Cephalosporin intermediates

This invention relates to cephalosporin derivatives which have antibacterial activity.
According to the invention there is provided a cephalosporin derivative of the formula I:
[Formula I--given hereafter]
in which X is sulphur, oxygen, methylene or sulphinyl (R or S configuration);
R1 is 2-aminothiazol-4-yl or 2-aminooxazol-4-yl each optionally substituted in the 5-position by fluorine, chlorine or bromine, or R1 is 5-aminoisothiazol-3-yl, 5-amino-1,2,4-thiadiazol-3-yl, 3-aminopyrazol-5-yl, 3-aminopyrazol-4-yl, 2-aminopyrimidin-5-yl, 2-aminopyrid-6-yl, 4-aminopyrimidin-2-yl, 2-amino- 3,4-thiadiazol-5-yl or 5-amino-l-methyl-1,2,4-triazol-3-yl;
R50 is chloromethylene or a radical of the formula .dbd.N.O.R2, wherein R2 is hydrogen, (1-6C)alkyl, (3-8C)cyclo-alkyl, (1-3C)alkyl(3-6C)cycloalkyl, (3-6C)cycloalkyl(1-3C)alkyl, (3-6C)alkenyl, optionally substituted by carboxy, (5-8C)cycloalkenyl, (3-6C)alkynyl, (2-5C)alkylcarbamoyl, phenylcarbamoyl, benzylcarbamoyl, (1-4C)alkylcarbamoyl(1-4C)alkyl, di(1-4C)alkylcarbamoyl(1-4C)alkyl, (1-4C)haloalkylcarbamoyl(1-4C)alkyl, triphenylmethyl, (1-3C)haloalkyl, (2-6C)hydroxyalkyl, (1-4C)alkoxy(2-4C)alkyl, (1-4C)alkylthio(2-4C)alkyl, (1-4C)alkanesulphinyl(1-4C)alkyl, (1-4C)alkane-sulphonyl(1-4C)alkyl, (2-6C)aminoalkyl, (1-4C)alkyl-amino(1-6C)alkyl, (2-8C)dialkylamino(2-6C)alkyl, (1-5C)cyanoalkyl, 3-amino-3-carboxypropyl, 2-(amidinothio)ethyl, 2-(N-aminoamidinothio)ethyl, tetrahydropyran-2-yl, thietan-3-yl, 2-oxopyrrolidinyl, or 2-oxotetrahydrofuran-3-yl,
or --R2 is the formula --(CH.sub.2).sub.n --R6 in which n is 1 to 4 and R6 is piperidino, pyrrolidino, morpholino, piperazino or N-methylpiperazino, each value of R6 being optionally substituted by (1-4C)alkyl, phenyl or benzyl,
or --R2 is of the formula --(CH.sub.2).sub.m --W--R7 in which m is 0 to 3, W is sulphur or a direct bond, and R7 is phenyl or pyridinio(1-4C)alkylene or R7 is pyridyl, imidazolyl, 1,3,4-thiadiazolyl, tetrazolyl, 1-(1-4C)alkyltetrazolyl, thiazolyl, isothiazolyl or isoxazolyl in which the link with W is via a carbon or uncharged nitrogen, each value of R7 being optionally substituted, where possible, by one or two groups selected from (1-4C)alkyl, amino, hydroxy, carboxy, carbamoyl, nitro, (2-5C)alkoxycarbonyl, cyano or sulpho,
or --R2 is of the formula --(CH.sub.2).sub.n --CO--R8 in which n is 1 to 4 and R8 is (1-4C)alkyl, phenyl or benzyl,
or --R2 is of the formula --COR9 or --(CH.sub.2).sub.n --OCO--R9 in which n is 1-4 and R9is hydrogen, (1-4C)alkyl, (1-4C)haloalkyl, phenyl or benzyl,
or --R2 is of the formula --G--CH.sub.2 --R10 in which G is carbonyl or a direct bond and R10 is phthalimido,
or --R2 is of the formula --NR11R12R13 in which R11, R12 and R13 are (1-4C)alkyl, or R11 is (1-4C)-alkyl and R12 and R13 are joined to form a (3-6C)carbocyclic ring, or R11, R12 and R13 are joined to form a 1-azonia-4-azabicyclo[2,2,2]octane or 1-azonia-3,5,7-triazatricyclo[3,3,1,1.sup.3,7 ]decane,
or --R2 is of the formula II
[Formula II]
in which p is I or 2 and R14 and R15 are hydrogen or (1-4C)alkyl,
or --R2 is of the formula --P(O)R16R17 in which R16 is hydroxy, (1-4C)alkoxy, (2-8C)dialkylamino, phenoxy, phenylamino or one of the values given above for R6, and R17 is (1-4C)alkyl, (1-4C)alkoxy (2-8C)-dialkylamino, phenoxy, phenylamino, piperidino, pyrrolidino, morpholino, piperazino or N-methylpiperazino,
or --R2 is of the formula --CH.sub.2 P(O)R18R19 in which R18 and R19 are hydroxy or (1-4C)alkoxy,
or --R2 is of the formula --CH(SR20)COOR21 in which R20 is (1-4C)alkyl and R21 is hydrogen or (1-6C)alkyl,
or --R2 is of the formula III:
[Formula III]
in which m is 0-3, R22 is hydrogen, (1-3C)alkyl or methylthio, R23 is hydrogen, (1-3C)alkyl, (C3-C7)cycloalkyl, cyano, carboxy, (2-5C)carboxyalkyl or methanesulphonylamino, or phenyl optionally substituted by amino or hydroxy, or R22 and R23 are joined to form, together with the carbon to which they are attached, a (3-7C) carbocyclic ring, and R24 is hydroxy, amino, (1-4C)alkoxy, (1-4C)alkylamino, phenylamino or of the formula R6 given above or of the formula NHOR25 in which R26 is hydrogen, (1-4C)alkyl, phenyl or benzyl, provided that when R2 contains phenyl, and unless otherwise stated, the phenyl is optionally substituted by 1 or 2 groups selected from halogen, hydroxy, amino, carboxy, nitro, carbamoyl, cyano and aminomethyl;
R3 is hydrogen or methoxy;
R4 is hydrogen, (1-4C)alkyl, halo(1-4C)alkyl, hydroxy(1-4C)alkyl, (1-4C)alkoxy(1-4C)alkyl, carboxy (1-4)alkyl, amino (1-4C)alkyl, cyano(1-4C)alkyl, (1-4C)alkanoylamino(1-4C)alkyl, allyl, furfuryl, benzyl or pyridyl(1-4C)alkyl;
R5 is an aromatic heterocyclic ring system which is linked via carbon and is one of the formula IV to LI inclusive each of these ring systems being optionally substituted where possible, on a carbon atom or atoms, by one, two or three substituents selected from halogen, (1-6C)alkyl, carboxy, (2-6C)alkoxycarbonyl, (2-6C)- alkoxycarbonyl(1-4C)alkyl, (1-6C)alkoxy, (1-6C)alkylthio, cyano, (2-4C)cyanoalkyl, amino, (1-6C)alkylamino, (2 8C)dialkylamino, benzylamino (optionally substituted in the benzene ring thereof by nitro), thenylamino, allylamino, (1-6C)aminoalkylamino, (1-6C)alkoxy(1-6C)alkylamino, (1-6C)hydroxyalkylamino, hydroxy, mercapto, carbamoyl, (2-6C)alkylcarbamoyl, (3-10C)dialkylcarbamoyl, phenylthio and heteroarylthio wherein heteroaryl is a 5- or 6-membered ring containing 1, 2 or 3 hetero atoms selected from oxygen, nitrogen and sulphur;
and in which Y is oxygen, sulphur or NR27;
Z is nitrogen or CH;
one of A, B, D and E is +NR27 and the remainder are nitrogen;
and ring systems of Formula IV, XVI or XVII, which are optionally fused, on a carbon-carbon bond, with a 5- to 7-membered saturated carbocyclic ring;
R27 is nitrogen-linked and is (1-6C)alkyl, (1-6C)alkyl(2-6C)alkenyl, (2-6C)alkenyl, (2-8C)alkoxyalkyl, carboxy(1-6C)alkyl, [(1-6C)alkoxy]carbonyl(1-6C)alkyl, carbamoyl(1-6C)alkyl, carboxyamino-carbonyl(1-6C)alkyl, [(1-6C)alkoxy]carbonylamino-carbonyl(1-6C)alkyl, [(2-8C)alkanoyl]methyl, benzoylmethyl, (1-6C)hydroxyalkyl, (1-6C)alkylamino, or phenyl(1-6C)alkyl or phenyl, each optionally substituted by 1 or 2 groups selected from halogen, hydroxy, amino, carboxy, nitro, carbamoyl, cyano, trifluoromethyl, and aminomethyl;
R26 is hydrogen, (1-6C)alkyl, phenyl or benzyl;
R28 is cyano(3-6C)cycloalkenyl, or phenyl optionally substituted by 1 or 2 groups selected from halogen, nitro, amino, (1-4C)alkanoyl, (1-4C)alkanoylamino, halo(1-4C)alkyl, hydroxy, carboxy, (2-6C)alkoxycarbonyl, carbamoyl, mono- or di(1-4C)alkylcarbamoyl, cyano, mesyl, vinyl, and sulpho; or R28 is (2-6C)alkenyl, optionally substituted by halogen, cyano, carbamoyl, mono- or di(1-4C alkyl)carbamoyl, piperidinocarbonyl or morpholinocarbonyl, cyano(1-4C)alkyl, 2-ureidoethyl, 2-thioureidoethyl, 2-(thioacetyl-amino)ethyl, sulphamoyl, 2-amino-2-carboxyethyl, acetylaminomethyl, phthalimidomethyl, 4,5-dihydroimidazo1-2-ylmethyl, 3,4,5,6-tetrahydro-pyrimidin-2-ylmethyl, 2-(1,2,3,6-tetrahydro-2,6-dioxopurin-7-yl)ethyl, 2-hydroxyiminopropyl (syn or anti) or 2-[(1-4C)alkoxyimino]propyl (syn or anti),
or --R28 is of the formula --(CH.sub.2).sub.2 --NR29R30R31 in which R29, R30 and R31 are (1-4C)alkyl, or --R28 is of the formula --(CH.sub.2).sub.q --R32 in which q is 0-2 and --R32 is pyridine, pyridazine, pyrimidine, pyrazine, 1,2,5,6-dihydro-5,6-dioxo-1,2,4-triazine, 2-[(1-4C)alkyl]-1,2,5,6-dihydro-5,6-dioxo-1,2,4-triazine, 1-[(1-4C)alkyl]tetrazole, furan, thiophene, pyrrole, 1-[(1-4C)alkyl]pyrrole, oxazole, thiazole, imidazole, 1-[(1-4C)alkyl]imidazole, isoxazole, isothiazole, pyrazole, 1,2,3-thiadiazole, 1-[(1-4C)alkyl]pyrazole, benzfuran, benzthiophene, indole, oxindole, 1-[(1-4C)alkyl]indole, benzoxazole, benzthiazole, benzimidazole, 1-[(1-4C)alkyl]benzimidazole, 3,4-dihydro-4-oxo-2H-benzo[e]oxazine each of these ring systems being linked to (CH.sub.2).sub.q through carbon and each ring system being optionally substituted by halogen, amino, (1-6C)alkyl, (1-4C)-haloalkyl, (3-6C)cycloalkyl, (2-6C)alkenyl, carboxy, (2-6C)alkoxycarbonyl, (1-6C)alkoxy, cyano, (2-6C)cyanoalkenyl, carbamoyl, mono- or di (1-4C)alkylcarbamoyl, (1-4C)alkanoylamino, guanidino, hydroxy, nitro, amino; and for those ring systems which contain nitrogen, the N-oxides thereof where chemically possible;
or, when R4 is hydrogen then R5 is also a radical of the formula LI:
[Formula LI]
in which R28 is 2-guanidino-thiazol-4-ylmethyl, hydroxybenzoyl-methyl, 2-thenyl, 2-imidazolylmethyl or cinnamyl, optionally substituted by halogen, (1-6C)alkyl, hydroxy, (1-4C)alkoxy, carboxy, (2-6C)alkoxycarbonyl, nitro or carbamoyl,
and when R4 is other than hydrogen then R5 is also a radical of the formula LII in which ring J is pyridine, pyrimidine, oxazole, thiazole, isoxazole, isothiazole or imidazole to each of which is optionally fused, when possible, a benzene, cyclopentane or cyclohexane ring;
R33 is hydrogen, amino, (1-6C)alkyl, (3-6C)cycloalkyl, (3-6C)alkenyl, (2-8C)alkoxyalkyl, --(CH.sub.2).sub.t --COOR35, --(CH.sub.2).sub.t --CONH.sub.2, --(CH.sub.2).sub.t --NHCO--R36 or --(CH.sub.2).sub.t S(O).sub.s --R36 in which t is 1-6, R35 is hydrogen or (1-6C)alkyl, s is 0, 1 or 2, and R36 is (1-6C)alkyl or (1-6C)alkoxy,
or R33 is (3-8C)alkanoylmethyl, benzoylmethyl, (1-6C)primaryhydroxyalkyl, (1-6C)primaryaminoalkyl, (1-4C)alkylamino(1-6C)alkyl, di(1-4C)alkylamino(1-6C)alkyl, carbamoyl(1-4C)alkyl, mono- or di(1-4C)alkylcarbamoyl(1-4C)alkyl, (1-4C)alkoxy(1-4C)alkyl, (1-6C)alkoxy, (1-4C)alkoxy(2-4C) alkoxy(1-4C)alkyl, (1-6C)alkylamino, phenyl(1-6C)alkyl or phenyl(1-6C)alkoxy or of the formula (CH.sub.2).sub.2 N.dbd.CR37NR38R39 or (CH.sub.2).sub.n C(NR37)NR38R39 or a tautomer thereof in which R37, R38 and R39 are hydrogen or (1-4C)alkyl;
R34 is hydrogen or one or two substituents selected from halogen, amino, nitro, (1-6C)alkyl, carboxy, (2-6C)alkoxycarbcnyl, (1-6C)alkoxy, cyano, carbamoyl, (1-6C)haloalkyl, (1-6C)azidoalkyl, (1-6C)aminoalkyl, (2-4C)aminoalkylthio(1-4C)alkyl, (2-6C)alkanoylamino, (2-4C)alkanoylamino(1-4C)alkyl, (2-6C)alkanoyloxy(1-4C)alkyl, benzyl, benzyloxy and heteroarylthio,
wherein, when R33 contains phenyl, the phenyl is optionally substituted by halogen, nitro, (1-6C)alkyl, hydroxy, (1-4C)alkoxy, carboxy, (2-6C)alkoxycarbonyl, carbamoyl, sulphamoyl, sulpho, mono- or di(1-4C)alkylcarbamoyl, or mono- or di-(1-4C)alkylsulphamoyl, and wherein when R34 is heteroarylthio, the heteroaryl ring is a 5- or 6-membered ring containing 1,2 or 3 hetero atoms selected from oxygen, nitrogen and sulphur;
and the salts formed with acids and bases which afford pharmaceutically acceptable anions and cations respectively.
