AU2005202802A1 - Cephem compounds - Google Patents

Description

Regulation 3.2

AUSTRALIA

Patents Act 1990 COMPLETE SPECIFICATION FOR A STANDARD PATENT

(ORIGINAL)

Name of Applicants: Astellas Pharma, Inc., of 3-11, Nihonbashi-Honcho 2-chome, Chuo-ku, Tokyo 103-8411, JAPAN AND Wakunaga Pharmaceutical Co., Ltd., of 5-36, Miyahara 4-chome, Yodogawa-ku, Osaka-shi, Osaka 532-0003, JAPAN Actual Inventors: YAMANAKA, Toshio OKUDA, Shinya TODA, Ayako MURANO, Kenji OHKI, Hidenori KAWABATA, Kohji INOUE, Satoshi MISUMI, Keiji ITOH, Kenji SATOH, Kenji Address for Service: Invention Title: DAVIES COLLISON CAVE, Patent Attorneys, of 1 Nicholson Street, Melbourne, Victoria 3000, Australia "Cephem compounds" Details of Associated Provisional Application Nos: 2004903529 2004903705 The following statement is a full description of this invention, including the best method of performing it known to us: Q:\OPER\Kbm\2005\Jun\12628860 174.doc 27/6/05 P:\OPER\Kbm\specis\12628860 specidoc

DESCRIPTION

o CEPHEM COMPOUNDS TECHNICAL FIELD ;The present invention relates to new cephem compounds r- 5 and pharmaceutically acceptable salts thereof. More particularly, the present invention relates to new cephem compounds and pharmaceutically acceptable salts thereof, Swhich have antimicrobial activities, to processes for 00 preparation thereof, to pharmaceutical composition comprising the same, and to a method for treating infectious Sdiseases in human being and animals.

BACKGROUND ART Some cephem compounds are disclosed in W000/32606 and W003/078440.

DISCLOSURE OF INVENTION Advantageously at least one embodiment of the present invention may provide novel cephem compounds and pharmaceutically acceptable salts thereof, which are highly active against a number of pathogenic microorganisms.

Another advantage of at least one embodiment of the present invention is that processes for the preparation of said cephem compounds and salts thereof may be provided.

A further advantage of at least one embodiment of the present invention is that a pharmaceutical composition comprising, as an active ingredient, said cephem compounds or their pharmaceutically acceptable salts may be provided.

Still further, an advantage of at least one embodiment of the present invention is that a method for treating infectious diseases caused by pathogenic microorganisms, which comprises administering said cephem compounds to infected human being or animals may be provided.

The cephem compounds of the present invention are novel and can be represented by the following general P:\OPER\Kbm\specis\12628860 speci.doc formula H3 CCH3 C O R 1

N

11

S

II

N 7- C- CONH 2X ,N -N CH 2

-R

3

[I]

R S o

COO

0 wherein

R

1 is carboxy or protected carboxy,

R

2 is amino or protected amino, and 3 is

R

6

R

6 R 4 R R 7

N

r N R V R 7 R R 8 or

R

8

R

7 wherein

R

4 is hydrogen, amino or protected amino,

R

5 is -(X)n-A-R 9 or a saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms, wherein X is -NHCO- or -NR 1 0 (wherein R 10 is hydrogen or lower alkyl), A is lower alkylene,

R

9 is amino or protected amino, and n is 0 or 1,

R

6 and R 8 are each hydrogen, amino, protected amino or -(X)n-A-R 9 wherein X, A, R 9 and n are each as defined above, and

R

7 is hydrogen, amino(lower)alkyl or protected amino(lower)alkyl.

As to the compound the following points are to be noted.

P:\OPER\Kbm\specis\12628860 specidoc That is, the compound includes syn isomer (Z form), anti isomer (E form) and a mixture thereof. Syn C isomer (Z form) means one geometrical isomer having the Spartial structure represented by the following formula:

H

3 C

CH

3 c N-O R1

II

O R2S/ N c 5 R S Swherein R 1 and R 2 are each as defined above, Sand anti isomer (E form) means the other geometrical isomer Shaving the partial structure represented by the following formula:

H

3 C CH 3 R O-N

II

N- C- CO- R 2 SN wherein R 1 and R 2 are each as defined above, and all of such geometrical isomers and mixtures thereof are included within the scope of this invention.

In the present specification and claims, the partial structure of these geometrical isomers and mixtures thereof are represented for convenience' sake by the following formula:

H

3 C CH 3 O R 1

N

II

R S

N

wherein R 1 and R 2 are each as defined above.

The cephem compound of the present invention can be prepared by the following processes as illustrated in the following.

P:\OPER\Kbm\specis\1262BB6O specidoc Process 1

CH

2 R 3 coo

E

H

3

CXCH~{

N

11 N-\C COOH R 2X

[III]

or its reactive derivative at the carboxy group, or a salt thereof

[III]

or its reactive derivative at the amino group, or a salt thereof

H

3 C CH 3 0XRl

N

11 S

N-\V-C-CONHY

N- CH 2

R

R s o

[I]

or a salt thereof P:\OPER\Kbm\speCiS\12628860 specidoc Process 2

H-

3 C CH 3 0x R

N

11

N\CCNH

7 R 2X'

IN

CH

2 R 3a Ia] or a salt thereof Elimination reaction of the I amino protecting group

H

3 C CH 3 O R1

N

N-CCONH'

R 2XI

N

,5 CH 2

-R

3 b

COO®-

[Ib] or a salt thereof P:\OPER\Kbm\speciu\12628860 specidoc Process 3

R

3 c

[V]

or a salt thereof

[IV]

or a salt thereof

CH

2

R

3

[I]

or a salt thereof wherein R 1

R

2 and R 3 are each as defined above,

R

3 a is R~a

-N

I-)

R8a Ra K- R8a

N

Ra R8a wherein R4a is hydrogen or protected amino,

R

5 a is -(X)n-A-R 9 a wherein X, A and n are each as defined above, and R9a is protected amino, R6a and R 8 a are each hydrogen, protected amino or -(X),-A-R9a wherein X, A, R 9 a and n are each as defined above, and P:\OPER\Kbm\apecis\12628860 specidoc

R

7 a is hydrogen or protected amino(lower)alkyl, O with the proviso that R a, R 7 a and R8a are not hydrogen at C the same time,

SR

3 b is R6b R6b R 4b R b

R

7 b N- N N R5 R R o r

N

0 Rb I "R7b R8b or R8b R7b l wherein

R

4 b is hydrogen or amino,

R

5 b is -(X)n-A-NH 2 wherein X, A and n are each as defined above,

R

6 b and R 8 b are each hydrogen, amino or -(X)n-A-NH 2 wherein X, A and n are each as defined above, and

R

7 b is hydrogen or amino(lower)alkyl, with the proviso that R 6 b, R 7 b and R 8 b are not hydrogen at the same time,

R

3 c is a compound of the formula selected from

R

6

R

6 R4 6

R

R RR N N N N N i -R 8

R

5

>NR

7

R

8 and N

R

8

R

7 wherein R 4

R

5

R

6

R

7 and R 8 are each as defined above,

R

11 is carboxy or protected carboxy, and Y is a leaving group.

The starting compounds [II] and [IV] can be prepared by the following processes.

P:\OPER\Kbm\specis\1262886D specidoc Process A R 1 T(

CH

2

-Y

0 R 13

[VI]I

or a salt thereof

R

3 c or a salt thereof

CH

2

-R

3 .*zE

[VII]

or a salt thereof .zCH 2 -R 3

[III

or a salt thereof P:\OPER\Kbm\specis\12628860 speci~doc Process B

H

3 C CH 3 0x 0 R 1

N

s II

H

2 N- N C--COOH -N -CH2Y R2 N C RR S 00 C [VIII] [IX] or its reactive or its reactive D derivative at the derivative at the Samino group, carboxy group, or a salt thereof or a salt thereof

H

3 C CH 3 OXR1

N

II

N-TR2C-CONH- CH2 2X SN 4 N -CH 2

-Y

R S 0 11

[IV]

or a salt thereof wherein R 1

R

2

R

3

R

11 and Y are each as defined above, ZO is an anion,

R

12 is protected amino, and

R

13 is protected carboxy.

The starting compound can be prepared by the methods disclosed in the Preparations 1A-18A described later or similar manners thereto.

In the above and subsequent descriptions of this specification, suitable examples of the various definitions are explained in detail as follows.

P:\OPER\Kbm\specis\12628860 specidoc The term "lower" is used to mean a group having 1 to S6, preferably 1 to 4, carbon atom(s), unless otherwise indicated.

Suitable "lower alkyl" and "lower alkyl" moiety in "amino(lower)alkyl" and "protected amino(lower)alkyl" include straight or branched alkyl having 1 to 6 carbon atom(s), such as methyl, ethyl, propyl, isopropyl, butyl, Sisobutyl, sec-butyl, tert-butyl, pentyl, tert-pentyl and

C

hexyl, in which more preferred one is CI-C 4 alkyl.

r 10 Suitable "amino(lower)alkyl" includes amino(C 1

C

s )alkyl such as aminomethyl, 1-aminoethyl, 2-aminoethyl, 1c aminopropyl, 2-aminopropyl, 3-aminopropyl, 4-aminobutyl, aminopentyl and 6-aminohexyl, in which more preferred one is amino(C 1

-C

4 alkyl.

Suitable "lower alkylene" includes straight or branched alkylene having 1 to 6 carbon atoms, such as methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene and propylene, in which more preferred one is straight alkylene having 1 to 3 carbon atoms.

Suitable "saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms" includes azetidinyl 1-azetidinyl and 3-azetidinyl), pyrrolidinyl 1-pyrrolidinyl and 3-pyrrolidinyl), imidazolidinyl 1imidazolidinyl and 4-imidazolidinyl), piperidinyl 1piperidinyl and 4-piperidinyl) and piperazinyl 1piperazinyl), in which more preferred one is saturated 4- to 6-membered heterocyclic group containing 1 to 4 nitrogen atoms.

Suitable "amino protecting group" in "protected amino" and "protected amino(lower)alkyl" includes an acyl group as mentioned below, substituted or unsubstituted aryl(lower)alkylidene benzylidene, P:\OPER\Kbm\speciS\12628860 specidoc hydroxybenzylidene, etc.], aryl(lower)alkyl such as mono-, di- or triphenyl(lower)alkyl benzyl, phenethyl, benzhydryl, trityl, etc.], and the like.

;ZSuitable "acyl" includes lower alkanoyl formyl, acetyl, propionyl, hexanoyl, pivaloyl, etc.], mono(or di or tri)halo(lower)alkanoyl chloroacetyl, trifluoroacetyl, etc.], lower alkoxycarbonyl methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, tert- 00 pentyloxycarbonyl, hexyloxycarbonyl, etc.], carbamoyl, aroyl benzoyl, toluoyl, naphthoyl, etc.], aryl(lower)alkanoyl phenylacetyl, phenylpropionyl, etc.], aryloxycarbonyl phenoxycarbonyl, naphthyloxycarbonyl, etc.], aryloxy(lower)alkanoyl phenoxyacetyl, phenoxypropionyl, etc.], arylglyoxyloyl phenylglyoxyloyl, naphthylglyoxyloyl, etc.], aryl(lower)alkoxycarbonyl which optionally substituted by suitable substituent(s) benzyloxycarbonyl, phenethyloxycarbonyl, p-nitrobenzyloxycarbonyl, etc.], and the like.

Preferable examples of "amino protecting group" include aryl(lower)alkyl and acyl, in which more preferred ones are aryl(lower)alkyl, lower alkanoyl and lower alkoxycarbonyl, and particularly preferred ones are mono-, di- or triphenyl(C 1

-C

6 )alkyl, C 1 -C6 alkanoyl and (C 1

C

6 )alkoxycarbonyl.

Preferable examples of "protected amino" and "protected amino" moiety in "protected amino(lower)alkyl" include aryl(lower)alkylamino and acylamino, in which more preferred ones are aryl(lower)alkylamino, lower alkanoylamino and lower alkoxycarbonylamino, and particularly preferred ones are mono-, di- or triphenyl(C 1

C

6 )alkylamino, Cj-C 6 alkanoylamino and (C 1

C

6 )alkoxycarbonylamino.

P:\OPER\Kbm\specis\12628860 specidoc Suitable "protected carboxy" includes an esterified Scarboxy group and the like, and concrete examples of the C ester moiety in said esterified carboxy group include the Sones such as lower alkyl ester methyl ester, ethyl ester, propyl ester, isopropyl ester, butyl ester, isobutyl ester, tert-butyl ester, pentyl ester, hexyl ester, 1cyclopropylethyl ester, etc.] which may have suitable Ssubstituent(s), for example, lower alkanoyloxy(lower)alkyl 00 C ester acetoxymethyl ester, propionyloxymethyl ester, N 10 butyryloxymethyl ester, valeryloxymethyl ester, pivaloyloxymethyl ester, l-acetoxyethyl ester, 1- C propionyloxyethyl ester, 2-propionyloxyethyl ester, hexanoyloxymethyl ester, etc.], lower alkanesulfonyl(lower)alkyl ester 2-mesylethyl ester, etc.] or mono(or di or tri)halo(lower)alkyl ester 2iodoethyl ester, 2,2,2-trichloroethyl ester, etc.]; lower alkenyl ester vinyl ester, allyl ester, etc.]; lower alkynyl ester ethynyl ester, propynyl ester, etc.]; aryl(lower)alkyl ester which may have suitable substituent(s) benzyl ester, 4-methoxybenzyl ester, 4-nitrobezyl ester, phenethyl ester, trityl ester, benzhydryl ester, bis(methoxyphenyl)methyl ester, 3,4dimethoxybenzyl ester, 4-hydroxy-3,5-di-tert-butylbenzyl ester, etc.]; aryl ester which may have suitable substituent(s) phenyl ester, 4-chlorophenyl ester, tolyl ester, 4-tert-butylphenyl ester, xylyl ester, mesityl ester, cumenyl ester, etc.]; and the like.

Preferable examples of "protected carboxy" include lower alkoxycarbonyl and aryl(lower)alkoxycarbonyl which may have suitable substituent(s), in which more preferred one is

(C

1

-C

6 alkoxycarbonyl.

Suitable "leaving group" includes halogen chlorine, bromine, iodine, etc.] or acyloxy such as P:\OPER\Kbm\speci9\12628860 specidoc Sarylsulfonyloxy benzenesulfonyloxy, tosyloxy, etc.], Slower alkylsulfonyloxy mesyloxy, etc.], lower alkanoyloxy acetyloxy, propionyloxy, etc.], and the Slike.

Suitable "anion" includes formate, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate, toluenesulfonate, chloride, bromide, iodide, sulfate, hydrogensulfate, phosphate, and the like.

C Suitable pharmaceutically acceptable salts of the compound are conventional non-toxic salts and include, §for example, a salt with a base or an acid addition salt such as a salt with an inorganic base, for example, an alkali metal salt sodium salt, potassium salt, etc.], an alkaline earth metal salt calcium salt, magnesium salt, etc.], an ammonium salt; a salt with an organic base, for example, an organic amine salt trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, etc.]; an inorganic acid addition salt hydrochloride, hydrobromide, sulfate, hydrogensulfate, phosphate, etc.]; an organic carboxylic or sulfonic acid addition salt formate, acetate, trifluoroacetate, maleate, tartrate, citrate, fumarate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc.]; and a salt with a basic or acidic amino acid arginine, aspartic acid, glutamic acid, etc.].

The preferred embodiment of the cephem compound of the present invention represented by the general formula is as follows.

The compound of the formula wherein

R

1 is carboxy or esterified carboxy,

R

2 is amino, aryl(lower)alkylamino or acylamino, P:\OPER\KbM\SPeCiS\12628860 specidoc Ris hydrogen, amino, aryl(lower)alkylamino or acylamino, R9 is amino, aryl(lower)alkylamino or acylamino, R 6 and R 8 are each hydrogen, amino, aryl(lower)alkylamino, ;Z acylamino or W(X)-A-R 9 wherein X, A, R 9 and n are each as defined above, and R 7is hydrogen, amino(lower)alkyl, aryl (lower) alkylamino (lower) alkyl or acylamino (lower) alkyl, 00i or a pharmaceutically acceptable salt thereof.

The compound of above wherein R1 is carboxy or (Cl-C 6 )alkoxycarbonyl, R 2 is amino, mono-, di- or triphenyl(C 1 -CG)alkylamino, CI-C 6 alkanoylamino or (C C 6 a lkoxyc arbonyl amino, R 4 is hydrogen, amino, mono-, di- or triphenyl (Cl-

C

6 alkylamino, Cj-C 6 alkanoylamino or (Cl-

C

6 alkoxycarbonylamino, R 5 is -(X)n-A-R9 or a saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms, wherein X is -NHCO- or -NR 1 0 (wherein R 10 is hydrogen or Cj-C 6 alkyl) A is Cl-C 6 alkylene,

R

9 is amino, mono-, di- or triphenyl(Cl-C 6 )alkylamino, Cl-C 6 alkanoylamino or (Cl-C 6 )alkoxycarbonylamino, and n is 0 or 1, R 6 and R8 are each hydrogen, amino, mono-, di- or triphenyl- (Cl-C 6 )alkylamino, Cl-C 6 alkanoylamino, (Cl-C 6 )alkoxycarbonylamino or 9 wherein X, A, R 9 and n are each as defined above, and R 7 is hydrogen, amino (Cl-C 6 alkyl, mono-, di- or triphenyl (Cl-C 6 alkylamino (Cl-C 6 alkyl, Cj-C 6 alkanoylamino(Cl-C 6 )alkyl or (C 1 CO) alkoxycarbonylamino (Cl CO alkyl, or a pharmaceutically acceptable salt thereof.

P:\OPER\Kbm\specis\12628860 specidoc The compound of the formula wherein SR3 is

R

4 ;Z -N

R

wherein R 4 and R 5 are each as defined in the formula or a pharmaceutically acceptable salt thereof.

