7.beta.-[D-2-Amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-Hal-3-cephem-4-ca rboxylic acids, wherein Hal denotes halogen with an atomic number of up to 35, and pharmaceutically acceptable salts thereof are prepared, which compounds have antibiotic activity.

The invention relates to 
7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-Hal-3-cephem-4-c 
arboxylic acids (I), wherein Hal denotes halogen with an atomic number of 
up to 35, or salts thereof, and to processes for their manufacture and 
also to pharmaceutical formulations containing these compounds as the 
active compounds, and to their use, preferably in the form of 
pharmaceutical formulations. 
In the abovementioned compounds Hal represents fluorine or bromine but 
above all represents chlorine. 
Salts are, in particular, non-toxic salts which can be used 
pharmaceutically, such as metal salts or ammonium salts, such as alkali 
metal salts and alkaline earth metal salts, for example sodium salts, 
potassium salts, magnesium salts or calcium salts, and also ammonium salts 
with ammonia or suitable organic amines, and, above all, aliphatic, 
cycloaliphatic, cycloaliphatic-aliphatic and araliphatic primary, 
secondary or tertiary mono-, di- or poly-amines, and also heterocyclic 
bases, can be used for forming the salts, such as lower alkylamines, for 
example triethylamine, hydroxy-lower alkylamines, for example 
2-hydroxyethylamine, bis-(2-hydroxyethyl)-amine or 
tri-(2-hydroxyethyl)-amine, basic aliphatic esters of carboxylic acids, 
for example 2-diethylamino-ethyl 4-aminobenzoate, lower alkyleneamines, 
for example 1-ethyl-piperidine, cycloalkylamines, for example 
bicyclohexylamine, or benzylamines, for example 
N,N'-dibenzyl-ethylenediamine, and also bases of the pyridine type, for 
example pyridine, collidine or quinoline. The new compounds can also form 
acid addition salts, for example with inorganic acids, such as 
hydrochloric acid, sulphuric acid or phosphoric acid, or with suitable 
organic carboxylic or sulphonic acids, for example trifluoroacetic acid or 
4-methylphenylsulphonic acid. The new compounds are preferably in the form 
of their inner salts, that is to say in the form of the zwitter-ion. 
The new compounds of the present invention display valuable antibiotic 
properties. Thus, in the free form or in the form of their salts, they are 
active in vitro, in doses of 0.1 to 100 mcg/ml against cocci, for example 
Staphylococcus aureus, Staphylococcus faecalis, Diplococcus pneumoniae, 
Neisseria meningitidis and Neisseria gonorrhoeae and, in doses of 0.4 to 
50 mcg/ml, against enterobacteriaceae, for example Escherichia coli, 
Klebsiella pneumoniae, Salmonella typhimurium and Proteus mirabilis. 
When administered parenterally or, in particular, orally, they are active 
against micro-organisms, such as Gram-positive bacteria, for example 
Staphylococcus aureus, Streptococcus pyogenes and Diplococcus pneumoniae 
(for example in mice in doses of about 0.0025 to about 0.04 g/kg per os.) 
and Gram-negative bacteria, for example Escherichia coli, Salmonella 
typhimurium, Shigella flexneri, Klebsiella pneumoniae, Proteus vulgaris, 
Proteus rettgeri and Proteus mirabilis (for example in mice in doses of 
about 0.008 to about 0.03 g/kg per os.) and especially against 
penicillin-resistant bacteria, this action being coupled with low 
toxicity. These new compounds can therefore be used, for example in the 
form of formulations having an antibiotic action, for the treatment of 
corresponding infections. 
The invention relates above all to 
7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-chloro-3-cephem- 
4-carboxylic acid and its salts, especially its non-toxic salts which can 
be used pharmaceutically and in particular its inner salt. Compared with 
7.beta.-(D-2-amino-2-phenyl-acetylamino)-3-chloro-3-cephem-4-carboxylic 
acid, which is known from U.S. Pat. No. 3,925,372, this compound is 
distinguished by a greater stability at the physiological pH value. 
The new compounds are manufactured in a manner which is in itself known. 
Thus, for example, they can be obtained when the 3-hydroxyl group in a 
7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-hydroxy-3-cephem 
-4-carboxylic acid (II), wherein the carboxyl group is in a protected form 
and the amino group is preferably in a protected form, is converted into a 
halogen atom Hal and, if necessary, in a resulting compound, a protected 
amino group which may be present is converted into the free amino group 
and/or the protected carboxyl group is converted into the free carboxyl 
group and, if desired, a resulting salt is converted into the free 
compound or into another salt and/or a free compound is converted into a 
salt. 
In the above starting material the carboxyl group is in a protected form 
and the amino group is usually in a protected form, possible protective 
groups being those used in penicillin and cephalosporin chemistry and in 
peptide chemistry. 
The carboxyl group is usually protected in the esterified form and an ester 
grouping of this type can be split easily under gentle conditions. 
Possible suitable protected carboxyl groups are, in particular, lower 
alkoxycarbonyl, especially tert.-lower alkoxycarbonyl, for example 
tert.-butoxycarbonyl, polycycloalkoxycarbonyl, for example 
adamantyloxycarbonyl, arylmethoxycarbonyl, wherein aryl preferably 
represents one or two phenyl radicals which are optionally monosubstituted 
or polysubstituted, for example by lower alkyl, especially tert.-lower 
alkyl, for example tert.-butyl, lower alkoxy, such as methoxy, hydroxyl, 
halogen, for example chlorine, and/or nitro, such as optionally 
substituted benzyloxycarbonyl, for example benzyloxycarbonyl substituted 
as indicated above, for example 4-nitro-benzyloxycarbonyl, or 
diphenylmethoxycarbonyl which, for example, is substituted as indicated 
above, for example benzhydryloxycarbonyl or 
di-(4-methoxyphenyl)-methoxycarbonyl, or 2-halogeno-lower alkoxycarbonyl, 
for example 2,2,2-trichloroethoxycarbonyl or 2-bromo- or 
2-iodo-ethoxycarbonyl, or acylmethoxycarbonyl, especially 
aroylmethoxycarbonyl, wherein the aroyl group preferably represents benzyl 
which is optionally substituted, for example by halogen, such as bromine, 
for example phenacyloxycarbonyl. Esterified carboxyl groups are also 
corresponding silyloxycarbonyl groups, especially organic silyloxycarbonyl 
groups. In these groups, the silicon atom preferably has lower alkyl, 
especially methyl, and lower alkoxy, for example methoxy, and/or halogen, 
for example chlorine, as substituents. Suitable silyl protective groups 
are, above all, tri-lower alkylsilyl, especially trimethylsilyl, and also 
dimethyl-tert.-butyl-silyl, lower alkoxy-lower alkyl-halogeno-silyl, for 
example methoxy-methyl-chloro-silyl, or di-lower alkyl-halogeno-silyl, for 
example dimethyl-chloro-silyl. It is also possible for silyl protective 
groups, and especially those which contain a halogen atom as a 
substituent, at the same time to protect the carboxyl groups in two 
different molecules of the starting material; that is to say in such 
groups the halogen atom has been replaced by the carboxyl group of a 
further molecule of the starting material. 
