CA1055924A - Methoxy-heterocyclic compounds containing aminomethyl groups - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE
6.beta.-Aminomethylaryl-acetylemino-6.alpha.-methoxy-2,2-dimethyl-penam-3-carboxylic acid compounds and 7.beta.-aminomethylaryl-acetylamino-7.alpha.-methoxy-3-cephem-4-carboxylic acid compounds, wherein the aminomethyl-substituted aryl radical represents a corresponding thienyl, furyl or phenyl radical, and wherein the 3-position in 3-cephem compounds is unsubstitued or substituted by a modified hydroxyl group or an optionally substituted methyl group, exhibit antibiotic actions against Gram-negative or Gram-positve micro-organisms.

Description

The present inVention relates to a process for the manufacture of 7~-acylamino-7~-methoxy-3-cephem-4-carboxylic acid compounds of the formula H N -CH ~ ~ N ~IC-R1 (I), o=C- R
wherein X represents sulphur or oxygen~ or represents ethenylene of the formula -CR=CH-~ and wherein Rl represents lower alkoxy or a radical of the formula -CH2 -R27 wherein R2 denotes a hydroxyl or mercapto group etherified by lower alkyl~
or heterocyclylthio~ wherein heterocyclyl represents a monocyclic five-membered heterocyclic radical of aromatic character whiCh is bonded to the thiosulphur atom via a ring carbon atom and which contains 2 or 3 ring nitrogen atoms and optionally addition-ally a ring oxygen atom~ ring sulphur atom or ring nitrogen atom, and such a radical can optionally be substituted by lower alkyl~
or wherein R2 denotes lower alkanoyloxy~ optionally N-lower alkylated or N-halogeno-lower alkylated carbamoyloxy and R represents hydroxyl or an etherified hydroxyl group which together with the carbonyl grouping -C(=O~- forms an esterified carboxyl group which can be split under physiological conditions, and of pharmaceutically acceptable salts thereof.

-2-li~sss~
The group ~ is above all sulphur, but it can also be oxygen, or ethenylene of the formula -CH=CH-. The amino-methyl-substituted radical ther~re represe~s a~nome'-h~l~en~l for example 4- or 5-aminomethyl-2- or -3-thienyl~ or 3-aminomethyl-2-thienyl or 2-aminomethyl-3-thienyl, or corres-ponding aminomethyl-furyl, for example 4- or 5-aminometh~Jl-2-furyl, or aminomethyl-phenyl, for e~am~le 2- or 4-amino-methylphenyl.
As a lower alkoxy group~Rl represen~s a group with up to 7, in particular with up to 4, carbon atoms, above all methoxy, and also etho~y, n-propoxy or isopropoxy, as well as straight-chain or branched butoxy, pentoxy, hexyloxy or heptyloxy.
As a hydroxyl or mercapto group etherified by lower alkyl, R2 represents lower alkoxy as defined above for a group Rl or lower alkylthio, with up to 7, in particular with up to 4, carbon atoms, above all methylthio, as well as ethylthio, n-propylthio or isopropylthio, and also straight-chain or branched butylthio, pentylthio, hexylthio or heptylthio.
In a heterocyclylthio group R2 substituted by lower alkyl, lower alkyl is especially methyl, as well as ethyl, n-propyl, isopropyl or straight-chain or branched butyl, pentyl or hexyl.
Such five-membered heterocyclic radicals are, for example, diazacyclic, triazacyclic, tetrazacyclic, thiadiaza-cyclic, thiatriazacyclic or oxadiazacyclic radicals, which are optionally substituted by lower alkyl, for example methyl.

~)S55~;~4 Preferred heterocyclic etherified mercapto groups R2 are, inter alia, imidazolylthio, for example 2-imidazolylthio, optionally lower alkyl-substituted triazolylthio, for example l-methyl-lH-1,2,3~triazol-4-ylthio, lH-1,2,4-tria-zol-3-ylthio, 5-methyl-lH-1,2,4-triazol-3-ylthio, or 4,5-dimethyl-4H-1,2,4-triazol-3-ylthio, optionally lower alkyl-substituted tetrazolylthio, for ex-ample lH-tetrazol-5-ylthio, or l-methyl-lH tetrazol-5-ylthio, optionally lower alkyl-substituted thiadiazolylthio, for example 1,2,3-thiadiazol-4-ylthio, 1,2,3-thiadiazol-5-ylthio, 1,3,4-thiadiazol-2-ylthio, 2-methyl-1,3,4-thiadiazol-5-ylthio, 1,2,4-thiadiazol-5-ylthio or 1,2,5-thiadiazol-3-ylthio, thiatriazol-10 ylthio, for example 1,2,3,4-thiatriazolyl-5-ylthio, optionally lower alkyl-substituted optionally lower alkyl-substituted oxadiazolylthio, for example 1,2,4-oxadiazol-5-ylthio, or 2-methyl-1,3,4-oxadiazol-5-ylthio.
Hydroxyl groups R2 esterified with lower alkanoyloxy, are especially acetoxy, and also formyloxy, propionyloxy, valeryloxy, hexanoyloxy, heptanoyl-oxy or pivalyloxy.
An esterified hydroxyl group R2 is furthermore a hydroxyl group esterified by an optionally N-substituted half-amide of carbonic acid. N-substituents are lower alkyl optionally containing halogen, for example chlor-ine, for example methyl, ethyl or 2-chloroethyl. Hydroxyl groups R2 esteri-fied in this way are, for example, carbamoyloxy, N-methyl-carbamoyloxy, N-ethylcarbamoyloxy or N-(2-chloroethyl)-carbamoyloxy.
In an esterified carboxyl group of the formula -C(=0)-R which can be split under physiological conditions, R is above all an acyloxymethoxy group, wherein acyl denotes, for example, the radical of an organic carboxylic acid, above all of an optionally substituted lower alkanecarboxylic acid, or wherein acyloxymethyl forms the radical of a lactone. Such groups R2 are lower alkanoyloxymethoxy, for example . 4 1~5~

acetoxymethoxy or pivaloyloxymethoxy 9 amino-lower alkanoyloxy-methoxy, especially a-amino-lower alkanoyloxymethoxy, for example glycyloxymethoxy, L-valyloxymethoxy or L-leucyloxy-methoxy, and also phthalidyloxy, for example 2-phthalidyloxy, or indanyloxy, for example 5-indanyloxy.
Salts are9 in particular, salts of compounds of -the formula I having a free carboxyl group -C(=O)-R, above all metal salts or ammonium salts, such as alkali metal salts and alkaline earth metal salts, for example sodium, potassium, magnesium or calcium salts, as well as ammonium salts with ammonia or suitaole organic amines, the amines which can be used for salt formation being above all aliphatic, cyclo-aliphatic, cycloaliphatic-aliphatic or araliphatic primary, secondary or tertiary monoamines, diamines or polyamines, as well as heterocyclic bases, such as lower alkylamines, for example triethylamine, hydroxy-lower alkylamines, for example 2-hydroxyethylamine, bis-(2-hydroxyethyl)-amine or tris-(2-hydroxyethyl)-amine, basic aliphatic esters of carboxylic acids, for example 4-aminobenzoic acid 2-diethylaminoethyl ester, lower alkyleneamines, for example l-ethyl-piperidine, cycloalkylamines, for example dicyclohexylamine, or benzyl-amines, for example N,N'-dibenzyl-ethylenediamine, as well as bases of the pyridine type, for example pyridine, collidine or quinoline. Compounds of the formula I 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 acids or sulphonic acids, for example trifluoroacetic acid, and with aminoacids, such as , - ~, _ ,~Z _ 1t~559~4 arginine and lysine. Compounds of the formula I having a free carboxyl group can also be in the form of inner salts, that is to say in the form of zwitter-ions.
The compounds of the formula I and their pharma-ceutically usable non-toxic salts are valuable antibiotically active substances ~hich can in particular be used as anti-bacterial antibiotics. For example, they are active against micro-organisms, such as against Gram-positive bacteria, for example against Staphylococcus aureus (in vitro in minimum concentrations of about 0.001 mg/ml and, for example in mice, in doses of about 2 to about 10 mg/kg given subcutaneously), including against penicillin-resistant Staph~lococcus aureus (in v tro in minimum concentrations of about 0.001 mg/ml), and also against Bacillus subtilis (in vitro in minimum concentrations of about 0.001 mg/ml) and against Gram-negative bacteria, for example against Escherichia coli (in vi-tro in minimum concentrations of about 0.005 mg/ml and, for example, in mice, in doses of about 10 to about 100 mg/kg administered subcutaneously), including against ampicillin-resistant, carbenicillin-resistant and rifamycin-resistant Escherichia coli (in vitro in minimum concentrations of about 0.005 mg/ml), .
and also against Klebsiella pneumoniae and Salmonella t~r3himurium, including agains~ ampicillin-resistant, carbeni-cillin-resistant and rifamycin-resistant Salmonella typhimurium (in vitro in minimum concentrations of about 0.005 g/ml), Proteus vul~aris, Proteus mirabilis, including carbenicillin-resistant Proteus mirabilis, and Proteus rett~eri (in vitro in minimum concentrations of about G

--,a~--.
;

S9;~4 0.03 mg/ml~. The new compounds are distinguished by excellent stability to -lactamases, such as cephalosporinases, especially of Gram-negative bacteria, as can be demonstrated from the rates of hydrolysis in the presence of iso-lated ~lactamases from various Gram-negative germs, such as Escherichia coli, Aerobacter colacae, Proteus morganii and Pseudomonas aeruginosa. The rates of hydrolysis of the new compounds by ~lactamases are, for example, more than a hundred times lower than those of cephalothin and cephaloridin. The new compounds can therefore be used correspondingly, for example in the form of antibiotically active preparations, for the treatment of infections caused by Gram-positive or Gram-negative bacteria.
Above all, the present invention relates to those compounds of the formula I wherein X represents oxygen or, in particular, sulphur, or ethenylene of the formula -CHeCH-, and the aminomethyl-substituted radical represents amino-methyl-2-thienyl, such as 4- or 5-, as well as 3-amino-methyl-2-thienyl, as well as aminomethyl-3-thienyl, and also aminomethyl-2-furyl, such as 4- or 5- as well as 3-amino-methyl-2-furyl, as well as aminomethyl-3-furyl, and aminomethyl-phenyl, for example 2- or 4-aminomethyl-phenyl, wherein Rl denotes lower alkoxy, preferably with up to 4 carbon atoms, or the group of the formula -CH2-R2, and R2 represents lower alkanoyloxy, especially acetoxy, optionally lower alkylated or halogenoalkylated carbamoyloxy, lower alkylthio or option-ally lower alkylated heterocyclylthio, and wherein R represents hydroxyl, as well as salts, especially the non-toxic, pharmaceutically usable salts, parti-cularly the alkali metal salts or alkaline earth metal salts, as well as the inner salts, of such compounds.
Above all, the present invention relates to compounds of the formula I wherein X represents oxygen or in particular sulphur, and also ethenylene of the formula -CH=CH-, and the aminomethyl-substituted radical represents aminomethyl-2- or -3-thienyl, for example 4- or 5- as well as 3-aminomethyl-2-thienyl, and also aminomethyl-2-furyl, for example 4- or 5- aminomethyl-2-furyl, as well as aminomethyl-phenyl, for example 2- or 4-aminomethyl-phenyl, and wherein Rl denotes lower alkoxy with up to 4 carbon atoms, such as methoxy, or the group of the formula -CH2R2, and R2 denotes lower alkanoyloxy, for ex-~5~4 ample acetoxy, optionally N-lower alkylated, as well as N-halogeno-lower-alky-latedJ carbamoyloxy, for example carbamoyloxy, methylcarbamoyloxy, ethylcar-bamoyloxy or 2-chloroethyl-carbamoyloxy, lower alkylthio, for example methyl-thio, optionally lower alkylated, such as methylated heterocyclylthio, and wherein R represents hydroxyl, as well as salts, in particular pharmaceutical-ly usable non-toxic salts, especially the alkali metal salts or alkaline earth metal salts, as well as the inner salts, of such compounds.
In particular, the invention relates to 3-cephem-compounds of the formula I, wherein X above all represents sulphur, and also oxygen, and the aminomethyl-substituted radical denotes aminomethyl-2-thienyl or -2-furyl, for example 4- or preferably 5-, and also 3-aminomethyl-2-thienyl or 4- or 5-aminomethyl-2-furyl, and wherein Rl represents lower alkoxy with up to 4 car-bon atoms, especially methoxy, or the radical of the formula -CH2-R2, wherein R2 denotes acetoxy, carbamoyloxy, N-lower alkylcarbamoyloxy, for example methylcarbamoyloxy or ethylcarbamoyloxy, N-halogeno-lower alkyl-carbamoyloxy, for example 2-chloroethylcarbamoyloxy, lower alkylthio, for example methyl-thio, thiadiazolylthio which is optionally substituted by lower alkyl, for example methyl, and is bonded to the thio sulphur atom via a ring carbon atom, for example 1,3,4-thiadiazol-2-ylthio, 5-methyl-1,3,4-thiadiazol-2-ylthio or 5-methyl-1,2,4-thiadiazol-2-ylthio, or tetrazolylthio which is optionally sub-stituted by lower alkyl, for example methyl, and is bonded to the thio sulphur atom via a ring carbon atom, for example l-methyl-5-tetrazolylthio, and wherein the group R represents hydroxyl, as well as salts, in particular the non-toxic pharmaceutically usable salts, especially the alkali metal salts or alkaline earth metal salts, as well as the inner salts, of such compounds.
Above all, the invention relates to 3-cephem compounds of the formula I, wherein X above all represents sulphur and also oxygen, and the aminomethyl-substituted radical denotes aminomethyl-2-thienyl or -2-furyl, for example 4-or preferably 5- and also 3-aminomethyl-2-thienyl or -2-furyl, and wherein R
represents methoxy or the radical of the formula -CH2R2, wherein R2 denotes acetoxy, ~ -8-~ J~
carbamoyloxy, methylcarbamoyloxy, ethylcarbamoyloxy, 2-chloro-ethylcarbamoyloxy, methylthio, 5-methyl-1,3,4-thiadiazol-2-ylthio or l-methyl-5-tetrazolylthio, and wherein R represents hydrogen, as well as salts, especially the non-toxic pharma-ceutically usable salts, especially the alkali metal salts or alkaline earth metal salts, as well as the inner salts, of such compounds.
In particular, the invention relates to the compounds described in the examples as well as to their salts, especi-ally the non-toxic pharmaceutically usable salts, such as the alkali metal salts or alkaline earth metal salts, and above all their inner salts, which at the doses stated exhibit excellent antibiotic effects and are used correspondingly in the form of antibiotically active preparations.
m e new compounds of the present invention can be manufactured in a manner which is in itself known, for example by a method in which, in a compound of the formula I S
2 ~ ~clH2 (II) o ~ "C-R

Ro' ~0 wherein the amino group can optionally be substituted by a group which permits the acylation, wherein Rl has the above mentioned meaning 10555~;~4 ~nd wherein Ro has the meaning of R or represents a carboxyl protective radical which forms a protected carboxyl group with the carbonyl group of the formula -C(=O)-, or in a salt thereof, the amino group is acylated by treatment with an acid of the formula H2N_CH2 t CH2-C-OH (III) X

wherein the amino group is protected if appropriate, or with a reactive func-tional acid derivative thereof or with a salt of such a compound and, in a resulting compound, protected amino of the aminomethyl radical is converted into free amino and, if desired or required, a carboxyl group of the formula -C(=O)-Ro is converted into a carboxyl group of the formula -C(=O)-R and/or, if desired, a group Rl is converted into another group Rl and/or, if desired, a resulting salt is converted into the free compound or into another salt or a resulting free compound is converted into a salt.
Radicals which may be present, in s starting material of the formula II, as substituents of the amino group which , -10-. Y~

l~S~ 4 permit the acylation of the latter are, for example, organic silyl or stannyl groups, &nd also ylidene groups which together with the amino groups form a Schiff's base. The said org&nic silyl or stannyl groups are, for example, the same groups which are also capable of forming a protected carboxyl group -C(=O)-Ro with the carboxyl group on the penam or cephem ring. When silyla-ting or stannylating a carboxyl group in a starting material of the formula II,-the amino group can also be silylated or stannylated if &n excess of the silylating or stannylating agent is used.
The ylide~e groups mentioned are above all arylmethylene groups~ wherein aryl in particular represents a carbocyclic, above all monocyclic, aryl radical, for example represents phenyl which is optionally substituted, such as phenyl sub-stituted by ni-tro or hydroxyl; such arylmethylene groups are, for example benzylidene, 2-hydroxybenzylidene or 4-nitro-benzylidene, and also oxacycloalkylidene which is optionally substituted, for example by carboxyl, for example 3-carboxy-2-oxacyclohexylidene.
A protected carboxyl group of the fGrmula -C(=O)-Ro in a starting material of the formula II is above all an esterified carboxyl group which C&n preferably be split easily &nd wherein Ro represents an etherified hydroxyl group, or a carboxyl group present in the form of an anhydride, wherein Ro denotes an esterified, and in particular a phos-phorylated~hydroxyl group.
An etherified hydroxyl group Ro which, in the s-tarting material of the formula II, forms a preferably easily r ~
.~ .

lV~ 4 splittable esterified carboxyl group with the carbonyl grouping of the formula -C(=O)- is, for example, a lower alkoxy group which is preferably substi-tuted, above all in the a-position and also in the ~-position)and/or is branched in the a-position. Substituents of such a group are, for example, carbocyclic aryl, such as phenyl which is optionally substituted, for example by lower alkyl, such as tert.-butyl, phenyl, hydroxyl, lower alkoxy, such as methoxy, and/or nitro, furyl, such as 2-furyl, aryloxy, such as phenyloxy which is optionally substituted, for example by lower alkoxy, such as methoxy, arylcarbonyl, such as benzoyl which is optionally substituted, for example by halogen, such as bromine, nitrile or acylamino 9 such as diacylamino, for example phthalimino or succinylimino; such substituents are preferably in the a-position of the lower alkoxy group Ro and the latter can con- -tain one, two or more such radicals~depending on the nature of the substituents. Further substituents :7hich are prefer-ably present in the ~-position of -the lower alkoxy radical Ro are halogen, for example chlorine, bromine or iodine, and in such radicals an individual chlorine or bromine can easily be con~erted into iodine before liberating a carboxyl group protected in this way. Examples of the abovementioned op-tionally substituted lower alkoxy groups Ro are tert.-lower alkoxy, for example tert.-butoxy or tert.-pentoxy, a-phenyl-lower alkoxy which is optionally substituted in the phenyl radical, for example as indicated, such as benzyl-oxy, 4-hydroxy-3,5-di-tert.-butyl-benzyloxy, 2-biphenylyl-2-propoxy, 4-methoxy-benzyloxy, 4,5-dimethoxy-2-nitro-benzyloxy ., ~
,' 1~5~9'~4 or 4-ni.tro-ben~yloxy, diphenylmethoxy which is optionally substituted in the phenyl radicals, for example as indicated, in particular by lower alkoxy, for example methoxy, such as benzhydryloxy or 4,4'-dimethoxydiphenylmethoxy, as well as trityloxy, bis-phenyloxy-methoxy which is optionally sub-stituted in the phenyl radicals, for example as indicated, especially by lower alkoxy, such as bis-4 methoxyphenyloxy-methoxy, phenacyloxy which is optionally substituted, in particular by halogen, such as phenacyloxy or 4-bromo-phenacyloxy, cyanomethoxy, diacyliminomethoxy, such as phthalyliminomethoxy or succinyliminomethoxy, or 2-halogeno-lower alkoxy, such as 2?2,2-trichloroethoxy, 2-bromoethoxy or 2-iodoethoxy~
Further, an etherified hydroxyl group Ro~ which together with the carbonyl grouping of the formula -C(=O)-forms a splittable, preferably easily splittable, esterified carboxyl group can also denote a cycloalkoxy group of which the ~-position is preferably a bridgehead carbon atom. Such a cycloalkoxy group Ro is, for example, l-adamantyloxy.
Further etherified hydroxyl groups which represent the radical Ro are organic silyloxy or stalmyloxy groups wherein organic radicals, of which 1 to 3 can be present, are, in particular, optionally substituted aliphatic hydrocarbon radicals, such as lower alkyl, for example methyl, ethyl, n-propyl or tert.-butyl, or halogeno-lower alkyl, for example chloromethyl or 2-chloroethyl, as well as optionally substitu-ted cycloaliphatic, aromatic or araliphatic hydrocarbon radi-cals, such as cycloalkyl, phenyl or phenyl-lower alkyl, and . ._~

~ 0~5~

also organic-substituted functional groups, such as e-theri-fied hydroxyl groups, for example lower alkoxy, such as methoxy or ethoxy, and which can optionally contain, for example, halogen, such as chlorine, as further substi-tuents.
Such radicals Ro are, inter alia1 tri-lower alkylsilyloxy, for example trimethylsilyloxy or tert.-butyldimethylsilyloxy, lower alkoxy-lower alkyl-halogeno-silyloxy, for example chloro-methoxy-methyl-silyloxy, or tri-lower alkylstannyloxy, for example tri-n-butylstannyloxy.
The group Ro can also represent a phosphoryloxy group which contains a substituted trivalent or pentavalent phos-phorus atom, and which together with the carboxyl grouping of the formula -C(-O)- forms a pro-tected carboxyl group.
Substituents of trivalent phosphorus, which can be identical or different, are, inter alia, optionally substituted hydro-carbon radicals, such as corresponding aliphatic or arali-phatic hydrocarbon radicals, for example lower alkyl or halogeno-lower alkyl, such as methyl, ethyl or chloromethyl, or phenyl-lower alkyl, such as benzyl, etherified hydroxyl or mercapto groups, such as hydroxyl or mercapto groups etherified by optionally substituted aliphatic, aromatic or araliphatic hydrocarbon radicals, for example lower alkoxy or lower alkylthio, such as methoxy, ethoxy, methylthio or n-butylthio, phenyloxy or phenylthio which are optionally substituted, for example by lower alkyl, lower alkoxy or halogen, or phenyl-lower alkoxy or phenyl-lower alkylthio which are optionally substituted, for example by lower alkyl, lower alkoxy or halogen, for example benzyloxy or benzylthio, ll~S~ 4 halogen, for example fluorine, chlorine or bromine, and/or a bivalent hydrocarbon radical which is optionally substituted and/or interrupted by hetero-atoms, such as oxygen or sulphur, such as a corresponding aliphatic or araliphatic radical, for example lower alkylene, such as 1,4-butylene or 1,5-pentylene, or l-oxa-lower alkylene, in which the second methylene group, bonded to the phosphorus atom, can optionally also be replaced by an oxygen atom or a sulphur atom, for example by l-oxa-1,4-pentylene, 1-oxa-1,5-pentylene or 1,5-dioxa-1,5-pentylene, or two hydroxyl groups etherified by a bivalent optionally substituted hydrocarbon radical, such as a corresponding ali-phatic, aromatic or araliphatic radical, such as lower alkyl-ene or 1,2-phenylene. Substituents of pentavalent phosphorus are the same as those of trivalent phosphorus and, in additionJ
an oxo group.
In a starting material of the formula II, further free functional groups which may be present in addition to the carboxyl group of the formula -C(=O)-Ro~ such as a free hydroxyl group R2, are, if desired or required~ usually pre-sent in a protected, preferably easily splittable, form during the acylation reaction; a free hydroxyl group can, for example, be present in an easily splittable etherified or esterified form, for example in the form of a lower alkoxy group, for example a methoxy group, or of a 2-oxacycloalkoxy group, for example a 2-tetrahydropyranyloxy group, or of an acyloxy group, such as a lower alkanoyloxy group, for example an acetoxy group, or of a suitable etherified hydroxycarbonyl-oxy group.

