CA1338820C - Intermediates useful for preparing cephem compounds and process for preparing same - Google Patents

Abstract

Compounds of the formula:

Description

- 1 - I 33882~
.

The present invention relates to new cephem compounds and pharmaceutically acceptable salts thereof.
~lore particularly, it relates to new cephem compounds and rhArr--eutically acceptable salts thereof, which have antimicrobial activities, to process for preparation thereo~, to ~h~rr-~utical composition comprising the same, and to a method for treating infectious 1; ce~c~c in human being and animals.
Accordingly, one object of the present invention is to provide the cephem compounds and ph~rr-^eutically acceptable 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 the cephem compounds and salts thereof.
A further ob~ect of the present invention is to provide ~h~rr~ceutical composition comprising, as an active ingredient, said cephem compounds or their ~h;~rr~-~eutically acceptable salts.~
Still further object of the present invention is to provide a method for treating in~ectious diseases caused by pathogenic microorganisms, which comprises administering said cephem compounds to infected human being or animals, The object cephem compounds of the present invention are novel a~id can be represented by the following general formula [I]:

Rl~ I ZZ COO~) I
R
wherein Rl and R4 are each amino or a protected amino group, R2 is carboxy (lower) alkyl or a protected 2 0 carboxy ( lower) alkyl, R3 i8 lower alkyl, hydroxy (lower) alkyl or a protected hydroxy (lower) alkyl and R5 is hydrogen or lower alkyl.
As to the object compounds [I], the following points are to be noted.
That is, the object compounds [I] include syn isomer, anti isomer and a mixture thereof. Syn isomer means one geometrical isomer havins the partial structure represented by the following formula:
Rl~ CO
S N--O--R
(wherein Rl and R2 are each as defined above) 1 338~20 and anti isomer means the other yeull~eLLical isomer having the partial structure represented by the followlng formula:
l~N ~ C--CO-(wherein Rl and R2 are each as defined above), and all of such geometrical isomers and mixture thereof are included within the scope of tllis invention.
In the present specification and claim, the partial structure of these geometrical isomers and mixture thereof are represented for convenient sake by the following formula:
R~ --CO--S~
o-R2 (wherein Rl and R2 are each as defined above).
Another point to be noted is that the pyrazolio moiety of the compounds [I] can also exist in the tautomeric form, and such tautomeric equilibrium can be represented by the following schemes.
(--)N/~ R = ~R
(A) (B) (wherein R3, R4 and R5 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 claim, however, the object compounds [I] are represented for the convenient sake by one expression of the pyrazolio group of the formula (A) .
The cephem compounds [I] of the present invention can be prepared by processes as illustrated in the followings.
Process 1 H2N~LCH2--N~R + R ~C--COOH
COO(~
R3 o_R2 [II] [III]
or its reactive or its reactive derivative at the derivative at the amino group carboxy group or a salt thereof or a salt thereof S' N o~L CH2 N~ R4 o_R2 C00(~) l 3 R
[I]
or a salt thereof _ 5 - I 3 3 8 8 2 0 Process 2 E~.5 Rl ~ ~ C--CONH~ R4 O~Ra COO (~) NR3 [Ia]
or a salt thereof Elimination reaction of the carboxy otective roup N S R5 15 pr g ~ R~ --CONH~ 4 O-Rb COO~) I 3 [Ib]
or a salt thereof 20Process 3 R ~ C ~N ~ CH --Y ~ N~
o - R2 COOH R3 [VII ] [V]
or a salt thereof or a salt thereof Rl ~ ~--CONH~ R4 5 (~3 R3 [I]
or a salt thereof - 6 ~ ~ 3 3 8 8 2 0 wherein Rl, R2, R3, R4 and R5 are each as defined above, Ra i8 a protected carboxy(lower)alkyl, Rb is carboxy (lower) alkyl, and Y is a leaving group.

In the above and subsequent ~escriptions of this specification, suitable examples of the various definitions are explained in detail as follows:
The term "lower" i5 intended to mean 1 to 6 carbon atom(s), unless otherwise indicated.
Suitable "protective group" in the "protected amino group" may be lower alkanoyl [e.g. formyl, acetyl, propionyl, hexanoyl, pivaloyl, etc. ], 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.], c~rh yl, arcyl [e . g . benzoyl, toluoyl, naphthoyl, etc . ], ar(lower~alkanoyl [e.g. phenylacetyl, phenylpropionyl, etc. ], aryloxycarbonyl [e.g. phenoxycarbonyl, naphthyloxycarbonyl, etc.], aryloxy(lower)alkanoyl [e.g. phenoxyacetyl, phenoxypropionyl, etc. ], arylglyoxyloyl [e. g, phenylglyoxyloyl, naphthylglyoxyloyl, etc. ], ar (lower) alkoxycarbonyl which may have suitable substituent(s) [e.g. benzyloxycarbonyl, phenethyloxy-carbonyl, p-nitrobenzyloxycarbonyl, etc. ], substituted or unsubstituted ar(lower)alkylidene [e.g. benzylidene, hydroxybenzylidene, etc.], ar(lower)alkyl such as mono or di or triphenyl(lower)alkyl [e~g. benzyl, phenethyi, benzhydryl, trityl, etc. ], or the like, in which the preferred one may be lower alkAnoyl or carbamoyl and the more pre~erred one may be Cl-C4alkanoyl or ~Arh yl.
Suitable "lower alkyl" and "lower alkyl moieties"
in the "carboxy(lower)alkyl", "protected carboxy(lower)-1 33882o alkyl", "hydroxy (lower) alkyl" and "protected hydroxy-( lower) alkyl " may be a straight or hr~nch~d one such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, or the like, in which the preferred one may be Cl-C4alkyl.
Suitable "protected carboxy" in the "protected carboxy(lower)alkyl" may be an estor;f;e~ carboxy group, or the like, and concrete examples of the ester moiety in said esteri~ied carboxy group may be the ones such as lower alkyl ester ~e.g. methyl ester, ethyl ester, propyl ester, isopropyl ester, butyl ester, isobutyl ester, tert-butyl ester, pentyl ester, hexyl ester, l-cyclopropylethyl ester, etc. ] which may have suitable substituent(s), ~or example, lower alkanoyloxy(lower)alkyl ester [e.g. acetoxymethyl ester, propionyloxymethyl ester, butyryloxymethyl ester, valeryloxymethyl ester, pivaloyloxymethyl ester, l-acetoxyethyl ester, l-propionyloxyethyl ester, pivaloyloxymethyl ester, 2-propionyloxyethyl ester, hexanoyloxymethyl ester, etc. ], lower alkanesul~onyl (lower) alkyl ester [e . g.
2-mesylethyl ester, etc. ] or mono (or di or tri) halo-(lower) alkyl ester [e.g. 2-iodoetllyl ester, 2,2,2-trichloroethyl ester, etc.]; lower alkenyl ester [e. g. vinyl ester, allyl ester, etc. ];
lower alkynyl ester [e.g. ethynyl ester, propynyl ester, etc.]; ar(lower)alkyl ester which may have suitable substituent(s) [e.g. benzyl ester, 4-methoxybenzyl ester, 4-nitrobenzyl ester, phenethyl ester, trityl ester, benzhydryl ester, bis (methoxyphenyl)methyl ester, 3,4-dimethoxybenzyl ester, 4-hydroxy-3,5-di-tert-butylbenzyl ester, etc.]; aryl ester which may have suitable substituent(s) [e.g. phenyl ester, 4-chlorophenyl ester, tolyl e'ster, 4-tert-butylphenyl ester, xylyl ester, mesityl ester, cumenyl ester, etc. ];
or the like.

Suitable "protected hydroxy" in the "protected hydroxy(lower)alkyl" may be acyloxy group or the like.
Suitable "acyl moiety" in the "acyloxy" may be lower alkanoyl [e.g. formyl, acetyl, propionyl, hexanoyl, pivaloyl, etc., mono(or di or tri)halo(lower)alkanoyl [e.g. chloroacetyl, trifluoroacetyl, etc.], lower alkoxycarbonyl [e.g. methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, tert-pentyloxycarbonyl, hexyloxy-carbonyl, etc. ] or the like.
Suitable "leaving group" may be halogen [e.g.
chlorine , bromine , iodine , etc . ], acyloxy such as sulfonyloxy [e.g. ~n7e~nesulfonyloxy, tosyloxy, mesyloxy, etc. ], lower alkanoyloxy [e.g. acetyloxy, propionyloxy, etc. ], or the like.
Suitable rh~ eutically acceptable salts of the object compounds [I] are conventional non-toxic 6alts and include a metal salt such as an alkali metal salt [e.g. sodium salt, potassium salt, etc.] and an ;llk~l ;
earth metal salt [e.g. calcium salt, magnesium salt, etc.], an ammonium salt, an organic base salt [e.g.
trimethylamine salt, triethylamine salt, pyridine salt, picoline salt, dicyclohexylamine salt, N,N'-dibenzyl-ethylenediamine salt, etc . ], an organic acid salt [e . g.
formate, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate, bc~n7~on~sulfonate~ toluenesulfonate, etc. ], an inorganic acid salt [e.g. hydrochloride, hydrobromide, sulfate, phosphate, etc. ], a salt with an amino acid [e.g. arginine salt, aspartic acid salt, glutamic acid salt, etc. ], and the like.
Preferred embodiments of the object compound [I]
are as f ollows .
Preferred ~o~l~m~nt of Rl is amino, R2 is carboxy (lower) alkyl [more preferably carboxy-(Cl-C4) alkyl, most preferably carbo~cy Lllyl or l-carboxy-l-methylethyl] or an esterified carboxy (lower) -alkyl [more preferably lower alkoxycarbonyl (lower) -alkyl, most preferably lower alkoxycarbonyl (Cl-C4) -alkyl ], R3 is lower alkyl Imore preferably (Cl-C4)alkyl, most preferably methyl] or hydroxy (lower) alkyl rmore preferably hydroxy(Cl-C4)alkyl, most preferably hydroxyethyl], R4 i5 amino, lower alkanoylamino or ureido, R5 is hydrogen or lower alkyl [more preferably (Cl-C4)-alkyl, most preferably methyl].
(to be continued to the next page) ~ - 10 -1 338~20 The processes for preparing the ob; ect u.,ds of the present invention are ~rl~; n~ in detail in the following .
Process 1 The ob~ect u. ~uul~d ~I] or a salt thereof can be pre-pared by reacting a ul.d tII] or its reactive derivative at the amino group or a salt thereo~ with a _ u.ld [ III ] or its reactive derivative at the carboxy group or a salt thereof.
Suitable reactive derivative at the amino group of the Ulld [II] may include Schif ~' s base type imino or its L~u~ - n enamine type isomer formed by the reaction of the _ __ulld [II] with a carbonyl ~ ,_Ulld ' such as aldehyde, ketone or the like; a 8ilyl derivative formed by the reaction of the __ulld [II] with a silyl compound such as bis (trimethylsilyl) acetamide, mono (trimethylsilyl) acetamide [e.g. N- (trimethylsilyl) -acetamide], bis (trimethylsilyl) urea or the like;
~a derivative formed by reaction of the ~ ,_Ull~ [II]
wi h phosphorus trinh1ori~e or phosgere, and the like.
Suitable salts of the ~ __Ull~ [III and its reactive derivative can be refe-red to the ones as exe~olified for the cu~uulld [I].
Suitable reactive derivative at the carboxy group of the cu .~su..d [III] may include 2n acid halide, an acid anhydride, an activated amide, an activated ester, and the like. Suitable ~-Y~mrl~C of the reactive de_ivat_ves may be a~ acid chloride; an acid azide;
a mixed acid anhydride with an acid such as substituted phosphoric acid [e.g. dialkyl~hnsrhnric acid, phenylpllo2~ o ic acid, diphenylphosphoric acid, dibenzylphosphoric acid, halogenated r1~n~L,l ~ ic acid, etc.], dialkylr~nsphnrous acid, sulfurous acid, thiosulfuric acid, sul~uric acid, sulforic acid [e.g.
--. .