00 C The compound of above wherein 4 C-q R is hydrogen or amino, and Q R is -(X)n-A-R 9 or piperazinyl, C( wherein X is -NHCO- or -NR 10 (wherein R 10 is hydrogen or lower alkyl), A is lower alkylene,

R

9 is amino, and n is 0 or 1, or a pharmaceutically acceptable salt thereof.

The processes for preparing the compound of the present invention are explained in detail in the following.

Process 1 The compound or a salt thereof can be prepared by reacting the compound [II] or its reactive derivative at the amino group, or a salt thereof with the compound [III] or its reactive derivative at the carboxy group, or a salt thereof.

Suitable reactive derivative at the amino group of the compound [II] includes Schiff's base type imino or its tautomeric enamine type isomer formed by the reaction of the compound [II] with a carbonyl compound such as aldehyde, ketone and the like; a silyl derivative formed by the reaction of the compound [II] with a silyl compound such as bis(trimethylsilyl)acetamide, mono(trimethylsilyl)acetamide N-(trimethylsilyl)acetamide], bis(trimethylsilyl)urea P:\OPER\Kbm\specis\12628860 specidoc and the like; a derivative formed by the reaction of the Ocompound [II] with phosphorus trichloride or phosgene.

Suitable salts of the compound [II] and its reactive Sderivative can be referred to the ones as exemplified for the compound Suitable reactive derivative at the carboxy group of the compound [III] includes an acid halide, an acid anhydride, an activated amide, and an activated ester. A C suitable example of the reactive derivatives may be an acid chloride; an acid azide; a mixed acid anhydride with an acid Ssuch as substituted phosphoric acid dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoric acid, halogenated phosphoric acid, etc.], dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, sulfuric acid, alkanesulfonic acid methanesulfonic acid, etc.], aliphatic carboxylic acid acetic acid, propionic acid, butyric acid, isobutyric acid, pivalic acid, pentanoic acid, isopentanoic acid, 2-ethylbutyric acid, trichloroacetic acid, etc.] and aromatic carboxylic acid benzoic acid, etc.]; a symmetrical acid anhydride; an activated amide with imidazole, 4-substituted imidazole, dimethylpyrazole, triazole or tetrazole; an activated ester cyanomethyl ester, methoxymethyl ester, dimethyliminomethyl [(CH 3 2 ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenyl thioester, p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester, pyranyl ester, pyridyl ester, piperidyl ester, 8quinolyl thioester, etc.]; or an ester with an N-hydroxy compound N,N-dimethylhydroxylamine, l-hydroxy-2-(1H)pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide, Nhydroxy-1H-benzotriazole, etc.]. These reactive derivatives P:\OPER\Kb\specis\12628860 specidoc can optionally be selected from them according to the kind Sof the compound [III] to be used.

Suitable salts of the compound [III] and its reactive Sderivative can be referred to the ones as exemplified for 5 the compound The reaction is usually carried out in a conventional solvent such as water, alcohol methanol, ethanol, Setc.], acetone, dioxane, acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine or any other organic solvent Swhich does not adversely affect the reaction. These conventional solvents may also be used in a mixture with water.

In this reaction, when the compound [III] is used in free acid form or its salt form, the reaction is preferably carried out in the presence of a conventional condensing agent such as N,N'-dicyclohexylcarbodiimide; N-cyclohexyl- N'-morpholinoethylcarbodiimide; N-cyclohexyl-N'-(4diethylaminocyclohexyl)carbodiimide; N,N'diethylcarbodiimide; N,N'-diisopropylcarbodiimide; N-ethyl- N'-(3-dimethylaminopropyl)carbodiimide; N,N'-carbonyl-bis- (2-methylimidazole); pentamethyleneketene-N-cyclohexylimine; diphenylketene-N-cyclohexylimine; ethoxyacetylene; 1-alkoxyl-chloroethylene; trialkyl phosphite; ethyl polyphosphate; isopropyl polyphosphate; phosphorus oxychloride (phosphoryl chloride); phosphorus trichloride; thionyl chloride; oxalyl chloride; lower alkyl haloformate ethyl chloroformate, isopropyl chloroformate, etc.], triphenylphosphine; 2-ethyl-7-hydroxybenzisoxazolium salt; 2-ethyl-5-(m-sulfophenyl)isoxazolium hydroxide intramolecular salt; 1-(p-chlorobenzenesulfonyloxy)-6chloro-lH-benzotriazole; so-called Vilsmeier reagent prepared by the reaction of N,N-dimethylformamide with P:\OPER\Kbm\specis\1262886O specidoc thionyl chloride, phosgene, trichloromethyl chloroformate, Sphosphorus oxychloride, etc.; and the like.

The reaction may also be carried out in the presence Sof an inorganic or organic base such as an alkali metal bicarbonate, tri(lower)alkylamine, pyridine, N- (lower)alkylmorpholine, N,N-di(lower)alkylbenzylamine, and the like.

SThe reaction temperature is not critical, and the 00 c reaction is usually carried out under cooling to warming.

S 10 Process 2 The compound [Ib] or a salt thereof can be prepared by subjecting the compound [Ia] or a salt thereof to elimination reaction of the amino protecting group.

Elimination reaction is carried out in accordance with a conventional method such as hydrolysis, reduction and the like.

The hydrolysis is preferably carried out in the presence of a base or an acid including Lewis acid.

Suitable base includes an inorganic base and an organic base such as an alkali metal sodium, potassium, etc.], an alkaline earth metal magnesium, calcium, etc.], the hydroxide or carbonate or hydrogencarbonate thereof, trialkylamine trimethylamine, triethylamine, etc.], picoline, diazabicyclo[4.3.0]non-5-ene, 1,4-diazabicyclo[2.2.2]octane, 1,8-diazabicyclo[5.4.0]undec-7-ene, and the like.

Suitable acid includes an organic acid formic acid, acetic acid, propionic acid, trichloroacetic acid, trifluoroacetic acid, etc.], and an inorganic acid hydrochloric acid, hydrobromic acid, sulfuric acid, hydrogen chloride, hydrogen bromide, etc.].

The elimination using Lewis acid such as trihaloacetic acid trichloroacetic acid, trifluoroacetic acid, P:\OPER\Kbm\specis\12628860 specidoc etc.], and the like is preferably carried out in the O presence of cation trapping agents anisole phenol etc.].

;The reaction is usually carried out in a solvent such as water, alcohol methanol, ethanol, etc.], methylene chloride, tetrahydrofuran, a mixture thereof or any other solvent which does not adversely influence the reaction. A liquid base or acid can be also used as a solvent.

00 C The reaction temperature is not critical and the reaction is usually carried out under cooling to warming.

SThe reaction method applicable for the elimination reaction may include chemical reduction and catalytic reduction.

Suitable reducing reagents to be used in chemical reduction are a combination of a metal tin, zinc, iron, etc.] or metallic compound chromium chloride, chromium acetate, etc.] and an organic acid or inorganic acid formic acid, acetic acid, propionic acid, trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid, hydrobromic acid, etc.].

Suitable catalysts to be used in catalytic reduction are conventional ones such as platinum catalysts platinum plate, spongy platinum, platinum black, colloidal platinum, platinum oxide, platinum wire, etc.], palladium catalysts spongy palladium, palladium black, palladium oxide, palladium on carbon, colloidal palladium, palladium on barium sulfate, palladium on barium carbonate, etc.], nickel catalysts reduced nickel, nickel oxide, Raney nickel, etc.], cobalt catalysts reduced cobalt, Raney cobalt, etc.], iron catalysts reduced iron, Raney iron, etc.], copper catalysts reduced copper, Raney copper, Ullman copper, etc.] and the like.

The reduction is usually carried out in a conventional P:\OPER\Kbm\specis\12628860 speci.doc solvent which does not adversely influence the reaction such O as water, methanol, ethanol, propanol, N,N-dimethylformamide or a mixture thereof.

SAdditionally, in case that the above-mentioned acids to be used in chemical reduction are liquid, they can also be used as a solvent.

Further, a suitable solvent to be used in catalytic Sreduction may be the above-mentioned solvent, and other 00 C conventional solvent such as diethyl ether, dioxane, S 10 tetrahydrofuran, etc., or a mixture thereof.

The reaction temperature of this reduction is not critical and the reaction is usually carried out under cooling to warming.

Process 3 The compound or a salt thereof can be prepared by reacting the compound [IV] or a salt thereof with the compound or a salt thereof, optionally followed by deprotection.

Suitable salt of the compounds [IV] can be referred to the ones as exemplified for the compound Suitable salts of the compounds include an organic acid addition salt formate, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc.], an inorganic acid addition salt hydrochloride, hydrobromide, sulfate, phosphate, etc.] and the like.

The present reaction may be carried out in a solvent such as water, phosphate buffer, acetone, chloroform, acetonitrile, nitrobenzene, methylene chloride, ethylene chloride, formamide, N,N-dimethylformamide, methanol, ethanol, diethyl ether, tetrahydrofuran, dimethyl sulfoxide, or any other organic solvent which does not adversely affect the reaction, preferably in ones having strong polarities.

P:\OPER\Kbm\9pecis\12628860 specidoc Among the solvents, hydrophilic solvents may be used in a O mixture with water. When the compound is liquid, it can C also be used as a solvent.

;The reaction is preferably conducted in the presence of a base, for example, an inorganic base such as alkali metal hydroxide, alkali metal carbonate, alkali metal hydrogencarbonate, an organic base such as trialkylamine, and the like.

00 The reaction temperature is not critical, and the C( 10 reaction is usually carried out at ambient temperature, Sunder warming or under heating. The present reaction is preferably carried out in the presence of alkali metal halide sodium iodide, potassium iodide, etc.], alkali metal thiocyanate sodium thiocyanate, potassium thiocyanate, etc.], and the like.

When R 11 is protected carboxy, the compound or a salt thereof can be prepared by subjecting the compound obtained above to elimination reaction of the carboxy protecting group.

Elimination reaction is carried out in a similar manner to the reaction in the aforementioned Process 2, and therefore the reagents to be used and reaction conditions solvent, reaction temperature, etc.) can be referred to those of Process 2.

Processes A and B for the preparation of the starting compounds are explained in detail in the following.

Process A-(i) The compound [VII] or a salt thereof can be prepared by reacting the compound [VI] or a salt thereof with the compound or a salt thereof.

This reaction can be carried out in a similar manner to the reaction in the aforementioned Process 3, and therefore the reagents to be used and reaction conditions P:\OPER\Kbm\SpeCiS\12628860 specidoc solvent, reaction temperature, etc.) can be referred to those of Process 3.

cq Anion ZO may be the one derived from a leaving group ;Y and may be the other one converted therefrom by a conventional method.

Process A-(ii) The compound [II] or a salt thereof can be prepared by 00 subjecting the compound [VII] or a salt thereof to c~ theaminoprotecting groups in R' 2 elimination reaction of the amino protecting groups in R 12 c13 V 10 and the carboxy protecting group in R 13 This reaction can be carried out in a similar manner to the reaction in the aforementioned Process 2, and therefore the reagents to be used and reaction conditions solvent, reaction temperature, etc.) can be referred to those of Process 2.

Process B The compound [IV] or a salt thereof can be prepared by reacting the compound [VIII] or its reactive derivative at the amino group, or a salt thereof with the compound [IX] or its reactive derivative at the carboxy group, or a salt thereof.

This reaction can be carried out in a similar manner to the reaction in the aforementioned Process 1, and therefore the reagents to be used and reaction conditions solvent, reaction temperature, etc.) can be referred to those of Process i.

The compounds obtained by the above processes can be isolated and purified by a conventional method such as pulverization, recrystallization, column chromatography, reprecipitation, and the like.

It is to be noted that the compound and other compounds may include one or more stereoisomer(s) such as optical isomer(s) and geometrical isomer(s) due to P:\OPER\Kbm\specis\12628860 specidoc asymmetric carbon atom(s) and double bond(s), and all of O such isomers and mixtures thereof are included within the scope of this invention.

SThe compounds and pharmaceutically acceptable salts thereof include solvates enclosure compounds hydrate, etc.)].

The compound and pharmaceutically acceptable salts Sthereof are novel and exhibit high antimicrobial activity, 00 C inhibiting the growth of a wide variety of pathogenic S 10 microorganisms including Gram-positive and Gram-negative Smicroorganisms and are useful as antimicrobial agents.

C Now in order to show the utility of the compound the test data on MIC (minimal inhibitory concentration) of a representative compound of this invention are shown in the following.

Test method: In vitro antibacterial activity was determined by the two-fold agar-plate dilution method as described below.

One loopful of an overnight culture of each test strain in Trypticase-soy broth (106 viable cells per ml) was streaked on heart infusion agar (HI-agar) containing graded concentrations of representative test compound, and the minimal inhibitory concentration (MIC) was expressed in jig/ml after incubation at 37 0 C for 20 hours.

Test compound Compound 3-[4-(2-aminoethyl)-l-pyridinio]methyl-70- [(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(l-carboxy-lmethylethoxyimino)acetamido]-3-cephem-4-carboxylic acid hydrogen sulfate (Example 1A) Compound 3-{4-[(2-aminoethyl)amino]-l-pyridinio}methyl- 7p-[(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-lmethylethoxyimino)acetamido]-3-cephem-4-carboxylate (Example 2A) P:\OPER\Kbm\specis\12628860 speci.doc Ceftazidime Test results: Table 1 Test strain Test compound MIC (jig/ml) Pseudomonas 1 aeruginosa 1 FP 1456 Ceftazidime 128

(N

0

CI

(N

V

C)

For therapeutic administration, the compound and pharmaceutically acceptable salts thereof of the present invention are used in the form of a conventional pharmaceutical preparation which contains said compound as an active ingredient, in admixture with pharmaceutically acceptable carriers such as an organic or inorganic solid or liquid excipient which is suitable for oral, parenteral or external administration. The pharmaceutical preparations may be in a solid form such as tablet, granule, powder, capsule, or in a liquid form such as solution, suspension, syrup, emulsion, lemonade and the like.

If needed, there may be included in the above preparations auxiliary substances, stabilizing agents, wetting agents and other commonly used additives such as lactose, citric acid, tartaric acid, stearic acid, magnesium stearate, terra alba, sucrose, corn starch, talc, gelatin, agar, pectin, peanut oil, olive oil, cacao butter, ethylene glycol, and the like.

While the dosage of the compound may vary from and also depend upon the age, conditions of the patient, a kind of diseases, a kind of the compound to be applied, etc.

In general amounts between 1 mg and 4,000 mg or even more per day may be administered to a patient. An average single dose of about 50 mg, 100 mg, 250 mg, 500 mg, 1000 mg or 2000 P:\OPER\Kbm\specis\12628860 specidoc Smg of the compounds of the present invention may be used in treating diseases infected by pathogenic microorganisms.

The following Preparations and Examples are given for Sthe purpose of illustrating the present invention in more detail.

(N

Preparation 1A To a solution of [(tert-butoxycarbonyl)amino]acetic acid (5.26 g) and triethylamine (3.03 g) in tetrahydrofuran 00 (120 ml) was added methyl chloroformate (2.3 ml) followed by stirring under ice-cooling for 30 minutes. To the reaction mixture was added 4-aminopyridine (2.82 and the mixture was stirred at room temperature for 16 hours. After evaporation of the solvent in vacuo, the residue was dissolved in chloroform. The solution was washed with successively saturated aqueous sodium hydrogen carbonate solution and brine. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with 5% methanol/chloroform to give tertbutyl [2-oxo-2-(4-pyridinylamino)ethyl]carbamate (4.25 g) as a solid.

1 H-NMR (CDC1 3 6 1.49 (9H, 3.94 (2H, d, J= 6.0 Hz), 5.34 (1H, brs), 7.47 (2H, d, J 6.0 Hz), 8.50 (2H, d, J Hz), 8.80 (1H, brs) Preparation 2A To a solution of tert-butyl [2-oxo-2-(4pyridinylamino)ethyl]carbamate (4.25 g) in methanol (30 ml) was added 4M hydrogen chloride solution in dioxane (15 ml) and the mixture was stirred at room temperature for 1 hour.

After evaporation of the solvent in vacuo, the residue was triturated with acetone and dried in vacuo to give 2-amino- N-(4-pyridinyl)acetamide dihydrochloride (3.34 g) as a solid.

P:\OPER\Kbm\specis\12628860 specidoc IH-NMR (DMSO-d 6 8 4.00 (2H, 8.15 (2H, d, J 6.9 Hz), 8.75 (2H, d, J= 6.9 Hz) Preparation 3A To a solution of 2-amino-N-(4-pyridinyl)acetamide 5 dihydrochloride (3.34 g) and triethylamine (4.60 g) in

(N

chloroform (120 ml) was added triphenylmethyl chloride (4.20 q and the mixture was stirred at room temperature for 2 0 hours. The reaction mixture was washed with successively aqueous citric acid solution, brine and saturated in 10 aqueous sodium hydrogen carbonate solution. The organic Slayer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The residue was triturated with diethyl ether and dried in vacuo to give N-(4-pyridinyl)-2- (tritylamino)acetamide (3.50 g) as a solid.

1 H-NMR (CDC1 3 6 2.77 (1H, t, J= 8.2 Hz), 3.23 (2H, d, J 7.8 Hz), 7.22-7.39 (15H, 7.45 (2H, d, J 6.4 Hz), 8.50 (2H, d, J 6.4 Hz), 9.23(1H, brs) Preparation 4A To a suspension of lithium aluminum hydride (1.9 g) in tetrahydrofuran (100 ml) was added N-(4-pyridinyl)-2- (tritylamino)acetamide (3.5 g) at room temperature, and the mixture was stirred under reflux for 22 hours. After cooling on ice bath, sodium fluoride (12 g) and water ml) were added to the reaction mixture. The insoluble materials were removed by filtration. The resulting filtrate was concentrated in vacuo, and the residue was dissolved in chloroform. The solution was washed with saturated aqueous sodium hydrogen carbonate solution. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The oily residue was purified by column chromatography on silica gel eluting with methanol/chloroform to give N-(4-pyridinyl)-N'-trityl- 1,2-ethanediamine (1.8 g) as an oil.