/~

~._, ~)SS9~4 In a starting material of the formula III, the amino group is advc~ntageously protected by any of the, preferably easily removable , amino protective groups known in peptide chemistry or in penicillin and cephalosporin chemistry.
Such protective groups can be, for example, acyl, arylmethyl, 2-carbonyl-1-vinyl, arylthio or aryl-lower alkylthio groups and also arylsulphonyl groups as well as organic silyl or stannyl groups. The amino group can furthermore be in the form of the azido group. The starting material of the for-mula III can also be used in the form of an acid addition salt in which the amino group is pro-tected in the ionic form.
An easily removable acyl group is, for example, the formyl group or the acyl radical of a half-ester of carbonic acid, such as a lower alkoxycarkonyl group which, preferably at the carbon atom in the a-position to the oxy group, has several aliphatic substituents or is branched and/or aromati-cally or hetero-aromatically substi-tuted, or a methoxycarbonyl group which is substituted by an arylcarbonyl radical, especi-ally a benzoyl radical, or a lower alkoxycarbonyl group which is substituted by halogen in the ~-position, such as tert.-lower alkoxycarbonyl, ~or example tert.-butoxycarbonyl or tert.-pentoxycarbonyl , arylcarbonylmethoxycarbonyl, for example phenacyloxycarbonyl, 2-halogenoethoxycarbonyl, for example 2,2,2-trichloroethoxycarbonyl or 2-iodoethoxycarbonyl or a group which can be converted to the latter~ such as 2-chloroethoxycarbonyl or 2-bromoethoxycarbonyl, ard also preferably polycyclic cycloalkoxycarbonyl, for example adam-antyloxycarbonyl, phenyl-lower alkoxycarbonyl, in particular /~
_ ~ _ ~V55~

a-phenyl-lower alkoxycarbonyl, which is optionally substitu-ted, for example by lower alkyl 9 such as tert.-butyl, hydroxyl, lower alkoxy~ such as methoxy, and/or nitro,f~ example4- methoxy-benzyloxycarb~nyl, 4-hydroxy-3,5-bis-tert.-butyl-benzyloxy-carbonyl, 4-nitrobenzyloxycarbonyl or ~-4-biphenylyl-a-methyl-ethyloxycarbonyl and also diphenylmethoxycarbonyl which is optionally substituted, for example by lower aIkoxyj such as methoxy, for example diphenylmethoxycarbonyl, or furyl-lower alkoxycarbonyl, above all ~-furyl-lower alkoxycarbonyl, for example furfuryloxycarbonyl. An acyl group for protecting -the amino group can also be the corresponding radical of a suitable carboxylic acid such as of an aryldicarboxylic acid, for example the phthaloyl radical, or of ahalogeno- l~er alk-anecarboxylic acid, for example the trifluoroacetyl radical.
The following should be mentioned as examples of eas ly removable arylmethyl groups: optionally substituted polyarylmethyl groups, such as diarylme-thyl or triarylmethyl groups, for example trityl which is optionally substituted, such as by lower alkyl, for example methyl, and/or lower alkoxy, such as methoxy, in particular optionally o-methoxy-substituted or p-methoxy-substituted trityl.
Easily removable 2-carbonyl-1-vinyl groups which together with the amino group form either an enamine or the ke-timine tautomeric therewith are, for example, 2-lower alkoxycarbonyl-l-lower alkylvinyl groups, especially the 2- --methoxycarbonyl-l-methyl-l-vinyl group~
Easily removable arylthio or aryl-lower alkylthio groups are, for example, substituted, for example nitro-,~ /1 1()55~

substituted or halogen-substituted, for example chlorine-substituted, phenylthio groups, such as the 2-nitrophenyl-thio, the 2,4-dinitrophenylthio or the pentachlorophenylthio group, and al,o triarylmethylthio groups, for example the triphenylmethyl-thio group.
An easily removable organic silyl or stannyl group can preferably carry, as substituents, optionally substitu-ted, in particular aliphatic, hydrocarbon radicals, such as lower alkyl, for example methyl, ethyl or tert.-bu-cyl, or halogeno-lower alkyl, for example 2-chloroethyl, and also functional groups, for example etherified or esterified hydroxyl groups, such as lower alkoxy, for example methoxy or ethoxy, or halogen, for example chlorine. Such silyl or stannyl radicals are, inter alia, tri-lower alkylsilyl, for example trimethylsilyl or tert.-butyldimethylsilyl, lower alkoxy-lower alkyl-halogenosilyl, for example chloro-methoxy-methyl-silyl,or tri-lower alkylstannyl, for example tri-n-butyl-stannyl.
The acylation of the amino group, which is free or substituted by a radical which permits acylation, can be effectedin aknown manner, by treatment with an acid of the formula III or a reactive functional derivative thereof.
If a free acid of the formula III, preferably having a protected amino group~ is employed for the acylation, suitable condensation agents are usually employed, such as carbodiimides, for example N,N'-diethyl-, N,N'-dipropyl-, N,N'-diisopropyl-, N,N'-dicyclohexyl- or N-ethyl-N'-3-di-methylaminopropyl-carbodiimide, suitable carbonyl compounds, A~ ' --,~7 --.~ ,~' l~)5S~;~4 for example carbonyldiimidazole or isoxazolinium salts, for example N-ethyl-5-phenyl-isoxazolinium-3'-sulphonate and N-tert.-butyl-5-methyl-isoxazolinium perchlorate, or an acyl-amino compound, for example 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline.
The condensation reaction is preferably carried out in an anhydrous reaction medium, preferably in the presence of a solvent or diluent, for example methylene chloride, dimethylformamide or acetonitrile, if desired or required with cooling or warming and/or in an inert gas atmosphere.
An amide-forming functional derivative of an acid of the formula III, preferably having a protected amino group, is above all an anhydride of such an acid, including a mixed anhydride, the latter being preferred, but also an inner anhydride, that is to say the corresponding ketene. Mixed anhydrides are, for example, those with inorganic acids, such as hydrogen halide acids, that is to say the corresponding acid halides, for example chlorides or bromides, and also with hydrazoic acid, tha-t is to say the corresponding acid azides~
with an acid containing phosphorus, for example phosphoric acid or phosphorous acid, or with an acid containing sulphur, for example sulphuric acid, or with hydrocyanic acid, Further mixed anhydrides are, for example, those with organic carboxy-lic 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 trichloro-acetic acid, or with half-esters, in particular lower alkyl half-esters of carbonic acid~such as the ethyl half-ester or iso-/q ¢~

1l~5S~

butyl half-ester of carbonic acid, or with organic, in par-ticular aliphatic or aromatic, sulphonic acids, for example p-toluenesulphonic acid.
Furth~r acid derivatives of an acid of the formula III which are suitable for reaction with the amino group are activated esters, usually with a protected amino group in the aminomethyl grouping, such as esters with vinylogous alcohols (that is to say enols), such as vinylogous lower alkenols, or aryl esters, such as 4-nitrophenyl or 2,4-dinitrophenyl esters, heteroaromatic esters, such as benztriazole esters, for example 2-benztriazole esters, or diacylimino esters, such as succinylimino es~ers or phthalylimino esters.
The acylation with an acid derivative, such as an anhydride and in particular 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 trimethylamine, triethylamine or ethyldiisopropylamine, or a N,N-di-lower alkylaniline, for example N,N-dimethylaniline, ~r a base of the pyridine type, for example pyridine, an inorganic base, for example an alkali metal hydroxide,carbonate or bicarbonate or alkaline earth metal hydroxide, carbonate or bicarbonate, for example sodium hydroxide, carbonate or bi-carbonate, 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.
me above acylation can be carried out in an inert, ~0 ~ ~

1(~5S~
preferably anhydrous, solvent or solvent mixture, for example in a carboxylic acid amide, such as a forrnamide, for example dimethylformamide, a halogenated hydrocarbon, for example methylene chloride, carbon tetrachloride or chlorobenzene, à ketone, for example acetone, an ester, for example ethyl acetate, or a nitrile, for example acetonitrile, or mixtures thereof and~ if necessary, at lowered or elevated temperature and/or in an inert gas atmosphere, for example a nitrogen atmosphere.
In an acid of the formula III or in an acid derivative thereof, the amino group is usually in a protected form, and in a starting ma-terial of the formula III the protected amino group can also be in an ionic form, that is to say the starting material of the formula III is used in the form of an acid addition salt, preferably with a strong inorganic acid, such as a hydrogen halide acid, for example hydrochloric acid, or sulphuric acid.
Furthermore an acid deriva-~ive can, if desired, be formed in situ. Thus, for example, a mixed anhydride is obtained by treating an acid of the formula III~or a suitable salt thereof, such as an ammonium salt, for example with an organic amine, such as 4-methylmorpholine, or a metal salt, for example an alkali metal salt, with a suitable acid derivative, such as a corresponding acid halide of an option-ally substituted lower alkanecarboxylic acid~ for example trichloroacetyl chloride, or with a half-ester of a carbonic acid half-halide, for example chloroformic acid ethyl ester or isobutyl ester, and the mixed anhydride thus obtainable is ~1 --3~ ~

5~J~ ~

used without isolation.
In a resulting compound it is possible, if desired or required, to convert functional optionally protected groups into other functional, for ; example free functional, groups in a manner which is in itself known. Above all, in a compound obtainable according to the invention, a protected amino group in the aminomethyl substituent of the acylamino grouping has to be liber-ated and/or a protected carboxyl group of the formula -C(=0)-Ro which differs from a carboxyl grouping of the formula -C(=0)-R has to be converted into a group of the formula -C(=0)-R; furthermore it is possible, if desired, to con-vert a free carboxyl group of the formula -C(=0)-R in a manner which is in it-self known into a physiologically splittable carboxyl group of the formula -C(=0)-R and/or to convert a group Rl in a grouping of the formula Ib into another group Rl. These conversions are carried out in a manner which is in itself known, the sequence in multiple conversions being optional and usually depending on the nature of the radicals to be converted or to be split off and on the reactions employed for the purpose. Furthermore it is possible to convert more than one protected functional group simultaneously into the corres-ponding free functiQnal groups. Thus, for example, it is possible, by treat-ment with a suitable acid, such as trifluoroacetic acid, optionally in the presence of anisole, simultaneously to convert, in a resulting compound, a tert.-butoxycarbonylamino or diphenylmethoxycarbonylamino group in the amino-ethyl substituent of the acylamino radical in the 7-position and a diphenyl-methoxycarbonyl group representing the radical of the formula -C(=0)-Ro in the 4-position of a resulting 3-cephem compound simultaneously into the amino and carboxyl group respectively.
The conversion of a protected amino group into a free amino group can be effected in a manner which is in itself known, usually by solvolysis or re-duction.
A formyl group used as an amino protective group can be split off, for example by treatment with an acid agent, for example p-toluenesulphonic acid or hydrochloric acid, with a weakly basic agent, for example dilute = onia, or with a decarbonylating agent, for example tris-(triphenylphosphine)-~ -22-~ ,. .. ..

~)S59i~4 rhodium chloride.
In a resulting compound, an easily removable acyl group, such as an ~-poly-branched lower alkoxycarbonyl group, for example tert.-butoxycarbonyl, and also a polycyclic cycloalkoxycarbonyl group, for example l-adamantyloxy-carbonyl, an optionally substituted diphenylmethoxycarbonyl group, for example diphenylmethoxycarbonyl, or an ~-furyl-lower alkoxycarbonyl group, can be re-moved from an acylamino group, for example by acidolysis, such as treatment with a suitable acid, such as a strong, preferably aliphatic, carboxylic acid, for example an optionally halogenated, in particular fluorinated, lower alkane-carboxylic acid, above all formic acid or trifluoroacetic acid, optionally inthe presence of a nucleophilic reagent, such as anisole, and a formyl group can be removed by treatment with a strong acid, such as a mineral acid, for example hydrochloric acid, or a strong organic ~ 23 l~S~g~
i sulphonic acid, for example 4-methylphenylsulphonic acid, and also by treatment with a decarbonylating agent, for example tris-triphenylphosphine-rhodium chloride, whilst, for example, r suitably substituted benzyloxycarbonyl group, such as 4-hydroxy-3,5-bis-tert.-butyl-benzyloxycarbonyl, can be removed, for example, by treatment with an optionally anhydrous, weak base, such as an alkali metal salt of an organic carboxylic acid, for example the sodium salt or potassium salt of 2-ethyl-pentanecarboxylic acid, with an alkali metal salt of a thiophenol, for example the sodium salt of thiophenol, or with a suitable organic amine, for example ethylamine or cyclohexylamine, or a suitably sub-stituted lower alkanoyl group, for example trifluoroacetyl, can be removed by hydrolysis under weakly basic conditions.
A 2-halogeno-lower alkoxycarbonyl group, such as 2,2,2-trichloroethoxycarbonyl or 2-iodoethoxycarbonyl, or a phen-acyloxycarbonyl group, can be removed, for example, by treat-ment with a chemical reducing agent, such as a suitable reducing metal or a corresponding metal compound, for example zinc, or a chromium-II compound, such as chromium-II chloride or chromium-II acetate, usually in the presence of an agent which together with the metal or the metal compound produces nascent hydrogen, preferably in the presence of aqueous acetic acid. A phenacyloxycarbonyl group can also be replaced by hydrogen by treatment with a suitable nucleo-philic, preferably salt-forming, reagent, such as sodium thiophenolate.
Furthermore, in a resulting compound an amino group ~C ~ -- --,,33 ~

lOS55~;~4 protected by a suitably substituted benzyloxycarbonyl group, such as 4-methoxy- or 4-nitro-benzyloxycarbonylamino can be split reductively, such as hydrogenolytically, for example by treatment with hydrogen in the presence of a hydrogenation catalyst, for example palladium9 and, in particular in the case of 4-nitrobenzyloxycarbonylamino, by treatment with a chemical reducing agent, for example sodium dithionite.
A polyarylmethyl group, such as the trityl group, can be split off by, for example, treatment with an acid agent, such as a mineral acid, for example hydrochloric acid.
An amino group protected in the form of an enamine or of a ketimine tautomeric therewith, as well as an amino group protected by arylthio, aryl-lower alkylthio or arylsulphonyl, can be split, for example, by treatment with an acid agent, above all an aqueous acid, such as an organic carboxylic acid, for example formic acid, acetic acid or propionic acid, or a mineral acid, for example hydrochloric acid or sulphuric acid, optionally in the presence of a water-miscible solven-t, such as a lower alkanol, for example methanol, a ketone, for example acetone, an elher, for example tetrahydrofurane, or a nitrile, for example acetonitrile. m e removal of the said thio protective groups can be effected particularly rapidly in the presence of additional reagents, such as sodium thio-sulphate, sulphurous acid, thioacetamide, thiourea and potas-sium iodide.
An amino group, protected wlth an organic silyl or stannyl group, in a resulting compound can be liberated by treatment with an aqueous or alcoholic agent, for example with Z~ .

~t)SS~

a lower alkanol, such as methanol, or a mixture thereof;
usually, the split-ting of an amino group protected in this way takes place already during working up of the acylation product.
An amino group, present in the form of an azido group, in the aminomethyl substituent of a resulting compound can be converted to the amino group in a manner which is in itself known by means of reduction, for example by treatment with hydrogen in -the presence of a hydrogenation catalyst, such as a nickel catalyst or palladium catalyst, for example in the presence Cf Raney nickel or palladium on charcoal, under mild conditions, for example ~nder atmospheric pressure and/or at room temperature or only slightly elevated tempera-ture, and also by treatment with a phosphine, such as a triarylphosphine, for example triphenylphosphine, or with tin-II chloride.
The reaction products which result from the acylation, according to the invention, of compounds of thè formula II, wherein the amino group is substituted by a silyl or stannyl group, and in which the organic silyl or stannyl group is still present on the amide nitrogen are usually converted into compounds of the formula I during working up, in particular under hydrolytic and/or alcoholytic conditions, for example such as are customary when splitting off organic silyl or stannyl groups from amino groups.
The reaction products which result from the acylation, -according to the invention, of compounds of -the formula II, wherein the amino group is substituted by an ylidene group~

; 2,6 ~)S~4 are also usually converted into compounds of the formula I
during working up, in particular by hydrolysis, for example by treatment with water.
In a compound of the formula I obtainable according to the invention, having a protected, in particular esteri-fied, carboxyl group of the formula -C(=0)-Ro, the latter can be converted into -the free carboxyl group in a manner which is in itself kno~, for example by solvolysis, treat-ment with a nucleophilic reagent, irradiation or reduction, depending on the nature of the group Ro~ A carboxyl group which is esterified by a suitable 2-halogeno-lower alkyl group, such as 2,2,2-trichloroethyl or 2-iodoethyl, or an arylcarbonylmethyl group, such as phenacyl, can be split, 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 acetate, usually in the presence of a hydrogen donor~ which together with the metal is able to generate nascent hydrogen, such as an acid, above all acetic acid and also formic acid, preferably with the addition of water; a carboxyl group esterified by an arylcarbonyl group, for example a phenacyl group, can also be converted into the free carboxyl group by treatment with a nucleophilic, preferably salt-forming, reagent, such as sodium thiophenolate or sodium iodide. A carboxyl group esterified by a suitably substitu-ted arylmethyl group can, for example, be converted into the free carboxyl group by irradiation, preferably with ultraviolet light, for example below 290 m~ if the arylmethyl group represents, for example, _ ~ _ i~s~

a benzyl radical which is optionally substituted in the 3-, 4- and/or 5-position, for example by lower alkoxy groups and/or nitro groups, or with ultraviolet light of longer wavelengths, for example above 290 m~, when the arylmethyl group denotes, for example, a benzyl radical substituted by a nitro group in the 2-position. The carboxyl group can be liberated from a carboxyl group esterified with a suitably branched lower alkyl group, for example tert.-butyl, with a suitable cycloalkyl group, such as l-adamantyl, or with a diphenylmethyl group, for example benzhydryl, for example by treatment with a suitable acid agent, such as formic acid or - trifluoroacetic acid, optionally with addition of a nucleo-philic reagent, such as phenol or anisole. An esterified carboxyl group which can be split hydrolytically, such as a carboxyl group esterified by a suitably substituted phenyl radical or a diacyliminomethyl radical, and also a carboxyl group esterified with the 4-hydroxy-3,5-di-ter-t.-butyl-benzyl radical, can, depending on the nature of the ester grouping, be split, for example, by treatment with an acid or weakly basic aqueous agent, such as hydrochloric acid or aqueous sodium bicarbonate or an aqueous potassium phos-phate buffer of pH about 7 to about 9, whilst an esterified carboxyl group which can be split hydrogenolytically, such as an a-aryl lower alkyl group which is optionally substituted in the aryl radical, for example benzyl, 4-methoxy-benzyl or 4-nitrobenzyl, can be split by hydrogenolysis, for example by treatment with hydrogen in the presence of a noble metal cata-lyst, for example a palladium catalyst.