A preferred protected carboxyl group is, in particular, optionally 
substituted benzyloxycarbonyl, for example benzyloxycarbonyl which is 
substituted as mentioned above, for example 4-nitrobenzyloxycarbonyl, or 
optionally substituted diphenylmethoxycarbonyl, for example 
diphenylmethoxycarbonyl which is substituted as mentioned above, for 
example benzhydroloxycarbonyl. 
A protected amino group can be, for example, in the form of an acylamino, 
triarylmethylamino, etherified mercaptoamino, 1-acyl-2-lower 
alkylideneamino or silylamino group which can be split easily. 
In a corresponding acylamino group, acyl is preferably the acyl radical of 
a carbonic acid half-ester, such as lower alkoxycarbonyl, especially 
tert.-lower alkoxycarbonyl, for example tert.-butoxycarbonyl, 
polycycloalkoxycarbonyl, for example adamantyloxycarbonyl, 
arylmethoxycarbonyl, wherein aryl preferably represents one or two phenyl 
radicals which are optionally monosubstituted or polysubstituted, for 
example by lower alkyl, especially tert.-lower alkyl, for example 
tert.-butyl, lower alkoxy, such as methoxy, hydroxyl, halogen, for example 
chlorine, and/or nitro, such as optionally substituted benzyloxycarbonyl, 
for example benzyloxycarbonyl which is substituted as mentioned above, for 
example 4-nitrobenzyloxycarbonyl, or diphenylmethoxycarbonyl which is 
substituted, for example as indicated above, for example 
benzhydryloxycarbonyl or di-(4-methoxyphenyl)-methoxycarbonyl, or 
2-halogeno-lower alkoxycarbonyl, for example 2,2,2-trichloroethoxycarbonyl 
or 2-iodoethoxycarbonyl, or acylmethoxycarbonyl, especially 
aroylmethoxycarbonyl wherein the aroyl group preferably represents 
optionally substituted benzoyl, for example benzoyl substituted by 
halogen, such as bromine, for example phenacyloxycarbonyl. In an acylamino 
group, acyl can also represent the corresponding radical of an organic 
sulphonic acid; such a radical is, in particular, arylsulphonyl wherein 
aryl denotes a phenyl radical which is optionally substituted, for example 
by lower alkyl, such as methyl, halogen, such as bromine, or nitro, for 
example 4-methylphenylsulphonyl. 
In a triarylmethylamino group, the aryl radicals are, in particular, 
optionally substituted phenyl radicals; a corresponding group is, above 
all, trityl. 
An etherified mercapto group in an amino group protected by such a radical 
is, above all, arylthio or aryl-lower alkylthio, wherein aryl is, in 
particular, phenyl which is optionally substituted, for example by lower 
alkyl, such as methyl or tert.-butyl, lower alkoxy, such as methoxy, 
halogen, such as chlorine, and/or nitro. A corresponding amino protective 
group is, for example, 4-nitrophenylthio. 
In a 1-acyl-2-lower alkylidene radical which can be used as an amino 
protective group, acyl is preferably the corresponding radical of a lower 
alkanecarboxylic acid, of a benzoic acid which is optionally substituted, 
for example by lower alkyl, such as methyl or tert.-butyl, lower alkoxy, 
such as methoxy, halogen, such as chlorine, and/or nitro, or of a carbonic 
acid half-ester, such as of a carbonic acid lower alkyl half-ester. 
Corresponding protective groups are, above all, 1-lower 
alkanoyl-2-propylidene, for example 1-acetyl-2-propylidene, or 1-lower 
alkoxycarbonyl-2-propylidene, for example 1-ethoxycarbonyl-2-propylidene. 
A silylamino group is, above all, an organic silylamino group wherein the 
silicon atom preferably has lower alkyl, especially methyl, and also lower 
alkoxy, for example methoxy, and/or halogen, for example chlorine, as 
substituents. Corresponding silyl groups are, above all, tri-lower 
alkylsilyl, especially trimethylsilyl, and also dimethyl-tert.-butylsilyl, 
lower alkoxy-lower alkyl-halogeno-silyl, for example 
methoxy-methyl-chlorosilyl, or di-lower alkyl-halogeno-silyl, for example 
dimethyl-chlorosilyl. It is possible for silyl protective groups, 
especially those which contain a halogen atom as a substituent, at the 
same time to protect the amino group in two different molecules of the 
starting material; that is to say in such groups the halogen atom has been 
replaced by the amino group of a further molecule of the starting 
material. 
The amino group in the starting material II can also be protected in a 
protonised form; possible anions are, above all, those of strong inorganic 
acids, such as hydrogen halide acids, for example the chlorine or bromine 
anion. 
Preferred amino protective groups are the acyl radicals of carbonic acid 
half-esters, especially tert.-lower alkoxycarbonyl, benzyloxycarbonyl or 
diphenylmethoxycarbonyl, which are optionally substituted, for example as 
indicated, or 2-halogeno-lower alkoxycarbonyl. 
The starting material II is preferably in the indicated 3-hydroxy-3-cephem 
form but can also be employed in the corresponding tautomeric cepham-3-one 
form. 
The replacement of the hydroxyl group by halogen can be carried out in 
various ways, usually by treatment with a halogenating, that is to say 
fluorinating, chlorinating or brominating, agent. 
Compounds I wherein Hal denotes fluorine, chlorine or bromine can be 
manufactured, for example, by treating a starting material II with a 
phosphorus reagent which replaces enol-hydroxyl groups by halogen and 
subsequently splitting off the protective groups which are present. 
Phosphorus reagents of this type are, for example, 
dihalogeno-triorganyl-phosphoranes, trihalogeno-diorganyl-phosphoranes or 
a mixture consisting of a triorganyl-phosphine and a carbon tetrahalide. 
