CA1042422A - 7-(substituted phenylglycinamido)-3-substituted-3-cephem-4-carboxylic acid derivatives and preparation thereof - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE

Compounds of the formula:

Description

~o~ z~
This inventlon relates to new cephalosporarlic acid derivatives which possess an antibacterial activity, processes for preparing the same and a composition thereof.
Cephalosporanic acid derivatives are described which can be represented by the following general formula:

COOH

wherein Rl is nitro group, alkoxy group, alkanesulfonamido group, alkylaminosulfonamido group, alkylureido group or alkylthioureido group, R2 is hydrogen atom or hydroxy group, provided that R2 is hydroxy group when Rl is nitro group or alkoxy gxoup. R3 is hydrogen atom or heterocyclic-thio group, in which the heteroeyclie group may be substituted with alkyl group, provided thak R3 is hydrogen atom when Rl - ~
is alkoxy group and R2 is hydroxy group; X is amino group ~ -or proteeted amino group and nontoxie, pharmaeeutically aeceptable salts and esters thereof.
The term "alkoxy group" represented by Rl suitably ineludes compounds having 1 to 9, preferably 1 to 6 carbon B~

. .
.

4~Z
atom~, for example, methoxy, ethoxy, propoxy, i30propoxy, butoxy, i30butoxy, tert-butoxy, pentyloxy, hexyloxy, heptyl-oxy, oct~loxy, nonyloxy, and the like.
The term "alkane~ulfonamido group" represented by Rl mean~ mesylamino, ethanesulfonamido, propanesulfonamido, i~opropane~ulfonamido, butanesul-fonamido, i90butane~ulfonamido, tert-butanesulfonamido, pentanesulfonamido, hexanesulfonamido, and the like.
The term "alkylaminosulfonylamlno group" repre-sented by Rl mean~ alkyl 8ubstituted amino~ulfonylamino - group, in which the alkyl group has the same meaning as the alkyl moiety in the "alkanesulfonamido group"
mentioned abQve.
The term "alkylureido group" represented by R
m~an~ alkyl substituted ureido group, in which the alkyl group ~a~ the ~ame meaning as the alkyl moiety in the "alkanesulfonamido group" mentioned above The term "alkylthioureido group" represented by Rl means alkyl sub~tituted thioureido group, in which the alkyl group ha~ the same meaning as the alkyl mDiety in the "alkane~ulfonamid~ group" mentioned above.
The term "heterocyclic-thio group" represented by R3 means a residue of a thiol compound having a heterocyclic gxoup, in which the heterocyclic group mean3 a re~idue of furan, thiophene, pyrrole, pyrazole, imidazole, triazole, thiazole, i~othiazole, oxazole, isoxazole, thiadiazole, oxadiazole, thiatriazole, .
.. . . ..
: , ,'' , 'i' :',. ' ': ' ' ~CI 4~9~2Z
oxatriazole, tetrclzoLe, pyridine, pyrazine, pyrimidine, pyrid-azine, benzothiop~erle, bonzofuran, inclole, in~azole, benzimid-azole, benzothiazol, benzothiadiazole, benzoxazole, purine, quinoline, isoquinoline, phthalazine, naphthyridine, quinoxa-line, quinazoline, pyrrolidine, imidazolidine, piperidine, piperazine, or the like. The heterocyclic moiety of the "heterocyclic-thio group" may be substituted with one or more alkyl groups, e.g., rnethyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, and the like.
The term "protected arnino group" represented by X
means an amino group protected by the conventional amino pro-tecting group, e.g., benzyloxycarbonyl, substituted benzyloxy-carbonyl, alkoxycarbonyl, substituted alkoxycarbonyl, aralkoxy-carbonyl, adamantyloxycarbonyl, trityl, substituted phenylthio, substituted aralkylidene, substituted alkylidene, substituted cycloalkylidene and the like.
The term "nontoxic, pharmaceutically acceptable salt"
means a salt of an alkali metal cation e.g., sodium, potassium and the like.
Tha alkyl and alkoxy portions of the various groups represented by ~1 and R3 suitably contain from 1 to 9 carbon atoms and preferably from 1 to 6 carbon atoms.
The present invention is concerned with new compounds within the aforementioned formula, of a general formula (I) ~' H-CO~

COOH

wherein Rl is nitro or a lower alkoxy group, and nontoxic, pharmaceutically acceptable salts and esters thereof.

_ 3 - . , . ~ ~ , .

~LO~
The clescrihed class of cephalosporanic acid derivative3 may be p.repared by reacting 7-amino~3-substituted-3~cephem-4-carboxylic acicls o~ .Eormul.a:

H2~ r--~ ~
/ ~ N \ ~,~ CH~_R3 (II) OOH
wherei.n R3 is as defined above, or derivatives at the amino group and/or the carboxy group, with substituted phenylglycine of formula:

~ ~H - COOH (III) wherein Rl, R2 and X are as defined above, or their reactive derivatives at the carboxy group.
According to the present invention there is provided a process for preparing the new compounds of formula (I) by the above identified acylation reaction, wherein R3 in said compound of formula (II) is hydrogen, and, in the substituted phenylglycine of formula (III), R2 is hydroxyl~
The derivative at the carboxy group of the compound (II) may be a salt e.g., magnesium salt, calcium salt, triethylamine salt, etc.' an ester e.g., methyl ester, ethyl ester, propyl ester, butyl ester, pentyl ester, trimethyl-silyl ester, 2-mesylethyl ester, 2-iodethyl ester, 2,2,2-tri-. .

~o~z~
chl.oroethyl est~r, benzyl ester, ~~methoxyberl~yl ester, 4-nitrobenzyl. es-ter, phenacyl ester, phenethyl ester, trityl ester, diphenylmethyl ester, bis(methoxyphenyl)methyl ester, 3,4-dimethoxybenzyl ester, (l-cyclopropyl)ethyl ester, ethynyl ester, 4-hydroxy-3, 5-di-tert-butylbenzyl ester, etc., an activated amide, an acid anhydride, an acid halide, or the like.
The derivative at the amino group of the compound (II) may be the reaction product of the - 4a -~P

~4~2~:
compound(II) and a silyl compound such as bis(trimethylsilyl)acetamido, or the like.

The reactive deriva-tive at the carboxy group of the substituted phenylglycine (III) may be an acid halide, an acid anhydride, an activated amide, an acti~ated ester, or the like. The sui-table examples may be an acid chloride, an acid azide;
a mixed acid anhydride with an acid, for example, dialkylphosphoric acid, phenylphosphoric acid, diphenylphosphoric acid, dibenzylphosphoric acid, halogenated phosphoric acid, dialkylphosphoric acid, sulfurous acid, thiosulfuric acid, sulfuric acid, alkylcarbonic acid, aliphatic carboxylic acid ~ ~
(e.g. pivalic acid, pentanoic acid~ isopentanoic ~ :
acid, 2-ethylbutyric acid or trichloroacetic acid) or aromatic carboxylic acid (e.g. benzoic acid), or a symmetrical acid anhydride; an acid amide with imidazole 9 4-substituted imidazole, dimethylpyrazole, triazole or tetrazole; or an ester (e.g. cyanomethyl ester, methoxymethyl ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, pentachlorophenyl ester, methanesul~onylphenyl ester, phenylazophenyl ester, phenyl thioester, p-nitrophenyl thioester, p-cresyl thioester, car~oxymethyl thioester, pyranyl ester, pyridyl ester, piperidyl ester, 8~quinolyl thioester, or an ester with N, N-dimethylhydroxylamine, l-hydroxy-

2-~lH)-pyridone, N-hydroxysuccinimide or N-hydroxyphthalimide), or the like. The suitable 3o .. ..

~04Z4~2Z
derivative can be optionally selected from them accor-ding to the kincl of the ~ubstituted phenylglycine (III~ to be used practically.
The reaction i~ u~ually carried out in a solvent, for example, acetone, dioxan, acetonitrile, chloro~orm, methylene chloride, ethylene chloride, tetrahydrofuran, ethyl acetate, dimethylformamude, pyridine or any other organic Aolvent inert to the reaction. Among these solventq, hydrophilic solvents may be uqed in a mixture with water.
~en the substituted phenylglycine (III) is used in a form of he free acid or its calt ln thiA
reaction, the reaction i~ preferably carried out in the pre~ence of a conden~ing agent, for example, N,N'-dicyclohexylcarb~diimide, N-cyclohexyl-N~-morpholino-ethylcarbodiimide, ~-cyclohexyl-N'-(4-diethylamino-cyclohexyl)carbodiim1de, ~,~'-diethylcarbodiimide, diisopropylcarbodiimide, ~-ethyl-~'-(3-dimethyl-aminopropyl)carb~diimide,~M,~'-~arbonyldi(2-m~thyl-imidazole), pentamethyleneketene-N-cyclohexylimine, diphenylketene-N-cyclohexylimine, alkoxyacetylene, l-alkoxy-l-chloroethylene, trialkyl phosphite, ethyl polyphosphate, isopropyl polyphosphate, phosphorus oxychloride, phosphorus trichloride~ thionyl chloride~ oxalyl chloride, triphenylphosphine, 2-ethyl-7-hydroxyben2isoxazolium salt, 2-ethyl-5-(m-sul~ophenyl)isoxazolium hydroxide intramolecular salt, (chloromethylene)dimethylammonium chloride, .

4~Z
and the like, The ~alt of the ~ub~tituted phenylglycine (III~ may be an alkali metal salt, an alkaline earth metal salt, an ammonium salt, a salt with an organic base such a~ trimethylamine, dicyclohexyl-amine or the like.
The reaction may be carried out in the presence of a base e.g., alkali metal bicarbonate, tri-alkylamine, ~,N-dialkylbenzylamine, pyridine, a~d the like. When the base or the condensing agent is in li~uid, it can be used also as a solvent. ~he reaction temperature is not restrictive, and the reaction i~ usually carried out under cooling or at room temperature.
Some of the amQno protecting groups of the protected amino group~ represented by the symbol X
in the substituted phenylglycine (IIIj may be removed in the course of the reaction or the post-treatment to give directly the objective compounds (I) having a free amino group for X.
When the reaction product has the protected amino - group, th~ amino protecting group may be removed from the reaction product, when desired, by applying a suitable removing reaction as mentioned below.
The remo~ing reaction of the amino protecting group may be carried out by a conventional method ~uch as decomposition by acid, catalytic reduction, : . : -, ,, ~ , . .