,,~., - ~_ ~5$~4 A carboxyl group protected, for example, by silylation or stannylati-on and also by phosphorylation can be liberated in the usual manner, for ex-ample by hydrolysis or alcoholysis.
The new compounds of the formula I can also be obtained if the meth-oxy group is introduced into the 7~-position of a 3-cephem compound of the formula H2N-CH2 ~ ~ ~ ~ S`CIH2 (IV) X N`C ~C-R

~ C~

wherein amino of the aminomethyl group is preferably in the protected form, a carboxyl group of the formula -C(=0)-Ro preferably being present in a pro-tected form, or of a salt thereof, and, if desired or required, the addition-al process steps are carried out.
The introduction of the methoxy group into the 7~ position of a 3-cephem starting material of the formula IV can be carried out in a manner which is in itself known.
Thus, an acylimino compound of the formula O H
H2N_CH2 t~ CH2-C N ~ ~,- ~CH2 0=C-R

wherein amino of the aminomethyl group is present in a protected form and a carboxyl group of the formula -C(=0)-Ro is present in a protected form, can be treated with methanol and in a resulting compound protected amino of the amino-methyl group can be converted into free amino and, if required or desired, a f~
~, ~ss~
carboxyl group of the formula -C(=0)-R can be converted into a carboxyl group of the formula -C(=0)-R and/or, if desired, a group Rl can be converted into another group Rl and/or, if desired, a resulting salt can be converted into the free compound or into another salt, or a resulting free compound can be converted into a salt.
In the starting material of the formula IVa, free functional groups, in particular amino of the aminomethyl substituent and a carboxyl group -C(=0)-Ro and also free functional groups optionally present in a radical R2, are pre-sent in a protected form, for example in a form protected as indicated above, an amino group, for example, as an acylamino group which can preferably be split easily, and also as an arylmethylamino, 2-carbonyl-1-vinyl-amino, aryl-thioamino or aryl-lower alkylthioamino group, a carboxyl group, for example, as an esterified carboxyl group which preferably can be split easily, and a functional group in the radical R2, such as, for example, a hydroxyl group, in a form which is protected as indicated above, for example in the form of an acyloxy group.
The above reaction is carried out in a manner which is in itself known, usually in the presence of a solvent or diluent or of a mixture of these, it being possible for methanol at the same time also to serve as the solvent or diluent, preferably with cooling, for example down to about -80C, and also at room temperature or with slight warming, ant, if necessary, in a closed ves-sel and/or under an inert gas atmosphere, for example a nitrogen atmosphere.
The starting material of the formula IVa is usually reacted in the crude form, that is to say without being isolated after its manufacture, with the methanol, or is formed in the presence of the latter reagent. Here, for example, the starting material is a compound of the formula IV, wherein amino of the aminomethyl group and the carboxyl group of the formula -C~=~)-Ro, as well as optionally additionally present functional groups, are present in a protected form, and is treated with an anion-forming agent, followed by an N-halogenating agent, and then reacted, if necessary, with a base which splits off hydrogen halide; alternatively, a compound of the formula ~ . -30-. . ~, . . .

l~S~

H2N-CH2 ~ CH2-C-HN ~ S ~Cl2 (IVb) X O N

Ro 0 wherein R represents an organic radical and wherein amino of the aminomethyl group and the carboxyl group of the formula -C(=0)-Ro, and optionally addition-ally present functional groups, are present in a protected form, is reacted with halogen, followed by a base. It is thus possible to obtain, as a product which is usually not isolated, the corresponding 7-acylimino-3-cephem compound of the formula IVa which in the presence of methanol is converted into the desired 7 ~acylamino-7a-methoxy-3-cephem compound, on which the above-mention-ed additional steps are carried out if required or desired.
A suitable anion-forming agent with which a starting material of the formula IV i5 reacted is above all an organo-metallic base, in particular an organo-alkali metal base, above all an organo-lithium base. Such compounds are, in particular, corresponding alcoholates, such as suitable lithium lower alkanolates, above all lithium methylate, or corresponding metal~hydrocarbon bases, in particular lithium-lower alkanes and preferably lithium-phenyl. The reactlon with the anion-forming organo-metallic base is usually carried out with cooling, for example at about 0C to about -80C, and in the presence of a suitable solvent or diluent, for example an ether, such as tetrahydrofurane, or also in the presence of methanol when using lithium methylate, and, if desired, in a closed vessel and/or in an inert gas atmosphere, for example a nitrogen atmosphere.
The N-halogena~ing agent employed is usually a sterically hindered organic hypohalite, in particular hypochlorite, and above all a corresponding aliphatic hypohalite, for example hypochlorite, such as a tert.-lower alkyl hypohalite, for example hypochlorite. Above all, tert.-butyl hypochlorite is employed and is reacted with the non-isolated product of the anionisation reaction.

~ ~ -31-lVSS~

The N-halogenated intermediate compound is converted, if an excess of the anion-forming base, especially of lithium methylate, is present under the reaction conditions, and without being isolated, into the acylimino com-pound of the formula IVa, and if methanol is present this compound is directly converted into the 7~-methoxy-3-cephem compound. If necessary, the elements of hydrogen halide, especially of hydrogen chloride, must be eliminated from the N-halogenated intermediate product; this is done by adding a base which eliminates hydrogen halide, such as a suitable alkali metal lower alkanolate, for example lithium tert.-butylate, and this reaction usually takes place un-der the conditions of the anion-forming and N-halogen compound-forming reaction and can be carried out in the presence of methanol, in which case, instead of the acylimino compound, the 7-methoxy-3-cephem compound can be obtained dir-ectly. In this case, a compound of the formula IV wherein functional groups are usually present in a protected form is used as a starting material and is reacted with an excess of the anion-forming agent, for example lithium methy-late or phenyl-lithium, in the presence of methanol, the product is then treat-ed with the N-halogenating agent, for example tert.-butyl hypochlorite, and the desired compound of the formula I is thus obtained direct: in this com-pound, protected functional groups can be liberated if required or desired.
Alternatively, the methanol can be added subsequently, in which case the de-hydrohalogenation and the addition of methanol can be carried out at somewhat higher temperatures than the anion-forming and N-halogen compound-forming re-actions, for example at about 0C to about -20C, if necessary in a closed vessel and/or in an inert gas atmosphere, for example a nitrogen atmosphere.
In a starting material of the formula IVb, an organic radical R
above all denotes a hydrocarbon radical of aliphatic character, such as lower alkyl and especially methyl. The reaction with halogen, above all with chlor-ine, and a base is usually carried out in the presence of a suitable solvent or diluent, such as a halogenated hydrocarbon, for example methylene chloride, and with cooling, for example down to about -80C, and can, if necessary, be carried out in a closed vessel and/or in an inert gas atmosphere, for example a nitrogen atmosphere. The base used is preferably a suitable organic base, -~ i -32-1l)55~
such as a tertiary amine, for example a tri-lower alkylamine, such as tri-ethylamine, which is usually added to the above halogenation mixture together with methanol and under the conditions of the halogenation process. In this way, the 7-acylimino-3-cephem starting material of the formula IV is converted direct into a 7 ~acylamino-7a-methoxy-3-cephem compound of the formula I which can, if required or desired, be converted into the desired compound of the formula I.
The introduction of the methoxy group into the 7a-position of 7 ~
acylamino-3-cephem compounds can also be effected by replacement of a replace-able group present in this position. Thus, a compound of the formula IVb,wherein R

~ -33-has the indicated meaning, and above all represents an ali-phatic hydrocarbon radical, such as lower alkyl and especially methyl, and wherein amino of the aminomethyl group and the carboxyl group of the formula -C(=O)-Ro~ as well as optionally additionally present functional groups, are present in a pro-tected form, can be reacted with methanol in the presence of a desulphurisation agent.
The desulphurisation in the presence of methanol is usually effected using a suitable silver compound or mercury compound, such as silver oxide or mercury oxide or, in par-ticular, a corresponding salt, such as a silver-I salt or mercury-II salt with an organic carboxy]ic acid, for example a silver-I lower alkanoate or mercury-II lower alkanoate, in particular mercury-II acetate. The reaction is carried out in the presence of a solvent or diluent, for example an ether, such as dimethoxyethane, or a solvent mixture, it also being possible to use an excess of methanol as -the latter, with cooling, for example down to about -30C, at room temperature or with slight warming, for example up to about +70C, if necessary in a Glosed vessel and/or in an inert gas atmos-phere, for example a nitrogen atmosphere.
In the compounds obtainable according to the variants of the above methoxylation process, which are preferably carried out in accordance with the methods described by . . .
Koppel and Kocher, J. Am. Chem. Soc., volume 95, page 2,403 (1973), Spitzer and Goodson, Tetrahedron Letters, page 273 (1973) and Slusarchyk et al., J. Org. Chem., volume 38, page 943 (1973), the protected amino of the aminomethyl group is , 3 ~ -_ ,~ _ ' lOS~9Z~
liberated in accordance with the abovementioned processes and, if necessary, a protected carboxyl group of the formula -C(=0)-R is converted into a carboxyl group of the formula -C(=0~-R in accordance with the process mentioned; if desired it is possible, as mentioned, to convert, in a result-ing compound, the carboxyl group of the formula -C(=O)-R into another carboxyl group of the formula -C(=O)-R and/or a group Rl into another group Rl.
The compounds of the present invention can also be obtained by a method wherein, in a 3-cephem compound of the formula O OCH3 H
H2N CH2 ~ CH2 -C _ N ~ S~ ~CH

X O=C ~C '~ 1 ( V), j R~ ~O
o CH2-- C~' Ro wherein Am represents a protected amino group and Rl denotes a radical which together with the carbonyl grouping of the formula -C(=O)-forms a preferably protected carboxyl group, and wherein amino in the aminomethyl group is present in a protected form, which differs~ in its method of conversion into the free amino group, from that of the protected amino group AmO~ the group Am is converted into the free amino group and the 5-amino-5-carboxy-valeryl radical is split off under the reaction conditions, and in a resulting compound protected amino of the aminomethyl group is converted into 5~'~4 free amino and, i~ required or desired, a carboxyl group of the formula -C(=O)-R is converted into a carboxyl group of the formula -C(=O)-R and/or, if de-sired, a group Rl is converted into another group Rl and/or, if desired, a re-sulting salt is converted into the free compound or into another salt, or a resulting free compound is converted into a salt.
In the above-mentioned starting material, the carboxyl groups of the formula -C(=O)-Ro and of the formula -C(=O)-Ro, represent carboxyl groups protected in the usual manner, such as, for example, the above-mentioned pro-tected carboxyl groups, and a group of the formula -C(=O)-Ro can also be an esterified carboxyl group which cannot be split, such as, for example, methoxy-carbonyl. Amino of the aminomethyl group is, as already noted, present in a protected form; usually, functional groups present in a radical Rl are also protected, for example as indicated. Groups protected in this way are usually not liberated under the reaction conditions.
A protected amino group AmO is usually a corresponding amino group which can be converted into the free amino group, preferably under mild con-ditions. Examples of protective groups are acyl, arylmethyl, 2-carbonyl-1-vinyl, arylthio, aryl-lower alkylthio or arylsulphonyl groups, which can be split off in various ways.
An acyl group which can be split off easily is, for example, the formyl group or the acyl radical of a half-ester of carbonic acid, such as a lower alkoxycarbonyl group which, preferably at the carbon atom in the ~-posi-tion to the oxy .~ -36-~l~5S~4 .
group7carries several aliphatic substituents or is branched and/or is aromatically or heteroaromatically substituted, or a methoxycarbonyl group substituted by an arylcarbonyl radi-cal, especial'y by a benzoyl radical, or a lower alkoxycar-bonyl group which is substituted by halogen in the ~-position, such as tert.-lower alkoxycarbonyl, for example tert.-butoxy-carbonyl, or -tert.-pentoxycarbonyl , arylcarbonylmethoxy-carbonyl, for example phenacyloxycarbonyl, 2-halogenoethoxy-carbonyl, for example 2,2,2-trichloroethoxycarbonyl or 2-iodoethoxycarbonyl or a group which can be converted into the latter, such as 2-chloroethoxycarbonyl or 2-bromoethoxycar-bonyl and also preferably polycyclic cycloalkoxycarbonyl, for example adamantyloxycarbonyl, phenyl-lower alkoxycarbonyl, in particular a-phenyl-lower alkoxycarbonyl which is optionally substituted, for example by lower alkyl 9 such as tert.-butyl, hydroxyl, lower alkoxy, such as methoxy, and/or nitro, for example 4-methoxy-benzyloxycarbonyl~ 4-hydroxy-3,5-bis-tert.-butyl-benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl or ~_L~
biphenylyl-u-methylethyloxycarbonyl and also diphenylmethoxy-carbonyl which is optionally substituted, for example by lower alkoxy, such as methoxy, for example diphenylmethoxycarbonyl~
or furyl-lower alkoxycarbonyl, above all ~-furyl-lower alkoxy-carbonyl, for example furfuryloxycarbonyl. An acyl group can also be the corresponding radical of a suitable carboxylic acid, such as of an aryldicarboxylic acid, for example the phthaloyl radical, orof a halogenD-lower alkanecarboxylic acid, for example the trifluoroacetyl radical.
me following should be mentioned as examples of easily t ,C, ,~

l~SS~4 removable arylme-thyl groups: optionally substituted polyaryl-methyl groups, such as diarylMethyl or triarylmethyl groups, for example trityl which is optionally substitu-ted, such as by lower alkoxy, such as methoxy, in particular optionally o-methoxy-substituted and/or p-methoxy-substituted trityl.
Easily removable 2-carbonyl-1-vinyl groups which together with an amino group form either an enamine or the ketimine tautomeric therewith are, for example, 2-lower alkoxycarbonyl-l-lower alkylvinyl groups, especial]y the 2-methoxycarbonyl-l-methyl-l-vinyl group.
Easily removable arylthio or aryl-lower alkylthio groups are, for example, substituted, for example nitro-substituted or halogen-substituted, for example chlorine-substituted, phenylthio groups, such as the 2-nitrophenylthio, the 2,4-dinitrophenylthio or the pentachlorophenylthio group, and also triarylmethylthio groups, for example the triphenyl-methylthio group.
An amino group AmO protected in this way can be con-verted into the free amino group in a manner which is in itself known; the amino group liberated effects the intra-molecular aminolysis, which takes place under the reaction conditions~ of the 5-amino-5-carboxy-valeroyl radical, which is split off and is then usually present in the form of the protected 2-oxo-piperidine-6-carboxylic acid.
The splitting of a protected amino group AmO to give a free amino group can be carried out in various ways which are in themselves known and depend on the nature of the pro-tective group, in particular by solvolysis, treatment with a _3g ~V~S~4 nucleophilic reagent or reduction.
A formylamino group AmO can be split, for example, by treatment with an acid agent, for example p-toluenesulphonic acid or hydro~hloric acid, with a weakly basic agent, for example dilute ammonia, or with a decarbonylating agent, for example tris-(triphenylphosphine)-rhodium chloride.
An a-poly-branched lower alkoxycarbonylamino group, for example tert.-butoxycarbonylamino, and also a polycyclic cycloalkoxycarbonylamino groupy for example l-adamantyloxy-carbonylamino, an optionally substituted diphenylmethoxy-carbonylamino group, for example diphenylmethoxycarbonyl-amino, or an a-furyl-lower alkoxycarbonylamino group AmO can be split, for example, by treatment with a suitable acid, such as a strong, preferably aliphatic, carboxylic acid, such as an optionally halogenated, in particular fluorinated, lower alkanecarboxylic acid~ above all formic acid or trifluoro-acetic acid, optionally in the presence of a nucleophilic reagent, for example anisole, whilst a sui-tably substituted benzyloxycarbonylamino group, for example 4-hydroxy-3,5-di-tert.-butyl-benzyloxycarbonylamino can preferably be split by treatment with an optionally anhydrous weak base, such as an alkali metal salt of an organic carboxylic acid, for example the sodium salt or potassium salt of 2-ethyl-pentanecarboxylic acid, with an alkali metal salt of a thiophenol, for example the sodium salt of thiophenol, or with a suitable organic amine, for example ethylamine or cyclohexylamine, whilst a suitably substituted lower alkanoylamino group, for example trifluoroacetylamino, can be split hydrolytically under weakly _ ,~ _ .~

l~S~

basic conditions. A 2-halogeno-lower alkoxycarbonylamino group, such as 2,2,2-tr].chloroethoxycarbonylamino or 2-iodo-ethoxycarbonylamino (a group which can be converted into 2-iodoethoxycarbonylamino, such as the corresponding 2-chloro-ethoxycarbonylamino or 2~bromoethoxycarbonylamino, being .
converted, kefore splitting off, into 2-iodoethoxycarbonyl-amino in a mal~ner which is in itself known, for example by treatment with a suitable iodine salt, such as an alkali metal iodide, such as sodium iodide, in the presence of a solvent, such as acetone), or a phenacyloxycarbonylamino group, such as phenacyloxycarbonylamino, can be split by treatment with a chemical reducing agent, such as a suitable reducing metal or a corresponding metal compound, for example zinc, or a chromium-IL compound, such as chromium-II chloride or chromium-II acetate, usually in the presence of an agent which together with the metal or the metal compound generates nascent hydrogen, preferably in the presence of aqueous acetic acid.
It is also possible to split an amino group AmO which is protected by a preferably suitably subs~ituted benzyloxycarbonyl group, such as 4-methoxy- or 4-nitro-benzyloxycarbonylamino, hydrogenolytically, for example by treatment with hydrogen in the presence of a hydrogenation catalyst, for example pal-ladium, or, in particula~ 4-nitrobenzyloxycarbonylamino, by treatment with a chemical reducing agent, for example sodium dithionite.
A polyarylmethylamino group AmO, such as tritylamino, can be split, for example, by treatment with an acid agent, ~0 --,5~ --.

1~ )55~ ~L/~

such as a mineral acid, for example hydrochloric acid.
An amino group pro-tected in the form of an enamine or of a ketimine tautomeric therewith, and the abovementioned amino groups AmO protected by arylthio, aryi-lower alkylthio or arylsulphonyl can be split, for example~ by treatment with an acid agent, above all an aqueous acid, such as an organic carboxylic acid, for example formic acid, acetic acid or propionic acid, or a mineral acid, for example hydrochloric acid or sulphuric acid, optionally in the presence of a water-miscible solvent, such as a lower alkanol, for example methanol, a ketone, for example acetone, an ether, for example tetrahydrofurane, or a nitrile, for example acetoni-trile.
me splitting off of the thio protected groups mentioned can take place particularly rapidly in the presence of additional reagents, such as sodium thiosulphate, sulphurous acid, thio-acetamide, thiourea and potassium iodide.
The splitting reactions described above are carried out under conditions which are in themselves known, if neces-sary with cooling or warming, in a closed vessel and/or in an inert gas atmosphere, for example a nitrogen atmosphere.
As mentioned above, simultaneous liberation of other protected functional groups present in the starting material should not take place under the reaction conditions. Thus, for example, the protected amino group AmO can be an amino group which can be split on treatment witn a chemical reducing agent, for example with zinc in the presence of aqueous acetic acid, for example a 2-halogeno-lower alkoxycarbonylamino group which can be split under these condi-tions, such as 2,2,2-tri-¢ !

~chloroethoxycarbonylamino, whilst amino of the aminomethyl substituent can be protected, for example, by a lower alkoxy-carbonyl group which can be split off on treatment with a suitable acid, such as trifluoroacetic acid, for example by an a-poly-branched lower alkoxycarbonyl group, such as -tert.-butoxycarbonyl, and a carboxyl group of the formula -C(=0)-R
and also a free carboxyl group optionally present in a group R2 in the radical of the formula -S-Ao~, can be protected, for example, by a diphenylmethyl group which can also be split off on treatment with a suitable acid, such as trifluoroacetic acid, for example by an optionally substituted diphenylmethyl group, for example benzhydryl, and withstand the above-mentioned reductive splitting conditions when in the protected form, and are only liberated, if required or desired, after intramolecular aminolysis of a 5-amino-5-carboxy-valeryl radical has taken place.
The above process can be carried out, for example, according to the method described by Sle-tzinger et al., J. Am.
Chem. Soc., volume 94, page 1,410 (1972~.
m e compounds of the present invention can also be manufactured by a method wherein a compound of the formula C ~ 1 (~
0=C-R

wherein X and Rl have the above mentioned meaning and a carboxyl group of the formula .~

~()S5~4 -C(=O)-Ro is preferably present in a protected form, is reacted with a compound of the formula RX-NH2 (VII), wherein Rx denotes an amino protective group, and formaldehyde in the presence of a strong, at most slightly nucleophilic acid, and in a resulting compound protected amino of the aminomethyl group is converted into free amino and, if required or desired, a carboxyl group of the formula -C(=O)-Ro is conver-ted into a carboxyl group of the formula -C(=O)~R and/or, if desired, a group Rl is converted into another group Rl and/or, if desired, a resulting salt is converted into the free com-pound or into another salt~or a resulting free compound is converted into a salt.
An amino protective group Rx is one of the above-mentioned amino protective groups, mentioned, for example, in the context of amino of the aminomethyl radical, which cannot be split off ~mder the reaction conditions, that is to say in the presence of the strong, at most slightly nucleophilic, acid. Such a protective group is above all a corresponding acyl group, such as formyl or suitable op-tionally substituted lower alkanoyl, in particular trifluoroacetyl, and above all sui-tably etherified hydroxycarbonyl which can be split off, for example under reductive conditions, on treatment ~ith a nucleophilic reagent or on irradiation, above all 2-halogeno-lower alkoxycarbonyl, for example 2,2,2-trichloroethoxy-carbonyl, 2-chloroethoxycarbonyl, 2-bromoethoxycarbonyl or 2-iodoethoxycarbonyl, arylcarbonylmethoxycarbonyl, for example phenacyloxycarbonyl, or -aryl-lower alkoxycarbonyl, such as -phenyl-lower alkoxycarbonyl which is optionally substituted, ~3 --~ _ 1~)5~

for example by lower alkoxy, such as methoxy, and/or nitro, for example benzyloxycarbonyl, 4-me-thoxy-benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl or 4,5-dimethoxy-2-nitrobenzyloxy-carbonyl.
Formaldehyde can be employed as such or in the form of a reactive derivative thereof, above all in the form of a polymer, such as in the form of paraformaldehyde.
Strong, at most slightly nucleophilic acids are above all strong organic carboxylic acids, such as preferably halogen-substituted lower alkanecarboxylic acids, for example formic acid (optionally in -the presence of a strong organic sulphonic acid, such as a strong arylsulphonic acid, for example 4-methylbenzenesulphonic acid) and above all tri-fluoroacetic acid.
The above reaction is usually carried out by adding to a reaction mixture of a compound of the formula VII and formaldehyde or a derivative thereof (which is prepared, for example, in the presence of an iner~ solvent and of a weakly basic agent, such as an alkali me-tal carbonate, for example potassium carbonate, and, if necessary, with removal of water and formation of a compound of the formula RX-NH-CH2-OH (VIIa) obtainable as an intermediate product) the starting material of the formula VI and the strong, at most slightly nucleo-philic acid, the process being carried ou-t in the presence of an inert solvent or solvent mixture, with stirring or warming and/or in an inert gas atmosphere.
In a compound of the formula I obtainable according to this process 9 the amino group in the aminomethyl radical is : ~Y
'.._: J , _ ~ .~

