethyl and methoxymethyl.

Compounds of the present invention with such physiologically hydrolyzable carboxyl protecting groups are also referred to herein as "prodrugs". Compounds of the present invention where R 1 is a physiologically removable protecting group are useful directly as antibacterial agents. Compounds where an R 1 protecting group is not physiologically removable are useful intermediates which can be easily converted to the bioactive form by conventional deblocking procedures well-known to those skilled in the art.

WO 98/23621 PCTIS97/21785 24 Compounds of the present invention wherein a hydroxyl substituent is esterified with a group hydrolyzable under physiological conditions are also included within the scope of the term "prodrug" as used herein. Such hydroxyl-protecting groups may be employed, for example, to increase the solubility of a cephem derivative. Illustrative of suitable ester "prodrugs" of this type are compounds wherein one or more hydroxy substituent groups are converted to sulfate (-OSOH) or phosphate (-OP0 3

H

2 groups.

The compounds of the present invention can be made by conventional methods. A suitable procedure is summarized by the following reaction scheme: WO 98/23621 WO 9823621PCT/US97/21785 i: y

VI

H2N s

CO

2

R

V

IV deprotection x y C0 2

R

IV

x y

HO

2

IV'

N R

III

To elaborate on the above process, intermediates of type VI are first prepared, for example, analogous to the process illustrated below: WO 98/23621 PCT/US9721785 26 Cl Cl C CO 2 t-Bu Cl Bu 3 N, DMF, 80 0 C Cl I Ph 3 P, Pd(OAc) 2

CO

2 t-Bu

S~CO

2

CH

3 NaS CO 2

CH

3 Cl S NaOH DMF Cl

CO

2 t-Bu S

CO

2

H

Cl Cl

CO

2 t-Bu Acid intermediate VI is then coupled with a suitable cephem intermediate having a 3-substituent leaving group. For example, the leaving group may be acetoxy or halogen. In the preferred embodiment illustrated by the reaction scheme, the cephem intermediate is the 3chloromethyl cephem V, but other suitable cephem intermediates with equivalent leaving groups at the 3-position could also be employed. The cephem intermediate V may be acylated with VI or a reactive derivative thereof by conventional acylation procedures well-known in the cephalosporin art to give N-acylated intermediate IV. In addition to using the free arylthioacetic acid, e.g. with a suitable condensing agent such as WO 98/23621 PCT/US97/21785 27 dicyclohexylcarbodiimide, acylating agent VI may also be employed in the form of equivalent acylating derivatives such as an acid anhydride, mixed anhydride, activated ester, or acid halide. The cephem intermediate preferably has the carboxyl group protected by a conventional carboxylprotecting group which can be readily removed. Examples of such protecting groups are discussed above and include benzyl, 4-nitrobenzyl, 1,1 dimethylethyl, 4-methoxybenzyl, diphenylmethyl, allyl, and the like.

Other examples of suitable protecting groups are disclosed in Protective Groups in Organic Synthesis, 2nd Ed., Theodora W. Greene (John Wiley Sons, 1991), Chapter 5. In one embodiment, intermediate V may be acylated with acid VI in the presence of dicyclohexylcarbodiimide and in an inert solvent such as tetrahydrofuran or dichloromethane. The reaction temperature is typically between -20 °C and 50 Upon completion of the reaction, insoluble material is removed by filtration, the filtrate is concentrated, and the residue is treated with a relatively non-polar solvent such as diethyl ether or ethyl acetate resulting in precipitation of the desired product. Alternatively, acid VI may be converted to the corresponding acid chloride, for example by treatment with thionyl chloride with or without a solvent such as dichloromethane, followed by coupling with cephem amine Y in the presence of a base such as triethylamine or N-methylmorpholine to give intermediate IV.

Cephem IV is typically isolated after aqueous work-up and evaporation of volatile solvents followed by trituration of the compound with a relatively non-polar solvent such as diethyl ether or ethyl acetate. This WO 98/23621 PCT/US97/21785 28 intermediate may be used in the next reaction step as the X chloride derivative, or can be converted to the X bromide or X iodide derivative by treatment with the appropriate metal halide in a solvent such as acetone.

To prepare the quaternary cephem I, intermediate IV is deprotected under acidic conditions, followed by reaction of the resulting intermediate IV' with a thiopyridone derivative m. For example, when R is diphenylmethyl or 4-methoxybenzyl, cephem acid IV' is obtained upon treatment of IV with trifluoroacetic acid neat or in an inert solvent such as methylene chloride. A reagent such as anisole may also be employed to scavenge the liberated ester protecting group. The reaction temperature is usually at or below room temperature. The deprotection may also be carried out by treatment with other protic acids such as hydrochloric acid in a solvent such as methanol. The final product is typically isolated by precipitation or crystallization. Reaction of IV' with a thiopyridone derivative m in a solvent such as dimethylformamide, dimethyl sulfoxide, ethanol, methanol, or other appropriate solvents at a temperature between -20 "C and 100 "C affords target quaternary cephem I.

The final product is isolated as described above. Thiopyridones II are typically prepared according to a method analogous to that described in T.

Takahashi et al., European Patent Application No. 209751 and in I.E. El- Kholy et al., J. Heterocyclic Chem. Vol. 11, p. 487 (1974). This procedure entails reaction of 4-thiopyrone (European Patent No. 209751) with an WO 98/23621 PCTIUS97/21785 29 appropriate primary amine in a solvent such as aqueous methanol or ethanol at a temperature ranging between 0 'C and 78 0 C. The primary amine may be in the form of a zwitterion in examples where there is a free acid group present in the molecule. In these cases, a base such as sodium hydroxide, sodium bicarbonate, or pyridine is added to form the free amine in situ. The product may be isolated as its sodium salt by evaporation of volatile solvents, followed by trituration with a solvent such as diethyl ether or ethyl acetate. Alternatively, the reaction mixture may be acidified and extracted with an organic solvent to afford the product as the free carboxylic acid. If the carboxylate group is protected as an ester, the amine may be free or present as an acid salt. In the latter case, a base such as sodium hydroxide, sodium bicarbonate, or pyridine is added to form the free amine in situ. The product is typically isolated by precipitation or by reversed phase column chromatography following removal of volatile solvents.

Other representative intermediate VI groups may be prepared as described in the preparation of starting materials section below.

It will be understood that where the substituent groups used in the above reactions contain certain reaction-sensitive functional groups such as amino or carboxylate groups which might result in undesirable sidereactions, such groups may be protected by conventional protecting groups known to those skilled in the art. For example, thiopyridone WO 98/23621 PCT/US97/21785 intermediates of formula III may have an amine functional group protected as the t-butyloxycarbamate. Suitable protecting groups and methods for their removal are illustrated, for example, in Protective Groups in Organic Synthesis, 2nd Ed., Theodora W. Greene (John Wiley Sons, 1991). It is intended that such "protected" intermediates and endproducts are included within the scope of the present disclosure and claims.

The desired end-product of formula I may be recovered either as the zwitterion or in the form of a pharmaceutically acceptable acid addition salt, e.g. by addition of the appropriate acid such as HC1, HI or methanesulfonic acid to the zwitterion. Compounds of formula I where

R

1 is hydrogen or an anionic charge, or a pharmaceutically acceptable salt thereof, may be converted by conventional procedures to a corresponding compound where R 1 is a physiologically hydrolyzable ester group.

It will be appreciated that certain products within the scope of formula I may have a C-3 substituent group which can result in formation of optical isomers. It is intended that the present invention include within its scope all such optical isomers as well as epimeric mixtures thereof, i.e. R- or S- or racemic forms.

The novel cephalosporin derivatives of general formula I (including the IA compounds) wherein R 1 is hydrogen, an anionic charge WO 98/23621 PCTIUJS97/21785 31 or a physiologically hydrolyzable carboxyl-protecting group, or the pharmaceutically acceptable salts or prodrugs thereof, are potent antibiotics active against many gram-positive bacteria. While they may be used, for example, as animal feed additives for promotion of growth, as preservatives for food, as bactericides in industrial applications, for example in water-based paint and in the white water of paper mills to inhibit the growth of harmful bacteria, and as disinfectants for destroying or inhibiting the growth of harmful bacteria on medical and dental equipment, they are especially useful in the treatment of infectious disease in humans and other animals caused by the gram-positive bacteria sensitive to the new derivatives. Because of their excellent activity against MRSA organisms, they are particularly useful in the treatment of infections resulting from such bacteria.

The pharmaceutically active compounds of this invention may be used alone or formulated as pharmaceutical compositions comprising, in addition to the active cephem ingredient, a pharmaceutically acceptable carrier or diluent. The compounds may be administered by a variety of means, for example, orally, topically or parenterally (intravenous or intramuscular injection). The pharmaceutical compositions may be in solid form such as capsules, tablets, powders, etc. or in liquid form such as solutions, suspensions or emulsions. Compositions for injection, the preferred route of delivery, may be prepared in unit dose form in ampules or in multidose containers and may contain additives such as suspending, WO 98/23621 PCT/US97/21785 32 stabilizing and dispersing agents. The compositions may be in ready-touse form or in powder form for reconstitution at the time of delivery with a suitable vehicle such as sterile water.

