GB2045233A - Unsaturated 3-heterocyclyl- thiomethyl-7???-methoxy-7???- acylamido-3-cephem-4- carboxylic acid derivatives - Google Patents

Unsaturated 3-heterocyclyl- thiomethyl-7???-methoxy-7???- acylamido-3-cephem-4- carboxylic acid derivatives Download PDF

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GB2045233A
GB2045233A GB7910345A GB7910345A GB2045233A GB 2045233 A GB2045233 A GB 2045233A GB 7910345 A GB7910345 A GB 7910345A GB 7910345 A GB7910345 A GB 7910345A GB 2045233 A GB2045233 A GB 2045233A
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cephem
vinylene
methoxy
thiomethyl
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Farmitalia Carlo Erba SRL
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D501/00—Heterocyclic compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
    • C07D501/14—Compounds having a nitrogen atom directly attached in position 7
    • C07D501/16—Compounds having a nitrogen atom directly attached in position 7 with a double bond between positions 2 and 3
    • C07D501/20—7-Acylaminocephalosporanic or substituted 7-acylaminocephalosporanic acids in which the acyl radicals are derived from carboxylic acids
    • C07D501/57—7-Acylaminocephalosporanic or substituted 7-acylaminocephalosporanic acids in which the acyl radicals are derived from carboxylic acids with a further substituent in position 7, e.g. cephamycines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents

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  • Cephalosporin Compounds (AREA)
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Abstract

A compound having the general formula (I> <IMAGE> wherein Z is cyano, carbamoyl or -COOR wherein R is hydrogen or C1-C6alkyl; X is a free or esterified carboxy group; A is <IMAGE> wherein Y is a') hydrogen; b') C1-C6 alkyl; c') C2-C5 alkenyl; d') -(CH2)n-CN wherein n is an integer of 1 to 4; e') -(CH2)n-COOR, wherein n and R are as defined above; f') -(CH2)n-CONH2, wherein n is as defined above; or <IMAGE> wherein R1 is a'') hydrogen; b'') C1-C5 alkyl; c''> <IMAGE> wherein each of the groups R, which may be the same or different, is as defined above; d'')-NHCONH2; e'') -NHCH2COOR; f'') -COOR; g'') -CH=CH-COOR; h'') -(CH2)m-COOR; or i'') -(CH2)m-CN; wherein R is as defined above and m is zero or an integer of 1 to 3, and the pharmaceutically or veterinarily acceptable salts thereof have high anti-bacterial activity and are useful in treating infections caused by micro-organisms. They can be incorporated into pharmaceutical compositions.