In these reagents halogen is fluorine, chlorine or bromine. In the carbon 
tetrahalide, halogen is preferably chlorine or bromine. The organyl 
radicals in the phosphoranes and phosphines are organic radicals with up 
to 18 carbon atoms and can be identical or different. 
Organyl radicals are, in particular, hydrocarbon radicals which have up to 
18, especially up to 12 and preferably up to 6, carbon atoms and are 
optionally substituted, for example by tertiary amino groups or polymers, 
such as lower alkyl radicals, for example methyl, ethyl or propyl, 
di-lower alkylamino-lower alkyl radicals, for example 
3-dimethylaminopropyl, carbocyclic radicals, such as phenyl which is 
optionally substituted as indicated, and also phenyl which is substituted 
by polymers, for example by polystyrene cross-linked with divinylbenzene, 
or phenyl which is substituted by di-lower alkylamino-lower alkyl, for 
example dimethylaminomethyl. In the case of phenyl substituted by a 
polymer there is usually only one radical present on a given phosphorus 
atom. 
Further organyl radicals are secondary amino radicals, such as di-lower 
alkylamino, above all dimethylamino. 
Representative examples of the phosphoranes mentioned are 
difluoro-triphenyl-, trifluoro-diphenyl-, dichloro-triphenyl-, 
trichloro-diphenyl-, dibromo-triphenyl- and tribromodiphenyl-phosphorane, 
wherein one of the phenyl groups can be substituted by a polymer, such as 
a polystyrene crosslinked with divinylbenzene, or by dimethylaminomethyl. 
Representative examples of the phosphines mentioned are triethyl-, 
methyl-propyl-phenyl-, bis-(3-dimethylaminopropyl)-phenyl-, 
tris-(dimethylamino)-, bis-(dimethylamino)-phenyl- and, in particular, 
triphenyl-phosphine, wherein one of the phenyl groups can be substituted 
by a polymer, such as a polystyrene crosslinked with divinylbenzene. 
Carbon tetrahalides are, for example, carbon tetrabromide and, in 
particular, carbon tetrachloride. 
The reaction with the halogenating phosphorus reagents takes place in a 
manner which is in itself known in an inert aprotic, preferably polar, 
solvent, such as a chlorinated hydrocarbon, for example methylene 
chloride, chloroform, carbon tetrachloride of 1,2-dichloroethane, a 
nitrile, such as acetonitrile or benzonitrile, or a N,N-disubstituted 
carboxylic acid amide, such as dimethylformamide or N,N-dimethylacetamide, 
or mixtures thereof, and, depending on the reactivity of the reagent 
employed, with cooling or warming, that is to say at temperatures between 
about -60.degree. C and the reflux temperature of the solvent used, and 
optionally in an inert gas atmosphere, such as a nitrogen atmosphere. When 
tri-lower alkyl-phosphines or tris-(di-lower alkylamino)phosphines and 
carbon tetrachloride or carbon tetrabromide are used, cooling is usually 
necessary, say to -60.degree. to -20.degree. C. 
The halogenating phosphoranes mentioned can also be formed in situ, for 
example by reacting the said phosphines with the desired carbon 
tetrahalide, in which case other halogenating phosphorus compounds are 
also formed in addition to the dihalogeno-triorganyl phosphorane, or by 
treating the phosphines with a halogen, for example chlorine, or by 
reacting triorganylphosphine oxides with a dihalogenocarbonyl, such as 
phosgene, or trihalogenosilane, such as trichlorosilane. 
When halogenating with the said phosphoranes, a weak base, such as pyridine 
or a N,N-di-lower alkylaniline, such as N,N-dimethylaniline, can be added 
to the reaction medium in order to take up the hydrogen halide formed. 
In a preferred embodiment, the carbon tetrahalide is added, preferably in 
excess, to a starting material II in one of the inert aprotic solvents 
mentioned, such as methylene chloride, at room temperature, that is to say 
about 20.degree.-25.degree. C, and triphenylphosphine is then added in 
amounts of about 1.2 to 2 equivalents of the starting material and the 
reaction mixture is left to stand, or is stirred, at the same temperature 
until the halogenation is complete. 
Compounds I wherein Hal denotes chlorine or bromine can be obtained, for 
example, by treating the starting material II with a corresponding 
N,N-disubstituted halogeno-iminium halide compound, especially of the 
formula 
##STR1## 
and subsequently splitting off protective groups which are present. In the 
formula III, R.sub.1 and R.sub.2 represent organic, for example aliphatic, 
radicals, above all lower alkyl and especially methyl, and R.sub.3 
represents, in particular, hydrogen but can also be an organic, for 
example aliphatic, radical, such as lower alkyl, and especially methyl, 
whilst Hal.sub.1 is chlorine or bromine. 
The above reagent is usually manufactured in situ by treating a suitable 
N,N-disubstituted amide of the formula 
##STR2## 
wherein R.sub.1, R.sub.2 and R.sub.3 have the abovementioned meanings, 
especially a corresponding N,N-disubstituted formamide and above all 
dimethylformamide, with one of the chlorinating or brominating agents 
customarily used. The latter are suitable carbonic acid halides, for 
example phosgene, or carbonyl dibromide, carboxylic acid halides, for 
example oxalyl chloride or oxalyl bromide, sulphuric acid halides, for 
example thionyl chloride or thionyl bromide, or phosphoric acid chlorides, 
for example phosphorus trichloride, phosphorus oxychloride, phosphorus 
tribromide or phosphorus oxybromide, and also phosphorus pentachloride. 
Particular chlorinating and brominating agents are phosphorus trichloride 
and phosphorus tribromide. 
The above reaction is usually carried out in the presence of a solvent or 
diluent and it is usually possible to use, as the solvent or diluent, an 
amide of the formula IV which is also suitable as a solvent, especially 
dimethylformamide, which preferably is in the anhydrous form. In addition 
to the amide, which is usually present in excess and serves as the solvent 
and is customarily dimethylformamide and also dimethylacetamide, it is 
also possible correspondingly to use ether-like solvents, for example 
tetrahydrofurane or dioxane, halogenated hydrocarbons, for example 
methylene chloride, or sulphoxides, for example dimethylsulphoxide. 
The abovementioned chlorinating and brominating agents are customarily used 
in amounts which correspond to two equivalents of the 3-hydroxy-3-cephem 
starting material. The reaction can, for example, be so carried out that 
the chlorinating or brominating agent is added to a solution of the 
3-hydroxy-3-cephem starting material is an amide of the formula IV, 
especially in dimethylformamide. During the addition, this solution is 
kept at a temperature of about 0.degree. C to about 15.degree. C and 
thereafter the reaction mixture is left to stand for several hours at room 
temperature. Initially, the reaction is exothermic; the reaction vessel 
must therefore be so cooled that the temperature in this reaction phase 
can be kept below about 25.degree. C. The reaction mixture is then left to 
stand at about room temperature for the remainder of the reaction period 
and it is possible to follow the course of the reaction by thin layer 
chromatography. 