The dosage to be administered depends, to a large extent, on the particular compound being used, the particular composition formulated, the route of administration, the nature and condition of the host and the particular situs and organism being treated. Selection of the particular preferred dosage and route of application, then, is left to the discretion of the physician or veterinarian. In general, however, the compounds may be administered parenterally or orally to mammalian hosts in an amount of from about 50 mg/day to about 20 g/day. Administration is generally carried out in divided doses, three to four times a day, analogous to dosing with a cephalosporin such as cefotaxime.

IN VITRO ACTIVITY Samples of the compounds prepared below in Examples 1 22 after solution in water and dilution with Nutrient Broth were found to exhibit the following Minimum Inhibitory Concentrations (MIC) values versus the indicated microorganisms as determined by tube dilution. The MICs were determined using a broth micro dilution assay in accordance with that recommended by the National Committee for Clinical Laboratory Standards (NCCLS). Mueller-Hinton medium was used except for WO 98/23621 PCT/US97/21785 33 Streptococci which was tested in Todd Hewitt broth. The final bacterial inoculate contained approximately 5 x 10 5 cfu/ml and the plates were incubated at 35 0 C for 18 hours in ambient air (Streptococci in 5% C02).

The MIC was defined as the lowest drug concentration that prevented visible growth.

Microorganism MIC value in ug/ml S. aureus methicillin resistant A27223 <8 S. pneumoniae A9585 <2 S. pyogenes A9604 <2 E. faecalis A20688 <8 S. aureus A9537, penicillinase negative <1 S. aureus A15090, penicillinase positive <1 S. epidermidis A24548 <1 S. epidermidis A25783, methicillin resistant <2 S. hemolyticus A21638 <8 WO 98/23621 PCT/US97/21785 34 IN VIVO ACTIVITY The in vivo therapeutic efficacy of the compounds prepared in Examples 1 22 below after intramuscular injection to mice experimentally infected with the representative MRSA strain A27223 was also measured.

The determination of the effectiveness of antimicrobial agents in Staphylococcus aureus systemic infection in mice Organisms: The test organism, MRSA strain A27223 used to generate systemic infection in mice, is grown on two large Brain Heart Infusion Agar plates. On each plate, 0.5 ml of frozen stock culture is plated out.

Plates are then incubated for 18 hours at 30 0 C. The next day each plate is washed with 20 ml of Brain Heart Infusion Broth and then pooled together. A microscopic direct count of microorganism is done using a 1:1000 dilution of plate wash. After a direct count is obtained, the number of organisms per milliliter is calculated. The count is adjusted to the desired amount of inoculum by diluting in 4% hog mucin. The desired challenge (amount of organisms given to mice) is 2.4 x 108 ml/mouse for MRSA strain A27223. The mice are infected intraperitoneally with 0.5 ml of challenge. Ten non-treated infected mice are used as controls.

WO 98/23621 PCT/US97/21785 Mice: Mice used are male ICR mice. The average weight of the animals is from 20 to 26 grams.

Drug preparation and treatment: Compounds are tested at 4 dose levels, (25, 6.25, 1.56, and 0.39 mg/kg) and prepared in 5% cremophor, unless otherwise specified. Vancomycin is used as the control compound, and is dosed at 6.25, 1.56, 0.39, and 0.098 mg/kg. It is prepared in 0.1M phosphate buffer. There are five infected mice per dose level, and they are treated with 0.2 ml of the test compound, preferably by intramuscular injection.

Treatment begins 15 minutes and 2 hours post-infection.

Test duration: A PD50 (the dose of drug given which protects 50% of mice from mortality) experiment runs for 5 days. During this time, mortality of mice are checked every day and deaths are recorded. The cumulative mortality at each dose level is used to calculate a PD50 value for each compound. Surviving mice are sacrificed at the end of day 5 by C02 inhalation.

Calculation: Actual calculation of PD50 is performed with a computer program using the Spearman-Karber procedure.

Results: The in vivo efficacy, expressed as the PD50 value, ranged from about 0.8 to 22.0 mg/kg (for certain compounds, more than one test was WO 98/23621 PCT/US97/21785 36 carried out; the indicated range is for at least one test result when multiple tests were done).

ILLUSTRATIVE EXAMPLES The following examples illustrate the invention, but are not intended as a limitation thereof. The abbreviations used in the examples are conventional abbreviations well-known to those skilled in the art.

Some of the abbreviations used are as follows: h hour(s) mol mole(s) mmol mmole(s) g gram(s) THF tetrahydrofuran L liter(s) mL milliliter(s) Et 2 0 diethyl ether EtOAc ethyl acetate BOC butoxycarbonyl DCC dicyclohexylcarbodiimide DPM diphenylmethyl Ph 3 P triphenylphosphine t-Bu tertiary-butyl EtOH ethanol WO 98/23621 PCT/US97/21785 37 Bu 3 N tributylamine MeOH methanol DMF dimethylformamide DABCO 1,4-diazabicyclo[2.2.2]octane TFA trifluoroacetic acid DMS dimethylsulfide m-CPBA meta-chloroperoxybenzoic acid TFAA trifluoroacetic anhydride TEA triethylamine In the following examples, all temperatures are given in degrees Centigrade. Proton nuclear magnetic resonance 1 H NMR) spectra were recorded on a Bruker AM-300 or a Varian Gemini 300 spectrometer. All spectra were determined in CDCl 3 DMSO-d 6 CD30D, or D 2 0 unless otherwise indicated. Chemical shifts are reported in 8 units relative to tetramethylsilane (TMS) or a reference solvent peak and interproton coupling constants are reported in Hertz Splitting patterns are designated as follows: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br, broad peak; dd, doublet of doublets; dt, doublet of triplets; and app d, apparent doublet, etc. Mass spectra were recorded on a Kratos or a Finnegan 4500 instrument utilizing direct chemical ionization (DCI, isobutene), fast atom bombardment (FAB), or electron ion spray

(ESI).

WO 98/23621 PCT/US97/21785 38 Analytical thin-layer chromatography (TLC) was carried out on precoated silica gel plates (60F-254) and visualized using UV light, iodine vapors, and/or staining by heating with methanolic phosphomolybdic acid. Column chromatography, also referred to as flash chromatography, was performed in a glass column using finely divided silica gel at pressures somewhat above atmospheric pressure with the indicated solvents. Reversed-phase analytical thin-layer chromatography was carried out on precoated reverse phase plates and visualized using UV light or iodine vapors. Reversed-phase column chromatography was performed in a glass column using Baker Octadecyl (C 18 40 mm.

WO 98/23621 WO 9823621PCTIUS97/21785 Preparation of Starting Materials scheme 2 s -lCO 2

H

00 2 t-Bu

H

2 Pt0 2 S,,C0 2

H

CI

2 00 2 t-Bu scheme 3 S0N 02 Me O l

CI

0No 4 C0 2 H DCC, THF 5

H

2 N ,.ICO2t-BU S ~.CO 2 Me HN 0 L- CO 2 t-Bu scheme 4 CI CI CI 03~ Jones Cil Cl DMS

CC

-l CCHO C02H~ 7 CO 2 t-Bu 8

H

2 S0 4

CI

C0 2 t-BU NaS ll/C02Me

DMF

CO

2 t-Bu 11 WO 98/23621 WO 9823621PCTIUJS97/2 1785 scheme ile S ,,CO2Me H2N CO2t-Bu TFA CI I ul

DCC

C0 2

H

12 S,.,,CO2Me Clf~ HN 0 13 CO2t-Bu

CO

2 t-Bu 11 scheme 6 Ci

CI

Ci Q Br,,,C02t-Bu Cl. m-CPBA Cl Ci Ileq.

14 SH 15 S, ,CO2t-Bu m-CPBA 2 eq.

10i NaS N/ C02Me C

DMF

\CO

2 t-Bu 0 "S C 2 t-Bu 17 INal

TFAA

5 ,C 2 Me

CI\

S

C

2 t-Bu NaS I/CO2Me

DMF

0 Pp 2 t-Bu scheme 7

CI

IlCN NaS ,Co 2 Me Ci~t H N C o t- I l0 I l Et 3 N DMF S0 2 CI HN1~ tB 21 VN\ 0tB S CO02Me 22 23 WO 98/23621 PCT/US97/21785 41 scheme 8 S O OH 12 OH Ox

N

SA

N

C gSO C O,4 Me 2 NC(S)CI CA 220C Cl~ A 220oC Cl CH 2 C1 2 Cl Cl "L Cl 24 1 25 26 SH S CO 2 Me S CO2Me KOH CIl TEA Cl C 0_Io 1 n-Bu 3 Sn o CO2t-Bu C C BrCH 2 COMe 'Cl 29 Cl 27 28 I n-Bu 3 Sn CO t-Bu n-Bu3Sn CO2t-Bu S CO 2 Me 31

CI

CO

2 t-Bu 32 A. Synthesis of acid 2 Acid 1 (2.00 0.006 mol) is dissolved in 50 mL EtOH, and PtO2 (1.00 0.004 mol) is added. The mixture is hydrogenated at 20 psi for 18 hours.