-4~2Z
and the like, which is selected according -to the kind of the protecting group on the amino group.
The decomposition by acid is one of the most suitable method and may be applied for the removal of the substituents e.g., benzyloxycarbonyl, substituted benzyloxycarbonyl, alkoxycarbonyl, substituted alkoxycarbonyl, aralkoxycarbonyl, adamantyloxycarbonyl, trityl, substi-tuted phenylthio, substituted aralkylidene, substituted alkylidene, substituted cycloalkylidene, and the like. The acid applied in-the above reaction is selected according to the kind of the amino protecting group, and the suitable acids are formic acid, trifluoroacetic acid, and the like, which are easily evaporated under reduced pressure. When the decomposition by acid i.s carried out in a solvent, a hydrophilic organic solvent, water or a mixture thereof is occasionally used as a solvent. The catalytic reduction may be applied for the removal of the amino protecting group e.g. benzyloxycarbonyl 9 substituted benzyloxy-carbonyl, 2-pyridylmethoxycarbonyl, and the like.
The suitable catalyst is palladium, and -the other catalysts conventionally applied for the catalytic reduction may be also used. The trifluoroacetyl group may be removed by treating the reaction product with water, and the halogen-substitu-ted alkoxycarbonyl group and 8-quinolyloxycarbonyl group may be removed by treating the reaction product with a heavy metal such as copper, zinc, and the like.

~al424Z2 The removing reaction of the amino protecting group may be carried out without the i~olation and the purification of the reaction product from the reaction medium.

,,, .

~314z4Z2 Some of the objective compounds of the formula :

1 ~ ~H-CONH ~

R2 ~N ~ CH2-R3' (I') wherein R'l is nitro group, alkanesul~onamido group, alXylaminosulfonamido group, alkylureido group or alkylthioureido group, R'3 i~ heterocyclic-thio group, in which the heterocyclic group may be ~ub~tituted with alkyl group, R2, X and M are as defined above, may be prepared by reacting 7-~ubstituted phenylgly-cinamido-3-alkanoyloxymethyl-3-cephem-4-carboxylic .
acids of the formula:- . . -1 ~3` 5 R2 ~ ~L CH2--0--R3"
OOM (I") wherain R"3i~ alkanoyl group, R'l~ R2, X an~ M are as defined above, with thiol compounds of the formula:
R'3 - H (IV) wherein R'3 is a~ defined above, or their alkali metal ~alts.

The term " alkanoyl group" represented by R"3 in the formula (I"~ means acetyl, propionyl, butyryl, and the like.
The alkali metal salt of the thiol compound (IV) :~
may be a ~odium ~alt, a potas~ium salt, or the like.

-- 10 _ ~042~ Z
The reaction of the 7-substi-tu.ted phenylglycinamido-

3-alkanoyloxymethyl-3-cephem-4-carboxylic acids (I") with the thiol compounds (IV) or the alkali metal salts thsreof may be carried out in a solvent, for example, water, acetone, chloroform, nitrobenzene, dime-thylformamide, methanol, e-thanol, dimethylsulfoxide, or any other organic solyen-ts inert to -the reaction, preferably in a strongly polar solvent. Among the solvents, hydrophilic solvents may be used in a mixture with water. The reaction is preferably carried out in around neutral medium. When the compound (I") or the thiol compound (IV) is used in a free form, the reaction is preferably conducted in the -presence of a base, Eor example, alkali metal hydroxide, alkali metal carbonate, alkali metal bicarbonate, trialkylamine, and the like. The reaction temperature is not restrictive, and the reaction is usually carried out at room temperature or under warming.
The reaction product can be isolated Erom -the reaction mixture by conventional methods.
When .the reaction product has the protected amino group ~or X, the amino protecting group may be removed from the reaction product by conventional method mentioned above, i:E necessary, and this process is also included in -the scope of the present invention.
All the reactants to be employed in the various processes of the present invention may be commercially available or be prepared by conventional methods well , , lO~'Z4~ZZ
known to the art or by a variety of analogou3 method~
applicable to production of ~uch reactant~, In accordance with the pr,e~ent inv~ntion, a precipatate which form~ during tha reaction is ~eparated from the reaction mlxture by methods commonly used for thi~ purpose, and the re~ulting reaction product may be subjected to routinely used purification procedure~, for instance, to recry~tallization from an appropriate solvent or a mixture of such solvents.
The compounds of the present in~ention may be converted by conventional r~lethods of forming ~alts from acids into their pharmaceutically acceptable, ~ubstantially non-toxic salts, for instance, by reaction with an alkali metal hydroxide, an alkali metal bicarbonate, an alkali metal carbonate, or an organic base, sodium salt being preferred. The preferred method of preparing the ~alts consists in dissolving ~he acid in a solvent wherein the salt is insoluble and then adding a ~olution of the salt-forming compound or the base thereto. Thereby thesalt precipitate~ from the reaction mixture.
The compounds of the present invention exhibit a high antibacterial activity and inhibit the growth of a number of microorganisms including Gram~
positive and Gram~negative bacteria. For therapeutic admini~tration the cephalosporin compounds according , . ~ ' ' ,': ' ' ' ' . ~ ' , . ~ .

~04~Z
to the present invention are used in the form of pharmaceutical preparation which contain 3aid compounds in admixture with a pharmaceutically acceptable organic or inorganic solid or liquid excipient quitable for oral or parenteral administra-tion~ ~he pharmaceutical preparations may be in ~olid form e.g., capsule~, tablet3, or dragees, or in liquid form e.g., solutions, suspensions, or emulsion~. If de~ired, there may be included in th~
above preparations auxiliary ~ub~tances, ~tabilizing agent~, wetting or emulsifying agents, buffers and other commonly used additives.
While the dosage of ~he compound~ will vary from and al~o depend upon the age and condition of the patient, an average single dose of about 100 mg., 250 mg., and 500 mg., of the compounds according to the pre~ent invention has proved to be effective in treating di~ease~ caused by bacterial infection. In general amount~ between 10 mg. and about 1000 mg. or even more may be administered.
The following examples are given for the purpose o~ illustrating the pre ent invention:-. . . ~

Examp7e 1. iO4~42Z
D-N-tert-Butoxycarbonyl-2-(3-methoxy-4-hydroxy-phenyl) glycine (2.97g.~ and triethylamine ~l.Olg.
were added to methylene chloride (50 ml.) and the mixture wa~ cooled at -lO to -15C. A methylene chloride ~olution (5 ml.~ containing pivaloyl chloride (1~17g~) was adcled to the mixture with ~tirring and the mixture was stirred at O to -10C for 2 hours. After cooling the resultant mixture to -20C, a methylene chlori~e solution (30 ml.) containing hydrochloride (3.34g.) of 2,2,2-trichloroethyl 7-amino-3-methyl-3-cephem~

4-carboxylate and 2,6-lutidine (1.94g.) wa~
added thereto all at once. The mixture was ~tirred at -10C for 1.5 hours and at room temperature for an hour. After removal of the ~olvent from the reaction mixture under reduced pres~ure, ethyl acetate (80 mlO) and 5% sulfuric acid (80 mlO) were added to the residue and the mixture was shaken~ The ethyl acetate layer wa~ 3eparated out, ~ashed in turn with 5% sulfuric acid (40 ml~), an aqueou~ 3aturated solution of sodium chloride, an aqueou~ sa~urated solution (40 ml.) of ~odium bicarbonate three times an*
- 25 finally, an aqueouY saturated solution of ~cdium chloride once, dried over magne~ium sulfate and then treated with activated charcoal. Removing the 301vent from the solution under reduced pre~3ure, tha re~idue wa~ pulverized with diisopropyl ether to give powder (4.86g.).
E - l - . ., ~
: . , .

iL~4Z42Z
The powder was purified by column chromatography on silica gel (120g.) [eluent: benzene: acetone

- 5:1] to give 2,2,2-trichloroethyl 7-[-D-N-tert-butoxycarbonyl-2-(3-methoxy-4-hydroxyphenyl) glycinamido]-3-mQthyl-3-cephem-4-carboxylate (3~58g.)~
I.R.

~ (cm 1) 3370, 1788, 1683 1740 ~.M.R.
CDCl3(ppm):1.41(9H,3), 2.17(3H,~), 3.30(2H,AB-q), 3.83(3H,s), 4.88(2H,AB-q), 4.94(1H,d,J=4Hæ), 5.15(IH,d,J=6Hz), 5.6~6.0(3H,m), lS 6.8~7~1(3H,m) Example 2.
2,2,2-Trichloroethyl 7-[D-N-tert-butoxycarbonyl-2-(3-methoxy-4~hydro~yphenyl)glycinamido] 3-methyl-3-cephem-4-c~boxylate(3.25g.), which wa~ obtained in a similar manner to that of Example 1, zinc powder (3g.) and glacial acetic acid (3 ml.) were added to dimethylformamide 530 ml.) and the mixture was stirred under ice-cooling for an hourO Filtering the reaction muxture, the filt-rate wa~ added to the mixture of ethyl acetate (60 ml.) and 3% aqueou~ hydrochloric acid (60 ml.) with ~tirring. After the resultant mixture was thoroughly sha~en, the ethyl acetate layer was separated out, washed with an aqueous saturated ~olution of ~odium chloride, dried over m~gnesium sulfate, and then concentrated under reduced * trademark. E - 2 - 15 ~

109~9L2~
pressure to glve 7-~D N-tert-butoxycarbonyl-2 ~3-methoxy-4-hydroxyphenyl)glyclnamido~
methyl-3-cephem-4-carboxylic acid (2.31g.).
I.R.