methanes~llfonic acid, etc. ], aliphatic carboxylic acid [e.g. acetic acid, propionic acid, butyric acid, is~LuLy~ic acid, pivalic acid, pentanoic acid, isopentanoic acid, 2-ethylbutyric acid, trichloroacetic acid, etc.] or aromatic carboxylic acid [e.g. benzoic acid, etc.]; a ~y L.ical acid anhydride;
an activated amide with imidazole, 4-substituted ;m;~ ol e, dimethylpyrazole, triazole or tetrazole; or an activated ester [e.g. cy~n~ - Lllyl ester, methoxymethyl ester, dimethyl;m;nl -thyl [(C~3)2i~=CE-] ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenyl thioester, p-nitrophenyl thioester, p-cresyl thioester, carb~"~y L1LY1 thioester, pyranyl ester, pyridyl ester, piperidyl ester, 8-quinolyl thioester, etc. ], or an ester with a N-hydroxy compound [e.g. N,N-dimethyl-hydroxylamine, l-hydroxy-2- (lH) -pyridone, N-hydroxy-5llrr;n;m;tl~, N--hydroxyphth~l ;m;~e, 1--hydroxy-l~I-benzotriazole, etc. ], and the like. These reactive derivatives can optionally be selected from them accord-ing to the kind of the ~ __ L..d [III] to be used.
Suitable salts of the compound [III] and its reactive derivative can be ref erred to the ones as exemplified for the compound [I].
The reaction is usually carried out in a convPnt; rn;l 1 solvent such as water, alcohol [e.g. methanol, ethanol, etc.], acetone, dioxane, acetonitrile, chloroform, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide, pyridine or any other organic solvent which does not adversely ;nf1~1~nre the reaction. These conventional solvent may also be used in a mixture with water., In this reaction, when the c ~ l [III] is used in a free acid form or its salt form, the reaction is - 12 - l ~38820 preferably carried out in the presence of a conventional cnnfl~n~;ng agent such as N,N'-dicyclohexylcarbodiimide;
N-cyclohexyl-N ' -mor~hnl; nnethylcarbodiimide;
N-cyclohexyl-N ' - ( 4 -diethylaminocyclohexyl ) r :~ rhofl;; m; fl.~;
N,N'-diethylcarbodiimide, N,N'-diisopropyl~rho~l; ;m;fl~;
N-ethyl-N'-(3-dimethylaminopropyl)carbofl;;m;A~; N,N'-carbonylb i s- ( 2 -me thy l ; m ; fl ~ ~ o le ); rc n t Lhyleneketene-N-cyclohexylimine; diphenylketene-N-cyclohexylimine;
ethoxyacetylene; l-alkoxy-l-chloroethylene; trialkyl phosphite; ethyl polyphosphate; isopropyl polyphosphate;
phosphorus oxychloride ~phosphoryl chloride);
phosphorus trichloride; thionyl nhloriflo; oxalyl chloride;
lower alkyl haloformate [e.g. ethyl chloroformate, isopropyl chloroformate, etc. ]; triphenylphosphine;
2-ethyl-7-hydroxyb~n7; COY l701ium salt; 2-ethyl-5-(m-s~ll fsrh~-nyl); cny~ol ;llm hydroxide intramolecular salt;
1 - ( p -chlo rnh ~n 7~ n 0 ~ ul f ony loxy ) - 6 - chloro - lH-ben zotr ia zo le;
so-called V~ l ! ; er reagent prepared by the reaction of N,N-dimethylfc rm~m; rl,~ with thion~l chloride, phosgene, trichloromethyl chloroformate, phosphorus oxychloride, etc.; or the like.
The reaction may also be carried out in the presence of an inorganic or organic base such as an alkali metal blc~rhnnate, tri(lower)alkylamine, pyridine, N- (lower) alkylmorpholine, N,N-di (lower) alkylbenzylamine, or the like.
The reaction temperature is not critical, and the reaction is usually carried out under cooling to warming.
Process 2 The object compound [Ib] or a salt thereof can be prepared by subjecting a compound [Ia] or a salt thereof to elimination reaction of the carboxy protective group.
This reaction is carried out in accordance with a conventional method such as hydrolysis, reduction or the like .

~ ~ 13 - 1 338820 The hydrolysis is preferably carried out in the presence of a base or an acid including Lewis acid.
Suitable base may include an inorganic base and an organic base such as an alkali metal [e . g. sodium, potassium, etc . ], an A 1 kA l; nP earth metal [e . g . magnesium, calcium, etc. ], the hydroxide or carbonate or bicarbonate thereoE, trialkylamine [e.g. trimethylamine, triethylamine, etc.], picoline, 1,5-~; A7AhiCyclo[ 4.3.O]non-5-ene, 1,4--,1; A7Ah; Pyclo [2 . 2 .2] octane, 1, 8--~1; A7Ah; cyclo [5 . 4 . O]
undec-7-ene, or the like.
Suitable acid may include an organic acid [e . g. 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, etc. ] .
The elimination using Lewis acid such as trih~ A~ etic acid [e.g. trichloroacetic acid, trifluoroacetic acid, etc. ] or the like is preferably carried out in the presence o~ cation trapping agents [e. g. anisole, 2 0 phenol, etc . ] .
The reaction is usually carried out in a solvent such as water, an alcohol [e.g. methanol, ethanol, etc.], methylene chloride, tetrahydrofuran, a mixture thereof or any other solvent which does not adversely influence the reaction. A liquid base or acid can be also used as the solvent. The reaction temperature is not critical and the reaction is usually carried out under cooling to warming.
The reduction method applicable for the elimination reaction may include chemical reduction and catalytic reduction .
Suitable reducing agents to be uged in l~h~m; CAl reduction are a combination of metal [e . g . tin, zinc, iron, etc. ] or metallic compound [e. g. chromium chloride, chromium acetate, etc. ] and an organic or inorganic ~ ~ 14 ~ 1 3 3 8 8 2 0 acid [e.g. formic acid, acetic acid, propionic acid, trifluoroacetic acid, p-toluenesul~onic acid, hydrochloric acid, hydrobromic acid, etc.].
Suitable catalysts to be used in catalytic reduction are conventional ones such as platinum catalysts [e . g.
p~atinum plate, spongy platinum, platinum black, colloidal platinu~., platinum oxide, platinum wire, etc. ], palladium catalysts [e.g. spongy r~llA~ m, p~llA~ m black, palladium oxide, palladium on carbon, colloidal pAllA~;llm, rAllA~ m on barium sulfate, rAllA~ m on barium carbonate , etc . ], nickel catalysts [e . g . reduced nickel, nickel oxide, ~aney 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.
The reduction is usually carried out in a conven-tional 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 abovementioned acids to be used in rhr~m; CAl reduction are in li~uid, 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, dioxane, tetrahydrofuran, etc., or a mixture thereof.
The reaction temperature of this redl-rt; on is not critical and the reaction is usually carried out under cooling to warming.

.

~ - 15 - t ~3 8 8 2 0 Process 3 The object compound [I] or a salt thereof can be prepared by reactinq a compound [VII] or a salt thereof with a compound [V] or a salt thereof.
Suitable salts of the compounds [V] and [VII] can be referred to the ones as exempliied for the compound [I] .
The present reaction may be carried out in a solvent such as water, phosphate buffer, acetone, chloroform, acetonitrile, nitr~hpn 7PnP, methylene chloride, ethylene chloride, for~-m;~lP, N,N-dimethyl-~Qrm-m;~lP~ methanol, ethanol, diethyl ether, tetrahydro-furan, dimethyl sulfoxide, or any other organic solvent which does not adversely affect the reaction, preferably in ones having strong polarities. Among the solvents, hydrophilic solvents may be used in a mixture with water.
When the compound [V] is in liquid, it can also be used as a solvent. The reaction is preferably conducted in the presence of a base, for example, inorganic base such as alkali metal hydroxide, alkali metal carbonate, alkali metal h; c~h~n~te, organic base such as trialkylamine, and the like. The reaction temperature is not critical, and the reaction is usually carried out at ambient temperature, under warming or under heating.
The present reaction is preferably carried out in the presence of alkali metal halide [e . g . sodium iodide, potassium iodide, etc.], alkali metal thiocyanate [e.g.
sodium thiocyanate, potassium thiocyanate, etc. ] or the like .
The starting compound [II ] is novel and can be prepared by the f ollowing processes A and B .

~ 1 338820 - l5A -According to another aspect of the invention there is provided:
1. A compound of the formula:

E~2N~ ~
o~ N~CH2_N
COO (3 R3 wherein R3 is lower alkyI, hydroxy(lower)alkyl or a protected hydroxy (lower) alkyl, R is arr~ino or a protected amino group, and R5 is hydrogen or lower alkyl, and a salt thereof.

According to yet another aspect of the invention there is provided a process for preparing the above compound which comprises:
(1) subjecting a compound of the ~ormula:

~ ~L 2 2 5 R7 Ra wherein R4 and R5 are each as defined above, Ra is hydroxy (lower) alkyl or a protected 3 hydroxy (lower) alky1, R is a protected amino group, R7 is a prote!cted carboxy group, and X~)is an anion, 1~ - l5s - 1 33882 or a fialt thereof to elimination reaction o~
the amino protective group in R6 and the carboxy protective group in R7 to give a compound oi the ~ormula:

H2N ¦ I'S' (~3~R
o,J--N ~ C~ 2--N `

COO (~) Ra wherein Ra ~ R4 and R5 are each afi defined above, or a fialt thereof, or (2) subjecting a compound of the ~ormula:

H2N~ 1 ~3 /~ R4 o ~ ~ CH2 N~N~ a COO(~) la w~lerel~ a and R are each as defined above, and Ra is a protected amino group, or a salt thereof to elimination reaction of the amino protective group to give a compound o~ the ~ormul~l:

~LCH2~ NH2 COO~

4~ i ~38820 -- 15 c --wherein Ra and RS are eaeh as defined above, or a salt thereo~, or (3) subjecting a compound of t~le formula:
Ra~L `~
wherein R4, R5, R7 and l{~are each as de~ined above, Rb i9 lower alk}~l, and Ra is hydroxyberlzyl;~ne~m;n.., or a salt thereof to elimination reaction of the amino protective c;roup in Ra and the carboxy protective group in R7 to give a eompound of the formula:
2~ R5 ~L CH2--N~
Coo(~) ~
wherein Rb, R4 and R5 are each as defined above, or a salt thereof, or (4) subjecting a compound of the formula:
~, ~ - 15D - t 338820 H2N~

wherein Ra is hydrogen or Illethyl, or a salt thereof . to de~ormylation to give a aompound of the formula:

H2N ~S~ H ,r--(Ra o~L ~c 2 N~ N~NH2 COO~ CE~3 wherein R5 is as defined above, or a salt thereof.