The chlorination or bromination can also be carried out by first mixing the 
chlorinating or brominating agent with the amide of the formula IV, 
especially dimethylformamide, by which means the halogeno-iminium halide 
of the formula III is formed, and then reacting the latter with a solution 
of the 3-hydroxy-3-cephem starting material II in the amide, especially in 
dimethylformamide, to which an additional solvent can also be added, or in 
another solvent, for example tetrahydrofurane. If necessary, the reactions 
are carried out in an inert gas atmosphere. 
The conversion of the 3-hydroxyl group into fluorine can be effected, for 
example, by treating the starting material II with a reagent of the 
formula F.sub.3 S-Am, wherein Am represents a disubstituted amino group; 
such reagents have been described, inter alia, by Markovsky et al., 
Synthesis, Volume 1973, page 787. The amino group preferably contains, as 
substituents, two monovalent, optionally substituted, preferably 
aliphatic, but also aromatic, hydrocarbon radicals or one divalent, 
optionally substituted, preferably aliphatic hydrocarbon radical. 
Monovalent aliphatic hydrocarbon radicals are, above all, lower alkyl, for 
example methyl, ethyl, n-propyl, isopropyl or straight-chain or branched 
butyl, whilst corresponding aromatic hydrocarbons are preferably 
optionally substituted phenyl, for example phenyl substituted by lower 
alkyl, such as methyl, lower alkoxy, such as methoxy, and/or halogen, for 
example chlorine. In a divalent aliphatic hydrocarbon radical, carbon 
atoms can optionally be replaced by hetero-atoms, such as an oxygen or an 
optionally substituted nitrogen atom; such divalent radicals are lower 
alkylene, for example 1,4-butylene or 1,5-pentylene, oxa-lower alkylene, 
for example 3-oxa-1,5-pentylene, or aza-lower alkylene which is optionally 
N-substituted by lower alkyl, for example 3-methyl-3-aza-1,5-pentylene. 
The group Am therefore above all represents di-lower alkylamino, for 
example dimethylamino, diethylamino, ethyl-methyl-amino, 
methyl-propyl-amino, di-n-propylamino or diisopropylamino, lower 
alkyl-phenyl-amino, for example methyl-phenyl-amino or ethyl-phenyl-amino, 
lower alkyleneamino, for example pyrrolidino or piperidino, oxa-lower 
alkyleneamino, for example morpholino, or amino which is optionally 
substituted by aza-lower alkyl, for example 4-methyl-piperazino. 
The above reaction is preferably carried out in the presence of a suitable 
inert solvent and the solvents used are, for example, optionally 
substituted carbocyclic hydrocarbons, for example alicyclic hydrocarbons, 
such as cyclopentane, cyclohexane, cycloheptane or decahydronaphthalene, 
or aromatic carbocyclic hydrocarbons, such as benzene, toluene or xylenes, 
which can also be halogenated on the nucleus, such as chlorobenzene, 
dichlorobenzenes or bromobenzene, and especially saturated aliphatic 
hydrocarbons, such as pentanes, hexanes, heptanes or octanes, or 
corresponding halogenated, and especially chlorinated, hydrocarbons, such 
as chloroform, 1,1- or 1,2-dichloroethane, 1,1-, 1,2- or 
1,3-dichloropropane and, above all, methylene chloride. Further solvents 
which can also be used are aliphatic, and especially cyclic, ethers, such 
as diethyl ether, diisopropyl ether, ethylene glycol dimethyl ether, 
tetrahydrofurane and, above all, dioxane, and also nitrogen-containing 
aromatic heterocyclic compounds, such as pyridine and its homologues or 
quinoline. Optionally, an excess of the fluorinating agent can be used as 
the solvent and/or several of the solvents mentioned can be combined with 
one another. 
The reaction is, if necessary, carried out with cooling or warming, for 
example in a temperature range from about -20.degree. C to about 
80.degree. C, preferably from about 0.degree. C to about 30.degree. C, 
and/or under an inert gas atmosphere. 
A fluorine atom can also be introduced when the starting material used is a 
starting material II in which the hydroxyl group is in the form of an 
organic sulphonyloxy group, that is to say when a 
7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-sulphonyloxy-3-c 
ephem-4-carboxylic acid (IIa), wherein the carboxyl group is in the 
protected form and the amino group is preferably in the protected form and 
the sulphonyloxy group represents an organic sulphonyloxy group, is 
reacted with an organic fluoride in the presence of a crown ether and, if 
necessary and desired, the additional process steps are carried out. 
An organic sulphonyloxy group is, above all, lower alkylsulphonyloxy, 
especially methylsulphonyloxy, but can also by arylsulphonyloxy wherein 
aryl is preferably phenyl which is optionally substituted, for example by 
lower alkyl, such as methyl, halogen, for example bromine, or nitro, for 
example 4-methyl-phenylsulphonyloxy. 
An inorganic fluoride is, above all, a metal fluoride and, in particular, 
an alkali metal fluoride, for example sodium fluoride, or a heavy metal 
fluoride, for example silver fluoride, is used. 
The crown ethers which are used together with the inorganic fluoride are 
optionally substituted 18-coronene 6-ethers, such as 
dicyclohexyl-18-crown-6-ether. 
The reaction is carried out in the presence of an inert solvent, especially 
a nitrile, for example acetonitrile or propionitrile, or a nitro-lower 
alkane, for example nitromethane or nitroethane, under essentially 
anhydrous conditions and, if necessary, with cooling, for example in a 
temperature range of from about -20.degree. C to about 25.degree. C, 
preferably at about room temperature, and optionally in an inert gas 
atmosphere. 
The 3-sulphonyloxy-3-cephem starting material (IIa) can also be formed in 
situ since any 
7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-sulphonyloxy-2-c 
ephem-4.xi.-carboxylic acid (IIb) which may be present together with the 
starting material IIa and wherein the carboxyl group is in a protected 
form and the amino group is preferably in a protected form and the 
sulphonyloxy group represents an organic sulphonyloxy group is converted, 
under the reaction conditions, into the corresponding 
3-sulphonyloxy-3-cephem starting material IIa and enters as such into the 
reaction with the fluorinating agent. 
