CA1231940A - Cephalosporin compounds and preparation thereof - Google Patents

Abstract

ABSTRACT
A cephalosporin compound of the formula:

(I) wherein R1 is hydrogen or lower alkyl;
R2 is acetoxy or (1-methyl-1H-tetrazol-5-yl)thio, and R3 is carboxy; or R2 is a group of the formula:

Description

This invention relates to a novel cephalosporin compound and processes for preparing the same. More particularly, it relates to a compound of the formula:

4N ~ C--CONH ~ ~ 2 \ O\ N ~ CH -R
CH R
( C~ 2 ) O = C--N
Il wh~rei~ Rl is hydrogen or lower al-~yl;
R2 is acetoxy or (l-methyl-lH-tetrazol-S-yl)thio, and R3 is carboxy; or R is a yroup of the Eormula: ~N~ R ls -COo and Y is hydrogen, hydro.~ymethyl or carbamoyl; and ~ n is an integer o~ 2 or 3, '~

or a pharmaceutically acceptable salt thereof. A pharma-ceutical composi-tion for ~se as an antimicrobial agent which comprises the cephalosporin compound (I) or a salt thereof as an active ingredient together with an inert carrier therefor is also provided in the present invention.
The new cephalosporin compound (I) of the present invention and a pharmaceutical salt thereof show potent antimicrobial activity agains-t a wide variety of microorganisms includiny gram-positive and gram-negatlve bacteria and are useful as anti-bacterial agents, as nutritional supplements in animal food or as chemotherapeutic agents in warm-blooded animals, including man, in the treatment of infectious diseases caused by said gram-positive and gram-negative bacteria.
Among the compounds of the present invention, a preferred subgenus includes those of -the formula (I) in which Rl is hydrogen, lower al'~yl such as me.thyl, ethyl or propyl; R2 iS
acetoxy or (l-methyl-lH-tetrazol-5-yl)thio, and R3 is carboxy;
or R is a group of the formula: - N ~ , R is -COO and Y
2n ls hydrogen, hydroxyme-thyl or carbamoyl; and n is an intec3er of 2 or 3. More 2reEerred subgenus includes those of the Eormula (I) in which R i5 hydroyen or methyl, R2 is (l-methyl-l~l-tetrazol-5~yl)thio, ancl R is carboxy; or R i9 a group of the formula: - N/'~, R is -COo ancl Y is hydrocJen, ~-hyclroxymethyl, 3-hydroxymethyl or 4-carbamoyl; and n is an integer of 2 or 3. E'urther preferred subgenus are those of ~3~

the formula (I) in which R is hydrogen, R is a group of the formula: - N ~ , R is -COo , Y is hydrogen and n is the integer of 2. Most preferred compound is the levorotatory isomer of the compound (I) in which R1 is hydrogen, R2 is a group of the Eormula: - N ~ Y, R3 is -COo , Y is hydrogen and n is the intecJer of 2. In addition, the structural formula (~) shown ahove intends to show that the isomeric configuration of the oxyimino group is Z (i.e., syn)-configuration.
Although the Z (i.e., syn)-isomers of the invention are preferred and show the best biological properties, they may coexist with small amount of E (or anti)-isomer which may exist due to isomerization during the chemical preparation.
According to the present invention, the cephalosporin compound (I) is prepared by the steps of (i) condensing a compound of the formula:

C CO H
O~ (II) C ~1 -tCH2 ) O--c--e~
Rl wherein R is a protecting group ancl R and n are the same as defirlecl abov~, or a r~active derivatLve thereo~ with a compound ~ the ~ormula:

~3~

NH2 ~ S ~ 2 O= - N ~ C~12-R (III) wherein R5 is carboxy or a protected carboxy when R2 is ~cetoxy or (l-methyl-lH-tetrazol-5-yl)thio; or R is -COo when R is a group of -the formula: - N ~ (Y is the same as defined above), or a salt thereof to give a compound of the ~ormula:

R4-NH-~N ~ ICI--CONH~D--~ ~ 2 CH2-R (IV) CH R
(CH2)n O =C - N

wherein Rl, R2, R4, R5 and n are the same as defined above, and (ii) rernovinc3 the protecting group or groups therefrom.
Al~ernatively, the ce2halosporin compound of the formula:

~N --~ C'--CONEI~_-r- ~ ~ C ~ R6 S - ~ CE~ ~ 3 2 (I-a) (CH2 ) O-~C--N
RL

~3~

wherein R is (]-methyl-1H-tetrazol-5-yl)thio or a group of the forrnula: -+ ~ and R , R , Y and n are the same as defined above, is prepared by the steps of (i) reacting a compound of the formula:

~ S~

\S---M O=--N ~ -CH2OCCCH3 (V) CH COOH
(CH2) O=C- N
Rl wherein R is hydrogen or a protecting group and Rl and n are the same as defined above, or a salt thereof with a pyridine compound of the formula: N ~ (Y is the same as defined above),(l-methyl-5-rnercapto)tetrazol or a salt thereof, and (ii) when R is a protecting group, further removing said protecting group therefrom.
In the above-mentioned reacti.ons, a wide variety of protecting cJroups which have been usually employecl to protect amino cJrou2 in the peptide synthesis can be used as the prnte~CtincJ group R or R . Examplcs oE such protectirlcJ
~roups includc lower alkarloyl such as Eo:rmyl, acetyl and p:iv~lloyl; mono-, cli- or trihalogeno-lower alkclnoyl such as chloroacet.yl and trifluoroacetyl; lower alkoxycarbonyl such as methoxycarbonyl, etho.Yycarbonyl and tert.-buto.Yycarbonyl;

3L~3~

substi-tuted or unsubstituted benzyloxycarbonyl such as benzyloxycarbonyl and p-methoxybenzyloxycarbonyl; substituted or unsubstituted phenyl-lower alkyl such as p-methoxybenzyl and 3,~-dimethoxy-benzyl; and di- or triphenyl lower alkyl such as benzhydryl and trityl. On the other hand, when R
in the compound (III) or (IV) is a protec-ted carboxy, the protectiny group on the carboxy group should be -the one which can be easily removed by conventional manners such as hydrolysis, acid treatment or reduction. Examples of such p~otecting yroups include lower alkyl such as methyl, ethyl or tert.-bu-tyli substituted or unsubstituted phenyl-lower alkyl such as benzyl, p-methoxybenzyl and p-ni-trobenzyli benzhydryl; tri-lower alkylsilyl such as trimethylsilyl; and the like. When R is carboxy, i-t is preferred -that -the compound (III) is converted to a salt thereof prior to carryi~ng out the condensation reaction. Suitable examples oE the salt of each one of the compound (III), the compound (V) and (1-methyl-~-mercap-to)tetrazole are inorganic salts such as sodillm, potassium salts or organic salts such as tr;rnethylarnirle, triethylamine salts. Moreover, while the cornpound (I[) can e.Yist in the form of two optical isomers due to the asymmetric car~on atom involved in the ~roup oE
the Eormula:

~ ~ C O ~

~3~

(wherein the asterisk denotes an asymmetric carbon atom ), either an optical isomer of the compound (II) or a racemic modification -thereof may be used for the purpose of the prcsent invention. Throughout the specification and claims, "le~orotatory isomer" of the compound (I), (II), (IV) or (V) :in ~lhlch R is hydrogen and n is an integer of 2 means -that T,he absolute configuration of said compound at said asymmetric carbon atoms is S-configuration and also "dextrorotatory isorner" means that the absolute configura-tion of said compound at said asymmetric carbon atom is R-configuration.
The condensation reaction of the compound (II) or a reactive derivative thereof with the compound (~II) or a salt thereof can be accomplished in conven-tional manners.
For example, the condensation of the compound (II) in its free form with the compound (III) is conducted in the presence of a dehydrating agent in a solvent. Suitable examples of the dehydrating agent include dicyclohexylcarbodiimide, N-cyclohexyl-N'-morpholinocarbodiimide, N-ethyl-N'-(3-dimethyl-aminopropyl)-carbodiimide, phosphorus oxychloride, phosphorus trichloride, thionyl chloride, oxalyl chloride, triphenyl--yhosphine and the like. Vilsmeier reagent prepared from di.m~thylformamide and phosphorus oxychloride, from dimethyl--formamicle and oxalyl chloride,from dimethylformamide and pllosc3ene or from dlmethylformamide and thionyl chloride may al~o be used as said dehydrating agent. It is preferred to carry out the reaction at a temperature of -50 -to 50C, ~ ~3.~

especially -30 to 20C. ~ioxane, tetra-hydro~uran, acetonitrile, chlorofoxm, methylene chloride, dimethylrormamide, N,N-dimethyl-acetamide, ethyl acetate, pyridine, acetone and water are suitahle as the solvent.
On the other hand, the condensation reaction of the reactive derivative o~ the compound (II) with the compound (II:L) or a salt thereof can be conducted either in the presence or absence of an acid acceptor in a solvent.
Suitable examples of the reactive derivative of the compound ~0 (II) include the corresponding acid halides (e.g., chloride, bromide), mixed anhydrides (e.g., a mixed anhydride of the compound (II) with alkyl carbonate~, active esters (e.g., p-nitrop'nenyl ester, 2,~-dinitrophenyl ester, succinimide ester, phthalimide ester, benzotriazole ester, 2-pyrrolidon-1-yl ester), acid azide and acid amides (e.g., imidazole amide, 4-substitu-ted-imidazole amide, triazole amide). Dioxane, tetrahydrofuran, acetonitrile, cnloroform, methylene chloride, dimethyl~ormamide, N,N-dime-thylacetamide, ethyl acetate, pyridine, acetone and water are suitable as the solvent.
Moreover, suitable examples of the acid acceptor include alkali metcll hydroxides (e.g., potassium hydroxide, soclium hydroxide), alkaLi metal carbonates or bicarbonate~ (e.g., sodillrn carbona~e, sodium bicarbonate, potassium carhonate, potassium bicarbonate), trialk.yl amines (e.c3., trilnethylamine~, kril-~thylamine), N,N-dial~ylanilines (e.g., N,N-dimethylanilinc~, N,N-diethylaniline), pyridine and N-alkyl-morphorines (e.g., ;~3~

