| Title of Invention | CEPHEM COMPOUNDS |
|---|---|
| Abstract | The present invention relates to a compound of the formula [I]: wherein R1 is lower alkyl, hydroxy(lower)alkyl or halo (lower)alkyl. and R1 is hydrogen amino protecting group, or R1 and R2 are bonded together and form lower alkylene or lower alkenylene; R3 is hydrogen or lower alkyl: R4 is ; R5 is carbony or protected carboxy; and R6 is amino or protected amino, or a pharmaceutically acceptable salt thereof a process for preparing a compound of the formula [I], and a pharmaceutical composition comprising a compound of the formula [I] in admixture with a pharmaceutically acceptable carrier. |
| Full Text | WO 2004/039814 PCT/JP2003/013684 DESCRIPTION CEPHEM COMPOUNDS TECHNICAL FIELD The present invention relates to new cephem 5 compounds and pharmaceutically acceptable salts thereof. More particularly, the present, invention relates to new cephem compounds and pharmaceutically acceptable salts thereof, which have antimicrobial activities, to processes for preparation thereof, to pharmaceutical 10 composition comprising the same, and to a method for treating infectious diseases in human being and animals. DISCLOSURE OF INVENTION One object of the present invention is to provide novel cephem compounds and pharmaceutically acceptable 15 salts thereof, which are highly active against a number of pathogenic microorganisms. Another object of the present invention is to provide processes for the preparation of said cephem compounds and salts thereof. 20 A further object of ths present invention is to provide a pharmaceutical composition comprising, as an active ingredient, said cephem compounds or their pharmaceutically acceptable salts. Still further object of the present invention is 25 to provide a method for treating infectious diseases caused by pathogenic microorganisms, which comprises administering said cephem compounds to infect&d human being or animals. The object cephem compounds of the present 30 invention are novel and can be represented by the following general formula [I]: 1 WO 2004/039814 PCT/JP2003/013684 wherein R1 is lower alkyl, hydroxy (lower) alkyl or halo(lower)alkyl, and 5 R2 is hydrogen or amino protecting group, or R1 and R2are bonded together and form lower alkylene or lower alkenylene; R3 is hydrogen or lower alkyl; R1 is wherein X is O or NH, 15 R7 is hydrogen, lower alkyl or amino protecting group, R3 is hydrogen or hydroxy, R9 is amino, mono or di(lower)alkylamino, protected amino, guanidino, protected 30 guanidino or saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or protected amino, k, m, n and q are independently 0 or 1, and 25 p is 0, 1, 2 or 3; R5 is carboxy or protected carboxy, and 2 WO 2004/039814 CT/JP2003/013684 R6 is amino or protected amino. As to the object compound (I), the following points are to be noted. That is , the object compound (I) include syn 5 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: 10 wherein R5 and R6 are each as defined above, and anti isomer (E form) means the other geometrical isomer having the partial structure represented by the following formula: 15 wherein R5 and R6 are each as defined above, and all of such geometrical isomer and mixture thereof are included within the scope ox this invention. In the present specification and claims, the partial structure of these geometrical isomers and 20 mixture thereof are represented for convenience' sake by the following formula: wherein R5 and R6 are each as defined above. Another point to be noted is that the pyrazolio 3 WO 2004/039814 PCT/JP2003/013684 moiety of the compound [I] can also exist in the tautomeric form, and such tautomeric equilibrium can be represented by the following formula. 5 wherein R1,R2,R3andR4 are each as defined above. Both of the above tautomeric isomers are included within the scope of the present invention, and in the present specification, and claims, however, the object compound [I] is represented for convenience' sake by one 10 expression of the pyrazolio group of the formula (A) . The cephem compound [I] of the present invention can be prepared by the following processes as illustrated, in the following. 15 4 WO 2004/039814 PCT/JP2003/013684 Process 1 or its reactive or its reactive or its reactive or its reactive derivative at the derivative at the derivative at the derivative at the amino group, carboxy group, or a salt thereof or a salt thereof 5 WO 2004/039814 PCT/JP2003/013684 Process 2 5 6 WO 2004/039814 PCT/JP2003/013684 7 WO 2004/039814 PCT/JP2003/013684 wherein R1,R2,R3,R4,R5,R6,R7,R8, A, k, m, n, p and 5 q are each as defined above, R10 is protected carboxy. Y is a leaving group, Z**** is an anion, R1a is protected hydroxy(lower)alkyl, 10 R1b is hydroxy(lower) alkyl, R9a is protected amino, protected guanidino or saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms substituted by protected amino, and R9b is amino, guanidino or saturated 3-to8-membered 15 heterocyclic group containing 1 to 4 nitrogen atoms substituted by amino. 8 WO 2004/039814 PCT/JP2003/013684 The starting compounds [II] and [VI] can be prepared by the following processes. 5 9 WO 2004/039814 PCT/JP2003/013684 wherein R1, R2, R3, R4, R5, R6,R10,YandZ** are each as 5 defined above, R11 is protected amino, R12 is protected carboxy, and R13 is amino protecting group or lower alkyl. The starting cotopounds [VII] and [XI] or salts 10 thereof can be prepared by the methods disclosed in the Preparations 3—6, 8-47 and 49-102 describ&d later or similar manners thereto. In the above and subsequent descriptions of this specification, suitable examples of the various 15 definitions ate explained in detail as follows. The term "lower" is used to mean a group having 1 10 WO 2004/039814 PCT/JP2003/013684 to 6, preferably 1to 4,carbon atoms, unless otherwise indicated. Suitable "lower alkyl" and "lower alkyl1' moiety in "hydro(lower)alkyl" , "protected hydroxy (lower) alkyl" , 5 "aryl (lower)alkyl", "halo (lower)alkyl" and "mono or di(lower)alkyl amino", include straight or branched alkyl having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert- butyl, pentyl, tart-pentyl and hexyl, In which more 10 preferred one is C1-C4 alkyl. Suitable "hydroxy(lower)alkyl" includes hydroxy (C1-C5) alkyl such as hydroxymethyl, 1- hydroxy ethyl, 2-hydroxyethyl,, 1-hydroxypropyl, 2- hydrdxypropyl, 3-hydroxypropyl, 4—hydroxybutyl, 5- 15 hydroxypentyl and 6 —hydroxyhexyl, in which more preferred one is hydroxy (C1-C4) 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 20 and fluorine. Preferred examples of "halo(lower)alkyl" include fluoromethyl, difluromethyl, trifluoromethyl, chloromethyl, bromomethyl, 2—fluoroethyl, 2 ,2- difluoroethyl, 2,2,2-trifluoroethyl, 2—chloroethyl, 2,2- dichloroethyl, 2,2,2—"trichloroethyl, 3—fluoropropyl and 25 2,2,3,3,3-pentafluoropropyl, in which more preferred one is halo (C1-C4) alkyl. Suitable "mono or di (lower)alkylamino" includes mono or di(C1-C6) alkylamino such as methylamino, dimethy1amino, ethylamino, diethylamino, N-ethyl—N— 30 methylamino, propylamino, butylamino and N-ethyl-N- propylamino, in which more preferred one is mono or di (C1-C4)alkyl amino. Suitable "lower alkylene" formed by R1 and R2 includes straight alkylene having 1 to 6, preferably 2 35 to 4 carbon atoms, such as methylene, ethylene, trilmethylene- and tetramethylene In which more preferred one is straight alkylene having 2 or 3 carbon atoms. Suitable "lower alkylene" formed by R1 and R2 11 WO 2004/039814 PCT/JP2003/013684 includes straight alkenylene having 2 to 6, preferably 2 to 4 carbon atoms, such as vinylene and propeoylene, in which more preferred one is straight alkylene having 2 or 3 carbon atoms. 5 Suitable "saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms" includes azetidinyl (e.g., l-azetidinyl and 3-azetidinyl), pyrrolidinyl (e.g., l-pyrro0lidinyl and 3-pyrrolidinyl), imidazolidinyl (e.g., 1-imidazolidinyl and 4- 10 imidazolidinyl), piperidinyl (e.g., 1-piperidinyl and 4- piperidinyl) and piperazinyl (e.g. , 1-piperazinyl) , in which, more preferred one is saturated 4- to 6-membered heterocyclic group containing 1 to 4 nitrogen atoms. The saturated 3-to 8-membered heterocyclic group 15 containing 1 to 4 nitrogen atoms is optionally substituted by amino or protected amino. Suitable examples of "saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substitutted by amino or protected amino" include 1- 20 azetidinyl, 3-amino—l-azetidinyl, 3-tert- butoxycarbonylamino-l-azetidinyl, 3-azetidinyl, 1- pyrrolidinyl, 3-amino-l-pyrrolidinyl, 3-tert- butoxycarbonylamino-l-pyrrolidinyl, 3-pyrrolidinyl, 1— piperidinyl, 4-piperidinyl and 1-piperazinyl. 25 Suitable "aryl" moiety in "aryl (lower) alkyl" includes C6-C12 aryl such as phenyl and naphthyl, in which, more preferred one is phenyl. Suitable "aryl(lower)alky1" includes mono-, di- or triphenyl (lower) alkyl such as benzyl, phenethyl, 30 benzhydryl and trityl. Suitable "lower alkanoyl" and "lower alkanoyl" moiety in "lower alkanoylamino" include straight or branched alkanoyl having 1 to 6 carbon atoms, such as formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, 35 isovaleryl, pivaloyl and hexanoyl, in which more preferred one is C1-C4 alkanoyl. Suitable "lower alkoxy" moiety in "lower alkoxycarbonyl" and "lower alkoxycarbonylamino" includes 12 WO 2004/039814 PCT/JP2003/013684 straight or branched alkoxy having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, tert- pentyloxy and hexyloxy, in which more preferred one is 5 C1-C4 alkoxy. Suitable "amino protecting group" in "protected amino" includes an acyl group as mentioned below, substituted or unsubstituted aryl(lower)alkylidene [e.g., benzylidene, hydroxybenzylidene, etc.], aryl(lower)alkyl 10 such as mono-, di—or tripaenyl (lower) alkyl) [e.g., benzyl, phenethyl, benzhydryl, trityl, etc.], and the like . Suitable "acyl" includes lower alkanoyl [e.g., formyl, acetyl, propionyl, hexanoyl, pivaloyl, etc.], 15 mono(or di or tri) halo (lower)alkanoyl [e.g-, chloroacetyl, trifluoroacetyl, etc.], lower alkoxycarbonyl [e.g., methoxycarbonyl, ethoxycarbonyl. tert-butoxycarbonyl, tert-pentyloxycarbonyl, hexyloxycarbonyl, etc.], carbamoyl, aroyl [e.g., benzoyl, 20 toluoyl, naphthoyl, etc, ] , aryl (lower) alkanoyl [e.g., phenylacetyl, phenylpropionyl, etc], aryloxycarbonyl (e.g., phenoxycarbonyl, naphthyloxycarbonyl, etc.], aryloxy (lower) alkanoyl [e.g., phenoxyacetyl, phenoxypropionyl, etc.], arylglyoxyloyl [e.g., 25 phenylglyoxyloyl, naphthylglyoxyloyl, etc.], aryl (lower) alkoxycarbonyl which optionally substituted by suitable substituent(s) [e.g., benzyloxycarbonyl, phenethyloxycarbonyl, p-nitrobenzyloxyearbonyl, etc.], and the like. 30 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 (C1-C6) alkyl, C1-C6 alkanoyl 35 and (C1-C6) alkoxycarbonyl. Preferable examples of "protected amino" include aryl(lower)alkylamino and acylamino, in which more preferred ones are aryl(lower)alkylamino, lower 13 WO 2004/039814 PCT/JP2003/013684 alkanoylamino and lower alkoxycarbonylamino, and particularly preferred ones are mono-, di- or triphenyl (C2-C5) alkylamino , C3-C5 alkanoylamino and (C2- C6)alkoxycarbonylamino. 5 Preferable examples of "protected guanidino" include acylguanidino (monoacylguanidino and diacylguanidino such as 2,3- bis [ (lower) alkoxycarbonyl]guanidino [e.g., 2 ,3-bis (tert- butoxycarbonyl) guanidinol , In which morepreferred one 10 is 2 , 3-bis [ (C1-C5) alkoxycarbonyl] gnanidino. 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 15 (e.g. , formyl. acetyl, propionyl,. hexanoyl, pivaloyl, etc.], mono (or di tri) halo (lower) alkanoyl, [e.g., chloroace tyl, trifluoroacetyl, etc.], lower alkoxycarbonyl, [e.g., methoxycarbonyl, ethoxycarbonyl, tert-butoxycarboyl, tert-pentyloxycarbonyl , 20 hexyloxycarbonyl, etc.] , carbamoyl, and -the like. Suitable "aryl (lower) alkyl" moiety in the "aryl (lower)alkyloxy" includes mono-, di- or triphenyl (lower) alkyl (e.g, , benzyl, phemethyl, benzydryl, trityl , etc.], and the like. 25 Suitable "protected carboxy" includes esterified carboxy and "the like, and concrete examples of estetrified carbaoxy include lower alkoxycarbonyl (e.g. , methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, 30 tert-butoxycarbonyl, pentyloxycarbonyl , hexyloxycarbonyl, l-cyclopropylethoxycarbonyl, etc. ] which may have suitable substituent (s) , for example, lower alkanoyloxy(lower)alkoxycarbonyl [e.g., acetoxymethoxycarbonyl, propionyloxymethosycarbonyl, 35 butyryloxymethoxycarbonyl, valeryloxymethoxycarbonyl, pivaloyloxymethoxycarbonyl, 1-acetoxyethoxycarbonyl, 1- propionyloxyethoxycarbonyl, 2 -propionyloxy thoxycarbo ny1, hexanoyloxymethoxycarbonyl, etc.], lower 14 WO 2004/039814 PCT/JP2003/013684 alkanesulfonyl (lower) alkoxycarbonyl, [e.g., 2- mesylethoxycarbonyl, etc, ] or mono (or di or tri) halo (lower) alkoxycarbonyl [e.g., 2- iodoethoxycarbony1, 2,2,2-trichloroethoxycarbonyl, 5 etc.I; lower alkenyloxycarbonyl [e.g., vinyloxycarbonyl, allyloxycarbonyl, etc.]; lower alkynyloxycarbonyl [e.g-, ethynyloxycarbonyl, propynyloxycarbonyl, etc.]; aryl(lower)alkoxycarbonyl which may have suitable substituent(s) [e.g. , benzyloxycarbony1, 4- 10 methoxybenzyloxycarbonyl, 4—nitrobezyloxycarbonyl, phenethyloxycarbonyl, trityloxycarbonyl, benzhydryloxycarbonyl, bis (methoxyphenyl) methoxycarbonyl, 3 ,4-dimethoxybenzyloxycarbonyl, 4-hydroxy-3 , 5-di-tert- butylbenzyloxycarbonyl, etc.]; aryloxycarbonyl which may 15 have suitable substituent(s) [e.g., phenoxycarbonyl, 4- chlorophenoxycarbonyl, tolyoxycarbonyl, 4-tert- butylphenoxycarbonyl , xylyloxycarbonyl, mesityloxycarbony1, cumanyloxycarbonyl, etc.]; and the like. 20 Preferable examples of "protected carboxy" include lower alkoxycarbonyl and aryl (lower) alkoxycarbonyl which may have suitable substituent(s) , in which more preferred one is (C1-C6) alkoxycarbanyl. Suitable "leaving group" includes halogen, [e.g., 35 chlorine, bromine, iodine, etc.] or acyloxy such as arylsulfonyloxy [e.g., benzenesulfonyloxy, tosyloxy, etc.], lower alkylsulfonyloxy [e.g., mesyloxy, etc.], lower alkanoyloxy [e.g., acetyloxy, propionyloxy, etc.], and the like. 30 Suitable "anion" includes formate, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate, toluenesulfonate, chloride, bromide, iodide, sulfate, hydrogen sulfate, phosphate, and the like. 35 Suitable pharmaceutically acceptable salts of the object compound [I] 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 15 WO 2004/039814 PCT/JP2003/013684 example, an alkali metal salt [e.g., sodium salt, potassium salt, etc.], an alkaline earth metal salt, [e.g., calcium salt, magnesium salt, etc] , an ammonium salt; a salt with an organic base, for example, an 5 organic amine salt [e.g., trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N,N'—dibenzylethylenediamine salt, etc.]; an inorganic acid addition salt [e.g, 10 hydrochloride, hydrobromide, sulfate, hydrogen sulfate, phosphate, etc,]; an organic carboxylic ot sulfonic acid addition salt [e.g., formate, acetate, trifluoroacetate, maleate, tartrate, citrate, fumarate, methanesulfonate, benzenesulfonate, toluenesulfoaate, etc.]; and a salt 15 with a basic or acidic amino acid [e.g., arginine, aspartic acid, glutamic acid, etc.]. The preferred embodiments of the cepkem compound of the present invention represented by the general formula [I] are as follows. 20 (1) The compound of the formula [I] wherein R1 is lower alkyl or hydroxy (lower) alky1,and R2 is hydrogen or amino protecting group, or R1 and R2 are bonded together and form lower alkylene; R3 is hydrogen; 30 p is 0r,1 or 2, or a pharmaceutically acceptable salt thereof. 12) The compound of (1) above wherein R3 is hydrogen, or a pharmaceutically acceptable salt thereof. (3) The compound of the formula [I] wherein 35 R1 is lower alkyl, hydroxy (lower) alkey1 or halo(lower)alkyl, and R2 is hydrogen, aryl(lower)alkyl or acyl, or 16 WO 2004/039814 PCT/JP2003/013684 R1 and R2 are bended together and form lower alkylene or lower alkenylene; R5 is carboxy or esterified carboxy; R6 is amino or acylamino; 5 R7 is hydrogen, lower alkyl or acyl; and R9 is amino, mono or di (lower) alkylamino , acylamino, guanidino, acylguanidino or saturated 3- to 8- membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or 10 acylamino, or a pharmaceutically acceptable salt thereof. (4) The compound of (3) above wherein R1 is lower alkyl or hydroxy (lower) akkyl, and R2 is hydrogen, aryl(lower)alkyl or acyl, or 15 R1 and R2 are bonded together and form lower alkylene; R5 is carboxy or esterified carboxy; R6 is amino or acylamino; R7 is hydrogen or acyl; and R9 is amino or acylannino, 20 or a pharmdceutically acceptable salt thereof. (5) The compound of (4) above wherein R1 is lower alkyl or hydroxy (lower) alky1, and R2 is hydrogen, aryl(lower)alkyl, lower alkanoyl or lower alkoxycarbonyl, or 25 R1 and R2 are bonded together and form lower alkylene; R5 is carboxy or lower alkoxycarbonyl; R6 is amino, lower alkanoylamino or lower alkoxycarbonylamino; R7 is hydrogen, lower alkanoyl or lower alkoxycarbonyl; 30 and R3 is amino, lower alkanoylamino or lower alkoxycarbonylamino, or a pharmaceutically acceptable salt thereof. (6) The compound of (5) above wherein 35 R1 is C1—C6 alkyl or hydroxy (C1-C6) alkyl, and R2 is hydrogen, mono-, di- or triphenyl (C1-C6) alkyl, C1-C6 alkanoyl or (C1-C6) alkoxycarbonyl, or R1 and R2 are bonded together and form C1-C6 alkylene; 17 WO 2004/039814 PCT/JP2003/013684 R5 is carboxy or (C1-C6) alkoxycarbonyl ; R6 is amino, C1-C6 alkanoylamino or (C1-C6) alkoxycarbonylamino ; R7 is hydrogen, C1-C6 alkanoyl or (C1-C6) alkoxycarbonyl; 5 and R9 is amino, C1-C6 alkanoylamino or (C1-C6) alkoxycarbonylamino , or a pharmaceutically acceptable salt thereof. (7) The compound of (5) above wherein 10 R1 is lower alkyl or hydroxy(lower)alkyl , and R2 is hydrogen, or R1 and R2 are bonded together and form lower alkylene; R5 is carboxy; R6 ia amino; 15 R7 is hydrogen or lower alkanoyl; and R9 is amino, or a. pharmaceutically acceptable salt thereof. (8) The compound of (7) above wherein, R1 is C1-C6 alkyl or hydroxy (C1-C6) alkyl , and 20 R2 is hydrogen, or R1 and R2 are bonded together and form C1-C6 aIkylene; R5 is carboxy; R6 is amino; R7 is hydrogen or C2-C6 alkanoyl; and 25 R9 is amino, or a pharmacetically acceptable salt thereof, (9) The compound of the formula [I] wherein R4 is selected from the group consisting of 18 WO 2004/039814 PCT/JP2003/013684 wherein R7, A, m, p and q are each as defined above in 5 the formula [I], R14 is amino, mono or di(lower)alkylamino or protected amino, Rl5 is guanidino or protected guanidino, and R16 is saturated 3- to 8-membered heterocyclic group 10 containing 1 to 4 nitrogen atoms optionally substituted by amino or protected amino, or a pharmaceutically acceptably salt thereof. (10) The compound of the formula [I] wherein R4 is selected from the group consisting of 19 WO 2004/039814 PCT/JP2003/013684 wherein p is 0, I or 2, q is 0 or 1, 5 R7 is hydrogen or amino protecting group, and R9 is amino or protected amino, or a pharmaceutically acceptable salt thereof. (11) The compound of (10) above wherein R7 is hydrogen, lower alkanoyl or lower alKoxycarbonyl; 10 and R3 is amino, lower alkanoylamino or lower alkoxycarbanylamino, or a pharmaceutically acceptable salt thereof. (12) The compound of (II) above wherein 15 R7 is hydrogen. C1-C6 alkanoyl or (C1-C6) alkoxycarbonyl; and R9 is amino, C1-C6 alkanoylamino or (C1-C6) alkoxycarbonylamino, or a pharmaceutically acceptable salt thereof. 20 (l3) The compound of (11) above wherein R7 is hydrogen or lower alkanoyl; and R9 is amino, or a pharmaceutically acceptable salt thereof. (14) The compound of (13) above wherein 25 R7 is hydrogen or C1-C5 alkanoyl; and R9 is amino, or a pharmaceutically acceptable salt thereof. The processes for preparing the object compound of 30 the present invention are explained in detail in the following. Process 1 The compound [I] or a salt thereof can be prepared by reacting the compound [II] or its reactive derivative 35 at the amino group, or a salt thereof with the compound [III] or its reactive derivative at the carboxy group, 20 WO 2004/039814 PCT/JP2003/013684 or a salt thereof. Suitable reactive derivative at the amino group of the compound [II] includes Schiff's base type amino or its tautomeric euamifle type isomer formed by the 5 reaction of the compound [II] with a carbonyl compound sucn as aldehyde, xetone and the like; a silyl derivative formed by the reaction of the compound [II] with a silyl compound such as bis(trimethylsilyl)acetamide, 10 mono (trimethylsilyl) acetamide [e.g., N- (trimethylsilyl) acetamide] , bis (trimethylsilyl) urea and the like; a derivative formed by the reaction of the compound [II] with phosphorus trichloride or phosgene. Suitable salts of the compound [II] and its 15 reactive derivative can be referred to the ones as exemplified for the compound [1]. 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. 20 A suitable example of the reactive derivatives may be an acid chloride; an acid azide; a. milted acid anhydride with an acid such as substituted phosphoric acid [e.g., dialkylphosphoric acid, phenylphosphoric a.cid, diphenylphosphoric acid, dibenzylphosphoric acid, 25 halogenated phosphoric acid, etc.], dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, sulfuric acid, alkanesulfonic acid [e.g, methanesulfonic acid, etc.], aliphatic carboxylic acid [e.g., acetic acid, propionic acid, butyric acid, isobutyric acid, pivalic acid, 30 pentanoic acid, isopentanoic acid, 2-ethylbutyric acid, trichloroacetic acid, etc.] and aromatic carboxylic acid [e.g., benzoic acid, etc,]; a symmetrical acid anhydride; an activated amide with imidazole, 4- substituted imidazole, dimethylpyrazole, triazole or 35 tetrazole; an activated ester [e.g., cyanomethyl ester, methoxymethyl ester, dimethyliminoimethyl (CH3)2N+=CH-] ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, 21 WO 2004/039814 PCT/JP2003/013684 pentachlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenyl thioester, p-nitrophenyl thioester, p—cresyl thioester, carboxymethyl thioester, pyranyl ester, pyridyl ester, piperidyl ester, 8- 5 quinolyl thioester, etc.]; or an ester with an N-hydroxy compound [e.g.,N,N-dimethylhydroxylamine, 1-hydroxy-2- (1H)-pyridone, N-hydroxysuccinimide, N- hydroxyphthalimide, N-hydroxy-lH-benzotriazole, etc. ] . These reactive derivatives can optionally be selected 10 from them according to the kind of the compound [III] to be used. Suitably salts of the compound [III] and its reactive derivative can be referred to the ones as exemplified for the compound [I]. 15 The reaction is usually carried out in a conventional solvent such as water, alcohol [e.g., methanol, ethanol, etc.], acetone , dioxane , acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N ,N-dimethylformaraide , 20 pyridine or any other organic solvent which does not adversely affect the reaction. Thsae 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 25 preferably carried out in the presence of a conventional condensing agent such as N,N' dicyclohexylcarbodiimide; N-cyclohexyl-N'-morpholinoethylcarbodimide; N— cyclohexyl-N'—(4-diethylaminocyclohexyl) carbodiimide; N,N'-diethylcarbodiimide; N,N'-diisopropylcarbodiimide; 30 N-ethyl-N'-(3—dimethylaminopropyl)carbodiimide; N,N'— carbonyl-bis- (2—methylimidazole); pentamethyleneketane- N-cyclohexylimine; diphenylketene--N-cycloh.Qxylimine; ethoxyacetylene; I-alkoxy-I-chloroethylene; trialkyl phosphite; ethyl polyphosphate; isopropyl polyphosphate; 35 phosphorus oxychloride (phosphoryl chloride) ; phosphorus trichloride: thionyl chloride; oxalyl chloride; lower alkyl haloformate [e.g., ethyl chloroformate, isopropyl chloroformate, etc.], triphenylphosphine; 2-ethyl-7- 22 WO 2004/039814 PCT/JP2003/013684 hydroxybenzisoxazolium salt; 2-ethyl-5- (m- sulfophenyl)isonxazolium hydroxide intramolecular salt; l-(p-chlorobenzenesulfonyloxy) -6-chloro-lH- benzotriazole; so-called Vilsmeier reagent prepared by 5 the reaction of N,N-dimethylformamide with thionyl 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 bass such, as an 10 alkali metal bicarbonate, tri(lower)alkylamine, pyridine, N- (lower)alkylmorpholine, N,N—di(lower)aIkylbenzylamine, and the like. The reaction temperature is not critical, and the reaction is usually carried out under cooling to warming. 15 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. 20 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. 25 Suitable base includes an inorganic base and an organic base such as an alkali metal [e.g., sodium, potassium, etc.], an alkaline earth metal [e.g., magnesium, calcium, etc.], the hydroxide or carbonate or hydrogencarbonate thereof, trialkylamine [e.g., 30 trimethylamine, triethylamine, etc.], picoline, 1,5- 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 [e.g., 35 formic acid, acetic acid, propionic acid, trichloroacetic acid, trifluoroacetic acid, etc.], and an inorganic acid [e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, hydrogen chloride, hydrogen bromide, 23 WO 2004/039814 PCT/JP2003/013684 etc.]. The elimination using Lewis acid such as trihaloacetic acid [e.g., trichloroacetic acid, trifluoroacetic acid, etc.], and the like is preferably 5 carried out in the presence of cation trapping agents [e.g., anisole, phenol, etc.]. The reaction is usually carried out in a solvent such, as water, alcohol [e.g. , methanol, ethanol, etc:.] , methylene chloride, tetrahydrofuran, a mixture thereof 10 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 cut under cooling to warming. 15 Process 3-(i) The compound [VIII] or a salt thereof can be prepared by reacting the compound [VII or a salt thereof with the compound [VII] or a salt thereof. Suitable salt of the Compounds [VI], [VII] and 20 [VIII] can be referred to the ones as exemplified for the compound [I] . The present reaction may be carried out in a solvent such as water, phosphate buffer, acetone, chloroform, acetonitrile nitrobenzene, methylene 25 chloride, ethylene chloride, formamide, N ,N- dimethylformamide, methanol, ethanol, diethy1 ether, tetrahydrofuran, dimethyl sulfoxide, or any other organic solvent which does not adversely affect the reaction, preferably in ones having strong polarities. 30 Among the solvent, hydrophilic solvents may be used in a mixture with water. When the compound [VII is liquid, it can also be used as a solvent. The reaction is preferably conducted in the presence of a base, for example,- an inorganic base such 35 as alkali metal hydroxide, alkali metal carbonate, alkali methyl hydrogencarbonate , an organic basa such as trialkylamine, and the like. The reaction temperature is not critical, and the 24 WO 2004/039814 PCT/JP2003/013684 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 [e,g., sodium iodide, potassium iodide, etc.], 5 alkali metal thiocyanate [e.g., sodium thiocyanate, potassium thiocyanate, etc.], and the like. Anion Z**** may be one derived from a leaving group Y, and it may be converted to other anion by a conventional method. 10 Process 3-(ii) The compound [I] or a salt thereof can be prepared by subjecting the compound [VIII] or a salt thereof to elimination reaction of the carboxy protecting group. Elimination reaction is carried out in similar 15 manner to the reaction in the aforementioned Process 2, and therefore the reagents to be used and reaction conditions (e.g., solvent, reaction temperature, etc.) can be referred to those of Process 2. Process 4 20 The compound [Id] or a salt thereof can be prepared by subjecting the compound [Ic] or a salt thereof to elimination reaction of the hydroxy protecting group. Suitable method of this elimination reaction 25 includes conventional one such as hydrolysis, reduction and the like, (i) For hydrolysis; The hydrolysis is preferably carried out in the presence of a base or an acid including Lewis acid. 30 Suitable base includes an inorganic base and an organic base such as an alkali metal [e.g., sodium, potassium, etc.], an alkaline earth metal [e.g., magnesium, calcium, etc.], the hydroxide or carbonate or hydrogencarbonate thereof, trialkylamine [e,g., 35 trimethylamine, triethylamine, etc.], picoline, 1,5— 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. 25 WO 2004/039814 PCT/JP2003/013684 Suitable acid includes an organic acid [e.g., formic acid, acetic acid, propionic acid, trichloroacetic acid, trifluoroacetic acid, stc], and an inorganic acid [e.g., hydrochloric acid, hydrobromic 5 acid, sulfuric acid, hydrogen chloride, hydrogen bromide, etc.]. The elimination using Lewis acid such as trihaloacetic acid [e.g., tri-chlroacetic acid, trifluoroacetic acid, etc] and the like is preferably 10 carried out in the presence of cation trapping agents [e.g., anisole, phenol, etc.]. The reaction is usually carried out in a solvent such as water, alcohol [e.g., methanol , ethanol, etc.] , methylene chloride, t^trahydrofuran, a mixture thereof 15 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. 20 (ii) For reduction: Reduction is carried out in a conventional manner, including chemical reduction and catalytic reduction. Suitable reducing reagents to be used in chemical reduction are a combination of a metal [e.g., tin, zinc, 25 iron, etc] or metallic compound [e.g. , chromium chloride, chromium acetate, etc.] and an organic acid or inorganic acid [e.g., formic acid, acetic acid, propionic acid, trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid, hydrobromic acid, etc.]. 30 Suitable catalysts to be used in catalytic reduction are conventional ones such as platinum catalysts [e.g. , platinum plate, spongy platinum, platinum black, colloidal platinum, platinum oxide, platinum wire, etc.], palladium catalysts [e.g., spongy 35 palladium, palladium black, palladium oxide, palladium on carbon, colloidal palladium, palladium on barium sulfate, palladium on barium carbonate, etc], nickel catalysts [e.g., reduced nickel, nickel oxide, Raney 26 WO 2004/039814 PCT/JP2003/013684 nickel, etc], cobalt catalysts [e.g., reduced cobalt, Raney cobalt, etc.], iron catalysts [e.g., reduced iron, Raney iron, etc.],copper catalysts [e.g., reduced copper, Raney copper, ullman copper, etc.] and the like. 5 The reduction is usually carried out in a conventional solvent which does not adversely influence the reaction such as water, methanol, ethanol, propanol, N,N-dimethylformamide or a mixture thereof. Additionally, in case that the above—mentioned 10 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 reduction may be the above-mentioned solvent, and other conventional solvent such as diethyl ether, 15 dioxane, 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. When R6 is protected amino, the amino protecting 20 group in R6 can be eliminated by a conventional method such as hydrolysis. Processes A and B for the preparation of the starting compounds are explained in detail in the following. 25 Process A-(i) The compormd [XII] or a salt thereof can be prepared by reacting the compound [X] or a salt thereof with the compound [XI] or a salt thereof. This reaction, can be carried out in a similar 30 manner to the reaction in the aforementioned Process 3— (i), and therefore the reagents to be used and reaction conditions [e.g., solvent, reaction temperature, etc.) can be referred to those of Process 3- (i) . Process A-(ii) 35 The compound [II] or a salt thereof can be prepared by subjecting the compound [XII] or a salt thereof to elimination reaction of the amino protecting groups in R11 and R13 and the carboxy protecting group in 27 WO 2004/039814 PCT/JP2003/013684 R12 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 5 conditions (e.g., solvent, reaction temperature, etc.) can be referred to those of Process 2. Process B The compound [VI] or a salt thereof can be prepared by reacting the compound [XIII) or its reactive 10 derivative at the amino group, or a salt thereof with the compound [XIV] 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, 15 and therefore the reagents to be used and reaction conditions (e.g., solvent, reaction temperature, etc.) can be referred to those of Process_l. The compounds obtained by the above processes can be isolated and purified by a conventional method such 20 as pulverization, recrystallization, column chromatography, reprecipitation, and the like. It is to be noted that the compound [I] and other compounds may include one or more stereoisomer(a) such as optical isomer(s) and geometrical isomer(s) due to 25 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 object compounds [I] and pharmaceutically acceptable salts thereof include solvates [e.g., 30 enclosure compounds (e.g., hydrate, etc.)]. The object compound [I] and pharmaceutically acceptable salts thereof are novel and exhibit high antimicrobial activity, inhibiting the growth of a wide variety of pathogenic microorganisms including Gram— 35 positive and Gram-negative microorganisms and are useful as antimicrobial agents. Now in order to show the utility of the object compound [I] the test data on MIC (minimal inhibitory 28 WO 2004/039814 PCT/JP2003/013684 concentration) of a representative compound of this invention are shown in the following. Test method In vitro antibacterial activity was determined by 5 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 viabls cells per ml) was streaked on heart infusion agar (Hl-agar) containing 10 graded concentrations of representative -test compound, and tine minimal inhibitory concentration (MIC) was expressed in mg/ml after incubation at 37oC for 20 hours . Test compound Compound (a): 7- [ (Z) -2- (5-amino-l , 2 , 4-thiadiazol-3-yl) - 15 2- (l-carboxy-l-methylethoxyimino)acetamido]-3-[7-(3- aminopropionamido)-2,3-dihydro-5-(1H-imidazo[1,2- blpyrazolio)]methyl-3-cephem-4-carboxylate (Example 3) Compound (b): 7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)- 2- (l-carboxy-l-methylethoxyimino) acetamido] -3- [3-amino- 20 4- (3-aminopropionamido) -2-methyl-l-pyrazolio]methyl-3- cephem-4-carboxylate (Example 4) Compound (c): 7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)- 2—(1-carboxy-l-methylethoxyimino)acetamido]-3-[3-amino- 4— (aminoacetyl) amino-2-methyl-1- pyrazolio]methyl-3- 25 cephem-4-carboxylic acid hydrogen sulfate (Example 6) Compound (d) : 7- [(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)- 2—(1-carboxy-l-methylethoxyimino)acetamido]—3-(3-amino- 4-[3-(2-aminoethyl)ureido]-2-methyl-l-pyrazolio methyl- 3—cephem-4-carboxylic acid hydrogen sulfate (Example 7) 30 Compound (e): 7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-y1)- 2- (1-carboxy-l-methylethoxyimino) acetamido] -3- (3-amino- 4—guanidino-2—methyl-l-pyrazolio)methyl-3-cephem-4- carboxylic acid hydrogen sulfate (Example 11) Ceftazidime 35 29 WO 2004/039814 PCT/JP2003/013684 Test: results For therapeutic administration, the object 5 compound [I] 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 10 or inorganic solid or liquid excipient which is suitable for oral, parentetal 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 15 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, 20 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 [I] may vary from and also depend upon the age, conditions of the patient, 25 a kind of diseases, a kind of the compound (I) to be applied, etc. In general amounts between 1 mg and 4,0 00 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 mg of the object compounds [I] 30 of the present invention may be used in treating diseases infected by pathogenic microorganisms. The following Preparations and Examples are given 30 WO 2004/039814 PCT/JP2003/013684 for the purpose of illustrating the present invention in more detail. Preparation 1 To a solution of (Z) -2- (5-amino-l,2,4-thiadiazol- 5 3-y1) [(2-tert-butoxy-1,1-dimethy1-2- oxoethoxy) imino] ethanoic acid (5g) in a mixture of tetrahydrofuran (80 ml) and N,N-dimethylformaide (20 ml) was added a solution of sodium bis(trimethylsilyl)amide (8.33 g) in tetrahydrofuran (12 10 ml) , and the mixture was stirred for 15 minutes. To the reaction mixture was added a solution of di-tert-butyl dicarbonate (3.3 g) in tetrahydrofuran (20 ml) under ice-cooling, and the mixture was stirred under ice- cooling for 3 hours. To the reaction mixture was added 15 ethyl acetate, and the mixture was washed with 10% aqueous potassium hydrogen sulfate solution, and then washed with, a phosphate buffer (pH 6.86). The organic layer was separated, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The 20 residue was triturated with diisopropyl ether and dried in vacuo to give (Z)-2-(5—[(tert-butoxycarbonyl)amino]- 1,2,4-thiadiazol-3-Yl)[(2-tert-butoxy-l, l-dimethyl-2- oxoethoxy) imino] etlianoic acid (3.10 g) , IR(KBr) 3191.6. 2981.4, 1714.4, 1550.5, 1153.2, 1000.9 25 cm-1 1H-NMR(DMSO-d6) .37 (9H, s), 1.45 (6H,s), 1.50 (9H, S) , 12.7 (1H,s) ESI-MS: m/z=429(M-H) Preparation 2 30 A mixture of N ,N-dimethylformamide (0.640 ml) and phosphoryl chloride (0.781 ml ) was stirred at room temperature for 30 minutes . To the mixture were added tetrahydrofuran (4 ml) and (Z)-2-(5-[(tert- butoxycarboxyl)amino]-l,2,4-thiadiazol-3-yl [ (2-tert- 35 butoxy-1,1 -dimethyl—2-oxoethoxy) imino] ethanoic acid (3 g) at 4°C, and the reaction mixture was stirred at room temperature for 1 hour. Meanwhile, a mixture of benzhydryl 7-amino-3-chloromethyl-3-cephem-4- 31 WO 2004/039814 PCT/JP2003/013684 carboxylate hydrochloride (3 g) and N- (trimethylsilyl)acetamide (8.72 g) in tetrahydrofuran (15 ml) was warmed to make a clear solution. The solution was then, cooled to -20°C and added to the 5 activated acid solution obtained above. The reaction mixture was stirred at a temper attire of —10°C to 0°C for 1 hour and poured into a mixture of ethyl acetate and water. The aqueous layer was separated, and the organic layer was washed with brine, dried over anhydrous 10 magnesium sulfate and filtered. The filtrate was concentrated in vacuo and purified by column chromatography on silica gel eluting with hexane/ethyl acetate (3:2) to give benzhydryl 7(Z)-2-(5-tert- butoxycarboxYlamino—1 ,2, 4-thiadiazol-3—yl) -2- (1-tert- 15 butoxycarbonyl-1-methylethoxylmino) acetamido] -3- chloromethyl-3-cephem-4-carboxylate (4.79 g) . rR(KBr 2981.4, 1793.5, 1720.2, 1524.8, 1371.1, 1247.7, 1151.3 cm-1 1H-NMR(DMSO-d6) 1.39 (6H, s), 1-48 (3H, s), 1.5 0 (6H, 20 s), 3.58 (1H, d, J-l8.3Hz), 3.76 (1H, d, J=l8.3Hz), 4.44 (2H, s), 5.29 (1H, d, J-5.0Hz), 6.01 (lH, dd, J=8.6, 5.OHz), 6.97 (1H, s) , 7.2-7.6 (10H, m) , 9.65 (1H,d, J=5.OHz), 12.7 (1H, s) ESI-MS: m/z-849(M+Na) 25 preparation 3 To a solution of 5-amino-l-methylpyrazole (5 g) in ethanol (5O ml) was added isoamyl nitrite (6.92 ml) , and then 20% hydrochloric acid (5 drops) was added at 4°C. The reaction mixture was refluxed for 2 hours and cooled 30 to room temperature . To the reaction mixture was added diisopxopyl 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-nitirosopyrazole (3.53 g) , 35 1H-NMR(DMSO-d6) 3.51 (3H, s), 8.07 (2H, brs) , 8.51 (1H, s) APCI-MS; m/z=l27(M+H) Preparation 4 32 WO 2004/039814 PCT/JP2003/013684 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 5 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 sulfuric acid salt 10 (1. 71 g) . 1H-NMR(DMSO-d6) 3.54 (3H, s), 7.19 (1H, s) ESI-M:- m/z=113 (M+H) Preparation 5 To a suspension of 1,1'-carbonyldiimidazole (9.73 15 g) in dehydrated chloroform (72 ml) was added tert-butyl N-(2=aminoethyl)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 (l4.22 g) and 4,5-diamino-l- 20 methylpyrazole sulfuric acid salt (10.51 g) , and the mixture was stirred at 50oC for 15 hours. The insoluble materials were removed by filtration. To the filtrate were added chloroform (200 ml) and 5% aqueous sodium hydrogen carbonate solution (100 ml). The organic layer 25 was separated, and the aqueous layer was extracted with a mixed solvent of chloroform and methanol(4:1). The organic layers were combined, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with ethyl acetate and dried 30 in vacuo to give 5-amino-4- (3- (2- [(tert- butoxycarbonyl)amino]ethyl)ureido)-1—methylpyrazole (14.0 g) as a solid. 1H-NMR(DMSO-d6) 1.38 (9H, s), 2.36-2.98 (2H, m) , 3.03- 3.07 (2H, m), 3.50 (3H, s), 4.81 (2H, br), 5.92 (lH, br), 35 6.80 (1H, br) , 6.96 (1H, s), 7.18 (1H, br) Example 1 To a solution of benzhydryl 7b- [ (Z)-2- (5-tert- butoxycarbonylamino-1,2,4-thiadiazol-3-yl) -2-(l-tert- 33 WO 2004/039814 PCT/JP2003/013684 butoxycarbonyl-l-methylethoxyimino) acetamido] -3- chloromethyl-3-cephem-4-carboxylate (500 mg) in N,N- dimethylformamide (1.0 ml) was added sodium iodide (10 0 mg) , and the mixture was stirred at room temperature for 5 3 0 minutes. To the reaction mixture was added a Solution of 5-amino-4—(3—(2—[(tert- butoxycarbonyl)amino]ethyl]ureido)-1-methylpyrazole (216 mg) in N,N-dimethylformamide (1.0 ml) . The whole mixture was stirred at 32oC for 4 hours. To the 10 resulting reaction mixture were added ethtyl acetate (50 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed with 10% acqueous sodium trifluoroacetate solution and brine, dried over anhydrous sodium, sulfate and filtered. The filtrate was 25 concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (75 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene chloride (1.8 ml) were added anisole (0.6 ml) 20 and trifluoroacetic acid (1.2 ml). The resulting solution was stirred at room temperature for 4 hours and poured into diisopropyl ether 180 ml) . The resulting precipitate was collected by filtration and dried in vacuo to give a crude product (380 mg), which was 25 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 with concentrated hydrochloric acid and 30 chromatographed on Diaion® 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 7— [ (Z) — 2— (5—amino— 1, 2 , 4—thiadiazol- 3-yl)—2-(1—carboxy-1-methylethoxyimino)acetamido]-3-(3- 35 amino-4-[3-(2-aminoethyl)ureido]-2-methyl-l- pyrazolio)methyl-3-cephem—4-carboxylate (21 mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s), 3.12 (2H, t, 34 WO 2004/039814 PCT/JP2003/013684 J=5.7HZ), 3.22 (1H, d, J=17.9Hz), 3.49 (1H, d, J-17.9Hz), 3.46 (2H, t, J=5.7Hz), 3.71 (3H, s), 4.95 (1H, d, J-15.6Hz) , 5.15 (1H, d, J=l5.6Hz), 5.25 (1H, d, J=4.6Hz), 5.84 (1H, d, J=4.6Hz), 7.69 (1H, s) 5 Preparation 6 To a solution of 5-amino-4-(3-[2-[(tert- butoxycaxbonyl] amino] ethyl)urido) -l-methylpyrazole (597 mg) and triethylamine (243 mg) in methylene chloride (10 ml) was added triphenylmethyl chloride (669 mg) , and the 10 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, 15 filtered and cocentrated in vacua. The residue was triturated with ethyl acetate to give 4-(3-(2-f(tert- butoxycarbonyl amino] ethyl)) l-methyl-5- triphenylmethylaminopyrazole (640 mg) as a solid. 1H-NMR(DMSO-d6) 1.38 (9H, s), 2.70 (3H, s), 2.94-2.96 20 (2H, m) , 2.99-3.01 (2H, m) , 5.68 (1H, brs) , 5.96 (lH, br) , 6.78 (1H, br) , 6.85 (H, br) , 7.00 (1H, s), 7.13- 7.15 (6H, m), 7.24-7.28 (9H, m) Preparation 7 To a solution of benzhydryl 7- [ (Z) -2- (5-amino- 25 l,2,4-thiadiazol-3-yl)-2- (l-tert-butoxycarbonyl-l- methylethoxyimino) acetamido)-3-chloromethyl-3-cephem-4- carboxylate (60 g) in toluene (600 ml) were added a solution of sodium iodide (61.8 g) in 0.05 mol phosphate buffer (pH 7, 50 0 ml) and tricaprylylmethylammonium 30 chloride (6.67 g). The mixture was stirred at room temperature for 15 hours . The reaction mixture was added to a mixture of ethyl acetate and water. The organic layer was washed with water and brine, and then dried over magnesium sulfate. The magnesium sulfate was 35 filtered off, and the filtrate was evaporated to 255 g under reduced pressure. The concentrate was poured into diisopropyl ether (2 L) . The resulting precipitate was collected by filtration and dried to give benzydryl 7b- 35 WO 2004/039814 PCT/JP2003/013684 [[Z)-2-(5-amino-l,2,4-thiadiazol-3-yl) -2-(1-tert- butoxycarbonyl—1-methylethoxyimino) acetamido] -3- iodomethyl-3-cephem-4-carboxYlai:e (59.4 g) , 1H-NMR(DMSO-d6) 1.39 (9H, s), 1.46 (6H, s), 3.57 and 5 3.87 (2H, ABq, J=l8.OHz), 3.76 (3H, S), 4.30 (2H, bs), 5.25 (1H, d, J=4.9Hz), 5.94 flH, dd, J=4.9, 8.7Hz), 6,95 (1H, bs) , 7.15-7.60 (10H, m) , 8.17 (2H, bs) , 9.53 (1H, d, J=8.7Hz) Example 2 10 To a solution of benzhydryl 7 [ (Z) -2- (5-amino- 1,2,4-thiadiazol-3-yl)-2-(l-tert-butoxycarbonyl-l- methylethoxyimino) acetamido] — 3-iodomethyl-3—cephem-4- carboxylate (810 mg) in N,N-dimethylformamide (2.4 ml) was added N— (trimethylsilyl) acetamide (656 mg) , and the 15 mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added a solution of 4-(3-(2- [(tert-butoxycarbonyl)amino]ethyl)ureido) -l-methyl-5- triphenylmethylaminopyrazole (640 mg) in mettylen chloride (1O ml). The whole mixture was stirred at room 20 temperature for 26 hours. To the resulting reaction mixture were added ethyl acetate (50 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed with 10% aqueous sodium trifluoroacetate solution and brine, dried over 25 anhydrous sodium sulfatet and filtered. The filtrate was concentrated to about: 5 ml vacuo. The concentrate was poured into diisopropyl ether (80 ml) , and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in 30 methylene chloride (2.380 ml) were added anisole (0.79 ml) and trifluoroacetic acid (1.58 ml) . The resulting solution was stirred at room temperature for 4 hours and poured into diisopropyl ether (80 ml) . The resulting precipitate was collected by filtration and dried in 35 vacuo to give a crude product (635 mg), which was purified by preparative HPLC utilizing 0DS column. The eluate containing a desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to 36 WO 2004/039814 PCT/JP2003/013684 about pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2—propanol. The eluate was concentrated to about 30 ml in vacuo and 5 lyophilized to give 7- [ (Z) -2-(5-amino-l ,2,4-thiadiazol- 3-y1) -2- (1-carboxy-l -methylethoxyimino) acetamido ] —3 — (3- amino-4- [3- (2-aminoethyl) ureido] -2-methyl-l pyrazolio )methyl-3-cephem-4—carboxylate (54 mg) as an amorphous solid. 10 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s) , 3.12 (2H, t, J=5.7Hz), 3.22 (1H, d, J=17.9Hz), 3.49 (1H, d, J=17.9Hz), 3.45 (2H, t, J=5.7Hz), 3.71 (3H, s), 4.95 (1H, d, J=l5.6Hz) 5.15 (1H,d, J=15.6Hz), 5.25 (lH, d, J=4.6Hz), 5.84 (1H, d, J=4.6Hz), 7.89 (1H, s) 15 Preparation 8 To a solution of 2,3-dihydro-lH-imidazo[1,2- b)pyrazole (120 g) in sulfuric acid (500 ml) was added potassium nitrate (111 g) under ice-cooling. The mixture was stirred at room temperature for 48 hours. 20 The reaction mixture was added to ice (2.0 kg). The crystalline residue was collected by filtration and dried invacuo to give 7-nitro-2,3-dihydro-1H- imidazo[1,2-b]pyrazole (132 g) as a solid. 1H-NMR(DMSO-d6) 4.05-4.03 (2H, m) , 4.17-4.20 (2H, m), 25 7.82 (1H, s), 7.97 (1H, br) Preparation 9 A suspension of 7-nitro-2,3-dihydro-lH— imidazo [1,2-b] pyrazole (97 g) in a mixed solvent of sulfuric acid (34 ml) and water (2000 ml) was treated 30 with 10% palladium on carbon (10 g) under a hydrogen atmosphere at room temperature for 4 days. After the catalyst was filtered off, the filtrate was concentrated in vacuo. The residue was triturated with methanol and dried in vacuo to give 7-amin0-2,3-dihydro-lH- 35 imidazol [1,2-b]pyrasole sulfuiric acid salt (90 , 2 g) as a solid. 1H-NMR(DMSO-d6) 3.87-3.90 (2H, m0 , 4.07-4.10 (2H, m) , 7.28 (lH, s) 37 WO 2004/039814 PCT/JP2003/013684 Preparation_l_0 To a solution of 7-amino-2,3-dihydro-lH- imidazo[,2-b]pyraols sulfuric acid salt (22 g) and N-ethyldiisopropylamine (2.84 g) in methylene chloride 5 (70 ml) was added N- [3- (tert-butoxycarbonylamino) - propionyloxy]succinimide (3.15 g) . The mixture was stirred at room temperature for 4 Lours. The reaction mixture was washed with saturated aqueous sodium hydrogen carbonate solution, and the organic layer was 10 dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The oily residue was purified by calumn chromatography on silica gel eluting with 5% methanol/chloroform to give 7— [3- (tert- butoxycarboxylamino) propionyl ] amino-2 , 3-dihydro-lH- 15 imidazo[l,2-b]pyrazole (2.2 g) as a solid, 1H-NMR(CDCl3) 1.44 (9H, s), 2.52 (2H, t, J-6.0Hz), 3.36-3.47 (2H, m) , 3. 96 (2H, t, J-8,2H2), 4 .18 (2H, t) J=8.2Hz), 5.16 (1H, br), 7.16 (1H, s), 7.90 (lH, br) Example 3 20 7b-[ (Z) -2-(5-Amino-l,2,4-thiadiazol-3-yl) -2- (l- carboxy-l-methylethoxyimino) acetamido] -3- [ 7- [3- aminopropionamido) -2 , 3-dihydro-5- (1H-imidazo [1,2- b]pyrazolio ]methyl-3-cephem-4carboxylate The title compound was obtained from benzhydryl 25 7 b[(Z)-2-(5-tert-butoxycarbonylamino-1,2,4-thiadiazol- 3-yl) -2- (1-tert-butoxycarbany 1-1-methylethoxyimino) - acetamido]-3-chloromethyl-3-cepbem-4-carboxylate and 7— [3— (tert-butoxycarbonylamino) propionyl] amino-2 , 3- dihydro-lH-imidazo[l,2-b]pyrazole in the same manner as 30 in Example 1 as an amorphous solid. 1H-NMR(D2O) 1.51 (3H, s), 1.52 (3H, s), 2.33 (2H, t, J=6.4Hz), 3.26 (1H, d, J=17.9Hz), 3.53 (1H, d, J=17.9Hz), 3.31 (2H, t, J=6.4Hz), 4.15 (2H, t, J=8.7HZ), 4.33 (1H, q,J=8.7Hz), 4.42 (1H, q, J=8.7Hz), 4.95 (1H, d, 35 J=l.5Hz), 5.03 (1H, d, J=l5.lHz), 5.25 (1H, d, J=5.0Hz), 5.84 (1H, d, J=5.0Hz), 8 . 06 (1H, s) Preparation 11 tert-Butyl [2-(5-amino-l-methyl-lH-pyrazol-4- 38 WO 2004/039814 PCT/JP2003/013684 ylcatbamoyl) ethyl] carbonate The title compound was obtained from 4,5-diamino- 1-methylpyrazole sulfuric acid salt and N-[3-(tert- butoxycarbonylamino)propionyloxy]succinimide in the same 5 manner as in preparation 10. 1H-NMR(DMSO-d6) 1.3 8 (9H, s) , 2.35 (2H, t, J=7.1Hz), 3.l8 (2H, q, J=7.lHz), 3.50 (3H, s) , 4.90 (2H, s), 6.83 (1H, t, J=7.1Hz) , 7.14 (1H, s) , 9.06 (1H , s) AP-MS: m/z=283 10 Preparation 12 tert-Butyl (2-[1-methy1-5-(tritylamino)-1H- pyrazol-4—ylcarbamoyl] ethyl)carbamate The title compound was obtained from tert—butyl (2- (5-amino-l-methyl-lH-pyrazol-4-ylcarlbamoyl) thyl] - 15 carbamate in the same manner as in Preparation 6. 1H-NMR(DMSO-d6) 1.39 (9H, s) , 2. 08 (2h, t, J=7.1Hz), 2.71 (3H, s) , 3.04 (2H, q, J=7-lHz), 5.57 (1H, s) , 6.72, (1H, t, J=7.1Hz), 7.1-7.4 (16H, m) , 8.25 (1H, s) Example 4 20 7b[ (Z) -2- (5-Amino-l,2,4-thiadiazol-3-yl) -2-(l- carboxy-l-methylethoxyimino) acetamido] -3-[3-amino-4- (3- aminopropionamido) -methyl-l-pyrazolio]methyl—3-cephem- 4-carboxylate The title compound was obtained from benzhydryl 25 7 b [(Z) -2-(5-tert-butoxycarbonylamino-l,2,4-thiadiazol- 3-yl) -2- ( 1-tert-butoxycarbonyl-l-methylethoxyimino) - acetamidol-3-ch1oromethyl-3-cephem-4-carboxylate and tert-butyl (2-([l-methyl-5-(tritylamino)-lH-pyrazol-4- yLcarbamoyl]-ethyl] carbamate in the, same manner as in 30 Example 1. 1H-NMR(D2O) 1.53 (6H, s) , 2.89 (2H, t, J=6.5Hz), 3.20 and 3,47 (2H, ABq, J=18Hz), 3.34 (2H, t, J=6.5HZ), 3.75 (3H, 5), 4.99 and 5.21 (2H, ABq, J=16Hz), 5.25 (1H, d J=4.8HZ), 5.85 (1H, d, J=4.8Hz), 8.02 (1H, s) 35 ESI-MS: m/z=674 (m+Na) Preparation 13 To a solution of 1, 3-bis (tert-butoxycarbornyl) -2- (trifluoromethylsulfonyl)guanidine (22.3 g) in 39 WO 2004/039814 PCT/JP2003/013684 dichloromethane [100 ml] were added 4 , 5-diamino-l- methylpyrazole sulfuric acid salt (10 g) and triethylamine (33.2 ml) at 4°C, and the mixture was stirred at room temperature overnight. The reaction 5 mixture was poured into a mixture of ethyl acetate and water. The aqueous layer was separated, and the organic layer was washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated in vacuo. The concentrate was poured into 10 acetonitrile, and the resulting precipitate was collected by filtration and dried in vacuo to give 5- amino-4-[2' ,3'-bis(tert-butoxycarbonyl)quapidino1-1- methylpyrazole [11.62 g). 