P:\OPER\Kbm\specis\12628860 specidoc 1 H-NMR (CDC1 3 6 2.44 (2H, t, J 6.0 Hz), 3.25 (2H, q, J 6.0 Hz), 4.45 (1H, brs), 6.43 (2H, d, J 6.0 Hz), 7.18-7.46 8.17 (2H, d, J 6.0 Hz) ;Z Preparation S 5 To a solution of N-(4-pyridinyl)-N'-trityl-1,2-

(N

ethanediamine (0.8 g) in chloroform (5 ml) was added di- C, tert-butyl dicarbonate (1.0 and the mixture was stirred OO under reflux for 1 hour. After evaporation of the solvent Sin vacuo, the residue was triturated with diisopropyl ether and dried in vacuo to give tert-butyl (4-pyridinyl) [2- (tritylamino)ethyl]carbamate (0.5 g) as a solid.

1 H-NMR (CDC13) 5 1.44 (9H, 2.38-2.44 (2H, 3.82-3.89 (2H, 7.15-7.41 (17H, 8.49-8.52 (2H, m) Preparation 6A To a suspension of 3,4-diaminopyridine (5.45 g) in tetrahydrofuran (75 ml) was added di-tert-butyl dicarbonate (16.35 and the mixture was stirred at room temperature for 2 hours. After evaporation of the solvent in vacuo, the residue was triturated with diisopropyl ether and dried in vacuo to give tert-butyl (3-amino-4-pyridinyl)carbamate (6.15 g) as a solid.

1 H-NMR (CDC13) 5 1.53 (9H, 3.42 (1H, brs), 6.77 (1H, brs), 7.62 (1H, d, J 5.5 Hz), 8.09 (1H, d, J 5.5 Hz), 8.13 (1H, s) Preparation 7A To a solution of tert-butyl (3-amino-4pyridinyl)carbamate (1.05 g) and [(tertbutoxycarbonyl)amino]acetic acid (1.75 g) in methylene chloride (10 ml) was added l-ethyl-3-(3dimethylaminopropyl)carbodiimide hydrochloride (1.91 and the mixture was stirred at room temperature for 18 hours.

The reaction mixture was washed with successively brine and saturated aqueous sodium hydrogen carbonate solution. The P:\OPER\Kbm\specis\12628860 specidoc Sorganic layer was dried over anhydrous sodium sulfate, Sfiltered, and concentrated in vacuo. The residue was C triturated with diisopropyl ether and dried in vacuo to give Stert-butyl [(tert-butoxycarbonyl)amino]acetyl}amino)-4- 5 pyridinyl]carbamate (1.24 g) as a solid.

(N

1 H-NMR (CDC1 3 8 1.50 (9H, 1.53 (9H, 3.97 (2H, d, J C, 5.5Hz), 5.27 (1H, brs), 7.57 (1H, brs), 7,98 (1H, d, J 00 5.5Hz), 8.22 (1H, brs), 8.38 (1H, d, J 5.5Hz), 8.38 (1H, Ss) V) 10 Preparation 8A To a solution of [(triphenylmethyl)amino]acetic acid (4.76 g) and triethylamine (1.52 g) in tetrahydrofuran ml) was added methyl chloroformate (1.15 ml) followed by stirring under ice-cooling for 30 minutes. To the reaction mixture were added a solution of trihydrochloride (3.28 g) in water (5 ml) and triethylamine (4.55 and the mixture was stirred at room temperature for 2 hours. After evaporation of the solvent in vacuo, the residue was dissolved in chloroform. The solution was washed with successively saturated aqueous sodium hydrogen carbonate solution and brine. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with 5% methanol/chloroform to give N- (5-amino-3-pyridinyl)-2-(tritylamino)acetamide (1.5 g) as an oil.

1 H-NMR (CDC13) 6 3.18 (2H, 7.21-7.43 (17H, 7.72 (1H, t, J= 2.3 Hz), 7.83 (2H, 9.15 (1H, brs) Preparation 9A To a solution of N-(5-amino-3-pyridinyl)-2- (tritylamino)acetamide (1.5 g) and triethylamine (0.37 g) in chloroform (30 ml) was added triphenylmethyl chloride (1.02 and the mixture was stirred at room temperature for 17 P:\OPER\KbM\speiS\12628860 specidoc hours. The reaction mixture was washed with successively aqueous citric acid solution, brine and saturated c aqueous sodium hydrogen carbonate solution. The organic Slayer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with 3% methanol/chloroform to give (tritylamino)-3-pyridinyl]acetamide (2.0 g) as an oil.

S1H-NMR (CDC13) 8 2.53 (1H, t, J= 8.2 Hz), 3.05 (2H, d, J= 7.8 in Hz), 5.14 (1H, 7.15 (1H, t, J 2.3 Hz), 7.21-7.35 Sm), 7.53 (1H, d, J 2.3 Hz), 7.87 (1H, d, J 2.3 Hz), 8.93 (1H, brs) Preparation To a suspension of lithium aluminum hydride (0.95 g) in tetrahydrofuran (50 ml) was added (tritylamino)-3-pyridinyl]acetamide (2.0 g) at room temperature, and the mixture was stirred under reflux for 4 hours. After cooling on ice bath, sodium fluoride (6 g) and water (5 ml) were added to the reaction mixture. The insoluble materials were removed by filtration. The resulting filtrate was concentrated in vacuo, and the residue was dissolved in chloroform. The solution was washed with 10% aqueous sodium hydroxide solution. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The oily residue was purified by column chromatography on silica gel eluting with 4% methanol/chloroform to give N-trityl-N'-[2- (0.64 g) as a solid.

1 H-NMR (CDC13) 6 2.21 (2H, t, J 5.5 Hz), 2.79 (2H, q, J 5.5 Hz), 3.65 (1H, t, J 6.0 Hz), 4.96 (1H, 5.66 (1H, t, J 2.7 Hz), 7.16-7.41 (33H, m) Preparation 11A To a solution of tert-butyl piperazine-l-carboxylate P:\OPER\Kbm\5pecis\12628860 speci.doc (1.86 g) and triethylamine (2.0 g) in ethanol (20 ml) was O added 4-chloropyridine hydrochloride (1.5 and the mixture was stirred under reflux for 4 days. The reaction Smixture was concentrated in vacuo, and the residue was dissolved in chloroform. The solution was washed with brine. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The residue Swas triturated with diisopropyl ether to give tert-butyl 4- 00

C

(4-pyridinyl)-l-piperazinecarboxylate (790 mg) as a solid.

'H-NMR (CDC13) 6 1.49 (9H, 3.13-3.33 (4H, 3.56-3.58 (4H, 6.65-6.66 (2H, 8.29-8.30 (2H, m) Preparation 12A To a solution of N-methyl-N'- (triphenylmethyl)ethylenediamine (3.80 g) and triethylamine (2.0 g) in ethanol (20 ml) was added 4-chloropyridine hydrochloride (1.5 and the mixture was stirred under reflux for 4 days. The reaction mixture was concentrated in vacuo, and the residue was dissolved in chloroform. The solution was washed with successively 10% aqueous citric acid solution, brine and saturated sodium hydrogen carbonate solution. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The oily residue was purified by column chromatography on silica gel eluting with 5% methanol/chloroform to give N-methyl-N- (4-pyridinyl)-N'-trityl-1,2-ethanediamine (1.03 g) as a solid.

1 H-NMR (CDC1 3 8 2.37-2.40 (2H, 2.97 (3H, 3.46 (2H, t, J= 6.0 Hz), 6.50 (2H, d, J 6.4 Hz), 7.16-7.27 (9H, m), 7.40-7.42 (6H, 8.20 (2H, d, J 6.4 Hz) Preparation 13A To a solution of 4-hydroxy-3-nitropyridine (3.50 g), tert-butyl (2-hydroxyethyl)carbamate (4.03 g) and triphenylphosphine (9.83 g) in tetrahydrofuran (50 ml) was P:\OPER\Kbm\speCiS\12628860 specidoc added diethyl azodicarboxylate (6.53 and the mixture was Sstirred at room temperature for 13 hours. After evaporation of the solvent in vacuo, the residue was purified by column Schromatography on silica gel eluting with 3% 5 methanol/chloroform to give tert-butyl {2-[(3-nitro-4pyridinyl)oxy]ethyl}carbamate (5.6 g) as an oil.

'1H-NMR (CDC13) 6 1.44 (9H, 3.62 (2H, q, J 5.5 Hz), 4.25 OO (2H, t, J 5.5 Hz), 7.03 (1H, d, J 6.0 Hz), 8.64 (1H, d,

(N

SJ 6.0 Hz), 9.04 (1H, s) In Preparation 14A STo a suspension of iron (1.67 g) in water (30 ml) was added concentrated hydrochloric acid (0.23 ml), and the mixture was stirred at 80 0 C for 30 minutes. To the mixture was added a solution of tert-butyl {2-[(3-nitro-4pyridinyl)oxy]ethyl}carbamate (5.6 g) in ethanol (60 ml), and the reaction mixture was stirred at 80 0 C for 30 minutes.

After cooling, the insoluble materials were removed by filtration. The filtrate was extracted with chloroform.

The combined organic layers were dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with 7% methanol/chloroform to give tert-butyl {2- [(3-amino-4-pyridinyl)oxy]ethyl}carbamate (4.84 g) as an oil.

1 H-NMR (CDC13) 6 1.45 (9H, 3.59 (2H, q, J 5.5 Hz), 3.75 (2H, brs), 4.11 (2H, t, J 5.5 Hz), 4.94 (1H, brs), 6.68 (1H, d, J 5.5 Hz), 7.96 (1H, d, J 5.5 Hz), 8.02 (1H, s) Preparation To a solution of tert-butyl {2-[(3-amino-4pyridinyl)oxy]ethyl}carbamate (2.84 g) and triethylamine (1.13 g) in chloroform (20 ml) was added triphenylmethyl chloride (3.13 and the mixture was stirred under reflux for 2 hours. The reaction mixture was washed with P:\OPER\KbM\specis\12628860 specidoc successively 10% aqueous citric acid solution, brine and O saturated aqueous sodium hydrogen carbonate solution. The organic layer was dried over anhydrous magnesium sulfate, Sfiltered, and concentrated in vacuo. The residue was 5 purified by column chromatography on silica gel eluting with 3% methanol/chloroform to give tert-butyl (tritylamino)-4-pyridinyl]oxy}ethyl)carbamate (2.40 g) as a 0 solid.

00 S1H-NMR (CDC13) 6 1.42 (9H, 3.54 (2H, q, J= 5.5 Hz), 4.13 (2H, t, J 5.5 Hz), 4.79 (1H, brs), 5.47 (1H, 6.61 (1H, Sd, J 6.0 Hz), 7.19-7.37 (16H, 7.76 (1H, d, J 6.0 Hz) Preparation 16A To a solution of 4-chloro-3-nitropyridine (3.3 g) and triethylamine (4.05 g) in ethanol (20 ml) was added tertbutyl 2-mercaptoethylcarbamate (4.25 and the mixture was stirred under reflux for 19 hours. After evaporation of the solvent in vacuo, the residue was dissolved in chloroform.

The solution was washed with successively 10% aqueous citric acid solution, brine and saturated aqueous sodium hydrogen carbonate solution. The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The residue was triturated with diisopropyl ether and dried in vacuo to give tert-butyl {2-[(3-nitro-4pyridinyl)thio]ethyl}carbamate (4.6 g) as a solid.

1 H-NMR (CDC13) 6 1.49 (9H, 3.19 (2H, t, J 7.3 Hz), 3.46 (2H, q, J 7.3 Hz), 4.95 (1H, brs), 7.59 (1H, d, J Hz), 8.63 (1H, d, J 5.5 Hz), 9.34 (1H, s) Preparation 17A To a suspension of iron (1.67 g) in water (50 ml) was added concentrated hydrochloric acid (0.35 ml), and the mixture was stirred at 80 0 C for 20 minutes. To the mixture was added a solution of tert-butyl {2-[(3-nitro-4pyridinyl)thio]ethyl}carbamate (4.6 g) in ethanol (200 ml), P:\OPER\Kbm\specis\1262886O specidoc i and the reaction mixture was stirred at 80 0 C for 1 hour.

After cooling, the insoluble materials were removed by filtration. The filtrate was extracted with chloroform.

SThe combined organic layers were dried over anhydrous 5 magnesium sulfate, filtered, and concentrated in vacuo to give tert-butyl {2-[(3-amino-4p- pyridinyl)thio]ethyl}carbamate (5.2 g) as an oil.

00 H-NMR (CDC13) 6 1.43 (9H, 3.04 (2H, t, J 6.0 Hz), 3.33 S(2H, q, J 6.0 Hz), 4.12 (2H, brs), 4.92 (1H, brs), 7.18

(N

In 10 (1H, d, J 5.0 Hz), 7.95 (1H, d, J 5.0 Hz), 8.06 (1H, s) SPreparation 18A To a solution of tert-butyl {2-[(3-amino-4pyridinyl)thio]ethyl}carbamate (2.5 g) and triethylamine g) in chloroform (30 ml) was added triphenylmethyl chloride (3.0 and the mixture was stirred at room temperature for 2 hours. The reaction mixture was washed with successively 10% aqueous citric acid solution, brine and saturated aqueous sodium hydrogen carbonate solution.

The organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with 3% methanol/chloroform to give tert-butyl (tritylamino)-4-pyridinyl]thio}ethyl)carbamate (2.3 g) as a solid.

'H-NMR (CDC13) 6 1.42 (9H, 3.05 (2H, t, J 6.0 Hz), 3.32 (2H, q, J 6.0 Hz), 4.84 (1H, brs), 6.02 (1H, 7.16 (1H, d, J 5.0 Hz), 7.21-7.35 (15H, 7.48 (1H, 7.76 (1H, d, J 5.0 Hz) Preparation 19A To a solution of (Z)-2-(5-amino-l,2,4-thiadiazol-3yl)-2-(l-tert-butoxycarbonyl-l-methylethoxyimino)acetic acid (319 g) in N,N-dimethylacetamide (1.5 L) were added potassium carbonate (113 g) and methanesulfonyl chloride P \OPER\Kbm\seciS\12628860 specidoc l (126 ml) under ice-cooling. The mixture was stirred at 10 0

C

y for 2 hours. The reaction mixture was added to a mixture of ethyl acetate and water. The organic layer was washed with water and brine to give an activated acid solution. On the S 5 other hand, a suspension of 4-methoxybenzyl 7p-amino-3chloromethyl-3-cephem-4-carboxylate hydrochloride (300 g) in C-q a mixture of water (1 L) and ethyl acetate (1 L) was 00 adjusted to pH 6 with triethylamine under ice-cooling. To

(N

Sthe resulting mixture was dropwise added the above obtained

(N

activated acid solution at 10 0 C under stirring. Stirring was continued at 5-10 0 C for 1.5 hours keeping pH of the reaction mixture at 6 with triethylamine. The organic layer was separated, washed with water and brine, and evaporated in vacuo. The concentrate was poured into diisopropyl ether (15 and the resulting precipitate was collected by filtration and dried to give 4-methoxybenzyl amino-1,2,4-thiadiazol-3-yl)-2-(l-tert-butoxycarbonyl-lmethylethoxyimino)acetamido]-3-chloromethyl-3-cephem-4carboxylate (495.7 g).

'H-NMR(DMSO-d 6 6 1.39 (9H, 1.44 (6H, 3.45-3.70 (2H, 3.76 (3H, 4.46 and 4.54 (1H, ABq, J 16 Hz), 5.10- 5.28 (2H+1H, 5.90 (1H, dd, J 4.9, 8.5 Hz), 6.94 (2H, d, J 8.7 Hz), 7.36 (2H, d, J 8.7 Hz), 8.18 (2H, brs), 9.52 (1H, d, J 8.5 Hz) Example 1A To a solution of 4-methoxybenzyl 7p-[(Z)-2-(5-amino- 1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-lmethylethoxyimino)acetamido]-3-chloromethyl-3-cephem-4carboxylate (2 g) in N,N-dimethylformamide (6 ml) was added 1,3-bis(trimethylsilyl)urea (3 g) and the mixture was stirred at room temperature for 30 minutes. To the solution was added potassium iodide (682 mg) and the mixture was stirred at room temperature for 30 minutes. To the reaction 34 P:\OPER\KbM\speCiS\1262886O specidoc mixture was added tert-butyl [2-(4-pyridinyl)ethyl]carbamate O(522 mg), and the whole mixture was stirred at room temperature for 1 hour. To the resulting reaction mixture Swas added ethyl acetate (20 ml) and the solution was washed 5 successively with water (20 ml), 10% aqueous sodium trifluoroacetate solution (10 ml) and brine (10 ml). The solution was concentrated to about 10 ml in vacuo. The Sconcentrate was poured into diisopropyl ether (100 ml) and 00 the resulting precipitate was collected by filtration and S 10 dried in vacuo. To a solution of the obtained solid in methylene chloride (7 ml) were added anisole (2.3 ml) and trifluoroacetic acid (7 ml).

The resulting solution was stirred at room temperature for 4 hours and poured into diisopropyl ether (250 ml). The resulting precipitate was collected by filtration and dried in vacuo to give a crude product (1.88 The crude product (1.88 g) was dissolved in a phosphate buffer (pH 7) and purified by preparative high-performance liquid chromatography (HPLC) utilizing ODS column. The eluate containing a desired product was concentrated to about 30 ml in vacuo.

