Novel 3-acyloxymethyl-cephem compounds of the formula: ##STR1## wherein R.sup.1 is hydrogen or an acyl group; X is a divalent group consisting of a carbon chain having 2 to 3 carbon atoms and a carbonyl or sulfonyl group at one terminal end thereof, said divalent group being either substituted or unsubstituted on the carbon chain; and Z is an organic acid residue, and salts thereof were found to be useful as starting materials for preparing cephalosporins of the formula: ##STR2## wherein R.sup.2 stands for a residue of a nucleophilic compound and R.sup.1 has the same meaning as above.

The present invention relates to novel 3-acyloxymethyl-cephem compounds and 
preparations thereof. More particularly, this invention relates to the 
compounds of the formula (I); 
##STR3## 
wherein R.sup.1 is hydrogen or an acyl group; X is a divalent group 
consisting of a carbon chain having 2 to 3 carbon atoms and a carbonyl or 
sulfonyl group at one terminal end thereof, said divalent group being 
either substituted or unsubstituted on the carbon chain; and Z is an 
organic acid residue, and pharmaceutically acceptable salts thereof, and 
also relates to processes for producing them. 
Cephalosporin derivatives with a 3-hydroxymethyl moiety were only 
obtainable by enzymatic cleavage of the 3-acetyl group from 
3-acetoxymethyl-cephalosporins or by separating them from the 
cephalosporin C fermentation by-product. Recently, it has become possible 
to produce 
7.beta.-(D-5-amino-5-carboxyvaleramido)-3-hydroxymethyl-3-cephem-4-carboxy 
lic acid (deacetylcephalosporin C, DCPC) in high titer by fermentation 
(U.S. Pat. No. 3,926,726, Nature New Biology, 246, 154(1973)) and, 
alongside of cephalosporin C, this substance has been attracting attention 
as a starting material for cephalosporin compounds that could be more 
potent in antibiotic activity. 
It has, however, been believed to be difficult to acylate the 
3-hydroxymethyl group of the 3-hydroxymethyl compound (cephalosporadesic 
acid). For example, Heyningen [Van Heyningen: J. Med. Chem., 8, 22(1965), 
Advan. Drug. Res., 4, 28(1968)] reported that the O-acylation of 
cephalosporadesic acid was feasible only with the use of a large excess of 
aroyl chloride (yield 32%-57%) and that the use of ketene, aliphatic acid 
chloride or acetic anhydride did not cause the O-acylation, or induced a 
lactonization. Kukolja [J. Med. Chem., 13, 1114(1970)] reported a 
roundabout process for the synthesis of O-acyloxymethylcephalosporins 
which comprised O-acylating a 3-hydroxymethyl-2-cephem compound and then 
causing the latter to become isomerized to the 3-cephem compound. U.S. 
Pat. No. 3,532,694 and Japanese Patent Publication No. 33080/1975 
disclosed a process in which, to prevent the lactonization reaction, the 
4-carboxyl group of cephalosporadesic acid is first protected, for example 
by esterification and, then, the O-acylation is carried out. Disclosed in 
Japanese Patent Application as Laid Open No. 42792/1972 is a process which 
comprises O-acylating cephalosporadesic acid with azolide. However, these 
processes are not commercially profitable because they provide only low 
yields or/and involve troublesome and time-comsuming procedures or/and 
expensive reagents, for instance. Thus, for example, the esterification 
reaction of cephalosporadesic acid cannot be accomplished by an ordinary 
esterification process in which the rearrangement of the double bond or 
the lactonization predominates. While it is possible to introduce such 
limited groups as, e.g., methyl, ethyl, diphenylmethyl or benzyl by means 
of diazo compounds which as diazomethane, diazoethane, 
diphenyldiazomethane, phenyldiazomethane, for example, it is difficult, 
after 3-acylation, to de-esterify the compound without the accompaniment 
of some side reaction such as a fission of the .beta.-lactam ring or a 
shift of the double bond. 
On the other hand, the reaction by which the 3-acetoxymethyl group of a 
cephalosporin compound is substituted with a nucleophilic reagent entails 
a concurrent decomposition of the starting material, intermediate and 
product in its course and a protracted reaction time, and therefore 
results in lower yields [A. B. Tayor, J. Chem. Soc., 7020(1965)]. Thus, it 
has been desired to have available a derivative possessing a group which 
will lend itself more readily to substitution than the acetoxy group. 
To overcome the foregoing problems we undertook extensive research, which 
led us to the finding that the use of compounds [IV] hereinafter described 
as an acylating agent would enable cephalosporadesic acid to be O-acylated 
in a high yield, and that the O-acylated cephalosporin thus synthesized 
would undergo the desired substitution with a nucleophilic compound with 
great ease. This invention has been developed on the basis of the above 
findings. 
The above compounds [I] include the compounds wherein R.sup.1 is hydrogen, 
phenylacetyl, phenoxyacetyl, 5-amino-5-carboxyvaleryl whose amino or/and 
carboxyl groups may optionally be protected, or any of the groups found in 
the 6- or 7-positions of penicillin or cephalosporin derivatives, as the 
case may be. Thus, for example, the acyl group R.sup.1 may be chosen from 
among aliphatic acyl groups such as formyl, acetyl, propionyl, butyryl, 
isobutyryl, valeryl, isovaleryl, cyclopentylcarbonyl, cyclohexylcarbonyl, 
cycloheptylcarbonyl, cyclopentylacetyl, cyclohexadienylacetyl, etc.; 
aromatic acyl groups such as benzoyl, p-nitrobenzoyl, toluoyl, naphthoyl, 
etc.; mono-substituted aliphatic acyl groups such as 2-thienylacetyl, 
cyanoacetyl, acetoacetyl, 4-chloro-3-oxobutyryl, 4-bromo-3-oxobutyryl, 
chloroacetyl, bromoacetyl, 4-methylthio-3-oxobutyryl, 
4-carbamoylmethylthio-3-oxobutyryl, .alpha.-phenoxypropionyl, 
.alpha.-phenoxybutyryl, tetrazolylthioacetyl, tetrazolylacetyl, 
p-nitrophenylacetyl, trifluoromethylthioacetyl, 
trifluoromethylsulfinylacetyl, trifluoromethylsulfonylacetyl, 
cyanomethylthioacetyl, thiadiazolylthioacetyl, p-nitrophenylacetyl, 
(2-pyridyloxy)acetyl, (2-oxo-4-thiazolin-4-yl)acetyl, 
(2-imino-4-thiazolin-4-yl)acetyl, (2-thioxo-4-thiazolin-4-yl)acetyl, 
4-pyridylthioacetyl, (3-sydnone)acetyl, 1-pyrazolylacetyl, 2-furylacetyl, 
(2-oxo-3-methylpyridazinyl)thioacetyl, (2-aminomethylphenyl)acetyl, 
(2-aminomethylcyclohexenyl)acetyl, etc.; di-substituted aliphatic acyl 
groups such as .alpha.-carboxyphenylacetyl, mandelyl, 
.alpha.-sulfophenylacetyl, .alpha.-sulfo-(p-aminophenyl)acetyl, 
phenylglycyl, (4-hydroxyphenyl)glycyl, (4-methylthiophenyl)glycyl, 
(4-methoxyphenyl)glycyl, (4-methanesulfinylphenyl)glycyl, 
(3-methanesulfonamidophenyl)glycyl, 1-cyclohexenylglycyl, thienylglycyl, 
furylglycyl, cyclohexadienylglycyl, (3,4-dihydroxyphenyl)glycyl, etc.; 
5-methyl-3-phenyl-4-isoxazolylcarbonyl; 
3-(2,6-dichlorophenyl)-5-methyl-4-isoxazolylcarbonyl; and so forth. It 
should be understood that the above-mentioned groups are only illustrative 
of the acyl groups that are capable of use for the purposes of this 
invention, but preferred acyl groups may be represented by the formula 
(A): 
##STR4## 
wherein R.sup.4 represents a group such as, for example, acetyl, 
halogenoacetyl, phenyl, p-hydroxyphenyl, thienyl, 
2-imino-4-thiazolin-4-yl, 2-oxo-4-thiazolin-4-yl, tetrazolyl, phenoxy, 
3-amino-3-carboxypropyl, and R.sup.5 represents a substituent such as, for 
example, hydrogen, sulfo, amino or hydroxy, etc. 
It should also be understood that any functional groups, e.g. amino or/and 
carboxyl, in such acyl groups may be suitably protected by conventional 
protective groups. Thus, among such protective groups for the amino groups 
are aromatic acyl groups such as phthaloyl, benzoyl, p-nitrobenzoyl, 
toluoyl, naphthoyl, p-tert-butylbenzoyl, p-tert-butylbenzenesulfonyl, 
phenylacetyl, benzenesulfonyl, phenoxyacetyl, toluenesulfonyl, 
chlorobenzoyl, etc.; aliphatic acyl groups such as acetyl, valeryl, 
capryl, n-decanoyl, acryloyl, pivaloyl, camphorsulfonyl, methanesulfonyl, 
chloroacetyl, etc.; esterified carboxyl groups such as 
tert-butoxycarbonyl, ethoxycarbonyl, isobornyloxycarbonyl, 
phenyloxycarbonyl, trichloroethoxycarbonyl, benzyloxycarbonyl, 
.beta.-methylsulfonylethoxycarbonyl, etc.; carbamoyl groups such as 
methylcarbamoyl, phenylcarbamoyl, naphthylcarbamoyl, etc.; the 
corresponding thiocarbamoyl groups; 2-methoxycarbonyl-1-methylvinyl; and 
so forth. As protective groups for the carboxyl groups of such acyls 
R.sup.1 and the 4-carboxyl group of the cephem ring, there may be 
mentioned methyl, ethyl, tert-butyl, tert-amyl, benzyl, p-nitrobenzyl, 
p-methoxybenzyl, benzhydryl, 1-indanyl, phenacyl, phenyl, p-nitrophenyl, 
methoxymethyl, ethoxymethyl, benzyloxymethyl, acetoxymethyl, 
pivaloyloxymethyl, .beta.-methylsulfonylethyl, methylthiomethyl, trityl, 
.beta.,.beta.,.beta.-trichloroethyl, silyl groups such as trimethylsilyl, 
dimetlylsilyl, etc.; and so forth. These carboxyl groups, however, may be 
preferably in the form of inorganic or organic salts with alkali metals 
such as lithium, sodium, potassium, etc.; alkaline earth metals such as 
calcium, magnesium, or various amines such as dicyclohexylamine, 
triethylamine, tributylamine, di-n-butylamine, di-n-propylamine and so 
forth. 
The divalent group X means a hydrocarbon group which has a carbonyl or 
sulfonyl group at one of its terminal ends and which is normally able to 
form a five-membered or six-membered ring with 
##STR5## 
and may be interrupted by, or include at one terminal end thereof, an 
unsaturation such as a double bond or oxygen or/and sulfur atoms. The 
hydrocarbon chain may further carry suitable substituents. As examples of 
such substituents on said hydrocarbon chain, there may be mentioned alkyl 
groups such as methyl, ethyl, propyl, etc.; aralkyl groups such as benzyl, 
phenethyl, etc.; and aryl groups such as phenyl, tolyl, etc. Where two or 
more such substituents are present, they may form a cyclic structure with 
the hydrocarbon chain. The symbol Z is a group exemplified by lower 
alkoxycarbonyl groups such as methoxycarbonyl, ethoxycarbonyl, etc.; acyl 
groups such as acetyl, propionyl, benzoyl, chloroacetyl, etc.; other 
organic acid residues, e.g. sulfonyl derivatives such as phenylsulfonyl, 
tosyl, mesyl, etc.; and carbamoyl groups; phosphorous derivatives such as 
diethylphosphoro, dimethylphosphono, diethylphosphino, dimethylphosphino, 
and so forth. 
Those compounds [I] can be produced by reacting a compound of the formula: 
##STR6## 
wherein R.sup.3 is hydrogen or an acyl group, with a compound of the 
general formula: 
##STR7## 
wherein the various symbols are as defined above. 