Description

SPECIFICATION Unsaturated 3 - heterocyclyl - thiomethyl - 7a methoxy - 7ss - acylamido - 3 - cephem -4 - carboxylic acid derivatives The present invention relates to unsaturated - 3 heterocyclyl - thio - methyl - 7 - methoxy - 7,8 - acylamido - 3 - cephem - 4 - carboxylic acid derivatives, to a process for their preparation and to pharmaceutical and veterinary compositions containing them.
The compounds of the invention have the following general formula (I)
wherein Z is cyano, carbamoyl, -COOR wherein R is hydrogen or C,-C6 alkyl; X is a free or esterified carboxy group; Airs: a)
wherein Y is a') hydrogen; b') 01-06 alkyl; c') C2-Cs alkenyl; d') -(CH2)n-CN wherein n is an integer of 1 to 4; e') -(CH2)n - COOR, wherein n and R are as defined above; f') -(CH2)n - CONY2, wherein n is as defined above; or b)
wherein R1 is a") hydrogen;
wherein each of the groups R, which may be the same or different, is as defined above; d") -NHCONH2; e")-NHCH2COOR; f") -COOR; g") -CH=CH-COOR; h") -(CH2)m-COOR; i") -(CH2)m - CN; wherein R is as defined above and m is zero or an integer of 1 to 3.
The present invention also includes the pharmaceutically orveterinarily acceptable salts of the compounds of formula (I) as well as all the possible isomers, for example cis- and trans-isomers and stereoisomers, and their mixtures, the metabolites provided with antibacterial activity and the metabolic precursors of the compounds of formula (I).
The alkyl and the alkenyl groups may be branched or straight chain.
Because of the presence of the -CH=CH- group on the 7,3-chain the compounds of formula (i) may be both cis (or Z) isomers and trans (or E) isomers as well as a mixture thereof; when another -CH=CH- group is presentthen the compound of formula (I) may be eithercis, cis-isomers or trans, transisomers orcis, trans-isomers or trans, cis-isomers or mixtures thereof.
When one or more asymmetric carbon atoms are present, the compounds of formula (I) may be either optically active or racemic compounds. A C,-C6 alkyl group preferably is methyl, ethyl, n-propyl, or n-butyl.
A C2-C6 alkenyl group preferably is vinyl or allyl. The group -CH=CH- on the 7,8chain preferably is cis CH=CH-.
When X is an esterified carboxy group it is preferably a group of formula -COOR2 wherein R2 is one of the radicals
wherein R is as defined above, R3 is a basic group, in particular an alkyl, for example C,-C6 alkyl, or an aralkyl, for example benzyl, group substituted by at least an amino group, which in turn may be unsubstituted or substituted (for instance R3 may be C1-C6 alkyl - NH - OH3, C1-C6 aralkyl -NH-CH3, C1-C6 alkyl
R4 is an alkyl group, in particular a C1-C6 alkyl group, e.g. methyl, propyl or isopropyl; an aryl group, in particular phenyl; a cycloalkyl group, in particular cyclopentyl, cyclohexyl and cycloheptyl; a heteromonocyclic group, for example pyridyl; a bicyclic group, for example indanyl; an aralkyl group, for example benzyl. The pharmaceutically and veterinarily acceptable salts of the compounds of formula (I) are those either with inorganic acids, such as hydrochloric and sulphuric acid, or with organic acids, such as citric, tartaric, malic, maleic, mandelic, fumaric and methanesulphonic acid, or with inorganic bases, such as sodium, potassium, calcium, or aluminium hydroxides and alkaline or alkaline-earth carbonates or bicarbonates, or with organic bases, such as organic amines, e.g., lysine, trietylamine, procaine, dibenzylamine, N - benzyl -,3 -phenethylamine, N, N'-dibenzylethylenediamine, N - ethyl - piperidine, diethanolamine, N - methylglucamine, tris - hydroxymethyl - aminomethane and the like.
Also the internal salts (i.e. zwitterions) are included in the scope of the invention.
Preferred salts are those of the compounds of the invention wherein X is a free carboxy group with inorganic bases, e.g., those mentioned above.
Preferred compounds of the invention are the cisisomers of the compounds of formula (I) wherein Z is cyano, carbamoyl or-COOR wherein R is hydrogen or C1-C6 alkyl; X is a free or salified carboxy group and A is:
wherein Y is -CH3; -(CH2)n-CN; -(CH2)n-COOH or-(cH2)n-coNH2 wherein n in an integer of 1 to 4; or wherein R, is hydrogen or amino; and the pharmaceutically or veterinarily acceptable salts thereof.
Specific examples of the compounds of the invention are the following: 1) 7ss -[2 - cyano - vinylene (Z) - thioacetamidoj - 7&alpha;- methoxy-3-[(1H-1,2, 3, 4-tetrazol-5-yl)- thiomethyl] - 3 - cephem - 4 - carboxylic acid; 2) 7,3 - [2 - cyano - vinylene (Z) - thioacetamidoj - 7a - methoxy-3-[(1- methyl - 1,2,3,4 - tetrazol - 5 - yl) thiomethyl] - 3 - cephem - 4 - carboxylic acid; 3) 7ss - [2 - cyano - vinylene (Z) - thioacetamido] - 7&alpha; methoxy - 3 - [(1 - ethyl - 1,2,3, 4 - tetrazol - 5 - yl) - thiomethyl] - 3- cephem - 4 - carboxylic acid; 4) 7ss -[2 - cyano - vinylene (Z) - thioacetamido] - 7&alpha; - methoxy-3-[(1 - carboxy - methyl -1, 2, 3, 4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 5) 7ss - [2- cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - [(1 - ss - carboxyethyl - 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 6) 7ss - [2- cyano - vinylene (Z) - thioacetamido] - 7ae - methoxy - 3 -[(1 - y- carboxypropyl - 1,2,3,4 tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7) 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] - 7a methoxy - 3 - [(1 - (2) - propenyl - 1, 2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 8) 7ss -[2 - cyano - vinylene (Z) - thioacetamido] - 7 - methoxy - 3- [(1 - carboxamidomethyl - 1,2,3,4 tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 9) 7ss -[2- cyano - vinylene (Z) - thioacetamido] - 7&alpha; - methoxy - 3 -[(1 - ss - carboxamidoethyl -1, 2, 3, - tetrazol -5 - yl) - thiomethyl] - 3- cephem - 4 - car- boxylic acid; 10)7,3 - [2- cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - [(1 - - carboxamidopropyl - 1,2,3,4 tetrazol - 5 - yl) - thiomethyl] - 3 - cephemcarboxylic acid; 11) 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] 7a methoxy - 3 -[(1 - ss - cyano ethyl - 1,2,3,4 - tetrazol 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid;; 12)7,3 - [2- cyano - vinylene (Z) - thioacetamido] - 7a - methoxy-3 -[(1-7- cyanopropyl - 1,2,3,4- tetrazol - 5 - yl) - thiomethyl] - 3- cephem - 4 - car- boxylic acid; 13)7,3 -[2- carboxamido - vinylene (Z) thioacetamidod - 7a - methoxy - 3- [(1 - methyl - 1, 2, 3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 carboxylic acid; 14)7,3 -[2 - carboxamido - vinylene (Z) thioacetamido] - 7a - methoxy - 3 - [(1 - ethyl - 1,2,3, 4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 carboxylic acid;; 15)7,3 - [2- carboxy - vinylene (Z) - thioacetamido] 7a-methoxy-3-[(1 - methyl -1,2,3,4-tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 16) 7,3 - [2 - carboxy - vinylene (Z) - thioacetamido] 7&alpha;- methoxy-3 -[(1-.ethyl-1,2,3,4-tetrazol-5-yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 17)7,3 - [2 - carboxy - vinylene (Z) - thioacetamido] 7&alpha;- methoxy - 3 -[(1 -(2)-propenyl-1,2,3,4- tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - car- boxylic acid;; 18)7,3 - [2- cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - [(tetrazolo [1,5-b] pyridazin - 6 - yl) thiomethyl] - 3 - cephem - 4 - carboxylic acid; 19) 7ss -[2 - carboxamido - vinylene (Z) thioacetamido]- 7a - methoxy - 3- [(tetrazolo [1, 5-b] pyridazin - 6 - yl) -thiomethyl] - 3 - cephem - 4 - carboxylic acid; 20) 7,3 - [2 - carboxy - vinylene (Z) - thioacetamido] 7a - metoxy - 3 - [(tetrazolo [1 , 5-b] pyridazin - 6 - yl) thiomethyl] - 3 - cephem - 4 - carboxilic acid; 21) 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] - 7a methoxy - 3 - [(tetrazolo 1,5-b] pyridazin - 8 amino - 6 - yl) - thiomethyl - 3 - cephem - 4 - carboxylic acid; 22) 7,3 - [2 - carboxamido - vinylene (Z) thioacetamido] - 7a - methoxy - 3 - [(tetrazolo [1, 5-b] pyridazin - 8 - amino - 6 - yl) thiomethyl] - 3 - cephem - 4 - carboxylic acid; 23) 7ss - [2 - carboxy - vinylene (Z) - thioacetamido] 7a - methoxy - 3 [(tetrazolo [1, 5-b] pyridazin - 8 - amino - 6 - yl) - thiomethyfi - 3 - cephem - 4 - carbox- ylic acid, and the pharmaceutically or veterinarily acceptable salts thereof.
The structural formulae of the above numbered compounds, indicated according to their progressive number are reported in the following table.
T A B L E
compound Z X A 14-N .
1 NC- -COOH q N N / 2 NC- COO N ln/ CH3 N-N 3 NC- -COON \C,Bg 4 NC- -COOH N ' N' CH2CoOH N-N S NC- -COOH N; N' CH 2CH2 COOH 6 NC- -COON "\(CH,!f-CooH 2 3
compound Z X A NN 7 NC- COOil- N / \CH,,-C'I=CH2 N-N 8 1 NC- -CoOlI N; N \ \ CH2CONH2 N 9 - NC- -CO^:I ì (CH A CO:IH .
10 NC- -COO H N 4 '(Cl )-CO11 23 2 . NN M/ \ (CH95 2CN - ----- -- 12 ;ic- 2 A' (Ci1);CN 13 1 H21-=- JtNA N
compound Z X A N-N 14 N2N-CO- -COH A N \ C 2H5 N N 15 ilOOC- COOH N/ Ecí3 3 16 ilOOC- -COON 'C N 25 Nl4 17 HOOC- -COOH JN/N XC112CH=CH2 18 NC- -COON ~N N 19 | H2N-CO- -COOH 20 flCOC- -COON .
compound Z X A NH2 ,4"t. N 21 NC- -COOH . toH2 22 H, > N-CO- -COOH 23 HOOC- -COON NH2 Nhi N~~ The compounds of the invention are prepared by a process comprising: A) reacting a compound of formula (II)
wherein Z and A are as defined above and X' is a free or salified or esterified carboxy group, with a compound of formula M-OCH3 wherein M is an alkaline metal in the presence of a positive halogenating agent; or B) reacting a compound of formula (Ill)
wherein Z and X' are as defined above, with a compound of formula (IV) HS-A (lav) wherein A is defined above, or a reactive derivative thereof; or C) reacting a compound of formula (V)
Wherein X' and A are as defined above and E is amino or a group -N=C=O wherein 0 is an oxygen or sulphur atom, or a reactive derivative thereof, with an acid of formula (VI) Z-CH=CH-S-CH2-COOH (Vl) wherein Z is as defined above, or a reactive derivative thereof; or D) reacting a compound of formula (all)
wherein X' and A are as defined above and Y is halogen, with a compound of formula (VIII) Z-CH=CH-SH (veil) wherein Z is as defined above, or a reactive derivative thereof; or E) reacting a compound of formula (IX)
wherein X' and A are as defined above, or a reactive derivative thereof, with a compound of formula (X) Z-CH=CH-Y' (X) wherein Z is as defined above and Y' is halogen or the residue of a reactive ester of an alcohol; or F) reacting a compound of formula (XI)
wherein Z, X' and A are as defined above, with methyl alcohol in the presence of Hg++ or Ag ions; or G) removing the acylating aminoadipoyl group in a compound of formula (XII)
wherein Z, A and the group X', independently, are as defined above and T is an amino protecting group and, if desired, converting a compound of formula (I) into a pharmaceutically or veterinarily acceptable salt thereof and/or, if desired, obtaining a free compound from a salt and/or, if desired, converting a compound of formula (I), or a salt thereof, into another compound of formula (I), ora saltthereof, and/or, if desired, resolving a mixture of isomers into the single isomers.
Analogously to what reported above for compounds (I) also the starting materials used in the above processes A to G, when the only -CH=CH- group in the 7,3chain is present may be eithercis-isomers or trans-isomers or mixtures thereof; when two groups -CH=CH- are present they may be eithercis, cisisomers ortrans, trans-isomers orcis, trans-isomers or trans, cis-isomers or mixtures thereof; when one or more asymmetric carbon atoms are present, they may be optically active or racemic compounds.
When in the compounds having the formulae (II), (III), (V), (all), (IX), (Xl), and (XII) reported above X' is a free carboxy group, it is preferably protected in a conventional manner before the reaction takes place.
Examples of protecting groups are those usually employed in the synthesis of peptides and cephalosporins, for example benzidryl, trichloroethyl, ter -butyl, p-nitro-benzyl, p-methoxybenzyl and trimethylsilyl.
The other free carboxy groups possibly present in the above compounds may be protected analogously.
The protecting groups are then removed atthe end of the reaction in a known manner, for example by mild acid hydrolysis or by catalytic hydrogenation.
The compounds of formula (I) containing the protecting groups are also included in the object of the invention.
A reactive derivative of a compound of formula (Vl) may be, for example, an acyl halide, an anhydride, a mixed anhydride, an azide a reactive ester or a salt such as, for example a salt with an alkaline or alkaline-earth metal, ammonia or an organic base. A reactive derivative of the compounds having the formulae (IV), (VIII) and (IX) preferably is a salt, for example a salt with an alkaline or alkaline-earth metal.
A reactive derivative of the compound of formula (V) may be, for example, an amine salt, a silyl ester or a metal salt when X is carboxy.
In the compound of formula (VII) Y is preferably chlorine or bromine. When in the compound of formula (X) Y' is halogen, the halogen preferably is chlorine or bromine; when Y' is the residue of an active ester of an alcohol, it is preferably -O-mesyl or -O-tosyl.
The reaction of the compound of formula (II) with the compound of formula M-OCH3 is carried out using preferably about 1,6 molar equivalents of the compound M-OCH3 for each molar equivalent of the compound (II) in the presence of 1-5 molar equivalents of a positive halogenating agent, operating in an aprotic organic solvent such as, e.g., diethyl ether, isopropyl ether, tetrahydrofurane or dioxane at a temperature ranging from about 9000 to about 5000.
The positive halogenating agent may be, for example, ter -butyl -hypochlorite or N-chloro-acetamide, preferablyter-butyl - hypochlorite.
In the compound M-OCH3,M preferably is lithium. If desired the compound M-OCH3 may be prepared in situ from methanol, which in this case may be used as solvent, and an alkaline metal, for example lithium, or a metallorganic compound such as, for instance, a lithium alkyl, e.g. butyl lithium, or a lithium aryl, e.g. phenyl lithium.
The reaction between the compound of formula (III) and the compound of formula (IV), or a salt thereof, is preferably performed in water or in an organic solvent mixable with water or in a mixture of water and an organic solvent; a suitable organic solvent may be, for example, acetone, ethanol, dioxane or tetrahydrofurane. The reaction temperature may vary from about +5"C to about +95"C and the pH is preferably maintained between about 5 and about 7,5. In this purpose, if desired, a buffer may be used such as, for instance, sodium phosphate or sodium acetate.
The reaction between the compound of formula (V) and the compound of formula (Vl) or a reactive derivative thereof may be carried out either at room temperature or under cooling, e.g. from about-70 C to about +40 C, in an appropriate organic solvent such as, for example dichloroethane, acetone, dioxane, tetrahydrofurane, acetonitrile, chloroform, methylene chloride, dimethylformamide and, if necessary, in the presence of an organic base, e.g. N, N - diethylaniline, pyridine or a trialkylamine, or inorganic base, e.g. sodium or potassium bicarbonate, or also in the presence of another acid acceptor such as, e.g., an alkylene oxide, for example propylene oxide, or 4A molecular sieves.
When the compound of formula (VI) is reacted as free acid or as salt with a compound of formula (V) wherein E is amino, the reaction is preferably performed in the presence of a condensing agent, e.g.
dicyclohexylcarbodiimide.
The reaction between the compound of formula (VII) and the compound of formula (VIII) may be carried out at temperatures ranging from about -30 C to about +90"C, preferably at room temperature, in the presence of an inorganic or organic base such as, e.g., sodium or potassium hydroxide, sodium carbonate ortriethylamine, in a suitable solvent which may be, for instance, water or a solvent mixable with water or an anhydrous solvent such as, for example, acetone, chloroform, methanol, ethanol or methylene chloride.
The reaction between the compound of formula (IX), or a reactive derivative thereof, e.g. an alkaline salt, and a compound of formula (X) may be performed either in an aqueous medium in the presence of an inorganic base, or in an organic solvent, preferably an anhydrous organic solvent, e.g. methylene chloride, chloroform, dioxane, in the presence of an organic base; the reaction temperature may vary from about-20 C to about +50"C.
The reaction reported above at the point F) may be performed operating, e.g., in methanol or in a mixture of methanol and an anhydrous inert solvent, such as, e.g., diethyl ether, dimethylformamide, tetrahydrofurane, benzene, acetonitrile, dioxane, at a temperature varying from about-50 C (preferably from -15"C s :1000) to the reflux temperature, and, if desired, in the presence of an acid acceptor such as, e.g., pyridine orAg2O.
Preferably the reaction is carried out under nitrogen atmosphere and it is completed within about one hour, generally within 10-15 minutes. The Hg++ ions used to catalyze the reaction may be obtained from a mercuric salt, e.g. HgCI2 or, preferably, Hg(OCOCH3)2; the Ag+ ions may be obtained from a silver salt, e.g. AgNO3, or from Ag2O.
The amino protecting group Tin the compound of formula (XII) may be one of the amino protecting groups indicated above, for exampletert- butoxycarbonyl or trichloroethoxy - ca rbonyl.
The removal of the acylating aminoadipoyl chain in the compound of formula (XII) may be performed by treatment with a reagent able to remove the amino protecting group T.
The removal of the said protecting group and the simultaneous cleavage of the aminoadipoyl chain may be effected by the conventional methods described in the chemistry of the peptides, preferably by treatment with trifluoroacetic acid and anisole when T is tert-butoxycarbonyl according to the procedures described, e.g., in J. Am. Chem. Soc.
94, 1410 (1972) and in Tetrah. Letters, 3979 (1975).
The optional conversion of a compound of formula (I) into a pharmaceutically or veterinarily acceptable salt thereof as well as the optional conversion of a salt into a free compound may be effected in a conventional way.