The concentrate was adjusted to about pH 3 by addition of concentrated hydrochloric acid and chromatographed on Diaion (registered trademark) HP-20 (Mitsubishi Chemical Corporation) eluting with 10% aqueous 2-propanol. The eluate was concentrated to about 1 L in vacuo, added with 2M aqueous sulfuric acid solution (0.44 ml) and lyophilized to give 3-[4-(2-aminoethyl)-l-pyridinio]methyl-7p-[(Z)-2-(5amino-1,2,4-thiadiazol-3-yl)-2-(l-carboxy-lmethylethoxyimino)acetamido]-3-cephem-4-carboxylic acid hydrogen sulfate (610 mg) as an amorphous solid.

1 H-NMR (D 2 0) 8 1.58 (6H, 3.17 and 3.62 (2H, ABq, J 18.0 Hz), 3.20-3.57 (4H, 5.28 and 5.52 (2H, ABq, J 15.0 P:\OPER\Kbm\specis\12628860 specidoc Hz), 5.28 (1H, d, J 5.0 Hz), 5.90 (1H, d, J 5.0 Hz), S8.01 (2H, d, J 7.0 Hz), 8.90 (2H, d, J 7.0 Hz) Example 2A STo a solution of 7p-[(Z)-2-(5-amino-l,2,4-thiadiazol- 3-yl)-2-(l-carboxy-l-methylethoxyimino)acetamido]-3chloromethyl-3-cephem-4-carboxylic acid trifluoroacetate Ci (750 mg), N-(4-pyridinyl)-N'-trityl-l,2-ethanediamine (800 Smg) in N,N-dimethylformamide (7 ml) was added diisopropylethylamine (0.76 ml), and the mixture was stirred qq 10 at room temperature for 3 hours. The reaction mixture was Spoured into ethyl acetate (120 ml), and the resulting precipitate was collected by filtration and dried in vacuo to give a crude product (1.1 To a solution of the crude product in methylene chloride (3.0 ml) were added anisole (1.0 ml) and trifluoroacetic acid (2.0 ml), and the mixture was stirred at room temperature for 3 hours. The reaction mixture was poured into diisopropyl ether (120 ml). The resulting precipitate was collected by filtration and dried in vacuo to give a crude product (1.3 g) which was purified by preparative high-performance liquid chromatography (HPLC) utilizing ODS column. The eluate containing a desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to about pH 3 by addition of concentrated hydrochloric acid and chromatographed on Diaion (registered trademark) HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo and lyophilized to give 3-{4-[(2-aminoethyl)amino]-1pyridinio}methyl-7p-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)- 2-(1-carboxy-l-methylethoxyimino)acetamido]-3-cephem-4carboxylate (49.0 mg) as an amorphous solid.

1 H-NMR (D 2 0) 6 1.50 (6H, 3.16 (1H, d, J 17.9 Hz), 3.27 (2H, t, J 6.2 Hz), 3.58 (1H, d, J 17.9 Hz), 3.70 (2H, t, P:\OPER\KbM\speCiS\1262886D specidoc J 6.2 Hz), 4.84 (1H, d, J 14.7 Hz), 5.08 (1H, d, J 14. 7 Hz) 5. 24 (1H, d, J 4. 6 Hz) 5. 82 (1H, d, J 4. 6 Hz), 6.90 (2H, d, J 6.4 Hz), 8.10 (1H, brs), 8.25 (1H, ;Z brs) Example 3A F(2-Aminoethyl)amino] -l-pyridinio~methyl-7p- 2-(5-amino-1,2,4-thiadiazol-3-yl)-2-(1-carboxy-lmethylethoxyimino)acetamido] -3-cephem-4-carboxylate The title compound was obtained in the same manner as in Example 2A using tert-butyl (4-pyridinyl) [2- (tritylamino) ethyl] carbamate.

1 H-NMR (D 2 0) 8 1. 50 (6H, s) 3. 16 (1H, d, J 17. 9 Hz) 3.27 (2H, t, J 6.2 Hz), 3.58 (1H, d, J 17.9 Hz), 3.70 (2H, t, J =6.2 Hz), 4.84 (1H, d, J 14.7 Hz), 5.08 (1H, d, J 14. 7 Hz) 5. 24 (1H, d, J 4. 6 Hz) 5. 82 (1H, d, J 4. 6 Hz), 6.90 (2H, d, J 6.4 Hz), 8.10 (1H, brs), 8.25 (1H, brs) Example 4A 3- {4-Amino-3- [(aminoacetyl) amino] -1-pyridinio~methyl- 7p-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(l-carboxy-lmethylethoxyimino) acetamido] -3-cephem-4-carboxylate The title compound was obtained in the same manner as in Example 2A using tert-butyl butoxycarbonyl) amino] acetyl} amino) -4 -pyridinyllcarbamate.

'H-NMR (D 2 0) 5 1.49 (3H, s) 1. 50 (3H, s) 3.16 (1H, d, J= 17.9 Hz), 3.60 (1H, d, J 17.9 Hz), 4.07 (2H, 4.84 (1H, d, J 14.7 Hz), 5.12 (1H, d, J 14.7 Hz), 5.26 (1H, d, J 4.8 Hz), 5.83 (1H, d, J 4.8 Hz), 7.01 (1H, d, J 7.1 Hz), 8.12 dd, J 7.1, 0.9 Hz), 8.43 (1H, d, J 0.9 Hz) Example 3-{3-Amino-5- [(2-aminoethyl)amino] -1-pyridinio~methyl- 73- (5-amino-1,2,4-thiadiazol-3-yl) (1-carboxy-1methylethoxyimino) acetamido] -3-cephem-4-carboxylate P:\OPER\Kbni\speciS\12628860 specidoc tetrahydrochioride The title compound was obtained in the same manner as in Example 2A using N-trityl-N' (tritylamino) ethyl] ;Z pyridinediamine.

'H-NMR (D 2 0) 8 1.60 (6H, s) 3.21 (2H, d, J 6. 0 Hz) 3.23 (1H, t, J 17.9 Hz), 3.51 (2H, t, J 6.0 Hz), 3.62 (1H, d, J 17.9 Hz), 5.03 (1H, d, J 14.7 Hz), 5.28 (1H, d, J= 00 4.6 Hz), 5.29 (1H, d, J 14.7 Hz), 5.86 (1H, d, J 4.6

(N

Hz) 6.86 (1H, s) 7.57 (1H, s) 7.61 (1H, s) Example 6A 7f-[(Z)-2-(5-Amino-1,2,4-thiadiazol-3-yl)-2-(lcarboxy-l-methylethoxyimino) acetamido] (1-piperazinyl) 1 -pyridinio] methyl cephem-4 -carboxylate The title compound was obtained in the same manner as in Example 2A using tert-butyl 4-(4-pyridinyl)-lpiperazinecarboxylate.

1 H-NMR (D 2 0) 8 1.50 (3H, s) 1. 50 (3H, s) 3.17 (1H, d, J= 17.9 Hz), 3.45 (4H, t, J =5.5 Hz), 3.60 (1H, d, J 17.9 Hz), 3.97 (4H, t, J 5.5 Hz), 4.90 (1H, d, J 14.7 Hz), 5.13 (1H, d, -J 14.7 Hz), 5.26 (1H, d, J 5. 0 Hz) 5.84 (1H, d, J 5. 0 Hz) 7.14 (2H, d, J 7.6 Hz) 8.28 (2H, d, J 7.6 Hz) Example 7A [(2-Aminoethyl) (methyl)amino] -l-pyridinio~methyl- 7p-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(l-carboxy-lmethylethoxyimino) acetamido] -3 -cephem-4 -carboxylate The title compound was obtained in the same manner as in Example 2A using N-methyl-N- (4-pyridinyl) -trityl-l,2ethanediamine.

1 H-NMR (D 2 0) 8 1. 59 (6H, s) 3.22 (1H, d, J 17. 9 Hz) 3.24 (3H, 3.32 (2H, t, J 6.4 Hz), 3.60 (1H, d, J 17.9 Hz), 3.93 (2H, t, J 6.4 Hz), 4.97 (1H, d, J 14.7 Hz), 13 (1H, d, J 14. 7 Hz) 5. 27 (1H, d, J 4. 8 Hz) 5.8 7 P:\OPER\Kbm\specis\12628860 specidoc (1H, d, J 4.8 Hz), 7.02 (2H, brs), 7.33 (2H, d, J 7.3 Hz) Example 8A 13-[3-Amino-4-(2-aminoethoxy)-1-pyridinio]methyl-7p- S 5 [(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(l-carboxy-lmethylethoxyimino)acetamido]-3-cephem-4-carboxylate C- The title compound was obtained in the same manner as O0 in Example 2A using tert-butyl (2-{[3-(tritylamino)-4- Spyridinyl]oxy}ethyl)carbamate.

kI 10 H-NMR (D 2 0) 8 1.49 (3H, 1.49 (3H, 3.14 (1H, d, J S17.9 Hz), 3.57 (2H, t, J 4.8 Hz), 3.58 (1H, d, J 17.9 Hz), 4.58 (2H, t, J 4.8 Hz), 4.99 (1H, d, J 14.2 Hz), 5.23 (1H, d, J 4.6 Hz), 5.28 (1H, d, J 14.2 Hz), 5.83 (1H, d, J 4.6 Hz), 7.29 (1H, d, J 6.9 Hz), 8.17 (1H, d, J 6.9 Hz), 8.17 (1H, s) Example 9A 3-{3-Amino-4-[(2-aminoethyl)thio]-l-pyridinio}methyl- 7p-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(l-carboxy-lmethylethoxyimino)acetamido]-3-cephem-4-carboxylate The title compound was obtained in the same manner as in Example 2A using tert-butyl (2-{[3-(tritylamino)-4pyridinyl]thio}ethyl)carbamate.

1 H-NMR (D 2 0) 8 1.58 (6H, 3.19 (1H, d, J 17.9 Hz), 3.38 (2H, t, J 6.9 Hz), 3.60 (2H, t, J 6.9 Hz), 3.61 (1H, d, J 17.9 Hz), 5.12 (1H, d, J 14.2 Hz), 5.27 (1H, d, J 5.1 Hz), 5.32 (1H, dd, J 14.2, 3.7 Hz), 5.87 (1H, d, J 5.1 Hz), 7.67 (1H, d, J 6.4 Hz), 8.10 (1H, d, J 6.4 Hz), 8.12 (1H, s) Another type of the cephem compound of the present invention is explained in the following.

The cephem compounds of the present invention are novel and can be represented by the following general formula P:\OPER\Kbm\specis\12628860 speci.dOC 1 2 in R R 0 X 3 0D 0 R §N 8 I S R C- CONHOH

R

4 s R 5

-CH

2

[I]

COO 1 17 eC R R O0 wherein Q R and R 2 are each hydrogen, lower alkyl, qn hydroxy(lower)alkyl, protected hydroxy(lower)alkyl or halo(lower)alkyl, or

R

1 and R 2 are bonded together and form lower alkylene;

R

3 is carboxy or protected carboxy;

R

4 is amino or protected amino;

R

5 is hydrogen, halogen or lower alkyl;

R

6 is lower alkyl which may have suitable substituent(s),

R

7 is hydrogen or amino protecting group, or

R

6 and R 7 are bonded together and form lower alkylene;

R

8 is amino or protected amino; X is -NHCO-, -NHCONH- or -N(R 9 (wherein R 9 is hydrogen, lower alkyl or amino protecting group); A is lower alkylene; and n is 0 or 1.

As to the compound the following points are to be noted.

That is, the compound includes syn isomer (Z form), anti isomer (E form) and a mixture thereof. Syn isomer (Z form) means one geometrical isomer having the partial structure represented by the following formula: P:\OPER\Kbm\speci9\12628860 specidoc 1 2 N R R 2 O xR3 N-0 R

II

4N-C-CO- C-I wherein R 1

R

2

R

3

R

4 and R 5 are each as defined above, and anti isomer (E form) means the other geometrical isomer having the partial structure represented by the following 00 0- 5 formula: 2 1 SR R l n 3 x 0 R O-N

II

C4 N s c-co- R S R wherein R 1

R

2

R

3

R

4 and R 5 are each as defined above, and all of such geometrical isomers and mixtures thereof are included within the scope of this invention.

In the present specification and claims, the partial structure of these geometrical isomers and mixtures thereof are represented for convenience' sake by the following formula: 1 2 R R OXR3

N

II

N~ C-CO- R S R wherein R 1

R

2

R

3

R

4 and R 5 are each as defined above.

Another point to be noted is that the pyrazolio moiety of the compound can also exist in the tautomeric form, and such tautomeric equilibrium can be represented by the following formula.

P:\OPER\Kbm\specis\12628860 specidoc in -A-R 8 R N N H N H N N N N I6 I7 I I7 R R R R

(B)

wherein R 6

R

7

R

8 X, A and n are each as defined above.

CI Both of the above tautomeric isomers are included 00 within the scope of the present invention, and in the 0 5 present specification and claims, however, the compound [I]

(N

in is represented for convenience' sake by one expression of 0 the pyrazolio group of the formula The cephem compound of the present invention can be prepared by the following processes as illustrated in the following.

P:\OPER\Kbm\specis\1262886G specidoc Process 1 (X)--R8 HN r

CH-N

0 2 N N .COG1 6 1 7 R R

N

N -CCOOH RS R

[III

or its reactive derivative at the amino group, or a salt thereof

[III]

or its reactive derivative at the carboxy group, or a salt thereof k ;00 (9

IN

,6 R 7 or a salt thereof P:\OPER\Kbm\speci9\12628860 specidoc Process 2 R1 R2 0xR N 8 a 11 S n N C-CONHfl

R

4 R rHN N S0 co®N

R

6

R

7 [Ta] or a salt thereof Elimination reaction of the Iamino protecting group

(X)-A-N*H

2

N

1

I®E

coo IIb or a salt thereof P:\OPER\Kbm\specjS\12628860 specidoc Process 3 N \N

-H

1 6 1 7 R R lIV] or a salt thereof

IV]

or a salt thereof (i) 8

ZE

N

1 7

R

[VI]

or a salt thereof (ii) I E coo 1-I-H

N

1 7

R

or a salt thereof 1 2 3 4 5 6 7 8 wherein R1, R R R R X, A and n are each as defined above, R 8a is protected amino,

R

10 is protected carboxy, P:\OPER\Kbm\speciS\12628860 specidoc Y is a leaving group, and ZO is an anion.

;Z The starting compounds [III and [IV] can be prepared by the following processes.

00 P:\OPER\Kbm\specis\12628860 specidoc Process A .4N

CH

2

Y

0 R 12

[VII]

or a salt thereof -A-R 8 n N R1 N N 16 17oI

R

6

R

7

[VIII]

(i) salt thereof R 11

[IX]

or a salt thereof s WX -A-R 8 HCH 2

N

0 N N coo®E 16 17

[RIR

or a salt thereof P:\OPER\Kbm\specis\12628860 specidoc Process B 1 2 R R 0 R 00

[XI]

VB or its reactive or its reactive derivative at the derivative at the C( amino group, carboxy group, or a salt thereof or a salt thereof 1 2 R R x< 3 N-

N

II

S

N (C-CONH 4,R S 5 N CH2-Y R S R 1 0

[IV]

or a salt thereof wherein R 1

R

2

R

3

R

4

R

s

R

6 R R 8 R 1 X, A, n, Y and ZO are each as defined above,

R

1 is protected amino,

R

12 is protected carboxy, and

R

13 is amino protecting group.

The starting compounds and [VIII] or salts thereof can be prepared by the methods disclosed in the Preparations 1B-12B described later or similar manners thereto. The P: \OPER\Kbm\speci8\12628860 specidoc starting compounds and [VIII] or salts thereof can be Salso prepared by the methods disclosed in W002/090364.

C In the above and subsequent descriptions of this Sspecification, suitable examples of the various definitions are explained in detail as follows.

The term "lower" is used to mean a group having 1 to 6, preferably 1 to 4, carbon atom(s), unless otherwise indicated.

00 C Suitable "halogen" includes chlorine, bromine, iodine S 10 and fluorine.

Suitable "lower alkyl" and "lower alkyl" moiety in "hydroxy(lower)alkyl", "protected hydroxy(lower)alkyl", "halo(lower)alkyl", "aryl(lower)alkyl", "aryl(lower)alkyloxy" and "lower alkylthio" include straight or branched alkyl having 1 to 6 carbon atom(s), such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, secbutyl, tert-butyl, pentyl, tert-pentyl and hexyl, in which more preferred one is CI-C 4 alkyl.

Suitable examples of "substituent(s)" in "lower alkyl which may have suitable substituent(s)" may include one or more (preferably 1 to 3) substituent(s) selected from amino, hydroxy, halogen, cyano, lower alkoxy, lower alkylthio, aryl(lower)alkyloxy, carboxy, and the like.

Suitable "hydroxy(lower)alkyl" includes hydroxy(C 1 Cs)alkyl such as hydroxymethyl, l-hydroxyethyl, 2hydroxyethyl, 1-hydroxypropyl, 2-hydroxypropyl, 3hydroxypropyl, 4-hydroxybutyl, 5-hydroxypentyl and 6hydroxyhexyl, in which more preferred one is hydroxy(C 1

C

4 alkyl.

Suitable "halo(lower)alkyl" includes straight or branched alkyl having 1 to 6 carbon atoms substituted by 1 to 5 halogen atoms such as chlorine, bromine, iodine and fluorine. Preferred examples of "halo(lower)alkyl" include P:\OPER\Kb\specis\12628860 specidoc fluoromethyl, difluoromethyl, trifluoromethyl, chloromethyl, Sbromomethyl, 2-fluoroethyl, 2,2-difluoroethyl, 2,2,2- C trifluoroethyl, 2-chloroethyl, 2,2-dichloroethyl, 2,2,2- Strichloroethyl, 3-fluoropropyl and 2,2,3,3,3pentafluoropropyl, in which more preferred one is halo(C 1

C

4 )alkyl.