Thus, compound [IV] is exemplified by N-carboethoxyphthalimide, 
N-carbomethoxypathalimide, 4-nitro-N-carboethoxyphthalimide, 
3-nitro-N-carboethoxyphthalimide, N-carboethoxysuccinimide, 
N-carbomethoxysuccinimide, N-tosylphthalimide, 
N-methylsulfonylphthalimide, N-benzenesulfonylphthalimide, 
N-acetylphthalimide, N-chloroacetylphthalimide, N-acetylsuccinimide, 
N-carboethoxysaccharin, N-acetylsaccharin, N-benzoylsaccharin, 
N-carboethoxymaleimide, N-carboethoxyglutarimide, 
N-carboethoxy-(p-chlorophenyl)succinimide, N-carboethoxyisatin, 
N-carbomethoxyisatin, N-acetylisatin, N-(methylcarbamoyl)isatin, 
N-(phenylcarbamoyl)isatin, N-(.beta.-methylsulfonylethoxycarbonyl)isatin, 
N-(diethylphosphoro)succinimide, N-(dimethylphosphoro)succinimide, 
N-(dimethylphosphino)succinimide, N-(diethylphosphino)phthalimide and so 
forth. 
Generally the reaction of compound [II] with compound [IV] is conveniently 
carried out in the presence of an appropriate inert solvent. Among the 
common solvents used in this reaction are dichloromethane, chloroform, 
dichloroethane, ethyl acetate, acetonitrile, acetone, tetrahydrofuran, 
dimethylformamide, dimethylacetamide, dioxane, ether and mixtures of such 
solvents. This reaction is a stoichiometrically equivalent reaction, that 
is to say, [II] and [IV] may be reacted in equimolar proportions, although 
an excess of [IV] may be used to hasten the reaction, to compensate for 
the decomposition of [IV] or for other considerations. Generally the 
reaction may be conducted at room temperature or under cooling (for 
example, -10.degree. C. to -40.degree. C.), preferably at 0.degree. to 
30.degree. C. The reaction normally goes to completion within a short 
time, but in view of the nature of the reaction, which is somewhat 
temperature-dependent, the reaction is usually carried out for 0.5 to 15 
hours so as to ensure a thorough completion of the reaction. If necessary, 
an amine exemplified by triethylamine may be added to the reaction system. 
Where the starting compounds [II] is an alkali metal salt, a salt 
interchange may be effected by the addition of an equivalent of 
triethylamine hydrochloride or the like before the said reaction is 
conducted. 
Where starting compounds [II] contain a free, unprotected amino group, this 
amino group is acylated simulteneously with the acylation of the 
3-hydroxyl group by the same acyl group under the conditions of this 
reaction. In other words, the protective group for amino in the acyl group 
includes the acyl groups formed on reaction with [IV] in addition to the 
groups mentioned hereinbefore. The N-acylated compound [I] or [II] wherein 
R.sup.1 or R.sup.3 is an acyl group respectively, when necessary, may be 
converted to the corresponding compound wherein R.sup.1 is hydrogen by 
cleaving the N-acyl group off in a manner conventional per se. The meaning 
of R.sup.3 only differs from that of R.sup.1 in these cases. 
The resultant compound [I] not only has antibiotic activity as such but is 
ready to react with a nucleophilic compound to have the residue of said 
nucleophilic compound introduced into the 3-methyl group of the 
cephalosporin to give the compound represented by the formula [III]: 
##STR8## 
wherein R.sup.2 stands for a residue of a nucleophilic compound and 
R.sup.1 has the same meaning as defined before. 
As to the nucleophilic compound used for this reaction, any compound able 
to replace the 3-acetoxy groups of cephalosporins may be used. 
Furthermore, this reaction proceeds far faster than the reaction of the 
3-acetoxy compound, with an additional advantage that a substituent group 
which does react with the 3-acetoxy compound, which is less active, only 
in poor yields can be successfully introduced. 
Therefore, among such nucleophilic compounds are nitrogen-containing 
heterocyclic thiols which contain one or more nitrogen atoms which may 
optionally be in the form of oxide or/and which contain such atoms as 
oxygen or/and sulfur in addition to the nitrogen atom, with or without 
nuclear substitution. As common examples of the nitrogen-containing 
heterocyclic group of such a thiol compound, there may be mentioned 
pyridyl, N-oxidepyridyl, pyrimidyl, pyridazinyl, N-oxidepyridazinyl, 
pyrazolyl, diazolyl, thiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 
1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,2,3-oxadiazolyl, 
1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,2,3-triazolyl, 
1,2,4-triazolyl, 1H-tetrazolyl, 2H-tetrazolyl and so on. As the 
substituents on such nitrogen-containing heterocyclic groups, there may be 
mentioned such monovalent groups as hydroxy, mercapto, amino, carboxyl, 
carbamoyl, lower alkyl (for example, methyl, ethyl, trifluoromethyl, 
propyl, isopropyl, butyl or isobutyl), lower alkoxy (for example, methoxy, 
ethoxy, propoxy, isopropoxy or butoxy), halogens (e.g. chlorine or 
bromine), and various substituent groups attached through lower alkylene 
groups, --S--, --N-- or other polyvalent groups. When such polyvalent 
groups are lower alkylene groups, the substituents may, for example, be 
hydroxy, mercapto, amino, morpholino, carboxyl, sulfo, carbamoyl, 
alkoxycarbonyl, lower alkylcarbamoyl, alkoxy, alkylthio, alkylsulfonyl, 
acyloxy, morpholinocarbonyl and so on. When such a polyvalent group is 
--S-- or --N--, the substituents may be lower alkyls or lower alkylene 
groups having the aforementioned substituents. When the polyvalent group 
is --N--, such substituents as carboxyl, alkoxycarbonyl, acyl, carbamoyl 
or lower alkylcarbamoyl may be directly attached. More particularly, there 
may be mentioned, for example, substituted alkyl groups such as 
carboxymethyl, carbamoylmethyl, N-lower alkylcarbamoylmethyl (e.g. 
N,N-dimethylcarbamoylmethyl), hydroxy lower alkyl (e.g. hydroxymethyl or 
2-hydroxyethyl), acyloxy lower alkyl (e.g. acetoxymethyl or 
2-acetoxyethyl), alkoxycarbonylmethyl (e.g. methoxycarbonylmethyl, 
hexyloxycarbonylmethyl or octyloxycarbonylmethyl), methylthiomethyl, 
methylsulfonylmethyl, N-lower alkylamino lower alkyl (e.g. 
N,N-dimethylaminomethyl, N,N-dimethylaminoethyl or 
N,N,N-trimethylammoniumethyl), morpholinomethyl; substituted amino groups 
such as lower alkylamino (e.g. methylamino), sulfo-lower alkylamino (e.g. 
2-sulfoethylamino), hydroxylower alkylamino (e.g. hydroxyethylamino), 
lower alkylamino-lower alkylamino (e.g. 2-dimethylaminoethylamino or 
2-trimethylammoniumethylamino), acylamino (e.g. acetylamino), 
2-dimethylaminoacetylamino, 2-trimethylammoniumacetylamino or lower 
alkoxycarbonylamino (e.g. methoxycarbonylamino), etc.; and substituted 
thio (mercapto) groups such as methylthio, 2-hydroxyethylthio, 
2-acyloxyethylthio (e.g. 2-acetoxyethylthio, 2-phenylacetoxyethylthio or 
2-caproyloxyethylthio), carboxymethylthio, alkoxycarbonylmethylthio (e.g. 
methoxycarbonylthio or hexyloxycarbonylmethylthio), carbamoylmethylthio, 
N-lower alkylcarbamoylmethylthio (e.g. N,N-dimethylcarbamoylmethylthio), 
acetylmethylthio, N-lower alkylamino-lower alkylthio (e.g. 
2-N,N-dimethylaminoethylthio or 2-N,N,N-trimethylammoniumethylthio), 
morpholinocarbonylmethylthio, 2-sulfoethylthio and so on. More 
particularly, there may be mentioned various heterocyclic thiols such as 
tetrazolethiol, methyltetrazolethiol, phenyltetrazolethiol, 
(2-N,N-dimethylaminoethyl)tetrazolethiol, methylthiadiazolethiol, 
hydroxyethylthiothiadiazolethiol, methylthiothiadiazolethiol, 
thiadiazolethiol, carbamoylaminothiadiazolethiol, 
carbamoylmethylthiothiadiazolethiol, thiazolethiol, methylthiazolethiol, 
carboxymethylthiazolethiol, triazolethiol, dimethyltriazolethiol, 
pyrazolethiol, ethoxycarbonylmethyltriazolethiol, imidazolethiol, 
methyloxadiazolethiol, pyridinethiol, pyrimidinethiol, 
methylpyridazinethiol, triazinethiol and so on. In addition, use may also 
be made of such nitrogen-containing heterocyclic compounds as aliphatic or 
aromatic thiols, e.g. methanethiol, ethanethiol or thiophenol; thiourea 
and its derivatives such as N-methylthiourea or 
N-methyl-N'-pyridylthiourea; thioamide derivatives such as 
thiosemicarbazide, thioacetamide or thiobenzamide; sodium thiosulfate, 
sodium sulfite, potassium thiocyanate or sodium azide; pyridine and 
pyridine derivatives such as quinoline, picoline, nicotinic acid, 
nicotinamide, isonicotinamide, isonicotinic acid hydrazide, 
m-bromopyridine, pyridinesulfonic acid, pyridine-m-carbinol 
(3-hydroxymethylpyridine), pyridinaldehyde, quinoline or isoquinoline, 
etc.; and such other nitrogen-containing heterocyclic compounds as 
pyrazine, pyrazinamide (2-carbamoylpyrazine), pyridazine, pyrimidine, 
imidazole, 1-methylimidazole, pyrazol and so forth. It is also possible to 
employ carbon nucleophilic agents toward which the 3-position is known to 
be refractory. As examples of such carbon nucleophilic reagents, there may 
be mentioned cyanides, pyrrole, substituted pyrrole, indole, acetylene, 
active methylene compounds, e.g. acetylacetone, acetoacetic acid esters, 
malonic acid esters, cyclohexane-1,3-dione, triacetylmethane or enamine 
compounds. Alcohols such as, for example, methanol, ethanol or propanol 
may also be employed in this reaction. 
The substitution reaction between such a nucleophilic compound and a 
compound of formula [I] is normally conducted in a solvent. As said 
solvent, water may be mentioned as the commonest solvent, although a 
mixture of water and a reaction-inert hydrophilic solvent, e.g. acetone, 
acetonitrile, tetrahydrofuran, dimethylformamide, methanol, ethanol, 
dimethylsulfoxide or the like, may likewise be employed. The reaction may 
also be conducted even in a non-aqueous system. While compound [I] may be 
a free compound, it is more appropriate to react [I] in the form of an 
alkali metal salt, e.g. the salt of sodium, potassium or the like, or an 
organic amine salt, e.g. the salt of triethylamine, trimethylamine or the 
like. The nucleophilic compound, too, is reacted either in the free form 
or as an alkali metal salt or organic amine salt. The proper amount of 
said nucleophilic compound used in the reaction is not less than one 
equivalent based on compound [I]. While it depends upon the types of 
nucleophilic compound and compound [I], this reaction is preferably 
conducted generally under weakly acid to weakly alkaline conditions in 
cases where an aqueous solvent is employed. While the reaction temperature 
depends largely upon the nature of compound [I] and cannot be specified in 
general terms, it is preferably within the range of 10.degree. C. to 
70.degree. C. It is also possible to accomplish a direct synthesis of 
[III] by causing the nucleophilic reagent to be present concomitantly in 
the reaction system of [II] and IV]. The reaction time cannot be specified 
in general terms, either, for it depends upon the reaction temperature, 
pH, type of nucleophilic reagent and other factors. Roughly speaking, 
however, the reaction goes to completion in 30 minutes to 2 hours when the 
reaction temperature is 60.degree. C. The reaction may also be carried 
out in the presence of, as added to the reaction system, an inorganic 
salt, e.g. the chloride, bromide, iodide, thiocyanide or nitrate of 
lithium, sodium, potassium, ammonium or the like. At any event, the 
feasibility of the nucleophilic substitution reaction at such a low 
temperature and in a non-aqueous phase prevents decomposition of compound 
[I] and permits synthesis of the compound which cannot be synthesized in 
an aqueous phase. Where [I] has a high activity such that it will be 
readily substituted by a nucleophilic reagent at room temperature, for 
example in the case of an isatin derivative, [I] need not be isolated, but 
[III] may be directly synthesized by allowing a nucleophilic compound to 
be present in the reaction system when [II] is reacted with [IV]. 