105S~
present in a protected form, amino protective groups being above all the abovementioned acyl radicals. They are split off in a manner ~Yhich is in itself known, for example as des-cribed above, a formyl group, for example, by treatment with a strong acid, for example hydrochloric acid or 4-methyl-phenylsulphonic acid, a trifluoroacetyl group, for example, hydrolytically under weakly basic conditions, a suitable 2-halogeno-lower alkoxycarbonyl or arylcarbonylmethoxycarbonyl group, for example, by treatment with a chemical reducing agent, such as a suitable reducing metal or a corresponding metal compound, for example zinc, or a chromium-II compound, such as chromium-II chloride or chromium-II acetate, usually in the presence of an agent which together with the metal or the metal compound generates nascent hydrogen, preferably in the presence of aqueous acetic acid, an arylcarbonylmethoxy-carbonyl group also by treatment with a suitable nucleo-philic, preferably salt-forming, reagent, such as sodium thiophenolate, and an a-aryl-lower a.lkoxycarbonyl group hydrogenolytically, for example by treatment with hydrogen in the presence of a hydrogenation catalyst, for example pal-ladium, or, such as 4-nitrobenzyloxycarbonyl, by treatment with a chemical reducing agent, for example sodium dithionite.
The 3-cephem compounds of the present invention can also be obtained by a method in which a 2-cephem compound of the formula '7f~,' ., _ ,~g _ 11)55~4 o ll OCI~ M
H2~--Cll ~ X ~ CH2--~--H~~ 3 O Rl ~ (VIII) O=C - Ro wherein amino of the aminomethyl group and/or a carboxyl group of the formula--C(=O)-RO are present in a protected form, if required or desired, is isomerised to the corres-ponding 3-cephem compound and, if desired or required, the additional process steps are carried out.
The isomerisaticn of a 2-cephem compound to the corresponding 3-cephem compound can be carried out in a manner which is in itself known.
Thus, a 2-cephem compound of the formula VIII can be isomerised by treating it with a weakly basic agent and isolating the corresponding 3-cephem compound from an equi-librium mixture which may be obtained.
Suitable isomerisation agents are, for example, organic nitrogen-containing bases, such as tertiary hetero-cyclic bases of aromatic character, and above all tertiary aliphatic, azacycloaliphatic or araliphatic bases, such as N,N,N-tri-lower alkylamines, for example N,N,N-trimethylamine, N,N-dimethyl-N-ethylamine, N,N,N-triethylamine or N,N-diiso-propyl-N-ethylamine, N-lower alkyl-azacycloalkanes, for example N-methyl-piperidine, or N-phenyl-lower alkyl-N,N-di-lower alkyl-amines, for example N-benzyl-N,N-dimethylamine, as well as mixtures thereof, such as the mixture of a base of lVSS~4 the pyridine type, for example pyridine, and of a N,N,N-tri-lower alkylamine, for example pyridine and triethylamine.
It is also possible to use inorganic or organic salts of bases, especially of medium-strength to strong bases with weak acids, such as alkali metal salts or ammonium sal-ts of lower alkanecarboxylic acids, for example sodium acetate, triethyl-ammonium acetate or N-methyl-piperidine acetate, as well as other analogous bases or mixtures of such basic agents.
me above isomerisation with basic agents can be carried out, for example, in the presence of a derivative of a carboxylic acid which is suitable for forming a mixed anhydride, such as a carboxylic acid anhydride or halide, for example with pyridine in the presence of acetic anhydride.
The process is preferably carried out in an anhydrous medium, in the presence or absence of a solvent, such as an option-ally halogenated, for example chlorinated, aliphatic, cyclo-aliphatic or aromatic h~rdrocarbon, or of a solvent mixture, it being possible for bases which are used as reactants and are liquid under the reaction conditions at the same time also to serve as solvents, if necessary with cooling or heating, preferably in a tempera+vure range of about -30C to about +100C, in an inert gas atmosphere, for example a nitrogen atmosphere, and/or in a closed vessel.

3-Cephem compounds thus obtainable can be separated from 2-cephem compounds of the formula VIII which may still be present, in a manner which is in itself known, for example by adsorption and/or crystallisation.
m e isomerisation of 2~cephem compounds of the ¢.,~ ~1 ~5S5~
~ .
formula VIII can also be carried out by oxidising these in the l-posi-tion, if desired separating an isomer mixture, which may be obtained, of the l-oxides of corresponding 3-cephem compounds, and reducing the l-oxides of the corresponding 3-cephem compoun~, -thus obtainable.
Suitable oxidising agents for the oxidation of 2-cephem compounds in the l-position are inorganic per-acids which have a reduction potential of at least +1.5 volt and consist of non-metallic elements, organic per-acids or mix-tures of hydrogen peroxide and acids, especially organic car-boxylic acids, having a dissociation constant of at least 10 5. Suitable inorganic per-acids are periodi~ acid and persulphuric acid. Organic per-acids are corresponding per-carboxylic acids and persulphonic acids which can be added as such or can be formed in situ by using at least one equivalent -of hydrogen peroxide and of a carboxylic acid. It is desir-able to use a large excess of the carboxylic acid if, for example, acetic acid is used as the solvent. Examples of suitable per-acids are performic acid, peracetic acid, per-trifluoroacetic acid, permaleic acid, perbenzoic acid, mono-perphthalic acid or p-toluenepersulphonic acid.
The oxidation can also be carried out using hydrogen peroxide with ca-talytic amounts of an acid having a dissocia-tion constant of at least 10 5, it being possible to employ low concentra-tions, for example 1-2% and less, but also larger amounts, of the acid. The effectiveness of the mix-ture above all depends on the strength of the acid. Examples of suitable mix-tures are those of hydrogen peroxide wi-ch ~.. j .

1()559~4 acetic acid, perchloric acid or trifluoroacetic acid.
The above oxidation can be carried out in the presence of suitable catalysts. Thus, for example, the oxidation with percarboxylic acids can be catalysed by the presence of an acid having a dissociation constant of at least 10-5, its effectiveness depending on its strength. ~cids suitable for use as catalysts are, for example, acetic acid~ per-chloric acidand trifluoroacetic acid. Usually at least equimolar amounts of the oxidising agent, and preferably a slight excess of about 10% to about 20%, are used. The oxida-tion is carried out under mild conditions, for example at temperatures of about -50C to about +100C, preLerably of about -10C to about +40G.
The oxidation of 2-cephem compounds to -the l-oxides of the corresponding 3-cephem compounds can also be carried out by treatment with ozone, and also with organic hypohalite compounds, such as lower alkyl hypochlorites, for example tert.-butyl hypochlorite, which are used in the presence of inert solvents, such as optionally halogenated hydrocarbons, for example methylen~ cl;loride, and at temperatures of about -10C to about +30C, with periodate compounds, such as alkali metal periodates, for example potassium periodate, which are preferably used in an aqueous medium at a pH value of about 6 and at temperatures of about -10C to about +30C, with iodobenzene dichloride, which is used in an aqueous medium, preferably in the presence of an organic base, for example pyridine, and with cooling, for example at tempera-tures of about -20C to about 0, or with any other oxidising C~ 4~
_~ _ ll)S5~;~4 agent which is suitable for converting a thio grouping to a sulphoxide grouping.
In the l-oxides of 3-cephem compounds, thus obtain-able, it is possible, if desired, to carry out the additional process steps mentioned in connection with the isomerisation process. Further, a mixture of isomeric a- and ~-l-oxides can be separated, for example chromatographically.
The reduction of the l-oxides of 3-cephem compounds can be carried out in a manner which is in itself known, by treatment wi-th a reducing agent, if necessary in the presence of an activating agent. Possible reducing agents are:
catalytically activated hydrogen, in which case noble metal catalysts are used which contain palladium, platinum or rhodium and are optionally employed together with a suitable support, such as charcoal or barium sulphate; reducing tin, iron, copper or manganese cations which are used in the form of corresponding compounds or complexes of inorganic or organic nature, for example as tin-II chloride, fluoride, acetate or formate, iron-II chloride, sulphate, oxalate or succinate, copper-I chloride, benzoate or oxide or manganese II
chloride, sulphate, acetate or oxide, or as complexes, for example with ethylenediaminetetraacetic acid or nitrolctriacetic acid; reducing dithionite, iodide or ferrocyanide anions~
which are used in the form of corresponding inorganic or organic salts, such as alkali metal salts, for example sodium dithionite or potassium dithionite, sodium iodide or potassium iodide, or sodium ferrocyanide or potassium ferrocyanide, or in the form of the corresponding acids, such as hydriodic acid;

~'1 ~
~, l 1~559~4 reducing trivalent inorganic or organic phosphorus compounds, such as phosphines, and also esters, amides and halides of phosphinous, phosphonous or phosphorous acid, as well as phosphorus-sulphur compounds corresponding to -these phos-phorus-oxygen compounds, in which phosphorus compounds organic radicals above all represent aliphatic, aromatic or araliphatic radicals, for example optionally substituted lower alkyl, phenyl or phenyl-lower alkyl groups, such as, for example, triphenylphosphine, tri-n-butylphosphine, diphenyl-phosphinous acid methyl ester, diphenylchlorophosphine, phenyldichlorophosphine, benzenephosphono~s acid dimethyl ester, butanephosphonous acid methyl ester, phosphorous acid biphenyl ester, phosphorous acid trimethyl ester, phosphorus trichloride, phosphorus tribromide and the like; reducing halogenosilane compounds which possess at least one hydrogen atom bonded to the silicon atom and which in addition to halo-gen, such as chlorine, bromine or iodine, can also contain organic radicals, such as aliphatic or aromatic groups, for example optionally substituted lower alkyl or phenyl groups, such as chlorosilane, bromosilane, dichlorosilane or tri-chlorosilane, dibromosilane or tribromosilane, diphenylchloro-silane, dimethylchlorosilane and the like; reducing quater-nary chloromethylene-iminium salts, especially chlorides or bromides, wherein the iminium group is substituted by a bi-valent or two monovalent organic radicals, such as optionally substituted lower alkylene or lower alkyl groups, such as N- .
chloromethylene-N,N-diethyliminium chloride or N-chloromethyl-ene-pyrrolidinium chloride, and complex metal hydrides, such -i ~ /

l()S5~4 as sodium borohydride, in the presence of suitable activating agents, such as cobalt-II chloride, as well as borane dichloride.
Activating agents to be mentioned, which are used together with those of the abovementioned reducing agents which do not themselves possess Lewis acid properties, that is to say above all together with the dithionite, iodide or ferrocyanide reducing agents and the non-halogen-Gontaining trivalent phosphorus reducing agents or in -the catalytic reduction, are in particular, organic carboxylic acid halides and sulphonic ac.d halides and also sulphur, phosphorus or silicon halides wi.-th second order hydrolysis constants which are equal to or greater than that of benzoyl chloride, for example phosgene oxalyl chloride, acetic acid chloride or acetic acid bromide, chloroacetic acid chloride, pivalic acid chloride, 4-methox-ybenzoic acid chloride, 4-cyanobenzoic acid chloride, p-toluenesulphonic acid chloride, methanesulphonic acid chloride, thionyl chloride, phosphorus oxychloride, phosphorus trichloride, phosphorus tribromide, phenyldi-chlorophosphine, benzenephosphonous acid dichloride, dimethyl-chlorosilane or trichlorosilane and also suitable acid anhy-drides, such as trifluoroacetic anhydride, or cyclic sultones, such as ethanesultone, 1,3-propanesultone, 1,4-butanesultone or 1,3-hexanesultone.
The reduction is preferably carried out in the pre-sence of solvents or mixtures thereof, the choice of which is above all determined by the solubility of the starting mater-ials and the choice of the reducing agent, such as, for r ~
~ ~.

5~4 .
example, lower alkanecarboxylic acids or esters thereof, such as acetic acid and ethyl acetate, in the case of the cata-lytic reduction,and, for example9 optionally substituted, such as halogenated or nitrated, aliphatic, cycloaliphatic, aromatic or araliphatic hydrocarbons, for example benzene, methylene chloride, chloroform or nitromethane, suitable acid derivatives, such as lower alkanecarboxylic acid es-ters or nitriles, for example ethyl acetate or acetonitrile, or amides of inorganic or organic acids, for example dimethylformamide or hexamethylphosphoramide, ethers, for example diethyl ether, tetrahydrofurane or dioxane, ketones, for example acetone, or sulphones, especially aliphatic sulphones, for example dimethylsulphone or tetramethylenesulphone and the like, together with the chemical reducing agents, these sol-vents preferably not containing any water. The process is generally carried out at temperatures of about -20C to about 100C, and when using very reactive activators the reaction can be carried out at lower temperatures.
In a compound of the formula I oktainable according to the invention, which contains a free carboxyl group of the formula -C(=0)-R, and in which the amino of the aminomethyl radical is optionally present in a protected form, the free carboxyl group can be converted in a manner which is in itself known into an esterified carboxyl group w~ich can be split under physiological conditions. Thus, for example, in a compound of the formula I having a free carboxyl group~or in a salt thereof, for ex~ple in an alkali metal salt, such as a sodium salt or potassium salt, or an alkaline earth metal ~3 ., _,~
~.,, ~3S~Z~

salt, such as a calcium or magnesium salt, or in an optionally substituted ammonium salt, such as the triethylammonium salt, thereof, the carboxyl group can be converted into the corresponding esterified carboxyl group -C(=0)-R by reaction with a suitable halide, for example chloride or bromide.
Furthermore, it is possible, in a compound of the formula I obtain-able according to the invention, wherein the amino group in the aminomethyl substituent is preferably protected, to replace the group Rl in a manner which is in itself known by another radical Rl or convert it to another radical R
as defined above. Thus it is possible, for example, in a compound of the formula I, wherein Rl denotes a group of the formula -CH2-R2 and R2 represents, for example, a radical replaceable by nucleophilic substituents, or in a salt thereof, to replace such a radical R2, by treatment with a mercaptan compound, by an etherified mercapto group R2. A suitable radical replaceable by an etherified mercapto group is, for example an esterified hydroxyl group, for example a hydroxyl group esterified by aliphatic lower alkanoyloxy, as well as by a carbonic acid half-derivative as mentioned above. Such an esterified hydroxyl group is in particular acetoxy.
The reaction of such a compound with a suitable mercaptan compound can be carried out under neutral or weakly basic conditions, in the presence of water and optionally of a water-miscible organic solvent. The basic condi-tions can be obtained, for example, by adding an inorganic base, such as an alkali metal hydroxide, carbonate or bicarbonate or alkaline earth metal hy-droxide, carbonate or bicarbonate, for example sodium hydroxide, carbonate or bicarbonate, potassium hydroxide, carbonate or bicarbonate or calcium hydroxide, carbonate or bicarbonate. As organic solvents it is possible to use, for ex-ample, water-miscible alcohols, for example lower alkanols, such as methanol or ethanol, ketones, for example lower alkanones such as acetone, amiues, for example lower alkanecarboxylic acid amides, such as dimethylformamide, and the like.
Esterified hydroxyl groups R2 in a compound of the formula I, wherein Rl denotes the group -CH2-R2, wherein R2 represents a hydroxyl group esterifi-ed by the acyl radical of an optionally substituted half-amide of carbonic acid, ~, .
~ , -54-l~S59;~4 can be introduced, for example, by reacting a corresponding compound of the formula I, wherein R2 represents free hydroxyl, ~which can be liberated, for example, by removing the acetyl radical from an acetoxy group R2, for example by hydrolysis in a weakly basic medium, such as an aqueous sodium hydroxide solution at pH 9-10, or by treatment with a suitable esterase, such as an ap-propriate enzyme from Rhizobium tritolii, Rhizobium lupinii, Rhizobium japoni-cum or Bacillus subtilis, or a suitable citrus esterase, for example from orange peel), with a suitable carbonic acid derivative, especially with an isocyanate compound or carbamic acid compound, such as a silylisocyanate, for example silyltetraisocyanate, a sulphonylisocyanate, for example chlorosul-phonylisocyanate, or carbamic acid halide, for example carbamic acid chloride (which lead to N-unsubstituted 3-aminocarbonyloxymethyl compounds), or with a N-substituted isocyanate or a N-mono-substituted or N,N-disubstituted carbamic acid compound, such as a corresponding carbamic acid halide, for example carba-mic acid chloride, the reaction usually being carried out in the presence of a solvent or diluent and, if necessary, with cooling or warming, in a closed vessel and/or in an inert gas atmosphere, for example a nitrogen atmosphere.
Salts of compounds of the formula I can be prepared in a manner which is in itself known. Thus it is possible to form salts of compounds of the formula I which have acid groups, for example by treatment with metal compounds, such as alkali metal salts of suitable carboxylic acids, for example the sodi-um salt of ~-ethyl-caproic acid, or with ammonia or with a suitable organic amine, preferably using stoichiometric amounts or only a slight excess of the salt-forming agent. Acid addition salts of compounds of the ~ ; -55-lV~

formula I are obtained in the usual manner, for example by treatment with an acid or a suitable anion exchange reagent.
Inner salts of compounds of the formula I~which contain a free carboxyl group~can be formed, for example, by neutral-isation of salts, such as acid addition salts, to the iso-electric point, for example with weak bases, or by treatment with liquid ion exchangers.
Salts can be converted into the free compounds in the usual manner, metal salts and ammonium salts, for example, by treatment with suitable acids, and acid addition salts, for example, by treatment with a suitable basic agent.
The process also encompasses those embodiments accord-ing to which compounds arising 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; further, starting materials can be used in the form of derivatives or be formed during the reaction.
Preferably, those starting materials are used, and the reaction conditions are so chosen, that the compounds listed above as being par-ticularly preferred are obtained.
Starting materials of the formula II, wherein the amino group is optionally substituted by a group which permits acylation are known and can be manufactured according to known methods.
Thus, compounds of the formula II can be obtained when, in corresponding compounds of the formula sG
,~
,,, 1~)5~ 4 H2N =~ S \ ~jH2 \ C~ 1 (IX) C
Ro O

the amino group is converted into the diazo group, for example by treatment with a suitable diazotising agent, especially with nitrous acid or with nitro-gen tetroxide, the diazo compound is treated with a halogenoazide, for example bromoazide, and the 7-azido-7-halogeno-3-cephem compound, thus obtainable, wherein halogen above all represents bromine, and which can under certain circumstances be in the form of the 7-epimer mixture, is reacted with a suit-able silver salt, such as silver-I tetrafluoborate, in the presence of methan-ol. This gives the 7 ~azido-7~-methoxy-3-cephem compound in which the azido group is converted into the amino group by reduction, for example by catalytic hydrogenation in the presence of a noble metal catalyst, such as platinum oxide or palladium on charcoal and, if necessary, of an activating agent, such as a cobalt salt, for example cobalt-II acetate. This reaction sequence is describ-ed inter alia, by, for example, Cama et al,, J. Am. Chem. Soc., volume 94, page 1,408 (1972).
In a resulting starting material of the formula II the free amino group can be converted into a substituted amino group which permits acylation, for example by silylation or stannylation, such as by treatment with a suitably substituted silyl halide, for example trimethylsilyl chloride, lVSS~

or by treatment with an aldehyde, especially with an aryl-carboxaldehyde, for example an optionally substituted benz-aldehyde Starting materials of the formula III are known or can be manufactured according to methods which are in themselves known. Thus, for example, the 5-aminomethyl-2-thienyl-acetic acid or 5-aminomethyl-2-furyl-acetic acid compounds can be obtained.by acetylating an aminomethylthiophene or amino-methylfurane, wherein the amino group is preferably protected, for example by one of the protective groups mentioned, or by-acetylating an ac.d addition salt thereof, such as the hydro-chloride, by treatment with a suitable acetylating agent, for example with an anhydride of acetic acid 9 including an acetyl halide, such as acetyl chloride, or preferably with acetic anhydride, in -the presence of a suitable catalyst, such as a Lewis acid, for example aluminium chloride or aluminium bromide, or an acid, .such as polyphosphoric acid or preferably trifluoroacetic acid, or i-ts anhydride. The reaction can be carried out in an anhydrous solvent, such as an aromatic hydrocarbon,for examp~.'e benzene, or an excess of the liquid reagents employed, for example an excess of trifluoroacetic acid or acetic acid and/or their anhydrides. If trifluoro-acid or trifluoroacetic anhydride are used, a free amino group can at the same time be acylated by the trifluoroacetyl radi-cal.
An acetyla-ted aminomethylthiophene or aminomethyl-furane compound, wherein the amino group is preferably pro-tected, for example by the trifluoroacetyl radical, can be C., ' S~
_ ~ _ ~555~4 converted into a compound of the formula (III), for example by the method of Willgerodt or Willgerodt-Kindler, for example by heating with ammonium polysulphide or with a primary or secondary amine, such as morpholine, and subsequent hydro-lysis of the thioamide formed as an intermediate.
On the other hand, an acetylated aminomethylthiophene or aminomethylfurane compound, especially a compound of this type wherein the amino group is protected in the indicated manner, for example by the trifluoroacetyl radical, can be converted into a lower alkyl ester, for example the methyl ester, of an acid of the formula III by warming with thallium-(III) nitrate in the presence of a lower alkanol, especially methanol, and of an acid, for example perchloric acid, and the free acid can be prepared from the es~er by hydrolysis.
In the hydrolysis of a thioamide obtained by the Willgerodt or Willgerodt-Kindler method, or of an ester obtained by the thallium-(III) nitrate method, it is possible, depending on the hydrolysis conditions and the nature of the amino protective groups which may be present, also to split off these protective groups or, if desired, convert them.
The complete hydrolysis to a compound of the formula III and the subsequent introduction of an amino protective group can also be carried out in one step. For example, a resulting methyl ester of a compound of the formula III, wherein the amino group is acylated, for example with the trifluoroacetyl group, can first be hydrolysed by treatment with a base, for example an alkali metal hydroxide, such as sodium hydroxide, in water)or water together with a water-miscible organic -- ~iZ2 --~055~;~4 solvent, such as dioxane, and the product can then be treated in the same reaction mixture, for example with tert. butoxy-carbonyl azide, whereupon, after acidification and customary working up, the desired thiophene-acetic acid or furane-acetic acid having a tert.-butoxycarbonylaminomethyl group can be obtained.
Furthermore,it ispossible, for example, to obtain