Also the optional separation of a mixture of isomers into the single isomers as well as the optional conversion of a compound of formula (I) or a salt thereof into another compound of formula (I) or a salt thereof may be performed in a known manner.
Thus, for example, a compound of formula (I) wherein X is a free or salified carboxy group may be converted into a compound of formula (I) wherein X is an esterified carboxy group by reaction with the suitable halide, in an organic solvent, e.g. acetone, tetrahydrofurane, chloroform, methylene chloride, dimethylformamide, dimethylsulphoxide, or in a mixture of water and an organic solvent, e.g. dioxane or acetone; the reaction temperatures may vary from about-20"C to about +80"C.
A compound of formula (I) wherein X is an esterified carboxy group may be converted into the corresponding compound wherein X is a free carboxy group by treatment, e.g. with an inorganic acid e.g.
hydrochloric or trifluoroacetic acid, as described in the organic chemistry.
Analogous conversions may be performed, if desired, on the other free or esterified carboxy groups possibly present in the compounds of formula (I) wherein, in particular, a free a esterified carboxy group may be converted, if desired, into a carbamoyl group by reaction with ammonia by conventional procedures.
Also the optional resolution of a mixture of isomers into the single isomers may be carried out in a known manner. Thus, for example, racemic compounds may be separated into the single optical antipodes, e.g., by resolution, for instance by fractionate cristallization of the diastereoisomeric salts mixtures and, if desired, liberating the optical antipodes from the separated salts. The separation of the cis- and trans- isomers from their mixture may be performed by fractionate cristallization from a polar solvent, for example an aliphatic alcohol, e.g.
ethanol or a mixture of a polar solvent with water or water, or also by chromatography e.g. column or preparative or thin layer chromatography or HPLC.
The protection of the carboxy groups in the starting materials may be effected by the usual methods employed in the chemistry of peptides and cephalosporins.
Thus, for example, a tert-butyl ester of a free carboxy group may be obtained by reaction with isobutene in dioxane in the presence of concentrated sulphuric acid according to what described in J. Med. Chem. 9, 444 (1966) or, preferably, by reaction with an O-tert-butyl-isourea derivative, preferably 0 - tert - butyl - N, N' - diisopropyl - isourea, in an inert solvent, e.g. methylene chloride, as also known in the literature.A benzhydryl ester of a free carboxy group may be obtained, for example, by treatment with diphenyl - diazo - methane in an inert solvent such as, e.g., acetonitrile, dioxane, tetrahydrofurane, ethyl acetate, around the room temperature, if desired salifying atfirstthe basic groups possibly present by a strong not nucleophilic acid, e.g., perchloric or p-toluen-sulphonic acid. Atrichloroethyl ester of a free carboxy group may be obtained reacting the acid, or a reactive derivative thereof, with 2, 2, 2 trichloroethanol in an inertsolventsuch as, e.g.
methylene chloride possibly in the presence of a base, e.g. pyridine, and/or a condensing agent such as, e.g., dicyclohexylcarbodiimide. A p - metoxybenzyl or p - nitro - benzyl - ester may be obtained, for example, reacting the corresponding halide, for example p - methoxy - benzyl chloride or p - nitro benzyl bromide with a salt of the acid to be esterified in an inert solvent, preferably liquid sulphurous anhydride, as described, e.g., in Tetr. Lett. 2915 (1977).
The compound of formula (II) may be prepared, for example, by the processes B) and C) reported above using as starting materials 7,3 - amino - cephalosporanic acid derivatives instead of 7,8 - amino - 7a methoxy - cephalosporanic acid derivatives.
The compound of formula (III) may be obtained by the same methods reported above at the points A), C), D), E), F), and G) for the preparation of the compounds of formula (I) using as starting materials compounds having the formulae (II), (V), (VII), (IX), (Xl) and (XII) wherein the -CH2-S-A substituent is replaced by the -OCOCH3 group.
The compound of formula (IV) may be prepared by known methods or, respectively, is a known compound.
The compound of formula (V) may be prepared, for example, by the methods described in Tetr. Lett.
2705 (1975) and J. Antibiotics 29, 554 (1976); preferably the compound (V) is prepared by oxidizing a Schiff base of formula (XIII)
wherein X' and A are as defined above and each of R6 and R6, which may be the same or different, is C1-C6 alkyl, preferablytert-butyl, then introducing the -OOH3 group in the 7a - position of the obtained quinoide compound and finally removing the benzyl radical. The oxidation of the compound of formula (XIII) may be performed, for example by treatment with PbO2 or DDQ (dichloro - dicyano - benzoquinone) in a suitable organic solvent such as, e.g., benzene, toluene or chloroform, at temperatures ranging between about-20 C and about +20 C.
The introduction of the 7a-methoxy group in the intermediate quinoide-compound may be carried out, for example, by treatment with methanol at temperatures varying from about 0 C to the reflux temperature.
The final removal of the benzyl radical may be effected, e.g. by treatment with hydrazine or phenyl hydrazine or with a Girard reagent, preferably the Girard T reagent, in an appropriate solvent which may be, for instance, methanol, ethyl acetate, acetone, methylene chloride, dimethylformamide at a temperature ranging from about 0 C to about 50"C.
In the compound of formula (V) so obtained E is amino. Alternatively the methoxylation of the Schiff base of formula (XIII) may be performed by producing the intermediate quinoide-compound by chlorination - dehydrochlorination, instead of by oxidation, for instance by treatment with lithium methoxide and tert - butyl hypochlorite as described, e.g., in J. Antibiotics, 29,969(1966). The compound (V) may be prepared by reacting a compound of formula (XIV)
wherein X' and E are as defined above, with a compound of formula (IV) using approximately the same reaction conditions reported above for the reaction between the compound (III) and the compound (IV).
A compound of formula (V) wherein E is amino may be converted into a compound of formula (V) wherein E is -N=C=, wherein IZI is as defined above, by known methods, for example by reaction with phosgene orthiophosgene in the presence of an hydrochloric acid acceptor, e.g. triethylamine, in an inert solvent such as, for instance, benzene, toluene, tetrahydrofurane or diethyl ether, at a temperature which may vary between the room and the reflux temperature.
The compound of formula (VI) may be prepared by reacting a compound of formula (X) with thioglycolic acid, or a reactive derivative thereof, for instance a salt or an ester, or by reacting the thioglycolic acid, or a reactive derivative thereof, with a compound of formula Z-CCH, wherein Z is as defined above and, when necessary, hydrolysing the obtained ester. In both the cases the reactions are preferably carried out in water or an organic solvent, e.g. tetrahydrofurane, diethyl ether, benzene or a mixture of one of these solvents and water, in the presence of 0, 5-2, 5 equivalents of a base, for example triethylamine, sodium or potassium bicarbonate or sodium or potassium hydroxide.Depending on the reagents and reaction conditions a cis- or a trans- compound of formula (VI) or a mixturecis, trans- is obtained: the second way is the preferred when it is desired to obtain the cis-isomer, expecially if a low temperature is used.
When the reaction product consists in a mixture of cis- and trans- isomers, these may be separated by the methods usually employed in the organic chemistry for the resolution of geometric isomers mixtures, e.g. by fractionate cristallization from a suitable solvent, for instance water or a lower aliphatic alcohol, or by chromatography.
A compound of formula (VI) wherein Z is cyano may be alternatively obtained from the corresponding compound wherein Z is carbamoyl by treatment with a dehydrating agent, e.g. Pals, POCI3 or triphenylphosphine, in an inert organic solvent such as e.g. diethyl ether or dimethylformamide, or by heating at 300C-120"C in an organic solvent preferably chosen from the group of hexamethylenphosphotriamide and dimethylsulphoxide.
The compound of formula (VII) may be prepared by reacting the acid of formula Y-CH2-COOH, wherein Y is as defined above, or a reactive derivative thereof, for example one of the reactive derivatives mentioned above for the compounds of formula (VI) with the compound of formula (V) using approximately the same reaction conditions described above for the reaction between the compound of formula (V) and the compound of formula (VI).The compound of formula (VII) may be obtained, alternatively reacting an acid of formula Y-CH2-COOH, wherein Y is as defined above, or a reactive derivative thereof, preferably an halide, with a compound of formula (XV)
wherein the groups X', independently, T and A are as defined above, so obtaining a compound of formula (XVI)
wherein X', T, Y, and A are as defined above, from which the compound (VII) may be obtained by removal of the acylating aminoadipoyl chain by a procedure analogous to that reported above for the conversion of the compound (XII) into the compound (I).The reaction between the acid Y CH2-COOH, or a reactive derivative thereof, and the compound of formula (XV) is preferably performed in an inert solvent, e.g. methylene chloride, in the presence of an halohydric acid acceptor, e.g. a trimethylsilyl aceta mide, preferably N-trimethylsilyl trifluoroacetamide or N, O - bis - trimethylsilyl trifluoroacetamide, or in the presence of molecular sieves, preferably 4A molecular sieves, according to the methods described, e.g., in J. Am. Chem. Soc.
94, 1410 (1972) or in Chem. Pharm. Bull. (Tokyo),24, 2629(1976).
The compound of formula (VII) may also be obtained, for example, by the process reported in Tetr. Letters 16, 1305(1976), i.e. by a multi-step process comprising: a) reacting a compound of formula (XVII)
wherein X', A and each of the groups Y are as defined above with PCls in an anhydrous inert solvent, e.g. chloroform, methylene chloride or a polychloroethane at temperatures varying from about 6000 to about -30O, in the presence of a base, preferably quinoline, so obtaining a compound of formula (XVIII)
wherein Y, X' and A are as defined above; b) reacting a compound of formula (XVIII) with a compound of formula MOCH3, wherein M is as defined above, preferably lithium, operating at a temperature ranging between approximately -80"C and approximately -70"C under nitrogen atmosphere, in an anhydrous solvent, e.g. diisopropyl ether, tetrahydrofurane or dioxane, mixed with anhydrous methanol, so obtaining a compound of formula (XIX)
wherein Y, X' and A are as defined above: c) hydrolysing a compound of formula (XIX) by known methods, preferably by treatment with trimethylsilylchloride and quinoline at room pressure and room temperature, in an inert solvent according to what described, e.g., in Tetr. Letters, 1307 (1976).
The compounds of formula (VIII) are known compounds or may be prepared by known methods from known compounds.
The compounds of formula (IX) may be prepared reacting a compound of formula (V) with thioglycolic acid, or a reactive derivative thereof, e.g. one of the reactive derivatives indicated above for the acids of formula (VI), using approssimately the same reaction conditions previously reported for the reaction between the compound (V) and the compound (Vl).
The compounds offormula (X) are known compounds. The compounds of formula (Xl) may be prepared reacting a compound of formula (XX)
wherein X' and A are as defined above and R7 and Re are both hydrogen or, taken together, form an arylidene radical, preferably benzylidene, with an acid of formula (VI) or a reactive derivative thereof using reaction conditions analogous to those described above for the reaction between the compound (V) and the compound (VI).
The compounds of formula (XII) may be obtained by reacting a compound of formula (XV) with an acid of formula (VI) or, preferably, a reactive derivative thereof, in the reaction conditions previously reported for the reaction between a compound of formula (XV) and the acid of formula Y-CH2-COOH or a reactive derivative thereof. The compounds of formula (XIII) may be prepared by reaction of a com pound of formula (XXI)
wherein E is amino and X' and A are as defined above with an aldehyde of formula
wherein R5 and R6 are as defined above operating preferably in an inert solvent such as, e.g., benzene, toluene, chloroform at temperatures ranging from room temperature to about 110"C in the presence, if necessary, of a suitable dehydrating agent, e.g.
molecular sieves.
The compounds of formula (XIV) may be prepared: a) starting from a Schiff base of formula (XXII)
wherein X', R5 and R6 are as defined above, by the same process used to obtain the compound (V) from the compound (XIII), i.e. by subsequent oxidation, methoxylation and removal of the benzyl radical, possibly using a chlorination - dehydrochlorination reaction instead of the oxidation; or b) starting from a compound of formula (XXIII)
wherein X', R, and R6 are as defined above, by solvolysis of the methylthio group in the presence of methanol as described above for the conversion of the compound (XI) into the compound (I), that is by reaction with methanol in the presence of Hug+* or Ag+ ions, being the debenzylation of the Schiff base effected either before or after the methoxylation; or c) starting from a compound of formula (XXIV)
wherein X' is as defined above and Ra is an acylating chain stable in the reaction conditions (e.g. to the action of lithium methoxide/tert-butyl hypochlorite) and easily removable at the end of the reaction, for example a p-nitrobenzyloxycarbonyl group, by introducing the methoxy group in the 7a-position in the conditions reported above for the conversion of a compound of formula (II) into a compound of formula (I), then removing the acylating chain Ra as described, e.g., in Tetr.Lett. 1311 (1974); or d) starting from a compound having the formula (XXIV) reported above wherein X' is as defined above and Ra is hydrogen, by diazotization with nitrous acid and addition of bromo-azide to give a 7-bromo-7-azido-cep ha losporanic acid derivative from which a compound of formula (XIV) wherein E is amino may be obtained by exchange of the bromo atom with the methoxy group and by subsequent catalytic hydrogenation of the azido group e.g.
according to the method reported in J. Am. Chem.
Soc. 94,1408(1972).
The compounds of formula (XV) may be obtained from the natural Cephamycin C by a process comprising: a) the preliminary protection of the free amino group by the usual methods described in the chemistry of the peptides and cephalosporins; b) the reaction of the obtained compound with the compound of formula (IV) in the reaction conditions described above for the reaction between the compound (III) and the compound (IV); c) the optional protection of the free carboxy groups present by the methods usually employed in the chemistry of the peptides and cephalosporins.
The compounds of formula (XVII) may be prepared by reacting a compound of formula (XXV)
wherein E, X' and A are as defined above, with an acid of formula (Y)2CH-COOH wherein each of the groups Y, which may be the same or different, is as defined above, using approximately the same reaction conditions described above for the reaction between a compound (V) and a compound (VI).
The compounds of formula (XX) wherein R7 and R8 are both hydrogen may be obtained from the corresponding compounds of formula (XX) wherein R7 and R8, taken together, form and arylidene radical, by mild acid hydrolysis, e.g. with HCI or aqueous H2SO4 or with the Girard T reagent The compounds of formula (XX) wherein R7 and R8, taken together, form a radical arylidene may be in turn prepared, for example, reacting a compound of formula (XXVI)
wherein Ar is aryl and X' and A are as defined above, with a sulphur derivative containing the reactive species CH3-S-6+, such as, e.g., CH3-S-CI, CH3-S-SO2~ CH3 or CH3-S-S-COOCH3, in an anhydrous solvent, e.g. dimethoxyethane, tetrahydrofurane, dioxane, dimethylformamide in the presence of a base such as, for instance, sodium hydride or potassium tertbutylate, at a temperature ranging from about -20 C to a bout -10"C under nitrogen atmosphere as described, e.g., in J. Org. Them.38, 2857 (1973) or in J. Org. Chem. 39, 2794(1974).
The compounds of formula (XXII) may be prepared from 7 - amino - cephalosporanic acid by a process analogous to that reported above for preparing the compounds (XIII) from the compounds (XXI). The compounds having the formulae (XXI), (XXIII), (XXIV), (XXV) and (XXVI) may be easily prepared by known methods starting from known compounds or, respectively, they are compounds already known in literature.
The compounds object of the present invention shown a high antibacterial activity either in animals or in humans against both Gram-positive and Gram-negative bacteria and are therefore useful in the treatment of the infections caused bysaid microorganisms, such as, respiratory tract infections, for example, bronchitis, bronchopneumonia, pleurisy; hepatobiliary and abdominal infections, for example cholecystis, peritonitis; blood and cardiovascular infections, for example pyelonephritis, cystitis; obstetrical and gynecological infections, for instance, cervicitis, endometritis; ear, nose and throat infections, for instance otitis, sinusitis, parotitis.
The following table shows the minimal inhibiting concentrations (MIC) in ,ag/ml of the compounds of the invention identified by the codes K 13163 and K 13204, against Gram-negative bacteria in comparison with Cefazolin and Cefoxitin.
TABLE