The compound [I] wherein R.sup.1 is an acyl group can be converted to the 
corresponding compound wherein R.sup.1 is hydrogen by cleaving the 7-acyl 
group off in a manner conventional per se (e.g any of the procedures set 
forth in Japanese Patent Publication No.13862/1966 and No.40899/1970, 
Japanese Patent Application As Laid Open No.34387/1972, No. 95292/1975 and 
No.96591/1975, Japanese Patent Publication No.35079/1975 or United States 
Patent No. 3632578). 
Into this compound may be introduced a compound which has previously been 
mentioned by way of example for R.sup.1 as the 6- or 7-substituents of 
penicillin or cephalosporin compounds, by previously activating the same 
in a known manner. Thus, for example, the compound wherein R.sup.1 is 
4-halogeno-3-oxobutyryl can be obtained by reacting the said compound with 
a 4-halogeno-3-oxobutyryl halide which, in turn, may be reacted with 
thiourea to produce the 
7-[2-(2-imino-4-thiazolin-4-yl)acetamido]-compound. While it depends 
somewhat on the type of 3-substituent, these compounds invariably display 
excellent antibiotic activity. For example, the compound with 
1-methyltetrazol-5-ylthiomethyl in 3-position is particularly useful, 
approximately the same effect being accomplished with this compound at a 
dose about one-fifth that of cefazolin. 
The present invention is illustrated in further detail below with reference 
to examples, but it is to be understood that the examples are solely for 
the purpose of illustration and not to be construed as limitations of the 
invention, and that many variations may be resorted to without departing 
from the spirit and scope of the invention. In this specification, "g", 
"mg", "ml", "cm", "Hz", "DMSO", "arom" and "decomp." are abbreviations of 
"gram", "milligram", "milliliter", "centimeter", "Herz", 
"dimethylsulfoxide", "aromatic" and "decomposed", respectively. Resins 
named "Amberlite" are products manufactured by Rohm & Haas Co. in U.S.A. 
"Celite" and "Sephadex" are marketed by Johns-Manville Sales Corp. and 
Pharmacia A.B., respectively. All the temperatures are uncorrected and the 
percentages are all on the weight basis except those specifically defined. 
The NMR spectra were measured using a Varian Model HA 100 (100 MHz) or T60 
(60 MHz) spectrometer with tetramethylsilane as the internal reference 
and all .delta. values are in ppm. The symbols "s" stands for a singlet, 
"d" a doublet, "t" a triplet, "q" a quarter; "m" a multiplet, and "J" a 
coupling constant.

EXAMPLE 1 
In N,N-dimethylformamide (80 ml) is suspended deacetylcephalosporin C 
sodium monohydrate (purity 90%, 20 g) and, with the addition of 
concentrated hydrochloric acid (8 ml), the suspension is stirred under 
ice-cooling for 15 minutes. To this solution is added 
N,N-dimethylformamide (80 ml), followed by addition of 
N-carboethoxyphthalimide (40 g) and triethylamine (40 ml) in the order 
mentioned. The mixture is stirred at room temperature for 3 hours. While 
stirring it under ice-cooling, this reaction mixture is poured in 0.42% 
hydrochloric acid (2.0 l), followed by the addition of sodium chloride 
(380 g). The mixture is stirred for 30 minutes. The resultant precipitate 
is recovered by filtration, rinsed with a saturated aqueous solution of 
sodium chloride and dried over phosphorus pentoxide under reduced 
pressure. The product is suspended in ethyl acetate (1.2 l) and stirred at 
room temperature for 30 minutes. The insoluble fraction is removed by 
filtration and, under stirring, toluene (500 ml) is gently added. The 
mixture is concentrated under reduced pressure (to 500 ml) and the 
precipitate is recovered by filtration, rinsed with toluene and dried 
under reduced pressure. The procedure provides 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-[2-(N-carboethoxycarbamoyl 
)benzoyloxy]methyl-3-cephem-4-carboxylic acid (31 g). 
IR(KBr): cm.sup.-1 1771, 17155 
NMR(d.sub.6 -DMSO): .delta. 1.17(3H, t, J=7 Hz, OCH.sub.2 CH.sub.3), 
1.2-2.4(6H, m- --(CH.sub.2).sub.3 --), 3.56(2H, ABq, J=18 Hz, 
2--CH.sub.2), 4.05(2H, q, J=7 Hz, --OCH.sub.2 CH.sub.3), 4.74(1H, t, J=7 
Hz, 
##STR9## 
5.07(1H, d, J=5 Hz, 6-H), 5.08(2H, ABq, J=13 Hz, 3--CH.sub.2 O--), 
5.65(1H, q, J=5 & 8 Hz, 7-H), 7.36-7.94(8H, m, arom-H), 8.80(1H, d, J=8 
Hz, 7---CONH--), 11.08(1H, --CONH--CO--) 
EXAMPLE 2 
In cold water (100 ml) is suspended 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-[2-(N-carboethoxycarbamoyl 
)benzoyloxy]methyl-3-cephem-4-carboxylic acid (16.5 g), followed by the 
addition of 5-mercapto-1-methyl-1H-tetrazole (4.2 g). Then, under 
ice-cooling and stirring, sodium hydrogen carbonate (6.0 g) is added in 
small installments. To the resultant clear solution is added a further 
small amount of sodium hydrogen carbonate and, after adjustment to pH 5.0, 
sodium chloride (30 g) is added. The mixture is heated at 60.degree. C. 
for 1.5 hours. To this reaction mixture is added water (100 ml) and, under 
stirring, the mixture is adjusted to pH 1.5 with dilute hydrochloric acid. 
The resultant precipitate is recovered by filtration, rinsed with a 
saturated aqueous solution of sodium chloride and dried over phosphorus 
pentoxide under reduced pressure. The procedure provides 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-(1-methyl-1H-tetrazol-5-yl 
)thiomethyl-3-cephem-4-carboxylic acid (13.0 g). 
IR(KBr): cm.sup.-1 3325, 1780, 1730, 1715, 1650, 1545 
NMR(d.sub.6 -DMSO): .delta. 1.40-1.76(2H, m), 2.0-2.4(4H, m), 3.64(2H, ABq, 
J=19 Hz, 2--CH.sub.2), 3.93(3H, s, --NCH.sub.3), 4.30(2H, ABq, J=15 Hz, 
3--CH.sub.2), 4.73(1H, t, J=8 Hz, 
##STR10## 
5.01(1H, d, J=5 Hz, 6-H), 5.62(1H, dd, J--5 & 9 Hz, 7-H), 7.85(4H, s, 
arom-H), 8.80(1H, d, J=9 Hz, --CONH) 
EXAMPLE 3 
In N,N-dimethylformamide (0.4 l) is suspended deacetylcephalosporin C 
sodium monohydrate (purity 90%, 100 g) and, with the addition of 
concentrated hydrochloric acid (0.04 l), the suspension is stirred for 30 
minutes. To this reaction mixture is added N,N-dimethylformamide (0.3 l) 
and, then, N-carboethoxyphthalimide (200 g) and triethylamine (0.2 l) are 
added in the order mentioned. The mixture is stirred at room temperature 
for 3 hours. This reaction mixture is poured in an ice-cooled mixture of 
ethyl acetate (1.5 l), concentrated hydrochloric acid (120 ml) and 14% 
aqueous sodium chloride (2.0 l), followed by stirring for 20 minutes. The 
ethyl acetate layer is taken and the aqueous layer is extracted with ethyl 
acetate. The ethyl acetate extracts are pooled and rinsed with 14% aqueous 
sodium chloride. To this ethyl acetate solution is gently added a 4.75% 
aqueous solution of sodium hydrogen carbonate (10 l), followed by stirring 
for 15 minutes. The aqueous layer is taken and, following addition of 
sodium chloride (300 g) and with stirring, a mixture of 
5-mercapto-1-methyl-1H-tetrazole (32 g), sodium hydrogen carbonate (17 g) 
and water (0.12 l) is added. Then, the mixture is adjusted to pH 4.7 with 
a small amount of sodium hydrogen carbonate, followed by stirring at 
60.degree. C. for 1.5 hours. From the reaction mixture, the ethyl acetate 
is distilled off under reduced pressure. Following the addition of water 
(2.5 l), the mixture is adjusted to pH 1.5 with diluted hydrochloric acid 
and the resultant precipitate is recovered by filtration, rinsed with a 
saturated aqueous solution of sodium chloride and dried over phosphorus 
pentoxide under reduced pressure. The procedure provides 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-(1-methyl-1H-tetrazol-5-yl 
)thiomethyl-3-cephem-4-carboxylic acid (139 g). In IR and NMR spectra, this 
product is in good agreement with the product obtained in Example 2. 
EXAMPLE 4 
In N,N-dimethylformamide (40 ml) is dissolved 
7.beta.-(2-thienylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic acid 
sodium salt (3.8g), and following the addition of N-carboethoxysaccharin 
(4.0 g) and triethylamine (2 ml), the solution is stirred at room 
temperature for 4 hours. The reaction mixture is poured in water (100 ml) 
and rinsed with ethyl acetate (40 ml). The aqueous layer is adjusted to pH 
2.0 with diluted phosphoric acid and extracted with a saturated aqueous 
solution of sodium chloride. The solution is dried over magnesium sulfate 
and concentrated. Then, upon addition of ether, 
7.beta.-(2-thienylacetamido)-3-[2-(N-carboethoxysulfamoyl)benzoyloxy]methy 
l-3-cephem-4-carboxylic acid crystallizes with one mole of ether. 
Yield 3.8 g. 
IR(KBr): cm.sup.-1 1790, 1748, 1698, 1652 
NMR(d.sub.6 -DMSO): .delta. 1.0-1.2(9H, m, --COOCH.sub.2 CH.sub.3 & 
(CH.sub.3 CH.sub.2).sub.2 O), 3.35(4H, q, (CH.sub.3 CH.sub.2).sub.2 O), 
3.63(2H, broad, 2--CH.sub.2),3.75 (2H, s, --CH.sub.2 CONH--), 4.01(2H, q, 
--COOCH.sub.2 CH.sub.3), 5.07(1H, d, J=5 Hz, 6-H), 5.13(2H, ABq, J=13 Hz, 
3--CH.sub.2), 5.70(b 1H, dd, J=5 & 8 Hz, 7-H), 
##STR11## 
9.07(1H, d, J=8 Hz, --CONH--) 
EXAMPLE 5 
The reaction of Example 4 is repeated using 
7.beta.-phenylacetamido-3-hydroxymethyl-3-cephem-4-carboxylic acid sodium 
salt (3.7 g) in place of 
7.beta.-(2-thienylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic acid 
sodium salt. This procedure provides crystals of 
7.beta.-phenylacetamido-3-[2-(N-carboethoxysulfamoyl)benzoyloxy]methyl-3-c 
ephem-4-carboxylic acid-ether(one mole). Yield 4.0 g. 
IR(KBr): cm.sup.-1 1794, 1750, 1701, 1650 
NMR(d.sub.6 -DMSO): .delta. 1.0-1.2(9H, m, --COOCH.sub.2 CH.sub.3 & 
(CH.sub.3 CH.sub.2).sub.2 O), 3.35(4H, q, (CH.sub.3 CH.sub.2).sub.2 O), 
3.52(2H, --CH.sub.2 CO--), 3.62(2H, broad, 2--CH.sub.2), 4.01(2H, q, 
--COOCH.sub.2 CH.sub.3), 5.05(1H, d, J=5 Hz, 6-H), 5.17(2H, ABq, J=13 Hz, 
3--CH.sub.2), 5.69(1H, dd, J=5 & 8 Hz, 7-H), 
##STR12## 
9.05(1H, d, J=8 Hz --CONH) 
Analysis: C.sub.30 H.sub.35 N.sub.3 O.sub.11 S.sub.2 Calcd.(%): C, 53.17; 
H5.21; N, 6.20; S, 9.46; Found (%): C, 53.20; H, 5.34; N, 6.23; S, 9.65. 