4-aminomethyl-2-thienyl-acetic acid compounds or 4-amino-methyl-2-furyl-acetic acid compounds when 2-acetyl-thiophene or 2-acetyl-furane is halogenomethylated, especially chloro-methylated, for example by treatment with formaldehyde or a derivative thereof, such as paraformaldehyde, in the presence of a hydrogen halide acid, such as hydrochloric acid, and in the 2-acetyl-4-halogenomethyl-thiophene or 2-acetyl-4-halogenomethyl-furane compound thus obtainable, wherein halogen above all denotes chlorine, halogen is converted in a manner which is in itself known, for example by treatment with an alkali metal compound, for example potassium compound, of a phthalic acid imide, and subsequent hydrazinolysis, or by treatment with a suitable azide, such as an alkali metal azide or ammonium azide, into an optionally protected amino group and the acetyl substituent is then converted into the desired carboxymethyl radical, for example in accordance with the process described above.
In a compound of the formula III with an unprotected amino group, the latter can be converted into one of the pro-tected amino groups mentioned in accordance with any known method. Thus, an acyl radical can be introduced, as an Go __~ _ ~"~ ,~

ll)SS~ 4 amino protecti~e group, into the amino group, for example in accordance with the acylation process described above and also by treatment with a carbonic acid halide compound or carbonic acid azide compound, such as tert.-butoxycarbonyl-azide. Further, the free amino group can be substituted by a diarylmethyl or triarylmethyl group, for example by treat-ment with a reactive ester of a diarylmethanol or triaryl-methanol, such as trityl chloride, preferably in the presence of a basic agen-t, such as pyridine.
An amino group can also be protected by introducing a silyl group and stannyl group. Such groups are intro-duced in a manner which is in itself kno~n, for example by treatment with a suitable silylating agent, such as a dihalogeno-di-lower alkylsilane or tri-lower alkylsilyl halide, for example dichloro-dimethylsilane or trimethyl-silyl chloride, or an optionally N-mono-lower alkylated, N,N-di-lower alkylated, N-tri-lower alkyl-silylat~d or N-lower alkyl-N-tri-lower alkyl-silylated N-(tri-lower alkyl-silyl)-amine (see, for example, British Patent No. 1,073,530), or with a suitable stannylating agent, such as a bis-(tri-lower alkyl-tin) oxide, for example bis-(tri-n-butyl-tin) oxide, a tri-lower alkyl-tin hydroxide, for example triethyl-tin hydrox-de, a tri-lower alkyl-lower alkoxy-tin compound, tetra-lower alkoxy-tin compound or tetra-lower alkyl-tin compound as well as a tri-lower alkyl-tin halide, for example tri-n-butyl-tin chloride (see, for example, Netherlands Published Specification 67/11,107).
An amino group can also be protected b-y introduction , ~{)559~4 of a 2-carbonyl-1-vinyl group, whereby enamine or ketimine compounds are formed. Such groups can be obtained, for example, by treating the amine with a 1,3-dicarbonyl compound, for example with acetoacetic acid methyl ester or acetoacetic acid N,N-dimethylamide, in an anhydrous medium, for example a lower alkanol, such as methanol.
Arylthio or aryl-lower alkyl-thio protective groups, and also arylsulphonyl protective groups, can be introduced into an amino group by treatment with a corresponding aryl--thio or aryl-lower alkylthio and also arylsulphonyl halide, for example chloride.
The reactive functional acid derivatives of an acid of the formula III can be prepared in a manner which is in itself known. Acid halides are obtained, for example, by reacting a compound of the formula III, optionally having a protected amino group, or a salt thereof, with a halogen-ating agent, for example with an acid halide, sucn as an acid fluoride or acid chloride, of an inorganic acid containing phosphorus or sulphur, for example phosph~rus pentachloride, thionyl chloride or oxalyl chloride. The reaction is pre-ferably carried out in a non-aqueous solvent or solvent mix-ture, such as a carboxylic acld amide, for example dimethyl-formamide. The resulting acid halide does not have to be purified further but can be reacted direct with tne starting material of the formula II, using, for example, the same sol-vents or solvent mixtures which are used when preparing the acid halide.
Symme-trical anhydrides or mixed anhydrides, other ~ss9;~

than halides, of compounds of the formula III having an optionally protected amino group can be prepared, for example, by reacting a corresponding compound having a free carboxyl group, preferably a salt, especially an alkali metal salt, for example a sodium salt, or an ~onium salt, for example a tri-ethylammonium salt, thereof, with a reactive deriva-tive, such as a halide, for example the chloride, of a suitable acid, for example a halogenoformic acid iower alkyl ester, for example chloroformic acid isobutyl ester, or a lower alkanecarboxylic acid halide, for example trichloroacetic acid chloride.
Acti-vated esters of compounds of the formula III
having an optionally protected amino group can be prepared, for example, by reacting a corresponding compound having a free carboxyl group with an optionally substituted, for example nitro-substituted or hydrogen-substituted, such as chlorine-substituted, phenol, such as a nitrophenol, for example 4-nitrophenol or 2,4-dinitrophenol, or a polyhalogeno-phenol, for example 2,3,4,5,6-pentachlorophenol, in the presence of a carbodiimide, for example N,N'-dicyclohexyl-carbodiimide.
The starting materials of the formula IV can be manu-factured by introducing the group of the formula ~l2I~ l2 ~ 2- ~~ . (IIIa) wherein amino of the aminomethyl group is prefe~ably in a protected form, into the amino group of compounds of the ~3 ~ _~ _ ... ., ~

~ ~S ~

formula IX, wherein the amino group can optionally be substituted by a group which permits acylation, above all by acylation, for example in accordance with the process, described above, for the acylation of starting materials of the formula II.
The starting materials of the formula IVb can be obtained, for ex-ample, by a method wherein a 7 ~amino-3-cephem compound of the formula IX, wherein functional groups which are present, such as, for example, a carboxyl group of the formula -C(=0)-R, are preferably in a protected form, is used as the startlng point and converted, by reaction with an aldehyde, especially an aromatic aldehyde, such as benzaldehyde, into the Schiff's base, and the latter is reacted with an anion-forming agent, such as a preferably sterically hindered alkali metal lower alkanoate, such as potassium tert.-butylate, an alkali metal hydride, for example sodium hydride, an alkali metal-hydrocarbon compound, for example n-butyl-lithium or phenyl~lithium, or a suitable alkali metal compound of a secondary organic base, such as, for example, the lithium compound of a di-lower alkylamine or lower alkyleneamine, such as lithium di-ethylamide, preferably with cooling, for example at temperatures of about -30C to about 0C, and in the presence of a solvent or diluent, such as glycol dimethyl ether. The etherified mercapto group of the formula R-S- can be introduced into the anion thus obtainable either directly, for example by treatment with a suitable thiolsulphonic acid ester, such as a lower alkyl-thiolsulphonic acid lower alkyl esterp for example methanethiolsulphonic acid methyl ester, or with a sulphenyl halide, such as a lower alkylsulphenyl hal-ide, for example methylsulphenyl chloride, or indirectly via the corresponding 7~-fluoro-3-cephem-Schiff's base; the latter is obtained, for example, by treatment with ~luorine perchlorate and can be converted to the desired 7~-R-thio-3-cephem-Schiff's base by reaction with a mercaptan, such as a lower alkyl-mercaptan, for example methylmercaptan, in the presence of a strong acid, such as an optionally halogenated lower alkene-carboxylic acid, for example trifluoroacetic acid. In such an intermediate product, the nitrogen atom of the methylene-amino group is acylated by introduction of the group of the for-mula IIIa, wherein amino of the aminomethyl group is preferably present in a ~ ~, ~ ~64-protected form, for example in accordance with the process described above, thus giving the starting material of the formula IVb. This reaction sequence is carried out, for example, according to the methods described by Slusarchyk et al., J. Org. Chem., volume 38, page 943 (1973) and Spitzer and Goodson, Tetrahedron Letters, page 273 (1973).
The starting material of the formula V can be obtained, for example, by a method wherein, in a 3-cephem compound of the formula HN r ~ s ~ IH2 ¦ N C-Rl (X) 0=~
~CH2)3 ,,C ~
~
H N CH C
\ Ro wherein the carboxyl group -C ~ is usually in a protected form, the amino Ro group in the 5-amino-5-carboxy-valeryl radical is converted into a protected amino group AmO, and in a compound thus obtainable the acyl radical of the formula IIIa, wherein amino of the aminomethyl group is present in a protected form which differs in the method by which it can be split off, from that of the protected amino group AmO, is introduced into the nitrogen atom of the amide grouping, for example by acylation in accordance with the process des-cribed above, for example by treatment with an acid halide, for example acid chloride, of a compound of the formula III and a suitable silylating agent, such as a monosilylated or disilylated acid amide, such as an optionally halo-genated N-mono-tri-lower alkylsilyl- or N,N-bis-tri-lower alkylsilyl-lower alkanecarboxylic acid amide, which latter can also be in the N,O-bis-tri-lower alkylsilylated enol form of the amide, for example bis-trimethylsilyl-acetic acid amide or N-trimethylsilyl-trifluoroacetic acid amide, in a suitable sol-vent or diluent, for example in a halogenated hydrocarbon, such as methylene chloride, and, if necessary, with warming, in a closed vessel and/or in an ,~ ~
~ -65-~55~ 4 inert gas atmosphere, such as .

~t~5~

a nitrogen atmosphere. The preparation of the s-tarting material of the formula V in accordance with the above pro cess can be carried out, for example, in accordance with the method described by Sletzinger et al , J. Am. Chem. Soc., volume 94, page 1,410 (1972).
m e starting materials of the formula VI are known or can be manufactured in a manner which is in itself known, for example by acylation of the amino group in a compound of the formula II, for example by treatment wi-th an acid of the formula ~ CH2-C-OH (XI) or a suitable derivative, such as a mixed anhydride, especi-ally a halide, for example a chloride, thereof, for example in accordance with the acylation process described above.
The 2-cephem starting materials of the formula VIII
can be manufactured analogously to the process described above for the manufacture of the corresponding 3-cephem compounds, for example by acylation of the primary amino group in a com-pound of the formula 1 (XII) O==C-R

by treatment with an acid of the formula III, wherein amino bC I -,8~ _ ~V5~ 4 of the aminomethyl group is preferably in a protected form, or a reactive functional acid derivative thereof,or a sal-t of such a compound. The above acylation reac-tion can, for example, be carried out analogously to the methods described above.
In the process according to the invention, and in additional measures which may have to be carried out, and also in the manufacture of the star-ting materials it is possible, if necessary, temporarily to protect free functional groups, which do not participate in the reaction, in the starting materials or in the compounds obtainable in accordance with the process, as described above, for example free amino groups by acylation, tritylation or silylation, free hydroxyl groups or mercapto groups, for example, by etherification or esteri-fication, and free carboxyl groups, for example, by esteri-fication, including silylation, in a manner which is in itself known, and in each case, after the reaction has taken place, liberating the groups, if desired, in a manner which is in itself known, by solvolysis or reduction.
The pharmacologically usable compounds of the present invention can be used, for example, for the manufacture of pharmaceutical preparations which contain an ac-tive amount of the active substance together, or mixed, with inorganic or organic, solid or liquid, pharmaceutically usable excipients which are preferentially suitable for parenteral administration.
Preferably, the pharmacologically active compounds of the present invention are used in the form of injectable~
, --~ _ l()SS~4 for example intravenously administrable, preparations or of infusion solutions. Such solutions are preferably isotonic aqueous solutions or suspensions which can, for example, be prepared befo~e use from lyophilised preparations which contain the active substance by itself or together with an excipient, for example mannitol. The pharmaceutical preparations can be sterilised and/or contain auxiliaries, for example pre-servatives, stabilisers, wetting agents and/or emulsifiers, solubilising agents, salts for regulating the osmotic pressure and/or buffers. The present pharmaceutical preparations which, if desired, can contain further pharmacologically valu-able materials, are prepared in a manner which is in itself known, for example by means of conventional solution of lyo-philisation processes, and contain from about 0.1% to 100%, in particular from about 1% to about 50%, of lyophilisates with up to 100% of the active substance. Depending on the nature of the infection and the condition of the infected organism, daily doses of about 0.5 g to about 5 g administered subcutaneously are used for the treatment of warm-blooded animals weighing about 70 kg.
Unless defined otherwise, the term "lower" used in conjunction with the definition of organic radicals or com-pounds, for example in lower alkyl, lower alkanol and the like, denotes that the radicals or compounds in question have up to 7, preferably up to 4, carbon atoms.
The examples which follow serve to illustrate the invention.

~ ; ~~~
_ ,., _.

1~)559~4 Exam~le 1:
2.40 g of 3-acetoxymethyl-7~-[2-(5-tert.-butoxy-carbonylaminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylic acid diphenylmethyl ester are dissolved in 180 ml of tetrahydrofurane, the solution is cooled to between -70 and -75C under a nitrogen atmosphere and a solution of 0.46 g of lithium methoxide in 10 ml of methanol is added over the course of 1 minute, whilst stirring. After 3 minutes, 0.42 ml of tert.-butyl hypochlorite is added and the solution is stirred for a further 20 minutes at -70 to -75C, neu-tral-ised with 0.80 ml of acetic acid and concentrated to about 70 ml under a waterpump vacuum. 200 ml of water are added and the mixture is extracted with twice 300 ml of ethyl acetate. The organic extracts are washed wi-th water and with a saturated aqueous sodium chloride solution, dried over magnesium sulphate and evaporated under reduced pressure.
m e crude product is chromatographed on 250 g of silica gel, and 3-acetoxymethyl-7~-methoxy-7~-[2-(5--tert.-butoxycarbonyl-aminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylic acid diphenylme-thyl ester is eluted with a 6:4 mixture of toluene and ethyl acetate; thin layer chromatogram (silica gel):
Rf = 0.20 (system: toluene/ethyl acetate 6:4); ultraviolet absorption spectrum (in 95% strength aqueous ethanol):
~max = 243 m~ ( = 13,900) and ~min = 236 m~ ( ~ = 13,600);
infrared absorption spectrum (in methylene chloride): charac-teristic bands at 2.92 ~, 5.61 ~, 5.74 ~, 5.79 ~, 6.23 ~ and 6.67 ~.
The starting material can be prepared as follows:
~o _ ~ _ 10S~9'~4 ~ .
A solution of 20 g of 2-thenylamine hydrochloride in a mixture of 100 ml of -trifluoroacetic acid and 100 ml of acetic anhydride is stirred for 2 hours at 55C, with exclu-sion of moisture. The reaction mixture is concentrated com-pletely under reduced pressure, 50 ml of toluene are added and the mixture is again concentrated. The crude product is dissolved in ethyl acetate and the solution is treated-with active charcoal, filtered through silica gel and concen-trated under reduced pressure. After recrystallisation from diethyl ether, 2-acetyl-5-trifluoroacetylaminomethyl-thiophene is obtained, melting point 83-84C.
40 ml of 70% strength aqueous perchloric acid are added to a solution of 34.2 g of thallium-~) nitrate trihydrate in 100 ml of methanol whilst cooling with ice, and a solution of 20 g of 2-acetyl-5-trifluoroacetylamino-thiophene in 500 ml of methanol ls added dropwise over the course of 15 minutes at +5C under a nitrogen atmosphere.
The solution is warmed to 50C and stirred at -this temperature for 2~ hours. The reaction mixture is cooled to about +5 C
and poured into an ice-cold solution of 120 g of dipotassium hydrogen phosphate in 300 ml of water. The mixture is fil-tered and the filter residue is washed with methanol. The filtrate is concentrated to about 300 ml under reduced pres-sure and is extracted with three times 150 ml of chloroform.
The extracts are washed with water and subsequently with saturated aqueous sodium chloride solution, dried over mag-nesium sulphate and concentrated under a waterpump vacuum.
This leaves 2-(5-trifluoroacetylaminomethyl-2-thienyl)-~/
~

~:)S5~4 acetic acid me-thyl ester; thin layer chromatogram (silica gel: system: toluene/ethyl acetate, 60:40): Rf = 0.65;
infrared absorption spectrum (in methylene chloride):
characteristic bands at 2.93 ~, 3.39 ~, 5.75 ~ and 5.80 ~.
40 ml of 2 N aqueous sodium hydroxide solution are added to a solution of 9.7 g of 2-(5-trifluoroacetylamino-methyl-2-thienyl)-acetic acid methyl ester in 50 ml of dioxane at 20C under a nitrogen atmosphere. The mixture is stirred for 2 hours at 20-25C and diluted with 50 ml of dioxane, and 8.5 ml of tert.-butoxycarbonyl azide are added to the solution, after which it is stirred for a further 1~ hours at 20-25C.
The reaction mixture is cooled to about 5C and adjusted to pH 2.5 with about 40 ml of 20% strength aqueous phosphoric acid. It is concentrated to about 50 ml under reduced pressure and extracted with three times 200 ml of ethyl acetate. The extracts are washed with a saturated aqueous sodium chloride solution~ dried over magnesium sulphate, decolorised by treatment with an active charcoal preparation and concentrated under reduced pressure. The residue is recrystallised from diethyl ether, and 2-(5-tert.-butoxy-carbonylaminomethyl-2-thienyl)-acetic acid, melting poin-t 114-115C, is -thus obtained.
0.90 g of 2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetic acid is dissolved in 20 ml of methylene chloride (distilled over phosphorus pentoxide), containing 0.334 g of N-methylmorpholine, the solution, from which mois-ture is kept excluded, .7 S cooled -to -20C and 0.45 ml of chloroformic acid isobutyl ester is added dropwise, whilst , j -- ~ _ ~VSS~;~4 keeping the temperature at between -15C and -20C. After 30 minutes, a solution of 1.04 g of 3-acetoxyme-thyl-7~-amino-3-cephem-4-carboxylic acid diphenylmethyl ester in 5 ml of methylene chloride is added, after which stirring is contin-ued for 2 hours at -10C and for 8 hours at room tempera-ture.
The mixture is poured into ice-cold water, adjusted to pH
8.0 with dipotassi~ hydrogen phosphate and repeatedly extrac-ted with methylene chloride. me organic extracts are washed with a saturated aqueous sodium chloride solution and dried over magnesium sulphate, after which the solvent is removed under reduced pressure. This g~ves 3-acetoxymethyl-7~-[2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetyl-amino]-3-cephem-4-carboxylic acid diphenylmethyl ester as a colourless foam which is directly converted further; thin layer chromatogram (silica gel): Rf = 0.71 (system: hexane/
ethyl acetate/methanol, 20:40:40). The product can be crystallised from diethyl ether, melting point 134-136C;
infrared absorption spectrum (in mineral oil): character-istic bands at 3.03 ~, 5.68 ~, 5.77 ~, 6.02 ~, 6.26 ~ and 6.57 ~; ultraviolet absorption spectrum (in 95% strength aqueous ethanol): ~max = 245 m~ (~ = 15,100).
The starting material can also be obtained as follows:
1.42 g of 2-thienylacetic acid are added in portions to a solution, kept at 0-5C, of 1.34 g of N-hydroxymethyl ace-tamide (Einhorn, Ann. Chem., volume 343, page 264 (1905)) in 10 ml of trifluoroacetic acid, whilst stirring. The mixture is stirred for 2 hours at 0-5C and the trifluoro-acetic acid is distilled off under reduced pressure.