1 H-NMR analysis indicates that the reaction has only proceeded to conversion. More PtO2 (0.400 0.002 mol) is added, and hydrogenation at 20 psi is continued another 6 hours. At this time the reaction is only complete. The solids are filtered, and the filtrate is treated with fresh Pt02 (1.00 0.004 mol) and hydrogenated at 20 psi for another 20 hours.

The reaction still shows some starting olefin. At this point the mixture is filtered, and the filtrate is concentrated. The residue is chromatographed on silica using CH2C12 as eluant, followed by a gradient elution with WO 98/23621 PCT/US97/21785 42 methanol/CH2Cl2 (up to 15% MeOH). Acid 2 is obtained (0.750 0.002 mol; 33% yield) as a white solid.

1 H-NMR (300 MHz, CDC13): 8 7.40 1H, ArH), 7.30 1H, ArH), 3.72 (s, 2H, SCH2), 2.96 2H, J 8 Hz, ArCH2), 2.54 2H, J 8 Hz, CH2CO2R), 1.43 9H, C(CH3)3).

B. Synthesis of acid 4 Diester 3 (7.00 0.019 mol) is suspended in 40 mL CH2C12. Anisole (1 mL) is added, followed by 15 mL of trifluoroacetic acid. The mixture is stirred for 1 hour at room temperature. The solvents are concentrated to mL, and excess ether is added to precipitate a white solid. The solid is collected, washed with ether, and dried under vacuum to yield acid 4 (4.85 0.015 mol; 79% yield).

1 H-NMR (300 MHz, DMSO-d6): 8 8.08 1H, ArH), 7.71 1H, J 16 Hz, ArCH=C), 7.43 1H, ArH), 6.69 1H, J 16 Hz, C=CHCO2), 4.19 2H, SCH2), 3.66 3H, OCH3).

WO 98/23621 PCT/US97/21785 43 C. Synthesis of hydroxysuccinimide ester Acid 4 (8.75 0.027 mol) is suspended in 55 mL THF under an atmosphere of nitrogen. Dicyclohexylcarbodiimide (1M in CH2C12, 28.7 mL, 0.029 mol) is added, followed by N-hydroxysuccinimide (3.14 0.027 mol). The reaction is allowed to stir for 3 hours at room temperature.

The mixture is diluted with -30 mL acetone, filtered to remove dicyclohexylurea, and concentrated to -25 mL. A solid forms which is filtered off, and the filtrate is evaporated to dryness. Crude 5 is obtained as a white solid (12.3 which is of sufficient purity for use in subsequent reactions.

1 H-NMR (300 MHz, DMSO-d6): 8 8.27 1H, ArH), 8.01 1H, J 16 Hz, ArCH=C), 7.50 1H, ArH), 7.17 1H, J 16 Hz, C=CHCO2), 4.23 2H, SCH2), 3.68 3H, OCH3), 2.84 4H, CH2CH2).

D. Synthesis of amide 6 tert-Butylglycine hydrochloride (5.19 0.031 mol) is suspended under a nitrogen atmosphere in 60 mL dry DMF. N-Methylmorpholine (3.90 mL, 0.036 mol) is added, and then the mixture is cooled to 0°C.

Crude hydroxysuccinimide ester 5 (12.3 is added, and the mixture allowed to stir for 10 minutes at 0°C. The cooling bath is removed and the reaction is stirred for 1 hour. The mixture is concentrated, and the residue dissolved in ethyl acetate and placed in a separatory funnel. The solution is washed with 0.4 N aqueous HC1, 0.1 N aqueous NaHCO3, WO 98/23621 PCT/US97/21785 44 water and then brine. The organic phase is dried (MgSO4) and evaporated to afford clean amide 6 as a light yellow solid (10.4 0.024 mol; 89% yield from acid 4).

1 H-NMR (300 MHz, DMSO-d6): 8 8.42 1H, J 8 Hz, NH), 7.81 1H, ArH), 7.59 1H, J 16 Hz, ArCH=C), 7.47 1H, ArH), 6.80 1H, J 16 Hz, C=CHCONH), 4.20 2H, SCH2), 3.85 2H, J 8 Hz, NCH2CO2), 3.67 3H, OCH3), 1.41 9H, C(CH3)3).

E. Synthesis of 2.4,5-trichlorobenzaldehyde 8 Ester 7 (1.03 3.35 mmol) is dissolved in 200 mL CH2C12 and 100 mL methanol. The mixture is cooled to -78 0 C, and ozone is bubbled through the solution until it turns blue. The mixture is purged with oxygen, and then 1.3 mL of methyl sulfide is added. The mixture is allowed to warm to room temperature, and the solvents are evaporated.

The crude residue is partitioned between ether and water. The ether layer is washed with water and brine, and then dried (MgSO4). Concentration of the ether layer, followed by chromatography on silica using CH2Cl2/hexane as eluant, affords 8 (0.55 2.63 mmol; 79% yield) as a white solid.

1 H-NMR (300 MHz, CDC13): 8 10.34 1H, CHO), 7.98 1H, ArH), 7.58 (s, 1H, ArH).

WO 98/23621 P~rUS97/21785 F. Synthesis of 2,4,5-trichlorobenzoic acid 9 2,4,5-Trichlorobenzaldehyde (0.275 1.31 mmol) is dissolved in 8 mL acetone. A slight excess of Jones Reagent is added, and the mixture stirred at room temperature for 1 hour. Methanol mL) is added, and after 5 minutes the mixture is partitioned between methylene chloride and water. The aqueous phase is extracted with chloroform and the combined organic extracts are washed with water, then brine. The organic phase is dried (MgSO4), and concentrated to afford pure 2,4,5trichlorobenzoic acid 9 (0.285 1.26 mmol; 96% yield) as a white solid.

1 H-NMR (300 MHz, CDCI3): 8 8.01 1H, ArH), 7.61 1H, ArH).

G. Synthesis of 2,4,5-trichlorobenzoic acid, tert-butyl ester Acid 9 (2.93 13.0 mmol) placed in a Parr hydrogenation bottle (under an atmosphere of nitrogen) is dissolved in 55 mL of dioxane. The bottle is cooled to -78 0 C, and 5 mL conc. sulfuric acid is added cautiously, followed by -50 mL liquid isobutylene (cooled to -78 0 The bottle is sealed and agitated on a Parr shaker overnight (-19.5 hours). The sealed bottled is vented, and the solution slowly added to a separatory funnel containing half-saturated aqueous NaHCO3 and ether. The aqueous layer is extracted with ether, and the combined organic phase was dried (MgSO4) and concentrated. Chromatography on silica using hexane, followed by 25% CH2C12/hexane, affords ester 10 as a pale yellow oil (2.56 WO 98/23621 PCT/US97/21785 46 9.10 mmol; 70% yield) which solidifies overnight in the refrigerator to an off-white solid.

1 H-NMR (300 MHz, CDC13): 8 7.81 1H, ArH), 7.53 1H, ArH), 1.58 (s, 9H, C(CH3)3).

H. Synthesis of diester 11 Using the method described below for the synthesis of diester 3, ester 10 (2.5 11.3 mmol) is converted to diester 11 (3.20 9.12 mmol; 81% yield). Diester 11 was obtained as a white solid by chromatography on silica gel using 80% methylene chloride/hexane.

1 H-NMR (300 MHz, CDC13): 6 7.75 1H, ArH), 7.30 1H, ArH), 3.75 (s, 3H, OCH3), 3.72 2H, SCH2), 1.58 9H, C(CH3)3).

I. Synthesis of acid 12 Diester 11 (1.60 4.56 mmol) is dissolved in 5 mL methylene chloride. Trifluoroacetic acid (2 mL) is added, and the mixture stirred for 4 hours at room temperature. The solvents are evaporated to provide 12 (1.24 4.20 mmol; 92% yield) as a tan solid of sufficient purity for use in the next reaction.

WO 98/23621 PCTfUS97/21785 47 1 H-NMR (300 MHz, DMSO-d6): 8 7.87 1H, ArH), 7.46 1H, ArH), 4.23 2H, SCH2), 3.68 3H, OCH3).

J. Synthesis of amide 13 Acid 12 (1.24 4.20 mmol) is dissolved in 14 mL dry THF.

Dicyclohexylcarbodiimide (0.866 4.20 mmol) is added, followed by tertbutyl glycine (0.550 4.20 mmol), and the mixture stirred at room temperature for 2 hours. Ether is added to the flask, and the solids removed by filtration. The filtrate is evaporated to yield 1.96 g. of crude material. Chromatography on silica using 80% CH2C12/hexane, followed by a second chromatography using 20% ethyl acetate/hexane affords amide 13 (0.844 2.07 mmol; 49% yield) as a white solid.