Cefazolin Cefoxitin K 13163 K 13204 E. coli Tem '76 12,5 6,2 1,6 1,6 Klebsiella aerogenes 1802E > 100 6,2 2,2 1,6 Enterobacter cloacae 1321 E 3,1 17,8 1,6 0,56 Proteus mirabilis ATCC 9921 6,25 6,25 1,6 0,8 ProteusvulgarisX20 > 100 1,6 0,5 0,2 E. coli GR+ Tem 50 6,2 0,8 0,8 Klebsiella pneumoniae ATCC 10031 0,8 0,56 0,28 0,16 SalmonellatyphiWatson 1,6 3,1 0,5 0,8 Cefazolin = 7,3 - (1 - tetrazolyl - acetamido) - 3 - [2 - (5 - methyl - 1,3,4 - thiadiazolyl) thiomethyl] - 3 - cephem - 4 - carboxylic acid; Cefoxitin = 7,3 -[2 (2 - thienyl) - acetamido] - 7 methoxy - 3 - carbamoyloxymethyl - 3 cephem - 4 - carboxylic acid;; K 13163 = 7,3 -[2 - cyanovinylene (Z) thioacetamido] -7a-methoxy-3-[(1 methyl-1,2, 3, 4-tetrazol-5-yl)- thiomethyl] - 3 - cephem - 4 - carboxylic acid; K 13204 = 7ss - - carboxy - vinylene (Z) thioacetamido- - 7a - methoxy - 3- [(tetrazolo [1, 5-b] pyridazin - 8 - amino - 6 - yl) - thiomethyl] - 3 cephem -4 - carboxylic acid.
As is evident from the table the activity of K 13163 and K 13204 against the strains usually resistant to cephalosporins is higher not only than that of Cefazolin but also than that of Cefoxitin.
Furthermore the compounds of the invention, when tested against the hospital strains usually little sensitive to cephalosporins, e.g. Proteus indole positive bacteria, Serratiae, Citrobacter and Enterobacter, are more active both than Cefoxitin and than Cefazolin.
The compounds of the invention may be administered, either to humans orto animals, in a variety of dosage forms, e.g., orally in the form of tablets, capsules, drops or syrups; rectally in the form of suppositories; parenterally, e.g. intravenously or intramuscolary (as solutions or suspensions), with intravenous administration being preferred in emergency situation; by inhalation in the form of aereosols or solutions for nebulizers; intravaginally in the form, e.g. of bougies; ortopically in the form of lotions, creams and ointments. The pharmaceutical or veterinary compositions containing the compounds of the invention may be prepared in a conventional way by employing the conventional carriers and/or diluents used for the other cephalosporins.
Conventional carriers or diluents are, for example, water, gelatine, lactose, starches, magnesium stearate, talc, vegetable oils, cellulose and the like. Daily doses in the range of about 1 to about 100 mg per Kg of body weight may be used, in various animal species the exact dose depending on the age, weight and conditions of the subjectto be treated and on the frequency and route of administration.
A preferred way of administration of the compounds of the invention is the parenteral one: in this case the compounds may be administered, for example to adult humans, in an amount ranging from about 100 mg to about 200 my pro dose, preferably about 150 mgpro dose, 14 times a day, dissolved in a suitable solvent, such as, for example sterile water or lidocaine hydrochloride solution for intramuscolar injections, and sterile water, physiological saline solution, dextrose solution or the conventional intravenous fluids or electrolytes, for intravenous injections.
Furthermore, the compounds of the invention may be used as antibacterial agents in a prophylatic manner, e.g., in cleaning or as surface disinfecting compositions, for example, at a concentration of about 0,2 to 1% by weight of such compounds admixed with, suspended or dissolved in conventional inert dry or aqueous carriers for application by washing or spraying. They are also useful as nutritional supplements in animal feeds. Assessment of melting points was somewhat difficult in some cases, as the compounds tend to retain the solvent.
In these cases, after the indication of the melting point, the word "dec." (decomposition) was added.
The l.R. spectra were determined in a solid phase on a Perkin-Elmer 125 spectrophotometer, while the U.V. spectra were usually evaluated in a buffer phosphate solution at pH 7.4 on a Bausch-Lomb apparatus. N.M.R. spectra were determined in DMSO (dimethylsulphoxide) with a Bruker HX-90 spectrometer with (CH3)4Si as internal standard.
The following examples illustrate but do not limit the present invention.
EXAMPLE 1 To a solution containing the t-butyl ester of the 7 - (3, 5 - di - t - butyl - 4 hydroxy - benzylidene - amino) - 3 [(1 - methyl - 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] 3 - cephem - 4 - carboxylic acid (11,46g.) in ml 200 of anhydrous tetrahydrofuran (THF) and 85 ml of an hydros ethanol, cooled to -70 C, there is added, with stirring, 38.2 ml of a 0,55 N solution of lithium methoxide in methanol and then a solution contain ing t-butylhypochlorite in 30 ml. of anhydrous s-dichloroethane. The mixture is stirred at -70"C for 50 minutes.After the mixture is broughtto room temperature it is evaporated to dryness under reduced pressure; the residue is taken up with ethyl ether and is filtered; the residue is taken up with 400 ml. of benzene, heated until complete solution takes place, treated with activated charcoal, filtered, and evaporated to dryness, obtaining 9.33 g (78%) of the t - butyl ester of the 7 - ss - (3, 5 - di - t - butyl - 4 - hydroxy - benzylidene - amino) - 3 -[(1 - methyl - 1,2, 3,4 - tetrazol - 5 - yl) - thiomethyl)] - 7 - methoxy - 3 cephem - 4 - carboxylic acid; m.p. 183 - 184"C (with dec.) Elemental analysis (for C30H42N6OsS2) Found: C: 56,75; H: 6.57; N: 13.02; S: 10.10.
Calculated: C: 57.11; H: 6.71; N: 13.32; S: 10.16 I.R. (CH Cm3): v(O-H) 3600 cm-1 v(C=O) p-lactam 1770 cm-t v(C=O) = COO t - Bu - ester 1685 cm-' v(C=N) 1635 cm-' The t - butyl ester of the 7 - (3, 5 - di - t - butyl - 4 - , hydroxy - benzylidene - amino) - 3 -[(1 - methyl - 1,2, 3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid used as the starting material was prepared in the following manner: : A solution containing the t - butyl ester of the 7 amino - 3 -t(1 3, -methyl-1,2,3,4-tetrazol-5-yl)- thio methyl - 3- cephem - 4 - carboxylic acid (3,84) and 4 - hydroxy - 3, 5 - di - tert. butyl - benzaldehyde (2.5 g) in 80 ml of anhydrous benzene was refluxed for one hour, separating the water which is formed during the reaction. The undissolved residue is removed by filtration and the filtrate is evaporated to dryness under reduced pressure.The residue is waken up in cyclohexane, so as to bring in solution the eventual unreacted aldehyde and the resulting solid is collected by filtration, dissolved in benzene and reprecipitated with cyclohexane, obtaining 5.1 g (85%) of the t-butyl ester of the 7 - (3, 5 - di - t - butyl 4 - hydroxy - benzylidene - amino) - 3 -[(1 - methyl - 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem 4- carboxylic acid; m.p. 133-135 (with dec.).
I.R. (CH Cos): v(O-H) 3600 cm-' v(C=O) ss-lactam 1770 cm-' v(C=O)-COO-t-Bu.ester 1685 cm-' v(C=N) 1635 cm-l On the other hand, the t-butyl ester of the 7-amino - 3 - [(1 -methyl-1,2,3,4-tetrazol-5-yl)- thiomethyl] - 3 - cephem - 4 - carboxylic acid was preparated in the following manner: To a suspension containing the 7-amino - 3 -[(1 methyS1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 cephem -4 - carboxylic acid (6,56 g) in 120 ml of methylene chloride, 0 - t - butyl - N, N' - bis - isopropylisourea (14,35 ml) dissolved in 40 ml of methylene chloride is added.The mixture is stirred for4 hours at room temperature and the solid is removed by filtration and washed with methylene chloride. The filtrate is washed with a saturated NaHCO3 aqueous solution, and then with water. The organic layer is separated, dried on sodium sulfate, and evaporated to dryness under reduced pressure.
The oily residue is redissolved with a small amount of chloroform (30 ml) and poured into petroleum ether, obtaining 6,34 g (82%) of the t-butyl ester of the 7 - amino - 3 - [(1 - methyl - 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; m.
m.p.146-150 C l.R. (CH Cm3): v(C=O) ss-lactam 1770 cm-' v(C=O) - COO -t - Bu ester 1685 cm-' EXAMPLE2 To a suspension containing the t - butyl ester of the 7 - ss - (3,5 - di - t - butyl - 4 - hydroxy - ben- zylidene - amino) - 7a - methoxy -3 -[(1 - methyl - 1, 2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 carboxylic acid (6,9 g) in 100 ml of methanol and 100 ml of ethyl acetate there is added with stirring 3,68 g of Girard T reagent dissolved in 70 ml of methanol; the cleavage of the Schiff base is followed by means of thin layer chromatography. The reaction is completed in about 15 minutes.The reaction mixture is concentrated to a small volume, it is taken up in ethyl acetate and washed with water; the water phases are extracted with ethyl acetate and the organic phases are combined, washed with cold water, dried over sodium sulphate and evaporated to dryness under reduced pressure, obtaining 3.88 g (85%) of the t - butyl ester of the 7 - ss - amino - 7a - methoxy-3-[(1 - methyl - 1,2,3,4-tetrazol -5-yl)- thiomethyl] - 3 - cephem - 4 - carboxylic acid: I.R. (CH Cl3):: z'(C=O),3-lactam 1775 cm-' v(C=O) - COO -t- Bu ester 1685 cm-' EXAMPLE3 To a solution of the t - butyl ester of the 7,3 - amino -7a- methoxy-3-[(1 - methyl - 1,2,3,4 - tetrazol -5 -yl(-thiomethyl] -3-cephem4-carboxyl ic acid (4,5 g) in 80 ml of anhydrous sym-dichloroethane, cooled to 0 C, there is added the N, N - diethyl aniline (1,6 ml) and then the 2-cyano-vinylene(Z)mercapto - acetyl chloride (1,61 g) dissolved in 30 ml of sym - dichloroethane. The reaction mixture is then stirred for 30 minutes, diluted with ethyl acetate, washed first with a 3% K H SO4 solution and then with water. It is then washed again with a sodium bicarbonate solution and then with water.
To organic phase is separated, dried over sodium sulfate, filtered and evaporated to dryness under reduced pressure. The semi-solid residue is purified on a silica gel column (300 g), using as an eluent a mixture of ethyl acetate: cyclohexane (2:1). It is then evaporated to dryness, taken up in ethyl ether and filtered, thus obtaining 39 (55,5 %) of the t - butyl ester of the 7 - ss -[2 - cyano - vinylene (Z) thioacetamido] - 7 - a - methoxy - 3 - [(1 - methyl - 1, 2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 carboxylic acid; m.p. 80-85"C 5 (with dec.).
Elemental analysis (for C20H2sNzOsS3) Found: C: 44,69; H: 4,86; N: 17,81; S: 17,52 Calculated: C: 44;50; H: 4,67; N: 18,17; S: 17,82 I.R. (K Br) v(C3N) conjugated 2200 cm-' v(C=O) ss-lactam 1780 cm-' v(C=O) - COO -t - Bu ester 1685 cm-' N.M.R. ppm (C D Cl3) 1,56 (9H, s, -COOt.Bu) 3,40 (3H, s, 7-OCH3) 3,6 (2H, q, 2-OH2) 3,78 (2H, s, -S-CH2-CO)
4,3 (2H, q, 3-OH2) 5,00 (1 H, s,6-H) 5,74(1 H, d, NC-CH=) 7,68 (1 H, d, =CH-S) J CH=CH(Z) = 11 Hz 9,56(1H,s,-CONH).
The 2 - cyano - vinylene (Z) - mercapto - acetyl chloride used as the starting material was prepared in the following manner: To a solution of the 2 - carboxyamido - vinylene (Z) - mercapto - acetic acid (49) (described in Ex. 13) in dimethyl - formamide (20 ml), cooled to 0 C, there is added P 015 (5.2 g), with stirring, maintaining the temperature between 0" and 5 C. After 1 hour of stirring at the same temperature, the reaction mixture is poured in ice and extracted fourtimes with ethyl acetate (4 x 50 ml); the combined extracts are washed with a saturated solution of Na CI, dried over sodium sulfate and then evaporated to dryness at a temperature not higher than 40"C, obtaining a yellow oil which is dissolved in acetone (25ml).
To the resulting solution there is added 6.72 g of dicyclohexylamine, obtaining the precipitation of the dicyclohexylamine salt of the 2 - cyano - vinylene (Z) -mercaptoacetic acid which, after filtration, is washed twice with small quantities of acetone; m.p.
180-183 C.
The salt is then redissolved in H20 - ethyl acetate (5:7; 120ml) at 5"C; the solution is acidified by adding thereto dropwise 40% H3PO4 (10 ml). The resulting solution is extracted three times with ethyl acetate, the organic extracts are washed with water saturated with NaCI, dried over anhydrous sodium sulfate and evaporated to dryness under reduced pressure obtaining the 2 - cyano - vinylene (Z) - mercapto - acetic acid (2,769; 77%); m.p. 90-92 C.
Elemental analysis (for CsHsNO2S): Found: C: 41,70; H: 3,63; N: 9,64; S: 22,25 Calculated: C: 41,94; H: 3,52; N: 9,78; S: 22,39 I.R. (K Br): v(O=N) conjugated 2220 cm-' v(C=O) acid 1720 cm-' N.M.R.(DMSO-d6):5.4(d,NC-CH =);7,4(d, = CHS) JCH = CH (Z) = 10 Hz To a solution containing the 2 - cyano - vinylene (Z) - mercapto - acetic acid (1,43 g) in ethyl ether (70 ml), cooled to 0 C, there is added phosphorus pentach chloride (2,08 g); the mixture is then stirred for 2 hours at 5"C, then evaporated under reduced pressure at a temperature not higher than 40 C, taking up the residue repeatedly with anhydrous benzene and evaporating under reduced pressure, obtaining 1,61 g (100%) of the 2 - cyano - vinylene (Z) - mercapto acetyl chloride, which is sufficiently pure to be used as such in the following reaction.
EXAMPLE4 The t - butyl ester of the 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - {(1 - methyl 1,2,3,4 - tetrazol -5 - yl) - thiomethyl] - 3- caphem 4 carboxylic acid (39) is added to a solution cooled to 0 C containing trifluoroacetic acid (30 ml) and anisole (3 ml), and the mixture is stirred at room temperature for one hour. Both the trifluoroacetic acid and the anisole are removed by evaporation under reduced pressure and the residue is taken up with benzene and evaporated again under reduced pressure, repeating said treatment twice.The residue is dissolved in acetone, and ethyl ether was added, obtaining an oil; the supernatant solution is decanted and the remaining oil is again dissolved in acetone and again reprecipitated with ethyl ether obtaining a solid, which is further purified by chromatography an a silica gel column (809) using as an eluent chloroform; methanol (1:1).
The solution is evaporated under reduced pressure thus obtaining 1,62 g (60%) of 7,3 - [2 - cyano vinylene (Z) - thioacetamido] - 7a - methoxy - 3- [(1 methyl - 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3- cephem -4 - carboxylic acid; m.p. 118 - 120 C (dec.) Elemental analysis (for O16H17N7O5S3): Found: C: 39,97; H:3,71; N: 19,91; S: 19,41 Calculated:C: 39,74; H: 3,54; N: 20,27; S: 19,89 I.R. (K Br): y(O=N) conjugated 2200 cm-' v(C=O)p - lactam 1780 cm-' v(C=O) acid 1630 cm-' v(C-N) + 8 (N-H) sec - amide 1520 cm-t U.V. (buffer phosphate pH 7.4): X,,, = 275 m,a; E11au = 387 T.L.C.Rf=0,67 (CHCI3: OH3OH : HCOOH = 8:2:1) N.M.R. ppm (DMSO-d6): 3,4 (3H, s, 7-OCH3) 3,6 (2H, q, 2-CH2) 3,78 (2H, s, -S-CH2-CO)
4.36 (2H, q, 3-OH2) 5,00 (1 H, s,6-H) 5,74(1 H, d, NC-CH=) 7,68(1 H, d, = CH-S) JCH=CH(Z) = 11 Hz 9,56 (1H, s, -CONH) EXAMPLE 5 To a solution containing the t - butyl ester of the 7 (3, 5 - di - t - butyl - 4 - hydroxy - benzylidene - amino) -cephalosporanicacid(21,76g) in 720 ml of anhydrous tetrahydrofuran and 80 ml of methanol, cooled at-78 C under a nitrogen stream, there are added 80 ml of a solution of 0,55 N lithium methoxide in methanol and then the t - butyl hypochlorite (5,72 ml) dissolved in 80 ml of anhydrous tetrahydrofuran.
The mixture is stirredforforty minutes at-70 C and then concentrated under reduced pressure. The residue is taken up with ethyl ether, triturated, and the undissolved part is then removed by filtration.
The filtrate thus obtained is evaporated to dryness thus obtaining 21 g (93%) of the t - butyl ester of the 7,3 - (3,5 - di - t - butyl - 4 hydroxy - benzylidene amino) - 7a - methoxy - cephalosporanic acid.
Elemental analysis (for C30H42N207S): Found: C: 62.81; H: 7,50; N: 4,51; S: 5,31 Calculated: C: 62,69; H: 7,36; N: 4,87; S: 5;57 I.R. (CH Cl3): v(O-H) 3600 cm-' v(C=O)ss-lactam 1775 cm-' v(C=O) -COOt. Bu-ester 1685 cm-' v(C=N) 1635 cm-l The t - butyl ester of the 7 - (3,5 - di - t - butyl - 4 hydroxy - benzylidene - amino) - cephalosporanic acid used as starting material was preparated in the following manner: A solution containing the t - butyl ester of the 7 amino - cephalosporanic acid (13,12 g) and 4 - hydroxy - 3,5 - di - t - butyl - benzaldehyde (10 g) in 250 ml of anhydrous benzene is refluxed for 1 hour, separating the water which is formed during the reaction.
The solid is removed by filtration, the filtrate is evaporated to dryness and the residue is taken up with ethyl ether. The solid which forms is filtered and the filtrate is evaporated to dryness under reduced pressure obtaining 21,7 g of the t - butyl ester of the 7 - (3,5 - di - t - butyl - 4 - hydroxy - benzylidene amino) - cephalosporanic acid.
I.R. (CH Cl3) = v(O-H) 3600 cm-' v(C=O) ss-lactam 1770 cm-' v(C=O) - COO -t- Bu ester 1685 cm-' v(C=N) 1635 cm-' On the other hand, the t - butyl ester of the 7 amino - cephalosporanic acid was obtained in the following manner: To a suspension of 7 - amino - cephalosporanic acid (16,32g) in 360 ml of methylene chloride there is added the 0 -t - butyl - N, N' - bis isopropyl isourea (36 g) dissolved in 120 ml of methylene chloride. The mixture is then stirred at room temperature for4 hours. The N, N' - bis - isopropylurea separated is filtered off and washed with methylene chloride.The filtrate is washed with a sodium bicarbonate solution and then with water. The organic phase is dried over anhydrous sodium sulfate, filtered and evaporated to dryness. The residue is triturated with cyclohexane and the remaining solid is collected by filtration, obtaining thus 15,8 g (80%) of the t-butyl ester of the 7 - amino - cephalosporanic acid.
I.R. (CH Cm3): i'(C=O),3-Iactam 1770 cm-' v(C=O) - COO - t - Bu ester 1685-' EXAMPLE 6 To a solution containing the t - butyl ester of the 7,3 - (3,5 di - t - butyl - 4 hydroxy - benzylidene - amino) 7a - methoxy - cephalosporanic acid (5,75 g) in 40 ml of methanol there is added with stirring 3,34 g of reactive Girard T and the mixture is stirred for 10 minutes at room temperature; it is taken up with 500ml of ethyl ether and washed with 300 ml of water. The water phases are extracted again with ethyl ether and the organic phase is separated.The ether extracts are combined, washed with water and dried on anhydrons calcium sulfate, filtered and evaporated for dryness obtaining a solid consisting of the t - butyl ester of the 7,3 - amino - 7a- methoxy cephalosporanic acid.