1H-NHR(DMSO-d6) 1.37 (9H, s), 1.50 (9H, s) , 3.52 (3H, 15 s) , 5.14 (2H, s) , 7.11 (1H, s) , 9.14 (IH, s), 3.1.5 (1H, s) ESl-MS: m/z=353 (M-H) Preparation 14 4- [2 ', 3 '-Bis (tert-butoxycarbonyl) quanidino] -1- 20 methyl-5- (tritylamino) pyrazole The title compound was obtained from 5-amino-4- [2 ' , 3 ' -bis (tert-butoxycarbonyl) guanidino] -1- methylpyrazole in the same manner as in Preparation 6, 1H-NMR(DMSO-d6) 1.37 (9H ,s) 1.50 (9H, s), 2.35 (3H, 25 s) , 5.88 (1H, s) , 7.17 (1H , s) . 7.21 (15H, m) , 8.85 (1H, a), 11.2 (1H, s) ESI-MS: m/z=597(M+H) Example 5 7 b- [(z) -2- [5-Amino-l, 2 , 4-thiadiazol-3-yl) -2- (1- 30 carboxy-1-methyletboxyimino)acetamido]-3-(3-amino-4- quanidino-2 -methy1-1 -pyrazo1io) methy 1- 3- cephem-4 - carboxylate The title compound was obtained from benzhydryl 7 b -[ (Z) -2- (5-amino-l,2,4-thiadiazol-3-yl) -2-(l-tert- 35 butoxycarbonyl-l-methylethoxyimino)acetamido]-3- iodomethyl-3-cephem-4-carboxylate and 4-[2 ' , 3'-bis ( tert- butoxycarbonyl) quanidino]-l-methyl-5- (tritylamino) pynazole in the same manner as in Example 1. 40 WO 2004/039814 PCT/JP2003/013684 1H-NMR (DMSO-d6) 1.53 (6H, s) , 3.25 and 3.57 (2H, ABq, J=l8Hz) , 3.75 (3H, S),0 5.00 and 5.18 (2H, ABq, J=l5Hz) , 5.21 (1H, d, J=4.9HZ) , 5.85 (1H, d, J=4.9Hz), 8.05 (1H, 5 Preparation 15 To a suspension of 4 , 5-diamino-l-methylpyrazole sulfuric acid 5 salt (305 g) in tetrahydrofuran (3.05 L) was added tert-butyl 2-[(2,5-dioxo-l-pyrrolidinyl)oxy]- 2-oxoethylcarbamate (415 g) under ice-cooling. To the 10 mixture was added diisopropylethylamine (556 ml) dropwise at a temperature below 10°C. The mixture was stirred at room temperature overnight. To the resulting solution were added brine and saturated aqueous sodium hydrogen carbonate solution, and the mixture was 15 extracted with ethyl acetate (3.0 L) . The aqueous layer was extracted with tetrahydrofuran/ethyl acetate=l/l (3.0 L) twice. The organic layer was dried over anhydrous macflie^ium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl 20 ether (1.0 L) and dried in vacuo to give tert-butyl 2- [ (5-amino-1 -methyl-1H-pyrazol-4-yl) amino ] -2- oxoethylcarbamate (307 g) . IR(Br) 3440, 3349, 1670, 1631, 1525, 1276, 1163, 1074, 1014, 860, 791 cm-1 25 1H-NMR(DHSO-d6) 1.39 (9H, s) , 3.44 (3H, s), 3.64 (2H, d, J=5.9Hz), 4.91 (2H, brs), 6.97 (1H, t, J=5.9Hz), 7.15 (1H, s) , 9.09 (1H, brs) Preparation 16 To a solution of tert-butyl 2- [ (5-amino-l-methyl- 30 lH-pyrazol-4-yl]amino] -2-oxoethylecarbamate (307 g) in N,N'-dimethylformamide (1.5 L) was added triphenylmethyl chloride (334 g) , To the mixture was added triethylamine (318 ml) dropwise. The mixture was stirred at room temperature for 1 hour. The reaction 35 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 41 WO 2004/039814 PCT/JP2003/013684 concentrated in vacuo. The residue was triturated with acetonitrile (1.5 L) and dried in vacuo to give tert- butyl 2-( l-methyl-5-(tritylamino)-lH-pyrazol—4- yl]amino]-2-oxoethylcarbamate (468 g) . 5 IR(KBr) 3336, 3280, 1724, 1683, 1599, 1234, 939, 704 cm-1 1H-NMR(DMSO-d6) 1.38 (9H, s) , 2.73 (3H, s), 3.38 (2H, d, J=5.8Hz), 5.58 (1H, s) , 6.94 (1H, t, J=5.8Hz), 7.11-7.35 (15H, m) , 7.21 (1H, s), 8.31 (1H, s) 10 ESI-MS: m/z=5l2.3(M+H+) Example 6 To a solution of benzhydryl 7—t(Z)-2-(5-amino- 1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1- metylethoxyimino) acetamido] — 3-chloromethyl-3—cephem—4- 15 carboxylate (489 g) in N,N-dimethylformamide (1.4 L) was added sodium iodide (102 g) . after stirring at room temperature for 1 hour, tert-butyl 2-{[l-methyl-5- (tritylamino) —lH-pyrazol-4-yl] amino)-2-oxoethylcarbamate (383 g) was added to the mixture. Stirring was 20 continued at 37oC for 24 hours. The resulting mixture was poured into water and extracted with, ethyl acetate. The organic layer was washed successively with water, 10% acqueous sodium thiosulfate solution, brine and 10% aqueous sodium trifluoroacetate solution, dried over 25 magnesium sulfate, filtered and evaporated in vacuo. The residue was dissolved in ethyl acetate (3.5 L), and the solution was dropwise added to diisopropyl ether (36 L) . The precipitate was collected by filtration. The filter cake was washed with diisopropyl ether and dried 30 in vacuo. The obtained solid (700 g) was dissolved in dichloromethane (1.4 L), and to the solution were added anisole (700 ml) and trifluoroacetic acid (2.1 L) successively. After stirring at room temperature for 4 35 hours, the reaction mixture was poured into diisopropyl ether (3 0 L) . The precipitate was collected by filtration. The obtained solid was washed with diisopropyl ether and dried in vacuo. The crude product 42 WO 2004/039814 PCT/JP2003/013684 was dissolved in water (3.5 L), and the pH of the solution was adjusted to 7.0 with 28% aqueous ammonia solution. The insoluble material was filtered off, and the pH of the filtrate was adjusted to 1 with 5 concentrated hydrochloric acid. The insoluble material was filtered off again. The filtrate was chromatographed on Diaion® HP- 20 eluting with 20% aqueous 2—propanol. The eluate was concentrated to about 3.0 L in vacuo, and 2.0M sulfuric acid (102 ml) 10 was added, to the concentrate. The mixture mas lyophilized to give the crude product. The crude product was purified by preparative HPLC (pH 7.0 phosphate buffer and acetonitrile), and the eluate containing a desired product was concentrated to 15 about 6 L in vacuo. The concentrate was adjusted to about pH 1 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 eluting with 20% aqueous 2-propanol. The eluate was concentrated to about 550 ml in vacuo, and 2.0M sulfuric acid (54.5 ml) 20 was added to the concentrate. To the mixture was added dropwise acetonitrile (880 ml), and the mixture was stirred at room temperature overnight. To the mixture was added acetonitrile (200 ml), and the mixture was stirred at room temperature for 2 hours. The resulting 25 white crystals were collected by filtration, washed with 25% aqueous acetonitrile and dried under reduced pressure to give 7[(Z)-2-(5-amino-l,2,4-thiadiazol-3— yl) —2— (l-carboxy-l-methylethoxyimino) acetamido] -3- [3- amino-4- (aminoacetyl)amino-2-methyl-l-pyrazoliolmethyl- 30 3-cephem-4-carboxylic acid hydrogen suIfate (72.5 g), IR(KBr) 1778, 1700, 1653, 1525, 1149, 1111, 617 cm-1 1H-NMR(D20] 1.61 (6H, s) , 3.22 and 3.45 (2H, ABq, J=l7.8Hz), 3.73 (3H, s), 4.03 (2H, s), 5.05 and 5.27 (2H, ABq, J=l5.8Hz), 5.25 (1H, d, J=4.8Hz), 5.87 (1H, d, 35 J=4.8az), 8.09 (1H, s) ESI-MS: m/z=638,2 t(M+H+) Example 7 A solution of 7-[(Z)-2-(5-amino-l,2,4-thiadizol- 43 WO 2004/039814 PCT/JP2003/013684 3-yl) -2- (1-carboxy-l-methylethoxyimino) acetamidol -3-(3- amino-4- [3- (2-aminoethyl) ureido] -2-methy1-1- pyrazolio)methyl—3-cephem-4-carboxylate (36 g) in water was purified by preparative HPLC utilizing ODS column. 5 The eluate containing a desired product was concentrated to about 1.5 L in vacuo. The concentrate was adjusted to about pH 1 with concentrated hydrochloric acid and chromatographed on Diaion® HP—20 (6 L) eluting with 20% aqueous 2-propanol. The eluate was concentrated to 10 about 800 ml in vacuo, and 2M sulfuric acid (17 ml) was added. The resulting solution was lyophilized to give a sulfuric acid salt as an amorphous powder (23.6 g). The powder was dissolved in water (71 ml) and ethanol (57 ml). After addition of seed crystals (310 15 mg) , which resulted in the precipitation of white solid, the mixture was stirred for 1 hour. A mixture of ethanol (47 ml) and water (37 ml) was added over 30 minutes, and ethanol (33 ml) was added over 2 0 minutes. Then the slurry was stirred for an additional 1.5 hour. 20 The precipitate was collected by filtration, washed with ethanol/water (60 ml/20 ml) and ethanol (60 ml) and dried to give 7-[(Z)-2- (5-amino-l,2,4-thiadiazol-3-yl)- 2- (1—carboxy-1—metnylethoxyimino) acetamido] -3 —[3—amino- 4— [3- (2—aminoethyl) ureido ]-2—methy1-1—pyrazolio] methyl- 25 3-cephem-4—carboxylic acid hydrogen sulfate as crystals (17.3 g). IR(KBr) 3353, 3183, 1778 , 1652, 1558 , 1403, 1321, 1143, 1118, 997, 619 cm-1 1H-NMR(D2O) 1.61 (6H, s), 3.10-3.55 (6H, m) , 3.71 (3H, 30 s), 5.02 and 5.23 (2H, ABq, J=16.7Hz), 5.25 (1H, d, J=4.9Hz), 5.87 (1H, d, J=4.9Hz), 7.91 (1H, s) ESI-MS: m/z=667 (M+H+) X-ray powder diffraction analysis (by Eigatu X-ray Diffraction system MultiFlex) 35 20 intensity 8.0 1286 12.7 586 13.8 423 44 WO 2004/039814 PCT/JP2003/013684 16.1 618 18.9 520 20.4 748 21.5 667 5 22.4 1058 23.3 944 24.0 618 25.5 813 26.7 472 10 279 537 28.5 455 31.3 390 X-ray: Cu/40 kV/30 mA Preparation 17 15 5-Amino-1- ethy1-4- nitrosopyrazole The title compound was obtained from 5-amino-l- ethylpyrazole in the same manner as in Preparation 3, 1H-NMR(DMSO-d6) d 1.21 (3H, t, J=7,lHz), 3.93 (2H, q, J=7.1Hz), 7.04 and 8. 53 (1H, s) , 8.10 and 8.15 (1H, brs) 20 APCI-MS: m/z=l41(M+H)+ Preparation 18 4,5-Diamino-l-ethylpyrazole sulfuric acid salt The title compound was obtained from 5-amino-l- ethyl-4-nitrosopyrasole in the same manner as in 25 Preparation 4. 1H-NMR(D2O) 1.36 (3H, t, J=-7.3Hz) , 4.10 (2H, q, J=7.3Hz), 7.77 (1H, s) ESI-MS: m/z=27 (M+H) + Preparation 19 30 5-Amino-4- [3- (tert-butoxycarbonylamino) - propionylamanol -l-ethylpyrazole The title compound was obtained from 4,5-diamino- 1—ethylpyrazole sulfuric acid salt in the same mannner as in Preparation 15. 35 1H-NMR(DMSO-d6) 1.24 (3H, t, J=7.2Hz), 1.37 (9H, s), 2.35 (2H, t, J=7.1HZ), 3.18 (2H, dt, J=7.1, 7.lHz), 3.35 (q, J=7.2Hz), 4.88 (2H, brs), 6.75-6.90 (1H, m), 7.17 (1H, s). 9.05 (1H, brs) 45 WO 2004/039814 PCT/JP2003/013684 APCI-MS: m/z=298(M+H)+ Preparation 20 4- [3- (tert-Butoxycarbonylamino)propionylamino] -1- ethyl-5—tripheaylmethylaminopyrazole 5 The title compound was obtained from 5-amino-4- [3- (tert-butoxycarbonylamino) propionylamino] —1- ethylpyrazole in the same manner as in Preparation 16. lH-NMR(DMSO-d6) 0.88 (3R, t, J=7.2Hz), 1.39 (9H, s) , 2.02 (2H ,t, J=7.1Hz), 2.95-3.20 (4H, m), 5.59 )(1H, brs), 10 6.60-6.75 (1H, m), 7.10-7.35 (16H, m), 8.04 (1H, brs) ESI-MS: m/z=540 (M+H)+, 562(M+Na)+ Example 8 7b[(Z)-2-(5-Amino-l,2,4-thiadiazol-3-yl)-2- (1- carboxy—1-methylethoxyimino) acetamido ) -3-[3-amino-4- (3- 15 aminopropionylamino) -2-ethyl-l-pyrazolio]me-tliyl-3 - cephem-4-carboxylate The title compound was obtained from benzhydryl 7 b [ (Z)-2- [5-amino-l,2,4-thiadiazol-3-yl)-2- (l-tert- butoxycarbonyl-1-methylethoxyimino) acetamido] -3- 20 iodomethyl—3-cephem-4-carboxylate and 4-[3-(tert- butoxycarbonylamino) propionylamino] -l-ethyl-5- triphenylmethylaminopyrazole in the same manner as in Example 1. IR(KBr) 3415, 1763, 1658, 1598, 1529, 1402, 1361 cm-1 25 1H-NMR(D2O) 1.33 (3H, t, J=7.2Hz), 1.53 (6H, s), 2.89 (2H, t, J=6.5Hz), 3.17 and 3.49 (2H, ABq, J=17.7Hz), 3.34 (2H, t, J=6.5HZ), 4,26 (2H, q, J=7.3Hz), 5.05 and 5.16 (2H, ABq, J=15.4H2), 5.26 (lH, dr J=4.8Hz), 5.85 (1H, d, J=4.8Hz), 8.03 (1H, s) 30 Preparation 21 tert-Butyl 2-[ (5-amino-l-ethyl-lH-pyrazol-4- yl) amino] -2-oxoethylcarbamate The title compound was obtained from 4,5-diamino- 1-ethylpyrazole sulftiric acid salt in the same manner as 35 in Preparation 15. 1H-NMR(DHSO-d6) 1.21 (3H, t, J=7.2Hz), 1.39 (9H, s), 3.64 (2H, d, J=6.0Hz), 3.86 (2H, d, J=7.2Hz), 4.88 (2H, brs), 6.90-7.00 (1H, m) , 7.17 (1H, s), 9.06 (1H, brs) 46 WO 2004/039814 PCT/JP2003/013684 ESI-MS: m/z=284 (M+H) + , 306(M+Na) + Preparation_2 2 tert-Butyl 2- {[l-eth.yl-5- (tritylamino) -1H-pyrazol- 4-yl] amino)-2-oxoethylcarbamate 5 The title compound was obtained from tert-butyl 2- [ (5-amino-1-ethyl-1H-pyrazol-4-yl) amino] -2- oxoethylcarbamate in the same manner as in Preparation 16. 1H-NMR(DMSO-d6) b 0.88 (3H, t, J=7.2Hz), 1.38 (9H, s), 10 3.16 (2H, q, J=7.2Hz), 3.31 (2H, d) , 5.59 (1H, brs) , 6.80-6.95 (1H, m), 7.10-7.40 (l6H, m), 8.03 (1H, brs) ESI-MS: m/z=526 (M+H)+, 548(M+Na}+ Example 9 7 b [ (Z) -2- (5Amino-1,2, 4-thiadiazol-3-yl) -2- (1- 15 carboxy-1 -methylethoxyimino) acetamido ] -3- [ 3 -amino-4- (aminoacetyl) amino-2-ethyl-l-pyrazolio] methyl-3-cephem- 4-carboxylate The title compound was obtained from bezhydryl 7 b [ (Z) -2-(5-amino-l, 2 , 4-thiadiazol-3-yl) -2- (l-tert- 20 butoxycarbony 1-1 -methylethoxyimino) acetamido] -3- iodomethyl -3-cephem-4-carboxylate and tert-butyl 2- [( 1- ethyl-5-(tritylamino) -lH-pyrazol-4-yl] amino } -2- oxoethylcarbamate in the same manner as in Example 1 - IR(KBR) 3444, 1761, 1635, 1626, 1445, 1406 cm-1 25 1H-NMR(D20) 1 . 33 [3H, t, J=7. 2Hz) , 1.53 (6H, s), 2.89 (2H, t, J=6.5Hz), 3.17 and 3.49 (2H, ABq, J=17.7Hz), 4.00 (2H, s), 4.28 [2H, q, J=7.2Hz), 5.06 and 5.17 (2H, ABq, J=15.4Hz) , 5.27 (1H, d, J=4.3Hz), 5.85 (1H, d, J=4. 8Hz) , 8.07 (lH, s) 30 Preparation 23 5-Amino-4-[2' ,3'-bis (tert-butoxycarbonyl)- quanidinol -l-ethylpyrazole The title compound was obtained from 1,3-bis(tert- butoxycarbonyl)-2- (trifluoromethylsulfonyl) quanidine and 35 4,5-diamino-l-ethylpyrazole sulfuric acid salt in the same manner as in Preparation 13. 1H-NMR(DMSO-d6) 1.22 (3H, t, J=7.1Hz), 1.37 (9H, s), 1.50 (9H, s) , 3.88 (2H, d, J=7.1Hz),, 5.12 (2H, brs) , 47 WO 2004/039814 PCT/JP2003/013684 7.14 (1H, s) , 9.16 (1H, brs) , 11.51 (1H, brs) ESI-MS: m/z=369(M+H)+ Preparation 24 4-[2' ,3 '-Bis(tert-butoxycarbonyl)guanidino]-l- 5 ethyl-5-triphenylmethylaminopyrazole The title compound was obtained from 5-amino- 4- [2',3'-bis(tert-butoxycarbonyl)quanidino]-1- ethylpyrazole in the same manner as in Preparation 16. 1H-NMR(DMSO-d6) 0.8 6 (3H, t, J=7.1Hz), 1.3 8 (9H, s), 10 1.49 (9H,s) , 5. 85 (1H, brs) , 7.10-7.30 (16H, m) , 8.80 (1H, brs), 11.14 (lHr, brs) ESI-MS: m/z = 611 (M+H) +, 633 (M+Na)+ Example 10 7 b -[(Z) -2-(5-Amino-l,2,4-thiadiazol-3-yl)-2- (1- 15 carboxy-1 -methylethoxyimino) acetamidol-3-[3-amino-2- ethyl-4-quanidino-l-pyra zolio]methyl-3-cephem-4- carboxylate The title compound was obtained from benzhydryl 7 b -Z) -2- (5-amino-1,2,4 -thiadiazol -3-yl) -2 - (1 -tert- 20 butoxycarbonyl-l-methylethoxyimino) acetamido] -3- iodomethyl-3-cephem-4-carbonylate and 4- [2 ' , 3 ' -bis ( tert- butoxycarbonyl) quanidino] -l-ethyl-5- triphenylmethylaminopyrazola in the same manner as in Example 1. 25 IR(KBr) 3437, 1760, 1658, 1625, 1406, 1065 cm-1 lH-NMR(D20) 1.35 (3H, t, J=7.3Hz), 1.53 (6H, a), 3.26 and 3.61 (2H, ABq, J=17.8Hz), 4.25 (2H, q , J=7.3Hz), 5.06 and 5.17 (2H, ABq, J=15.7Hz), 5.29 (1H, d, J=4 . 8Hz) , 5.85 (1H, d, J=4.8Hz), 8.06 (lH, s) 30 Example II To a suspension of benzhydryl 7b-[ (Z)-2-{5-amino- 1, 2 ,4-thiadiazol-3-yl) -2- (l-tert-butoxycarbonyl-l- methylethoxyimino) acetamido] -3-iodomethyl-3-cephem-4- carboxylate (5 00 g) in N,N—dimethylformamide (2,5 L) was 35 added 4-[2',3'-bis(tert-butoxycarbonyl)quanidino]-l- methyl-5-triphenylmethylaminopyrazole (419 g) and the mixture was stirred at room temperature for 16 hours. The reaction mixture was added to a mixture of ethyl 48 WO 2004/039814 PCT/JP2003/013684 acetate and water. The organic layer was washed with water, brine and 10% aqueous sodium trifluoroacetat solution and then dried over magnesium sulfate. The magnesium sulfate was filtered off, and the filtrate was 5 evaporated to 3.3 kg under reduced pressure. The concentrate was poured into diisopropyl ether (33 L), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene 10 chloride (1500 ml) were added anisole (500 ml) and trifluoroacetic acid (1500 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. The crude 15 product was dissolved in water (3.5 L), and the pH of the solution was adjusted to 7.0 with 28% aqueous ammonia solution. The insoluble material was filtared off, and the pH of the filtrate was adjusted to 1 with concentrated hydrochloric acid. The insoluble material 20 was filtered off, again. The filtrate was chromatographed on Diaion® HP-20 eluding with 20% aqueous 2-propanol, The eluate was concentrated to about 3.0 L in vacuo. To the concentrate was added 2.0M sulfuric acid (150 ml), and the mixture was lyophilized 25 to give the crude product. The crude product was purified with preparative HPLC utilising ODS column (pH 7.0 phosphate buffer and acetonitrile). The eluate containing a desired product was concentrated to about 6 1 in vacuo. The concentrate was adjusted to about pH 1 30 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 eluting with 20% aqueous 2-propanol. The eluate was concentrated to about 1.5 L in vacuo. To the concentrate was added 2.0M sulfuric acid (SO ml), and the mixture was lyophilized to give 7 b - [(Z) -2- (5- 35 amino-1,2,4-thiadiazol-3-yl)-2- (l-carboxy-1- methylethoxyimino) acetamido] -3-(3-amino-4—gnanidino-2- methyl-1-pyrazolio) methyl—3-cephem—4-carboxylic acid hydrogen sulfate (48.5g). 49 WO 2O041&398I4 PCT/JP30O3/013684 IR(KBr) 1776, 1714, 1577, 1651, 1402, 1112 cm-1 1H-NMR(D2O) 1.61 (6H, s) , 3.28 and 3,58 (2H, ABq, J=l7.8Hz), 3.74 (3H, s), 5.15 and 5.23 (2H, ABq, J=15.7Hz), 5.27 (1H, d, J=4.8Hz), 5.88 (1H, d, J=4.8Hz), 5 8.07 (1H, s) ESI-MS: m/z=623.2(M+H+) Preparation 25 To a suspension of 4 ,5-diamino-l(2- hydroxyethyl)pyrazole sulfuric acid salt (2.4 g) in 10 methylene chloride (40 ml) were added N- ethyldisopropylamine (2.1 ml) and N- [3- (tert- butoxycarbonyXamino)propionyloxy] succinimide (2.3 g) under ice-cooling, and the mixture was stirred at room temperature for 6 hours. To the reaction mixture were 15 added brine (40 ml) and saturated aqueous sodium hydrogen carbonate solution (20 ml), and the mixture was extracted with a mixture of ethyl acetate and 2-propanol (3:1, 60 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in 20 vacuo. The residue was triturated with diethyl ether to give 5-amino-4- [3- (tert-butoxycarbonylamino) propionyl] - amino-1-(2-hydroxyethyl)pyrazole (l.65 g) as a solid. 1H-NMR(DMSO-d6) 1.38 (9H, s) , 2.35 (2H, t, J=7.3Hz), 3.16-3.20 (2H, m), 3.62-3.65 (2H, m), 3.90 (2H, t, 25 J=6.0HZ), 4.85 (2H, brs) , 4.92 (1H, t, J=5.0HZ), 6.84 (1H, t, J=5.5Hz), 7.20 (1H, s), 9.09 (1H, brs) Example 12 7 b - [ (Z) -2- (5-Amino-l ,2 , 4-thiadiazol-3-yl) -2- (1- carboxy-l-methylethoxyimino)acetamido]-3-[3-amino-4-(3- 30 aminopropionamido)-2- (2-hydroxyethyl)-1- pyrazoliolmethy1-3-cephem-4-carboxylate The title compound was obtained from benzhydryl 7 b [ (Z) -2- (5-tert-butoxycarbonylamino-1, 2 , 4-thiadiazol- 3-yl)-1- (1-tert-butoxycarbonyl-l-methylethoxyimino)- 35 acetamido]-3-chloromethyl-3-cephem-4-carboxylate and 5- amino-4-[3- (tert-butoxycarbonylamino)propionyl] amino-1- [2-hydroxyethyl)pyrazole in the same manner as in Example 1 as an amorphous solid. 50 WO 2004/039814 PCT/JP2003/013684 1-H-NMR(D2O) 1.51 (6H, s) , 2.88 (2H, t, J=6 . 4Hz) , 3.15 (1H, d, J=17.9Hz), 3.48 (1H, d, J=17.9Hz), 3.32 [2H, t, J=6,4Hz), 3.88 (2H, t, J=4.3Hz), 4.39 (lH, dt, J=l6.5Hz, 4.8Hz), 4.42 (1H, dt, J=16.5, 4.8Hz), 5.06 (1H. d, 5 J=15.lHz), 5.11 (1H, d, J=15.lHz), 5.25 (lH, d, J=5.0Hz), 5.83 (lH, d, J=5.0Hz), 8.05 (1H, s) Preparation 26 To a solution of 4-formyl-4,5,6,7- tetrahydropyrazolo(1 .5-alpyrimidine (1.51 g) in sulfuric 10 acid (7.5 ml) was added potassium nitrate (111 g) under ice-cooling. The mixture was stirred at room temperature for 17 hours. The reaction mixture was added to ice (100 g) . The crystalline residue was collected by filtration and dried in vacuo to give 3- 15 nitro-4,5 ,6,7-tetralhydropyrazolo [1,5-alpyrimidine (0.63 g) as a solid. 1H-NMR(DMSO-d6) 2.00-2.05 (2H, m) , 3.30-3.36 (2H, m), 3.99 (2H, t, J=6.0Hz), 7.85 (1H, s), 7.89 (1H, s) Preparatjon27 20 A solution of 3-nitro-4,5,6,7- tetrahydropyrzolo [1,5-alpyrimidine (1.68 g) in a mixture of sulfuric acid (0.6 ml) , acetic acid (100 ml) and water (10 ml) was treated with 10% palladium on carbon (0.5 g) under a hydrogen atmosphere at room 25 temperature for 6 days. After the catalyst was filtered off, the filtrate was concentrated in vacao. The residue was triturated with ethanol and dried in vacuo to give 3-amino-4 ,5, 6 , 7-tetrahydropyrazolo [1 , 5- alpyrimidine sulfuric acid salt (2.3 g) as a solid. 30 1H-NMR (DMSO-d6) 1.97-2.01 (2H, m), 3.22 (2H, t, J=5.0Hz), 3.98 (2H, t, J=6.0Hz), 7.22 (1H, s) Preparation 28 To a solution of 3-amino-4,5,6,7- tetrahydropyrazolo[l,5-alpyrimidine sulfuric acid salt 35 (2.96 g) and N-ethyldiisopropylamine (3.88g) in methylene chloride (70 ml) was added 1,3-bis(tert- butoxycarbonyl)-2-(trifluoromethanesulfonyl) guanidine (3.91 g) . The mixture was stirred at room temperature 51 WO 2004/039814 PCT/JP2003/013684 for 150 minutes. The reaction mixture was washed with saturated aqueous sodium hydrogen, carbonate solution. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The 5 residue was purified by column chromatography on silica gel eluting with 2% methanol/chloroform to give 3-[2,3- bis (tert-butoxycarbonyl) guanidino ]-4,5 , 6 , 7-, tetrahydropyrazolo[l,5-alpyrimidine (3 . 4 g) as a solid. lH-NMR(CDCl3) 1.48 (9H, s), 1.52 (9H, s), 2.12-2.14 (2H, 10 m) , 3.33-3.37 (2H, m) , 4.08 (2H, t, J=6.0Hz), 6.17 (lH, brs), 7.16 (1H, s) , 9.87 (1H, brs), 11.39 (1H, brs) Example 13 To a solution of benzhydryl 7-[(Z)—2-[5—tert- butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2-fl-tert- 15 butoxycarbonyl-1-methylethoxyimino) acetamido] -3- chloromethyl-3-cephem-4-carboxylate (1.0 g) in ,N,N- dimethylformamide (2.0 ml) was added sodium iodide (181 mg) , and the mixture was stirred at room temperature for 30minutes. To the reaction mixture were added 3-[2,3- 20 bis(tert—butoxycarbonyl) guanidino]-4,5,6 ,7- tetrahydropyrazolo[1,5-a]pyrimidine (571 mg) and methylene chloride (2.0 ml). The whole mixture was stirred at room temperature for 7 hours. To the reaction mixture were added ethyl acetate (100 ml) and 25 water (50 ml) . The aqueous layer was separated, and the organic layer was washed with 10% aqueous sodium trifluoroacetate solution and brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into 30 diisopropyl ether (150 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 35 trifluoroacetic acid (2.0 ml), and the mixture was stirred at room temperature for 4 hours. The reaction mixture was poured into diisopropyl ether (150 ml), and the resulting precipitate was collected by filtration 52 WO 2004/039814 PCT/JP2003/013684 and driad in vacuo to give a crude product (570 mg) , 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 5 was adjusted to about pH 3 with concentrated hydrochloric acid and chromatographed on Diaionâ HP-20 [Mitsubishi Chemical Corporation] eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo and lyophlized to give 7-[(Z)-2- 10 (5-amino-1,2,4—thiadiaizol-3-yl) -2— (l-carboxy—1- methylethoxyimino) acetamido] -3- [3-guanidino-4,5,6,7- tetrahydro -l-pyrazolo [1 , 5-alpyrimidinio] methy 1- 3 - cephem- 4-carboylate (51mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s) , 1.53 (3H, s), 2.05-2.25 (2H, 15 m), 3.26 (1H, d, J=l7.4Hz), 3. 56 (lH, d, J=l7.4Hz], 3.30-3.45 (2H, m), 4.15 (2H, t, J=6.0Hz), 4.93 (1H, d, J=15.6Hz), 5.15 (1H, d, J=15.6Hz), 5.25 (1H, d, J=4.8Hz), 5.84 (1H, d, J=4.8Hz), 7.99 (lH, s) Preparation 29 20 To a solution of 7-amino-2 ,3-dihydro-lH- imidazol[1,2-b]pyrazole sulfuric acid salt (4.4 g) , 4- (dimethylamino) pyridine (244 mg) and triethylamine (8.10 g) in chloroform (45 ml) was added 1,3-bis(tert- butoxycarbonyl) -2- (trifluromethanesulfonyl) guanidine 25 (10.18 g), The mixture was stirred at room temperature for 2 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 30 magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl ether to give 7- [2 , 3-bis (tert-butoxycarbonyl) guanidino] -2 , 3- dihydro-lH-imidazo [1,2,-b]pyrazole (4.6 g) as a solid. 1H-NMR(CDCl3) 1.49 (9H, s), 1.52 (9H, s) , 3.57-4.01 (2H, 35 m), 4.21 (2H, t, J=7.8Hz), 5.30 (1H, brs), 7.19 (1H, s), 9.86 (1H, brs), 11.32 (lH, brs) Example 14 7 b [(Z) -2-(5-Aminno-l,2,4-thiadiazol-3-yl)-2- (1- 53 WO 2004/039814 PCT/JP2003/013684 carboxy-l-methylethoxyimino) acetamido]-3- [7-guanidino- 2 ,3-dihydro-5- (lH-imidazo [1,2-b] pyrazolio) ]methyl-3- cephem- 4- carboxy1at e The title compound was obtained from benzhydryl 5 7 b - [ (Z) -2- (5-tert-butoxycarbonylamino-l ,2 ,4-thiadiazol- 3-yl) - 2-(l-tert-butoxycarbonyl-l-methylethoxyimino) - acetamido]-3-chloromethy1-3-cephem-4-carboxylate and 7 - [2 ,3-bis (tert-butoxycarbonyl) guanidino] -2, 3-diydro-lH- imidazo[1,2-b]pyrazole in the same manner as in Example 10 13 as an amorphous solid. 1H-NMR(D2O) 1.51 (3H, s), 1.52 (3H, s), 3.35 (1H, d, J=l7.9Hz), 3.61 (1H, d, J=17.9Hz), 4.19 (2H, t, J=8.7Hz), 4.37 (lH, q, J=8.7Hz), 4.47 (1H, q, J=8,7Hz), 5,00 (1H, d, J=15,lHz), 5.04 (1H, d, J=15.1HZ), 5. 26 (1H, d, 15 J=4.8Hz), 5.84 (1H, d, J=4.8HZ), 8.13 (1H, s) Praparation 30 To a salution of 5-amino-l— (2- hydroxyethyl)pyrazole (6.35 g) in a mixed solvent of ethanol (25 ml) and concentrated hydrochloric acid 20 [0.035 ml) was added dropwise isomyl nitrite (7.03 g) The mixture was stirred at room temperature for 17 hours. The crystalline residue was collected by filtration and dried in vacuo to give 5-amino-l-(2—hydroxyethyl)-4- nitrosopyrazole (4.0 g) as a solid. 25 1H-NMR(DMSO-d6) 3.68 (2H, t, J=5.5Hz), 3.94 (2H, t, J=5.5Hz], 4.89 (1H, br), 8.06(2H, br), 8.53 (1H, s) Preparation 31 A solution of 5-amino-l-(2-hydroxyethyl)-4- nitrosopyrazole (9 7 g) in a mixed solvent of sulfuric 30 acid (34 ml) and water (2000 ml) was treated with 10% paLLadium on carbon (10 g) under a hydtogen atmosphere at room temperature for 4 days, After the catalyst was filtered off, the filtrate was concentrated in vacuo. The residue was triturated with methanol and dried in 35 vacuo to give 4,5-diamino-l-(2-hydroxyethyl)pyrazole sulfuric acid salt (90.2 g) as a solid. lH-NMR(DMSO-d6) 3.66 (2H, t, J=5.5Hz), 3.9 5 (2H, t, J=5.5Hz), 7.25 (1H, s) 54 WO 2004/039814 PCT/JP2003/013684 Preparation_32 To a suspension of 4,5-diamino-l- (2- hydroxyethyllpyrazole sulfuric acid salt (50.0) g) in chloroform (500 ml) were added 4-(dimethyllamino) pyridine 5 (2.54 g) , triethylamine (116 ml) and 1,3-bis(tert- butoxycarbonyl)-2-(trifluoromethanesulfonyl) guanidine (106 g) • The mixture was stirred under reflux for 2 hours. After cooling on an ice bath, the reaction mixture was washed successively with water, 4% aqueous 10 nitric acid solution, water and aqueos sodium hydrogen carbanate solution. The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was triturated with a mixed solvent of ethyl acetate (50 ml) and diethyl ether (200 ml) to give 5- 15 amino-4- [ 2 , 3-bis( tert-butoxycarbonyl )guanidino]-l-(2- hydroxyethyl pyrazole (50 g) as a solid. 1H-NMR(CDCl3 ) 1.47 (9H, s) , 1.53 (9H,s) , 3.28 (1H, br) , 4.02-4.05 (4H, ml), 4.65 (2H, br) , 7,22 (1H, s) , 9,95 (1H, br), 11.55 (1H, br) 20 Example 15 7 [ (Z) -2-(5-Amino-l,2,4-thiadiazol-3-yl)-2- (1- carboxy-1- methylethoxyimino) acetamido]-3- [3-amino-4- guanidino-2- (2-hydroxyethyl) -l-pyrazolio]methyl-3- cephem- 4 - carboxy1ate 25 The title compound was obtained from benzhydryl 7 b - [ (Z) -2- (5-tert-butoxycarboxylamino-l, 2 , 4-thiadiazol- 3-yl) -2- (l-tert-butoxycarbonyl-l-methylethoxyimino) - acetamido]-3-chloromethyl-3-cephem-4-carboxylate and 5- amino-4- [2,3-bis (tert-butoxycarbonyl) guanidino] -1- (2- 30 hydroxyethyl)pyrazole in the same manner as in Example 13 as an amorphous solid. 1H-NMR(D2O) 1.52 (3H ,s), 3.21 (1H, d, J=l7.9Hz), 3.59 (1H, d, J=17.9Hz), 3.90 92H, t, J=4.8Hz), 4.35-4.50 (2H, m) , 5.07 (1H, d, J=14.9Hz), 5.11 (1H, d, J=14.9Hz), 5.28 35 (1H, d, J=5.0Hz), 5.84 (19, d, J=5.0Hz), 8.09 (1H, s) Preparation _3 3 To a solution of 7- [2,3-bis(tert- butoxycarbonyl) guanidino) -2 ,3-dihydro-lH-imidazo [1,2- 55 WO 2004/039814 PCT/JP2003/013684 b]pyrazole (1.83 g) in pyridine (10 ml) was added triphenylmethyl chloride (1.67 g) . The mixture was stirred at 50°C for 5 hours. After cooling, chloroform (50 ml) was added to the reaction mixture, and the 5 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 purified by 10 column chromatography on silica gel eluting with 2% methano1/chloroform to give 7- [2 ,3-bis (tert- butoxycarbonyl)guanidino]-l-triphenylmethyl-2,3-dihydr- lH-imidazo[1,2-blpyrazole (1.57 g) as a solid. 1H-NMR(CDC13) 1.47 (9H, s) , 1.48 (9H, s), 3.50 (2H, t, 15 J=7.8Hz), 3.92 (2H, t, J=7.8Hz), 7.07-7.26 (l0H, m), 7.53-7.54 (6H, m) , 8.34 (1H, brs), 11.12 (1H, brs) Example 16 To a solution of benzhydryl 7-[(Z)—2-(5-amino- 1,2,4-thiadiazol-3-yl) —2-(1-tert-butoxycarbonyl-l- 20 methylethoxyimin) acetamido]-3—iodomethyl—3-cephem—4— arboxylate (819 mg) in N,N-dimethylformamide (2.4 ml) as added N-(trimethylsilyl)acetamide (656 mg), and the ixture was stirred at room temperature for 30 minutes. To the reaction mixture was added 7-[2,3-bis(tert- 25 butoxycarbonyl)guanidino]-l-triphenylmethyl-2,3-dihydro- lH-imidazo[1,2-b]pyrazole (730 mg) . The whole mixture was stirred at room temperature for 6 hour a. To the resulting reaction mixture were added ethyl acetate (100 ml) and water (50 ml) . The aqueous layer was separated, 30 and the organic layer was washed with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into 35 diisopropyl ether (12 0 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene 56 WO 2004/039814 PCT/JP2003/013684 chloride (2.0 ml) were added anisole (O.67 ml) and trifluoroacetic acid (1.34 ml), and the mixture was stirred at room temperature for 4 hours. The reaction mixture was poured into diisopropyl ether (l20 ml). The 5 resulting precipitate was collected by filtration and dried in vacuo to give a crude product (430 mg) , 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 10 to about pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propsnol, The eXuate was concentrated to about 30 ml in vacuo and lyophilized to give 7-[(Z)-2-(5-amino-l,2,4-thiadiazol- 15 3-yl) -2-(l-carboxy-l-methylethoxyimino) acetamido]-3-[7- guanidino-2 ,3-dihydro-5- (lH-imidazo [1,2- b]pyrazolio)]methyl-3-cephem-4-carboxylate (20.4 mg) as an amorphous solid. IH-NMR(D2O) 1.51 (3H, s), 1.52 (3H, s), 3.35 (1H, d, 20 J=17.9Hz), 3.61 (1H, d, J=17.9Hz), 4,19 (2H, t, J=3.7Hz), 4.37 (1H, q, J=8.7Hz), 4.47 (1H, q, J=8.7Hz), 5.00 (1H, d, J=15.1Hz), 5.04 (1H, d, j=l5.1Hz), 5.26 (1H, d, J=4,8Hz), 5.84 (1H, d, J=4.8Hz), 8.13 (lH, s) Preparation 34 25 To a suspension of 1,1 '-carbonyldiimidazole (1.94 g) in methylene chloride (20 ml) was added tert-butyl N- (3-aminopropyl)carbamate (2.30 g), and the mixture was stirred at room temperature for 1 hour. To the reaction mixture were added N-ethyldiisopropylamine (2.56 g) and 30 4,5-diamino-l-methylpyrazole sulfuric acid salt (2.10 g) , and the mixture was stirred at 30oC for 3 days. The reaction mixture was concentrated in vacuo, The residue was purified by column chromatocgraphy on silica gel eluting with 6% methanol/chloroform to give 5-amino-4- 35 (3- (3-[(tert-butoxycarbonyl)amino]propyl]ureido)—1- methylpyrazole (1.75 g) as a solid. 1H-NMR(DMSO-d6) 1.37 (9H, s) , 1.43-1.49 (2H, m) , 2,89- 2.33 (2H, m) , 2.98-3.01 (2H, m) , 3.50 (3H, s) , 4.79 (2H, 57 WO 2004/039814 PCT/JP2003/013684 br) , 5.85 (H, br) 6.77 (1H, br), 6.96 (1H, s) , 7.12 (lH, br) Examaple 17 To a solution of benzhydryl 7-[(Z)-2-(5-tert- 5 butoxycarbonylamino-1-,2, 4-thiadiazol-3-yl) -2-(l-tert- butoxycarbonyl-l-methylethoxyimino) acetamido] -3- chloromethyl-3-cephem-4—carboxylate (1.0 g) in N,N- dimethylformamid (2.0 ml) was added sodium iodide (199 mg) , and the mixture was stirred at room temperature for 10 30 minutes. To the reaction mixture was added 5-aimino— 4— (3 — [3—[(tert-butoxycarbonyl)amino]propy1)ureido) - l- methylpyrazole (415 mg) and the whole mixture was stirred at 32°C for 24 hours. To the resulting reaction mixture were added ethyl acetate (50 ml) and water (50 15 ml) . The aqueous layer was separated, and the organic layer was washed with 10% aqueous sodium trifluoroacetate solution and brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate 20 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 (3.6 ml) were added anisole (1.2 ml) and trifluoroacetic acid (2.4 ml). The resulting 25 solution was stirred at room temperature for 4 hours and poured into diisopropyl ether (100 ml) . The resulting precipitate was collected by filtration and dried in vacuo to give a crude product (939 mg) , which was purified by preparative HPLC utilizing ODS column. The 30 eluate containing a desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to about pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propartol. The 35 eluate was concentrated to about 30 ml in vacuo and lyophilized to give 7-[ (Z) -2- (5-amino-l, 2 , 4-thiadiazol- 3-yl)—2—(1-carboxy-l-methylethoxyimino)acetamido]-3—(3— amino-4- [3- (3-aminopropyl) ureido] — 2-methyl-1- 53 WO 2004/039814 PCT/JP2003/013684 pyrazolio )methyl-3-cephem-4-carboxylate (53 mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s), 1.85-1.88 (2H, m) , 3.03 (2B, t, J=8Hz) , 3.22(2H, t, J=l8Hz), 3.26 (2H, 5 t, J=7Hz), 3.49 (1H, d, J=l8Hz), 3.72 (3H, s), 4.96 (1H, d, J=15Hz), 5.16 (1H, d, J=15Hz), 5.25 (1H, d, J=5Hz), Preparation 35 To a suspension of 1,1'-carbonyldiimidazole (973 10 mg) in methylene chloride (10 ml) was added tert-butyl N-(2-aminoethyl) carbamate (1.11 g) under ice-cooling, and the mixture was stirred at room temprature for 2 hours. To the reaction mixture were added N- ethyldiisopropylamnie (1.28 g) and 3-amino-4 ,5 , 6 , 7- 15 tetrahydropyrazolo[1,5-alpyrimidine sulfuric acid salt (1.18 g), and the mixture was stirred at 50oC for 6 hours. The reaction mixture was washed with brine. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo, The residue was 20 purified by column chromatography on silica gel eluting with 5% methanol/chloroform to give 3-(3-(2-[(tert- butoxycarbonyl)amino]ethyl)ureido)-4,5,6,7- tetrahydropyrazolo [1,5-alpyrimidine (150 mg) as a solid. 1H-NMR(CDCl3) 1.43 (9H, s) , 2.11-2.16 (2H, m) , 3.22- 25 3.35 (6H, m), 4.09 (2H, t, J=7Hz), 4.63 (lH, br), 5.14 (2H, br), 5.69 (1H, br), 7.17 (1H, s) Example 18 7 b -[(Z)-2-(5-Amino-l,2,4 thiadiazol-3-yl)-2-(l- carboxy-l-methylethoxyimino)acetamido]-3-(3-[3-(2- 30 aminoethyl ureidol-4 , 5 ,6 ,7-tetrahydro-1-pyrazol[1,5- al pyrimidinio ] methyl-3-cephem-4-carboxylate The title compound was obtained from benzhydryl 7 b -[(Z)-2-(5-tert-butoxycarbonylamino-l,2,4-thiadiazol- 3-yl)-2-(1-tert-butoxycarbonyl-l-methylethoxyimino)- 35 acetamidol-3-chloromethyl-3,-cephem-4-carboxylate and 3- (3-(2- [(tert-butoxycarbonyl)amino]ethyl]ureido)-4,5,6,7- tetrahydropyrazolo [1,5-alpyrimidine in the same manner as in Example 17 as an amorphous solid, 59 WO 2004/039814 PCT/JP2003/013684 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s), 2.09-2.21 (2H, m), 3.13 (2H, t, J=6Hz), 3.24 (1H, d, J=l8Hz), 3.35-3.52 (5H, m), 4,12-4.15 (2H, m), 4.88 (1H, d, J=16Hz), 5.13 (lH, d, J=16Hz) , 5.25 (1H, d, J=5Hz) , 5 . 85 (1H, d, 5 J=5Hz) , 7.8 3 (lH, s) Preparation 36 To a suspension of 1 ,1' -carboxyldiimidazole (97 3 mg) in methylene chloride (10 ml) was added 0-[2-(tert- butoxycarbonylamino) ethyl ]hydroxylamine (l.ll g) under 10 ice-cooling, and the mixtuire was stirred at room temperature for 2 hours. To the reaction mixture were added N-ethyldiisopropylamine (1.28 g) and 4,5-diamino- 1—methylpyrazole sulfuric acid salt (1.05 g) , and the mixture was stirred under reflux for 4 hours . The 15 reaction mixture was washed with 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 10% methanol/chloroform to give 5-amino-4—(3—(2- 20 [(tert-butoxycarbonyl)amino]ethoxy]ureido]-1- methylpyrazole (255 mg) as a solid. 1H-NMR(DMSO-d6) 1.38 (9H, s), 3.19-3.20 (2H, m) , 3.51 (3H, s), 3.72 (2H, t, J=6Hz) , 4.86 (2H, br), 6.95 (1H, br), 7.06 (1H, s), 8.02 (1H, brs), 9.15 (1H, brs) 25 Example 19 7 b [(Z)-2- (5-Amino-1,2, 4-thiadiazo1-3-yl) -2- 1 - carboxy- 1 -methylethoxyimino) acetamido ] -3 - (3—amino- 4 - [ 3 - (2-aminoethoxy)ureido]-2-methyl-l-pyrazolio)methyl-3- cephem-4-carboxylate 30 The title compound was obtained from benzhydryl 7 b -[(Z)—2-(5-tert-butoxycarbonylamino-1,2,4—thiadiazol- 3-yl) -2-(l-tert-butoxycarbonyl-l-methylethoxyimino) - acetamido]-3-chloromethyl-3-cephem-4-carboxylate and 5- amino-4-(3-[2-[(tert-butoxycarbonyl)amino] ethoxy)- 35 ureido)-1-methylpyrazole in the same manner as in Example 17 as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s) , 1.53 (3H, s), 3.21 (1H, d, J=18Hz), 3.33 (2H, t, J=5Hz), 3.47 (lH, d, J=18Hz), 3.74 60 WO 2004/039814 PCT/JP2003/013684 (3H, s), 4.17 (2H, t, J=5Hz), 4.59 (1H, d, J=l5Hz), 5.17 (1H, d, J=l5Hz), 5.26 (1H, d, J=5Hz), 5.8 6 (IH, d, J=5Hz), 7.93 (1H, s) Preparation 37 5 To a suspension of 1,1'-carbonyldiimidazole (1.95 g) in methylene chloride (20 ml) was added tart-butyl N- (2-aminoethyl)carbamate (1.92 g) under ice-cooling, and the mixture was stirred at room temperature for 2 hours. To the reaction mixture were added N- 10 ethyldiisopropylamine (2.59 g) and 7-amino-2,3-dihydro- lH-imidazo(1,2-b]pyrazole sulfucic acid salt (2.2.2 g) , and the mixture was stirred at room temperature for 16 hours. To the reaction mixture were added trityl chloride (9.0 g) and triethylamine (3.0 g) . The mixture 15 was stirred at room temperature for 24 hours. The reaction mixture was washed -with 10% aqxteous citric acid solution, brine and saturated aqueous sodium hydrogen carbonate solution, The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated 20 in Vacuo. The residue was purified by column chromatography on silica gel eluting with 3% methanol/chloroform to give 7-(3-(2-[(tert- butoxycarbonyl) aminolethyl]ureidol-2,3-dihydro-l- tritylimidazo [ 1, 2-b] pyrazole (8O0 mg) as a solid, 25 1H-NMR(CDCl3) 1.43 (9H, s) , 3.19 (4H, br) , 3.69 (1H, brs), 3.78-3.85 (4H, m), 4.51 [1H, br), 5.07 (lH, br), 7.20 (1H, s), 7.26-7.34 (9H, m), 7.45-7.47 (6H, m) Example 20 To a solution of benzhydryl 7- [(Z) -2- (5-amino- 30 1,2,4-thiadiazol-3-yl)-2-(l-tert-butoxycarbonyl-l-***** methyletho3iyimitio) acetamido] "3-iodorttethyl-3-cephenti"4" earboxylate (820 mg) in W,N-dimethylformamide (2^4 ml) was added W— (trimetliyl^ilyl) acetamMe (656 mg) , and the miHture was stirred at room temperature for 30 minutes, 35 To the reaction fliixture was added 7-t3-{2-[{tert- butoxycarbonyl)amino]ethyljureido)'2,3-dihydro-l- tritylimidaao[1,2-b]pyrasole [700 mg) , and the whole mixture was stirred at room temperature tor 6 hours. To 61 WO 2004/039814 PCT/JP2003/013684 the resulting reaction mixture were added ethyl acetate (50 ml) and water (50 ml), The aqueous layer was separated, and the organic layer was washed with 10% aqueous sodium trifluoroacetate solution and brine, 5 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 a solution of the 10 resulting solid in methylene chloride (3,0 ml) were added anisole (1.0 ml) and trifluotoacetic acid (2,0 ml). The resulting solution was stirred at room temperature for 4 hours and poured into diisopropyl ether (120 ml). The resulting precipitate was collected by filtration 15 and dried in vacuo to give a crude product, (830 mg) , 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 with concentrated. 20 hydrochloric acid and chromatographed on Diaion® 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 7 b -[(z)-2- (5-amino-l ,2 , 4-thiadiazol-3-yl) -2- (1-carboxy-l- 25 methylethoxyimino) acetamido] —3 —(7—(3— 2— aminoethyl)ureido] —2,3-dihydro—5-(1H-imidazo[1,2- b]pyrazolio) methyl-3-cephem-4-carboxylate (28.5 mg) as an amorphous solid. 1H-NMR(D2O) 1.53 (3H, s), 1.54 (3H, g), 3.14 (2H, t, 30 J=6Hz), 3.29 (1H, d, J=18Hz), 3.49 [2H, t, J=6HZ), 3.57 (1H, d, J=18Hz) , 4.15 (2H, t, J=9Hz) , 4.31-4.45 (2H, m), 4.94 (1H, d, J-l5Hz), 5.02 (1H, d, J=l5Hz), 5.27 (1H, d, J=5Hz), 5.85 (1H, d, J=5Hz) , 7.95 (1H, s) Preparation38 35 To a suspension of 1,1'-carbonyldiimidazole (2.0 g) in dehydrated chloroform (30 ml) was added a solution of tert-butyl N-(2-hydroxyethyl)carbamate (1.92 g) in dehydrated chloroform (10 ml) under ice-cooling, and the 62 WO 2004/039814 PCT/JP2003/013684 mixture was stirred at room temperature for 1 hour. To the reaction mixture were added N-ethyldiisopropylamine ( 2.2 ml) and 4.5-diamino- 1-methyl -pyrazole sulfuric acid salt (2.58 g) , and the mixture was stirred at room 5 temperature for 17.5 hours. To the reaction mixture were added trityl chloride (3.42 g) and triethylamine (1.25 g) . The mixture was stirred at roara temperature for 2 hours. The reaction mixture was washed with 10% aqueous citric acid solution., brine and saturated 10 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 5% methanol/chloroform to give 4-([2- (tert- 15 butoxycarbonylamino) ethoxycarbonyl ]amino)-5- tritylamino)-l-methylpyrazole (1.91 g) as a solid, 1H-NMR(CDCl3) b 1.46 (9H, s), 2.89 (3H, s), 3.30-3.36 (2H, m) , 4.03-4.07 (2H, m) , 4.37 (1H, brs), 4.75 £1H, br) , 5.42 (1H, br), 7.17-7,30 (l6H, m) 20 Example 21 7 b -[ (Z)-2- (5-Amino-l,2,4-thiadiazol-3-yl)-2- (1- carboxy-l-methylethoxyimin) acetamido] -3- (3-amino-4- [ (2- aminoethoxycarbonyl)amino]-2-methyl-l-pyrazolio)methyl- 3-cephem-4-carboxylate 25 The title compound was obtained from benzhydryl 7 b -((Z)-2-(5-amino-l,2,4—thiadiazol-3-yl)-2-(l-tert- butoxycarbonyl-1-methylethoxyimino)acetamido]-3- iodomethy 1-3-cephem-4-carboxylate and 4-{[2-(tert- butoxycarbony1amino) ethoxycarbonyl] amino)-5- 30 (tritylamino)l-methylpyrazole in the same manner as in Example 20 as an amorphous solid. 1H-NMR(D2O) b 1.53 (3H, s), 1.54 (3H, s), 3.18 (1H, d, J=l8Hz), 3.30-3.38 (2H, m), 3.43 (lH, d, J=15Hz), 3.71 (3H, s), 4.37-4.40 (2S, m), 4.97 (1H, d, J=15Hz), 5.18 35 (1H, d, J=15HZ) , 5.24 (1H, d, J=5Hz), 5.83 (1H, d, J=5Hz), 7.95 (1H, s) Preparation 39 To a solution of 7-amino-2 , 3-dihydro—1H- 63 WO 2004/039814 PCT/JP2003/013684 imidazo[l,2-blpyrazole sulfuric acid salt (1.42 g) and N-ethyldiisopropylamine (2.73 g) in methylene chloride (50 ml) was added N- [2- (tert-butoxycarbonylamino) - acetoxy]succinimide (1.90 g). The mixture was stirred 5 at room temperature for 22 hours. The reaction mixture was washed with saturated aqueous sodium hydrogen carbonate solution, and the organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo, The oily residue was purified by column 10 chromatography on silica gel eluting with 5% methanol/chloroform to give 7-[2-{tert- butoxycarbonylamino)acetyl]amino-2, 3-dihydro-lH- imidazo[1,2-b]pyrazole (1.07 g) as a solid. 1H-NMR(CDCl3) 1.47 [9H, S), 3.89 (2H, d, J=5.5Hz), 3.97 15 (2H, t, J=2.7, 7.3Hz), 4.18 (2H, t, J=7.3Hz), 4.55 (1H, br) , 5.22 (1H, br), 7.16 (lH, s), 7.95 (1H, br) Example 22 To a solution of benzhydryl 7- [ (Z) -- (5-tert- butoxycarbonylamino-1, 2 , 4-thiadiazol-3-yl} -2- (1-tert- 20 butoxycarbony1-1-methylethoxyimino)acetamido]-3- chlormethyl-3-cephem-4-carboxylate (1.0 g) in N,N- dimethylformamide (2.0 ml) was added sodium iodide (181 mg), and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added 7-[2- 25 (tert-butoxycarbonylamino}acetyl]amimo-2,3-dihydro-lH- imidazo [1,2-b]pyrazole (421 mg) , The whole mixture was stirred at 30°C for 3 Hours. To tixe resulting reaction mixture were added ethyl acetate (100 ml) and water (50 ml) . The aqueous layer was separated, and the organic 30 layer was washed with 10% aqueous sodium trifluoroacetate solution and brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (150 ml), and the 35 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 anlsole (1.0 ml) and trifluoroacetic acid (2.0 ml) . The resulting 64 WO 2004/039814 PCT/JP2003/013684 solution was stirred at room temperature for 4 hours and poured into diisopropyl ether (150 ml) . The resulting precipitate was collected by filtration and dried in vacuo to give a crude product (630 mg) , which was 5 purified by preparative HPLC utilising ODS column. The eluate containing a desired product, was concentrated to about 30 ml in vacuo. The concentrate was adjusted to about pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical 10 Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo and lyophilized to give 7[ (Z) -2-(5-amino-l, 2,4-thiadiazol- 3-yl} -2- (1-carboxy-l-methylethoxyimino) acetamido]-3- [7- (2-aminoacetamido) -2 , 3-dihydro-5- ( 1H-imidazo [1,2- 15 b] pyrazolio) ]methyl-3-cephem-4-carboxylate (20. 3 mg) as an amorphous solid. lH-NMR(D2O) 1.51 (3H, s) , 1.52 (3H, s) , 3.26 (2H, t, J=18Hz), 3.54 (2H, d, J=18Hz), 3.97 (2H, s), 4.16 (2H, t, J=9Hz) , 4.35 (1H, q, J=9Hz) , 4.44 (1H, q, J=9Hz), 4.97 20 (2H, d, J=15HZ), 5.04 (2H, d, J=l5Hz), 5.25 (lH, d, J=4Hz), 5.84 (1H, d, J=4Hz) , 8.10 (lH, s) Preparation 40 To a suspension of 4,5-diamino-l-(2- hydroxyethyl)pyrazole sulfuric acid salt (1.20 g) and N- 25 [2-(tert-butoxycarbonylamino) acetoxy] succinimide [l.35 g) in methylene chloride (20 ml) was added N- ethyldiisopropylamine (2.1 ml) under ice-cooling, and the mixture was stirred at room temperature for 17 hours. The reaction mixture was washed with water (4 0 ml) , 30 saturated aqueous sodium hydrogen carbonate solution (40 ml) and brine (40 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The oily residue was purified by column chromatography on silica gel eluting with 10% 35 methanol/chloroform to give 5-amino-4- [2- (tert- butoxycarbonylamino) acetyl ] amino-1- ( 2- hydroxyethyl)pyrazole (l.20 g) as a solid- 1H-NMR(CDCl3) 1.46 (9H, 5), 3.89-3.90 (4H, m), 4.00- 65 WO 2004/039814 PCT/JP2003/013684 4.04 (2H, m, 4.26 (2R, br) , 5.51 (1H, br) , 7.17 (1H, s) , 8.06 (1H, br) Example 23_ 7 b - [ (Z) -2-(5-Amino-l,2,4-thiadiazol-3-yl) -2- (1- 5 carboxy-l-methylethoxyimino) acetamido]-3- [3-amino-4- (2- aminoacatamido) -2- (2-hydroxyethyl) -l-pyrazolio]methyl-3- cephem-4-carboxylate The title compound was obtained from benzhydryl 7 b -(Z)-2-(5-tert-butoxycarbonylamino-l,2,4-thiadiazol- 10 3-yl)-2-(1-tert-butoxycarbonyl-l-methylethoxyimino)- acetamido]-3—chloromethy1—3-cephem-4-carboxylate and 5— amino—4-[2- (tert-butoxycarbonylamino) acetyl] amino-1- (2- hydroxyethyl) pyrazole in the same manner as in Example 22 as an amorphous solid. 15 1H-HMR(D2O) 1.52 (6H, s), 3.15 (2H, d, J=l8Hz), 3.48 (2H, d, J=18Hz) , 3,88 (1H, dt, J=16Hz) 4.02 (2H, s) , 4.42 (1H, dt, J=16.5Hz), 5.07 (2H, d, J=15HZ), 5.13 (2H, d, J=15Hz), 5.27 (lH, d, J=5Hz), 5.84 {1H, d, J=5Hz), 8.09 {1H, s) 20 Preparation 41 To a solution of 3-amino-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrimidine sulfuric acid salt (2.96 g) and N-ethyldiisopropylamine (2.59 g) in methylene chloride (70 ml) was added N-[2- (tert— 25 butoxycarbonylamino)acetoxy]succinimide (2.72 g). The mixture was stirred at room temperature for 14 hours. The reaction mixture was washed with saturated aqueous sodium hydrogen, carbonate solution. The organic layer was dried over anhydrous magnesium sulfate, filtered and 30 concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with 6% methanol/chloroform to give 3-[2-(tert- butoxycarbonylamino) acetyl] amino-4 ,5,6,7- tetrahydropyrazolo[1,5—alpyrimidine (2.4 g) as a solid. 35 1H-NMR(CDCl3) 1.46 (9H, s) , 2.08-2.12 (2H, m) , 3.29- 3.32 (2H, m), 3.90 (2H, br), 4,07 (2H, t, J=6.0Hz), 5.00 (1H, br), 5,38 (1H, br), 7.12 (lH, s), 8.11 (1H, br) Example 24 66 WO 2004/039814 PCT/JP2003/013684 7 b -[(Z) -2-(5-Amino-l,2,4-thiadiazol-3-yl)-2- (1- carboxy-l-methylethoxyimino) acetamido ] -3- [3- (2- aminoacetamido) -4,5,6, 7-tetrahydro-l-pyrazolo [1,5- alpyrimidinio] methy1-3-cephem-4-carboxylate 5 The title compound was obtained from benzhydryl 7 b - [ (Z) -2- (5-tert-butoxycarbonylamino-l, 2 ,4-thiadiazol- 3-yl) -2- (1-tert.-butoxycarbonyl-l-methylethoxyimino)- acetamido]-3-chloromethyl-3-cephem-4-carboxylate and 3- [2- (tert-butoxycartibonylamino) acetyl) amino-4 ,5, 6,7- 10 tetrahydropyrazolo [1, 5-a ] pyrimidine in the same manner as in Example 22 as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s) , 1.53 (3H, s), 2.05-2.25 (2H, m) , 3.21 (2H, d, J=18Hz) , 3.45 (2H, d, J=18Hz) , 3.30- 3.45 (2H, m) , 4.00 (2H, s), 4.10-4.25 (2H, m) , 4.92 (2H, 15 d, J=15Hz), 5.17 (2H, d, J=15Hz) , 5.24 (1H, d, J=5Hz) , 5.84 (1H, d, J=5HZ), 7.97 (1H, s) Preparation 42 To a solution of 3-amino-4 , 5 , 6 ,7- "tetrahydropyrazolo [1 , 5-a] pyrimidine sulfuric acid salt 30 (4.44 g) and N-ethyldiisopropylamine (3.88 g) in methylene chloride (100 ml) was added N-[3-(tert- butoxycarbonylamino)propionyloxy) succinimide (4.29 g) . The mixture was stirred at room temperature for 6 hours. The reaction mixture was washed with saturated aqueous 25 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 5% methanol/chloroform to give 3-[3-(tert- 30 butoxycarbonylamino)propiony1]amino-4,5,6,7- tetrahydropyrazolo(1,5-alpyrimidine (3.67 g) as an oil. 1H-NMR(CDCl3) 1.43 (9H, s) , 2.08-2.13 (2H, m) , 2.52 (2H, t, J=6.0Hz), 3.32 (2H, t, J=5.0Hz), 3.43-3.46 (2H, m) , 4.07 (2H, t, J=6.0Hz, 5.12 (1H, br), 5.23 (1H, br), 35 7.13 (1H, s) , 7.97 (1H, br) Example 25 7 b - [ (Z) -2- (5-Amino-l,2 , 4-thiadiazol-3-yl) -2- (1- carboxy-l-methylethoxyimino) acetamido] -3- [3- (3- 67 WO 2004/039814 PCT/JP2003/013684 aminopropionamido) -4,5,6, 7-tetrahydro-l-pyrazalo [1,5- a] pyrimidinio] methyl-3-cephem-4-carboxyylate The title compound was obtained from benzhydryl 7 b -[ (Z) -2- (5-tert-butoxycarbonylamino-l ,2,4-thiadiazol- 5 3-yl) -2- (1-tert-butoxycarbonyl-1-methylethoxyimino)- acetamido]-3-chloromethyl-3-cephem-4-carboxylate and 3- [3- (tert-butoxycarbonylamino) propionyl]amino-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrimidine in the same manner as in Example 22 as an amorphous solid. 10 1H-NMR(D2O) 1.51(3H,s), 1.52 (3H, s),2.05-2.25 (2H, m) ,2.85 (2H,t, J=7Hz),3.20 (2H, d,J=l8Hz),3.44 (2H, d, J=18Hz), 3.30-3.45 (2H,m),3.31 (2H, t, J=7Hz), 4.05-4.20 (2H, m), 4.91 (2H,d,J=16Hz), 5.16(2H,d, J=16HZ) ,5.23 (1H,d, J=5Hz), 5.84 (1H, d, J=5Hz) , 7.92 15 (1H, s) Preparaton 43 To a solution of 5-amino-methylpyrazole (100 g) in water (700 ml) were added concentrated hydrochloric acid (86 ml) and sodium nitrite (63.9 g) in water (200 20 ml) at a temperature below 10°C The reaction mixture was stirred at 5°C for 30 minutes. The precipitated solid was collected by filtration and dried to give 5- amino-l-methyl-4-nitrosopyrazole (117 g). 1H-NMR(DMSO-d6) 3.52 and 3.59 (3H, s), 7.22 and.8.51 25 (1H, s) , 8.17 and 8.51 (1H, brs) Preparation 44 To a suspension of 5-amino—1-methyl—4- nitrosopyrazole.(117 g) were added sulfuric acid (91 g) and 10% palladium on carbon (58 g). The mixture was 30 hydrogenatad under balloon pressure for 10 hours. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. To the concentrate was added isopropyl alcohol (2.3 L), and the mixture was stirred for 1 hour. The precipitated solid, was collected by 35 filtration and dried to give 4,5-diamino—1- methylpyrazole sulfuric acid salt (158 g). 1H-NMR(D2O) 3.74 (3H, s), 7.80 (1H,s) Preparation 45 68 WO 2004/039814 PCT/JP2003/013684 A solution of 4, 5-diamino-1-methylpyrazole sulfuric acid salt (158 g) in water (l.l L) was neutralized to pH 6.9 with 4N aqueous sodium hydroxide solution, and dioxane (474 ml) was added to this 5 solution. To the resulting mixture was added dropwise phenyl chloroformate (124 g) maintaining pH of the mixture at 6.9 with 4N aqueous sodium hydroxide solution at a temperature below 10°C. The reaction mixture was stirred for 1 hour . The precipitated solid was 10 collected by filtration and dried to give 5-amino-l- methyl-4-phenoxycarbonylaminopyrazole (155 g) . 1H-NMR(DMSO-d6) 3.52 (3H ,s) , 5.00 (2H, brs) , 7.10-7.50 (6H, m) , 8.93 (1H, brs) Preparation 45 15 To a suspension of 5-amino-l-methyl-4- phenoxycarbonylaminopyrazole (153.8 g) in tetrahydrofuran (1 L) were added triethylamine (67 g) and triphenylmethyl chloride (185 g) at room temperature. The mixture was stirred for 6.5 hours. To the reaction 20 mixture was added heptane (2.6 L) , and the mixture was stirred for 1 hour. The precipitated solid was collected by filtration and washed with heptane- diisopropyl ether (1:1) . The crude solid was suspended in water (3 L), and the suspension was stirred for 1 25 hour. The solid was collected by filtration and dried to give l-methyl-4-phenoxycarbonyl-5- triphenylmethylaminopyrazole (253.6 g) . 