N-methylrnorpholine). It is preferred to carry out the reaction at a temperature of -50 to 50C, especially at -30 to 20C.
The removal of the protecting group or groups from the compound (IV) thus obtained can be conducted by conventional manners such as, for example, hydrolysis, solvolysis, acid treclttnent or reduction. For example, when the protecting group R is formyl, acetyl, tert.-butoxycarbonyl, benzhydryl or trityl and the protecting group on the carboxy group is tert.-butyl or benzhydryl, said group or groups may be removed by treating the compound ~IV) with an acid. Suitable examples of such acid include, for example, formic acid, trifluoroacetic acid, benzenesulfonic acid, p-toluene-sulfonic acid, hydrochloric acid or hydrogen bromide, especially trifluoroacetic acid. This reaction may be conduc-ted with or without a solvent. Examples of the solvent àre water, methanol, ethanol, acetic acid or dioxane. It is preferred to carry out the reaction at a temperature of -30 to 70C, especially 0 to 30C. Moreover, when the trifluoroacetic acid is used as the acid, it is preferred to carry it out in the presence of anisole. When the pro~ectinc3 group R is henzyloxycarborly], p-met'noxybenzyloxycarbonyl, benzyl, p-metho:~y-benz~l or 3,4-d:imethoxyherlzyl and the protecting group on the carboxy group is benzyl, p-methoxybenzyl or p-ni.trobenzyl, the removal of said protectinc3 group or grvupC; may be conclucted by catalytic h~,~drogerlation of the compound (IV) in hydrogen gas in the presence of a catalyst.
This catalytic hydrogen-ation is preferably carried out at a temperature of 0 to 100C, especially 10 to 40Cr under atmospheric or increased pressure. Preferred examples of the catalyst include palladium BaCO3, palladiurn-charcoal and palladium-black. Methanol, ethanol, tetrahyclroEuran and w~ter are su:itable as the reaction solvent. Further, when khe protecting group R is trifluoroacetyl, pivaloyl, methoxy-carbonyl or ethoxy-carbonyl and the protecting group on the carbo~y group is methyl or ethyl, said group or groups may be removecl by hydrolysis of the compound (IV). The hydrolysis of the compound (IV) can be carried out in conventional mahners, for e~ample, by treating it with an alkali agent such as sodiurn hydroxide or potassiurn hydroxide, or an acid such as hydrochloric acid or hydrobromic acid. It is preferred to carry out said hydrolysis at a tesnperature of 0 to 70C, especially 10 to 30C. When the protecting group R4 is chloroacetyl, said group may be removed by treating the compound (IV) with thiourea in a solvent. ~ethanol, ethanol and water are suitable as the solven-t. It is preferred to carry it out at a ternperature of 20 to 80C, esp~cial]y 40 to 80C.
On the otsler hancl, the reactlon of the compourld (V) or a scllt khereoE with the pyridine compound N/'~) or (l-rnethyl-5-mercapto)-telra~ole or a salt thereof can be reaclily conduct:~d in a solvent. Water, dimethylforlllarnide an --ll--N,M-dimethylacetamide are suitable as the solvent. It is preferred to carry out the reaction a-t a temperature of 0 to 100C, especially 20 to 80C. It is also preferred to carry it out in -the presence of sodium iodide, potassium iodide, sodium bicarbonate or phospha-te buffer solutiGn.
RernovaL oE the protecting group R of the product thus obtalned can be conducted in the same manner as in removing the protecting group or groups from the compound (IV).
Concomitantly, the starting compound (II) of the present L0 invention is prepared, for example, by reacting a compound oE the formula:

1 r S - N (VI) - OH
wherein R is the same as defined abo~e, with a compound oE the formula: X-CH 2 n N-R ~wherein X is halo~en and R and n are-the same as defined above) in the presence oE an alkali agent (e.g., potassium carbonate) in a solvent (e.g., dimethylsulfoxide) at a temperature of 10 to 50C to give a compound of the forrnula:

-~3~

4 ~ N ~ C- C2C2H

S N\ o (vII) CH
(CH2) O =C - N
Rl hereln R , R ancl n are -the same as defined above, and then hydrolyziny the compound (VII). Alternatively, the starting cornpound (II) may be prepared by hydrolyzing the compound (VI) -to give a compound of the formula:

~ //N ~-- C-- C02H
R -NE-~
\ S - ~OH (VIII) wherein R is the same as defined above, and then reacting the c~npound (VIII) with a compound of the formula: X-C~ ~ 2 n N-R (wherein R , n and X are the same as defined above) in the presence of an acid acceptor (e.y., sodium hydride) at a temperature of 10 to 40C in a solvent (e.y., dimethylsulfoxide). Moreover, as mentioned hereinbefore, the compound (II) involves two optical isomers due to the asylnmetrLc carhon involved in the group of the Eorlnula: -C*EI 2) ~ N-R (wherein the asterisk denotes ~ CO~
an asymmetric carbon atom). If required, however, such optical isomers may be separated into eaeh optical isomers by optical resolution thereo~. For example, the eompound (II) in which R is hydrogen, n is an integer o~ 2 and R4 is tr;.tyl can be readily separated into eaeh opti.eal isomers by reactiny the racemie modiLieation o~ the eompound ~II) with L~ or ~-phenylalanine methyl ester in a solvent (e.g., a mi~ture of methanol and dioxane) to form the diastereoisomerie salts thereo~, and separating said diastereoisomers into each eornponents thereof by selective reerystallization. By said selective recrystallization, the least soluble diastereo-isomer is reeovered as erystals from the reaetion mi.Yture and the more soluble diastereoisomer remains soluble therein.
It is preferred to carry out said seleetive crystallization at a tempera-ture of 10 to 40C.
The eephalosporin eompound (I) of the present irlvention and a pharmaeeutically aeeeptable salt thereof show potent antimicrobial aetivity against.a wide variety of mieroorganisms including those belonging to th~ genera Streptocoecus (e.g., St. faeealis, St. pneumoniae); Staphyloeoeeus (e.g., S.
aure-ls, S. epiclermidis) and Pseudomonas (e.g., Ps. aeruc3inosa, Ps. putida, Ps. .stut-zeri), and are particularly charaeterizecl by their poten-t antimicrobial ac-tivity against both ~3ram-positive and qrclm-negative bact~ria. For example, 7B-~(Z)-2-(2-arni.no-thi.azol-4-yl) 2-((2--pyrrolidon-3-yl)o~yirnino)clcetamido3-3-(l-pyrldilliomethyl)-3-cephell~ -carbo.~ylate (~,-isorner) and 7B-~(Z)-2-(2-aminothiazol-4-yl)-2-~(1-methyl-2-pyrrolidon-3-yl)-3~

o~yimino acetamldo}-3-l(l-methyl-lH-tetrazol-5-yl)thiomethyl~-3-cephem-4-carboxylic acid exhibit minimum inhibitory concent-rat;on (.~.I.C.) (Agar dilution method, cultured for 20 hours at 37C) of 12.5 and 25/~g/ml against Streptococcus faecalis CN ~7B, while the M.I.C. of Cefmenoxime ~Chemical name.

7~ )-2- (2-aminothia~ol-~-yl ) -2- (methoxyimino)acetamiclo) -3-~(L-methyl-lH-tetra~ol-5-yl)thiomethyl~-3-cephem-~-carboxylic acid)arld CeEta~iditne (Cllelnical name: 7B- ~(Z)-2-(2-aminothiazol-~-yl)-2-(2-carboY.yprop-2-yloxyimino)acetamido)-3-(1-pyridinio-rneth~l)-3-cephem-4-carboxylate~ against said microorganism are more than 100~/lg/ml. The antimicrobial activl-ty of 7B-{(Z)-2-(2-atninothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oxyimino~-acetamido}-3-~ methyl-lH-tetrazol-5-yl)thio-methyl~-3-cephem-4-carboxylic acicd against Staphylococcus aureus 252R is also more than 16 times stronger than those of Cefmenoxime and Cefta~idime. Moreover, the compound (I) and a salt thereof show potent antimicrobial activity against bacteria belonging to the genera Bacillus (e.g., B. subtilis), Escherichia (e.g., E. coli), Klebsiella (e.g., K. pneumoniae), Enterobacter (e.g., E. aerogenes, E:; cloacae) and Serratia (e.g., S.
marcescerls ) .
rL'hc-~ ce~halosporin compouncl (I) may furthcr e~hiblt pol:ellt ant:imicrobial activity against ot~er b.lcteria beloncJillc3 to the genera Citrobclcter, Protells, Shigella, l~etllopllilus and Salmc)nella. Further, the cephalosporin compound (I) ancl a salt thercof are characteri~ed in that they show potent :~3~

pro-tective effects against microbial infections of various bacteria including both Staphylococcus aureus and Pseudomonas aeruginosa, because of its high absorbability or long-lasting therapeutic effects in living tissues; that they have a high stabllity against a varity of B-lactamase-producing micro-oxganisrns, especially against 3~1actamases produced by Prokeus vulyaris; and also that they are low in toxiclty.
For example, no rats die even after 7B-l(Z)-2-(2-aminothiazol-~-yl)-2-~(2-pyrrolidon-3-yl)o~yimino3acetamido}-3-~1-pyridinio-methyl)-3-cephem-4 carboxylate (~-isomer) is administered subcutaneously to SD-male rats at a dose of 1000 mg for 14 consecutive days.
The cephalosporin compound (I) of the present invention can bè used for pharmaceutical use either in the free form or in the form of a salt thereof. Pharmaceutically acceptable salts of the compound (I) inclu~e, for exrample, non-toxic me-talic salts such as sodium, potassium, calcium or aluminum salts; ammonium salt; salts thereof with non-to~ic amines such as -trialkylamines (e.g., triethylamine and procaine);
salts thereof with inorganic acids such as hydrochlori.c acid or hydrobromlc acid; salts thereof with organic acids such as ox.ll;c aci.d or tartaric acid; and so forth. These salts r-lr(.~ f.`a~ ly obtalne~d by treatinc~ the compourld (I) with a stoi~:h:iolnetr:ical]y equi-molar amount of the corresponding alkali ac~erlt or acid at around room temperature in an aqueous ~qolv~nt. Th~ cephalosporin compound (I) and a salt thereof can be administered either orally or parenterally (e.g., intravenously, intramuscularly, subcutaneously). The daily dose of the compound (I) or a salt thereof may vary over a wide range depending on -the age, weight or conditions of pat;ents, and the severity of diseases to be treated. In genercll, ho~ever, a preferred daily dose of said compound ~I) or a salt thereof may be a'~out 0.002 to about 0.2 g, especially 0.01 to 0.04 g, per kg oE body weight per day.
Further, the compound (I) and a salt thereof may be used in the form o~ a pharmaceutical preparation containing the same compound in conjunction or admixture with pharmaceutical exclpients suitable for oral or parenteral administration.
Suitable excipients include, for example, gelatin, lactose, glucose, sodium chloride, starch, magnesium stearate, talcum, vegetable oil and other kno~n medicinal excipients. The pharmaceutical preparations may be in solld form such as tahlets, coated tablets, pills or capsules; or in liquid form such as solutions, suspensions or emulsions. They may b~ sterillized and/or may furt}-er contain auxiliaries such as stabilizing, wetting or emulsifying agent.
Pract:ical and presently-preferr2d emhocliments o~ the prese~llt inv~ntion are illustratively sho~n in the followincJ
E'xamples. Throughout the specification and claims, the t~rm "lower alkyl" should be interpretecl as referrinc] to al~;yl havlng one to four carbon atoms.