I.R. (CH Cm3): v(C=O) p-lactam 1770 cm-' v(C=O) - COO - t - Bu ester 1685 cm-' EXAMPLE 7 To a solution containing the t-butyl ester of the 7,3 amino - 7a - methoxy - cephalosporanic acid (4.5 g) in anhydrous sym-dichloroethane (80 ml), N, N diethylaniline (1.61 ml) and a solution containing 2 cyano - vinylene (Z) - mercapto - acetyl chloride (1.61 g) (prepared as described in example 3) in anhydrous sym-dichloroethane (30 ml) were added. The mixture was cooled to 0 C, stirred for 30 minutes, diluted with ethyl acetate, washed first with a 3% solution of KHSO4 and then with water. Then it was washed with a sodium bicarbonate solution and again with water.The organic layer was separated, dried over anhydrous sodium sulfate and evaporated to dryness, so obtaining an oil which was purified on a column of silica gel (200 g) using as eluent a mixture of ethyl acetate : cyclohexane (2:1).
The resulting product was then evaporated to dry ness under reduced pressure so obtaining t-butyl ester of the 7,3 - [(2 - cyano - vinylene (Z) thioacetamido] - 7a - methoxy - cephalosporanic acid (2.4 g; 50%).
Elemental analysis (for C20H25N307S2): Found: C: 49,81; H: 5,23; N; 8,39; S: 12,9 Calculated: C: 49,67; H: 5,21; N: 8,69; S: 13,26 I.R. (K Br): v (C-N) Conjugated 2220 cm-' (C=O) ss-lactam 1780 cm-' (C= O) - COO - t - Bu ester 1685 cm-l N.M.R. ppm (ODCI3) 1,56 (9H, s, -COOt.Bu) 2,08 (3H, s, -O-CO-CH3) 3,25(1 H, d, 2-OH2) 3,55 (3H, s, 7-OCH3) 3,57 (1 H, d, 2-OH2) 3,63 (2H, s,-S-CH2-CO) 4,79(1 H, d, -CH2-O-CO-) 5,05 (lH, d, -CH2-O-CO-) 5,07 (1 H, s,6-H) 5,43 (1H, d, NC-CH=) 7,44(1 H, d, =CH-S) J CH=CH'Z) = 11Hz 7,92(1H,s,-CONH) EXAMPLE 8 To a solution of the t - butyl ester of the 7,3 - [(2 cyano - vinylene (Z) - thioacetamido] -7a- methoxy cephalosporanic acid (3,0 g) in anhydrous sym dichloroethane (40ml) there is added anisole (20 ml).
The solution is then evaporated at room temperature so as to eliminate the dichloroethane. To the solution thus obtained, cooled to 0 C, there is added trifluoroacetic acid (40ml) and the solution is then stirred for 1 hour at room temperature; then it is evaporated under reduced pressure to remove the anisole and the trifluoroacetic acid. It is taken up with ethyl acetate (20 ml), treated with activated charcoal, filtered and dicyclohexylamine (0,65 ml) is added. The solution is cooled and the dicyclohexylamine salt precipitated is filtered off, washed with ethyl acetate and then with ethyl ether. The solid is suspended in water, layered with ethyl acetate and acidified with 20% H2SO4. After stirring for 15 minutes, the organic phase is separated, washed with water, dried over Na2SO4, filtered and evaporated to dryness.It is then taken up with ethyl ether obtaining 1,32 g (50%) of 7,3 - [(2 - cyano - vinylene (Z) - thioacetamido] - 7a methoxy - cephalosporanic acid: m.p. 105-110"C (with dec.).
Elemental analysis (for C,6H"N307S2): Found: C: 44,57; H: 4,26; N: 9,53; S: 14,48 Calculated: C: 44,95; H: 4,01; N: 9,83; S: 15,00 I.R. (K Br): V(C-=N) conjugated 2220 cm~' v(C=O) p-lactam 1780 cm-1 v(C=O) conjugated acid 1680 cm-' v(C-N) + 8 (N-H) sec. - amide 1570 cm-' U.V. (buffer phosphate pH 7,4): max=272 my E1%1cm= 383 T.L.C. Rf=0,43 (OHCl3 : OH3OH : HCOOH = 8:1:1) N.M.R. ppm (DMSO - d6) 2,02 (3H, s, -O-CO-CH3) 3,27 (1 H, d,2-CH2) 3,4 (3H, s, 7-OCH3) 3,62 (1 H, d, 2-CH2) 3,76 (2H, s, -S-CH2-CO) 4,67 (1 H, d,1CH2-O-CO-) 4,98(1 H, d, -CH2-O-CO-) 5,16(1 H, s, 6-H) 5,43 (1 H, d, NC-CH=) 7,44 (1 H, d, =CH-S) J CH=CH(Z) = 11 Hz; 9,52 (1H, s, -CONH).
EXAMPLE 9 To a solution containing the 7,3 -[(2 - cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - cephalos poranic acid (3.2 g) and NaHCO3 (0.63 g) in 100 ml of water, the sodium salt of 1 - methyl - 5 - mercapto - 1, 2,3,4 - tetrazole (1.55 g) is added and the mixture is heated, with stirring, for 6 hours at 60-65 C, main taining the pH in the range of 6.4-6.8. At the end of the reaction the mixture is cooled and washed with ethyl acetate; the aqueous phase is extracted with ethyl acetate and acidified with stirring with 40% H3PO4; the organic layer is separated, washed with water saturated with NaCI, dried and evaporated to dryness. The product thus obtained contains impurities which are not easily eliminated.The product can be purified by forming its benzhydryl ester: the raw product is dissolved in acetonitrile (50 ml) and to the solution there is added a solution containing diphenyldiazomethane (3.6 g) in acetonitrile (50 ml) and the mixture is stirred at room temperature for 1 hour. Few drops of acetic acid are then added and the mixture is evaporated, taken up with ethyl acetate, washed first with Na2HPO4 and then with water saturated with NaCI. The organic phase is separated, dried and evaporated to dryness under reduced pressure. The residue is purified through separation on a silica gel coumn, using as an eluent ethyl acetate : cyclohexane (7:3), thus obtaining the benzhydryl ester. The ester is hydrolyzed by treatment with trifluoroacetic acid (25 ml) and anisole (2.5 ml) at-i 5 C for 5 minutes.After evaporation under reduced pressure, the residue is redissolved in a small amount of ethyl acetate and precipitated with ethyl ether thus obtaining the 7,3 -[2 cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - [(1 methyl 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 cephem -4 - carboxylic acid, which results identical under l.R., U.V., microanalysis and N.M.R. to the product already described in Ex. 4 EXAMPLE 10 With a process similar to that described in Ex. 9 the following compounds were prepared: a) 7,32-cyano-vinylene (Z)-thioacetamido]-7amethoxy-3{(1 H-1,2,3,4-tetrazol-5-yl)- thiomethylt - 3 - cephem - 4 - carboxylic acid; b) 7ss{-2-cyano-vinylene (Z)-thioacemido]-7a- methoxy - 3 - -ethyl-1,2,3,4-tetrazol-5-yl)- thiomethyl] - 3 - cephem - 4 - carboxylic acid; c) 7,3 - [2- cyano - vinylene (Z) - thioacetamido] - 7a methoxy - 3- [1 - (2) - propenyl - 1,2,3,4 - tetrazol -5 - yI) - thiomethyl - 3 - cephem - 4 - carboxylic acid.
EXAMPLE 11 To a solution containing the 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - cephalosporanic acid (3,2 g) and Na H CO3 (0,63 g) in 100 ml of water there is added the disodium salt of 1,3 - carboxyethyl - 1,2,3,4 - tetrazole (2,4 g) and the mixture is heated, with stirring, for 6 hours at 65-70"C, maintaining the pH in the range of 6,4 - 6,8. At the end of the reaction the mixture is cooled and washed with ethyl acetate; the aqueous phase is extracted with ethyl acetate, acified, under stirring, with 40% H3PO4; the organic layer is separated, dried and evaporated to dryness and the product thus obtained is purified by forming its corresponding benzhydryl diester.To effect this purification, the raw product is dissolved in acetonitrile (80 ml) and to the solution there is added a solution containing diphelyldiazo - methane (7 g) in acetonitrile (70 ml).
The mixture is stirred at room temperature for 1 hour, and then few drops of glacial acetic acid are added. The mixture is then evaporated, the residue is taken up with ethyl acetate, washed first with Na2 HPO4 and then with water saturated with Na Cl. The organic phase is separated, dried and evaporated to dryness under reduced pressure.
The residue is purified by separation on a silica gel column, using as an eluent ethyl acetate: cyclohexane (7:3). The ester is hydrolized by treatment with trifluoroacetic acid (50 ml) and anisole (5 ml) at-15 C for 10 minutes. After evaporation under reduced pressure, the residue is redissolved in a small quantity of ethyl acetate and precipitated with ethyl ether, thus obtaining the 7,3 -[2 - cyano - vinylene (Z) thioacetamido[ - 7a - methoxy - 3 - [(1 -,3 - carboxyethyl - 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 cephem - 4 - carboxylic acid.
Elemental analysis (for Cr8HrgN707S3) Found: C: 39.71; H: 3.63; N: 17.90; S: 17.45 Calculated: C: 39.95; H: 3.54; N: 18.10; S: 17.77 I.R. (KBr): v(C=N) conjugated 2220 cm-' y(C=O) ss-lactam 1780 cm-' v(C=O) conjugated acid 1720 cm-' y(C-N) +8 (N-H) sec-amide 1540 cm-' U.V. (phosphate buffer, pH 7.4):#max = 275 mp E1%1Cm = 405 T.L.C.: Rf = 0.53 (CHCI3: MeOH:HCOOH = 8:2:1) N.M.R.: ppm (DMSO-d6) 2.93 (2H, t, -CH2 - CH2 - COOH on the tetrazole ring 3.47 (3H, s, 7-OCH3) 3.83 (4H, br-s, 2-OH2 and -S-CH2-CO)
5.00 (1H, s,6-H) 5.72 (1 H, d, NC-CH=) 7.66 (1 H, d, =CH-S) J CH=CH(Z) = 11 Hz 9.56 (1 H, s, -CONH).
Example 12 With a procedure similarto that described in Ex. 11, starting from the 7,3i2-cyano-vinylene (Z) thioacetamidoj - 7a - methoxy - cephalosporanic acid, the following compounds were prepared: a) 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] - 7a methoxy - 3- [(1 - carboxymethyl -1,2,3,4 - 3, tetrazol 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid.
Elemental analysis (for C,7H,7N707S3): Found: C: 38.55; H: 3.41; N: 18.40; S: 17.95 Calculated: C: 38.75; H: 3.25; N: 18.60; S: 18.15 I.R. (KBr): v(CsN) conjugated 2220 cm-' v(C=O) p-lactam 1780 cm-' v(C=O) conjugated acid 1720 cm-' v(C=N) +8 (N+H) sec-amide 1540 cm-' U.V. (phosphate buffer, pH 7.4): Xmax = 274 m,u E'%1cm = 410 T.L.C.: R, = 0.63 (CHCI3: MeOH:HCOOH = 8:3:1) N.M.R.: ppm (DMSO-d6) 3.47 (3H, s, 7-OCH3) 3.71 (4H, br-s, 2-CH2 and -S-CH2-CO) 4.33 (2H, q, 3-CH2) 5.07 (1 H, s,6-H)
5.72 (1H, d, NC-CH =) 7.66 (1 H, d, =CH-S) J c=cH(z) = 11 Hz 9.56(1H,s,-CONH); b) 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] - 7a methoxy - 3 -[(1 - - carboxypropyl - 1,2,3,4 tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - car boxylic acid;; c) 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] - 7a methoxy - 3 -[(1 - ca rboxyamidomethyl - 1,2,3,4 tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - car- boxylic acid; d) 7ss - [2 - cyano - vinylene (Z) - thioacetamide] 7a methoxy - 3 -[(1 -,3 - carboxyamidoethyl - 1,2,3,4 tetrazol - 5 - yl) -thiomethyl] - 3 - cephem - 4 - car- boxylic acid;; e) 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] - 7a methoxy - 3 -[(1 - 7- carboxyamidopropyl - 1,2,3,4 tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - car- boxylic acid.
Example 13 To a solution of the 2 - carboxyamido - vinylene (Z) mercaptoacetic acid (0.81 g) in acetonitrile: dimethylformamide (2:1; 60 ml), there is added triethylamine (0.7 ml) and 2 drops of N-methylmorpholine. The mixture is cooled to -50 C and dropwise with stirring, a solution of pivaloyl chloride (0.61 ml) in anhydrous acetonitrile (10 ml) is then added. The mixture is stirred for 30 minutes at +5 C and there is then added a solution containing the t-butyl ester of the 7,3 - amino - 7 [- methoxy - 3 [(1 - methyl - 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] 3 - cephem - 4 - carboxylic acid (2.25 g) dissolved in anhydrous sym-dichloroethane (40 ml).The reaction mixture is stirred for 1 hour at OOC and then for2 hours at room temperature; it is diluted with ethyl acetate, washed with a 3% KHSO4 solution and then with water. It is then washed with a solution of NaHOO3 and again with water. The organic phase is separated, dried over Na2SO4, filtered, and evaporated to dryness under reduced pressure; the residue thus obtained is purified on a silica gel column (150 g) using as an eluent a mixture of ethyl acetate : cyclohexane (3:1).The solution is evaporated to dryness, the residue is taken up with ethyl ether and is filtered thus obtaining, the t-butyl ester of the 7,3 - [2 - carboxyamido - vinylene (Z) thioacetamido] - 7a - methoxy - 3 -[(1 - methyl - 1,2, 3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 carboxylic acid. This ester is hydrolyzed with trifluoroacetic acid and anisole as described in Ex. 4, thus obtaining (60%) 7,3 -[2 - carboxyamido vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - [(1 - methyl 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem 4 - carboxylic acid.
U.V. (phosphate buffer, pH 7.4): AmaX = 273 m,a E'%,Cm = 405 T.L.C.: Rf = 0.48 (CHCl3: MeOH: HCOOH = 8:1:1) I.R. (KBr): v(C=O) ss-lactam 1780 cm- #(C=O) acid 1660 cm- v(C-N) +8 (N+H) sec-amide 1550 cm- N.M.R.: ppm (DMSO-d6) 3.36 (2H, s, -S-CH2-CO) 3.40 (3H, s, 7-OCH3) 3.60 (2H, q, 2-OH2)
4.36 (2H, q, 3-OH2) 5.00 (1 H, s, -6-H) 5.90 (1H, d, -CO-CH=) 6.87 (1 H, br-s, H2N-CO-) 7.05 (1 H, d, =CH-S) J CH=CH(Z) = 11 Hz 9.56(1H,s,-CONH) Following the procedure outlined above, there was prepared the following compound: 7,3 - [2 - carboxamido - vinylene (Z) - thioacetamido] 7a- methoxy-3 -C(1-ethyl-1,2, 3, 4-tetrazol-5-yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid.
The,3 - carboxamido - vinylene (Z) - thioacetic acid used as starting material was prepared as follows: to a solution of propiolamide (6.9 g) in water (20 ml) there was added, with stirring at a temperature of about 0 C, a solution of 70% thioglycolic acid (10 ml) in 20% NaOH (18.9 ml). The solution was then stirred for 1 hour at 0 C and then for 1 hour at room temper ature. After stirring the solution was acidified with a stoichiometric amount of 70% perchloric acid. After cooling to about 5-0 C, the solid precipitate is filtered, taken up in water (45 ml), stirred for 10 minutes, filtered again and dried, thus obtaining 12.6 g of a mixture (9:1) of ss - carboxamido - vinylene (Z) thioacetic acid.The two acids have the same solubil ity in water and therefore, by stirring the mixture with an appropriate amount of water, it is possible to dissolve the E-isomer, leaving still undissolved about 8/9 oftheZ-isomer[the purification process may be controlled by means of T.L.C. (ether: pet roleum ether: formic acid = 15.5 :1)],forexample, by means of three successive washings (two with 50 ml of water and then again with 100 ml of water) thus obtaining 10.9 g of the pure Z - isomer (yield 72%), m.p. 180-181 C.
Elemental analysis (for C > H7NO3S): Found: C: 37.22; H: 4.37; N: 8.66; S: 20.00 Calculated: C: 37.25; H: 4.37; N: 8.69; S: 19.89 I.R. (KBr): v(N-H) - NH2 group 3450,3210cm-1 v(C=O) acid 1685 cm-' v(C=O) amide 1625 cm-' N.M.R. (DMSO-d6): 3.43 (s,-S-CH2-) 5.94 (d, -CO-CH=) 6.97 (d, =CH-S) 7.16(d,-OONH2) 12.00 (br, -S-COOH) J CH=CH(Z) = 10 Hz.
Example t4 To a solution of 2 -t - butoxycarbonyl - vinylene (Z) mercaptoacetic acid (3.27 g) in anhydrous dichloroethane (100 ml) there is added triethylamine (2.11 ml) and two drops of N - methylmorpholine. The mixture is cooled to -5,0 and then there is added dropwise, with stirring, a solution of pivaloyl chloride (1.83 ml) in anhydrous dichlororoethane (20 ml). The mixture, is stirred for 30 minutes at +5 C, then a solution containing the t - butyl ester of the 7,3 - amino -7a- methoxy-3 -[(1 - (2) - propenyl - 1,2,3, 4. tetrazol - S - yI) - thiometyl] - 3 - caphem - 4 carboxylic acid (2.38 g) dissolved in anhydrous sym-dichloroethane (40ml) is added.The reaction mixture is stirred for 1 hour at 0 C, and then for 2 hours at room temperature, it is diluted with ethyl acetate, washed first with a 3% K H SO4 solution and then with water. The organic phase is separated, dried over Na2SO4, filtered and evaporated to dryness under reduced pressure. The resulting residue is purified on a silica gel column (150 g) with a mixture of ethyl acetate: cyclohexane (2:1). The solution is evaporated to dryness, taken up with ethyl ether and filtered, thus obtaining the t-butyl ester of the 7,3 i2-butoxy-carbonyl-vinylene (Z)-thioacetamido]7a- methoxy-3-t(1 - (2) - propenyl 1,2,3,4- tetrazol - 5 - yl) - thiomethyla - 3 - cephem - 4 - carboxylic acid.
By treatment with trifluoroacetic acid and anisole as described in Ex. 4, there is achieved the hydrolysis both of the ester on the carboxyl present in position 4 and on the carboxyl present on the acyl chain, thus obtaining the 7,3 - [2 - carboxy - vinylene (Z) thioacetamido] - 7a - methoxy - 3. [(1 - (2) - propenyl -1,2,3,4 - tetrazol -5 - yl) - thiomethyg - 3 - cephem 4- carboxylic acid.
U.V. (buffer phosphate, pH 7.4): AmaX = 273 m T.L.C.: R, = 0,54 (CH Cl3: OH3OH : HCOOH = 16:5: 2) I.R. (KBr): y(C=O) ss-lactam 1760cm-1 #(C=O) acid 1650 cm-' v(C-N) +8 (N-H) sec-amide 1575 cm-' N.M.R. ppm (DMSO-d6) 3.40 (3H, s, 7-OCH3) 3.42 (2H, s, -S-Cll2 - CO) 3.60 (2H, q, 2-OH2) 4.36 (2H, q, 3-OH2) 4,95 (2H, m, N-CH2-) 5,00 (1 H, s,6-H)
5,84(1 H, d, -CO-CH=)
7,05 (1H, d, =CH-S) 9,56 (1H, s,-CONH).
Operating in a similar manner the following compounds were obtained: a) 7,3 - [2 carboxy - vinylene (Z) - thiocetamido} - 7a methoxy-3-[(1 - methyl - 1,2,3,4-tetrazol-S-yl) - thiomethyq - 3 - cephem - 4 - carboxylic acid; b) 7,3 - [2 - carboxy - vinylene (Z) - thioacetamido] - 7a - methoxy-3-[(1 -ethyl- 1, 2, 3, 4 - tetrazol -S-yl) thiomethyl] - 3 - cephem - 4 - carboxylic acid.The 2 - t - butoxycarbonyl - vinylene (Z) mercaptoacetic acid used as starting material was prepared as follows: to a solution of the t - butyl ester of propiolic acid in acetone (126 ml) and water (63 ml), cooled to 0 C, there was added dropwise, with stirring, a solution containing 95% thioglycolic acid (7ml) in 2N NaOH (47.5 ml) and water (52.5 ml), keeping the temperature at about 000. At the end of the addition the mixture is stirred for 30 minutes at 0 C and for 1 hour at room temperature. The acetone is evaporated, the residue taken up with water, brought to an alkaline pH with some drops of dilute NaOH and washed with ethyl acetate. The extracts are washed with water, then with aqueous NaCI solution and dried over Na2SO4.After filtration and evaporation to dryness, there is obtained an oily residue which is purified by crystallization from C 014 (150 ml), obtaining 17 g (78%) of product in the form of white crystals, m. p.
102-105 C.
Found: C: 49.31; H: 6.44; S: 13,96 (For C5H,404S) Calculated: C: 49.52; H: 6.46; S: 14.68 cm-' I.R. (K Br): y(C=O) ester 1710 cm-1 v(C=O) acid 1690 cm~' v(C=C) cm-' v(C-C) t-butyl 1370 cm-' v(C-O-C) 1160 - 1150 cm- N.M.R. (CDCl3) 1.48 (9H, s, -CO-O-t.Bu) 3.42 (2H, s, -SCH2-COOH) 5.84 (1H, d, O-CO-CH=) 7.08 (1H, d, =CH-S-) JCH=CH(Z) = 10 Hz EXAMPLE 15 To a solution of the 2 - t - butoxy - carbonyl - vinylene (Z) - mercapto - acetic acid (9.05) in anhydrous ether (250ml) there is added at-10 C, in a single portion, 8.64 g of P-Cl5. The resulting suspension is stirred for 5 minutes at -10 C and then for 2 hours at 2-5 C. There is obtained a colorless solution which is evaporated under reduced pressure, taken up with anhydrous benzene and re-evaporated until all the PO 012 formed in the reaction is removed. There is obtained the 2 -t - butoxycarbonyl - vinylene (Z) mercaptoacetyl chloride in the form of a white crystalline mass.