1 H-NMR (300 MHz, CDC13): 6 7.78 1H, ArH), 7.31 1H, ArH), 4.12 (d, 2H, J 6 Hz, NCH2CO 2 3.77 3H, OCH3), 3.72 2H, SCH2), 1.48 9H, C(CH3)3).

K. Synthesis of ester 2,4,5-Trichlorothiophenol (35.0 0.166 mol) is dissolved in 500 mL methylene chloride and cooled to 0°C. Triethylamine (22.0 0.217 mol) is added, followed by a solution tert-butyl bromoacetate (35.1 0.180 mol) in 100 mL methylene chloride (over 5 minutes). After stirring for minutes at 0°C, the ice-bath is removed, and stirring continued for WO 98/23621 PCTIUS97/21785 48 another hour. The mixture is placed in a separatory funnel and washed with water 10% aqueous H3P04, and then brine. The organic phase is dried (MgSO4), and evaporated to afford a white solid, which is washed with hexane. Ester 15 (51 0.156 mol; 94%) as obtained is of suitable purity for subsequent reactions.

1 H-NMR (300 MHz, CDC13): 8 7.47 1H, ArH), 7.45 1H, ArH), 3.66 (s, 2H, SCH2), 1.43 9H, C(CH3)3).

L. Synthesis of sulfoxide 16 Ester 15 (50.0 0.153 mol) is dissolved in 500 mL chloroform and cooled to 0°C. m-Chloroperoxybenzoic acid (50-60% from Aldrich, 48.0 g., 0.140-0.168 mol) is added in small portions over 30 minutes. The ice bath is removed and stirring continued for 2.5 hours at room temperature.

The solids were removed by filtration, and the filtrate washed with dilute aqueous NaHSO3, 5% aqueous Na2CO3, saturated aqueous NaHCO3, and then brine. The organic phase is dried (MgSO4), and evaporated. The crude material is chromatographed twice on silica gel using methylene chloride and then 3% methanol/methylene chloride to afford sulfoxide 16 (35.0 0.102 mol; 67% yield) as a white solid.

WO 98/23621 PCT/US97/21785 49 1 H-NMR (300 MHz, CDCI3): 8 7.95 1H, ArH), 7.53 1H, ArH), 4.05 (d, 1H, J 14 Hz, CH2S(O)), 3.70 1H, J 14 Hz, CH2S(O)), 1.43 9H, C(CH3)3).

M. Synthesis of diester 17 Using the procedure described below for the synthesis of diester 3, sulfoxide 16 (35.0 105 mmol) is converted to diester 17 (32.5 78.7 mmol; 75% yield). The compound is isolated as a white solid after chromatography on silica using methylene chloride then 3% methanol/methylene chloride.

1 H-NMR (300 MHz, CDC13): 8 7.83 1H, ArH), 7.28 1H, ArH), 3.92 (d, 1H, J 14 Hz, CH2S(O)), 3.77 3H, OCH3), 3.71 2H, SCH2), 3.57 1H, J 14 Hz, CH2S(O)).

N. Synthesis of diester Diester 17 (28.0 67.8 mmol) is dissolved in 500 mL acetone.

Sodium iodide (48.7 325 mmol) is added, followed by trifluoroacetic anhydride (40.0 191 mmol) over 5 minutes. After stirring at room temperature for 1 hour, the solvents are evaporated. Methylene chloride is added and evaporated twice. The residue is taken up in methylene chloride and washed with aqueous NaHSO3 solution water, and then brine. The organic phase is dried (MgSO4), and evaporated.

WO 98/23621 PCTfUS97/21785 Chromatography of the residue on silica using methylene chloride affords diester 20 (23.5 59.2 mmol; 87 yield) as a white solid.

1 H-NMR (300 MHz, CDCl3): 8 7.38 1H, ArH), 7.35 1H, ArH), 3.74 (s, 3H, OCH3), 3.66 2H, SCH2), 3.60 2H, SCH2).

O. Synthesis of sulfone 18 Ester 15 (7.00 21.4 mmol) is dissolved in 40 mL chloroform and treated with m-chloroperoxybenzoic acid from Aldrich, 12.0 -42 mmol). After stirring for 1 hour at room temperature, the solids are removed by filtration, and the filtrate is washed with dilute aqueous NaHSO3, 5% aqueous Na2CO3, saturated aqueous NaHCO3, and then brine. The organic phase is dried (MgSO4), and evaporated.

Chromatography of the residue on silica using methylene chloride then 3% methanol/methylene chloride affords sulfone 18 (7.00 19.5 mmol; 93% yield) as a white solid.

1 H-NMR (300 MHz, CDC13): 8 8.17 1H, ArH), 7.67 1H, ArH), 4.32 (s, 2H, SCH2), 1.33 9H, C(CH3)3).

P. Synthesis of diester 19 Using the procedure described below for the synthesis of diester 3, sulfone 18 (7.20 20.0 mmol) is converted to diester 19 (8.05 18.8 WO 98/23621 PCT/US97/21785 51 mmol; 94% yield). The compound is isolated as a white solid after chromatography on silica using methylene chloride then 3% methanol/methylene chloride.

1 H-NMR (300 MHz, CDC13): 8 8.05 1H, ArH), 7.40 1H, ArH), 4.28 (s, 2H, SO2CH2), 3.78 3H, OCH3), 3.77 2H, SCH2).

Q. Synthesis of sulfonamide 22 tert-Butyl glycine (2.62 20.0 mmol) and triethylamine (2.50 25.0 mmol) are dissolved in 20 mL chloroform under a nitrogen atmosphere.

The solution is cooled with an ice-bath, and 2,4,5trichlorobenzenesulfonyl chloride (5.60 20.0 mmol) dissolved in 30 mL chloroform is added over 5 minutes. The cooling bath is removed and the mixture allowed to stir at room temperature for 1 hour. The solution is washed with 10% aqueous H3P04, water, and then brine. The organic phase is dried (MgSO4), and evaporated to afford clean sulfonamide 22 (7.00 18.8 mmol; 94% yield) as a white solid.

1 H-NMR (300 MHz, CDC13): 8 8.10 1H, ArH), 7.63 1H, ArH), 3.72 (s, 2H, NCH2CO2), 1.33 9H, C(CH3)3).

R. Synthesis of diester 23 Using the procedure described above, sulfonamide 22 (3.70 10.0 mmol) is converted to diester 23 (2.37 7.40 mmol; 74% yield). The WO 98/23621 PCT/US97/21785 52 compound is isolated as a white solid after chromatography on silica using methylene chloride then 5% methanol/methylene chloride.

1 H-NMR (300 MHz, CDC1 3 8 7.96 1H, ArH), 7.36 1H, ArH), 5.65 (t, 1H, J 5 Hz, NH), 3.85 3H, OCH3), 3.78 2H, SCH2), 3.71 2H, J Hz, NCH2CO 2 1.34 9H, C(CH 3 3 S. Synthesis of 2.5-dichloro-4-iodophenol 24 2 ,5-Dichlorophenol (20.4 0.125 mol) is placed in a 1L round bottom flask equipped with a large egg-shaped stir bar and is dissolved in 307 mL CH2C12. With rapid stirring, iodine (46.6 0.183 mol) is added, followed by silver sulfate (42.3 0.136 mol). The purple solution is stirred 1 day, at which point NMR analysis of an aliquot indicates the reaction is complete.

The reaction is diluted with CH2C12 (-200 ml) and filtered through a fritted Buchner funnel to remove silver salts. The salts are washed with additional CH2C12 (-100 mL). The organic filtrate is transferred to a separatory funnel and is washed first with a solution of sodium thiosulfate (-40 g. in -200 mL water; this removes excess 12), and then brine. The organic phase is dried (MgSO4), and evaporated to give 24 (34.59 0.108 mol; 86% yield) as a pale pink/yellow solid. The material is sufficiently pure to carry forward.

WO 98/23621 PCT/US97/21785 53 1 H-NMR (300 MHz, CDC13): 8 7.75 1H, ArH), 7.14 1H, ArH), 5.62 (br, 1H, OH).

T. Synthesis of thiocarbamate Iodophenol 24 (34.59 0.108 mol) is placed into a 500 mL round bottom flask equipped with septa, N2 inlet, and a stir bar. The phenol is then dissolved in 130 mL DMF. DABCO (24.2 0.216 mol) is added followed by dimethylthiocarbamyl chloride (21.6 0.175 mol). The mixture is stirred at room temperature for 1 hour, then diluted with EtOAc (-400 mL) and poured into a separatory funnel containing 300 mL of ice-water. The phases are separated, and the aqueous extracted twice with 200 mL of EtOAc. The combined organic extracts are washed twice with water 100 mL), and then brine. The organic phase is dried (MgSO4), and evaporated to afford 25 as a dark oil. This material is dissolved in CH2C12 and dried again (MgSO4). After evaporation a yellow solid (-35 is obtained. The compound was of sufficient purity to be used in the next reaction.