~ (cm 1) 3350, 1780, 1700, 1518 N.M R.
DMS0-d (ppm):1.38(9H,~), 1.97(3H,~), 3.40(2H,~Bq), 3.78(3H,~), 4.99(1H,d,J=4.5Hz), 5.21 ~ (lH,d,J=8Hz), 5.62(1H,dd,J-4Hz,7Hz), 6,6~7.2(4H,m), 8.93(1H,d,J-9Hz) Example 3.
7-rD-N-tert-Butoxycarbonyl-2-(3-methoxy-4-hydroxy-phenyl)glycinamido~-3-methyl-~-cephem-4-carboxy~ic acid (2.43g.) t w~ich was obtalned in a similar manner to those of Example~ 1 and 2~ was added to formic ~cid (20 ml.) and the mixture was stirred at room temperature for 3 hours. ~fter the formic acid wa~ removed ~rom the reaction mixture under reduced pressure, the re~idue wa~ pulverized with ether and 5% aqueous acetonitrile (20 ml.) wa~
added thereto. ~he re~ultant mixture was stirred !' ~or ~0 minute~ and then ~iltered to give pale yellow powder. The powder was washed with acetoni-trile and ether succes~ively and dried to give 7~-2-(3-methoxy-4-hydroxyphenyl)glycinamldo~^-3-me~hyl-3~cephem-4-carboxylic acld, m.p. 183 to 185C (dec.).
I.R.

~ (cm lj 2500 3500, 1775, 1697, 15709 ~Z 3L~2 N.M.R.
D20~DCl ~ppm~:2.10(3H,s), 3.36(2~,ABq~, 3.93(3H,~), 5.08(LH,d,J=4Hz), 5.32(1H,~), 5.70(lH,d,J=4Hz), 7,0~7.3~3H~m) W
pH6.4 phosphate buffer ~ max 237.3 m~, E=274 ~pH6.4 phospate buffer 266 m~, E=193 Example 4 DL-~-tert-Butoxycarbonyl-2-(3-methoxy-4-hydroxy~
phenyl) glycine (2.97g~) and triethylamine (l,llg,~
were added to tetrahydrofuran (25 ml.) and the mlxture wa~ cooled to O~C. Adding a tetrahydrofuran ~olution (S ml.) containing pivaloyl chloride (1.36g.) thereto, the resultant mixture wa~ stirred at 0C for 10 minutes. The solution of 7-amino~
3-methyl-3-cephem,4-carboKylic acid (2~14g.) ~-and triethylamine (l.Olg,) in 5~/0 aqueous acetone (30 ml.) wa~ added to the mixture all at once `
at 0C and stirred at room temperature for 1~5 hour~. Removing the organic solvent from the ~ reaction mixture under xeduced pressure, an aqueous saturated solution (30 ml.) of odium bicarbonate and ethyl acetate (50 ml,) was added to the residue. The aqueous layer was adjusted to pH2 with l~o aqueous hydrochloric acid, and then the ethyl acetate layer was separated.

.

-~ 4~22 The aqueou3 layer was extracted with ethyl ace-tate (50 ml.) twice, and this extract wa~ co~
bined with the previousl~ obtained ethyl aceta-te solution. After the solution wa~ dried over magne~ium sulfate, the ethyl acetate was removed under reduced pressure. The re~idue was pul-verized with dii~opropyl ether to give 7-[DL-N-tert-bu~oxycarbonyl-2-(3-methoxy-4-hydroxy-phenyl)glycinamido]-3-methyl-3-cephem-4-carboxylic acid (3g.), m.p. 127 to 130C (dec.) I.R.

~J (cm 1):3350,2650,1770, 1718,1695,1680, 1518,1275,1250,1160,1055,1032 860,782,720 Exa~ple 5.
7_ ~DL-N-tert-Butoxycarbonyl-2- ( 3-methoxy-~
hydroxyphenyl) glycinamido]-3-methyl-3-cephe~
4-carboxylic acid (2.0g.~, which was o~tained in ~;
a similar manner to hat of Exarr~?le 4, wa~ dis-solved in formic acid (20 ml.) and the ~olution wa~ qtirred at 40C for 4 hours. Removing formic acid from the ~olution under reduced pressure, the residue was pulverized with acetonitrile.
The powder was added to lOS/o aqueou~ acetonitrile, triturated with a glass-bar to pulverize, and then filtered. Thus obtained powder was washed with acetonitrile and ether succes~ively, and dried to give 7-[DL-2(3-methoxy-4-hydroxyphenyl)-glycinamido]-3-methyl-3-cephe~4-carboxylic acid (l.lOg.), m.p.193 to 197C (dec.).

.
.

, .

~C~4Z~Z~
I .R .

1~ i (cm~l):1760 1690 W

pH6~4phosphate buffer ~m~) 237, E=275 max pH6.4phosphate buffer max ~m~) 265,5, E=189 Example 6 D-N-tert-Butoxycarbonyl-2-~3-ni~ro-4-hydroxyphenyl) glycine (5g.) was dissol~ed in absolute methylene chloride (100 ml.). 2,6-Lutidine (1.9g.) was added to the solution and the mixture wa~
cooled at -10 to -15C~ Pivaloyl chloride (1.9g,) was dropwise added thereto and the -~
mixture was ~tirred at -15C for 3 hours. To .
- this mixture, was added all at once, at -5 to -10CJ a solution prepared by adding ~,0-bis ~trimethylsilyl)acetamide(10 ml.) to a suspension --of 7-amino-3-methyl-3-cephem~4-carboxylic acid (3.45g.) and methylene chloride (50 ml.) and stirring the mixture at room temperature for 3 hours. Thus obtained mixture was stirred at -10 to -15C for 3 hours and the solvent removed under reduced pre~aure. Ethyl acetate and 5%
sulfuric acid were added to the residue, and the ethyl acetate layer was separated out. To ~he ethyl acetate extract was added an aqueous qolution o~ sodium bicarbonate and the aqueous layer wa~ ~eparated out. The aqueous solution was acidified with ~ulfuric acid and bac~-extracted with ethyl acetate. The ethyl acetate , ~09~22 extract was wa3hed with water and an aqueous 3aturated solution of 30dium chloride successively and dried o~er magnesium ~3ulfate.
Removal o~ the ~olvent from the ~olution und~r reduced pressure gave powder (6~0g.) of 7 CD-N-tert-butc~xycarbonyl~2-(3~nitrc~4-hydroxyphenyl~glycinamido]-3-methyl-3~cephem-4-carboxylic acid, m.p. 150 to 155C. (dec.).
I.R.
Nujol ~J (cm 1):1755, 1700, 1690, 1670 ~oM~R ~
DMSO~d (ppm):1.40(9H,s),2.03(3H,s), 3.38(2H,broad s),4.99(1H, d,J--4.5Hzl,5.32(1H,d,J=8.5Hz~, 5.60(H,dd,J=4.5,8.0Hz), 7.10(1H,d,J=8Hz),7.2^-7.55 (lH,m),7.65(1H,dd,J=2.0, 8.0Hz),8.05(~H,d,J=2Hz),9,13 (lH,d,J=8Hz) Example 7 7--~D-~-tert-Butoocycarbonyl)-2-(3-nitro-4-hydroxyphenyl~glycinamido]-3-methyl-3-cephem-4-carboxylic acid (5g.), which was obtained in a similar manner to that of Example 6, was dis-solved in formic acid (40 ml.) and the solution was stirred at room telT~erature for 4.5 hours.
Removing the solvent from the solution under reduced pres~ure, the residue was pulverized with ether and filtered. Thus obtained powder was wa3hed with ether, ethyl acetate and ~IL04;~42Z
acetonitrile successively and then suspended in acetonitrile (40 ml.). Water (4 ml.) was dropwise added to the suspension under stirring and the mix-ture was stirred at room temperature o~ernight. The appearing crystals were ~iltered, washed with acetonitrile ~nd dried to give 7-[D-2-(3-nitro-4-hydroxyphenyl)glycinamido)-3-methyl-3-cephem-4-carboxylic acid (2.64g.), m.p. 170 to 172C. (dec.).
I.R.

~ (cm~l)o3200, 1763, 1702, 1628, 1546 - N.M.R
D O+DCl 2 (ppm):2.15(3H,s),3~4(2H,d,J=4Hz), 5.13(1H,d,J=4.5Hz),5.54(1H, s),5.70(1H,d,J=4.5Hz), 7.38(1H,d,J=8.5Hz~,7.91 (lH,dd,J=2.5,8.5Hz),8.45 ~lH,d,J=2.5Hz) W ' :
pH6.4phosphate buffer (m~) 242, E=434 max 25 Example 8 To a solution of D-N-tert-butoxycarbonyl-2-nitro-~-hydroxyphenyl)glycine (lg.) in methylene chloride (20 ml.), was added 296-lutidine (0.38g.) and the mixture was cooled at -15 to -20C. A methylene chloride solution (1 ml.) of pivaloyl chloride (385 mg.) was dropwise added thereto and the mixture was stirred at -20C for 2 hours. To this ,.. . . .

: ., , : .. .. . :-. ~. . .

mixture wa~ added at -20C all at once a solu-tion prepared by adding 2,6-lutidine (0.64g.) to a suspension of p-toluene sulfona-te (1.9lg.) of 2,2,2-trichloroe-thyl 7-amino-3-methyl-3-cephem-4-carboxylate in methylene chloride (20 ml.) under ice-cooling and stirring thus obtained mixture for 15 minutes. Stirring the resultant mixture at -20C for 3.5 hours, the solvent was removed there~rom under reduced pressure. Ethyl acetate and water were added to the residue and insoluble substances were filtered off. The filtrate was washed with ethyl acetate to give 2,2,2-trichloroethyl-7-(D-N-tert-butoxycarbonyl-2-(3-nitro-4-hydroxvphenyl)glycinamido~-3-methyl-3-cephem-4-carboxylate (0.27g.). After the filtration of the product, the ethyl acetate layer and the washings of eth~l acetate were combined, washed with 5% hydrochloric acid, an aqueous saturated solution of sodium bicarbonate, and water in turn, and dried over magnesium sulfate, and then the solvent was removed there~rom under reduced pressure. The resultant gel-like residue was washed with ether to give the same object product (1.09g.). Total yield 1.36g~
I.R. -~

Nujol (cm~ ):3320, 32703 1775, 1735, .