Suitable "lower alkylene" at A includes straight or Sbranched alkylene having 1 to 6 carbon atoms, such as 00 C methylene, ethylene, trimethylene, tetramethylene, S 10 pentamethylene, hexamethylene and propylene, in which more Spreferred one is straight alkylene having 1 to 3 carbon atoms.

Suitable "lower alkylene" formed by R 1 and R 2 includes straight or branched alkylene having 1 to 6, preferably 2 to 4 carbon atoms, such as methylene, ethylene, trimethylene and tetramethylene, in which more preferred one is straight alkylene having 2 or 3 carbon atoms.

Suitable "lower alkylene" formed by R 6 and R 7 includes straight alkylene having 1 to 6, preferably 2 to 4 carbon atoms, such as methylene, ethylene, trimethylene and tetramethylene, in which more preferred one is straight alkylene having 2 or 3 carbon atoms.

Suitable "aryl" and "aryl" moiety in "aryl(lower)alkyl" and "aryl(lower)alkyloxy" includes C 6 -C12 aryl such as phenyl and naphthyl, in which more preferred one is phenyl.

Suitable "aryl(lower)alkyl" includes mono-, di- or triphenyl(lower)alkyl such as benzyl, phenethyl, benzhydryl and trityl.

Suitable "lower alkanoyl" and "lower alkanoyl" moiety in "lower alkanoylamino" and "lower alkanoyloxy" include straight or branched alkanoyl having 1 to 6 carbon atoms, such as formyl, acetyl, propionyl, butyryl, isobutyryl, P;\OPER\KbM\speCiS\12628860 specidoc valeryl, isovaleryl, pivaloyl and hexanoyl, in which more preferred one is C 1

-C

4 alkanoyl.

Suitable "lower alkoxy" and "lower alkoxy" moiety in "lower alkoxycarbonyl", "lower alkoxycarbonylamino" and "lower alkoxycarbonyloxy" includes straight or branched alkoxy having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tertbutoxy, pentyloxy, tert-pentyloxy and hexyloxy, in which 00 more preferred one is C 1

-C

4 alkoxy.

Suitable "amino protecting group" in "protected amino" includes an acyl group as mentioned below, substituted or unsubstituted aryl(lower)alkylidene benzylidene, hydroxybenzylidene, etc.], aryl(lower)alkyl such as mono-, di- or triphenyl(lower)alkyl benzyl, phenethyl, benzhydryl, trityl, etc.], and the like.

Suitable "acyl" includes lower alkanoyl formyl, acetyl, propionyl, hexanoyl, pivaloyl, etc.], mono(or di or tri)halo(lower)alkanoyl chloroacetyl, trifluoroacetyl, etc.], lower alkoxycarbonyl methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, tertpentyloxycarbonyl, hexyloxycarbonyl, etc.], carbamoyl, aroyl benzoyl, toluoyl, naphthoyl, etc.], aryl(lower)alkanoyl phenylacetyl, phenylpropionyl, etc.], aryloxycarbonyl phenoxycarbonyl, naphthyloxycarbonyl, etc.], aryloxy(lower)alkanoyl phenoxyacetyl, phenoxypropionyl, etc.], arylglyoxyloyl phenylglyoxyloyl, naphthylglyoxyloyl, etc.], aryl(lower)alkoxycarbonyl which optionally substituted by suitable substituent(s) benzyloxycarbonyl, phenethyloxycarbonyl, p-nitrobenzyloxycarbonyl, etc.], and the like.

Preferable examples of "amino protecting group" include aryl(lower)alkyl and acyl, in which more preferred P:\OPER\Kbm\specis\12628B6D specidoc ones are aryl(lower)alkyl, lower alkanoyl and lower alkoxycarbonyl, and particularly preferred ones are mono-, di- or triphenyl(C 1 -Cs)alkyl, C1-C 6 alkanoyl and (C 1

C

6 )alkoxycarbonyl.

Preferable examples of "protected amino" include aryl(lower)alkylamino and acylamino, in which more preferred ones are aryl(lower)alkylamino, lower alkanoylamino and lower alkoxycarbonylamino, and particularly preferred ones 00 are mono-, di- or triphenyl(C 1 -C)alkylamino, C 1

-C

6 alkanoylamino and (C 1 -C)alkoxycarbonylamino.

Suitable "protected hydroxy" in the "protected hydroxy(lower)alkyl" includes acyloxy group, aryl(lower)alkyloxy group, and the like. Suitable "acyl" moiety in the "acyloxy" includes lower alkanoyl formyl, acetyl, propionyl, hexanoyl, pivaloyl, etc.], mono(or di or tri)halo(lower)alkanoyl, chloroacetyl, trifluoroacetyl, etc.], lower alkoxycarbonyl, methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, tertpentyloxycarbonyl, hexyloxycarbonyl, etc.], carbamoyl, and the like. Suitable "aryl(lower)alkyl" moiety in the "aryl(lower)alkyloxy" includes mono-, di- or triphenyl(lower)alkyl benzyl, phenethyl, benzhydryl, trityl, etc.], and the like.

Suitable "protected carboxy" includes esterified carboxy and the like, and concrete examples of esterified carboxy include lower alkoxycarbonyl methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, tert-butoxycarbonyl, pentyloxycarbonyl, hexyloxycarbonyl, 1cyclopropylethoxycarbonyl, etc.] which may have suitable substituent(s), for example, lower alkanoyloxy(lower)alkoxycarbonyl acetoxymethoxycarbonyl, propionyloxymethoxycarbonyl, P:\OPER\Kbm\BpeCi9\12628860 specidoc butyryloxymethoxycarbonyl, valeryloxymethoxycarbonyl, pivaloyloxymethoxycarbonyl, 1-acetoxyethoxycarbonyl, 1propionyloxyethoxycarbonyl, 2-propionyloxyethoxycarbonyl, hexanoyloxymethoxycarbonyl, etc.], lower alkanesulfonyl (lower) alkoxycarbonyl, 2mesylethoxycarbonyl, etc.] or mono(or di or tri)halo(lower)alkoxycarbonyl 2-iodoethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, etc.]; lower 00 alkenyloxycarbonyl vinyloxycarbonyl, allyloxycarbonyl, etc.]; lower alkynyloxycarbonyl ethynyloxycarbonyl, propynyloxycarbonyl, etc.]; aryl(lower)alkoxycarbonyl which may have suitable substituent benzyloxycarbonyl, 4methoxybenzyloxycarbonyl, 4-nitrobezyloxycarbonyl, phenethyloxycarbonyl, trityloxycarbonyl, benzhydryloxycarbonyl, bis(methoxyphenyl)methoxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 4-hydroxy-3,5-di-tertbutylbenzyloxycarbonyl, etc.]; aryloxycarbonyl which may have suitable substituent(s) phenoxycarbonyl, 4chlorophenoxycarbonyl, tolyloxycarbonyl, 4-tertbutylphenoxycarbonyl, xylyloxycarbonyl, mesityloxycarbonyl, cumenyloxycarbonyl, etc.]; and the like.

Preferable examples of "protected carboxy" include lower alkoxycarbonyl and aryl(lower)alkoxycarbonyl which may have suitable substituent(s), in which more preferred one is

(C

1

-C

6 alkoxycarbonyl.

Suitable "leaving group" includes halogen chlorine, bromine, iodine, etc.] or acyloxy such as arylsulfonyloxy benzenesulfonyloxy, tosyloxy, etc.], lower alkylsulfonyloxy mesyloxy, etc.], lower alkanoyloxy acetyloxy, propionyloxy, etc.], and the like.

Suitable "anion" includes formate, acetate, P:\OPER\Kbm\specis\1262886O specidoc trifluoroacetate, maleate, tartrate, methanesulfonate, Sbenzenesulfonate, toluenesulfonate, chloride, bromide, iodide, sulfate, phosphate, and the like.

SSuitable pharmaceutically acceptable salts of the compound are conventional non-toxic salts and include, for example, a salt with a base or an acid addition salt such as a salt with an inorganic base, for example, an Salkali metal salt sodium salt, potassium salt, etc.], an alkaline earth metal salt calcium salt, magnesium S 10 salt, etc.], an ammonium salt; a salt with an organic base, Sfor example, an organic amine salt trimethylamine C salt, triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt, etc.]; an inorganic acid addition salt hydrochloride, hydrobromide, sulfate, hydrogensulfate, phosphate, etc.]; an organic carboxylic or sulfonic acid addition salt formate, acetate, trifluoroacetate, maleate, tartrate, citrate, fumarate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc.]; and a salt with a basic or acidic amino acid arginine, aspartic acid, glutamic acid, etc.].

The preferred embodiment of the cephem compound of the present invention represented by the general formula is as follows.

The compound of the formula wherein

R

1 and R 2 are each hydrogen, lower alkyl, hydroxy(lower)alkyl, acyloxy, aryl(lower)alkyloxy or halo(lower)alkyl, or

R

1 and R 2 are bonded together and form lower alkylene;

R

3 is carboxy or esterified carboxy;

R

4 is amino, aryl(lower)alkylamino or acylamino;

R

6 is lower alkyl which may have suitable substituent(s); P:\OPER\Kbm\speCis\12628860 specidoc

R

7 is hydrogen, aryl(lower)alkyl or acyl, or

SR

6 and R 7 are bonded together and form lower alkylene; C

R

8 is amino, aryl(lower)alkylamino or acylamino; and SX is -NHCO-, -NHCONH- or -N(R 9 (wherein R 9 is hydrogen, lower alkyl, aryl(lower)alkyl or acyl), or a pharmaceutically acceptable salt thereof.

The compound of above wherein SR and R 2 are each hydrogen, lower alkyl, 00 hydroxy(lower)alkyl, lower alkanoyloxy, lower alkoxycarbonyloxy, aryl(lower)alkyloxy or Shalo(lower)alkyl, or

C

R

1 and R 2 are bonded together and form lower alkylene;

R

3 is carboxy or lower alkoxycarbonyl;

R

4 is amino, aryl(lower)alkylamino, lower alkanoylamino or lower alkoxycarbonylamino;

R

6 is lower alkyl which may have suitable substituent(s);

R

7 is hydrogen, aryl(lower)alkyl, lower alkanoyl or lower alkoxycarbonyl, or

R

6 and R 7 are bonded together and form lower alkylene;

R

8 is amino, aryl(lower)alkylamino, lower alkanoylamino or lower alkoxycarbonylamino; and X is -NHCO-, -NHCONH- or -N(R 9 (wherein R 9 is hydrogen, lower alkyl, aryl(lower)alkyl, lower alkanoyl or lower alkoxycarbonyl), or a pharmaceutically acceptable salt thereof.

The compound of above wherein

R

1 and R 2 are each hydrogen, Ci-C 6 alkyl, hydroxy(C 1

-C

6 )alkyl, C1-C 6 alkanoyloxy, (C 1

-C

6 alkoxycarbonyloxy,

(C

6

-C

12 )aryl(C 1

-C

6 )alkyloxy or halo(Cl-C 6 )alkyl, or

R

1 and R 2 are bonded together and form Ci-C 6 alkylene;

R

3 is carboxy or (Ci-C 6 )alkoxycarbonyl;

R

4 is amino, mono-, di- or triphenyl(Ci-C) alkylamino, Ci-C 6 alkanoylamino or (Ci-C) alkoxycarbonylamino; P:\OPER\Kbm\specis\12628860 specidoc

R

5 is hydrogen, halogen or Ci-C 6 alkyl;

SR

6 is Ci-Cs alkyl which may have suitable substituent(s) selected from hydroxy, halogen, cyano, Ci-C 6 alkoxy, SCi-C 6 alkylthio, (C6-C 1 2 )aryl(C 1

-C

6 )alkyloxy and carboxy;

R

7 is hydrogen, mono-, di- or triphenyl(C 1 -C6)alkyl, Ci-Cs alkanoyl or (C 1

-C

6 )alkoxycarbonyl, or

R

6 and R 7 are bonded together and form Ci-Cs alkylene;

SR

8 is amino, mono-, di- or triphenyl(Ci-C6)alkylamino, Ci-Cs 00 alkanoylamino or (C 1 -Cs)alkoxycarbonylamino; and X is -NHCO-, -NHCONH- or -N(R 9 (wherein R 9 is hydrogen, Slower alkyl, mono-, di- or triphenyl(C 1 -Cs)alkyl, Ci-C6 alkanoyl or (Ci-Cs) alkoxycarbonyl) A is C 1

-C

6 alkylene; or a pharmaceutically acceptable salt thereof.

The compound of the formula wherein R 1 and R 2 are each hydrogen, lower alkyl, hydroxy(lower)alkyl or halo(lower)alkyl, or R 1 and R 2 are bonded together and form lower alkylene, or a pharmaceutically acceptable salt thereof.

The compound of the formula wherein R 6 is lower alkyl, and R 7 is hydrogen, or a pharmaceutically acceptable salt thereof.

The compound of the formula wherein X is -NHCO- or -NHCONH-, or a pharmaceutically acceptable salt thereof.

The compound of the formula wherein R 3 is carboxy,

R

4 is amino, and R 8 is amino, or a pharmaceutically acceptable salt thereof.

The processes for preparing the compound of the present invention are explained in detail in the following.

Process 1 The compound or a salt thereof can be prepared by reacting the compound [II] or its reactive derivative at the amino group, or a salt thereof with the compound [III] or P:\OPER\Kbm\specis\12628860 specidoc its reactive derivative at the carboxy group, or a salt thereof.

c Suitable reactive derivative at the amino group of the Scompound [II] includes Schiff's base type imino or its tautomeric enamine type isomer formed by the reaction of the compound [II] with a carbonyl compound such as aldehyde, ketone and the like; a silyl derivative formed by the Sreaction of the compound [II] with a silyl compound such as 00 c bis(trimethylsilyl)acetamide, mono(trimethylsilyl)acetamide N-(trimethylsilyl)acetamide], bis(trimethylsilyl)urea Sand the like; a derivative formed by the reaction of the C compound [II] with phosphorus trichloride or phosgene.

Suitable salts of the compound [II] and its reactive derivative can be referred to the ones as exemplified for the compound Suitable reactive derivative at the carboxy group of the compound [III] includes an acid halide, an acid anhydride, an activated amide, and an activated ester. A suitable example of the reactive derivatives may be an acid chloride; an acid azide; a mixed acid anhydride with an acid such as substituted phosphoric acid dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoric acid, halogenated phosphoric acid, etc.], dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, sulfuric acid, alkanesulfonic acid methanesulfonic acid, etc.], aliphatic carboxylic acid acetic acid, propionic acid, butyric acid, isobutyric acid, pivalic acid, pentanoic acid, isopentanoic acid, 2-ethylbutyric acid, trichloroacetic acid, etc.] and aromatic carboxylic acid benzoic acid, etc.]; a symmetrical acid anhydride; an activated amide with imidazole, 4-substituted imidazole, dimethylpyrazole, triazole or tetrazole; an activated ester cyanomethyl ester, methoxymethyl ester, P:\OPER\Kbm\specis\12628860 specidoc dimethyliminomethyl [(CH 3 2 ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenyl thioester, p-nitrophenyl thioester, p-cresyl thioester, carboxymethyl thioester, pyranyl ester, pyridyl ester, piperidyl ester, 8quinolyl thioester, etc.]; or an ester with an N-hydroxy compound N,N-dimethylhydroxylamine, l-hydroxy-2-(1H)pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide, Nhydroxy-1H-benzotriazole, etc.]. These reactive derivatives can optionally be selected from them according to the kind of the compound [III] to be used.

Suitable salts of the compound [III] and its reactive derivative can be referred to the ones as exemplified for the compound The reaction is usually carried out in a conventional solvent such as water, alcohol methanol, ethanol, etc.], acetone, dioxane, acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine or any other organic solvent which does not adversely affect the reaction. These conventional solvents may also be used in a mixture with water.

In this reaction, when the compound [III] is used in free acid form or its salt form, the reaction is preferably carried out in the presence of a conventional condensing agent such as N,N'-dicyclohexylcarbodiimide; N-cyclohexyl- N'-morpholinoethylcarbodiimide; N-cyclohexyl-N'-(4diethylaminocyclohexyl)carbodiimide; N,N'diethylcarbodiimide; N,N' -diisopropylcarbodiimide; N-ethyl- N'-(3-dimethylaminopropyl)carbodiimide; N,N'-carbonyl-bis- (2-methylimidazole); pentamethyleneketene-N-cyclohexylimine; diphenylketene-N-cyclohexylimine; ethoxyacetylene; 1-alkoxy- P:\OPER\KbM\speCiS\12628860 specidoc 1-chloroethylene; trialkyl phosphite; ethyl polyphosphate; isopropyl polyphosphate; phosphorus oxychloride (phosphoryl chloride); phosphorus trichloride; thionyl chloride; oxalyl Schloride; lower alkyl haloformate ethyl chloroformate, isopropyl chloroformate, etc.], triphenylphosphine; 2-ethyl-7-hydroxybenzisoxazolium salt; hydroxide intramolecular salt; 1-(p-chlorobenzenesulfonyloxy)-6- 00 c chloro-lH-benzotriazole; so-called Vilsmeier reagent S 10 prepared by the reaction of N,N-dimethylformamide with Sthionyl chloride, phosgene, trichloromethyl chloroformate, phosphorus oxychloride, etc.; and the like.

The reaction may also be carried out in the presence of an inorganic or organic base such as an alkali metal bicarbonate, tri(lower)alkylamine, pyridine, N- (lower)alkylmorpholine, N,N-di(lower)alkylbenzylamine, and the like.

The reaction temperature is not critical, and the reaction is usually carried out under cooling to warming.

Process 2 The compound [Ib] or a salt thereof can be prepared by subjecting the compound [Ia] or a salt thereof to elimination reaction of the amino protecting group.

Elimination reaction is carried out in accordance with a conventional method such as hydrolysis and the like.