In another vessel is prepared a solution of the benzhydryl ester of the 7,3 - amino 7a - methoxy cephalosporanic acid (15.6 g) in anhydrous sym dichloroethane. To this solution, cooled to --10 C, there is added N,N - diethylaniline (6.62 ml) and, immediately after, the aforementioned acid chloride, dissolved in sym-dichloroethane (50 ml). At the end of the addition, the mixture is stirred for 20 minutes at-5 C and then for 1 hour at room temperature.
The solvent is removed under reduced pressure, the residue is taken up with ethyl acetate and the solution is washed, in the following sequences, with aqueous solutions of K H SO4, Na CI, Na H CO3 and Na Cl. The organic phase, thus washed, is dried over Na2SO4, evaporated and the residue is chromatographed on a column (1.5 Kg Si 2; mobile phase = ethyl ether) The fractions containing the pure product are combined, concentrated until there is evident an incipient separation of an oil, and poured, with stirring, into 4 volumes of cyclohexane. There are thus obtained 17.19g of the benzhydryl ester of the 7,3 - [2 - t - butoxy - carbonyl - vinylene (Z) thioacetamido] - 7a - methoxy - 3- acetoxymethyl - 3 - cephem - 4 - carboxylic acid, as a pale yellow powder.
The benzhydryl ester thus obtained was added, portionwise, to a mixture of trifluoroacetic acid (150 ml) and anisole (25 ml), cooled to 0 C and well stirred. At the end of the addition, the reaction flask is closed and the mixture is stirred at 0 C for 30 minutes. The trifluoroacetic acid is then evaporated without heating and the residue is triturated with ethyl ether. There is obtained a powder which is filtered, washed with ether and immediately placed in a vacuum drier. There are thus-obtained 10.05 g of 7,3 - [2 - carboxy - vinylene (Z) - thioacetamido] 7a - methoxy - 3 - acetoxymethyl - 3 - cephem - 4 - car- boxylic acid, in the form of a powder.
m.p. 120-122"C (with dec.) Elemental analysis (for C16 H18 N2 Og S2): Found: C: 43.15; H: 4.16; N; 6.12; S: 14.21 Calculated/C: 43.04; H: 4.06; N: 6.27; S: 14.36 I.R. (K Br): v(C=O) p-lactam 1775 cm-' v(C=O) acetoxy 1730 cm-1 v(C=O) conjugated acid 1680 cm-' N.M.R.8, ppm (DMSO-d6) 2,02 (3H,s, -OCOCH3) 3;;41 (3H,s,7a-OCH3) 3,44(1 H,d,2-CH2-) 3,49 (2H,s, -S CH2 CO-) 3,65(1 H,d, 2-OH2-) 4,69(1 H,d, 3 CH2 OAC) 5,03 (1 H,d, 3 CH2 OAC) 5,12 (1H,s, 6-H 5,84 (1 H,d, =CH -COOH) JCH=CH(cis)= 10 Hz 9,58 (1 H, broad S, -CONH-) EXAMPLE 16 To a solution of Na CO3 (1.765 g) in water (40 ml) there is added the 7,3 - [2 - carboxy - vinylene (Z) thioacetamido] - 7a - methoxy - 3 - acetoxy methyl - 3 -cephem-4-carboxylic acid (3.12 g) and then the 6mercapto - 8 - amino - tetrazolo [1,5 b] pyridazine (1.175 g).When the solution is completed, the pH is corrected to 4.7 by the addition of traces of Na H CO, or K H SO4, depending on the necessity. The reaction vessel is then immersed into a thermostatic bath at 100-105"C. After 45 minutes the reaction is stopped by adding crushed ice to the mixture. The mixture is then salted with NaCI, extracted with ethyl acetate (150 ml), and acified with H2SO4. The organic phase is separated and the aqueous phase is extracted twice (2x50 ml).
The combined organic phases are concentrated to a small volume. On addition of ether, there are precipitated 1.95 g of raw material. For the purification, said material was triturated in acetonitrile (80 ml) and dioxane (20 ml), and treated with a solution of diphenyldiazomethane (4.04 g) in acetonitrile (60 ml).
The mixture is stirred for 30 minutes at room temperature and the excess of diphenyldiazomethane is decomposed with acetic acid. The mixture is evaporated to dryness under reduced pressure and the resulting raw product is purified chromatographically on a column (500 g Si O2; mobile phase = ethyl acetate: cycloexane,2:1), obtaining, after tritu ration with ether and filtration, 2.6 g of the dibenzhydryl ester. This product was then added, in small portions, to a solution of trifluoracetic acid (25 ml) and anisole (3 ml) at a temp erature of O"C. After stirring for 10 minutes, the trifluoroacetic acid was removed under reduced pressure and the residue was triturated with ethyl ether (60 ml), filtered and washed with ether.
There where thus obtained 1.35 g of pure 7,3 - [2 carboxy - vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - (8 - amino - tetrazolo [1, 5b] - pyridazin - m.p.: 180 C (with dec.) Elemental analysis (for C,8 H18 N8 07 S3): Found: C: 39.1; H: 3.29; N: 20.05; S: 17.23 Calculated:C: 38.98; H: 3.27; N: 20.2; S: 17.34 I.R. (K Br) v(C=O) ss-lactam 1780 cm-' v(C=O) amide and conjugated acid 1710-1680 cm-' v(C=O) acid 1630 cm-' y(C-N) and 8 (N-H) sec-amide 1570 cm-' NMR =8, ppm (DMSO-d6) 3,41 (3H, s., 7a-OCU3) 3,48 (1 H, d., 2-CH2-) 3,55 (2H, s., -S-CU2-CO-) 3,80 (1 H, d., 2-CU2-) 4,07 (1 H, d., 3-CU2-S-) 4,53 (1H, d., 3-CU2-S-) 5,14(1 H,s., 6-H) 5,85(1 H, d., =CH-COO) 6,35 (1 H, s,7-H in the pyri Anzine ring) JCH=CH (cis) = 10 Hz 7,4(1 H, d., -CH=S-) 7,93 (2H, broad s., 8-NH2 in the pyridazine ring) G, HF (1 H, broad -S-CO-NH-) EXAMPLE 17 To a solution of the 7,3 - (bromoacetamido) - 7a methoxy-3-[(1 - methyl - 1,2,3,4-tetrazol -S-yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid (4.79 g) in 0.1 N sodium hydroxyde (100 ml) there is added the ss-mercapto-acrylonitrile (Z) (0.9 g) dissolved in 105 ml of 0.1 N sodium hydroxide. The mixture is stirred for 2 hours at room temperature and is then extracted with ethyl acetate (200 ml) and, with stir ring, is acidified with 6 N H Cl at 0-5 C to pH2.
The organic phase is separated and the water phase, after saturation with NaCI, is extracted three more times. The organic extracts are combined, dried over sodium sulfate and evaporated to a small volume. After addition of ethyl etherthere are obtained about4 g of -[2 - cyano - vinylene (Z) thioacetamido] - 7a - methoxy - 3 -[(1 - methyl - 1,2, 3,4 tetrazol - 5 - yl) - thiomethyl] - 3 - cephenl - 4 carboxylic acid, which results identical under I.R., U.V. and N.M.R. to the product already described in Ex. 4. With the same procedure there were also obtained the compounds reported in the preceding examples.
EXAMPLE 18 To a solution of 7 - (mercapto - acetamide) - 7a methoxy -3 -[(i - methyl t,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephenl - 4 - carboxylic acid (4.729) in 0.1 N NaOH (4.32 g) there is added with stirring, a solution of,3- chloroacrylonitrile (Z) (0.87 g) in 0.1 N NaOH (100 ml).
The mixture is stirred for4 hours at room tempera ture, is then extracted with ethyl acetate (200 ml) and, with stirring, is acidified with 6N H Cl at 0-5"C to pH2. The organic phase is separated and the aque ous phase, saturated with NaCI is extracted three more times with fresh ethyl acetate. The organic extracts are combinated, dried over sodium sulfate, concentrated to a small volume and poured, with stirring, into ethyl ether. There is thus obtained the 7,3 - [2- cyano - vinylene (Z) - thioacetamido] - 7a methoxy-3{(i-methyl-1 ,2,3,4-tetrazol-5-yl) thiomethyl]-3-cephemA-carboxylic acid (4.729) results identical under l.R., U.V. and N.M.R. to the compound already described in Ex. 4.
With the same procedure there were also obtained the compounds described in the preceding examples.
EXAMPLE 19 To 150 ml of anhydrous tetrahydrofuran there is first added 22 ml of a 2 M solution of lithium methylate in methanol and then 35 ml of methanol. After 10 minutes, the mixture is cooled to -70 C and then there is added thereto the benzhydrylic ester of the 7,3 -[2 - cyano - vinylene (Z) - thioacetamide] - 3 -[(1 methyl-1,2,3,4-tetrazol-5-yI)-thiomethyl] - 3 - cephem - 4 - carboxylic acid (7.25) dissolved in anhydrous tetrahydrofuran (50 ml) and the reaction temperature is maintained at-65 C. After 5 minutes there is added rapidly 1.26 ml of t-butyl hypochlorite and the mixture is stirred for 40 minutes.There are then added 28 ml of acetic acid and then trimethylphosphite (0.5 g) to destroy the excess of oxidizing agent The mixture is then evaporated under reduced pressure, the residue is taken up with methylene chloride, washed first with water and then with a solution of NaHOO3, dried and evaporated under reduced pressure thus obtaining (in 50% yie!d) the benzhydrylic ester of the 7,3 -[2 - cyano vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - [(1 methyl-1,2,3,4-tetrazol-S-yI)-thiomethyl] - 3 - cephem - 4 - carboxylic acid.Said ester is hydrolyzed by treating it with trifluoroacetic acid thus obtaining the 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] - 7a -methoxy-3-[(1 methyl 1, 2, 3, 4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid.The benzhydrylic ester of the 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] - 3 - [(1 - methyl - 1,2,3,4 - tetrazol 5 - yl) - thiomethyl] - 3 - cephen - 4 - carboxylic acid, used as starting material, was prepared according to the following method: To a solution of 7,3 -[2 - cyano - vinylene (Z) thioacetamido] - 3 - [(1 - methyl - 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid (8.94 g) in anhydrous acetonitrile (70 ml) there is added dropwise, at room temperature and with stirring, a solution of diphenyldiazomethane (3.89 g) in anhydrous acetonitrile (70 ml).After 30 minutes from the end of the addition there is added 1 ml of acetic acid to decompose the eventually unreacted diphenyldiazomethane. The acetonitrile is evaporated under reduced pressure, without heating, and the resulting residue is dissolved in a minimum amount of acetone and precipitated, with vigorous stirring, with n-hexane. There is obtained a paste-like product which solidifies on trituration. The solid, well triturated, is poured into 50 ml of ethyl ether and stirred for 30 minutes. The powder, filtered and washed with additional ethyl ether, weighs 11.5 (yield 94.4%) and consists of the benzhydrylic ester of good purity.
An additional purification may be effected, if it is so desired, by passing through an Si O2 column (mobile phase=ethyl acetate: cyclohexane, 3:1) and/or by recrystallization from benzene (about 50 ml per gram). The pure product melts at 113-114 C (with dec.).
EXAMPLE 20 To a solution containing the t-butyl ester of the 7 (benzylidene - amino) - 3 -[(1 - methyl - 1,2,3,4 tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid (4.72 g) in anhydrous dimethoxyethane (50 ml), cooled to -5 C, there is added, under a nitrogen stream, potassium t - butoxide (1.12 g). The solution is stirred for 1 minute and there is then added methyl - methanethiolsulphonate (1,26 g) dissolved in dimethoxyethane (2 ml). The solution is stirred for 5 minutes and then poured into a solution of phosphate buffer at a pH of 6.6.The mixture is then extracted with CH Cl3, the organic phase is washed with a saturated saline solution, dried over sodium sulfate and evaporated thus obtaining (yield 50 /O) the t - butyl ester of the 7,3 - (benzylidene amino) - 7a - methylthio -3 -[(1 - methyl - 1,2,3,4- tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - car- boxylic acid, which is purified chromatographically on a column of Si O3 (completely anhydrous) using as a mobile phase an ethyl ether; petroleum ether mixture (3:1).To a solution of this compound (5.18 g) in 100 ml of CH2CI2 there is added, under nitrogen and at room temperature, the 2 - cyano - vinylene (Z) - thioacetyl chloride (1.62 g). There is then added dropwise 0.2 ml of water and the mixture is stirred overnight. At the end the organic layer is separated, washed with water, dried over sodium sulfate and evaporated thus obtaining the t-butyl ester of the 7,3 - [2 - cyano - vinylene (Z) - thioacetamido] - 7 - methylthio-3-[(1 methyl-l, 2, 3, 4-tetrazol-5-yl) - thiomethyl - 3 - cephem - 4 - carboxylic acid, which is purified chromatographically on an Si O2 column, using as an eluent a mixture of ethyl acetate: cyc lohexane (3:1).A solution of said purified ester (2.78 g) in methanol (25 ml) is brought to refluxtemperature, under a nitrogen atmosphere. There isthen added mercuric acetate (1.6 g) and the mixture stir red under reflux for an additional ten minutes. The mixture is cooled, taken up with benzene-water and the organic phase, after repeated water washings, is dried on sodium sulfate and evaporated to give an oil, which consists of the raw t - butyl ester of the 7ss - [2 - cyano - vinylene (Z) - thioacetamido] - 7a - methoxy-3-[(1 - methyl - 3, 4-tetrazol-5-yl)- thiomethyl] - 3 - cephem - 4 - carboxylic acid. The oil thus obtained is purified on a silica gel column following the procedure described in Ex. 3, obtaining a compound identical to that described in said EX 3.
The hydrolysis of said ester was carried out as described in Ex. 4, obtaining a compound identical to that obtained in Ex 4.
EXAMPLE2 1 To a solution of 10.28 of bis - (trimethylsilyl) - trifluoroacetamide in 100 ml of anhydrous CH2CI2 there is added the 2 - cyano - vinylene (Z) - thioacetyl chloride (6.46 g) and the mixture is stirred for 30 minutes at room temperature. To this solution there is added 9.34 of the dibenzhydryl ester of the 7,3 - (D 5 - t - butoxycarbonylamino - 5 - carboxyvaleryl) - 7a -methoxy-3-[(1 methyl - 1,2,3,4-tetrazol -S-v) - thiomethyl] - 3 - cephem - 4 - carboxylic acid (prepared according to Chem. Pharm. Bu11,24, (11), 2629 (1976) ). This solution is refluxed for 70 hours, with stirring, cooled, poured into an aqueous icy solution of bicarbonate and stirred for 30 minutes.The methylene chloride phase is separated and the water phase is extracted with CHCl3. The combined organic phases are washed with a saturated aqueous solution of Na CI, dried over sodium sulfate and evaporated under reduced pressure. There is thus obtained a residue consisting of the raw benzhydrylic diester of the 7ss - [N - (D - 5 - t - butoxycarbonylamino - 5' - carboxyvaleryl) - 2 - cyano - vinylene (Z)-thioacetamido]-7a-methoxy-3i(l- methyl - 1,2,3,4 - tetrazol - 5 - yl) - thiomethylt - 3 - cephem - 4 - carboxylic acid, which is then treated without any additional purification, with trifluoroacetic acid (30 ml) and anisole (10 ml) for 10 minutes at 000. The mixture is then evaporated to dryness under reduced pressure and the residue is dissolved in a 10% aqueous K2H PO4 solution and ethyl acetate.
The aqueous solution is separated, washed further with ethyl acetate, layered with ethyl acetate and acified with K2PO4 to pH 2.5. The organic phase is separated and on the aqueous phase, after a salting out operation, there are effected two additional ethyl acetate extractions. After drying over sodium sulfate, the evaporation of the solvent leaves a residue consisting of about 3.5 g of the 7,3 - cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - [(1 - methyl 1,2,3,4 - tetrazol - 5 - yl) -thiomethyl] - 3 - cephem 4 - carboxylic acid, which results identical under l.R., U.V. and N.M.R. to the compound already described in Ex.4.
EXAMPLE 22 To, a suspension of the 7,3 -[2 - cyano - vinylene (Z)-thioacetamido] -7a-methoxy-3i(1 -methyl-1, 2, 3, - tetrazol - 5 - yl) - thiomethylt - 3 - cephem - 4 - carboxylic acid (5.60 g) in water (80 ml), there is added a stoichiometric quantity of sodium bicarbonate, obtaining the complete solution of the compound. The solution is then lyophilized, obtaining the sodium salt of the 7,3 - [2 - cyano - vinylene (Z) thioacetamido] - 7a - methoxy - 3 - [(1 - methyl - 1, 2, 3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 carboxylic acid.By carrying out in a similar manner the operation described above, there are obtained the sodium salts of the compounds described in the preceding examples.
EXAMPLE 23 To a solution containing the 7,3 -[2 - cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - (1 methyl - 1,2,3,4 - tetrazol - 5 - yl) -thiomethyl - 3 - cephem - 4 - carboxylic acid (5.60 g) in ethyl acetate (120 ml), there is added a stoichiometric quantity of a 30% solution of sodium 2 - ethyl - hexanoate in isopropyl alcohol.After stirring for 30 minutes at room temperature, the mixture is diluted with petroleum ether and the resulting precipitate is removed by filtration, thus obtaining the sodium salt of the 7,3 - [ 2cyano - vinylene (Z) - thioacetamido] - 7a - methoxy 3 - [(1 -methyl-1,2,3,4-tetrazol-S-yl)- thiomethyl] - 3 - cephen -4 - carboxylic acid. By utilizing a similar procedure, there are obtained the sodium salts of the compounds described in the preceding examples.
EXAMPLE24 To a solution of the sodium salt of the 7,3 - [2 cyano - vinylene (Z) - thioacetamido] - 7a - methoxy 3 - [(1 -methyl-1,2,3,4-tetrazol-5-yI)- thiomethyl] - 3 - cephem -4 - carboxylic acid (6.39 g) in acetone (80 ml) there is added a solution of 10% sodium iodide in water (0.8 ml) and chloromethyl pivalate (1.44 ml). The suspension is then refluxed for 3 hours and then cooled to 5 C. The resulting solid is removed by filtration and the filtrate is evaporated under reduced pressure. The oily residue is dissolved in ethyl acetate (100 ml) and the resulting solution is washed with a 5% aqueous sodium bicarbonate solution and then again with water.The solution is dried and evaporated to dryness under reduced pressure, thus obtaining the pivaloyloxymethyl ester of the 7,3 - [2 - cyano - vinylene (Z) thioacetamido] - 7a - methoxy - 3 - [(1 - methyl - 1, 2, 3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem -4 - carboxylic acid.
By following a similar procedure, the pivoloyloxymethyl esters of the compounds illustrated in the preceding examples were prepared.
EXAMPLE25 An injectable pharmaceutical composition was obtained by dissolving 100-500 mg of the sodium salt of the 7,3 - 2 - cyano - vinylene (Z) thioacetamido - 7a - methoxy - 3 - [(1 - methyl - 1,2, 3,4 - tetrazol - 5 - yl) - thiomethyl - 3 - cephem - 4 carboxylic acid in sterile water or in a sterile saline solution (1-2 ml).