EXAMPLE 6 
In water (50 ml) is suspended 
7.beta.-phenylacetamido-3-[2-(N-carboethoxysulfamoyl)benzoyloxy]methyl-3-c 
ephem-4-carboxylic acid (3.0 g) together with 
5-mercapto-1-methyl-1H-tetrazole (1.0 g). The mixture is adjusted to pH 
5.5 with sodium hydrogen carbonate and the resultant solution is heated at 
60.degree. C. for 50 minutes. The solution is brought to pH 2.0 with 
dilute phosphoric acid and extracted with ethyl acetate (100 ml). The 
extract is rinsed with a saturated aqueous solution of sodium chloride, 
dried and concentrated. Then, following the addition of ether, the 
precipitate is recovered by filtration. By the above procedure is obtained 
7.beta.-phenylacetamido-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4 
-carboxylic acid (1.8 g). 
IR(KBr): cm.sup.-1 3270, 1785, 1733, 1662, 1628, 1542 
NMR(d.sub.6 -DMSO): .delta. 3.55(2H, s, 
##STR13## 
3.60(2H, broad, 2--CH.sub.2), 3.92(3H, s, --N--CH.sub.3), 4.26(2H, broad, 
3--CH.sub.2 --), 5.00(1H, d, J=5 Hz, 6-H), 5.60(1H, q, J=5 & 8 Hz, 7-H), 
7.23(5H, s), 8.98(1H, d, J=8 Hz, --CONH) 
EXAMPLE 7 
In N,N-dimethylformamide (40 ml) is dissolved 
7.beta.-(2-thienylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic acid 
sodium salt (3.8 g), followed by the addition of N-tosylphthalimide (4.0 
g) and triethylamine (2.0 ml) in that order. The mixture is stirred at 
room temperature for 4 hours. The reaction mixture is poured in water (100 
ml) and rinsed with ethyl acetate. The aqueous layer is adjusted to pH 2.0 
with dilute phosphoric acid and extracted with ethyl acetate. The extract 
is dried and, then, a solution of sodium 2-ethylhexanoate in isopropyl 
alcohol is added. By this procedure is obtained 
7.beta.-(2-thienylacetamido)-3-[2-(N-tosylcarbamoyl)benzoyloxy]methyl-3-ce 
phem-4-carboxylic acid sodium salt (4.0 g). 
IR(KBr): cm.sup.-1 1769, 1695, 1620 
NMR(d.sub.6 -DMSO): .delta. 2.27(3H, s, --CH.sub.3), 3.48(2H, broad, 
2--CH.sub.2), 3.74(2H, s, CH.sub.2 CONH), 4.97(1H, d, 6-H), 5.00(2H, ABq, 
J=13 Hz, 3--CH.sub.2), 5.58(1H, q, J=5 & 8 Hz, 7-H), 6.8-7.9(11H, m), 
9.02(1H, d, J=8 Hz, 7--CONH) 
EXAMPLE 8 
In N,N-dimethylformamide (10 ml) is dissolved 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-hydroxymethyl-3-cephem-4-c 
arboxylic acid sodium salt (1.3 g), followed by the addition of 
N-carboethoxysaccharin (0.8 g) and triethylamine (0.4 ml). The mixture is 
stirred at room temperature for 4 hours. It is then poured in water (50 
ml) and rinsed with ethyl acetate. The aqueous layer is adjusted to pH 2.0 
with dilute phosphoric acid and extracted with ethyl acetate. The extract 
is dried and concentrated, followed by addition of ether. By this 
procedure is obtained 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-[2-(N-carboethoxysulfamoyl 
)benzoyloxy]methyl-3-cephem-4-carboxylic acid (1.4 g) as a powder. 
NMR(d.sub.6 -DMSO): .delta. 1.10(3H, t, CH.sub.2 CH.sub.3), 1.3-2.4(6H, m, 
--(CH.sub.2).sub.3 --), 3.60(2H, broad, 2--CH.sub.2), 4.01(2H, q, CH.sub.2 
CH.sub.3), 4.73(1H, t, J=7 Hz, 
##STR14## 
5.03(1H, d, J=5 Hz, 6-H), 5.15(2H, Abq, J=13 Hz, 3--CH.sub.2), 5.65(1H, q, 
J=5 & 8 Hz, 7-H), 7.48-8.1(8H, m), 8.75(1H, d, J=8 Hz, --CONH--) 
EXAMPLE 9 
In water (8 ml) is added 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-[2-(N-carboethoxysulfamoyl 
)benzoyloxy]methyl-3-cephem-4-carboxylic acid (0.8 g), which is then 
dissolved by the addition of sodium hydrogen carbonate. Then, 
5-mercapto-1-methyl-1H-tetrazole (0.15 g) is added, followed by addition 
of a further amount of sodium hydrogen carbonate to adjust to pH 5.3. The 
mixture is heated on a water bath at 60.degree. C. for 50 minutes, after 
which time it is brought to pH 2.0 with dilute phosphoric acid and 
extracted with ethyl acetate (20 ml). The extract is dried and 
concentrated, followed by the addition of ether to the residue. By the 
above procedure is obtained 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-(1-methyl-1H-tetrazol-5-yl 
)thiomethyl-3-cephem-4-carboxylic acid (0.6 g). In NMR spectrum, this 
product is in good agreement with the product obtained in Example 2. 
EXAMPLE 10 
In a mixture of N,N-dimethylformamide (16 ml) and water (4 ml) is dissolved 
7.beta.-(D-5-benzamido-5-carboxyvaleramido)-3-hydroxymethyl-3-cephem-4-car 
boxylic acid disodium salt (2.6 g), followed by the addition of 
N-tosylphthalimide (2.3 g) and triethylamine (0.7 g). The mixture is 
stirred at room temperature for 2 hours. The reaction mixture is poured in 
water, rinsed with ethyl acetate, adjusted to pH 2.0 with dilute 
phosphoric acid and extracted with ethyl acetate. After drying, the 
solvent is distilled off and the resultant residue is made into a powder 
by a rinse with ether. By the above procedure is obtained 
7.beta.-(D-5-benzamido-5-carboxyvaleramido)-3-[2-(N-tosylcarbamoyl)benzoyl 
oxy]methyl-3-cephem-4-carboxylic acid (3.5 g). 
IR(KBr): cm.sup.-1 1780, 1725 
NMR(d.sub.6 -DMSO): .delta. 1.5-2.5(6H, m, --(CH.sub.2).sub.3 --), 2.34(3H, 
s, --CH.sub.3), 3.48(2H, ABq, J=18 Hz, 2--CH.sub.2), 4.36(1H, broad, 
##STR15## 
4.93(2H, ABq, J=13 Hz, 3--CH.sub.2), 5.01(1H, d, J=5 Hz, 6-H), 
5.64(1H, q, J=5 & 8 Hz, 7-H), 7.2-8.0(13H, m, arom-H), 8.51(1H, d, J=8 Hz, 
--CONH--), 8.79(1H, d, J=8 Hz, --CONH--) 
EXAMPLE 11 
With the addition of sodium hydrogen carbonate, 
7.beta.-(D-5-benzamido-5-carboxyvaleramido)-3-[2-(N-tosylcarbamoyl)benzoyl 
oxy]methyl-3-cephem-4-carboxylic acid (1.6 g) is dissolved in water (20 
ml), followed by the addition of a further amount of sodium hydrogen 
carbonate to adjust the solution to pH 5.2. The solution is heated at 
60.degree. C. for 50 minutes and, after cooling, the mixture is diluted 
with a saturated aqueous solution of sodium chloride (40 ml) and adjusted 
to pH 1.5 with dilute hydrochloric acid. The resultant solid precipitate 
is recovered by filtration, rinsed with a saturated aqueous solution of 
sodium chloride and dissolved in a mixture of ethyl acetatetetrahydrofuran 
(2:1)(40 ml) and water (10 ml). After separation, the organic layer is 
dried and the solvent is distilled off. The residue is rinsed with ether 
to obtain a powder. By the above procedure is obtained 
7.beta.-(D-5-benzamido-5-carboxyvaleramido)-3-(1-methyl-1H-tetrazol-5-yl)t 
hiomethyl-3-cephem-4-carboxylic acid (1.1 g). 
IR(KBr): cm.sup.-1 3340, 1783, 1730, 1645, 1535 
NMR(d.sub.6 -DMSO): .delta. 1.50-2.0(4H, m), 2.05-2.45(2H, m), 3.70(2H, 
broad), 3.93(3H, s), 4.15-4.55(3H, m), 5.10(1H, d, J=5 Hz), 5.66(1H, q, 
J=5 & 9 Hz), 7.32-7.97(5H, m), 8.43(1H, d, J=8 Hz), 8.73(1H, d, J=9 Hz) 
EXAMPLE 12 
In N,N-dimethylformamide (30 ml) is dissolved 
7.beta.-(2-thienylacetamido)-3-hydroxymethyl-3-cephem-4-carboxylic acid 
sodium salt (3.8 g), followed by addition of N-carboethoxysuccinimide (2.5 
g) and triethylamine (1.5 ml). The mixture is stirred at room temperature 
for 20 hours. The reaction mixture is poured in ice-water and rinsed with 
ethyl acetate (50 ml). The aqueous layer is adjusted to pH 2.0 with 
diluted phosphoric acid, extracted with ethyl acetate and dried. Then, a 
solution of sodium 2-ethylhexanoate in isopropyl alcohol is added. The 
precipitate is 
7.beta.-(2-thienylacetamido)-3-[3-(N-carboethoxycarbamoyl)propionyloxy]met 
hyl-3-cephem-4-carboxylic acid sodium salt (32 g). 
NMR(D.sub.2 O): .delta. 1.33(3H, t, CH.sub.3 --), 2.80(4H, broad, 
--(CH.sub.2).sub.2 --), 3.44 (2H, ABq, J=18 Hz, 2--CH.sub.2), 3.81(2H, s, 
--CH.sub.2 CO--), 4.25(2H, q, --CH.sub.2 --), 5.07(1H, d, J=5 Hz, 6-H), 
5.70(1H, d, J=5Hz, 7-H), 
##STR16## 
EXAMPLE 13 
In acetonitrile (10 ml) is dissolved 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-hydroxymethyl-3-cephem-4-c 
arboxylic acid ditriethylamine salt (705 mg), followed by the addition of 
triethylamine (202 mg), 5-mercapto-1-methyl-1H-tetrazole (232 mg) and 
N-carboethoxyisatin (438 mg) in the order mentioned. The reaction is 
carried out at room temperature for 3 hours, after which time the solvent 
is distilled off under reduced pressure. The residue is dissolved in 
water-ethyl acetate, adjusted to pH 2.0 and extracted with ethyl acetate. 
The ethyl acetate solution is rinsed with water and, following the 
addition of water, is adjusted to pH 5.0 with an aqueous solution of 
sodium hydrogen carbonate. After separation, the aqueous solution is 
concentrated and the residue is subjected to column chromatography on 
Amberlite XAD-2. Elution is carried out with a mixture of water-methanol 
and the eluate is freeze-dried. By the above procedure is obtained 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-(1-methyl-1H-tetrazol-5-yl 
)thiomethyl-3-cephem-4-carboxylic acid monosodium salt (512 mg). 
IR(KBr): cm.sup.-1 1768, 1710, 1665, 1610, 1535 
NMR(D.sub.2 O): .delta. 1.30-2.60(6H, m), 3.34(2H, ABq, J=20 Hz, 
2--CH.sub.2), 3.99(3H, s, N--CH.sub.3), 4.10(2H, ABq, J=14 Hz, 
3--CH.sub.2), 4.50(1H, broad), 4.97(1H, d, J=5 Hz, 6-H), 5.53(1H, d, 
J=5Hz, 7-H), 7.86(4H, s, arom-H) 
EXAMPLE 14 
In DMF (10 ml) is dissolved 
7.beta.-phenylacetamido-3-hydroxymethyl-3-cephem-4-carboxylic acid 
triethylamine salt (900 mg), followed by the addition of triethylamine 
(0.4 ml) and N-carboethoxyisatin (876 mg). The mixture is stirred at room 
temperature for 2 hours, after which time the DMF is distilled off under 
reduced pressure. 