- 16 - ~33882 Proce6s A
R6~_~S~
[IV]
or a salt thereof N~S or a 6alt thereof ~ CH2 2$

[VI ]
or a salt thereo:E

112N l,S `1 (3~R
,~ N~ C~2--N~
COO ~ R3 [II]
or a 6alt thereof ~ 1 3~8820 Process B ~ R5 ~ CH2~N~a COO(~) R3 [IIa]
or a salt thereof v R5 ~L CH2--COO (~) R3 [IIb]
or a salt thereof wherein R3, R4, R5 and Y are each as defined above, R6 iB a protected amino group, R7 is a protected carboxy group, X(~3i8 an anion, and Ra is a protected amino group.
Suitable "protective group " in the "protected amino group" for R6 can be referred to the ones as exemplified before and the preferred one may be substituted or unsubstituted ar ( lower) alkylidene (e . g ., benzylidene, hydroxybenzylidene , etc. ), lower alkoxycarbonyl (e . g. , methoxycarbonyl, ethoxycarbonyl, tert-butoxycarbonyl, tert-pentyloxycarbonyl, hexyloxycarbonyl, etc. ) .
Suitable "protected carboxy grOUp" for R7 can be referred to the ones as exemplified before and the - 18 - ~ ~ ~i 8 8 2 0 preferred one may be ar ( lower) alkoxycarbonyl which may have suitable substituent (s) (e . g. benzhydryloxycarbonyl, etc.) .
Suitable "anion" may be formate, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate, toluenesulfonate, chloride, bromide, iodide, sulfate, phosphate, or the like.
Processes A and B for the preparation of the starting compound [II~ is explained in detail in the following.
Process A - (~
The compound [VI ] or a salt thereof can be prepared by reacting the compound [IV] or a salt thereof with the compound [V] or a salt thereof. This reaction can be carried out in a similar manner to that of the aforementioned Process 3, and therefore the reagents to be used and the reaction conditions (e.g., solvent, reaction temperature, etc. ) can be referred to those of the Process 3.
Anion ~) may be the one derived from a leaving group Y and may be the other one converted therefrom by a conventional method.
Process A - (~
The compound [II] or a salt thereof can be prepared by subjecting the compound [VI] or a salt thereof to elimination reaction of the amino protective group in R6 and the carboxy protective group in R7.
This reaction is carried out in ~cQr~nce with a conventional method such as hydrolysis or the like.
The hydrolysis is preferably carried out in the presence of a base or an acid including Lewis acid.
Suitable base may include an inorganic base and an organic base such as an alkal!i metal [e . g. sodium, potassium, etc.], an Alk~l;n~ earth metal [e.g.

~ T 1 9 magnesium, calcium, etc. ], the hydroxide or carbonate or hic~rho~te thereof, trialkylamine [e.g. trimethyl-amine, triethylamine, etc.], picoline, 1,5-fl;~h;cyclo-[ 4 . 3 . 0 ] non-5 -ene, 1, 4 -diazabicyclo [ 2 . 2 . 2 ] octane, 1, 8 -fl;zl7~hi~-yclo[5.4.0]undec-7-ene, or the like.
Suitable acid may include an organic acid [e.g. formic acid, aceti-~c acid, propionic acid, trichloroacetic acid, trifluoroacetic acid, etc. ] and an inorganic acid [e.g.
hydrochloric acid, hydrobromic acid, sulfuric acid, hydrogen chloride, hydrogen bromide, etc. ] .
The elimination using Lewis acid such as tri h~ r-etic acid [e.g. trichloroacetic acid, trifluoroacetic acid, etc. ] or the like is pre~erably 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 , an alcohol [e . g. methanol , ethanol, etc . ], methylene rhlrlrifl~ tetrahydrofuran, a mixture thereof or any other solvent which does not adversely influence the reaction. A liquid base or acid can be also used as the solvent. The reaction temperature is not critical and the reaction is usually carried out under cooling to warming.
The present invention includes within the scope of the invention the case that protected amino in R4 is transformed into amino during this reaction.
Process B
The compound [IIb] or a salt thereof can be prepared by subjecting the compound [IIa] to elimination reaction of the amino protective group in Ra. This reaction can be carried out in a similar manner to that of the aforementioned Process A - (~), and therefore the reagents to be used and the réaction conditions (e . g., solvent, reaction temperature, etc. ) can be referred to those of the Process A - (~) .

The present invention includes within the scope of the invention the case that protected l~ydroxy (lower) alkyl in R3 is transformed into hydroxy~lower)alkyl during this reaction.

me object compounds [I] and rh~rm-~eutically acceptable salts thereof are novel and exhibit high antimicrobial activity, inhibiting the growth of a wide variety of pathogenic microorganisms ; nc 1~ ; ng Gram-positive and Gram-negative microorganisms and are useful as antimicrobial agents.
Now in oraer to show the utility of the object compounds [I], the test data on MIC (minimal inhibitory concentration) of representative compounds [I] of this invention are shown in the following.
Test method:
In vitro antibacterial activity wa6 det~r~; n~-d by the two-fold agar-plate dilution method as described below.
One loopful of an overnight culture of each test strain in Trypticase-soy broth (106 viable cells per ml) was streaked on heart infusion agar (~I-agar) containing graded concentrations of representative test compound, and the minimal inhibitory concentration (MIC) was expressed in terms of ,ug/mQ after incubation at 37C
for 20 hours.
Test compounds:
(1) 7~--[2-(5-Amino-1,2,4--th;~ 7Ol-3--yl)-2-carboxymethoxy;m;noacetamido]-3-(3-amino-2-methyl-l-pyrazolio) methyl-3-cephem-4-carboxylate ~syn isomer) (hereinafter referred to as Compound A).
(2) 7~--[2--(5--A~ino--1,2,4--th;~ 7~1--3--yl)--2--carboxymethoxyiminoacetamido] -3- (3-formamido-2-methyl-l-pyrazolio) methyl-3-cephem-4-carboxylate ~syn isomer) (hereinafter referred to as Compound B).
(3) 7~--[2--(5--Amino--1,2,4--th;~-l;A7ol--3--yl)--2--(l--carboxy-l-methylethoxyimino) acetamido] -3- (3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate (syn isomer) (hereinafter referred to as Compound C).
(to be continued to the next page) ,~ 1 33882û
Test results:
MIC (,ug/ml) 5 Tes t Test compounds strain A B C
vu P . lgarls 0 . 20 o . 20 o .10 For therapeutic administration, the object ~ ~ u..ds [Il and ph~rr^^e~ltically acceptable salts thereof of the present invention are used in the form of conven-tional rh~rr~Aeutical preparation which cc~ntain6 said __ u..d as an active ingredient, in admixture with LAh;lrr~Aeutically acceptable carriers such as an organic or; nnr~Rn; ~- solid or liyuid excipient which is suitable for oral, parenteral and external ~dministration.
The phArr-~Aeutical preparations may be in solid form such as tablet, granule, powder, capsule, or li.~uid form such as solution, suspension, syrup, emulsion, lA-A~r~qe and the like.
If needed, there may be; n~Aln~e~l in the above preparations ~ ry substances, stabilizing agents, wetting agents and other commonly used additives such as lactose, citric acid, tartaric acid, stearic acid, magnesium stearate, terra alba, sucrose, corn starch, talc, gelatin, agar, pectin, peanut oil, olive oil, cacao butter, ethylene glycol, and the like.
While the dosage of the compound [I] may vary from and also depend upon the age, conditions of the patient, a kind of diseases, a kind of the _ ~ .d [I~ to be applied, etc. In general, amounts between 1 mg and about 4,000 mg or even more per day may be administered to a patient. An average single dQse of about 50 mg, 100 mg, 250 mg, 500 mg, 1000 mg, 2000 mg of the ob~ect compounds [I] of the present invention may be used in treating diseases infected by pathogenic microorganisms.
The following Preparations and Examples are given for the purpose of illustrating the present invention in more detail.
Preparation 1 A mixture of acetic anhydride (38 . 86 ml) and formic acid (15.54 ml) was stirred at 45C for 45 minutes. To this mixture was added 5-amino-1-methylpyrazole (10 g) under ice-cooling, and the reaction mixture was stirred at the same temperature for 10 minutes. The resultant mixture was poured into a mixture of water and ethyl acetate, and the resultant solution was ad~usted to p~I 8 with potassium carbonate.
The organic layer was separated, and the aqueous layer was extracted with ethyl acetate six times. The organic layers were combined, dried over magnesium sulfate, and evaporated in vacuo to give 5-form:lm;rln-?-methylpyrazole (12. 88 g) .
mp:71-73C
25 IR (Nujol)*: 3300, 3200, 1705, 1590 cm (CDC13, ~) : 3.69 and 3.74 (3FI, each s), 6.04 and 6.23 (l~I, each d, J=3~z), 7.34 (l~I, s), 8.21 (1~, s) 3 0 Preparation 2 The following compounds were obtained according to a similar manner to that of Preparation 1.
(1) 4-F~ m; ~ methylpyrazole 35 mp: 44-45C
* Trademark - _ -- 24 --IR (Nujol): 3250, 1665, 1585 cm ~
NMR (CDC13, ~:3.83 (3~, s), 7.33 (11~, s), 7.83 tlE~, s), 8.17 (1~, s) (2) 5-Fn--r~mi~ln-1,4-dimethylpyrazole IR tNujol): 3200, 1665, 1585 cm 1 N~SR (CDC13, ~) : 1.90 and 1.98 (3E, each s), 3.64 and 3.72 (3~, each s), 7.29 and 7.31 (1~, each s), 8.10 (lE, broad 5) 8.33 and 9.03 (1~1, each s).
Preparation 3 To a mixture of benzhydryl 73-te:L L-~uLu~y-.ia ~ bc/~.ylamino-3-chloL U-yl-3 - cephem-4-carboxylate (15 g) and sodium iodide (4 . 37 g) in acetone (15 ml) was added 5-fn~-m; ~n_ l--methylpyrazole (15 g) at ambient temperature, After being stirred for 40 hours at the same temperature, the mixture was poured into a mixture of water &nd ethyl l~cetate . The organic layer was separatea and washed with water, aqueous sodium chlnr;rie solution, and dried over magnesium sulfate.
The sol~lti nn was evaporated in vacuo to give benzhydryl 73-te ~-~uLo~y~rbonylamino-3- (3--fnr -=m; rio-2-methyl-l-pyrazolio)methyl-3-cephem-4-carboxylate iodide (20.95 g).
IR (Nujol): 1780, 1710, 1580 cm 1 NrqR (D~SO-d6, ~) : 1.40 (9~I, s), 3.41 (2}~, broad s), 3 . 65 (3~I, s), 5 .12 (1~1, d, 3=5Ez), 5.36 (2}I, broad s), 5.57 (lE, dd, J=8~z and 5Elz), 6.88 (l}I, s), 6.89 (1~, m), 7.10-7.48 (10~, m), 7.83 (lI~, d, J=8~1z), 8.24 (lEI, d, J=3Ez), 8.45 (l~I, s) Preparation 4 The following ~ ds wére o}~tained E~cnr~; "s to a similar manner to that of PreparatiOn 3.