Claims (8)

1. A compound having the general formula (I)
wherein Z is cyano, carbamoyl, -COOR wherein R is hydrogen or C,-C6 alkyl; X is a free or esterified carboxy group; Airs: a)
wherein Y is a') hydrogen; b') C1-C6 alkyl; c') C2-C6 alkenyl; d') -(CH2),-CN wherein n is an integer of 1 to 4; e') -(CH2)n-COOR, wherein n and R are as defined above; f') -(OH2),-CONH2, wherein n is as defined above; or b)
wherein R, is a") hydrogen;
wherein each of the groups R, which may be the same or different, is as defined above; d") -NHCONH2; e") -NHCH2COOR; f") -COOR; g") -CH = CH-COOR; h")-(CH2)m-COOR; i")-(CH2)m-CN; wherein R is as defined above and m is zero or an integer of 1 to 3, and the pharmaceutically or veterinarily acceptable salts thereof.
2. A compound having the formula (I) above reported in claim 1, wherein Z is cyano, carbamoyl or -COOR wherein R is hydrogen or C,-C6 alkyl; X is a free or salified carboxy group and A is: a)
wherein Y is -CH3; - (OH2), - CN; (OH2), - COOH or - (OH2), - CONH2, wherein n is an integer of 1 to 4; or b) wherein R, is hydrogen or amino, and the pharmaceutically or veterinarily acceptable salts thereof.
3. The cis-isomer of a compound of claim 2.
4. A compound selected from the group consisting of: 7ss-[2- cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - [(1 H - 1,2,3,4 - tetrazol - 5 yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7,3 - cyano - vinylene (Z) - thioacetamido] - 7&alpha;- methoxy- 3 -[(i - methyl - 1,2,3,4 -tetrazol - S - yl) thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7ss-[2- cyano-vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - [(1 -ethyl-1,2,3,4-tetrazol-5-yl)- thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7ss-[2 - cyano - vinylene (Z) - thioacetamido] - 7a - methoxy-3-[(1 - carboxy - methyl - 1,2,3,4 tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - car- boxylic acid; 7,3 - [2- cyano - vinylene (Z) - thioacetamido] - 7a methoxy-3-[(1 - ss - carboxy ethyl - 1,2,3,4- tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - car- boxylic acid; 7ss-[2- cyano - vinylene (Z) - thioacetamido] - 7a - methoxy-3-[(1 -y-carboxypropyl - 1,2,3,4 tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - car- boxylic acid; 7ss-[2- cyano - vinylene (Z) - thioacetamido] - 7a - methoxy-3-[(1 -(2)-propenyl-1,2,3,4-tetrazol-5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7,3 - [2- cyano - vinylene (Z) - thioacetamido] - 7a methoxy - 3 -[(1 - carboxamidomethyl- 1,2,3,4- tetrazol - 5 - yl) - thiomethyl] - 3- cephem - 4 - car- boxylic acid; 7,3 - cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3 - [(1 -,3 - carboxamidoethyl - 1, 2, 3, 4 tetrazol - 5 - yl) - thiomethyl3 - 3 - cephem - 4 - car- boxylic acid;; 7,3 - [2- cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3 -[(1 - - carboxamidopropyl - 1, 2, 3, 4 tetrazol-5-yl)-thiomethyl]-3-cephem-carboxylic acid; 7ss-[2- cyano - vinylene (Z) - thioacetamido] 7a methoxy-3 -[(1 -P- cyano ethyl -1,2, 3, 4-tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7ss-[2- cyano - vinylene (Z) - thioacetamido] - 7&alpha;- methoxy-3-[(1 - &gamma; - cyanopropyl-1,2,3,4-tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7ss-[2- carboxamido - vinylene (Z) - thioacetamido] - 7&alpha;- methoxy - 3 - [(1 - methyl - 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7ss-[2- carboxamido - vinylene (Z) - thioacetamido] - 7a!- methoxy-3-[(l-ethyl -1, 2, 3, 4-tetrazol-5-yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7ss -[2-carboxy- vinylene (Z) - thioacetamido] - 7amethoxy-3-[(1 methyl-1,2,3,4-tetrazol-5-yl)- thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7,3 - [2 - carboxy - vinylene (Z) - thioacetamido] - 7a methoxy-3-[(1 - ethyl 1, 2,3,4-tetrazol-5- yl)- thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7,3 - [2 - carboxy - vinylene (Z) - thioacetamido] - 7a methoxy- 3 - [(1 - (2)- propenyl - 1,2,3,4 - tetrazol - 5 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7ss-[2- cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3-[(tetrazolo [1 ,5-b] pyridazin - 6 - yl) thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7,3 - [2- carboxamido - vinylene (Z) - thioacetamido] - 7a- methoxy - 3-[(tetrazolo [1 ,5-b] pyridazin - 6 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7,3 - [2 - carboxy - vinylene (Z) - thioacetamido] - 7ametoxy - 3- [(tetrazolo [1,5-b] pyridazin - 6 - yI) - thiomethyl] - 3 - cephem - 4 - carboxilic acid; 7ss-[2- cyano - vinylene (Z) - thioacetamido] - 7a - methoxy - 3- [(tetrazolo [1,5-b] pyridazin - 8 - amino 6 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid; 7,3 - [2- carboxamido - vinylene (Z) - thioacetamido] - 7a- methoxy - 3-[(tetrazolo [1 ,5-b] pyridazin - 8 - amino - 6 - yl) thiomethyl] - 3- cephem - 4 - carbox- ylic acid; 7ss-[2- carboxy - vinylene (Z) - thioacetamido] - 7a - methoxy - 3- [(tetrazolo [1,5-b] pyridazin - 8 - amino 6 - yl) - thiomethyl] - 3 - cephem - 4 - carboxylic acid, and the pharmaceutically or veterinarily acceptable salts thereof.
5. A process for the preparation of a compound according to anyone of the preceding claims, the process comprising: A) reacting a compound of formula (II)
wherein Z and A are as defined above in claim 1 and X' is a free or salified or esterified carboxy group, with a compound of formula M-OCH2 wherein M is an alkaline metal in the presence of a positive halogenating agent; or B) reacting a compound of formula (III)
wherein Z is as defined in claim 1 and X' is as defined above, with a compound of formula (IV) HS-A (IV) wherein A is defined above in claim 1, our a reactive derivative thereof; or C) reacting a compound of formula (V)
Wherein X' is as defined above and A is as defined above in claim 1 and E is amino or a group -N=C= wherein O is an oxygen or sulphur atom, or a reactive derivative thereof, with an acid of formula (VI) Z-CH=CH-S-CH2-COOH (Vl) wherein Z is as defined above in claim 1, or a reactive derivative thereof; or D) reacting a compound of formula (VII)
wherein X' is as defined above, A is as defined above in claim 1 and Y is halogen, with a compound of formula (VIII) Z-CH=CH-SH (veil) wherein Z is as defined above in claim 1, or a reactive derivative thereof; or E) reacting a compound of formula (IX)
wherein X' is as defined above and A is as defined above in claim 1, or a reactive derivative thereof, with a compound of formula (X) Z-CH=CH-Y' (X) wherein Z is as defined above in claim 1 and Y' is halogen or the residue of a reactive ester of an alcohol; or F) reacting a compound of formula (Xl)
wherein X' is as defined above and Z and A are as defined above in claim 1, with methyl alcohol in the presence of Hg++ orAg+ ions; or G) removing the acylating aminoadipoyl group in a compound of formula (XII)
wherein Z, A and the group X', independently, are as defined above and T is an amino protecting group and, if desired, converting a compound of formula (I) into a pharmaceutically or veterinarily acceptable salt thereof and/or, if desired, obtaining a free compound from a salt and/or, if desired, converting a compound of formula (I), or a salt thereof, into another compound of formula (I), or a salt thereof, and/or, if desired, resolving a mixture of isomers into the single isomers.
6. A process according to claim 5 substantially as described in any one of the foregoing Examples.
7. A compound of formula (I) as defined in claim 1 where prepared by a process according to claim 5 or6.
8. A pharmaceutical or veterinary composition comprising a compound as claimed in anyone of the claims 1 to 4 and 7, and a pharmaceutically or veterinarily acceptable carrier and/or diluent.
GB7910345A 1979-03-23 1979-03-23 Unsaturated 3-heterocyclylthiomethyl-7a-methoxy-7-acylamido-3-cephem-4-carboxylic acid derivatives Expired GB2045233B (en)