16859 1650, 163 N.M.R.

- 22 - ~ -. .. ~ - . - : -. . -24;~Z
~DMSO-d6 (ppm):1.40(9H,s),2.10(3H,s),3.49(2H, broad s),5.05(2H,ABq),5.07(1H, d,J=4.5Hz),5.35(1H,d,J=8Hz), 5.69(1H,dd,J=4.5Hz,8.0Hz), 7.10(1H,d,8.5Hz),7.2-7.5(1H, ~-m),7.63(1H,dd,J=2.0,8.5Hz), 8.04(1H,d,J=2Hz),9.2(1H,d,J=
8Hz) ;

Example 9.
To a suspension of hydrochloride (3820 mg ) of 2,2 7 2-trichloroethyl 7-amino-3-methyl-3-cephem-4-carboxylate ir. absolute methylene chloride (50 ml.~, were added dropwise -triethylamine (810 mg.) and N,N-dimethylaniline (245 mg.) under ice-cooling with stirring. The mixture was stirred at room -temperature for 30 minutes. N-tert-Butoxycarbonyl-2-(3-mesylaminophenyl)-D-glycine (3440 mg.), triethylamine (1010 mg.) and N,N-20 ~ dlmethylbenzylamine (4 drops) were added to absolute methylene chloride (50 ml.) and the mixture was stirred under dry ice-acetone cooling. A solution ~
of ethyl chloroformate (1085 mg.) in absolute~ .
methylene chloride (25 ml.) was dropwise added -to the mix$ure at -25 to -30C in lO minutes. Thi~
mixture was stirred at the sam8 temperature for;~
15 minutes. To the mixture, was added all at once the previously obtained mixture precooled at -15C. The resultant mixture was stirred a-t ~Q -25 to -30C ~or`4 hours, and washed with water, - 23 ~

.. , ... ~

3~ hydrochloric acid and water respectively.
The organic layer was separated out, washed with 3~ aqueous solu-tion of sodium bicarbonate and water in turn, and dried. The methylene chloride was removed under reduced pressure, and the residue was dissolved in e-thyl acetate (10 ml.). Ether was added thereto and the appearing crystals was ~iltered to give 2,2,2-trichloroethyl 7-[D-N-ter-t-butoxycarbonyl-2-(3-mesyoaminophenyl)glycinamido] -3-methyl-3-cephem-4-carboxylate (4195 mg.), mp 204 to 205C (dec.).

The above obtained 2,2,2-trichloroethyl 7-[D-N-tert-butoxycarbonyl-2-(3-mesylaminophenyl)-glycinamido]-3-methyl-3~cephem-4-carboxylate (3985 mg.) was dissolved in dimethyl~ormamide (17 ml.). To the solution were added acetic acid , (5.0 ml.) and zinc powder (3985 mg.) under ice cooling with stirring and the mixture was stirred at the same temperature for 2 hours. After an insoluble product was filtered o~f, the filtrate was washed with dimethyl~ormamide (3 ml.). The ;
washings and the previously obtained filerate were combined together and added to 5% hydrochloric acid (100 ml.) under ice cooling. Water (50 ml.) was added thereto and the resultant mixture was extracted three times with ethyl acetate (50 ml.).
The extract was washed with water and back-extracted three times with 5% aqueous solution (50 ml.) o~

E ~
'~:

~ 24 -1~24;~Z
sodium bicarbonate, and the aqueous layer was acidified with hydrochlori c acid. The aqueous ! solution was back-extracted again with ethyl acetate, and the extract was washed with water ~ - -and dried. The solvent was removed under reduced pressure, and the residue was dissolved in ethyl ~;
acetate (10 ml.). The solvent was allowed to stand at room temperature ~or an hour, and ether was added to the solution to give crystals 7-[D- ;
N-tert-butoxycarbonyl-2-(3-mesylaminophenyl)-glycinamido]-3-methyl-3 cephem-4-carboxylic acid (2687 mg.), m.p. 187 -to 189C (dec.).

Example 10.
~;~ To a suspension of hydrochloride (26.8g.) of 2, 2,2-trlchloroethyl 7-amino-3-methyl-3-cephem-4~
~; carboxylate in dry methylene chloride (500 ml.) 7were added successively a solution of triethylamine !(7g~) ln dry methylene chloride (25 ml.) and then a solution of 2,6-lutidine (2.14g.) in methylene 20 ~ ~; chloride ~(25 ml.~ u~der ice cooling. And ~urthermore, ;~
N-tert-butoxycarbonyl-2-(3-mesylaminophenyl)-D-glycine (26.0g.) and then dicyclohexylcarbodiimide (15g.) were gradually added thereto and the mixture was stirred under ice cooling for 1.5 hours.
A~ter removal of an insoluble product by ~iltration, -~
the filtrate was washed ~our times with ice-; cooled 5% hydrochloric acid (100 ml.), on~e with water, three times wi-th lO~o aqueous solution of sodium bicarbonate and once with an aqueous 3~09~24Z~:
- saturated solution of sodium chloride, respectively, and then dried over magnesium sulfate. The solution was treated with activated charcoal and the solvent was removed under reduced pressure. The residue Was dissolved in ethyl acetate (100 ml.) - and allowed to stand at room -tempera-ture overnight.
The7appearing crystals were filtered to give colorless plates of 2,2,2-trichloroethyl 7-[D-N-tert-butoxycarbonyl-2-(3-mesylaminophenyl)-glycinamido~-3-methyl-3-cephem-4-carboxylate (31.3g.), m.p. 189 to 191C (dec.).

The above obtained 2,2,2-trichloroethyl 7-[D-tert-butoxycarbonyl-2-(3-mesylaminophenyl)-glycinamido)-3-methyl-3-cephem-4-carboxylate ~12.01g~) was dissolved in dimethylformamide (40 ml.). Glacial acetic acid (15 ml.) and zinc powder (12g.) were successively added to the solution under ice cooling and the mixture was kept stirred at the same temperature for an hour.
After the reaction was over, an insoluble product was fil-tered and washed with ethyl acetate. The filtrate and the washings were poured to 3%
hydrochloric acid (300 ml.) under ice cooling.
.
The mixture was extracted three times with ethyl acetate (150 ml.). The ethyl acetate extract was washed with water and then back-extracted three times with 5% aqueous solution (150 ml.) of sodium bicarbonate. The aqueous solution was washed with ethyl acetate and adjusted E ~ 13 :.,, . . ,,. ~ . , .
: ~ ., .,.. ' - ,. ': .
: . . .

~)4~:~L2;Z ~
~o pH2 with 10% hydrochloric acid. The appearing product was extracted -three times with ethyl acetate (150 ml.). The extract was washed with water and dried over magnesium sulfate, and the solvent was removed under reduced pressure. The appearing crystals were washed with ethyl acetate and ether, respectively, to give 7-~D-N-tert-butoxycarbonyl-2-(3-mesylaminophenyl)-glycinamidol-3-methyl-3-cephem-4-carboxylic acid (8.38g.), m.pO 188 to 189C (dec.).

I.R.

(cm 1) 3330, 3290, 3240, 1769, 1720, 1690, 1668, 1524, 1308, 1220, lS 1146, 9129 890, 772 N.M.R. ~; -~DM~O-d6 (ppm):1.40(9H,s),2.00(3H,s), ;
2.99(3H,s),3.37(2H,broad s) 9 ~ 4.96(1H,d,J=4.4Hz),5.30(1H, broad d,J=8Hz),5.63(lH,d,d, J=4Hz,7Hz),6.90~7.50(4H,m) The above ohtained 7-[D-N;-tert-butoxycarbonyl-2-(3-mesylamino~glycinamido]-3-methyl-3-cephem-4-carboxylic acid (8.28g.) was added to formic acid (140 ml.) under ice cooling and then the mixture was stirred at room temperature for 1.5 hours. Removing the formic acid at 35C

.

~04Z~ZZ
under reduced pressure, the residue was dissolved in 5~ hydrochloric acid (30 ml.). The solution was washed wlth ethyl acetate (20 ml. ), treated with ac-tivated charcoal, and then ad~usted to pH3 with 10% aqueous solution o~ sodium hydroxide.
The appearing crystals were filtered, washed with water and dried to give 7-[D-(3-mesylaminophenyl)glycinamido~-3-methyl-~-cephem-4-carboxylic acid (6.18g.), m.p. 199 to 199.5C (dec.).

~C(~2 = +131 (o.lN-Hcl~ C=l) U.V.
pH6.4 phosphate buffer 15 max 263.5 m~, E=172 ;;
pH6.4 phosphate buffer -,~
in~ 228 m~, E=Z99 ~-N.M.R.
20 D20~DCl (ppm~:2.08(3H~s),3.19(3H,s), 3.2193.44(2H,A~q,J=18Hz), 5 04~1H,d,J=5Hz),5.37(1H,s~
5.67(1H,d,J=5Hz),7.4~7.67(4H, m) I.R.

(cm ):3610, 3510, 3330, 17509 1700, 1600, 1526, 1380, 1366, 13289 . .
., , .. . - . - . . ;

: . - ~ . ~ , . . .

Example 11. ~ ~ Z~ ~ ~
A solution of ethyl chloroformate (1.~2g.) in dry methylene chloride (20 ml.) was cooled to -10C. To this solution was added dropwise in 10 minutes a solution of N-tert-butoxycarbonyl-2-(3-ethanesulfonamidophenyl)-D-glycine (4.4g.) and triethylamine (1.22g.) in methylene chloride (20 ml.) and N,N-dimethylbenzylamine (2 drops), and then the mixture was stirred at room temperature for an hour. On the other hand, 27 2,2-trichloroethyl 7-amino-3-methyl-3-cephem-4-carboxylate (3.82g.), triethylamine (0.9g.) and N,O-bis(trimethylsilyl)acetamide ~0.12g.) were dissolved in absolute methylene chloride (40 ml.).
The solution was dropwise added to the above obtained ~ixture at -15C in 10 minutes. The ~ ,:
resultant solution was kept stirred at the same temperature for 2 hours and washed twice with 2%
hydrochloric acid, ~n aqueous solution of sodium bicarbonate and an a~ueous saturated solution of sodium chloride respectively, and dried over ~ ~-magnesium sulfate~ Removing the solvent ~rom the solution under reduced pressure, the residue was pulverised with a little of ethanol to give colorless crystals (4.2g.) of 2,2,2-trichloroethyl 7- LD-N-tert-butoxycarbonyl-2-(3-ethanesulfonamidophenyl)-glycinamidol-3-methyl-3~cephem-4-carboxylate, m.p. 117C.