The starting material II is known and can be manufactured, for example, by 
acylating the amino group in a 
7.beta.-amino-3-hydroxy-3-cephem-4-carboxylic acid (V), wherein the 
carboxyl group is in the protected form and especially in an esterified 
form, with a D-2-amino-2-(1,4-cyclohexadienyl)-acetyl radical. The 
acylation can be carried out, for example, by the method described below 
and, below, the amino group in the acylating agent is preferably in a 
protected form. The hydroxyl group in the enol grouping can be converted 
into the sulphonyloxy group, for example by treatment with an organic 
sulphonic acid halide, for example a sulphonic acid chloride, in the 
presence of a tertiary amine, such as triethylamine, or of 
dimethylformamide and propylene oxide. 
The new compounds can also be obtained when the amino group in a 
7.beta.-amino-3-Hal-3-cephem-4-carboxylic acid (VI), wherein the carboxyl 
group is preferably in a protected form and the amino group is optionally 
in a reactive, protected form, is acylated with a 
D-2-amino-2-(1,4-cyclohexadienyl)-acetyl radical, wherein the amino group 
is optionally in a protected form, and any protective groups which are 
present are subsequently split off. 
In the starting material VI, the carboxyl group can preferably be in an 
esterified form, for example as described above. The carboxylic acid 
starting material VI can, however, also be used in the form of a salt, for 
example in the form of an ammonium salt, such as a salt with 
triethylamine, or in the form of a compound which has a carboxyl group 
protected by reaction with a suitable organic phosphorus halide compound, 
such as with a lower alkyl- or lower alkoxy-phosphorus dihalide, such as 
methylphosphorus dichloride, ethylphosphorus dibromide or 
methoxyphosphorus dichloride. An amino group in a reactive protected form 
is, for example, an amino group protected by a silyl radical, such as one 
of the corresponding radicals mentioned above. 
The acylation of the free or reactive protected amino group in the starting 
material VI is carried out in a manner which is in itself known. Acylating 
agents which can be used are D-2-amino-2-(1,4-cyclohexadienyl)-acetic acid 
(VII) or a reactive derivative thereof and the amino group can usually be 
in a protected form, inter alia also in a protonised form. 
If the free acid (VII) with a protected amino group is employed for the 
acylation, suitable condensing agents, such as carbodiimides, for example 
N,N'-diethyl-, N,N'-dipropyl-, N,N'-diisopropyl-, N,N'-dicyclohexyl- or 
N-ethyl-N'-3-dimethylaminopropyl-carbodiimide, suitable carbonyl 
compounds, for example carbonyldiimidazole, or isoxazolinium salts, for 
example N-ethyl-5-phenyl-isoxazolinium 3'-sulphonate and 
N-tert.-butyl-5-methyl-isoxazolinium perchlorate, or a suitable acylamino 
compound, for example 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline, are 
customarily used. The condensation reaction is preferably carried out in 
an anhydrous reaction medium, for example in methylene chloride, 
dimethylformamide or acetonitrile. 
A functional derivative of the said acid VII, which usually has a protected 
amino group, is above all an anhydride thereof, including, and preferably, 
a mixed anhydride. Mixed anhydrides are, for example, those with inorganic 
acids, especially with hydrogen halide acids, that is to say the 
corresponding acid halides, for example the acid chloride or acid bromide, 
and also the anhydride with hydrazoic acid, that is to say the 
corresponding acid azide, with a phosphorus-containing acid, for example 
phosphoric acid or phosphorous acid, with a sulphur-containing acid, for 
example sulphuric acid, or with hydrocyanic acid. Further mixed anhydrides 
are, for example, those with organic acids, such as organic carboxylic 
acids, such as with lower alkanecarboxylic acids which are optionally 
substituted, for example by halogen, such as fluorine or chlorine, for 
example pivalic acid or trichloroacetic acid, or with half-esters, 
especially lower alkyl half-esters, of carbonic acid, such as the ethyl or 
isobutyl half-ester of carbonic acid, or with organic, and especially 
aliphatic or aromatic, sulphonic acids, for example p-toluene-sulphonic 
acid. 
Further acid derivatives which are suitable for reaction with the free 
amino group are activated esters of the said acid VII, which usually has a 
protected amino group, such as esters with vinylogous alcohols (that is to 
say enols), such as vinylogous lower alkanols, or aryl esters, such as 
phenyl esters which are preferably substituted, for example by nitro or 
halogen, such as chlorine, for example pentachlorophenyl, 4-nitrophenyl or 
2,4-dinitrophenyl esters, heteroaromatic esters, such as benztriazole 
esters, or diacyliminoesters, such as succinylimino- or 
phthalylimino-esters. 
The acylation with an acid derivative, such as an anhydride and especially 
with an acid halide, can be carried out in the presence of an acid-binding 
agent, for example an organic base, such as an organic amine, for example 
a tertiary amine, such as a tri-lower alkylamine, for example 
triethyl-amine, a N,N-di-lower alkyl-aniline, for example 
N,N-dimethyl-aniline, or a base of the pyridine type, for example 
pyridine, an inorganic base, for example an alkali metal hydroxide, 
carbonate or bicarbonate or an alkaline earth metal hydroxide, carbonate 
or bicarbonate, for example sodium hydroxide, carbonate or bicarbonate, 
potassium hydroxide, carbonate or bicarbonate or calcium hydroxide, 
carbonate or bicarbonate, or an oxirane, for example a lower 1,2-alkylene 
oxide, such as ethylene oxide or propylene oxide. 
The above acylation can be carried out in an aqueous, or preferably 
non-aqueous, solvent or solvent mixture, for example in a carboxylic acid 
amide, such as a N,N-di-lower alkylamide, for example dimethylformamide, a 
halogenated hydrocarbon, for example methylene chloride, carbon 
tetrachloride or chlorobenzene, a ketone, for example acetone, an ester, 
for example ethyl acetate, or a nitrile, for example acetonitrile, or 
mixtures thereof and, if necessary, at reduced or elevated temperature and 
optionally in an inert gas atmosphere, for example a nitrogen atmosphere. 
The starting materials (VI) are known and can be manufactured, for example, 
by splitting the acylamino grouping in 
7.beta.-acylamino-3-Hal-3-cephem-4-carboxylic acid compounds, wherein aryl 
denotes a radical which differs from the 
D-2-amino-2-(1,4-cyclohexadienyl)-acetyl radical, for example phenylacetyl 
or phenoxyacetyl, in a manner which is in itself known, for example by 
treatment with phosphorus pentachloride in the presence of pyridine, 
followed by methanol and then, optionally, water. 