-25 ~ l 338820 (1) Benzhydryl 7~-tert-buLu~yc~lrbonylamino-3- (4-fo~qm; ~- 2-methyl-1-pyrazolio ) methyl-3-cephem-4-carboxylate iodide IR (Nu~ol) : 1785, 1720, 1605 cm NMR (DMSO-d6, ~) : 1.39 (9E~, s), 3.42 (2H, broad s), 3.77 (3~, s), 5.11 (lE, d, J=5Rz), 5.41 (2H, broad 8), 5.60 (lE, dd, J-8~z and 5EIz), 6.89 (l~, s), 7.18-7.52 (lOEI, m), 7.96 (lH, d, J=8~z), 8.25 (lE~, s), 8.51 (lH, s), 8.57 (l~, s) .
(2) Benzhydryl 7~-tert-butoxycasbonylamino-3- (3-for~ m;~o-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylate iodide.
IR (Nujol): 3300, 1780, 1705 cm 1 NMR (DMSO--d6, ô) : 1.42 (9EI, s), 1.98 (3~I, s), 3.45 (2~, broad 8), 3.63 (3E, s), 5 . 19 ( lH , d , J=5~z ), 5 . 4 0 ( 2~I , broad s ), 5.61 (1~, dd, J=5E~z and 8Rz), 6.95 (lE, s), 7.21-7.58 (lOE~, m), 8.00 (lF~, d, J~8~z), 8.21 (lE, s), 8.43 (1~, s) Preparation 5 To a solution of benzhydryl 7~-tert-butoxycarbonylamino-3- (3-for~-m; ~in-2-methyl-1-pyrazolio)met~yl-3-cephem-4-carboxylate iodide (20.9 g) and anisole (20 ml) in me_hylene chloride (40 ml) was added dropwise t~; f~ roacetic acid (40 ml) under ice-cooling. After being stirred for l. 5 hours at ambient temperature, the mixture was poured into a mixture of diisopropyl ether (300 ml) and ethyl acetate (300 ml). The resultant precipitate was col-lected by filtration to give, di.(trifluoroacetic acid) salt of 7~-amino-3- (3-forr-m;~-2-methyl-1-pyrazolio)-35 methyl-3-cephem-4-carboxyla~e - (16.20 g).

~ 26 ~ 1 338820 IR (Nujol): 3350, 1770, 1660 cm ~MR (DMSO--d6, ~) : 3.45 (2E, s), 3.87 (3E, 8), 5.18 (2~I, s), 5.47 (2E, s), 6.95 (lH, d, J=3~z), 8.33 (lE, d, J=3Ez), 8.47 (lH, s).

Preparation 6 The following compounds were obtained according to a similar manner to that of Preparation 5.
(1~ Di~rifluoroacetic acid) salt of 7~-amino--3- (4-form-m; lrl-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxyl ate .
IR (~lu~ol): 3400, 1780, 1660, 1605 cm 1 NMR (DMSO-d6, ~) : 3.51 (2E, broad s~, 4.06 (3E, s), 5.23 (2E~, s), 5.55 (2H, broad s), 8.30 (lE, s), 8.61 (lE, s), 8.67 (lEI, s) .
(2) Di(trifluoroacetic acid) salt of 78-amino-3- (3-for~n~Tn; c1O-2, 4-aimethyl-1-pyrazolio) methyl-3-20 cephem-4-carboxyla~e.
~MR (DMSO-d6, ~) : 2.01 (3E, s), 3.48 (2E, broad sj, 3.83 (3H, s), 5.24 (2E, s), S.50 (2E, broad s), 8.26 (lE, s), 8.41 (lE, s) .
25 Preparation 7 Concentrated hydrochloric acid (0.353 ml) was added to a mixture of di(trifluoroacetic acid) salt of 7~-amino-3-(3-f~rr-mi~---2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate (0 . 565 g) in tetrahydrofuran (3 ml) and methanol (3 ml) at ambient temperature.
after being stirred at the same temperature for 12 hours, the mixture was added dropwise to ethyl acetate (100 ml). The resultant precipitate was collected by filtration to give 7~-amino-3- (3-amino-2-methyl-l-pyrazolio) methyl-3-cephem-4-carboxylate trihydrochloride (292 mg).
.

~ 27 ~ 1 338820 NMR (D~3O-d6, ~ ): 3.31 and 3.56 (2H, ABq, J-- 18 Hz), 3.67 (3H, s), 5.20 (2H, broaa s), 5.29 (2H, broad s), 5.87 (lH, d, J=3Hz), 8.12 (lH, d, J=3Hz) Preparation 8 7~-Pmino-3-(3- mino-2,4-dimethyl-1-pyrazolio)methyl-3-cephem-4-carboxylate trihydrochloride was obtained according to a similar manner to that of Preparation 7.
IR (~ujol): 3350, 1780, 1640 cm 1 NMR (DMSO--d6, ~) : 1.97 (3H, 5), 3.49 (2H, s), 3.74 (3H, s), 5.27 (4H, s), 8.00 (lH, s) .
Example 1 To a solution of di~rifluoroacetic acid) salt of 7~-amino-3- (4-formamido-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate (4. 0 g) and N-(fr;~-U.ylsilyl)acetamide (9.29 g) in tetrahydrofuran (60 ml) was added methanesulfonyl (Z)-2- (5-amino-1,2,i-th;Afl;;~l701-3-yl)-2-ca ~yllL, Lhoxy;minr~cetate (2.07 g) under ice-cooling. The mixture was stirred for 1 hour at ambient temperature and then the resulting mixture was added dropwise to diisopropyl ether. Thus produced precipitate was collected by filtration, dissolved in water and then subjected to column chromaLuy c,p~.y on ma, Lu~oLous non-ionic adsorption resin, Diaion HP-20 (Trademark: manufactured by Mitsubishi Chemical Industries).
The desired product was eluted with 10% a~ueous isopropyl alcohol solution, and then lyo~hi~ fl to give 7~-[2-(5-amino-1,2,4-th;Afl;A7ol-3-yl)-2--ca bu~y -thoxyiminoacetamido]_3_ (4-f ~rr~m; d~-2-methy1-1-pyrazolio)methyl-3-cephem-4-carboxylate (syn isomer 0 . 714 g) .
IR (Nujol): 3300, 1765,'1665, 1605 cm 35 NMR (D20, ~) : 3.18 and 3.51 (2H, AB~, J=1811z), t 3~8820 4.09 (3~, s), 4.63 (2E, s), 5.22 and 5.46 ~2~, ABq, J=15Hz), 5.23 (1~, d, J=53~z ), 5 . 8 6 ( lE , d , J=5E~z ), 8.25 (lH, s), 8.36 (1~, s), 8.43 (lH, s) Example 2 (1) 7~--[2--(5-P,mino-1,2,4--~hi~ 701-3--yl)-2--caLbo~yllLethoxy; m; n^acetamido] -3- (3-fQ~~m; ~
2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) was obtained according to a similar manner to that of Example 1.
IR (Nu~ol): 3300, 1760, 1660, 1580 cm 1 NMR (D20, ~ ) : 3.13 and 3.44 (2~, A~3q, J=18Hz), 3.91 (3E~, s), 4.67 (2E~, s), 5.21 (lE, d, J=5E~z), 5.22 and 5.43 (2E, ABq, J=15E~z), 5.85 (lE, d, J=5Elz), 6.82 and 6.92 (1~, each d, J=3Hz), 8.15 (1~, d, J=3E~z), 8.40 (lX, s).
(2) 7~--~2--(5--Amino--1,2,4--~h;~ 701--3--yl)--2--carboxymethoxy; m; n~ etamido I - 3- ( 3- fo~slm; ~
2, 4-dimethyl- 1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) was obtained according to ~ similar manner to that of Example 1.
IR (Nujol): 3300, 1765, 1660, 1600 cm 1 NMR (D2O, ~) : 2.03 (3~, s), 3.17 and 3.48 (2~I, A3q, J=18Hz), 3.83 (3E, s), 4.70 (2~, s), 5.18 and 5.43 (2~, A~3q, J=15Hz), 5.24 (1~, d, J=5EIz), 5.87 (1~, d, J=5~z), 8.05 (lE~, s), 8.36 (1~, s) Example 3 To a solution of 7~-amino-3- (3-amino-2-methyl-l-pyrazoli~methyl-3-cephem-4-carboxylate trihydrochloride (6.96 g) and N-(trimethylsilyl)acetamide (21.8 g) in ~ - 2~ ~ 338820 tetrahydrofuran (150 ml~ was added methanesulConyl (Z)-2-(5-amino-1,2,4-~h;Ar~;A7ol-3-yl)-2-carboxymethoxyiminoacetate (4 . 86 g) at ~mbient temperature. After being stirred for 30 minutes at the same temperature, the mixture was poured into diethyl ether ( 2 Q ), and the resultant pre-cipitate was collected by filtration. The precipitate was dissolvea in water, and the solution was ad~usted to pH 2 . 0 with aqueous sodium bicArhonAte solution.
The solution was subjected to column chromatography on ma~Lu~Luus non-ionic adsorption resin, Diaion HP-20.
The desired product ~as eluted with 5% isopropyl alcohol solution and lyorh;l;~ecl to give 73-~!-(5-amino-1,2,4-fh;Arl;~7ol-3-yl)-2-ca,Lu~y",_~hoxy;m;nnAr~etamido]-3-(3-amino-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) (5. 20 g) .
IR (Nu~ol): 3300, 1760, 1660, 1590 cm 1 N~R (D20, ~) : 3.06 and 3.33 (2H; ABq, J=18Hz), 3.63 (3H, s), 4.60 (2H, s), 4.93 and 5.21 (2H, A3q, J=15Hz), 5.16 (lH, d, J=5Hz), 5.83 (lX, d, J=5Hz), 5.88 (lH, d, J=3Hz), 7.78 (lH, d, J=3Hz) .
Example 4 To a solution of 7~-amino-3- (3-amino-2,4-dimethyl-l-pyrazolio) methyl-3-cephem-4-carboxylate tri~ydrochloride tl.0 g) and N-(trimethylsilyl)-acetamide (3.03 g) in tetrahyarofuran (20 ml) was added methanesulfonyl (Z)-2-(5-amino-1,2,4-~h;A~;A701-3-yl)-2-CalLù~sy -thoxyi~;n(~r~cetate (0.68 g) at ambient temperature. After being stirred for 1 hour at the same temperature, the mixture was poured into diethyl ether (300 ml) and the resultant precipitate was collected by f iltration . The pre-cipitate was dissolved in water and the solution was ~o ad~usted to pE~ 2. 0 with aqueous sodium bicarbonate solution .
The solution was subjected to a column chromatography on ma.;Lv~,oL.,us non-ionic adsorption resin, Diaion ~P-20. The desired product was eluted with 5~ isopropyl alcohol aqueous solution and lyorh;l;7~d to give 7~-[2-(5-amino-1,2,4-~h;aA;
3-yl) -2-carboxymethoxy;m;nnAcetamido]-3- (3-amino-2, 4-dimethyl-1-pyrazolio) methyl-3-cephem-4-aarboxylate (syn isomer) (0.165 g).
IR (Nu~ol): 3350, 1770, 1660, 1600 cm NMR (D2O, ~) : 1.93 (3~, s), 3.06 and 3.30 (2~, ABq, J=18~z), 3.64 (3E, s), 4.67 (2~, s), 4.88 and 5.19 (2H, ABq, J~15~z), 5 . 18 (1~, d, J=5E~z), 5 . 84 (lE, d, J=5Ez), 7 . 66 (lE, s) .
(to be continued to the next page) .