.. . . . . . ..

.,~" ~: , .. .

I.R.

~ (cm 1~ 3400, 3230, 1762, 1735, 1690 N.M.R.
~CDC13 (ppm):1.28~3H,t,J=7Hz),1.38(9H,s), 2.16(3H,s),2.75w3.50(4H,m), 4.82,4095(2H,ABq.J=12Hz), 4.95(1H,d,J=5Hz),5.24(1H,d,J=
6Hz),5.81(1H,d,d,J=5Hz, J=8Hz),7.25(4H,s).
~ .
The above obtained 2,2,2-trichloroethyl 7-[D-N-tert-butoxycarbonyl-amino-2-(3-ethanesulfonamidophenyl-glycinamido~-3-methyl-~-cephem-4-carboxylate ~4.2g.) was dissolved in a mixture of absolute dimethylformamide (15 ml.) and acetic acid ~
(45 ml.). Zinc powder (3.6g.) was added to the ~; -solution under ice cooling and the mixture was ~ ~-stirred for 2 hours. After the zinc powder was filtered off, the ~iltrate was poured into - the mixture of 2% hydrochIoric acid (40 ml.) and ethyl acetate (40 ml.) and the ethyl acetate layer was separated out. The aqueous layer was extracted further with ethyl acetate (20 ml.).
The ethyl acetate layer and extract were combined together; washed with 2% hydrochloric acid (20 ml.) and an aqueous saturated solution of sodium chloride (20 ml.) in turn and dried over ; ~ - 17 ... ... . . .

~(~42~L~%
magnesium sulfa-te. The solvent was removed from the solution under reduced pressure and then the residue was washed wi.th diisopropyl ether to give oily product (3.2g.) of 7-~D-N-tert-butoxycarbonyl-2-(3-ethanesulfonamidophenyl)-glycinamido~-3-methyl-3-cephem-4-carboxylic acid.

I.R~
film ~ (cm~l):3300, 17~5, 1700 N~M~Ro CD OD
(ppm):1.22(3H,t,J=7.5Hz),2.09(3H,s), 2.75~3.75(4H,m),4.96(1H,d~J=
4.5Hz),5.26(1H9s),5.67(1H, d,J=4.5Hz),7.25(4H,s).

A solution of the above obtained 7-[D-N-tert~
butoxycarbonyl-2-(3-ethanesulfonamidophenyl)-glycinamido~-3-methyl-3-cephem-4-carboxylic ~ ;
acid (3.1g.j in formic acid (15 ml.) was stirred at room temperature for 2 hours. The -~
formic acid was removed from the solution at room temperature under reduced pressure. The residue was pulverized with ether and filtered.
The resultant powder was added to acetonitrile (10% aqueous solution) (20 ml.), stirred under ice cooling for an hour and then the appearing crystals were filtered to give white crystals (1.7g.) of 7-[D~2-(3-ethanesulfonamidophenyl)-.. . .

,. . . : ~- . . .

10~ 2;2~
glycinamido)-3 methyl-3-cephem-4-carboxylic acid, m.p. 179 to 182C (dec.).

I.R.
Nujol 1) (¢m~ ):1770, 1695, 1600 N.M.R.
D O~NaHC0 2 3(ppm):1.29(3H,t,J=7.5Hz),l.90 (3H,s),2.75~3.67(4H,m), 4.9~(1H,dJJ=4.5Hz),5.25 - (lH,s),5.59(1H?d,J=4.5Hz), 7.34(4H,s) Example 12.
........... ., .:
2,2,2-Trichloroethyl 7- D~N-tert-butoxycarbonyl~
2-(3-mesylaminophenyl)glycinamido)-3-methyl-3- -~
cephem-4-carboxylate (l.Og.) was added to ice-cooled formic acid (20 ml.) and the mixture was stirred at room temperature for 2 hours. Removing the formic acid from the mLxture under reduced pressure, water was added to the oily residue.
. The mixture was adjusted to pH8-9 with an aqueous saturated solution o~ sodium bicarbonate under ice cooling. The appearing crystals were :Eiltered, washed with water and then dried over phosphorus pentoxide to ~;ive 2,2,2-trichloroethyl 7-~D-2-(3 mesylaminophenyl)glycinamido3-3-methyl-3-cephem-4-carboxylate (0.78g.)j m.p. 107 to 110C (dec.).

I.R. 104Z4ZZ
Nujol ~ (cm~l):~350, 1785, 1740, 1685 Example 13.
To a suspension of hydrochloride t5.9g) of 2,2,2-trichioroethyl 7-amino-3-methyl-3-cephem-4-carboxylate in methylene chloride (100 ml.
were added a solution of triethylamine (1.55g.) in methylene chloride (10 ml.) and a solution o~
2,6-lutidine (0.16g.) in methylene chloride (10 ml.). N-tert-Butoxycarbonyl-2-(4-mesylaminophenyl)-D-glycine (5.8g.) and dicyclohexylcarbodiimide (3.3g.) were added to the resultant solution under ice cooling. The mixture was kept stirred at the same temperature for 3 hours, and filtered. The ~iltrate was concentrated under reduced pressure, and ethyl acetate (200 ml.) was added to the residue. The mixture was washed with 5~ hydrochloric acid, water, ;
20 ; ~an aqueous saturated solution,of sodium ` bicarbonate and water, respectively, dried and then concentrated to give 2,2,2-trichloroethyl 7-[D-N-tert-butoxycarbonyl-2-(4 mesylaminophenyl)-glycinamido3 -3-methyl-3-cephem-4-carboxylate ~10.83g.), m.p. 129 to 136C (dec.).
~ ':
I~R.

LJ j (cm~l):3290 1770 1680 ~L~42~ZZ
N.M.R.

(ppm):1.43(s,9H),2~20(s,3H), 3.~2(AB-q,2H),4.8~5.0(m,3H), 5.2~6.0(m,3H),7.0~7.7(m,4H)~
7.9(broad s,lH) Acetic acid (12.5 ml.~ and zinc powder (lOg.) were added to a solution of 2,2,2-trichloroethyl 7-[D-N-tert-butoxycarbonyl-2-(4-mesylaminophenyl)- ~ ;
glycinamido]-3-methyl-3-cephem-4-carboxylate (lQg.) in dimethylformamide (33 ml.) under ice cooling. The resultant mixture was stirred at the same temperature for an hour and then filtered. The filtrate was added to a mixture of 5% hydrochlori¢ acid (lOQ ml.), iee water t50 ml.) and ethyl acetate (100 ml.), and then extracted three times with ethyl acetate (100 mlO). The extract was back-extracted twice with 5% aqueous solution of sodium bicarbonate (100 ml.). The aqueous layer was washed with ethyl acetate 7 adjusted to pH2 with 10%
hydrochloric acid and then extracted with ethyl acetate. The ethyl acetate extract wa9 washed with water and dried and then the solvent was removed under reduced pressure to give 7-[D-N-tert-butoxycarbonyl-2-(4-mesylaminophenyl)-glycinamido~-3-methyl-3-cephem-4 carboxylic -~
aci~ t-7.8g;), m.~. 180 to 200C (dec.).

I.R.

. - . , . . :. , . .. ~ ",, .

~1~42~a~2 Nujol (cm 1):3300, 1770, 1710(shoulder), 1695, 1680 N.M.R.
~DMS0-d6 (ppm):1.40(s,9H),2.02(s,3H),2095(s,3H), 3.2~3.7(m,2H),4.97(d,1H),5.2^~.8 (m,2H),7.30(AB-q,4H) 7-[D-N-tert-Butoxycarbonyl-2-(4-mesylaminophenyl)-glycinamido~-3-methyl-3-cephem-4-carboxylic acid (4. 6g.) was added to formic acid (70 ml.) under ice cooling. The resultant mixture was kept stirred at the same temperature ~or 2 hours and then concentrated under reduced pressure and water was added to the residue. The resultant aqueous mixture was washed with ethyl acetate, adjusted to pH6 with an aqueous solution of sodium bicarbonate, concentrated to one hal~ of its initial volume, absorbed on a resin absorbance (trade mark:Amberlite XAD-2) (460g.), ~ ;~
20 ~ which was prewashed with methanol and water, and then eluted with water and miethanol. The eluate was concentrated and the appearing crystals were :~ filtered and washed with methanol to give 7-[D-2-(4-mesylaminophenyl)glycinamido~-3-methyl-3-cephem-- 4-carboxylic acid (2.1g.), m.p. 205 to 207C (dec.).

I.R.
Nu301 ~ (cm~l):3175, 1760~ 1670 4~2 N . M . R .