1 H-NMR (300 MHz, CDC13): 8 7.90 1H, ArH), 7.24 1H, ArH), 3.46 (s, 3H, CH3), 3.37 3H, CH3).

WO 98/23621 PCT/US97/21785 54 U. Synthesis of 26 The crude material obtained above (-35 was heated neat under N2 at 220 0 C for two hours. After cooling, the material was dissolved in CH2C1 2 and filtered through a plug of silica gel. The fractions containing the product are evaporated to afford 30.2 g. of a brown solid. This material was chromatographed on silica (in portions) using a gradient elution starting with 1:1 CH2Cl2/hexane (material dissolved in a minimum amount of CH2C12 for column loading), and then -70% CH2Cl2/hexane.

Compound 26 is obtained as a yellow crystalline solid (13.0 36.0 mmol; 33% yield from 2 1 H-NMR (300 MHz, CDC13): 8 7.95 1H, ArH), 7.62 1H, ArH), 3.10 (br s, 3H, CH3), 3.00 (br s, 3H, CH3).

V. Synthesis of 2 ,5-dichloro-4-iodothiophenol 27 Carbamate 26 (9.80 0.026 mol) is dissolved in 40 mL EtOH and treated with 30 mL 3N aqueous KOH. The mixture is heated to reflux with stirring under nitrogen for 3 hours. The solution is allowed to cool and is then acidified with 3 N HCI until pH The mixture is extracted with CH2C12 (thrice), and the combined organic phase washed with water and then brine. The extracts are dried (MgSO4) and evaporated. The crude material is chromatographed on silica using 1:1 CH2C12/hexane.

Thiol 27 (6.43 0.021 mol; 81% yield) is obtained as a white solid.

WO 98/23621 PCT~US97/21785 1 H-NMR (300 MHz, CDC13): 8 7.83 1H, ArH), 7.56 1H, ArH).

W. Synthesis of ester 28 Thiol 27 (6.43 0.021 mol) is dissolved in 50 mL CH2C12 and triethylamine (2.52 0.025 mol) is added. Methyl bromoacetate (3.82 g., 0.025 mol) is then added over 5 minutes. The resultant mixture is stirred at room temperature for 1.5 hours, at which time 1 H-NMR analysis indicated the reaction was complete. The mixture was diluted with CH2C12 (-200 mL) and was washed with water, 1N HC1, water, and then brine. The organic layer was dried (MgSO4) and evaporated. The crude material is chromatographed on silica using 70% CH2C12/hexane. Ester 28 is obtained as a white solid (7.20 0.019 mol; 90% yield).

1 H-NMR (300 MHz, CDC13): 8 7.82 1H, ArH), 7.42 1H, ArH), 3.75 (s, 3H, OCH3), 3.68 2H, SCH2).

X. Synthesis of stannane 29 Bis(tributyltin) (29.5 50.9 mmol) is dissolved under a nitrogen atmosphere in 70 mL dry THF. The solution is cooled to -20 0 C, and butyllithium (1.6 M in hexane, 31.2 mL, 49.9 mmol) is added dropwise over 20 minutes, maintaining the temperature of the bath at -20 0 C. The solution is then cooled to -50 0 C, and then copper(I) bromide WO QQ1 /21* 1 *n^^m~Tnrronrf- f 56 methylsulfide complex (5.10 24.8 mmol) is added. The mixture is allowed to stir at -40 0 C for 15 minutes, and is then cooled to -78 0 C. Bromofuroic acid tert-butyl ester (4.10 16.6 mmol) dissolved in 15 mL THF is added, and the mixture allowed to stir for 3 hours at -78 0 C. The reaction mixture is poured into 1 L of ether and -300 mL half-saturated aqueous ammonium chloride solution. After stirring for 5 minutes the ether layer is decanted onto another -300 mL of half-saturated aqueous ammonium chloride solution. After 5 minutes the biphasic mixture is separated, and the organic phase is washed with brine, dried (MgSO4) and evaporated. Chromatography on silica using hexane, then methylene chloride/hexane affords stannane 29 (5.05 11.1 mmol; 67% yield) as a clear oil.

1 H-NMR (300 MHz, CDC13): 8 7.04 1H, J 4 Hz, HetArH), 6.56 1H, J 4 Hz, HetArH), 1.59-1.47 3 H, SnBu3), 1.37-1.24 9 H, SnBu3), 1.13- 1.05 6 H, SnBu3), 0.89 9H, J 6 Hz, SnBu3).

Y. Synthesis of diester Stannane 29 (1.50 3.28 mmol) is dissolved in 8 mL dry THF. Aryl iodide 28 (0.928 2.46 mmol) is added, followed by bis(triphenylphosphine)-palladium(II) chloride (0.160 0.228 mmol).

The solution is heated to reflux for 6 hours. The mixture is diluted with mL THF, 4 mL conc. aqueous KF is added, and the mixture is stirred for 20 minutes. Ether is added, and the mixture is then filtered to remove 3 WO 98/23621 PCTIUS97/21785 57 insoluble tin solids. The biphasic filtrate is separated, and the aqueous layer is extracted with ether. The combined organic phases are washed with brine, dried (MgSO4) and evaporated. During evaporation crystals began to form, and when only ~5 mL of liquid remains it is decanted. The solids are washed with hexane and then pumped dry. Diester 30 (0.793 g., 1.91 mmol; 78% yield) is obtained as a white solid.

1 H-NMR (300 MHz, CDC13): 8 7.99 (br s, 1H, ArH), 7.40 (br s, 1H, ArH), 7.19 1H, J 2 Hz, HetArH), 7.13 1H, J 2 Hz, HetArH), 3.75 3H, OCH3), 3.71 2H, SCH2), 1.60 9H, C(CH3)3).

Z. Synthesis of stannane 31 Using the method described above, 5-bromo-2-thiophenecarboxylic acid, tert-butyl ester (4.52 17.2 mmol) is converted to stannane 31 (4.33 9.16 mmol; 53% yield). The compound is isolated as a light yellow oil after chromatography on silica using 25% methylene chloride/hexane.

1 H-NMR (300 MHz, CDC13): 8 7.80 1H, J 3 Hz, HetArH), 7.12 1H, J 3 Hz, HetArH), 1.58 15H, SnBu3), 0.92 9H, J 6 Hz, SnBu3), 0.85- 0.80 3H, SnBu3).

WO 98/23621 PCT/US97/21785 58 AA. Synthesis of diester 32 Using the method described above for the synthesis of diester stannane 31 (1.70 3.60 mmol) and aryl iodide 28 2.73 mmol) are converted to diester 32 (0.920 2.13 mmol; 78% yield as a light yellow solid).

1 H-NMR (300 MHz, CDC13): 5 7.66 1H, J 3 Hz, HetArH), 7.55 1H, ArH), 7.44 1H, ArH), 7.27 1H, J 3 Hz, HetArH), 3.78 3H, OCH3), 3.73 2H, SCH2), 1.58 9H, C(CH3)3).

WO 98/23621 PTU9/18 PCTIUS97/21785 59 Example 1 1- 12 -hydroxy-3-amino-p2rop-l-yll-4-[f(6R)-trans-2-carboxy-8..xoL2.5 dichloro-4-(2-carboxyethenyl)henylthio)acetamidol5thia..1 azabicyclo[4.2.01-oct-2-en-3-lmethylthiolpyridinium inner salt.

scheme I ClI C1 -:-c0 2 t-Bu L~iCI BU 3 N, DMF, 80'C Ph 3 P, Pd(OAc) 2 s ,C02F NaOH CIN cl C1 NaS.,C2G SC0 2 Me C1 DMF NP C0'MU C02t-BuCO 2 t-BU 01 0 V' C02 DPM C1 C0 2

DPM

DCC -o-1

TFA

In, C0 2

H-

IA'

Cl.

HA,, NHBOC C02H

TFA

I NH3+ C02H A. Synthesis of olefin 7 2,4,5-Trichloroiodobenzene (25 81.3 mmol) is dissolved in 80 mL DMF. tert-Butyl acrylate (48 mL, 328 mmol), tributylamine (58 mL, 243 mmol), triphenyiphosphine (4.08 15.5 mmol), and palladium acetate (3.23 14.4 mmol) are added, and the mixture heated to 80 0 C for three WO 98/23621 PCTIUS97/21785 hours. The solvents are evaporated and the residue is partitioned with EtOAc and water. The aqueous phase is extracted with EtOAc, and the combined organic phase is washed with brine, dried (MgSO4), and evaporated. The dark red-brown oil is chromatographed on silica in a fritted Buchner funnel using vacuum filtration, with CH2Cl2/hexane followed by 50% CH2Cl2/hexane as eluant. Acrylate 7 is obtained (18.7 60.8 mmol; 75% yield) as a mauve solid.