Preparation 9 To a solution of phosphorus pentachloride (11.11 g) in methylene chloride (167. 8 ml) was added (Z) -2- (5-amino-l, 2, 4-th; ~fl; :1 701-3-yl ) -2- ( l-tert-butoxycarbonyl-l-methylethoxyimino) acetic acid (16.78 g) at -20C.
The resultant mixture was stirred at -20 to -10C for 1. 5 hours, and to the mixture was added dropwise diisopropyl ether (671.2 ml) at -20 to -10C. The mixture was stirred under ice-cooling for 1 hour and the resultant precipitate was collected by filtration to give (Z)-2-(5-amino-1,2,4-~h;Ar~ ol-3-yl)-2-(1-tert-butoxycarbonyl-l-methylethoxyimino ) acetyl rh 1 r r; fl~
hydrochloride (14 . 8 g) .
IR (Nujol): 3430, 3270, 3130, 1815, 1750, 1725, 1640 cm 1 Example 5 To a solution of 73-amino-3-(3-ami~o-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate trihydrorh1 or (2 g) and N-(trimethylsilyl)acetamide (6.28 g) in tetrahydrofuran (40 ml) was added (Z) -2- (5-amino-l,2,4-~h; ~rl; S~7r~1-3-yl) -2- (l-tert-butoxycarbonyl-l-methYl-ethoxyimino)acetyl rhlor;rlr~ hydrochloride (1.84 g) under ice-cooling. /.ft~r being stirred for 1 hour, the reacticn mixture was added dropwise to ethyl ether (300 ml~, and the resulting precipitate was collected by filtration to give 78-[2-(5-amino-1,2,4-fh;~
3-yl) -2- (l-tert-butoxycarbonyl-l-methylethoxyimino) -acetamido ] -3- ( 3-amino-2 -methyl-l-pyrazolio ) methyl-3-cephem-4-carboxylate trihydrochloride ( syn isomer) (3.4 g).
IR (Nujol): 3300, 1780, 1720, 1650 cm 1 NMR (D2O, ~): 1.45 (9H, s), 1.57 (6H, s), 3.09 and 3.37 (2H, ~A3q, J=18Hz), 3.67 (3H, s), 35 4.98 ~nd 5.27 (2H, A~3q, J=15Hz), 5.21 (lH, d, - 32 - ~ ~i8320 J=5Hz), 5.86 (lH, d, J=5Hz), 5.92 (lH, d, J=3Hz), 7.85 (lH, d, J=3Hz) Example 6 The $ollowing compounds were obtained according to similar manners to those of Examples 1,3,4 and 5.
(1) 7~-~2-(5-Amino-1,2,4-~h;~fl;~ol-3-yl)-2-(1-tert-butoxycarbonyl-l-methylethoxyimino) acetamido] -3- ( 3-amino-2, 4 -dimethyl- l-pyrazolio ) methyl- 3-cephem-4-carboxylate trihydror hi ~rifl~ (syn isomer) IR (Nujol) : 3300, 1780, 1650 cm 1 NMR (D2O, ~) : 1.42 (9H, s), 1_47 (6H, s), 1.93 (3H, s), 3.32 (2H, broad s), 3.67 (3H, s), 5.18 (2H, broad s), 5.22 (lH, d, J=5Hz), 5.90 (lH, dd, J=8Hz, 5Hz), 7.90 (lH, s), 9.45 (lH, d, J=8Hz) (2) 73--[2--(5--Amino--1,2,4--~h;~fl;~7Ol--3--yl)--2--(l--carboxy-l-methylethoxyimino) acetamido] -3- (3-amino-2, 4-dimethyl-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol) : 3320, 3180, 1760, 1650, 1595 cm (3) 7,~-[2-(5-Amino-1,2,4-th;~fl;~7O1-3-yl)-2-(l-carboxy-l-methylethoxyimino) acetamido~ -3- (2-methyl-3-f ormamido- l-pyrazolio ) methyl-3- cephem-4 -carboxylate (syn isomer) IR (Nujol) : 3200-3300, 1760, 1580 cm NMR (DMSO-d6, ~): 1.46 (6H, s), 3.05-3.37 (2H, m), 3.91 (3H, s), 4.90-5.57 (2H, m), 5.06 (lH, d, J=5Hz), 5.71 (lH, dd, J=5, 8Hz), 6.91 (lH, d, J=3Hz), 8.02-8.27 (2H, br s), 8.34 (lH, d, J=3Hz), 8.56 (lH, s~, 9.46 (lH, d, J=8Hz) ~ 1 3388~0 (4) 7~-~2-(5--Amino-1,2,4-~h;~ 701-3-yl)-2-(l-carboxy-l-methylethoxyimino) acetamido] -3- [3-amino-2- (2-hydroxyethyl ) -l-pyrazolio ] methyl- 3-cephem-4 -carboxylate ( syn isomer) IR (Nujol): 3300, 1765, 1640 cm ND~R (D2O, ~): 1.58 (6H, s), 3.10 and 3.43 (2H, ABq, J=18Hz), 3.78-3.97 (2H, m), 4.26-4.46 (2H, m~, 5.15 (2H, br s), 5.26 (lX, d, J=5Hz), 5.87 (lH, d, J=5Hz), 5.97 (lH, d, J=3Hz), 7.89 (lH, d, J=3Hz) (5) 7~-[2-(5-Amino-1,2,4-~h;~ 7Ol-3-yl)-2-(l-carboxy-l-methylethoxyimino) acetamido] -3- (3-acetamido-2-methyl-l-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol): 3300, 1775, 1670 cm 1 N~R (D2O, ~) : 1.56 (6H, s), 2.31 (3H, s), 3.20 and 3.50 (2H, A;3q, J=18Hz), 3.93 (3H, s), 5.23 and 5.47 (2H, ABq, J=15Hz), 5.26 (lH, d, J=5Hz), 5.88 (lH, d, J=5Hz), 6.88 (lH, d, J=3Hz), 8.19 (lH, d, J=3Hz) (6) 7~-[2-(5-Amino-1,2,4-~h;~ 7ol-3-yl)-2-(l-carboxy-l-methylethoxyimino) acetamido] -3- (2-methyl-3-ureido-1-pyrazolio)methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol): 3300 (broad s), 1770, 1680, 1570 cm 1 Example 7 Trifluoroacetic acid ( 7 ml) was added dropwise to a suspension of 7~- [2- (5-amino-1,2 ,4-th; R~; ~7Ql-3-yl) -2 - ( l-tert-butoxycarbonyl-l-methylethoxyimino ) acetamido ] -3- ( 3-amino-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate trihydrochloride (3.3 g) and anisole (3.5 ml) in methylene chloride (10 ml) at ambient temperature.