1H-NMR(DMSO-d6) 2.74 (3H, s) , 5.57 (1H, brs) , 7.00-7.50 (21H, m), 8.12 (1H, brs) 30 Preparation 47 To a suspension of l-methyl-4- phenoxycarbonylamino-5-triphenymethylaminopyrazole (253.6 g) in N,N-dimethylformamide (l.5 L) were added triethylamine (59.5 g) and tert-butyl N-(2- 35 amino thyl) carbamate (94.2 g) in N,N -dimethylformamide (254 ml) . The mixture was stirred for 5 hours and poured into water (10.6 L). The slurry was stirred for 1 hour. The precipitated solid was collected by 69 WO 2004/039814 PCT/JP2003/013684 filtration and dried to give a crude product. The crude product was suspended in N,N-dimethylformamide, and the suspension was heated under reflux for 20 minutes. The suspension was cooled to ambient temperature over 4 5 hours. The solid was collected by filtration, washed with acetonitrile and dried to give 4-[N-(2-tert- butoxycarbonylaminoethyl) carbaramoylamino] -l-methyl-5- triphenylmethylaminopyrazole (261.2 g) . 1H-NMR(DMSO-d6) 2.69 (3H, s), 2.30-3.05 (4H, m) , 5.69 10 (1H, brs), 5.51-6.01 (1H, m), 6.74-6.81 (1H, m) , 6.87 (1H, brs) , 7.00 (1H, s), 7.10-7.30 (15H, m) Preparation 4 8 To a solution of (Z)-2-[5 -amino-1,2,4-thiadiazol- 3-yl) -2- (l--tert-butoxycarbonyl-l- 15 methylethoxyimino) acetic acid (319 g) in N,N- dimethylacetamide (1.5 L) were added potassium carbonate (113 g) and methanesulfonyl chloride (126 ml) under ice- cooling. The mixture was stirred at 10°C for 2 hours. The reaction mixture was added to a mixture of ethyl 20 acetate and water. The organic layer was washed with water and brine to give an activated acid solutiont On the other hand, a suspension of 4-methoxybenzyl 7- amino-3-chloromethyl-3-cephem- 4-carboxy late hydrochloride (3 0 0 g) in a mixture of water (1 L) and 25 ethyl acetate (1 L) was adjusted to pH 6 with triethylamine under ice-cooling. To the resulting mixture was dropwise added the above obtained activated acid solution, at 10oC under stirring. Stirring was continued at 5-10°C for 1.5 hours keeping pH of the 30 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 L), and the resulting precipitate was collected by filtration and dried to give 4- 35 methoxybenzyl 7-[(Z)-2-[5-amino-l,1,4-thiadiazol-3-yl)- 2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamidol- 3-chloromethyl-3-cephem-4-carboxylate (495.7 g) . 1H-NMR(DMSO-d6) 1.39 (9H, s), 1.44 (6H, s), 3.45-3.70 70 WO 2004/039814 PCT/JP2003/013684 (2H, m) , 3.76 (3H, s) , 4.46 and 4.54 (1H, ABq, J=16HZ), 5.10-5.28 (2H+lH, m) , 5.90 (1H, dd, J=4 .9, 8.5Hz), 6.94 (2H, d, J=8.7Hz), 7.36 (2H, d, J=8.7Hz), 8.18 (2H, brs) , 9.52 (1H, d, J=5.5Hz) 5 Example 26 To a solution of 4-methoxybenzyl 7-[(Z)-2- (5- amino-1, 2 ,4-thiadiazol-3 -yl) -2 - (l-tert-butoxycarbonyl-l- methylethoxyimino) acetamidol -3-chloromethyl-3-cephem-4- carboxylate (150 g) in N,N-dimethylformamide (4 00 ml) 10 was added 1 ,3-bis (trimethylsilyl) urea (225 g) and the mixture was stirred for 30 minutes. Potassium iodide (51.2 g) was added to this solution, and the mixture was stirred for 30 minutes, 4-[N- (2-tert-Butoxycarbonylaminoethyl) - 15 carbamoylamino] -1 -methyl-5-triphenylmethylaminopyrazole (147 g) was dissolved in N,N-dimethylformamide (650 ml) at 78oC and the solution was cooled to 45°C, The solution was added to the solutian of 7-[ (Z)-2-(5- amino-1, 2 ,4-thiadiazol-3-yl) -2- (l-tert-butoxycarbonyl-l- 20 methylethoxyimino) acetamido] -3-chloromethy 1-3-cephem-4- carboxylate obtained above. The reaction mixture was stirred at 35°C for 18. 5 hours and poured into a mixture of ethyl acetate (21), water (1.8 L) and 20% aqueous sodium chloride solution (150 ml) , The organic layer 25 was washed with a mixture of 10% aqueous sodium thiosulfate solution (375 ml) and 20% aqueous sodium chloride solution (375 ml). The organic layer was washed successively with 10% aqueous sodium trifluoroacetate solution three times (750 ml x 3) and 30 20% aqueous sodium chloride solution (750 ml) . The organic layer was concentrated in vacuo , and the precipitated 4- [N- (2-tert-butoxycartbonylaminoethyl) - carbamoylamino]-l-methyl-5-triphenylmethylaminopyrazole was filtered off. The filtrate was further concentrated 35 in vacuo to a volume of approximately 6 00 ml. This solution was added to diisopropyl ether and the suspension was stirred for 1 hour. The resulting solid was collected by filtration and dried. The solid was 71 WO 2004/039814 PCT/JP2003/013684 dissolved in dichloromethane (669 ml) . To the solution were added anisole (223 ml) and trifluoroacetic acid (669 ml) . The reaction mixture was stirred for 4 hours and poured into diisopropyl ether. Tne resulting solid 5 was collected by filtration and dried. This solid was suspended in water, and pH of the suspension was adjusted to 1 with aqueous ammonia solution at a temperature below 10°C. The resulting precipitate was filtered off. The filtrate was acidified to pH 1 with 10 concentrated hydrochloric acid at a temperature below 10°C, and the resulting precipitate was filtered off. The filtrate was chromatographed on Diaion® HP-20 (11 L) eluting with 20% aqueous Z-propanol. The eluate was concentrated to about 3.5 L in vacuo, and 2M sulfuric 15 acid (51 ml) was added. The mixture was lyophilized to give a crude product (72.2 g) . The crude product (3 g) was purified by preparative HPLC utilizing ODS column. The eluate containing a desired product wag concentrated in vacuo. 20 The concentrate was adjusted to about pH 1 with concentrated hydrochloric acid and chromatographed on. Diaion® HP-20 (400 ml) eluting with 20% aqueous 2- propanol. The eluate was concentrated to about 73 ml in vacuo, and 2M sulfuric acid (1.5 ml) was added. The 25 mixture was further evaporated to a volume of approximately 12.5 ml, and water (6 ml) was added. After addition of seed crystals (10 mg), which resulted in the precipitation of a white solid, the mixture was stirred at room temperature for 1 hour. The mixture was 30 further stirred at 5°C for 13 hours. 2-Propanol £20 ml) as added at 5°C over 20 minutes , and the slurry was tirred at room temperature for 4 hours. 2-Propanol (20 l) was added over 30 minutes, and the slurry was tirred at room temperature for 3 hours. The 35 recipitated crystals were collected by filtration and ried to give 70-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)- 2—(l-carboxy-l-methylethoxyimiiio)acetamido]—3-(3-amino- 4- [H- (2-aminoethyl) cacbamoylamino] -2-methyl-l- 72 WO 2004/039814 PCT/JP2003/013684 pyrazolio)methyl-3-cephem-4-carboxylic acid hydrogen sulfate (1.51 g) as crystals. 1H-NMR(D2O) 1.61 (6H, s), 3.10-3.55 (6H, m), 3.71 (3H, 3), 5.02 and 5.23 (2H, ABq, J=16.7Hz), 5.25 (1H, d, 5 J=4.9Kz), 5.87 (1H, d, J=4.9Hz), 7.91 (lH, s) Preparation 49 A suspension of 4 , 5-diamino-l-methylpyrazole sulfuric acid salt (20 g) in triethylamine (29.2 ml) was stirred at 0°C for 10 minutes. A mixture of acetic 10 anhydride (9.87 ml) and formic acid (7.96 ml) was stirred at 4O°C for 30 minutes, cooled, to 0°C, and added dropwise to the above solution at 0°C. The whole mixture was stirred at 0°C for 2 hours. To the mixture was added brine, and the whole mixture was extracted 15 with tetrahydrofuran. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure to give crude N—(5-amino-l-methyl-lH—pyrazol-4- yl) formamide, whicii was used in the next step without further purification. 20 Preparation 50 The crude product of N-(5-amino-l-methyl-lH- pyrazol—4-yl) formamide (13.33 g) was dissolved iri N,N- dimethylformamide (130 ml) . To the solution were added. trityl chloride (29.2 g) , triethylamine (66.3 ml) and 4- 25 dimethylaminopyridine (465 mg) , and the mixture was stirred at 60oC for 5 hours. To the reaction mixture was added water, and the whole mixture was extracted with ethyl acetate. The organic layer was washed with water and brine, and dried over magnesium sulfate. The 30 solvent was evaporated under reduced pressure to give a white solid. The solid was triturated with ethyl acetate/diisopropyl ether (1:1) to give N— [1-methyl-5— (tritylamino)-lH-pyrazol-4-yl] formamide (19.18 g) . The NMR spectrum of this compound indicates the existence of 35 its rotamer. lH-NMR(DMSO-d6) 2.76 and 2.92 (3H, s), 5.56 and 5.84 (1H, s), 7.0-7-4 (16H, m), 7.66 [1H, d, J=1.7Hz), 8.3- 8.4 (1H, m) 73 WO 2004/039814 PCT/JP2003/01368 ESI-MS: m/z =405.2(M+Na) + Preparation 51 To a solution of N- [l-methyl-5- (tritylamino) -1H- pyrazol-4-yl] formamide (3.825 g) in N,N- 5 dimethyl formamide (66 ml) was added sodium hydride (264 mg, 60% oil suspension) under a nitrogen atmosphere at 0oC under stirring. The mixture was stirred at 0oC for 15 minutes. To the mixture were added tert-butyl N-(3- bromopropyl) carbamate (2.62 g) in N,N-dimethylformamide 10 (10 ml) and sodium iodide (1.65 g) , The mixture was warmed to room temperature and stirred for 2 hours. 10% Aqueous potassium hydrogen sulfate solution (5 ml) was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with water and brine, and 15 dried over magnesium sulfate. The solvent was evaporated under reduced pressure, and the residue was chromatographed on silica gel eluting with methylene chloride/ethyl acetate (4:1) to give tert-butyl 3-(N- formyl-N-[l-methyl-5-(tritylamino)-lH-pyrazol-4- 20 yl]amino)propylcarbamate (2.714 g) . The NMR spectrum of this compound indicates the existence of its rotamer. 1H-NMR(DMSO-d6) 1.37 and 1-39 (9H, s) , 2.6-2.9 (6H, m) , 2.89 (3H, s), 5.34 and 6.01 (1H, s), 6.6-6-3 (lH, m) , 7.0-7.4 (l5H, m), 7.5-7.6 (1H, m), 7.95 (1H, s) 25 ESI-MS: m/z-5 62.3(M+Na)+ Example 27 7 b -[(Z)-2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-(1- carboxy-1 -methylethoxyimino) acetamido] -3- (3 -amino-4- [N- (3-aminopropyl) -N-formylamino] -2-methyl-1- 30 pyrazolio)methyl-3-cephem-4-carboxylate The title compound was obtained from benzhydryl 7 b -[ (Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(l-tert- butoxycarbonyl-l-methylethoxyimino) acetamido] -3- iodomethyl-3-cephem-4-carboxylate and tert-butyl 3-(N- 35 formyl-N-[l-methyl-5-(tritylamino)-lH-pyrazol-4- yl] amino)propylcarbamate in the same manner as in Example 1. The NMR spectrum of this compound indicates the existence of its rotamer. 74 WO 2004/039814 PCT/JP2003/013684 lH-NMR(D2O) 1.53 (6H, s), 1.7-2.1 (2H, m) , 2.9-3.9 (9H, m) ,4.97 and 5.20 (2H, ABq, J=15.2Hz), 5.26 (1H, d, J=4.8HZ), 5.64 (1H, d, J=4.8Hz), 8.0-3.3 (2H, m) Example 28 5 To a suspension of 7-[(Z)-2-(5-amino-l,2,4- thiadiazol-3-yl) -2- (l-carboxy-1-methylethoxyimino)- acetamido]-3—(3—amino—4- [N- (3-aminopropyl)-N- formylamino]—2-methyl—1-pyrazolio]methyl-3-cephem—4- carboxylate (140 mg) in methanol (2.6 ml) was added 10 concentrated hydrocloric acid (0.176 ml) at room temperature, and the mixture was stirred for 6.5 hours. To the reaction mixture was added sodium hydrogen carbonate (177 mg), and the mixture was purified by preparative HPLC (ODS column, acetonitrile/phosphate 15 buffer (pH 7)=5:95). The eluate containing a desired product was evaporated to remove acetonitrile, acidified with diluted hydrochloric acid and chromatographed on Diaion® Hp-20 eluting with 20% aqueous 2-propanol. The eluate was concentrated under reduced pressure and 20 lyophilized to give 7-[(Z)-2-[5-amino-l,2,4-thiadiazol- 3-y1)-2-(1-carboxy-l-methylethoxyimino) acetamido] -3-(3- amino-4—[ (3-aminopropyl) amino] -2- methyl—1— pyrazolio}methyl-3-cephem-4-carboxylate (39 mg), 1H-NMR(D2O) 1.52-1.54 (6H, m), l.95 (2H, tt, J=7.3Hz, 25 7.3Hz), 3.0-3.2 (4H, m) , 3.16 and 3.38 (2H, ABq, J=17,7Hz), 3.68 (3H, s) , 4.89 and 5.11 (2H, ABq, J=15.6Hz), 5.22 (1H, d, J=4.3Hz), 5.83 (1H, d, J=4.8Hz), 7.59 (1H, s) ESI-MS: m/z=636.3(M-H)- 30 Preparation 52 tert-Butyl 2-{N-formyl-N-[1-methyl-5- (tritylamino) -lH-pyrazol-4-yl] amino] ethylcarbamate The title compound was obtained from N-[1-methyl- 5-tritylamino)-lH-pyrazol-4-yl]formamide and tert-butyl 35 N- (2-bromoethyl) carbamate in the same manner as in Preparation 51. IR(KBr) 1709, 1670, 1170, 704 cm-1 1H-NMR(DMSO-d6) 1.35 and 1.3 6 (9H, s), 2.65 and 2.7 5 75 WO 2004/039814 PCT/JP2003/013684 (3H, s), 2.73-2.90 (4H, m), 5.45 and 6.02 (1H, s), 6.78 and 6.33 (1H, t-like), 7.05-7.30 (15H, m), 7.31 and 7.57 (1H, s) ESI-MS: m/z=426.3 (M+H+) , 548.3(M+Na+) 5 Example 29 7 b -[ (Z)-2-(5-Amino-l,2, 4-thiadiazol-3-yl) -2-(1- carboxy-l-methylethoxyimino) acetamido] - 3-{3-amino-4- [N- (2-aminoethyl) - N-forylamino] -2-methyl-1- pyrazolio} methy1-3 -cephem-4-carboxylate 10 The title compound was obtained from benzhydryl 7 b [ (Z)-1-(5- amino-l,2,4-thiadiazol-3-yl)-2- (l-tert- butoxycarbonyl-l-methylethoxyimino)acetamido]-3- chloromethyl -3-cephem-4-carboxylate and tert-butyl 2-{N- formyl-N-[l-methyl-5-(tritylamino)-lH-pyrazol-4- 15 yl]amino}ethylcarbamate in the same manner as in Example 1. IR(KBr) 1770, 1675, 1653, 1597 cm-1 1H-NMR(DMSO-d6) 1.53 (6H, s), 3.12-3.78 (4H, m) , 3.77 and 3,78 (3H, s) , 3.86-3.96 (2H, m) , 5.00 and 5.19 (2H, 20 ABq, J= 15.2Hz), 5.28 (1H, d, J=4.8Hz), 5.86 (1H, d, J=4.8Hz), 8.15 and 8.18 (1H, s), 8.19 and 8.33 (lH, s) ESI-MS: m/z=652.2(M+H+) Example 30 7 b -[ (Z) -2-(5-Amino-l, 2 , 4-thiadiazol-3-yl)-2- (1- 25 carboxy-l-methylethoxyimino ) acetamido] -3 - [3-amino-4- [ (2- aminoethyl ) amino ] -2-methyl -1- pyrazolio] methyl -3-cephem- 4-carboxy1ate The title compound was obtained from 7-[ (Z)-2-(5- amino-1,2 , 4-thiadiazol-3-yl) -2- (1-carbozy-l- 30 methylethoxyimino)acetamido]-3-(3-amino-4-[N-(2- aminoethyl) -N-formylamino)-2-methyl-l-pyrazolio)metliyl- 3-cephem-4-carboxylate in the same manner as in Example 28 . IR(KBr) 1770, 1651, 1593 cm-1 35 1H-NMR(DMSO-d6) 1.53 (3H, s) , 1.59 (3H, s), 3.13-3.26 (4H, m), 3.26 and 3.39 (2H, ABq, J=17.8Hz), 3.68 (3H, s), 4.87 and 5.11 (2H, ABq, J=15.7Hz), 5.25 (1H, d, J=4.8Hz), 5.84 (1H, d, J=4.8Hz), 7.63 (1H, s) 76 WO 2004/039814 PCT/JP2003/013684 ESI-MS: m/z=622,2(M-H-) Preparation 53 To a suspension of l-methyl-lH-pyrazole-4, 5- diamine sulfate (86 g) in tetrahydrofuran (1.3 L) was 5 added triethylamine (117 ml), and then (2S)-4-[ (tert- butoxycarbonyl) amino]-2-hydroxybutanoic acid (82.5 g) was added to the mixture. To the mixture were added 1- hydroxybenzotriazole (58.3 g) and N-(3- dimethylaminopropyl) -N' -e.thylcarbodiimide hydrochloride 10 (82.7 g) under ice-coaling, The reaction mixture was stirred at room temperature for 8 hours. To the reaction mixture were added ethyl acetate (l.3 L) , saturated aqueous sodium hydrogen carbonate solution and sodium chloride, and the mixture was stirred for 30 15 minutes. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (1.0 L) six times. The extra was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel column 20 chromatography eluting with ethyl acetate/tetrahydrofuran (l/l) to give tert-butyl [(3s)- 4- [ (5-amino-l-methyl-lH-pyrazol-4-yl) amino] -3-hydroxy-4- oxobutyl} carbamate (69. 5g). 1H-NMR(CDC13) 1.43 (9H, s), 1.6-1.9 (1H, m) , 1.9-2.2 25 (1H, m), 3.1-3.3 (1H, m) , 3.3-3.5 (lH, m) , 3.65 (3H, s) , 4.20 (1H, dd, J=3.6, 6.6Hz), 4.7-5.3 (4H, m), 7.24 (1H, s) , 8.58 (1H, s) [b D(c=l.05, CHC13]-27.06° Preparation 54 30 To a solution of tert-butyl [(3S)-4-[{5-amino-l- methyl-lH-pyrazol-4-yl)amino]-3-hydroxy-4- oxobutyl}carbamate (68.-51 g) in N,N-dimethylformamide (350 ml) was added chlorotriphenylmethane (67 g). To the mixture was dropwise added triethylamine (6 7 ml) . 35 The mixture vas stirred at room temperature for 12 hours. The reaction mixture was dissolved in dichloromethane (2 L) . The solution was washed successively with water and brine. The extract was dried over anhydrous magnesium 77 WO 2004/039814 PCT/JP2003/013684 sulfate, filtered and concentrated in vacuo. The residue was triturated with acetonitrile and dried in vacuo to give tert-butyl [(3S)-3-hydroxy-4-{[l-methyl-5- (tritylamino)-lH-pyrazol-4-yl]amino]-4- 5 oxobutyl)carbamate (64 g). 1H-NMR(CDCl3) 1.46 (9H, s), 1.3-1.6 (1H, m) , 1.8-2.1 (1H, m), 2.95 (3H, s), 2.9-3.2 (1H, m) , 3.3-3.6 (1H, m), 3.95 (1H, m), 4.53 (1H, d, J=4.5Hz), 4.74 (1H, s), 4.92 (1H, brs), 7.1-7.3 (15H, m), 7.39 (1H, s), 7.73 (1H, s) 10 ESI-MS: m/z=6 38 .2(M+H+Na+) [b]20D{c=l.025, CHC13)=-36.5° Example 31 To a solution of 4-methoxybenzyl 7-[ (Z)-2—(5- amino-1,2,4-thiadiazol-3-yl) -2- (l-tert-butoxycarbonyl-l- 15 methylethoxyimino) acetamido] -3-chloromethyl-3-cephem-4— carboxylate (130 g) in N ,N- dimethyl formamide (400 ml) was added 1,3-bis(trimethylsilyl)urea (195 g) , and the mixture was stirred at room temperature for 30 minutes. To the solution was added potassium iodide (44.4 g), and 20 the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added tert-butyl ((3S) -3-hydroxy-4-{[l-methyl-5-(tritylamino)-lH-pyrazol- 4-yl]amino)-4-oxobutyl}carbamate (106 g), and the whole mixture was stirred at 35°C for 22 hours. To the 25 reaction mixture was added ethyl acetate (1.7 L), and the mixture was washed successively with water (1.6 L), 10% aqueous sodium trifluoroacetate solution (650 ml x 3) and brine (650 ml), dried over magnesium sulfate and filtered. The filtrate was concentrated to about 1 L in 30 vacuo. The concentrate was poured into diisopropyl ether (3 L), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the solid in methylene chloride (660 ml) were added anisole (220 ml) and trifluoroacetic acid (660 ml) . 35 The resulting solution was stirred at room temperature for 4 hours and poured into diiaopropyl ether (7 L) . The resulting precipitate was collected by filtration and dried in vacuo to give a cruds product 78 WO 2004/039814 PCT/JP2003/013684 (156.2 g). The crude product was dissolved in water (3.5 L). The solution was adjusted to about pH 3 with concentrated hydrochloric acid and chromatographed cm Diaion® HP-20 (Mitsubishi Chemical Corporation) eluting 5 with 20% aqueous 2-propanol. The eluate was concentrated to about 1.5 L in vacuo, and 2M aqueous sulfuric acid solution (33.18 ml) was added. The mixture was lyophilized. The lyophilised product (40 g) was dissolved in phosphate buffer (pH 7} and purified by 10 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 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical 15 Corporation) eluting with 10% aqueous 2-propanol. The eluate was concentrated to about 1 L in vacuo , and 2M aqueous sulfuric acid solution was added (13.59 ml). The resulting solution was lyophilized to give 7[(Z)- 2- (5-amino-l, 2 ,4-thiadiazol-3-yl) -2- (1-carboxy-l- 20 methylethoxyimino) acetamido] -3- {3-amino-4- [ [(2S)-4- amino-2-hydroxybutanoyl) amino] -2-methyl-l- pyrazolio)methyl-3-cephem-4-carboxylic acid hydrogen sulfate (20.32 g) as an amorphous solid, 1H-NMR(D2O) 1.61 (6H, s), 1.9-2.4 (2H, m), 3.20 (1H, d, 25 J=17.6Hz), 3.0-3.3 (2H, m), 3.45 (1H, d, J=l7.6Hz, 3.74 (3H, s), 4.47 (1H, dd, J=4, 6.3Hz), 5.06 (1H, d, J=15.7Hz), 5.25 (1H, d, J=4.8Hz), 5.28 (1H, d, J=15.7Hz), 5.87 (1H, d, J=4.8Hz), 8.07 (1H, s) Preparation 55 30 To a suspension of l-methyl-N5-trityl-lH-pyrazole- 4,5-diamine (1.60 g) in ethanol (50 ml) were added triethylamime (0.627 ml) and diethyl squarate (0.858 ml), and the mixture was stirred at room temperature for 22 hours. To the reaction mixture were added ethyl acetate 35 (200 ml) and hexane (100 ml), and the solution 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 79 WO 2004/039814 PCT/JP2003/013684 vacuo. The crystalline residue was washed with diethyl ether and dried in vacuo to give 3—ethoxy-4-{[1-methyl- 5-(tritylamino)-lH-pyrazol-4-yl]amino}-3-cyclobutene- 1,2-dione (1.45 g) as a solid. 5 1H-NMR(CDCl3) 1.42 (3H, br) , 2.99 (3H,s) , 4.41 (1H, brs), 4.69 (2H, q, J=7.2Hz), 6.40 (1H, br), 7.13-7.35 (16H,m) Preparation 56 To a suspension of tert—butyl 2- 10 aminoethylcarbamate (288 mg) and 3-ethoxy-4-{[l-methyl- 5-{tritylamino)-1H-pyrazol—4-yl]amino)-3-cyclobutene- 1,2-dione (718 mg) in ethanol (20 ml) was added triethylamine (0.209 ml), and the mixture was stirred under reflux for 4 hours. To the reaction mixture were 15 added diethyl ether and hexane. The crystalline precipitate was collected by filtration and dried in vacuo to give tert-butyl 2-[{2-([l-methyl-5- tritylamino)-lH-pyrazol-4-yl]amino)-3,4-dioxocyclobut- 1-en-l-yl)amino]ethylcarbamate (830 mg) as a solid. 20 1H-NMR(CDCl3) 1.40 (9H,s) 3.07-3-28 (5H, m) , 3.38- 3.67 (2H, m), 4.53-4.84 (lH, br), 4.84 (1H, br), 7.15- -7.22 (6H, m) , 7.23 (lH, s) , 7.22-7.34 (9H, m) Example 32 To a solution of benzhydryl 7-[(Z) -2- (5-amino- 25 1,2,4—thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-l- methylethoxyimino) acetamido] -3-iodomethyl-3-cephem-4- carboxylate (9 01 mg) in N,N-dimethylformamide (l.8 ml) was added N— (trimethylsilyl) acetamide (720 mg) , and the mixture was stirred at room temperature for 1 hour. To 30 the reaction mixture was added a solution of tert—butyl 2-[(2-{(l-methyl-5-(tritylamino)-lH-pyrazol-4-yl]amino}- 3 , 4-dioxocyclobut-l-en-l—yl) amino] ethylcarbamate (682 mg) in N,N-dimethylformamide (6.3 ml) , and the whole mixture was stirred at 35-40°C for 7 hours. To the 35 resulting reaction mixture was added ethyl acetate, and the precipitate was filtered off. The filtrate was washed successively with water and brine, dried over anhydrous sodium sulfate and filtered. The filtrate was 80 WO 2004/039814 PCT/JP2003/013684 concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (80 ml) , and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in 5 methylene chloride (2.6 ml) were added anisole (0.88 ml) and trifluoroacetic acid (2.6 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether (80 ml) . The resulting precipitate was collected by filtration and dried in 10 vacuo to give a crude product (580 mg) , 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 with concentrated hydrochloric acid and 15 chromatographed on Diaion 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 3-{[3-amino-4-({2-[(2- aminoethyl) amino]-3,4-dioxo-l-cyclobuten-l-yl)amino)-2- 20 methyl-1-pyrazoliolmethyl}-7-[(Z) -2- (5-amino-l ,2 , 4- thiadiazol-3-yl) -2- (l-carboxy-l- methylethoxyimino) acetatamido] -3-cephem-4-carboxylate (22 mg) as an amorphous solid. 1H-NMR(D2O) 1.53 (3H,s) 1.54 (3H, s), 3.26-3.36 (1H, 25 ml, 3.27 (2H, t, J=5.7Hz) , 3.58-3.69 (1H,m), 3.74 (3H, s), 3.86-4.03 (2H, m), 4.93 (lH, d, J=14.5Hz), 5.10 (1H, d, J=14.5HZ), 5.29 (1H, d, J=4.3Hz), 5.83 (1H, d, J=4.3Hz), 7.99 (lH, s) Preparation 57 30 To a suspension of tert-butyl 3- aminopropylcarbamate (366 mg) and 3-ethoxy-4-{1-methy1- 5- (tritylamino) —1H—pyrazol-4-yl] amino)-3-cyclobutene— l,2-dione (670 mg) in ethanol (30 ml) was added triethylamine (0.195 ml), and the mixture was stirred 35 under reflux for 3 hours, To the reaction mixture were added diethyl ether (40 ml) and hexane (10 ml). The crystalline precipitate was collected by filtration and dried in vacuo to give tert-butyl (3-[(2-[(l-methyl-5- 81 WO 2004/039814 PCT/JP2003/013684 (tritylamino) -lH-pyrazol-4-yl] amino }-3 ,4-dioxo-l- cyclobuten- 1-yl) amino] propyl} carbamate (783 mg) as a solid. 1H-NMR(CDC13) 1.43 (9H, s), 1.67 (2H, quintet, J=5.5Hz), 5 3.15 (2H, q, J=5.5Hz), 3.17 (3H, s) , 3.60 (2H, q, J=5.5Hz), 4.82 (1H, brs), 4.86 (1H, t, J=5.5Hz), 5.44 (1H, br), 5.86 (1H, br), 7.13-7.33 (15H, m), 7.17 (1H, s) Example 33 10 To a solution of benzhydryl 7-[(Z)-2-(5-amino- 1,2, 4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1- methylethoxyimino) acetamido] -3-iodomethyl-3—cephem-4— carboxylate (819 mg) in N,N-dimethylformamide (1.6 ml) was added N-trimethylsilyl) acetamide (656 mg) , and the 15 mixture was stirred at room temperature for 40 minutes. To the reaction mixture was added a solution of tert- butyl (3-[(2-{[l-methyl-5-(tritylamino)-lH-pyrazol-4- yl]amino}-3,4-dioxo-l-cyclobuten-l- yl) amino]propyl)carbamate (637 mg) in N,N- 20 dimethylformamide (3.2 ml) , and the whole mixture was stirrd at 35-40°C for 3.5 hours. To the resulting reaction mixture was added ethyl acetate (60 ml) and the precipitate was filtered off. The filtrate was washed successively with water (50 ml x 2) and brine (50 ml), 25 dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to about 8 ml in vacuo. The concentrate was poured into diisopropyl ether (80 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the 30 resulting solid in methylene chloride (2.4 ml) were added anisole (0.80 ml) and trifluoroacetic acid (1.6 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether (80 ml). The resulting precipitate was collected 35 by filtration and dried in vacuo to give a crude product (565 mg) , which was purified by preparative HPLC utilizing ODS column eluting with a mixture of acetonitrile and phosphate buffer (pH 5.5). The eluate 82 WO 2004/039814 PCT/JP2003/013684 containing a desired product was concentrated to about 20 ml in vacuo. The concentrate was desalted by prepative HPLC utilizing ODS column ,and the fraction eluted with 8 % acetonitrile/0.01 m hydrochloric acid was 5 concentrated to about 10 ml in vacuo and lyophilized to give 3-([3- amino-4-[(2-[(3-aminopropyl ) amino]-3,4-dioxo 1-cyclobuten-l-yl} amino) -2-methyl-l-pyrazolio] methyl} - 7 b -[ (Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(l-carboxy- 1-methylethoxyimino) acetamido) -3-cephem-4-carboxylate 10 trihydrochloride (34 mg) as an amorphous solid. 1H-NMR(D2O) 1.62 (6H, s), 2.02 (2H, quinter, J=7.3Hz), 3.09 (2H, t, J=7.3HZ), 3.32 (1H, d, J=17.5Hz), 3.54-3.65 (lH, m), 3.67-3.78 (2H, m) , 3.75 (3H, s), 4.93-5.23 (2H, m), 5.30 (1H, d, J=4.5Hz), 5.36 (1H, d, J=4.5Hz), 7.99 l5 (1H, s) Preparation 58 To a solution of 1,1-(1, 2-dioxo-l, 2- ethanediyl)bis-lH-imidazole (761 mg) in N,N- dimethylformamide (8 ml) was added l-methyl-N5-trityl- 20 lH-pyrazole-4,5-diamine (709 mg) under ice-cooling, and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added a solution of tert-butyl 2-aminoethylcarbamate (1.28 g) in N,N- dimethylformamide (2 ml) , and the mixture was stirred at 25 room temperature for 27 hours. To the reaction mixture was added ethyl acetate (50 ml) . After the precipitate was filtered off, the filtrate was washed successively with water, 5% aqueous citric acid solution and brine. The organic layer was dried over anhydrous sodium 30 sulfate, filtered and concentrated in vacuo. The crystalline residue was washed with a mixed solvent of diethyl ether and ethyl acetate and dried in vacuo to give tert-butyl {2-[ (2-{[l-methyl-5-(tritylamino)-1H- pyrazol-4-yl]amino}-2-oxoacetyl]amino}ethyl)carbamate 35 (823 mg) as a solid. 1H-NMR(CDC13) 1.43 (9H, s) , 2.97 (3H, s) . 3.31 (2H q, J=5.5Hz), 3.43 (2H, q, J=5.5HZ), 4.53 (1H, s), 4.84 (1H, brs) , 7. 10-7. 30 (15H, m) , 7.47 (1H, s) , 7.67 (1H, brs) , 83 WO 2004/039814 PCT/JP2003/013684 8.20 (1H, brs) Example 34 To a solution of 4-methoxybenzyl 7-[(Z)-2—{5- amino-1, 2 , 4-thiadiazol-3-Yl) -2- (l-tert-butoxycarbonyl-l- 5 methylethoxyimino) acetamido] - 3-chloromethyl-3-cephem-4- carboxylate (618 mg) in N,N-dimethylformamide (l.5 ml) was added N- (trimethylsilyl) acetamide (656 mg) , and the mixture was stirred at room temperature for 40 minutes. To the solution waa added potassium iodide (232 mg), and 10 the mixture was stirred at room temperature for 35 minutes. TO the reaction mixture was added a solution of tert-butyl (2-[(2-{[l-methyl-5-(tritylamino)-1H- pyrazol-4—yl] amino]-2-oxoacetyl)amino]ethyl]carbamate (626 mg) in N,N—dimethylformamide (3 ml), and the whole 15 mixture was stirred at 35-40°C for 24 hours. To the resulting reaction mixture was added ethyl acetate (50 ml) , and the solution was washed successively with water (50 ml x 2), 10% aqueous sodium trifluoroacetate solution (50 ml x 2) and brine (50 ml), dried over 20 anhydrous sodium sulfate and filtered. The filtrate was concentrated to about 10 ml in vacuo. The concentrate was poured into diisopropyl ether (60 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the solid in methylene 25 chloride (2.9 ml) were added anisole (0.95 ml) and trifluoroacetic acid (2.9 ml). The resulting solution was stirred at room temperature for 4 hours and poured into diisopropyl ether (60 ml) . The resulting precipitate was collected by filtration and dried in 30 vacuo to give a crude product (770 mg), 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 with concentrated hydrochloric acid and 35 chromatographad on Diaion® 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 3-{[3-amino-4-({2-[(2- 84 WO 2004/039814 PCT/JP2003/013684 aminoethyl) amino] -2-oxoacetyl} aininio) -2-methyl-l- pyrazolio]methyl] -7 b - [ (Z) -2-(5-amino-l , 2 , 4-thiadiazol-3- yl)-2-(1-carboxy-l-methylethoxyimino)acetamido]-3- cephem-4-carboxylate (31 mg) as an amorphous solid. 5 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s), 3.20 (1H, d, J=l8.0Hz), 3.24 (2H, t, J=6.0Hz), 3.45 (1H, dr J=18.0Hz ) 3.66 (2H, t, J=6.0Hz), 3.75 (3H, s) , 5.02 (1H, d, J=15.5Hz), 5,21 (1H, d, J=l5.5Hz), 5.25 (1H, d, J=5.0Hz), 5.85 (1H, d, J=5.0Hz), 8.14 (1H, s) 10 Preparaaation 59 To a suspension of phenyl {l-methyl-5- tert-butyl 3-azetidinylcarbamate acetic acid salt (418 mg) in methylene chloride (8 ml) was added N- 15 ethyldiisopropylamine (0.62 ml) , and the mixture was stitred under reflux for 16 hours. To the reaction mixture was added methylene chloride, and the solution was washed successively with 10% aqueous citric acid solution, 10% aqueous sodium hydroxide solution and 20 brine. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with a mixed solvent of ethyl acetate and hexane to give tert-butyl [1-([[1-methyl-5- (tritylamino)-lH-pyrazol-4—yl]amino ) carbonyl)-3 — 25 azetidinyl]carbamate (735 mg) as a solid, 1H-NMR(CDCl3) 1.47 (9H, s) , 2.92 (3H, s) , 3.56 (2H, dd, J=7.5, 5.0Hz), 4.02 (2H, dd, J=7.5, 7,5HZ), 4.42 (lH, brs), 4.71 (lH,s), 4.74 (1H, s), 4.94 (1H, brs), 7.18- 7.21 (7H,m) , 7.25-7.32 (9H, m) 30 Example 35 To a solution of benzhydryl 7-[ (Z)-2-(5-amino- l,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-l- methylethoxyimino) acetamido ] -3-iodomethyl-3-cephem-4- carboxylate (819 mg) in N,N-dimethylformamide (2.4 ml) 35 was added N-(trimethylsilyl) acetamide (655 mg) , and the mixture was stirred at room temperature for 3 0 minutes. To the reaction mixture was added a solution of tert- butyl [1-({[1-methyl-5-(tritylamino)-lH-pyrazol-4- 85 WO 2004/039814 PCT/JP2003/013684 yl] amino} carbonyl]— 3-acetidinyl] carbamate (553 mg) in N,N-dimethylformamide (3 ml) , and the whole mixture was stirred at room temperature for 3 hours, and then stirred at 50°C for 1 hour. To the resulting reaction 5 mixture were added ethyl acetate (50 ml) and water 50 ml) . The aqueous layer was separated, and the organic layer was washed with water and brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The 10 concentrate was poured into diisopropyl ether (80 ml) , and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene chloride (2.1 ml) were added anisole (0.7 ml) and trifluoroacetic acid (2.1 ml), 15 The resulting solution was stirred at room temperature for 4.5 hours and poured into diisopropyl ether (80 ml). The resulting precipitate was collected by filtration and dried in vacuo to give a crude product (521 mg) , which was purified by preparative HPLC utilizing ODS 20 column eluting with a mixture of acetonitrile and phosphate buffer (pH 5.5). The eluate containing a desired product was concentrated to about 20 ml in vacuo The concentrate was desalted by preparative HPLC utilizing ODS column, and the fraction eluted with 7% 25 acetonitrile/0.01 M hydrochloric acid was concentrated to about 10 ml in vacuo and lyophilized to give 3—[(3— amino-4-{ [ (3-amino-l-azetidinyl) carbonyl] amino] -2- methyl-1-pyrazolio) methyl]-7-[ (Z) -2- (5-amino-l,2,4- thiadiazol-3-yl)-2-(1-carboxy-l- 30 methylethoxyimino) acetamido]—3-cephem-4-carboxylate trihydrochloride (22 mg) as an amorphous solid. 1H-NMR(D2O) 1.62 (3H, s), 1.63 (3H, s), 3.25 (lH, d, J=17.9Hz) , 3.50 (1H, d, J=17.3Hz), 3.72 (3H, s) , 4.14 (2H, dd, J=9.6, 4.4Hz), 4.25 (1H, tt, J=7.8, 4.6Hz), 35 4.46 (2H, dd, J=9.6, 7.8Hz), 5.08 (1H, d, J=15.6HZ), 5.24 (1H, d, J=15.6Hz), 5.27 (1H, d, J=4.6Hz), 5.88 (1H, d, J=4.6HZ), 7.91 (lH, s) Preparation 60 86 WO 2004/039814 PCT/JP2003/013684 To a suspension of phenyl [l-methyl-5- (tritylamino) -lH-pyrazol-4-yl] carbamate (2. 18 g) and tert-butyl 3-amino-l-azetidinecarboxylate (7 93 mg) in methylene chloride (30 ml) was added N- 5 ethyldiisopropylamine (1.07 ml), and the mixture was stirred under reflux for 40 hours. To the reaction mixture was added methylene chloride, and the solution was washed successively with 10% aqueous citric acid solution, 10% aqueous sodium hydroxide solution and 10 brine. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatogrsphy on silica gel eluting with 10% methanol/methylene chloride to give tert-butyl 3- [{ (l-methyl-5- (tritylamino) -1H- 15 pyrazol-4—yl]amino)carbonyl)amino]-1- acetidinecarbonylate (1.52 g) as a solid. 1H-NMR(CDCl3) 1.44 (9H, s) , 3.03 (3H, s), 3.59 (2H, dd, J=9.2, 5.0Hz), 4.17 (2H, dd, J=9.2, 7.8Hz), 4.39-4.43 (3H, m) , 4.64 (lH, brs), 7.19-7.21 (1H, m) , 7.27 (1H,s), 20 7.29-7.32 (9H, m) Example 36 To a solution of 4-methoxybenzyl 7-[(Z)-2-(5- amino-1,2,4-thiadiazol-3-yl)-2-(l-tert-butoxycarbonyl-l- methylethoxyimino)acetamido]—3-chloromethyl-3-cephem-4- 25 carboxylate (1.48 g) in H,N-dimethylformaipide (3.0 ml) was added N- (trimethylsilyl) acetamide (1.42 g) , and the mixture was stirred at room temparature for 30 minutes. TO the solution was added potassium iodide (504 mg) and the mixture was stirred at room temperature for 30 30 minutes. To the reaction mixture was added a solution of tert-butyl 3-[({1-methyl-5-(tritylamino)-1H-pyrazol- 4—yl] amino } carbonyl) amino) -1-azetidinecarboxylate (1.20 g) in N,N-dimethylformamide (2.2 ml), and the whole mixture was stirred at 50°C for 16 Hours, To the 35 resulting reaction mixture was added ethyl acetate (200 ml), and the solution was washed successively with water (50 ml), 10% aqueous sodium trifluoroacetate solution (50 ml x 2) and brine (50 ml) , dried over anhydro-us 87 WO 2004/039814 PCT/JP2003/013684 sodium sulfate and filtered. The filtrate was concentrated to about 10 ml in vacuo. The concentrate was poured into diisopropyl ether (160 ml) , and tne resulting precipitate was collected by filtration and 5 dried in vacuo. To a solution of the solid in methylene chloride (8.64 ml) were added anisole (2.88 ml) and frifluoroacetic acid (8.64 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether (160 ml). The resulting 10 precipitate was collected by filtration and dried in vacuo to give a crude product (2.22 g), which was purified by preparative HPLC utilizing ODS column eluting with, a mixture of acetonitrile and phosphate buffer (pH 5.5). The eluate containing a desired 15 product was concentrated to about 20 ml in vacuo. The concentrate was desalted by preparative HPLC utilizing ODS column, and the fraction eluted with 8% aqueous acetonitrile was concentrated to about 10 ml in vacuo and lyophilized to give 3-[(3-amino-4-{[(3- 20 azetidinylamino)carbonyl)amino}—2-methy1-1- pyrazolio)methyl]-7-2-(5-amino-l,2,4-thiadiazol-3- yl)-1— (1—carboxy—1-methylethoxyimino)acetamido]-3- cephem—4—carboxylate (220 mg) as an amorphous solid. 1H-NMR(D2O) 1.50 (3H, s), 1.51 (3H, s), 3.20 (1H, d, 25 J=17.6Hz), 3.47 (lH, d, J=17.6Hz), 3.70 (3H, s), 4,18 (2H, dd, J=11.2, 7.6Hz), 4.31 (2H, dd, J=11.2, 8.3Hz), 4.68 (1H, tt, J=8.3, 7.6HZ), 4.94 (1H, d, J=l5.6Hz), 5.15 (1H, d, J=15.6HZ), 5.23 (1H, d, J=4.8Hz), 5.83 (1H, d, J=4.8Hz), 7.87 (1H, s) 30 Preparation 61 To a suspension of phenyl [l-methyl-5- (tritylamino)-lH-pyrazol-4-yl]carbamate (786 mg) and tert-butyl 3-pyrrolidinylcarbamate (373 mg) in methylene chloride (6 ml) was added N-ethyldiisopropylamine (0.43 35 ml), and the mixture was stirred under reflux for 10 hours. The reaction mixture was washed successively with 10% aqueous citric acid solution, brine and saturated aqueous sodium hydrogen carbonate solution. 88 WO 2004/039814 PCT/JP2003/013684 The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give tert-butyl {l-({[l-methyl-5- (tritylamino) -lH-pyrazol-4- yl] amino)carbonyl) -3-pyrrolidinyl] carbamate (730 mg) as 5 a solid. 1H-NMR(CDCl3) 1.43 (9H, s), 1.82-1.88 (1H, m) , 2.12- 2.18 (1H, m), 2.39 (3H, s), 2.89-3.03 (1H, m), 3.20-3.30 (2H, m) , 3.33-3.43 (1H, m) , 4.22 (1H, br), 4.69 (1H, br), 4.88 (1H, brs), 4.96 (1H, brs), 7.18-7.27 (16H, m) 10 Example 37 To a solution of benzhydryl 7[(Z)-2-(5-amino- l,2,4-thiadiazol-3-yl) -2- (1-tert-butoxycarbonyl-l- methylethoxyimino)acetamido]-3-iodomethyl-3—cephem—4— carboxylate (819 mg) in N,N-dimethylformamide (2.4 ml) 15 was added N- (trimethylsilyl) acetamide (655 mg) , and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added a solution, of tert- butyl [1- ({[1-methyl-5- (tritylamino)-lH-pyrazol-4- yl] amino} carbonyl) -3—pyrrolidinyl] carbamate (567 mg) in 20 N,N-dimethylformamide (3.0 ml) . The whole mixture was stirred at room temperature for 3 hoors. To the resulting reaction mixture were added ethyl acetate (100 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed successively with 10% 25 aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 2.5 ml in vacuo. The concentrate was poured into diisopropyl ether (80 ml), and the resulting precipitate 30 was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene chloride (2.55 ml) were added anisole (0.85 ml) and trifluoroacetic acid (2.55 ml), and the mixture was stirred at room temperature for 3 hours. The reaction 35 mixture was poured into diisopropyl ether (80 ml), and the resulting precipitate was collected by filtration, and dried in vacuo to give a crude product (6 08 mg) , which was purified by preparative HPLC utilizing ODS 89 WO 2004/039814 PCT/JP2003/013684 column eluting with a mixture of acetonitrile and phosphate buffer (pH 5.5), The eluate containing a desired product was concentrated to about 20 ml in vacua. The concentrated was desalted by preparative HPLC 5 utilizing ODS column, and the fraction eluted with 7% acetonitrile/0.01 M hydrochloric acid was concentrated to about 10 ml in vacuo and lyophilized to give 3- [ (3- amino-4- { [( 3-amino-l-pyrrolidinyl) carbonyl ) amino} -2- methyl-l-pyrazolio)methyl]-7-[(Z)-2-(5-amino-l,2,4- 10 thiadiazol-3-yl) -2- {1 -carboxy-1 - methylethoxyimino)acetamido]-3-cephem-4-carboxylate trihydrochloride (31 mg) as an amorphous solid. 1H-NMR(D20) 1.61 (3H, s), 1.61 (3H, s), 2.13-2.27 (1H, m) , 2.39-2.54 (1H, m) , 3.25 (1H, d, J=l8.lHz) , 3.51 (1H, 15 d, J=18.1HZ), 3.55-3.68 (3H,m), 3.73 (3H, s), 3.80 (1H, dd, j=ll.5, 6.0Hz), 4.01-4.11 (lH, m), 5.20 (1H, d, J=16.0Hz), 5.24 (1H, d, J=16.0Hz), 5.29 (1H, d, J=4.8Hz), 5.89 (1H, d, J=4.8Hz), 7.91 (1H, s) Preparation 62 20 To a suspension of phenyl [ 1-methy 1-5- (tritylamino)-lH-pyrazol-4-yl]carbamate (711 mg) and tert-butyl 3-amino-l-pyrrolidinecarboxylate (372 mg) in methylene chloride (15 ml) was added N- ethyldiisopropylamine (0.51 ml) , and the mixture was 25 stirred under reflux for 17 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 30 in vacuo, The residue was purified by column hromatography on silica gel eluting with 10% ethanol/methylene chloride to give tert-butyl 3-[{(l- ethy 1-5- (tritylamino) -lH-pyrazol-4- 1]amino ) carbonyl) amino]—l-pyrrolidinecarboxylate (511 35 mg) as a solid. 1H-NMR(CDCl3) 1.46 (9H, s) , 1.66-1.74 (1H, m) , 2.04- 2.11 (1H, m) , 2.97 (3H, s), 3.05-3.11 (lH, m) , 3.30-3.43 (2H, m) , 3.53-3.58 (1H, m), 4.16-4.23 (2H, m), 4.45 {1H, 90 WO 2004/039814 PCT/JP2003/013684 brs), 4.74 (1H, br), 7.18-7.20 (6H, m), 7.28-7.30 (10H, m) Example 3 8 To a solution of benzhydryl 7- [(Z) -2- (5-amino- 5 1,2 ,4-thiadiazol-3—yl) —2 — (1-tert-butoxycarbonyl—1- methylethoxyimino) acetamido] — 3— iodomethyl-3—cephem-4- carboxylate (707 mg) in N,N-dimethylformamide (2.1 ml) was added N-(trimethylsilyl)acetamide (566 mg), and the mixture was stirred at room temperature for 30 minutes. 10 To the reaction mixture was added a solution of tert- butyl 3- [ { { [l-methyl-5- (tritylamino) -lH-pyrazol-4- yl]amino) carbonyl)amino] -1-pyrrolidinecarboxylate (490 mg) in N,N-dimethylformamide (2.0 ml) , The whole mixture was stirred at room temperature for 3 hours. To 15 the resulting reaction mixture were added ethyl acetate (100 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed successively with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried 20 over sodium sulfate and filtered. The filtrate was concentrated to about 3 ml in vacuo. The concentrate was poured into diisoprbpyl ether (80 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in 25 methylene chloride (1.83 ml) were added aniaole (0.61 ml) and trifluoroacetic acid (1.83 ml), and the mixture was stirred at room temperature for 5 hours. The reaction mixture was poured into diisopropyl ether (80 ml), and the resulting precipitate was collected by 30 filtration and dried in vacuo to give a crude product (440 mg), which was purified by preparative HPLC utilizing ODS column. The eluate containing desired products was concentrated to about 30 ml in vacuo. The concentrate was adjusted to about pH 3 witli concentrated 35 hydrochloric acid and chromatographed on Diaion® 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-[(3- 91 WO 2004/039814 PCT/JP2003/013684 amino-2-methyl-4- {[ (3-pyrrolidinylamino) carbonyl] ami no} - 1-pyrazolio) methyl] -7- [ (Z) -2- (5-amino-l,2, 4-thiadiazol- 3-yl] -2-(1-carboxy-l-methylethoxyimino) acetamido] -3- cephem-4-carboxylate (18 mg) as an amorphous solid. 5 1H-NMR(D2O) 1.54 (3H, s) , 1.55 (3H, s), 2.00-2.10 (lH, m) , 2.30-2.40 (1H, m) , 3.23 (0.5H, d, J=17.9Hz), 3.24 (0.5H, d, j=l7.9Hz), 3.27-3.34 (1H, m), 3.34-3.43 (1H, m) , 3.45-3.57 (3H, m) , 3.7Z (3H, s) , 4.36-4.46 (1H, m) , 4.95 [0.5H, d, J=l5.lHz), 4.96 (0.5H, d, J=15.6Hz), 5.17 10 (1H, d, J=15. 6Hz), 5.26 (1H, d, J=5. OHz) , 5.85 (1H, d, J=5.0Hz), 7.88 (1H, s) Preparation 6 3 To a suspension of tert-butyl {2- [ ( ( [ 1-methyl-5- (tritylamino)-lH-pyrazol-4-yl]amino}carbonyl)amino]- 15 ethyl]carbaramate (10.8 g) in methanol (50 ml) was added 4M hydrogen chloride solution in dioxane (50 ml) . The mixture was stirred at room temperature for 3 hours. The solvent was concentrated in vacuo, and the residue was triturated with ethyl acetate and dried in vacuo to 30 give N-(2-aminoethyl)-N'-(5-amino-l-methyl-1H-pyrazo1-4- yl)urea trihydrochloride (5.6 g) as a solid. 1H-NMR(DMSO-d6) 2.84-2.87 (2H, m) , 3.30 (2H, brs), 3.71 (3H, 3), 6.57 (lH, br), 7.91 (1H, s), 3.05 (4H,br), 8.55 (1H, br) 25 Preparation 64 To a solution of N-(2-aminoethyl)-N ' - (5—amino-1- methyl-1H-pyrazol-4-yl) urea trihydrochloride (3.1 g) and triethylamine (4.6 g) in chloroform (100 ml) was added di-tert-butyl [{ (trifluoromethyl] sulfonyl] imino)- 30 methylene) biscarbamate (5.9 g) . The mixture was stirred at room temperature for 90 minutes. The reaction mixture was washed successively with 10% aqueous citric acid solution, brine and saturated, aqueous sodium hydrogen carbonate solution. The organic layer was 35 dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with ethyl acetate to give di-tert-butyl[ (Z)-{ [2-{{{5- amino-l-methyl-lH-pyrazol-4-yl) amino] carbonyl} amino} - 92 WO 2004/039814 PCT/JP2003/013684 ethy1]amino}methylidene)bis carbamate (4.3 g) as a solid. 1H-NMR(DMSO-d6) 1.39 (9H, s) , 1.48 (9H, s) , 3.18 (2H, q, J=6.0HZ), 3.35 (2H, br), 3.49 (3H, s), 4.77 (1H, brs), 6.05 (1H, br) , 6.97 (1H, s), 7,.9 (1H, brs) , 8.36 (1H, t, 5 J=5.5Hz), 11.49 (1H, brs) Preparation _65 j To a solution of di-tert-butyl { (Z)-{ [2-({[(5- amino-1-methyl-1H—pyrsazol-4-yl) amino] carbonyl ]amino)- ethyl] amino}methylidene) bis carbamate (2.2 g) and 10 triethylamine (0.6 g) in chloroform (3 0 ml) was added trityl chloride (1.7 g) , and the mixture was stirred at room temperature for 14 hours. The reaction mixture was washed successively with 10% aqueous citric acid solution, brine and saturated aqueous sodium hydrogen 15 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 di-tert-butyl [ (Z) - [ {2- [ ({ [l-methyl-5- (tritylamino)-lH-pyrazol-4-yl] amino] carbonyL} amino]- 20 ethyl}amino}methylidene]biscarbamate (1.9 g) as a solid. 1H-NMR(DMSO-d6) 1.39 (9H, s), 1.47 (9H, s), 2.72 (3H, s), 3.09-3.10 (2H, m), 3.31-3.34 (2H, m), 5.69 (1H, s), 6.10 (1H, br), 6.77 (1H, brs) , 7.02 (1H, s) , 7. 14-7.16 (6H, m) , 7.22-7.27 (9H, m), 3.36 (lH, t, J=5.5Hz), 11.51. 35 (1H, brs) Example 39 T:o a solution of benzhydryl 7 [ (Z) -2- (5—amino— l,2,4-thiadiazol-3-yl) -2- [l-tert-butoxycarbonyl-l- methylethoxyimino) acetamido) -3 - iodomethy 1-3-cephem-4- 30 carboxylate (820 mg) in N,N-dimethylformamide (1.4 ml) was added N-(trimethylsilyl) acetamide (6 56 mg), and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture were added di-tert-butyl [(Z)- ((2- [ ( { [1-methyl-5- (tritylamino) -lH-pyrazol-4- 35 yl ] amino} carbony 1) amino ] ethyl} amino) methylidane ] - biscarbamate (820 mg) and N,N--dimethylformamide (2.0 ml). The whole mixture was stirred at room temperature for 3 hours . To the resulting reaction mixture were added 93 WO 2004/039814 PCT/JP2003/013684 ethyl acetate (100 ml) and water (50 ml). The aqueous layer was separated, and the organic layer was washed successively with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and 5 brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuci. The concentrate was poured into diisopropyl ether (120 ml) , and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the 10 resulting solid 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 4 hours. The reaction mixture was poured into diisopropyl ether (100 ml) , and the resulting precipitate was 15 collected by filtration and dried in vacuo to give a crude product (740 mg), 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 20 pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® 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-{[3-amino-4-({[2- 25 guanidinoethyl)amino] carbonyl) amino) -2-methyl—1 — pyrazolio [methyl]-7- [ (Z) -2- (5-amino-l, 2 , 4-thiadiazol-3- yl) -2—(1-carboxy—1—methylethoxyimino)acetamido]-3— cephem-4-carboxylate (70 mg) as an amorphous solid. 1H-NMR(D2O) 1.55 (3H, s), 1.56 (3H, s), 3.24 (1H, d, 30 J=17.6HZ), 3.28-3.40 (4H, m), 3.52 (lH, d, J=17.5HZ), 3.73 (3H, s), 4.97 (1H, d, J=15.4Hz), 5.16 (1H, d, J=15.4Hz), 5.27 (1H, d, J=4.8Hz) , 5.8 4 (1H, d, J=4.8Hz), 7.37 (1H, s) Preparation 66 35 To a suspension of phenyl [1—methyl-5- (tritylamino)-lH-pyrazol-4-yl]carbamate (9 50 mg) and tert-butyl (3S)-3-pyrrolidinylcarbamate (560 mg) in methylene chloride (20 ml) was added, N- 94 WO 2004/039814 PCT/JP2003/013684 ethyldiisopropylamine (1350 mg), and the mixture was stirred under reflux for 23 hours. The reaction mixture was washed successively with 10% aqueous citric acid solution, brine and saturated aqueous sodium hydrogen 5 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 4% methanol/chloroform to give tert-butyl [ (3S) -1- { { [1- 10 methyl-5- (tritylamina) -1H-pyrazol—4-yl] amino] carbonyl)-. 3—pyrrolidinyll carbamate (680 mg) as a solid. 1H-NMR(CDCl3) 1.48 (9H,s) , 1.82-1.86 (1H, m) , 2.12- 2.18 (1H, m) , 2.89 (3H, s), 2.89-3.03 (1H, m), 3.20-3.30 (2Hr in), 3.3B-3.43 (IE, m) r 4 .22 (lHr br) t 4.69 [1M, bri) , 15 4.88 (lH, brs), 4.96 (1H, brs), 7.18-7.27 (16H, m) Example 40 To a solution of benzhydryl 7 [ (Z) -2- (5-amino- 1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1- methylethoxyimino) acetamido] -3-iodomethyl—3-cephem-4- 20 carboxylate (320 mg) in N,N-dimethylformamide (2.4 ml) was added N-(trimethylsilyl)acetamide (656 mg), and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added tert-butyl [(3S)-1- ({[l-methyl-5-(tritylamino)-lH-pyrazol-4- 25 yl] amino}carbonyl)-3—pyrrolidinyl}-carbamate (680 mg). The whole mixture was stirred at room temperature for 3 hours. To the resulting reaction mixture were added ethyl acetate (80 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed 30 successively with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (120 ml), 33 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 , 95 WO 2004/039814 PCT/JP2003/013684 and the mixture was stirred at room temperature for 4 hours. The reaction mixture was poured into diisopropyl ether (10 0 ml), and the resulting precipitate was collected by filtration and dried in vacuo to give a 5 crude product (690 mg) , 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 with concentrated hydrochloric acid and 10 chromatographed on Diaion® 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-{{3-amino—4-({[(3S)-3—amino-1- pyrrolidinyl]carbonyl]amino)-2—methyl-1— 15 pyr azolio] methyl}-7 - [(Z)-2-95-amino-l, 2 , 4-thiadiazol-3- yl) -2- (l-carboxy-l-methylethoxyimino) acetamido] -3- cephem-4-carboxylate (60 mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (6H, s), 2.13-2.27 (1H, m), 2.38-2.53 (1H, m), 3.20 (1H, d, J=17.4Hz), 3.46 (1H, d, J=17.4Hz), 20 3.54-3.67 (3H, m) , 3.73 (3H, s), 3.79 (1H, dd, J=11.5, 6.0Hz), 4.00-4.10 (1H, m), 4.97 (1H, d, J=15.4Hz), 5.16 (1H, d, J=15.4Hz), 5.25 (lH, d, J=4.8Hz), 5.83 (1H, d, J=4.8Hz), 7.85(1H, s) Preparation 67 25 To a suspension of phenyl [ 1-methyl-5- (tritylamino)-1H—pyrazol-4-yl]carbamate (950 mg) and tert-butyl (3R)-3-pyrrolidinylcarbamate (5 60 mg) in methylene chloride (20 ml) was added N— ethyldiisopropylamine (39 0 mg), add the mixture was 30 stirred under reflux for 23 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 35 in vacuo. The residue was purified by column chromatography on silica gel eluting with 4% methanol/chloroform to give tert-butyl [(3R)-1-{{[1- methyl-5- (tritylamino) -lH-pyrazol-4-yl] amino ) carbonyl ) - 96 WO 2004/039814 PCT/JP2003/013684 3-pyrrolidinyl]carbamate (700 mg) as a solid. 1H-HMR(CDC13) 1.48 (9H, s), 1.82-1.88 (1H, m) , 2.12- 2.18 (1H, m), 2.39 (3H, s), 2.89-3.03 (lH, m) , 3.20-3.30 (2H, m) , 3.38-3.43 (1H, m) , 4.22 (1H, br) , 4.69 (lH, br) , 5 4.38 (1H, brs), 4.96 (1H, brs), 7.18-7.27 (16H, m) Example 41 To a solution of benzhydryl 7-[ (Z)-2-(5-amino- 1,2, 4-thiadiazol-3-yl) -2- (1-tert-butoxycarbonyl-l- methylethoxyimino) acetamido] -3-iodomethyl-3-cephem— 4- 10 carboxylate (820 mg) in N,N-dimethylformamide (2.4 ml) was added N-(trimethylsilyl)acetamide (656 mg), and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added tert-butyl [(3R)—1— ({[1—methyl-5—(tritylamino)-lH-pyrazo1-4— 15 yl] amino}carbonyl}-3-pyrrolidinyl] carbamate (680 mg) . The whole mixture was stirred at room temperature for 3 hours. To the resulting reaction mixture were added ethyl acetate (80 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed 20 successively with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (120 ml), 25 and the is suiting precipitate was collacted 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) , and the mixture was stirred at room temperature for 4 30 hours. The reaction mixture was poured into diisopropyl ether (100 ml), and the resultlug precipitate was collected by filtration and dried in vacuo to give a crude product (7 60 mg) , which was purified by preparative HPLC utilizing ODS column. The eluate 35 contaiuing a desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to about pH 3 with concentrated hydrochloride acid and chromatographed on Diaion.® HP-20 (Mitsubishi Chemical 97 WO 2004/039814 PCT/JP2003/013684 Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 3 0 ml in vacuo and lyophilized to give 3-{[3-amino-4-{[(3R)-3-amino-l- pyrrolidinyl] carbonyl}amino) -2-methyl-1- 5 pyrazolio]methyl}-7-[(Z)-2-[5-amino-l,2,4-thiadiazol-3- yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3- (cephem-4-carboxylate (68 mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (6H, s) , 2.13-2.27 (1H, m) , 2.38-2.53 (1H, m) , 3.20 (1H, d, J=17.6Hz), 3.47 (1H, d, J=17.6Hz), 10 3.56-3.66 (3H, m) , 3.73 (3H, s), 3.79 (1H, dd, J=ll.0, 6.0Hz) , 4.00-4.10 (1H, m), 4.96 (1H, d, J=15.1Hz), 5.15 (1H, d, J=15.1Hz), 5.26 (1H, d, J=4.8Hz), 5.83 (1H, d, J=4.8Hz), 7.84 (1H, s) Preparation 68 15 To a suspension of phenyl (5-amino-l-methyl-lH- pyrazol-4-yl) carbamate (1.86 g) and (3s) -1-benzyl-3 - pyrrolidinamine (2.0 g) in chloroform (50 ml) was added N-ethyldiisopropylamine (3.1 g) , and the mixture was stirred under reflux for 19 hours. The reaction mixture 20 was concentrated in vacuo to give crude (S)—5-amino—4— [3-(l-benzyl-3-pyrrolidinyl)ureidol-l-methyl-lH-pyrazole as a solid. A solution of the crude product in acetic acid was treated with palladium black (3 ml) under a hydrogen atomosphere at room temperature for 24 hours. 