The protected carboxyl and/or amino groups in the compounds obtainable 
according to the invention are optionally liberated together in a manner 
which is in itself known, such as by means of solvolysis, including 
hydrolysis, alcoholysis or acidolysis, or by means of reduction, including 
hydrogenolysis or chemical reduction. 
Thus, for example, a tert.-lower alkoxycarbonyl, polycycloalkoxycarbonyl or 
diphenylmethoxycarbonyl group can be converted into a free carboxyl group 
by treatment with a suitable acid agent, such as formic acid or 
trifluoroacetic acid, optionally with the addition of a nucleophilic 
compound, such as phenol or anisole. An optionally substituted 
benzyloxycarbonyl group can be liberated, for example, by means of 
hydrogenolysis by treatment with hydrogen in the presence of a 
hydrogenation catalyst, such as a palladium catalyst. Furthermore, certain 
substituted benzyloxycarbonyl groups, such as 4-nitrobenzyloxycarbonyl, 
can also be converted into a free carboxyl group by means of chemical 
reduction, for example by treatment with a chemical reducing agent, such 
as a metal, for example zinc, or a reducing metal salt, such as a 
chromium-II salt, for example chromium-II chloride, usually in the 
presence of a hydrogen donor which, together with the metal, is able to 
produce nascent hydrogen, such as an acid, above all acetic acid and also 
formic acid, or of an alcohol, in which case water is preferably added. It 
is also possible, in the same way, to convert a 2-halogeno-lower 
alkoxycarbonyl group (optionally after converting a 2-bromo-lower 
alkoxycarbonyl group into a 2-iodo-lower alkoxycarbonyl group) or an 
acylmethoxycarbonyl group into a free carboxyl group and an 
aroylmethoxycarbonyl group can also be converted by treatment with a 
nucleophilic, preferably salt-forming, reagent, such as sodium 
thiophenolate or sodium iodide. A carboxyl group which, for example, is 
protected by silylation can be liberated in the customary manner, for 
example by treatment with water or an alcohol. Analogously, a carboxyl 
group protected by reaction with an organic phosphorus halide compound can 
also be liberated by hydrolysis or alcoholysis. 
A protected amino group is liberated in a manner which is in itself known 
and, depending on the nature of the protective group, in diverse ways, for 
example by means of solvolysis or reduction. A 2-halogeno-lower 
alkoxycarbonylamino group (optionally after converting a 2-bromo-lower 
alkoxycarbonyl group into a 2-iodo-lower alkoxycarbonyl group), an 
acylmethoxycarbonylamino group or, for example, a 
4-nitrobenzyloxycarbonylamino group can, for example, be liberated by 
treatment with a suitable chemical reducing agent, such as zinc in the 
presence of aqueous acetic acid, or under basic conditions, for example in 
the presence of alkali metal hydroxides or secondary or tertiary amines 
with 1 to 6 equivalents of an alkali metal dithionite, for example sodium 
dithionite, a diphenylmethoxycarbonylamino, tert.-butyl-lower 
alkoxycarbonylamino or polycycloalkoxycarbonylamino group can, for 
example, be liberated by treatment with formic acid or trifluoroacetic 
acid, an optionally substituted benzyloxycarbonylamino group can, for 
example, be liberated by means of hydrogenolysis by treatment with 
hydrogen in the presence of a hydrogenation catalyst, such as a palladium 
catalyst, an arylthioamino group or aryl-lower alkylthioamino group can, 
for example, be liberated by treatment with a nucleophilic reagent, such 
as sulphurous acid, an arylsulphonylamino group can, for example, be 
liberated by means of electrolytic reduction, a 1-acyl-2-lower 
alkylideneamino group or a triarylmethyl group can, for example, be 
liberated by treatment with an aqueous mineral acid and an amino group 
protected by an organic silyl group can, for example, be liberated by 
means of hydrolysis or alcoholysis. 
Salts of the new compounds can be manufactured in a manner which is in 
itself known. Thus, salts can be formed, for example, by treatment with 
metal compounds, such as alkali metal salts of suitable carboxylic acids, 
for example the sodium salt of .alpha.-ethyl-caproic acid, or with ammonia 
or a suitable organic amine and, preferably, stoichiometric amounts, or 
only a small excess, of the salt-forming agent are used. Acid addition 
salts are obtained in the customary manner, for example by treatment with 
an acid or a suitable anion exchange reagent. Inner salts can be formed, 
for example, by neutralisation of, for example, salts, such as acid 
addition salts, to the isoelectric point, for example with weak bases, or 
by treatment with liquid ion exchangers. 
Salts can be converted into the free compounds in the customary manner, 
metal salts and ammonium salts being converted, for example by treatment 
with suitable acids and acid addition salts being converted, for example, 
by treatment with a suitable basic agent. 
The process also includes those embodiments according to which compounds 
obtained as intermediate products are used as starting materials and the 
remaining process steps are carried out with these, or the process is 
discontinued at any stage; furthermore, starting materials can be used in 
the form of derivatives or can be formed during the reaction. 
Preferably, the starting materials used and the reaction conditions chosen 
are such that the compounds mentioned initially as being particularly 
preferred are obtained. 
The new compounds of the present invention can, for example, be used to 
manufacture pharmaceutical formulations which contain an effective amount 
of the active substance together with, or mixed with inorganic or organic, 
solid or liquid excipients which can be used pharmaceutically and which 
are suitable for enteral or parenteral administration. Thus, tablets or 
gelatine capsules which contain the active compound together with 
diluents, for example lactose, dextrose, sucrose, mannitol, sorbitol, 
cellulose and/or glycine, and lubricants, for example silica, talc, 
stearic acid or salts thereof, such as magnesium stearate or calcium 
stearate, and/or polyethylene glycol are used; tablets also contain 
binders, for example magnesium aluminium silicate, starches, such as maize 
starch, wheat starch, rice starch or arrowroot, gelatine, tragacanth, 
methylcellulose, sodium carboxymethylcellulose and/or 
polyvinylpyrrolidone, and, if desired, disintegrating agents, for example 
starches, agar or alginic acid or a salt thereof, such as sodium alginate, 
and/or effervescent mixtures, or adsorbents, dyestuffs, flavourings and 
sweeteners. Furthermore, the new pharmacologically active compounds can be 
used in the form of injectable formulations, for example formulations 
which can be administered intravenously, or of infusion solutions. Such 
solutions are preferably isotonic aqueous solutions or suspensions and 
these can be prepared before use, for example from lyophilised 
preparations which contain the active compound on its own or together with 
an excipient, for example mannitol. The pharmaceutical formulations can be 
sterilised and/or contain auxiliaries, for example preservatives, 
stabilisers, wetting agents and/or emulsifiers, solubilising agents, salts 
for regulating the osmotic pressure and/or buffers. The present 
pharmaceutical formulations which, if desired, can contain further 
pharmacologically valuable substances, are manufactured in a manner which 
is in itself known, for example by means of conventional mixing, 
granulating, dragee-making, dissolving or lyophilising processes and 
contain from about 0.1% to 100%, and especially from about 1% to about 
50%, of the active compound and lyophilisates contain up to 100% of the 
active compound. The individual dose for a warm-blooded animal weighing 
about 70 kg is between 0.1 and 0.75 g and the daily dose is between 0.2 
and 1.0 g. 