After being stirred at the same temperature ~or 4 hours, the mixture was poured into diisopropyl ether (300 ml), and the resultLng precipitate was collected by filtration. The precipltate was dis601ved in water (100 ml), and the solution was adjusted to pX 2 with 5 aqueous solution of sodium hi~!~rh~ ti . The aqueous solution was subjected to colul[n chromatography on ma~:luL~-uu~ non-ioniC adsorption resin "Diaion HP-20" .
The desired product was eluted with 5% aqueous isopropyl alcohol solution and lyo~hi 1; 7ed to give 7~- [2- (5-amino-1,2,4-~h;Arl;~7ol-3-yl)-2-(l-carboxy-l-methylethoxy-imino) acetamido] - 3 - (3-amino-2-methyl-1-pyrazolio) -methyl-3-cephem-4-carboxylate (syn isomer) (515 mg).
IR (Nujol) : 3325, 1770, 1650, 1630, 1590 cm 1 NMR (D2O, ô): 1.52 (6H, s), 3.19 and 3.37 (2H, ABq, J=18Hz), 3.66 (3H, s), 4.97 and 5.25 (2H, A~3q, J=15Hz), 5.20 (lH, d, J=5Hz), 5.84 (lH, d, J=5Hz), 5.91 (lH,' d, J=3Hz), 7.82 (lH, d, J=3Hz) Example 8 The following compounds were obtained according to a similar manner to that of Example 7.
(1) 7~-[2-(5-Amino-1,2,4-th;z~ 7Q1-3-yl)-2-(l-carboxy-1-methylethoxyimino ) acetamido ] -3- ( 3-amino-2, 4-dimethyl-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol): 3320, 3180, 1760, 1650, 1595 cm 1 NMR (D2O, ~): 1.60 (6H, s), 1.96 (3H, s), 3.10 and 3.37 (2H, A}3q, J=18Hz), 3.68 (3H, s), 4.92 and 5.23 (2H, A~3q, J=15Hz), 5.22 (lH, d, J=5Hz), 5.86 (lH, d, J=5Hz), 7.68 (lH, s) 35 (2) 78--[2--(5--ATnino--1,2,4--~h;~ 7ol--3--yl)--2-.
carboxymethoxy; m; nnAr-.etamido] -3- (4-formamido-2-methyl-l-pyrazolio) methyl-3-cephem-4-carboxylate ~syn isomer) IR (Nujol): 3300, 1765, 1665, 1605 cm 1 (3) 7~-- [2--(5--Amino--1,2 ,4--~h; A~; A7ol--3--yl) --2--carboxymethoxyiminoacetamido] -3- (3-fnrr-m; cln-2-methyl-l-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nu~ol): 3300, 1760, 1660, 1580 cm 1 (4 ) 7 3-- [2-- (5--Amino--1, 2, 4--th; A~l; A 701--3--yl) --2--carboxymethoxy; m; nnA~ ~tamido] -3- ~3-formamido-2, 4-dimethyl-1-pyrazolio) met~yl-3-cephem-4-carboxylate (syn isomer) R (Nujol): 3300, 1765, 1660, 1600 cm 1 5) 7~-- [2-- (5--Amino--1,2, 4--~h; A~; A70l--3--yl) --2--CaLIJUlLy ~hoxy;m;nnA~etamldo]-3-(3-amino-2-methyl-l-pyrazolio) methyl-3 c~:ph~l,. 4-earboxylate (syn isomer) IR (Nu~ol): 3300, 1760, 1660, 1590 cm 1 ~6 ) 7~-- [2-- (5--Amino--1, 2, 4--th; A;l; A7r>l --3--yl) --2--eal~ol~y - thoxyiminoacetamido ] - 3- ( 3-amino-2, 4 -dimethyl-l-pyrazolio) methyl-3-eephem-4-earboxylate ( syn i somer) IR (Nujol): 3350, 1770, 1660, 1600 em 1 (7) 7~--[2--(5--Amino--1,2,4--th;a-l;A7ol--3--yl)--2--(l--earboxy-l-methylethoxyimino) acetamido] -3- (2-methyl-3-forr-m; r1o-l-pyrazolio) methyl-3-eephem-4-carboxylate (syn isomer) IR (Nujol) : 3200-3300,'1760, 1580 em - 36 - ~ 3 3 8 8 2 0 (8) 7~--[2--(5-Amino--1,2,4--~hi~ 701--3--yl)--2--(1--carboxy-l-methylethoxyimino) acetamido] -3- [3-amino-2 - ( 2 -I1YdLO~Y e: Ll~y 1 ) -l-pyrazolio ] methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol): 3300, 1765, 1640 cm 1 (9) 73-[2-(5-Amino-1,2,4-~h;~ 701-3-yl)-2-(l-carboxy-l-methylethoxyimino) acetamido] -3- (3-acetamido-2-methyl-l-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol): 3300, 1775, 1670 cm 1 (10) 7~-[2-(5-Amino-1,2,4-~h;~fl;~701-3-yl)-2-(l-carboxy-l-methylethoxyimino) acetamido] -3- (2-methyl-3-ureido-l-pyrazolio) methyl-3-cephem-4-carboxylate (~yn isomer) IR (Nu~ol): 3300 (broad s), 1770, 1680, 1570 cm 1 (to be continued to the next page) ~37 Preparation 10 To a solution of r~lonnn;trile (300 g) and acetic acid (54. 6 g) in acetonitrile (2.1 Q) was added sodium nitrite (313 g) at 60-65C. After being stirred at the same temperature for an hour, ethyl 2-bromo-2-methylpropanoate (797 g) was added to the mixture at 65C. After being refluxed for five hours, the mixture was cooled to the ambient temperature and allowed to stand overnight. The resulting mixture was poured into a mixture of water (4.2 Q) and diisopropyl ether (4.2 Q).
The separated organic layer was washed with water and brine, dried over magnesium sulfate and evaporated under reduced pressure to s~ive 2- (l-ethoxycarbonyl-l-methylethoxyimino) pror~n~Ai n; trile (860 g) .
IR (Neat): 3000, 2250, 1750 cm 1 NMR (CDC13, ô): 1.30 (t, J=7Hz, 3H), 1.69 (s, 6H), 4.25 (q, J=7Hz, 2H) Preparation 11 To a solution of ammonium acetate (1258 g) in methanol (8 . 54 Q) were added 2- (l-ethoxycarbonyl-l-methylethoxyimino)prop~no~lin;trile (854 g) and 28~ ammonia water (568 ml). After the mixture was stirred at 20C
for 15 hours, methanol was evaporated under reduced pre-ssure. The residue was dissolved in a mixture of water (8 Q) and tetrahydrofuran (8 Q). After the aqueous layer was saturated with sodium nhl nr; ~ the organic layer was separated and dried over magnesium s~llfate. Acetic acid (368 g) was added to the solution, t:hen the mixture was evaporated under reduced pressure, and the residue was triturated with diisopropyl ether ( 8 Q) to give 2-cyano-2- (l-ethoxycarbonyl-l-methylethoxyimino) acet~ri ~1; nf-acetate (689.1 g).
IR (Nujol): 1750, 1670 cnl 35 N~lR (CDC13, ô): 1.26 (t, J=7Hz, 3H), 1.67 (s, 6H), 2.00 (s, 3H), 4.23 (q, J=7Hz, 2H), 8.73 (broad s, 2H) Preparation 12 To a solution of 2-cyano-2- (l-ethoxycarbonyl-l-methylethoxyimino) ace~m; ~; n~ acetate (685 g) in methanol (6. 85 Q) was added triethylamine (557 g) at -13 ~ -15C
over a 20-minute period and then bromine (344 g) was added thereto at the same temperature over the same period.
The solution was stirred at -13 ~-15C for lS minutes, then N,N-dimethylfor~=~ e (685 ml) was added to the solution at the same temperature, and a solution of potassium thiocyanate (209 5) in methanol (2 . 09 ~) was added thereto at -13 ~-15C over a 30-minute period.
After being stirred at -5 ~ ODC for an hour, the solution was poured into a cold water (5 ~10C) and stirred for an hour under ice-cooling. The resulting precipitates were collected by filtration, washed with~cold water and dried to give 2-(5-amino-1,2,4-~h;~;A7~1-3-yl)-2-(l-ethoxycarl~onyl-l-methylethoxyimino) acetonitrile (390 g) .
IR (Nujol): 3470, 3280, 3140, 1730, 1625, 1540 cm NMR (CDC13, ~) : 1.25 ~t, J=7Hz, 3E~), 1.67 (s, 6H), 4.23 (q, J=7Hz, 2H), 7.86 (broad s 2~I) . 5 Preparation 13 A mixture of 2-(5-amino-1,2,4--~h;~ 701-3-yl)-2-(l-ethoxycarbonyl-l-methylethoxyimino) acetonitrile (8.4 g) in 2N sodium hydroxide was stirred at 60-65C
for 6 hours, adjusted to pH 4.2 with 6N hydrochloric acid under ice-cooling and washed with ethyl acetate (100 ml).
Tetrahydrofuran (120 ml) was added to the aqueous solution, and the solution was adjusted to pH 1. 0 with 6N hydrochloric acid under ice-cooling and saturated with sodium chloride.
The separated organic layer was evaporated under reduced . ~ 39 1 338820 pressure. The residue was dissolved in a saturated a~ueous solution (90 ml) of sodium hic~rhQn~te and activated carbon (200 mg) was added thereto.
After insoluble material and activated carbon were removed by filtration, the solution was adjusted to pH 1. 0 with 6N hydrochloric acid under ice-cooling. The resulting precipitates were collected by filtration, washed with cold water and dried to give a crude produci; (6 . g6 g) .
The crude product was recrystallized from propyl alcohol (111 ml~ to give (Z)-2-(S-amino-1,2,4-~h;~ 7Ql-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetic acid (4 . 70 g) .
IR ( Nu~ol): 3~30, 3280, 3130, 1740, 1680, 1635, 1540 cm 1 15 NMR (DMSO-d6, ~) : 1.46 (s, 6H), 8.23 (broad s, 2H) Preparation 14 A solution of (Z)-2-(5-amino-1,2,4-~ 7ol-3-yl) -2- (l-tert-L,~lt~"ycdrbonyl-l-methylethoxyimino) acetic acid (17 . 8 g) and anisole (18 ml) in trifluoroacetic acid (36 ml) was stirred for 4 . 0 hours at room temperature.
The reaction mixture was concentrated under reduced pressure and an aqueous solution of sodium bi~ Arhor~te was added thereto to adjust to pH 6. 0 . The aqueous solution was washed with ethyl acetate, ad~usted to pH 3. 8 with 6N
hydrochloric acid and washed with ethyl acetate. The aqueous layer was separated and 6N hydrochloric acid was added thereto to adjust to pH 1. 0 . After the mixture was stirred for 10 minutes, the resultant precipitate was collected by filtration to give (Z)-2-(5-amino-1,2,4-~h; ~ 701-3-yl) -2- (1-carboxy-1-methylethoxyimino) acetic acid (13 . 0 g) .
Preparation 15 33 ~o ~ ~olut1on o~ (3~-2-lS-am n~-1,2,4-thif~l;A7ol-3--~o -yl)-2-(1-carboxy-1-methylethoxyimino)acetic acid (2.74 g) in N,N-dimethylacetamide (45 ml~ were added methanesulfonyl chloride (1.15 g) and potassium bicarbonate (1.20 g) under cooling in an ice bath.
The mixture was stirred for 2.5 hours at 5C and poured into a cold mixture of water (200 ml), ethyl acetate (200 ml) and lN hydrochloric acid (6 ml). The mixture was stirred for 5 minutes under cooling in an ice bath. The organic layer was separated, washed with cold water (200mlx2) and with a cold saturated aqueous solution of sodium chloride, dried over magnesium sulfate and evaporated.
Toluene was added to the residue. To the mixture was added methylene chloride and the mi~ ure was cooled for 10 minutes in an ice bath. The crystals were collected by filtration, washed with methylene chloride and air-dried to give methanesulfonyl (Z)-2-(5-amino-1,2,4-~h;A~q;A7ol-3-yl) -2- (l-carboxy-l-methylethoxyimino) acetate (1.1 g) .
Example 9 A mixture of 7~-amino-3- (3-amino-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate trihydrochloride (0 . 42 g), N- (trimethylsilyl) acetamide (2 . 5 g) and tetrahydrofuran was stirred for 1. 0 hour at room temperature.
~ethanesulfonyl (Z)-2-(5-amino-1,2,4-~i~;A~;A7ol-3-yl) -2- (l-carboxy-l-methylethoxyimino) acetate (350 mg) was added thereto at room temperature and stirred for 2. 0 hours at the same temperature. The mixture was poured into diisopropyl ether (50 ml), and the precipitates were collected by filtration, dissolved in water (50 ml), adjusted to pH 2 . 0 with an aqueous solution of sodium bicarbonate and washed with ethyl acetate, and ethyl acetate in the aqueous layer was evaporated. The aqueous layer was sub j ected to column chromatography on ma.;L~,~oLuus non-ionic adsorption resin "Diaion HP-20" and the elution was carried out with 30% aqueous methyl ~ 41 ~ 1 3 3 8 8 20 ~lcohol solution. Methyl alcohol in the ~ractions con-taining the ob~ ect compound was evaporated and the residue was lyorhi l; 7e-1 to give 78- [2- (5-amino-1,2,4-~h; ~ 70l-3-yl) -2- (l-carboxy-l-methylethoxyimino) -acetamido]-3- (3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate (syn isomer) (330 mg).
IR (Nujol): 3325, 1770, 1650, 1630, 1590 cm NMR (D2O, ~): 1.52 (6H, s), 3.19 and 3.37 (2H, ABq, J=18Hz), 3.66 (3H, s), 4.97 and ; 5.25 (2H, ABq, J~15Hz), 5.20 (lH, d, J--5Hz), 5.84 (lH, d, J=5Hz), 5.91 (lH, d, J=3Hz), 7 . 82 (lH, d, J=3Hz) .
Prepar~tion 16 7 ~ -Amino- 3- ( 3-amino-2-methyl- l-pyrazo lio) methy 1-3-cephem-4-carboxylate trihydrochloride ( 5 kg) was dissolved in water (16 ~) . This solution. was subjected to column chromatography on macroporous non-ionic adsorption reslin "Diaion HP-20".
The desired product was eluted with water. To the eluate (30 Q) was added acetone (160 ~) and the mixture was stirred at room temperature for 2 hours.
The resulting precipitate was collected by filtration to give 7~-amino-3- (3-amino-2-methyl.-1-pyrazolio)methyl-3-cephem-4-carboxylate hydrochloride dihydrate as crvstals -(1. 802 kg), IR (Nujol): 3540 , 3350 , 3150 , 1775 , 1635 , 1585 cm 1 NMR (DMSO--d6, ~): 3.66 (3H, s), 4.83 (lH, d, J=5Hz), 5,03 (lH, d~ J=5Hz), 5.18 and 5,30 (2H, ABq, J=15Hz), 5.85 (lH, d, J=3Hz), 7.44 (2H, broad s) 8.08 (lH, d, J=3Hz) Example 10 To N,N-aimethylforr-mifl~ (231.6 ml~ was adaed 7B- [2- ( 5-amino-1, 2, 4-th; ~fl i ~ 701-3-yl) -2- ( 1-carboxy-1-methylethoxyimino) acetamido] -3- (3-amino-2-methyl-1-pyrazolio)methyl-3-cephem-4-carboxylate (syn isomer) (38.6 g) at ambient temperature. The mixture was stirred at the same temperature for 2 hours, and the resultant precipitate was collected by filtration to give bis(N,N-dimethylform~mi~e) solvate (47.3 g) of 7B-[2-(5-amino-1,2,4-~h;~fl;~ol-3-yl)-2-(1-carboxy-1-methylethoxyimino) acetamido] -3- (3-amino-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) .
IR (Nujol): 3280, 3130, 1775, 1670, 1580 cm 1 NMR (D2O+NaHCO3, ~): 1.53 (6H, s), 2.86 (6H, s).
3.01 (6H, s), 3.10 and 3.36 (2H, ABq, J=18Hz), 3.66 (3H, s), 4.96 and 5.23 (2H, ABq, J=15Hz), 5.22 (lH, d, J=5Hz), 5.85 (lH, d, J=5Hz), 5.92 (lH, a, J=3Hz), 7.83 (lH, d, J=3Hz), 7.91 (2H, s) Example 11 To a 601ution of 7B-[2-(5-amino-l~2~4-th;~fl;~ol-3-yl) 2- (l-carboxy-1-methylethoxyimino) acetamido] -3- (3-amino-2-methyl-l-pyrazolio) methyl-3-cephem-4-carboxylate (syn 25 - isomer) (0 . 5 g) in an aqueous sulfuric acid (2M, 1. 0 ml) was added ethanol. After the solution was stirred for 1. 0 hour, the crystals were collected by filtration, washed with a solution of water and ethanol ( 1: 5 ), then washed with ethanol and dried over phosphorus pentoxide to give sulfuric acid salt (480 mg) of 7B-[2-(5-amino-1,2,4-~h;~fl;~ol-3-yl)-2- (l-carboxy-1-methylethoxyimino) acetamido]-3- (3-amino-2-methyl-l-pyrazolio)methyl-3-cephem-4-carboxylate (syn isomer).