(ppm):1.80(3H,s),3.05(3H,s),3.15(2H, AB-~),4.85(1H,d),5.12(1H,s), 5.54(1H,d),7.35(L~AB-q) Example 14.
N-tert-Butoxycarbonyl-2-(3-mesylaminophenyl)-D-glycine (2.066g.), triethylamine (0.606g.) and N,N-dimethylbenzylamine (15 ml.) were added to tetrahydrofuran (20 ml.) and the mixture was cooled to -10 to -12C. A solution of isobutyl chloroformate (0.820g.) in tetrahydrofuran (10 ml.) was dropwise added thereto at the same temperature in 2 minutes, and then the resultant mixture 15 - was kept stirred at the same temperature ~or 30 minutes. On the other hand, 7-amino-3-methyl-3-cephem-4-carboxylic acid (1.070g.~ and triethylamine (0.581g.) were added to 50%
aqueous tetrahydrofuran (30 ml.) under ice cooling. m e solution was added to -the above obtained mixture, which was Gooled to -6C, all at onçe. Thus obtained mixture was stirred ~ under ice cooling for an hour and additionally at room temperature for 2 hours, after which the tetrahydrofuran was removed under reduced pressure. An aqueous saturated solution (15 ml.) of sodium bicarbonate was added to the residue and the resultant aqueous washed twice with ethyl acetate (10 ml.). The washings were extracted with an aqueous saturated solution of sodium bicarbonate (10 ml.). ThP aqueous extract was - , - - . . :, , ` , . .
: -,: , , .: .
. ~ .

lO~LZ4Z2 combined wi-th the above obtained aqueous solution and then ethyl acetate (30 ml.) was ~urther added on the combined aqueous solution. 10% hydrochloric acid was added thereto to adjust i-t to pH2.
The aqueous layer and et;hyl acetate layer were s~aken together thoroughly and an insoluble produc-t was filtered off. The ethyl acetate layer was separated out and the aqueous layer was extracted twice with ethyl acetate (20 ml.).
Thus obtained extract and the ethyl acetate layer obtained previously were combined together and washed with water (10 ml.). The aqueous washings were extracted twice with ethyl acetate (5 ml.) and thus obtained extract was combined with the ~;
ethyl acetate solution obtained above. The mixed solution was washed with an aqueous saturated solution (10 ml.) of sodium chloride, dried over magnesium sul~ate and treated with activated charcoal, and the removal of the solvent gave ~ pasty residue (3.47g.). The residue (3.42g.) was added to absolute ether (30 ml.), and the mixture was stirred at room temperature overnight.
The appearing crystals were filtered, washed with ether and dried to give 7-[D-N-tert-butoxycarbonyl-2-~3-mesylaminophenyl)glycinamido~-3 methyl-3-- cephem--4~carboxy1ic acid (2;326g.), m.p. 174C
(dec.). ;

~D = 57 (C=l, methanol) ~' . . ' . ' ! ' F~ample 15. lV4Z42Z
N-tert-Butoxycarbonyl-2-(3-mesylaminophenyl)-D-glycine (3.44g.) and triethylamine (l.Olg.) were dissolved in absolute methylene chloride (Z5 ml.).
The solution was dropwise added to the solution o~ isobutyl chloroformate (1.36g.) in absolute meth~lene chloride ~35 ml.) at -10 to -15C in 5 minutes and stirred at the same temperature ~or 15 minutes. On the other hand, N,O-bis(trime-thylsilyl)acetamide (3.5g.) was dissolved in the suspension of 7-amino-3-(5-methyl-1,3,4-thiadiazol-2-yl)thiomethyl-3-cephem-4-carboxylic acid ~3.44g~) in absolute methylene chloride (30 ml.). Thus obtained mixture was dropwise added to the above prepared solution o~ mixed anhydride at -15C, and then stirred at the same temperature for 1.5 hours and, additionally at 10C ~or 3 hours. The resultant mlxture was washed with 5% hydrochlori~ acid and water respectively, dried and then the solvent was :`
removed. The oily residue was purified by column chromatogra~hy on silica gel (eluent:
~:~ chloroform) to give oily product of 7-[D-N-tert- ;:
butoxycarbonyl-2-(3-mesylaminophenyl)glycinamido]-3-(5-methyl-1,3,4-thiadiazol-2-yl)thiomethyl-3-cephem-4 carboxylic acid (3.8g.).

I.R.
~: film ! : `
(cm~l):3300, 1780, 1725, 1685, 1670 ~:

11~4;~4~2 Thus obtained 7-[D-N-tert-butoxycarbonyl-2-(3-methylaminophenyl)glycinamido]-3-(5-methyl-1,3, 4-thiadiazol-2-yl)thiomethyl-3-cephem-4-carboxylic acid t2.23g.) was dissolved in formic acid (35 ml.) and stirred at 18-20C for 4 hours.
The resultant mixture was concentrated under reduced pressure, pulverized with ethyl acetate and then filtered to gi~e 7-[D-2-(3-mesylaminophenyl)-glycinamido3-3-(5-methyl-1,3 7 4-thiadiazol-2-yl)-thiomethyl-3-cephem-4-carboxylic acid (1.95g.).
The product was added to acetone (30 ml.), stirred at 15-20C for an hour, allowed to stand and then the supernatant solution was removed by decantation. Acetone (30 ml.) was added to the residue, stirred at 15-20-C for 3 hours and then the precipitate was ~iltered off.
The precipitate was washed with acetone and ether, respectivel~, to give pale yellow powder of the pure object product.

I.R.

Nujol lJ (cm~l):1765, 1690, 1605 N.M.R.

r D2o-Dcl (ppm):2.97(3H7s~,3.01(3H,s),3.38, 3.78(2H,AB-q,J=18Hz),4.26, 4.47~2H,AB-q,J=14Hz),5.12(1H, d,J=4Hz),5.35(1H,s),5.71(1H, d,J=4Hz) _ ~9 _ .. . . .
. . .
.,. , ~
;........... l , . - . , :

Example 16. 1~42422 (1) Dicyclohexylcarbodiimide (2.0g.) was added under ice cooling with stirring to a solu-tion of N-tert-butoxycarbonyl-2-~3-mesylamino-4-hydroxyphenyl)-D-glycine (5.2g.), hydrochloride (5.52g.) of 2,2,2-trichloroethyl 7-amino-~-methyl-3-cephem-4~carboxy1ate and 2,6-lutidine (1.74g.) in absolute methylene chloride (180 ml.). The resultan-t mixture was kept stirring at the same temperature for an hour, and additionally, at room temperature for 3 hours. An insoluble product was filtered off and the filtra-te was condensed under reduced pressure. Ethyl acetate was added to the residue and then the solution was adjusted to about pH2 with phosphoric acid. The ethyl acetate layer was separated out, washed with water and dried over magnesium sulfate. Removing the ethyl acetate from the solution under reduced pressure, the residue was pulverised with isopropyl ether to give 2,2,2-trichloroethyl 7-tN-tert-butoxycarbonyl-2- ;
(3-mesylamino-4-hydroxyphenyl)-D-glycinamido¦-3-methyl-3-cephem-4-carboxylate (6.75g.). The product was crystallized from ethyl acetate to give the purified product~ m.p. 185 to 188.5C (dec.).

I.R.
Nujol (c~ 1766 N~M.R.

_ 40 -~ ~ .
~ . .. :. . ' ' . . . . . :

~O~Z42Z
( 3)2 (ppm):1.40(9H,s),2.16(3H,s),3.01 (3H,s)~3.34,3.59(2H,AB-q, J=:L8Hz),4.87,5.07(2H,AB~q, J=12.5Hz),5.07(1H,d,J=4.5Hz), 5.35(1H,d,J=8Hz),5.80(1H,d,d, J_4.5Hz),8Hz),6.3-6.5(1H,m)~

6.90(1H,d.J=8Hz),7.20(1H,d,d, J=ZHz,8Hz),7.52tlH,d,J=2Hz), 8.0-8.3(2Hgm) (2) Acetic acid (3.5 ml.) and zinc powder (2.6g.) were ~;
added under ice cooling to a solution o~ Z,2,2-trichloroethyl 7-(N-ter-t-butoxycarbonyl-2-(3-- -- mesylamino-4-hydroxyphenyl)-D-glycinamido~-3-~5 methyl-3-cephem-4-carboxylate (3.0g.) in dimethylformamide (9 ml.). m e mixture,was kept stirring at the same temperature for 40 minutes, After the reaction was over, the zinc powder was ~iltered off and washed with a little of r dimethylformamide, and then the filtrate and - the~washings were combined together. Ethyl acetate and dilute phosphoric acid were added to , .
the mixture and the ethyl acetate layer was separated out. The extract was washed with water and back-extracted with an aqueous solution of sodium bicarbonate. The aqueous solution was - acidified with hydrochloric acid and then extracted with ethyl acetate. The ethyl acetate extract was washed with water and dried over magnesium sulfate and the solvent was removed under reduced pressure. The residue was washed with ether to .. ~ , , .
~: ~ '' ' ' ' ' " ' ~ -- . . , ~242;~
give powdery product (1. 5g.) of 7-(N-tert-butoxycarbonyl-2-(3-me~ylamino-4-hydroxyphenyl)-D-glycinamido]-3-methyl-3-cephem-4-carboxylic acid.

I.R.
Nujol V (cm 1):1765 N.M.R.
D20+NaHCo3 ~ (ppm):1.40(9H,s),1.90(3H,s), ~ -3.05(3H,s),3.00,~.39(2H, AB-q,J=18Hz),4.90 (lH~ d, J=4.5Hz),5.08(1H,s), 5~52(1H~d~Ja4.5Hz)~6.8- ~;

- 7.4(3H,m) (3) A solution of 7-[N-tert-butoxycarbonyl-2-(3-mesylamino~4-hydroxyphenyl)-D-glycinamido~ 3-me~hyl-3-cephem-4-carboxylic acid~ (1.45g.) in ~ormic acid (6 ml.) was stirred at 40C for 1.5 hours. After reaction was over, the acetic acid 20 ~ was removed under reduced pressure and then acetonitrile (30 ml.) and water (0.5 ml.) were added to the residue with stirring and the mixture `~
was stirred for 30 minutes. The appearing powder was filtered and washed with acetoni-trile and ether, respectively to give 7-[2-(3-mesylamino 4-~; hydroxyphenyl)-D-glycinamido~ methyl 3-cephem-4-carboxylic acid ~1.13g.~, m.p. 186 to lg2C
(dec.).
,.

~;

',' ';, ' ' ' . , ', , : ' ! , I I , ' ~0~L24~2 I.R.

~ (cm 1):1760 N.M.R.
D20+DCl (ppm):2.12(3H,s),3.20(3H,s),3.25, 3.50(2Hs~B-q,J-l~Hz),5.09 (lH,d9J=4.5Hz), 5.33(1H,s),5.66(1H,d~ J=
4.5Hz),7.15(1H,d,J=8Hz), 7.40(1H,d,d9J=2Hæ,8Hz), 7.53(1H,djJ=2Hz).

~ 20 : ' , . : ~ .: . . .
; ~ , ,., . : , . . - , .
. . .
~:.,. . ~: ,. .