p.~ 73 - _&~ -l~5S9;~4 30 ml of water are added to the residue and the mixture is extracted with twice 50 ml of ethyl acetate. The organic extracts are washed with a saturated aqueous sodium chloride solution, dried over magnesium sulphate, concen-trated under reduced pressure and filtered through silica gel. m e residue is crystallised from ethyl acetate and gives 2-(5-acetylaminomethyl-2-thienyl)-acetic acid, thin layer chromato-gram (silica gel): Rf = 0.61 (system: butanol/acetic acid/
water, 45:45:10); infrared absorption spectrum (mineral oil):
characteristic bands at 2.95 ~, 5.82 ~1, 6.23 ~-and 6.38 ~.
0.3 ml of a 2 ~ aqueous sodium hycroxide solution is added to a solution of 0.6 g of 2-(5-acetylaminomethyl-2-thienyl)-acetic acid in 5 ml of dioxane. me mixture is stirred for 12 hours at 55-60C and cooled to 25C, 0.2 ml of tert.-bu-toxycarbonyl azide is added, the mixture is stirred for 16 hours at room temperature and 2-(5-tert.-butoxycar-bonylaminomethyl-2-thienyl)-acetic acid is isolated in accord-ance with the process described above.
Example 2:
A solution of 0.247 g of 3-acetoxymethyl-7~-azido-7a-methoxy-3-cephem-4-carboxylic acid diphenylmethyl ester (Cama et al., J. Am. Chem. Soc. volume 94, page 1,408 (1972);
German Offenlegungsschrift 2,129,675) in 7.6 ml of tetra-hydrofurane is hydrogenated in the presence of 0.167 g of platinum oxide and 0.076 g of cobalt-II acetate at room temperature for 1~ hours with hydrogen under a pressure of 2 atmospheres. m e reaction mixture is filtered and the filtrate, containing the 3-acetoxymethyl-7~-amino-7a-methoxy--?Y
-- 8~ --` ` 105~9~4 3-cephem-4-carboxylic acid diphenylmethyl ester is used without further purification in the subsequent acylation step.
A solution of 0.237 g of 2-(5-tert.-butoxycarbonyl-aminomethyl-2-thienyl)-ace-tic acid in 20 ml of methylene chloride is treated at -15C with 0.095 ml of 4-methylmor-pholine, followed by 0.119 ml of chloroformic acid isobutyl ester. The mixture is stirred for 15 minutes at -15C and the above solution of the 3-acetoxy-methyl-7~-amino-7a-methoxy-3-cephem-4-carboxylic acid diphenylmethyl es-ter is then added dropwise at the same temperature. me reaction mixture is stirred at 0C for 3 hours 3 then diluted with methylene chloride and washed with a saturated aqueous solu-tion of sodium bicarbonate and a saturated aqueous solution of sodium chloride. The aqueous wash liquids are extracted with methylene chloride and the combined organic extracts are dried over sodium sulphate, filtered and evaporated under reduced pressure. The residue is subjected to a thick layer chromatogram (silica gel), which is developed with a

5:3 mixtu-e of toluene and ethyl acetate. 3-Acetoxymethyl-7~-C2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetyl-amino]-7a-methoxy-3-cephem-4-carboxylic acid diphenylmethyl ester of Rf = 0.2 is obtained, and is identical with the pro-duct of the process described in Example 1.
Example 3:
0.168 g of chloromethylene-dimethyl-ammonium chloride (obtained as a solid substance by reaction of equimolar amounts of dimethylformamide and phosgene in methylene 7~' lV55~'~4 chloride) is added to a solution of 0.271 g of 2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetic acid in 5 ml of methylene chloride at 0C and the reaction mixture is stirred for lO minutes. 0.081 ml of pyridine is then added at 0C, the mixture is s'~irred for 5 minutes a-t this temperature and the solution prepared according to the process of Example 2, containing 3-acetoxymethyl-7~-amino-7a-methoxy-3-cephem-4-carboxylic acid diphenylmethyl ester, is added. The mixture is stirred for 30 minutes at 0C, then diluted with methylene chloride, and washed with a saturated aqueous solution of sodium bicarbonate and a saturated aqueous solution of sodium chloride. The aqueous wash liquids are extracted with methylene chloride and the combined organic extracts are dried over sodium sulphate, filtered and evaporated under reduced pressure. The residue is subjected to a thick layer chro-matogram (silica gel), which is developed with a 5:3 mixture of toluene and ethyl acetate. 3-Acetoxymet,hyl~7~-[2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetylamino]-7-methoxy-3-cephem-4-carboxylic acid diphenylmethyl ester of Rf = 0.2 is obtained and is identical with the p-oduct of the process described in Example 1.
Example 4:
A solution of 0.90 g of 3-acetoxyme-thyl-7~-me-thoxy-7~-[2-(5-tert~-butoxycarbonylaminomethyl-2-thienyl)-acetyl-amino]-3-cephem-4-carboxylic acid diphenylmethyl ester in 5 ml of trifluoroacetic aoid and 1.5 ml of anisole is left ~o stand for 30 minutes at 0C and evaporated, with addition of 20 ml of toluene, under a waterpump vacuum. The residue ~6 _ _~

- 11)55;9;~
is triturated with ]0 ml of die-thyl ether and 40 ml of petrol-eum ether and fil-tered. The filter residue, which contains the trifluoroacetic acid salt of 3-acetoxymethyl-7~-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-cephem-4-carboxylic acid is dried for 5 hours in a high vacuum at room temperature and then dissolved in 30 ml of methanol; the solution is decolorised by adding active charcoal, and filtered. The filtrate is adjusted to pH 6.0 with tri-ethylamine and is then left to stand for 2 hours at 0-5C.
The inner salt of 3-acetoxymethyl-7~-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-cephem^-4-carboxylic acid, which has precipitated, is filtered off and dried ~or 20 hours under a high vacuum at room temperature; melting point above 180C (with decomposition); [a]D20 = ~ 177 + 1 (c = 0.972 in an 0.15 molar aqueous potassium dihydrogen phosphate/
dipotassium hydrogen phosphate buf~er solution, pH 7.3 - 7.4);
thin layer chromatogram (silica gel): Rf = 0~12 (system:
n-butanol/acetic acid/water, 45:45:10); ultraviole-t absorp-tion spectrum (in an 0.15 molar aqueous potassium dihydrogen phosphate/dipotassium hydrogen phosphate buffer solution~ pH
7.3 ~ 7 4) ~max = 241 m~ ( ~ = 15,700)and ~min = 214 m~
( = 8,700); infrared absorption spectrum (in mineral oil):
characteristic bands at 5.66 ~, 5.75 ~, 6.00 ~, 6.34 ~ and

6.55 ~-Example 5:
0.028 g of sodium bicarbonate is added to a solutionof 0.15 g of the inner salt of 3-acetoxymethyl-7~-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7-methoxy-3-cephem-4-.~ _ ,~

~ . .

. 1055S~'~4 .
carboxylic acid and 0.053 g of the sodium salt of 5-mercapto-l-methyl-tetrazole in 7 ml of acetone and 10 ml of water, and the mixture is warmed to 60C for 5 hours. It is filtered and the filtrate is concentrated under reduced pressure to a volume of about 10 ml, adjusted to pH 5.5 with acetic acid and left to stand for some hours at 0-5C. The precipitate is filtered off and washed with acetone. This gives the inner salt of 7~-[2-(5-aminomethyl-2-thienyl)-acetylamino]-

7~-methoxy-3-(1-methyl-5-tetrazolyl-thiomethyl)-3-cephem-4-carboxylic acid, which is dried for 12 hours under a high vacuum; thin layer chroma-togram (silica gel): Rf = 0.25 (system: chloroform/methanol, 1:1).
In another variant, the reaction is carried out in the presence of 0.028 g of sodium bicarbonate and 0.055 g of potas-sium iodide.
Example 6:
9.1 g of 3-acetoxymethyl-7~-[2-(5-tert.-butoxycarbon-ylaminomethyl-2-furyl)-acetylamino]-3-cephem-L~-carboxylic acid diphenylmethyl ester are dissolved in L~oo ml of tetrahydro-~urane, the solution is cooled to between -70 and -75C under a nitrogen atmosphere and a solution of 1.80 g of lithium methoxide in 50 ml of methanol is added over the course of 1 minute, whilst stirring. After 2 minutes, 1.62 ml of tert.-butyl hypochlorite are added and the mixture is stirred for a further 20 minutes at -70 to -75C, neutralised with 2.5 ml of acetic acid and concentrated to about 100 ml under a water-pump~vacuum. 500 ml of water are added and the mixture is extracted with twice 500 ml of ethyl acetate. The organic _ ~[ _ ~055~;~4 , .
extracts are washed with water and with a saturated aqueous sodium chloride solution, dried over magnesium and evaporated under reduced pressure. The crude product is chromato-graphed on 600 g of silica gel, and 3-ace-toxymethyl-7~-methoxy-7~-[2-(5-tert.-butoxycarbonylaminomethyl-2-furyl)-acetylamino]-3-cephem-4-carboxylic acid diphenylmethyl ester is eluted with a 6:4 mixture of toluene and ethyl aceta-te; thin layer chromatogram (silica gel): Rf = 0.30 (system: toluene/ethyl acetate, 6:4); infrared absorption spectrum (in methylene chloride): characteristic bands at 2.92 ~, 5.61 ~, 5.76 !
(broad), 6.24 ~ and 6.67 ~.
The starting material can be prepared as follows:
A mixture of 50 g of furfurylamine and 150 ml of tri-fluoroacetic anhydride is stirred for 2 hours at room tempera-ture, with exclusion of moisture. 150 ml of acetic acid are added and the mixture is stirred for a further 2 hours at 55C. The reaction mixture is completely concentrated under reduced pressure, mixed with 50 ml of toluene and again con-centrated. The crude product is dissolved in ethyl acetate and the solution is treated with active charcoal, filtered through silica gel and concentrated under reduced pressure.
After recrystallisation from diethyl ether, 2-acetyl-5-tri-fluoroacetaminomethyl-furane, melting point 99-100C, is obtained; ultra~iolet aksorption spectrum (in ethanol):
Amax = 275 m~ ( æ = 15,500).
50 ml of 70% strength perchloric acid are added to a solution of 39.8 g of thallium-(III) nitrate trihydrate in 100 ml of methanol whilst cooling with ice and a solution of J~ ' _ .~..1 55~4 20 g of 2-acetyl-5-trifluoroacetaminomethyl-furane in 500 ml of methanol is added dropwise over the course of 15 minutes at +5C under a nitrogen atmospher~ The solution is warmed to 50C and stirred at this temperature for 22 hours. The reaction mixture is cooled to about +5C and poured into an ice-cold solution of 120 g of dipotassium hydrogen phosphate in 300 ml of water. The mixture is filtered and the filter residue is washed with methanol. The filtrate is concen-trated to about 300 ml under reduced pressure and extracted with three times 150 ml of chloroform. The extracts are washed with water and subsequently with a saturated aqueous sodium chloride solution, dried over magnesium sulphate and concentrated under a waterpump vacuum. This leaves 2-(5-trifluoroacetaminomethyl-2-furyl)-acetic acid methyl ester, thin layer chromatogram (silica gel): Rf = 0.35 (system:
toluene/ethyl acetate, 60:40); infrared absorption spectrum (in methylene chloride): characteristic bands at 2.49 ~, 3.39 ~ and 5.~0 ~.
- 75 ml of a 2 N aqueous sodium hydroxide solution are added to a solution cf 1~.4 g of 2-(5-trifluoroacetamino-methyl-2-furyl)-acetic acid methyl ester in 100 ml of dioxane at 20C under a nitrogen atmosphere. The mixture is stirred for 4 hours at 20-25C and diluted with 100 ml of dioxane, and 34 ml of tert.-butoxycarbonyl azide are added to the solution, after which the mixture is stirred for a further 16 hours at 20-25C. The reaction mixture is cooled to about 5C and adjusted to pH 2.5 with about 40 ml of 20%
strength aqueous phosphoric acid. The mixture is ~0 _~ _ ~; '` .

l()SS~4 , .
concentrated to a volume of about 100 ml under reduced pressure and is extracted with three times 200 ml of ethyl acetate. The extracts are washed with a saturated aqueous sodium chloride solution, dried over magnesi~m sulphate, decolorised by treatment with an active charcoal preparation, and concentrated. After recrystallisation from diethyl ether, 2-(5-tert.-butoxycarbonylaminomethyl-2-furyl)-acetic acid, melting point 72-73C, is obtained.
1.87 g of 2-(5-tert.-butoxycarbonylaminomethyl-2-furyl)-acetic acid are dissolved in 200 ml of methylene chloride (distilled over phosphorus pentox:ide) and 0.80 ml of 4-methylmorpholine, the solution, from which moisture is kept excluded, is cooled to -20C and 1.0 ml of chloroformic acid isobutyl ester is added dropwise whilst keeping the temperature at between -15C and -20C. After 30 minutes, a solution of 2.31 g of 3-acetoxyme-thyl-7~-amino-3-cephem-4-carboxylic acid diphenylmethyl ester is added, after which the mixture is stirred for a further 2 hours at -10C and for

8 hours at room temperature. It is then poured out into ice-cold water, the pH is adjusted to 8.0 with dipotassium hydro-gen phosphate and the mixture is repeatedly extracted with methylene chloride. The organic extracts are washed with a saturated aqueous sodium chloride solution and dried over magnesium sulphate, after which the solvent is removed under reduced pressure. 3-Acetoxymethyl-7~-[2-(5-tert.-butoxy-carbonylaminomethyl-2-furyl)-acetylamino]-3-cephem-4-carboxy-lic acid diphenylmethyl ester is obtained as a colourless foam which is directly conver-ted further; thin layer C~

105~9;~4 chromatogram (silica gel): Rf = 0.84 (system: hexane/ethyl acetate/methanol 7 20:40:40).
Example 7:
A solution of 2.95 g of 3-acetoxymethyl-7a-me-thoxy-7~-[2-(5-tert.-butoxycarbonylaminomethyl-2-furyl)-acetyl amino]-3-cephem~4~carboxylic acid diphenylmethyl ester in 3 ml of anisole and 12 ml of trifluoroacetic acid is left to stand for 30 minutes at 0C and is then evaporated, with addition of 100 ml of toluene, under a waterpump v~cuum.
me residue is triturated with 50 ml of diethyl ether and filtered. The filter residue, which contains the trifluoro-acetic acid salt of 3-acetoxymethyl-7~-[2-(5-aminomethyl-2-furyl)-acetylamino]-7a-methoxy-3-cephem-4-carboxylic acid, is dried for 5 hours under a high vacu~m at room temperature and then dissolved in 30 ml of methanol; the solution is decolorised by adding active charcoal, and filtered. The filtrate is adjus-ted to pH 6.o with triethylamine and then left to stand for 2 hours at 0-5C. The inner salt of 3-acetoxymethyl-7~-~2-(5-aminomethyl-2-furyl)-acetylamino]-7a-methoxy-3-cephem-4-carboxylic acid, which has precipi-tated, is filtered off and dried for 20 hours under a high vacuum at room temperature; melting point above 165C (with decom-position); thin layer chromatogram (silica gel): Rf = 0.22 (system: chloroform/methanol, 1:1); ultraviolet absorption spectrum (in 95% strength aqueous ethanol): ~max = 222 m~
~ max2 = 266 m~ (~ = 6,200); infrared absorption spectrum (in mineral oil): characteris-tic bands at 5.66 ~, 5.77 ~, 5.91 ~, 6.22 ~ and 6.52 ~.

l~)SS3;~4 Example 8:
4.0 g of 3-acetoxymethyl-7~-[2-(5-tert.-butoxycar-bonylaminomethyl 2-thienyl)-acetylamino]-3-cephem-4-carboxy-lic acid tert.-butyl ester are dissolved in 250 ml of tetra-hydrofurane, the solution is cooled to between -70C and -75C under a nitrogen atmosphere and a lithium methylate solution prepared by dissolving 0.170 g of lithium wire in 30 ml of methanol is added over the course of one minute, whilst stirring. After -three minutes, 0.80 ml cf ter-t.-butyl hypochlorite is added and the mixture is stirred for a fur-ther 20 minutes at -70C to -75C, neutra ised with 4.0 ml of acetic acid and concentrated to a volume of about 70 ml under a waterpump vacuum. 200 ml of water are added and the mix-ture is extracted with twice 300 ml of ethyl acetate. me organic extracts are washed with water and with a sa-turated aqueous sodium chloride solution, dried over magnesium and evaporated under reduced pressure. The crude product is reprecipitated from a mixture of ethyl acetate an~ petroleum ether and the 3-acetoxymethyl-7a-methoxy-7~-[2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetylamino3-3-cephem-4-carboxylic acid tert.-butyl ester thus obtained is filtered off and dried; thin layer chromatogram (silica gel): Rf =
0.48 (system: ethyl acetate/chloroform/acetic acid, 80:19:1);
ultraviolet absorption spectrum (in 95% strength aqueous ethanol): AmaX = 243 m~ ( ~ = 15,800); infrared absorption spectrum (in methylene chloride): characteristic bands at 2.88 ~, 2.93 ~, 5.61 ~l, 5.75 ~, 5.83 ~ and 6.66 ~.
The starting material can be prepared as follows:

~ 3 _,9~ _ ,~ i , .

i.(~SL~

8.15 g of 2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetic acid are dissolved in 500 ml of methylene chloride containing 3~06 ml of N-methylmorpholine, the solution, from which moisture is kept excluded, is cooled to -20C and 4.13 ml of chloroformic acid isobutyl es-ter are added drop-wise whilst keeping the temperature at between -15C and -20C. After 30 minutes, a solution of 9.85 g of 3-acetoxy-me-thyl-7~-amino-3-cephem-4-carboxylic acid tert.-butyl ester in 50 ml of methylene chloride is added, after which the solution is stirred for a further 15 hours whilst slowly raising the temperature to 20C to 25C. me mixture is poured out into ice-cold water, neutralised with dipotassium hydrogen phosphate and extracted repeatedly with methylene chloride. The organic extracts are washed with an aqueous saturated sodium chloride solution and dried over magnesium sulphate, after which the solvent is removed under reduced pressure. The residue is recrystallised froQ diethyl ether and gives 3-acetoxymethyl-7~-[2-(5-tert.-butoxycarbonylamino-methyl-2-thienyl)-ace-tylamino]-3-cephem-4-carboxylic acid tert.-butyl ester, melting poin-t 76-78C, thin layer chromato-gram (silica gel): Rf = 0.44 (system: chloroformlethyl acetate/acetic acid, 80:19:1).
Example 9:
A solution of 3.80 g of 3-acetoxymethyl-7a-methoxy-7~-[2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetyl-amino]-3-cephem-4-carboxylic acid tert.-butyl ester in 15 ml-of trifluoroacetic acid is left to stand for 30 minutes and evaporated, with addition of 50 ml of toluene, under a ~Y

~ .
waterpump vacuum. The residue is triturated with 30 ml of diethyl ether and filtered. The fil-ter residue, which con-tains the trifluoroacetic acid salt of 3-acetoxymethyl-7~-[2-(5-aminome~hyl-2-thienyl)-acetylamin~-7~-methoxy-3-cephem-4-carboxylic acid, is converted, as described in Example 4, into the inner salt of 3-acetoxymethyl-7~-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-cephem-4-carboxylic acid.
Example 10:
1.55 g of 3-acetoxymethyl-7~-[2-(3-tert.-butoxy-carbonylaminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylic acid tert.-butyl ester are reacted with 0.065 g of lithium in 20 ml of methanol and 0.31 ml of tert.-butyl hypochlorite i~ accordance with the method described in Example 8, and 3-acetoxymethyl-7a-methoxy-7~-[2-(3-tert.-butoxycar-bonylaminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylic acid tert.-butyl ester is thus obtained; thin layer chromato-gram (silica gel): Rf = 0.52 (system: toluene/ethyl acetate, 3:2); ultraviolet absorption spectrum (in 95% strength aqueous ethanol): ~max = 241 m~ (~` = 14,200); infrared absorption spectrum (in methylene chloride): characteristic bands at 2.88 ~, 2.93 ~, 5.59 ~, 5.74 ~I, 5.83 ~ and 6.64 ~.
me starting material can be prepared as follows:
A mixture of 75.0 g of 3-bromomethyl-thiophene and 78.5 g of potassium phthalimide in 1,000 ml of dimethylformamide is stirred for 90 minutes at 100C. The reaction mixture is cooled, poured out onto a mixture of ice and water and extracted four times with a total of 1,500 ml of ethyl acetate.
The organic extracts are washed with water, dried over _ ,g~ _ . . ~
~ !

l()S55~;~4 magnesium sulphate and concen-trated under reduced pressure, whereupon 3-phthaloylaminomethyl-thiophene crystallises out;
thin layer chromatogram (silica gel): Rf = 0.78 (system:
toluene/ethyl acetate, 4:1); ultraviolet absorption spectrum (in 95% strength aqueous ethanol): ~max = 220 m~
44,000); infrared absorption spectrum (in methylene chloride):
characteristic bands at 5.59 ~, 5.86 ~ and 6.20 ~.
90.8 g of 3-phthaloylaminomethylthiophene are taken up in 250 ml of acetic arlhydride, 250 ml of trifluoroacetic acid are added dropwise whilst excluding moisture and the mix-ture is stirred for 3 hours at 50C. The reaction solution i.s then concentrated under reduced pressure, 500 ml of water are added and the mixture is neutralised with a 2 N aqueous sodium hydroxide solution whilst cooling with ice, and is extracted with ethyl acetate. The organic phase is washed with water, dried over magnesium sulphate and concentrated.
Mixed crystals consisting of 2-acetyl-3-phthaloylamino-methyl-thiophene and 2-acetyl-4-phthaloylaminomethyl-thiophene are obtained; th n layer chromatogram (silica gel): Rf =
0.53 and 0.45 (system: toluene/ethyl acetate, 4:1).
38.0 g of the mixture of 2-acetyl-3-phthaloylamino-methyl-thiophene and 2-acetyl-4-phthaloylaminomethyl-thiophene are dissolved in 1,500 ml of methanol, 65 g of thallium-(III) nitrate trihydrate are added under a nitrogen atmosphere and the mixture is stirred for 6 hours at 50C. The reaction mixture is cooled to 5-10C, neutralised with a 2 N aqueous sodium hydroxide solution, concentrated under reduced pressure and extracted with 1,000 ml of chloroform. The organic ~ .
_~_ 105~ 4 extract is washed with a saturated aqueous sodium chloride solution, dried over magnesium sulphate and concentrated.
A mixture of 2-(3-phthaloylaminomethyl-2-thienyl)-acetic acid methyl ester and 2-(4-phtha]oylaminomethyl-2-thienyl)-acetic acid methyl ester is obtained; thin layer chromatogram (silica gel): Rf = 0.59 and 0.54 (system: toluene/ethyl acetate, 4:1).
21 g of the mixture of 2-(3-phthaloylaminomethyl-2-thienyl)-acetic acid methyl ester and 2-(4-phthaloylamino~
methyl-2-thienyl)-acetic acid methyl ester are dissolved in 400 ml of dioxane, 50 ml of a 2 N aqueous sodium hydroxide solution are added and the mixture is stirred for 2 hours at 20C to 25C. It is concentrated to a volume of about 200 ml, diluted with 200 ml of water, acidified with 20%
strength aqueous phosphoric acid and extracted three times wi-th a total of 400 ml of ethyl acetate. me extracts are dried over magnesium sulphate and evaporated under reduced pressure.
- 3.0 g of hydrazine hydrate are added to the evapora-tion residue which has been taken up in 250 ml, and the mix-ture is warmed to 60C for 3~ hours and is then evaporated.
m e residue is taken up in 200 ml of dioxane and reacted with 75 ml of a 2 N aqueous sodium hydroxide solution and 5.0 ml of tert.-butoxycarbonyl azide. The mixture is stirred for 20 hours at 20C -to 25C, concentrated to a volume of about 100 ml, diluted with 100 ml of water and extracted with 200 ml of ethyl acetate. The aqueous solution is adjusted to pH 2 wi-th about 50 ml of 20% strength aqueous phosphoric acid.
~1 ._~ _ . f~- -.
. ..