l 3388~0 mp: 194-197C
IR (Nu~ol): 3320, 3200, 3060, 1770, 1720, 1655, 1590, 1545 cm~l NMR (D2O, ~): 1.50 (6H, s), 3.33, 3.13 (2H, ABq, J=18Hz), 3. 65 (3H, s), 5 . 20 (lH, d, J=5~z), 5.22, 4.98 (2H, ABq, J=14Hz), 5.83 (lH, d, J=5Hz), 5.92 (lH, d, J=3Hz), 7. 80 (lH, d, J=3Hz) (to be continued to the next page) ~ 338820 Preparation 17 To a solution of sulfuryl ~hlor;~l~ (105.15 g) in methylene chloride (1500 ml) was added a solution of triphenyl phosphite (279 g) in methylene chloride (300 ml) at -20 C and the mixture was stirred for 30 minutes at -20 ~ -30C. To the mixture was added benzhydryl 78- (2-hydroxybenzyl; 1PnP~m;n- ) -3-1-y-1Lo~y thyl-3-cephem-4-carboxylate (300 g) at -20 ~ -30C. The mixture was stirred for 30 minutes at -20 ~ -30 C and poured into a mixture of ethyl acetate (7.5 Q) and 6.5%
a~aueous potassium carbonate (2.4 Q). The organic layer was separated, washed with brine (300 ml) and concentrated in vacuo to 500 ml at 30 C. To the concentrated solution were added N,N-dimethyl ~orr-m; de (300 ml), 5-formamido-1-methylpyrazole (187.5 g) and potassium iodide (119.4 g) and the mixture was stirred for 22 hours at 31 ~ 34C. The reaction mixture was diluted with acetone (600 ml) and the dilute solution was added dropwise into the mixture of isopropyl alcohol (6.0 Q) and diisopropyl ether (6.0 Q)at 20 ~ 25C.
The resulting precipitate was collected by filtration and dried in vacuo to give benzhydryl 73- (2-hydroxy-benzyli~PnP~m; nn) -3- ( 3-formamido-2-methyl-1-pyrazolio ) -methyl-3-cephem-4-carboxylate iodide (514. 8 g) .
N~R (DMSO-d6, ~) : 3.60 (2H, br s), 3.75 (3H, s), 5.50 (2H, br s), 5.33 (lH, d, J=6Hz), 5.90 (lH, d, J=6Hz), 6.60-7.60 (15H, m), 8.33-8.60 (3E, m), 8.90 (lH, s), 12.00 (lH, m) IR (Nujol): 1790, 1725, 1630, 1590, 1230, 1110 cm 1 Preparation 18 To a solution of benzhydryl 16-(2-hydroxybenzylidene-amino) -3- ( 3-formamido-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate iodide (170 g) in methylene chloride ~ f 338820 and formic acid was added 35~ hydrochloric acid (24.65 g).
The mixture was stirred for 3 hours at 25 ~ 30C and added dropwise $nto the mixture of acetone (1700 ml) and ethyl acetate (3400 ml). The resulting precipitate was collected by filtration and dried in vacuo to give 73-amino-3- ( 3-fr r~ ; flr--2-methyl-l-pyrazolio) methyl-3-cephem-4-carboxylate hydrochloride ( 86 . 7 g) .
IR (Nujol): 1800, 1720, 1650, 1590 cm 1 NMR (DMSO-d6, ~): 3.55 (2H, br s), 4.03 (3H, s), 5.30 (2H, br s), 5 . 33 (2H, br s), 6.80-7.60 (2H, m), 8.60 (1~, br s) Preparation l9 To a solution of benzhydryl 7~.- (2-hydroxybenzylidene-amino ) -3- ( 3-fr rr--~; flr--2-methyl -l-pyrazolio ) methyl-3-cephem-4-carboxylate iodide (510 g) in methylene chloride (1275 ml) and formic acid (1275 ml) was added 3596 hydrochloric acid (145 g) at 20 ~ 25C. The mixture was stirred for l hour at 20 ~ 25C and added dropwise into the mixture of acetone (1020 ml) and ethyl acetate (2040 ml) at the same temperature. The resulting precipitate was collected by filtration and dried in vacuo to give the mixture (255.9 g) of 78-amino-3- ( 3-formamido-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate hydrochloride and 7~-amino-3-(3-amino-2-methyl-l-pyrazolio) methyl-3-cephem-4-carboxylate hydrochloride . The mixture (255 . 9 g) was dissolved in methanol (1023.6 ml). To the solution was added 35%
hydrochloric acid (66.6 g) and the mixture was stirred for 1 hour at 28 ~ 30C. The insoluble material was filtered off and washed with methanol (512 ml). The combined filtrates and washings were added dropwise into the mixture of acetone (2560 ml) and ethyl acetate (5120 ml) at 20 ~ 25C. The resulting precipitate was collected by filtration and dried in vacuo to give ~ 46 ~ 1 338820 7~ -amino- 3- ( 3-amino-2 -me thy l-l-pyrazolio ) methyl- 3-cephem-4-carboxylate hydrochloride.
IR (Nujol): 1800, 1720, 1650, 1600, 1230 cm N~5R (DMSO-d6, ô) : 3.30 and 3.55 (2E, Asq, J=18Hz), 3.70 (3H, s~, 5.25 (2H, br s), 5.33 (2H, br s), 5.93 (lH, d, J=3Hz), 8.25 ~lH, d, J=3Hz) (to be continued to the next page) .

Preparation 20 A solution of sodium cyanate (52 g) in water (400 ml) was added dropwise to a solution of l-methyl-5-aminopyrazole (19.4 g) in acetic acid (96 ml) and water (192 ml), and the mixture was stirred at ambient temperature for 7 hours. The reaction mixture was poured into a mixture of water (400 ml) and ethyl acetate (400 ml). The organic layer was separated, washed with saturated aqueous sodium chloride, dried over magnesium sulfate and l:::VCl~L.ll~l to give l-methyl-5-ureidopyrazole (14 . 6 g) .
IR (Nujol): 3300 (broad s), 1735 , 1600 (broad s) cm 1 NMR (DMSO--d6, ~): 3.63, 3.67 (3H, d, J=2Hz), 6.10, 6.22 (lH, dd, J=2H~), 7.28, 7.35 (lH, d, J=2~ z ) .
Preparation 2 1 A mixture of acetic anhydride (11.13 ml) and formic acid (5 . 93 ml) was stirred at ambient temperature for 30 minutes . To this solution was added 5-amino-1- (2-hydroxyethyl)pyrazole (5 g) under ice-coolinq, and the mixture was stirred at 30-40C for 1 hour. The reaction mixture was poured into a mixture of water, tetrahydro-furan and ethyl acetate and adjuc~:ed to pH 6 with a~aueous sodium bicArhnnAt~. The organic layer was separated, and the aqueous layer was extracted with a mixture of tetrahydrofuran and ethyl acetate for three times. The organic layers were combined, dried over magnesium sulfate and evaporated in vacuo to give 5-formamido-1- (2-formyloxyethyl) pyrazole (5 .18 g) .
IR (Nujol): 3180, 1705, 1660 cm N~R (DMSO-d6, ~) : 4.21-4.61 (4H, m), 6.11 and 6.34 (lH, each d, J=3Hz), 7.47 (lH, d, J=3Hz), 8.00 (lH, s), 8.33 '(lH, s) .