25 After the catalyst was filtered off, the filtrate was concentrated in vacuo, and the residue was dissolved in saturated aqueous sodium hydrogen carbonate solution (100 ml) . To the solution was added a solution of di- tert-butyl dicarbonate (5.0 g) in tetrahydrofuran (40 30 ml} , and the mixture was stirred at room temperature for 5 hours. The reaction mixture was extracted with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diethyl ether to give 35 tert-butyl (3S)-3-({ [ (5-amino-l-methyl-lH-pyrazol-4- yl) amino] carbonyl]amino] -1-pyrrolidinecarboxylate (1.9 g) as a solid. 1H-NMR(DMSO-d6) 1.40 (9H, s), 1.70-1.76 (1H,m) , 1.95- 98 WO 2004/039814 PCT/JP2003/013684 2.02 (1H, m) , 3.01-3.05 (1H, m) , 3.24-3.34 (2H, m) , 3.38-3.45 (1H, m) 3.50 (3H, s), 4.06-4.11 (lH, m), 4.78 (2H, brs), 6.19 (1H, brs) , 6.97 (1H, s) , 7.09 (1H, brs) Preparation 69 5 To a solution of tert-butyl (3S) -3- {( [ (5-amino-1- methyl-lH-pyrazol-4-yl)amino] carbonyl)amino}-1- pyrrolidinecarboxylate (1.8 g) and N- ethyldiisopropylamine (720 mg) in chloroform (50 ml) was added trityl chloride (1.6 g) , and the mixture was 10 stirred at room temperature for 28 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 15 concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with 3% methanol/chloroform to give tert-butyl (3S)-3-[({1- methyl-5- (tritylamino)-l-pyrazol-4- yl]amino)carbonyl)amino]-1—pyrrolidinecarboxylate (1.7 20 g) as a solid. 1H-NMR(CDCl3) 1.45 (9H, s), 1.66-1.74 (1H, m), 2.04- 2.11 (1H, m), 2.97 (3H, s), 3.05-3.11 (1H, m), 3.30-3.43 (2H, m), 3.53-3-58 (1H, m) 4.16-4.23 (2H, m) , 4.45 (1H, brs) , 4.74 (lH, br), 7.18-7.20 (6H, m) , 7.28-7.30 (10H, 25 m) Example 42 To a solution of benzhydryl 7- [ (Z) -2- (5-amino- 1,2, 4-thiadiazol-3-yl) -2- (1-tert-butoxycarbonyl-l- methylethoxyimino) acetamido ] -3-iodomethyl-3-cephem-4- 30 carboxylate (820 mg) in N,N-dimethylformamide (2.4 ml) was added N- (trimethylsilyl) acetamide (656 mg) , and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added tert-butyl (3S) -3- [ ({ [1-methyl-5- (tritylamino) -lH-pyrazol-4- 35 yl] amino} carbonyl) amino]-1-pyrrolidinecarboxylate (6 80 mg) . The whole mixture was stirred at room temperature for 3 hours. To the resulting reaction mixture were added ethyl acetate (80 ml) and water (50 ml) . The 99 WO 2004/039814 PCT/JP2003/013684 aqueous layer was separated, and the organic layer was washed successively with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried over sodium 5 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 a solution of the resulting solid in 10 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 5 hours. The reaction mixture was poured into diisopropyl ether (10 0 ml), and the resulting precipitate was collected by filtration 15 and dried in vacuo to give a crude product (87 0mg) , 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 with concentrated 20 hydrochloric acid and chromatographed on Diaion® 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-{[3- amino-2—methyl—4—({[(3s)-3-pyirrolidinylamino]carbonyl}- 25 amino} -1-pyrazolio]methyl)-7- [(Z) -2- (5-amino-l ,2,4- thiadiazol—3-yl)-2-(1-carboxy-1-methylethoxyimino)- acetamido]-3-cephem-4-carboxylate (6 8 mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s), 2.00-2.09 (1H, 30 m) , 2.28-2.38 (1H, m) , 3.22 (1H, d, J=17.4Hz), 3.29 (1H, dd, J=12.4, 4.6Hz), 3.34-3.42 (1H, m), 3.44-3.54 (3H, m), 3.71 (3H, s), 4.36-4.43 (1H, m), 4.95 (lH, d, J=15,6Hz), 5.15 (1H, d, J=15.6Hz), 5.25 (1H, d, J=4.6Hz), 5.84 (lH, d, J=4.6Hz), 7.87 (1H, s) 35 Preparation_7 0 To a suspension of 4-[(tert— butoxycarbonyl) amino] butanoic acid (2.13 g) in dichloromethana (40 ml) was added l-hydroxybenzotriazole. 100 WO 2004/039814 PCT/JP2003/013684 (HOBT) (1.41 g) and N- (3-dimethylaminopropyl)-N' - ethylcarbodiimide hydrochloride (WSCD HC1) (3.65 g) , and the mixture was stirred for 1 hour. To the solution were added 1-methyl-lH-pyrazole-4,5-diamine sulfate (2 5 g) and N, N—diisopropylethylamine (3.32 ml), The reaction mixture was stirred for 18 hours. To the resulting solution were added brine and saturated aqueous sodium hydrogen carbonate solution, and the mixture was extracted with ethyl acetate. The aqueous 10 layer was extracted with tetrahydrofuran/ethyl acetate= 1/1 twice. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. To the residue waS added pyridine (40 ml) , and then added chlorotriphenylmethane (5.3 g) , The mixture was 15 stirred at 65oC for 6 hours. The mixture was dissolved in ethyl acetate. Ths 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 20 residue was purified by column chromatography on silica gel eluting with 60% ethyl acetate/dichloronethane to give tert-butyl {4-{ [l-methyl-5- [tritylamiiao) -1H- pyrazol-4—yl] amino }-4—oxobutyl) carbamate (2.01 g) . 1H-NMR(CDCl3) 1.44 (9H, s) , 1.67 (2H, tt, J=6,7, 6.7Hz), 25 1.92 (2H, t, J=6.7Hz), 2.90 (3H, s), 3.09 (2H, dt, J=6.7, 6.7HZ), 4.50 (lH, s), 4.71 (1H, t, J=6.7Hz), 6.53 (1H, s) , 7.0-7.35 (16H, m) , 7.56 (1H, s) Example 43 To a solution of benzhydryl 7—[(Z) -2- (5-amino— 30 1,2 ,4-thiadiaEzl-3-yl)-2-(l-tert-butoxycarbonyl-l- methylethoxyimino) acetamido] -3-iodomethyl-3-cephem-4- carboxylate (2 g) in N,N-dimethylformamide (6 ml) was added N-(trimethylsilyl) acetamide (1.77 g) , and the mixture was stirred at room temperature for 30 minutes. 35 To the reaction mixture was added tert-butyl {4-{[1- methyl-5-{tritylamino)-lH-pyrazol-4-yl]amino}-4- oxobutyl)carbamate (1.93 g), and the whole mixture was stirred at 35oC for 30 hours. To the resulting reaction 101 WO 2004/039814 PCT/JP2003/013684 mixture was added ethyl acetate, and the solution was washed successively with water, 10% aqueous sodium trifluoroacetate solution and brine, dried over magnesium sulfate and filtered. The filtrate was 5 concentrated to about 25 ml in vacuo. The concentrate was poured into diisopropyl ether (150 ml) , and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the solid in methylene chloride (5 ml) were added anisole (1.5 ml) and 10 trifluoroacetic acid (5 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 (1.2 g). The crude product was dissolved 15 in a mixture of phosphate buffer (pH 6.86, 10 ml) and saturated aqueous sodium hydrogen carbonate solution and purified by preparative HPLC utilizing ODS column. The eluate containing a desired product was concentrated to about 20 ml in vacuo. The concentrate was adjusted to 20 about pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical Corporation) eluting with 20% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo, and 2M aqueous sulfuric acid solution (72 ml) was added. The 25 mixture was lyopliilized to give 3- { (3-amino-4-[(4- aminobutanoyl} amino] -2-methyl-l-pyrazolio}methyl) —7— [(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(1-carboxy-l- methylethoxyimino)acetamido]-3—cephem-4-carboxylic acid hydrogen sulfate (113 mg) as an amorphous solid. 30 1H-NMR(D2O) 1.61 (6H, s), 2.01 (2H, tt, J=7.6, 7.6Hz), 2.58 (2H, t, J=7.6Hz), 3.07 (2H, t, J=7.6Hz), 3.23 (1H, d, J=18Hz), 3.45 (1H, d, J=13Hz), 3.72 (3H, s), 5.06 (1H, d, J=15.7Hz), 5.25 (1H, d, J=4.8HZ), 5.28 (1H, d, J=15.7Hz), 5.87 (1H, d, J=4.8Hz), 8.03 (1H, s) 35 Preparation 71 tert-Butyl (5-{ [1-methyl-5-(tritylamino) -1H- pyrazol-4-yl] amino}-5-oxopentyl)carbamate The title compound was obtained from 5-[(tert- 102 WO 2004/039814 PCT/JP2003/013684 butoxycarbonyl) amino] pentanoic acid in the same manner as in Preparation 70. 1H-NMR(CDC13) 1.43 (9H, s) , 1.2-1.6 (4H, m) , 1.90 (2H, t, J=7.0Hz), 2.90 (3H, s), 3.09 (2H dt, J=7.0, 7.0Hz), 5 4.52 (lH, s) , 4.61 (1H, t, J=7.OHz), 6.28 (1H, s), 7.0- 7.35 (16H, m), 7.59 (1H, s) I Example 44 3 - ({3-Amino-4- [ {5-aminopentanoyl) amino] -2-methyl- l-pryazolio}methyl)-7-amino -1,2,4-thiadiazol- 10 3-y 1} - 2- (1 -carboxy-l-methylethoxyimino) acetamido] -3 - cephem-4-carboxylic acid hydrogen sulfate The title compound was obtained from tert-butyl (5- {[1-methyl-5- (tritylamino) -lH-pyrazol-4-yl ] amino) -5- oxopentyl) carbamate in the game manner as in Example 43 . 15 1H-NMR(D2O) 1.61 (6H, s) , 1.65-1.8 (4H, m) , 2.50 (2H, m), 3.03 (2H, m), 3.23 (1H, d, J=l6Hz), 3.45 (1H, d, J=l8Hz), 3.72 (3H, s), 5.06 (1H, d, J=15.7Hz), 5.25 (1H, d, J=4.8Hz) , 5.28 (lH, d, J=l5.7Hz), 5.87 (1H, d, J=4.8Hz), 8.02 (lH, s) 20 Preparation 72 To a solution of 1-methyl—N5-trityl-lH-pyrazole- 4,5-diamine (4 g) in dichloromethane (100 ml) was added tert-butyl 4- {[2, 5-dioxo-l-pyrrolidinyl) oxy] carbonyl ]- 1-piperidinecarboxylate (4.05 g), and the mixture was 25 refluxed for 72 hours. The reaction mixture 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 vaciao to give tert-butyl 4- { {1-methy 1-5- 30 (tritylamino)-lH-pyrazol-4-yl]amino]carbonyl)-1- piperidinecarboxylate (1.806 g). 1H-NMR(CDC13) 1.3-1.9 (14H, m) , l.5-1.8 (2H, m) , 2.95 (3H, s), 4.10 (2H, m) , 4.36 (1H,s), 6.53 (1H, s), 7.0-6 7.35 (16H, m) , 7.68 (lH, s) 35 Example_45 3- {(3-amino-2-methyl -4- {(4- piperidinylcarbonyl) amino] -l-pyrazolio}methyl) -7- [ (Z) - 2- (5-amino-l ,2 ,4-thiadiazol-3-yl) -2- (1-carboxy-l- 103 WO 2004/039814 PCT/JP2003/013684 methylethoxyimino) acetamido] -3-cephem-4-carboxylate The title compound was obtained from tet-butyl 4— ({[1-methy1-5-tritylamino)-lH-pyrazol-4- yl]amino}carbonyl) -l-piperidinecarboxylate in the same 5 manner as in Example 36. 1H-NMR(D2O) 1.57 (6H, s) , 1.8-2.3 (4H, m), 2.7-3.6 (7H, m) , 3.72 (3H, s) , 5.06 (1H, d, J=15.7Hz). 5,25 (1H, d, J=4.8Hz), 5.28 (1H, d, J=15.7Hz), 5.37 (1H, d, J=4.8Hz), 8.01 (1H, s) 10 Preparation 73 To a suspension of 3- [N- (tert-butoxycarbonyl )-K- methylamino]propanoic acid (3.33 g) in dicliloromethane (33 ml) and tetrahydrofuran (33 ml) were added HOBT (3.33 g) and WSC HCl (6.29 g) , and the mixture was 15 stirred for 1 hour. To the solution were added 1- methyl-lH-pyrazole-4 ,5-diamine sulfate (3.4 5 g) and N,N- diisopropylethylamine (11.4 ml). The reaction mixture was stirred at room temperature overnight. To the resulting solution was added brine and extracted with 20 tetrahydrofuran/ethyl acetate = 1/1. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give tert-butyl N-{3-[(5-amino- 1—methyl—lH—pyrazol-4—yl) amino] — 3—oxopropyl} —N- methylcarbamate as an oil (2.4 g) . This product was 25 used in the next step without further purification. Preparation 7 4 To a solution of tert-butyl N-{3-{5-amino-l- methyl-lH-pyrazol-4-yl)amine]-3-oxopropyl}-N- [methylcarbamate (4. 88 g) in N, N-dimethylformamide (50 30 ml} were added trityl chloride (6.86 g), triethylamine (6.86 ml) and 4-dimethylaminopyridine (80 mg) succesively. The mixture was stirred at room temperature overnight. To the resulting mixture was added ethyl acetate and washed with water (three times) 35 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 to give tert-butyl N-methyl-N-(3-{[1-methyl— 104 WO 2004/039814 PCT/JP2003/013684 5- (tritylamino) -lH-pyrazol-4-yl] amino} -3- oxopropyl)carbamate (4.20 g) as an amorphous solid. IR(KBr) 1659, 1587, 1491, 1446, 1173, 1151, 762, 739, 708 cm-1 5 1H-NMR(DMSO-d6) 1.40 (9H, s), 2.12 (2H, t, J=7.4Hz), 2.74 (3H,s) , 2.74 (3H, s), 3.24 (2H, t, J=7.4Hz), 5.58 (1H, s), 7.13-7.40 (16H, m), 8.30 (1H, s) Example 46 3- [ [3-Amino-2-methyl-4-{ [3- 10 (methylamino) propanoyl] amino ]-1-pyrazolio)methyl ] -7 [ (Z)-2- (5-amino-l,2,4-thiadiazol-3-yl) -2- (l-carboxy-1- methylethoxyimino) acetamido] -3 -cephem-4-carboxylate The title compound was obtained from tert-butyl N- methyl-N- (3-{ [l-methyl-5- (tritylamino) -lH-pyrazol-4- 15 yl] amino }— 3—oxopropyl) carbamate in the same msnner as in Example 32 as an amorphous solid, IR(KBr) 1770, 1664, 1599, 1531, 1400, 1360 cm-1 1H-NMR(D2O) 1.53 (6H, s) , 2.77 )(3H,s) , 2.92 (2H, t, J=6.5Hz), 3.13 and 3.45 (2H, ABq, J=17.7Hz), 3.74 (3H, 20 s) , 5.00 and 5. 21 (2H, ABq, J=15.4Hz), 5.25 (1H, d, J=4.8Hz), 5.35 (1H, d, J=4.8Hz), 8.02 (1H, s) ESI-MS 666.3 (M+H+) Preparation 7 5 tert-Butyl 3-{ [ {5-amino-l-methyl-lH-pyrazol-4- 25 yl) amino ) carbonyl) -1-azetidinecarboxylate The title compound was obtained from 1-(tert- butoxycarbonyl) -3-azetadinecarboxylic acid in the same manner as in Preparation 73 as an oil. This product was used in the next step without further purification. 30 Preparation 76 tert-Butyl 3-(([l-methyl-5-(tritylamino)-1H- pyrazol-4—yl) amino)carbonyl)-1—azetidinecarboxylate The title compound was obtained from tert-butyl 3- ( [(5-amino-l-methyl-lH-pyrazol-4-yl)amino] carbonyl}-1— 35 acetidinecarboxylate in the same manner as in Preparation 74 as an amorphous solid. IR(KBr) 3367, 3321, 1701, 1662, 1489, 1414, 1144, 766, 704 cm-1 105 WO 2004/039814 PCT/JP2003/013684 1H-NMR(DMSO-d6) 1.39 (9H, s), 2.75 (3H, s), 2.97-3.05 (1H, m) , 3.63-3.70 (2H, m) , 3.82-3.90 (2H, m) , 5.57 (1H, s), 7.10-7.33 (16H, m), 8.41 (1H, s) ESI-MS 560.3 (M+Na+) 5 Example 47 3- ( {3-Amino-4- [ {3-azetidinylcarbonyl) amino] -2- methyl-l-pyrazolio)methyl)-7 [(Z) -2- (5-amino-l, 2 ,4- thiadiazol-3-yl)-2-(1-carboxy-l- methylethoxyimino) acetamido} —3-cephem-4-carboxylate 10 The title compound was obtained from tert-butyl 3- ( {[l-methyl-5-(tritylamino)-lH-pyrazol-4- yl]amino)carbonyl)-l-azetidinecarboxylate in the same manner as in Example 32 as an amorphous solid. IR(KBr) 1768, 1663, 1624, 1605 , 1406, 1362 cm-1 15 1H-NMR(D2O) 1.53 (3H, s), 1.53 (3H, s) , 3.19 and 3.50 (2H, ABq, J=l7.7Hz), 3.82-3.98 (1H, m) , 4.31-4.35 (4H, m) , 4.49 and 5.20 (2H, ABq, J=15.3HZ), 5.25 (1H, d, J=4.8Hz), 5.85 (1H, d, J=4.8Hz), 8.04 (1H, s) ESI-MS 664.2 (M+H+) 20 Preparation 77 tert-Butyl N-(2-[ (5-amino-l-methyl-lH-pyrazol-4- yl) amino] -2-oxoethyl }-N-methylcarbamate The title compound was obtained from [H-(tert- butoxycarbonyl) -N- (methyl)amino] acetic acid in the same 25 manner as in Preparation 73 as an oil. This product was used in the next step without further purification. Preparation 7 6 tert-Butyl N-metyl-N-(2-{[-methyl-5- [tritylamino)-1H-pyrazol-4-yl]amino]-2- 30 oxoethyl)carbamate The title compound was obtained from tert-butyl N- {2-[(5-amino-l-methyl-lH-pyrazol-4-yl)amino]-2- oxoethyl}—N-methylcarbamate in the sams manner as in Preparation 74 as a white solid. The NMR spectrum of 35 this compound indicates the existence of its rotamer, 1H-NMR (DMSO-d6 ) 1.32 and 1.39 (9H,s) , 2.7 2 and 2.77 (3H, s) , 3.52 and 3.61 (2H, brs) , 5.61 (1H, s) , 7.13- 7.33 (16H, m), 8.20 and 8.30 (1H, brs) 106 WO 2004/039814 PCT/JP2003/013684 ESI-MS 548.3 (M+Na+) Example 48 3-[ {3-Amino-2-methyl-4- ([(methylamino)acetyl]amino)-1-pyrazolio)methyl]-7- 5 [ (Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2- (1-carboxy-l- methylethoxyimino} acetamido] -3-cephem-4-carboxylate The title compound was obtained from tert-butyl N- methyl-N- (2-{ [1-methy 1-5- (tritylamino) -lH-pyrazol-4- yl)amino}-2-oxoethyl) carbamate in the same manner as in 10 Example 32 as an amorphous solid. IR(KBr) 1770, 1657, l601, 1400, 1362 cm-1 lH-NMR(D2O) 1.53 (6H, s), 2.82 (3H, s), 3.18 and 3.45 (2H, Abq, J=17.7Hz), 3.74 (3H, s), 4.08 (2H, s), 5.00 and 5.20 (2H, ABq, J=15.3Hz), 5.25 (1H, d, J=4.8Hz), 15 5.84 (lH, d, J=4.8Hz), 8.05 (1H, s) ESI-MS 652.2 (M+H+) Proparation 79 N- (5-Amino-l-methyl-lH-pyrazol-4-yl) -2- (1 , 3-dioxo- 1,3-dihydro-2H-isoindol-2-yl) acetamide 20 The title compound was obtained from (1,3-dioxo- 1,3-dihydro-2H-isoindol-2-yl)acetic acid in the same manner as in Preparation 73 as a solid. 1H-NMR(DMSO-d6) 3.55 (3H, s), 4.36 (2H, S), 4.91 (2H, brs), 7.14 (1H, s), 7.85-8.02 (4H, m), 9.48 (1H, s) 25 ESI-MS 322.2 (M+Na+) Preparation 80 2-(1,3-Dioxo-l,3-dihydro-2H-isoindol-2-yl)-N-[1- methyl-5—(tritylamino)-lH-pyrazol-4—yl]acetamide The title compound was obtained from N-(5-amino-l- 30 methyl-lH-pyrazol-4-yl)-2-{1,3-dioxo-l,3-dihydro-2H- isoindol—2-yl)acetamide in the same manner as in Preparation 74 as aa solid. 1H-NMR(DMSO-d6) 2.70 (3H, s) , 4.12 (2H, 3) , 5.41 (1H, S), 7.12-7.33 (16H, m) , 7.85-7.95 (4H, m), 8.93 (1H, s) 35 ESI-MS 564.3 (M+Na+) Preparation 81 Hydrazine monohydrate (1.46 ml) was added to a solution of 2-{1,3-dioxo-l,3-dihydro-2H-isoindol-3-yl)- 107 WO 2004/039814 PCT/JP2003/013684 N-[l-methyl-5-(tritylamin)-lH-pyrazol-4-yl]acetamide (5.42 g) in ethanol (108 ml) and tetrahydrofuran (54 ml) at room temperature, and the mixture was stirred at 70°C for 2 hours. The reaction mixture was cooled to 0°C, 5 and the insoluble materials were removed by filtration. The filtrate was concentrated in vacuo. The residue was triturated with diisopropyl ether, collected by filtration and dried in vacuo to give 2-amino-N-[1- methyl-5-(tritylamino)-lH-pyrazo1-4-yl]acetatmide (3.3 7 10 g) as a solid. This product was used in the next step without further purification. Preparation 82 To a solution of 2—amino-N- [l-methyl-5- (tritylamino)-lH-pyrazol-4-yl]acetamide (2.47 g) in 15 tetrahydrofuran (50 ml) were added di-tert-butyl ({[(trifluoromethyl)sulfonyl]imino}metiiylene) - biscarbamate (2.35 g) and triethylamine (2.5 ml), and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was poured into a mixture 20 of ethyl acetate and water. The aqueous layer was separated, and the organic layer was washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated in vacuo. The concentrate was purified by silica gel column chromatography to give 25 di-tert-butyl { (E) -[(2-{[l-methyl-5-(tritylamino)-1H- pyrazol-4-yl] amino}-2-oxoethyl)amino]methylidene}- biscarbamate (3.25 g) as an amorphous solid. 1H-NMR(DMSO-d6) 1.38 (9H, s), 1.49 (9H, s), 2.75 (3H, s), 3.79 (2H, d, J=4.7Hz), 5.47 (1H, s), 7.12-7.33 (16H, 30 m), 8.55 (1H, t, J=4.7Hz), 8.61 (1H, s), 11.43 (1H, s) ESI-MS 676.3 (M+Na+) Example 49 3- ({3-Amino—4— [ (guanidinoacetyl) amino] —2—methyl—1- pyrazolio}methyl)-7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3- 35 yl) -2- (1-carboxy-l-methylethoxyimino) acetamido] -3- cephem- 4—carboxy1ate The title compound was obtained from di-tert-butyl { {E)-[ (2-{l-methyl-5-(tritylamino)-lH-pyrazol-4- 108 WO 2004/039814 PCT/JP2003/013684 yl] amino} -2-oxoethyl) ami no] methylidene )biscarbamate in the same manner as in Example 32 as an amorphous solid. IR(KBr) 1770, 1668, 1655, 1620, 1601, 1402, 1363 cm-1 1H-KMR(D2O) 1.53 (6H, s) , 3.20 and 3.48 (2H, ABq, 5 J=l7.6Hz, 3.75 (3H, s) , 4.21 (2H, s) , 5.00 and 5 . 20 (2H, ABq, J=15.3Hz) , 5.26 (1H, d, J=4 . 8Hz) , 5.85 (1H, d, J=4.8Hz) , 8.02 (1H, s) ESI-MS 678.2 (M-H+) (negative) Example 50 10 To a solution of 3-((3-amino-4-[(3- aminopropanoyl) amino]—2—methyl-1—pyrazolio }methyl- 7- [(Z) -2- [5-amino-l, 2 , 4-thiadiazol-3-yl) -2- (1-carboxy-l- methylethoxyimino)acetamido]-3-cephem-4-carboxylate (652 mg) in water (30 ml) and acetonitrile (3 ml) were added 15 ethyl formimidate hydrochloride (6 58 mg) and potassium carbonate (1.106 g) under ice cooling. After stirring at 5°C, for 3 hours , 1N HCl was added to neutralize the reaction mixture. The resulting solution was purified by preparative HPLC eluting with a mixture of phosphate 20 buffer (pH 5.5) and acetonitrile, and the eluate was subjected to column chromatography on Diaion© HP20 (Mitsubishi Chemical Corporation) and lyophilized to give 3- ( {3-amino-4- [ (3-guanidinopropanoyl) amino] -2- methyl-1-pyrazolio}methyl) -7[ (Z) -2- (5-amino-l , 2 ,4- 25 thiadiazol-3-yl)-2-(1-carboxy-l- methylethoxyimino) acetamidol]-3-cephem-4-carboxylate (23 mg) as an amorphous. The NMR spectrum of this compound indicates the existence of its rotamer. Only major isomsr was described. 30 IR(KBr) 1770, 1714, 1668, 1653, 1456, 1400 , 1360 cm-1 1H-NMR(D2O) 1.53 (6H, s), 2.85 (2H, t, J=6.4Hz), 3.19 and 3.46 (2H, ABq, J=l7.7Hz), 3.65 (2H , t, J=6.4Hz), 5.00 and 5.21 (2H, ABq, J=l5.2Hz), 5.26 (1H, d, J=4.8Hz), 5.85 (1H, d, J=4.8Hz), 7.80 (1H, s), 8.01 (1H, s) 35 ESl-MS 677.2 (M-H+) (negative) Preparation 83 To a stirred solution of l-methyl-lH-pyrazole-4,5- diamine sulfate (2.1 g) and 3-ethoxy-3-oxopropanoic acid. 109 WO 2004/039814 PCT/JP2003/013684 (1.32 g) in dichloromethane (10 ml) and tetrahydrofuran (10 ml) was added WSCD HCl (3.83 g) and N,N- diisopropylethylamine (6.96 ml), and the mixture was stirred overnight. The solvent was removed under 5 reduced pressure, and the crude residue which includes ethyl 3-[(5-amino-l-methyl-lH-pyrazol-4-yl)amino]-3- oxopropanoate was used for the next reaction without further purification. Preparation 84 10 The crude residue containing ethyl 3-[ (5-amino-l- methyl—1H—pyrazol-4-yl) amino] — 3-oxopropanoate was dissolved in N,N-dimethylformamide (20 ml), and trityl chloride (5.52 g) and triethylamine (4.14 ml) were added with stirring. The mixture was stirred overnight and 15 quenched with water (10 ml). The whole mixture was extracted with ethyl acetate, and the extract was washed with water and brine, dried over magnesium sulfate and concentrated, under reduced pressure to give a residual oil, which was chromatographed on silica gel eluting 20 with dichloromethane-ethyl acetate (2:3) to give ethyl 3-{[l-methyl-5-(tritylamino)-lH-pyrazol-4-yl]amino}-3- oxopropanoate (1,23 g) . ESI-MS 491.2 [M+Na]+ (positive), 467.3 (M-H]- (negative) 1H-NMR(DMSO-d6) 1.13 (3H, t, J=7.1Hz), 2.75 (3H, s), 25 3.04 (2H, s), 4.07 (2H, q, J=7.1Hz), 5.55 (1H, s), 7.1- 7.4 (16H, m), 8.54 (1H, s) Preparation 8 5 To a stirred solution of ethyl 3-[[l-methyl-5- (tritylamino) —lH-pyrazol-4-yll amino]-3-oxopropanoate 30 (1.3 g) in tetrahydrofuran (30 ml) was added 1N aqueous sodium hydroxide solution (3.1 ml), and the mixture was| stirred at room temperature for 3 hours. Tetrahydrofuran was removed in vacuo and the residue was made acidic with diluted citric acid. The resulting 35 precipitate was collected by filtration and dried under reduced pressure to give 3-{ (l-methyl-5-(tritylamino)- lH-pyrazol-4-yl] amino}-3-oxopropanoic acid (1.22 g) . ESI-MS 463.2 [M+Na]+ (positive) 110 WO 2004/039814 PCT/JP2003/013684 1H-NMR(DMSO-d6) 2.74 (3H s) , 2.95 ,(2H,s), 5.56 (1H, s) , 7.0-7.4 (16H, m), 8.54 (1H, s) , 12.0-13.0 (1H, brs) Preparation _86 To a suspension of 3-{ [l-methyl-5-(tritylamino)- 5 lH-pyrazol-4-yl) amino}-3-oxopropanoic acid (600 mg) and tert-butyl (2-aminoethyl)carbamate (240 mg) in tetrahydrofuran (12 ml) and dichloromethane (6 ml) was added WSCD HC1 (522 mg), and the whole mixture was stirred at room temperature overnight. To the reaction 10 mixturc was added water (3 ml) , and the whole mixture was extracted with ethyl acetate. The extract was washed with water and brine and dried over magnesium sulfate. The evaporation of the solvent gave a crude residue/ which was triturated with diisopropyl ether- 15 ethyl acetate (2:1) to give tert-butyl {2-[(3-{[1- methyl -5- (tritylamino ) -1H-pyrazol-4 -y 1 ] amino) -2- oxopropanoyl] amino] ethyl ) carbamate (53 7 mg) . ESI-MS 604.3 [M+Na]+ (positive) 1H-KMR(DMSO-d6) 1.38 (9H, s) , 2.74 (3H, s), 2 . 85 (2H, 20 s) , 2.9-3.2 (4H, m) , 5.61 (lH, s) , 6.7-6.3 (1H, m) , 7.0- 7.4 (16H, m), 8.0-8.1 (1H, m), 8.63 (lH, s) Example 51 3-{[3-Amino-4- [{3-[( 2-aminoethyl) amino}-3- oxopropanoyl] amino] - 2-methyl - 1-pyrazolio] methyl) –7b- 25 [ (Z) -2-(5-amino-l,2,4-thiadiazol-3-yl}-2-{l-carboxy-l- methylethoxyimino) acetamido] -3-cephem-4-carboxylate The title compound was obtained from tert-butyl {2-[{3-{[1-methyl-5-(tritylamino)-lH-pyrazo1-4- yl ]amino}-3-oxopropanoyl) amino] ethyl} carbamate in the 30 same manner as in Example 34. ESI-MS 731.2 [M+Na]+ (positive) 1H-NMR(D2O) 1.53 (6H, S) , 3.1-3.3 (2H, m), 3.19 and 3.44 (2H, ABq, J=l7.7Hz), 3. 54 (2H, s), 3.5-3.7 (2H, m) , 3.74 (3H, s) , 5.00 and 5.22 (2H, ABq, J=l5.5Hz), 5.25 35 (lH, d, J=4.7Hz), 5.86 (lH, d, J=4.8Hz), 8.05 (lH, s) Preparation 87 To a stirred solution of 3-amino-2- hydroxypropanoic acid (2.1 g) in tetrahydrofuran (30 ml) 111 WO 2004/039814 PCT/JP2003/013684 and water (30 ml) was added 1H aqueous sodium hydroxide solution to make the solution basic (pH=9) . To the mixture was added di-tert-butyl dicarbonate (4.36 g) , and the mixture was stirred at room temperature for 4 5 hours keeping pH of the mixture between 8.5 and 9.0. The whole mixture was washed with diethyl ether. The aquous layer was made acidic (pH=2) with 10% aqueous potassium hydrogen sulfate, saturated with sodium chloride and extracted with ethyl acetate. The extract 10 was dried over magnesium sulfate. The solvent was evaporated under reduced pressure to give 3-[(tert- butoxycarbonyl) amino]-2-hydroxypropanoic acid (3.96 g) . ESI-MS 228.2 [M+Na]+ (positive) 1H-NMR(DMSO-d6) 1.37 (9H, s), 3.0-3.8 (3H, m) , 3.9-4.1 15 (1H, m), 6.5-6.8 (lH, m) Preparation 88 To a solution of 3—[ (tert—butoxycarbonyl)amino] — 2 — hydroxypropanoic acid (1.61 gl in dichloromethane (8 ml) and tetrahydrofuran (8 ml) were added HOBT (1.59 g) and 20 WSCD HC1 (3.01 g) , and the mixture was stirred at room temperature for 1 hour. The solution was cooled to 0°C, and 1-methyl-lH-pyrazole—4 ,5-diamine sulfate and N,N— diisopropylethylamine (4.1 ml) were added. The mixture was stirred at room temperature for 8 hours. The 25 solvent was removed under reduced pressure to give crude tert-butyl {3—[(5—amino-1-methyl-1H-pyirazol-4-yl)amino]- 2-hydroxy-3-oxopropyl)carbamate, which was used in the next reaction without further purification. Preparation 89 30 tert-Butyl (2-hydroxy-3- { [l-methyl-5- (tritylamino)-lH-pyrazol-4-yl]amino}-3- oxopropyl)carbamate The title compound was obtained from tert-butyl [3- [ (5-amino-l-methyl-lH-pyrazol-4-yl) amino] -2-hydroxy- 35 3-oxopropyl}carbamate in the same manner as in Preparation 84. ESI-MS 564.3 [M+Na]+ (positive) 1H-NMR (DMSO-d6) 1.39 (9H, s) , 2.7-2.9 (1H, m), 2.83 (3H, 112 I WO 2004/039814 PCT/JP2003/013684 s), 3,1-3.4 (1H, m), 3.7-3.9 (1H, m), 5.79 (1H, d J=5.3Hz), 5.96 (lH, s), 6.5-6.7 (1H, m), 7.1-7.4 (16H, m) , 8.36 (1H, s) Example 52 5 3— { {3-Amino-4- [ (3-amino-l-hydroxypropanoyl} amino- 2-methyl-l-pyrazolio}methyl) - 7 - { (Z) -2- (5-amino-l ,2,4- thiadiazol-3-yl) -2- (1-carboxy-1- methylethoxyimino) acetamido] -3-cephem-4-carboxylate The title compound was obtained from tert-butyl 10 (2-hydroxy-3-{[l-methyl-5-(tritylaminol)-lH-pyrazol-4- yl] amino}-3-oxopropyl) carbamate in the same manner as in Example 32. 1H-NMR(D20) 1.49 (6H, s), 3.1-3.6 (4H, m) , 3.76 (3H, s), 4.6-4.7 (1H, m), 5.02 and 5.21 (2H, ABq, J=l5.4Hz), 5.26 15 (1H, d, J=4.8Hz), 5.86 (1H, d, J=4.8Hz) , 8.05 (1H, s) Preparation 90 To a suspension of 2- (4 , 5-dLamino-1H-pyrazol-l- yl) ethanol sulfate (5 g) in dichlorotmethane (50 ml] was added triethylamine (6.38 ml) at 0oC, and the mixture 20 was stirred at 0oC for 10 minutes. A mixture of acetic anhydride (2.16 ml) and formic acid (1.74 ml) was stirred at 40oC for 30 minutes, cooled to 0°C and added dropwise to the above solution at 0oC. The whole mixture was stirred at 0°C for 2 hours. To the mixture 25 was added brine, and the whole mixture was extracted with tetrahydrofuran. The extract was dried over magnesium sulfate and evaporated under reduced pressure to give crude [5-amino-l- (2-hydroxyethyl) -lH-pyrazol-4- yl]formamide, which was used in the next reaction 30 without further purification. Preparation_91 [1- (2-Hydroxyethyl) -5- (tritylamino) -lH-pyrazol-4- yl] formamide The title compound was obtained from [5-amino-l- 35 (2-hydroxyethyl)-lH-pyrazol-4-yl]formamide in the same manner as in preparation 84. The NMR spectrum of this compound indicates the existence of its rotamer. 1H-NMR(DMSO-d6) 3.10 (2H, t, J=6.2Hz), 3.3-3.5 and 3.4- 113 WO 2004/039814 PCT/JP2003/013684 3.6 (2H, m), 4.89 and 5.06 (1H, t, J=5.lHz), 5.77 and 6.07 (1H, s) , 7.1-7.4 (16H, m) , 7.58 and 8.07 (1H, s) , 7.58 (1H, s) Preparation 92 5 To a stirred solution of [1-(2-hydroxyethyl)-5— (tritylamino)-lH-pyrazol-4-yl]formamide (2 g) in N,N- dimethylformamide (30 ml) was added sodium hydride (213 mg, 60% oil suspension) under a nitrogen stream at 0°C, and the whole mixture was stirred at 0oC for 20 minutes. 10 A solution of tert-butyl (3-bromopropyl) carbamate (1.27 g) in N, N-dimethylformamide (10 ml) and sodium iodide (799 mg) were added to the above solution, and the mixture was stirred overnight. 10% Aqueous potassium hydrogen sulfate solution (5 ml) was added, and the 15 whole mixture was extracted with ethyl acetate. The extract was washed with water and brine and dried over magnesium sulfate. Evaporation of the solvent under reduced pressure gave an oil, which was chromatographed on silica gel eluting with dichloromethane-ethyl acetate 20 (2:1) to give tert-butyl (3-(N-formyl-N-[1-(2- hydroxyethyl)—5—(tritylamino)-1H—pyrazol—4— yl ] amino }propyl) carbamate (1 g) . The NMR spectrum of this compound indicates the existence of its rotamer. ESI-MS 592.3 [M+Na]+ (positive) 25 1H-NMR(DMSO-d6) 1.37 and 1.38 (9H, g) , 2.7-3.5 )(10H, m), 4.80 and 4.88 (lH, t, J=5.0Hz), 5.52 & 6.06 (1H, s), 6.5-6.9 (1H, m), 7.0-7.4 (16H, m), 7.52 (1H, s) Example 53 3— {{ 3—Amino-4- [N- (3-aminopropyl) -N-formylaminol -2- 30 {2-hydroxyethyl) -1-pyrazolio)methyl) -7- [ (Z) -2- (5-amino- l,2,4-thiadiazol-3-yl)-2-(1-carboxy-l- methylethoxyimirLO)acetamido]—3-csphem—4-carboxylate The title compound was obtained from tert-butyl (3-[N-formyl-H-[l-(2-hydroxyethyl)-5-(tritylamino)-1H- 35 pyrazol—4—yl]amino}propyl)carbamate in the same manner as in Example 32. ESI-MS 694.2 [M-H]- (negetive) 1H-NMR(D2O) 1.53 (6H, s) , 1.7-2.1 (2H, m) , 2.9-3.1 (2H, 114 WO 2004/039814 PCT/JP2003/013684 m) , 3.1-3.8 (4H, m) , 3.3-4.0 (2H, m) , 4.3-4 .6 (2H, m) , 4.3-5.2 (2H, m) , 5.29 (lH, d, J=4.8Hz), 5.85 (1H, d, J=4.7HZ), 8.0-8.3 (2H, m ) Example__54 . 5 To a stirred suspension of 3-({3-amino-4-[N-(3- aminopropyl) -N-formylamino]-2- (2-hydroxyethyl) -1- pyrazolio}methyl}-7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3- yl) -2- {1-carboxy-l-methylethoxyimino) acetamido] -3- cephem-4-carboxylate (100 mg) in methanol (l.4 ml) was 10 added concentrated hydrochloric acid (0.125 ml) at room temperature , and the mixture was stirred for 6.5 hours. To the above solution was added sodium hydrogen carbonate (109 mg), and the mixture was purified by preparative HPLC (ODS column; acetonitrile:phosphate 15 buffer (pH 7) = 5:95). The eluate containing a desired product was evaporated to remove acetonitrile, made acidic with diluted hydrochloric acid and chromatographed on Diaion® HP2 0 (Mitsubishi Chemical Corporation) eluting with 20% aqueous 2-propanol. The 20 eluate was concentrated under reduced pressure and lyophilized to give 3-{{3-amino-4-[(3- aminopropyl)amino)-2-(2-hydroxyethyl)-1- pyrazolio}methyl)-7-[ (Z) -2- (5-amino-l , 2 , 4-thiadiazol-3- yl)-2—(1-carboxy-l—methylethoxyimino)acetamido]-3- 25 cephem-4-carboxylate (18 mg) . ESI-MS 666.2 [M-H]- (negative) 1H-NMR(DMSO-d6) 1.53 (6H, s) , 1.96 (2H, tt, J=7.5Hz), 3.0-3,Z (4H, m), 3.13 and 3.43 (2H, ABq, J=17.6Hz), 3.87 (2H, t, J=4.8Hz), 4.2-4.4 (2H, m) , 4.87 and 5.03 (2H, 30 ABq, J=15.2Hz), 5.24 (1H, d, J=4.8Hz), 5.8 3 (lH, d, J=4.8 Hz) , 7.64 (1H, s) Preparation 93 To a stirred solution of N-[1-methyl-5- (tritylamino)-lH-pyrazol-4-yl]-2-(tritylamino)acetamide 35 (2 g) in N,N-dimethylformamide (20 ml) was added sodium hydride (245 mg, 60% oil suspension) at 0oC, and the mixture was stirred for 30 minutes with warming to room temperature. The mixture was cooled to 0oC, and methyl 115 WO 2004/039814 PCT/JP2003/013684 iodide (1.3 g) was added. The whole mixture was stirred at room temperature overnight. Water (5 ml) was added, and the whole mixture was extracted with ethyl acetate. The organic layer was washed with, water and birine and 5 dried over magnesium sulfate. Evaporation of the solvent under reduced pressure gave N-methyl—H-[1—methyl-5— (tritylamino)-lH-pyrazol-4-yl]-2-(tritylamino)acetamide (2.05 g). ESI-MS 690.3 [M+Na]+ (positive) 10 1H-NMR(DMSO-d6) 1.99 (3H, s), 2.3-2.S (3H, m) , 2.52 (3H, s), 5.44 (1H, s) , 6.85 (1H, s), 6.9-7.5 (30H, m) Preparation 94 Lithium aluminum hydride (4 55 mg) was added slowly to tetrahydrofuran (40 ml) at OoC and the mixture was 15 stirred for 20 minutes, N-Methyl-N-[l-methyl-5- (tritylamino)-lH-pyrazol-4-yl]-2-(tritylamino)acetamide (2 g) was added to the mixture at 0oC, and the whole mixture was stirred for 2 hours with warming to room temperature and refluxed for 2 hours. Sodium fluoride 20 (2.51 g) and water (8 62 mg) were added to the mixture, and the whole mixture was stirred at room temperature for 30 minutes. The precipitate was filtered off, and the filtrate was concentrated under reduced pressure to give crude residue, which was chromatographed (silica 25 gel; ethyl acetate:dichloromethane=l:10) to give N4,1- dimethyl-N5trityl-N4- [2- (tritylamino) ethyl] -1H- pyrazole-4,5-diamine (740 mg) . ESI-MS 676.2 [M+Na]+ (positive) 1H-NMR(DMSO-d6) 1.7-2.0 (2H, m) , 1.98 (3H, s), 2.2-2.4 30 (lH, m) , 2.6-2.8 (2H, m) , 2.81 (3H, s), , 5.24 (1H, s), 7.00 (1H, s), 7.0-7.5 (30H, m) Example 55 3- ( {3-Amino-4- [N- (2-aminoethyl) -N-methylamino] -2- methyl-l-pyrazolio}methyl)-7-[(Z)-2-(5-amino-l,2,4- 35 thiadiazol-3-yl)-2-(1-carboxy-l- methy1ethoxyimino)acetamido]-3-cephem-4-carboxy1ate The title compound was obtained from N4,l- dimethyl-N5-trityl-N4-[2-(tritylamino) ethyl] -lH- 116 WO 2004/039814 PCT/JP2003/013684 pyrazole-4, 5-diamine in the same manner as in Example 32. ESI-MS 636.2 [M-H]- (negative) 1H-NMR(D2O) 1.60 (6H, S) , 2.60 (3H, s), 3.0-3.2 (4H, m), 3.19 and 3.39 (2H, ABq, J=l7.7Hz), 3.67 (3H, s) 4.87 5 and 5.20 (2H, ABq, J=15.8Hz), 5.22 (1H, d, J=4.9Hz), 5.85 (1H, d, J=4.7Hz), 7.90 (1H, s) Preparation 95 To a solution of [1-(2-fluoroethyl)-lH-pyrazol-5- yl] formamide (15.7 g) in methanol (7 8 ml) was added 10 concentrated hydrochloric acid (21 ml) at room temperature. The reaction mixture was stirred for 3.5 hours and evaporated in vacuo . The residue was dissolved in ethyl acetate and washed with aqueous sodium hydrogen carbonate solution. The organic layer 15 was dried over magnesium sulfate and concentrated in vacuo to give 1- (2-fluoroethyl) -lH-pyrazol-5-amine (12 g). 1H-NMR(DMSO-d6) 4 . 15 (2H, dt, J=25 . 2 , 5.1HZ), 4.66 (2H, dt, J=47.2, 5.lHz), 5.1 (2H, brs) , 5.27 (lH, d, J=1.7HZ), 20 7.O6 (1H, d, J=1.7Hz) Preparation _9_6 To a solution of 1-(2-fluoroethyl)-lH-pyrazol-5- amine (12 g) in ethanol (30 ml) were added concentrated hydrochloric acid (70 mg) and isoamyl nitrite (10.9 g), 25 The reaction mixture was stirred at 25-38oC for 2 hours. Diisopropyl ether and hexane were added to the reaction mixture, and the resulting oil was purified by column chromatography on silica gel (ethyl acetate :hexane=l:2 à1:1 2:1 1:0) to give 1- (2-fluoroethyl) -4- 30 nitroso-lH-pyrazol-5-amine (4.8 g). 1H-NMR(DMSO-d6) 4.10-4.90 (4H, m), 7.09 and 8.59 (1H, s), 8.20 and 8.26 (1H, brs) Preparation 97 To a solution of 1-(2-fluoroethyl)-4—nitroso-lH— 35 pyrazol-5-amine (4,8 g) in water (30 ml) and methanol (30 ml) were added sulfuric acid (2.98 g) and 10% palladium on carbon (2.5 g), and the mixture was hydrogenatad under balloon pressure for 7.5 hours. The 117 WO 2004/039814 PCT/JP2003/013684 reaction mixture was filtered through a bed of Colite, and the filtrate was concentrated in vacuo. 2-Propanol was added to the residue, and the precipitate was collected by filtration to give 1-(2-fluoroethyl)-1H- 5 pyrazole-4 ,5-diamine sulfate (7 g). 1H-NMR(D2O) 4.25-4.95 (4H, m), 7.66 (1H, s) Preparation 9 8 To a suspension of 1-[2-fluoroethyl)-lH-pyrazole- 4,5-diamine sulfate (3 g) in tetrahydrofuran (30 ml) 10 were added tert-butyl {3-[(2,5-dioxo-l- pyrrolidinyl)oxy]-3—oxopropyl)carbamate (3.9 g) and ,N,N— diisoporpylethylamine (3.5 g) under ice-cooling. The reaction mixture was stirred at room temperature for 2 hours. An aqueous sodium hydrogen carbonate solution 15 and sodium chloride were added, and the mixture was extracted with ethyl acetate-tetrahydrofuran (three times). The organic layer was dried over magnesium sulfate and concentrated in vacuo. The residue was purified by column chromatography on silica gel (ethyl 20 acetate ethyl acetate:ethanol=8:1) to give tert-butyl (3-{ [5-amino-l- (2-fluoroethyl) -1H-pyrazol-4-yl] amino)-3- oxopropyl] carbamate (2.3 g) . 1H-NMR(DMSO-d6) 1.33 (9H, s), 2.36 (2H, t, J=7.1Hz), 3.10-3.27 (2H, m), 4.16 (2H, dt, J=25.5, 5.0Hz),4.67 (2H, 25 dt, =47.2, 5.OHz), 5.27 (2H, brs), 6.75-6.90 (1H, m) , 7.23 (1H, s), 9.03 (1H, brs) Preparation 99 To a solution of tert-butyl (3-{[5-amino-l-(2- fluoroethyl)-lH-pyrazol-4-yl]amino}-3— 30 oxopropyl) carbamate (2.3 g) in N,N-dimethylfomamide (12 ml) were added triethylamine (1.48 g), 4- dimethylaminopyridine (35.6 mg) and trityl chloride (2.2 g) at room temperature. The reaction mixture was stirred for 2 hours, and water was added. The mixture 35 was extracted with ethyl acetate, and the organic layer was washed with, water and aqueous sodium chloride solution. The organic layer was dried over magnesium sulfate and concentrated in vacuo. Acetonitrile was 118 WO 2004/039814 PCT/JP2003/013684 added, and the precipitate was collected by filtration to give tert-butyl {3-([l-(2-fluoroethyl)-5- ( tritylamino)-1H-pyrazol-4-yl] amino }-3- oxopropyl)carbamate (2 g) . 5 1H-NMR(DMSO-d6) 1.39 (9H, s), 2.05 (2H, t, J=7.2Hz), 3.00-3.03 (2H, m), 3.23 (2H, dt, J=25.3, 5.1Hz), 4.41 (2H, dt, J=47.1, 5.1Hz) Example 56 3- {( 3-Amino-4- [(3-aminopropionyl) amino] -2- {2- 10 fluroethyl)-l-pyrazolio)methyl)-7-[ (Z)-2-(5-amino- 1,2,4-thiadiazol-3-yl}-2-(1-carboxy-l- methylethoxyimino) acetamido ] -3-cephem-4 - carboxy1ate The title compound was obtained from tert-butyl (3- {(1-(2-fluroethyl)-5- (tritylamino) -lH-pyrazol-4- 15 yl] amino }-3-oxopropyl) carbamate in the same manner as in Example 38. 1H-NMR(D2O) 2.89 (2H, t, J=6.5Hz), 3.22 (1H, d, J=9.2Hz), 3.34 (1H, t, J=6.5Hz), 3.50 (1H, d, J=9.2Hz), 4.55-4.95 (4H, m), 5.08 (2H, brs), 5.26 (1H, d, J=4.9Hz), 20 5.84 (1H, d, J=4.9Hz), 8.09(1H,s) Preparation 100 l-Methyl-7-nitroso-lH-imidazo [1, 2-b]pyrazole The title compound was obtained from 1-methyl-lH- imidazo [1,2-b]pyrazole in the same manner as in 25 Preparation 96. 1H-NMR(DMSO-d6) 3.93 (1H, s), 7.43 (1H, m) , 7.92 (1H, m), 9.03 (1H, s) Preparation 101 1 -Methyl-lH-imidazo[l,2-b]pyrazol-7- amine sulfate 30 The title compound was obtained from l-methyl-7- nitroso-1H-imidazo[l,2-b]pyrazole in the same manner as in Preparation 97. 1H-NMR (DMSO-d6) 3.73 (3H, s) , 7.24 (1H, m), 7.62 (2H, m) 35 Preparation 102 Di-tert-butyl 1(Z)-[(1-methyl-lH-imidazo[1,2- b]pyrazol-7-yl) amino]methylidene}biscarbamate The title compound was obtained from 1—methyl-lH- 119 WO 2004/039814 PCT/JP2003/013684 imidazo [ 1,2-b] pyrazol-7-amine sulfate in the same manner as in Preparation 64. s) , 7.14 (1H, m), 7.42 (1H, m), 7.52 (1H, m) 5 Example 57 7 b -[ (Z)-2- (5-Amino-l,2,4-thiadiazo-l-3-yl)-2- (1- carboxy-l-methylethoxyimino) acetamido]-3-{ [7-guanidino- l-methyl-5- (lH-imidazo [1, 2-b ] pyrazolio) ] methyl ) -3- cephem-4-carboxylate 10 The title compound was obtained from di-tert-butyl {(Z)-[{1-methyl-lH-imidazo[l,2-b]pyrazol-7- yl) aminomethylidene } biscarbamate in the same manner as in Example 43. 1H-NMR(D2O) 1.51 (6H, s), 3.40 (2H, m), 3.85 (3H, s), 15 5.15-5.30 (3H, m) , 5.83 (1H, d, J=4.8Hz), 7.49 (1H, d, J=2.2Hz), 8.02 (1H, d, J=2.2Hz), 8.27 (1H, d, J=1.0Hz) IR(KBr) 3400, 3392, 1770, 1672, 1606, 1531 cm-1 preparation 103 tert-Butyl {3-[ {1-methyl-lH-imidazo [1 , 2-b]pyrzol- 20 7-yl)amino]-3-oxopropyl)carbamate The title compound was obtained from l-methyl-lH- imidazol[1,2-b]pyrazol-7-amine sulfate and 3-[(tert- butoxycarbonyl)amino]propanoic acid in the same manner as in Preparation 70 25 1H-NMR(DMSO-d6) 1.43 (9H, s), 2.61 (2H, m) , 3.49 (2H, m), 3.65 (3H, s), 7.22 (1H, m), 7.26 (1H, m), 7.44 (1H, Example 58 3-|{7-[(3-Aminopropanoyl)amino]-1-methy1-5-(1H- 30 imidazo[1,2-b]pyrazolio) }methyl)-7-[(Z) -2-(5-amino- l,2,4-thiadiazol-3-yl) -2- (1-carboxy-l- methylethoxyimino) acetamido] -3-cephem-4-carboxylate The title compound was obtained from tert-butyl { 3- [ (1-methyl-lH-imidazo [1 ,2-b)]pyrazol-7-yl) amino] -3- 35 oxopropylcarbamate in the same manner as in Example 43. 1H-NMR(D2O) 1.50 (6H, s), 2.97 (2H, d, J=6.5Hz), 3.36 (2H, d, J=6.5Hz), 3.4 (2H, m), 3.81 (3H, s), 5.15-5.30 (3H, m) , 5.82 (1H, d, J=4.8Hz), 7.44 (1H, d, J=2.2Hz), 120 WO 2004/039814 PCT/JP2003/013684 7.98 (1H, d, J=2.2Hz), 3.11 (1H, d, J=1.0Hz) IR(KBr) 3401, 1770, 1665, 1606, 1525 cm-1 Preparation _l04 To a suspension of phenyl [l-methyl-5- 5 {tritylamino) -lH-pyrazol-4-yl] carbamate (4.6 g) in N,N- dimethylformamide (32 ml) were added triethylamine (1.08 g) and tert-butyl l-piperazinecarboxylate (1.99 g). The reaction mixture was stirred for 3 hours and poured into water. The mixture was extracted with ethyl acetate. 10 and the organic layer was concentrated in vacuo. The residue was purified by column chromatagraphy on silica gel (ethyl acetate ethyl acetateethanol=20:1) to give tert-butyl 4- ( { [1-methyl-5- (tritylamino) -1H- pyrazol-4-yl ] amino } carbonyl) -1-piperazinecarboxylate 15 (4.7 g). 1H-NMR(CDCl3) 1.46 (9H, s) , 2.90 (3H, s), 3.05-3.25 (4H, m) , 3.30-3.45 (4H, m) , 4.76 (1H, brs), 5.34 (1H, brs), 7.10-7.30 (l6H, m) Example_59 20 To a solution of 4-methoxybenzyl 7- [ (Z) -2- (5- amino-1, 2 , 4-thiadiazol-3-yl) -2- (l-tert-butoxycarbonyl.-l- methylethoxyimino) acetamido] -3-chloromethyl-3-cephem-4- carboxylate (2 g) in N,N-dimethylformamide (6 ml) was added 1, 3-bis (trimethylsilyl) urea (3 g) , and the 25 reaction mixture was stirred for 30 minutes. Potassium iodide (680 mg) was added to this solution, and the mixture was stirred for 30 minutes, tert-Butyl 4-({1- methyl—5-(tritylamino)-lH-pyrazol-4-yl)amino}carbonyl)- 1-piperazinecarboxylate (2 g) was added to this solution 30 The reaction mixture was stirred at 25oC for 23 hours nd poured into a mixture of ethyl acetate-water-20% queous sodium chloride solution. The organic layer was ashed with a mixture of 10% aqueous sodium thiosulfate olution and 20% aqueous sodium chloride solution. The 35 organic layer was washed successively with 10% aqueous sodium trifluoroacetate solution twice and 20% aqueous sodium chloride solution. The organic layer was concentrated in vacuo to a volume of approximately 10 ml 121 WO 2004/039814 PCT/JP2003/013684 The concentrate was added to diisopropyl ether, and the suspension was stirred for 1 hour, The resulting solid was collected by filtration and dried. The solid was dissolved in dichlromethane (6 ml) . 5 To this solution was added anisole (2 ml) and trifluoroacetic acid (6 ml). The reaction mixture was stirred for 4 hours and poured into diisopropyl ether. The resulting solid was collected by filtration and dried. This solid was purified by preparative HPLC 10 utilizing ODS column. The eluate containing a desired product was concentrated in vacuo. The concentrate was adjusted to about pH 1 with concentrated hydrochloric acid and chromatogaphed on Diaion® HP—20 (Mitsubishi Chemical Corporation) eluting with 20% aqueous 2- 15 propanol. The eluate was concentrated in vacuo, and 2H sulfuric acid was added. The mixture was lyophilizedL to give 3- ( (3-amino-2-methyl-4- [ (1- piperazinylcarbonyl) amino] -1-pyrasolio )methyl) -7- [ (Z) - 2- (5-amino-l ,2 , 4-thiadiazol-3-yl) -2- (1-carboxy-l- 20 methylethoxyimino)acetamido]-3-cephhem-4-carboxylic acid hydrogen sulfate (679 mg) . 1H-NMR(D2O) 1.60 (6H, s) , 3.20 (2H, d, J=17.7Hz), 3.25- 3.45 (4H, m) , 3.45 (1H, d, J=17.7Hz) 3.72 (3H, m) . 3.75- 3.85 (4H, m), 5.00 (1H, d, J=15.7Hz), 5.24 (1H, d, 25 J=15.7HZ), 5.25 (1H, d, J=4.8Hz), 5.86(1H, d, J=4.8Hz) 7.89 (1H, s) This application is based on application No. 2002952355 filed in Australia, on October 30, 2002, and 30 application No, 2003904813 filed in Australia on September 4, 2003, the content of which is incorporated hereinto by reference. 122 WO 2004/039814 PCT/JP2003/013684 wherein 6 R1 is lower alkyl, hydroxy(lower)alkyl or halo(lower)alkyl, and R2 is hydrogen or amino protecting group, or R1 and R2 are bonded together and form lower alkylene Or lower alkenylene; 10 R3 is hydrogen or lower alkyl; R4 is 15 wherein X is O or NH, R7 is hydrogen, lower alkyl or amino protecting group. R8 is hydrogen or hydroxy, protected amino, guanidino, protected guanidino or saturated 3-to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or 25 protected amino, k, m, n and q are independently 0 or 1, and 123 WO 2004/039814 PCT/JP2003/013684 p is 0, 1, 2 or 3; R5 is carboxy or protected carboxy; and R5 is amino or protected amino, or a pharmaceutically acceptable salt thereof. 5 2. The compound of claim 1 wherein R1 is lower alkyl or hydroxy (lower) alkyl , and R2 is hydrogen or amino protecting group, or R1 and R2 are bonded together and form lower alkylene; 10 R3 is hydrogen; A is wherein X is O or NH; R7 is hydrogen or amino protecting group; 15 R3 is amino or protected amino; and p is 0 , 1 or . or a pharmaceutically acceptable salt thereof. 3. Ths compound of claim 2 wherein R3 is hydrogen, or a 20 pharmaceutically acceptable salt thereof. 4. The compound of claim 1 wherein R1 is lower alkyl, hydroxy(lower)alky1 or halo (lower) alkyl, and 25 R2 is hydrogen, aryl(lower)alkyl or acyl, or R1 and R2are bonded together and form lower alkylene or lower alkenylene; R5 is carboxy or esterified carboxy; R6 is amino or acylamino; 30 R7 is hydrogen, lower alkyl or acyl; and R8 is amino, mono or di(lower)alky1 amino, acylamino, guanidino, acylguanidino or saturated 3- to 8- membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or 35 acylamino, or a pharmaceutically acceptable salt thereof. 124 WO 2004/039814 PCT/JP2003/013684 5. The compound of claim 4 wherein R1 is lower alkyl or hydroxy(lower)alkyl, and R2 is hydrogen, aryl(lower)alkyl or acyl, or R1 and R2 are bonded together and form lower alkylene; 5 R5 is carboxy or esterified carboxy; R6 is amino acylamino; R7 is hydrogen or acyl; and R8 is amino or acylamino. or a pharmaceutically acceptable salt therof. 10 6. The compound of claim 5 wherein R1 is alkyl or hydroxy (lower) alkyl, and R2 is hydrogen, aryl(lower)alkyl, lower alkanoyl or lower alkoxycarbonyl, or 15 R1 and R2 are bonded together and form lewer alkylene; R5 is carboxy or lower alkoxycarbonyl; R6 is amino lower alkanoylamino or lower alkoxycarbonylamino; R1 is hydrogen, lower alkanoyl or lower alkoxycarbonyl; 20 and R5 is amino, lower alkanoylamino or lower alkoxycarbonylamino, or a pharmaceutically acceptable salt thereof. 25 7. The compound of claim 6 wherein Rl is lower alkyl or hydroxy(lower)alkyl, and R2 is hydrogern, or R1 and R2 are bonded together and form lower alkylene; R5 is carboxy; 30 R6 is amino; R7 is hydrogen or lower alkanoyl; and R9 is amino, or a pharmaceutically acceptable salt thereof. 35 8. The compound of claim 1 wherein R4 is selected from the group consisting of —NH—A-f NH)—(CH2)—(CH2) p-R14 125 WO 2004/039814 PCT/JP2003/013684 5 wherein R7, A, m, p and q are each as defined in claim 1, R14 is amino, mono or di (lower)alkylamino or protected amino , R15 is guanidino or protected guanidino, and 10 R16 is saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or protected amino, or a pharmaceutically acceptable salt thereof. 15 9. The compound of claim 1 wherein R4 is selected from the group consisting of 126 WO 2004/039814 PCT/JP2003/013684 wherein 5 p is 0, 1 or 2, q is 0 or 1, R7 is hydrogen or amino protecting group, and R9 is amino or protected amino, or a pharmaceutically acceptable salt thereof. 10 10. The compound of claim 9 wherein R7 is hydrogen, lower alkanoyl or lower alkoxycarbonyl; and R9 is amino, lower alkanoylamino or lower 15 a1koxy carbonylamino , or a pharmaceutocally acceptable salt thereof. 11. The compound of claim 10 wherein R7 is hydrogen or lower alkanoyl; and 20 R9 is amino, or a pharmaceutically acceptable salt thereof. 12. A process for preparing a compound of the formula [I]: 127 WO 2004/039814 PCT/JP2003/013684 Wherein Rl is lower alkyl, hydroxy (lower) alkyl or halo(lower)alkyl, and R2 is hydrogen or amino protecting group, or 5 R1 and R2 are bonded together and form lower alkylene or lower alkenylene; R3 is hydrogen or lower alkyl; R4 is wherein is 0 or NH, R7 is hydrogen, lower alkyl or aminc protecting 15 group, R8 is hydrogen or hydroxy, R9 is amino, mono or di(lower)alkylamino , protected amino, guanidino, protected guanidino or saturated 3- to 8-membered 20 heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or protected amino, k, m, n and q are independently 0 or 1, and p is 0, 1, 2 or 3; 25 R5 is carboxy or protected carboxy; and R6 is amino or protected amino, or a salt thereof, which comprises (1) reacting a compound of the formula. [II] : 128 WO 2004/039814 PCT/JP2003/013684 wherein R1, R2, R3 and R4 are each as defined above, or its reactive derivative at the amino group, or a salt 5 thereof with a compound of the formula [III]: wherein R5 and R6 are each as defined above, or its reactive derivative at the carboxy group, or a salt thereof to give a compound of the formula [I] : wherein R1, R2, R3, R4, R5 and R6 are each as defined0 above, or a salt thereof, or (2) subjecting a compound of the formula [Ia]: 129 WO 2004/039814 PCT/JP2003/013684 wherein R1 , R2, R3, R4 , R5 , R6 , R7 ,R8 A, k, m, n, p and q are each as defined above, and R9a is protected amino, protected guanidino or saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitiogen atoms 5 substituted by protected amino, or a salt thereof to elimination reaction of the amino protecting group to give a compound of the formula [Ib] : 10 wherein R1 , R2, R3, R5 , R6, R7 , R8 , A, k, m, n, p and q are each as defined above, and R9b is amino, guanidino or saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms substituted by amino, or a salt thereof, or 15 (3) reacting a compound of the formula [VI] : wherein R5and R6 are each as defined above, R10 is protected carboxy, and Y is a leaving group, or a salt thereof with a compound of the formula [VII] : 20 wherein R1 , R2, R3 and R4 are each as defined above, or a salt thereof to give a compound of the formula [VIII]: 130 WO 2004/039814 PCT/JP2003/013684 whrein R1 , R2 , R3 , R4 , R5, R6 and R10 are each as defined above, and Z**** is an anion, or a salt thereof, and subjecting the compound of the formula [VIII] or a salt 5 thereof to elimination reaction of the carboxy protecting group, to give a compound of the formula [I]: wherein R1 , R2 , R3 , R4 R5 and R6 are each as defined above, or a salt thereof. 10 13. A pharmaceutical composition comprising a compound of claim 1 or a pharmaceutically acceptable salt thereof in admixture with a pharmaceutically acceptable carrier. 15 14. A compound of claim 1 on a pharmaceutically acceptable salt thereof for use as a medicament. 15 A compound of claim 1 or a pharmaceutically acceptable salt thereof for use as an antimicrobial 20 agent. 16. Use of a compound of claim 1 or a pharmaceutically acceptable salt thereof for manufacture of a medicament for treating infectious diseases, 131 WO 2004/039814 PCT/JP2003/013684 17. A method for the treatment of infectious diseases which comprising administering a compound of claim 1 or a pharmaceutically acceptable salt thereof to human or 5 animals. 132 WO 2004/039814 PCT/JP2003/013684 DESCRIPTION CEPHEM COMPOUNDS TECHNICAL FIELD The present invention relates to new cephem 5 compounds and pharmaceutically acceptable salts thereof. More particularly, the present, invention relates to new cephem compounds and pharmaceutically acceptable salts thereof, which have antimicrobial activities, to processes for preparation thereof, to pharmaceutical 10 composition comprising the same, and to a method for treating infectious diseases in human being and animals. DISCLOSURE OF INVENTION One object of the present invention is to provide novel cephem compounds and pharmaceutically acceptable 15 salts thereof, which are highly active against a number of pathogenic microorganisms. Another object of the present invention is to provide processes for the preparation of said cephem compounds and salts thereof. 