In the context of the present description, the organic radicals designated 
as "lower" contain, where they are not expressly defined, up to 7 and 
preferably up to 4 carbon atoms.

The examples which follow serve to illustrate the invention; the 
temperatures are given in degrees centigrade. 
EXAMPLE 1 
a. 100 ml of dimethylformamide are cooled to 0.degree. C and 4.9 ml (66.6 
mmols) of thionyl chloride are added slowly. 9.75 g (16.7 mmoles) of 
benzhydryl 
7.beta.-[D-2-(tert.-butoxy-carbonylamino)-2-(1,4-cyclohexadienyl)-acetylam 
ino[-3-hydroxy-3-cephem-4-carboxylate are added, in portions, to this 
solution under N.sub.2 and the temperature is brought slowly to 25.degree. 
C. After stirring for 4 hours at room temperature and under N.sub.2, a 
further 2.5 ml of thionyl chloride are added. After a total reaction 
period of 41/2 hours, the crude product is poured onto about 500 ml of ice 
and 400 ml of ethyl acetate, the mixture is diluted with 500 ml of water 
and the organic phase is separated off. The organic phase is washed with 
five times 700 ml of water and a little saturated NaCl solution. The 
aqueous phases are extracted twice more with ethyl acetate and the 
combined ethyl acetate solutions are dried over sodium sulphate and 
concentrated. The crude product is subjected to column chromatography (20 
times the amount of silica gel, system: toluene/ethyl acetate, 4:1). The 
corresponding fractions containing benzhydryl 
7.beta.-[D-2-(tert.butoxycarbonylamino)-2-(1,4-cyclohexadienyl)-acetylamin 
o]-3-chloro-3-cephem-4-carboxylate (Rf = 0.4, silica gel, toluene/ethyl 
acetate, 3:1) are combined and employed in the next reaction step. 
For further purification, the crude product can be made to crystallise from 
methylene chloride/diethyl ether and crystals which have a melting point 
of 158.degree.-160.degree. C are obtained; UV spectrum (ethanol): 
.lambda..sub.max = 268 nm (.epsilon. = 7,400); IR spectrum (CH.sub.2 
Cl.sub.2): bands at 2.95, 5.57, 5.8, 5.9 and 6.70 .mu.. 
ai. It is also possible to employ 35 mmols of phosphorus trichloride in 
place of 66.6 mmols of thionyl chloride. 
b. 1.57 g (2.6 mmols) of benzhydryl 
7.beta.-[D-2-(tert.-butoxycarbonylamino)-2-(1,4-cyclohexadienyl)-acetylami 
no]-3-chloro-3-cephem-4-carboxylate are suspended in 2 ml of anisole at 
0.degree. C and 10 ml of trifluoroacetic acid are added. After stirring 
for 15 minutes at 0.degree. C, the solution is diluted with toluene and 
then concentrated to about 5 g in vacuo. It is then partitioned between 
water and ethyl acetate and the organic phase is washed once with water. 
The pH of the combined aqueous phases is adjusted to a pH of 4.8 with 
triethylamine, whereupon the inner salt of 
7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-chloro-3-cephem- 
4-carboxylic acid crystallises out as the dihydrate; melting point 
160.degree. C (decomposition); UV spectrum (0.1 N HCl): .lambda..sub.max = 
267 nm (.epsilon. = 7,700); IR spectrum (Nujol): bands at 2.98, 5.6, 5.76, 
6.2 and 6.64 .mu.. 
The starting material can be obtained as follows: 
c. 2.17 g (4.16 mmols) of benzhydryl 
7.beta.-amino-3-hydroxy-3-cephem-4-carboxylate are dissolved in 32 ml of 
methylene chloride (anhydrous) and the solution is then stirred with 1.53 
ml (6.3 mmols) of bis-trimethylsilyl-acetamide at room temperature for 40 
minutes under N.sub.2 (solution I). A further solution (II) of 1.77 g (7 
mmols) of D-2-tert.-butoxy-carbonylamino-2-(1,4-cyclohexadienyl)-acetic 
acid in 75 ml of methylene chloride is stirred with 0.78 ml (7.0 mmols) of 
N-methyl-morpholine and 0.91 ml (7.0 mmols) of isobutyl chloroformate for 
30 minutes at 0.degree. C under nitrogen. Solution I is cooled to 
-10.degree. C and solution II is added slowly. The reaction mixture is 
stirred for 30 minutes at -10.degree. C and for 30 minutes at 0.degree. C, 
100 ml of water are added and the phases are separated. The organic 
solution is washed with a saturated aqueous solution of sodium chloride, 
dried over sodium sulphate and concentrated. The residue is purified by 
means of preparative layer chromatography (silica gel, system: 
toluene/ethyl acetate, 3:1, Rf about 0.15). Benzhydryl 
7.beta.-[D-2(tert.-butoxycarbonylamino)-2-(1,4-cyclohexadienyl)-acetylamin 
o[-3-hydroxy-3-cephem-4-carboxylate, which according to thin layer 
chromatography is a single compound, is obtained as an amorphous product. 
EXAMPLE 2 
420 mg (1.6 mmols) of triphenylphosphine are added, at room temperature and 
under a nitrogen atmosphere, to a solution of 617 mg (1 mmol) of 
benzhydryl 7.beta.-[D-2-(tert.-butoxy-carbonylamino)-2-(1,4-cyclohexadieny 
l)-acetylamino]-3-hydroxy-3-cephem-4-carboxylate in a mixture of 1 ml of 
carbon tetrachloride and 20 ml of methylene chloride. The reaction mixture 
is left to stand at room temperature for 24 hours and then washed with 
water and a saturated aqueous solution of sodium chloride. The organic 
phase is dried over sodium sulphate and evaporated in vacuo. The residue 
is chromatographed on silica gel using toluene/ethyl acetate, 3:1. The 
fractions containing benzhydryl 
7.beta.-[D-2-(tert.-butoxy-carbonylamino)-2-(1,4-cyclohexadienyl)-acetylam 
ino[-3-chloro-3-cephem-4-carboxylate are combined and crystallised from 
methylene chloride/diethyl ether. The resulting crystals having a melting 
point of 158.degree.-160.degree. C. 