Example 17. iO 4Z ~2 Z
(1) A solution of triethylamine(l40mg.) in methylene chloride (5 ml.) and 2,6-lutidine(15mg.) were added -to a solution of hydrochloride (540mg.) of 2,2,2-trichloroethyl 7-amino-3-methyl-3-cephem-4-carboxylate (540mg.) in dry methylene chloride - (10 ml.) under ice cooling. To the mixture was added a methylene chloride solution t5 ml.) containing N-tert-butoxycarbonyl-2-(3-ethylaminosulfonamidophenyl)-D-glycine (530 mg.) with stirring under ice cooling. A~ter N,N'-dicyclohexylcarbodiimide (320 mg.) was added thereto, the resultant mixture was stirred at room temperature for 3 hours. An insoluble product in the resultant mixture was ~iltered and washed with methylene chloride. The filtrate and the washings were combined together, and then the mixture was ~ -concentrated under reduced pressure. Ethyl aceta-te was added to the residue and an insoluble product was ~iltered of~. The ethyl acetate layer was washed with cold 5~ hydrochloric acid, water, an aqueous saturated solution o~ sodium bicarbonate and an aqueous saturated of sodium chloride, respectively, and then dried. Ethyl acetate was added to the solutlon and an insoluble product was ~iltered of~, and then the resultant solution was concentrated under reduced pressure to give an amorphous product (1.07g.) of 2,2,2 trichloroethyl 7-(N tert-butoxycarbonyl-2-(3-ethylaminosulfonamidophenyl)-D-glycinamido~-~o 3-methyl-3-cephem-4-carboxyla-te, m.p. 70 to 73C(dec.).

':,: . . : . ' . .

I.R. ~04Z422 Nujol V (cm~ ):3290,1770,1680 (2) Ace-tic acid (1.2 ml.) and zinc powder (0.96g.) were added under ice cooling to a solution o~
2,2,2-trichloroe-thyl-7-(N-tert-butoxycarbonyl-2-(3-ethylaminosulfonamidophenyl)-D-glycinamido~-3 -cephem-4-carboxylate (0.94g.) in dimethylformamide (5 ml.). The mixture was kept stirring at the same temperature for an hour. A~ter the reaction was over, zinc powder was filtered, washed with ~ ~
ethyl acetate and -the washings were combined ; ~ ~ ;
together with the ~iltrate.
The mixture was poured into cold 5% hydrochloric acid (20 ml.) and extracted twice with ethyl acetate, and then the extract was washed with an aqueous saturated solution o~ sodium chloride and back~
extracted with 5% aqueous solution of sodium bicarbonate. ~he aqueous layer was separated out, adjusted with 10% hydrochloric acid to pH 1-2 and extracted twice with ethyl acetate. The extract was washed with an aqueous saturated solution of sodium chloride~ dried and concentrated ~der reduced pressure to give an amorphous product (0.68g) of 7-[N-tert-butoxycarbonyl-2-(3-ethylaminosul~onamidophenyl)-D-glycinamido)-~-methyl-3-cephem-4-carboxylic acid, m.p. 122 to 128C ~dec.).

I.R.
Nujol (cm~l): 3300, 1765, 1690 ~, ,, .: - ,.~ . .

~ O~
N,M.R.
.~ C D2~NaHC
d (ppm) :1.03(3H~t)~1~40(gH~s)~
1.9~(3H,s),2.6-3.5(4H, m)J5.0(1H,d)5.30(1H, s)~5.60(1H,d),7.30(4H, m).

(3) A mixture of 7-(N-tert-butoxycarbonyl-2-(3-ethylaminosul~onamidophenyl)-D-glycinamidoJ-3-me-thyl-3-cephem-4-carboxylic acid t590 mg.) in formic acid (8 ml.) was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure and the residue was dis-solved in water. Ethyl acetate was added to ~ -the solution and the mixture was shaken. After ~ ;
the aqueous layer was separated out and concentrated, the resldue was adjusted to pH 5 with 5% aqueous solution o~ sodium hydroxide adsorbed on an adsorption resin ~XAE-2* produced by Rohm and Haas - Co.), washed with water and eluted with methanol.
The methanol elua-te was concentrated under reduced , pressure and the residue was washed with acetonitrile to give 7-[2-(3-ethylaminosul~onamidophenyl)-D-glycinamido]-3-methyl-3-cephem-4-carboxylic acid (350mg.), m.p. 162 to 166C.

I.R. ~
Nujol ':
(cm~l) : 3200, 1760, 1690 ' -* trademark . , , . ~ , ~ . , .

~04Z~2Z
N.M.R.

(ppm):1.03(3H,t),3.03(2~,q),3.0, 3.45(2H,d,d,J=19Hz),4.95 (lH,d),5.27(1H,broad s), 5.61(lH,d),7.30(4H,m).

Example 18.
(1) To a suspension o~ hydrochloride (3.61g.) of 2,2,2- .
trichloroethyl 7-amino-3-methyl-3-cephem-4-carboxylate in methylene chloride (50 ml.) was added triethylamine (0.95g.) all at once at 5C. N-tert-butoxycarbonyl-2- ~-(3-methylureido)phenyl~-D-glycine (3.37g.) and N, N'-dicyclohexylcarbodiimide (2.88g.) were successively added to the mixture and stirred at room temperature for an hour. An insoluble product was filtered off and the ~iltrate was concentrated under reduced pressure, and then ethyl acetate was added to the residue. The resultant solution was washed with 5% -hydrochloric acid (three times), water, an aqueous solution of sodium bicarbonate (three -times)~ water and an a~ueous solution of sodium chloride (three times), in turn, dried and then concentrated. The concentrate was allowed to stand, at a cold place and the appearing crystals were filtered -to give 2,2,2-trichloroethyl 7-[N-tert-butoxylcarbonyl-2-{3-(3-methylureido)phenyl}-D-glycinamido~
~-methyl-3-cephem-4-carboxylate (3.96g.), m.p.
152 to 160C (dec.).

.
., ~ . . ., ~ , , . . !

I.R. ~4~4ZZ
Nujol (cm~ ): 3320,1782,1731 N.M.R.
~DMS0-d6 (ppm): 1.42(9H,s),2.08(3H,s), 2.63(3H,d,J=5Hz),3.47(2H, t, J=20Hz),4.8-5.4(4H,m) 7 5.95(1H,d,J=5Hz),6.7-7.6 (4H,m),8.37(1H,s),9.05(1H, - -d ~ Ja7Hz ) ~

(2) Zinc powder (5.7g.) and glacial acetic acid ; (5.7 ml.) were added at 5C to a solution of 2,2,2-trichloroethyl 7-(N-tert-butoxycarbonyl-2-~3-(3-methylureido)phenyl~-D-glycinamido~-3-methyl-3-cephem-4-carboxylate (5.7g.) in ~ -dimethyl~ormamide (57 ml.). The mixture was stirred for an hour at room temperature and filtered. To the filtrate, was added a little o~ ice, 5% hydrochloric acid and ethyl acetate, The organic layer was separated out, washed twice with water and back-extracted with aqueous solution of sodium bicarbonate. The aqueous layer was separated out, extracted with ethyl acetate, acidi~ied with 5% hydrochloric l;.
acid and then extracted with ethyl acetate.
The extract was washe~ with wa-ter, dried over magnesium sulfate and then the solvent was removed from the extract to give 7-[N-tert-butoxycarbonyl-2-{3-~3-methylureido)phenyl~-., D-glycinamido~-3-methyl-3-cephern-4-carboxylic acid (3.2gg.), m.p. 143 to 150C (dec.).

I.R.

V (cm ) : 3370, 1780 N.M.R.
DMSO-d (ppm) : 1.38(9H,s),1.99(3H,s), 2.63(3H,d,J=5Hz),3.28, 3.45(2H,AB-q,J_18Hz), 4.96(1H,d,J=5Hz),5.28 (lH,broad d,J=8Hz),5.62 (lH,d,d,J=5Hz,8Hz), 5.97(1H,broadl d,J=5Hz), -6.8-7.6(4H,-m)9 8.44(1H,s), 9.02(1H,broad d,J=8Hz). ~-(3) 7-[N-tert-Butoxycarbonyl-2-{3-(3-methylureido)phenyl}-D-glycinamido~-3-methyl-3-cephem-4-carboxylic acid (3.18g.) was added to 20 ~ formic acid (60 mi.) at 10C,;and the mixture was stirred at room temperature for 2 hours, and concentrated under reduced pressure. The residue was pulverized with ether (70 ml.), and the filte~ed powder was added;to 5% aqueous acetonitrile (10 ml.), stirrecl for 30 minutes and then filtered to give - crystals (2.2g.) of 7-~Z-{~-~3-methylureido)-phenyl~ -D-glycinamido)-3-methyl_3-cephem-4-carboxylic acid~ m.p. 215 to 218C ~dec.).

E - 36 ,-?: ,-~ ~

I.R. 10~2422 NUJOl ~ (cm~l) : 3350, 1760 N.M.R.
D O+DC~
2 (ppm) : 2.2~3H,s),2.~5(3H,s), 3.23,3.51(2H,AB-q,J=l9 Hz),4.95(1H,d,J=5Hz), 5.33(1H,s),5.7(1H,d,J=
5Hz),7.2-7.7(4H,m).

Example 19.
(1) A solution of ethyl chlorocarbonate (1.79g.) in methylene chloride (10 ml.) was added to a solution of N-tert-butoxycarbonyl-2-{3-(3-methylthioureido) phenyl}-D-glycine (6.45g.) and triethylamine (1.82g.) in methylene chloride (75 ml.) at -25 to -30C in . .
10 minutes and the mixture was kept stirred at the same temperature for 15 minutes. A solution o~
2p2,2-trichloroethyl 7-amino-3-methyl-3-cephem-4-carboxylate hydrochloride (5.73g.) and triethylamine (1.52g.) in methylene chloride (75 ml.) was added to the above obtained mixture at -40C
all at once. The resultant mixture was gradually ;
warmed to 0C in 4 hours under stirring and - concen-trated under reduced pressure and the residue was dissolved in ethyl acetate (150 ml.). The solution was washed with 5% hydrochloric acid~ an aqueous saturated solution of sodium bicarbonate and aqueous saturated solution of .sodium chloride, E ~ 37 _ 50 _ lO~Z4~Z
respectively, and then the solvent was removed under reduced pressure. The residue was purified by column chromatography on 8ilica gel[eluant:
benzene (5 parts) + acetone (1 part)~ to give 2, 2,2-trichloroethyl 7~[N-tert-butoxycarbonyl-2-~3-(3-methylthioureido)phenyl~-D-glycinamido)-3-methyl-3-cephem-4-carboxylate (3.83g.), m.p.
123 to 130C.