It is to be understood that in the above formula I and throughout this specification, the illustrated stereochemistry of the ceph-3-em nucleus, and its optional modifications at the 1-position, is the absolute configuration. It is also to be understood that, since R5 contains a quaternary nitrogen, the compounds of the formula I will normally exist in zwitter-ionic form, involving the quaternary nitrogen and the carboxy group. When the compound of the formula I contains further acidic or basic substituents, it is to be understood that the possibility of a double zwitter-ionic form of the compound will arise. Alternatively, exogenous anions or cations may be included, to form pharmaceutically-acceptable base-addition or acid-addition salts, as defined above.
A particular value for R2 is hydrogen, methyl, ethyl, isopropyl, t-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methylcyclopropyl, methylcyclobutyl, methylcyclopentyl, methylcyclohexyl, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, allyl, cyclopentenyl, cyclohexenyl, propargyl, methylcarbamoyl, ethylcarbamoyl, phenylcarbamoyl, benzylcarbamoyl, triphenylmethyl, 2-chloroethyl, 2-fluoroethyl, 2-bromoethyl, 2-hydroxyethyl, 3-hydroxypropyl, 2-methoxyethyl, 2-ethoxyethyl, 2-methylthioethyl, 2-methanesulphinylethyl, 2-methanesulphonylethyl, 2-aminoethyl, 3-aminopropyl, 2-methylaminoethyl, 2-dimethylaminoethyl, cyanomethyl, 2-cyanoethyl, azidomethyl, 2-azidoethyl, ureidomethyl, 3-amino-3-carboxypropyl, 2-(amidino)ethyl, 2-(N-aminoamidino)ethyl, tetrahydropyran-2-yl, thietan-3-yl or 2-oxo-tetrahydrofuran-3-yl,
or, when R2 is of the formula --(CH.sub.2).sub.n --R6 in which n is 1 to 4 and R6 is piperidino, pyrrolidino, morpholino, piperazino or N-methylpiperazino, a particular value of R2 is when each value of R6 is optionally substituted by methyl, phenyl or benzyl,
or, when R2 is of the formula --(CH.sub.2).sub.m --W--R7 in which m is 0 to 3, W is sulphur or a direct bond, particular values for R2 are when R7 is phenyl, pyridiniomethylene, 2-pyridinioethylene or R7 is pyridyl, imidazolyl, 1,3,4-thiadiazolyl, tetrazolyl, 1-methyltetrazolyl, thiazolyl, isothiazolyl or isoxazolyl in which the link with W is via a carbon or uncharged nitrogen, each value of R7 being optionally substituted, where possible, by one or two groups selected from methyl, amino, hydroxy, carboxy, carbamoyl, nitro, methoxycarbonyl, ethoxycarbonyl, cyano or sulpho,
or, when R2 is of the formula --(CH.sub.2).sub.n --CO--R8 in which n is 1 to 4, a particular value for R2 is when R8 is methyl, ethyl, phenyl or benzyl,
or, when R2 is of the formula --COR9 or --(CH.sub.2).sub.n --OCO--R9 in which n is 1-4, a particular value for R2 is when R9 is hydrogen, methyl, chloromethyl, bromomethyl, 2-chloroethyl, 2-bromoethyl, phenyl or benzyl,
or, when R2 is of the formula --G--CH.sub.2 --R10, a particular value for R2 is when G is carbonyl or a direct bond and RIO is phthalimido,
or, when R2 is of the formula --NR11R12R13, a particular value for R2 is when R11, R12 and R13 are methyl or ethyl, or R11 is methyl or ethyl and R12 and R13 are joined to form a cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl ring, or R11, R12 and R13 are joined to form a 1-azonia-4-azabicyclo[2,2,2]octane or 1-azonia-3,5,7-triazatricyclo[3,3,1,1.sup.3,7 ]decane,
or, when R2 is of the formula II:
[Formula II]
in which p is 1 or 2, a particular value for R2 is when and R14 and R15 are hydrogen or methyl,
or when R2 is of the formula --P(O)R16R17, a particular value for R2 is when R16 is hydroxy, methoxy, ethoxy, dimethylamino, diethylamino, phenoxy, phenylamino, or one of the particular values given above for R6, and R17 is methyl, ethyl, methoxy, ethoxy, dimethylamino, diethylamino, phenoxy, phenylamino, piperidino, pyrrolidino, morpholino, piperazino or N-methylpiperazino,
or, when R2 is of the formula --CH.sub.2 P(O)R18R19, a particular value for R2 is when R18 and R19 are hydroxy, methoxy or ethoxy,
or, when R2 is of the formula --CH(SR20)COOR21, a particular value for R2 is when R20 is methyl or ethyl and R21 is hydrogen, methyl, ethyl or isopropyl,
or, when R2 is of the formula III:
[Formula III]
in which m is 0-3, a particular value for R2, when m=0, is when R22 is hydrogen, methyl or methylthio, R23 is hydrogen, methyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyano, carboxy, carboxymethyl, 2-carboxyethyl or methanesulphonylamino, or phenyl optionally substituted by amino or hydroxy, or R23 and R24 are joined to form, together with the carbon to which they are attached, a cyclopropane, cyclobutane, cyclopentane, cyclohexane or cycloheptane ring and R25 is hydroxy, amino, methoxy, ethoxy, methylamino, ethylamino, phenylamino or one of the particular values for R6 given above or of the formula NHOR25 in which R25 is hydrogen, methyl, ethyl, phenyl or benzyl,
provided that when R2 contains phenyl, and unless otherwise stated above, the phenyl is optionally substituted by 1 or 2 groups selected from fluorine, chlorine, bromine, hydroxy, amino, carboxy, nitro, carbamoyl, cyano and aminomethyl.
A particular value for R3 is hydrogen or methoxy.
A particular value for R4 is hydrogen, methyl, ethyl, n-propyl, isopropyl, 2-fluoroethyl, 2-chloroethyl, 2-hydroxymethyl, 2-methoxyethyl, carboxymethyl, (R) and (S)-1-carboxyethyl, 2-aminoethyl, 2-cyanoethyl, 2-formamidoethyl, allyl, furfuryl, benzyl or 4-pyridylmethyl.
A particular value for an optional substituent on one of the ring systems of the formula IV to LI inclusive is one, two or three substituents selected from fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, carboxy, methoxycarbonyl, ethoxycarbonyl, methoxycarbonylmethyl, ethoxycarbonylmethyl, methoxy, ethoxy, methylthio, ethylthio, cyano, cyanomethyl, 2-cyanoethyl, amino, methylamino, ethylamino, isopropylamino, dimethylamino, benzylamino, (optionally substituted in the benzene ring by nitro), allylamino, 2-aminoethyl-amino, 2-methoxyethylamino, 2-hydroxyethylamino, hydroxy, mercapto, carbamoyl, methylcarbamoyl, ethylcarbamoyl, dimethylcarbamoyl, phenylthio and heteroarylthio in which the heteroaryl ring is a furan, thiophene, imidazole, thiazole, pyrazole, thiadiazole, pyridine, pyrimidine, pyrazine or pyridazine.
A particular value for R27 is methyl, ethyl, n-propyl, isopropyl, allyl, methoxymethyl, 2-methoxyethyl, carboxymethyl, 2-carboxyethyl, methoxycarbonylmethyl, 2-methoxycarbonylethyl, carbamoylmethyl, 2-carbamoylethyl, carboxyaminocarbonylmethyl, 2-(carboxyaminocarbonyl)ethyl, methoxycarbonylaminocarbonylmethyl, 2-(methoxycarbonylaminocarbonyl)ethyl, acetylmethyl, propionylmethyl, benzoylmethyl, hydroxymethyl, 2-hydroxyethyl, methylamino, ethylamino, benzyl or 2-phenethyl, or phenyl optionally substituted by 1 or 2 groups selected from fluorine, chlorine, bromine, hydroxy, amino, carboxy, nitro, carbamoyl, cyano, trifluoromethyl and aminomethyl.
A particular value for R26 is hydrogen, methyl, ethyl, n-propyl, isopropyl, phenyl or benzyl.