~3~

Experiment I
(Antimicrobial activity in vitro) ~ rhe minimurn inhibitory concentration (MIC,~ug/ml) of a test cornpouncl was determined by means of standard agar plate dllution methocl (based on the standard method of Japarl Society of Chemotherapy). Media used in this e.Yperiments w~re Mueller~~linton agar (MHA; Niscui).
The results are shown in the following Table 1.

~23~

Table 1 M. I. C. ( q/ml) MicroorganismsThe compound of the Cefmenoxme ~ -tested (Note- 1) (Note 2) Ce tazld me Staphylococcus aureus 0.781.56 l2.5 Tera~ima _ . . _ _ . . _ _ _ _ . _ . . _ _ _ _ staphylococcus aureus 25 ~ 100 > 100 streptococcus faecalis 12.5 > 100 > 100 -Bacillus subtilis 0.2 1.56 3.13 Klebsiella pneumoniae 0,05 0.1 0.1 _ Enterobacter cloacae ~ 0.05 , 0.1 0.2 Serratia ma6rcescens < 0 05 0.2 ~ 0.2 _ _ .
pseudomonas aeruginosa 0.39 6.25 0.78 _ _ _. . . _ . . _ . . . _ . . _ _ ATCC 12633 1.56 100 12.5 . _ . . . . . . _ _ . _ .
Note~ 7r~- {(Z)-2-(2-aminotniazol-4-yl)-2-((2-pyrrolidon-3-yl)-oxyimino)acetamido}-3-(1-pyridiniomethyl)-3-cephem-~-carboxyklte~ isomer)

2): Chemical name = 7B-~(Z)-2-(2-aminothiazol-~l-yl)-2--metho.~yLmino)clcetalllido)-3-((l-methyl-l~I-tetrazol-5-yl)-thiomethyl)-3-cephem-~-carbo~ylic acid

3): Chenlical name - 7B-~(Z)-2-(2-aminothiazol-~1-yl)-2-(2-carbo~y-prop-2-ylocyimino)acetamido)-3-(1-pyridinionlethyl)-3-cephem-4-carbo.Yylatc 3~

Experiment I
(Protective effects on bacterial infections in mice) Ten male mice weighing 20 + 1 g were used for each dose level. Mice were challenged via the in-traperitoneal route Witil su~ficlent ~acteria to kill all non-treated mice within 2~ hours. All bacteria were suspended in 6 ~ mucin.
test compound were administered intramuscularly one hour aEter the infection. Survival ratios were determined 7 days aEter the infection. ~he median effective doses (ED50, mg/kg) of the test compo-und was estimated by the probito analysis.
The results are shown in the following Table 2 together with the M. I. C. (~g/ml) of the test compound which was estimated in the same manner as described in Experiment I.

3~3~

Table 2*

.
ED50 (mg/kg ) Mic~oorc3~nismsThe compound of the tested present invention Cefmenoxlmf~ Ceftazldlme Stclph~lococcus aureus 1.71 5.32 7.85 S~ni~h (1.56) (1.56) (12.5) ~schFrLchia coli 0.05 0.16 0.08 KC-14 (0.05) (0.025) (0.05) ., _ .
Serratia marcescens 0.14 0.88 0.5 7006 (0.05) (0.2) (0.2) citrobacter freundii 0.06 0.19 0.15 916 (0.1) (0.05) (0-39) .
Enterobacter aerogenes l.. 37 23.85 26.38 816 (0-39) (1.56) (6.25) -ote: * : the numerical values in parenthesises show the M. I. C.
(minimum inhibitory concentration, ,ug/ml) of each test compounds.
1) - 3~ : same as shown in the footnote of Table 1.
e _ (1) 3.2 cJ oE (Z)-2-(2-tritylaminothiazol-4-yl)-2-~ (2-pyrrolidon-3-yl)oxyimino~acetic acid are suspended in 60 ml of tetrahydrofuran, anci 2.05 g of tert.-butyl 7-aminocephalo-sporcln.lte, 1.27 g of l-hydroxybenzotriazole and 1.~3 g oE

clicyclohexylcarbodiimidf are added thereto. The mixture is st-irrecl at room tfmpfrature for 3 hours. Insoluble materials ar.f~ Eiltered ofE, and the Eiltrate is concentrated to dryness ~Itld(~r rc~duced pressure. The residue is dissolved in ethyl aeetate, and the solution is washed with 1 ~ hydroehlorie aeid, 5 % aqueous sodium ~iearbonate solution and water, suceessively. The e-thyl aeetate solution is dried and coneentrated to dryness under redueed pressure. Then, the ~es:lclue thus obtained is puriEied by siliea gel ehromatography (solvent, ehloroforrn: methanol = 98.5: 1.5). 4.3 y of tert.-butyl 7~-~(Z)-2-(2-tritylaminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oxyirnino)aeetamido}eephalosporanate are obtained as pale yello~,l powder.
M.p. 135 - 145~C (deeomp.) NMR (CDC13)S: 1.52 (9H, s), 2 02 (3H, s), 2.2 -2.7 (2H, m), 3.0 - 3.5 (4H, m)

4.5 - 5.3 (~H, m), 5.6 - 6.0 (lH, m) 6.70 (lH, s), 6.9 - 7.5 (17ri, m) 8.4 - 8.7 (lH, broad) (2) 1.0 g of tert.-butyl 7B- ¦ ( Z ) -2-(2-tritylaminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oxyimino)aeetamido3eephalosporanate is acidecl to a mixture of 20 ml of trifluoroaeetie aeid and one ml oE anisole, and the mixture is stirred at room tempe~r-ature for 20 mirlutes. The mixture i5 eoneentrated under dlle~?d prc?ssurc? to r~?movc? trifllloroaeetie ~leid. Ether is adclecl to thc! residue, and the resultinc3 powder is collected l)y Liltratiorl. The powder is suspended ln 10 ml of water arlcl soclium biea~bonate is added thereto to dlssolve said L)owder there~in rrhe solution is ~ashed with ethyl aeetate and ehrc)lTIatoc~raplled on a eolumn of non-ionie polymer res;rl -~3~9~

Amberlite XAD-2 (resistered trade mark, manufactured by Rohm & Haas Co., ~.S.A.), using water as an eluent. The frac-tions containing the cephalosporin compound are collected and concentrated under reduced pressure to remove solvent.
Then, acetone i.s added to the residue obtained, and the resulti.ny powder is collected by filtration. 320 mg of sodi.um 7B~ 2-(2-aminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oxyi.mino~acetamido}cephalosporanate are obtained as colorless powder.
NMR (~2)~ 2.10 (3H, s), 2.2 - 2.7 (2H, m) 3.15 - 3.85 (4H, m), 4.6 - 5.00 (2H, m)

5.01 (lH, t, J= 7Hz), 5.16 (lH, d, J= 5E~z) 5.77 (lH, d, J= 5Hz), 6.98 (lH, s) Example 2 (1) 4.0 g of (Z)-2-(2-tritylaminothiazol-4-yl)w2-~(2-pyrrolidon-3-yl)oxyimino~acetic acid are dissolvèd in a mixture of 30 -nl of tetrahydrofuran and 10 ml of N,N-dimethyl-acetamidc, and 1.27 g of l-hydroxybenzotriazole and 1.93 g of dlcyclohexylcarbodiimide are added thereto. After. the rnixture is stirred at 0 to 5C for 2 hours, said rnixture is addecl to 30 rnl of N,N-dimethylacetamide-water (water content:
lS 'k) corltai.nirlcJ 2.12 r~ of 7-arni.nocephalosporanic acid and 4 rJ of triethylamille. Said acldition i.5 carri.ed out under i.cc-cool:in~J. Then, the m:ixture is stirxed at the same tenlper,ltUre for 1.5 hours. Insoluble materi,-lls are filtered off, and thc filtratc is concentrated under recluced pressure to remove solvent. The residue is poured into 300 ml o~
water. Said mixture is adjusted to pH 8 with sodium bicarbonate, washed with ethyl acetate, adjusted to pH 3 with 2N hydrochloric acid, and then extracted with ethyl acetate. The extract ;.s dri.ed and concentrated to dryness under reduced pressure.
Ether is added to the residue thus obtained, and the resul-ting pow(ler is collected by flltration. 3.1 g of 7~-~(Z) 2-(2-tritylarninothiazol-4-yl)-2-((2-pyrrolidon-3-yl)oxylmino~-acetamido3cephalosporanic acid are obtained.
~R (DMSO-d6)~: 2.03 (3H, s), 2.1 - 2.5 (2H, m) 3 0 - 3.7 (dH, m), 4.4 - 5.2 (4H, m) 5.5 - 5.9 (lH, m), 6.71 (1~, s) 7.0 - 7.6 (lSH, m), 7 84 (lH, s) 8.80 (lH, broad s), 9.50 (lH, broad d) (2) 40 ml oE 80 % aqueous formic acid are added to 3.0 g of 7B-~(Z3-2-(2-tritylaminothiazol-4-yl)-2- ~(2-pyrrolidon-3-yl)oxyimino)acetamido¦-cephaIosporanic acid, and the mixture is stirred at room temperature Eor 2 hours. Insoluble mate~rials are Eilte.red off, and the filtrate is concentrated to dryness under reduced pressure. Water is aclded to the residue, arld the aqueous mi:cture is rleutrali~ed with sodium bicarbon.lte ancl washed with ether. Then, said aqueous m~.ture is chromatographed on a column oE norl--ionic polymer r.esin D1.aiorl ~IP-20 (rcsistered trade mark, marlufactured by ~itsubisili Chemical Industries Ltd., Japan), using water as an eluerlt. The fractions containirlg the ce~phalosporin compound 3~