20 A further object of ths present invention is to provide a pharmaceutical composition comprising, as an active ingredient, said cephem compounds or their pharmaceutically acceptable salts. Still further object of the present invention is 25 to provide a method for treating infectious diseases caused by pathogenic microorganisms, which comprises administering said cephem compounds to infect&d human being or animals. The object cephem compounds of the present 30 invention are novel and can be represented by the following general formula [I]: 1 WO 2004/039814 PCT/JP2003/013684 wherein R1 is lower alkyl, hydroxy (lower) alkyl or halo(lower)alkyl, and 5 R2 is hydrogen or amino protecting group, or R1 and R2are bonded together and form lower alkylene or lower alkenylene; R3 is hydrogen or lower alkyl; R1 is wherein X is O or NH, 15 R7 is hydrogen, lower alkyl or amino protecting group, R3 is hydrogen or hydroxy, R9 is amino, mono or di(lower)alkylamino, protected amino, guanidino, protected 30 guanidino or saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or protected amino, k, m, n and q are independently 0 or 1, and 25 p is 0, 1, 2 or 3; R5 is carboxy or protected carboxy, and 2 WO 2004/039814 CT/JP2003/013684 R6 is amino or protected amino. As to the object compound (I), the following points are to be noted. That is , the object compound (I) include syn 5 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: 10 wherein R5 and R6 are each as defined above, and anti isomer (E form) means the other geometrical isomer having the partial structure represented by the following formula: 15 wherein R5 and R6 are each as defined above, and all of such geometrical isomer and mixture thereof are included within the scope ox this invention. In the present specification and claims, the partial structure of these geometrical isomers and 20 mixture thereof are represented for convenience' sake by the following formula: wherein R5 and R6 are each as defined above. Another point to be noted is that the pyrazolio 3 WO 2004/039814 PCT/JP2003/013684 moiety of the compound [I] can also exist in the tautomeric form, and such tautomeric equilibrium can be represented by the following formula. 5 wherein R1,R2,R3andR4 are each as defined above. Both of the above tautomeric isomers are included within the scope of the present invention, and in the present specification, and claims, however, the object compound [I] is represented for convenience' sake by one 10 expression of the pyrazolio group of the formula (A) . The cephem compound [I] of the present invention can be prepared by the following processes as illustrated, in the following. 15 4 WO 2004/039814 PCT/JP2003/013684 Process 1 or its reactive or its reactive or its reactive or its reactive derivative at the derivative at the derivative at the derivative at the amino group, carboxy group, or a salt thereof or a salt thereof 5 WO 2004/039814 PCT/JP2003/013684 Process 2 5 6 WO 2004/039814 PCT/JP2003/013684 7 WO 2004/039814 PCT/JP2003/013684 wherein R1,R2,R3,R4,R5,R6,R7,R8, A, k, m, n, p and 5 q are each as defined above, R10 is protected carboxy. Y is a leaving group, Z**** is an anion, R1a is protected hydroxy(lower)alkyl, 10 R1b is hydroxy(lower) alkyl, R9a is protected amino, protected guanidino or saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms substituted by protected amino, and R9b is amino, guanidino or saturated 3-to8-membered 15 heterocyclic group containing 1 to 4 nitrogen atoms substituted by amino. 8 WO 2004/039814 PCT/JP2003/013684 The starting compounds [II] and [VI] can be prepared by the following processes. 5 9 WO 2004/039814 PCT/JP2003/013684 wherein R1, R2, R3, R4, R5, R6,R10,YandZ** are each as 5 defined above, R11 is protected amino, R12 is protected carboxy, and R13 is amino protecting group or lower alkyl. The starting cotopounds [VII] and [XI] or salts 10 thereof can be prepared by the methods disclosed in the Preparations 3—6, 8-47 and 49-102 describ&d later or similar manners thereto. In the above and subsequent descriptions of this specification, suitable examples of the various 15 definitions ate explained in detail as follows. The term "lower" is used to mean a group having 1 10 WO 2004/039814 PCT/JP2003/013684 to 6, preferably 1to 4,carbon atoms, unless otherwise indicated. Suitable "lower alkyl" and "lower alkyl1' moiety in "hydro(lower)alkyl" , "protected hydroxy (lower) alkyl" , 5 "aryl (lower)alkyl", "halo (lower)alkyl" and "mono or di(lower)alkyl amino", include straight or branched alkyl having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert- butyl, pentyl, tart-pentyl and hexyl, In which more 10 preferred one is C1-C4 alkyl. Suitable "hydroxy(lower)alkyl" includes hydroxy (C1-C5) alkyl such as hydroxymethyl, 1- hydroxy ethyl, 2-hydroxyethyl,, 1-hydroxypropyl, 2- hydrdxypropyl, 3-hydroxypropyl, 4—hydroxybutyl, 5- 15 hydroxypentyl and 6 —hydroxyhexyl, in which more preferred one is hydroxy (C1-C4) 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 20 and fluorine. Preferred examples of "halo(lower)alkyl" include fluoromethyl, difluromethyl, trifluoromethyl, chloromethyl, bromomethyl, 2—fluoroethyl, 2 ,2- difluoroethyl, 2,2,2-trifluoroethyl, 2—chloroethyl, 2,2- dichloroethyl, 2,2,2—"trichloroethyl, 3—fluoropropyl and 25 2,2,3,3,3-pentafluoropropyl, in which more preferred one is halo (C1-C4) alkyl. Suitable "mono or di (lower)alkylamino" includes mono or di(C1-C6) alkylamino such as methylamino, dimethy1amino, ethylamino, diethylamino, N-ethyl—N— 30 methylamino, propylamino, butylamino and N-ethyl-N- propylamino, in which more preferred one is mono or di (C1-C4)alkyl amino. Suitable "lower alkylene" formed by R1 and R2 includes straight alkylene having 1 to 6, preferably 2 35 to 4 carbon atoms, such as methylene, ethylene, trilmethylene- and tetramethylene In which more preferred one is straight alkylene having 2 or 3 carbon atoms. Suitable "lower alkylene" formed by R1 and R2 11 WO 2004/039814 PCT/JP2003/013684 includes straight alkenylene having 2 to 6, preferably 2 to 4 carbon atoms, such as vinylene and propeoylene, in which more preferred one is straight alkylene having 2 or 3 carbon atoms. 5 Suitable "saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms" includes azetidinyl (e.g., l-azetidinyl and 3-azetidinyl), pyrrolidinyl (e.g., l-pyrro0lidinyl and 3-pyrrolidinyl), imidazolidinyl (e.g., 1-imidazolidinyl and 4- 10 imidazolidinyl), piperidinyl (e.g., 1-piperidinyl and 4- piperidinyl) and piperazinyl (e.g. , 1-piperazinyl) , in which, more preferred one is saturated 4- to 6-membered heterocyclic group containing 1 to 4 nitrogen atoms. The saturated 3-to 8-membered heterocyclic group 15 containing 1 to 4 nitrogen atoms is optionally substituted by amino or protected amino. Suitable examples of "saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substitutted by amino or protected amino" include 1- 20 azetidinyl, 3-amino—l-azetidinyl, 3-tert- butoxycarbonylamino-l-azetidinyl, 3-azetidinyl, 1- pyrrolidinyl, 3-amino-l-pyrrolidinyl, 3-tert- butoxycarbonylamino-l-pyrrolidinyl, 3-pyrrolidinyl, 1— piperidinyl, 4-piperidinyl and 1-piperazinyl. 25 Suitable "aryl" moiety in "aryl (lower) alkyl" includes C6-C12 aryl such as phenyl and naphthyl, in which, more preferred one is phenyl. Suitable "aryl(lower)alky1" includes mono-, di- or triphenyl (lower) alkyl such as benzyl, phenethyl, 30 benzhydryl and trityl. Suitable "lower alkanoyl" and "lower alkanoyl" moiety in "lower alkanoylamino" include straight or branched alkanoyl having 1 to 6 carbon atoms, such as formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, 35 isovaleryl, pivaloyl and hexanoyl, in which more preferred one is C1-C4 alkanoyl. Suitable "lower alkoxy" moiety in "lower alkoxycarbonyl" and "lower alkoxycarbonylamino" includes 12 WO 2004/039814 PCT/JP2003/013684 straight or branched alkoxy having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentyloxy, tert- pentyloxy and hexyloxy, in which more preferred one is 5 C1-C4 alkoxy. Suitable "amino protecting group" in "protected amino" includes an acyl group as mentioned below, substituted or unsubstituted aryl(lower)alkylidene [e.g., benzylidene, hydroxybenzylidene, etc.], aryl(lower)alkyl 10 such as mono-, di—or tripaenyl (lower) alkyl) [e.g., benzyl, phenethyl, benzhydryl, trityl, etc.], and the like . Suitable "acyl" includes lower alkanoyl [e.g., formyl, acetyl, propionyl, hexanoyl, pivaloyl, etc.], 15 mono(or di or tri) halo (lower)alkanoyl [e.g-, chloroacetyl, trifluoroacetyl, etc.], lower alkoxycarbonyl [e.g., methoxycarbonyl, ethoxycarbonyl. tert-butoxycarbonyl, tert-pentyloxycarbonyl, hexyloxycarbonyl, etc.], carbamoyl, aroyl [e.g., benzoyl, 20 toluoyl, naphthoyl, etc, ] , aryl (lower) alkanoyl [e.g., phenylacetyl, phenylpropionyl, etc], aryloxycarbonyl (e.g., phenoxycarbonyl, naphthyloxycarbonyl, etc.], aryloxy (lower) alkanoyl [e.g., phenoxyacetyl, phenoxypropionyl, etc.], arylglyoxyloyl [e.g., 25 phenylglyoxyloyl, naphthylglyoxyloyl, etc.], aryl (lower) alkoxycarbonyl which optionally substituted by suitable substituent(s) [e.g., benzyloxycarbonyl, phenethyloxycarbonyl, p-nitrobenzyloxyearbonyl, etc.], and the like. 30 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 (C1-C6) alkyl, C1-C6 alkanoyl 35 and (C1-C6) alkoxycarbonyl. Preferable examples of "protected amino" include aryl(lower)alkylamino and acylamino, in which more preferred ones are aryl(lower)alkylamino, lower 13 WO 2004/039814 PCT/JP2003/013684 alkanoylamino and lower alkoxycarbonylamino, and particularly preferred ones are mono-, di- or triphenyl (C2-C5) alkylamino , C3-C5 alkanoylamino and (C2- C6)alkoxycarbonylamino. 5 Preferable examples of "protected guanidino" include acylguanidino (monoacylguanidino and diacylguanidino such as 2,3- bis [ (lower) alkoxycarbonyl]guanidino [e.g., 2 ,3-bis (tert- butoxycarbonyl) guanidinol , In which morepreferred one 10 is 2 , 3-bis [ (C1-C5) alkoxycarbonyl] gnanidino. 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 15 (e.g. , formyl. acetyl, propionyl,. hexanoyl, pivaloyl, etc.], mono (or di tri) halo (lower) alkanoyl, [e.g., chloroace tyl, trifluoroacetyl, etc.], lower alkoxycarbonyl, [e.g., methoxycarbonyl, ethoxycarbonyl, tert-butoxycarboyl, tert-pentyloxycarbonyl , 20 hexyloxycarbonyl, etc.] , carbamoyl, and -the like. Suitable "aryl (lower) alkyl" moiety in the "aryl (lower)alkyloxy" includes mono-, di- or triphenyl (lower) alkyl (e.g, , benzyl, phemethyl, benzydryl, trityl , etc.], and the like. 25 Suitable "protected carboxy" includes esterified carboxy and "the like, and concrete examples of estetrified carbaoxy include lower alkoxycarbonyl (e.g. , methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, isobutoxycarbonyl, 30 tert-butoxycarbonyl, pentyloxycarbonyl , hexyloxycarbonyl, l-cyclopropylethoxycarbonyl, etc. ] which may have suitable substituent (s) , for example, lower alkanoyloxy(lower)alkoxycarbonyl [e.g., acetoxymethoxycarbonyl, propionyloxymethosycarbonyl, 35 butyryloxymethoxycarbonyl, valeryloxymethoxycarbonyl, pivaloyloxymethoxycarbonyl, 1-acetoxyethoxycarbonyl, 1- propionyloxyethoxycarbonyl, 2 -propionyloxy thoxycarbo ny1, hexanoyloxymethoxycarbonyl, etc.], lower 14 WO 2004/039814 PCT/JP2003/013684 alkanesulfonyl (lower) alkoxycarbonyl, [e.g., 2- mesylethoxycarbonyl, etc, ] or mono (or di or tri) halo (lower) alkoxycarbonyl [e.g., 2- iodoethoxycarbony1, 2,2,2-trichloroethoxycarbonyl, 5 etc.I; lower alkenyloxycarbonyl [e.g., vinyloxycarbonyl, allyloxycarbonyl, etc.]; lower alkynyloxycarbonyl [e.g-, ethynyloxycarbonyl, propynyloxycarbonyl, etc.]; aryl(lower)alkoxycarbonyl which may have suitable substituent(s) [e.g. , benzyloxycarbony1, 4- 10 methoxybenzyloxycarbonyl, 4—nitrobezyloxycarbonyl, phenethyloxycarbonyl, trityloxycarbonyl, benzhydryloxycarbonyl, bis (methoxyphenyl) methoxycarbonyl, 3 ,4-dimethoxybenzyloxycarbonyl, 4-hydroxy-3 , 5-di-tert- butylbenzyloxycarbonyl, etc.]; aryloxycarbonyl which may 15 have suitable substituent(s) [e.g., phenoxycarbonyl, 4- chlorophenoxycarbonyl, tolyoxycarbonyl, 4-tert- butylphenoxycarbonyl , xylyloxycarbonyl, mesityloxycarbony1, cumanyloxycarbonyl, etc.]; and the like. 20 Preferable examples of "protected carboxy" include lower alkoxycarbonyl and aryl (lower) alkoxycarbonyl which may have suitable substituent(s) , in which more preferred one is (C1-C6) alkoxycarbanyl. Suitable "leaving group" includes halogen, [e.g., 35 chlorine, bromine, iodine, etc.] or acyloxy such as arylsulfonyloxy [e.g., benzenesulfonyloxy, tosyloxy, etc.], lower alkylsulfonyloxy [e.g., mesyloxy, etc.], lower alkanoyloxy [e.g., acetyloxy, propionyloxy, etc.], and the like. 30 Suitable "anion" includes formate, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate, toluenesulfonate, chloride, bromide, iodide, sulfate, hydrogen sulfate, phosphate, and the like. 35 Suitable pharmaceutically acceptable salts of the object compound [I] 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 15 WO 2004/039814 PCT/JP2003/013684 example, an alkali metal salt [e.g., sodium salt, potassium salt, etc.], an alkaline earth metal salt, [e.g., calcium salt, magnesium salt, etc] , an ammonium salt; a salt with an organic base, for example, an 5 organic amine salt [e.g., trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N,N'—dibenzylethylenediamine salt, etc.]; an inorganic acid addition salt [e.g, 10 hydrochloride, hydrobromide, sulfate, hydrogen sulfate, phosphate, etc,]; an organic carboxylic ot sulfonic acid addition salt [e.g., formate, acetate, trifluoroacetate, maleate, tartrate, citrate, fumarate, methanesulfonate, benzenesulfonate, toluenesulfoaate, etc.]; and a salt 15 with a basic or acidic amino acid [e.g., arginine, aspartic acid, glutamic acid, etc.]. The preferred embodiments of the cepkem compound of the present invention represented by the general formula [I] are as follows. 20 (1) The compound of the formula [I] wherein R1 is lower alkyl or hydroxy (lower) alky1,and R2 is hydrogen or amino protecting group, or R1 and R2 are bonded together and form lower alkylene; R3 is hydrogen; 30 p is 0r,1 or 2, or a pharmaceutically acceptable salt thereof. 12) The compound of (1) above wherein R3 is hydrogen, or a pharmaceutically acceptable salt thereof. (3) The compound of the formula [I] wherein 35 R1 is lower alkyl, hydroxy (lower) alkey1 or halo(lower)alkyl, and R2 is hydrogen, aryl(lower)alkyl or acyl, or 16 WO 2004/039814 PCT/JP2003/013684 R1 and R2 are bended together and form lower alkylene or lower alkenylene; R5 is carboxy or esterified carboxy; R6 is amino or acylamino; 5 R7 is hydrogen, lower alkyl or acyl; and R9 is amino, mono or di (lower) alkylamino , acylamino, guanidino, acylguanidino or saturated 3- to 8- membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or 10 acylamino, or a pharmaceutically acceptable salt thereof. (4) The compound of (3) above wherein R1 is lower alkyl or hydroxy (lower) akkyl, and R2 is hydrogen, aryl(lower)alkyl or acyl, or 15 R1 and R2 are bonded together and form lower alkylene; R5 is carboxy or esterified carboxy; R6 is amino or acylamino; R7 is hydrogen or acyl; and R9 is amino or acylannino, 20 or a pharmdceutically acceptable salt thereof. (5) The compound of (4) above wherein R1 is lower alkyl or hydroxy (lower) alky1, and R2 is hydrogen, aryl(lower)alkyl, lower alkanoyl or lower alkoxycarbonyl, or 25 R1 and R2 are bonded together and form lower alkylene; R5 is carboxy or lower alkoxycarbonyl; R6 is amino, lower alkanoylamino or lower alkoxycarbonylamino; R7 is hydrogen, lower alkanoyl or lower alkoxycarbonyl; 30 and R3 is amino, lower alkanoylamino or lower alkoxycarbonylamino, or a pharmaceutically acceptable salt thereof. (6) The compound of (5) above wherein 35 R1 is C1—C6 alkyl or hydroxy (C1-C6) alkyl, and R2 is hydrogen, mono-, di- or triphenyl (C1-C6) alkyl, C1-C6 alkanoyl or (C1-C6) alkoxycarbonyl, or R1 and R2 are bonded together and form C1-C6 alkylene; 17 WO 2004/039814 PCT/JP2003/013684 R5 is carboxy or (C1-C6) alkoxycarbonyl ; R6 is amino, C1-C6 alkanoylamino or (C1-C6) alkoxycarbonylamino ; R7 is hydrogen, C1-C6 alkanoyl or (C1-C6) alkoxycarbonyl; 5 and R9 is amino, C1-C6 alkanoylamino or (C1-C6) alkoxycarbonylamino , or a pharmaceutically acceptable salt thereof. (7) The compound of (5) above wherein 10 R1 is lower alkyl or hydroxy(lower)alkyl , and R2 is hydrogen, or R1 and R2 are bonded together and form lower alkylene; R5 is carboxy; R6 ia amino; 15 R7 is hydrogen or lower alkanoyl; and R9 is amino, or a. pharmaceutically acceptable salt thereof. (8) The compound of (7) above wherein, R1 is C1-C6 alkyl or hydroxy (C1-C6) alkyl , and 20 R2 is hydrogen, or R1 and R2 are bonded together and form C1-C6 aIkylene; R5 is carboxy; R6 is amino; R7 is hydrogen or C2-C6 alkanoyl; and 25 R9 is amino, or a pharmacetically acceptable salt thereof, (9) The compound of the formula [I] wherein R4 is selected from the group consisting of 18 WO 2004/039814 PCT/JP2003/013684 wherein R7, A, m, p and q are each as defined above in 5 the formula [I], R14 is amino, mono or di(lower)alkylamino or protected amino, Rl5 is guanidino or protected guanidino, and R16 is saturated 3- to 8-membered heterocyclic group 10 containing 1 to 4 nitrogen atoms optionally substituted by amino or protected amino, or a pharmaceutically acceptably salt thereof. (10) The compound of the formula [I] wherein R4 is selected from the group consisting of 19 WO 2004/039814 PCT/JP2003/013684 wherein p is 0, I or 2, q is 0 or 1, 5 R7 is hydrogen or amino protecting group, and R9 is amino or protected amino, or a pharmaceutically acceptable salt thereof. (11) The compound of (10) above wherein R7 is hydrogen, lower alkanoyl or lower alKoxycarbonyl; 10 and R3 is amino, lower alkanoylamino or lower alkoxycarbanylamino, or a pharmaceutically acceptable salt thereof. (12) The compound of (II) above wherein 15 R7 is hydrogen. C1-C6 alkanoyl or (C1-C6) alkoxycarbonyl; and R9 is amino, C1-C6 alkanoylamino or (C1-C6) alkoxycarbonylamino, or a pharmaceutically acceptable salt thereof. 20 (l3) The compound of (11) above wherein R7 is hydrogen or lower alkanoyl; and R9 is amino, or a pharmaceutically acceptable salt thereof. (14) The compound of (13) above wherein 25 R7 is hydrogen or C1-C5 alkanoyl; and R9 is amino, or a pharmaceutically acceptable salt thereof. The processes for preparing the object compound of 30 the present invention are explained in detail in the following. Process 1 The compound [I] or a salt thereof can be prepared by reacting the compound [II] or its reactive derivative 35 at the amino group, or a salt thereof with the compound [III] or its reactive derivative at the carboxy group, 20 WO 2004/039814 PCT/JP2003/013684 or a salt thereof. Suitable reactive derivative at the amino group of the compound [II] includes Schiff's base type amino or its tautomeric euamifle type isomer formed by the 5 reaction of the compound [II] with a carbonyl compound sucn as aldehyde, xetone and the like; a silyl derivative formed by the reaction of the compound [II] with a silyl compound such as bis(trimethylsilyl)acetamide, 10 mono (trimethylsilyl) acetamide [e.g., N- (trimethylsilyl) acetamide] , bis (trimethylsilyl) urea and the like; a derivative formed by the reaction of the compound [II] with phosphorus trichloride or phosgene. Suitable salts of the compound [II] and its 15 reactive derivative can be referred to the ones as exemplified for the compound [1]. 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. 20 A suitable example of the reactive derivatives may be an acid chloride; an acid azide; a. milted acid anhydride with an acid such as substituted phosphoric acid [e.g., dialkylphosphoric acid, phenylphosphoric a.cid, diphenylphosphoric acid, dibenzylphosphoric acid, 25 halogenated phosphoric acid, etc.], dialkylphosphorous acid, sulfurous acid, thiosulfuric acid, sulfuric acid, alkanesulfonic acid [e.g, methanesulfonic acid, etc.], aliphatic carboxylic acid [e.g., acetic acid, propionic acid, butyric acid, isobutyric acid, pivalic acid, 30 pentanoic acid, isopentanoic acid, 2-ethylbutyric acid, trichloroacetic acid, etc.] and aromatic carboxylic acid [e.g., benzoic acid, etc,]; a symmetrical acid anhydride; an activated amide with imidazole, 4- substituted imidazole, dimethylpyrazole, triazole or 35 tetrazole; an activated ester [e.g., cyanomethyl ester, methoxymethyl ester, dimethyliminoimethyl (CH3)2N+=CH-] ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, 21 WO 2004/039814 PCT/JP2003/013684 pentachlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenyl thioester, p-nitrophenyl thioester, p—cresyl thioester, carboxymethyl thioester, pyranyl ester, pyridyl ester, piperidyl ester, 8- 5 quinolyl thioester, etc.]; or an ester with an N-hydroxy compound [e.g.,N,N-dimethylhydroxylamine, 1-hydroxy-2- (1H)-pyridone, N-hydroxysuccinimide, N- hydroxyphthalimide, N-hydroxy-lH-benzotriazole, etc. ] . These reactive derivatives can optionally be selected 10 from them according to the kind of the compound [III] to be used. Suitably salts of the compound [III] and its reactive derivative can be referred to the ones as exemplified for the compound [I]. 15 The reaction is usually carried out in a conventional solvent such as water, alcohol [e.g., methanol, ethanol, etc.], acetone , dioxane , acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N ,N-dimethylformaraide , 20 pyridine or any other organic solvent which does not adversely affect the reaction. Thsae 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 25 preferably carried out in the presence of a conventional condensing agent such as N,N' dicyclohexylcarbodiimide; N-cyclohexyl-N'-morpholinoethylcarbodimide; N— cyclohexyl-N'—(4-diethylaminocyclohexyl) carbodiimide; N,N'-diethylcarbodiimide; N,N'-diisopropylcarbodiimide; 30 N-ethyl-N'-(3—dimethylaminopropyl)carbodiimide; N,N'— carbonyl-bis- (2—methylimidazole); pentamethyleneketane- N-cyclohexylimine; diphenylketene--N-cycloh.Qxylimine; ethoxyacetylene; I-alkoxy-I-chloroethylene; trialkyl phosphite; ethyl polyphosphate; isopropyl polyphosphate; 35 phosphorus oxychloride (phosphoryl chloride) ; phosphorus trichloride: thionyl chloride; oxalyl chloride; lower alkyl haloformate [e.g., ethyl chloroformate, isopropyl chloroformate, etc.], triphenylphosphine; 2-ethyl-7- 22 WO 2004/039814 PCT/JP2003/013684 hydroxybenzisoxazolium salt; 2-ethyl-5- (m- sulfophenyl)isonxazolium hydroxide intramolecular salt; l-(p-chlorobenzenesulfonyloxy) -6-chloro-lH- benzotriazole; so-called Vilsmeier reagent prepared by 5 the reaction of N,N-dimethylformamide with thionyl 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 bass such, as an 10 alkali metal bicarbonate, tri(lower)alkylamine, pyridine, N- (lower)alkylmorpholine, N,N—di(lower)aIkylbenzylamine, and the like. The reaction temperature is not critical, and the reaction is usually carried out under cooling to warming. 15 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. 20 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. 25 Suitable base includes an inorganic base and an organic base such as an alkali metal [e.g., sodium, potassium, etc.], an alkaline earth metal [e.g., magnesium, calcium, etc.], the hydroxide or carbonate or hydrogencarbonate thereof, trialkylamine [e.g., 30 trimethylamine, triethylamine, etc.], picoline, 1,5- 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 [e.g., 35 formic acid, acetic acid, propionic acid, trichloroacetic acid, trifluoroacetic acid, etc.], and an inorganic acid [e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, hydrogen chloride, hydrogen bromide, 23 WO 2004/039814 PCT/JP2003/013684 etc.]. The elimination using Lewis acid such as trihaloacetic acid [e.g., trichloroacetic acid, trifluoroacetic acid, etc.], and the like is preferably 5 carried out in the presence of cation trapping agents [e.g., anisole, phenol, etc.]. The reaction is usually carried out in a solvent such, as water, alcohol [e.g. , methanol, ethanol, etc:.] , methylene chloride, tetrahydrofuran, a mixture thereof 10 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 cut under cooling to warming. 15 Process 3-(i) The compound [VIII] or a salt thereof can be prepared by reacting the compound [VII or a salt thereof with the compound [VII] or a salt thereof. Suitable salt of the Compounds [VI], [VII] and 20 [VIII] can be referred to the ones as exemplified for the compound [I] . The present reaction may be carried out in a solvent such as water, phosphate buffer, acetone, chloroform, acetonitrile nitrobenzene, methylene 25 chloride, ethylene chloride, formamide, N ,N- dimethylformamide, methanol, ethanol, diethy1 ether, tetrahydrofuran, dimethyl sulfoxide, or any other organic solvent which does not adversely affect the reaction, preferably in ones having strong polarities. 30 Among the solvent, hydrophilic solvents may be used in a mixture with water. When the compound [VII is liquid, it can also be used as a solvent. The reaction is preferably conducted in the presence of a base, for example,- an inorganic base such 35 as alkali metal hydroxide, alkali metal carbonate, alkali methyl hydrogencarbonate , an organic basa such as trialkylamine, and the like. The reaction temperature is not critical, and the 24 WO 2004/039814 PCT/JP2003/013684 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 [e,g., sodium iodide, potassium iodide, etc.], 5 alkali metal thiocyanate [e.g., sodium thiocyanate, potassium thiocyanate, etc.], and the like. Anion Z**** may be one derived from a leaving group Y, and it may be converted to other anion by a conventional method. 10 Process 3-(ii) The compound [I] or a salt thereof can be prepared by subjecting the compound [VIII] or a salt thereof to elimination reaction of the carboxy protecting group. Elimination reaction is carried out in similar 15 manner to the reaction in the aforementioned Process 2, and therefore the reagents to be used and reaction conditions (e.g., solvent, reaction temperature, etc.) can be referred to those of Process 2. Process 4 20 The compound [Id] or a salt thereof can be prepared by subjecting the compound [Ic] or a salt thereof to elimination reaction of the hydroxy protecting group. Suitable method of this elimination reaction 25 includes conventional one such as hydrolysis, reduction and the like, (i) For hydrolysis; The hydrolysis is preferably carried out in the presence of a base or an acid including Lewis acid. 30 Suitable base includes an inorganic base and an organic base such as an alkali metal [e.g., sodium, potassium, etc.], an alkaline earth metal [e.g., magnesium, calcium, etc.], the hydroxide or carbonate or hydrogencarbonate thereof, trialkylamine [e,g., 35 trimethylamine, triethylamine, etc.], picoline, 1,5— 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. 25 WO 2004/039814 PCT/JP2003/013684 Suitable acid includes an organic acid [e.g., formic acid, acetic acid, propionic acid, trichloroacetic acid, trifluoroacetic acid, stc], and an inorganic acid [e.g., hydrochloric acid, hydrobromic 5 acid, sulfuric acid, hydrogen chloride, hydrogen bromide, etc.]. The elimination using Lewis acid such as trihaloacetic acid [e.g., tri-chlroacetic acid, trifluoroacetic acid, etc] and the like is preferably 10 carried out in the presence of cation trapping agents [e.g., anisole, phenol, etc.]. The reaction is usually carried out in a solvent such as water, alcohol [e.g., methanol , ethanol, etc.] , methylene chloride, t^trahydrofuran, a mixture thereof 15 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. 20 (ii) For reduction: Reduction is carried out in a conventional manner, including chemical reduction and catalytic reduction. Suitable reducing reagents to be used in chemical reduction are a combination of a metal [e.g., tin, zinc, 25 iron, etc] or metallic compound [e.g. , chromium chloride, chromium acetate, etc.] and an organic acid or inorganic acid [e.g., formic acid, acetic acid, propionic acid, trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid, hydrobromic acid, etc.]. 30 Suitable catalysts to be used in catalytic reduction are conventional ones such as platinum catalysts [e.g. , platinum plate, spongy platinum, platinum black, colloidal platinum, platinum oxide, platinum wire, etc.], palladium catalysts [e.g., spongy 35 palladium, palladium black, palladium oxide, palladium on carbon, colloidal palladium, palladium on barium sulfate, palladium on barium carbonate, etc], nickel catalysts [e.g., reduced nickel, nickel oxide, Raney 26 WO 2004/039814 PCT/JP2003/013684 nickel, etc], cobalt catalysts [e.g., reduced cobalt, Raney cobalt, etc.], iron catalysts [e.g., reduced iron, Raney iron, etc.],copper catalysts [e.g., reduced copper, Raney copper, ullman copper, etc.] and the like. 5 The reduction is usually carried out in a conventional solvent which does not adversely influence the reaction such as water, methanol, ethanol, propanol, N,N-dimethylformamide or a mixture thereof. Additionally, in case that the above—mentioned 10 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 reduction may be the above-mentioned solvent, and other conventional solvent such as diethyl ether, 15 dioxane, 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. When R6 is protected amino, the amino protecting 20 group in R6 can be eliminated by a conventional method such as hydrolysis. Processes A and B for the preparation of the starting compounds are explained in detail in the following. 25 Process A-(i) The compormd [XII] or a salt thereof can be prepared by reacting the compound [X] or a salt thereof with the compound [XI] or a salt thereof. This reaction, can be carried out in a similar 30 manner to the reaction in the aforementioned Process 3— (i), and therefore the reagents to be used and reaction conditions [e.g., solvent, reaction temperature, etc.) can be referred to those of Process 3- (i) . Process A-(ii) 35 The compound [II] or a salt thereof can be prepared by subjecting the compound [XII] or a salt thereof to elimination reaction of the amino protecting groups in R11 and R13 and the carboxy protecting group in 27 WO 2004/039814 PCT/JP2003/013684 R12 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 5 conditions (e.g., solvent, reaction temperature, etc.) can be referred to those of Process 2. Process B The compound [VI] or a salt thereof can be prepared by reacting the compound [XIII) or its reactive 10 derivative at the amino group, or a salt thereof with the compound [XIV] 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, 15 and therefore the reagents to be used and reaction conditions (e.g., solvent, reaction temperature, etc.) can be referred to those of Process_l. The compounds obtained by the above processes can be isolated and purified by a conventional method such 20 as pulverization, recrystallization, column chromatography, reprecipitation, and the like. It is to be noted that the compound [I] and other compounds may include one or more stereoisomer(a) such as optical isomer(s) and geometrical isomer(s) due to 25 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 object compounds [I] and pharmaceutically acceptable salts thereof include solvates [e.g., 30 enclosure compounds (e.g., hydrate, etc.)]. The object compound [I] and pharmaceutically acceptable salts thereof are novel and exhibit high antimicrobial activity, inhibiting the growth of a wide variety of pathogenic microorganisms including Gram— 35 positive and Gram-negative microorganisms and are useful as antimicrobial agents. Now in order to show the utility of the object compound [I] the test data on MIC (minimal inhibitory 28 WO 2004/039814 PCT/JP2003/013684 concentration) of a representative compound of this invention are shown in the following. Test method In vitro antibacterial activity was determined by 5 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 viabls cells per ml) was streaked on heart infusion agar (Hl-agar) containing 10 graded concentrations of representative -test compound, and tine minimal inhibitory concentration (MIC) was expressed in mg/ml after incubation at 37oC for 20 hours . Test compound Compound (a): 7- [ (Z) -2- (5-amino-l , 2 , 4-thiadiazol-3-yl) - 15 2- (l-carboxy-l-methylethoxyimino)acetamido]-3-[7-(3- aminopropionamido)-2,3-dihydro-5-(1H-imidazo[1,2- blpyrazolio)]methyl-3-cephem-4-carboxylate (Example 3) Compound (b): 7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)- 2- (l-carboxy-l-methylethoxyimino) acetamido] -3- [3-amino- 20 4- (3-aminopropionamido) -2-methyl-l-pyrazolio]methyl-3- cephem-4-carboxylate (Example 4) Compound (c): 7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)- 2—(1-carboxy-l-methylethoxyimino)acetamido]-3-[3-amino- 4— (aminoacetyl) amino-2-methyl-1- pyrazolio]methyl-3- 25 cephem-4-carboxylic acid hydrogen sulfate (Example 6) Compound (d) : 7- [(Z)-2-(5-amino-1,2,4-thiadiazol-3-yl)- 2—(1-carboxy-l-methylethoxyimino)acetamido]—3-(3-amino- 4-[3-(2-aminoethyl)ureido]-2-methyl-l-pyrazolio methyl- 3—cephem-4-carboxylic acid hydrogen sulfate (Example 7) 30 Compound (e): 7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-y1)- 2- (1-carboxy-l-methylethoxyimino) acetamido] -3- (3-amino- 4—guanidino-2—methyl-l-pyrazolio)methyl-3-cephem-4- carboxylic acid hydrogen sulfate (Example 11) Ceftazidime 35 29 WO 2004/039814 PCT/JP2003/013684 Test: results For therapeutic administration, the object 5 compound [I] 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 10 or inorganic solid or liquid excipient which is suitable for oral, parentetal 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 15 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, 20 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 [I] may vary from and also depend upon the age, conditions of the patient, 25 a kind of diseases, a kind of the compound (I) to be applied, etc. In general amounts between 1 mg and 4,0 00 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 mg of the object compounds [I] 30 of the present invention may be used in treating diseases infected by pathogenic microorganisms. The following Preparations and Examples are given 30 WO 2004/039814 PCT/JP2003/013684 for the purpose of illustrating the present invention in more detail. Preparation 1 To a solution of (Z) -2- (5-amino-l,2,4-thiadiazol- 5 3-y1) [(2-tert-butoxy-1,1-dimethy1-2- oxoethoxy) imino] ethanoic acid (5g) in a mixture of tetrahydrofuran (80 ml) and N,N-dimethylformaide (20 ml) was added a solution of sodium bis(trimethylsilyl)amide (8.33 g) in tetrahydrofuran (12 10 ml) , and the mixture was stirred for 15 minutes. To the reaction mixture was added a solution of di-tert-butyl dicarbonate (3.3 g) in tetrahydrofuran (20 ml) under ice-cooling, and the mixture was stirred under ice- cooling for 3 hours. To the reaction mixture was added 15 ethyl acetate, and the mixture was washed with 10% aqueous potassium hydrogen sulfate solution, and then washed with, a phosphate buffer (pH 6.86). The organic layer was separated, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The 20 residue was triturated with diisopropyl ether and dried in vacuo to give (Z)-2-(5—[(tert-butoxycarbonyl)amino]- 1,2,4-thiadiazol-3-Yl)[(2-tert-butoxy-l, l-dimethyl-2- oxoethoxy) imino] etlianoic acid (3.10 g) , IR(KBr) 3191.6. 2981.4, 1714.4, 1550.5, 1153.2, 1000.9 25 cm-1 1H-NMR(DMSO-d6) .37 (9H, s), 1.45 (6H,s), 1.50 (9H, S) , 12.7 (1H,s) ESI-MS: m/z=429(M-H) Preparation 2 30 A mixture of N ,N-dimethylformamide (0.640 ml) and phosphoryl chloride (0.781 ml ) was stirred at room temperature for 30 minutes . To the mixture were added tetrahydrofuran (4 ml) and (Z)-2-(5-[(tert- butoxycarboxyl)amino]-l,2,4-thiadiazol-3-yl [ (2-tert- 35 butoxy-1,1 -dimethyl—2-oxoethoxy) imino] ethanoic acid (3 g) at 4°C, and the reaction mixture was stirred at room temperature for 1 hour. Meanwhile, a mixture of benzhydryl 7-amino-3-chloromethyl-3-cephem-4- 31 WO 2004/039814 PCT/JP2003/013684 carboxylate hydrochloride (3 g) and N- (trimethylsilyl)acetamide (8.72 g) in tetrahydrofuran (15 ml) was warmed to make a clear solution. The solution was then, cooled to -20°C and added to the 5 activated acid solution obtained above. The reaction mixture was stirred at a temper attire of —10°C to 0°C for 1 hour and poured into a mixture of ethyl acetate and water. The aqueous layer was separated, and the organic layer was washed with brine, dried over anhydrous 10 magnesium sulfate and filtered. The filtrate was concentrated in vacuo and purified by column chromatography on silica gel eluting with hexane/ethyl acetate (3:2) to give benzhydryl 7(Z)-2-(5-tert- butoxycarboxYlamino—1 ,2, 4-thiadiazol-3—yl) -2- (1-tert- 15 butoxycarbonyl-1-methylethoxylmino) acetamido] -3- chloromethyl-3-cephem-4-carboxylate (4.79 g) . rR(KBr 2981.4, 1793.5, 1720.2, 1524.8, 1371.1, 1247.7, 1151.3 cm-1 1H-NMR(DMSO-d6) 1.39 (6H, s), 1-48 (3H, s), 1.5 0 (6H, 20 s), 3.58 (1H, d, J-l8.3Hz), 3.76 (1H, d, J=l8.3Hz), 4.44 (2H, s), 5.29 (1H, d, J-5.0Hz), 6.01 (lH, dd, J=8.6, 5.OHz), 6.97 (1H, s) , 7.2-7.6 (10H, m) , 9.65 (1H,d, J=5.OHz), 12.7 (1H, s) ESI-MS: m/z-849(M+Na) 25 preparation 3 To a solution of 5-amino-l-methylpyrazole (5 g) in ethanol (5O ml) was added isoamyl nitrite (6.92 ml) , and then 20% hydrochloric acid (5 drops) was added at 4°C. The reaction mixture was refluxed for 2 hours and cooled 30 to room temperature . To the reaction mixture was added diisopxopyl 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-nitirosopyrazole (3.53 g) , 35 1H-NMR(DMSO-d6) 3.51 (3H, s), 8.07 (2H, brs) , 8.51 (1H, s) APCI-MS; m/z=l27(M+H) Preparation 4 32 WO 2004/039814 PCT/JP2003/013684 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 5 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 sulfuric acid salt 10 (1. 71 g) . 1H-NMR(DMSO-d6) 3.54 (3H, s), 7.19 (1H, s) ESI-M:- m/z=113 (M+H) Preparation 5 To a suspension of 1,1'-carbonyldiimidazole (9.73 15 g) in dehydrated chloroform (72 ml) was added tert-butyl N-(2=aminoethyl)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 (l4.22 g) and 4,5-diamino-l- 20 methylpyrazole sulfuric acid salt (10.51 g) , and the mixture was stirred at 50oC for 15 hours. The insoluble materials were removed by filtration. To the filtrate were added chloroform (200 ml) and 5% aqueous sodium hydrogen carbonate solution (100 ml). The organic layer 25 was separated, and the aqueous layer was extracted with a mixed solvent of chloroform and methanol(4:1). The organic layers were combined, dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with ethyl acetate and dried 30 in vacuo to give 5-amino-4- (3- (2- [(tert- butoxycarbonyl)amino]ethyl)ureido)-1—methylpyrazole (14.0 g) as a solid. 1H-NMR(DMSO-d6) 1.38 (9H, s), 2.36-2.98 (2H, m) , 3.03- 3.07 (2H, m), 3.50 (3H, s), 4.81 (2H, br), 5.92 (lH, br), 35 6.80 (1H, br) , 6.96 (1H, s), 7.18 (1H, br) Example 1 To a solution of benzhydryl 7b- [ (Z)-2- (5-tert- butoxycarbonylamino-1,2,4-thiadiazol-3-yl) -2-(l-tert- 33 WO 2004/039814 PCT/JP2003/013684 butoxycarbonyl-l-methylethoxyimino) acetamido] -3- chloromethyl-3-cephem-4-carboxylate (500 mg) in N,N- dimethylformamide (1.0 ml) was added sodium iodide (10 0 mg) , and the mixture was stirred at room temperature for 5 3 0 minutes. To the reaction mixture was added a Solution of 5-amino-4—(3—(2—[(tert- butoxycarbonyl)amino]ethyl]ureido)-1-methylpyrazole (216 mg) in N,N-dimethylformamide (1.0 ml) . The whole mixture was stirred at 32oC for 4 hours. To the 10 resulting reaction mixture were added ethtyl acetate (50 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed with 10% acqueous sodium trifluoroacetate solution and brine, dried over anhydrous sodium, sulfate and filtered. The filtrate was 25 concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (75 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene chloride (1.8 ml) were added anisole (0.6 ml) 20 and trifluoroacetic acid (1.2 ml). The resulting solution was stirred at room temperature for 4 hours and poured into diisopropyl ether 180 ml) . The resulting precipitate was collected by filtration and dried in vacuo to give a crude product (380 mg), which was 25 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 with concentrated hydrochloric acid and 30 chromatographed on Diaion® 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 7— [ (Z) — 2— (5—amino— 1, 2 , 4—thiadiazol- 3-yl)—2-(1—carboxy-1-methylethoxyimino)acetamido]-3-(3- 35 amino-4-[3-(2-aminoethyl)ureido]-2-methyl-l- pyrazolio)methyl-3-cephem—4-carboxylate (21 mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s), 3.12 (2H, t, 34 WO 2004/039814 PCT/JP2003/013684 J=5.7HZ), 3.22 (1H, d, J=17.9Hz), 3.49 (1H, d, J-17.9Hz), 3.46 (2H, t, J=5.7Hz), 3.71 (3H, s), 4.95 (1H, d, J-15.6Hz) , 5.15 (1H, d, J=l5.6Hz), 5.25 (1H, d, J=4.6Hz), 5.84 (1H, d, J=4.6Hz), 7.69 (1H, s) 5 Preparation 6 To a solution of 5-amino-4-(3-[2-[(tert- butoxycaxbonyl] amino] ethyl)urido) -l-methylpyrazole (597 mg) and triethylamine (243 mg) in methylene chloride (10 ml) was added triphenylmethyl chloride (669 mg) , and the 10 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, 15 filtered and cocentrated in vacua. The residue was triturated with ethyl acetate to give 4-(3-(2-f(tert- butoxycarbonyl amino] ethyl)) l-methyl-5- triphenylmethylaminopyrazole (640 mg) as a solid. 1H-NMR(DMSO-d6) 1.38 (9H, s), 2.70 (3H, s), 2.94-2.96 20 (2H, m) , 2.99-3.01 (2H, m) , 5.68 (1H, brs) , 5.96 (lH, br) , 6.78 (1H, br) , 6.85 (H, br) , 7.00 (1H, s), 7.13- 7.15 (6H, m), 7.24-7.28 (9H, m) Preparation 7 To a solution of benzhydryl 7- [ (Z) -2- (5-amino- 25 l,2,4-thiadiazol-3-yl)-2- (l-tert-butoxycarbonyl-l- methylethoxyimino) acetamido)-3-chloromethyl-3-cephem-4- carboxylate (60 g) in toluene (600 ml) were added a solution of sodium iodide (61.8 g) in 0.05 mol phosphate buffer (pH 7, 50 0 ml) and tricaprylylmethylammonium 30 chloride (6.67 g). The mixture was stirred at room temperature for 15 hours . The reaction mixture was added to a mixture of ethyl acetate and water. The organic layer was washed with water and brine, and then dried over magnesium sulfate. The magnesium sulfate was 35 filtered off, and the filtrate was evaporated to 255 g under reduced pressure. The concentrate was poured into diisopropyl ether (2 L) . The resulting precipitate was collected by filtration and dried to give benzydryl 7b- 35 WO 2004/039814 PCT/JP2003/013684 [[Z)-2-(5-amino-l,2,4-thiadiazol-3-yl) -2-(1-tert- butoxycarbonyl—1-methylethoxyimino) acetamido] -3- iodomethyl-3-cephem-4-carboxYlai:e (59.4 g) , 1H-NMR(DMSO-d6) 1.39 (9H, s), 1.46 (6H, s), 3.57 and 5 3.87 (2H, ABq, J=l8.OHz), 3.76 (3H, S), 4.30 (2H, bs), 5.25 (1H, d, J=4.9Hz), 5.94 flH, dd, J=4.9, 8.7Hz), 6,95 (1H, bs) , 7.15-7.60 (10H, m) , 8.17 (2H, bs) , 9.53 (1H, d, J=8.7Hz) Example 2 10 To a solution of benzhydryl 7 [ (Z) -2- (5-amino- 1,2,4-thiadiazol-3-yl)-2-(l-tert-butoxycarbonyl-l- methylethoxyimino) acetamido] — 3-iodomethyl-3—cephem-4- carboxylate (810 mg) in N,N-dimethylformamide (2.4 ml) was added N— (trimethylsilyl) acetamide (656 mg) , and the 15 mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added a solution of 4-(3-(2- [(tert-butoxycarbonyl)amino]ethyl)ureido) -l-methyl-5- triphenylmethylaminopyrazole (640 mg) in mettylen chloride (1O ml). The whole mixture was stirred at room 20 temperature for 26 hours. To the resulting reaction mixture were added ethyl acetate (50 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed with 10% aqueous sodium trifluoroacetate solution and brine, dried over 25 anhydrous sodium sulfatet and filtered. The filtrate was concentrated to about: 5 ml vacuo. The concentrate was poured into diisopropyl ether (80 ml) , and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in 30 methylene chloride (2.380 ml) were added anisole (0.79 ml) and trifluoroacetic acid (1.58 ml) . The resulting solution was stirred at room temperature for 4 hours and poured into diisopropyl ether (80 ml) . The resulting precipitate was collected by filtration and dried in 35 vacuo to give a crude product (635 mg), which was purified by preparative HPLC utilizing 0DS column. The eluate containing a desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to 36 WO 2004/039814 PCT/JP2003/013684 about pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2—propanol. The eluate was concentrated to about 30 ml in vacuo and 5 lyophilized to give 7- [ (Z) -2-(5-amino-l ,2,4-thiadiazol- 3-y1) -2- (1-carboxy-l -methylethoxyimino) acetamido ] —3 — (3- amino-4- [3- (2-aminoethyl) ureido] -2-methyl-l pyrazolio )methyl-3-cephem-4—carboxylate (54 mg) as an amorphous solid. 10 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s) , 3.12 (2H, t, J=5.7Hz), 3.22 (1H, d, J=17.9Hz), 3.49 (1H, d, J=17.9Hz), 3.45 (2H, t, J=5.7Hz), 3.71 (3H, s), 4.95 (1H, d, J=l5.6Hz) 5.15 (1H,d, J=15.6Hz), 5.25 (lH, d, J=4.6Hz), 5.84 (1H, d, J=4.6Hz), 7.89 (1H, s) 15 Preparation 8 To a solution of 2,3-dihydro-lH-imidazo[1,2- b)pyrazole (120 g) in sulfuric acid (500 ml) was added potassium nitrate (111 g) under ice-cooling. The mixture was stirred at room temperature for 48 hours. 20 The reaction mixture was added to ice (2.0 kg). The crystalline residue was collected by filtration and dried invacuo to give 7-nitro-2,3-dihydro-1H- imidazo[1,2-b]pyrazole (132 g) as a solid. 1H-NMR(DMSO-d6) 4.05-4.03 (2H, m) , 4.17-4.20 (2H, m), 25 7.82 (1H, s), 7.97 (1H, br) Preparation 9 A suspension of 7-nitro-2,3-dihydro-lH— imidazo [1,2-b] pyrazole (97 g) in a mixed solvent of sulfuric acid (34 ml) and water (2000 ml) was treated 30 with 10% palladium on carbon (10 g) under a hydrogen atmosphere at room temperature for 4 days. After the catalyst was filtered off, the filtrate was concentrated in vacuo. The residue was triturated with methanol and dried in vacuo to give 7-amin0-2,3-dihydro-lH- 35 imidazol [1,2-b]pyrasole sulfuiric acid salt (90 , 2 g) as a solid. 1H-NMR(DMSO-d6) 3.87-3.90 (2H, m0 , 4.07-4.10 (2H, m) , 7.28 (lH, s) 37 WO 2004/039814 PCT/JP2003/013684 Preparation_l_0 To a solution of 7-amino-2,3-dihydro-lH- imidazo[,2-b]pyraols sulfuric acid salt (22 g) and N-ethyldiisopropylamine (2.84 g) in methylene chloride 5 (70 ml) was added N- [3- (tert-butoxycarbonylamino) - propionyloxy]succinimide (3.15 g) . The mixture was stirred at room temperature for 4 Lours. The reaction mixture was washed with saturated aqueous sodium hydrogen carbonate solution, and the organic layer was 10 dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The oily residue was purified by calumn chromatography on silica gel eluting with 5% methanol/chloroform to give 7— [3- (tert- butoxycarboxylamino) propionyl ] amino-2 , 3-dihydro-lH- 15 imidazo[l,2-b]pyrazole (2.2 g) as a solid, 1H-NMR(CDCl3) 1.44 (9H, s), 2.52 (2H, t, J-6.0Hz), 3.36-3.47 (2H, m) , 3. 96 (2H, t, J-8,2H2), 4 .18 (2H, t) J=8.2Hz), 5.16 (1H, br), 7.16 (1H, s), 7.90 (lH, br) Example 3 20 7b-[ (Z) -2-(5-Amino-l,2,4-thiadiazol-3-yl) -2- (l- carboxy-l-methylethoxyimino) acetamido] -3- [ 7- [3- aminopropionamido) -2 , 3-dihydro-5- (1H-imidazo [1,2- b]pyrazolio ]methyl-3-cephem-4carboxylate The title compound was obtained from benzhydryl 25 7 b[(Z)-2-(5-tert-butoxycarbonylamino-1,2,4-thiadiazol- 3-yl) -2- (1-tert-butoxycarbany 1-1-methylethoxyimino) - acetamido]-3-chloromethyl-3-cepbem-4-carboxylate and 7— [3— (tert-butoxycarbonylamino) propionyl] amino-2 , 3- dihydro-lH-imidazo[l,2-b]pyrazole in the same manner as 30 in Example 1 as an amorphous solid. 1H-NMR(D2O) 1.51 (3H, s), 1.52 (3H, s), 2.33 (2H, t, J=6.4Hz), 3.26 (1H, d, J=17.9Hz), 3.53 (1H, d, J=17.9Hz), 3.31 (2H, t, J=6.4Hz), 4.15 (2H, t, J=8.7HZ), 4.33 (1H, q,J=8.7Hz), 4.42 (1H, q, J=8.7Hz), 4.95 (1H, d, 35 J=l.5Hz), 5.03 (1H, d, J=l5.lHz), 5.25 (1H, d, J=5.0Hz), 5.84 (1H, d, J=5.0Hz), 8 . 06 (1H, s) Preparation 11 tert-Butyl [2-(5-amino-l-methyl-lH-pyrazol-4- 38 WO 2004/039814 PCT/JP2003/013684 ylcatbamoyl) ethyl] carbonate The title compound was obtained from 4,5-diamino- 1-methylpyrazole sulfuric acid salt and N-[3-(tert- butoxycarbonylamino)propionyloxy]succinimide in the same 5 manner as in preparation 10. 1H-NMR(DMSO-d6) 1.3 8 (9H, s) , 2.35 (2H, t, J=7.1Hz), 3.l8 (2H, q, J=7.lHz), 3.50 (3H, s) , 4.90 (2H, s), 6.83 (1H, t, J=7.1Hz) , 7.14 (1H, s) , 9.06 (1H , s) AP-MS: m/z=283 10 Preparation 12 tert-Butyl (2-[1-methy1-5-(tritylamino)-1H- pyrazol-4—ylcarbamoyl] ethyl)carbamate The title compound was obtained from tert—butyl (2- (5-amino-l-methyl-lH-pyrazol-4-ylcarlbamoyl) thyl] - 15 carbamate in the same manner as in Preparation 6. 1H-NMR(DMSO-d6) 1.39 (9H, s) , 2. 08 (2h, t, J=7.1Hz), 2.71 (3H, s) , 3.04 (2H, q, J=7-lHz), 5.57 (1H, s) , 6.72, (1H, t, J=7.1Hz), 7.1-7.4 (16H, m) , 8.25 (1H, s) Example 4 20 7b[ (Z) -2- (5-Amino-l,2,4-thiadiazol-3-yl) -2-(l- carboxy-l-methylethoxyimino) acetamido] -3-[3-amino-4- (3- aminopropionamido) -methyl-l-pyrazolio]methyl—3-cephem- 4-carboxylate The title compound was obtained from benzhydryl 25 7 b [(Z) -2-(5-tert-butoxycarbonylamino-l,2,4-thiadiazol- 3-yl) -2- ( 1-tert-butoxycarbonyl-l-methylethoxyimino) - acetamidol-3-ch1oromethyl-3-cephem-4-carboxylate and tert-butyl (2-([l-methyl-5-(tritylamino)-lH-pyrazol-4- yLcarbamoyl]-ethyl] carbamate in the, same manner as in 30 Example 1. 1H-NMR(D2O) 1.53 (6H, s) , 2.89 (2H, t, J=6.5Hz), 3.20 and 3,47 (2H, ABq, J=18Hz), 3.34 (2H, t, J=6.5HZ), 3.75 (3H, 5), 4.99 and 5.21 (2H, ABq, J=16Hz), 5.25 (1H, d J=4.8HZ), 5.85 (1H, d, J=4.8Hz), 8.02 (1H, s) 35 ESI-MS: m/z=674 (m+Na) Preparation 13 To a solution of 1, 3-bis (tert-butoxycarbornyl) -2- (trifluoromethylsulfonyl)guanidine (22.3 g) in 39 WO 2004/039814 PCT/JP2003/013684 dichloromethane [100 ml] were added 4 , 5-diamino-l- methylpyrazole sulfuric acid salt (10 g) and triethylamine (33.2 ml) at 4°C, and the mixture was stirred at room temperature overnight. The reaction 5 mixture was poured into a mixture of ethyl acetate and water. The aqueous layer was separated, and the organic layer was washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated in vacuo. The concentrate was poured into 10 acetonitrile, and the resulting precipitate was collected by filtration and dried in vacuo to give 5- amino-4-[2' ,3'-bis(tert-butoxycarbonyl)quapidino1-1- methylpyrazole [11.62 g). 1H-NHR(DMSO-d6) 1.37 (9H, s), 1.50 (9H, s) , 3.52 (3H, 15 s) , 5.14 (2H, s) , 7.11 (1H, s) , 9.14 (IH, s), 3.1.5 (1H, s) ESl-MS: m/z=353 (M-H) Preparation 14 4- [2 ', 3 '-Bis (tert-butoxycarbonyl) quanidino] -1- 20 methyl-5- (tritylamino) pyrazole The title compound was obtained from 5-amino-4- [2 ' , 3 ' -bis (tert-butoxycarbonyl) guanidino] -1- methylpyrazole in the same manner as in Preparation 6, 1H-NMR(DMSO-d6) 1.37 (9H ,s) 1.50 (9H, s), 2.35 (3H, 25 s) , 5.88 (1H, s) , 7.17 (1H , s) . 7.21 (15H, m) , 8.85 (1H, a), 11.2 (1H, s) ESI-MS: m/z=597(M+H) Example 5 7 b- [(z) -2- [5-Amino-l, 2 , 4-thiadiazol-3-yl) -2- (1- 30 carboxy-1-methyletboxyimino)acetamido]-3-(3-amino-4- quanidino-2 -methy1-1 -pyrazo1io) methy 1- 3- cephem-4 - carboxylate The title compound was obtained from benzhydryl 7 b -[ (Z) -2- (5-amino-l,2,4-thiadiazol-3-yl) -2-(l-tert- 35 butoxycarbonyl-l-methylethoxyimino)acetamido]-3- iodomethyl-3-cephem-4-carboxylate and 4-[2 ' , 3'-bis ( tert- butoxycarbonyl) quanidino]-l-methyl-5- (tritylamino) pynazole in the same manner as in Example 1. 