EXAMPLE 3 
460 mg (1.3 mmols) of dichloro-triphenyl-phosphorane and 0.08 ml (1 mmol) 
of pyridine are added, at room temperature and under a nitrogen 
atmosphere, to a solution of 617 mg (1 mmol) of benzhydryl 
7.beta.-[D-2-(tert.-butoxycarbonylamino)-2-(1,4-cyclohexadienyl)-acetylami 
no]-3-hydroxy-3-cephem-4-carboxylate in 10 ml of methylene chloride. The 
reaction mixture is left to stand at room temperature for 24 hours and 
then washed with water and a saturated aqueous solution of sodium 
bicarbonate. The organic phase is dried over sodium sulphate and 
evaporated in vacuo. The residue is chromatographed on silica gel using 
toluene/ethyl acetate, 3:1. The fractions containing benzhydryl 
7.beta.-[D-2-(tert.-butoxy-carbonylamino)-2-(1,4-cyclohexadienyl)-acetylam 
ino]-3-chloro-3cephem-4- carboxylate are combined and crystallised from 
methylene chloride/diethyl ether. The resulting crystals have a melting 
point of 158.degree.-160.degree. C. 
EXAMPLE 4 
A solution, which has been cooled to 0.degree. C, of 0.253 g (1 mmol) of 
D-.alpha.-tert.-butoxycarbonylamino-.alpha.-(1,4-cyclohexadienyl)-acetic 
acid in 75 ml of methylene chloride is stirred with 0.097 ml of 
N-methyl-morpholine and 0.129 ml of isobutyl chloroformate under a 
nitrogen atmosphere for 30 minutes, the mixture is then cooled to 
-10.degree. and 0.33 g of benzhydryl 
7.beta.-amino-3-chloro-3-cephem-4-carboxylate and 0.085 ml of 
N-methyl-morpholine are added successively. The reaction mixture is 
stirred for 30 minutes at -10.degree. C and for 30 minutes at 0.degree. C, 
30 ml of water are added and the pH value is adjusted to 7.9 by adding a 
40% strength aqueous solution of dipotassium hydrogen phosphate. The 
phases are separated, the aqueous solution is extracted with methylene 
chloride and the combined organic solutions are washed with a saturated 
solution of sodium chloride, dried over sodium sulphate and evaporated 
under reduced pressure. The residue is purified by means of preparative 
layer chromatography (silica gel, system: toluene/ethyl acetate, 3:1, Rf 
about 0.4). Benzhydryl 
7.beta.-[D-2-(tert.-butoxycarbonylamino)-2-(1,4-cyclohexadienyl)-acetylami 
no]-3-chloro-3-cephem-4-carboxylate, which according to thin layer 
chromatography is a single compound, is obtained as an amorphous product 
which can be further used analogously to Example 1. 
EXAMPLE 5 
0.4 ml of bis-(trimethylsilyl)-acetamide is added to a suspension of 323 mg 
(1.38 mmols) of 7.beta.-amino-3-chloro-3-cephem-4-carboxylic acid in 10 ml 
of methylene chloride and the mixture is stirred for 1 hour at room 
temperature. The solution, which is then clear, is cooled to -10.degree. C 
and 0.315 g (1.52 mmols) of 2-(1,4-cyclohexadienyl)-glycyl 
chloride-hydrochloride is added. The reaction mixture is stirred at the 
same temperature for 1 hour and 20 ml of water are then added. The aqueous 
phase is washed with about 20 ml of methylene chloride and the pH value is 
raised to 5.5 by adding 1 N sodium hydroxide solution. After concentrating 
the aqueous phase, the inner salt of 
7.beta.-[D-2-amino-(1,4-cyclohexadienyl)-acetylamino]-3-chloro-3-cephem-4- 
carboxylic acid crystallises out as the dihydrate. The melting point, the 
UV spectrum and the IR spectrum correspond to the values given in Example 
1b. 
EXAMPLE 6 
7.beta.-[D-2-Amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-fluoro-3-cephem-4 
-carboxylic acid and 
7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-bromo-3-cephem-4 
-carboxylic acid, or their salts, especially their inner salts, can be 
manufactured in the manner described and illustrated above. 
EXAMPLE 7 
Dry ampoules or phials containing 0.5 g of the inner salt of 
7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-chloro-3-cephem- 
4-carboxylic acid are manufactured as follows: 
______________________________________ 
Composition (for 1 ampoule or phial) 
______________________________________ 
inner salt of 7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)- 
acetylamino]-3-chloro-3-cephem-4-carboxylic acid 
0.5 g 
mannitol 0.05 g 
______________________________________ 
A sterile aqueous solution of the inner salt of 
7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-chloro-3-cephem- 
4-carboxylic acid and mannitol is subjected to freeze-drying under aseptic 
conditions in 5 ml ampoules or 5 ml phials and the ampoules or phials are 
sealed and tested. 
EXAMPLE 8 
Capsules containing 0.25 g of the inner salt of 
7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-chloro-3-cephem- 
4-carboxylic acid are manufactured as follows: 
______________________________________ 
Composition (for 4,000 capsules) 
______________________________________ 
inner salt of 7.beta.-[D-2-amino-2-(1,4-cyclohexa- 
dienyl)- 
acetylamino]-3-chloro-3-cephen-4-carboxylic acid 
250,000 g 
maize starch 50,000 g 
polyvinylpyrrolidone 15,000 g 
magnesium stearate 5,000 g 
ethanol q.s. 
______________________________________ 
The inner salt of 
7.beta.-[D-2-amino-2-(1,4-cyclohexadienyl)-acetylamino]-3-chloro-3-cephem- 
4-carboxylic acid and the maize starch are mixed and the mixture is 
moistened with a solution of polyvinylpyrrolidone in 50 g of ethanol. The 
moist mass is pressed through a sieve which has a mesh width of 3 mm and 
dried at 45.degree.. The dry granules are forced through a sieve which has 
a mesh width of 1 mm and mixed with 5 g of magnesium stearate. The mixture 
is filled, in portions of 0.320 g, into size 0 push-fit capsules.