lVSS~Z4 The precipitate i5 filtered off, the filter residue is washed with ethyl acetate and the filtrate is extracted with three times 200 ml of ethyl acetate. The organic extracts are washed with a saturated aqueous sodium chloride solution, dried over magnesium sulphate solution andconcentrated under reduced pressure. The residue is dried under reduced pressure at 30C and a mixture of 2-(3-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetic acid and 2-(4-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetic acid is obtained, which can be separated by chromatography on 200 g of silica gel, using a mixture of chloroform/ethyl aceta-te/ace-tic acid (80:19:1) as the migra-ting agent, thin layer chromatogram:Rf = 0.44and 0.34respectively (system: chloroform/ethyl acetate/acetic acid, 80:19:1).
However, in the next stage the mixture of the two compounds is used.
4.0 g of the mixture of 2-(3-ter-t.-butoxycarbonyl-aminomethyl-2-thienyl)-acetic acid and 2-(4-tert.-butoxy-carbonylaminomethyl-2-thienyl)-acetic acid are dissolved in r 125 ml of methylene chloride, the solution is cooled to -20C
and 1.80 ml of N-methylmorpholine and 0.864 ml of formic acid isobutyl es-ter are added whilst excluding moisture. After 30 minutes, a solution of 1.97 g of 3-acetoxymethyl-7~-amino-3-cephem-4-carboxylic acid tert.-butyl ester in 10 ml of methylene chloride is added, after which the reaction is allowed to go to completion over the course of 2 hours at -10C and 14 hours at 20C. The mixture is diluted wi-th water and the organic layer is separated off and washed with a saturated aqueous sodium chloride solution. It is dried ~Y
_ .~ _ 1~5~

over magnesium sulphate and evaporated under reduced pressure.
The crude product is chromatographed on 200 g of silica gel.
A 4:1 mixture of methylene chloride/ethyl acetate is used to elute first 3-acetoxymethyl-7~-[2-(3-tert.-butoxycarbonyl-aminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylic acid tert.-butyl ester, thin layer chromatogram (silica gel): Rf =
0.55 (system: toluene/ethyl acetate, 6:4); ultraviolet absorption spec-trum (in 95% strength aqueous ethanol): ~max =
241 m~ (~ = 14,000), infrared absorp-tion spectrum (in mineral oil): characteristic bands at 2.97 ~, 3.02 ~, 5.62 ~1, 5.72 ~, 5.83 ~, 5.96 ~, 6.05 ~, 6.49 ~ and 6.69 ~j proton resonance spectrum (chloroform-d): characteristic signals for the two protons on the thiophene ring: o = 6.94 and 7.17 (AB/J = 5.5) Thereafter, the same solvent mixture is used to elute 3-acetoxymethyl-7~-[2-(4-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylic acid tert.-butyl ester, thin layer chromatogram (silica gel): Rf = 0.41 (system: toluene/ethyl acetate, 6:4); ultraviolet absorption spectrum (in 95% strength aqueous ethanol~: ~max = 242 m~
('_ = 14,200); infrared absorption spectrum (in mineral oil):
characteristic bands at 2.99 ~, 5.62 ~, 5.73 ~, 5.83 ~ and 6.59 ~; proton resonance spectrum (chloroform-d): character-istic signals for the two protons on the thiophene ring: ~ =
6.86(s) and o = 6.99(s).
Example 11:
A solution of 1.5 g of 3-acetoxymethyl-7a-methoxy-7~-[2-(3-tert.-butoxyc~rbonylaminomethyl-2-thienyl)-acetyl-amino]-3-cephem-4-carboxylic acid ter-t.-butyl ester in 6.0 ml _,~_ ,.~

11~5S~4 :
of trifluoroacetic acid is converted in accordance with the process described in Example 4 in-to the inner salt of 3-acetoxymethyl-7~-[2-(3-aminomethyl-2--thienyl)-acetylamino]-7a-methoxy-3-cephem-4-carboxylic acid, thin layer chromato-gram (silica gel): Rf = 0.27 (system: n-bu~tanol~acetic acid/
water, 45:45:10); ultraviolet absorption spectrum (in water):
~max = 237 m~ (~ = 12,700); infrared absor~tion spectrum (in mineral oil): characteristic bands at 5.64 ~, 5.57 ~, 5.97 ~ and 6.55 ~; proton resonance spectrum (formic acid d2): characteristic AB-system at ~ = 7.17 and 7.38 (J = 5.5) of the two hydrogen substituents on the disubstituted thio-phene ring.
Example 12:
3-Acetoxymethyl-7a-methoxy-7~-[2-(4-tert.-bu-toxy-carbonylamino~ethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylic acid tert.-butyl ester is prepared in accordance with the process described in Example 10; thin layer chromatogram (silica gel): Rf = 0.37 (system: toluene/
~ethyl acetate, 3:2); ultraviolet absorptlon spectrum (in 95%
strength aqueous ethanol): ~max = 243 m~ ( ~ = 14,300);
infrared absorption spectrum (in methylene chloride): charac-teristic bands at 2.89 ~; 2.93 ~; 5.60 ~i; 5.75 ~; 5.84 and 6.65 ~.
Example 13:
A solution o~ 1.5 g of 3-acetoxymethyl-7~-methoxy-7~-[2-(4-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetyl-amino]-3-cephem-4-carboxylic acid tert.-butyl ester in 6.0 ml of trifluoroacetic acid is conver-ted, analogously to the

9'J
_ ~ _ 10559i~4 process described in Example 4, into the inner salt of 3-acetoxymethyl-7~-[2-(4-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-cephem-4-carboxylic acid, thin layer chromato-gram (silica gel): Rf = 0.27 (system: n-butanol/acetic acid/
water, 45:45:10); ultraviolet absorption spectrum (in water), ~max =238 m~(~ = 13,100); infrared absorption spectrum (in mineral oil): characteristic b&nds at 5.64 ~; 5.75 ~;
5.98 ~ and 6.55 ~.
Exam~le 14:
A lithium methoxide solution prepared by dissolving 0.14 g of lithium wire in 20 ml of methanol is added to a solution, first introduced into the vessel under a nitrogen atmosphere, at -70C, of 4.48 g of 7~-[2-(5-tert.-butoxy-carbonylaminomethyl-2-thienyl)-acetylamino]-3-(1-methyl-5-tetrazolyl-thiomethyl)-3-cephem-4-carboxylic acid diphenyl-methyl ester in 100 ml of tetrahydrofurane and 100 ml of methanol. Immediately thereafter, 0.860 ml of tert.-butyl hypochlorite is added, the mixture is stirred for 35 minutes at -70C to -75C, 3.0 ml of acetic acid are added and the mixture is concentrated under reduced pressure. 50 ml of ice-cold water are added to the residue; the aqueous mixture is extracted with twice 100 ml of ethyl acetate. The orga-nic ex-tracts are washed with a saturated aqueous sodium chloride solution, dried over magnesium sulphate, filtered through 10 g of silica gel and concentrated. me residue is triturated with petroleum ether and 7~-[2-(5-tert.-bu-toxy-carbonylaminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-(1-methyl-5-tetrazolylthiomethyl)-3-cephem-4-carboxylic acid -- 1.04 --~5~4 diphenylmethyl ester is filtered off, thin layer chromato-gram (silica gel): Rf = 0.26 (system: toluene/ethyl acetate, 3:2); ultraviolet absorption spectrum (in 95% strength aqueous ethanol): inflections a-t 239 m~ ( = 13,100) and 276 m~ (& = 5,800); lnfrared absorption spectrum (in mineral oil): charac-teristic bands at 2.90 ~, 5.60 ~, 5.78 ~ and 6.59 ~-The starting material can be obtained, for example,as follows:
2.09 g of p-toluenesulphonic acid monohydrate are added to a suspension of 5.0 g of 7~-amino-3-(1-methyl-5-tetrazolylthiomethyl)-3-cephem-4-carboxylic acid (USA Patent No. 3,516,997) in 200 ml of methanol, whilst stirring; this slowly produces a clear solution, which is concentrated under reduced pressure. Diethyl ether is added and the p-toluene-sulphonic acid salt of 7~-amino-3-(1-methyl-5-tetrazolyl-thiomethyl)-3-cephem-4-carboxylic acid is filtered off. The salt is dissolved in 100 ml of dioxane and 2.5 g of diphenyl-diazomethane are added in portions to the solution. The mixture is stirred for 18 hours at room temperature and con-centrated under reduced pressure; 100 ml of an 0.1 N aqueous sodium bicarbonate solution are added to the residue and the mixture is extracted with twice 100 ml of ethyl acetate.
The organic extracts are washed successively with water and with a saturated aqueous sodium chloride solution, dried over magnesium sulphate and concentrated. The crystallisation from diethyl ether gives 7~-amino-3-(1-methyl-5-tetrazolyl-thiomethyl)-3-cephem-4-carboxylic acid diphenylmethyl ester, _~_ ~()555~;~4 thin layer chromatogram (silica gel): Rf = 0.21 (system:
chloroform/ethyl ace-tate/acetic acid, 80:19:1); ultraviolet absorption spectrum (ethanol): AmaX =269 m~(~ = 6,400);
infrared absorption spectrum (in methylene chloride): charac-teristic bands at 2.93 ~, 5.60 ~ and 6.15 ~.
- 5.43 g of 2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetic acid are dissolved in 200 ml of methylene-chloride containing 2.20 ml of 4-methyl-morpholine, the solu-tion, from which moisture is kept excluded, is cooled to -20C and 2.86 ml of chloroformic acid isobutyl ester are added dropwise. After 30 minutes, a solution of 9.35 g of 7~-amino-3-(1-methyl-5-tetrazolylthiomethyl)-3-cephem-4-carboxylic acid diphenylmethyl ester in 100 ml of methylene chloride is added, after which stirring is continued for one hour at -20C and for 16 hours whilst slowly warming to room temperature. The mixture is poured out onto ice-cold water, the organic solution is separated off and the aqueous phase is again extracted with 100 ml of methylene chloride. The organic extracts are washed with saturated aqueous sodium chloride solution, dried over magnesium sulphate and concen-trated. Crystallisation of the residue from a mixture of methyl acetate and diethyl ether gives 7~-C2-(5-tert.-butoxy-carbonylaminomethyl--2-thienyl~-acetylamino]-3-(1-methyl-5-tetrazolylthiomethyl)-3-cephem-4-carboxylic acid diphenyl-methyl ester, thin layer chromatogram (silica gel): Rf =
0.38 (system: chloroform/ethyl acetate/acetic acid, 80:19:1);
ultraviolet absorption spectrum (in 95% strength aqueous ethanol): shoulders at 242 m~ ( = 16,400) and 274 m~

Ci ~ ... _ . .

lV5~ 4 9,200); infrared absorp-tion spectrum (in mineral oil):
characteristic bands at 3.03 ~, 5.58 ~1, 5.79 ~, 5.96 ~, 6.12 and 6.51 ~.
The starting material can also be prepared in the following manner:
1.5 ml of triethylamine, 1.6 ml of N,N-dimethy]aniline and 2.3 ml of trimethylchlorosilane are added to a suspension of 2.0 g of 7~-amino-3~ methyl-5-tetrazolylthiomethyl)-3-cephem-4-carboxylic acid in 60 ml of methylene chloride. The mixture is warmed to 40C for 20 minutes, whilst stirring,and is then cooled to -10C. To this mixture is added a solution of 2-(5-tert.-bu-toxycarbonylaminomethyl-2-thienyl)-acetic acid chloride in me-thylene chloride (which is prepared as follows:
a solution of 1.95 g of 2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetic acid in 20 ml of me-thylene chloride is treated with 0.05 ml of dimethylformamide and 5 ml of oxalyl chloride. The mixture is stirred under a stream of ni-trogen for 30 minutes at the reflux -temperature, the readily volatile constituents are distilled off under reduced pressure and the oily residue is dissolved in 20 ml of methylene chloride).
The reaction mixture is stirred for 2 hours at 0C to 25C
and poured into water, -the mixture is adjusted to a pH value of about 7 with sodium bicarbonate, extracted with methylene chloride and acidified c~utiously with concentrated hydro-chloric acid and the precipitate formed is filtered off. The filter residue is washed with water and dried under reduced pressure at room temperature.
The product, containing 7~-[2-(5-tert.-butoxy-75~
_ ,,~ _ ~ l)5~ 4 carbonylaminomethyl-2-thienyl)-acetylamino]-3-(1-methyl-5-tetrazolylthiomethyl) 3-cephem-4-carboxylic acid, is dissolved in 30 ml of dioxane, 1.5 g of diphenyldiazomethane are added to the solution and the mixture is stirred for 12 hours at room temperature and then concentrated. After crystal-lisation from a mixture of methyl acetate and diethyl ether, the residue gives 7~-[2-(5-tert.-butoxycarbonylaminome-thyl-2-thienyl)-acetylamino]-3-(1-methyl-5-tetrazolylthiomethyl)-3-cephem-4-carboxylic acid diphenylmethyl ester.
Example 15:
.~ solution of 3.91 g of 7~-[2-(5-tert.-butoxycarbonyl-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-(1-methyl-5-tetrazolylthiomethyl)-3-cephem-4-carboxylic acid diphenyl-methyl ester in 30 ml of trifluoroacetic acid and 5 ml of anisole is left to stand for one hour at 20C with exclusion of moisture and is then concentrated under reduced pressure.
The residue is dried for 16 hours under a hlgh vacuum and is then dissolved in 30 ml of methanol. The solution is treated with an active charcoal preparation and filtered. About 0.95 ml of triethylamine is added dropwise to the filtrate until a pH value of 6.0 is reached; the inner salt of 7~-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-(1-methyl-5-tetrazolylthiomethyl) 3-cephem-4-carboxylic acid, which precipitates, is fil-tered off; thin layer chromatogram (silica gel): Rf = 0.10 (system n-butano'/acetic acid/water, 45:45:103; ultraviolet absorption spectrum (in 0.01 N aqueous sodium bicarbonate): AmaX = 240 m~ (~, = 15,900); infrared absorption spectrum (in mineral oil): characteristic bands -- ,a~
~ !

1()5~ 4 at 5.67 ~l, 5.96 ~ and 6.48 ~.
The following compounds are prepared analogously on selecting the corresponding starting materials: 7~-~2-(5-aminomethyl~2--thienyl)-acetylamino]-3,7a-dimethoxy-3- -cephem-4-carboxylic acid; 3-aminocarbonyloxyme-thyl-7~-[2-(5-aminomethyl~2-thienyl)-acetylamino]-7a-methoxy-3-cephem-4-carboxylic acid; 7~-[2-(5-aminomethyl-2-thienyl)-acetyl-amino]-3-methylaminocarbonyloxymethyl-7a-methoxy-3-cephem-4-carboxylic acid; 7~-[2-(5-aminomethyl-2-thienyl)-acetyl-amino]-3-N-(2-chloroethyl)-aminocarbonyloxymethyl-7a-methoxy-3-cephem-4-carboxylic acid; and 7~-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-methylthiomethyl-3-cephem-4-carboxylic acid, which are usually obtained in the form of their inner salts.
Example 16:
Dry ampoules or phials, containing 0.5 g of the inner salt of 3-acetoxymethyl-7~-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-cephem-4-carboxylic acid, are prO-duced as follows:
Composition (for 1 ampoule or phial) 3-Acetoxymethyl-7~-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-cephem-4-carboxylic acid, inner salt 0.5 g Mannitol 0.05 g A sterile aqueous solution of the inner salt of 3-acetoxymethyl-7~-~2-(5-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy~3-cephem-4-carboxylic acid and of the mannitol is sealed in 5 ml ampoules or 5 ml phials under aseptic '6;.~ - ~ -. .

;~
conditions, and tested.
Example 17.
Dry powders or phials, containing 0.5 g of the inner salt of 7~-[2-(5-aminomethyl-2-thienyl)-ace-tylamino]-7~-methoxy-3-(l-methyl-5-tetrazolyl-thiomethyl)-~-cephem-4-carboxylic acid are prepared as follows:
Composition (for 1 ampoule or phial) 7~-[2-(5-Aminomethyl-2-thienyl)-acetylamino]-7~-methoxy-3-(1-methyl-5-tetrazolyl-thiomethyl)-3-cephem-4-carboxylic acid, inner salt 0.5 g Mannitol O.05 g A sterile aqueous solution of the inner salt of 7~-[2-(5-aminomethyl-2~thienyl)-acetylamino]-7~-methoxy-3-(l-methyl-5-tetrazolyl-thiomethyl)-3-cephem-4-carboxylic acid and of the mannitol is subjected -to freeze drying under aseptic conditions in 5 ml ampoules or 5 ml phials and the ampoules or phials a~e sealed and tested.
Example 18-A solu~Dn of 1.96g of 7~-~2-(5-tert.-butyloxycarbonyl-aminomethyl-2-thienyl~-acetylamino]-3-(5-methyl-1,3,4-thiadia-zol-2-ylthiomethyl)-3-cephem-4-carboxylic acid tert.-butyl-ester in 150m~ oftetrahydrofuran is treated at -70 with a solution of 0.073 g of lithium in 20 ml of methanol and immediately thereafter with 0.391 ml of tert.-butylhypo-chlorite. After a reaction tim~ of 15 minutes at -70 3 ml of acetic acid are added and the reaction mixture is evaporated under about 11 mm Hg. pressure. The residue is taken up in ~ _ ~05~

150 ml of ethyl ac~tate; the organic solution is washed with a dilute aqueous sodium thiosulfate solution and with a saturated aqueous sodium chloride solution, dried over magne-s~m sulfate and concentrated to a volume of about 10 ml under about 11. mm Hg. pressure. The concentrated solution is slowly diluted with 100 ml of petroleum ether; the residue is filtered off and dried under high vacuum at room temperature to yield the 7~-[2-(5-tert.-butyloxycarbonylaminomethyl-2-thienyl)-acetylamino~-7a-methoxy-3-(5-methyl-1,3,4-thiadiazol-2-ylthio-methyl)-3-cephem-4-carboxylic acid tert.-butyl ester; thin layer chromatography (silicagel): Rf = 0,12 (system: toluene/
ethyl acetate 3:2).
The starting material is prepared as follows:
A solution of 4.08 g of 2-(5-tert.-butyloxycarbonylaminomethyl-2-thienyl)-acetic acid in 200nl of methylene chloride containing 1.65 ml of 4-methyl-morpholine is cooled to -20 under exclusion of moisture and treated dropwise with 1.95 ml of chloro-formic acid isobutyl ester. After 30 minutes a solution of 5.16 g of 7~-amino-3-~5-methyl-1,3,4-thiadiazol-2-ylthio-methyl)-3-cephem-4-carboxylic acid and 7.20 ml N,0-bis-(tri-methylsilyl)-acetic acid amide in 150nlof methylene chloride is added, whereupon the reaction mixture is stirred during one hour at -20 and during four hours while the temperature is allowed to slowly rise to room temperature and is then evapo-rated under reduced pressure. The residue is dissolved in water with the addition of sodium hydrogen carbonate until the pH
reaches 8. The solution is washed with ethyl acetate, the ` _ ~ _ lVS~
separated aqueous layer is acidifed to pH 2 witll aqueous phosplloric acid of 20 % strength and is extracted with ethyl acetateO The organic extract is washed with a saturated aqueous sodium chloride solution, dried over magnesium sulfate and evaporated under reduced pressure. One thus obtains the 7~-[2-(5-tert.-butyloxycarbonylaminomethyl-2-thienyl)-acetyl-amino]-3-(5-methyl-1,3,4-thiadiazol-2-ylthiomethyl)-3-cephem-4-carboxylic acid; thin layer chromatogram (silicagel): ~f = 0,40 (system: n-butanoL~acetic acid/water 45:45:10~, ultraviolet absorption spectrum (ethanol):-~rlax 244 m~ ( = 16900);
infrared absorption spectrum (in mineral oil): characteristic bonds at 2,93 ~, 5,60 ~, 5,90 ~, and 6.55 ~
A mixture of 6,1 g of dicyclohexylcarbodiimide, 2,1 g of tert.-butanol and 0,06 g a cop~E~I)-chloride is stirred for five days at room temperature. The resulting suspension of the O-tert.-butyl-N,N'-dicyclohexyl-isourea is diluted with 30 ml of methylene chloride and added to a solution of 2,0 g of 7~-[2-(5-tert.-butyloxycarbonylaminomethyl-2-thienyl)-acetyl-aminol-3-(5-methyl-1,3,4-thiadiazol-2-ylthiomethyl)-3-cephem-4-carboxylic acid in 50 ml methylene chloride, kept at room tem-perature. After five hours the reaction mixture is filtered and the filtrate is evaporated under reduced pressure. The residue is chromatographed using 40 g of silicagel; with ethyl acetate the 7~-[2-(5-tert.-butyloxycarbonylaminomethyl-2-thienyl~-acetylamino]-3-(5-methyl-1,3,4-thiadia~ol-2-ylthio-methyl)-3-cephem-4-carboxylic acid tert.-butyl ester is eluted;
thin layer chromatogram (silicagel): ~f = 0,70 (n-butanol/acetic .. ,, ~
,.~.;,. -_;~ _ 1~5 acid/wa~er 45:45~
Example 19 A solution of 1,52 g of 7~-[2-(5-tert.-butyloxycar-bonyl~minomethyl-2-thienyl)-acetylamino]-7~-methoxy-(5-methyl-1,3,4-thiadiazol-2-ylthiomethyl)-3-cephem-4-carboxylic acid tert.-butyl ester in 20 ml of trifluoroacetic acid is allowed to stand for 15 minutes at room temperature and under ex-clusion of moisture and,after adding toluene, is then evaporated under a pressure of about 11 mm Hg. The residue is tri-turated with diethyl ether; the resulting trifluoroacetic acid salt of the 7~-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-(5-methyl-1,3,4-thiadiazol-2-ylthiomethyl)-~-cephem-4-carboxy-lic acid is dried under high vacuum and at room temperature, then dissolved in 20 ml of water. The aqueous solution is washed with ethyl acetate, adjusted to pH 6 by adding triethyl-amine and concentrated to a volume of about 5 ml under a pressure of about 11 mm Hg. The concentrate is diluted drop-wisewith 30 ml of acetone the reaction mixture is allowed to stand for 2 hours at 4 and the precipitate is filtered off.
The latter is washed with diethyl ether and dried under high vacuum at room temperature. One thus obtains the inner salt of the 7~-[2-(5-aminomethyl-2-thienyl)-acetylamino~-7a-methoxy-3-(5-methyl-1,3,4-thiadiazol-2-ylthiomethyl)-3-cephem-4-carboxy-lic acid, thin layer chromatogram (silicagel): Rf = 0,11 (system: n-butanol/acetic acid/water 45:45:10 ); ultraviolet absorption spectrum (O,l-n. aqueous sodium hydrogen carbonate solution): ~ max = 242 m~ ( - 16400); infrared abso-rption ,. /oC) _~ _ 1(~55~i~4 spectrum (in m;neral oil): characteristic bonds at 5,66 ~, 5,97 ~, 6,25 ~ and 6,50 ~.