40 WO 2004/039814 PCT/JP2003/013684 1H-NMR (DMSO-d6) 1.53 (6H, s) , 3.25 and 3.57 (2H, ABq, J=l8Hz) , 3.75 (3H, S),0 5.00 and 5.18 (2H, ABq, J=l5Hz) , 5.21 (1H, d, J=4.9HZ) , 5.85 (1H, d, J=4.9Hz), 8.05 (1H, 5 Preparation 15 To a suspension of 4 , 5-diamino-l-methylpyrazole sulfuric acid 5 salt (305 g) in tetrahydrofuran (3.05 L) was added tert-butyl 2-[(2,5-dioxo-l-pyrrolidinyl)oxy]- 2-oxoethylcarbamate (415 g) under ice-cooling. To the 10 mixture was added diisopropylethylamine (556 ml) dropwise at a temperature below 10°C. The mixture was stirred at room temperature overnight. To the resulting solution were added brine and saturated aqueous sodium hydrogen carbonate solution, and the mixture was 15 extracted with ethyl acetate (3.0 L) . The aqueous layer was extracted with tetrahydrofuran/ethyl acetate=l/l (3.0 L) twice. The organic layer was dried over anhydrous macflie^ium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl 20 ether (1.0 L) and dried in vacuo to give tert-butyl 2- [ (5-amino-1 -methyl-1H-pyrazol-4-yl) amino ] -2- oxoethylcarbamate (307 g) . IR(Br) 3440, 3349, 1670, 1631, 1525, 1276, 1163, 1074, 1014, 860, 791 cm-1 25 1H-NMR(DHSO-d6) 1.39 (9H, s) , 3.44 (3H, s), 3.64 (2H, d, J=5.9Hz), 4.91 (2H, brs), 6.97 (1H, t, J=5.9Hz), 7.15 (1H, s) , 9.09 (1H, brs) Preparation 16 To a solution of tert-butyl 2- [ (5-amino-l-methyl- 30 lH-pyrazol-4-yl]amino] -2-oxoethylecarbamate (307 g) in N,N'-dimethylformamide (1.5 L) was added triphenylmethyl chloride (334 g) , To the mixture was added triethylamine (318 ml) dropwise. The mixture was stirred at room temperature for 1 hour. The reaction 35 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 41 WO 2004/039814 PCT/JP2003/013684 concentrated in vacuo. The residue was triturated with acetonitrile (1.5 L) and dried in vacuo to give tert- butyl 2-( l-methyl-5-(tritylamino)-lH-pyrazol—4- yl]amino]-2-oxoethylcarbamate (468 g) . 5 IR(KBr) 3336, 3280, 1724, 1683, 1599, 1234, 939, 704 cm-1 1H-NMR(DMSO-d6) 1.38 (9H, s) , 2.73 (3H, s), 3.38 (2H, d, J=5.8Hz), 5.58 (1H, s) , 6.94 (1H, t, J=5.8Hz), 7.11-7.35 (15H, m) , 7.21 (1H, s), 8.31 (1H, s) 10 ESI-MS: m/z=5l2.3(M+H+) Example 6 To a solution of benzhydryl 7—t(Z)-2-(5-amino- 1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1- metylethoxyimino) acetamido] — 3-chloromethyl-3—cephem—4- 15 carboxylate (489 g) in N,N-dimethylformamide (1.4 L) was added sodium iodide (102 g) . after stirring at room temperature for 1 hour, tert-butyl 2-{[l-methyl-5- (tritylamino) —lH-pyrazol-4-yl] amino)-2-oxoethylcarbamate (383 g) was added to the mixture. Stirring was 20 continued at 37oC for 24 hours. The resulting mixture was poured into water and extracted with, ethyl acetate. The organic layer was washed successively with water, 10% acqueous sodium thiosulfate solution, brine and 10% aqueous sodium trifluoroacetate solution, dried over 25 magnesium sulfate, filtered and evaporated in vacuo. The residue was dissolved in ethyl acetate (3.5 L), and the solution was dropwise added to diisopropyl ether (36 L) . The precipitate was collected by filtration. The filter cake was washed with diisopropyl ether and dried 30 in vacuo. The obtained solid (700 g) was dissolved in dichloromethane (1.4 L), and to the solution were added anisole (700 ml) and trifluoroacetic acid (2.1 L) successively. After stirring at room temperature for 4 35 hours, the reaction mixture was poured into diisopropyl ether (3 0 L) . The precipitate was collected by filtration. The obtained solid was washed with diisopropyl ether and dried in vacuo. The crude product 42 WO 2004/039814 PCT/JP2003/013684 was dissolved in water (3.5 L), and the pH of the solution was adjusted to 7.0 with 28% aqueous ammonia solution. The insoluble material was filtered off, and the pH of the filtrate was adjusted to 1 with 5 concentrated hydrochloric acid. The insoluble material was filtered off again. The filtrate was chromatographed on Diaion® HP- 20 eluting with 20% aqueous 2—propanol. The eluate was concentrated to about 3.0 L in vacuo, and 2.0M sulfuric acid (102 ml) 10 was added, to the concentrate. The mixture mas lyophilized to give the crude product. The crude product was purified by preparative HPLC (pH 7.0 phosphate buffer and acetonitrile), and the eluate containing a desired product was concentrated to 15 about 6 L in vacuo. The concentrate was adjusted to about pH 1 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 eluting with 20% aqueous 2-propanol. The eluate was concentrated to about 550 ml in vacuo, and 2.0M sulfuric acid (54.5 ml) 20 was added to the concentrate. To the mixture was added dropwise acetonitrile (880 ml), and the mixture was stirred at room temperature overnight. To the mixture was added acetonitrile (200 ml), and the mixture was stirred at room temperature for 2 hours. The resulting 25 white crystals were collected by filtration, washed with 25% aqueous acetonitrile and dried under reduced pressure to give 7[(Z)-2-(5-amino-l,2,4-thiadiazol-3— yl) —2— (l-carboxy-l-methylethoxyimino) acetamido] -3- [3- amino-4- (aminoacetyl)amino-2-methyl-l-pyrazoliolmethyl- 30 3-cephem-4-carboxylic acid hydrogen suIfate (72.5 g), IR(KBr) 1778, 1700, 1653, 1525, 1149, 1111, 617 cm-1 1H-NMR(D20] 1.61 (6H, s) , 3.22 and 3.45 (2H, ABq, J=l7.8Hz), 3.73 (3H, s), 4.03 (2H, s), 5.05 and 5.27 (2H, ABq, J=l5.8Hz), 5.25 (1H, d, J=4.8Hz), 5.87 (1H, d, 35 J=4.8az), 8.09 (1H, s) ESI-MS: m/z=638,2 t(M+H+) Example 7 A solution of 7-[(Z)-2-(5-amino-l,2,4-thiadizol- 43 WO 2004/039814 PCT/JP2003/013684 3-yl) -2- (1-carboxy-l-methylethoxyimino) acetamidol -3-(3- amino-4- [3- (2-aminoethyl) ureido] -2-methy1-1- pyrazolio)methyl—3-cephem-4-carboxylate (36 g) in water was purified by preparative HPLC utilizing ODS column. 5 The eluate containing a desired product was concentrated to about 1.5 L in vacuo. The concentrate was adjusted to about pH 1 with concentrated hydrochloric acid and chromatographed on Diaion® HP—20 (6 L) eluting with 20% aqueous 2-propanol. The eluate was concentrated to 10 about 800 ml in vacuo, and 2M sulfuric acid (17 ml) was added. The resulting solution was lyophilized to give a sulfuric acid salt as an amorphous powder (23.6 g). The powder was dissolved in water (71 ml) and ethanol (57 ml). After addition of seed crystals (310 15 mg) , which resulted in the precipitation of white solid, the mixture was stirred for 1 hour. A mixture of ethanol (47 ml) and water (37 ml) was added over 30 minutes, and ethanol (33 ml) was added over 2 0 minutes. Then the slurry was stirred for an additional 1.5 hour. 20 The precipitate was collected by filtration, washed with ethanol/water (60 ml/20 ml) and ethanol (60 ml) and dried to give 7-[(Z)-2- (5-amino-l,2,4-thiadiazol-3-yl)- 2- (1—carboxy-1—metnylethoxyimino) acetamido] -3 —[3—amino- 4— [3- (2—aminoethyl) ureido ]-2—methy1-1—pyrazolio] methyl- 25 3-cephem-4—carboxylic acid hydrogen sulfate as crystals (17.3 g). IR(KBr) 3353, 3183, 1778 , 1652, 1558 , 1403, 1321, 1143, 1118, 997, 619 cm-1 1H-NMR(D2O) 1.61 (6H, s), 3.10-3.55 (6H, m) , 3.71 (3H, 30 s), 5.02 and 5.23 (2H, ABq, J=16.7Hz), 5.25 (1H, d, J=4.9Hz), 5.87 (1H, d, J=4.9Hz), 7.91 (1H, s) ESI-MS: m/z=667 (M+H+) X-ray powder diffraction analysis (by Eigatu X-ray Diffraction system MultiFlex) 35 20 intensity 8.0 1286 12.7 586 13.8 423 44 WO 2004/039814 PCT/JP2003/013684 16.1 618 18.9 520 20.4 748 21.5 667 5 22.4 1058 23.3 944 24.0 618 25.5 813 26.7 472 10 279 537 28.5 455 31.3 390 X-ray: Cu/40 kV/30 mA Preparation 17 15 5-Amino-1- ethy1-4- nitrosopyrazole The title compound was obtained from 5-amino-l- ethylpyrazole in the same manner as in Preparation 3, 1H-NMR(DMSO-d6) d 1.21 (3H, t, J=7,lHz), 3.93 (2H, q, J=7.1Hz), 7.04 and 8. 53 (1H, s) , 8.10 and 8.15 (1H, brs) 20 APCI-MS: m/z=l41(M+H)+ Preparation 18 4,5-Diamino-l-ethylpyrazole sulfuric acid salt The title compound was obtained from 5-amino-l- ethyl-4-nitrosopyrasole in the same manner as in 25 Preparation 4. 1H-NMR(D2O) 1.36 (3H, t, J=-7.3Hz) , 4.10 (2H, q, J=7.3Hz), 7.77 (1H, s) ESI-MS: m/z=27 (M+H) + Preparation 19 30 5-Amino-4- [3- (tert-butoxycarbonylamino) - propionylamanol -l-ethylpyrazole The title compound was obtained from 4,5-diamino- 1—ethylpyrazole sulfuric acid salt in the same mannner as in Preparation 15. 35 1H-NMR(DMSO-d6) 1.24 (3H, t, J=7.2Hz), 1.37 (9H, s), 2.35 (2H, t, J=7.1HZ), 3.18 (2H, dt, J=7.1, 7.lHz), 3.35 (q, J=7.2Hz), 4.88 (2H, brs), 6.75-6.90 (1H, m), 7.17 (1H, s). 9.05 (1H, brs) 45 WO 2004/039814 PCT/JP2003/013684 APCI-MS: m/z=298(M+H)+ Preparation 20 4- [3- (tert-Butoxycarbonylamino)propionylamino] -1- ethyl-5—tripheaylmethylaminopyrazole 5 The title compound was obtained from 5-amino-4- [3- (tert-butoxycarbonylamino) propionylamino] —1- ethylpyrazole in the same manner as in Preparation 16. lH-NMR(DMSO-d6) 0.88 (3R, t, J=7.2Hz), 1.39 (9H, s) , 2.02 (2H ,t, J=7.1Hz), 2.95-3.20 (4H, m), 5.59 )(1H, brs), 10 6.60-6.75 (1H, m), 7.10-7.35 (16H, m), 8.04 (1H, brs) ESI-MS: m/z=540 (M+H)+, 562(M+Na)+ Example 8 7b[(Z)-2-(5-Amino-l,2,4-thiadiazol-3-yl)-2- (1- carboxy—1-methylethoxyimino) acetamido ) -3-[3-amino-4- (3- 15 aminopropionylamino) -2-ethyl-l-pyrazolio]me-tliyl-3 - cephem-4-carboxylate The title compound was obtained from benzhydryl 7 b [ (Z)-2- [5-amino-l,2,4-thiadiazol-3-yl)-2- (l-tert- butoxycarbonyl-1-methylethoxyimino) acetamido] -3- 20 iodomethyl—3-cephem-4-carboxylate and 4-[3-(tert- butoxycarbonylamino) propionylamino] -l-ethyl-5- triphenylmethylaminopyrazole in the same manner as in Example 1. IR(KBr) 3415, 1763, 1658, 1598, 1529, 1402, 1361 cm-1 25 1H-NMR(D2O) 1.33 (3H, t, J=7.2Hz), 1.53 (6H, s), 2.89 (2H, t, J=6.5Hz), 3.17 and 3.49 (2H, ABq, J=17.7Hz), 3.34 (2H, t, J=6.5HZ), 4,26 (2H, q, J=7.3Hz), 5.05 and 5.16 (2H, ABq, J=15.4H2), 5.26 (lH, dr J=4.8Hz), 5.85 (1H, d, J=4.8Hz), 8.03 (1H, s) 30 Preparation 21 tert-Butyl 2-[ (5-amino-l-ethyl-lH-pyrazol-4- yl) amino] -2-oxoethylcarbamate The title compound was obtained from 4,5-diamino- 1-ethylpyrazole sulftiric acid salt in the same manner as 35 in Preparation 15. 1H-NMR(DHSO-d6) 1.21 (3H, t, J=7.2Hz), 1.39 (9H, s), 3.64 (2H, d, J=6.0Hz), 3.86 (2H, d, J=7.2Hz), 4.88 (2H, brs), 6.90-7.00 (1H, m) , 7.17 (1H, s), 9.06 (1H, brs) 46 WO 2004/039814 PCT/JP2003/013684 ESI-MS: m/z=284 (M+H) + , 306(M+Na) + Preparation_2 2 tert-Butyl 2- {[l-eth.yl-5- (tritylamino) -1H-pyrazol- 4-yl] amino)-2-oxoethylcarbamate 5 The title compound was obtained from tert-butyl 2- [ (5-amino-1-ethyl-1H-pyrazol-4-yl) amino] -2- oxoethylcarbamate in the same manner as in Preparation 16. 1H-NMR(DMSO-d6) b 0.88 (3H, t, J=7.2Hz), 1.38 (9H, s), 10 3.16 (2H, q, J=7.2Hz), 3.31 (2H, d) , 5.59 (1H, brs) , 6.80-6.95 (1H, m), 7.10-7.40 (l6H, m), 8.03 (1H, brs) ESI-MS: m/z=526 (M+H)+, 548(M+Na}+ Example 9 7 b [ (Z) -2- (5Amino-1,2, 4-thiadiazol-3-yl) -2- (1- 15 carboxy-1 -methylethoxyimino) acetamido ] -3- [ 3 -amino-4- (aminoacetyl) amino-2-ethyl-l-pyrazolio] methyl-3-cephem- 4-carboxylate The title compound was obtained from bezhydryl 7 b [ (Z) -2-(5-amino-l, 2 , 4-thiadiazol-3-yl) -2- (l-tert- 20 butoxycarbony 1-1 -methylethoxyimino) acetamido] -3- iodomethyl -3-cephem-4-carboxylate and tert-butyl 2- [( 1- ethyl-5-(tritylamino) -lH-pyrazol-4-yl] amino } -2- oxoethylcarbamate in the same manner as in Example 1 - IR(KBR) 3444, 1761, 1635, 1626, 1445, 1406 cm-1 25 1H-NMR(D20) 1 . 33 [3H, t, J=7. 2Hz) , 1.53 (6H, s), 2.89 (2H, t, J=6.5Hz), 3.17 and 3.49 (2H, ABq, J=17.7Hz), 4.00 (2H, s), 4.28 [2H, q, J=7.2Hz), 5.06 and 5.17 (2H, ABq, J=15.4Hz) , 5.27 (1H, d, J=4.3Hz), 5.85 (1H, d, J=4. 8Hz) , 8.07 (lH, s) 30 Preparation 23 5-Amino-4-[2' ,3'-bis (tert-butoxycarbonyl)- quanidinol -l-ethylpyrazole The title compound was obtained from 1,3-bis(tert- butoxycarbonyl)-2- (trifluoromethylsulfonyl) quanidine and 35 4,5-diamino-l-ethylpyrazole sulfuric acid salt in the same manner as in Preparation 13. 1H-NMR(DMSO-d6) 1.22 (3H, t, J=7.1Hz), 1.37 (9H, s), 1.50 (9H, s) , 3.88 (2H, d, J=7.1Hz),, 5.12 (2H, brs) , 47 WO 2004/039814 PCT/JP2003/013684 7.14 (1H, s) , 9.16 (1H, brs) , 11.51 (1H, brs) ESI-MS: m/z=369(M+H)+ Preparation 24 4-[2' ,3 '-Bis(tert-butoxycarbonyl)guanidino]-l- 5 ethyl-5-triphenylmethylaminopyrazole The title compound was obtained from 5-amino- 4- [2',3'-bis(tert-butoxycarbonyl)quanidino]-1- ethylpyrazole in the same manner as in Preparation 16. 1H-NMR(DMSO-d6) 0.8 6 (3H, t, J=7.1Hz), 1.3 8 (9H, s), 10 1.49 (9H,s) , 5. 85 (1H, brs) , 7.10-7.30 (16H, m) , 8.80 (1H, brs), 11.14 (lHr, brs) ESI-MS: m/z = 611 (M+H) +, 633 (M+Na)+ Example 10 7 b -[(Z) -2-(5-Amino-l,2,4-thiadiazol-3-yl)-2- (1- 15 carboxy-1 -methylethoxyimino) acetamidol-3-[3-amino-2- ethyl-4-quanidino-l-pyra zolio]methyl-3-cephem-4- carboxylate The title compound was obtained from benzhydryl 7 b -Z) -2- (5-amino-1,2,4 -thiadiazol -3-yl) -2 - (1 -tert- 20 butoxycarbonyl-l-methylethoxyimino) acetamido] -3- iodomethyl-3-cephem-4-carbonylate and 4- [2 ' , 3 ' -bis ( tert- butoxycarbonyl) quanidino] -l-ethyl-5- triphenylmethylaminopyrazola in the same manner as in Example 1. 25 IR(KBr) 3437, 1760, 1658, 1625, 1406, 1065 cm-1 lH-NMR(D20) 1.35 (3H, t, J=7.3Hz), 1.53 (6H, a), 3.26 and 3.61 (2H, ABq, J=17.8Hz), 4.25 (2H, q , J=7.3Hz), 5.06 and 5.17 (2H, ABq, J=15.7Hz), 5.29 (1H, d, J=4 . 8Hz) , 5.85 (1H, d, J=4.8Hz), 8.06 (lH, s) 30 Example II To a suspension of benzhydryl 7b-[ (Z)-2-{5-amino- 1, 2 ,4-thiadiazol-3-yl) -2- (l-tert-butoxycarbonyl-l- methylethoxyimino) acetamido] -3-iodomethyl-3-cephem-4- carboxylate (5 00 g) in N,N—dimethylformamide (2,5 L) was 35 added 4-[2',3'-bis(tert-butoxycarbonyl)quanidino]-l- methyl-5-triphenylmethylaminopyrazole (419 g) and the mixture was stirred at room temperature for 16 hours. The reaction mixture was added to a mixture of ethyl 48 WO 2004/039814 PCT/JP2003/013684 acetate and water. The organic layer was washed with water, brine and 10% aqueous sodium trifluoroacetat solution and then dried over magnesium sulfate. The magnesium sulfate was filtered off, and the filtrate was 5 evaporated to 3.3 kg under reduced pressure. The concentrate was poured into diisopropyl ether (33 L), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene 10 chloride (1500 ml) were added anisole (500 ml) and trifluoroacetic acid (1500 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. The crude 15 product was dissolved in water (3.5 L), and the pH of the solution was adjusted to 7.0 with 28% aqueous ammonia solution. The insoluble material was filtared off, and the pH of the filtrate was adjusted to 1 with concentrated hydrochloric acid. The insoluble material 20 was filtered off, again. The filtrate was chromatographed on Diaion® HP-20 eluding with 20% aqueous 2-propanol, The eluate was concentrated to about 3.0 L in vacuo. To the concentrate was added 2.0M sulfuric acid (150 ml), and the mixture was lyophilized 25 to give the crude product. The crude product was purified with preparative HPLC utilising ODS column (pH 7.0 phosphate buffer and acetonitrile). The eluate containing a desired product was concentrated to about 6 1 in vacuo. The concentrate was adjusted to about pH 1 30 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 eluting with 20% aqueous 2-propanol. The eluate was concentrated to about 1.5 L in vacuo. To the concentrate was added 2.0M sulfuric acid (SO ml), and the mixture was lyophilized to give 7 b - [(Z) -2- (5- 35 amino-1,2,4-thiadiazol-3-yl)-2- (l-carboxy-1- methylethoxyimino) acetamido] -3-(3-amino-4—gnanidino-2- methyl-1-pyrazolio) methyl—3-cephem—4-carboxylic acid hydrogen sulfate (48.5g). 49 WO 2O041&398I4 PCT/JP30O3/013684 IR(KBr) 1776, 1714, 1577, 1651, 1402, 1112 cm-1 1H-NMR(D2O) 1.61 (6H, s) , 3.28 and 3,58 (2H, ABq, J=l7.8Hz), 3.74 (3H, s), 5.15 and 5.23 (2H, ABq, J=15.7Hz), 5.27 (1H, d, J=4.8Hz), 5.88 (1H, d, J=4.8Hz), 5 8.07 (1H, s) ESI-MS: m/z=623.2(M+H+) Preparation 25 To a suspension of 4 ,5-diamino-l(2- hydroxyethyl)pyrazole sulfuric acid salt (2.4 g) in 10 methylene chloride (40 ml) were added N- ethyldisopropylamine (2.1 ml) and N- [3- (tert- butoxycarbonyXamino)propionyloxy] succinimide (2.3 g) under ice-cooling, and the mixture was stirred at room temperature for 6 hours. To the reaction mixture were 15 added brine (40 ml) and saturated aqueous sodium hydrogen carbonate solution (20 ml), and the mixture was extracted with a mixture of ethyl acetate and 2-propanol (3:1, 60 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in 20 vacuo. The residue was triturated with diethyl ether to give 5-amino-4- [3- (tert-butoxycarbonylamino) propionyl] - amino-1-(2-hydroxyethyl)pyrazole (l.65 g) as a solid. 1H-NMR(DMSO-d6) 1.38 (9H, s) , 2.35 (2H, t, J=7.3Hz), 3.16-3.20 (2H, m), 3.62-3.65 (2H, m), 3.90 (2H, t, 25 J=6.0HZ), 4.85 (2H, brs) , 4.92 (1H, t, J=5.0HZ), 6.84 (1H, t, J=5.5Hz), 7.20 (1H, s), 9.09 (1H, brs) Example 12 7 b - [ (Z) -2- (5-Amino-l ,2 , 4-thiadiazol-3-yl) -2- (1- carboxy-l-methylethoxyimino)acetamido]-3-[3-amino-4-(3- 30 aminopropionamido)-2- (2-hydroxyethyl)-1- pyrazoliolmethy1-3-cephem-4-carboxylate The title compound was obtained from benzhydryl 7 b [ (Z) -2- (5-tert-butoxycarbonylamino-1, 2 , 4-thiadiazol- 3-yl)-1- (1-tert-butoxycarbonyl-l-methylethoxyimino)- 35 acetamido]-3-chloromethyl-3-cephem-4-carboxylate and 5- amino-4-[3- (tert-butoxycarbonylamino)propionyl] amino-1- [2-hydroxyethyl)pyrazole in the same manner as in Example 1 as an amorphous solid. 50 WO 2004/039814 PCT/JP2003/013684 1-H-NMR(D2O) 1.51 (6H, s) , 2.88 (2H, t, J=6 . 4Hz) , 3.15 (1H, d, J=17.9Hz), 3.48 (1H, d, J=17.9Hz), 3.32 [2H, t, J=6,4Hz), 3.88 (2H, t, J=4.3Hz), 4.39 (lH, dt, J=l6.5Hz, 4.8Hz), 4.42 (1H, dt, J=16.5, 4.8Hz), 5.06 (1H. d, 5 J=15.lHz), 5.11 (1H, d, J=15.lHz), 5.25 (lH, d, J=5.0Hz), 5.83 (lH, d, J=5.0Hz), 8.05 (1H, s) Preparation 26 To a solution of 4-formyl-4,5,6,7- tetrahydropyrazolo(1 .5-alpyrimidine (1.51 g) in sulfuric 10 acid (7.5 ml) was added potassium nitrate (111 g) under ice-cooling. The mixture was stirred at room temperature for 17 hours. The reaction mixture was added to ice (100 g) . The crystalline residue was collected by filtration and dried in vacuo to give 3- 15 nitro-4,5 ,6,7-tetralhydropyrazolo [1,5-alpyrimidine (0.63 g) as a solid. 1H-NMR(DMSO-d6) 2.00-2.05 (2H, m) , 3.30-3.36 (2H, m), 3.99 (2H, t, J=6.0Hz), 7.85 (1H, s), 7.89 (1H, s) Preparatjon27 20 A solution of 3-nitro-4,5,6,7- tetrahydropyrzolo [1,5-alpyrimidine (1.68 g) in a mixture of sulfuric acid (0.6 ml) , acetic acid (100 ml) and water (10 ml) was treated with 10% palladium on carbon (0.5 g) under a hydrogen atmosphere at room 25 temperature for 6 days. After the catalyst was filtered off, the filtrate was concentrated in vacao. The residue was triturated with ethanol and dried in vacuo to give 3-amino-4 ,5, 6 , 7-tetrahydropyrazolo [1 , 5- alpyrimidine sulfuric acid salt (2.3 g) as a solid. 30 1H-NMR (DMSO-d6) 1.97-2.01 (2H, m), 3.22 (2H, t, J=5.0Hz), 3.98 (2H, t, J=6.0Hz), 7.22 (1H, s) Preparation 28 To a solution of 3-amino-4,5,6,7- tetrahydropyrazolo[l,5-alpyrimidine sulfuric acid salt 35 (2.96 g) and N-ethyldiisopropylamine (3.88g) in methylene chloride (70 ml) was added 1,3-bis(tert- butoxycarbonyl)-2-(trifluoromethanesulfonyl) guanidine (3.91 g) . The mixture was stirred at room temperature 51 WO 2004/039814 PCT/JP2003/013684 for 150 minutes. The reaction mixture was washed with saturated aqueous sodium hydrogen, carbonate solution. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The 5 residue was purified by column chromatography on silica gel eluting with 2% methanol/chloroform to give 3-[2,3- bis (tert-butoxycarbonyl) guanidino ]-4,5 , 6 , 7-, tetrahydropyrazolo[l,5-alpyrimidine (3 . 4 g) as a solid. lH-NMR(CDCl3) 1.48 (9H, s), 1.52 (9H, s), 2.12-2.14 (2H, 10 m) , 3.33-3.37 (2H, m) , 4.08 (2H, t, J=6.0Hz), 6.17 (lH, brs), 7.16 (1H, s) , 9.87 (1H, brs), 11.39 (1H, brs) Example 13 To a solution of benzhydryl 7-[(Z)—2-[5—tert- butoxycarbonylamino-1,2,4-thiadiazol-3-yl)-2-fl-tert- 15 butoxycarbonyl-1-methylethoxyimino) acetamido] -3- chloromethyl-3-cephem-4-carboxylate (1.0 g) in ,N,N- dimethylformamide (2.0 ml) was added sodium iodide (181 mg) , and the mixture was stirred at room temperature for 30minutes. To the reaction mixture were added 3-[2,3- 20 bis(tert—butoxycarbonyl) guanidino]-4,5,6 ,7- tetrahydropyrazolo[1,5-a]pyrimidine (571 mg) and methylene chloride (2.0 ml). The whole mixture was stirred at room temperature for 7 hours. To the reaction mixture were added ethyl acetate (100 ml) and 25 water (50 ml) . The aqueous layer was separated, and the organic layer was washed with 10% aqueous sodium trifluoroacetate solution and brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into 30 diisopropyl ether (150 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 35 trifluoroacetic acid (2.0 ml), and the mixture was stirred at room temperature for 4 hours. The reaction mixture was poured into diisopropyl ether (150 ml), and the resulting precipitate was collected by filtration 52 WO 2004/039814 PCT/JP2003/013684 and driad in vacuo to give a crude product (570 mg) , 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 5 was adjusted to about pH 3 with concentrated hydrochloric acid and chromatographed on Diaionâ HP-20 [Mitsubishi Chemical Corporation] eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo and lyophlized to give 7-[(Z)-2- 10 (5-amino-1,2,4—thiadiaizol-3-yl) -2— (l-carboxy—1- methylethoxyimino) acetamido] -3- [3-guanidino-4,5,6,7- tetrahydro -l-pyrazolo [1 , 5-alpyrimidinio] methy 1- 3 - cephem- 4-carboylate (51mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s) , 1.53 (3H, s), 2.05-2.25 (2H, 15 m), 3.26 (1H, d, J=l7.4Hz), 3. 56 (lH, d, J=l7.4Hz], 3.30-3.45 (2H, m), 4.15 (2H, t, J=6.0Hz), 4.93 (1H, d, J=15.6Hz), 5.15 (1H, d, J=15.6Hz), 5.25 (1H, d, J=4.8Hz), 5.84 (1H, d, J=4.8Hz), 7.99 (lH, s) Preparation 29 20 To a solution of 7-amino-2 ,3-dihydro-lH- imidazol[1,2-b]pyrazole sulfuric acid salt (4.4 g) , 4- (dimethylamino) pyridine (244 mg) and triethylamine (8.10 g) in chloroform (45 ml) was added 1,3-bis(tert- butoxycarbonyl) -2- (trifluromethanesulfonyl) guanidine 25 (10.18 g), The mixture was stirred at room temperature for 2 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 30 magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diisopropyl ether to give 7- [2 , 3-bis (tert-butoxycarbonyl) guanidino] -2 , 3- dihydro-lH-imidazo [1,2,-b]pyrazole (4.6 g) as a solid. 1H-NMR(CDCl3) 1.49 (9H, s), 1.52 (9H, s) , 3.57-4.01 (2H, 35 m), 4.21 (2H, t, J=7.8Hz), 5.30 (1H, brs), 7.19 (1H, s), 9.86 (1H, brs), 11.32 (lH, brs) Example 14 7 b [(Z) -2-(5-Aminno-l,2,4-thiadiazol-3-yl)-2- (1- 53 WO 2004/039814 PCT/JP2003/013684 carboxy-l-methylethoxyimino) acetamido]-3- [7-guanidino- 2 ,3-dihydro-5- (lH-imidazo [1,2-b] pyrazolio) ]methyl-3- cephem- 4- carboxy1at e The title compound was obtained from benzhydryl 5 7 b - [ (Z) -2- (5-tert-butoxycarbonylamino-l ,2 ,4-thiadiazol- 3-yl) - 2-(l-tert-butoxycarbonyl-l-methylethoxyimino) - acetamido]-3-chloromethy1-3-cephem-4-carboxylate and 7 - [2 ,3-bis (tert-butoxycarbonyl) guanidino] -2, 3-diydro-lH- imidazo[1,2-b]pyrazole in the same manner as in Example 10 13 as an amorphous solid. 1H-NMR(D2O) 1.51 (3H, s), 1.52 (3H, s), 3.35 (1H, d, J=l7.9Hz), 3.61 (1H, d, J=17.9Hz), 4.19 (2H, t, J=8.7Hz), 4.37 (lH, q, J=8.7Hz), 4.47 (1H, q, J=8,7Hz), 5,00 (1H, d, J=15,lHz), 5.04 (1H, d, J=15.1HZ), 5. 26 (1H, d, 15 J=4.8Hz), 5.84 (1H, d, J=4.8HZ), 8.13 (1H, s) Praparation 30 To a salution of 5-amino-l— (2- hydroxyethyl)pyrazole (6.35 g) in a mixed solvent of ethanol (25 ml) and concentrated hydrochloric acid 20 [0.035 ml) was added dropwise isomyl nitrite (7.03 g) The mixture was stirred at room temperature for 17 hours. The crystalline residue was collected by filtration and dried in vacuo to give 5-amino-l-(2—hydroxyethyl)-4- nitrosopyrazole (4.0 g) as a solid. 25 1H-NMR(DMSO-d6) 3.68 (2H, t, J=5.5Hz), 3.94 (2H, t, J=5.5Hz], 4.89 (1H, br), 8.06(2H, br), 8.53 (1H, s) Preparation 31 A solution of 5-amino-l-(2-hydroxyethyl)-4- nitrosopyrazole (9 7 g) in a mixed solvent of sulfuric 30 acid (34 ml) and water (2000 ml) was treated with 10% paLLadium on carbon (10 g) under a hydtogen atmosphere at room temperature for 4 days, After the catalyst was filtered off, the filtrate was concentrated in vacuo. The residue was triturated with methanol and dried in 35 vacuo to give 4,5-diamino-l-(2-hydroxyethyl)pyrazole sulfuric acid salt (90.2 g) as a solid. lH-NMR(DMSO-d6) 3.66 (2H, t, J=5.5Hz), 3.9 5 (2H, t, J=5.5Hz), 7.25 (1H, s) 54 WO 2004/039814 PCT/JP2003/013684 Preparation_32 To a suspension of 4,5-diamino-l- (2- hydroxyethyllpyrazole sulfuric acid salt (50.0) g) in chloroform (500 ml) were added 4-(dimethyllamino) pyridine 5 (2.54 g) , triethylamine (116 ml) and 1,3-bis(tert- butoxycarbonyl)-2-(trifluoromethanesulfonyl) guanidine (106 g) • The mixture was stirred under reflux for 2 hours. After cooling on an ice bath, the reaction mixture was washed successively with water, 4% aqueous 10 nitric acid solution, water and aqueos sodium hydrogen carbanate solution. The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was triturated with a mixed solvent of ethyl acetate (50 ml) and diethyl ether (200 ml) to give 5- 15 amino-4- [ 2 , 3-bis( tert-butoxycarbonyl )guanidino]-l-(2- hydroxyethyl pyrazole (50 g) as a solid. 1H-NMR(CDCl3 ) 1.47 (9H, s) , 1.53 (9H,s) , 3.28 (1H, br) , 4.02-4.05 (4H, ml), 4.65 (2H, br) , 7,22 (1H, s) , 9,95 (1H, br), 11.55 (1H, br) 20 Example 15 7 [ (Z) -2-(5-Amino-l,2,4-thiadiazol-3-yl)-2- (1- carboxy-1- methylethoxyimino) acetamido]-3- [3-amino-4- guanidino-2- (2-hydroxyethyl) -l-pyrazolio]methyl-3- cephem- 4 - carboxy1ate 25 The title compound was obtained from benzhydryl 7 b - [ (Z) -2- (5-tert-butoxycarboxylamino-l, 2 , 4-thiadiazol- 3-yl) -2- (l-tert-butoxycarbonyl-l-methylethoxyimino) - acetamido]-3-chloromethyl-3-cephem-4-carboxylate and 5- amino-4- [2,3-bis (tert-butoxycarbonyl) guanidino] -1- (2- 30 hydroxyethyl)pyrazole in the same manner as in Example 13 as an amorphous solid. 1H-NMR(D2O) 1.52 (3H ,s), 3.21 (1H, d, J=l7.9Hz), 3.59 (1H, d, J=17.9Hz), 3.90 92H, t, J=4.8Hz), 4.35-4.50 (2H, m) , 5.07 (1H, d, J=14.9Hz), 5.11 (1H, d, J=14.9Hz), 5.28 35 (1H, d, J=5.0Hz), 5.84 (19, d, J=5.0Hz), 8.09 (1H, s) Preparation _3 3 To a solution of 7- [2,3-bis(tert- butoxycarbonyl) guanidino) -2 ,3-dihydro-lH-imidazo [1,2- 55 WO 2004/039814 PCT/JP2003/013684 b]pyrazole (1.83 g) in pyridine (10 ml) was added triphenylmethyl chloride (1.67 g) . The mixture was stirred at 50°C for 5 hours. After cooling, chloroform (50 ml) was added to the reaction mixture, and the 5 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 purified by 10 column chromatography on silica gel eluting with 2% methano1/chloroform to give 7- [2 ,3-bis (tert- butoxycarbonyl)guanidino]-l-triphenylmethyl-2,3-dihydr- lH-imidazo[1,2-blpyrazole (1.57 g) as a solid. 1H-NMR(CDC13) 1.47 (9H, s) , 1.48 (9H, s), 3.50 (2H, t, 15 J=7.8Hz), 3.92 (2H, t, J=7.8Hz), 7.07-7.26 (l0H, m), 7.53-7.54 (6H, m) , 8.34 (1H, brs), 11.12 (1H, brs) Example 16 To a solution of benzhydryl 7-[(Z)—2-(5-amino- 1,2,4-thiadiazol-3-yl) —2-(1-tert-butoxycarbonyl-l- 20 methylethoxyimin) acetamido]-3—iodomethyl—3-cephem—4— arboxylate (819 mg) in N,N-dimethylformamide (2.4 ml) as added N-(trimethylsilyl)acetamide (656 mg), and the ixture was stirred at room temperature for 30 minutes. To the reaction mixture was added 7-[2,3-bis(tert- 25 butoxycarbonyl)guanidino]-l-triphenylmethyl-2,3-dihydro- lH-imidazo[1,2-b]pyrazole (730 mg) . The whole mixture was stirred at room temperature for 6 hour a. To the resulting reaction mixture were added ethyl acetate (100 ml) and water (50 ml) . The aqueous layer was separated, 30 and the organic layer was washed with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into 35 diisopropyl ether (12 0 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene 56 WO 2004/039814 PCT/JP2003/013684 chloride (2.0 ml) were added anisole (O.67 ml) and trifluoroacetic acid (1.34 ml), and the mixture was stirred at room temperature for 4 hours. The reaction mixture was poured into diisopropyl ether (l20 ml). The 5 resulting precipitate was collected by filtration and dried in vacuo to give a crude product (430 mg) , 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 10 to about pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propsnol, The eXuate was concentrated to about 30 ml in vacuo and lyophilized to give 7-[(Z)-2-(5-amino-l,2,4-thiadiazol- 15 3-yl) -2-(l-carboxy-l-methylethoxyimino) acetamido]-3-[7- guanidino-2 ,3-dihydro-5- (lH-imidazo [1,2- b]pyrazolio)]methyl-3-cephem-4-carboxylate (20.4 mg) as an amorphous solid. IH-NMR(D2O) 1.51 (3H, s), 1.52 (3H, s), 3.35 (1H, d, 20 J=17.9Hz), 3.61 (1H, d, J=17.9Hz), 4,19 (2H, t, J=3.7Hz), 4.37 (1H, q, J=8.7Hz), 4.47 (1H, q, J=8.7Hz), 5.00 (1H, d, J=15.1Hz), 5.04 (1H, d, j=l5.1Hz), 5.26 (1H, d, J=4,8Hz), 5.84 (1H, d, J=4.8Hz), 8.13 (lH, s) Preparation 34 25 To a suspension of 1,1 '-carbonyldiimidazole (1.94 g) in methylene chloride (20 ml) was added tert-butyl N- (3-aminopropyl)carbamate (2.30 g), and the mixture was stirred at room temperature for 1 hour. To the reaction mixture were added N-ethyldiisopropylamine (2.56 g) and 30 4,5-diamino-l-methylpyrazole sulfuric acid salt (2.10 g) , and the mixture was stirred at 30oC for 3 days. The reaction mixture was concentrated in vacuo, The residue was purified by column chromatocgraphy on silica gel eluting with 6% methanol/chloroform to give 5-amino-4- 35 (3- (3-[(tert-butoxycarbonyl)amino]propyl]ureido)—1- methylpyrazole (1.75 g) as a solid. 1H-NMR(DMSO-d6) 1.37 (9H, s) , 1.43-1.49 (2H, m) , 2,89- 2.33 (2H, m) , 2.98-3.01 (2H, m) , 3.50 (3H, s) , 4.79 (2H, 57 WO 2004/039814 PCT/JP2003/013684 br) , 5.85 (H, br) 6.77 (1H, br), 6.96 (1H, s) , 7.12 (lH, br) Examaple 17 To a solution of benzhydryl 7-[(Z)-2-(5-tert- 5 butoxycarbonylamino-1-,2, 4-thiadiazol-3-yl) -2-(l-tert- butoxycarbonyl-l-methylethoxyimino) acetamido] -3- chloromethyl-3-cephem-4—carboxylate (1.0 g) in N,N- dimethylformamid (2.0 ml) was added sodium iodide (199 mg) , and the mixture was stirred at room temperature for 10 30 minutes. To the reaction mixture was added 5-aimino— 4— (3 — [3—[(tert-butoxycarbonyl)amino]propy1)ureido) - l- methylpyrazole (415 mg) and the whole mixture was stirred at 32°C for 24 hours. To the resulting reaction mixture were added ethyl acetate (50 ml) and water (50 15 ml) . The aqueous layer was separated, and the organic layer was washed with 10% aqueous sodium trifluoroacetate solution and brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate 20 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 (3.6 ml) were added anisole (1.2 ml) and trifluoroacetic acid (2.4 ml). The resulting 25 solution was stirred at room temperature for 4 hours and poured into diisopropyl ether (100 ml) . The resulting precipitate was collected by filtration and dried in vacuo to give a crude product (939 mg) , which was purified by preparative HPLC utilizing ODS column. The 30 eluate containing a desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to about pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical Corporation) eluting with 30% aqueous 2-propartol. The 35 eluate was concentrated to about 30 ml in vacuo and lyophilized to give 7-[ (Z) -2- (5-amino-l, 2 , 4-thiadiazol- 3-yl)—2—(1-carboxy-l-methylethoxyimino)acetamido]-3—(3— amino-4- [3- (3-aminopropyl) ureido] — 2-methyl-1- 53 WO 2004/039814 PCT/JP2003/013684 pyrazolio )methyl-3-cephem-4-carboxylate (53 mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s), 1.85-1.88 (2H, m) , 3.03 (2B, t, J=8Hz) , 3.22(2H, t, J=l8Hz), 3.26 (2H, 5 t, J=7Hz), 3.49 (1H, d, J=l8Hz), 3.72 (3H, s), 4.96 (1H, d, J=15Hz), 5.16 (1H, d, J=15Hz), 5.25 (1H, d, J=5Hz), Preparation 35 To a suspension of 1,1'-carbonyldiimidazole (973 10 mg) in methylene chloride (10 ml) was added tert-butyl N-(2-aminoethyl) carbamate (1.11 g) under ice-cooling, and the mixture was stirred at room temprature for 2 hours. To the reaction mixture were added N- ethyldiisopropylamnie (1.28 g) and 3-amino-4 ,5 , 6 , 7- 15 tetrahydropyrazolo[1,5-alpyrimidine sulfuric acid salt (1.18 g), and the mixture was stirred at 50oC for 6 hours. The reaction mixture was washed with brine. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo, The residue was 20 purified by column chromatography on silica gel eluting with 5% methanol/chloroform to give 3-(3-(2-[(tert- butoxycarbonyl)amino]ethyl)ureido)-4,5,6,7- tetrahydropyrazolo [1,5-alpyrimidine (150 mg) as a solid. 1H-NMR(CDCl3) 1.43 (9H, s) , 2.11-2.16 (2H, m) , 3.22- 25 3.35 (6H, m), 4.09 (2H, t, J=7Hz), 4.63 (lH, br), 5.14 (2H, br), 5.69 (1H, br), 7.17 (1H, s) Example 18 7 b -[(Z)-2-(5-Amino-l,2,4 thiadiazol-3-yl)-2-(l- carboxy-l-methylethoxyimino)acetamido]-3-(3-[3-(2- 30 aminoethyl ureidol-4 , 5 ,6 ,7-tetrahydro-1-pyrazol[1,5- al pyrimidinio ] methyl-3-cephem-4-carboxylate The title compound was obtained from benzhydryl 7 b -[(Z)-2-(5-tert-butoxycarbonylamino-l,2,4-thiadiazol- 3-yl)-2-(1-tert-butoxycarbonyl-l-methylethoxyimino)- 35 acetamidol-3-chloromethyl-3,-cephem-4-carboxylate and 3- (3-(2- [(tert-butoxycarbonyl)amino]ethyl]ureido)-4,5,6,7- tetrahydropyrazolo [1,5-alpyrimidine in the same manner as in Example 17 as an amorphous solid, 59 WO 2004/039814 PCT/JP2003/013684 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s), 2.09-2.21 (2H, m), 3.13 (2H, t, J=6Hz), 3.24 (1H, d, J=l8Hz), 3.35-3.52 (5H, m), 4,12-4.15 (2H, m), 4.88 (1H, d, J=16Hz), 5.13 (lH, d, J=16Hz) , 5.25 (1H, d, J=5Hz) , 5 . 85 (1H, d, 5 J=5Hz) , 7.8 3 (lH, s) Preparation 36 To a suspension of 1 ,1' -carboxyldiimidazole (97 3 mg) in methylene chloride (10 ml) was added 0-[2-(tert- butoxycarbonylamino) ethyl ]hydroxylamine (l.ll g) under 10 ice-cooling, and the mixtuire was stirred at room temperature for 2 hours. To the reaction mixture were added N-ethyldiisopropylamine (1.28 g) and 4,5-diamino- 1—methylpyrazole sulfuric acid salt (1.05 g) , and the mixture was stirred under reflux for 4 hours . The 15 reaction mixture was washed with 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 10% methanol/chloroform to give 5-amino-4—(3—(2- 20 [(tert-butoxycarbonyl)amino]ethoxy]ureido]-1- methylpyrazole (255 mg) as a solid. 1H-NMR(DMSO-d6) 1.38 (9H, s), 3.19-3.20 (2H, m) , 3.51 (3H, s), 3.72 (2H, t, J=6Hz) , 4.86 (2H, br), 6.95 (1H, br), 7.06 (1H, s), 8.02 (1H, brs), 9.15 (1H, brs) 25 Example 19 7 b [(Z)-2- (5-Amino-1,2, 4-thiadiazo1-3-yl) -2- 1 - carboxy- 1 -methylethoxyimino) acetamido ] -3 - (3—amino- 4 - [ 3 - (2-aminoethoxy)ureido]-2-methyl-l-pyrazolio)methyl-3- cephem-4-carboxylate 30 The title compound was obtained from benzhydryl 7 b -[(Z)—2-(5-tert-butoxycarbonylamino-1,2,4—thiadiazol- 3-yl) -2-(l-tert-butoxycarbonyl-l-methylethoxyimino) - acetamido]-3-chloromethyl-3-cephem-4-carboxylate and 5- amino-4-(3-[2-[(tert-butoxycarbonyl)amino] ethoxy)- 35 ureido)-1-methylpyrazole in the same manner as in Example 17 as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s) , 1.53 (3H, s), 3.21 (1H, d, J=18Hz), 3.33 (2H, t, J=5Hz), 3.47 (lH, d, J=18Hz), 3.74 60 WO 2004/039814 PCT/JP2003/013684 (3H, s), 4.17 (2H, t, J=5Hz), 4.59 (1H, d, J=l5Hz), 5.17 (1H, d, J=l5Hz), 5.26 (1H, d, J=5Hz), 5.8 6 (IH, d, J=5Hz), 7.93 (1H, s) Preparation 37 5 To a suspension of 1,1'-carbonyldiimidazole (1.95 g) in methylene chloride (20 ml) was added tart-butyl N- (2-aminoethyl)carbamate (1.92 g) under ice-cooling, and the mixture was stirred at room temperature for 2 hours. To the reaction mixture were added N- 10 ethyldiisopropylamine (2.59 g) and 7-amino-2,3-dihydro- lH-imidazo(1,2-b]pyrazole sulfucic acid salt (2.2.2 g) , and the mixture was stirred at room temperature for 16 hours. To the reaction mixture were added trityl chloride (9.0 g) and triethylamine (3.0 g) . The mixture 15 was stirred at room temperature for 24 hours. The reaction mixture was washed -with 10% aqxteous citric acid solution, brine and saturated aqueous sodium hydrogen carbonate solution, The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated 20 in Vacuo. The residue was purified by column chromatography on silica gel eluting with 3% methanol/chloroform to give 7-(3-(2-[(tert- butoxycarbonyl) aminolethyl]ureidol-2,3-dihydro-l- tritylimidazo [ 1, 2-b] pyrazole (8O0 mg) as a solid, 25 1H-NMR(CDCl3) 1.43 (9H, s) , 3.19 (4H, br) , 3.69 (1H, brs), 3.78-3.85 (4H, m), 4.51 [1H, br), 5.07 (lH, br), 7.20 (1H, s), 7.26-7.34 (9H, m), 7.45-7.47 (6H, m) Example 20 To a solution of benzhydryl 7- [(Z) -2- (5-amino- 30 1,2,4-thiadiazol-3-yl)-2-(l-tert-butoxycarbonyl-l-***** methyletho3iyimitio) acetamido] "3-iodorttethyl-3-cephenti"4" earboxylate (820 mg) in W,N-dimethylformamide (2^4 ml) was added W— (trimetliyl^ilyl) acetamMe (656 mg) , and the miHture was stirred at room temperature for 30 minutes, 35 To the reaction fliixture was added 7-t3-{2-[{tert- butoxycarbonyl)amino]ethyljureido)'2,3-dihydro-l- tritylimidaao[1,2-b]pyrasole [700 mg) , and the whole mixture was stirred at room temperature tor 6 hours. To 61 WO 2004/039814 PCT/JP2003/013684 the resulting reaction mixture were added ethyl acetate (50 ml) and water (50 ml), The aqueous layer was separated, and the organic layer was washed with 10% aqueous sodium trifluoroacetate solution and brine, 5 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 a solution of the 10 resulting solid in methylene chloride (3,0 ml) were added anisole (1.0 ml) and trifluotoacetic acid (2,0 ml). The resulting solution was stirred at room temperature for 4 hours and poured into diisopropyl ether (120 ml). The resulting precipitate was collected by filtration 15 and dried in vacuo to give a crude product, (830 mg) , 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 with concentrated. 20 hydrochloric acid and chromatographed on Diaion® 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 7 b -[(z)-2- (5-amino-l ,2 , 4-thiadiazol-3-yl) -2- (1-carboxy-l- 25 methylethoxyimino) acetamido] —3 —(7—(3— 2— aminoethyl)ureido] —2,3-dihydro—5-(1H-imidazo[1,2- b]pyrazolio) methyl-3-cephem-4-carboxylate (28.5 mg) as an amorphous solid. 1H-NMR(D2O) 1.53 (3H, s), 1.54 (3H, g), 3.14 (2H, t, 30 J=6Hz), 3.29 (1H, d, J=18Hz), 3.49 [2H, t, J=6HZ), 3.57 (1H, d, J=18Hz) , 4.15 (2H, t, J=9Hz) , 4.31-4.45 (2H, m), 4.94 (1H, d, J-l5Hz), 5.02 (1H, d, J=l5Hz), 5.27 (1H, d, J=5Hz), 5.85 (1H, d, J=5Hz) , 7.95 (1H, s) Preparation38 35 To a suspension of 1,1'-carbonyldiimidazole (2.0 g) in dehydrated chloroform (30 ml) was added a solution of tert-butyl N-(2-hydroxyethyl)carbamate (1.92 g) in dehydrated chloroform (10 ml) under ice-cooling, and the 62 WO 2004/039814 PCT/JP2003/013684 mixture was stirred at room temperature for 1 hour. To the reaction mixture were added N-ethyldiisopropylamine ( 2.2 ml) and 4.5-diamino- 1-methyl -pyrazole sulfuric acid salt (2.58 g) , and the mixture was stirred at room 5 temperature for 17.5 hours. To the reaction mixture were added trityl chloride (3.42 g) and triethylamine (1.25 g) . The mixture was stirred at roara temperature for 2 hours. The reaction mixture was washed with 10% aqueous citric acid solution., brine and saturated 10 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 5% methanol/chloroform to give 4-([2- (tert- 15 butoxycarbonylamino) ethoxycarbonyl ]amino)-5- tritylamino)-l-methylpyrazole (1.91 g) as a solid, 1H-NMR(CDCl3) b 1.46 (9H, s), 2.89 (3H, s), 3.30-3.36 (2H, m) , 4.03-4.07 (2H, m) , 4.37 (1H, brs), 4.75 £1H, br) , 5.42 (1H, br), 7.17-7,30 (l6H, m) 20 Example 21 7 b -[ (Z)-2- (5-Amino-l,2,4-thiadiazol-3-yl)-2- (1- carboxy-l-methylethoxyimin) acetamido] -3- (3-amino-4- [ (2- aminoethoxycarbonyl)amino]-2-methyl-l-pyrazolio)methyl- 3-cephem-4-carboxylate 25 The title compound was obtained from benzhydryl 7 b -((Z)-2-(5-amino-l,2,4—thiadiazol-3-yl)-2-(l-tert- butoxycarbonyl-1-methylethoxyimino)acetamido]-3- iodomethy 1-3-cephem-4-carboxylate and 4-{[2-(tert- butoxycarbony1amino) ethoxycarbonyl] amino)-5- 30 (tritylamino)l-methylpyrazole in the same manner as in Example 20 as an amorphous solid. 1H-NMR(D2O) b 1.53 (3H, s), 1.54 (3H, s), 3.18 (1H, d, J=l8Hz), 3.30-3.38 (2H, m), 3.43 (lH, d, J=15Hz), 3.71 (3H, s), 4.37-4.40 (2S, m), 4.97 (1H, d, J=15Hz), 5.18 35 (1H, d, J=15HZ) , 5.24 (1H, d, J=5Hz), 5.83 (1H, d, J=5Hz), 7.95 (1H, s) Preparation 39 To a solution of 7-amino-2 , 3-dihydro—1H- 63 WO 2004/039814 PCT/JP2003/013684 imidazo[l,2-blpyrazole sulfuric acid salt (1.42 g) and N-ethyldiisopropylamine (2.73 g) in methylene chloride (50 ml) was added N- [2- (tert-butoxycarbonylamino) - acetoxy]succinimide (1.90 g). The mixture was stirred 5 at room temperature for 22 hours. The reaction mixture was washed with saturated aqueous sodium hydrogen carbonate solution, and the organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo, The oily residue was purified by column 10 chromatography on silica gel eluting with 5% methanol/chloroform to give 7-[2-{tert- butoxycarbonylamino)acetyl]amino-2, 3-dihydro-lH- imidazo[1,2-b]pyrazole (1.07 g) as a solid. 1H-NMR(CDCl3) 1.47 [9H, S), 3.89 (2H, d, J=5.5Hz), 3.97 15 (2H, t, J=2.7, 7.3Hz), 4.18 (2H, t, J=7.3Hz), 4.55 (1H, br) , 5.22 (1H, br), 7.16 (lH, s), 7.95 (1H, br) Example 22 To a solution of benzhydryl 7- [ (Z) -- (5-tert- butoxycarbonylamino-1, 2 , 4-thiadiazol-3-yl} -2- (1-tert- 20 butoxycarbony1-1-methylethoxyimino)acetamido]-3- chlormethyl-3-cephem-4-carboxylate (1.0 g) in N,N- dimethylformamide (2.0 ml) was added sodium iodide (181 mg), and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added 7-[2- 25 (tert-butoxycarbonylamino}acetyl]amimo-2,3-dihydro-lH- imidazo [1,2-b]pyrazole (421 mg) , The whole mixture was stirred at 30°C for 3 Hours. To tixe resulting reaction mixture were added ethyl acetate (100 ml) and water (50 ml) . The aqueous layer was separated, and the organic 30 layer was washed with 10% aqueous sodium trifluoroacetate solution and brine, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (150 ml), and the 35 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 anlsole (1.0 ml) and trifluoroacetic acid (2.0 ml) . The resulting 64 WO 2004/039814 PCT/JP2003/013684 solution was stirred at room temperature for 4 hours and poured into diisopropyl ether (150 ml) . The resulting precipitate was collected by filtration and dried in vacuo to give a crude product (630 mg) , which was 5 purified by preparative HPLC utilising ODS column. The eluate containing a desired product, was concentrated to about 30 ml in vacuo. The concentrate was adjusted to about pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical 10 Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo and lyophilized to give 7[ (Z) -2-(5-amino-l, 2,4-thiadiazol- 3-yl} -2- (1-carboxy-l-methylethoxyimino) acetamido]-3- [7- (2-aminoacetamido) -2 , 3-dihydro-5- ( 1H-imidazo [1,2- 15 b] pyrazolio) ]methyl-3-cephem-4-carboxylate (20. 3 mg) as an amorphous solid. lH-NMR(D2O) 1.51 (3H, s) , 1.52 (3H, s) , 3.26 (2H, t, J=18Hz), 3.54 (2H, d, J=18Hz), 3.97 (2H, s), 4.16 (2H, t, J=9Hz) , 4.35 (1H, q, J=9Hz) , 4.44 (1H, q, J=9Hz), 4.97 20 (2H, d, J=15HZ), 5.04 (2H, d, J=l5Hz), 5.25 (lH, d, J=4Hz), 5.84 (1H, d, J=4Hz) , 8.10 (lH, s) Preparation 40 To a suspension of 4,5-diamino-l-(2- hydroxyethyl)pyrazole sulfuric acid salt (1.20 g) and N- 25 [2-(tert-butoxycarbonylamino) acetoxy] succinimide [l.35 g) in methylene chloride (20 ml) was added N- ethyldiisopropylamine (2.1 ml) under ice-cooling, and the mixture was stirred at room temperature for 17 hours. The reaction mixture was washed with water (4 0 ml) , 30 saturated aqueous sodium hydrogen carbonate solution (40 ml) and brine (40 ml). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The oily residue was purified by column chromatography on silica gel eluting with 10% 35 methanol/chloroform to give 5-amino-4- [2- (tert- butoxycarbonylamino) acetyl ] amino-1- ( 2- hydroxyethyl)pyrazole (l.20 g) as a solid- 1H-NMR(CDCl3) 1.46 (9H, 5), 3.89-3.90 (4H, m), 4.00- 65 WO 2004/039814 PCT/JP2003/013684 4.04 (2H, m, 4.26 (2R, br) , 5.51 (1H, br) , 7.17 (1H, s) , 8.06 (1H, br) Example 23_ 7 b - [ (Z) -2-(5-Amino-l,2,4-thiadiazol-3-yl) -2- (1- 5 carboxy-l-methylethoxyimino) acetamido]-3- [3-amino-4- (2- aminoacatamido) -2- (2-hydroxyethyl) -l-pyrazolio]methyl-3- cephem-4-carboxylate The title compound was obtained from benzhydryl 7 b -(Z)-2-(5-tert-butoxycarbonylamino-l,2,4-thiadiazol- 10 3-yl)-2-(1-tert-butoxycarbonyl-l-methylethoxyimino)- acetamido]-3—chloromethy1—3-cephem-4-carboxylate and 5— amino—4-[2- (tert-butoxycarbonylamino) acetyl] amino-1- (2- hydroxyethyl) pyrazole in the same manner as in Example 22 as an amorphous solid. 15 1H-HMR(D2O) 1.52 (6H, s), 3.15 (2H, d, J=l8Hz), 3.48 (2H, d, J=18Hz) , 3,88 (1H, dt, J=16Hz) 4.02 (2H, s) , 4.42 (1H, dt, J=16.5Hz), 5.07 (2H, d, J=15HZ), 5.13 (2H, d, J=15Hz), 5.27 (lH, d, J=5Hz), 5.84 {1H, d, J=5Hz), 8.09 {1H, s) 20 Preparation 41 To a solution of 3-amino-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrimidine sulfuric acid salt (2.96 g) and N-ethyldiisopropylamine (2.59 g) in methylene chloride (70 ml) was added N-[2- (tert— 25 butoxycarbonylamino)acetoxy]succinimide (2.72 g). The mixture was stirred at room temperature for 14 hours. The reaction mixture was washed with saturated aqueous sodium hydrogen, carbonate solution. The organic layer was dried over anhydrous magnesium sulfate, filtered and 30 concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with 6% methanol/chloroform to give 3-[2-(tert- butoxycarbonylamino) acetyl] amino-4 ,5,6,7- tetrahydropyrazolo[1,5—alpyrimidine (2.4 g) as a solid. 35 1H-NMR(CDCl3) 1.46 (9H, s) , 2.08-2.12 (2H, m) , 3.29- 3.32 (2H, m), 3.90 (2H, br), 4,07 (2H, t, J=6.0Hz), 5.00 (1H, br), 5,38 (1H, br), 7.12 (lH, s), 8.11 (1H, br) Example 24 66 WO 2004/039814 PCT/JP2003/013684 7 b -[(Z) -2-(5-Amino-l,2,4-thiadiazol-3-yl)-2- (1- carboxy-l-methylethoxyimino) acetamido ] -3- [3- (2- aminoacetamido) -4,5,6, 7-tetrahydro-l-pyrazolo [1,5- alpyrimidinio] methy1-3-cephem-4-carboxylate 5 The title compound was obtained from benzhydryl 7 b - [ (Z) -2- (5-tert-butoxycarbonylamino-l, 2 ,4-thiadiazol- 3-yl) -2- (1-tert.-butoxycarbonyl-l-methylethoxyimino)- acetamido]-3-chloromethyl-3-cephem-4-carboxylate and 3- [2- (tert-butoxycartibonylamino) acetyl) amino-4 ,5, 6,7- 10 tetrahydropyrazolo [1, 5-a ] pyrimidine in the same manner as in Example 22 as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s) , 1.53 (3H, s), 2.05-2.25 (2H, m) , 3.21 (2H, d, J=18Hz) , 3.45 (2H, d, J=18Hz) , 3.30- 3.45 (2H, m) , 4.00 (2H, s), 4.10-4.25 (2H, m) , 4.92 (2H, 15 d, J=15Hz), 5.17 (2H, d, J=15Hz) , 5.24 (1H, d, J=5Hz) , 5.84 (1H, d, J=5HZ), 7.97 (1H, s) Preparation 42 To a solution of 3-amino-4 , 5 , 6 ,7- "tetrahydropyrazolo [1 , 5-a] pyrimidine sulfuric acid salt 30 (4.44 g) and N-ethyldiisopropylamine (3.88 g) in methylene chloride (100 ml) was added N-[3-(tert- butoxycarbonylamino)propionyloxy) succinimide (4.29 g) . The mixture was stirred at room temperature for 6 hours. The reaction mixture was washed with saturated aqueous 25 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 5% methanol/chloroform to give 3-[3-(tert- 30 butoxycarbonylamino)propiony1]amino-4,5,6,7- tetrahydropyrazolo(1,5-alpyrimidine (3.67 g) as an oil. 1H-NMR(CDCl3) 1.43 (9H, s) , 2.08-2.13 (2H, m) , 2.52 (2H, t, J=6.0Hz), 3.32 (2H, t, J=5.0Hz), 3.43-3.46 (2H, m) , 4.07 (2H, t, J=6.0Hz, 5.12 (1H, br), 5.23 (1H, br), 35 7.13 (1H, s) , 7.97 (1H, br) Example 25 7 b - [ (Z) -2- (5-Amino-l,2 , 4-thiadiazol-3-yl) -2- (1- carboxy-l-methylethoxyimino) acetamido] -3- [3- (3- 67 WO 2004/039814 PCT/JP2003/013684 aminopropionamido) -4,5,6, 7-tetrahydro-l-pyrazalo [1,5- a] pyrimidinio] methyl-3-cephem-4-carboxyylate The title compound was obtained from benzhydryl 7 b -[ (Z) -2- (5-tert-butoxycarbonylamino-l ,2,4-thiadiazol- 5 3-yl) -2- (1-tert-butoxycarbonyl-1-methylethoxyimino)- acetamido]-3-chloromethyl-3-cephem-4-carboxylate and 3- [3- (tert-butoxycarbonylamino) propionyl]amino-4,5,6,7- tetrahydropyrazolo[1,5-a]pyrimidine in the same manner as in Example 22 as an amorphous solid. 10 1H-NMR(D2O) 1.51(3H,s), 1.52 (3H, s),2.05-2.25 (2H, m) ,2.85 (2H,t, J=7Hz),3.20 (2H, d,J=l8Hz),3.44 (2H, d, J=18Hz), 3.30-3.45 (2H,m),3.31 (2H, t, J=7Hz), 4.05-4.20 (2H, m), 4.91 (2H,d,J=16Hz), 5.16(2H,d, J=16HZ) ,5.23 (1H,d, J=5Hz), 5.84 (1H, d, J=5Hz) , 7.92 15 (1H, s) Preparaton 43 To a solution of 5-amino-methylpyrazole (100 g) in water (700 ml) were added concentrated hydrochloric acid (86 ml) and sodium nitrite (63.9 g) in water (200 20 ml) at a temperature below 10°C The reaction mixture was stirred at 5°C for 30 minutes. The precipitated solid was collected by filtration and dried to give 5- amino-l-methyl-4-nitrosopyrazole (117 g). 1H-NMR(DMSO-d6) 3.52 and 3.59 (3H, s), 7.22 and.8.51 25 (1H, s) , 8.17 and 8.51 (1H, brs) Preparation 44 To a suspension of 5-amino—1-methyl—4- nitrosopyrazole.(117 g) were added sulfuric acid (91 g) and 10% palladium on carbon (58 g). The mixture was 30 hydrogenatad under balloon pressure for 10 hours. The reaction mixture was filtered, and the filtrate was concentrated in vacuo. To the concentrate was added isopropyl alcohol (2.3 L), and the mixture was stirred for 1 hour. The precipitated solid, was collected by 35 filtration and dried to give 4,5-diamino—1- methylpyrazole sulfuric acid salt (158 g). 1H-NMR(D2O) 3.74 (3H, s), 7.80 (1H,s) Preparation 45 68 WO 2004/039814 PCT/JP2003/013684 A solution of 4, 5-diamino-1-methylpyrazole sulfuric acid salt (158 g) in water (l.l L) was neutralized to pH 6.9 with 4N aqueous sodium hydroxide solution, and dioxane (474 ml) was added to this 5 solution. To the resulting mixture was added dropwise phenyl chloroformate (124 g) maintaining pH of the mixture at 6.9 with 4N aqueous sodium hydroxide solution at a temperature below 10°C. The reaction mixture was stirred for 1 hour . The precipitated solid was 10 collected by filtration and dried to give 5-amino-l- methyl-4-phenoxycarbonylaminopyrazole (155 g) . 1H-NMR(DMSO-d6) 3.52 (3H ,s) , 5.00 (2H, brs) , 7.10-7.50 (6H, m) , 8.93 (1H, brs) Preparation 45 15 To a suspension of 5-amino-l-methyl-4- phenoxycarbonylaminopyrazole (153.8 g) in tetrahydrofuran (1 L) were added triethylamine (67 g) and triphenylmethyl chloride (185 g) at room temperature. The mixture was stirred for 6.5 hours. To the reaction 20 mixture was added heptane (2.6 L) , and the mixture was stirred for 1 hour. The precipitated solid was collected by filtration and washed with heptane- diisopropyl ether (1:1) . The crude solid was suspended in water (3 L), and the suspension was stirred for 1 25 hour. The solid was collected by filtration and dried to give l-methyl-4-phenoxycarbonyl-5- triphenylmethylaminopyrazole (253.6 g) . 1H-NMR(DMSO-d6) 2.74 (3H, s) , 5.57 (1H, brs) , 7.00-7.50 (21H, m), 8.12 (1H, brs) 30 Preparation 47 To a suspension of l-methyl-4- phenoxycarbonylamino-5-triphenymethylaminopyrazole (253.6 g) in N,N-dimethylformamide (l.5 L) were added triethylamine (59.5 g) and tert-butyl N-(2- 35 amino thyl) carbamate (94.2 g) in N,N -dimethylformamide (254 ml) . The mixture was stirred for 5 hours and poured into water (10.6 L). The slurry was stirred for 1 hour. The precipitated solid was collected by 69 WO 2004/039814 PCT/JP2003/013684 filtration and dried to give a crude product. The crude product was suspended in N,N-dimethylformamide, and the suspension was heated under reflux for 20 minutes. The suspension was cooled to ambient temperature over 4 5 hours. The solid was collected by filtration, washed with acetonitrile and dried to give 4-[N-(2-tert- butoxycarbonylaminoethyl) carbaramoylamino] -l-methyl-5- triphenylmethylaminopyrazole (261.2 g) . 1H-NMR(DMSO-d6) 2.69 (3H, s), 2.30-3.05 (4H, m) , 5.69 10 (1H, brs), 5.51-6.01 (1H, m), 6.74-6.81 (1H, m) , 6.87 (1H, brs) , 7.00 (1H, s), 7.10-7.30 (15H, m) Preparation 4 8 To a solution of (Z)-2-[5 -amino-1,2,4-thiadiazol- 3-yl) -2- (l--tert-butoxycarbonyl-l- 15 methylethoxyimino) acetic acid (319 g) in N,N- dimethylacetamide (1.5 L) were added potassium carbonate (113 g) and methanesulfonyl chloride (126 ml) under ice- cooling. The mixture was stirred at 10°C for 2 hours. The reaction mixture was added to a mixture of ethyl 20 acetate and water. The organic layer was washed with water and brine to give an activated acid solutiont On the other hand, a suspension of 4-methoxybenzyl 7- amino-3-chloromethyl-3-cephem- 4-carboxy late hydrochloride (3 0 0 g) in a mixture of water (1 L) and 25 ethyl acetate (1 L) was adjusted to pH 6 with triethylamine under ice-cooling. To the resulting mixture was dropwise added the above obtained activated acid solution, at 10oC under stirring. Stirring was continued at 5-10°C for 1.5 hours keeping pH of the 30 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 L), and the resulting precipitate was collected by filtration and dried to give 4- 35 methoxybenzyl 7-[(Z)-2-[5-amino-l,1,4-thiadiazol-3-yl)- 2-(1-tert-butoxycarbonyl-1-methylethoxyimino)acetamidol- 3-chloromethyl-3-cephem-4-carboxylate (495.7 g) . 