A particular value for R28 is 3-cyanocyclopent-2-enyl or phenyl optionally substituted by 1 or 2 groups selected from fluorine, chlorine, bromine, nitro, amino, acetyl, acetamido, trifluoromethyl, hydroxy, carboxy, methoxycarbonyl, ethoxycarbonyl, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, diethylcarbamoyl, cyano, mesyl and sulpho,
or vinyl, 2,4-pentadienyl, 3-chloroallyl (cis and trans), 3-cyanoallyl, cyanomethyl, 3-cyanopropyl, 2-ureidoethyl, 2-thioureidoethyl, 2-(thioacetylamino)ethyl, sulphamoyl, 2-amino-2-carboxyethyl, acetylaminomethyl, phthalimidomethyl, 4,5-dihydroimidazol-2-ylmethyl, 3,4,5,6-tetrahydropyrimidin-2-ylmethyl, 2-(1,2,3,6-tetrahydro-2,6-dioxopurin-7-yl)ethyl, 2-hydroxyiminopropyl (syn or anti), 2-(mathoxyimino)propyl (syn or anti) or 2-(ethoxyimino)propyl (syn or anti),
or of the formula --(CH.sub.2).sub.2 --NR29R30R31 in which R29, R30 and R31 are methyl or ethyl,
or of the formula --(CH.sub.2).sub.q --R32 in which q is 0-2 and R32 is pyridine, pyridazine, pyrimidine, pyrazine, 1,2,5,6-dihydro-5,6-dioxo-1,2,4-triazine, 2-(methyl or ethyl)-1,2,5,6-dihydro-5,6-dioxo-1,2,4-triazine, 1-(methyl or ethyl)tetrazole, furan, thiophene, pyrrole, 1-(methyl or ethyl)pyrrole, oxazole, thiazole, imidazole, 1-(methyl or ethyl)imidazole, isoxazole, isothiazole, pyrazole, 1-(methyl or ethyl) pyrazole, 1,2,3-thiadiazole, benzfuran, benzthiophene, indole, 1-(methyl or ethyl)indole, benzoxazole, benzthiazole, benzimidazole, 3,4-dihydro-4-oxo-2H-benzo[eloxazine, 1-(methyl or ethyl)benzimidazole, each of these ring systems being linked to (CH.sub.2).sub.q through carbon and each ring system being optionally substituted by fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, trifluoromethyl, cyclopropylmethyl, formamido, carboxy, methoxycarbonyl, ethoxycarbonyl, methoxy, ethoxy, cyano, 3-cyanoallyl, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, hydroxy, guanidino, nitro, amino and for those ring systems which contain nitrogen, the N-oxides thereof where chemically possible;
or, when R4 is hydrogen --R28 is 2-guanidinothiazol-4-ylmethyl, 3-hydroxybenzoylmethyl, 2-thenyl, 2-imidazolylmethyl or cinnamyl, optionally substituted by fluorine, chlorine, bromine, methyl, ethyl, n-propyl, isopropyl, hydroxy, methoxy, ethoxy, carboxy, methoxycarbonyl, ethoxycarbonyl or carbamoyl;
and when R4 is other than hydrogen, a further particular value for R5 is of the formula LI:
[Formula LII]
in which ring J is pyridine, pyrimidine, oxazole, thiazole, isoxazole, isothiazole or imidazole to each of which is optionally fused, when possible, a benzene, cyclopentane or cyclohexane ring.
A particular value for R33 is hydrogen, amino, methyl, ethyl, n-propyl, isopropyl, t-butyl, cyclopropyl, cyclobutyl, allyl, 3-chloroallyl (cis or trans) methoxymethyl, --(CH.sub.2).sub.t --COOR35, (CH.sub.2).sub.t --CONH.sub.2, (CH.sub.2).sub.t --NH--CO--R36 or --(CH.sub.2).sub.t S(O).sub.s --R36 in which t is 1-6, R35 is hydrogen, methyl or ethyl, s is 0, 1 or 2 and R36 is methyl, ethyl, methoxy or ethoxy,
or acetylmethyl,propionylmethyl, benzoylmethyl, hydroxymethyl, 2-hydroxyethyl, aminomethyl, 2-aminoethyl, 2-methylaminoethyl, 2-dimethylaminoethyl, carbamoylmethyl, methylcarbamoylmethyl, dimethylcarbamoylmethyl, 2-methoxymethyl, methoxy, ethoxy, 2-methoxyethoxymethyl, methylamino, ethylamino, benzyl, 2-phenethyl, 2-(3,4-dihydro-4-oxo-H-benzo[e]- oxazin-2-yl)ethyl, benzyloxy or 2-phenylethoxy or of the formula (CH.sub.2).sub.2 --N.dbd.CR37NR38R39 or (CH.sub.2).sub.2 C(N37)NR38R39 in which R37, R33 and R39 are hydrogen or methyl, wherein when R33 contains phenyl the phenyl is optionally subtituted by fluorine, chlorine, bromine, methyl, ethyl, hydroxy, methoxy, ethoxy, carboxy, methoxycarbonyl, ethoxycarbonyl, sulpho, carbamoyl, methylcarbamoyl or dimethylcarbamoyl.
A particular value for R34 is hydrogen or one or two substituents selected from fluorine, chlorine, bromine, amino, nitro, methyl, ethyl, carboxy, methoxycarbonyl, ethoxycarbonyl, methoxy, ethoxy, cyano, carbamoyl, chloromethyl, 2-chloroethyl, azidomethyl, aminomethyl, 2-aminoethyl, 2-aminoethylthiomethyl, acetylamino, propionylamino, acetylaminomethyl, acetoxymethyl, benzyl, benzyloxy, 2-thenylamino or heteroarylthio and in which when R34 is heteroarylthio the heteroaryl ring is furan, thiophene, pyrrole, imidazole, thiazole, isothiazole, oxazole, isoxazole, pyrazole, thiadiazole, pyridine,
A particular acid which affords a pharmaceutically-acceptable anion is, for example, hydrochloric, hydrobromic, phosphoric, sulphuric, citric or maleic acid.
A particular base which affords a pharmaceutically acceptable cation is, for example, a base containing an alkali metal, (e.g. sodium or potassium) or an alkaline earth metal (e.g. calcium or magnesium), or a primary, secondary or tertiary organic amine (e.g. triethylamine, morpholine, N-methylpiperidine, N-ethylpiperidine, procaine, dibenzylamine or N,N'-dibenzylethylenediamine), or other amine which has been used to form salts with cephalosporins.
The following are preferred features of the cephalosporin derivative of the invention. When any one of these features is taken, either singly or in combination, with the other general or particular features of the cephalosporin derivative of the invention listed above, there are obtained preferred sub-groups of compounds.
1. X is sulphur.
2. R1 is 2-aminothiazol-4-yl or 5-amino-1,2,4-thiadiazol-3-yl.
3. R50 is chloromethylene.
4. R50 is .dbd.N.OR2 in which R2 is (1-6C)alkyl, (3-6)alkenyl optionally substituted by carboxy, (3-6C)alkynyl, (3-8C)cycloalkyl, (3-6C)cycloalkyl(1-3C)alkyl, (1-4C)haloalkyl, (1-5C)cyanoalkyl, (2-6C)hydroxyalkyl, (1-4C)alkoxy(2-4C)alkyl, (2-6C)aminoalkyl or benzyl.
5. R2 is methyl, ethyl, i-propyl, allyl, propargyl, cyclopentyl, cyclopropylmethyl, 2-chloroethyl, 2-fluoroethyl, 2-bromoethyl, cyanomethyl, 2-cyanoethyl, 2-hydroxyethyl, 2-ethoxyethyl or benzyl.
6. R2 is of the formula III.
7. In formula III, m is 0.
8. In formula III, R24 is hydroxy or (1-4C)alkoxy.
9. In formula III, R22 and R23 are both hydrogen or (1-3C)alkyl, or R22 and R23 are joined to form, together with the carbon to which they are attached, a (3-7C) carbocyclic ring.
10. In formula III, R22 and R23 are both hydrogen or methyl, or R22 and R23 are joined to form, together with the carbon to which they are attached, a cyclobutane or cyclopentane ring.
11. R5 is of the formula XVI, XXVIII, XLII, XLIII, XLVI, XLVII or LI, each of these ring systems being optionally substituted, where possible, on a carbon atom or atoms, by one or two substituents selected from halogen, (1-6C)alkyl, carboxy, (2-6C)alkoxycarbonyl(1-4C)alkyl, (1-6C)alkoxy, (1-6C)alkylthio, amino, (1-6C)alkylamino, (2-8C)dialkylamino, benzylamino, optionally substituted in the benzene ring thereof by nitro, allylamino, (1-6C)aminoalkylamino, (1-6C)- alkoxy(1-6C)alkylamino, (1-6C)hydroxyalkylamino and hydroxy.
12. In formulae XVI, XXVIII, XLII, XLIII, XLVI and XLVII, the ring substituents are selected from chlorine, fluorine, methyl, ethyl, n-propyl, isopropyl, carboxy, methoxycarbonylmethyl, ethoxycarbonylmethyl, methoxy, ethoxy, methylthio, amino, isopropylamino, dimethylamino, p-nitrobenzylamino, allyamino, 2-aminoethylamino, 2-methoxyethylamino, 2-hydroxyethylamino and hydroxy.
13. In formulae XVI, XXVIII and XLVII, R27 is (1-6C)alkyl, allyl or phenyl optionally substituted by nitro or trifluoromethyl.
14. In formulae XLII and LI, R28 is phenyl optionally substituted by 1 or 2 groups selected from halogen, nitro, amino, (1-4C)alkanoyl, (1-4C)alkanoylamino, (1-4C)haloalkyl, hydroxy, (2-6C)alkoxycarbonyl, carbamoyl, mono- or di(1-4C)alkylcarbamoyl, cyano, mesyl and vinyl.
15. In formulae XLII and LI, the substituents in R28, when R28 is phenyl, are selected from chlorine, fluorine, nitro, amino, acetyl, acetamido, trifloromethyl, hydroxy, methoxycarbonyl, ethoxycarbonyl, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, cyano, mesyl and vinyl.
16. In formulae XLII and LI, R28 is (2-6C)alkenyl, optionally substituted by halogen or cyano, cyano(1-4C)alkyl, phthalimidomethyl or 2-(1,2,3,6-tetrahydro-2,6-dioxopurin-7-yl)ethyl.
17. In formulae XLII and LI, R28 is vinyl, 2,4-pentadienyl, 3-cyanopropyl, 3-chloroallyl, 3-cyanoallyl, phthalimidomethyl or 2-(1,2,3,6-tetrahydro-2,6-dioxopurin-7-yl)ethyl.
18. In formulae XLII and LI, R28 is of the formula (CH.sub.2).sub.q --R32, wherein q is 0 or 1 and R32 is pyridine, pyridine-N-oxide, pyridazine, pyrazine, pyrazine-N-oxide, pyrimidine, furan, thiophene, thiazole, imidazole, 1,2,3-thiadiazole, oxazole, oxindole, benzimidazole or 3,4-dihydro-4-oxo-2H-benzo[e]-oxazine, optionally substituted by halogen, amino, (1-6C)alkyl, (1-4C)haloalkyl, (3-6C)cycloalkyl, (2-6C)alkoxycarbonyl, cyano, (2-6C)cyanoalkenyl, carbamoyl, mono- or di(1-4C)alkylcarbamoyl, (1-4C)alkanoylamino, guanidino, hydroxy, nitro or amino.
19. The substituent in R32 is chlorine, fluorine, amino, methyl, ethyl, n-propyl, isopropyl, trifluoromethyl, cyclopropyl, methoxycarbonyl, ethoxycarbonyl, cyano, 3-cyanoallyl, carbamoyl, methylcarbamoyl, dimethylcarbamoyl, formamido, guanidino, hydroxy, nitro or amino.
20. In formula LII, R33 is (1-6C)alkyl and R34 is hydrogen.
21. R33 is methyl or ethyl.
22. R4 is hydrogen, (1-4C)alkyl, halo(1-4C)alkyl, hydroxy(1-4C)alkyl, (1-4C)alkoxy(1-4C)alkyl, carboxy(1-4C)alkyl, amino(1-4C)alkyl, cyano(1-4C)alkyl, (1-4C)alkanoylamino(1-4C)alkyl, allyl, furfuryl, benzyl or pyridyl (1-4C)alkyl.
23. R4 is hydrogen, methyl, ethyl, 2-fluoroethyl, 2-chloroethyl, 2-hydroxyethyl, 2-methoxyethyl, carboxymethyl, 2-aminoethyl, 2-cyanoethyl, 2-formamidoethyl, allyl, furfuryl, benzyl or 4-pyridylmethyl.