I .R.
Nujol V (cm~l) : 3320, 1778, 1680, 1535, 1367, 1~06, 1243, 1217, 1162, 1110, 1050, 787, 718 .
N M.R.
~DMS0-d~
(ppm): 1.39(9H,s),2.08(3H,s),2.91(3H, d,J=4Hz),3.41(2H,broad s), 4.8-5.1(2H,m),5.28(1H, broad.
d,J=7Hz),5.5-5.8(1H,q)j7.0-7.75(5H,m),9.10(lH,broad d ~ ~ J=7Hz),9.48(1H,s).

;~ (Z) To a solution o~ 2,2,2-trichloroe-thyl 7-[N-tert~
-~ butoxycarbonyl-2-r3-(3-methylthioureido)phenyl~-D-glycinamido~-3-methyl-3-cephem-4-carboxylate ;~ 25 (3.55g.) and glacial acetic acid (3 ml.) in dimethylformamide (15 ml.), was added zinc powder (3.0g.), and the mixture was stirred under ice cooling for 1.5 hours. After the reaction was over, the zinc powder was filtered, and washed ., . .... ,, .. : . ., : . . : . . : .

ll~4~ZZ :
with ethyl acetate (50 ml.), and then the washings and the filtrate were combined together. After the mix-ture was washed with water, the aqueous layer was separated and extracted twice with ethyl acetate (30 ml.). The extract was combined -toge~her with the above obtained ethyl acetate layer. The ethyl acetat~ layer was washed with an aqueous saturated solution o~ sodium chloride and then back-extracted ~our times with an aqueous saturated solution of sodium bicarbonate (40 ml.).
The aqueous layer was washed with ethyl acetate, -adjusted to pH 2 with 10% hydrochloric acid and ~
then extracted ~ive times with ethyl acetate (60 ml.). ~ -The extr~ct was washed with water and dried over ~ ~
magnesium sulfate, and then the solvent was removed ~;
under reduced pressure to give a powdery product -~
(2.45g.) o~ 7-[N-tert-butoxycarbonyl-2-~3-(3- ` -;
methylthioureido)phenyl}-D-gl~cinamido~-3-methyl_ -~
3-cephem-4-carboxylic acld.

~ I.R.

(cm ): 3320, 1700, 1670, 1540, 1370 1250, 1165, 1110, 1056, 8667 718 N.M.R.
~DMSO-d6 (ppm):1.40(9H9s),2.00(3H, broad ~ :
s)~2.92(3H,s),3.1-3.7~2H, broad s~,4.99(1H,d9J=5Hz), 5.33(1H, broad d,J=8Hz~, 5.5-5.8(1H,m),7.0-7.7(4H, m),9.07(1H, broad d9J~8Hz), 9.50(1H, broad s).

ao~2~2 (3) A solution of 7-lN-tert-butoxycarbonyl-2- ~
methylthioureido)phenyl~--D-glycinamido]-3-methyl-3-cephem-4-carboxylic aci.d (2.1g.) in formic acid (20 ml.) was stirred at room temperature for 3 hours.
After the reaction was over, the formic acid was removed under reduced pressure and -the residue was stirred in 10% aqueous acetonitrile (20 ml.). The precipitates were filtered, washed with acetonitrile and dried to give 7-[2-{3-(3-methylthioureido)phenyl~-D-glycinamido~-3-methyl-3-cephem-4-carboxylic acid (1.21g.), m.p. 198 to 199C (dec.).

I R
Nujol ~J (cm~l) : 3500, 3430, 3230, 2600-2700, 1767, 1700, 1608, 1550, 141~, 1328~ 1280, 1230, 1178, 1160, 1130) 978, 8129 788, 757, 711 .~:
N~M.R.
CD20+DC~
~ (ppm) : 2.07(3H,s),3.02(3H,s), 3 34, 3 . 37 ( 2H, AB-q, Jal8 ~ '.
Hz),5.09(1H,d,J=4.5Hz), 5.37(1H,s),7.52(4H, broad ~ ~
: s ) . . ~, .

Example 20.
Sodium bicarbona-te (0.86g.) was added to the stirred suspension of 7-(D-N-tert-butoxycarbonyl-2-(3-mesylaminophenyl)-glycinamido~cephalosporanic acid (3g.) in pH 6.4 phosphate buffer (130 ml.) and then acetone (80 ml.) was added thereto. 5-Methyl-1,3, ! E - 40 ' '.'' ''' ~ . '.~' , ' ,. :'.,''",' .; ' ' ~Z4;~:Z
4-~hia~iazole-2-thiol (0.68g.) wais added to the solution and stirred a-l; 60 to 65C for 6 hours.
Ace-tone was removed ~rom the resultan-t mixture under reduced pressure and the remaining aqueous layer was washed with ether. The a~ueoUs solution was adjusted to pH 2 With diluted hydrochloric acid and then extracted with ethyl acetate.
The extract was washed with water, dried over magnesium sulfate and -then the solvent was remo~ed under reduced pressure. The residue was purified by column chromatography on silica gel (eluant:
chloroform) to give 7-[D-N-tert-butoxycarbonyl-2-(3-mesylaminophenyl)glycinamido]-3-(5-me~hyl-1,-3, ; 4-thiadiazol-2-yl)thiomethyl-3-cephem-4-carboxylic acid (1.8g.).

I~R.
Nujol (cm~l) : 3300, 1780, 1725, 1685, 1670 :~ ' ~, ~; 30 - -: - . : ~ :

Claims (19)

Tile embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A process for the preparation of a compound of the formula (I):
(I) wherein R1 is a nitro group or a lower alkoxy group, and a nontoxic, pharmaceutically acceptable salt or ester thereof, which comprises acylating 7-amino-3-methyl-3-cephem-4-carboxylic acid of the formula (II):
(II) or a derivative at the carboxy group and/or the amino group thereof, with a substituted phenylglycine of the formula (III):
(III) wherein R1 is as defined above and X is amino group or a pro-tected amino group, or a reactive derivative at the carboxy group, and then, if necessary, removing a protective group on the amino group and carboxy group, and, when desired converting a free acid of formula (I) into a corresponding nontoxic, pharmaceutically acceptable salt or ester thereof.

2. A process accordiny to claim 1, wherein the 7-amino-3-methyl-3-cephem-4-carboxylic acid of formula (II) or a tri-chloroethyl ester thereof is acylated with a mixed acid anhydride of the substituted phenylglycine of formula (III) with pivaloyl chloride.

3. A process according to claim 1, wherein X of the compound (III) is a lower alkoxycarbonylamino.

4. A process according to claim 1, wherein R1 is a nitro group.

5. A process according to claim 4, for producing 7-[D-2-(3-nitro-4-hydroxyphenyl)glycinamido]-3-methyl-3-cephem-4-carboxylic acid comprising acylating 7-amino-3-methyl-3-cephem-4-carboxylic acid or a trichloroethyl ester thereof, with N-t-butoxy-carbonyl-D-2-(3-nitro-4-hydroxyphenyl)glycine.

6. A process according to claim 1, wherein R1 is lower alkoxy.

7. A process according to claim 6, for preparing 7-[D-2-(3-methoxy-4-hydroxyphenyl)glycinamido]-3-methyl-3-cephem-4-carboxylic acid comprising acylating 7-amino-3-methyl-3-cephem-4-carboxylic acid or a trichloroethyl ester thereof, with N-t-butoxycarbonyl-2-(3-methoxy-4-hydroxyphenyl)glycine.

8. A process according to claim 1, including the step of reacting a free acid of formula (I) with a nontoxic inorganic or organic base to produce a corresponding nontoxic, pharmaceutically acceptable salt.

9. A process according to claim 6, wherein R1 is lower alkoxy of 1 to 6 carbon atoms.

10. A process according to claim 1, 4 or 9, wherein said acylating is carried out in an inert organic solvent.

11. A process according to claim 1, 4 or 9, wherein said acylating is carried out in the presence of a base.

12. A process according to claim 1, 4 or 9, wherein said substituted phenylglycine is in the form of the free acid or a salt thereof, and said acylating is carried out in the presence of a condensing agent.

13. A compound of the formula:- (I) wherein R1 is a nitro group or a lower alkoxy group, and a nontoxic, pharmaceutically acceptable salt or ester thereof, whenever prepared by the process of claim 1, or by an obvious chemical equivalent thereof.

14. A compound of the formula (I) as defined in claim 1, wherein R1 is a nitro group whenever prepared by the process of claim 4, or by an obvious chemical equivalent thereof.

15. A compound of formula (I) as defined in claim 1, wherein R1 is lower alkoxy whenever prepared by the process of claim 6, or by an obvious chemical equivalent thereof.

16. A compound of formula (I), as clefined in claim 1, wherein R1 is lower alkoxy of 1 to 6 carbon atoms, whenever prepared by the process of claim 9, or by an obvious chemical equivalent thereof.

17. A nontoxic, pharmaceutically acceptable salt of a compound of formula (I), as defined in claim 1, whenever pre-pared by the process of claim 8, or by an obvious chemical equivalent thereof.

18. 7-[D-2-(3-nitro-4-hydroxyphenyl)glycinamido]-3-methyl-3-cephem-4-carboxylic acid, whenever prepared by the process of claim 5, or by an obvious chemical equivalent thereof.

19. 7-[D-2-(3-methoxy-4-hydroxyphenyl)glycinamido]-3-methyl-3-cephem-4-carboxylic acid, whenever prepared by the process of claim 7, or by an obvious chemical equivalent thereof.