1 H-NMR (300 MHz, CDC13): 8 7.82 1H, J 16 Hz, ArCH=C), 7.67 1H, ArH), 7.51 1H, ArH), 6.34 1H, J 16 Hz, C=CHCO2t-Bu), 1.51 9H, C(CH3)3).

B. Synthesis of diester 3 Acrylate 7 (21.23 69 mmol) is dissolved in 131 mL DMF. The sodium salt of methyl mercaptoacetate (17.7 crude: see note below) is added and the mixture stirred at room temperature for one hour. The mixture is partitioned with EtOAc and water. The aqueous phase is extracted with EtOAc, and the combined organic phase is washed with brine, dried (MgSO4), and evaporated. Chromatography on silica in a fritted Buchner funnel (vacuum filtration) using 50% CH2C12/hexane followed by 90% CH2Cl2/hexane as eluant affords diester 3 (19.6 52.0 mmol; 75% yield).

WO 98/23621 PCT/US97/21785 61 1 H-NMR (300 MHz, CDC13): 8 7.86 1H, J 16 Hz, ArCH=C), 7.60 1H, ArH), 7.36 1H, ArH), 6.35 1H, J 16 Hz, C=CHCO2t-Bu), 3.77 3H, OCH3), 3.72 2H, SCH2), 1.51 9H, C(CH3)3).

Note: The sodium salt of methyl mercaptoacetate is best made fresh before use. Approximately 30 mL of methyl mercaptoacetate is dissolved in -250 mL THF. One equivalent of 5 N NaOH is added slowly in pipetfulls, and the mixture allowed to stir for 5 minutes. The solvents are removed in vacuo (including water) and the sticky solid is co-evaporated with ether (-200 mLs) and then dry THF (2 x 200 mLs). The solid is pumped dry for several hours under high vacuum until the flask is no longer cool due to evaporation. The freely mobile white solid is used as obtained. Excess of this reagent (1.5 to 2 equivalents) is generally used.

C. Synthesis of acid 1 Diester 3 (4.40 0.012 mol) is dissolved in 30 mL THF. To this solution is added 13 mL of 1N NaOH, and the mixture is allowed to stir at room temperature for 1.5 hours. At this time 1 H-NMR analysis of an aliquot indicates that the reaction is complete. The THF is removed under vacuum, and the concentrate is diluted with water and extracted with EtOAc. The aqueous layer is then acidified with 1N HC1 to pH 4, and then extracted with CH2C12. The organic phase is washed with brine, dried (MgSO4), and then evaporated. Acid 1 is obtained as a tan solid (3.80 0.011 mol; 92% yield).

WO 98/23621 PCTIUS97/21785 62 1 H-NMR (300 MHz, CDC13): 5 7.82 1H, J 16 Hz, ArCH=C), 7.56 1H, ArH), 7.33 1H, ArH), 6.29 1H, J 16 Hz, C=CHCO2t-Bu), 3.72 2H, SCH2), 1.51 9H, C(CH3)3).

D. Synthesis of cephem IV" Cephem amine V' (15.04 0.035 mmol) is suspended under a nitrogen atmosphere in 65 mL THF. A solution of DCC in methylene chloride (1M, 36.2 mL, 0.036 mmol) is added, and the mixture allowed to stir for 5 minutes. Acid 1 (13.15 0.035 mmol) is added and the mixture is stirred for 1.5 hours. Ether (-30 mL) is added, and the solids (mostly DCU) are filtered off. The red-colored filtrate is evaporated to -25-30 mL and ether and pentane are added to precipitate the cephem product. The solid cephem is collected, washed with ether, and dried under vacuum to afford diester IV" (14.2 0.019 mmol; 54% yield).

1 H-NMR (300 MHz, CDC13): 8 7.53 1H, J 16 Hz, ArCH=C), 7.45-7.20 12H, ArH), 6.98 1H, Ph2CH), 6.35 1H, J 16 Hz, C=CHCO2t-Bu), 5.82 (dd, 1H, J 5, 8 Hz, R1R2CHNR3), 4.97 1H, J 5 Hz, CH(NR)(SR)), 4.37 2H, CH2C1), 3.76 1H, J 16 Hz, ArSCH2), 3.55 1H, J 16 Hz, ArSCH2), 3.40 1H, J 16 Hz, RSCH2R), 1.54 9H, C(CH 3 3 WO 98/23621 PCTIUS97/21785 63 E. Synthesis of diacid IV'" Diester IV" (0.760 1.00 mmol) is dissolved in 4 mL CH2C12 and 0.8 mL anisole. Trifluoroacetic acid (2 mL) is added, and the mixture is stirred for 4 hours. The solvents are evaporated, and the residue triturated with CH2Cl2/ether. The solid is collected, washed with EtOAc and then dried under vacuum. Diacid IV"' is obtained (0.420 0.780 mmol; 78% yield) as a light yellow solid.

1 H-NMR (300 MHz, DMSO): 8 9.30 1H, J 8 Hz, RC(O)NH), 8.07 1H, ArH), 7.72 1H, J 16 Hz, ArCH=C), 7.54 1H, ArH), 6.68 1H, J 16 Hz, C=CHCO2H), 5.71 (dd, 1H, J 5,8 Hz, R1R2CHNR3), 5.13 1H, J Hz, CH(NR)(SR)), 4.55 2H, CH2C1), 3.97 2H, ArSCH2), 3.70 1H, J 16 Hz, RSCH2R), 3.53 1H, J 16 Hz, RSCH2R).

F. Synthesis of cephem IA' Diacid IV"' (0.780 1.45 mmol) is dissolved in 3 mL methanol and 8 mL CH2C12. Thiopyridone III' (0.395 1.45 mmol) is added, and the mixture is stirred at room temperature for 4 hours. The flask is then placed in a refrigerator at 4°C overnight. The solvents are concentrated, and the product precipitated with ether. The solid is filtered, then suspended in EtOAc and stirred for 30 minutes. The product is then filtered, and the solid dried under vacuum. Cephem IA' is obtained as a tan solid (0.690 0.850 mmol; 59% yield of a mixture of two diastereomers).

WO 98/23621 PCTIUS97/21785 64 1 H-NMR (300 MHz, DMSO, partial): 8 9.31 1H, J 8 Hz, RC(0)NH), 8.67 2H, J 7 Hz, SpyrH), 7.98 2H, J 7 Hz, SpyrH), 8.07 1H, ArH), 7.72 1H, J 16 Hz, ArCH=C), 7.54 1H, ArH), 6.69 1H, J 16 Hz, C=CHCO2H), 5.70 (dd, 1H, J 5, 8 Hz, R1R2CHNR3), 5.14 1H, J 5 Hz, CH(NR)(SR)), 4.59-4.45 (in, 2H, CH2Spyr), 4.40-4.32 (in, 2H, pyrCH2R), 4.05- 3.95 (mn, 2H, ArCH2S), 1.38 9H, C(CH3)3).

G. Synthesis of cephem IA" Cephein IA' (0.605 0.747 mmol) is suspended in 3 mL CH2CI2, and 1 mL of trifluoroacetic acid is added. The solution is stirred for two hours and the solvents evaporated. The crude material is dissolved in CH2CI2 and precipitated with ether. The solids are collected, suspended in EtOAc and stirred for 30 minutes. The solids are collected and dried under vacuum (P205). Cephemn IA" is obtained (0.410 0.609 minol; 82% yield).

1 H-NMR (300 MHz, DMSO, partial): d 9.40-9.35 (in, 1H, RC(0)NH), 8.77- 8.72 (in, 2H, SpyrH), 8.23-8.18 (in, 2H, SpyrH), 8.12 1H, ArH), 7.77 1H, J 16 Hz, ArCH=C), 7.62 1H, ArH), 6.75 1H, J 16 Hz, C=CHCO2H), 5.68-5.61 (in, 1H, R1R2CHNR3), 5.11 1H, J 5 Hz, CH(NR)(SR)), 4.07- 3.97 (in, 2H, ArSCH2).

WO 98/23621 PCT/US97/21785 Following the general procedures and using the appropriate starting materials described above, the following additional compounds were prepared: x Y H R3 A-(L O S R 2 COOH R 4 WO 98/23621 WO 9823621PCTIUS97/21785 Cl~~ N.

-NH

2 cl Cl l

S.