24. R5 is of the formula XVI, XXVIII, XLII, XLIII, XLVI, LI or LII.
25. R5 is of the formula XVI wherein the ring is substituted by (1-6C)alkyl and/or amino.
26. R5 is of the formula XXVIII wherein Y is sulphur, Z is CH and R27 is (1-6C)alkyl, particularly methyl.
27. R5 is of the formula XLII wherein R29 is (2-6C)alkenyl, optionally substituted by cyano or halogen, particularly chlorine, or R28 is (CH.sub.2).sub.q R32 in which q is 1 and R32 is 1,2,3-thiadiazole, thiophen or thiazole optionally substituted by guanidino or cyano.
28. R5 is of the formula XLIII wherein Y is sulphur.
29. R5 is of the formula XLVI wherein the optional bond is a double bond.
30. R5 of the formula LI wherein R28 is (CH.sub.2).sub.q R32 in which q is 1 and R32 is thiophen.
31. R5 is of the formula LII wherein ring J is pyridine, R33 is amino, (1-6C)alkyl, (1-6C)primary-aminoalkyl, or benzyl optionally substituted by nitro, and R34 is hydrogen or (1-6c)alkyl.
Particular compounds of the invention are described in the Examples, and of these, the group consisting compounds of Examples 37, 55, 60, 64, 66, 71, 81, 87, 88, 95, 96, 104, 110, 111, 149, 150, 151, 153, 177, 178, 184, 185, 188, 195, 202, 209, 213, 215, 218, 219, 225, 226, 232, 244, 249, 251, 255, and 261 is preferred. A more preferred group consists of the compounds of Examples 37, 55, 60, 64, 66, 71, 88, 96, 104, 110, 149, 150, 153, 178, 184, 213, 219, 225, 244, 249, 255, and 261. Within this group, the compounds of Examples 60, 66, 104, 110, 184, 188, 219, 225, 255 and 261 are particularly preferred, and of these the compounds of Examples 110, 184, 219 and 225 are especially preferred.
The cephalosporin derivatives of the formula I may be manufactured by methods known in themselves for the manufacture of chemically analogous compounds. The for following processes, R1, R2, R3, R4, R5, R50 and X having the meanings stated above, unless indicated otherwise, and therefore provided as further features of the invention.
The process of the invention is characterised by
(a) reaction of a compound of the formula LIII
[Formula LIII]
with a compound of the formula R5-R40 in which R40 is a displaceable radical [e.g. fluorine, chlorine, bromine, (1-6C)alkoxy, (1-6C)alkylthio, (1-6C)alkanesulphinyl or (1-6C)alkanesulphonyl].
(b) reaction of a compound of the formula LIV:
[Formula LIV]
with an acid of the formula LV:
[Formula LV]
or an activated derivative thereof.
(c) deprotection, to form carboxy, of the corresponding compound which carries a protecting group in place of the acidic hydrogen atom of the carboxy
(d) deprotection, to form a primary or secondary amino, of the corresponding compound which carries a protecting group in place of the amino hydrogen.
(e) for those compounds in which X is sulphinyl, oxidation of the corresponding compound in which X is sulphur.
(f) reaction of a compound of the formula LVI
[Formula LVI]
with a compound of the formula R2--O--NH.sub.2.
(g) for those compounds in which R2 is other than hydrogen, reaction of a compound of the formula I in which R2 is hydrogen with a compound of the formula R41-R40 in which R40 is a displaceable radical and R41 is one of the values given above for R2, other than hydrogen.
(h) for those compounds which contain an aminophenyl group, the reduction of the corresponding nitrophenyl compound.
(i) the reaction of a compound of the formula LVIII, wherein R40 is a displaceable radical, with a compound of the formula R4R5NH.
When the process of the invention manufacture the compound of the formula I in the form of the zwitterion, and a salt is required, the compound of the formula I in the zwitterionic form is reacted with an acid which affords a pharmaceutically-acceptable anion, or with a base which affords a pharmaceutically-acceptable cation.
The starting material of the formula LIII may be prepared by acylation of the appropriate 7-amino-3-azidomethylcephalosporin derivative with an acid of the formula LV, or an activated derivative thereof, followed by reduction of the 3-azidomethyl group to the 3-aminomethyl group. During this process it may be necessary to protect amino and carboxy groups. There is thus obtained the compound of the formula LIII in which R4 is hydrogen. This procedure is illustrated in Examples 1-4, 5-6, 7-14 and 15-16. When R4 is other than hydrogen, the compound in which R4 is hydrogen is then alkylated or benzylated.
The starting material of the formula LIV may be prepared by reaction of the compound of the formula LVII:
[Formula LVII]
with a compound of the formula R5-R40 in which R40 is a displaceable radical. During this reaction it may be necessary to protect the 7-amino and/or 3-carboxy group.
The starting material of the formula LIV in particularly valuable and is regarded as a further feature of the invention.
The starting materials of the formulae LV, LVI, LVII and LVIII are prepared by conventional methods known in the chemical literature for analogous compounds.
As noted above the cephalosporin derivatives of the invention have antibacterial properties. Thus they are useful antibacterial agents, many of them having a broad spectrum of activity in vitro against standard laboratory microorganisms, both Gram-negative and Gram- positive, which are used to screen for activity against pathogenic bacteria. The antibacterial spectrum and potency of a particular compound may be determined in a standard test system.
The antibacterial properties of the compounds of the invention may also be demonstrated in conventional mouse protection tests, and the preferred compounds of this invention have MIC.sub.50 values of less than 2 .mu.g./ml. against both Pseudomonas aeruginosa and Staphyllococcus aureus.
Cephalosporin derivatives have generally been found to be relatively non-toxic to warm-blooded animals, and this generalisation holds true for the compounds of the present invention. A number of compounds were administered to mice at doses in excess of those required to afford protection against bacterial infections, and no overt toxic symptoms or side effects attributable to the administered compounds were noted.
According to a further feature of the invention there is provided a pharmaceutical composition which comprises a cephalosporin derivative of the invention in association with a pharmaceutically-acceptable diluent or carrier.
The pharmaceutical composition of the invention may, for example, be in a form suitable for oral, rectal or parenteral administration, for which purposes it may be formulated by means known to the art into the form of, for example, tablets, capsules, aqueous or oily solutions or suspensions, emulsions, dispersible powders, suppositories and sterile injectable aqueous or oily solutions or suspensions.
In addition to the cephalosporin derivative of the formula I the pharmaceutical composition of the invention may also contain, or be co-administered with, one or more known drugs selected from other clinically useful antibacterial agents (for example other beta-lactams or aminoglycosides), inhibitors of beta-lactamase (for example clavulanic acid), renal tubular blocking agents (e.g. probenicid) and inhibitors of metabolising enzymes (for example inhibitors of peptidases, for example Z-2-acylamino-3-substituted propenoates).
A preferred pharmaceutical composition of the invention is one suitable for intravenous, subcutaneous or intramuscular injection, for example a sterile injectable containing between 1 and 10% w/w of the cephalosporin derivative, or one suitable for oral administration in unit dosage form, for example a tablet or capsule which contains between 100 mg. and 1 g. of the cephalosporin derivative.
The pharmaceutical composition of the invention will normally be administered to man in order to combat infections caused by bacteria, in the same general manner as that employed for cephalothin, cefoxitin, cephradine and other known clinically used cephalosporin derivatives, due allowance being made in terms of dose levels for the potency of the cephalosporin derivative of the present invention relative to the known clinically used cephalosporins. Thus each patient will receive a daily intravenous, subcutaneous or intramuscular dose of 0.5 to 50 g., and preferably 0.5 to 10 g., of the cephalosporin derivative, the composition being administered 1 to 4 times per day. The intravenous, subcutaneous and intramuscular dose will be given by means of a bolus injection. Alternatively the intravenous dose may be given by continuous infusion over a period of time. Alternatively each patient will receive a daily oral dose which is approximately equivalent to the daily parenteral dose. Thus a preferred daily oral dose is 0.5 to 10 g. of the cephalosporin derivative, the composition being administered 1 to 4 times per day.
The invention is illustrated, but not limited, by the following Examples. The n.m.r. spectra are quoted in delta relative to tetramethylsilane (delta=0) as internal standard, (s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, br=broad). The n.m.r. are measured at a field strength of 90 or 400 MHz. The n.m.r. solvents are as follows:
Solvent A:-d.sub.6 DMSO+CD.sub.3 COOD
Solvent B:-d.sub.6 DMSO+CD.sub.3 COOD+TFA
Solvent C:-CDCl.sub.3 +CD.sub.3 COOD
The temperatures are in degrees Centigrade. The following contractions are used:
______________________________________TFA = trifluoroacetic acidTHF = tetrahydrofuranHOAc = acetic acidEtOAc = ethyl acetateMeOH = methanolDMF = dimethylformamideDMSO = dimethylsulphoxideether = diethyl etherHPLC = high pressure liquid chromatographyEt.sub.3 N = triethylamineLawesson's reagent = 2,4-dithioxo-P.sup.5,P.sup.5 - 1,3,2,4-dithia- phosphatane.MCBPA = m-chloroperbenzoic acid.DCCI = N,N'dicyclohexyl- carbodi-imide.CAZAMCA = ##STR3##BOC = tert-butyoxycarbonyl.______________________________________

EXAMPLES 1-4
To a solution of 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-[(Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid (0.4 mmole) in DMF (11 ml.) and water (3 ml.) at 0.degree. was added sodium bicarbonate (4.8 mmole) dissolved in the minimum volume of water followed by the appropriate 4-chloroheterocycle (0.4 mmole). After 1 hour the mixture was treated with HOAC (4.8 mmole) and evaporated to dryness under reduced pressure. The residue was dissolved in water and purified by HPLC on an octadecylsilane column using MeOH/water/HOAC 40:60:1 v/v/v eluant. Using this general method the following compounds were thus obtained
______________________________________ ##STR4##Example R Yield % Footnotes______________________________________ ##STR5## 39 1, 22 ##STR6## 31 3, 43 ##STR7## 33 5, 64 ##STR8## 31 5, 7______________________________________Footnotes1. The starting material was prepared byreaction of 1-phenyl-4-pyridone withtoluene- - p-sulphonylchloride in refluxing toluene togive 4-chloro-1-phenyl toluene- -p-sulphonate.2. n.m.r. in solvent A: 1.5(s, 6H); 3.35(d, 1H); 3.68(d, 1H); 4.3(d, 1H); 4.6(d, 1H); 5.12(d, 1H); 5.75(d, 1H); 6.68(s, 1H); 7.1(d, 1H); 7.6(m,6H); 8.4(d, 1H); 8.5(d, 1H).3. The starting material was prepared asfollows. A mixture of 2,6-dicarboxy-4-pyrone (1.84 g.)and 4-fluoroaniline (1.9 ml.) was heated at 180.degree. underargon until evolution of gas ceased. The cooledmixture was purified by chromatography on silica gel,eluting with CH.sub.2 Cl.sub.2 /MeOH 100-90:0-10 v/v to give, aftersublimation (150.degree./0.01 mmHg) 1-(4-fluorophenyl)-4-pyridone(1.24 g.). n.m.r. in CDCl.sub.3 : 6.45(d, 2H); 7.3(m, 4H); 7.54(d, 2H). Reaction of this compound withtoluene- -p-sulphonylchloride in refluxing toluene gave4-chloro-1-(4-fluorophenyl)pyridiniumtoluene- -p-sulphonate.4. n.m.r. in solvent A: 1.4(s, 6H); 3.35(d, 1H); 3.65(d, 1H); 4.25(d, 1H); 4.46(d, 1H); 5.1(d, 1H); 5.8(d, 1H); 6.7(s, 1H); 6.9-7.8(complex, 6H);8.3(d, 1H); 8.5(d, 1H).5. The starting material was prepared by themethod described in Footnote 3, using the appropriatesubstituted aniline.6. n.m.r. in solvent A: 1.4(s, 6H); 3.36(d, 1H); 3.64(d, 1H); 4.27(d, 1H); 4.48(d, 1H); 5.1(d, 1H); 5.8(d, 1H); 6.7(s, 1H); 6.9-7.7(m, 6H); 8.35(d, 1H); 8.5(d, 1H).7. n.m.r. in solvent A: 1.45(s, 6H); 3.4(d, 1H), 3.68(d, 1H); 4.3(d, 1H); 4.5(d, 1H); 5.12(d, 1H); 5.85(d, 1H); 6.75(s, 1H); 6.9-7.9(m, 5H); 8.2(d, 1H); 8.4(d, 1H).