are collected and concentrated to dryness under reduced pressure. 1.5 g of sodium 7~-¦(Z)-2-(2-aminothiazol-4-yl)-2 ~(2-pyrrolidon-3-yl)oxyimino)acetamido 3 -cephalosporanate are obtained.
The physico-chemical properties of this product are ntical ~/ith those of the sample obtained in Example (2).
~X1nl~
(1) 3.25 g of (Z)-2--(2-tritylaminothiazol-~-yl)-2- ~(2-pyrrolidon-3-yl)oxyimino)acetic acid are dissolved in 200 ml of tetrahydrofuran, and 3.14 g of benzhydryl 7B-amino-3- ((1-methyl-lH-tetrazol-5-yl)thiomethyl~-3-cephem-4-carboxylate, 1.03 g of 1-hydroxybenzotriazole and 1.57 g of dicyclohexyl-carbodiimide are added thereto. The mixture is s~irred at room temperature for 2 hours. Insoluble materials are filtered off, and the filtrate is concentrated to dryness under reduced pressure. The residue is dissolved in ethyl acetate, and washed with 1 ~ hydrochloric acid, 5 ~ aqueous sodium bicarbonate solution and an ayueous saturated sodium chloride solution, successively. The ethyl acetate solution is th~.?n dried and concentrated to dryness under reduced pr~?ssur~?. T~i~? r~?sidu~ t~lus obtain~?d is purifiecl by silica ~el chrolnatography (solvent, ethyl acetate: benzene =
S: 2). 3.7 ~ o~ benzhydryl 7~-~(Z)-2-(2-trit:ylalnirlothiazol-~-yl)-2-~(2-pyrrolidon-3-yl)o~yimillo)aci?tarn;doJ-3~((l-methyl-~ tetrl-z.ol-S-yl)thiorrl~-?t:hyl)-3-cephern-~-carbo.~ylate are obtained.

3~

M.p. 122 - 126C (decomp.~
NMR (CDCl3)(: 2.2 - 2.7 (2H, m), 3.0 - 3.5 (2H, m) 3.6 - 3.75 (2H, m), 3.78 (3H, s~
4.2 - 4.4 (2H, m), 5.03 (2H, m) 5.7 - 6.05 (lH, m), 6.75 (lH, s)

6.88 (lH, s), 7.1 - 7.5 (27H, m) 8.80 (lH, broad) (Z) 360 mg of benzhydryl 7B-~(Z)-2-(2-tritylaminothlazol-9--yl)-2-~(2-pyrrolidon-3-yl)oxyirnino~acetamido~-3-~(1-methyl-1~ lrI-tetrazol-5-yl)thiomethyl~-3-cephem-4-carboxylate are added to a mixture of 10 ml of trifluoroacetic acid and 0.5 m] of anisole, and the mixture is stirred at room temperature for 20 minutes. The mixture is concentrated under reduced pressure to remove trifluoroacetic acid. Ether is added to the residue, and the resulting powder is collected by filtration.
The powder is suspended in water, and sodiurn blcarbonate is added thereto to dissolve said'powder therein. The solution is washed with ethyl acetate and chromatographed on a column of noI-l-ionic polymer resin Amberlite XAD-2 (resistered trade mark, marlufactured by Rohm & EIaas Co., U.S.A.), using water as an eluent. l'he fractions containing the cephalosporin compound are collected and concentrated to dryness under redllced prc~ssure. 0.3 g o~ sodium 7B-~(~)-2-(2-aminothiazol-~I-yl)-2-((2-pyrrolidon-3-yl)oxyimino)acetamido}-3-~(1-rnethyl-lII-tetrazol-S-yl)thiomethyl~-3-cephem--1-carboxylate is obtainecI.

(DMSO-d63~ 2.1 - 2.5 (2H, m), 3.1 - 3.5 (2H, m) 3.94 (3H, s~, 4.2 - 4.5 (2H, m) 4.6 - 4.8 (lH, m), 5.03 (lH, d, J= 5Hz) 5.5 - 5.8 (lH, m), 6.76 (lH, s)

7.3 (2H, broad s), 8.00 (lEI, s) 9.SS (lH, broad) ple ~
0.57 y oE oxalyl chloride is added at -5 to 0C to 15 ml of chloroform containing 0.35 g of dimethylformamide, and the mixture is stirred at the same temperature for 15 minutes.
A mixture of 1.54 g of (Z)-2-(2-tritylaminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oxyimino3acetic acid, 0.3 g of -triethylamine and 15 ml of chloroform is added to said mixture at ~ 30 to - 35C, and the mi~ture is further s-tirred at the same temperature for 5 rninutes. Then, a solution of 73-amino-3-(1-pyridinio-methyl)-3-cephem-4-carboxylat2 in chloroform (said solution is prepared by suspending 1.82 g of the dihydrochloride of said cephem compound in 10 ml of chloroform and ad~ing 4 ml of N,O-bis(trimethylsilyl)acetamide thereto to dissolve said salt therein) is added to said mi~ture at -35~ to - 30C, and the mixture is stirred at the same temperature ~or 10 minutes and at ~ 30 to - 10C for one hour. Thr mi.cture is concentrated to dryness under reduced pressure.
~0 rlll oE 80 ~ aq~leo-lC; Eormic acid are added to the residue, ancl the aqutous micture is stirred at room telnperature for one hour. 50 rnl of water are added to said ~Iqueolls mixture.

Then, insoluble materials are filtered off, and the filtrate is washed with ethyl acetate and concentrated to dryness under reduced pressure. The residue thus obtained is dissolved in wa-ter and chromatographed on a column of non-ionic polyrner resin Diaion HP-20 (resistered -trade mark, manufactured by Mitsuhishi Chemical Industries Ltd., Japan). The column is ashed with w~ter, followed by elution with 20 ~ methanol.
~rhe fractions containing the cep'nalosporin compound are collected and concentrated to dryness under reduced pressure.
:L0 Acetone is added to the residue thus obtained, and the resulting powder is collected by filtration. 0.84 g of 7B-~(Z)-2-(2-aminothiazol-4-yl)-2- ~(2-pyrrolidon-3-yl)oxyimino)-acetamido3-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate is obtained.
M.p. > 250C
NMR (D2O)~: 2.2 - 2.7 (2H, m), 3.1 - 3.7 (4~, m) 4.9 - S.S (4H, m), 5.80 (lH, d, J= SHz) 6.92 (1~, s), 7.8 - 9.1 (5rl~ m) ~20 ~ 13 4(C= 1.0, ~l2O) Exln~le 5 1.56 cJ of oxal~l chloride are added at - 5 to 0C to 39 ml of chloroform contai.nincJ 0.99 ml of dimethylformamide, and the m:ixt~lre is stirred at the same temperature for lS
minutes. ~ solutiotl of 4.23 g of (Z)-2-(2-tritylaminothiazol-~-yl)--2-((2-pyrrol;don-3-yl)oxyirnino)acetic acid (d-isomer) an(l 0.8-~ ~ oE triethylamine in 39 ml of chloroEorrn is addecl to said mixture at - 30C. The mi~ture is stirred at the same temperature for 5 minutes. Then, a solution of 7B~amino-3-(1-pyridiniomethyl)-3-cephem-4-carboxyla-te in chloroform ~said solution is prepared by suspending 5.0 g of the dihydro-chloride of said cephern compound in 39 ml of chloroEorm and addincl 11 ml of N,O-b:is(trime-thylsilyl)acetamide thereto to dissol~ said salt therein) is added to the said mi~ture a~ - 30 -to - 10C. After the mixture is stirred at the same temperature for 30 minutes, said mi,c-ture is concentrated to dryness under reduced pressure. 100 ml of 80 ~ aqueous formic acid are added to the residue, and said aqueous mixture is stirred at room -temperature for one hour. 100 ml of water are added to the mixture, and insoluble rnaterials are filtered off. The filtrate is washed with ethyl acetate and is concentrated to dryness under reduced pressure. The resLdue thus obtained is dissolved in water and chromato-graphed on a column of non-ionic polymer resin Diaion HP-20 (resistered trade mark, manu~actured by ~itsubishi Chemical Industries Ltd., Japan). The column is washed with water, followe-1 by elution wi~h 20 ~ aqueous methanol. The fractions containi.ng the cephalosporin compound are collected and concentrated to dryness under reduced pressure. ~cetone is a(lclc(l to the residue, the resultin~ powder is collected by filtrat:ion. 2.14 g of 713-{(Z)-2-(2-aminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oY~yimino~acetamido¦-3-(1-pyridiniorllethyl)-3-cephem-4-carboxylate (d-isorner) are obtained. ~nother designation oE this deY~trorotatory isomer is shown as 7~-¦(r~)-2-(2-aminothia~ol-4-yl)-2-~((3R)-2-pyrrolidon-3-yl)-oxyimino~ acetamido}-3-(1-pyridiniomethyl)-3-cephem-4-carbox~late.