Following the addition of water and ethyl acetate, the residue is adjusted 
to pH 2.0. The aqueous layer is taken, adjusted to pH 4.0 with an aqueous 
solution of sodium hydrogen carbonate and subjected to column 
chromatography on Dowex 1.times.2 (AcO.sup.-) and the resultant aqueous 
solution is freeze-dried. By this procedure is obtained 
N-(7.beta.-phenylacetamido-3-cephem-3-ylmethyl)triethylammonium-4-carboxyl 
ate (620 mg). 
IR(KBr): cm.sup.-1 1775, 1665, 1615, 1545, etc. 
NMR(D.sub.2 O): .delta. 1.29(9H, t, J=6 Hz), 2.65-4.25(12H, m), 4.55(1H, 
broad), 5.08(1H, d, J=5 Hz, 6-H), 5.62(1H, d, J=5 Hz, 7-H), 7.25(5H, s, 
arom-H) 
EXAMPLE 15 
In dimethylformamide (4.0 ml) is suspended deacetylcephalosporin C sodium 
monohydrate (purity 90%, 826 mg), and under cooling with ice, concentrated 
hydrochloric acid (0.33 ml) is added. To the resultant clear solution is 
added DMF (8.0 ml) together with triethylamine (2.1 ml) and 
5-mercapto-1-methyl-1H-tetrazole (464 mg). Then, at room temperature, 
N-carboethoxyisatin (1.752 g) is added in small installments. Thereafter, 
the reaction is conducted at room temperature for 4 hours. After the 
reaction is completed, the DMF is distilled off under reduced pressure. 
Following the addition of water and ethyl acetate, the residue is adjusted 
to pH 2.0. The ethyl acetate layer is separated and diluted with water and 
adjusted to pH 7.0 with an aqueous solution of sodium hydrogen carbonate. 
After separation, the aqueous solution is concentrated and subjected to 
column chromatography on Amberlite XAD-2. Elution is carried out with a 
solvent mixture of water and methanol and the eluate is freeze-dried. By 
the above procedure is obtained 
7.beta.-[D-5-(2-ethoxycarbonylamino)-phenylglyoxamido-5-carboxyvaleramido] 
- 3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid 
disodium salt (1.051 g). 
IR(KBr): cm.sup.-1 1765, 1745, 1665, 1645, 1607, 1590, 1530 
NMR(D.sub.2 O): .delta. 1.20(3H, t, J=8 Hz), 1.40-2.15(4H, m), 2.20-2.60 
(2H, m), 3.43(2H, ABq, J=18 Hz, 2--CH.sub.2), 3.88(3H, s, N--CH.sub.3), 
3.75-4.50(5H, m), 5.04(1H, d, J=5 Hz, 6-H), 5.56(1H, d, J=5 Hz, 7-H), 
7.05-8.05(4H, m, arom-H) 
EXAMPLE 16 
(1) A mixture of dichloromethane (300 ml), triethylamine (27 ml) and 
dimethylaniline (100 ml) is previously cooled to 10.degree. C. and, then, 
7-[D-5-(phthalimido-5-carboxy 
valeramido]-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic 
acid (50 g) is added and dissolved. Then, upon addition of 
dichlorodimethylsilane (36 ml), the internal temperature increases to 
27.degree. C. The mixture is stirred at this temperature for 30 minutes, 
after which time it is cooled to an internal temperature of -35.degree. C. 
Then, phosphorus pentachloride (32.4 g) is added. The mixture is stirred 
at -25.degree. C. for 40 minutes and, then, at -35.degree. C., 
thioacetamide (20 g) is added. The mixture is stirred at 
-20.degree.--25.degree. C. for 40 minutes and, then, at -30.degree. C., 
methanol (200 ml) is gently added dropwise. Then, at the same temperature, 
sulfur monochloride (17 ml) is gently added dropwise. The mixture is 
stirred for 20 minutes, after which time water (200 ml) was added. It is 
then adjusted to pH 3.2 with 40% aqueous potassium carbonate solution and 
stirred for 60 minutes. The resultant crystals are recovered by filtration 
and rinsed with water and acetone. The crude crystals thus obtained are 
suspended in 10% hydrochloric acid (230 ml) and stirred at 30.degree. C. 
for 1 hour. The insolubles are filtered off and, after cooling to 
5.degree.-10.degree. C., the filtrate is adjusted to pH 3.3 with potassium 
carbonate, followed by stirring for 1 hour. The resultant crystals are 
recovered by filtration, rinsed with water and acetone, and dried over 
phosphorus pentoxide. By the above procedure is obtained 
7-amino-3-(1-methyltetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid 
(17.0 g) 
IR(KBr): 1795 cm.sup.-1 
NMR(in D.sub.2 O+NaHCO.sub.3): .delta. 3.61 & 3.98(Abq, J=18 Hz, 
2--CH.sub.2), 4.21(s, tetrazole --CH.sub.3), 5.21(d, J=4.5 Hz, 6-H), 5.60 
(d, J=4.5 Hz, 7-H) 
(2) Chlorine gas (2.8 g) is bubbled through a solution of diketene (3.3 g) 
in methylene chloride (160 ml) under cooling and stirring to maintain the 
internal temperature at -25.degree.--35.degree. C. over a period of 100 
minutes, after which time the mixture is further stirred at that 
temperature for 30 minutes. Separately, 
7-amino-3-(1-methyltetrazol-5-yl)-thiomethyl-3-cephem-4-carboxylic acid 
(10.0 g) and dibutylamine (7.9 g) are dissolved in methylene chloride (60 
ml) and the solution is cooled to -10.degree. C. 
To this is added dropwise the above reaction mixture over a period of 30 
minutes, with cooling and stirring to maintain the temperature of the 
solution at -10.degree. C. to -20.degree. C. Then, the mixture is further 
stirred at that temperature for 40 minutes. 
Thin-layer chromatography of this reaction mixture demonstrates the 
presence of 
7.beta.-(4-chloro-3-oxobutyrylamido)-3-(1-methyltetrazol-5-yl)thiomethyl-3 
-cephem-4-carboxylic acid. In this reaction mixture is dissolved thiourea 
(4.64 g) and the internal temperature is gradually increased to 17.degree. 
to 19.degree. C. Then, the mixture is stirred at that temperature, 
whereupon crystals separates. These crystals are collected by 
suction-filtration, rinsed with methylene chloride (30 ml) and dried. By 
the above procedure is obtained 
2-(2-imino-4-thiazolin-4-yl)acetamido-3-(1-methyltetrazol-5-yl)thiomethyl- 
3-cephem-4-carboxylic acid. (Melting point: 176.degree.-180.degree. C. 
(decomp.) 
IR(KBr): cm.sup.-1 1762, 1662 
NMR(d.sub.6 -DMSO) .delta. 3.39(s, CH.sub.2 CO) 3.55 & 3.77(ABq, J=18 Hz, 
2--CH.sub.2), 3.90(s, tetrazole 1--CH.sub.3), 4.21 & 4.36(ABq, J=14 Hz,) 
3--CH.sub.2), 5.03(d, J=5 Hz, 6-H), 5.66(dd, J=9 & 5 Hz, 7-H), 6.23(s, 
thiazoline 5-H), 6.2-7.1(m, NHX2), 8.85(d, J=9 Hz, CONH) 
EXAMPLE 17 
In acetonitrile (14 ml) is dissolved 
7.beta.-phenylacetamido-3-hydroxymethyl-3-cephem-4-carboxylic acid 
triethylamine salt (900 mg), followed by the addition of triethylamine 
(200 mg), 5-mercapto-1-methyl-1H-tetrazole (464 mg) and 
N-(methylcarbamoyl)isatin (916 mg). The mixture is stirred at 30.degree. 
C. overnight. The solvent is then distilled off and the residue is 
dissolved by the addition of water and ethyl acetate. After separation, 
the water layer is taken, rinsed once with ethyl acetate and rendered 
acidic with hydrochloric acid. Following the addition of ethyl acetate, 
the mixture is stirred for 30 minutes. After separation, the organic layer 
is taken and rinsed with an aqueous solution of sodium chloride. It is 
extracted with an aqueous solution of sodium hydrogen carbonate. The water 
layer is purified by column chromatography on Sephadex LH-20. The 
fractions rich in the contemplated compound are pooled, concentrated, 
rendered acidic with phosphoric acid and extracted with ethyl acetate. 
After drying, the extract is concentrated, followed by addition of ether 
to the residue. The procedure provides 
7.beta.-phenylacetamido-3-(1-methyl-1H-tetrazol-5-yl)thiomethyl-3-cephem-4 
-carboxylic acid. The NMR spectrum of this product is in good agreement 
with that of the compound obtained in Example 6. 
EXAMPLE 18 
In dichloromethane (100 ml) is dissolved triethylamine 
7.beta.-phenylacetamido-3-hydroxymethyl-3-cephem-4-carboxylate (4.5 g), 
followed by the addition of triethylamine (1.4 ml) and 
N-carboethoxysaccharin (3.0 g). The mixture is stirred at room temperature 
overnight. The solvent is distilled off under reduced pressure and the 
residue is stirred with water and ethyl acetate. After separation, the 
water layer is taken, rendered acidic with phosphoric acid and extracted 
with ethyl acetate. After drying, the solvent is distilled off, followed 
by addition of ether to the residue. The above procedure provides crystals 
(6.0 g). This product is in good agreement with the product obtained in 
Example 4 in infrared spectrum. 
EXAMPLE 19 
In N,N-dimethylformamide (40 ml) is dissolved disodium 
7.beta.-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-hydroxymethyl-3-c 
ephem-4-carboxylate (5.2 g) and, under cooling with ice, 
N-carboethoxysaccharin (3.2 g) and triethylamine (1.6 ml) are added. The 
mixture is stirred at room temperature for 2 hours. The reaction mixture 
is poured in icewater and rinsed twice with ethyl acetate. The water layer 
is adjusted to pH 2.0 and extracted with ethyl acetate. The organic layer 
is rinsed twice with water, dried over sodium sulfate and concentrated. 
Following the addition of ether, the solution is cooled with ice and the 
resultant precipitate is recovered by filtration. By the above procedure 
is obtained 
7.beta.-[D-5-(p-t-butylbenzamido)-5-carboxyvaleramido]-3-[2-(N-carboethoxy 
sulfamoyl)benzoyloxy]-methyl-3-cepham-4-carboxylic acid (5.3 g). 
NMR(d.sub.6 -DMSO): .delta. 1.09(3H, t, --CH.sub.2 CH.sub.3), 1.5-2.5(6H, 
m), 3.61(2H, broad, 2--CH.sub.2), 3.99(2H, q, --CH.sub.2 CH.sub.3), 
4.35(1H, broad, --CH--), 4.99 & 5.32 (2H, ABq, J=13 Hz, 3--CH.sub.2), 
5.05(1H, d, J=5 Hz, 6-H), 5.66(1H, dd, J=5 & 8,7-H), 7.3-8.1(8H, m), 
8.42(1H, d, --CONH--), 8.78(1H, d, --CONH--) 
EXAMPLE 20 
In dichloromethane (120 ml) is suspended 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-[2-(N-carboethoxycarbamoyl 
)-benzoyloxy]methyl-3-cephem-4-carboxylic acid (14.5 g) and, at a 
temperature not exceeding 10.degree. C., the suspension is dissolved by 
the addition of triethylamine (12 ml). Following the addition of 
N,N-dimethylaniline (20 ml) and dimethyldichlorosilane (8.9 ml), the 
mixture is stirred at 20.degree.-25.degree. C. for 30 minutes. After 
cooling to -30.degree. C., phosphorus pentachloride (9.85 g) is added. The 
reaction is conducted at -25.degree. C. for 30 minutes and, then, at a 
temperature not exceeding -20.degree. C., methanol (50 ml) is added 
dropwise. Then, at -15.degree.--10.degree. C., the reaction is further 
carried out for 20 minutes. Following the addition of ice-water (100 ml), 
the mixture is stirred for 5 minutes. The reaction mixture is separated 
and the water layer is taken, rinsed with dichloromethane and brought to 
pH 3.5 with 40% aqueous potassium carbonate solution. The precipitate is 
recovered by filtration, rinsed with water, 50% water-methanol and 
acetone. By the above procedure is obtained 
7.beta.-amino-3-[2-(N-carboethoxycarbamoyl)benzoyloxy]-methyl-3-cephem-4-c 
arboxylic acid. 