The hydrolysis is preferably carried out in the presence of a base or an acid including Lewis acid.

Suitable base includes an inorganic base and an organic base such as an alkali metal sodium, potassium, etc.], an alkaline earth metal magnesium, calcium, etc.], the hydroxide or carbonate or hydrogencarbonate thereof, trialkylamine trimethylamine, triethylamine, etc.], picoline, P:\OPER\Kbm\speCis\12628860 specidoc diazabicyclo[4.3.0]non-5-ene, 1,4-diazabicyclo[2.2.2]octane, 1,8-diazabicyclo[5.4.0]undec-7-ene, and the like.

C Suitable acid includes an organic acid formic Sacid, acetic acid, propionic acid, trichloroacetic acid, trifluoroacetic acid, etc.], and an inorganic acid hydrochloric acid, hydrobromic acid, sulfuric acid, hydrogen chloride, hydrogen bromide, etc.].

SThe elimination using Lewis acid such as trihaloacetic 00 c acid trichloroacetic acid, trifluoroacetic acid, S 10 etc.], and the like is preferably carried out in the Spresence of cation trapping agents anisole, phenol, etc.].

The reaction is usually carried out in a solvent such as water, alcohol methanol, ethanol, etc.], methylene chloride, tetrahydrofuran, a mixture thereof or any other solvent which does not adversely influence the reaction. A liquid base or acid can be also used as a solvent.

The reaction temperature is not critical and the reaction is usually carried out under cooling to warming.

Process 3-(i) The compound [VI] or a salt thereof can be prepared by reacting the compound [IV] or a salt thereof with the compound or a salt thereof.

Suitable salt of the compounds [IV] can be referred to the ones as exemplified for the compound Suitable salts of the compounds include an organic acid addition salt formate, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc.], an inorganic acid addition salt hydrochloride, hydrobromide, sulfate, phosphate, etc.] and the like.

The present reaction may be carried out in a solvent such as water, phosphate buffer, acetone, chloroform, P:\OPER\KbM\specis\12628860 specidoc acetonitrile, nitrobenzene, methylene chloride, ethylene Schloride, formamide, N,N-dimethylformamide, methanol, ethanol, diethyl ether, tetrahydrofuran, dimethyl sulfoxide, Sor any other organic solvent which does not adversely affect the reaction, preferably in ones having strong polarities.

Among the solvents, hydrophilic solvents may be used in a mixture with water. When the compound is liquid, it can also be used as a solvent.

00 C The reaction is preferably conducted in the presence S 10 of a base, for example, an inorganic base such as alkali Smetal hydroxide, alkali metal carbonate, alkali metal hydrogencarbonate, an organic base such as trialkylamine, and the like.

The reaction temperature is not critical, and the reaction is usually carried out at ambient temperature, under warming or under heating. The present reaction is preferably carried out in the presence of alkali metal halide sodium iodide, potassium iodide, etc.], alkali metal thiocyanate sodium thiocyanate, potassium thiocyanate, etc.], and the like.

Anion ZO may be one derived from a leaving group Y, and it may be converted to other anion by a conventional method.

Process 3-(ii) The compound or a salt thereof can be prepared by subjecting the compound [VI] or a salt thereof to elimination reaction of the carboxy protecting group.

Elimination reaction is carried out in similar manner to the reaction in the aforementioned Process 2, and therefore the reagents to be used and reaction conditions solvent, reaction temperature, etc.) can be referred to those of Process 2.

Processes A and B for the preparation of the starting P:\OPER\Kbm\specis\12628860 specidoc compounds are explained in detail in the following.

Process A-(i) The compound [IX] or a salt thereof can be prepared by ;reacting the compound [VII] or a salt thereof with the compound [VIII] or a salt thereof.

This reaction can be carried out in a similar manner to the reaction in the aforementioned Process and Stherefore the reagents to be used and reaction conditions 00 c solvent, reaction temperature, etc.) can be referred 1 0 to those of Process 3-(i) Process A-(ii) The compound [II] or a salt thereof can be prepared by subjecting the compound [IX] or a salt thereof to elimination reaction of the amino protecting groups in R 11 and R 13 and the carboxy protecting group in R 12 This reaction can be carried out in a similar manner to the reaction in the aforementioned Process 2, and therefore the reagents to be used and reaction conditions solvent, reaction temperature, etc.) can be referred to those of Process 2.

Process B The compound [IV] or a salt thereof can be prepared by reacting the compound or its reactive derivative at the amino group, or a salt thereof with the compound [XI] or its reactive derivative at the carboxy group, or a salt thereof.

This reaction can be carried out in a similar manner to the reaction in the aforementioned Process i, and therefore the reagents to be used and reaction conditions solvent, reaction temperature, etc.) can be referred to those of Process 1.

The compounds obtained by the above processes can be isolated and purified by a conventional method such as pulverization, recrystallization, column chromatography, P:\OPER\Kb\specis\12628860 specidoc Sreprecipitation, and the like.

O It is to be noted that the compound and other C compounds may include one or more stereoisomer(s) such as Soptical isomer(s) and geometrical isomer(s) due to asymmetric carbon atom(s) and double bond(s), and all of such isomers and mixtures thereof are included within the scope of this invention.

The compounds and pharmaceutically acceptable 00 Ssalts thereof include solvates enclosure compounds hydrate, etc.)].

SThe compound and pharmaceutically acceptable salts thereof are novel and exhibit high antimicrobial activity, inhibiting the growth of a wide variety of pathogenic microorganisms including Gram-positive and Gram-negative microorganisms and are useful as antimicrobial agents.

Now in order to show the utility of the compound the test data on MIC (minimal inhibitory concentration) of a representative compound of this invention are shown in the following.

Test method: In vitro antibacterial activity was determined by the two-fold agar-plate dilution method as described below.

One loopful of an overnight culture of each test strain in Trypticase-soy broth (106 viable cells per ml) was streaked on heart infusion agar (HI-agar) containing graded concentrations of representative test compound, and the minimal inhibitory concentration (MIC) was expressed in tg/ml after incubation at 37 0 C for 20 hours.

Test compound Compound 7-{(Z)-2-(2-amino-5-chloro-l,3-thiazol-4-yl)- 2-[(carboxymethoxy)imino]acetamido}-3-[3-amino-4-(3aminopropionamido)-2-methyl-l-pyrazolio]methyl-3-cephem-4carboxylic acid hydrogen sulfate (Example 4B) P:\OPER\Kbm\specis\12628860 specidoc Compound 7p-{(Z)-2-(2-amino-5-chloro-l,3-thiazol-4-yl)- 2-[(carboxymethoxy)imino]acetamido}-3-[3-amino-4-(3aminopropyl)-2-methyl-l-pyrazolio]methyl-3-cephem-4carboxylic acid hydrogen sulfate (Example Ceftazidime Test results: Table 2 Test strain Test compound MIC (Lg/ml) Pseudomonas aeruginosa 1 FP 1456 Ceftazidime 128 For therapeutic administration, the compound and pharmaceutically acceptable salts thereof of the present invention are used in the form of a conventional pharmaceutical preparation which contains said compound as an active ingredient, in admixture with pharmaceutically acceptable carriers such as an organic or inorganic solid or liquid excipient which is suitable for oral, parenteral or external administration. The pharmaceutical preparations may be in a solid form such as tablet, granule, powder, capsule, or in a liquid form such as solution, suspension, syrup, emulsion, lemonade and the like.

If needed, there may be included in the above preparations auxiliary substances, stabilizing agents, wetting agents and other commonly used additives such as lactose, citric acid, tartaric acid, stearic acid, magnesium stearate, terra alba, sucrose, corn starch, talc, gelatin, agar, pectin, peanut oil, olive oil, cacao butter, ethylene glycol, and the like.

While the dosage of the compound may vary from and also depend upon the age, conditions of the patient, a kind P:\OPER\Kbm\specis\12628860 specidoc of diseases, a kind of the compound to be applied, etc.

O In general amounts between 1 mg and 4,000 mg or even more C per day may be administered to a patient. An average single Sdose of about 50 mg, 100 mg, 250 mg, 500 mg, 1000 mg or 2000 5 mg of the compounds of the present invention may be used in treating diseases infected by pathogenic microorganisms.

The following Preparations and Examples are given for Sthe purpose of illustrating the present invention in more detail.

Preparation 1B To a solution of 5-amino-l-methylpyrazole (5 g) in ethanol (50 ml) was added isoamyl nitrite (6.92 ml), and then 20% hydrochloric acid (5 drops) was added at 4 0 C. The reaction mixture was refluxed for 2 hours and cooled to room temperature. To the reaction mixture was added diisopropyl ether (50 ml), and the mixture was stirred for 0.5 hour.

The resulting precipitate was collected by filtration and dried in vacuo to give 5-amino-l-methyl-4-nitrosopyrazole (3.53 g).

1 H-NMR(DMSO-d) 6 3.51 (3H, 8.07 (2H, brs), 8.51 (1H, s) APCI-MS: m/z=127(M+H) Preparation 2B To a solution of 5-amino-l-methyl-4-nitrosopyrazole (1 g) in water (40 ml) were added concentrated sulfuric acid (0.423 ml) and palladium on carbon (0.3 g) under a hydrogen atmosphere. The mixture was stirred overnight. The reaction mixture was filtered, and the filtrate was evaporated in vacuo. To the residue was added isopropyl alcohol, and the resulting precipitate was collected by filtration to give 4,5-diamino-l-methylpyrazole sulfate (1.71 g).

1 H-NMR(DMSO-d 6 6 3.54 (3H, 7.19 (1H, s) ESI-MS: m/z=113(M+H) P:\OPER\K1m\specis\12628860 speci.doc c Preparation 3B O To a suspension of sulfate (305 g) in tetrahydrofuran (3.05 L) was added tertbutyl {2-[(2,5-dioxo-1-pyrrolidinyl)oxy]-2oxoethyl}carbamate (415 g) under ice-cooling. To the mixture was added diisopropylethylamine (556 ml) dropwise at a temperature below 10 0 C. The mixture was stirred at room 00 temperature overnight. To the resulting solution were added brine and saturated aqueous sodium hydrogen carbonate n 10 solution, and the mixture was extracted with ethyl acetate The aqueous layer was extracted with tetrahydrofuran/ethyl acetate=1/l (3.0 L) twice. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl ether (1.0 L) and dried in vacuo to give tert-butyl {2-[(5-amino-l-methylpyrazol-4-yl)amino]- 2-oxoethyl}carbamate (307 g).

IR(KBr) 3440, 3349, 1670, 1631, 1525., 1276, 1163, 1074, 1014, 860, 791 cm 1 'H-NMR(DMSO-d6) 6 1.39 (9H, 3.44 (3H, 3.64 (2H, d, J 5.9 Hz), 4.91 (2H, brs), 6.97 (1H, t, J 5.9 Hz), 7.15 (1H, 9.09 (1H, brs) Preparation 4B To a solution of tert-butyl methylpyrazol-4-yl)amino]-2-oxoethyl}carbamate (307 g) in N,N-dimethylformamide (1.5 L) was added trityl chloride (334 To the mixture was added triethylamine (318 ml) dropwise. The mixture was stirred at room temperature for 1 hour. The reaction mixture was dissolved in ethyl acetate.

The solution was washed successively with water, 10% aqueous citric acid solution, water and brine. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with P:\OPER\KbM\SpeCiS\1262886O epecidoc acetonitrile (1.5 L) and dried in vacuo to give tert-butyl.

[1-methyl-5-(tritylamino)-1H-pyrazol-4-yllamino}-2oxoethyl)carbamate (468 g).

;Z IR(KBr) 3336, 3280, 1724, 1683, 1599, 1234, 939, 704 cm 1 1 H-NMR (DMSO-d 6 8 1.38 (9H, s) 2. 73 (3H, s) 3.38 (2H, d, J =5.8Hz), 5.58 (1H, 6.94 (1H, t, J 5.8 Hz), 7.11-7.35 in), 7.21 (1H, 8.31 (1H, s) 00 ESI-MS: m/z=512.3(M+H+) Preparation tert-Butyl [(5-amino-1-methylpyrazol-4-yl)amino] -3oxopropyl }carbamate The title compound was obtained from 4,5-diamino-1methylpyrazole sulfate and tert-butyl 5-dioxo-1pyrrolidinyl)oxy]-3-oxopropyl~carbamate in the same manner as in Preparation 3B.

1 H-NMR (DMSO-d 6 61. 38 (9H, s) 2. 35 (2H, t, J 7. 1 Hz), 3.18 (2H, q, J =7.1 Hz), 3.50 (3H, 4.90 (2H, 6.83 (1H, t, J 7.1 Hz), 7.14 (1H, 9.06 (1H, s) AP-MS: m/z=283 Preparation 6B tert-Butyl [1-methyl-5- (tritylamino)pyrazol-4y1] amino} -3-oxopropyl) carbamate The title compound was obtained from tert-butyl {3- II(5-amino-1-methylpyrazol-4-y1) amino] -3-oxopropyl~carbamate in the same manner as in Preparation 4B.

'H-NMR (DMSO-d 6 6 1. 39 (9H, s) 2. 08 (2H, t, J 7. 1 Hz) 2.71 (3H, 3.04 (2H, q, J 7.1 Hz), 5.57 (1H, 6.72 (1H, t, J 7.1 Hz), 7.1-7.4 (16H, in), 8.25 (1H, s) Preparation 7B To a suspension of 5-amino-1-methylpyrazole-4carbaldehyde (25 g) and triethylamine (22.2 g) in dichioromethane (500 ml) was added trityl chloride (61.3 g) at room temperature. The mixture was stirred at room P:\OPER\Kbm\specis\12628860 specidoc temperature for 70 hours. The reaction mixture was washed with successively 10% aqueous citric acid solution, brine c and 10% aqueous sodium hydrogen carbonate solution. The Sextract was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The residue was triturated with ethyl acetate to give (tritylamino)pyrazole-4-carbaldehyde (67.6 g) as a colorless Ssolid.

00

S

1 H-NMR(CDC13) 8 2.84 (3H, 7.26-7.34 (15H, 7.60 (1H, 8.95 (1H, brs), 9.58 (1H, s) SPreparation 8B To a suspension of sodium hydride (55% dispersion in mineral oil, 26.2 g) in tetrahydrofuran (1000 ml) was added diethyl (cyanomethyl)phosphonate (106.3 g) under icecooling. The mixture was stirred under ice-cooling for minutes. To the mixture was added tritylaminopyrazole-4-carbaldehyde (197 and the mixture was stirred for 2 hours. The reaction mixture was poured into ice-cold water. The mixture was extracted with ethyl acetate. The extract was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The residue was triturated with diethyl ether to give 3-[1-methyl-5- (tritylamino)pyrazol-4-yl]-2-propenonitrile (E,Z mixture, 126 g) as a solid.

E form 1H-NMR(CDC13) 5 3.14 (3H, 4.45 (1H, br), 5.09 (1H, d, J 16.5 Hz), 6.43 (1H, d, J 16.5 Hz), 7.17-7.32 (15H, m), 7.42 (1H, s) Z form 'H-NMR(CDC13) 8 3.09 (3H, 4.37 (1H, br), 4.58 (1H, d, J 11.9 Hz), 6.22 (1H, d, J 11.9 Hz), 7.17-7.32 (15H, m), 8.14 (1H, s) Preparation 9B P:\OPER\Kbm\speCis\12628860 specidoc To a mixture of lithium aluminum hydride (32.6 g) in tetrahydrofuran (1.3 L) was added (2E)-3-[l-methyl-5c (tritylamino)pyrazol-4-yl]-2-propenonitrile (101.6 g) under Sice-cooling. The mixture was refluxed for 4 hours. After cooling on an ice bath, sodium fluoride (100 g) and water (100 ml) were added to the reaction mixture. The insoluble materials were removed by filtration. The filtrate was Sconcentrated in vacuo to give 4-(3-aminopropyl)-l-methyl-5- 00 c (tritylamino)pyrazole (88.9 g).

S 10 H-NMR(DMSO-d 6 6 1.00-1.22 (2H, 1.70-1.90 (2H, 2.15- 2.35 (2H, 2,74 (3H, 5.70 (1H, 6.97 (1H, s), 7.10-7.38 (15H, m) ESI-MASS: m/z=397.4 (M+H) Preparation To a solution of 4-(3-aminopropyl)-l-methyl-5- (tritylamino)pyrazole (75 g) in tetrahydrofuran (700 ml) was added di-tert-butyl dicarbonate (49.5 The reaction mixture was stirred at room temperature for 3 hours. After evaporation of the solvent in vacuo, the residue was triturated with diisopropyl ether and dried in vacuo to give tert-butyl {3-[l-methyl-5-(tritylamino)pyrazol-4yl]propyl}carbamate (71.7 g).

1 H-NMR(DMSO-d 6 6 1.10-1.25 (2H, 1.39 (9H, 1.65-1.85 (2H, 2.60-2.80 (2H, 2.75 (3H, 5.66 (1H, s), 6.50-6.70 (1H, 6.99 (1H, 7.05-7.35 (15H, m) Preparation 11B To a suspension of l,l'-carbonyldiimidazole (9.73 g) in dehydrated chloroform (72 ml) was added tert-butyl N-(2aminoethyl)carbamate (9.61 g) under ice-cooling, and the mixture was stirred at room temperature for 1 hour. To the reaction mixture were added N-ethyldiisopropylamine (14.22 g) and 4,5-diamino-l-methylpyrazole sulfate (10.51 and the mixture was stirred at 50 0 C for 15 hours. The insoluble P:\OPER\Kbm\specis\12628860 specidoc materials were removed by filtration. To the filtrate were Sadded chloroform (200 ml) and 5% aqueous sodium hydrogen carbonate solution (100 ml). The organic layer was Sseparated, and the aqueous layer was extracted with a mixed solvent of chloroform and methanol The organic layers were combined, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue O was triturated with ethyl acetate and dried in vacuo to give 00 C tert-butyl [2-({[(5-amino-l-methylpyrazol-4yl)amino]carbonyl}amino)ethyl]carbamate (14.0 g) as a solid.