~3~

.~9 NMR (D2O)~: 2.1 - 2.7 (2H, m), 3.1 - 3.7 (4H, m) 4.9 - 5.5 (4H, m), 5.79 (lH, d, J= 5Hz) 6.92 (1~, s), 7.8 ~ 9.1 (5H, m) ~D ~ 45-7 (C= 1, H o) E~6 1.81 g of oxalyl ehloride are added at - 5 to 0C to ~Sm1 of chloroform containing 1.15 ml of dimethylformamide, and the tnixture is stirred at the same temperature for 15 rninutes. A solution of 4.90 g of (Z)-2-(2-tritylaminothiazol-1~ 4-yl)-2- ((2-pyrrolidon-3-yl)oxyimino~aeetie aeid (~-isomer) and 0.97 ~ of triethylamine in 45 ml of ehloroform is added to said micture at - 30C. The mixture is stirred at -the scime temperature for 5 minutes. Then, a solution of 7~-amino-3-(1-pyridiniomethyl)-3-eephem-4-earboxylate in ehloroform (said solution is prepared by suspending 5.8 g of the dihydro-ehloride of sald eephem eompound in 45ml of ehloroform and adding 12.7 ml of N,O-bis(trimethylsilyl)aeetamide thereto to dissolve said salt therein) is added to the said mi.Yture at ~ 30 to - 10C. After the mi~ture is stirrecl at the ;ame tc~nnperature for 30 minutes, said micture is eoneentrated to dryness under rcducecl pressure. 100 ml of 80 ~ aqueous ~orrnic acicl ar~ adcled to thc residue, and said aqueous IlI:i.'.~tllr-' i5 stirrcd at room temperature for one hour. ].10 ml o~ water are adclccl to thc rni~ture, ancl insoluble rnaterials arc ~i.ltcrcd o~f. The Eiltrate is washed with ethyl aeetate arl(l -is eoncelltrclted to dryness under recluced pressure. The residue thus obtained is dissolved in water and chromato-yraphed on a column of non-ionic polymer resin Diaion ~P-20 (resistered trade mark, manufactured by Mitsubishi Chemical Industries Ltd., Japan). The column is washed with water, Eollowed by elution with 20 '~ ayueolls methanol. The fractions contc~linincJ the cephalosporin compound are collected and concentratecl to dryness under reduced pressure. Acetone is adcled to the residue, and the resulting powder is collected by fi]tration. 2.22 g of 7B-~(Z)-2-(2-aminothiazol~4-yl)-2-~(2-]o pyrrolidon-3-yl)oxyimino)acetamido~--3-(1-pyridiniomethyl)-3-cephem-4-carboxylate (~-isomer) are obtained. Another desiynation of this levorotatOry isomer is shown as 7B-¦(Z)-2-(2-aminothiazol-4-yl)-2-(((3S)-2-pyrrolidon-3-yl)oxyimino)-acetamido}-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate.
MMR (D2O)~: 2.2 - 2.7 (2H, m), 3.1 - 3.8 (4H, m) 5.05 (lH, t, J= 7Hz), 5.28 (lH, d, J= 5Hz), 5.36 (lH, d, J= 14'-lz), 5.63 (lH, d, J=14Hz), 5.87 (lH, d, J= 5Hz), 6,98 (lH,s),

8.10 (2H, t, J= 7.5Hz), 8.S7 (lH, t, J=7.5Hz) 8.98 ~]~-l, d, J= 7.5~1z) ~2 _ 38.0 (C= 1, H2O) ~Y-lmp]~? 7 ~ mi:cturc oE 13 g of sodium iodkl~ and ~I ml o~ water is stirrc-!cl at 30C. 3~6 cJ o~ pyridirle arld 3.2 cJ oE 7B-{(Z)-2-(2-aminothiazol-~1-yl)-2-~(2-pyrroliclon-3-yl)o:;yirnillo~acetamido,}-cephLllosporallic acid are added to said mi.cture, and the ~LZ3~

mixture is stirred at 75 to 80C for one hour. After cooling, the reaction mixture is poured into 150 ml of water and concentrated to dryness under reduced pressure. The residue is dissolved in 150 ml of water, and the solution ls adjusted to pH 1 with 2N hydrochloric acid. Insoluble materials are filtered ofE, and the filtrate is washed with ethyl acetate, adjusted to pH 6 with 2N sodium hydroxide solution and concentrated to a total volumn of 30 ml under reduced pressure. The solution thus obtained is chromato-~raphed on a column of non-ionic polymer resin Diaion ~IP-20 (resistered trade mark, manufactured by Mitsubishi Chemical Industries Ltd., Japan). The column is washed with water, followed by elution with 20 % aqueous methanol. The fractions containing the cephalosporin compound are collected and concentrated to dryness under reduced pressure. Acetone is added to the residue thus obtained, and the resu~ting powder is collected by filtration. 0.67 g of 7~-{(Z)-2-(2-aminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oxyimino~acetamido~-3-(1-pyridinio-methyl)-3~cephem-4-carboxylate is obtained.
The physico-chemical properties of the product are identical with those of the sample obtained in Example 4.
Example 8 (1) 1.5 g of (Z)-2-(2-tritylaminothiazol-4-yl)-2~
methyl-2-pyrrolidon-3-yl)oxyimino)acetic acid are dissolved in 50 ml oE tetrahydroEuran, and 1.4 g of benzhydryl 7~-a[nino-3-~(1-rn~thyl-1~l-tetrazol-5-ylJthiomethyl)-3-cephem-4-carboxylate, 0.6 g of l-hydroxybenzotriazole and 0.92 g of dicyclohexyl-carbodiimide are added thereto. The mixture is stirred at room temperature for 2 hours. Then, the reaction mixture is -treated in the same manner as described in Example 3-(1).
1.5~ g of ben~hydryl 7~-{(Z)-2-(2-tritylaminothiazol-4-yl)-2-~(1-rnethyl-2-pyrrolidoIl-3-yl)oxyimino~acetamido3-3-((l-me-thyl-lfI-tetrazol-5-yl)thiomethyl)-3-cephern-4-carboxylate are obtained.
NMR (CDCl3)~: 2.2 - 2.6 (2H, m), 2.75 (3H, s) :L0 3.1 - 3.5 (2H, m), 3.5 - 3.8 (2H, m) 3.84 (3H, s), 4.2 - 4.4 (2H, m) 4.8 - 5.2 (2'I, M), 5.8 ~ 6.1 (lH, m) 6.72 (lH, s), 6.90 (lH, s) 7.0 - 7.5 (25H, m), 8.5 - 8.8 (2H, broad) (2) 1.5 g oE benzhydryl 73-{(Z)-2-(2-tritylaminothiazol-4-yl)-2- ~(1-methyl-2-pyrrolidon-3-yl)oxyimino~acetamido}-3-~(1-methyl-lH-tetrazol-5-yl)thiomethyl)-3-cephem-4-carboxylate are added to a mixture of 1.5 ml of anisole and 10 ml of trlfluoroacetic acid, and the mixture is stirred at room t~mperature for 30 minutes. The rnixture is concentrated under reduced pressure to remove trifluoroclcetic clcid.
Etilcr i5 added to the residue, and the resulting powder .i5 col]ected by filtr.ltion. The powder is sus;~ended in 15 ml o~ water, and sofliulll bicarhonaté is aclded thereto to dissolve sa;d pow-lcr therein. The so:Lution is washed with ethyl acetate arl(l chrolnato~Jr.lphed on a column o~ non-ionic polymer resiIl Amberlite XAD-2 (resistered trade mark, manufactured by Rohm & Haas Co., ~.S.A.), using water as an eluent. The fractions eontaining the cephalosporin compound are eollected and concentrated to dryness under reduced pressure. 0.61 g of sodium 7B-{(Z)-2-(2-aminothiazol-4-yl)-2- ~(1-methyl-2-pyrrolidon-3-~l)oxy;millo)acetamido~-3-~(1-methyl-lH-tetrazol-5-yl)thio-methyl~-3-cephem-4-carboxylate is obtained.
NMR (D2O)~: 2.1 -2.7 (2EI, m), 2.a9 (3H, s) 3.2 - 3.8 (4H, m), 4.05 (3H, s) L0 4.05 - 4.3 (2H, m), 4.9 - 5.3 (2H, m) 5.73 (lH, d, J= 5Hz), 7.00 (lH, s) Example 9 (1) 0.5 g of (Z)-2-(2-tritylaminothiazol-4-yl)-2-~(2-piperidon-3-yl)oxyimino)aeetie aeid is dissolved in 10 ml of tetrahydrofuran, and 0.47 g of benæhydryl 7~-amino-3 ((1-methyl-lH-tetrazol-5-yl)thiomethyl~-3-cephem-4-carboxylate, 0.15 g of l-hydroxybenzotriazole and 0.24 g of dieyelohexyl-earbodiimide are added thereto. The mi,cture is s-tirred at room temperature for 2.5 hours. The reaetion mixture is treate-l in the same manner as described in Exam21e 3-(1).

0.~/ g of benzhydryl 7B-~(Z)-2-(2-tritylarn:inothiazol-4-yl)-2- ~(2-p:iperidon-3-yl)o,cyimino)acetamidol-3- ~(l-methyl-l~-~etrn~l~ol-S-yl)t'-iomethyl)-3-cephem-4-carbo~cylate is obtained.
NMI~ (DMSO-d6)5: 2.00 - 2.5 (4H, m), 3.0 - 3.3 (2H, m) 3.6 - 3.3 (2~l, m), 3.a (3~1, s) 4.1 - 4.3 (2H, m), 4.6 - 4.a (lH, m)

9~

4.9 - 5 4 ~lH, m), 5.85 ~lH, d, d, J=
8Hz, J= SHz) 6.7] (lH, s), 6.81 (lH, s) 7.0 - 7.6 (26H, m), 9.63 (lH, d, J=
9Hz) (2) 0.~ g of benzhydryl 7B-~(Z)-2-(2-tritylaminothiazol-~-~1)-2-~(2-piperidon-3-yl)oxyimino)acetamido}-3-~(1-methyl-lEI-tetrazol-5-yl)thiomethyl~-3-cephem-4-carboxylate is added to a mixture of 10 ml o~ trifluoroace-tic acid and 0.8 ml of L~ anisole, and the mixture is allowed to stand at room temperature for 30 minutes. The mixture is evaporated under reduced pressure to remove trifluoroacetic acid. Ether is added to the residue, and the resulting powder is collected by filtration.
The powder is suspended in water, and sodium bicarbonate is added thereto to dissolve said powder therein. The solution is washed with ethyl acetate and chromatographed on a column of non-ionic polymer resin ~mberlite XAD-2 (resistered trade mark, manufactured by Rohm & Haas Co., U.S.A.). The column is washed with water, followed by elution with 10 % aqueous methanol. The fractions containing the cephalosporin compound are coLlected and concentrated to dryness under reduced pressure. 0.9 g of sodium 7B-~(Z)-2-(2-aminothiazol-~-yl)-2- ~(2-piperidon-3-yl)o~yimino)acetamido3-3- ~(1-methyl-lEI-ketrazol-S-yl)thiometllyl~-3-cephem-~-carboxylate is obtained.
NMR (D2O)~: 1.5 - 2.3 (~H, m), 3.0 - 3.~ (2EI, m) 3.~ - 3.6 (2EI, m), 3.90 (3~1, m) ~.0 - ~.2 (2~1, m), 5.03 (1~1, d, J- 5EIz) 5.63 (1~1, d, J= SElz), 5.85 (Lll, s) ~3~