1H-NMR(DMSO-d6) 1.39 (9H, s), 1.44 (6H, s), 3.45-3.70 70 WO 2004/039814 PCT/JP2003/013684 (2H, m) , 3.76 (3H, s) , 4.46 and 4.54 (1H, ABq, J=16HZ), 5.10-5.28 (2H+lH, m) , 5.90 (1H, dd, J=4 .9, 8.5Hz), 6.94 (2H, d, J=8.7Hz), 7.36 (2H, d, J=8.7Hz), 8.18 (2H, brs) , 9.52 (1H, d, J=5.5Hz) 5 Example 26 To a solution of 4-methoxybenzyl 7-[(Z)-2- (5- amino-1, 2 ,4-thiadiazol-3 -yl) -2 - (l-tert-butoxycarbonyl-l- methylethoxyimino) acetamidol -3-chloromethyl-3-cephem-4- carboxylate (150 g) in N,N-dimethylformamide (4 00 ml) 10 was added 1 ,3-bis (trimethylsilyl) urea (225 g) and the mixture was stirred for 30 minutes. Potassium iodide (51.2 g) was added to this solution, and the mixture was stirred for 30 minutes, 4-[N- (2-tert-Butoxycarbonylaminoethyl) - 15 carbamoylamino] -1 -methyl-5-triphenylmethylaminopyrazole (147 g) was dissolved in N,N-dimethylformamide (650 ml) at 78oC and the solution was cooled to 45°C, The solution was added to the solutian of 7-[ (Z)-2-(5- amino-1, 2 ,4-thiadiazol-3-yl) -2- (l-tert-butoxycarbonyl-l- 20 methylethoxyimino) acetamido] -3-chloromethy 1-3-cephem-4- carboxylate obtained above. The reaction mixture was stirred at 35°C for 18. 5 hours and poured into a mixture of ethyl acetate (21), water (1.8 L) and 20% aqueous sodium chloride solution (150 ml) , The organic layer 25 was washed with a mixture of 10% aqueous sodium thiosulfate solution (375 ml) and 20% aqueous sodium chloride solution (375 ml). The organic layer was washed successively with 10% aqueous sodium trifluoroacetate solution three times (750 ml x 3) and 30 20% aqueous sodium chloride solution (750 ml) . The organic layer was concentrated in vacuo , and the precipitated 4- [N- (2-tert-butoxycartbonylaminoethyl) - carbamoylamino]-l-methyl-5-triphenylmethylaminopyrazole was filtered off. The filtrate was further concentrated 35 in vacuo to a volume of approximately 6 00 ml. This solution was added to diisopropyl ether and the suspension was stirred for 1 hour. The resulting solid was collected by filtration and dried. The solid was 71 WO 2004/039814 PCT/JP2003/013684 dissolved in dichloromethane (669 ml) . To the solution were added anisole (223 ml) and trifluoroacetic acid (669 ml) . The reaction mixture was stirred for 4 hours and poured into diisopropyl ether. Tne resulting solid 5 was collected by filtration and dried. This solid was suspended in water, and pH of the suspension was adjusted to 1 with aqueous ammonia solution at a temperature below 10°C. The resulting precipitate was filtered off. The filtrate was acidified to pH 1 with 10 concentrated hydrochloric acid at a temperature below 10°C, and the resulting precipitate was filtered off. The filtrate was chromatographed on Diaion® HP-20 (11 L) eluting with 20% aqueous Z-propanol. The eluate was concentrated to about 3.5 L in vacuo, and 2M sulfuric 15 acid (51 ml) was added. The mixture was lyophilized to give a crude product (72.2 g) . The crude product (3 g) was purified by preparative HPLC utilizing ODS column. The eluate containing a desired product wag concentrated in vacuo. 20 The concentrate was adjusted to about pH 1 with concentrated hydrochloric acid and chromatographed on. Diaion® HP-20 (400 ml) eluting with 20% aqueous 2- propanol. The eluate was concentrated to about 73 ml in vacuo, and 2M sulfuric acid (1.5 ml) was added. The 25 mixture was further evaporated to a volume of approximately 12.5 ml, and water (6 ml) was added. After addition of seed crystals (10 mg), which resulted in the precipitation of a white solid, the mixture was stirred at room temperature for 1 hour. The mixture was 30 further stirred at 5°C for 13 hours. 2-Propanol £20 ml) as added at 5°C over 20 minutes , and the slurry was tirred at room temperature for 4 hours. 2-Propanol (20 l) was added over 30 minutes, and the slurry was tirred at room temperature for 3 hours. The 35 recipitated crystals were collected by filtration and ried to give 70-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)- 2—(l-carboxy-l-methylethoxyimiiio)acetamido]—3-(3-amino- 4- [H- (2-aminoethyl) cacbamoylamino] -2-methyl-l- 72 WO 2004/039814 PCT/JP2003/013684 pyrazolio)methyl-3-cephem-4-carboxylic acid hydrogen sulfate (1.51 g) as crystals. 1H-NMR(D2O) 1.61 (6H, s), 3.10-3.55 (6H, m), 3.71 (3H, 3), 5.02 and 5.23 (2H, ABq, J=16.7Hz), 5.25 (1H, d, 5 J=4.9Kz), 5.87 (1H, d, J=4.9Hz), 7.91 (lH, s) Preparation 49 A suspension of 4 , 5-diamino-l-methylpyrazole sulfuric acid salt (20 g) in triethylamine (29.2 ml) was stirred at 0°C for 10 minutes. A mixture of acetic 10 anhydride (9.87 ml) and formic acid (7.96 ml) was stirred at 4O°C for 30 minutes, cooled, to 0°C, and added dropwise to the above solution at 0°C. The whole mixture was stirred at 0°C for 2 hours. To the mixture was added brine, and the whole mixture was extracted 15 with tetrahydrofuran. The organic layer was dried over magnesium sulfate and evaporated under reduced pressure to give crude N—(5-amino-l-methyl-lH—pyrazol-4- yl) formamide, whicii was used in the next step without further purification. 20 Preparation 50 The crude product of N-(5-amino-l-methyl-lH- pyrazol—4-yl) formamide (13.33 g) was dissolved iri N,N- dimethylformamide (130 ml) . To the solution were added. trityl chloride (29.2 g) , triethylamine (66.3 ml) and 4- 25 dimethylaminopyridine (465 mg) , and the mixture was stirred at 60oC for 5 hours. To the reaction mixture was added water, and the whole mixture was extracted with ethyl acetate. The organic layer was washed with water and brine, and dried over magnesium sulfate. The 30 solvent was evaporated under reduced pressure to give a white solid. The solid was triturated with ethyl acetate/diisopropyl ether (1:1) to give N— [1-methyl-5— (tritylamino)-lH-pyrazol-4-yl] formamide (19.18 g) . The NMR spectrum of this compound indicates the existence of 35 its rotamer. lH-NMR(DMSO-d6) 2.76 and 2.92 (3H, s), 5.56 and 5.84 (1H, s), 7.0-7-4 (16H, m), 7.66 [1H, d, J=1.7Hz), 8.3- 8.4 (1H, m) 73 WO 2004/039814 PCT/JP2003/01368 ESI-MS: m/z =405.2(M+Na) + Preparation 51 To a solution of N- [l-methyl-5- (tritylamino) -1H- pyrazol-4-yl] formamide (3.825 g) in N,N- 5 dimethyl formamide (66 ml) was added sodium hydride (264 mg, 60% oil suspension) under a nitrogen atmosphere at 0oC under stirring. The mixture was stirred at 0oC for 15 minutes. To the mixture were added tert-butyl N-(3- bromopropyl) carbamate (2.62 g) in N,N-dimethylformamide 10 (10 ml) and sodium iodide (1.65 g) , The mixture was warmed to room temperature and stirred for 2 hours. 10% Aqueous potassium hydrogen sulfate solution (5 ml) was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with water and brine, and 15 dried over magnesium sulfate. The solvent was evaporated under reduced pressure, and the residue was chromatographed on silica gel eluting with methylene chloride/ethyl acetate (4:1) to give tert-butyl 3-(N- formyl-N-[l-methyl-5-(tritylamino)-lH-pyrazol-4- 20 yl]amino)propylcarbamate (2.714 g) . The NMR spectrum of this compound indicates the existence of its rotamer. 1H-NMR(DMSO-d6) 1.37 and 1-39 (9H, s) , 2.6-2.9 (6H, m) , 2.89 (3H, s), 5.34 and 6.01 (1H, s), 6.6-6-3 (lH, m) , 7.0-7.4 (l5H, m), 7.5-7.6 (1H, m), 7.95 (1H, s) 25 ESI-MS: m/z-5 62.3(M+Na)+ Example 27 7 b -[(Z)-2-(5-Amino-l,2,4-thiadiazol-3-yl)-2-(1- carboxy-1 -methylethoxyimino) acetamido] -3- (3 -amino-4- [N- (3-aminopropyl) -N-formylamino] -2-methyl-1- 30 pyrazolio)methyl-3-cephem-4-carboxylate The title compound was obtained from benzhydryl 7 b -[ (Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(l-tert- butoxycarbonyl-l-methylethoxyimino) acetamido] -3- iodomethyl-3-cephem-4-carboxylate and tert-butyl 3-(N- 35 formyl-N-[l-methyl-5-(tritylamino)-lH-pyrazol-4- yl] amino)propylcarbamate in the same manner as in Example 1. The NMR spectrum of this compound indicates the existence of its rotamer. 74 WO 2004/039814 PCT/JP2003/013684 lH-NMR(D2O) 1.53 (6H, s), 1.7-2.1 (2H, m) , 2.9-3.9 (9H, m) ,4.97 and 5.20 (2H, ABq, J=15.2Hz), 5.26 (1H, d, J=4.8HZ), 5.64 (1H, d, J=4.8Hz), 8.0-3.3 (2H, m) Example 28 5 To a suspension of 7-[(Z)-2-(5-amino-l,2,4- thiadiazol-3-yl) -2- (l-carboxy-1-methylethoxyimino)- acetamido]-3—(3—amino—4- [N- (3-aminopropyl)-N- formylamino]—2-methyl—1-pyrazolio]methyl-3-cephem—4- carboxylate (140 mg) in methanol (2.6 ml) was added 10 concentrated hydrocloric acid (0.176 ml) at room temperature, and the mixture was stirred for 6.5 hours. To the reaction mixture was added sodium hydrogen carbonate (177 mg), and the mixture was purified by preparative HPLC (ODS column, acetonitrile/phosphate 15 buffer (pH 7)=5:95). The eluate containing a desired product was evaporated to remove acetonitrile, acidified with diluted hydrochloric acid and chromatographed on Diaion® Hp-20 eluting with 20% aqueous 2-propanol. The eluate was concentrated under reduced pressure and 20 lyophilized to give 7-[(Z)-2-[5-amino-l,2,4-thiadiazol- 3-y1)-2-(1-carboxy-l-methylethoxyimino) acetamido] -3-(3- amino-4—[ (3-aminopropyl) amino] -2- methyl—1— pyrazolio}methyl-3-cephem-4-carboxylate (39 mg), 1H-NMR(D2O) 1.52-1.54 (6H, m), l.95 (2H, tt, J=7.3Hz, 25 7.3Hz), 3.0-3.2 (4H, m) , 3.16 and 3.38 (2H, ABq, J=17,7Hz), 3.68 (3H, s) , 4.89 and 5.11 (2H, ABq, J=15.6Hz), 5.22 (1H, d, J=4.3Hz), 5.83 (1H, d, J=4.8Hz), 7.59 (1H, s) ESI-MS: m/z=636.3(M-H)- 30 Preparation 52 tert-Butyl 2-{N-formyl-N-[1-methyl-5- (tritylamino) -lH-pyrazol-4-yl] amino] ethylcarbamate The title compound was obtained from N-[1-methyl- 5-tritylamino)-lH-pyrazol-4-yl]formamide and tert-butyl 35 N- (2-bromoethyl) carbamate in the same manner as in Preparation 51. IR(KBr) 1709, 1670, 1170, 704 cm-1 1H-NMR(DMSO-d6) 1.35 and 1.3 6 (9H, s), 2.65 and 2.7 5 75 WO 2004/039814 PCT/JP2003/013684 (3H, s), 2.73-2.90 (4H, m), 5.45 and 6.02 (1H, s), 6.78 and 6.33 (1H, t-like), 7.05-7.30 (15H, m), 7.31 and 7.57 (1H, s) ESI-MS: m/z=426.3 (M+H+) , 548.3(M+Na+) 5 Example 29 7 b -[ (Z)-2-(5-Amino-l,2, 4-thiadiazol-3-yl) -2-(1- carboxy-l-methylethoxyimino) acetamido] - 3-{3-amino-4- [N- (2-aminoethyl) - N-forylamino] -2-methyl-1- pyrazolio} methy1-3 -cephem-4-carboxylate 10 The title compound was obtained from benzhydryl 7 b [ (Z)-1-(5- amino-l,2,4-thiadiazol-3-yl)-2- (l-tert- butoxycarbonyl-l-methylethoxyimino)acetamido]-3- chloromethyl -3-cephem-4-carboxylate and tert-butyl 2-{N- formyl-N-[l-methyl-5-(tritylamino)-lH-pyrazol-4- 15 yl]amino}ethylcarbamate in the same manner as in Example 1. IR(KBr) 1770, 1675, 1653, 1597 cm-1 1H-NMR(DMSO-d6) 1.53 (6H, s), 3.12-3.78 (4H, m) , 3.77 and 3,78 (3H, s) , 3.86-3.96 (2H, m) , 5.00 and 5.19 (2H, 20 ABq, J= 15.2Hz), 5.28 (1H, d, J=4.8Hz), 5.86 (1H, d, J=4.8Hz), 8.15 and 8.18 (1H, s), 8.19 and 8.33 (lH, s) ESI-MS: m/z=652.2(M+H+) Example 30 7 b -[ (Z) -2-(5-Amino-l, 2 , 4-thiadiazol-3-yl)-2- (1- 25 carboxy-l-methylethoxyimino ) acetamido] -3 - [3-amino-4- [ (2- aminoethyl ) amino ] -2-methyl -1- pyrazolio] methyl -3-cephem- 4-carboxy1ate The title compound was obtained from 7-[ (Z)-2-(5- amino-1,2 , 4-thiadiazol-3-yl) -2- (1-carbozy-l- 30 methylethoxyimino)acetamido]-3-(3-amino-4-[N-(2- aminoethyl) -N-formylamino)-2-methyl-l-pyrazolio)metliyl- 3-cephem-4-carboxylate in the same manner as in Example 28 . IR(KBr) 1770, 1651, 1593 cm-1 35 1H-NMR(DMSO-d6) 1.53 (3H, s) , 1.59 (3H, s), 3.13-3.26 (4H, m), 3.26 and 3.39 (2H, ABq, J=17.8Hz), 3.68 (3H, s), 4.87 and 5.11 (2H, ABq, J=15.7Hz), 5.25 (1H, d, J=4.8Hz), 5.84 (1H, d, J=4.8Hz), 7.63 (1H, s) 76 WO 2004/039814 PCT/JP2003/013684 ESI-MS: m/z=622,2(M-H-) Preparation 53 To a suspension of l-methyl-lH-pyrazole-4, 5- diamine sulfate (86 g) in tetrahydrofuran (1.3 L) was 5 added triethylamine (117 ml), and then (2S)-4-[ (tert- butoxycarbonyl) amino]-2-hydroxybutanoic acid (82.5 g) was added to the mixture. To the mixture were added 1- hydroxybenzotriazole (58.3 g) and N-(3- dimethylaminopropyl) -N' -e.thylcarbodiimide hydrochloride 10 (82.7 g) under ice-coaling, The reaction mixture was stirred at room temperature for 8 hours. To the reaction mixture were added ethyl acetate (l.3 L) , saturated aqueous sodium hydrogen carbonate solution and sodium chloride, and the mixture was stirred for 30 15 minutes. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (1.0 L) six times. The extra was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel column 20 chromatography eluting with ethyl acetate/tetrahydrofuran (l/l) to give tert-butyl [(3s)- 4- [ (5-amino-l-methyl-lH-pyrazol-4-yl) amino] -3-hydroxy-4- oxobutyl} carbamate (69. 5g). 1H-NMR(CDC13) 1.43 (9H, s), 1.6-1.9 (1H, m) , 1.9-2.2 25 (1H, m), 3.1-3.3 (1H, m) , 3.3-3.5 (lH, m) , 3.65 (3H, s) , 4.20 (1H, dd, J=3.6, 6.6Hz), 4.7-5.3 (4H, m), 7.24 (1H, s) , 8.58 (1H, s) [b D(c=l.05, CHC13]-27.06° Preparation 54 30 To a solution of tert-butyl [(3S)-4-[{5-amino-l- methyl-lH-pyrazol-4-yl)amino]-3-hydroxy-4- oxobutyl}carbamate (68.-51 g) in N,N-dimethylformamide (350 ml) was added chlorotriphenylmethane (67 g). To the mixture was dropwise added triethylamine (6 7 ml) . 35 The mixture vas stirred at room temperature for 12 hours. The reaction mixture was dissolved in dichloromethane (2 L) . The solution was washed successively with water and brine. The extract was dried over anhydrous magnesium 77 WO 2004/039814 PCT/JP2003/013684 sulfate, filtered and concentrated in vacuo. The residue was triturated with acetonitrile and dried in vacuo to give tert-butyl [(3S)-3-hydroxy-4-{[l-methyl-5- (tritylamino)-lH-pyrazol-4-yl]amino]-4- 5 oxobutyl)carbamate (64 g). 1H-NMR(CDCl3) 1.46 (9H, s), 1.3-1.6 (1H, m) , 1.8-2.1 (1H, m), 2.95 (3H, s), 2.9-3.2 (1H, m) , 3.3-3.6 (1H, m), 3.95 (1H, m), 4.53 (1H, d, J=4.5Hz), 4.74 (1H, s), 4.92 (1H, brs), 7.1-7.3 (15H, m), 7.39 (1H, s), 7.73 (1H, s) 10 ESI-MS: m/z=6 38 .2(M+H+Na+) [b]20D{c=l.025, CHC13)=-36.5° Example 31 To a solution of 4-methoxybenzyl 7-[ (Z)-2—(5- amino-1,2,4-thiadiazol-3-yl) -2- (l-tert-butoxycarbonyl-l- 15 methylethoxyimino) acetamido] -3-chloromethyl-3-cephem-4— carboxylate (130 g) in N ,N- dimethyl formamide (400 ml) was added 1,3-bis(trimethylsilyl)urea (195 g) , and the mixture was stirred at room temperature for 30 minutes. To the solution was added potassium iodide (44.4 g), and 20 the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added tert-butyl ((3S) -3-hydroxy-4-{[l-methyl-5-(tritylamino)-lH-pyrazol- 4-yl]amino)-4-oxobutyl}carbamate (106 g), and the whole mixture was stirred at 35°C for 22 hours. To the 25 reaction mixture was added ethyl acetate (1.7 L), and the mixture was washed successively with water (1.6 L), 10% aqueous sodium trifluoroacetate solution (650 ml x 3) and brine (650 ml), dried over magnesium sulfate and filtered. The filtrate was concentrated to about 1 L in 30 vacuo. The concentrate was poured into diisopropyl ether (3 L), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the solid in methylene chloride (660 ml) were added anisole (220 ml) and trifluoroacetic acid (660 ml) . 35 The resulting solution was stirred at room temperature for 4 hours and poured into diiaopropyl ether (7 L) . The resulting precipitate was collected by filtration and dried in vacuo to give a cruds product 78 WO 2004/039814 PCT/JP2003/013684 (156.2 g). The crude product was dissolved in water (3.5 L). The solution was adjusted to about pH 3 with concentrated hydrochloric acid and chromatographed cm Diaion® HP-20 (Mitsubishi Chemical Corporation) eluting 5 with 20% aqueous 2-propanol. The eluate was concentrated to about 1.5 L in vacuo, and 2M aqueous sulfuric acid solution (33.18 ml) was added. The mixture was lyophilized. The lyophilised product (40 g) was dissolved in phosphate buffer (pH 7} and purified by 10 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 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical 15 Corporation) eluting with 10% aqueous 2-propanol. The eluate was concentrated to about 1 L in vacuo , and 2M aqueous sulfuric acid solution was added (13.59 ml). The resulting solution was lyophilized to give 7[(Z)- 2- (5-amino-l, 2 ,4-thiadiazol-3-yl) -2- (1-carboxy-l- 20 methylethoxyimino) acetamido] -3- {3-amino-4- [ [(2S)-4- amino-2-hydroxybutanoyl) amino] -2-methyl-l- pyrazolio)methyl-3-cephem-4-carboxylic acid hydrogen sulfate (20.32 g) as an amorphous solid, 1H-NMR(D2O) 1.61 (6H, s), 1.9-2.4 (2H, m), 3.20 (1H, d, 25 J=17.6Hz), 3.0-3.3 (2H, m), 3.45 (1H, d, J=l7.6Hz, 3.74 (3H, s), 4.47 (1H, dd, J=4, 6.3Hz), 5.06 (1H, d, J=15.7Hz), 5.25 (1H, d, J=4.8Hz), 5.28 (1H, d, J=15.7Hz), 5.87 (1H, d, J=4.8Hz), 8.07 (1H, s) Preparation 55 30 To a suspension of l-methyl-N5-trityl-lH-pyrazole- 4,5-diamine (1.60 g) in ethanol (50 ml) were added triethylamime (0.627 ml) and diethyl squarate (0.858 ml), and the mixture was stirred at room temperature for 22 hours. To the reaction mixture were added ethyl acetate 35 (200 ml) and hexane (100 ml), and the solution 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 79 WO 2004/039814 PCT/JP2003/013684 vacuo. The crystalline residue was washed with diethyl ether and dried in vacuo to give 3—ethoxy-4-{[1-methyl- 5-(tritylamino)-lH-pyrazol-4-yl]amino}-3-cyclobutene- 1,2-dione (1.45 g) as a solid. 5 1H-NMR(CDCl3) 1.42 (3H, br) , 2.99 (3H,s) , 4.41 (1H, brs), 4.69 (2H, q, J=7.2Hz), 6.40 (1H, br), 7.13-7.35 (16H,m) Preparation 56 To a suspension of tert—butyl 2- 10 aminoethylcarbamate (288 mg) and 3-ethoxy-4-{[l-methyl- 5-{tritylamino)-1H-pyrazol—4-yl]amino)-3-cyclobutene- 1,2-dione (718 mg) in ethanol (20 ml) was added triethylamine (0.209 ml), and the mixture was stirred under reflux for 4 hours. To the reaction mixture were 15 added diethyl ether and hexane. The crystalline precipitate was collected by filtration and dried in vacuo to give tert-butyl 2-[{2-([l-methyl-5- tritylamino)-lH-pyrazol-4-yl]amino)-3,4-dioxocyclobut- 1-en-l-yl)amino]ethylcarbamate (830 mg) as a solid. 20 1H-NMR(CDCl3) 1.40 (9H,s) 3.07-3-28 (5H, m) , 3.38- 3.67 (2H, m), 4.53-4.84 (lH, br), 4.84 (1H, br), 7.15- -7.22 (6H, m) , 7.23 (lH, s) , 7.22-7.34 (9H, m) Example 32 To a solution of benzhydryl 7-[(Z) -2- (5-amino- 25 1,2,4—thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-l- methylethoxyimino) acetamido] -3-iodomethyl-3-cephem-4- carboxylate (9 01 mg) in N,N-dimethylformamide (l.8 ml) was added N— (trimethylsilyl) acetamide (720 mg) , and the mixture was stirred at room temperature for 1 hour. To 30 the reaction mixture was added a solution of tert—butyl 2-[(2-{(l-methyl-5-(tritylamino)-lH-pyrazol-4-yl]amino}- 3 , 4-dioxocyclobut-l-en-l—yl) amino] ethylcarbamate (682 mg) in N,N-dimethylformamide (6.3 ml) , and the whole mixture was stirred at 35-40°C for 7 hours. To the 35 resulting reaction mixture was added ethyl acetate, and the precipitate was filtered off. The filtrate was washed successively with water and brine, dried over anhydrous sodium sulfate and filtered. The filtrate was 80 WO 2004/039814 PCT/JP2003/013684 concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (80 ml) , and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in 5 methylene chloride (2.6 ml) were added anisole (0.88 ml) and trifluoroacetic acid (2.6 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether (80 ml) . The resulting precipitate was collected by filtration and dried in 10 vacuo to give a crude product (580 mg) , 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 with concentrated hydrochloric acid and 15 chromatographed on Diaion 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 3-{[3-amino-4-({2-[(2- aminoethyl) amino]-3,4-dioxo-l-cyclobuten-l-yl)amino)-2- 20 methyl-1-pyrazoliolmethyl}-7-[(Z) -2- (5-amino-l ,2 , 4- thiadiazol-3-yl) -2- (l-carboxy-l- methylethoxyimino) acetatamido] -3-cephem-4-carboxylate (22 mg) as an amorphous solid. 1H-NMR(D2O) 1.53 (3H,s) 1.54 (3H, s), 3.26-3.36 (1H, 25 ml, 3.27 (2H, t, J=5.7Hz) , 3.58-3.69 (1H,m), 3.74 (3H, s), 3.86-4.03 (2H, m), 4.93 (lH, d, J=14.5Hz), 5.10 (1H, d, J=14.5HZ), 5.29 (1H, d, J=4.3Hz), 5.83 (1H, d, J=4.3Hz), 7.99 (lH, s) Preparation 57 30 To a suspension of tert-butyl 3- aminopropylcarbamate (366 mg) and 3-ethoxy-4-{1-methy1- 5- (tritylamino) —1H—pyrazol-4-yl] amino)-3-cyclobutene— l,2-dione (670 mg) in ethanol (30 ml) was added triethylamine (0.195 ml), and the mixture was stirred 35 under reflux for 3 hours, To the reaction mixture were added diethyl ether (40 ml) and hexane (10 ml). The crystalline precipitate was collected by filtration and dried in vacuo to give tert-butyl (3-[(2-[(l-methyl-5- 81 WO 2004/039814 PCT/JP2003/013684 (tritylamino) -lH-pyrazol-4-yl] amino }-3 ,4-dioxo-l- cyclobuten- 1-yl) amino] propyl} carbamate (783 mg) as a solid. 1H-NMR(CDC13) 1.43 (9H, s), 1.67 (2H, quintet, J=5.5Hz), 5 3.15 (2H, q, J=5.5Hz), 3.17 (3H, s) , 3.60 (2H, q, J=5.5Hz), 4.82 (1H, brs), 4.86 (1H, t, J=5.5Hz), 5.44 (1H, br), 5.86 (1H, br), 7.13-7.33 (15H, m), 7.17 (1H, s) Example 33 10 To a solution of benzhydryl 7-[(Z)-2-(5-amino- 1,2, 4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1- methylethoxyimino) acetamido] -3-iodomethyl-3—cephem-4— carboxylate (819 mg) in N,N-dimethylformamide (1.6 ml) was added N-trimethylsilyl) acetamide (656 mg) , and the 15 mixture was stirred at room temperature for 40 minutes. To the reaction mixture was added a solution of tert- butyl (3-[(2-{[l-methyl-5-(tritylamino)-lH-pyrazol-4- yl]amino}-3,4-dioxo-l-cyclobuten-l- yl) amino]propyl)carbamate (637 mg) in N,N- 20 dimethylformamide (3.2 ml) , and the whole mixture was stirrd at 35-40°C for 3.5 hours. To the resulting reaction mixture was added ethyl acetate (60 ml) and the precipitate was filtered off. The filtrate was washed successively with water (50 ml x 2) and brine (50 ml), 25 dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to about 8 ml in vacuo. The concentrate was poured into diisopropyl ether (80 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the 30 resulting solid in methylene chloride (2.4 ml) were added anisole (0.80 ml) and trifluoroacetic acid (1.6 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether (80 ml). The resulting precipitate was collected 35 by filtration and dried in vacuo to give a crude product (565 mg) , which was purified by preparative HPLC utilizing ODS column eluting with a mixture of acetonitrile and phosphate buffer (pH 5.5). The eluate 82 WO 2004/039814 PCT/JP2003/013684 containing a desired product was concentrated to about 20 ml in vacuo. The concentrate was desalted by prepative HPLC utilizing ODS column ,and the fraction eluted with 8 % acetonitrile/0.01 m hydrochloric acid was 5 concentrated to about 10 ml in vacuo and lyophilized to give 3-([3- amino-4-[(2-[(3-aminopropyl ) amino]-3,4-dioxo 1-cyclobuten-l-yl} amino) -2-methyl-l-pyrazolio] methyl} - 7 b -[ (Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(l-carboxy- 1-methylethoxyimino) acetamido) -3-cephem-4-carboxylate 10 trihydrochloride (34 mg) as an amorphous solid. 1H-NMR(D2O) 1.62 (6H, s), 2.02 (2H, quinter, J=7.3Hz), 3.09 (2H, t, J=7.3HZ), 3.32 (1H, d, J=17.5Hz), 3.54-3.65 (lH, m), 3.67-3.78 (2H, m) , 3.75 (3H, s), 4.93-5.23 (2H, m), 5.30 (1H, d, J=4.5Hz), 5.36 (1H, d, J=4.5Hz), 7.99 l5 (1H, s) Preparation 58 To a solution of 1,1-(1, 2-dioxo-l, 2- ethanediyl)bis-lH-imidazole (761 mg) in N,N- dimethylformamide (8 ml) was added l-methyl-N5-trityl- 20 lH-pyrazole-4,5-diamine (709 mg) under ice-cooling, and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added a solution of tert-butyl 2-aminoethylcarbamate (1.28 g) in N,N- dimethylformamide (2 ml) , and the mixture was stirred at 25 room temperature for 27 hours. To the reaction mixture was added ethyl acetate (50 ml) . After the precipitate was filtered off, the filtrate was washed successively with water, 5% aqueous citric acid solution and brine. The organic layer was dried over anhydrous sodium 30 sulfate, filtered and concentrated in vacuo. The crystalline residue was washed with a mixed solvent of diethyl ether and ethyl acetate and dried in vacuo to give tert-butyl {2-[ (2-{[l-methyl-5-(tritylamino)-1H- pyrazol-4-yl]amino}-2-oxoacetyl]amino}ethyl)carbamate 35 (823 mg) as a solid. 1H-NMR(CDC13) 1.43 (9H, s) , 2.97 (3H, s) . 3.31 (2H q, J=5.5Hz), 3.43 (2H, q, J=5.5HZ), 4.53 (1H, s), 4.84 (1H, brs) , 7. 10-7. 30 (15H, m) , 7.47 (1H, s) , 7.67 (1H, brs) , 83 WO 2004/039814 PCT/JP2003/013684 8.20 (1H, brs) Example 34 To a solution of 4-methoxybenzyl 7-[(Z)-2—{5- amino-1, 2 , 4-thiadiazol-3-Yl) -2- (l-tert-butoxycarbonyl-l- 5 methylethoxyimino) acetamido] - 3-chloromethyl-3-cephem-4- carboxylate (618 mg) in N,N-dimethylformamide (l.5 ml) was added N- (trimethylsilyl) acetamide (656 mg) , and the mixture was stirred at room temperature for 40 minutes. To the solution waa added potassium iodide (232 mg), and 10 the mixture was stirred at room temperature for 35 minutes. TO the reaction mixture was added a solution of tert-butyl (2-[(2-{[l-methyl-5-(tritylamino)-1H- pyrazol-4—yl] amino]-2-oxoacetyl)amino]ethyl]carbamate (626 mg) in N,N—dimethylformamide (3 ml), and the whole 15 mixture was stirred at 35-40°C for 24 hours. To the resulting reaction mixture was added ethyl acetate (50 ml) , and the solution was washed successively with water (50 ml x 2), 10% aqueous sodium trifluoroacetate solution (50 ml x 2) and brine (50 ml), dried over 20 anhydrous sodium sulfate and filtered. The filtrate was concentrated to about 10 ml in vacuo. The concentrate was poured into diisopropyl ether (60 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the solid in methylene 25 chloride (2.9 ml) were added anisole (0.95 ml) and trifluoroacetic acid (2.9 ml). The resulting solution was stirred at room temperature for 4 hours and poured into diisopropyl ether (60 ml) . The resulting precipitate was collected by filtration and dried in 30 vacuo to give a crude product (770 mg), 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 with concentrated hydrochloric acid and 35 chromatographad on Diaion® 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 3-{[3-amino-4-({2-[(2- 84 WO 2004/039814 PCT/JP2003/013684 aminoethyl) amino] -2-oxoacetyl} aininio) -2-methyl-l- pyrazolio]methyl] -7 b - [ (Z) -2-(5-amino-l , 2 , 4-thiadiazol-3- yl)-2-(1-carboxy-l-methylethoxyimino)acetamido]-3- cephem-4-carboxylate (31 mg) as an amorphous solid. 5 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s), 3.20 (1H, d, J=l8.0Hz), 3.24 (2H, t, J=6.0Hz), 3.45 (1H, dr J=18.0Hz ) 3.66 (2H, t, J=6.0Hz), 3.75 (3H, s) , 5.02 (1H, d, J=15.5Hz), 5,21 (1H, d, J=l5.5Hz), 5.25 (1H, d, J=5.0Hz), 5.85 (1H, d, J=5.0Hz), 8.14 (1H, s) 10 Preparaaation 59 To a suspension of phenyl {l-methyl-5- tert-butyl 3-azetidinylcarbamate acetic acid salt (418 mg) in methylene chloride (8 ml) was added N- 15 ethyldiisopropylamine (0.62 ml) , and the mixture was stitred under reflux for 16 hours. To the reaction mixture was added methylene chloride, and the solution was washed successively with 10% aqueous citric acid solution, 10% aqueous sodium hydroxide solution and 20 brine. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with a mixed solvent of ethyl acetate and hexane to give tert-butyl [1-([[1-methyl-5- (tritylamino)-lH-pyrazol-4—yl]amino ) carbonyl)-3 — 25 azetidinyl]carbamate (735 mg) as a solid, 1H-NMR(CDCl3) 1.47 (9H, s) , 2.92 (3H, s) , 3.56 (2H, dd, J=7.5, 5.0Hz), 4.02 (2H, dd, J=7.5, 7,5HZ), 4.42 (lH, brs), 4.71 (lH,s), 4.74 (1H, s), 4.94 (1H, brs), 7.18- 7.21 (7H,m) , 7.25-7.32 (9H, m) 30 Example 35 To a solution of benzhydryl 7-[ (Z)-2-(5-amino- l,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-l- methylethoxyimino) acetamido ] -3-iodomethyl-3-cephem-4- carboxylate (819 mg) in N,N-dimethylformamide (2.4 ml) 35 was added N-(trimethylsilyl) acetamide (655 mg) , and the mixture was stirred at room temperature for 3 0 minutes. To the reaction mixture was added a solution of tert- butyl [1-({[1-methyl-5-(tritylamino)-lH-pyrazol-4- 85 WO 2004/039814 PCT/JP2003/013684 yl] amino} carbonyl]— 3-acetidinyl] carbamate (553 mg) in N,N-dimethylformamide (3 ml) , and the whole mixture was stirred at room temperature for 3 hours, and then stirred at 50°C for 1 hour. To the resulting reaction 5 mixture were added ethyl acetate (50 ml) and water 50 ml) . The aqueous layer was separated, and the organic layer was washed with water and brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The 10 concentrate was poured into diisopropyl ether (80 ml) , and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene chloride (2.1 ml) were added anisole (0.7 ml) and trifluoroacetic acid (2.1 ml), 15 The resulting solution was stirred at room temperature for 4.5 hours and poured into diisopropyl ether (80 ml). The resulting precipitate was collected by filtration and dried in vacuo to give a crude product (521 mg) , which was purified by preparative HPLC utilizing ODS 20 column eluting with a mixture of acetonitrile and phosphate buffer (pH 5.5). The eluate containing a desired product was concentrated to about 20 ml in vacuo The concentrate was desalted by preparative HPLC utilizing ODS column, and the fraction eluted with 7% 25 acetonitrile/0.01 M hydrochloric acid was concentrated to about 10 ml in vacuo and lyophilized to give 3—[(3— amino-4-{ [ (3-amino-l-azetidinyl) carbonyl] amino] -2- methyl-1-pyrazolio) methyl]-7-[ (Z) -2- (5-amino-l,2,4- thiadiazol-3-yl)-2-(1-carboxy-l- 30 methylethoxyimino) acetamido]—3-cephem-4-carboxylate trihydrochloride (22 mg) as an amorphous solid. 1H-NMR(D2O) 1.62 (3H, s), 1.63 (3H, s), 3.25 (lH, d, J=17.9Hz) , 3.50 (1H, d, J=17.3Hz), 3.72 (3H, s) , 4.14 (2H, dd, J=9.6, 4.4Hz), 4.25 (1H, tt, J=7.8, 4.6Hz), 35 4.46 (2H, dd, J=9.6, 7.8Hz), 5.08 (1H, d, J=15.6HZ), 5.24 (1H, d, J=15.6Hz), 5.27 (1H, d, J=4.6Hz), 5.88 (1H, d, J=4.6HZ), 7.91 (lH, s) Preparation 60 86 WO 2004/039814 PCT/JP2003/013684 To a suspension of phenyl [l-methyl-5- (tritylamino) -lH-pyrazol-4-yl] carbamate (2. 18 g) and tert-butyl 3-amino-l-azetidinecarboxylate (7 93 mg) in methylene chloride (30 ml) was added N- 5 ethyldiisopropylamine (1.07 ml), and the mixture was stirred under reflux for 40 hours. To the reaction mixture was added methylene chloride, and the solution was washed successively with 10% aqueous citric acid solution, 10% aqueous sodium hydroxide solution and 10 brine. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by column chromatogrsphy on silica gel eluting with 10% methanol/methylene chloride to give tert-butyl 3- [{ (l-methyl-5- (tritylamino) -1H- 15 pyrazol-4—yl]amino)carbonyl)amino]-1- acetidinecarbonylate (1.52 g) as a solid. 1H-NMR(CDCl3) 1.44 (9H, s) , 3.03 (3H, s), 3.59 (2H, dd, J=9.2, 5.0Hz), 4.17 (2H, dd, J=9.2, 7.8Hz), 4.39-4.43 (3H, m) , 4.64 (lH, brs), 7.19-7.21 (1H, m) , 7.27 (1H,s), 20 7.29-7.32 (9H, m) Example 36 To a solution of 4-methoxybenzyl 7-[(Z)-2-(5- amino-1,2,4-thiadiazol-3-yl)-2-(l-tert-butoxycarbonyl-l- methylethoxyimino)acetamido]—3-chloromethyl-3-cephem-4- 25 carboxylate (1.48 g) in H,N-dimethylformaipide (3.0 ml) was added N- (trimethylsilyl) acetamide (1.42 g) , and the mixture was stirred at room temparature for 30 minutes. TO the solution was added potassium iodide (504 mg) and the mixture was stirred at room temperature for 30 30 minutes. To the reaction mixture was added a solution of tert-butyl 3-[({1-methyl-5-(tritylamino)-1H-pyrazol- 4—yl] amino } carbonyl) amino) -1-azetidinecarboxylate (1.20 g) in N,N-dimethylformamide (2.2 ml), and the whole mixture was stirred at 50°C for 16 Hours, To the 35 resulting reaction mixture was added ethyl acetate (200 ml), and the solution was washed successively with water (50 ml), 10% aqueous sodium trifluoroacetate solution (50 ml x 2) and brine (50 ml) , dried over anhydro-us 87 WO 2004/039814 PCT/JP2003/013684 sodium sulfate and filtered. The filtrate was concentrated to about 10 ml in vacuo. The concentrate was poured into diisopropyl ether (160 ml) , and tne resulting precipitate was collected by filtration and 5 dried in vacuo. To a solution of the solid in methylene chloride (8.64 ml) were added anisole (2.88 ml) and frifluoroacetic acid (8.64 ml). The resulting solution was stirred at room temperature for 3 hours and poured into diisopropyl ether (160 ml). The resulting 10 precipitate was collected by filtration and dried in vacuo to give a crude product (2.22 g), which was purified by preparative HPLC utilizing ODS column eluting with, a mixture of acetonitrile and phosphate buffer (pH 5.5). The eluate containing a desired 15 product was concentrated to about 20 ml in vacuo. The concentrate was desalted by preparative HPLC utilizing ODS column, and the fraction eluted with 8% aqueous acetonitrile was concentrated to about 10 ml in vacuo and lyophilized to give 3-[(3-amino-4-{[(3- 20 azetidinylamino)carbonyl)amino}—2-methy1-1- pyrazolio)methyl]-7-2-(5-amino-l,2,4-thiadiazol-3- yl)-1— (1—carboxy—1-methylethoxyimino)acetamido]-3- cephem—4—carboxylate (220 mg) as an amorphous solid. 1H-NMR(D2O) 1.50 (3H, s), 1.51 (3H, s), 3.20 (1H, d, 25 J=17.6Hz), 3.47 (lH, d, J=17.6Hz), 3.70 (3H, s), 4,18 (2H, dd, J=11.2, 7.6Hz), 4.31 (2H, dd, J=11.2, 8.3Hz), 4.68 (1H, tt, J=8.3, 7.6HZ), 4.94 (1H, d, J=l5.6Hz), 5.15 (1H, d, J=15.6HZ), 5.23 (1H, d, J=4.8Hz), 5.83 (1H, d, J=4.8Hz), 7.87 (1H, s) 30 Preparation 61 To a suspension of phenyl [l-methyl-5- (tritylamino)-lH-pyrazol-4-yl]carbamate (786 mg) and tert-butyl 3-pyrrolidinylcarbamate (373 mg) in methylene chloride (6 ml) was added N-ethyldiisopropylamine (0.43 35 ml), and the mixture was stirred under reflux for 10 hours. The reaction mixture was washed successively with 10% aqueous citric acid solution, brine and saturated aqueous sodium hydrogen carbonate solution. 88 WO 2004/039814 PCT/JP2003/013684 The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give tert-butyl {l-({[l-methyl-5- (tritylamino) -lH-pyrazol-4- yl] amino)carbonyl) -3-pyrrolidinyl] carbamate (730 mg) as 5 a solid. 1H-NMR(CDCl3) 1.43 (9H, s), 1.82-1.88 (1H, m) , 2.12- 2.18 (1H, m), 2.39 (3H, s), 2.89-3.03 (1H, m), 3.20-3.30 (2H, m) , 3.33-3.43 (1H, m) , 4.22 (1H, br), 4.69 (1H, br), 4.88 (1H, brs), 4.96 (1H, brs), 7.18-7.27 (16H, m) 10 Example 37 To a solution of benzhydryl 7[(Z)-2-(5-amino- l,2,4-thiadiazol-3-yl) -2- (1-tert-butoxycarbonyl-l- methylethoxyimino)acetamido]-3-iodomethyl-3—cephem—4— carboxylate (819 mg) in N,N-dimethylformamide (2.4 ml) 15 was added N- (trimethylsilyl) acetamide (655 mg) , and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added a solution, of tert- butyl [1- ({[1-methyl-5- (tritylamino)-lH-pyrazol-4- yl] amino} carbonyl) -3—pyrrolidinyl] carbamate (567 mg) in 20 N,N-dimethylformamide (3.0 ml) . The whole mixture was stirred at room temperature for 3 hoors. To the resulting reaction mixture were added ethyl acetate (100 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed successively with 10% 25 aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 2.5 ml in vacuo. The concentrate was poured into diisopropyl ether (80 ml), and the resulting precipitate 30 was collected by filtration and dried in vacuo. To a solution of the resulting solid in methylene chloride (2.55 ml) were added anisole (0.85 ml) and trifluoroacetic acid (2.55 ml), and the mixture was stirred at room temperature for 3 hours. The reaction 35 mixture was poured into diisopropyl ether (80 ml), and the resulting precipitate was collected by filtration, and dried in vacuo to give a crude product (6 08 mg) , which was purified by preparative HPLC utilizing ODS 89 WO 2004/039814 PCT/JP2003/013684 column eluting with a mixture of acetonitrile and phosphate buffer (pH 5.5), The eluate containing a desired product was concentrated to about 20 ml in vacua. The concentrated was desalted by preparative HPLC 5 utilizing ODS column, and the fraction eluted with 7% acetonitrile/0.01 M hydrochloric acid was concentrated to about 10 ml in vacuo and lyophilized to give 3- [ (3- amino-4- { [( 3-amino-l-pyrrolidinyl) carbonyl ) amino} -2- methyl-l-pyrazolio)methyl]-7-[(Z)-2-(5-amino-l,2,4- 10 thiadiazol-3-yl) -2- {1 -carboxy-1 - methylethoxyimino)acetamido]-3-cephem-4-carboxylate trihydrochloride (31 mg) as an amorphous solid. 1H-NMR(D20) 1.61 (3H, s), 1.61 (3H, s), 2.13-2.27 (1H, m) , 2.39-2.54 (1H, m) , 3.25 (1H, d, J=l8.lHz) , 3.51 (1H, 15 d, J=18.1HZ), 3.55-3.68 (3H,m), 3.73 (3H, s), 3.80 (1H, dd, j=ll.5, 6.0Hz), 4.01-4.11 (lH, m), 5.20 (1H, d, J=16.0Hz), 5.24 (1H, d, J=16.0Hz), 5.29 (1H, d, J=4.8Hz), 5.89 (1H, d, J=4.8Hz), 7.91 (1H, s) Preparation 62 20 To a suspension of phenyl [ 1-methy 1-5- (tritylamino)-lH-pyrazol-4-yl]carbamate (711 mg) and tert-butyl 3-amino-l-pyrrolidinecarboxylate (372 mg) in methylene chloride (15 ml) was added N- ethyldiisopropylamine (0.51 ml) , and the mixture was 25 stirred under reflux for 17 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 30 in vacuo, The residue was purified by column hromatography on silica gel eluting with 10% ethanol/methylene chloride to give tert-butyl 3-[{(l- ethy 1-5- (tritylamino) -lH-pyrazol-4- 1]amino ) carbonyl) amino]—l-pyrrolidinecarboxylate (511 35 mg) as a solid. 1H-NMR(CDCl3) 1.46 (9H, s) , 1.66-1.74 (1H, m) , 2.04- 2.11 (1H, m) , 2.97 (3H, s), 3.05-3.11 (lH, m) , 3.30-3.43 (2H, m) , 3.53-3.58 (1H, m), 4.16-4.23 (2H, m), 4.45 {1H, 90 WO 2004/039814 PCT/JP2003/013684 brs), 4.74 (1H, br), 7.18-7.20 (6H, m), 7.28-7.30 (10H, m) Example 3 8 To a solution of benzhydryl 7- [(Z) -2- (5-amino- 5 1,2 ,4-thiadiazol-3—yl) —2 — (1-tert-butoxycarbonyl—1- methylethoxyimino) acetamido] — 3— iodomethyl-3—cephem-4- carboxylate (707 mg) in N,N-dimethylformamide (2.1 ml) was added N-(trimethylsilyl)acetamide (566 mg), and the mixture was stirred at room temperature for 30 minutes. 10 To the reaction mixture was added a solution of tert- butyl 3- [ { { [l-methyl-5- (tritylamino) -lH-pyrazol-4- yl]amino) carbonyl)amino] -1-pyrrolidinecarboxylate (490 mg) in N,N-dimethylformamide (2.0 ml) , The whole mixture was stirred at room temperature for 3 hours. To 15 the resulting reaction mixture were added ethyl acetate (100 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed successively with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried 20 over sodium sulfate and filtered. The filtrate was concentrated to about 3 ml in vacuo. The concentrate was poured into diisoprbpyl ether (80 ml), and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the resulting solid in 25 methylene chloride (1.83 ml) were added aniaole (0.61 ml) and trifluoroacetic acid (1.83 ml), and the mixture was stirred at room temperature for 5 hours. The reaction mixture was poured into diisopropyl ether (80 ml), and the resulting precipitate was collected by 30 filtration and dried in vacuo to give a crude product (440 mg), which was purified by preparative HPLC utilizing ODS column. The eluate containing desired products was concentrated to about 30 ml in vacuo. The concentrate was adjusted to about pH 3 witli concentrated 35 hydrochloric acid and chromatographed on Diaion® 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-[(3- 91 WO 2004/039814 PCT/JP2003/013684 amino-2-methyl-4- {[ (3-pyrrolidinylamino) carbonyl] ami no} - 1-pyrazolio) methyl] -7- [ (Z) -2- (5-amino-l,2, 4-thiadiazol- 3-yl] -2-(1-carboxy-l-methylethoxyimino) acetamido] -3- cephem-4-carboxylate (18 mg) as an amorphous solid. 5 1H-NMR(D2O) 1.54 (3H, s) , 1.55 (3H, s), 2.00-2.10 (lH, m) , 2.30-2.40 (1H, m) , 3.23 (0.5H, d, J=17.9Hz), 3.24 (0.5H, d, j=l7.9Hz), 3.27-3.34 (1H, m), 3.34-3.43 (1H, m) , 3.45-3.57 (3H, m) , 3.7Z (3H, s) , 4.36-4.46 (1H, m) , 4.95 [0.5H, d, J=l5.lHz), 4.96 (0.5H, d, J=15.6Hz), 5.17 10 (1H, d, J=15. 6Hz), 5.26 (1H, d, J=5. OHz) , 5.85 (1H, d, J=5.0Hz), 7.88 (1H, s) Preparation 6 3 To a suspension of tert-butyl {2- [ ( ( [ 1-methyl-5- (tritylamino)-lH-pyrazol-4-yl]amino}carbonyl)amino]- 15 ethyl]carbaramate (10.8 g) in methanol (50 ml) was added 4M hydrogen chloride solution in dioxane (50 ml) . The mixture was stirred at room temperature for 3 hours. The solvent was concentrated in vacuo, and the residue was triturated with ethyl acetate and dried in vacuo to 30 give N-(2-aminoethyl)-N'-(5-amino-l-methyl-1H-pyrazo1-4- yl)urea trihydrochloride (5.6 g) as a solid. 1H-NMR(DMSO-d6) 2.84-2.87 (2H, m) , 3.30 (2H, brs), 3.71 (3H, 3), 6.57 (lH, br), 7.91 (1H, s), 3.05 (4H,br), 8.55 (1H, br) 25 Preparation 64 To a solution of N-(2-aminoethyl)-N ' - (5—amino-1- methyl-1H-pyrazol-4-yl) urea trihydrochloride (3.1 g) and triethylamine (4.6 g) in chloroform (100 ml) was added di-tert-butyl [{ (trifluoromethyl] sulfonyl] imino)- 30 methylene) biscarbamate (5.9 g) . The mixture was stirred at room temperature for 90 minutes. The reaction mixture was washed successively with 10% aqueous citric acid solution, brine and saturated, aqueous sodium hydrogen carbonate solution. The organic layer was 35 dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with ethyl acetate to give di-tert-butyl[ (Z)-{ [2-{{{5- amino-l-methyl-lH-pyrazol-4-yl) amino] carbonyl} amino} - 92 WO 2004/039814 PCT/JP2003/013684 ethy1]amino}methylidene)bis carbamate (4.3 g) as a solid. 1H-NMR(DMSO-d6) 1.39 (9H, s) , 1.48 (9H, s) , 3.18 (2H, q, J=6.0HZ), 3.35 (2H, br), 3.49 (3H, s), 4.77 (1H, brs), 6.05 (1H, br) , 6.97 (1H, s), 7,.9 (1H, brs) , 8.36 (1H, t, 5 J=5.5Hz), 11.49 (1H, brs) Preparation _65 j To a solution of di-tert-butyl { (Z)-{ [2-({[(5- amino-1-methyl-1H—pyrsazol-4-yl) amino] carbonyl ]amino)- ethyl] amino}methylidene) bis carbamate (2.2 g) and 10 triethylamine (0.6 g) in chloroform (3 0 ml) was added trityl chloride (1.7 g) , and the mixture was stirred at room temperature for 14 hours. The reaction mixture was washed successively with 10% aqueous citric acid solution, brine and saturated aqueous sodium hydrogen 15 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 di-tert-butyl [ (Z) - [ {2- [ ({ [l-methyl-5- (tritylamino)-lH-pyrazol-4-yl] amino] carbonyL} amino]- 20 ethyl}amino}methylidene]biscarbamate (1.9 g) as a solid. 1H-NMR(DMSO-d6) 1.39 (9H, s), 1.47 (9H, s), 2.72 (3H, s), 3.09-3.10 (2H, m), 3.31-3.34 (2H, m), 5.69 (1H, s), 6.10 (1H, br), 6.77 (1H, brs) , 7.02 (1H, s) , 7. 14-7.16 (6H, m) , 7.22-7.27 (9H, m), 3.36 (lH, t, J=5.5Hz), 11.51. 35 (1H, brs) Example 39 T:o a solution of benzhydryl 7 [ (Z) -2- (5—amino— l,2,4-thiadiazol-3-yl) -2- [l-tert-butoxycarbonyl-l- methylethoxyimino) acetamido) -3 - iodomethy 1-3-cephem-4- 30 carboxylate (820 mg) in N,N-dimethylformamide (1.4 ml) was added N-(trimethylsilyl) acetamide (6 56 mg), and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture were added di-tert-butyl [(Z)- ((2- [ ( { [1-methyl-5- (tritylamino) -lH-pyrazol-4- 35 yl ] amino} carbony 1) amino ] ethyl} amino) methylidane ] - biscarbamate (820 mg) and N,N--dimethylformamide (2.0 ml). The whole mixture was stirred at room temperature for 3 hours . To the resulting reaction mixture were added 93 WO 2004/039814 PCT/JP2003/013684 ethyl acetate (100 ml) and water (50 ml). The aqueous layer was separated, and the organic layer was washed successively with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and 5 brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuci. The concentrate was poured into diisopropyl ether (120 ml) , and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the 10 resulting solid 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 4 hours. The reaction mixture was poured into diisopropyl ether (100 ml) , and the resulting precipitate was 15 collected by filtration and dried in vacuo to give a crude product (740 mg), 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 20 pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® 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-{[3-amino-4-({[2- 25 guanidinoethyl)amino] carbonyl) amino) -2-methyl—1 — pyrazolio [methyl]-7- [ (Z) -2- (5-amino-l, 2 , 4-thiadiazol-3- yl) -2—(1-carboxy—1—methylethoxyimino)acetamido]-3— cephem-4-carboxylate (70 mg) as an amorphous solid. 1H-NMR(D2O) 1.55 (3H, s), 1.56 (3H, s), 3.24 (1H, d, 30 J=17.6HZ), 3.28-3.40 (4H, m), 3.52 (lH, d, J=17.5HZ), 3.73 (3H, s), 4.97 (1H, d, J=15.4Hz), 5.16 (1H, d, J=15.4Hz), 5.27 (1H, d, J=4.8Hz) , 5.8 4 (1H, d, J=4.8Hz), 7.37 (1H, s) Preparation 66 35 To a suspension of phenyl [1—methyl-5- (tritylamino)-lH-pyrazol-4-yl]carbamate (9 50 mg) and tert-butyl (3S)-3-pyrrolidinylcarbamate (560 mg) in methylene chloride (20 ml) was added, N- 94 WO 2004/039814 PCT/JP2003/013684 ethyldiisopropylamine (1350 mg), and the mixture was stirred under reflux for 23 hours. The reaction mixture was washed successively with 10% aqueous citric acid solution, brine and saturated aqueous sodium hydrogen 5 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 4% methanol/chloroform to give tert-butyl [ (3S) -1- { { [1- 10 methyl-5- (tritylamina) -1H-pyrazol—4-yl] amino] carbonyl)-. 3—pyrrolidinyll carbamate (680 mg) as a solid. 1H-NMR(CDCl3) 1.48 (9H,s) , 1.82-1.86 (1H, m) , 2.12- 2.18 (1H, m) , 2.89 (3H, s), 2.89-3.03 (1H, m), 3.20-3.30 (2Hr in), 3.3B-3.43 (IE, m) r 4 .22 (lHr br) t 4.69 [1M, bri) , 15 4.88 (lH, brs), 4.96 (1H, brs), 7.18-7.27 (16H, m) Example 40 To a solution of benzhydryl 7 [ (Z) -2- (5-amino- 1,2,4-thiadiazol-3-yl)-2-(1-tert-butoxycarbonyl-1- methylethoxyimino) acetamido] -3-iodomethyl—3-cephem-4- 20 carboxylate (320 mg) in N,N-dimethylformamide (2.4 ml) was added N-(trimethylsilyl)acetamide (656 mg), and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added tert-butyl [(3S)-1- ({[l-methyl-5-(tritylamino)-lH-pyrazol-4- 25 yl] amino}carbonyl)-3—pyrrolidinyl}-carbamate (680 mg). The whole mixture was stirred at room temperature for 3 hours. To the resulting reaction mixture were added ethyl acetate (80 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed 30 successively with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (120 ml), 33 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 , 95 WO 2004/039814 PCT/JP2003/013684 and the mixture was stirred at room temperature for 4 hours. The reaction mixture was poured into diisopropyl ether (10 0 ml), and the resulting precipitate was collected by filtration and dried in vacuo to give a 5 crude product (690 mg) , 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 with concentrated hydrochloric acid and 10 chromatographed on Diaion® 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-{{3-amino—4-({[(3S)-3—amino-1- pyrrolidinyl]carbonyl]amino)-2—methyl-1— 15 pyr azolio] methyl}-7 - [(Z)-2-95-amino-l, 2 , 4-thiadiazol-3- yl) -2- (l-carboxy-l-methylethoxyimino) acetamido] -3- cephem-4-carboxylate (60 mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (6H, s), 2.13-2.27 (1H, m), 2.38-2.53 (1H, m), 3.20 (1H, d, J=17.4Hz), 3.46 (1H, d, J=17.4Hz), 20 3.54-3.67 (3H, m) , 3.73 (3H, s), 3.79 (1H, dd, J=11.5, 6.0Hz), 4.00-4.10 (1H, m), 4.97 (1H, d, J=15.4Hz), 5.16 (1H, d, J=15.4Hz), 5.25 (lH, d, J=4.8Hz), 5.83 (1H, d, J=4.8Hz), 7.85(1H, s) Preparation 67 25 To a suspension of phenyl [ 1-methyl-5- (tritylamino)-1H—pyrazol-4-yl]carbamate (950 mg) and tert-butyl (3R)-3-pyrrolidinylcarbamate (5 60 mg) in methylene chloride (20 ml) was added N— ethyldiisopropylamine (39 0 mg), add the mixture was 30 stirred under reflux for 23 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 35 in vacuo. The residue was purified by column chromatography on silica gel eluting with 4% methanol/chloroform to give tert-butyl [(3R)-1-{{[1- methyl-5- (tritylamino) -lH-pyrazol-4-yl] amino ) carbonyl ) - 96 WO 2004/039814 PCT/JP2003/013684 3-pyrrolidinyl]carbamate (700 mg) as a solid. 1H-HMR(CDC13) 1.48 (9H, s), 1.82-1.88 (1H, m) , 2.12- 2.18 (1H, m), 2.39 (3H, s), 2.89-3.03 (lH, m) , 3.20-3.30 (2H, m) , 3.38-3.43 (1H, m) , 4.22 (1H, br) , 4.69 (lH, br) , 5 4.38 (1H, brs), 4.96 (1H, brs), 7.18-7.27 (16H, m) Example 41 To a solution of benzhydryl 7-[ (Z)-2-(5-amino- 1,2, 4-thiadiazol-3-yl) -2- (1-tert-butoxycarbonyl-l- methylethoxyimino) acetamido] -3-iodomethyl-3-cephem— 4- 10 carboxylate (820 mg) in N,N-dimethylformamide (2.4 ml) was added N-(trimethylsilyl)acetamide (656 mg), and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added tert-butyl [(3R)—1— ({[1—methyl-5—(tritylamino)-lH-pyrazo1-4— 15 yl] amino}carbonyl}-3-pyrrolidinyl] carbamate (680 mg) . The whole mixture was stirred at room temperature for 3 hours. To the resulting reaction mixture were added ethyl acetate (80 ml) and water (50 ml) . The aqueous layer was separated, and the organic layer was washed 20 successively with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried over sodium sulfate and filtered. The filtrate was concentrated to about 5 ml in vacuo. The concentrate was poured into diisopropyl ether (120 ml), 25 and the is suiting precipitate was collacted 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) , and the mixture was stirred at room temperature for 4 30 hours. The reaction mixture was poured into diisopropyl ether (100 ml), and the resultlug precipitate was collected by filtration and dried in vacuo to give a crude product (7 60 mg) , which was purified by preparative HPLC utilizing ODS column. The eluate 35 contaiuing a desired product was concentrated to about 30 ml in vacuo. The concentrate was adjusted to about pH 3 with concentrated hydrochloride acid and chromatographed on Diaion.® HP-20 (Mitsubishi Chemical 97 WO 2004/039814 PCT/JP2003/013684 Corporation) eluting with 30% aqueous 2-propanol. The eluate was concentrated to about 3 0 ml in vacuo and lyophilized to give 3-{[3-amino-4-{[(3R)-3-amino-l- pyrrolidinyl] carbonyl}amino) -2-methyl-1- 5 pyrazolio]methyl}-7-[(Z)-2-[5-amino-l,2,4-thiadiazol-3- yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3- (cephem-4-carboxylate (68 mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (6H, s) , 2.13-2.27 (1H, m) , 2.38-2.53 (1H, m) , 3.20 (1H, d, J=17.6Hz), 3.47 (1H, d, J=17.6Hz), 10 3.56-3.66 (3H, m) , 3.73 (3H, s), 3.79 (1H, dd, J=ll.0, 6.0Hz) , 4.00-4.10 (1H, m), 4.96 (1H, d, J=15.1Hz), 5.15 (1H, d, J=15.1Hz), 5.26 (1H, d, J=4.8Hz), 5.83 (1H, d, J=4.8Hz), 7.84 (1H, s) Preparation 68 15 To a suspension of phenyl (5-amino-l-methyl-lH- pyrazol-4-yl) carbamate (1.86 g) and (3s) -1-benzyl-3 - pyrrolidinamine (2.0 g) in chloroform (50 ml) was added N-ethyldiisopropylamine (3.1 g) , and the mixture was stirred under reflux for 19 hours. The reaction mixture 20 was concentrated in vacuo to give crude (S)—5-amino—4— [3-(l-benzyl-3-pyrrolidinyl)ureidol-l-methyl-lH-pyrazole as a solid. A solution of the crude product in acetic acid was treated with palladium black (3 ml) under a hydrogen atomosphere at room temperature for 24 hours. 25 After the catalyst was filtered off, the filtrate was concentrated in vacuo, and the residue was dissolved in saturated aqueous sodium hydrogen carbonate solution (100 ml) . To the solution was added a solution of di- tert-butyl dicarbonate (5.0 g) in tetrahydrofuran (40 30 ml} , and the mixture was stirred at room temperature for 5 hours. The reaction mixture was extracted with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. The residue was triturated with diethyl ether to give 35 tert-butyl (3S)-3-({ [ (5-amino-l-methyl-lH-pyrazol-4- yl) amino] carbonyl]amino] -1-pyrrolidinecarboxylate (1.9 g) as a solid. 