IR(KBr): cm.sup.-1 1780, 1723 
NMR(D.sub.2 O+NaOD): .delta. 1.30(3H, t, CH.sub.2 CH.sub.3), 3.54(2H, ABq, 
J=18 Hz, 2--CH.sub.2), 4.25(2H, q, --CH.sub.2 CH.sub.3), 7.4-8.2(4H, m, 
arom-H) 
EXAMPLE 21 
(1) In dimethylformamide (20 ml) is dissolved sodium 
7.beta.-(t-butoxycarbonyl)amino-3-hydroxymethyl-3-cephem-4-carboxylate 
(3.20 g), followed by the addition of N-carboethoxysaccharin (3.20 g) and 
triethylamine (2.0 ml). The solution is stirred for one hour at room 
temperature and, after the addition of triethylamine (2.0 ml), stirred 
further for one hour. The solution is poured into toluene (1 L) and the 
resultant precipitate is collected by filtration, washed with toluene and 
dried. The powder thus obtained is dissolved in water (30 ml). After the 
addition of ethyl acetate (200 ml), the solution is adjusted to pH 4.0 
with diluted phosphoric acid. After separation, the ethyl acetate solution 
is washed with water, dried over anhydrous sodium sulfate and 
concentrated. To the concentrate (30 ml), ether (200 ml) is added under 
stirring and the resultant precipitate is filtered off. The filtrate is 
concentrated under reduced pressure and to the concentrate (30 ml) is 
added carbon tetrachloride (100 ml). The resultant precipitate is 
collected by filtration, washed with carbon tetrachloride and dried over 
phosphorus pentoxide. The procedure provides 
7.beta.-(t-butoxycarbonyl)amino-3-[2-(N-carboethoxysulfamoyl)benzoyloxy]me 
thyl-3-cephem-4-carboxylic acid (2.50 g). 
IR(KBr): cm.sup.-1 3400-3250, 1790 1733 
NMR(d.sub.6 -DMSO): .delta. 1.12(3H, t, J=7 Hz, --CH.sub.2 CH.sub.3), 
1.42(9H, s, --C(CH.sub.3).sub.3), 3.62(2H, broad, 2--CH.sub.2), 4.03(2H, 
q, J=7 Hz, --CH.sub.2 CH.sub.3), 5.05 (1H, d, J=5 Hz, 6-H), 5.18(2H, ABq, 
J=13 Hz, 3--CH.sub.2), 5.48(1H, q, J=5 & 8 Hz, 7-H), 7.57-8.11(4H, m, 
arom-H) 
(2) 
7.beta.-(t-Butoxycarbonyl)amino-3-[2-(N-carboethoxysulfamoyl)benzoyloxy]me 
thyl-3-cephem-4-carboxylic acid (2.2 g) is dissolved in ice cooled solution 
of trifluoroacetic acid (20 ml) and the mixture is stirred for 20 minutes. 
Then, trifluoroacetic acid is taken off under reduced pressure and viscous 
residue is obtained. The residue becomes solid upon addition of ether (50 
ml). The solid is triturated, collected by filtration, washed with ether 
and dried over phosphorus pentoxide. The procedure provides 
7.beta.-amino-3-[2-(N-carboethoxysulfamoyl)benzoyloxy]methyl-3-cephem-4-ca 
rboxylic acid (1.55 g). 
IR(KBr): cm.sup.-1 3230, 1771, 1730 
NMR(d.sub.6 -DMSO): .delta. 1.11(3H, t, J=7 Hz, --CH.sub.2 CH.sub.3), 
3.66(2H, broad, 2--CH.sub.2), 4.00(2H, q, J=7 Hz, --CH.sub.2 CH.sub.3), 
4.94(2H, ABq, J=14 Hz, 3--CH.sub.2), 5.02(1H, d, J=5 Hz, 6--H), 5.26(1H, 
d, J=5 Hz, 7--H), 7.55-8.13(4H, m, arom-H) 
EXAMPLE 22 
in water (2 ml) containing sodium salt of 5-mercapto-1H-1,2,3-triazole (120 
mg) and sodium hydroxide (40 mg) is dissolved under ice cooling 
7.beta.-amino-3-[2-(N-carboethoxycarbamoyl)benzoyloxy]methyl-3-cephem-4-ca 
rboxylic acid (450 mg) and to the mixture is added diluted hydrochloric 
acid to adjust its pH to 5.5 under stirring, followed by further stirring 
for one hour at 60.degree. C. To the solution, is added methanol (5 ml) 
and the mixture is allowed to cool to the room temperature. The cooled 
mixture is adjusted its pH to 3.9 by adding diluted hydrochloric acid 
under stirring and the resultant mixture is further stirred for one hour 
under ice-cooling. The precipitate is collected by filtration and washed 
with water and methanol in this order and then dried over phosphorus 
pentoxide. The procedure provides 
7.beta.-amino-3-(1H-1,2,3-triazol-5-yl)thiomethyl-3-cephem-4-carboxylic 
acid (193 mg). 
IR(KBr): cm.sup.-1 1800, 1525 
EXAMPLE 23 
While a solution of diketene (0.01 ml) in dichloromethane (1 ml) is stirred 
at -50.degree.--40.degree. C., 1.0 M bromine solution in dichloromethane 
(1.40 ml) is added dropwise for five minutes and stirred for 20 minutes. 
Separately, 
7.beta.-amino-3-[2-(N-carboethoxysulfamoyl)benzoyloxy]methyl-3-cephem-4-ca 
rboxylic acid (475 mg) is suspended in dichloromethane (3 ml) and cooled to 
-40.degree. C., followed by the addition of triethylamine (0.42 ml) in 
dichloromethane (3 ml). This solution is added to the above reaction 
mixture at -40.degree.--30.degree. C. After stirring for 10 minutes, the 
mixture is further stirred under cooling with ice for 30 minutes. Then, 
dichloromethane is distilled off under reduced pressure. To the residue, 
10% phosphoric acid (5 ml), water (10 ml), tetrahydrofuran (2 ml) and 
ethyl acetate (10 ml) are added and the mixture is stirred vigorously. The 
organic layer is washed with a saturated solution of sodium chloride and 
dried. The solvent is distilled off and ether (20 ml) is added. The 
procedure provides 
7.beta.-(4-bromo-3-oxobutylamido)-3-[2-(N-carboethoxysulfamoyl)benzoyloxy] 
methyl-3-cephem-4-carboxylic acid (0.5 g). 
NMR(d.sub.6 -DMSO): .delta. 1.1(3H, t, --CH.sub.2 CH.sub.3), 3.63(4H, 
broad, --CH.sub.2 CO, 2--CH.sub.2), 4.02(2H, q, --CH.sub.2 CH.sub.3), 
4.39(2H, s, BrCH.sub.2 --), 5.05 (1H, d, J=5 Hz, 6--H), 5.17(2H, ABq, J=13 
Hz, 3--CH.sub.2), 5.70 (1H, dd, J=5 & 8Hz, 7--H), 7.6-8.1(4H, m, 
##STR17## 
9.06(1H, d, J=8Hz, --CONH--) 
EXAMPLE 24 
7.beta.-(Thienylacetamido)-3-[2-(N-carboethoxysulfamoyl)benzoyloxy]methyl-3 
-cephem-4-carboxylic acid-etherate (619 mg), sodium azide (100 mg) and 
sodium bicarbonate (84 mg) are dissolved in phosphate buffer solution (pH 
6.4) (10 ml) and the mixture is stirred at 60.degree. C. for one hour. 
After cooling, the reaction mixture is acidified by phosphoric acid and 
extracted with ethyl acetate. The ethyl acetate layer is dried and 
concentrated under reduced pressure. The residue is triturated with ether. 
The procedure provides 
7.beta.-(thienylacetamido)-3-azidomethyl-3-cephem-4-carboxylic acid. 
IR(KBr): cm.sup.-1 2110 
EXAMPLE 25 
7.beta.-Phenylacetamido-3-hydroxymethyl-3-cephem-4-carboxylic acid 
triethylamine salt (450 mg) is dissolved in dichloromethane (5 ml), 
followed by the addition of N-carboethoxy-4-nitrophthalimide (330 mg) and 
triethylamine (0.14 ml). The mixture is stirred at room temperature 
overnight. The solvent is distilled off and to the residue, ethyl acetate 
(20 ml) and water (20 ml) are added. After separation, aqueous layer is 
washed with ethyl acetate and acidified to pH 2.0 and extracted with ethyl 
acetate. The ethyl acetate layer is dried and concentrated. Ether is added 
to the residue and the resultant powder is collected by filtration. The 
procedure provides 7.beta.-phenylacetamido-3-[2-(N-ethoxycarbamoyl)-4(or 
5)-nitrobenzoyloxy]methyl-3-cephem-4-carboxylic acid (420 mg). 
IR(KBr): cm.sup.-1 1776, 1735 
NMR(d.sub.6 -DMSO): .delta. 1.17(3H, t, CH.sub.2 CH.sub.3), 3.57(2H, s, 
--CH.sub.2 CO--), 3.65(2H, board, 2--CH.sub.2), 4.06(2H, q, --CH.sub.2 
CH.sub.3), 5.01 & 5.35(2H, ABq, J=13 Hz, 3--CH.sub.2), 5.13(1H, d, J=5 Hz, 
6--H), 5.73(1H, dd, J=5 & 8Hz, 7--H) 
##STR18## 
9.09(1H, d, --CONH--) 
EXAMPLE 26 
In acetonitrile (10 ml) is dissolved 
7.beta.-{D-5-(p-toluenesulfonamido)-5-carboxyvaleramido}-3-hydroxymethyl-3 
-cephem-4-carboxylic acid ditriethylamine salt (729 mg), followed by the 
addition of triethylamine (202 mg), 5-mercapto-1-methyl-1H-tetrazole (232 
mg) and N-carboethoxyisatin (438 mg) in the order mentioned. The solution 
is reacted at room temperature for 3 hours and then, the solvent is 
distilled off under reduced pressure. The residue is dissolved in 
water-ethyl acetate, adjusted to pH 2.0 and extracted with ethyl acetate. 
The organic layer is washed with water and, following the addition of 
water, is adjusted to pH 5.0 with sodium hydrogen carbonate. Aqueous 
solution is concentrated and the residue is subjected to column 
chromatography on Amberlite XAD-2. Elution is carried out with a mixture 
of water-methanol and the eluate is collected and concentrated. The 
concentrate is acidified and extracted with ethyl acetate. The extract is 
dried and concentrated. 
The procedure provides 
7.beta.-{D-5-(p-toluenesulfonamido)-5-carboxyvaleramido}-3-(1-methyl-1H-te 
trazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid. 
IR(KBr): cm.sup.-1 3275, 1780, 1727, 1635, 1535 
NMR(d.sub.6 -DMSO): .delta. 1.45-1.78(4H, m), 2.0-2.3(2H, m), 2.41(3H, s, 
--CH.sub.3), 3.71(2H, broad, 2--CH.sub.2), 3.95(3H, s, --NCH.sub.3), 
4.28(2H, broad), 5.06(1H, d, J=5 Hz, 6--H), 5.62(1H, dd, J=5.0 & 9.0 Hz, 
7--H) 
##STR19## 
7.91(1H, d, J=9.0 Hz), 8.68(1H, d, J=9.0Hz) 
EXAMPLE 27 
In a mixture of water (50 ml) and tetrahydrofuran (30 ml) is dissolved 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-[2-(N-carboethoxycarbamoyl 
)benzoyloxy]methyl-3-cephem-4-carboxylic acid (7.23 g) together with 
2-carbamoylmethylthio-5-mercapto-1,3,4-thiadiazole (2.28 g) and sodium 
bicarbonate (2.20 g). The solution is adjusted to pH 5.8 and reacted at 
60.degree. C. for one hour. After cooling, the solution is adjusted to pH 
5.0, washed twice with ethyl acetate and brought down to pH 2.0 with 
diluted hydrochloric acid. Then, it is extracted three times with a 
solvent mixture of ethyl acetate and tetrahydrofuran (2:1) and the organic 
layer is washed with a saturated aqueous solution of sodium chloride and 
dried over magnesium sulfate. The solvent is distilled off and the residue 
is treated with ethyl acetate. The resultant powder is collected by 
filtration, washed with ethyl acetate and dried. The procedure provides 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-(2-carbamoylmethylthio-1,3 
,4-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid (5.70 g). 