1 H-NMR(DMSO-d 6 6 1.38 (9H, 2.96-2.98 (2H, 3.03-3.07 (2H, 3.50 (3H, 4.81 (2H, br), 5.92 (1H, br), 6.80 (1H, br), 6.96 (1H, 7.18 (1H, br) Preparation 12B To a solution of tert-butyl methylpyrazol-4-yl)amino]carbonyl}amino)ethyl]carbamate (597 mg) and triethylamine (243 mg) in methylene chloride (10 ml) was added trityl chloride (669 mg), and the mixture was stirred at room temperature for 19 hours. The reaction mixture was washed successively with 10% aqueous citric acid solution, brine and saturated aqueous sodium hydrogen carbonate solution. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with ethyl acetate to give tert-butyl {2-[({[1-methyl-5-(tritylamino)pyrazol-4yl]amino}carbonyl)amino]ethyl}carbamate (640 mg) as a solid.

1H-NMR(DMSO-d 6 5 1.38 (9H, 2.70 (3H, 2.94-2.96 (2H, 2.99-3.01 (2H, 5.68 (1H, brs), 5.96 (1H, br), 6.78 (1H, br), 6.85 (1H, br), 7.00 (1H, 7.13-7.15 (6H, m), 7.24-7.28 (9H, m) Preparation 13B To a solution of methyl (Z)-2-hydroxyimino-2-[5methyl-2-(tritylamino)-1,3-thiazol-4-yl]acetate (11.38 g) in P: \OPER\Kbm\specis\1262880 speci.doc tetrahydrofuran (170 ml) was added sodium hydride Sdispersion in mineral oil, 1.20 g) at room temperature. The mixture was stirred at the same temperature for 20 minutes.

STo the reaction mixture was added tert-butyl bromoacetate (3.68 ml) and the mixture was stirred at room temperature for 3 hours. To the reaction mixture were added water and ethyl acetate. The aqueous layer was separated and the Sorganic layer was washed with brine. The organic layer was 00 dried over anhydrous magnesium sulfate, filtered, and S 10 concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with ethyl acetatehexane to give methyl (Z)-2-[(tertbutoxycarbonylmethoxy)imino]-2-[5-methyl-2-(tritylamino)- 1,3-thiazol-4-yl]acetate (6.08 g) as a solid.

1 H-NMR (DMSO-d 6 5 1.39 (9H, 2.22 (3H, 3.32 (3H, brs), 4.51 (2H, 7.10-7.35 (15H, m) 8.54 (1H, s) Preparation 14B To a solution of methyl (Z)-2-[(tertbutoxycarbonylmethoxy)imino]-2-[5-methyl-2-(tritylamino)- 1,3-thiazol-4-yl]acetate (1.72 g) in pyridine (12 ml) was added lithium iodide (4.02 g) at room temperature. The mixture was stirred at 50-80 0 C for 4 hours. To the reaction mixture were added ethyl acetate and 5% aqueous citric acid solution. The aqueous layer was separated and the organic layer was washed successively with water, 5% aqueous citric acid solution and brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was triturated with ethyl acetate and dried in vacuo to give (Z)-2-[(tertbutoxycarbonylmethoxy)imino]-2-[5-methyl-2-(tritylamino)- 1,3-thiazol-4-yl]acetic acid (936 mg) as a solid 1 H-NMR (DMSO-d 6 5 1.40 (9H, 2.18 (3H, 4.51 (2H, s), 7.10-7.35 (15H, m) 8.48 (1H, s) P:\OPER\Kbm\speciS\12628860 speci.doc SPreparation O To a solution of (Z)-2-[(tert- C 1 butoxycarbonylmethoxy)imino]-2-[5-methyl-2-(tritylamino)- S1,3-thiazol-4-yl]acetic acid (1.12 g) in N,N- 5 dimethylacetamide (8 ml) was added potassium carbonate (276 mg). To the mixture was added dropwise methanesulfonyl C, chloride (310 jil) under ice-cooling and the mixture was OO stirred at the same temperature for 40 minutes. To the (Ni Sreaction mixture were added 4-methoxybenzyl 70-amino-3- V) 10 chloromethyl-3-cephem-4-carboxylate hydrochloride (885 mg) Sand potassium carbonate (276 mg) and the mixture was stirred at the same temperature for 90 minutes. To the reaction mixture were added water and ethyl acetate. The aqueous layer was separated and the organic layer was washed with brine. The organic layer was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (120 ml), and the resulting precipitate was collected by filtration and dried in vacuo to give crude 4-methoxybenzyl 7p-{(Z)-2-[(tertbutoxycarbonylmethoxy)imino]-2-[5-methyl-2-(tritylamino)- 1,3-thiazol-4-yl]acetamido}-3-chloromethyl-3-cephem-4carboxylate (1.17 This product was used in the next step without further purification.

Example 1B To a solution of 4-methoxybenzyl 7p-{(Z)-2-[(tertbutoxycarbonylmethoxy)imino]-2-[5-methyl-2-(tritylamino)- 1,3-thiazol-4-yl]acetamido}-3-chloromethyl-3-cephem-4carboxylate (1.15 g) in N,N-dimethylformamide (3.0 ml) was added N-(trimethylsilyl)acetamide (840 mg) at room temperature, and the mixture was stirred at the same temperature for 30 minutes. To the reaction mixture was added potassium iodide (297 mg), and the mixture was stirred P:\OPER\Kbm\speCiS\12628860 specidoc at the same temperature for 30 minutes. To the reaction O mixture was added tert-butyl (3-{[1-methyl-5- (tritylamino)pyrazol-4-yl]amino}-3-oxopropyl)carbamate (807 Smg) at the same temperature. The whole mixture was stirred at the same temperature for 4 hours. To the resulting reaction mixture was added ethyl acetate and the solution was washed with brine, dried over sodium sulfate and Sfiltered. The filtrate was concentrated to about 10 ml in 00 vacuo. The concentrate was poured into diisopropyl ether (100 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene chloride (3.0 ml) were added anisole (1.0 ml) and trifluoroacetic acid (2.0 ml) at room temperature. The resulting solution was stirred at the same temperature for 3 hours and poured into diisopropyl ether (100 ml). The resulting precipitate was collected by filtration and dried in vacuo to give a crude product, which was purified by preparative high-performance liquid chromatography (HPLC) utilizing ODS column. The eluate containing a desired product was concentrated to about 30 ml in vacuo. The concentrate was further purified by preparative HPLC utilizing ODS column eluting with 8% aqueous acetonitrile. The eluate was concentrated to about ml in vacuo and lyophilized to give 7p-{(Z)-2-(2-amino-5methyl-l,3-thiazol-4-yl)-2- [(carboxymethoxy)imino]acetamido}-3-[3-amino-4-(3aminopropionamido)-2-methyl-l-pyrazolio]methyl-3-cephem-4carboxylate (17.3 mg) as an amorphous solid.

1 H-NMR (D 2 0) 8 2.34 (3H, 2.88 (2H, t, J 6.5 Hz), 3.18 (1H, d, J 17.5 Hz), 3.32 (2H, t, J 6.5 Hz), 3.43 (1H, d, J 17.5 Hz), 3.72 (3H, 4.55 (2H, 5.00 (1H, d, J 15.5 Hz), 5.17 (1H, d, J 15.5 Hz), 5.22 (1H, d, J 5 Hz), 5.85 (1H, d, J 5 Hz), 8.00 (1H, s) P \OPER\Kbm\SpeCis\12628860 specidoc SPreparation 16B STo a suspension of (Z)-2-[(tertbutoxycarbonylmethoxy)imino]-2-(2-formamido-1,3-thiazol-4- Syl)acetic acid (990 mg) in methanol (10 ml) was added Nbromosuccinimide (534 mg) at room temperature. The mixture was stirred at the same temperature for 5 minutes. To the reaction mixture were added saturated aqueous sodium Shydrogen carbonate solution and chloroform. The aqueous 00 layer was separated and the organic layer was dried over anhydrous magnesium sulfate, filtered, and concentrated in vacuo. The residue was triturated with diethyl ether and

C

dried in vacuo to give (Z)-2-(5-bromo-2-formamido-l,3thiazol-4-yl)-2-[(tert-butoxycarbonylmethoxy)imino]acetic acid (770 mg) as a solid.

'H-NMR (DMSO-d 6 8 1.43 (9H, 4.48 (2H, 8.51 (1H, s) Preparation 17B To a solution of (Z)-2-(5-bromo-2-formamido-l,3thiazol-4-yl)-2-[(tert-butoxycarbonylmethoxy)imino]acetic acid (816 mg) in N,N-dimethylacetamide (8 ml) was added potassium carbonate (276 mg). To the mixture was added dropwise methanesulfonyl chloride (310 pL) under ice-cooling and the mixture was stirred at the same temperature for minutes. To the reaction mixture were added 4-methoxybenzyl 70-amino-3-chloromethyl-3-cephem-4-carboxylate hydrochloride (810 mg) and potassium carbonate (276 mg), and the mixture was stirred at the same temperature for 90 minutes. To the reaction mixture were added water and ethyl acetate. The aqueous layer was separated and the organic layer was washed with brine. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (120 ml), and the resulting precipitate was collected by filtration and dried in vacuo P:\OPER\Kbm\speciS\12626860 specidoc qI to give crude 4-methoxybenzyl 7-{(Z)-2-(5-bromo-2formamido-1,3-thiazol-4-yl)-2-[(tertbutoxycarbonylmethoxy)imino]acetamido}-3-chloromethyl-3cephem-4-carboxylate (870 mg). This product was used in the 5 next step without further purification.

(N

Example 2B Cy- To a solution of 4-methoxybenzyl 73-{(Z)-2-(5-bromo-2- 00 formamido-1,3-thiazol-4-yl)-2-[(tertcN Sbutoxycarbonylmethoxy)imino]acetamido}-3-chloromethyl-3c-i V) 10 cephem-4-carboxylate (2.96 g) in N,N-dimethylformamide (8.9 Sml) was added N-(trimethylsilyl)acetamide (2.56 g) at room temperature, and the mixture was stirred at the same temperature for 30 minutes. To the reaction mixture was added potassium iodide (906 mg), and the mixture was stirred at the same temperature for 30 minutes. To the reaction mixture was added tert-butyl (tritylamino)pyrazol-4-yl]amino}-3-oxopropyl)carbamate at the same temperature. The whole mixture was stirred at the same temperature for 4 hours. To the resulting reaction mixture was added ethyl acetate and the mixture was washed with 10% aqueous sodium trifluoroacetate solution and brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated to about 10 ml in vacuo. The concentrate was poured into diisopropyl ether (250 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid (3.2 g) in methylene chloride (9.0 ml) were added anisole ml) and trifluoroacetic acid (6.0 ml) at room temperature.

The resulting solution was stirred at the same temperature for 14 hours and poured into diisopropyl ether (250 ml).

The resulting precipitate was collected by filtration and dried in vacuo to give a crude product (2.4 which was purified by preparative HPLC utilizing ODS column. The P:\OPER\Kbm\speciS\1262886O specidoc eluate containing a desired product was concentrated to Sabout 30 ml in vacuo. The concentrate was further purified C by preparative HPLC utilizing ODS column eluting with Saqueous acetonitrile. The eluate was concentrated to about 5 10 ml in vacuo and lyophilized to give 7p-{(Z)-2-(5-bromo-2-

(N

formamido-1,3-thiazol-4-yl)-2- I [(carboxymethoxy)imino]acetamido}-3-[3-amino-4-(3- 00 aminopropionamido)-2-methyl-l-pyrazolio]methyl-3-cephem-4- Scarboxylate (147 mg) as an amorphous solid.

in 10 H-NMR (D 2 0) 8 2.87 (2H, t, J 6.5 Hz), 3.17 (1H, d, J S17.5 Hz), 3.31 (2H, t, J 6.5 Hz), 3.43 (1H, d, J 17.5

(N

Hz), 3.72 (3H, 4.62 (2H, 5.00 (1H, d, J 15.5 Hz), 5.16 (1H, d, J 15.5 Hz), 5.23 (1H, d, J 5 Hz), 5.87 (1H, d, J 5 Hz), 8.01 (1H, 8.53 (1H, s) Example 3B A solution of 7p-{(Z)-2-(5-bromo-2-formamido-1,3thiazol-4-yl)-2-[(carboxymethoxy)imino]acetamido}-3-[3amino-4-(3-aminopropionamido)-2-methyl-l-pyrazolio]methyl-3cephem-4-carboxylate (130 mg) in 1M hydrochloric acid (13 ml) was stirred at room temperature for 6 hours. The reaction mixture was adjusted to about pH 7 by addition of aqueous dipotassium hydrogenphosphate solution. The resulting solution was purified by preparative HPLC utilizing ODS column eluting with 6% aqueous acetonitrile.

The eluate was concentrated to about 10 ml in vacuo and 0.05M aqueous sulfuric acid solution (2.18 ml) was added.

The resulting solution was lyophilized to give amino-5-bromo-l,3-thiazol-4-yl)-2- [(carboxymethoxy)imino]acetamido}-3-[3-amino-4-(3aminopropionamido)-2-methyl-l-pyrazolio]methyl-3-cephem-4carboxylic acid hydrogen sulfate (114 mg) as an amorphous solid.

1H-NMR (D 2 0) 8 2.88 (2H, t, J 6.5 Hz), 3.21 (1H, d, J P:\OPER\Kbm\speci\12628860 specidoc 17.5 Hz), 3.32 (2H, t, J 6.5 Hz), 3.43 (1H, d, J 17.5 O Hz), 3.72 (3H, 5.02 (1H, d, J 15.5 Hz), 5.23 (1H, d, J 15.5 Hz), 5.23 (1H, d, J 5 Hz), 5.88 (1H, d, J 5 Hz), S8.02 (1H, s) Preparation 18B To a solution of (Z)-2-(2-amino-5-chloro-l,3-thiazol- 4 -yl)-2-[(tert-butoxycarbonylmethoxy)imino]acetic acid (1.55 g) in N,N-dimethylacetamide (8 ml) were added methansulfonyl 00 chloride (0.716 ml) and potassium carbonate (639 mg) at 0 C V) 10 and the mixture was stirred at the same temperature for 1 Shour. A solution of 4-methoxybenzyl 7p-amino-3chloromethyl-3-cephem-4-carboxylate hydrochloride (1.50 g) and N-(trimethylsilyl)acetamide (4.86 g) in N,Ndimethylacetamide (9 ml), which had been stirred at 35 0 C for 1 hour, was added to the reaction mixture obtained above at 0 0 C. The whole mixture was stirred at 0 C for 1 hour.

Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The extract was washed with water and brine, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure to give 4methoxybenzyl 7p-{(Z)-2-(2-amino-5-chloro-l,3-thiazol-4-yl)- 2-[(tert-butoxycarbonylmethoxy)imino]acetamido}-3chloromethyl-3-cephem-4-carboxylate (1.50 g) as a solid.

1H-NMR (DMSO-d 6 6 1.42 (9H, 3.51 and 3.71 (2H, ABq, J 18.1 Hz), 3.75 (3H, 4.44 and 4.55 (2H, ABq, J 11.4 Hz), 4.56 (2H, 5.14-5.28 (3H, 5.86 (1H, dd, J 8.7, 4.9 Hz), 6.93 (2H, d, J 8.6 Hz), 7.36 (2H, d, J 8.6 Hz), 7.42 (2H, brs), 9.53 (1H, d, J 8.7 Hz) MS(ESI) 704.1 (M+Na+) Example 4B To a solution of 4-methoxybenzyl 7-{(Z)-2-(2-amino-5chloro-l, 3-thiazol-4-yl)-2-[(tertbutoxycarbonylmethoxy)imino]acetamido}-3-chloromethyl-3- 77 P:\OPER\Kbn\speci\12628860 speci.doc cephem-4-carboxylate (750 mg) in N,N-dimethylformamide (2.25 Sml) was added 1,3-bis(trimethylsilyl)urea (1.12 and the C mixture was stirred at room temperature for 30 minutes. To Sthe reaction mixture was added potassium iodide (199 mg), and the mixture was further stirred for 30 minutes. To the resulting mixture was added tert-butyl (tritylamino)pyrazol-4-yl]amino}-3-oxopropyl)carbamate (861 mg) and the whole mixture was stirred at 35-40 0 C overnight.

To the reaction mixture was added ethyl acetate and the n 10 mixture was washed successively with water, aqueous Ssodium thiosulfate solution, and 10% aqueous trifluoroacetic acid solution, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether, and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene chloride (3.3 ml) were added anisole (1.1 ml) and trifluoroacetic acid (3.3 ml).

The resulting solution was stirred at room temperature for 4 hours and poured into diisopropyl ether. The resulting precipitate was collected by filtration and dried in vacuo to give a crude product, which was purified by preparative HPLC utilizing ODS column. The eluate containing a desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to about pH 3 by addition of concentrated hydrochloric acid and chromatographed on Diaion (registered trademark) (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 10 ml in vacuo and 0.05M aqueous sulfuric acid solution (2.18 ml) was added. The resulting solution was lyophilized to give 73- {(Z)-2-(2-amino-5-chloro-l,3-thiazol-4-yl)-2- [(carboxymethoxy)imino]acetamido}-3-[3-amino-4-(3- P:\OPER\Kbm\specis\12628860 specidoc aminopropionamido) -2-methyl-l-pyrazolio] methyl-3-cephem-4carboxylic acid hydrogen sulfate (61 mg) as an amorphous solid.