Example 10 (1) 18.2 g of phosphorus oxychloride are added dropwise to 9.2 ml of dimethylformamide under ice-cooling, and the mixture is s-tirred at 25 to 35C for 30 minutes. After cooling to 0C 100 ml of chloroform are added to the mixure, and said rnixture is further cooled -to -35C. A solution of 20 y of (Z)-2-(2-tritylaminothiazol-4-yl)-2-((2-pyrrolidon-3-yl)-o~.yimino)acetic acid (~-isomer) and 5.6 ml of triethylamir.e in 160 ml of N, N-dimethylacetamide is added dropwise to the mi~ture at -35 to -25C, and the mixture is stirred at the same temperature for 20 minutes. Then, a solution of 7~aminocephalosporanic acid (said solution being prepared by stirriny a mixture of 16 g of 7-aminocephalosporanic acid, 48 g of trimethyl chlorosilane, j35.6 ml of pyridine and 160 ml of N, N-dimethylacetamide at 10 to 20C for 2 hours) is added dropwise to the reaction mixture at -35 t`o -20C
under stirring. After s-tirring at the same -tem~erature for 20 minu-tes, said reaction mi~ture is poured into 2 liters of ice-water and s-tirred vigorously. Crystalline precipitates are collected by filtration, washed with water, ethyl acetate and ether, and then dried in vacuo. 26.5 g of 7B-~(Z)-2-(2-tritylaminothia~ol-4-yl)-2-~(2-pyrrolidon-3-yl)o~ ino)acetamido}-cepllalosporanic acid (~-isomer) are obtained as colo~-less powder.

~R (DMS0-d6)~ : 2.06 (3H,s), 2.1 - 2,5 (2H, m), 3.1 - 3.4 (2H, m), 3.45 (lH, d, J=
18~Iz), 3,75 (lH, d, J- 18Hæ) 4.6 - 5.02 (3~I, m), 5.17 (lH, d, J= 5.~1z), 5.6 - 5.9 (lH, m), 6.81 (lH, s), 7.1 - 7.6 (15H, m), 7.97 (lH, s), 8.88 (lEI, br, s), 9.64 (lH, d, J=6Hz) (2) 67.5 g of sodium iodide and 10.9 g of pyridine are acided to a mixture o~ 18 ml of water and 18 ml of dimethyl-L~ ~ormamide, and the mixture is heated to 80C. 11.5 g of 7~-¦(Z)-2-(2-tritylaminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)-oxyimino)acetamido}cephalosporanic acid(~-isomer) are added to the mixture. The mixture is stirred at 80C for 35 rninutes. After cooling, 90 ml of ice-water are added to the reaction mixture, and said mixture is washed with ethyl acetate. Then, the reaction mixture is evaporated under reduced pressure to remove ethyl acetate and adjusted to pH
2 with 10 6 hydrochloric acid. Crystalline precipitates are collected by filtration, washed with water and dried in vacuo. 11.9 g of 73-~(Z)-2-(2-tritylaminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oxyimlno)acetamidol-3-(1-pyricliIliomethyl)-3-cel?ile~m-~l-carhoxylate (,e-isomer) are obtained as crucl~ pale yellow powc1er.
(3) 5.5 g of 7B-~(Z)-2-(2-tritylamino-thiazol~l-yl)-2- ~(2-py:rro:l;.don--3-yl)oxyimino~acetalnido¦-3-(1-pyri.diniomethyl)-3-cephem-~ carboxylate (Q-;.somer) obtained above are dissolved in 90 ~ ~3~

ml of 80 % formic acid. The solution is stirred at room temperature for one hour. The reaetion solution is coneentrated under redueed pressure, and 200 ml of water are added to the rrs:Lclue. Insoluble Materials are filtered off. The ;~C3~1e-~OUS Eiltrate is washed ~ith ethyl ae2tate, eoncentrated under rr-~clucr_d pressure to remove ethyl aeetate ancl then introduced into a eolumn paeked with 200 ml of a non-ionie a~sorption resin (manufaetured by Mitsubishi Chemieal Industries Ltd. under the trade name "Diaion HP-20"). After the eolum :L~ is washed wit'n water, said eolumn is eluted with 20 ~ aqueous rnethanol. The eluate is eoncentrated under reduced pressure, aeetone is added to the residue and the resultant precipitates are colleeted by filtration. 1.6 g of 7~-~(Z)-2-(2-aminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oxyimino)acetamido}-3-(1-pyridinio-methyl)-3-eephem-4-earboxylate (~-isomer) are obtained as pale ~ellow powder.
The physicochemieal properties of this product are identieal with those oE the sample obtained in E~ample 6.
Rxample lL

~ _ .
2.5 g oE 7~3-¦(Z)-2-(2-tritylamino-thiazol~~-~l)-2-~(2-p~rrolidorl-3-yl)ox~iml.no)aeetamido3cephalosporarlic acid (~~isomcr) obtclinecl in the samc manner as dr-~scrihed in ~.~alllplre 10 - (L~ arc dissolvcd in 50 ml of ~0 ~ formic acir~, arl-l the sol~ltion is c;tirred at room tempertlturi- for one hour. InsoLuble materials are filtered off. Thc filtrate i; conccntrated ullder rcduc~d prcss~lr-, ~at~r is allcled to ~3~

the residue, and the aqueous mixture is neutralized with sodium bicarbonate. Then, said aqueous mixture is washed with ethyl acetate, concentrated under reduced pressure and introduced into a column packed with 200 ml of a non-ionic aGIsorption resin ("Diaion HP-20"). After the colurnn is washed ~lith ~ater, said column is treated in the same manner as dc.scribcd in Example 10 - (3). 1,0 g of sodium 7B-~(Z)-2-(2-alninothiazol--4-yl)-2- ~(2-pyrrolidon-3-yl)oxyimino3acetamido3-cephalosporanate (~-isomer) is obtained as pale yellow L~ powder. Another designation of this levorotatory isomer is shown as sodium 7B-~(Z)-2-(2-a~inothiazol-4-yl)-2--(((3S)-2-pyrrolidon-3-yl)-oxyimino3acetamido}cephalosporanate.
NMR (D2O)~ : 2.12 (3H,s), 2.3 - 2.6 (2H, m), 3.2 - 3.9 (4H, m), 4.75 (lH, d, J= 13Hz), 4.95 (lH, d, J= 13Hz), 5.10 (lH, t, J= 7Hz), 5.25 (lH, d, J= 5Hz), 5.85 (lH, d, J=5Hz), 7.08 (lH, s) The product obtained above is treated in the same IllannCr c-lS described in Exarnpl-~ 7, whereby 7B-¦(Z)-2-(2-amino--2() thi.a~ol-LI-yI)-2-~(2-pyrrolidon-3-~fl)oxyimi.no)acetamido3-3-(1-p~ridi.n:iormcthyl)-3-cephen-L1-cclrboxylate (,e-i.somer) is obl:lli.n(!d.
t2 /~ nli.xui re oE 960 nly oE 7~ (Z)-2-(2-aillirlot~i.aæol-LI--yl)-2-~( 2-p~frr.o:L .idoll-3-y 1 )o,~.i.millo~acctclllli(lo}ccphalospo~ nlc ~:3~

-39- , acid, 730 mg of isonicotinamide, 4.5 g of sodium iodide and 3 ml of water is stirred at 65 to 70C for 6 hours. After cooling 20 ml of water are added to the reaction mixture, and lnsoluble materials are filtered off. The filtrate is :introduced into a column packed with 100 ml of a non-ionic cldsorpkion resin ("Diaion HP-20"). The column is washed with wllter. Then, said column is eluted with 20 ~ aqueolls m~khanol, and the eluate is concentrated under reduced pressure. 175 mg of 7B-~(Z)-2-(2-aminothiazol-4~yl)-2-~(2-pyrrolidon-3-yl)-oxyimino~acetamido}-3-(4-carbamoyl-1-pyridinio-rnethyl)-3-cephem-A-carboxylate are obtained as pale yellow powder.
m.p. 163 - 166C (decomp.) NMR (CF3CO2D) ~ : 2.5 - 3.0~ (2H~ m), 3.5 - 4.0 (4H, m), 5.2 - 5.5 (3H, m), 5.5 - 5.8 (lH, m), 5.9 - 6.2 (lH, m), 7.35 (lH, s), 8.4 - 8.7 (2H, m~, 9.1 - 9.4 (2H, m) xample 13_ A mi~cture of 960 m~ of 7B-~(Z)-2-(2-aminothiazol-4-yl)-2-~(2 ~o pyrrolidon-3-yl)oxyimino~ac~tamido3cephalosporanic acid, 650 rn~] o~ ~-hydroxym~thylpyridine, ~.5 ~ oE sodium ;odide and 3 ml Oe water is stirred at 65 to 70C Eor 7 hours. Ttl~
ttle re~lckiotl mi:cture is treated in th~ same mallner as described irl l.x.lmple 12. 150 m~ of 7B-¦(Z)-2-(2-aminoth:ia~ol-4-yl) 2-~(2-~yrroLldon-3-yl)oxyilll:ino~-acetamido~-3-(~l-hydro.cym~thyl-1-pyri;liniomethyl)-3-cep~lem-~-carboxylate ar~? ohtaineci as pal2 yelLow powd~r.