IR(KBr): cm.sup.-1 3430, 3340, 1776, 1717, 1680, 1535 
NMR(d.sub.6 -DMSO): .delta. 1.30-2.40(6H, m), 3.57(2H, br), 4.40(2H, s), 
4.32(2H, ABq, J=12Hz), 4.70(1H, t, JL32 8 Hz), 5.0(1H, d, J=5 Hz), 
5.55(1H, dd, J=5 & 8Hz), 7.20(1H, broad), 7.60 (1H, broad), 7.86(4H, s), 
8.74(1H, d, J=8Hz) 
EXAMPLE 28 
In water (100 ml) is dissolved 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-[2-(N-carboethoxycarbamoyl 
)benzoyloxy]methyl-3-cephem-4-carboxylic acid (3.62 g) together with 
2-(2-hydroxyethylthio)-5-mercapto-1,3,4-thiadiazole (1.45 g) and sodium 
bicarbonate (1.1 g). The solution is adjusted to pH 5.5 and reacted at 
60.degree. C. for 50 minutes. The aqueous solution is washed twice with 
ethyl acetate, brought down to pH 2.0 with diluted hydrochloric acid and 
extracted three times with a mixture of ethyl acetate and tetrahydrofuran 
(4:1). The organic layer is washed with a saturated aqueous solution of 
sodium chloride, dried over magnesium sulfate, filtered and distilled off 
the solvent. The procedure provides 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-[2-(2-hydroxyethylthio)-1, 
3,4-thiadiazol-5-yl]-thiomethyl-3-cephem-4-carboxylic acid (3.15 g). 
IR(KBr): cm.sup.-1 3325, 1780, 1715, 1645, 1530 
NMR(d.sub.6 -DMSO): .delta. 1.30-2.40(6H, m), 3.20-3.80(6H, m), 4.27(2H, 
ABq, J=12 Hz), 4.65(1H, t, J=9 Hz), 4.96(1H, d, J=5 Hz), 5.55(1H, dd, J=5 
& 8Hz), 7.87(4H, s), 8.70(1H, d, J=8 Hz) 
EXAMPLE 29 
7.beta.-(2-Thienylacetamido)-3-[2-(N-carboethoxysulfamoyl)-benzoyloxy]meth 
yl-3-cephem-4-carboxylic acid (619 mg) is dissolved in water (5 ml) 
together with sodium hydrogen carbonate (170 mg), potassium iodide (400 
mg) and pyridine (210 mg). The solution is adjusted to pH 6.5 and then 
stirred at 60.degree. C. for one hour and a half. After cooling, the 
reaction mixture is subjected to column chromatography on Amberlite XAD-2. 
Elution is carried out with water and then, with a mixture of 
water-methanol. The fractions containing the desired product are pooled, 
concentrated and lyophilized. The procedure provides 
7.beta.-(2-thienylacetamido)-3-(pyridiniummethyl)-3-cephem-4-carboxylate 
(220 mg). 
IR(KBr): cm.sup.-1 1763, 1698, 1617 
NMR(D.sub.2 O): .delta. 3.17 & 3.67(2H, ABq, J=17 Hz, 2--CH.sub.2), 
3.38(2H, s, --CH.sub.2 CO--), 5.19(1H, d, J=5 Hz, 6--H), 5.41 & 5.67(2H, 
ABq, J=14 Hz, 3--CH.sub.2), 5.75(1H, d, J=5 Hz, 7--H), 
##STR20## 
EXAMPLE 30 
7.beta.-Amino-3-[2-(N-carboethoxycarbamoyl)benzoyloxy]-methyl-3-cephem-4-ca 
rboxylic acid (450 mg) is suspended in N,N-dimethylformamide (5 ml), 
followed by addition of O-carboxymandelic anhydride (267 mg). The mixture 
is stirred for 2 hours and poured into the mixed solution of water (20 ml) 
and ethyl acetate (20 ml). After separation, the ethyl acetate layer is 
washed with water and then extracted with aqueous sodium bicarbonate 
solution. The aqueous layer is purified by column chromatography on 
Sephadex LH-20. The main fraction is collected and lyophilized. The 
procedure provides sodium 
7.beta.-mandelamido-3-[2-(N-carboethoxycarbamoyl)benzoyloxy]methyl-3-cephe 
m-4-carboxylate (180 mg). 
IR(KBr): cm.sup.-1 1770, 1505 
NMR(d.sub.6 -DMSO): .delta. 1.17(3H, t, --CH.sub.2 CH.sub.3), 3.47(2H, 
2--CH.sub.2), 4.08 (2H, q, --CH.sub.2 CH.sub.3), 4.9-5.4(4H, m), 5.59(1H, 
dd, 7-H), 7.2-8.0(9H, m) 
EXAMPLE 31 
7.beta.-Amino-3-[2-(N-carboethoxysulfamoyl)benzoyloxy]-methyl-3-cephem-4-ca 
rboxylic acid (970 mg) is suspended in dichloromethane (20 ml), followed by 
the addition of triethylamine (0.84 ml) and D(-)-.alpha.-sulfophenylacetyl 
chloride (560 mg) under ice-cooling. The mixture is stirred for 30 minutes 
and after the addition of more triethylamine (0.2 ml), stirred further for 
30 minutes. The solvent is distilled off to dryness and the residue is 
triturated with ether (60 ml). Thus obtained powder is collected by 
filtration, washed with ether and dried over phosphorus pentoxide. The 
powder is dissolved in water (8 ml) and under ice-cooling 1N-hydrochloric 
acid is added to adjust its pH to 1.0 and then a saturated aqueous 
solution of sodium chloride (12 ml) is added. The resultant precipitate is 
triturated, collected by filtration, washed with a saturated aqueous 
solution of sodium chloride and dried. Thus obtained powder is dissolved 
in tetrahydrofuran (40 ml) and the insolubles are filtered off. The 
filtrate is concentrated to yield slurry. The slurry is triturated with 
ether (40 ml) and the resultant precipitate is collected by filtration and 
dried over phosphorus pentoxide. The procedure provides 
7.beta.-D-.alpha.-sulfophenylacetamido)-3-[2-(N-carboethoxy-sulfamoyl)benz 
oyloxy]methyl-3-cephem-4-carboxylic acid (822 mg). 
IR(KBr): cm.sup.-1 1780(sh), 1745, 1680 
NMR(d.sub.6 -DMSO): .delta. 1.11(3H, t, J=8 Hz --CH.sub.3), 3.60(2H, broad, 
2--CH.sub.2), 4.03(2H, q, J=8 Hz, --CH.sub.2 CH.sub.3), 5.07(1H, s, 
##STR21## 
5.09 (1H, d, J=4.5 Hz, 6--H), 5.18(2H, ABq, J=13Hz, 3--CH.sub.2), 5.78 
(1H, q, J=4.5 & 8Hz, 7--H), 7.6-8.15(9H, m, arom-H) 
EXAMPLE 32 
In acetone (1 ml) is dissolved 
7.beta.-(4-bromo-3-oxobutylamido)-3-[2-(N-carboethoxysulfamoyl)benzoyloxy] 
methyl-3-cephem-4-carboxylic acid (319 mg) and, under ice-cooling, water (1 
ml) and thiourea (42 mg) is added. Then, sodium hydrogen carbonate (42 mg) 
and water (1 ml) are added. The mixture is reacted at room temperature for 
5 hours, after which time it is cooled with ice. The resultant crystals 
are collected by filtration, washed with water and ether and dried. The 
procedure provides 
7.beta.-(2-imino-4-thiazolin-4-yl)acetamido-3-[2-(N-carboethoxysulfamoyl)b 
enzoyloxy]methyl-3-cephem-4-carboxylic acid. 
NMR(d.sub.6 -DMSO): .delta. 1.10(3H, t, CH.sub.2 CH.sub.3), 3.38(2H, s, 
--CH.sub.2 CONH), 3.58(2H, broad, 2--CH.sub.2), 4.01(2H, q, CH.sub.2 
CH.sub.3), 5.07(1H, d, J=5 Hz, 6--H), 5.17(2H, ABq, J=13 Hz, 3--CH.sub.2), 
5.70(1H, dd, J=5 & 8Hz, 7--H), 6.23(1H, s, thiazolin-H) 
EXAMPLE 33 
In 50% aqueous acetone (12 ml), is dissolved 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-[2-(N-carboethoxycarbamoyl 
)benzoyloxy]methyl-3-cephem-4-carboxylic acid (723 mg), followed by the 
addition of indole (0.35 g) and sodium hydrogen carbonate (0.17 g). The 
mixture is stirred at 60.degree. C. for one hour. After cooling, most of 
the acetone is distilled off under reduced pressure. Then 5% aqueous 
phosphoric acid solution (15 ml) and ethyl acetate (30 ml) are added. The 
ethyl acetate layer is washed with a saturated aqueous solution of sodium 
chloride, dried and concentrated. To the residue, ether is added and the 
resultant powder is collected by filtration. This powder is suspended in 
water and dissolved by the addition of sodium hydrogen carbonate (0.17 g). 
The solution is subjected to column chromatography on Sephadex LH-20, 
elution being carried out with water. The fractions containing the desired 
product are pooled and lyophilized. The procedure provides 
7.beta.-(D-5-phthalimido-5-carboxyvaleramido)-3-(3-indolyl)methyl-3-cephem 
-4-carboxylic acid disodium salt (0.24 g). 
IR(KBr): cm.sup.-1 1758, 1702, 1600 
NMR(D.sub.2 O): .delta. 1.68 & 2.26(6H, --(CH.sub.2).sub.3 --), 2.45 & 
2.82(2H, ABq, J=18 Hz, 2--CH.sub.2), 3.61 & 3.86(2H, ABq, J=15 Hz, 
3--CH.sub.2), 4.6-4.9(2H, --CH-- & 6--H), 5.46(1H, d, J=5 Hz, 7--H), 
7.0-7.8 (10H, 
##STR22## 
EXAMPLE 34 
In 50% aqueous acetone (8 ml) is dissolved 
7.beta.-thienylacetamido-3-[2-(N-carboethoxysulfamoyl)benzoyloxy]methyl-3- 
cephem-4-carboxylic acid-etherate (619 mg), together with acetylacetone 
(0.5 g) and sodium hydrogen carbonate (0.17 g). The mixture is stirred at 
60.degree. C. for one hour. After cooling, most of the acetone is 
distilled off under reduced pressure. To the residue, ethyl acetate (20 
ml) and diluted phosphoric acid are added. The ethyl acetate layer is 
washed with a saturated aqueous solution of sodium chloride, dried over 
magnesium sulfate and concentrated. To the residue, ether is added and the 
resultant powder is collected by filtration. This powder is suspended in 
water and dissolved by the addition of sodium hydrogen carbonate. The 
solution is subjected to column chromatography on Sephadex LH-20, elution 
being carried out with water. The desired fractions are pooled and 
lyophilized. This product is dissolved in water and acidified with diluted 
phosphoric acid and extracted with ethyl acetate (20 ml). The ethyl 
acetate layer is washed with water, dried over magnesium sulfate and 
concentrated under reduced pressure. To the residue, ether is added and 
triturated. The resultant powder is collected by filtration, washed with 
ether and dried. The procedure provides 
7.beta.-thienylacetamido-3-(2-acetyl-3-oxo)butyl-3-cephem-4-carboxylic 
acid. 
IR(KBr): cm.sup.-1 1765, 1718 
NMR(d.sub.6 -DMSO): .delta. 2.13 & 2.18(6H, s, (COCH.sub.3).sub.2), 
2.6-3.1(2H, m, 3--CH.sub.2), 3.31 & 3.56(2H, ABq, 2--CH.sub.2), 3.75(2H, 
s, --CH.sub.2 CO--), 4.14(1H, --CH--), 5.01(1H, d, J=5 Hz, 6--H), 5.58(1H, 
dd, J=5 & 8 Hz, 7--H), 
##STR23## 
9.04(1H, d, J=8 Hz, --CONH--) 
EXAMPLE 35 
(1) Sodium 
7.beta.-(t-butoxycarbonyl)amino-3-hydroxymethyl-3-cephem-4-carboxylate 
(1.60 g) is dissolved in N,N-dimethylformamide (8 ml), followed by 
addition of triethylamine (2 ml) and N-carboethoxyphthalimide (1.65 g) 
under stirring and ice-cooling. After stirring for one hour at room 
temperature, the solution is poured into toluene (300 ml) and n-hexane 
(200 ml) is added. The resultant precipitate is collected by filtration, 
washed with toluene and dried. The powder thus obtained is dissolved in 
water (50 ml) under ice-cooling and ethyl acetate (50 ml) is added. 