Claims (28)

THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:

1. Process for the manufacture of 7.beta.-acylamino-7.alpha.-methoxy-3-cephem-4-carboxylic acid compounds of the formula (I) wherein X rerpresents sulphur or oxygen, or represents etheny-lene of the formula -CH=CH-, and wherein R1 represents lower alkoxy or a radical of the formula -CH2-R2, wherein R2 denotes a hydroxyl or mercapto group etherified by lower alkyl, or heterocyclylthio, wherein heterocyclyl represents a monocyclic five-membered heterocyclic radical of armomatic character which is bonded to the thiosulphur atom via a ring carbon atom and which contains 2 or 3 ring nitrogen atoms and optionally addition-ally a ring oxygen atom, ring sulphur atom or ring nitrogen atom, and such a radical can optionally be substituted by lower alkyl, or wherein R2 denotes lower alkanoyloxy, optionally N-lower alkylated or N-halogeno-lower alkylated carbamoyloxy, and R represents hydroxyl or an etherified hydroxyl group which toghether with the carbonyl grouping -C(=o)-forms an esterified carboxyl group which can be split under physiological conditions, and of pharmaceutically acceptable salts thereof, characterised in that in a compound of the formula (II) wherein the amino group can optionally be substituted by a group which permits the acylation, and Rl has the above mentioned meaning and wherein R0 has the meaning of R or represents a carboxyl protective radical which forms a pro-tected carboxyl group with the carbonyl group of the formula -C(=O)-, or in a salt thereof, the amino group is acylated by treatment with an acid of the formula (III) wherein the amino group is protected if appropriate, or with a reactive functional acid derivative thereof or with a salt of such a compound, or the methoxy group is introduced into the 7.alpha.-position of a 3-cephem compound, of the formula (IV) wherein amino of the aminomethyl group is preferably in the protected form and Ro and Rl have the abovementioned meaning, a carboxyl group of the formula -C(=O)-Ro being in a protected form, or of a salt thereof, by treating an acylimino compound of the formula (IVa) wherein amino of the aminomethyl group is in a protected form, and Ro and Rl have the abovementioned meaning, and a carboxyl group of the formula -C(=O)-Ro is in a protected form, with methanol or by treating a compound of the formula IV, wherein amino of the aminomethyl group and the carboxyl group of the formula -C(=O)-R , and additional functional groups which may be present, are in a protected form, with an anion-forming agent followed by a N-halogenating agent and, if required, reacting with a base which splits off hydrogen halide, and treating the resulting compound with methanol or by treating a compound of the formula (IVb) wherein R° represents an organic radical and wherein amino of the aminomethyl group and the carboxyl group of the formula -C(=O)-Ro, and additional functional groups which may be pre-sent, are in a protected form, with halogen, followed by a base, these reactions being carried out in the presence of methanol, or with methanol in the presence of a desulphurisation agent, or in a compound of the formula (V) wherein AmO represents a protected amino group and Rlo denotes a radical which together with the carbonyl grouping of the formula -C(=O)- forms an unprotected or protected carboxyl group, and wherein amino in the aminomethyl group is in a pro-tected form which differs, in respect of its method of conversion into the free amino group, from that of the protected amino group AmO, the group Am is converted into the free amino group, and the 5-amino-5-carboxy-valeryl radical is split off under the reaction conditions, or a compound of the formula (VI) wherein Ro and Rl have the abovementioned meaning and a carboxyl group of the formula -C(=O)-Ro is in a protected form, is reacted with a compound of the formula R -NH2 (VII), wherein Rx denotes an amino protective group, and formaldehyde or a reactive derivative thereof, in the presence of a strong, at most slightly nucleophilic acid, or a 2-cephem compound of the formula (VII) wherein amino of the aminomethyl group and/or a carboxyl group of the formula -C(=O)-Ro is in a protected form, if required or desired, is isomerised to the corresponding 3-cephem compound by treating it with a weakly basic agent and isolating the corresponding 3-cephem compound from an equilibrium mixture obtained, or by oxidising it in the l-position by means of a per-acid, and, if desired, separating an isomer mixture, which can be obtained, of the l-oxides of corresponding 3-cephem compounds, and reducing the l-oxides, thus obtainable, by means of a reducing agent, and in a resulting compound protected amino of the aminomethyl group is converted into free amino and, if required or desired, a carboxyl group of the formula -C(=O)-R0 is converted into a carboxyl group of the formula -C(=O)-R and/or, if desired, a group R1 is converted into an-other group R1 and/or, if desired, a resulting salt is converted into the free compound or into a pharmaceutically acceptable salt, or a resulting free compound is converted into a pharmaceutically acceptable salt.

2. Process according to Claim 1, characterised in that a protected carboxyl group of the formula -C(=O)-R0 in a starting material is an easily splittable esterified carboxyl group, wherein Ro represents an etherified hydroxyl group.

3. Process according to Claim 1, characterised in that in a starting material free functional groups which may be present in addition to a carboxyl group of the formula -C(=O)-R0 are present in an easily splittable protected form.

4. Process according to Claim 1, characterised in that in a starting material amino of the aminomethyl group is protected by easily removable amino protective groups or is present in the form of the azido group.

5. Process according to Claim 1, characterised in that the acylation of a starting material of the formula II is effect-ed by treatment with an acid of the formula III in the presence of a condensation agent.

6. Process according to Claim 1, characterised in that the acylation of a starting material of the formula II is effected by treatment with an anhydride, including a mixed or inner hydride, of an acid of the formula III.

7. Process according to Claim 6, characterised in that the mixed anhydride used is the anhydride of such an acid with a hydrogen halide acid, hydrazoic acid, an acid containing phosphorus or sulphur, hydrocyanic acid, with a lower alkane-carboxylic acid which is optionally substituted by halogen, or with a half-ester of carbonic acid.

8. Process according to Claim 1, characterised in that the acylation of a starting material of the formula II is effected by treatment with an activated ester of an acid of the formula III.

9. Process according to Claim 1, characterised in that the anion-forming agent used is an alkali metal alcoholate, which base can at the same time serve as an agent which splits off hydrogen halide, and the N-halogenating agent used is a sterically hindered organic hypohalite.

10. Process according to Claim 9, characterised in that the alkali metal alcoholate used is lithium methylate and the sterically hindered organic hypohalite used is tert-butyl hypochlorite.

11. Process according to Claim 1, characterised in that R° in a starting material of the formula IV b represents methyl.

12. Process according to Claim 1, characterised in that a starting material of the formula IV b according to Claim 1 is reacted with chlorine, followed by tri-lower alkylamine.

13. Process according to Claim 1, characterised in that the desulphurisation agent used is a silver or mercury oxide or a silver-I or mercury-II salt.

14. Process according to Claim 1, characterised in that the reactive derivative of formaldehyde used is a polymer thereof.

15. Process according to Claim 1, characterised in that the strong, at most slightly nucleophilic,acid used is trifluoroacetic acid.

16. Process according to Claim 1, characterised in that the per-acid used for oxidising a 2-cephem compound of the formula VIII is 3-chloro-per-benzoic acid and the reducing agent used for reducing the l-oxides of the resulting 3-cephem compounds thus obtainable, is phos-phorous trichloride.

17. Process according to Claim 1, 6 or 9, characterised in that in a resulting compound of the formula I, wherein the amino group in the aminomethyl substituent may be protected, and wherein Rl denotes a group of the formula -CH2-R2 and R2 represents a lower alkanoyloxy group, R2 is converted into another group R2 as defined in claim 1.

18. Process according to Claim 1, 6 or 9, characterised in that in a compound of the formula I, wherein Rl denotes a group of the formula -CH2-R2 and R2 represents a lower alkanoyloxy group, or in a salt thereof, such a radical R2 is replaced by a heterocyclylthio group R2 as defined in Claim 1 by treatment with a corresponding heterocyclic mercaptan compound.

19. Process according one of Claims 1, 6 or 9, characterised in that a compound of the formula IV according to Claim 1, or a salt thereof, wherein X represents sulphur or oxygen and the aminomethyl-substituted radical denotes aminomethyl-2-thienyl or 2-furyl, and wherein Ro has the meaning indicated in Claim 1 and R
represents lower alkoxy with up to 4 carbon atoms or the radical of the formula -CH2-R2, wherein R2 denotes acetoxy, carbamoyloxy, N-lower alkylcarbamoyloxy, N-halogeno-lower alkyl-carbamoyloxy, thiadiazolylthio or tetrazolylthio which are optionally substituted by lower alkyl and bonded to the thiosulphur atom by a ring carbon atom, and wherein amino of the aminomethyl group and a carboxyl group of the formula -C(=O)-Ro may be in a protected form, is reacted with an anion-forming agent followed by a base which splits off hy-drogen halide, and the resulting compound is treated with methanol or in that a compound of formula IVa, wherein X, Ro and Rl have the meaning indicated above and wherein the aminomethyl group and the group -C(=O)-Ro may be protected as above, is reacted with methanol or in that a compound of formula II, wherein Ro and Rl have the same meaning as in-dicated above and in which the group -C(=O)-Ro may be protected, is acylated by treatment with an acid of the formula III, wherein X has the meaning indicated above and wherein the amino group may be protected, or with a reactive functional acid derivative thereof or with a salt of such a compound and after each of above reactions the remaining protective groups are split off to give corresponding 3-cephem compounds of the formula I, or salts thereof, wherein X, R
and Rl have the meaning indicated above and wherein R re-presents hydroxyl.

20. Process according to Claim 1, 3 or 9, characterised in that a com-pound of the formula IV according to Claim 1, or a salt thereof, wherein X
represents sulphur or oxygen and the aminomethyl-substituted radical denotes aminomethyl-2-thienyl or -2-furyl, R has the meaning indicated in Claim 1, and wherein Rl represents methoxy or the radical of the formula -CH2-R2, wherein R2 denotes acetoxy, carbamoyloxy, methylcarbamoyloxy, 2-chloroethyl-carbamoyloxy, methylthio, 5-methyl-1,3,4-thiadiazol-2-ylthio or 1-methyl-5-tetrazolythio is reacted with an anion-forming agent followed by a base which splits off hydrogen halide, and the resulting compound is treated with meth-anol and after the reactions the remaining protective groups are split off to give corresponding 3-cephem compounds of the formula I, or salts thereof, wherein X and Rl have the meaning indicated above and wherein R? represents hydroxyl.

21. Process according to Claim 1, 4 or 6, characterised in that 3-acetoxy-methyl-7.beta. amino-7.alpha.-methoxy-3-cephem-4-carboxylic acid, 3-(1-methyl-S-tetrazol-ylthiomethyl)-7.beta. amino-7.alpha.-methoxy-3-cephem-4-carboxylic acid, or 3-(5-methyl-1,3 J 4-thiadiazol-2-ylthiomethyl)-7.beta. amino-7a-methoxy-3-cephem-4-carboxylic acid diphenylmethylesters are reacted with 2-(5-aminomethyl-2-thienyl)-acetic acid, 2-(5-aminomethyl-2-furyl)-acetic acid, 2-(3-aminomethyl-2-thienyl)-acetic acid or 2-(4-aminomethyl-2-thienyl)-acetic acid, wherein the amino of the aminomethyl group is protected by a tert.-butyloxycarbonyl protecting group, in the presence of a condensation agent, or with a salt of such an acid in the presence of a chloroformic acid lower alkyl ester, or with a mixed or inner anhydride or with an activated ester of such an acid and remaining protecting groups being split off to give 3-acetoxymethyl-7.beta. [2-(5-aminomethyl-2-thienyl)-acetylamino]-7.alpha.-methoxy-3-cephem-4-carboxylic acid, 7 .beta. -2-(5-amino-methyl-2-thienyl)-acetylamin] -7.alpha.-methoxy-3-(1-methyl-5-tetrazolylthiomethyl)-3-cephem-4-carboxylic acid, 3-acetoxymethyl-7 .beta. [2-(5-aminomethyl-2-furyl)-acetylamino]-7.alpha.-methoxy-3-cephem-4-carboxylic acid, 7.beta. C2-(5-amino-methyl-2-furyl)-acetylamino]-7.alpha.-methoxy-3-(5-methyl-1,3,4-thiadiazol-2-ylthiomethyl)-3-cephem-4-carboxylic acid, 3-acetoxymethyl-7 .beta. -[2-(5-aminomethyl-2-furyl)-acetylamino]-7.alpha.-methoxy-3-cephem-4-carboxylic acid, 7 .beta. [2-(5-amino-methyl-2-furyl)-acetylamino] -7.alpha.-methoxy-3-(5-methyl-1,3,4-thiadiazol-2-ylthiomethyl)-3-cephem-4-carboxylic acid, 3-acetoxymethyl-7.beta. [2-(3-aminomethyl-2-thienyl)-acetylamino]-7.alpha.-methoxy-3-cephem-4-carboxylic acid or 3-acetomymethyl-7 .beta. [2-[4-aminomethyl-2-thienyl)-acetylamin] -7.alpha.-methoxy-3-cephem-4-carboxylic acid or salts thereof.

22 Process according to one of claims 1, 3 or 9, characterized in that 3-acetoxymethyl- .beta.- 2-(5-aminomethyl-2-thienyl)-acetylimino]-3-cephem-4-carboxylic acid, 7.beta. -[2-(5-aminomethyl-2-thienyl)-acetylimino]-3-(l-methyl-5-tetra-zolylthiomethyl)-3-cephem-4-carboxylic acid, 3-acetoxymethyl-7.beta.-[2-(5-aminomethyl-2-furyl)-acetylimino]-7.alpha.-methoxy-3-cephem-4-carboxylic acid, 3-acetoxymethyl-.beta.-[2-(3-aminomethyl-2-thienyl)-acetylimino]-3-cephem-4-carboxylic acid or 3-acetoxy-methyl-.beta.-[2-(4-aminomethyl-2-thienyl)-acetylimino]-3-cephem-4-carboxylic acid or salts which are protected at the amino and/or at the carboxylic group, are reacted with methanol, and remaining protecting groups being split off to give 3-acetoxymethyl- .beta.-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7-methoxy-3-cephem-4-carboxylic acid, .beta.-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7a-methoxy-3-(1-methyl-5-tetrazolyl-thiomethyl)-3-cephem-4-carboxylic acid, 3-acetoxymethyl-7b-[2-(5-aminomethyl-2-furyl)-acetylamino]-7a-methoxy-3-cephem-4-carboxylic acid, 3-acetoxymethyl-7.beta.-[2-(3-aminomethyl-2-thienyl)-acetylamino]-7.beta.-methoxy-3-cephem-4-carboxylic acid or 3-acetoxymethyl-7.beta. -[2-(4-aminomethyl-2-thienyl)-acetylamino]-7.alpha.-methoxy-3-cephem-4-carboxylic acid or salts thereof, the starting compounds acetoxymethyl-being optionally prepared by reacting the corresponding 7~-(substituted) acetylamino cephem compounds of formula IV with lithiummethylate, and then with tert.-butyl hypochlorite.

23. Process according to claim 1, wherein the sodium salt of 3-acetoxy-methyl-7~-~2-~5-aminomethyl-2-thienyl)-acetylamino]-7~-methoxy-3-cephem-4-carboxylic acid and the sodium salt of 5-mercapto-1-methyl-tetrazole are re-acted so as to prepare the inner salt of 7~-[2-~5-aminomethyl-2-thienyl)-acetylamino]-7~-methoxy-3-~1-methyl-5-tetrazolylthiomethyl)-3-cephem-4-car-boxylic acid or a pharmaceutically acceptable salt thereof.

24. Process according to one of Claims 1, 9 and 10, wherein diphenyl-methyl 7~-[2-(S-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetylamino]-3-Cl-methyl-5-tetrazolylthiomethyl)-3-cephem-4-carboxylate is reacted with tert.-butyl-hypochlorite in the presence of lithium methoxide and methanol and in the resulting diphenylmethyl 7~-[2-~5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetylamino]-7~-methoxy-3-(1-methyl-5-tetrazolylthiomethyl)-3-cephem-4-carboxylate the diphenylmethyl group is split off, so as to prepare the inner salt of 7~-[2-(5-aminomethyl-2-thienyl)-acetylamino]-7~-methoxy-3-(l-methyl-5-tetrazolylthiomethyl)-3-cephem-4-carboxylic acid or a pharma-ceutically acceptable salt thereof.

25. Process according to one of Claims 1, 9 and 10, wherein diphenyl-methyl 3-acetoxymethyl-7~-[2-C5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylate is reacted with tert.-butylhypochlorite in the presence of lithium methoxide and methanol and in the resulting diphenyl methyl 3-acetoxymethyl-7~-methoxy-7~-[2-(5-tert.-butoxycarbonylamino-methyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylate the protecting groups are split off, so as to prepare the inner salt of 3-acetoxymethyl-7~-methoxy-7~-~2-(5-aminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylic acid or a pharmaceutically acceptable salt thereof.

26. Process according to one of Claims 1, 6 and 7, wherein 2-~5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetic acid is treated with isobutyl chloroformate in the presence of a base or with phosgene in dimethyl formamide and the resulting acid derivative is reacted with diphenyl methyl 3-(1-methyl-5-tetrazolyl-thiomethyl-7.beta.-amino-7.alpha.methoxy-3-cephem-4-carboxylate and in the resulting diphenyl methyl 3-(1-methyl-5-tetrazolylthiomethyl)-7.alpha.-methoxy-7.beta.-[2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylate the protecting groups are split off, so as to prepare the inner salt of 3-(1-methyl-5-tetrazolylthiomethyl)-7.alpha.-methoxy-7.beta.-[2-(5-aminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylic acid or a pharma-ceutically acceptable salt thereof.

27. Process according to one of Claims 8 and 9, wherein 2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetic acid is treated with isobutyl chloroformate in the presence of a base or with phosgene in dimethyl form-amide and the resulting acid chloride is reacted with diphenyl methyl 3-(1-methyl-5-tetrazolyl-thiomethyl-7.beta.-amino-7.alpha.-methoxy-3-cephem-4-carboxylate and in the resulting diphenyl methyl 3-(1-methyl-5-tetrazolylthiomethyl)-7.alpha.-methoxy-7.beta.-[2-(5-tert.-butoxycarbonylaminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylate the protecting groups are split off, so as to prepare the inner salt of 3-(1-methyl-5-tetrazolylthiomethyl)-7.alpha.-methoxy-7.beta.-[2-(5-aminomethyl-2-thienyl)-acetylamino]-3-cephem-4-carboxylic acid or a pharmaceutically acceptable salt thereof.

28. Compounds of the formula I

(I) wherein X represents sulphur or oxygen, or represents ethenylene of the formula -CH=CH-, and wherein R1 represents lower alkoxy or a radical of the formula -CH2-R2, wherein R2 denotes a hydroxyl or mercapto group etherified by lower alkyl, or heterocyclylthio, wherein heterocyclyl represents a monocyclic five-membered heterocyclic radical of aromatic character which is bonded to the thiosulphur atom via a ring carbon atom and which contains 2 or 3 ring nitrogen atoms and optionally additionally a ring oxygen atom, ring sulphur atom or ring nitrogen atom, and such a radical can optionally be substituted by lower alkyl, or wherein R2 denotes lower alkanoyloxy, optionally N-lower alkylated or N-halogeno-lower alkylated carbamoyloxy, and R represents hydroxyl or an etherified hydroxyl group which together with the carbonyl grouping -C(=O)-forms an esterified carboxyl group which can be split under physiological conditions, or pharmaceutically acceptable salts thereof, whenever prepared by the process of claim 1 or an obvious chemical equivalent thereof.