Priority Applications (4)

Application Number Priority Date Filing Date Title
GB7910345A GB2045233B (en) 1979-03-23 1979-03-23 Unsaturated 3-heterocyclylthiomethyl-7a-methoxy-7-acylamido-3-cephem-4-carboxylic acid derivatives
DE19803006916 DE3006916A1 (en) 1979-03-23 1980-02-23 UNSATURATED 3-HETEROCYCLIC-THIOMETHYL-7 ALPHA -METHOXY-7 BETA -ACYLAMIDO-3-CEPHEM-4-CARBONIC ACID DERIVATIVES, METHOD FOR THE PRODUCTION THEREOF AND THEIR CONTAINING PHARMACEUTICAL AND VETERINARY MEDICINE
BE0/199885A BE882360A (en) 1979-03-23 1980-03-21 DERIVATIVES OF 3-HETEROCYCLO-THIOMETHYL-7ALPHA-METHOXY-7BETA-ACYLAMINO-3-CEPHEM-4-CARBOXYLIC ACIDS UNSATURATED
JP3580280A JPS55129291A (en) 1979-03-23 1980-03-22 Unsaturated 33heterocyclic thiomethyll 7 alphaamethoxyy7 betaaacylamidoo33cephemm 44carboxylic acid derivative

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GB7910345A GB2045233B (en) 1979-03-23 1979-03-23 Unsaturated 3-heterocyclylthiomethyl-7a-methoxy-7-acylamido-3-cephem-4-carboxylic acid derivatives

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0107928A1 (en) * 1982-09-30 1984-05-09 Shionogi & Co., Ltd. Vinylthioacetamido oxacephalosporin derivatives, and intermediates therefor, their preparation, formulations containing the same and their antibacterial use
FR2550200A1 (en) * 1983-08-01 1985-02-08 Fujisawa Pharmaceutical Co PROCESS FOR THE PREPARATION OF CEPHEM COMPOUNDS WITH ANTIMICROBIAL ACTIVITY AND NEW PRODUCTS THUS OBTAINED
CN102875577A (en) * 2011-07-14 2013-01-16 山东睿鹰先锋制药有限公司 Cefminox impurity D and preparation method thereof

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB8329030D0 (en) * 1983-10-31 1983-11-30 Fujisawa Pharmaceutical Co Cephem compounds
GB8406231D0 (en) * 1984-03-09 1984-04-11 Fujisawa Pharmaceutical Co Cephem compounds

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0107928A1 (en) * 1982-09-30 1984-05-09 Shionogi & Co., Ltd. Vinylthioacetamido oxacephalosporin derivatives, and intermediates therefor, their preparation, formulations containing the same and their antibacterial use
FR2550200A1 (en) * 1983-08-01 1985-02-08 Fujisawa Pharmaceutical Co PROCESS FOR THE PREPARATION OF CEPHEM COMPOUNDS WITH ANTIMICROBIAL ACTIVITY AND NEW PRODUCTS THUS OBTAINED
EP0133678A2 (en) * 1983-08-01 1985-03-06 Fujisawa Pharmaceutical Co., Ltd. New cephem compounds, processes for preparation thereof and pharmaceutical compositions containing them
EP0133678A3 (en) * 1983-08-01 1986-03-19 Fujisawa Pharmaceutical Co., Ltd. New cephem compounds, processes for preparation thereof and pharmaceutical compositions containing them
CN102875577A (en) * 2011-07-14 2013-01-16 山东睿鹰先锋制药有限公司 Cefminox impurity D and preparation method thereof
CN102875577B (en) * 2011-07-14 2014-07-02 山东睿鹰先锋制药有限公司 Cefminox impurity D and preparation method thereof

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DE3006916A1 (en) 1980-10-09
BE882360A (en) 1980-09-22
JPS55129291A (en) 1980-10-06

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