Preparation 22 The following compounds were obtained according to a similar manner to that of Preparation 3.
(1) Benzhydryl 7~-tert-l,uLv,~y-:arbonylamino-3- (3-acetamido-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate iodide IR (Nujol) : 1780, 1710 cm NMR (DMSO--d6, ~) : 1.44 (9E~, s), 2.25 (3H, ~), 3.43 (2H, l~r s), 3.74 (3H, s), 5.16 (lH, d, J=5Hz), 5.38 (2H, br s), 5.63 (lH, dd, J=8, 5Hz), 6.93 (lH, d, J=3Hz), 6.94 (lH, s), 7.15--7.55 (lOH, m), 7.97 (lH, d, J=8Hz), 8.25 (lH, d, J=3Hz), 11.13 (lH, s) (2) Benzhydryl 7~-tert-butoxycarbonylamino-3- [3-formamido-2- (2-formyloxyethyl) -l-pyrazolio]methyl-3-cephem-4-carboxylate iodide IR (Nujol) : 1780, 1720 cm 20NMR (DMSO-d6, ô): 1.49 (9H, s), 3.43 (2H, br g), 4.14-4.38 (2H, m), 4.52-4.73 (2H, m), 5.15 (lH, d, J=5Hz), 5.40 (2H, br s), 5.67 (lH, dd, J=5, 8Hz), 6.88 (lH, s), 7.02 (lH, d, J=3Hz), 7.18-7.52 (lOH, m), 7.94 (lH, d, J=8Hz), 7.99 (lH, s), 8.27 (lH, d, J=3Hz), 8.51 (lH, br s) Preparation 2 3 The following compounds were obtained according to a similar manner to that of Preparation 5.
(1) Di (trifluoroacetic acid) salt of 7~-amino-3- [3-formamido-2- ( 2-formyloxyethyl) -l-pyrazolio] methyl-3-cephem-4-carboxylate 35 IR (Nuiol) : 1780, 1715, 1660 cm ~ ~ 338820 NMR (DMSO-d6, ~): 3.53 (2H, br s), 4.28-4.56 (2H, m), 4.78-4.99 (2H, m), 5.29 (2H, br s), 5.53 (2H, br s), 7.14 (lH, d, J=3Hz), 8.22 (lH, s), 8.46 (lH, d, J=3Hz), 8.63 (lH, s) (2) Di(trifluoroacetic acid) salt of 7~-amino-3-(3-acetamido-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxyl ate IR (Nujol) : 1780, 1670 cm NMR (DMSO-d6 , ~) : 2.24 (3H, s), 3.47 (2H, br s), 3.93 (3H, s), 5.22 (2H, s), 5.50 (2H, br s), 6.98 (lH, d, J=3Hz), 8.35 (lH, d, J=3Hz) Preparation 24 The following compound was obtained by reacting di(trifluoroaoetic acid) salt of 73-amino-3-[3-f 2- (2-formyloxyethyl) -1-pyrazolio]methyl-3-cephem-4-carboxyla~e according to a similar manner to that of Preparation 7.
7~-Amino-3-[3-amino-2-(2-hydroxyethyl)-1-pyrazolio]-methyl-3-cephem-4-carboxylate trihydrochloride IR (Nujol) : 3250, 1770, 1700, 1625 cm NMR (DMSO-d6, ~): 3.43 (2H, br 5), 3.52-3.88 (2H, m), 4.18-4.48 (2H, m), 5.28 (2H, br s), 5.37 (2H, br s), 5.97 (lH, d, J=3Hz), 8.18 (lH, d, J=3Hz) Preparation 25 To a solution of ammonium acetate (73.7 g) in methanol (250 ml) were added 2- (l-ethoxycarbonyl-l-methylethoxyimino)propanedinitrile (50.0 g) and 28~
ammonia water (33. 3 ml) at room temperature. After stirring for 15 hours at 20C, potassium carbonate (33.0 g) was added to the solution,' and the solution was 35 evaporated under reduced pressure. A mixture of water (130 ml) and methylene chloride (110 ml) was added into the residue . The mixture was adjusted to pH 8. 0-8. 2 and extracted with methylene ~hl nr; ~P twice . Phthalic acid (35.7 g) was added to the extract at 20-30C, and then diisopropyl ether (200 ml~ was added thereto.
~f ter stirring ~or 2 hours at the same temperature, the resulting precipitate was collected by filtration, washed with diisopropyl ether (40 ml) and dried under reduced pressure to give phthalic acid salt (61.4 g) of 2-cyano-2- ( l-ethoxycarbonyl-l-methylethoxyimino) acetamidine .
mp: 156-158C
IR (Nujol): 3380, 1730, 1700 cm NMR (DMSO-d6, ~): 1.20 (3H, t, J=7Hz), 1.66 (6H, 8), 4.17 (2H, q, J=7Hz), 7.4-7.7 (2H, m), 7 . 9 -8 . 2 ( 2H, m), 12 . 0 ( 5H, broad s ) Preparation 26 To a suspension o~ phthalic acid salt (10.0 g) of 2-cyano-2- (l-ethoxycarbonyl-l-methylethoxyimino) -acet~m; ,1; n~ in methanol (60 ml) were added dropwise triethylamine (9.01 g) and bromine (3.67 g) at -15~-10C.
After stirring for 15 minutes at the same temperature, N,N-dimethylformamide (6.0 ml) and a solution of potassium thiocyanate (2.23 g) in methanol (22.3 ml) were added dropwise thereto at -15~-10C. After stirring for one hour at -5~0C, the solution was poured into cold water ( 300 ml) and the mixture was stirred for one hour under ice-cooling. The resulting precipitate was collected by filtration, washed with cold water (60 ml) and dried under reduced pressure to give 2-(5-amino-1,2, 4-th; ~rl; ~ol-3-yl) -2- (l-ethoxycarbonyl-l-methylethoxyimino) -acetonitrile ( 3 . 82 g) .
IR (Nujol): 3470, 3280, 3140, 1730, 1625, 1540 cm ~ 1 338820 Example 12 l-Methyl-5-ureidopyrazole (1.4 g) was added to a solution of 7~- [2- (5-amino-1,2 ,4-~hi ~ 701--3--yl)--2- (1-carboxy-l-methylethoxyimino) acetamido]-3-chloromethyl-3-cephem-4-carboxylic acid trifluoroacetate (syn isomer) (1.24 g) in N,N-dimethylf~ (13 ml) and the mixture was stirred at ambient temperature for 4 hours. The reaction mixture was poured into ethyl acetate ~100 ml).
The precipitates were collected by filtration and successively washed with ethyl acetate and diisopropyl ether, and the solid was dissolved in water ~30 ml) and adjusted to pH 2. 0 with 10% hydrochloric acid. The solution was sub j ected to column chlomatography on ma,:L~poL~,us non-ionic adsorption resin "Diaion HP-20"
and eluted with 30% aqueous solution of methyl alcohol.
The fractions containing the object compound were collected, concentrated in vacuo and lyophilized to give 7~[2-~5-amino-1,2,4-~h;~l;a7- 1-3-yl)-2-~l-carboxy-l-methylethoxyimino ) acetamido ] -3- ~ 2 -methyl- 3-ureido-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) (0.12 g).
IR (Nujol) : 3300 (broad s), 1770, 1680, 1570 cm 1 N~R (D20, ô) : 1.53 (6H, s), 3.14, 3.44 (2H, ABq, J=18Hz), 3.70 (3H, s), 5.22 (lH, d, J=5Hz), 5.27 (2H, broad s), 5.85 (lH, d, J=5Hz), 6.75 (lH, d, J=3Hz), 8.10 (lH, d, J=3Hz) Example 13 The following compounds were obtained according to a similar manner to that of Example 12.
(1) 7~-[2-(5-Amino-l~2~4-~h;~ 7ol-3-yl)-2-carboxy-methoxyiminoacetamido]-3- (4-formamido-2-methyl-1-pyrazolio) methyl-3-cephein-4-carboxylate (syn isomer) 35 IR (Nujol): 3300, 1765, 1665, 1605 cm 1 3388~0 (2) 7~~ [2--(5--Amino--1, 2 ,4--th; A~l; A7' l--3--yl) --2--carboxymethoxy; m; nr~cetamido] -3- (3-forr-m; ~n-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate ( syn isomer) IR (Nu~ol): 3300, 1760, 1660, 1580 cm (3) 7~--[2--(5--Amino--1,2,4--~h;A~ ol--3--yl)--2--carboxymethoxy;m;nr-~cetamido] -3- (3-formamido-2, 4-dimethyl-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol): 3300, 1765, 1660, 1600 cm (4) 7~--[2--(5--Amino--1,2,4-th;A~ 7ol--3--yl)--2--carboxy-methoxy;m; nr~ etamido] -3- (3-amino-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol) : 3300, 1760, 1660, 1590 cm (5) 7~- [2- (5-Amino-1, 2 ,4-~h; ~ 701-3-yl) -2-carboxy-methoxyiminoacetamido ] - 3- ( 3-amino-2, 4-dime thyl-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol): 3350, 1770, 1660, 1600 cm 1 (6) 7~-[2-(5-Amino-1,2,4-~h;Afl;~701-3-yl)-2-(l-tert-butoxycarbonyl -l-methylethoxyimino ) acetamido ] -3-(3-amino-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate trihydrochloride (syn isomer) IR (Nujol): 3300, 1780, 1720, 1650 cm 1 ( 7) 7~- [2- ( 5-Amino-1, 2, 4-~h; Arq; A 7:ol-3-yl ) -2- (l-tert-butoxycarbonyl-l-methylethoxyimino) acetamido] -3- ( 3-amino-2, 4-dimethyl-1-pyrazolio ) methyl-3-cephem-4-carboxylate trihydrochloride (syn isomer) IR (Nujol): 3300, 1780, 1650 cm 1 ( 8 ) 7 8 - [ 2 - ( 5 -Am~ ino~ 1, 2, 4 -thi adi a z ol - 3 -yl ) - 2 - ( 1-carboxy-l-methylethoxyimino) acetamido] -3- (3-amino-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nu~ol): 3325, 1770, 1650, 1630, 1590 cm (9) 7~--[2--(5--Amino--1,2,4--th;;~ q7ol--3--yl)--2--(l--. . carboxy-l-methylethoxyimino) acetamido] -3- (3-amino-2, 4 -dimethyl-l-pyrazolio ) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol): 3320, 3180, 1760, 1650, 1595 cm 1 (10) 7~--[2--(5--Amino--1,2,4--th;~ 701--3--yl)--2--(1--carboxy-l-methylethoxyimino) acetamido] -3- (2-methyl-3-formamido-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol): 320~-3300, 1760, 1580 cm (11) 7~- [2- (5-Amino-1,2 ,4-~; ;'fl; ~ol-3-yl) -2- (1-carboxy-l-methylethoxyimino) acetamido] -3- [3-2 0 amino-2 - ( 2-hydroxyethyl ) - l-pyrazolio ] methyl- 3-cephem~4-carboxylate (syn isomer) IR (Nujol) : 3300, 1765, 1640 cm (12) 7~-[2-(5-Amino-1,2,4-~ zol-3-yl)-2-(1-carboxy-l-methylethoxyimino) acetamido]-3- (3-acetamido-2-methyl-1-pyrazolio) methyl-3-cephem-4-carboxylate (syn isomer) IR (Nujol) : 3300, 1775, 1670 cm 1

Claims (2)

1. A compound of the formula:
wherein R3 is lower alkyl, hydroxy (lower) alkyl or a protected hydroxy (lower) alkyl, R4 is amino or a protected amino group, and R5 is hydrogen or lower alkyl, and a salt thereof.

2. A process for preparing a compound of the formula:
wherein R3 is lower alkyl, hydroxy (lower) alkyl or a protected hydroxy(lower)alkyl, R4 is amino or a protected amino group, and R5 is hydrogen or lower alkyl, or a salt thereof, which comprises (1) subjecting a compound of the formula:
wherein R4 and R5 are each as defined above, R3 is hydroxy (lower) alkyl or a protected hydroxy (lower) alkyl, R6 is a protected amino group, R7 is a protected carboxy group, and X? is an anion, or a salt thereof to elimination reaction of the amino protective group in R6 and the carboxy protective group in R7 to give a compound of the formula :
wherein R3, R4 and R5 are each as defined above, or a salt thereof, or (2) subjecting a compound of the formula :

wherein R3a and R5 are each as defined above, and R4a is a protected amino group, or a salt thereof to elimination reaction of the amino protective group to give a compound of the formula:
wherein R3 and R5 are each as defined above, or a salt thereof, or (3) subjecting a compound of the formula:
wherein R4, R5, R7 and X(-)are each as defined above, R3b is lower alkyl, and R6a is hydroxybenzylideneamino, or a salt thereof to elimination reaction of the amino protective group in R6a and the carboxy protective group in R7 to give a compound of the formula:

wherein R3b, R4 and R5 are each as defined above, or a salt thereof, or (4) subjecting a compound of the formula :
wherein R5a is hydrogen or methyl, or a salt thereof to deformylation to give a compound of the formula:
wherein R5a is as defined above, or a salt thereof.