m.p. 162 - 184C (decomp.) NMR (D2O) ~ : 2.3 - 2.7 (2H, m), 3.1 - 3.7 (4H, m), 4.8 - 5.0 (2H, m), 5.03 (2H, s), S.24 (lH, d, J= SHz), 5.3 - 5.6 (lH, m), 5.83 (lH, d, J= 5Hz), 6.81 (lH, s), 7.8 - 8.2 (2H, m), 8.6 - 9.0 (2H, m) E:x;,lrn,e,~ e 14 A mixture of g60 mg of 7B-¦(Z)-2-(2-aminothiazol-4~yl)-2-~(2-pyrrolidon-3-yl)oxyimino)acetamido}cephalosporanic acid, I0 650 rng of 3-hydroxymethylpyridine, 4.5 g of sodium iodide and 3 ml of water is treated in the same manner as described in Example 12. 83 mg of 7B-~(Z)-2-(2-aminothiazol-4-yl)-2- ~(2-pyrrolidon-3-yl)oxyimino)-acetamido~3-(3-hydro~ymeth pyridiniomethyl)-3-cephem-4-carboxylate are obtained.
m.p. 128 - 135C (decornp.) NMR. (D2O) ~ : 2.1 - 2.8 (2H, rn), 3.1 - 3-8 (~H, m), 4.86 (2H, s), 4.9 - 5.1 (2H, rn), 5.27 (lH, d, J= 5rIz) ~ 5. 3 - 5.6 (lFI, m), 5.83 (lH, d, J= 5Hz), 6.96 (lH, s), 7.7 - 8.2 (1~l, m), 8.3 - 8.6 (1~-l, m), 8.6 - 9.1 (2H, m) Ur~ tion o~ Startir~ co~ounds:
(1) L5.8 cJ o~ rethy~ 2-(2-tritylarninott-l:icn~ol-~ 2-hydro~Yyilnirloac~tate are dissolvrcl in 70 ml o~ dim~thylsul oxic~e, a~l~l 5.8 cJ o~ anhydrous potassium carbonate ar~ ackle-l th~reto.

~.~3~

The mixture is stirred at room temperature for 20 minutes.
6.6 g of 3-bromo-2-pyrrolidone are added to said rnixture, and the rni~ture is sti.rred at room temperature for 20 hours.
Th~ mi~-ture is poured into 800 ml of water, and crys-talline pr~ci.pitatfs are collected by filtration and washed wi-th water. The cr~stals are dissol~ed in chloroform, washed with water and then dried. Then, the chloroform solution is cvaporatcd under red~lced pressure to remove solvent.
100 ml oE ethyl acetate are added to the residue, and allowed :L() to stand at room temperature. Crys-talline precipitates tnus obtained are collected by filtration and dried. 16.0 y of ethyl (Z)-2-(2-tritylaminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oxyimino)ace-tate are obtained.
M.p. 209 - 210C
~R (CDCl3)~: 1.30 (3H, t, J= 7Hz), 2.1 - 2.6 (2H, m) 3.1 - 3.6 (2H, m), 4.34 (2E~,`q, J= 7Hz) 4.90 (lH, t, J= 7Hz), 6.53 (lH, s) 7.0 - 7.6 (17H, m) 16.0 g of ethyl (Z)-2-(2-tritylaminothiazol-4-yl)-2- ~(2-p~rroli.don-3-yl)o~yimino~acetate are aclded to a rni~ture o~
160 ml of methanol and 30 ml of an aqut-~ous 2N sodium hydro~:ide SOlllt.iOIl, allCI the mi~ture is re~flu~ed for 30 mi.nuteC; undt-~r ileatin~J. ~Eter cooling, crystalline prcc~ itat~s are collccted by ~iltration and washed with rnethanol. The crystclls are susp~llded in 30 ml of wat:er. Then, the~ suspellsio .;.s ~djuste(-l to p~l 3 wi.th 2N hydrochloric acid. CrystaLline ~23~

precipitates are collected by filtration and dried. ll.4 g of (Z)-2-(2-tritylaminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)-oxyirnino)ace-tic acid are obtained.
M.p. 150 - 153C (decomp.) NM~ (DMSO-d6)~: 1.8 - 2.4 ~2H, m), 2.9 - 3.4 (2H, m) .63 (lH, t, J= 7Hz), 6.76 (lH, s) 6.9 - 7.6 (lSR, m), 7.85 (lH, s) 8.70 (lH, broad 5) (2) 30 cJ of (Z)-2-(2-tritylaminothiazol-4--yl)-2- ~(2-L0 pyrrolidon-3-yl)oxyimino)acetic acid and 60 ml of methanol are added to 100 ml of dioxane containing lO.5 g of methyl L-phenylalaninate, and t'ne mixture is heated a-t 50C to dissol~e said acid therein. 700 ml oE dioxane are added to the solution, and the mixture is stirred at room temperature for 5 hours. Crystalline precipitates are collected by filtration (the ~iltrate is hereinafter referred to as "E'iltrate I"), and 14.3 g of t~e crude product thus obtained are dissolved in 24 ml of methanol. 280 rnl of dioxane are added to the methanol solution. The mixture is stirred at roorn terllperature for ~I hours, and crystalline precipitates are collected by Eiltration (th~ filtrate is hereirlafter re~f(!trecl to as "F'iltrate [I"). 12.2 9 of ('b)-2-(2-t-rityl-allli.rlot^hi<-l~c)1-~1-y])-2-((2-pyrro1iclorl-3-yl)oxyimiIlo)acetic cid (~-isomer) methyL L-phenylalaninate salt are obtainec1.
~)25 _ l~I.0 (C'= l, methc~nol) 12.2 ~J oE thc above-mentioned salt are dissolved in 120 9~a3 ml of methanol, and 176 ml of O.lN hydrochloric acid are added thereto. The mi~ture is stirred Eor 2 hours under ice-cooling. Crys-talline precipitates are collected by filtration and ~ashed with methanol. 7.5 g of (Z)-2-(2--tritylaminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oxyimino3acetic acid (~-isorner) are obtained. Another designation of this Lc!vorot,ltc)ry isomer is sho~n as (Z)-2-(2-tritylaminothiazol-4-~yL)--2-~((3S)-2-pyrrolidon-3-yl)oxyimino)acetic acid.
M.p. 1~2 - 143C(decomp.) )25 _ 38.8 (C= 1, dimethylformamide) (3) Filtrate I & II obtained in the above men-tioned parayraph (2) are condensed to dryness under reduced pressure.
The residue is dissolved in 250 ml of methanol and then 450 ml of 0.1 N hydrochloric acid are added dropwise to the solution. The mi~ture is stirred for 2 hours under ice-cooling.
l'he resulting crystalline precipitates are collècted by filtration, washed with methanol, and dried. 20 g of (Z)-2-(2-tritylaminothiazol-4-yl)-2-~(2-pyrrolidon-3-yl)oA~yimino~-acetic acid (containing e~cess of the d-isomer) are obtained.
20.0 y of (Z)-2-(2-tritylamirlothiazol-~-yl)-2-~(2-pyrrolidon-3-yl)oxyimino)acet:ic acid th~ls recovered and ~10 ml of methanol are a(lded to 7() ml oE dioxane containiny 7.0 y of methyl l)-ph(~llyLtl]anillate~ arlcl the mixture i5 heated at 50C to di~;c;olve r~ i(l acid therci~ 50 ml of dio:;ane are aclde(l to s.lid ~-;olution. Til~!rl, the mixture is stirrc~cl at room telllpor-akul:e Eor ~I hours, and crystalline precipitato~ aro collected by filtration. 13.3 g of the erude product thus obtained are dissolved in 20 ml of methanol, and 260 ml of dio~ane are added thereto. The mixture is stirred at room temperature for 4 hours. Crystalline preeipitates are collected by Lltration. 12.0 g of (Z)--2-(2--tritylaminothiazol-4-yl)-2-~ 2-pyrrolidon-3-yl)oxyirnino)aeetic acid (d-isomer) methyl l)-ph~Jnylal,lninate salt are obtained.
2 ~ 13.9~ (C= 1, methanol) 'L2.0 g oE the above-mentioned salt are dissolved in 120 ],~ tlll o methanol, and 174 ml of 0.1 N hydrochlorie acid are acld~d thereto. The mixture is stirred for 2 hours under iee~eooliny. Crystalline precipitates are eollec-ted by fi'ltration and washed with methanol. 7~3 g of (Z)-2-(2-trityl-aminothiazol-4-yl)-2- ((2-pyrrolidon-3-yl)oxyimino~acetie aeid (d-isomer) are o~tained. Another designation of this de~trorotatory isomer is shown as (Z)-2-(2-tritylaminothiazol-~-yl)-2-~((3R)-2-pyrrolidon-3-yl)oxyimino)aee-tic aeid.
M.p, 143 - 144C (deeomp.) ~25 ~ 37,4O (C= 1, dirnethylformarnide) ~o (~) 2,7 y of ethyl (Z)-2-(2-tritylaminothiazol-4-yl)-2-hyclroxyilnlnoaeetate arc c~issolved in 12 ml of dimethylsuloxide, alld t.0 cJ oF arlhydrous potclssium earbonate is addecl thereto ullcl{,~r nLtrO~Jen gas atmosE)Iler~. The mi~cture is stirred at roolrl teln2erc~ture for 10 rninutes. 1.2 g oE l-rnethyl-3-bron)o-2-pyrroLiclorle are added to the mixture, ancl ~he mixture is stirrcd at room terlloerature Eor 5 hours, rrhe reaction ~3~

mixture is poured in-to lO0 ml of water, and crystalline Drecipitates are collected by filtration. The crystals are dissolved in ethyl acetate, and the solution is washed with water and then dried. The solution is concen-trated under reduced pressure to remove solvent. Then, the residue is crysta]li7ed with isopropyl ether ancl collected by filtration.
2.1 y o~ ethyl (~)-2-(2-tritylaminothiazol-4-yl)-2-~(l-methyl-2-~)yrr:olidon-3-yl)oxyimino)acetate are obtained.
N~P~ (CDCl3)~: 1.30 (3M, t, J= 7Mz), 2.0 - 2t7 (2H, m) :L~ 2.88 (3FI, s), 3.0 - 3.6 (2H, m) 4.34 (ZH, q, J= 7Hz), 4.92 (lH, t, J-7Hz), 6.54 (lH, s), 6.87 (lH, s) 7.0 - 7.5 (15H, m) 2.7 g of ethyl (Z)-2-(2-tritylaminothiazol-4-yl)-2-~(l-me-thyl-2-pyrrolidon-3-yl)oxyimino)acetate are suspended in 27 ml,of methanol, and 4.9 ml of 2N sodium hydrc~xide solution are added thereto. The mixture is refluxed for 20 minutes under heating. Alter cooling, the mixture is concentrated under reduced pressure to remove methanol. The residue is adjusted to pFI 3 with 2N hydrochloric acid and extracted ith ethyl acetate. The extract is dried and evaporated un(ler reclucecI pr-.ssure to r-move solvent. Therl, the resicllle tnus obtairled is crystalli~ed .~ith ekher and collecttd by eiltratiotl. 2.15 g of (Z)-2-(2-tritylaln;rlothi.l7Ol-~l-yl)-2-~I-mcthyl-2-pyrrolicloIl~3-yl)oxyimi~ acet;c ac~;.d are ohtaiIled.