The cephalosporin starting material may be obtained as follows:
To a stirred mixture of DMF (5.80 ml.) in anhydrous methylene chloride (415 ml.) at -10.degree. was added dropwise oxalyl chloride (6.15 ml.). Stirring was continued at -10.degree. for 30 minutes to give a gelatinous white precipitate of (chloromethylene)dimethylammonium chloride. To this stirred suspension was added powdered 2-((Z)-1-t-butoxycarbonyl-1-methylethoxyimino)-2-(2-tritylaminothiazol-4-yl)acetic acid (40.0 g.) followed by N-methylmorpholine (8.8 ml.). Stirring was continued for 30 minutes between -5.degree. and -15.degree..
In another flask a suspension of 7-amino-3-azidomethylceph-3-em-4-carboxylic acid (17.85 g.) in anhydrous methylene chloride (150 ml.)was stirred for 1 hour with N,O-bis(trimethylsilyl)acetamide (34.5 ml.) to give a clear orange solution. This was transferred by syringe to the above acid chloride solution which was stirred at -10.degree. during the addition. The reaction mixture was then allowed to warm to room temperature and stirred for a further 90 minutes. The mixture was then poured into water (500 ml.) and extracted with EtoAc (3.times.500 ml.). The combined EtOAc extracts were washed with water, dried (Na.sub.2 SO.sub.4) and the solvent was evaporated under reduced pressure to yield a buff foam. The crude product was dissolved in methylene chloride and applied to a column of Kieselgel 60 (125 g.). Elution with methylene chloride/MeOH/HOAc 96:2:2 v/v/v gave 3-azidomethyl-7-[2-(2-tritylaminothiazol-4-yl)-2-((Z)-1-t-butoxycarbonyl-1-methylethoxyimino)acetamido]ceph- 3-em-4-carboxylic acid (46.4 g.) as a white foam n.m.r. in solvent A: 1.30 (s, 9H); 1.35 (s,6H), 3.37 (d, 1H), 3.65 (d, 1H), 3.90 (d,1H), 4.35 (d,1H), 5.1 (d,1H); 5.7 (d,1H); 6.66 (s,1H), 7.25 (s,15H).
An aqueous slurry of Raney nickel (10.2 g.) was added in one portion to a stirred solution of the azide (20.0 g.) in a mixture of MEOH (60 ml.) and TFA (60 ml.) at room temperature. A vigorous effervescence was observed. Stirring was continued for 1 hour and the Raney nickel was removed by filtration through diatomaceous earth. The filter pad was washed well with MEOH and the washings were combined with the filtrate. The solvents were evaporated under reduced pressure to give a pale green solid residue which was then stirred for 2 hours with a mixture of TFA (60 ml.) and water (15 ml.). This mixture was evaporated to dryness and the residue was stirred vigorously with water (400 ml.) for 30 minutes. The resulting solution was filtered through diatomaceous earth to remove the undissolved triphenylmethanol and the filtrate was applied to a column of Diaion HP 20 resin (1 l.). The column was eluted with water (500 ml.) to remove inorganic material and then with aqueous MEOH 1:1 v/v. The fractions which were shown by HPLC to contain the product were evaporated under reduced pressure to yield 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid (4.2 g.) as a pale yellow foam having the following n.m.r. in solvent A: 1.4 (s,6H); 3.1-3.8 (complex, 4H); 4.95 (d,1H); 5.7(d,1H); 6.72 (s,1H).
EXAMPLES 5-6
The general process described in Examples 1-4 was repeated, and the following compounds were obtained.
__________________________________________________________________________ ##STR9##Example No. R Yield % Footnotes__________________________________________________________________________5 C.sub.2 H.sub.5 64 1, 2, 3 ##STR10## 55 4, 5__________________________________________________________________________Footnotes1. The starting material may be prepared as follows. Reaction of (.sub.-- Z)-2-ethoxyimino-2-(2-trityl-aminothiazol-4-yl)acetic acid and 7-amino-3-azidomethyl-ceph-3-em-4-carboxylic acid (previously treated with N,O-bistrimethylsilylacetamide in the presence of chloromethyleneammoniumchloride and N-methylmorpholine) in CH.sub.2 Cl.sub.2 /DMF gave afterpurification by chromatography on silica gel 3-azidomethyl-7-[( .sub.--Z)-2-ethoxyimino-2-(2-tritylaminothiazol-4-yl)acetamido]ceph-3-em-4-carboxylicacid; n.m.r. in CDCl.sub. 3 : 1.29(t, 3H); 3.3(d, 1H); 3.54(d, 1H);3.87(d,1H), 4.3(q, 2H); 4.38(d, 1H); 4.97(d, 1H); 5.73(q, 1H); 6.72(s, 1H);7.26(m, 15H).This 3-azidomethyl compound in TFA acid was reduced with Raney nickel togive, after purification by chromatography on a Diaion CHP20Pcolumn, 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-( .sub.-- Z)-2-ethoxyimino)-acetamido]ceph-3-em-4-carboxylic acid; n.m.r. in solvent A:1.2(t, 3H); 3.26(d, 1H); 3.48(br s, 2H); 3.68(d, 1H); 4.08(q, 2H);4.97(d, 1H),5.66(d, 1H); 6.69(s, 1H).2. The quaternised starting material was 3-chloro-1-methylpyridaziniumiodide.3. n.m.r. in solvent A: 1.2(t, 3H); 3.31(d, 1H); 3.61(d, 1H); 4.12(q,2H);4.23(s, 3H); 4.48(d, 1H); 4.99(d, 1H); 5.65(d, 1H); 6.69(d, 1H); 7.6(m, 1H); 7.9(dd, 1H); 8.88(d, 1H).4. n.m.r. in solvent A: 1.9(m, 2H); 2.45(m, 4H); 3.44(d, 1H); 3.66(d,1H);4.1(d, 1H); 4.28(s, 3H); 4.57(d, 1H); 5.1(d, 1H); 5.82(d, 1H); 6.83(s, 1H); 7.63(br, 1H); 7.92(br, 1H); 8.9(d, 1H).5. The starting material was prepared from 2-[( .sub.-- Z)-1-(t-butoxycarbonyl)cyclobut-1-yloxyimino]-2-(2-tritylaminothiazol-4-yl)acetic acid by the general method described in Footnote 1. to give3-azido-methyl-7-(( .sub.-- Z)-2-[ 1- .sub.- t-butoxycarbonyl)cyclobut-1-yloxyimino]-2-[2-tritylaminothiazol-4-yl]acetamido)ceph-3-em-4-carboxylic acid, having the following n.m.r. in solvent A: 1.38(s, 9H);1.78(m, 1H); 1.89(m, 1H); 2.35(m, 4H); 3.45(d, 1H); 3.63(d, 1H); 3.9(d,1H); 4.38(d, 1H); 5.13(d, 1H); 5.76(d, 1H); 6.68(s, 1H); 7.18-7.37(m,15H).
The above 3-azidomethylcephalosporin derivative was reduced by the general method described in Footnote 1 to give 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-(Z)-2-(1-carboxycyclobut-1-yloxyimino)acetamido]ceph-3-em-4-carboxylic acid, which was partially purified by chromatography on Diaion CHP20 resin before use without further characterisation.
EXAMPLES 7-14
The general process described in Examples 1-4 was repeated using the appropriate heterocyclic starting material. The reactions were carried out in DMF in the presence of triethylamine or DMF/water mixtures in the presence of NaHCO.sub.3 at a temperature in the range ambient to 90.degree. for 1-20 hours. The product was purified on an octadecylsilane column and the following compounds were thus prepared.
__________________________________________________________________________ ##STR11##Example R.sup.1 R.sup.2 Yield % Footnotes__________________________________________________________________________ 7. C(CH.sub.3).sub.2 COOH ##STR12## 17 1, 2, 3 8. C(CH.sub.3).sub.2 COOH ##STR13## 7 4, 2, 5 9. C(CH.sub.3).sub.2 COOH ##STR14## 16 6, 7, 810. C(CH.sub.3).sub.2 COOH ##STR15## 26 9, 2, 10__________________________________________________________________________Example R.sup.1 R Yield % Footnotes__________________________________________________________________________11 CH.sub.3 ##STR16## 33 11, 12, 1312 CH.sub.3 ##STR17## 36 14, 1513 C(CH.sub.3).sub.2 COOH ##STR18## 22 16, 17, 1814 C(CH.sub.3).sub.2 COOH ##STR19## 20 19, 17, 20__________________________________________________________________________Footnotes1. The starting material was prepared by reaction of 3-chloropyridazinewith trimethyloxonium tetrafluoroborate in CH.sub.2 Cl.sub.2 for 18 hoursto give3-chloro-1-methylpyridazinium tetrafluoroborate.2. HPLC eluant MeOH/water/HOAc 20:79:1 v/v/v.3. n.m.r. in solvent B: 1.53(s, 3H); 1.52(s, 3H); 3.56(m, 2H); 4.26(s,3H); 4.1(d,1H); 4.6(d, 1H); 5.17(d, 1H); 5.88(d, 1H); 6.94(s, 1H); 7.7(d, 1H);7.98(dd,1H); 8.95(d, 1H).4. The starting material was prepared by reaction of 3-fluoropyrazinewithtrimethyloxonium tetrafluoroborate in CH.sub.2 Cl.sub.2 for 4 hoursunder reflux to give 1-methyl-3-fluoropyrazinium tetrafluoroborate.5. n.m.r. in solvent B: 1.53(s, 3H); 1.54(s, 3H); 3.55(m, 2H); 4.21(s,3H);4.2(d, 1H); 4.7(d, 1H); 5.15(d, 1H); 5.85(d, 1H); 7.0(s, 1H); 8.0(d, 1H); 8.19(s, 1H); 8.6(d, 1H).6. The starting material was obtained as follows. Reaction of 3,6-dichloropyridazine with trimethyloxonium tetrafluoroborate gave3,6-dichloro-1-methylpyridazinium tetrafluoroborate. To this compound(2.0 mmole) in acetonitrile was added NaHCO.sub.3 (2.0 mmole), a fewdrops of triethylamine and 2-acetamido-1-mercaptoethane (2.4 mmole).After 1 hour at ambient temperature the mixture was filtered,the filtrate evaporated and ether and CH.sub.2 Cl.sub.2 added. Theproductcrystallised to give 3-chloro-1-methyl-6-(2-acetamidoethylthio)pyridaziniutetrafluoroborate.7. HPLC eluant MeOH/water/HOAc 23:76:1 v/v/v.8. n.m.r. in solvent B: 1.53(s, 1H); 1.54(s, 1H); 3.6(br s, 2H); 3.99(s,3H);4.6(d, 1H); 4.8(d, 1H); 5.17(d, 1H); 5.9(d, 1H); 6.95(s, 1H); 8.16(dd,2H).9. The starting material was obtained by reaction of 4-methoxypyridazinewith trimethyloxonium tetrafluoroborate in CH.sub.2 Cl.sub.2 for 8 hoursatambient temperature to give 4-methoxy-1-methylpyridaziniumtetrafluoroborate.10. n.m.r in solvent B: 1.53(br s, 6H); 4.1(s, 3H); 4.39(m, 2H); 5.19(d,1H);5.85(d, 1H); 7.0(s, 1H); 7.37(dd, 1H); 8.55(d, 1H); 8.9(d, 1H).11. The 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.