IR(KBr) 1776, 1670, 1541, 1109 cm- 1 1 H-NMR (D 2 0) 8 2.89 (2H, t, J 6.5 Hz), 3.20 and 3.43 (2H, ABq, J 17.9 Hz), 3.33 (2H, t, J 6.5 Hz), 3.71 (3H, s), 4.78 (2H, 5.01 and 5.24 (2H, ABq, J 15.8 Hz), 5.22 00 (LH, d, J 4.9 Hz), 5.87 (LH, d, J 4.9 Hz), 8.03 (1H, s).

(N

MS(ESI) 657.15 Example 7p-(()-2-(2-Amino-5-chloro-1,3-thiazol-4-yl)-2- [(carboxymethoxy)imino]acetamido}-3-[3-amino-4-(3aminopropyl)-2-methyl-l-pyrazolio]methyl-3-cephem-4carboxylic acid hydrogen sulfate The title compound was obtained as an amorphous solid in the same manner as in Example 4B using tert-butyl methyl-5-(tritylamino)pyrazol-4-yl]propyl~carbamate.

IR(KBr) 1774, 1647, 1113 cm 1 1H-NMR (D 2 0) 6 1.84-1.99 (2H, 2.48 (2H, t-like), 2.52 (2H, t-like), 3.16 and 3.36 (2H, ABq, J 17.7 Hz), 3.67 (3H, 4.78 (2H, 4.93 and 5.19 (2H, ABq, J 16.0 Hz), 5.21 (1H, d, J 4.9 Hz), 5.86 (LH, d, J 4.9 Hz), 7.77 (1H, s) MS(ESI) 628.24 Example 6B 71-((Z)-2-(2-Amino-5-chloro-l,3-thiazol-4-yl)-2- [(carboxymethoxy)imino]acetamido}-3-[3-amino-4({[(2aminoethyl)amino]carbonyl~amino)-2-methyl-ipyrazolio]methyl-3-cephem-4-carboxylic acid hydrogen sulfate The title compound was obtained as an amorphous solid in the same manner as in Example 4B using tert-butyl {2- [(([l-methyl-5-(tritylamino)pyrazol-4yl]amino~carbonyl)amino]ethyl~carbamate, except that 1- P:\OPER\Kbm\specis\12628860 specidoc methyl-2-pyrrolidone was used as a solvent instead of N,Ndimethylformamide.

IR(KBr) 1776, 1647, 1543, 1111 cm- 1 ;Z I~H-NMR (D 2 0) 5 3. 13 (2H, t, J 5. 6 Hz) 3. 22 and 3.48 (2H, ABq, J 17.8 Hz), 3.48 (2H, t, J 5.6 Hz), 3.71 (3H, s), 4.77 (2H, 5.00 and 5.22 (2H, ABq, J 15.7 Hz), 5.23 (1H, d, J 4.8 Hz), 5.88 (1H, d, J 4.8 Hz), 7.92 (1H, s) 00 MS(ESI) 672.28 00i Preparation 19B To a solution of tert-butyl (2S)-2-[(l.3-dioxo-l,3dihydro-2H-isoindol-2-yl)oxylpropanoate (1.5 g) in ethanol ml) was added hydrazine monohydrate (0.25 ml) at 0 0

C

and the mixture was stirred at room temperature for 1 hour.

The resulting precipitate was filtered off and the filtrate was evaporated in vacuo. The residue was dissolved in ethanol (12 ml) and tetrahydrofuran (12 ml). To the solution was added 2- (5-chloro-2-formamido-l,3--thiazol-4yl)-2-oxoacetic acid (1.21 and the mixture was stirred at room temperature for 2 hours. The volatiles were evaporated under reduced pressure and the residue was purified by silica gel column chromatography to give (2Z)-2- [(lS)-2-tert-butoxy-l-methyl-2-oxoethoxylimino)-2-(5chloro-2-formamido-1,3-thiazol-4-yl)acetic acid (1.31 g) as a solid.

Only the data of the major rotamer is described.

1 H-NMR (DMSO-d 6 8 1.36 (3H, d, J 7. 0 Hz) 1. 41 (9H, s), 4.65 (1H, q, J 7.0 Hz), 7.45 (1H, 8.55 (1H, 12.89 (1H, brs) MS(ESI negative) 376.1 Preparation 47Methoxybenzyl -2-tert-butoxy-lmethyl-2-oxoethoxy] imino}-2- (5-chloro-2-formamido-l,3thiazol-4-yl) acetamido] -3-chloromethyl-3-cephem-4- P:\OPER\Kbm\speciS\12628860 apeci.doc carboxylate O The title compound was obtained as a solid in the same C manner as in Preparation 18B using (2Z)-2-{[(1S)-2-tertbutoxy-l-methyl-2-oxoethoxy]imino}-2-(5-chloro-2-formamido- 1,3-thiazol-4-yl)acetic acid.

1 H-NMR (DMSO-d 6 8 1.41 (3H, t-like), 1.42 (9H, 3.47-3.80 (2H, 3.76 (3H, 4.46 and 4.55 (2H, ABq, J 11.4 Hz), O0 4.49-4.66 (1H, 5.14-5.29 (3H, 5.79-5.96 (1H, m), S6.94 (2H, d, J 8.7 Hz), 7.33-7.41 (1H, 7.36 (2H, d, J 8.7 Hz), 8.53 (1H, s) SExample 7B To a solution of 4-methoxybenzyl tert-butoxy-l-methyl-2-oxoethoxy]imino}-2-(5-chloro-2formamido-1,3-thiazol-4-yl)acetamido]-3-chloromethyl-3cephem-4-carboxylate (934 mg) in N,N-dimethylformamide (2.8 ml) was added 1,3-bis(trimethylsilyl)urea (1.31 and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added potassium iodide (234 mg), and the mixture was further stirred for 30 minutes. To the resulting mixture was added tert-butyl (tritylamino)pyrazol-4-yl]amino}-3-oxopropyl)carbamate (1.01 g) and the whole mixture was stirred at 35-40 0 C overnight.

To the reaction mixture was added ethyl acetate and the mixture was washed successively with water, aqueous sodium thiosulfate solution, and 10% aqueous trifluoroacetic acid solution, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether, and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene chloride (3.3 ml) were added anisole (1.1 ml) and trifluoroacetic acid (3.3 ml).

The resulting solution was stirred at room temperature P:\OPER\Kbm\speci\12628860 specidoc for 4 hours and poured into diisopropyl ether. The O resulting precipitate was collected by filtration and dried Cl in vacuo to give a crude product. The crude product was Sdissolved in methanol (5 ml). To the solution was added concentrated hydrochloric acid (0.1 ml) and the mixture was stirred at room temperature for 3 hours. After neutralization with sodium hydrogen carbonate and water, Smethanol was evaporated off and the residual aqueous 00 C solution was purified by preparative HPLC utilizing ODS S 10 column. The eluate containing a desired product was Sconcentrated to about 30 ml in vacuo. The concentrate was c adjusted to about pH 3 by addition of concentrated hydrochloric acid and chromatographed on Diaion (registered trademark) HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 10 ml in vacuo and lyophilized to give amino-5-chloro-l,3-thiazol-4-yl)-2-{[(lS)-1carboxyethoxy]imino}acetamido]-3-[3-amino-4-(3aminopropionamido)-2-methyl-1-pyrazolio]methyl-3-cephem-4carboxylic acid hydrogen sulfate (8 mg) as an amorphous solid.

1 H-NMR (D 2 0) 6 1.49 (3H, d, J 7.0 Hz), 2.88 (2H, t, J 6.4 Hz), 3.19 and 3.43 (2H, ABq, J 17.7 Hz), 3.32 (2H, t, J 6.4 Hz), 3.73 (3H, 4.85-5.00 (1H, 4.98 and 5.22 (2H, ABq, J 15.7 Hz), 5.23 (1H, d, J 4.8 Hz), 5.86 (1H, d, J 4.8 Hz), 8.02 (1H, s) Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

P:\OPER\Kbm\speciu\12628860 specidoc The reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that that prior art forms part of the Scommon general knowledge in Australia.

00 oO Oq

Claims (26)

1. A compound of the formula H3C CH3 O R RHoR NC wherein R 1 is carboxy or protected carboxy, R 2 is amino or protected amino, and R 3 is R 4 -Y R R 6 R 7 O N\ r N 8 or N wherein R 4 is hydrogen, amino or protected amino, R 5 is -(X)n-A-R 9 or a saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms, wherein X is -NHCO- or -NR' 1 (wherein R 10 is hydrogen or lower alkyl), A is lower alkylene, R 9 is amino or protected amino, and n is 0 or 1, R 6 and R 8 are each hydrogen, amino, protected amino or -(X)n-A-R 9 wherein X, A, R 9 and n are each as defined above, and R 7 is hydrogen, amino(lower)alkyl or protected amino(lower)alkyl, or a pharmaceutically acceptable salt thereof.

P:\OPER\Kbm\specis\12628860 specidoc S2. The compound of claim 1 wherein C( R 3 is R4 4 R wherein R 4 and R 5 are each as defined in claim 1, 00 0C or a pharmaceutically acceptable salt thereof.

3. The compound of claim 2 wherein 4 Cq R is hydrogen or amino, and R 5 is -(X)n-A-R 9 or piperazinyl, wherein X is -NHCO- or -NR 10 (wherein R 10 is hydrogen or lower alkyl), A is lower alkylene, R 9 is amino, and n is 0 or 1, or a pharmaceutically acceptable salt thereof.

4. A process for preparing a compound of the formula H 3 C CH 3 O R 1 N II S T C-CONH-- S 2A N -N CH 2 -R 3 [I] R s o COO wherein R 1 is carboxy or protected carboxy, R 2 is amino or protected amino, and R 3 is P:\OPER\Kbm\specis\12628860 specidoc in R 6 R 6 R 4 6/= N -N -NN -NN SR R R7 R8 or N R R R 8 R 7 wherein CA R 4 is hydrogen, amino or protected amino, 00 R 5 is -(X)n-A-R 9 or a saturated 3- to 8-membered heterocyclic S 5 group containing 1 to 4 nitrogen atoms, wherein X is -NHCO- or -NR 10 (wherein R 10 is Ohydrogen or lower alkyl), A is lower alkylene, R 9 is amino or protected amino, and n is 0 or 1, R 6 and R 8 are each hydrogen, amino, protected amino or -(X)n-A-R 9 wherein X, A, R 9 and n are each as defined above, and R 7 is hydrogen, amino(lower)alkyl or protected amino(lower)alkyl, or a salt thereof, which comprises reacting a compound of the formula [IV]: H 3 C CH 3 0 R 1 N II S [IV] N-T\-C-CONH-O- CH 2 [I 2X ,N N CH2-Y R S 0 1 1 wherein R 1 and R 2 are each as defined above, R 11 is carboxy or protected carboxy, and Y is a leaving group, or a salt thereof with a compound of the formula R 3 c [V] wherein R 3 c is a compound of the formula selected from P: \OPER\Kbm\speciS\12628860 speci.doc SR 6 R 6 R R N N N N N N NP-- R

5 N-R7 and N R 8 R 7 R R wherein R 4 R 5 R 6 R 7 and R 8 are each as defined above, C- optionally followed by deprotection, to give a compound of 00 the formula H 3 C CH VS O R l SOR (C N II S N-T C-CONH- S 2 .N 2A-N CH2-R 3 [I] R s e COO wherein R 1 R 2 and R 3 are each as defined above, or a salt thereof. A pharmaceutical composition comprising a compound of claim 1 or a pharmaceutically acceptable salt thereof in admixture with a pharmaceutically acceptable carrier.

6. A compound of claim 1 or a pharmaceutically acceptable salt thereof for use as a medicament.

7. A compound of claim 1 or a pharmaceutically acceptable salt thereof for use as an antimicrobial agent.

8. Use of a compound of claim 1 or a pharmaceutically acceptable salt thereof for manufacture of a medicament for treating infectious diseases.

9. A method for the treatment of infectious diseases which comprising administering a compound of claim 1 or a 87 P:\OPER\Kbm\speciS\12628860 speci.doc pharmaceutically acceptable salt thereof to human or animals.

A compound of the formula R R2 O R N 8 I IIN R RM S TT-C-CONHO CH2 4 S% R5 o N CH 2 N[I] R S R O cooN N COO 1 17 R R wherein R 1 and R 2 are each hydrogen, lower alkyl, hydroxy(lower)alkyl, protected hydroxy(lower)alkyl or halo(lower)alkyl, or R 1 and R 2 are bonded together and form lower alkylene; R 3 is carboxy or protected carboxy; R 4 is amino or protected amino; R 5 is hydrogen, halogen or lower alkyl; R 6 is lower alkyl which may have suitable substituent(s), R 7 is hydrogen or amino protecting group, or R 6 and R 7 are bonded together and form lower alkylene; R 8 is amino or protected amino; X is -NHCO-, -NHCONH- or -N(R 9 (wherein R 9 is hydrogen, lower alkyl or amino protecting group); A is lower alkylene; and n is 0 or 1, or a pharmaceutically acceptable salt thereof.

11. The compound of claim 10 wherein R 1 and R 2 are each hydrogen, lower alkyl, hydroxy(lower)alkyl or halo(lower)alkyl, or R 1 and R 2 are bonded together and form lower alkylene, or a pharmaceutically acceptable salt P:\OPER\Kbm\speciS\12628860 specidoc thereof. c

12. The compound of claim 10 wherein R 6 is lower alkyl, Sand R 7 is hydrogen, or a pharmaceutically acceptable salt thereof.

13. The compound of claim 10 wherein X is -NHCO- or NHCONH-, or a pharmaceutically acceptable salt thereof. 00

14. The compound of claim 10 wherein R 3 is carboxy, R 4 is amino, and R 8 is amino, or a pharmaceutically acceptable salt thereof.

A process for preparing a compound of the formula 1 2 R R 0 R N 8 N C- CONH- CH H [2 4EN- CH-N. R S 0 N N coo 16 17 R R wherein R 1 and R 2 are each hydrogen, lower alkyl, hydroxy(lower)alkyl, protected hydroxy(lower)alkyl or halo(lower)alkyl, or R 1 and R 2 are bonded together and form lower alkylene; R 3 is carboxy or protected carboxy; R 4 is amino or protected amino; R 5 is hydrogen, halogen or lower alkyl; R 6 is lower alkyl which may have suitable substituent(s), R 7 is hydrogen or amino protecting group, or R 6 and R 7 are bonded together and form lower alkylene; R 8 is amino or protected amino; P:\OPER\Kbm\specis\12628860 specidoc X is -NHCO-, -NHCONH- or -N(R 9 (wherein R 9 is hydrogen, O lower alkyl or amino protecting group); A is lower alkylene; and Sn is 0 or 1, or a salt thereof, which comprises reacting a compound of the formula [IV]: 1 2 CM R R O 3 00 O R NI S [IV] SN -C-CONH C- R R 5 N CH 2 R 1 0 wherein R 1 R 2 R 3 R 4 and R 5 are each as defined above, R 10 is protected carboxy, and Y is a leaving group, or a salt thereof with a compound of the formula -A-R 8 N H [V] N N I6 17 R R wherein R 6 R 7 R 8 X, A and n are each as defined above, or a salt thereof to give a compound of the formula [VI]: 1 2 R R 0 R N 8 II S. N CCONH R N R C-CONH N [VI R 1 0 6 17 R R wherein R R R R R, R R 7 R R 1 0 X, A and n are each as defined above, or a salt thereof, and subjecting the compound of the formula [VI] or a salt thereof to elimination reaction of the carboxy protecting P:\OPER\Kbm\specis\12628860 specidoc group, to give a compound of the formula 1 2 3 0 R N II N C-CONI (X)-A-R 8 4A/s 5 X 2 ,H 1 R S R N CH2 H [I] R S R 0Coo 2 N N coO 6 7 R R wherein R 1 R 2 R 3 R 4 R s R 6 R 7 R 8 X, A and n are each as defined above, or a salt thereof, or subjecting a compound of the formula [Ia]: 1 2 R R 2 0 R 3 N N C-CON] RXS R 5 Ba [Ia] N N 16 17 R R wherein R 1 R 2 R 3 R 4 R 5 R 6 R X, A and n are each as defined above, and R8a is protected amino, or a salt thereof to elimination reaction of the amino protecting group to give a compound of the formula [Ib]: 1 2 OR 3 O R i S (X)-A--NH 2 [Ib] N C-CONH 0 NT s R 5 o N c H 2 N% R S R 2 N COO I 7 R R wherein R 1 R 2 R 3 R 4 R 5 R 6 R X, A and n are each as defined above, or a salt thereof.

16. A pharmaceutical composition comprising a compound of P:\OPER\Kbm\specis\12628860 specidoc claim 10 or a pharmaceutically acceptable salt thereof in O admixture with a pharmaceutically acceptable carrier. F1

17. A compound of claim 10 or a pharmaceutically acceptable salt thereof for use as a medicament.

18. A compound of claim 10 or a pharmaceutically Sacceptable salt thereof for use as an antimicrobial agent. 00 S 10

19. Use of a compound of claim 10 or a pharmaceutically Sacceptable salt thereof for manufacture of a medicament for treating infectious diseases.

A method for the treatment of infectious diseases which comprising administering a compound of claim 10 or a pharmaceutically acceptable salt thereof to human or animals.

21. A compound according to claim 1 or claim substantially as hereinbefore described and/or exemplified.

22. A process according to claim 4 or claim substantially as hereinbefore described and/or exemplified.

23. A compound prepared by the process of claim 4 or claim

24. A pharmaceutical composition according to claim 5 or claim 16, substantially as hereinbefore described and/or exemplified. Use according to claim 8 or claim 19, substantially as hereinbefore described and/or exemplified.

P \OPER\Kbm\specis\12628860 specidoc

26. A method for the treatment of infectious diseases according to claim 9 or claim 20, substantially as Shereinbefore described and/or exemplified. DATED this 27 th day of June, 2005 00 Fujisawa Pharmaceutical Co., Ltd. and SWakunaga Pharmaceutical Co., Ltd. By DAVIES COLLISON CAVE Patent Attorneys for the Applicants