Diluted phosphoric acid is added to adjust its pH to 4.0. After 
separation, the ethyl acetate layer is washed with water and water (10 ml) 
is added. To the mixture, sodium hydrogen carbonate is added to adjust its 
pH to 6.8. 
The aqueous layer is washed with ethyl acetate and ethyl acetate (50 ml) is 
added. The mixture is treated at pH 4.0 in a similar way just mentioned 
above. 
After separation, the ethyl acetate layer is washed with water, dried over 
anhydrous sodium sulfate and concentrated. To the residue, ether (70 ml) 
is added under stirring and the resultant insolubles are filtered off. The 
filtrate is concentrated (to 3 ml) under reduced pressure and carbon 
tetrachloride (50 ml) is added. The resultant precipitate is collected by 
filtration, washed with carbon tetrachloride and dried over phosphorus 
pentoxide. The procedure provides 
7.beta.-(t-butoxycarbonyl)-amino-3-[2-(N-carboethoxycarbamoyl)benzoyloxy]m 
ethyl-3-cephem-4-carboxylic acid (1.04 g). 
IR(KBr): cm.sup.-1 1775, 1716 
NMR(d.sub.6 -DMSO): .delta. 1.18(3H, t, J=8 Hz, --CH.sub.2 CH.sub.3), 
1.44(9H, s, C(CH.sub.3).sub.3), 3.61(2H, ABq, J=18 Hz, 2--CH.sub.2), 
4.07(2H, q, J=8Hz, --CH.sub.2 CH.sub.3), 5.08(1H, d, J=4.5, 6--H), 
5.10(2H, ABq, J=13Hz, 3--CH.sub.2), 5.44(1H, q, J=4.5 & 8Hz, 7--H), 
7.30-7.96(4H, m, arom-H) 
(2) 
7.beta.-(t-Butoxycarbonyl)amino-3-[2-(N-carboethoxycarbamoyl)benzoyloxy]me 
thyl-3-cephem-4-carboxylic acid (275 mg) is dissolved in trifluoroacetic 
acid (2 ml) under ice-cooling and the mixture is stirred for 20 minutes. 
The solution is concentrated and to the residue, ether (20 ml) is added. 
The resultant precipitate is triturated, collected by filtration, washed 
with ether and dried over phosphorus pentoxide. The procedure provides 
7.beta.-amino-3-[2-(N-carboethoxycarbamoyl)benzoyloxy]methyl-3-cephem-4-ca 
rboxylic acid (195 mg). In NMR spectrum, this product is good agreement 
with the product obtained in Example 20. 
EXAMPLE 36 
(1) In N,N-dimethylformamide (8 ml), is dissolved sodium 
7.beta.-(t-butoxycarbonyl)amino-3-hydroxymethyl-3-cephem-4-carboxylate 
(1.60 g), followed by addition of triethylamine (3 ml) and 
N-tosylphthalimide (1.80 g) under ice-cooling. The mixture is stirred for 
one hour and then poured into toluene (300 ml). n-Hexane (200 ml) is added 
and the resultant precipitate is collected by filtration, washed with 
toluene and dried. The powder thus obtained is dissolved in water (50 ml). 
The solution is adjusted to pH 3.7 with acetic acid and a saturated 
aqueous solution of sodium chloride (50 ml) is added. The resultant 
precipitate is collected by filtration and washed with water. The cake 
thus obtained is added to a mixed solution of ethyl acetate (30 ml) and 
water (10 ml). After separation, the ethyl acetate layer is washed with 
water and water (10 ml) is added. Sodium hydrogen carbonate is added to 
adjust to pH 6.8. After separation, the aqueous layer is washed with ethyl 
acetate and ethyl acetate (20 ml) is added. To the mixture, diluted 
phosphoric acid is added to adjust to pH 4.0. After separation, the ethyl 
acetate layer is washed with water, dried over anhydrous sodium sulfate 
and concentrated. To the residue, ether is added under stirring and the 
resultant insolubles are filtered off. The filtrate is concentrated under 
reduced pressure and carbon tetrachloride is added. The resultant 
precipitate is collected by filtration and dried over phosphorous 
pentoxide. The procedure provides 
7.beta.-(t-butoxycarbonyl)amino-3-[2-(N-tosylcarbamoyl)benzoyloxy]methyl-3 
-cephem-4-carboxylic acid (1.21 g). 
IR(KBr): cm.sup.-1 1788, 1715 
NMR(d.sub.6 -DMSO): .delta.1.44(9H, s, --C(CH.sub.3).sub.3), 2.42(3H, s, 
--CH.sub.3), 3.48 (2H, ABq, J=18 Hz, 2--CH.sub.2), 4.93(2H, ABq, J=13 Hz, 
3--CH.sub.2), 5.04(1H, d, J=5 Hz, 6--H), 5.48(1H, q, J=5 & 8 Hz, 7--H), 
7.40-7.93(8H, m, arom-H) 
(2) 
7.beta.-(t-Butoxycarbonyl)amino-3-[2-(N-tosylcarbamoyl)benzoyloxy]methyl-3 
-cephem-4-carboxylic acid (631 mg) is dissolved in trifluoroacetic acid (2 
ml) under ice-cooling. The solution is stirred for 20 minutes and 
concentrated under reduced pressure. To the residue, ether (20 ml) is 
added and the resultant solid is triturated. The powder thus obtained is 
collected by filtration, washed with ether and dried over phosphorus 
pentoxide. The procedure provides 
7.beta.-amino-3-[2-(N-tosylcarbamoyl)benzoyloxy]methyl-3-cephem-4-carboxyl 
ic acid (480 mg). 
IR(KBr): cm.sup.-1 1782, 1715, 1680 
NMR(D.sub.2 O+NaHCO.sub.3): .delta.2.54(3H, s, --CH.sub.3), 3.81(2H, ABq, 
J=18 Hz, 2--CH.sub.2), 4.56(2H, ABq, J=12 Hz, 3--CH.sub.2), 4.86(1H, d, 
J=5 Hz, 6--H), 5.16(1H, d, J=5 Hz, 7--H), 7.41-8.02(8H, m, arom-H) 
EXAMPLE 37 
7.beta.-Amino-3-[2-(N-carboethoxysulfamoyl)benzoyloxy]methyl-3-cephem-4-car 
boxylic acid (457 mg) is suspended in a mixed solution of dichloromethane 
(5 ml) and N,N-dimethylformamide (1.0 ml) and to the mixture, a solution 
of (1H-tetrazol-1-yl)acetyl chloride (161 mg) in tetrahydrofuran (2 ml) is 
added over a period of 20 minutes and stirred for 2 hours. The solvent is 
distilled off under reduced pressure and to the resultant viscous residue, 
ethyl acetate (50 ml) and water (20 ml) are added and stirred. After 
separation, the ethyl acetate layer is washed with water, dried over 
anhydrous sodium sulfate and concentrated. To the concentrate (5 ml), 
chloroform (50 ml) is added and the resultant precipitate is filtered off. 
The filtrate is concentrated and to the concentrate (5 ml), ether (50 ml) 
is added. The resultant precipitate is collected by filtration and dried 
over phosphorous pentoxide. The procedure provides 
7.beta.-(1H-tetrazol-1-yl)acetamido-3-[2-(N-carboethoxysulfamoyl)benzoylox 
y]methyl-3-cephem-4-carboxylic acid (216 mg). 
IR(KBr): cm.sup.-1 1782, 1745 
NMR(d.sub.6 -DMSO): .delta.1.13(3H, t, J=7 Hz, --CH.sub.2 CH.sub.3), 
3.70(2H, broad, 2--CH.sub.2), 4.05(2H, q, J=7 Hz, --CH.sub.2 CH.sub.3), 
5.15(1H, d, J=4.5 Hz, 6--H), 5.22(2H, ABq, J=13.5 Hz, 3--CH.sub.2), 
5.40(2H, s, --N--CH.sub.2), 
5.78(1H, q, J=4.5 & 8 Hz, 7--H), 7.64-8.14(4H, m, arom-H), 9.35(1H, s, 
tetrazole-H), 9.49(1H, d, J=8 Hz, 7--CONH--) 
EXAMPLE 38 
In water (5 ml) are dissolved 
7.beta.-(2-imino-4-thiazolin-4-yl)acetamido-3-[2-(N-carboethoxysulfamoyl)b 
enzoyloxy]methyl-3-cephem-4-carboxylic acid (313 mg), 
5-mercapto-1-[2-(N,N-dimethylamino)ethyl]-1H-tetrazole (113 mg) and sodium 
hydrogen carbonate (42 mg) and the resultant solution is heated at 
55.degree. C. for 60 minutes. The reaction solution is adjusted its pH to 
5.8 and purified by column chromatography on Amberlite XAD-2 and the 
fractions rich in the desired product are pooled and lyophilized. The 
procedure provides 
7.beta.-(2-imino-4-thiazolin-4-yl)acetamido-3-{1-[2-(N,N-dimethylamino)eth 
yl]-1H-tetrazol-5-yl}thiomethyl-3-cephem-4-carboxylic acid. 
IR(KBr): cm.sup.-1 1765 
NMR(D.sub.2 O): .delta.3.06(6H, s, --N(CH.sub.3).sub.2), 3.5-4.8(10H, m), 
5.12(1H, d, J=5 Hz, 6--H), 5.65(1H, d, J=5 Hz, 7--H), 6.62(1H, s, 
thiazolin-H) 
EXAMPLE 39 
7.beta.-(D-.alpha.-Sulfophenylacetamido)-3-[2-(N-carboethoxysulfamoyl)benzo 
yloxy]methyl-3-cephem-4-carboxylic acid (341 mg) is dissolved in water (0.3 
ml) by the addition of sodium hydrogen carbonate (84 mg) and then 
isonicotinamide (90 mg) and KSCN (1.2 g) are added. The mixture is heated 
at 60.degree. C. for one hour and then subjected to column chromatography 
on Amberlite XAD-2, elution being carried out with water. The fractions 
rich in the desired product are pooled, lyophilized and recrystallized 
from ethanol-water. The procedure provides 
7.beta.-(D-.alpha.-sulfophenylacetamido)-3-(4-carbamoylpyridinium)methyl-3 
-cephem-4-carboxylate sodium salt. 
IR(KBr): cm.sup.-1 1765, 1692, 1645, 1615, 1029 
NMR(D.sub.2 O): .delta.2.99 & 3.56(2H, ABq, J=18 Hz, 2--CH.sub.2), 5.40 & 
5.51 (2H, 3--CH.sub.2), 5.13(1H, d, J=4.8, 6--H), 5.73(1H, d, J=4.8 Hz, 
7--H), 
##STR24## 
7.40(5H, m), 8.31 & 9.07(4H) 
EXAMPLE 40 
In a phosphate buffer solution of pH 6.4 (3 ml) are dissolved 
5-mercapto-2-methyl-1,3,4-thiadiazole (79 mg), sodium hydrogen carbonate 
(92 mg) and 
7.beta.-(1H-tetrazol-1-yl)acetamido-3-[2-(N-carboethoxysulfamoyl)benzoylox 
y]methyl-3-cephem-4-carboxylic acid (298 mg) and the resultant solution is 
heated at 60.degree. C. for one hour. Then, the mixture is concentrated 
and the residue is subjected to column chromatography on Sephadex LH-20, 
elution being carried out with water. The fractions containing the desired 
product are pooled and lyophilized. The procedure provides sodium 
7.beta.-(1H-tetrazol-1-yl)acetamido-3-(2-methyl-1,3,4-thiadiazol-5-yl)thio 
methyl-3-cephem-4-carboxylate. This product is in good agreement with the 
authentic sample in NMR spectrum.