-- Z)-methoxyimino)acetamido]ceph-3-em-4-carboxylic acid trifluoroacetate used as startingmaterial may be prepared as follows: To a solution of cefotaxime (5.24g.)in phosphate buffer (pH 6.4, 440 ml.) was added sodium azide (2.86 g.)andsodium iodide (1.65 g.) and the mixture was immersed in a 70.degree. bathwithstirring for 4.5 hours. The solvent was evaporated to the point ofprecipita-tion and then the pH adjusted to 2.5 with 2N aqueous HCl. The resultingprecipitate was collected, washed with water, acetone and ether anddriedover P.sub.2 O.sub.5 to give 3-azidomethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.-- Z)-methoxyimino)acetamido]ceph-3-em-4-carboxylic acid in quantative yield,having the following n.m.r. in solvent A: 3.4(d, 1H); 3.7(d, 1H);3.86(s,3H); 3.95(d, 1H); 4.4(d, 1H); 5.15(d, 1H); 5.78(d, 1H); 6.75(s, 1H).To a stirred suspension of Raney nickel (16 g.) in MeOH (13 ml.) at0.degree. wasadded a solution of 3-azidomethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.--)-methoxyimino)acetamido]ceph-3-em-4-carboxylic acid (2.96 g.) in MeOH/TFA (14 ml., 1.13 ml.). After effervescence ceased the mixture wasdilutedwith MeOH and filtered through paper. The filtrate was evaporated, theresidue purified by HPLC using water/HOAc/MeOH 79:1:20 v/v/vas eluant and the product dried over P.sub.2 O.sub.5 to give3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.-- Z)-methoxyimino)acetamido]ceph-3-em-4-carboxylic acid trifluoroacetate(yield 45%) having the following n.m.r. in solvent A: 3.5-4.2(m, 4H);3.9(s, 3H); 5.15(d, 1H); 5.85(d, 1H); 6.75(s, 1H).12. HPLC eluant MeOH/water/HOAc 10:89:1 v/v/v.13. n.m.r. in solvent B: 3.46(d, 1H), 3.72(d, 1H); 3.97(s, 3H); 4.25(s,3H);4.11(d, 1H); 4.63(d, 1H); 5.13(d, 1H); 5.73(d, 1H); 6.97(s, 1H); 7.57(d, 1H); 7.98(dd, 1H); 8.93(d, 1H).14. HPLC eluant MeOH/water/HOAc 15:84:1 v/v/v.15. n.m.r. in solvent B: 3.43(d, 1H); 3.69(d, 1H); 3.97(s, 3H); 4.19(s,3H);4.23(d, 1H); 4.66(d, 1H); 5.12(d, 1H); 5.69(d, 1H); 7.0(s, 1H); 7.98(d, 1H); 8.2(s, 1H); 8.6(d, 1H).16. The starting material was prepared by reaction of 3,6-difluoropyridazineand trimethyloxonium tetrafluoroborate in CH.sub.2 Cl.sub.2 for 15 hoursat ambienttemperature to give 3,6-difluoro-1-methylpyridazinium tetrafluoroborate.This compound in acetonitrile was treated with a solution of ammonia inchloroform in the presence of 1 equivalent of NaHCO.sub.3 for 0.5 hoursat ambient temperature to give 3-amino-6-fluoro-2-methyl pyridaziniumtetrafluoroborate.17. HPLC eluant MeOH/water/HOAc 25:74:1 v/v/v.18. n.m.r. in solvent B: 1.53(s, 6H); 3.5(m, 2H); 3.7(s, 3H); 3.95(d,1H); 4.5(d,1H); 5.15(d, 1H); 5.8(d, 1H); 7.0(s, 1H); 7.3(s, 2H).19. The starting material was prepared by reaction of 2,5-dimethyl-3-fluoro-pyrazine and trimethyloxonium tetrafluoroborate in CH.sub.2 Cl.sub.2 atambienttemperature for 5 hours to give 3-fluoro-1,2,5-trimethylpyraziniumtetrafluoroborate.20. n.m.r. in solvent B: 1.53(s, 6H); 2.38(s, 3H); 2.53(s, 3H); 3.5(m,2H); 4.1(s,3H); 4.2(d, 1H); 4.85(d, 1H); 5.1(d, 1H); 5.85(d, 1H); 7.05(s, 1H);7.9(s, 1H).
EXAMPLES 15-16
The general process described in Examples 7-14 was repeated using the appropriate 3-aminomethylcephalosporin derivative and the appropriate chloroheterocycle and the following compounds were obtained.
__________________________________________________________________________ ##STR20##Example R Yield % Footnotes__________________________________________________________________________15 ##STR21## 35 116 CH.sub.2 CH.sub.2 Br 11 2__________________________________________________________________________Footnotes1. n.m.r. in solvent A: 2.68(s, 3H); 3.6(m, 4H); 4.0-4.8(m, 4H); 4.25(s,3H);5.15(d, 1H); 5.81(d, 1H); 7.0(s, 1H); 7.61(d, 1H); 7.99(dd, 1H); 8.97(d,1H).2. n.m.r. in solvent A: 3.6-3.9(m, 4H); 4.0-4.6(m, 4H); 4.26(s, 3H);5.14(d, 1H);5.82(d, 1H); 6.97(s, 1H); 7.65(d, 1H); 8.02(dd, 1H); 9.0(d,__________________________________________________________________________1H).
The starting materials for use in the above process may be obtained as follows:
Reaction of ethyl 2-(2-tritylaminothiazol-4-yl)-2-((Z)-2-bromoethoxyimino)acetate (UK patent application 2017702A) with 2-mercapto-5-methyl-1,3,4-thiadiazole and hydrolysis of the ester gave 2-(2-tritylaminothiazol-4-yl)-2-[(Z)-2-(5-methyl-1,3,4-thiadiazol-2-ylthio)ethoxyimino]acetic acid; n.m.r. in solvent C: 2.7 (s,3H); 3.65 (t,2H); 4.51 (t,2H); 6.65 (s,1H); 7.34 (s,15H).
Triethylamine (1.0 mmole.) and phosphorus pentachloride (1.0 mmole) were added to a solution of 2-(2-tritylaminothiazol-4-yl)-2-[(Z)-2-(S-methyl-1,3,4-thiadiazol-2-ylthio)ethoxyimino]acetic acid (1.0 mmole) in CH.sub.2 Cl.sub.2 (2.5 ml.) under argon at 0.degree. and the mixture stirred for 1.5 hours. The solvent was evaporated and the residue dissolved in CH.sub.2 Cl.sub.2. To this solution was added a solution of 7-amino-3-azidomethylceph-3-em-4-carboxylic acid (1.0 mmole.) in CH.sub.2 Cl.sub.2 (2.5 ml.) under argon [this solution having been previously treated at 0.degree. with N,O-bistrimethylsilylacetamide (2.0 mmole.) and allowed to warm to ambient temperature over 2 hours]. After 1.5 hours the mixture was diluted with CH.sub.2 Cl.sub.2 and the organic layer washed with water, brine and dried (MgSO.sub.4). Evaportion of the solvent and purification of the residue by chromatography on silica gel gave the product. Using this general process the following compounds were prepared.
__________________________________________________________________________ ##STR22##R Footnotes__________________________________________________________________________ ##STR23## 1CH.sub.2 CH.sub.2 Br 2, 3__________________________________________________________________________Footnotes1. n.m.r. in solvent A: 2.68(s, 3H); 3.64(m, 4H); 3.92(d, 1H); 4.46(d,1H);4.48(m, 2H); 5.2(d, 1H); 5.8(d, 1H); 6.92(s, 1H); 7.37(s, 15H).2. The starting material was 2-(2-tritylamino-thiazol-4-yl)-2-(( .sub.--Z)-2-bromoethoxyimino)acetic acid (UK patent application 2017702A).3. n.m.r. in solvent A: 3.5-3.9(m, 4H); 4.2-4.5(m, 4H); 5.16(d, 1H);5.76(d, 1H);6.8(s, 1H); 7.34(s, 15H).
The 3-azidomethylcephalosporin derivative was dissolved in formic acid and treated with an excess of wet Raney nickel for 50 minutes. The mixture was filtered through diatomaceous earth and the pad rinsed with MeOH/water 1:1 v/v. The filtrate was evaporated and the residue dissolved in TFA/water 9:1 v/v (5 ml.) at ambient temperature. After 1.5 hours the solvent was evaporated and the residue purified by chromatography on Diaion HP 20 resin, eluting with increasing proportions of MeOH in water. There were thus obtained the following compounds.
______________________________________ ##STR24##R Footnotes______________________________________ ##STR25## 1CH.sub.2 BH.sub.2 Br 2______________________________________Footnotes1. n.m.r. in solvent A: 2.67(s, 3H); 3.62(m, 6H); 4.47(t, 2H); 5.15(d, 1H); 5.86(d, 1H); 7.0(s, 1H).2. n.m.r. in solvent A: 3.65-3.9(m, 6H); 4.42(t, 2H); 5.17(d, 1H); 5.88(d, 1H); 6.97(s, 1H).
EXAMPLE 17
To a solution of 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid (400 mg.) and 4-chloro-1,7-dimethyl-1,8-naphthyridinium tetrafluoroborate (190 mg.) in DMF (2 ml.) was added triethylamine (285 ul.) at ambient temperature. After 1 hour the solvent was evaporated and the residue purified by HPLC on an octadecylsilane column using MeOH/aqueous ammonium carbonate (2 g./l) 30:70 v/v as eluant to give the following compound ##STR26## in 10% yield, n.m.r. in solvent B: 1.5 (s,6H); 2.73 (s,3H); 3.6 (s,2H); 4.1 (s,3H); 4.65 (s,2H), 5.2 (d,1H); 6.9 (d,1H), 7.0 (s,1H); 7.0 (d,1H); 7.7 (d,1H); 8.8 (dd,1H).
The starting material may be obtained as follows. 3-Carboxy-4-hydroxy-7-methyl-1,8-naphthyridine was heated at 300.degree. in a tube equipped with a cold finger to give 4-hydroxy-7-methyl-1,8-naphthyridine. Reaction of this compound with phosphorous oxychloride at 100.degree. for 30 minutes gave 4-chloro-7-methyl-1,8-naphthyridine. Reaction of this compound with trimethyloxonium tetrafluoroborate in CH.sub.2 Cl.sub.2 at ambient temperature for 3 hours gave, after evaporation of solvent and washing of the residue with ether, 4-chloro-1,7-dimethyl-1,8-naphthyridinium tetrafluoroborate which was used without further purification.
EXAMPLES 18-24
The following compounds were prepared by reaction of 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid with the appropriate quaternary heterocycle in the manner described in Example 1.