M.p. 142 - 145C (decomp.) NMR (DMSO-d6)r: 2.0 - 2.5 (2H, m~, 2.77 (3H, s) 3.1 - 3.4 (2H, m), 4.78 (lH, t, J= 8Hz) 6.87 (lH, s), 6.9 - 7.5 (16H, m) (5) 1.3 g of (Z)-2-(2-tri~ylaminothia~ol-~-yl)-2-hydroxy-irninoacetlc acid are dissolved in 10 ml of dimethylformamide, and 0.2~ y of sodium hydride (60 ~ oil dispersion) is added thereto. The mixture is s-tirred at room -temperature for 15 mirlute~C.. 0.65 g o~ 3-bromo-2-piperidone is added to the L0 rnixture, ancl the mi:cture is s-tirred at room temperature for 1.5 hours. The reaction mixture is poured into water and ~"ashed with a mixture of ethyl acetate and tetrahydrofuran (1 : 1). The aqueous layer is adjusted to pH 3 with 10 %
hydrochloric acid and extracted with a mixture of ethyl acetate and tetrahydrofuran (1 : 1). The extract is dried and concentrated to dryness under reduced pressure. Then, ether is added to -the residue,-and the resulting powder is co]lected by filtration. The powder (1.3 g) is purified by silica gel chromatoyraphy (solvent, methanol: chloroform =
1: ~)- 0.85 g of (Z)-2-(2-tritylaminothiazol-4-yl)-2-((2-pipericlon-3-yl)oxyimino)acetic acid are obtained.
M.p. 1~15 - 150C (decomp.)

Claims (12)

Claims:

1. A process for preparing a 7 .beta.-[(Z)-2-(2-aminothiazol-4-yl)-2-oxyimino-acetamido]-3-cephem-4-carboxylic acid compound of the formula:

(I) wherein R1 is hydrogen or lower alkyl;
R2 is acetoxy or (1-methyl-1H-tetrazol-5-yl)thio, and R3 is carboxy; or R2 is a group of the formula: , R3 is -COO- and Y is hydrogen, hydroxymethyl or carbamoyl; and n is an integer of 2 or 3, or a pharmaceutically acceptable salt thereof; which process either comprises (A) the steps of:
(i) condensing a compound of the formula:

(II) wherein R4 is a protecting group and R1 and n are the same as defined above, or a reactive derivative thereof with a compound of the formula:

(III) wherein R5 is a carboxy or a protected carboxy when R2 is acetoxy or (1-methyl-1H-tetrazol-5-yl)thio: or R5 is -COO- when R2 is a group of the formula: , and Y is the same as defined above, or a salt thereof to give a compound of the formula:
(IV) wherein R1, R2, R4, R5 and n are the same as defined above, and (ii) removing the protecting group or groups therefrom, and (iii) if required, further converting the product into a pharmaceutically acceptable salt thereof; or for preparing a 7.beta.-[(Z)-2-(2-aminothiazol-4-yl)-2-oxyimino-acetamido]-3-cephem-4-carboxylic acid compound of the formula:
(I-a) wherein R1 is hydrogen or lower alkyl;
R6 is (1-methyl-1H-tetrazol-5-yl)thio, and R3 is carboxy; or R6 is a group of the formula: , R3 is -COO- and Y is hydrogen, hydroxymethyl or carbamoyl; and n is an integer of 2 or 3, or a pharmaceutically acceptable salt thereof, (B) the steps of:
(i) reacting a compound of the formula:

(V) wherein R7 is hydrogen or a protecting group and R1 and n are the same as defined above, or a salt thereof with a pyridine compound of the formula: (wherein Y is the same as defined above), (1-methyl-5-mercapto)tetrazol or a salt thereof, (ii) when R7 is a protecting group, removing same protecting group therefrom, and (iii) if required, further converting the product into a pharmaceutically acceptable salt thereof.

2. A process according to claim 1 for preparing a compound of formula (I) in which R1 is hydrogen or methyl; R2 is acetoxy or (1-methyl-1H-tetrazol-5-yl)thio, and R3 is carboxy; or R2 is a group of the formula: , and R3 is -COO- and Y is hydrogen, hydroxymethyl or carbamoyl; and n is an integer of 2 or 3; which process comprises carrying out steps (A) employing starting materials (II) and (III) in which R1, R2 and n are as defined above and R4 and R5 are as defined in claim 1.

3. A process according to Claim 1 for preparing a com-pound of formula (I) or (Ia) in which R1 is hydrogen or methyl; R2 is (1-methyl-1H-tetrazol-5-yl)thio, and R3 is carboxy; or R2 is a group of the formula: R3 is -COO- and Y is hydrogen, 4-hydroxymethyl, 3-hydroxymethyl or 4-carbamoyl; and n is an integer of 2 or 3; which process comprises carrying out steps (A) using starting materials (II) and (III) in which R1, R2 and n are as defined above and R4 and R5 are as defined in claim 1;

or carrying out steps (B) using a starting material (V) in which R1 and n are as defined above and R7 is as defined in claim 1.

4. A process according to Claim 1 for preparing a com-pound of formula (I) in which R1 is hydrogen; R2 is a group of the formula: , R3 is -COO-, and Y is hydrogen, 4-hydroxymethyl, 3-hydroxymethyl or 4-carbamoyl;
and n is an integer of 2; which process comprises carrying out steps (A) using starting materials (II) and (III) in which R1, R2 and n are as defined above and R4 and R5 are as defined in claim 1; or carrying out steps (B) using starting material (V) in which R1 and n are as defined above and R7 is as defined in claim 1.

5. A process according to Claim 1 for preparing 7.beta.-{(Z)-2-(2-aminothiazol-4-yl)-2-[(2-pyrrolidon-3-yl) oxyimino]acetamido}-3-(1-pyridiniomethyl)-3-cephem-4-carboxy late or a pharmaceutically acceptable salt thereof;
which process comprises carrying out steps (A) using starting materials (II) and (III) in which R1 is hydrogen; R2 is a group of the formula: , R4 is a protecting group, R5 is -COO- and n is 3; or carrying out steps (B) using a starting material (V) in which R1 is hydrogen, R7 is hydrogen, or a protecting group, and n is 3 and reacting said compound with pyridine; and if necessary reacting the product with a pharmaceutically acceptable base.

6. A process according to Claim 1 for preparing 7.beta.-{(Z)-2-(2-aminothiazol-4-yl)-2-[((3S)-2-pyrrolidon-3-yl) oxyimino]acetamido}-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate or a pharmaceutically acceptable salt thereof;
which process comprises carrying out steps (A) using start-ing materials (II) and (III) in which R1 is hydrogen;
R2 is a group of the formula: R is a protecting group, R5 is -COO- and n is 3; or carrying out steps (B) using a starting material (V) in which R1 is hydrogen, R2 is hydrogen or a protecting group and n is 3, and reacting said compound with pyridine; wherein steps (A) or steps (B) or either carried out using a starting material suitable to provide the levorotatory product, or a racemic starting material is used and the levorotatory product is separated from the resulting racemic product mixture; and if neces-sary reacting the product with a pharmaceutically accept-able base.

7. A 7.beta.-[(Z)-2-(2-aminothiazol-4-yl)-2-oxyimino-acet-amido]-3-cephem-4-carboxylic acid compound of the formula:

(I) wherein R1 is hydrogen or lower alkyl;
R2 is acetoxy or (1-methyl-1H-tetrazol-5-yl) thio, and R3 is carboxy; or R2 is a group of the formula: R3 is -COO- and Y is hydrogen, hydroxymethyl or carbamoyl; and n is an integer of 2 or 3, or a pharmaceutically acceptable salt thereof; whenever prepared by the process of Claim 1 or an obvious chemical equivalent thereof.

8. A compound according to Claim 7, in which R1 is hydrogen or methyl; R2 is acetoxy or (1-methyl-1H-tetrazol-5-yl)thio, and R3 is carboxy; or R2 is a group of the formula: , and R3 is -COO- and Y is hydrogen, hydroxymethyl or carbamoyl; and n is an integer of 2 or 3; whenever prepared by the process of Claim 2 or an obvious chemical equivalent.

9. A compound according to Claim 7, in which R1 is hydrogen or methyl; R2 is (1-methyl-1H-tetrazol-5 yl) thio, and R3 is carhoxy; or R2 is a group of the formula: , R3 is -COO- and Y is hydrogen, 4-hydroxy-methyl, 3- hydroxymethyl or 4-carbamoyl; and n is an integer of 2 or 3; whenever prepared by the process of Claim 3 or an obvious chemical equivalent.

10. A compound according to Claim 7, in which R1 is hydrogen; R2 is a group of the formula: , R3 is -COO- and Y is hydrogen, 4-hydroxymethyl, 3-hydroxymethyl or 4-carbamoyl; and n is an integer of 2; whenever prepared by the process of Claim 4 or an obvious chemical equivalent.

11. A compound according to Claim 7, which is 7.beta.-[(Z)-2 (2-aminothiazol-4-yl)-2-[(2-pyrrolidon-3-yl) oxyimino]
acetamido}-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate or a pharmaceutically acceptable salt thereof; whenever prepared by the process of Claim 5 or an obvious chemical equivalent.

12. A compound according to Claim 7 whieh is 7.beta.-{(Z)-2-(2-aminothiazol-4-yl)-2-[((3S)-2-pyrrolidon-3-yl) oxyimino]
acetamido}-3-(1-pyridiniomethyl)-3-cephem-4-carboxylate or a pharmaceutically acceptable salt thereof; whenever prepared by the process of Claim 6 or an obvious chemical equivalent.