1H-NMR(DMSO-d6) 1.40 (9H, s), 1.70-1.76 (1H,m) , 1.95- 98 WO 2004/039814 PCT/JP2003/013684 2.02 (1H, m) , 3.01-3.05 (1H, m) , 3.24-3.34 (2H, m) , 3.38-3.45 (1H, m) 3.50 (3H, s), 4.06-4.11 (lH, m), 4.78 (2H, brs), 6.19 (1H, brs) , 6.97 (1H, s) , 7.09 (1H, brs) Preparation 69 5 To a solution of tert-butyl (3S) -3- {( [ (5-amino-1- methyl-lH-pyrazol-4-yl)amino] carbonyl)amino}-1- pyrrolidinecarboxylate (1.8 g) and N- ethyldiisopropylamine (720 mg) in chloroform (50 ml) was added trityl chloride (1.6 g) , and the mixture was 10 stirred at room temperature for 28 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 15 concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with 3% methanol/chloroform to give tert-butyl (3S)-3-[({1- methyl-5- (tritylamino)-l-pyrazol-4- yl]amino)carbonyl)amino]-1—pyrrolidinecarboxylate (1.7 20 g) as a solid. 1H-NMR(CDCl3) 1.45 (9H, s), 1.66-1.74 (1H, m), 2.04- 2.11 (1H, m), 2.97 (3H, s), 3.05-3.11 (1H, m), 3.30-3.43 (2H, m), 3.53-3-58 (1H, m) 4.16-4.23 (2H, m) , 4.45 (1H, brs) , 4.74 (lH, br), 7.18-7.20 (6H, m) , 7.28-7.30 (10H, 25 m) Example 42 To a solution of benzhydryl 7- [ (Z) -2- (5-amino- 1,2, 4-thiadiazol-3-yl) -2- (1-tert-butoxycarbonyl-l- methylethoxyimino) acetamido ] -3-iodomethyl-3-cephem-4- 30 carboxylate (820 mg) in N,N-dimethylformamide (2.4 ml) was added N- (trimethylsilyl) acetamide (656 mg) , and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture was added tert-butyl (3S) -3- [ ({ [1-methyl-5- (tritylamino) -lH-pyrazol-4- 35 yl] amino} carbonyl) amino]-1-pyrrolidinecarboxylate (6 80 mg) . The whole mixture was stirred at room temperature for 3 hours. To the resulting reaction mixture were added ethyl acetate (80 ml) and water (50 ml) . The 99 WO 2004/039814 PCT/JP2003/013684 aqueous layer was separated, and the organic layer was washed successively with 10% aqueous sodium trifluoroacetate solution, 10% aqueous sodium thiosulfate solution and brine, dried over sodium 5 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 a solution of the resulting solid in 10 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 5 hours. The reaction mixture was poured into diisopropyl ether (10 0 ml), and the resulting precipitate was collected by filtration 15 and dried in vacuo to give a crude product (87 0mg) , 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 with concentrated 20 hydrochloric acid and chromatographed on Diaion® 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-{[3- amino-2—methyl—4—({[(3s)-3-pyirrolidinylamino]carbonyl}- 25 amino} -1-pyrazolio]methyl)-7- [(Z) -2- (5-amino-l ,2,4- thiadiazol—3-yl)-2-(1-carboxy-1-methylethoxyimino)- acetamido]-3-cephem-4-carboxylate (6 8 mg) as an amorphous solid. 1H-NMR(D2O) 1.52 (3H, s), 1.53 (3H, s), 2.00-2.09 (1H, 30 m) , 2.28-2.38 (1H, m) , 3.22 (1H, d, J=17.4Hz), 3.29 (1H, dd, J=12.4, 4.6Hz), 3.34-3.42 (1H, m), 3.44-3.54 (3H, m), 3.71 (3H, s), 4.36-4.43 (1H, m), 4.95 (lH, d, J=15,6Hz), 5.15 (1H, d, J=15.6Hz), 5.25 (1H, d, J=4.6Hz), 5.84 (lH, d, J=4.6Hz), 7.87 (1H, s) 35 Preparation_7 0 To a suspension of 4-[(tert— butoxycarbonyl) amino] butanoic acid (2.13 g) in dichloromethana (40 ml) was added l-hydroxybenzotriazole. 100 WO 2004/039814 PCT/JP2003/013684 (HOBT) (1.41 g) and N- (3-dimethylaminopropyl)-N' - ethylcarbodiimide hydrochloride (WSCD HC1) (3.65 g) , and the mixture was stirred for 1 hour. To the solution were added 1-methyl-lH-pyrazole-4,5-diamine sulfate (2 5 g) and N, N—diisopropylethylamine (3.32 ml), The reaction mixture was stirred for 18 hours. To the resulting solution were added brine and saturated aqueous sodium hydrogen carbonate solution, and the mixture was extracted with ethyl acetate. The aqueous 10 layer was extracted with tetrahydrofuran/ethyl acetate= 1/1 twice. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo. To the residue waS added pyridine (40 ml) , and then added chlorotriphenylmethane (5.3 g) , The mixture was 15 stirred at 65oC for 6 hours. The mixture was dissolved in ethyl acetate. Ths 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 20 residue was purified by column chromatography on silica gel eluting with 60% ethyl acetate/dichloronethane to give tert-butyl {4-{ [l-methyl-5- [tritylamiiao) -1H- pyrazol-4—yl] amino }-4—oxobutyl) carbamate (2.01 g) . 1H-NMR(CDCl3) 1.44 (9H, s) , 1.67 (2H, tt, J=6,7, 6.7Hz), 25 1.92 (2H, t, J=6.7Hz), 2.90 (3H, s), 3.09 (2H, dt, J=6.7, 6.7HZ), 4.50 (lH, s), 4.71 (1H, t, J=6.7Hz), 6.53 (1H, s) , 7.0-7.35 (16H, m) , 7.56 (1H, s) Example 43 To a solution of benzhydryl 7—[(Z) -2- (5-amino— 30 1,2 ,4-thiadiaEzl-3-yl)-2-(l-tert-butoxycarbonyl-l- methylethoxyimino) acetamido] -3-iodomethyl-3-cephem-4- carboxylate (2 g) in N,N-dimethylformamide (6 ml) was added N-(trimethylsilyl) acetamide (1.77 g) , and the mixture was stirred at room temperature for 30 minutes. 35 To the reaction mixture was added tert-butyl {4-{[1- methyl-5-{tritylamino)-lH-pyrazol-4-yl]amino}-4- oxobutyl)carbamate (1.93 g), and the whole mixture was stirred at 35oC for 30 hours. To the resulting reaction 101 WO 2004/039814 PCT/JP2003/013684 mixture was added ethyl acetate, and the solution was washed successively with water, 10% aqueous sodium trifluoroacetate solution and brine, dried over magnesium sulfate and filtered. The filtrate was 5 concentrated to about 25 ml in vacuo. The concentrate was poured into diisopropyl ether (150 ml) , and the resulting precipitate was collected by filtration and dried in vacuo. To a solution of the solid in methylene chloride (5 ml) were added anisole (1.5 ml) and 10 trifluoroacetic acid (5 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 (1.2 g). The crude product was dissolved 15 in a mixture of phosphate buffer (pH 6.86, 10 ml) and saturated aqueous sodium hydrogen carbonate solution and purified by preparative HPLC utilizing ODS column. The eluate containing a desired product was concentrated to about 20 ml in vacuo. The concentrate was adjusted to 20 about pH 3 with concentrated hydrochloric acid and chromatographed on Diaion® HP-20 (Mitsubishi Chemical Corporation) eluting with 20% aqueous 2-propanol. The eluate was concentrated to about 30 ml in vacuo, and 2M aqueous sulfuric acid solution (72 ml) was added. The 25 mixture was lyopliilized to give 3- { (3-amino-4-[(4- aminobutanoyl} amino] -2-methyl-l-pyrazolio}methyl) —7— [(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(1-carboxy-l- methylethoxyimino)acetamido]-3—cephem-4-carboxylic acid hydrogen sulfate (113 mg) as an amorphous solid. 30 1H-NMR(D2O) 1.61 (6H, s), 2.01 (2H, tt, J=7.6, 7.6Hz), 2.58 (2H, t, J=7.6Hz), 3.07 (2H, t, J=7.6Hz), 3.23 (1H, d, J=18Hz), 3.45 (1H, d, J=13Hz), 3.72 (3H, s), 5.06 (1H, d, J=15.7Hz), 5.25 (1H, d, J=4.8HZ), 5.28 (1H, d, J=15.7Hz), 5.87 (1H, d, J=4.8Hz), 8.03 (1H, s) 35 Preparation 71 tert-Butyl (5-{ [1-methyl-5-(tritylamino) -1H- pyrazol-4-yl] amino}-5-oxopentyl)carbamate The title compound was obtained from 5-[(tert- 102 WO 2004/039814 PCT/JP2003/013684 butoxycarbonyl) amino] pentanoic acid in the same manner as in Preparation 70. 1H-NMR(CDC13) 1.43 (9H, s) , 1.2-1.6 (4H, m) , 1.90 (2H, t, J=7.0Hz), 2.90 (3H, s), 3.09 (2H dt, J=7.0, 7.0Hz), 5 4.52 (lH, s) , 4.61 (1H, t, J=7.OHz), 6.28 (1H, s), 7.0- 7.35 (16H, m), 7.59 (1H, s) I Example 44 3 - ({3-Amino-4- [ {5-aminopentanoyl) amino] -2-methyl- l-pryazolio}methyl)-7-amino -1,2,4-thiadiazol- 10 3-y 1} - 2- (1 -carboxy-l-methylethoxyimino) acetamido] -3 - cephem-4-carboxylic acid hydrogen sulfate The title compound was obtained from tert-butyl (5- {[1-methyl-5- (tritylamino) -lH-pyrazol-4-yl ] amino) -5- oxopentyl) carbamate in the game manner as in Example 43 . 15 1H-NMR(D2O) 1.61 (6H, s) , 1.65-1.8 (4H, m) , 2.50 (2H, m), 3.03 (2H, m), 3.23 (1H, d, J=l6Hz), 3.45 (1H, d, J=l8Hz), 3.72 (3H, s), 5.06 (1H, d, J=15.7Hz), 5.25 (1H, d, J=4.8Hz) , 5.28 (lH, d, J=l5.7Hz), 5.87 (1H, d, J=4.8Hz), 8.02 (lH, s) 20 Preparation 72 To a solution of 1-methyl—N5-trityl-lH-pyrazole- 4,5-diamine (4 g) in dichloromethane (100 ml) was added tert-butyl 4- {[2, 5-dioxo-l-pyrrolidinyl) oxy] carbonyl ]- 1-piperidinecarboxylate (4.05 g), and the mixture was 25 refluxed for 72 hours. The reaction mixture 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 vaciao to give tert-butyl 4- { {1-methy 1-5- 30 (tritylamino)-lH-pyrazol-4-yl]amino]carbonyl)-1- piperidinecarboxylate (1.806 g). 1H-NMR(CDC13) 1.3-1.9 (14H, m) , l.5-1.8 (2H, m) , 2.95 (3H, s), 4.10 (2H, m) , 4.36 (1H,s), 6.53 (1H, s), 7.0-6 7.35 (16H, m) , 7.68 (lH, s) 35 Example_45 3- {(3-amino-2-methyl -4- {(4- piperidinylcarbonyl) amino] -l-pyrazolio}methyl) -7- [ (Z) - 2- (5-amino-l ,2 ,4-thiadiazol-3-yl) -2- (1-carboxy-l- 103 WO 2004/039814 PCT/JP2003/013684 methylethoxyimino) acetamido] -3-cephem-4-carboxylate The title compound was obtained from tet-butyl 4— ({[1-methy1-5-tritylamino)-lH-pyrazol-4- yl]amino}carbonyl) -l-piperidinecarboxylate in the same 5 manner as in Example 36. 1H-NMR(D2O) 1.57 (6H, s) , 1.8-2.3 (4H, m), 2.7-3.6 (7H, m) , 3.72 (3H, s) , 5.06 (1H, d, J=15.7Hz). 5,25 (1H, d, J=4.8Hz), 5.28 (1H, d, J=15.7Hz), 5.37 (1H, d, J=4.8Hz), 8.01 (1H, s) 10 Preparation 73 To a suspension of 3- [N- (tert-butoxycarbonyl )-K- methylamino]propanoic acid (3.33 g) in dicliloromethane (33 ml) and tetrahydrofuran (33 ml) were added HOBT (3.33 g) and WSC HCl (6.29 g) , and the mixture was 15 stirred for 1 hour. To the solution were added 1- methyl-lH-pyrazole-4 ,5-diamine sulfate (3.4 5 g) and N,N- diisopropylethylamine (11.4 ml). The reaction mixture was stirred at room temperature overnight. To the resulting solution was added brine and extracted with 20 tetrahydrofuran/ethyl acetate = 1/1. The extract was dried over anhydrous magnesium sulfate, filtered and concentrated in vacuo to give tert-butyl N-{3-[(5-amino- 1—methyl—lH—pyrazol-4—yl) amino] — 3—oxopropyl} —N- methylcarbamate as an oil (2.4 g) . This product was 25 used in the next step without further purification. Preparation 7 4 To a solution of tert-butyl N-{3-{5-amino-l- methyl-lH-pyrazol-4-yl)amine]-3-oxopropyl}-N- [methylcarbamate (4. 88 g) in N, N-dimethylformamide (50 30 ml} were added trityl chloride (6.86 g), triethylamine (6.86 ml) and 4-dimethylaminopyridine (80 mg) succesively. The mixture was stirred at room temperature overnight. To the resulting mixture was added ethyl acetate and washed with water (three times) 35 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 to give tert-butyl N-methyl-N-(3-{[1-methyl— 104 WO 2004/039814 PCT/JP2003/013684 5- (tritylamino) -lH-pyrazol-4-yl] amino} -3- oxopropyl)carbamate (4.20 g) as an amorphous solid. IR(KBr) 1659, 1587, 1491, 1446, 1173, 1151, 762, 739, 708 cm-1 5 1H-NMR(DMSO-d6) 1.40 (9H, s), 2.12 (2H, t, J=7.4Hz), 2.74 (3H,s) , 2.74 (3H, s), 3.24 (2H, t, J=7.4Hz), 5.58 (1H, s), 7.13-7.40 (16H, m), 8.30 (1H, s) Example 46 3- [ [3-Amino-2-methyl-4-{ [3- 10 (methylamino) propanoyl] amino ]-1-pyrazolio)methyl ] -7 [ (Z)-2- (5-amino-l,2,4-thiadiazol-3-yl) -2- (l-carboxy-1- methylethoxyimino) acetamido] -3 -cephem-4-carboxylate The title compound was obtained from tert-butyl N- methyl-N- (3-{ [l-methyl-5- (tritylamino) -lH-pyrazol-4- 15 yl] amino }— 3—oxopropyl) carbamate in the same msnner as in Example 32 as an amorphous solid, IR(KBr) 1770, 1664, 1599, 1531, 1400, 1360 cm-1 1H-NMR(D2O) 1.53 (6H, s) , 2.77 )(3H,s) , 2.92 (2H, t, J=6.5Hz), 3.13 and 3.45 (2H, ABq, J=17.7Hz), 3.74 (3H, 20 s) , 5.00 and 5. 21 (2H, ABq, J=15.4Hz), 5.25 (1H, d, J=4.8Hz), 5.35 (1H, d, J=4.8Hz), 8.02 (1H, s) ESI-MS 666.3 (M+H+) Preparation 7 5 tert-Butyl 3-{ [ {5-amino-l-methyl-lH-pyrazol-4- 25 yl) amino ) carbonyl) -1-azetidinecarboxylate The title compound was obtained from 1-(tert- butoxycarbonyl) -3-azetadinecarboxylic acid in the same manner as in Preparation 73 as an oil. This product was used in the next step without further purification. 30 Preparation 76 tert-Butyl 3-(([l-methyl-5-(tritylamino)-1H- pyrazol-4—yl) amino)carbonyl)-1—azetidinecarboxylate The title compound was obtained from tert-butyl 3- ( [(5-amino-l-methyl-lH-pyrazol-4-yl)amino] carbonyl}-1— 35 acetidinecarboxylate in the same manner as in Preparation 74 as an amorphous solid. IR(KBr) 3367, 3321, 1701, 1662, 1489, 1414, 1144, 766, 704 cm-1 105 WO 2004/039814 PCT/JP2003/013684 1H-NMR(DMSO-d6) 1.39 (9H, s), 2.75 (3H, s), 2.97-3.05 (1H, m) , 3.63-3.70 (2H, m) , 3.82-3.90 (2H, m) , 5.57 (1H, s), 7.10-7.33 (16H, m), 8.41 (1H, s) ESI-MS 560.3 (M+Na+) 5 Example 47 3- ( {3-Amino-4- [ {3-azetidinylcarbonyl) amino] -2- methyl-l-pyrazolio)methyl)-7 [(Z) -2- (5-amino-l, 2 ,4- thiadiazol-3-yl)-2-(1-carboxy-l- methylethoxyimino) acetamido} —3-cephem-4-carboxylate 10 The title compound was obtained from tert-butyl 3- ( {[l-methyl-5-(tritylamino)-lH-pyrazol-4- yl]amino)carbonyl)-l-azetidinecarboxylate in the same manner as in Example 32 as an amorphous solid. IR(KBr) 1768, 1663, 1624, 1605 , 1406, 1362 cm-1 15 1H-NMR(D2O) 1.53 (3H, s), 1.53 (3H, s) , 3.19 and 3.50 (2H, ABq, J=l7.7Hz), 3.82-3.98 (1H, m) , 4.31-4.35 (4H, m) , 4.49 and 5.20 (2H, ABq, J=15.3HZ), 5.25 (1H, d, J=4.8Hz), 5.85 (1H, d, J=4.8Hz), 8.04 (1H, s) ESI-MS 664.2 (M+H+) 20 Preparation 77 tert-Butyl N-(2-[ (5-amino-l-methyl-lH-pyrazol-4- yl) amino] -2-oxoethyl }-N-methylcarbamate The title compound was obtained from [H-(tert- butoxycarbonyl) -N- (methyl)amino] acetic acid in the same 25 manner as in Preparation 73 as an oil. This product was used in the next step without further purification. Preparation 7 6 tert-Butyl N-metyl-N-(2-{[-methyl-5- [tritylamino)-1H-pyrazol-4-yl]amino]-2- 30 oxoethyl)carbamate The title compound was obtained from tert-butyl N- {2-[(5-amino-l-methyl-lH-pyrazol-4-yl)amino]-2- oxoethyl}—N-methylcarbamate in the sams manner as in Preparation 74 as a white solid. The NMR spectrum of 35 this compound indicates the existence of its rotamer, 1H-NMR (DMSO-d6 ) 1.32 and 1.39 (9H,s) , 2.7 2 and 2.77 (3H, s) , 3.52 and 3.61 (2H, brs) , 5.61 (1H, s) , 7.13- 7.33 (16H, m), 8.20 and 8.30 (1H, brs) 106 WO 2004/039814 PCT/JP2003/013684 ESI-MS 548.3 (M+Na+) Example 48 3-[ {3-Amino-2-methyl-4- ([(methylamino)acetyl]amino)-1-pyrazolio)methyl]-7- 5 [ (Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2- (1-carboxy-l- methylethoxyimino} acetamido] -3-cephem-4-carboxylate The title compound was obtained from tert-butyl N- methyl-N- (2-{ [1-methy 1-5- (tritylamino) -lH-pyrazol-4- yl)amino}-2-oxoethyl) carbamate in the same manner as in 10 Example 32 as an amorphous solid. IR(KBr) 1770, 1657, l601, 1400, 1362 cm-1 lH-NMR(D2O) 1.53 (6H, s), 2.82 (3H, s), 3.18 and 3.45 (2H, Abq, J=17.7Hz), 3.74 (3H, s), 4.08 (2H, s), 5.00 and 5.20 (2H, ABq, J=15.3Hz), 5.25 (1H, d, J=4.8Hz), 15 5.84 (lH, d, J=4.8Hz), 8.05 (1H, s) ESI-MS 652.2 (M+H+) Proparation 79 N- (5-Amino-l-methyl-lH-pyrazol-4-yl) -2- (1 , 3-dioxo- 1,3-dihydro-2H-isoindol-2-yl) acetamide 20 The title compound was obtained from (1,3-dioxo- 1,3-dihydro-2H-isoindol-2-yl)acetic acid in the same manner as in Preparation 73 as a solid. 1H-NMR(DMSO-d6) 3.55 (3H, s), 4.36 (2H, S), 4.91 (2H, brs), 7.14 (1H, s), 7.85-8.02 (4H, m), 9.48 (1H, s) 25 ESI-MS 322.2 (M+Na+) Preparation 80 2-(1,3-Dioxo-l,3-dihydro-2H-isoindol-2-yl)-N-[1- methyl-5—(tritylamino)-lH-pyrazol-4—yl]acetamide The title compound was obtained from N-(5-amino-l- 30 methyl-lH-pyrazol-4-yl)-2-{1,3-dioxo-l,3-dihydro-2H- isoindol—2-yl)acetamide in the same manner as in Preparation 74 as aa solid. 1H-NMR(DMSO-d6) 2.70 (3H, s) , 4.12 (2H, 3) , 5.41 (1H, S), 7.12-7.33 (16H, m) , 7.85-7.95 (4H, m), 8.93 (1H, s) 35 ESI-MS 564.3 (M+Na+) Preparation 81 Hydrazine monohydrate (1.46 ml) was added to a solution of 2-{1,3-dioxo-l,3-dihydro-2H-isoindol-3-yl)- 107 WO 2004/039814 PCT/JP2003/013684 N-[l-methyl-5-(tritylamin)-lH-pyrazol-4-yl]acetamide (5.42 g) in ethanol (108 ml) and tetrahydrofuran (54 ml) at room temperature, and the mixture was stirred at 70°C for 2 hours. The reaction mixture was cooled to 0°C, 5 and the insoluble materials were removed by filtration. The filtrate was concentrated in vacuo. The residue was triturated with diisopropyl ether, collected by filtration and dried in vacuo to give 2-amino-N-[1- methyl-5-(tritylamino)-lH-pyrazo1-4-yl]acetatmide (3.3 7 10 g) as a solid. This product was used in the next step without further purification. Preparation 82 To a solution of 2—amino-N- [l-methyl-5- (tritylamino)-lH-pyrazol-4-yl]acetamide (2.47 g) in 15 tetrahydrofuran (50 ml) were added di-tert-butyl ({[(trifluoromethyl)sulfonyl]imino}metiiylene) - biscarbamate (2.35 g) and triethylamine (2.5 ml), and the mixture was stirred at room temperature for 30 minutes. The reaction mixture was poured into a mixture 20 of ethyl acetate and water. The aqueous layer was separated, and the organic layer was washed with brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated in vacuo. The concentrate was purified by silica gel column chromatography to give 25 di-tert-butyl { (E) -[(2-{[l-methyl-5-(tritylamino)-1H- pyrazol-4-yl] amino}-2-oxoethyl)amino]methylidene}- biscarbamate (3.25 g) as an amorphous solid. 1H-NMR(DMSO-d6) 1.38 (9H, s), 1.49 (9H, s), 2.75 (3H, s), 3.79 (2H, d, J=4.7Hz), 5.47 (1H, s), 7.12-7.33 (16H, 30 m), 8.55 (1H, t, J=4.7Hz), 8.61 (1H, s), 11.43 (1H, s) ESI-MS 676.3 (M+Na+) Example 49 3- ({3-Amino—4— [ (guanidinoacetyl) amino] —2—methyl—1- pyrazolio}methyl)-7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3- 35 yl) -2- (1-carboxy-l-methylethoxyimino) acetamido] -3- cephem- 4—carboxy1ate The title compound was obtained from di-tert-butyl { {E)-[ (2-{l-methyl-5-(tritylamino)-lH-pyrazol-4- 108 WO 2004/039814 PCT/JP2003/013684 yl] amino} -2-oxoethyl) ami no] methylidene )biscarbamate in the same manner as in Example 32 as an amorphous solid. IR(KBr) 1770, 1668, 1655, 1620, 1601, 1402, 1363 cm-1 1H-KMR(D2O) 1.53 (6H, s) , 3.20 and 3.48 (2H, ABq, 5 J=l7.6Hz, 3.75 (3H, s) , 4.21 (2H, s) , 5.00 and 5 . 20 (2H, ABq, J=15.3Hz) , 5.26 (1H, d, J=4 . 8Hz) , 5.85 (1H, d, J=4.8Hz) , 8.02 (1H, s) ESI-MS 678.2 (M-H+) (negative) Example 50 10 To a solution of 3-((3-amino-4-[(3- aminopropanoyl) amino]—2—methyl-1—pyrazolio }methyl- 7- [(Z) -2- [5-amino-l, 2 , 4-thiadiazol-3-yl) -2- (1-carboxy-l- methylethoxyimino)acetamido]-3-cephem-4-carboxylate (652 mg) in water (30 ml) and acetonitrile (3 ml) were added 15 ethyl formimidate hydrochloride (6 58 mg) and potassium carbonate (1.106 g) under ice cooling. After stirring at 5°C, for 3 hours , 1N HCl was added to neutralize the reaction mixture. The resulting solution was purified by preparative HPLC eluting with a mixture of phosphate 20 buffer (pH 5.5) and acetonitrile, and the eluate was subjected to column chromatography on Diaion© HP20 (Mitsubishi Chemical Corporation) and lyophilized to give 3- ( {3-amino-4- [ (3-guanidinopropanoyl) amino] -2- methyl-1-pyrazolio}methyl) -7[ (Z) -2- (5-amino-l , 2 ,4- 25 thiadiazol-3-yl)-2-(1-carboxy-l- methylethoxyimino) acetamidol]-3-cephem-4-carboxylate (23 mg) as an amorphous. The NMR spectrum of this compound indicates the existence of its rotamer. Only major isomsr was described. 30 IR(KBr) 1770, 1714, 1668, 1653, 1456, 1400 , 1360 cm-1 1H-NMR(D2O) 1.53 (6H, s), 2.85 (2H, t, J=6.4Hz), 3.19 and 3.46 (2H, ABq, J=l7.7Hz), 3.65 (2H , t, J=6.4Hz), 5.00 and 5.21 (2H, ABq, J=l5.2Hz), 5.26 (1H, d, J=4.8Hz), 5.85 (1H, d, J=4.8Hz), 7.80 (1H, s), 8.01 (1H, s) 35 ESl-MS 677.2 (M-H+) (negative) Preparation 83 To a stirred solution of l-methyl-lH-pyrazole-4,5- diamine sulfate (2.1 g) and 3-ethoxy-3-oxopropanoic acid. 109 WO 2004/039814 PCT/JP2003/013684 (1.32 g) in dichloromethane (10 ml) and tetrahydrofuran (10 ml) was added WSCD HCl (3.83 g) and N,N- diisopropylethylamine (6.96 ml), and the mixture was stirred overnight. The solvent was removed under 5 reduced pressure, and the crude residue which includes ethyl 3-[(5-amino-l-methyl-lH-pyrazol-4-yl)amino]-3- oxopropanoate was used for the next reaction without further purification. Preparation 84 10 The crude residue containing ethyl 3-[ (5-amino-l- methyl—1H—pyrazol-4-yl) amino] — 3-oxopropanoate was dissolved in N,N-dimethylformamide (20 ml), and trityl chloride (5.52 g) and triethylamine (4.14 ml) were added with stirring. The mixture was stirred overnight and 15 quenched with water (10 ml). The whole mixture was extracted with ethyl acetate, and the extract was washed with water and brine, dried over magnesium sulfate and concentrated, under reduced pressure to give a residual oil, which was chromatographed on silica gel eluting 20 with dichloromethane-ethyl acetate (2:3) to give ethyl 3-{[l-methyl-5-(tritylamino)-lH-pyrazol-4-yl]amino}-3- oxopropanoate (1,23 g) . ESI-MS 491.2 [M+Na]+ (positive), 467.3 (M-H]- (negative) 1H-NMR(DMSO-d6) 1.13 (3H, t, J=7.1Hz), 2.75 (3H, s), 25 3.04 (2H, s), 4.07 (2H, q, J=7.1Hz), 5.55 (1H, s), 7.1- 7.4 (16H, m), 8.54 (1H, s) Preparation 8 5 To a stirred solution of ethyl 3-[[l-methyl-5- (tritylamino) —lH-pyrazol-4-yll amino]-3-oxopropanoate 30 (1.3 g) in tetrahydrofuran (30 ml) was added 1N aqueous sodium hydroxide solution (3.1 ml), and the mixture was| stirred at room temperature for 3 hours. Tetrahydrofuran was removed in vacuo and the residue was made acidic with diluted citric acid. The resulting 35 precipitate was collected by filtration and dried under reduced pressure to give 3-{ (l-methyl-5-(tritylamino)- lH-pyrazol-4-yl] amino}-3-oxopropanoic acid (1.22 g) . ESI-MS 463.2 [M+Na]+ (positive) 110 WO 2004/039814 PCT/JP2003/013684 1H-NMR(DMSO-d6) 2.74 (3H s) , 2.95 ,(2H,s), 5.56 (1H, s) , 7.0-7.4 (16H, m), 8.54 (1H, s) , 12.0-13.0 (1H, brs) Preparation _86 To a suspension of 3-{ [l-methyl-5-(tritylamino)- 5 lH-pyrazol-4-yl) amino}-3-oxopropanoic acid (600 mg) and tert-butyl (2-aminoethyl)carbamate (240 mg) in tetrahydrofuran (12 ml) and dichloromethane (6 ml) was added WSCD HC1 (522 mg), and the whole mixture was stirred at room temperature overnight. To the reaction 10 mixturc was added water (3 ml) , and the whole mixture was extracted with ethyl acetate. The extract was washed with water and brine and dried over magnesium sulfate. The evaporation of the solvent gave a crude residue/ which was triturated with diisopropyl ether- 15 ethyl acetate (2:1) to give tert-butyl {2-[(3-{[1- methyl -5- (tritylamino ) -1H-pyrazol-4 -y 1 ] amino) -2- oxopropanoyl] amino] ethyl ) carbamate (53 7 mg) . ESI-MS 604.3 [M+Na]+ (positive) 1H-KMR(DMSO-d6) 1.38 (9H, s) , 2.74 (3H, s), 2 . 85 (2H, 20 s) , 2.9-3.2 (4H, m) , 5.61 (lH, s) , 6.7-6.3 (1H, m) , 7.0- 7.4 (16H, m), 8.0-8.1 (1H, m), 8.63 (lH, s) Example 51 3-{[3-Amino-4- [{3-[( 2-aminoethyl) amino}-3- oxopropanoyl] amino] - 2-methyl - 1-pyrazolio] methyl) –7b- 25 [ (Z) -2-(5-amino-l,2,4-thiadiazol-3-yl}-2-{l-carboxy-l- methylethoxyimino) acetamido] -3-cephem-4-carboxylate The title compound was obtained from tert-butyl {2-[{3-{[1-methyl-5-(tritylamino)-lH-pyrazo1-4- yl ]amino}-3-oxopropanoyl) amino] ethyl} carbamate in the 30 same manner as in Example 34. ESI-MS 731.2 [M+Na]+ (positive) 1H-NMR(D2O) 1.53 (6H, S) , 3.1-3.3 (2H, m), 3.19 and 3.44 (2H, ABq, J=l7.7Hz), 3. 54 (2H, s), 3.5-3.7 (2H, m) , 3.74 (3H, s) , 5.00 and 5.22 (2H, ABq, J=l5.5Hz), 5.25 35 (lH, d, J=4.7Hz), 5.86 (lH, d, J=4.8Hz), 8.05 (lH, s) Preparation 87 To a stirred solution of 3-amino-2- hydroxypropanoic acid (2.1 g) in tetrahydrofuran (30 ml) 111 WO 2004/039814 PCT/JP2003/013684 and water (30 ml) was added 1H aqueous sodium hydroxide solution to make the solution basic (pH=9) . To the mixture was added di-tert-butyl dicarbonate (4.36 g) , and the mixture was stirred at room temperature for 4 5 hours keeping pH of the mixture between 8.5 and 9.0. The whole mixture was washed with diethyl ether. The aquous layer was made acidic (pH=2) with 10% aqueous potassium hydrogen sulfate, saturated with sodium chloride and extracted with ethyl acetate. The extract 10 was dried over magnesium sulfate. The solvent was evaporated under reduced pressure to give 3-[(tert- butoxycarbonyl) amino]-2-hydroxypropanoic acid (3.96 g) . ESI-MS 228.2 [M+Na]+ (positive) 1H-NMR(DMSO-d6) 1.37 (9H, s), 3.0-3.8 (3H, m) , 3.9-4.1 15 (1H, m), 6.5-6.8 (lH, m) Preparation 88 To a solution of 3—[ (tert—butoxycarbonyl)amino] — 2 — hydroxypropanoic acid (1.61 gl in dichloromethane (8 ml) and tetrahydrofuran (8 ml) were added HOBT (1.59 g) and 20 WSCD HC1 (3.01 g) , and the mixture was stirred at room temperature for 1 hour. The solution was cooled to 0°C, and 1-methyl-lH-pyrazole—4 ,5-diamine sulfate and N,N— diisopropylethylamine (4.1 ml) were added. The mixture was stirred at room temperature for 8 hours. The 25 solvent was removed under reduced pressure to give crude tert-butyl {3—[(5—amino-1-methyl-1H-pyirazol-4-yl)amino]- 2-hydroxy-3-oxopropyl)carbamate, which was used in the next reaction without further purification. Preparation 89 30 tert-Butyl (2-hydroxy-3- { [l-methyl-5- (tritylamino)-lH-pyrazol-4-yl]amino}-3- oxopropyl)carbamate The title compound was obtained from tert-butyl [3- [ (5-amino-l-methyl-lH-pyrazol-4-yl) amino] -2-hydroxy- 35 3-oxopropyl}carbamate in the same manner as in Preparation 84. ESI-MS 564.3 [M+Na]+ (positive) 1H-NMR (DMSO-d6) 1.39 (9H, s) , 2.7-2.9 (1H, m), 2.83 (3H, 112 I WO 2004/039814 PCT/JP2003/013684 s), 3,1-3.4 (1H, m), 3.7-3.9 (1H, m), 5.79 (1H, d J=5.3Hz), 5.96 (lH, s), 6.5-6.7 (1H, m), 7.1-7.4 (16H, m) , 8.36 (1H, s) Example 52 5 3— { {3-Amino-4- [ (3-amino-l-hydroxypropanoyl} amino- 2-methyl-l-pyrazolio}methyl) - 7 - { (Z) -2- (5-amino-l ,2,4- thiadiazol-3-yl) -2- (1-carboxy-1- methylethoxyimino) acetamido] -3-cephem-4-carboxylate The title compound was obtained from tert-butyl 10 (2-hydroxy-3-{[l-methyl-5-(tritylaminol)-lH-pyrazol-4- yl] amino}-3-oxopropyl) carbamate in the same manner as in Example 32. 1H-NMR(D20) 1.49 (6H, s), 3.1-3.6 (4H, m) , 3.76 (3H, s), 4.6-4.7 (1H, m), 5.02 and 5.21 (2H, ABq, J=l5.4Hz), 5.26 15 (1H, d, J=4.8Hz), 5.86 (1H, d, J=4.8Hz) , 8.05 (1H, s) Preparation 90 To a suspension of 2- (4 , 5-dLamino-1H-pyrazol-l- yl) ethanol sulfate (5 g) in dichlorotmethane (50 ml] was added triethylamine (6.38 ml) at 0oC, and the mixture 20 was stirred at 0oC for 10 minutes. A mixture of acetic anhydride (2.16 ml) and formic acid (1.74 ml) was stirred at 40oC for 30 minutes, cooled to 0°C and added dropwise to the above solution at 0oC. The whole mixture was stirred at 0°C for 2 hours. To the mixture 25 was added brine, and the whole mixture was extracted with tetrahydrofuran. The extract was dried over magnesium sulfate and evaporated under reduced pressure to give crude [5-amino-l- (2-hydroxyethyl) -lH-pyrazol-4- yl]formamide, which was used in the next reaction 30 without further purification. Preparation_91 [1- (2-Hydroxyethyl) -5- (tritylamino) -lH-pyrazol-4- yl] formamide The title compound was obtained from [5-amino-l- 35 (2-hydroxyethyl)-lH-pyrazol-4-yl]formamide in the same manner as in preparation 84. The NMR spectrum of this compound indicates the existence of its rotamer. 1H-NMR(DMSO-d6) 3.10 (2H, t, J=6.2Hz), 3.3-3.5 and 3.4- 113 WO 2004/039814 PCT/JP2003/013684 3.6 (2H, m), 4.89 and 5.06 (1H, t, J=5.lHz), 5.77 and 6.07 (1H, s) , 7.1-7.4 (16H, m) , 7.58 and 8.07 (1H, s) , 7.58 (1H, s) Preparation 92 5 To a stirred solution of [1-(2-hydroxyethyl)-5— (tritylamino)-lH-pyrazol-4-yl]formamide (2 g) in N,N- dimethylformamide (30 ml) was added sodium hydride (213 mg, 60% oil suspension) under a nitrogen stream at 0°C, and the whole mixture was stirred at 0oC for 20 minutes. 10 A solution of tert-butyl (3-bromopropyl) carbamate (1.27 g) in N, N-dimethylformamide (10 ml) and sodium iodide (799 mg) were added to the above solution, and the mixture was stirred overnight. 10% Aqueous potassium hydrogen sulfate solution (5 ml) was added, and the 15 whole mixture was extracted with ethyl acetate. The extract was washed with water and brine and dried over magnesium sulfate. Evaporation of the solvent under reduced pressure gave an oil, which was chromatographed on silica gel eluting with dichloromethane-ethyl acetate 20 (2:1) to give tert-butyl (3-(N-formyl-N-[1-(2- hydroxyethyl)—5—(tritylamino)-1H—pyrazol—4— yl ] amino }propyl) carbamate (1 g) . The NMR spectrum of this compound indicates the existence of its rotamer. ESI-MS 592.3 [M+Na]+ (positive) 25 1H-NMR(DMSO-d6) 1.37 and 1.38 (9H, g) , 2.7-3.5 )(10H, m), 4.80 and 4.88 (lH, t, J=5.0Hz), 5.52 & 6.06 (1H, s), 6.5-6.9 (1H, m), 7.0-7.4 (16H, m), 7.52 (1H, s) Example 53 3— {{ 3—Amino-4- [N- (3-aminopropyl) -N-formylaminol -2- 30 {2-hydroxyethyl) -1-pyrazolio)methyl) -7- [ (Z) -2- (5-amino- l,2,4-thiadiazol-3-yl)-2-(1-carboxy-l- methylethoxyimirLO)acetamido]—3-csphem—4-carboxylate The title compound was obtained from tert-butyl (3-[N-formyl-H-[l-(2-hydroxyethyl)-5-(tritylamino)-1H- 35 pyrazol—4—yl]amino}propyl)carbamate in the same manner as in Example 32. ESI-MS 694.2 [M-H]- (negetive) 1H-NMR(D2O) 1.53 (6H, s) , 1.7-2.1 (2H, m) , 2.9-3.1 (2H, 114 WO 2004/039814 PCT/JP2003/013684 m) , 3.1-3.8 (4H, m) , 3.3-4.0 (2H, m) , 4.3-4 .6 (2H, m) , 4.3-5.2 (2H, m) , 5.29 (lH, d, J=4.8Hz), 5.85 (1H, d, J=4.7HZ), 8.0-8.3 (2H, m ) Example__54 . 5 To a stirred suspension of 3-({3-amino-4-[N-(3- aminopropyl) -N-formylamino]-2- (2-hydroxyethyl) -1- pyrazolio}methyl}-7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3- yl) -2- {1-carboxy-l-methylethoxyimino) acetamido] -3- cephem-4-carboxylate (100 mg) in methanol (l.4 ml) was 10 added concentrated hydrochloric acid (0.125 ml) at room temperature , and the mixture was stirred for 6.5 hours. To the above solution was added sodium hydrogen carbonate (109 mg), and the mixture was purified by preparative HPLC (ODS column; acetonitrile:phosphate 15 buffer (pH 7) = 5:95). The eluate containing a desired product was evaporated to remove acetonitrile, made acidic with diluted hydrochloric acid and chromatographed on Diaion® HP2 0 (Mitsubishi Chemical Corporation) eluting with 20% aqueous 2-propanol. The 20 eluate was concentrated under reduced pressure and lyophilized to give 3-{{3-amino-4-[(3- aminopropyl)amino)-2-(2-hydroxyethyl)-1- pyrazolio}methyl)-7-[ (Z) -2- (5-amino-l , 2 , 4-thiadiazol-3- yl)-2—(1-carboxy-l—methylethoxyimino)acetamido]-3- 25 cephem-4-carboxylate (18 mg) . ESI-MS 666.2 [M-H]- (negative) 1H-NMR(DMSO-d6) 1.53 (6H, s) , 1.96 (2H, tt, J=7.5Hz), 3.0-3,Z (4H, m), 3.13 and 3.43 (2H, ABq, J=17.6Hz), 3.87 (2H, t, J=4.8Hz), 4.2-4.4 (2H, m) , 4.87 and 5.03 (2H, 30 ABq, J=15.2Hz), 5.24 (1H, d, J=4.8Hz), 5.8 3 (lH, d, J=4.8 Hz) , 7.64 (1H, s) Preparation 93 To a stirred solution of N-[1-methyl-5- (tritylamino)-lH-pyrazol-4-yl]-2-(tritylamino)acetamide 35 (2 g) in N,N-dimethylformamide (20 ml) was added sodium hydride (245 mg, 60% oil suspension) at 0oC, and the mixture was stirred for 30 minutes with warming to room temperature. The mixture was cooled to 0oC, and methyl 115 WO 2004/039814 PCT/JP2003/013684 iodide (1.3 g) was added. The whole mixture was stirred at room temperature overnight. Water (5 ml) was added, and the whole mixture was extracted with ethyl acetate. The organic layer was washed with, water and birine and 5 dried over magnesium sulfate. Evaporation of the solvent under reduced pressure gave N-methyl—H-[1—methyl-5— (tritylamino)-lH-pyrazol-4-yl]-2-(tritylamino)acetamide (2.05 g). ESI-MS 690.3 [M+Na]+ (positive) 10 1H-NMR(DMSO-d6) 1.99 (3H, s), 2.3-2.S (3H, m) , 2.52 (3H, s), 5.44 (1H, s) , 6.85 (1H, s), 6.9-7.5 (30H, m) Preparation 94 Lithium aluminum hydride (4 55 mg) was added slowly to tetrahydrofuran (40 ml) at OoC and the mixture was 15 stirred for 20 minutes, N-Methyl-N-[l-methyl-5- (tritylamino)-lH-pyrazol-4-yl]-2-(tritylamino)acetamide (2 g) was added to the mixture at 0oC, and the whole mixture was stirred for 2 hours with warming to room temperature and refluxed for 2 hours. Sodium fluoride 20 (2.51 g) and water (8 62 mg) were added to the mixture, and the whole mixture was stirred at room temperature for 30 minutes. The precipitate was filtered off, and the filtrate was concentrated under reduced pressure to give crude residue, which was chromatographed (silica 25 gel; ethyl acetate:dichloromethane=l:10) to give N4,1- dimethyl-N5trityl-N4- [2- (tritylamino) ethyl] -1H- pyrazole-4,5-diamine (740 mg) . ESI-MS 676.2 [M+Na]+ (positive) 1H-NMR(DMSO-d6) 1.7-2.0 (2H, m) , 1.98 (3H, s), 2.2-2.4 30 (lH, m) , 2.6-2.8 (2H, m) , 2.81 (3H, s), , 5.24 (1H, s), 7.00 (1H, s), 7.0-7.5 (30H, m) Example 55 3- ( {3-Amino-4- [N- (2-aminoethyl) -N-methylamino] -2- methyl-l-pyrazolio}methyl)-7-[(Z)-2-(5-amino-l,2,4- 35 thiadiazol-3-yl)-2-(1-carboxy-l- methy1ethoxyimino)acetamido]-3-cephem-4-carboxy1ate The title compound was obtained from N4,l- dimethyl-N5-trityl-N4-[2-(tritylamino) ethyl] -lH- 116 WO 2004/039814 PCT/JP2003/013684 pyrazole-4, 5-diamine in the same manner as in Example 32. ESI-MS 636.2 [M-H]- (negative) 1H-NMR(D2O) 1.60 (6H, S) , 2.60 (3H, s), 3.0-3.2 (4H, m), 3.19 and 3.39 (2H, ABq, J=l7.7Hz), 3.67 (3H, s) 4.87 5 and 5.20 (2H, ABq, J=15.8Hz), 5.22 (1H, d, J=4.9Hz), 5.85 (1H, d, J=4.7Hz), 7.90 (1H, s) Preparation 95 To a solution of [1-(2-fluoroethyl)-lH-pyrazol-5- yl] formamide (15.7 g) in methanol (7 8 ml) was added 10 concentrated hydrochloric acid (21 ml) at room temperature. The reaction mixture was stirred for 3.5 hours and evaporated in vacuo . The residue was dissolved in ethyl acetate and washed with aqueous sodium hydrogen carbonate solution. The organic layer 15 was dried over magnesium sulfate and concentrated in vacuo to give 1- (2-fluoroethyl) -lH-pyrazol-5-amine (12 g). 1H-NMR(DMSO-d6) 4 . 15 (2H, dt, J=25 . 2 , 5.1HZ), 4.66 (2H, dt, J=47.2, 5.lHz), 5.1 (2H, brs) , 5.27 (lH, d, J=1.7HZ), 20 7.O6 (1H, d, J=1.7Hz) Preparation _9_6 To a solution of 1-(2-fluoroethyl)-lH-pyrazol-5- amine (12 g) in ethanol (30 ml) were added concentrated hydrochloric acid (70 mg) and isoamyl nitrite (10.9 g), 25 The reaction mixture was stirred at 25-38oC for 2 hours. Diisopropyl ether and hexane were added to the reaction mixture, and the resulting oil was purified by column chromatography on silica gel (ethyl acetate :hexane=l:2 à1:1 2:1 1:0) to give 1- (2-fluoroethyl) -4- 30 nitroso-lH-pyrazol-5-amine (4.8 g). 1H-NMR(DMSO-d6) 4.10-4.90 (4H, m), 7.09 and 8.59 (1H, s), 8.20 and 8.26 (1H, brs) Preparation 97 To a solution of 1-(2-fluoroethyl)-4—nitroso-lH— 35 pyrazol-5-amine (4,8 g) in water (30 ml) and methanol (30 ml) were added sulfuric acid (2.98 g) and 10% palladium on carbon (2.5 g), and the mixture was hydrogenatad under balloon pressure for 7.5 hours. The 117 WO 2004/039814 PCT/JP2003/013684 reaction mixture was filtered through a bed of Colite, and the filtrate was concentrated in vacuo. 2-Propanol was added to the residue, and the precipitate was collected by filtration to give 1-(2-fluoroethyl)-1H- 5 pyrazole-4 ,5-diamine sulfate (7 g). 1H-NMR(D2O) 4.25-4.95 (4H, m), 7.66 (1H, s) Preparation 9 8 To a suspension of 1-[2-fluoroethyl)-lH-pyrazole- 4,5-diamine sulfate (3 g) in tetrahydrofuran (30 ml) 10 were added tert-butyl {3-[(2,5-dioxo-l- pyrrolidinyl)oxy]-3—oxopropyl)carbamate (3.9 g) and ,N,N— diisoporpylethylamine (3.5 g) under ice-cooling. The reaction mixture was stirred at room temperature for 2 hours. An aqueous sodium hydrogen carbonate solution 15 and sodium chloride were added, and the mixture was extracted with ethyl acetate-tetrahydrofuran (three times). The organic layer was dried over magnesium sulfate and concentrated in vacuo. The residue was purified by column chromatography on silica gel (ethyl 20 acetate ethyl acetate:ethanol=8:1) to give tert-butyl (3-{ [5-amino-l- (2-fluoroethyl) -1H-pyrazol-4-yl] amino)-3- oxopropyl] carbamate (2.3 g) . 1H-NMR(DMSO-d6) 1.33 (9H, s), 2.36 (2H, t, J=7.1Hz), 3.10-3.27 (2H, m), 4.16 (2H, dt, J=25.5, 5.0Hz),4.67 (2H, 25 dt, =47.2, 5.OHz), 5.27 (2H, brs), 6.75-6.90 (1H, m) , 7.23 (1H, s), 9.03 (1H, brs) Preparation 99 To a solution of tert-butyl (3-{[5-amino-l-(2- fluoroethyl)-lH-pyrazol-4-yl]amino}-3— 30 oxopropyl) carbamate (2.3 g) in N,N-dimethylfomamide (12 ml) were added triethylamine (1.48 g), 4- dimethylaminopyridine (35.6 mg) and trityl chloride (2.2 g) at room temperature. The reaction mixture was stirred for 2 hours, and water was added. The mixture 35 was extracted with ethyl acetate, and the organic layer was washed with, water and aqueous sodium chloride solution. The organic layer was dried over magnesium sulfate and concentrated in vacuo. Acetonitrile was 118 WO 2004/039814 PCT/JP2003/013684 added, and the precipitate was collected by filtration to give tert-butyl {3-([l-(2-fluoroethyl)-5- ( tritylamino)-1H-pyrazol-4-yl] amino }-3- oxopropyl)carbamate (2 g) . 5 1H-NMR(DMSO-d6) 1.39 (9H, s), 2.05 (2H, t, J=7.2Hz), 3.00-3.03 (2H, m), 3.23 (2H, dt, J=25.3, 5.1Hz), 4.41 (2H, dt, J=47.1, 5.1Hz) Example 56 3- {( 3-Amino-4- [(3-aminopropionyl) amino] -2- {2- 10 fluroethyl)-l-pyrazolio)methyl)-7-[ (Z)-2-(5-amino- 1,2,4-thiadiazol-3-yl}-2-(1-carboxy-l- methylethoxyimino) acetamido ] -3-cephem-4 - carboxy1ate The title compound was obtained from tert-butyl (3- {(1-(2-fluroethyl)-5- (tritylamino) -lH-pyrazol-4- 15 yl] amino }-3-oxopropyl) carbamate in the same manner as in Example 38. 1H-NMR(D2O) 2.89 (2H, t, J=6.5Hz), 3.22 (1H, d, J=9.2Hz), 3.34 (1H, t, J=6.5Hz), 3.50 (1H, d, J=9.2Hz), 4.55-4.95 (4H, m), 5.08 (2H, brs), 5.26 (1H, d, J=4.9Hz), 20 5.84 (1H, d, J=4.9Hz), 8.09(1H,s) Preparation 100 l-Methyl-7-nitroso-lH-imidazo [1, 2-b]pyrazole The title compound was obtained from 1-methyl-lH- imidazo [1,2-b]pyrazole in the same manner as in 25 Preparation 96. 1H-NMR(DMSO-d6) 3.93 (1H, s), 7.43 (1H, m) , 7.92 (1H, m), 9.03 (1H, s) Preparation 101 1 -Methyl-lH-imidazo[l,2-b]pyrazol-7- amine sulfate 30 The title compound was obtained from l-methyl-7- nitroso-1H-imidazo[l,2-b]pyrazole in the same manner as in Preparation 97. 1H-NMR (DMSO-d6) 3.73 (3H, s) , 7.24 (1H, m), 7.62 (2H, m) 35 Preparation 102 Di-tert-butyl 1(Z)-[(1-methyl-lH-imidazo[1,2- b]pyrazol-7-yl) amino]methylidene}biscarbamate The title compound was obtained from 1—methyl-lH- 119 WO 2004/039814 PCT/JP2003/013684 imidazo [ 1,2-b] pyrazol-7-amine sulfate in the same manner as in Preparation 64. s) , 7.14 (1H, m), 7.42 (1H, m), 7.52 (1H, m) 5 Example 57 7 b -[ (Z)-2- (5-Amino-l,2,4-thiadiazo-l-3-yl)-2- (1- carboxy-l-methylethoxyimino) acetamido]-3-{ [7-guanidino- l-methyl-5- (lH-imidazo [1, 2-b ] pyrazolio) ] methyl ) -3- cephem-4-carboxylate 10 The title compound was obtained from di-tert-butyl {(Z)-[{1-methyl-lH-imidazo[l,2-b]pyrazol-7- yl) aminomethylidene } biscarbamate in the same manner as in Example 43. 1H-NMR(D2O) 1.51 (6H, s), 3.40 (2H, m), 3.85 (3H, s), 15 5.15-5.30 (3H, m) , 5.83 (1H, d, J=4.8Hz), 7.49 (1H, d, J=2.2Hz), 8.02 (1H, d, J=2.2Hz), 8.27 (1H, d, J=1.0Hz) IR(KBr) 3400, 3392, 1770, 1672, 1606, 1531 cm-1 preparation 103 tert-Butyl {3-[ {1-methyl-lH-imidazo [1 , 2-b]pyrzol- 20 7-yl)amino]-3-oxopropyl)carbamate The title compound was obtained from l-methyl-lH- imidazol[1,2-b]pyrazol-7-amine sulfate and 3-[(tert- butoxycarbonyl)amino]propanoic acid in the same manner as in Preparation 70 25 1H-NMR(DMSO-d6) 1.43 (9H, s), 2.61 (2H, m) , 3.49 (2H, m), 3.65 (3H, s), 7.22 (1H, m), 7.26 (1H, m), 7.44 (1H, Example 58 3-|{7-[(3-Aminopropanoyl)amino]-1-methy1-5-(1H- 30 imidazo[1,2-b]pyrazolio) }methyl)-7-[(Z) -2-(5-amino- l,2,4-thiadiazol-3-yl) -2- (1-carboxy-l- methylethoxyimino) acetamido] -3-cephem-4-carboxylate The title compound was obtained from tert-butyl { 3- [ (1-methyl-lH-imidazo [1 ,2-b)]pyrazol-7-yl) amino] -3- 35 oxopropylcarbamate in the same manner as in Example 43. 1H-NMR(D2O) 1.50 (6H, s), 2.97 (2H, d, J=6.5Hz), 3.36 (2H, d, J=6.5Hz), 3.4 (2H, m), 3.81 (3H, s), 5.15-5.30 (3H, m) , 5.82 (1H, d, J=4.8Hz), 7.44 (1H, d, J=2.2Hz), 120 WO 2004/039814 PCT/JP2003/013684 7.98 (1H, d, J=2.2Hz), 3.11 (1H, d, J=1.0Hz) IR(KBr) 3401, 1770, 1665, 1606, 1525 cm-1 Preparation _l04 To a suspension of phenyl [l-methyl-5- 5 {tritylamino) -lH-pyrazol-4-yl] carbamate (4.6 g) in N,N- dimethylformamide (32 ml) were added triethylamine (1.08 g) and tert-butyl l-piperazinecarboxylate (1.99 g). The reaction mixture was stirred for 3 hours and poured into water. The mixture was extracted with ethyl acetate. 10 and the organic layer was concentrated in vacuo. The residue was purified by column chromatagraphy on silica gel (ethyl acetate ethyl acetateethanol=20:1) to give tert-butyl 4- ( { [1-methyl-5- (tritylamino) -1H- pyrazol-4-yl ] amino } carbonyl) -1-piperazinecarboxylate 15 (4.7 g). 1H-NMR(CDCl3) 1.46 (9H, s) , 2.90 (3H, s), 3.05-3.25 (4H, m) , 3.30-3.45 (4H, m) , 4.76 (1H, brs), 5.34 (1H, brs), 7.10-7.30 (l6H, m) Example_59 20 To a solution of 4-methoxybenzyl 7- [ (Z) -2- (5- amino-1, 2 , 4-thiadiazol-3-yl) -2- (l-tert-butoxycarbonyl.-l- methylethoxyimino) acetamido] -3-chloromethyl-3-cephem-4- carboxylate (2 g) in N,N-dimethylformamide (6 ml) was added 1, 3-bis (trimethylsilyl) urea (3 g) , and the 25 reaction mixture was stirred for 30 minutes. Potassium iodide (680 mg) was added to this solution, and the mixture was stirred for 30 minutes, tert-Butyl 4-({1- methyl—5-(tritylamino)-lH-pyrazol-4-yl)amino}carbonyl)- 1-piperazinecarboxylate (2 g) was added to this solution 30 The reaction mixture was stirred at 25oC for 23 hours nd poured into a mixture of ethyl acetate-water-20% queous sodium chloride solution. The organic layer was ashed with a mixture of 10% aqueous sodium thiosulfate olution and 20% aqueous sodium chloride solution. The 35 organic layer was washed successively with 10% aqueous sodium trifluoroacetate solution twice and 20% aqueous sodium chloride solution. The organic layer was concentrated in vacuo to a volume of approximately 10 ml 121 WO 2004/039814 PCT/JP2003/013684 The concentrate was added to diisopropyl ether, and the suspension was stirred for 1 hour, The resulting solid was collected by filtration and dried. The solid was dissolved in dichlromethane (6 ml) . 5 To this solution was added anisole (2 ml) and trifluoroacetic acid (6 ml). The reaction mixture was stirred for 4 hours and poured into diisopropyl ether. The resulting solid was collected by filtration and dried. This solid was purified by preparative HPLC 10 utilizing ODS column. The eluate containing a desired product was concentrated in vacuo. The concentrate was adjusted to about pH 1 with concentrated hydrochloric acid and chromatogaphed on Diaion® HP—20 (Mitsubishi Chemical Corporation) eluting with 20% aqueous 2- 15 propanol. The eluate was concentrated in vacuo, and 2H sulfuric acid was added. The mixture was lyophilizedL to give 3- ( (3-amino-2-methyl-4- [ (1- piperazinylcarbonyl) amino] -1-pyrasolio )methyl) -7- [ (Z) - 2- (5-amino-l ,2 , 4-thiadiazol-3-yl) -2- (1-carboxy-l- 20 methylethoxyimino)acetamido]-3-cephhem-4-carboxylic acid hydrogen sulfate (679 mg) . 1H-NMR(D2O) 1.60 (6H, s) , 3.20 (2H, d, J=17.7Hz), 3.25- 3.45 (4H, m) , 3.45 (1H, d, J=17.7Hz) 3.72 (3H, m) . 3.75- 3.85 (4H, m), 5.00 (1H, d, J=15.7Hz), 5.24 (1H, d, 25 J=15.7HZ), 5.25 (1H, d, J=4.8Hz), 5.86(1H, d, J=4.8Hz) 7.89 (1H, s) This application is based on application No. 2002952355 filed in Australia, on October 30, 2002, and 30 application No, 2003904813 filed in Australia on September 4, 2003, the content of which is incorporated hereinto by reference. 122 WO 2004/039814 PCT/JP2003/013684 wherein 6 R1 is lower alkyl, hydroxy(lower)alkyl or halo(lower)alkyl, and R2 is hydrogen or amino protecting group, or R1 and R2 are bonded together and form lower alkylene Or lower alkenylene; 10 R3 is hydrogen or lower alkyl; R4 is 15 wherein X is O or NH, R7 is hydrogen, lower alkyl or amino protecting group. R8 is hydrogen or hydroxy, protected amino, guanidino, protected guanidino or saturated 3-to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or 25 protected amino, k, m, n and q are independently 0 or 1, and 123 WO 2004/039814 PCT/JP2003/013684 p is 0, 1, 2 or 3; R5 is carboxy or protected carboxy; and R5 is amino or protected amino, or a pharmaceutically acceptable salt thereof. 5 2. The compound of claim 1 wherein R1 is lower alkyl or hydroxy (lower) alkyl , and R2 is hydrogen or amino protecting group, or R1 and R2 are bonded together and form lower alkylene; 10 R3 is hydrogen; A is wherein X is O or NH; R7 is hydrogen or amino protecting group; 15 R3 is amino or protected amino; and p is 0 , 1 or . or a pharmaceutically acceptable salt thereof. 3. Ths compound of claim 2 wherein R3 is hydrogen, or a 20 pharmaceutically acceptable salt thereof. 4. The compound of claim 1 wherein R1 is lower alkyl, hydroxy(lower)alky1 or halo (lower) alkyl, and 25 R2 is hydrogen, aryl(lower)alkyl or acyl, or R1 and R2are bonded together and form lower alkylene or lower alkenylene; R5 is carboxy or esterified carboxy; R6 is amino or acylamino; 30 R7 is hydrogen, lower alkyl or acyl; and R8 is amino, mono or di(lower)alky1 amino, acylamino, guanidino, acylguanidino or saturated 3- to 8- membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or 35 acylamino, or a pharmaceutically acceptable salt thereof. 124 WO 2004/039814 PCT/JP2003/013684 5. The compound of claim 4 wherein R1 is lower alkyl or hydroxy(lower)alkyl, and R2 is hydrogen, aryl(lower)alkyl or acyl, or R1 and R2 are bonded together and form lower alkylene; 5 R5 is carboxy or esterified carboxy; R6 is amino acylamino; R7 is hydrogen or acyl; and R8 is amino or acylamino. or a pharmaceutically acceptable salt therof. 10 6. The compound of claim 5 wherein R1 is alkyl or hydroxy (lower) alkyl, and R2 is hydrogen, aryl(lower)alkyl, lower alkanoyl or lower alkoxycarbonyl, or 15 R1 and R2 are bonded together and form lewer alkylene; R5 is carboxy or lower alkoxycarbonyl; R6 is amino lower alkanoylamino or lower alkoxycarbonylamino; R1 is hydrogen, lower alkanoyl or lower alkoxycarbonyl; 20 and R5 is amino, lower alkanoylamino or lower alkoxycarbonylamino, or a pharmaceutically acceptable salt thereof. 25 7. The compound of claim 6 wherein Rl is lower alkyl or hydroxy(lower)alkyl, and R2 is hydrogern, or R1 and R2 are bonded together and form lower alkylene; R5 is carboxy; 30 R6 is amino; R7 is hydrogen or lower alkanoyl; and R9 is amino, or a pharmaceutically acceptable salt thereof. 35 8. The compound of claim 1 wherein R4 is selected from the group consisting of —NH—A-f NH)—(CH2)—(CH2) p-R14 125 WO 2004/039814 PCT/JP2003/013684 5 wherein R7, A, m, p and q are each as defined in claim 1, R14 is amino, mono or di (lower)alkylamino or protected amino , R15 is guanidino or protected guanidino, and 10 R16 is saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or protected amino, or a pharmaceutically acceptable salt thereof. 15 9. The compound of claim 1 wherein R4 is selected from the group consisting of 126 WO 2004/039814 PCT/JP2003/013684 wherein 5 p is 0, 1 or 2, q is 0 or 1, R7 is hydrogen or amino protecting group, and R9 is amino or protected amino, or a pharmaceutically acceptable salt thereof. 10 10. The compound of claim 9 wherein R7 is hydrogen, lower alkanoyl or lower alkoxycarbonyl; and R9 is amino, lower alkanoylamino or lower 15 a1koxy carbonylamino , or a pharmaceutocally acceptable salt thereof. 11. The compound of claim 10 wherein R7 is hydrogen or lower alkanoyl; and 20 R9 is amino, or a pharmaceutically acceptable salt thereof. 12. A process for preparing a compound of the formula [I]: 127 WO 2004/039814 PCT/JP2003/013684 Wherein Rl is lower alkyl, hydroxy (lower) alkyl or halo(lower)alkyl, and R2 is hydrogen or amino protecting group, or 5 R1 and R2 are bonded together and form lower alkylene or lower alkenylene; R3 is hydrogen or lower alkyl; R4 is wherein is 0 or NH, R7 is hydrogen, lower alkyl or aminc protecting 15 group, R8 is hydrogen or hydroxy, R9 is amino, mono or di(lower)alkylamino , protected amino, guanidino, protected guanidino or saturated 3- to 8-membered 20 heterocyclic group containing 1 to 4 nitrogen atoms optionally substituted by amino or protected amino, k, m, n and q are independently 0 or 1, and p is 0, 1, 2 or 3; 25 R5 is carboxy or protected carboxy; and R6 is amino or protected amino, or a salt thereof, which comprises (1) reacting a compound of the formula. [II] : 128 WO 2004/039814 PCT/JP2003/013684 wherein R1, R2, R3 and R4 are each as defined above, or its reactive derivative at the amino group, or a salt 5 thereof with a compound of the formula [III]: wherein R5 and R6 are each as defined above, or its reactive derivative at the carboxy group, or a salt thereof to give a compound of the formula [I] : wherein R1, R2, R3, R4, R5 and R6 are each as defined0 above, or a salt thereof, or (2) subjecting a compound of the formula [Ia]: 129 WO 2004/039814 PCT/JP2003/013684 wherein R1 , R2, R3, R4 , R5 , R6 , R7 ,R8 A, k, m, n, p and q are each as defined above, and R9a is protected amino, protected guanidino or saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitiogen atoms 5 substituted by protected amino, or a salt thereof to elimination reaction of the amino protecting group to give a compound of the formula [Ib] : 10 wherein R1 , R2, R3, R5 , R6, R7 , R8 , A, k, m, n, p and q are each as defined above, and R9b is amino, guanidino or saturated 3- to 8-membered heterocyclic group containing 1 to 4 nitrogen atoms substituted by amino, or a salt thereof, or 15 (3) reacting a compound of the formula [VI] : wherein R5and R6 are each as defined above, R10 is protected carboxy, and Y is a leaving group, or a salt thereof with a compound of the formula [VII] : 20 wherein R1 , R2, R3 and R4 are each as defined above, or a salt thereof to give a compound of the formula [VIII]: 130 WO 2004/039814 PCT/JP2003/013684 whrein R1 , R2 , R3 , R4 , R5, R6 and R10 are each as defined above, and Z**** is an anion, or a salt thereof, and subjecting the compound of the formula [VIII] or a salt 5 thereof to elimination reaction of the carboxy protecting group, to give a compound of the formula [I]: wherein R1 , R2 , R3 , R4 R5 and R6 are each as defined above, or a salt thereof. 10 13. A pharmaceutical composition comprising a compound of claim 1 or a pharmaceutically acceptable salt thereof in admixture with a pharmaceutically acceptable carrier. 15 14. A compound of claim 1 on a pharmaceutically acceptable salt thereof for use as a medicament. 15 A compound of claim 1 or a pharmaceutically acceptable salt thereof for use as an antimicrobial 20 agent. 16. Use of a compound of claim 1 or a pharmaceutically acceptable salt thereof for manufacture of a medicament for treating infectious diseases, 131 WO 2004/039814 PCT/JP2003/013684 17. A method for the treatment of infectious diseases which comprising administering a compound of claim 1 or a pharmaceutically acceptable salt thereof to human or 5 animals. 132 The present invention relates to a compound of the formula [I]: wherein R1 is lower alkyl, hydroxy(lower)alkyl or halo (lower)alkyl. and R1 is hydrogen amino protecting group, or R1 and R2 are bonded together and form lower alkylene or lower alkenylene; R3 is hydrogen or lower alkyl: R4 is ; R5 is carbony or protected carboxy; and R6 is amino or protected amino, or a pharmaceutically acceptable salt thereof a process for preparing a compound of the formula [I], and a pharmaceutical composition comprising a compound of the formula [I] in admixture with a pharmaceutically acceptable carrier. |
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| Patent Number | 212088 | |||||||||||||||||||||||||||||||||
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| Indian Patent Application Number | 00936/KOLNP/2005 | |||||||||||||||||||||||||||||||||
| PG Journal Number | 46/2007 | |||||||||||||||||||||||||||||||||
| Publication Date | 16-Nov-2007 | |||||||||||||||||||||||||||||||||
| Grant Date | 15-Nov-2007 | |||||||||||||||||||||||||||||||||
| Date of Filing | 19-May-2005 | |||||||||||||||||||||||||||||||||
| Name of Patentee | ASTELLAS PHARMA INC. | |||||||||||||||||||||||||||||||||
| Applicant Address | 3-11, NIHONBASHI-HONCHO 2-CHOME, CHUO-KU, TOKYO 103-8411, JAPAN. | |||||||||||||||||||||||||||||||||
Inventors:
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| PCT International Classification Number | C07D 501/46 | |||||||||||||||||||||||||||||||||
| PCT International Application Number | PCT/JP2003/013684 | |||||||||||||||||||||||||||||||||
| PCT International Filing date | 2003-10-27 | |||||||||||||||||||||||||||||||||
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