______________________________________ ##STR27##Exam- Yield Foot-ple R (%) notes______________________________________ ##STR28## 22 1 ##STR29## 27 2, 320. ##STR30## 40 4, 5 ##STR31## 14 6, 7 ##STR32## 13 8, 9 ##STR33## 14 10, 11 ##STR34## 16 12, 13______________________________________Footnotes1. N.m.r. in solvent A: 1.42(s, 6H), 2.52(s, 3H), 2.7(s, 3H), 3.68(dd,2H), 4.42(d, 1H), 4.8(d, 1H) 5.12(d, 1H), 5.9(d, 1H), 7.0(s, 1H),7.82(m, 1H), 8.1(d, 1H).2. N.m.r. in solvent A: 1.42(s, 6H), 3.32(d, 1H), 3.61(d, 1H),4.22(d, 1H), 4.61(d, 1H), 5.05(d, 1H), 5.76(d, 1H), 6.72(s,1H) 7.02(d, 1H), 7.78(d, 1H), 8.1(d, 1H), 8.7(d, 1H).3. The staring material may be prepared as follows: Lawesson'sreagent (5.26 g.) was added to a boiling solution of8H-thiazolo[3,2-a]pyrimidin-7-one (4.0 g.) in CH.sub.3 CN(200 ml.) and the mixture was heated under reflux with stirringfor 1.5 h. On cooling silica gel (70-230 mesh, 15 g.) was addedand the mixture was evaporated to dryness. The residue wasapplied to a column of silica gel (300 g.) and eluted withMeOH/CH.sub.2 Cl.sub.2 (5-10%) to give 8H-thiazolo[3,2-a]-pyrimidin-7-thione (2.7 g.). Trimethyloxonium tetrafluoroborate(2.0 g.) was added to a stirred suspension of 8H-thiazolo[3,2-a]-pyrimidin-7-thione (1.5 g.) in dry CH.sub.3 CN (50 ml.). Afterstirring for 2 h, the solution was concentrated to about 20 ml.,and with vigorous stirring anhydrous ether (100 ml.) was addedto precipitate 7-methylthiothiazolo[3,2-a]pyrimidiniumtetrafluoroborate (1.3 g.), having nmr in d.sub.6 DMSO: 2.78(s, 3H),8.04(d, 1H), 8.22(d, 1H), 8.55(d, 1H), 9.24(d, 1H).4. N.m.r. in solvent A: 1.42(s, 6H), 3.5(d, 2H) 3.68(m, 2H), 4.18(d,1H), 4.6(m, 3H), 5.1(d, 1H), 5.82(d, 1H), 6.62(d, 1H), 6.72(s, 1H),8.12(d, 1H).5. Starting from 2,3-dihydro-8H-thiazolo[3,2-a]pyrimidin-7-one,2,3-dihydro-7-methylthiothiazolo[3,2-a]pyrimidiniumtetrafluoroborate was prepared in the same way as described infootnote 3.6. N.m.r. in solvent A: 1.42(s, 6H), 3.58(dd, 2H), 4.25(d, 1H),4.75(d, 1H), 5.1(d, 1H), 5.82(d, 1H), 6.75(s, 1H), 7.39(s, 1H),7.6(d, 1H), 9.21(s, 1H).7. Starting from 5-methoxycarbonyl-8H-thiazolo[3,2-a]-pyrimidin-7-one, 5-methoxycarbonyl-7-methylthiothiazolo[3,2-a]-pyrimidinium tetraborate was prepared in the same way asdescribed in footnote 3. Upon reaction with the amino-methylcephalosporin, as described in Example 1, this materialunderwent hydrolysis of the methyl ester in addition todisplacement of the methylthio group.8. N.m.r. in solvent A: 1.12(t, 3H), 1.42(s, 6H), 2.7(s, 2H),3.3(d, 1H), 3.58(d, 1H), 4.2(d, 1H), 4.6(d, 1H), 5.09(d, 1H),5.68(d, 1H), 6.66(s, 1H), 8.0(d, 1H), 8.1(s, 1H), 9.18(d, 1H).9. The starting material may be prepared as follows:2-Amino-5-ethoxycarbonylmethylthiazole (3.72 g.), ethylpropiolate (1.96 g.) and 25% aqueous tetramethylammoniumhydroxide (0.15 ml.) in absolute EtOH (50 ml). were heated underreflux for 16 h. The solution was concentrated and the residuepurified by column chromatography (silica gel; 10%MeOH/CH.sub.2 Cl.sub.2) to give 3-ethoxycarbonylmethyl-8H-thiazolo[3,2-a]pyrimidin-7-one (1.6 g.), as a solid havingnmr in d.sub.6 DMSO: 1.2(t, 3H), 4.1(m, 4H), 6.15(d, 1H),7.15(s, 1H), 8.18(d, 1H). This material was converted to3-ethoxycarbonylmethyl-7-methylthiothiazolo[3,2-a]pyrimidiniumtetrafluoroborate in the same way as described in footnote 3.10. N.m.r. in solvent A: 1.4(s, 6H), 3.35(d, 1H), 3.65(d, 1H),4.32(d, 1H), 4.65(d, 1H), 5.05(d, 1H), 5.78(d, 1H), 6.75(s, 1H),7.12(d, 1H), 7.7(m, 2H), 8.25(m 1H), 9.23(d, 1H).11. 1H-Benzo[4,5]thiazolo[3,2-a]pyrimidin-2-one was converted to2-methylthiobenzo[4,5]thiazolo[3,2-a]pyrimidinium tetra-fluoroborate in the same was as described in footnote 3.12. N.m.r. in solvent A: 1.4(s, 6H), 2.75(s, 3H) 3.35(d, 1H), 3.61(d,1H), 4.28(d, 1H) 4.68(d, 1H), 5.04(d, 1H). 5.78(d, 1H), 6.72(s, 1H),7.03(d, 1H), 8.98(d, 1H).13. 8H-2-Methyl-1,3,4-thiadiazolo[3,2-a]pyrimidin-7-one wasconverted to 2-methyl-7-methylthio-1,3,4-thiadiazolo[3,2-a]-pyrimidinium tetrafluoroborate in the same way as described infootnote 3.
EXAMPLES 25-28
To a stirred suspension of 3-aminomethyl-7-[2-(2 aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4 carboxylic acid (200 mg.-0,4 mmole) and triethylamine (120 .mu.l. - 0.8 mmole) in DMF(2 ml.) at 25.degree. C. was added to the appropriate N-alkylated-4-methylthio pyrimidinium salt (0.6 mmole).
After 30 minutes, the solution was evaporated to dryness under reduced pressure and the residue purified by preparative HPLC (octadecyl silane column).
Using this general process and the appropriate quaternised heterocycle the following compounds were obtained.
__________________________________________________________________________ ##STR35##Examples R Yield (%) Footnotes__________________________________________________________________________ ##STR36## 67 1 2 ##STR37## 25 3 4 ##STR38## 24 5 6 ##STR39## 25 7 8__________________________________________________________________________Footnotes1. N.m.r. in solvent B: 1.54(s, 6H), 3.60 (m, 2H), 4.3(d, 1H), 4.8(d,1H), 5.2(br, 1H),5.3(s, 2H), 5.9(br, 1H), 7.0(d, 1H), 7.1(s, 1H), 7.5(s, 1H), 8.3(s, 1H),9.0(s, 1H).2. The pyrimidinium salt may be prepared as follows: A suspension of4-methylthio-pyrimidine (350 mg. 2.8 mmole) and alkyl halide (2.8 mmole) was heated at80.degree. C.for 1 hour. The residue was triturated with EtOAc and the resulting solidfilteredoff. N.m.r. in solvent B: 2.52(s, 3H), 5.6(s, 2H), 7.5(s, 1H), 8.1(d,1H), 9.0(d, 1H),9.62(s, 1H).3. N.m.r. in solvent B: 1.54(s, 6H), 3.6(m, 2H), 4.3(d, 1H), 4.8(d, 1H),5.0(d, 2H);5.2(d, 1H), 5.9(d, 1H), 6.8(s, 1H), 7.05(m, 3H), 7.45(m, 5H), 8.3(d, 1H),8.95(s, 1H).4. The starting material may be prepared in the same way as described infootnote2. N.m.r in solvent B: 2.74(s, 3H), 5.36(d, 2H), 6.8(m, 2H), 7.4(m, 5H),8.12(d, 1H),8.93(d, 1H), 9.6(s, 1H).5. N.m.r. in solvent B: 1.54(s, 6H), 3.56(m, 2H), 4.3(d, 1H), 4.8(d, 1H),5.2(d, 1H),5.6(s, 2H), 5.9(d, 1H) 7.0(m, 2H), 7.05(s, 1H), 7.24(s, 1H), 7.46(d, 1H),8.2(d, 1H),9.0(s, 1H).6. The starting material may be prepared in the same was as described infootnote 2.N.m.r. in solvent B: 2.74(s, 3H), 5.87(s, 2H), 6.9-7.8(m, 3H), 8.06(d,1H),8.95(d, 1H), 9.6(s, 1H).7. N.m.r. in solvent B: 1.54(s, 6H), 3.6(m, 2H), 4.4(d, 1H), 4.8(d, 1H),5.2(d, 1H),5.8(s, 2H), 5.95(d, 1H), 7.04(d, 1H), 7.15(s, 1H), 7.7(s, 2H), 8.3(d,1H), 9.0(s, 1H).8. The starting material may be prepared in the same way as described infootnote2. N.m.r. in solvent B: 2.74(s, 3H), 6.12(s, 2H), 7.7(s, 2H), 8.3(d,1H),9.1(d, 1H), 9.7(s, 1H).
EXAMPLE 29 ##STR40##
To a stirred suspension of 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-methoxyimino)acetamido]ceph-3-em-4-carboxylic acid (205 mg.-0.5 mmole) and triethylamine (70 .mu.l. 0.5 mmole) in DMF (2 ml.) at 25.degree. C. was added the quaternised pyrimidine described in Example 28, footnote 8 (185 mg. 0.5 mmole). After 30 min the solution was evaporated to dryness and the residue was purified by preparative HPLC (octecycl silane column). Yield 33%. N.m.r. in solvent B: 3.6 (m, 2H), 4.0 (s, 3H), 4.3 (d, 1H), 4.8 (d, 1H), 5.15 (m, 1H), 5.85 (br, 3H), 7.05 (m, 2H), 7.7 (s, 2H), 8.3 (d, 2H), 9.0 (s, 1H).
EXAMPLE 30 ##STR41##
To a stirred suspension of 3-ethylaminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid (160 mg. 0.31 mmole) and triethylamine (85 .mu.l - 0.62 mmole) in DMF (2 mi.) at 25.degree. C. was added the N-alkylated-4-methylthiopyrimidine (140 mg. 0.62 mmole).
To a stirred suspension of 3-ethylaminomethyl-7-[92-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid (160 mg. 0.31 mmole) and triethylamine (85 .mu.l - 0.62 mmole) in DMF (2 ml.) at 25.degree. C. was added the N-alkylated-4-methylthiopyrimidine (140 mg. 0.62 mmole). After 2 hr. at 45.degree. C., the solution was evaporated to dryness and the residue was purified by preparative HPLC.
N.m.r. in solvent B: 1.16 (m, 3H), 1.54 (s, 6H), 3.6 (m, 4H), 4,6-5.3 (m, 5H), 5.9 (d, 1H), 7.05 (s, 1H), 7.2 (m, 1H), 8.3 (d, 1H), 8,8 (s, 1H).
The starting material may be prepared as follows:
A suspension of 4-methylthiopyrimidine (504 mg. 4 mmoles) and 2-chloroacetamide (375 mg. 4 mmoles) was heated at 90.degree. C. for 3 hr. The reaction mixture was triturated with EtOAc, and the resulting solid was filtered off.
N.m.r. in solvent B: 2.52 (s, 3H), 5.2 (s, 2H), 8.1 (d, 1H), 8.8 (d, 1H), 9.3 (s, 1H).
EXAMPLES 31-32
The process described in Example 29 was repeated, using 3-ethylaminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-2-chloroethoxyimino)acetamido]ceph-3-em-4-carboxylic acid as starting material, to give the following compounds:
______________________________________ ##STR42##Example R Yield (%) Footnotes______________________________________ ##STR43## 12 1, 2, 3 ##STR44## 31 1, 2, 4______________________________________Footnotes1. To a solution of 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-2-chloroethoxyimino)acetamido]ceph-3-em-4-carboyxlic acid.(2 g. 4.3 mmoles) in MeOH (120 ml.) was added triethylamine(0.6 ml. 4.3 mmoles) and sodium cyanoborohydride (0.14 g.2.2 mmoles). After addition of acetaldehyde (0.15 ml. 2.2mmoles) the mixture was stirred at room temperature for 1 hour.After evaporation the residue was purified on HP20 resin. N.m.r.in solvent B: 1.21(s, 3H), 2.8-3.1(m, 2H), 3.2-4.2(m, 6H),4.2-4.6(m, 2H), 5.4(d, 1H), 5.8(d, 1H), 7.1(s, 1H).2. The above 3-ethylaminomethyl cephalosporin was reacted withthe appropriate quaternised heterocycle in the same way asdescribed in Example 30.3. N.m.r. in solvent B: 1.2(br, 3H), 3.5(br, 4H), 3.85(s, 3H),4.0(br, 2H), 4.5(m, 4H), 5.3(d, 1H), 5.9(d, 1H), 7.05(s, 1H), 7.4(d,1H), 7.8(d, 1H).4. N.m.r. in solvent B: 1.2(br, 3H), 3.0-4.0(m, 6H), 4.2-5.3(m, 7H),5.85(d, 1H), 7.04(s, 1H), 7.2(d, 1H), 8.3(d, 1H) 8.7(s, 1H).
EXAMPLES 33-34
The process described in Example 25 was repeated, using the appropriate methylthio heterocycle, to give the following compounds.