Cl~~ NN-

CH

3 Cl 8 sCl- 3

N.N

Cl-I 0 2 C N 0

H

9s NN- NH2

S]

C0 2 I WO 98/23621 WO 9823621PCTIUS97/21785 67 WO 98/23621 WO 98/362 1PCTIUS97/21785 16

S",

CC

C0 2 17 Cl

OH

Cl *0 2 C N 0 18 Cl

OH

Cl 0 C0 2 19 S.,CH 3 Cl E) CH 3 Ici CH 3

CH

202 cl

NNH

2 Cl

CH

3 02 N 0 WO 98/23621 PCTfUS97/21785 69 21

CH-

3 Cl

CH

3 3 0 2 C N 0

HH

0 CH3 ol (D

H

2 TABLE: NMR DATA 7' H ArS N S 2 0 3'

CO

2

H

n) 4.40- 3.97 (br 9.40-9.35 8.12 8.77-8.72 4.80-4.60 bis TFA M+ 685 4.39 s) 7.77 J= 16) 8.23-8.18 4.40-4.24 salt 7.62 4.23-4.10 (m) 6.75 J 16) 3.10-3.00 (m) 5 5 2.80-2.65 (m) 4.41- 3.95- 9.24 7.47 8.62 J 6) 4.62-4.57 bis TFA MH+ 687 4.37 3.83 8) 7.45 8.05 J 6) 4.30-4.23 salt 2.84 J 7) 4.09-3.98 (m) 2.48 J 7) 3.10-2.98 (m) I' 1 12.80-2.64 4.41 4.09 (br D9.40-9 exch. 7.74 8.54 J 7) 4.50-4.40 bis TFA M 752 4.39 7.41 7.97 73-8.18 3.92-3.77 zwitterion 4.29 7.39 J 16) 3.49-3.21 (m) _6.39 S= 13) _6.39 j 16) 2.40-2.30 (m) co 3.4 (d j 4.90 J 5) 5.41 (dd, J 5, 4.37 3.96 9.23 J= 7.81 8.02 mono Na M* 654 4 17) 8) j =15) 8) 7.51 salt 3.26 j 3.88 7.33 j 16) zwitterion 17) j 15) 7.02 (s) j= 16) 3.45 I 4.94T j 5.41 (dd, j 5, 4.64 3.92- 9.23 J 7.82 7) mono Na M=626 17) 8) J =14) 3.83 8) 7.50 8.23 j= 7) salt 3.29 J 4.27 7.35 j 16) zwitterion (d =(14 6.53 16) 3.6 d j= .5 dj= .5 dd =5 4.65 3.95 (s)9.1 8.03 8.92 J 7) 3.42-3.32 (in) mono Na M 680 6 17) 8) j =14) 8) 7.68 j= 16) 8.33 j= 7) 2.20-1.90 salt 3.36 j= 4.44 7.56 zwitterion 17) j =14) 6.68 j= 16) 3.76 J 5.16 j 5.7 4.18 4.00 9.27 J= 8.10 8.69 j 6) 3.45 chloride M' 628 7 18) 5,8) 8) 7.57 8.00 j= 6) salt 3.56 j= 7.74 j 16) 18) 6.72 16) 3.45 J 4.85 j 5.40 J= 5) 4.28- 4.05 (br -oF -ue 7.83 7.75 (br s) -4.45S-437(in) zwitterion MW 754 8 16) 4.21 (mn) s) 7.55 J 16) 2.68 3.80-3.75 (in) sodium salt 3.24 J= 7.48(s) 16) 6.81 j 16) 3.83 J= 8) 3.6-3.2 (in) 4.92 j 5) 5.39 (dd, J= 5, 4.65 3.90- (d 8.58 j 7) mono Na -M 646 8) J 13) 3.70 (in) 8) 7.36 8.23 j 7) salt 9 4.25 zwitterion J =13) 3.75 J= 5.13 J 5.68 (dd, j 4.40 3.96 9.07 I 7.65 8.63 J 7) 7.65 (br s) chloride MH-1= 689 1 d8) 5,8) j =14) 8) 7.39 8.01 J 7) 5.20 salt 3.53(d, 4.3 (d .87(s) 18) J =14) 3.6-3.2 (in) 4.96 J 5.42 j 5) 4.42- 3.95 D 2 0 exch. 7.60 j 16) 8.62 J 7) 8.88 mono Na 4.19 (in) J=14) 8.00-7.07 (mn) 8.36 j 7) salt/ 113.78 6.38 J 16) zwitterion j 14) 3.76 J 5.15 j 5) 5.69 (dd, J 5, 4.46 3.98 9.34 J 8.07 9.34 J 6) 8.04 J= 8) sodium salt MW* 748 12 18) 8) J =10) 8) 7.71 j= 16) 8.14 j= 6) 7.46(d, j= 2) 3.56 j 4.44 7.55(Cs) 7.31 (dd, J 2, 18) j =10) 6.70 j= 16) 8) 3.74 J 5.15 (d,TJ= 5) 5.70 (dd, J 5, 4.36- 4.05- -934(T j- 7.86 7.83 7.80 j zwitterion 13 12) 8) 4.31 3.95 8) 7.56 2.75 7.68 J 2) chloride other 4.40-4.31 salt obscured 3.77 (s) (s) 3.75 j 5.14 j 5) 5.68 (dd,TI= 5, 4.40 4.02 9.35 j 7.87 8.89 j 7) 4.5 brs) -MWf 0 14 18) 8) J 14) 8) 7.71 8.01 J 7) 3.80 (br s) salt 3.47 J= 4.34 4.61 (s) 18) j =14) 3.50-3.20 5.13 J 5.66 (dd, j 4.40- 4.08 9.35 J 7.88 7.81 7.76 J 2) chlorid MW 829 8) 4.25 8) 7.68 2.73 7.66 j= 2) salt 4.9Cs) 4.545Cm) 3.80-3.60 (m) 2.70-2.50 (in) 2.48 (s) 3.69 j 5.12 J 5.64 (dd, J 6, 4.41 3.96 (br 9.28 J= 8.83 J= 8) 8.67 J 8) 4.51-4.45 (in) chloride MW 688 16 16) 8) j 13) s) 8) 7.50 8.08 J 8) 3.83-3.77 (mn) salt 3.48 j 4.35 7.47 (s) 16) 13) 3.88 j 8) 3.72 j 5.12 .66 (d 6, -4.41 -3.96 br- 9.30 Cd,- 7 8) 8.42 j 6) 86(dJ=8)4.48-4.42 I-Mo1rie Mi-P 714- 17 17) 8) J 11) s) 7.81 8.02 J 8) 3.81-3.75 (in) salt 3.49 j 4.34 7.56 J= 16) 17) j 11) 7.55(Cs) 6.81 J= 16) .86(d, 3.76 J 5.15 5) 5.66 (dd, 5, 4.42 4.03- 9.31 j 7.86 8.66 j 7) 4.53-4.45 (in) chloride M* 696 18 17) 8) j 12) 3.93 (in) 8) 7.61 8.02 j 7) 3.83-3.76 salt 3.51 j= 4.35 7.35 j 3) 17) 12) 7.24 j= 3) 3.74 (d j 1(d,j 5W 5.0(j 5, 4.39- 3.98 9.31 J= 7.82 7.80 7.79 j 1) chloride 3 19 16) 8) 4.29 (in) 8) 7.73 J 3) 2.74(Cs) 7.66 J 1) salt 3.49 J 7.63 4.46-4.35 (in) 16) 7.51 J 3) 4.38-4.31 (m) 3.76 (s) 2.64 (s) 230-218 (2 3.45 J= 4.85 J= 5) 5.41 (dd, J= 5, 4.42 obscured 9.22 J= 7.81 7.81 mono Na M' 712 17) 8) j =14) 8) 7.53 (d,j =16) salt 2 3.6(,j4.22 7.49 zwitterion 17) j =14) 6.83 j= 16) 3.95-3.87 48(dJ=5.4 (d 4.48 3.92 (9r -9.1-9(d,TJ= 8.10 7.83 4.37-4.25 (in) I M=83 5,8) J 13) s) 8) 7.52 3.95-3.85 zwitterion 4.34 7.46 J 16) 3.65-3.60 (in) J13) 7.03 J 16) 3.45-3.15 (in) 21 2.75 (s) 2.25-2.15 (in) 3.48 J 5.12 J= 5) 5.68 (dd, J= 5, 4.40- .4.06- 9.36 J 8.35 J= 4) 7.80 7.76 J 2) chloride MH 4 22 18) 8) 4.25 3.92 (in) 8) 7.87 7.61 J 2) salt 3.20 J 7.60 4.65-4.50 (m) 18) 3.70 J 4) 3.80-3.66 (in) 2.72 (s) 2.65 (s) (in)

Claims (10)

1. A compound of the formula X Y H- (X)M A-(L CH 2 -S-Q COOM wherein Q is an optionally substituted pyridinium group connected to the sulfur atom via a ring carbon atom; X is halogen; Y is hydrogen or halogen; A is COH, PO 3 H2, SOH or tetrazole; L' is a furylene group, a thienylene group, a C 2 -C 1 0 alkylene group, or a C2-C10 alkylene group interrupted by one or more groups independently selected from vinyl, S, SO, SO 2 SO2NH, H N/ or N H 0 n is 0 or 1; M is hydrogen, a negative charge or a carboxyl-protecting group; m is 0 or an integer selected so as to make the molecule as a whole electrically neutral; and x is a counter anion; or a pharmaceutically acceptable salt and/or a prodrug thereof. S:

2. A compound of the formula GG289A X Y e H R (X)m N-R 2CH 2 2 COOM R IA wherein. is halogen; Y.is-hydrogen:or-halegen :A.is-CQH, -OH, SO 3 H ar-tetrazolyl L' is a furylene group, a thienylene group, a C 2 -Co alkylene group, or a C 2 -C 10 alkylene group interrupted by one or more groups independently selected from vinyl, S, SO, SO 2 SO 2 NH, H O N or N H0 0 n is 0 or 1; R' and R 4 are each independently selected from hydrogen or C,-C 6 alkyl; R 2 is hydrogen, NH,, pyrrolidinyl, C 3 -C, 6 cycloalkyl, alkyl, C 2 -C 6 alkyl substituted by 60 6 one or more substituents independently selected from OH, NR'R 6 in which R 5 and R 6 are each independently hydrogen or alkyl, CO 2 H, morpholinyl, morpholinyl quaternized by a C,-C, 6 alkyl group, oxo, halogen, SOH, P0 3 H2, imidazolyl, imidazolyl substituted by 1-2 alkyl groups, tetrazolyl, tetrazolyl substituted by 1-2 alkyl 15 groups or N=CR' in which R 7 is a furan or thiophene ring optionally substituted by either -CO 2 H or -SO 3 H, phenyl or phenyl substituted by 1-3 substituents independently selected from OH, NR'R 6 in which R' and R 6 are as defined above, CO2H, morpholinyl, morpholinyl quaternized by a C,-C, 6 alkyl group, oxo, halogen, SO3H, P0 3 H 2 imidazolyl, imidazolyl substituted by 1-2 C,-C, 6 alkyl groups, tetrazolyl, tetrazolyl substituted by 1-2 C,-C, 6 alkyl groups or N=CR 7 in which R' is as defined above; M is hydrogen, a carboxyl- protecting group or a negative charge; x is a counter anion; and is 0 or an integer ~s~eprotecting group or a negative charge; x is a counter anion; and m is 0 or an integer GG289A 7 selected so as to make the molecule as a whole electrically neutral; or a pharmaceutically acc eptable salt and/or a prodrug thereof.

3. A compound of the formula e H R 3 XM. n 0 CH 2 NR COOMR4 IA wherein X is halogen; Y is hydrogen or halogen; A is C2'I P0 3 H 2 1 S 0 3 H or tetrazolyl; L' is a furylene group, a thienylene group, a C 2 -C 10 alkylene group or a C 2 -C 10 alkylene group interrupted by one or two groups independently selected from vinyl, S, So, SO 2 SO 2 NH, N* or ",NY *H 0 n isO0 or 1; R' and R' are each independently selected from hydrogen or C 1 -C 6 alkyl; R 2 is hydrogen, NH 2 pyrrolidinyl, CF-C 6 alkyl, C 1 -C 6 alkyl -substituted by one or two 15 substituents independently selected from OH, NR 5 R 6 in which R' and R 6 are each independently hydrogen or C 1 -C 6 alkyl, CO 2 H, morpholinyl, morpholinyl quaternized by a C 1 -C 6 alkyl group, oxo, halogen, SO 3 H, P0 3 H 2 tetrazolyl, CH 3 -l or N- H3 GG289A 77 in which R 7 is a furan or thiophene radical optionally substituted by a -CO 2 H or -SO 3 H group, phenyl or phenyl substituted by 1-2 substituents independently selected from OH, NR 5 R 6 in which R' and R 6 are as defined above, C2H, S3H P0 3 H 2 tetrazolyl, CH 3 halogen; M is hydrogen, a carboxyl-protecting group or a negative charge; X is a counter anion; and m is 0 or an integer selected so as to make the molecule as a whole electrically neutral; or a pharmaceutically acceptable salt and/or a prodrug thereof. x Y

4. A compound of claim 3 wherein S- is S Sn GG289A C] C] C0 2 H C] C0 2 H C1 C1 O=Sh C0 2 H C] C1 C0 2 H .C1 Cl C0 2 H S- C) C] o=s N C0 2 H C0 2 H S *0 C. C S 0 9* 0 C 99 4* C S C C C C 0 C 9 C. S C 9 .5

9. 9 C e.g. 99 0* *0 0 0 Ss C] C1 C1 N, C] Cl 0 HO 2 C' N 0 C0 2 H H C1 C] C1 C) or H0 2 C H 0 C2 5. A compound of claim 3 or claim 4 wherein R 3 C1 S Co Cl 2 H H0 2 C' O=S=O C02H GG289A H 3 C N-NH2 CH 3 N NH 2 CH 3 CH 3 OH CH 3 0 N -N 0 00 0 00 0 g eS g 0 C. CS S S S 00 0 0 C C O C O 0 e 0@ g 0 (D 0Y""ii iO S03H CH OOH NH 3 OH CH 3 CH 3 HCC CH 3 CH 3 OH \7 C0 2 H CH 3 CH 3 CH 3 N- CH- CH 3 r-N; 0 OHY NH 3 OH CH 3 N CH 3 OH \N C0 2 H (D OH 6. A compound of the formula GG289A x Y I H S,,y 0 CH 2 -S-Q COOM 0D x y wherein S- A- (L) 1 x y C1 l and Q are as defined below: Q H 3 C CH 3 H 2 CH 3 S- C] Cl C0 2 H G C2 89A C' C0 2 H C N- NH 2 C (D N In C0 2 H a a. a. C] C0 2 H -o- CH 3 -o OH C] CH- 3 H0 2 C' GG289A Cl C0 2 H C-( ,N-'TH2 Cl Cl C0 2 H C0 2 H 0 H 9 NH2 OH C] C0 2 H OH -0 C0 2 H C0 2 H -0 0 SO.-H I N G G 2 S A 33 Cl I C) C0 2 H Cl-b -D ,*r OH- CH-1 (in) C] ci C0 2 H- 0D CH-- CH, c H a C] OH N o0//S 'N 1 C0 2 H a a a CR 3 -0 0D C H 4N CH3 C H1 C0 2 H -0H C0 2 H GG29A OOH H0 2 C~ N 0 H (r aC C] s C0 2 H CH 3 D CH- Cl1 3 N- NCH3 a a a. a a Cl H0 2 C~ N 0 H Ho 2 C N 0 )CH CH, N-NH 2 CH 3 CH 3 -00 CH 3 GG289A C] 0 C o NH C0 2 H C]' 3 C (D DCH, \NNCH CM 3 N CN 9 9 *9*9 9*9* 99 9 9 0~ C02 and M is hydrogen, a negative charge, or a carboxyl-protecting group; x is a counter anion; and mn is 0 or an integer selected so as to na'ke the. molecule as a whole electrically neutral; or a pharmaceutically acceptable salt and/or a prodrug thereof. 7. The compound of the formula 0 COON wherein X is a counter anion; or a pharmaceutically acceptable salt and/or a prodr-ug thereof. GG289A 8. The compound of the formula C0 HO 0 C) C0 2 H N-NH 2 wherein X is a counter anion; or a phaiaceuti]cally acceptable salt and/or a prodi-ug thereof.- S S 5. S 5* S S *SS S S S S S *S.S 55 S S S *S S S *5*5*5 S S 55 S S S S 9. The compound of the formula OH Cl S G ,wherein X is a counter anion: or a pharmaceutically acceptable salt andlor a prodrug thereof. The compound of the formnula GG289A 87 wherein X is a counter anion; or a pharmnaceutically acceptable salt andlor a prodrug, thereof.

11. The compound of the formula C) 0 S K H H S NO 5, CH-I 0 CI 0 COOH J NH 2 CH.- 0 wherein X is a counter anion; or a pharmaceutically acceptable salt and/or a prodr-ug thereof.

12. The compound of the formula S *S S CH 2 CH 3 CH 3 N "yOH 0 0 1 5 wherein X is a counter anion; or a pharmaceutically acceptable salt and/or a prodrug thereof. 88

13. A pharmaceutical composition comprising an effective antibacterial amount of a compound of any one of claims 1 to 12 and a pharmaceutically acceptable carrier or excipient.

14. A method of treating a bacterial infection with comprises administering to a host afflicted with such infection an effective antibacterial amount of a compound of any one of claims 1 to 12. A method of treating a bacterial infection caused by a strain of methicillin- resistant Staphylococcus aureus which comprises administering to a host afflicted with such infection an effective antibacterial amount of a compound of any one of claims 1 to 12.

16. A compound substantially as hereinbefore described with reference to 15 any one of the examples DATED: 17 April 2001 20 PHILLIPS ORMONDE FITZPATRICK Attorneys For: BRISTOL-MYERS SQUIBB COMPANY 0 0