______________________________________ ##STR45##Example R Yield (%) Footnotes______________________________________ ##STR46## 42% 1, 2 ##STR47## 14% 3, 4______________________________________1. N.m.r. in solvent B: 1.55(s, 6H), 3.6(m, 2H), 4.02(s, 3H),4.7(d, 1H), 5.1(d, 1H), 5.2(d, 1H), 5.9(d, 1H), 7.05(s, 1H),7.9(m, 1H), 9.1(m, 3H).2. The starting material may be obtained as follows: 4-Methylthio-pyrido[2,3-d]pyrimidine (700 mg. 4 mmoles) was reacted withtrimethyloxonium tetrafluoroborate (582 mg. 4 mmoles) inCH.sub.2 Cl.sub.2 (20 ml.) at ambient temperature for 1 hr. Afterfiltration the solvent was evaporated to give a solid which wasused without further purification.3. N.m.r. in solvent B: 1.55(s, 6H), 3.65(br, 2H), 4.37(s, 3H), 4.6(d,1H), 5.0(d, 1H), 5.2(d, 1H), 5.9(d, 1H), 7.0(s, 1H), 8.0(m, 1H),8.8(s, 1H), 9.45(m, 2H).4. The solid obtained in footnote 2 was used after triturationin MeOH.
EXAMPLE 35 ##STR48##
To a stirred suspension of 3-aminomethyl-7-[2-(2 aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid (200 mg.-0.4 mmole) and triethylamine (120 .mu.l. - 0.8 mmole) in DMF (2 ml.) at 25.degree. C. was added the N-alkylated-4-methylthio heterocycle (0.6 mmole). After 30 min., the solution was evaporated to dryness under reduced pressure and the residue purified by preparative HPLC (octadecyl silane column).
N.m.r. in solvent B: 1.54 (s, 6H), 3.6 (m, 2H), 4.35 (d, 1H), 4.8 (d, 1H), 5.2 (d, 1H), 5.8 (s, 2H), 5.9 (d, 1H), 7.0 (m, 4H), 7.9 (d, 2H), 8.1 (d, 1H), 8.7 (s, 1H).
The methylthio heterocycle used as starting material may be prepared in the same way as described in Example 25, footnote 2.
N.m.r. in solvent B: 2.75 (s, 3H), 6.14 (s, 2H), 7.0 (d, 2H), 8.0 (d, 2H), 8.2 (d, 1H), 8.8 (d, 1H), 9.4 (s, 1H).
EXAMPLES 36-37
The process described in Example 30 was repeated, using the appropriate methylthio heterocycle as starting material, to give the following compounds:
______________________________________ ##STR49##Exam-ple R Yield (%) Footnotes______________________________________ ##STR50## 46% 1, 2 ##STR51## 46% 3, 4______________________________________Footnotes:N.m.r. solvent B: 1.52(s, 9H), 3.51(s, 2H), 4.0(s, 5H), 4.95(d, 1H),5.2(m, 2H), 5.9(d, 1H), 7.0(s, 1H), 7.9(m, 1H), 8.7(d, 1H), 9.1(m, 2H).2. The starting material was described in Example 33, footnote 2.3. N.m.r in solvent B: 1.13(m, 3H), 1.53(s, 6H), 3.2-3.9(m, 4H),4.6(d, 1H), 4.8-5.3(m, 2H), 5.6(s, 2H), 5.85(d, 1H), 6.9-7.6(m,5H), 8.4(d, 1H), 9.0(s, 1H).4. Starting material described in Example 27, Footnote 6.
EXAMPLE 38 ##STR52##
The general process described in Example 27 was repeated, using the appropriate 4-methylsulphinyl pyridinium salt.
N.m.r. in solvent B: 1.54 (s, 6H), 2.5 (br, 2H), 2.8 (br, 2H), 3.55 (m, 2H), 4.35 (br, 2H), 5.2 (d, 1H), 5.7 (m, 1H), 5.9 (d, 1H), 7.0 (m, 4H), 8.2 (m, 2H).
The pyridinium salt was used as starting material may be obtained by condensation of the known 4-methylthiopyridine and 3-bromo-1-cyanocyclopentene in MeOH at 20.degree. C. for 12 hr.
N.m.r. in solvent B: 2,3-3.0 (m, 7H), 5.8 (br, 1H), 6.7 (s, 1H), 7.6 (d, 2H), 8.5 (d, 2H).
This product, as its tetrafluoroborate salt, was oxidised with MCPBA in CH.sub.2 Cl.sub.2 /TFA at 20.degree. C. for 3 hr. After evaporation of the solvents the solid was triturated with ether and used without further purification.
EXAMPLES 39-53
A solution of NaHCO.sub.3 (0.6 mM) in water (750 ml.) was added to a solution of 7-acyl-3-aminomethyl- cephalosporin derivative (0.2 mM) in DMF (2 ml.), followed after a few minutes by the appropriate quaternised heterocyclic starting material (0.22 mM). The mixture was stirred at room temperature during 3hrs. The mixture was evaporated to dryness and the residue purified by HPLC on an octadecylsilane column.
Using this general process the following compounds were prepared.
__________________________________________________________________________ ##STR53## YieldExample R.sub.1 R.sub.2 (%) Footnotes__________________________________________________________________________39. CH.sub.2 CHCHCO.sub.2 H ##STR54## 36 1, 240. CH.sub.2 CH.sub.2 SOCH.sub.3 ##STR55## 32 3, 441. CH.sub.2 CH.sub.2 SO.sub.2 CH.sub.3 ##STR56## 59 5, 642. CH.sub.2 CH.sub.2 SOCH.sub.3 ##STR57## 32 7, 843. CH.sub.2 CN ##STR58## 37 9, 1044. CH.sub.2 CN ##STR59## 49 11, 1245. CH.sub.2 CH.sub.2 F ##STR60## 40 13, 14 ##STR61## ##STR62## 37 15, 16 ##STR63## ##STR64## 55 17, 1848. CH.sub.2 CH.sub.2 F ##STR65## 21 19, 2049. CH.sub.2 CF.sub.3 ##STR66## 4 21, 2250. CH.sub.2 CF.sub.3 ##STR67## 48 23, 2451. CH.sub.2 CF.sub.3 ##STR68## 31 25, 26 ##STR69## ##STR70## 30 27, 2853. CH.sub.2 CH.sub.2 F ##STR71## 52 29, 30__________________________________________________________________________1. Reaction time: 1 Hour 40 minutes.2. N.m.r. in solvent B: 3.35-3.85(m, 2H), 4.3(s, 3H), 4.1-4.5(m, 2H),4.8(d, 2H), 5.2(d, 1H),5.9(d, 1H), 6.1(d, 1H), 7.05(s, 1H), 6.8-7.2(m, 1H), 7.8-9.1(m, 4H).3. The process was carried out using 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-(Z-methylthio-ethoxyimino)acetamido]ceph-3-em-4-carboxylic acid and 3-chloro-1-methyl-pyridinium iodide,during 2 hours. The crude product was not purified after evaporation butwas oxidised directly asfollows: A solution of crude 7-[2-(2-aminothiazol-4-yl)-2-(Z-methylthioethoxyimino)acetamido]-3-(1-methyl-3-pyridazinio)aminomethylceph-3-em-4-carboxylic acid (0.5 mM)in TFA (4.0 ml.)and CH.sub.2 Cl.sub.2 (6.0 ml.) was cooled to -25.degree. C. withvigourous stirringm-chloroperbenzoic acid (0.5 mM) was added portion wise during 1 hr. Thetemperature wasallowed to rise to ambient, the mixture was evaporated to dryness and theresidue was purifiedby HPLC on an octadecylsilane column.4. N.m.r. in solvent B: 2.6(s, 3H), 3.2(d, 2H), 3.6(dd, 2H), 4.3(s, 3H),4.6(m, 4H), 5.2(d, 1H),5.8(d, 1H), 7.1(s, 1H), 7.65(d, 1H), 8.0(dd, 1H) 8.9(d, 1H).5. Reaction time: 1 Hour 45 minutes.6. N.m.r. in solvent B: 2.9(s, 3H), 3.5-3.7(m, 4H), 4.2(s, 3H),4.5-4.7(m, 4H), 5.1(d, 1H),5.57, (d, 1H), 7.05(s, 1H), 7.55(d, 1H), 8.0(dd, 1H), 9.0(d, 1H).7. Prepared in two stages, condensation and oxidatin as in footnote 3.Reaction time for thecondensation: 4 hours 30 minutes at 50.degree. C.8. N.m.r. in solvent B: 2.61(s, 3H), 3.2(d, 2H), 3.9(s, 5H), 4.7(m, 4H),5.2(d, 1H), 5.9(d, 1H),7.05(s, 1H), 7.65(d, 1H), 7.75(d, 1H).9. Reaction time: 2 hours 30 minutes at 50.degree. C.10. N.m.r. in solvent B: 3.53(m, 2H), 3.7(s, 3H), 4.23(m, 2H), 5.08(s,2H), 5.2(d, 1H), 5.77(d, 1H)7.08(s, 1H), 7.34(s, 2H).11. Reaction time: 2 hours.12. N.m.r. in solvent B: 3.51(m, 2H), 4.52(m, 2H), 5.05(s, 2H), 5.11(d,1H), 5.75(s, 2H), 5.8(d, 1H)7.0(d, 1H), 7.17(s, 1H), 7.78(s, 2H), 8.24(d, 1H), 8.96(s, 1H).13. Reaction temperature: 50.degree. C.14. N.m.r. in solvent B: 3.5(m, 2H), 3.7(s, 3H), 3.8-4.6(m, 5H), 4.9(m,1H), 5.15(d, 1H),5.78(d, 1H), 6.99(s, 1H), 7.3(s, 2H).15. Reaction temperature: 50.degree. C.16. N.m.r. in solvent B: 3.6(m, 2H), 3.72(s, 3H), 4.3(dd, 2H), 5.15(d,1H), 5.43(s, 2H),5.8(d, 1H), 7.06(s, 1H), 7.33(s, 2H), 7.73(s, 2H).17. Reaction time: 45 minutes at 50.degree. C.18. N.m.r. in solvent B: 3.55(m, 2H), 4.5(dd, 2H), 5.05(d, 1H), 5.45(s,2H), 5.8(d, 1H), 7.0(d, 1H),7.05(s, 1H), 7.65(d, 2H), 7.7(d, 1H), 8.05(d, 1H), 8.80(d, 1H).19. Reaction time: 20 hours.20 N.m.r. in solvent B: 3.5(m, 2H), 4.1-4.8(m, 5H), 4.9(m, 2H), 5.13(d,1H), 5.81(d, 1H),7.01(d, 1H), 7.02(s, 1H), 7.65(d, 1H), 8.11(d, 1H), 8.74(d, 1H).21. Reaction time: 1 hour.22. N.m.r. in solvent B: 3.42(d, 1H), 3.68(d, 1H), 4.07(s, 3H), 4.57(d,1H), 4.95(d, 1H), 4.64(d,1H), 4.82(d, 1H), 5.12(d, 1H), 5.77(d, 1H), 7.07(s, 1H), 7.91(s, 2H),8.85(s, 1H).23. Reaction time: 1 hour.24. N.m.r. in solvent B: 3.57(d, 1H), 3.65(d, 1H), 3.67(t, 2H), 4.18(d,1H), 4.4-4.9(m, 5H),5.11(d, 1H), 5.78(d, 1H), 6.61(d, 1H), 7.0(s, 1H), 8.15(d, 1H).25. Reaction time: 2 hours 30 minutes at 50.degree. C.26. N.m.r. in solvent B: 3.4(d, 1H), 3.62(d, 1H), 3.68(s, 3H), 3.95(d,1H), 4.42(d, 1H),4.63(d, 1H), 4.82(d, 1H), 5.11(d, 1H), 5.74(d, 1H), 7.0(s, 1H), 7.3(s,2H).27. Reaction time: 2 hours at 50.degree. C.28. N.m.r. in solvent B: 3.25(m, 2H), 3.6(s, 3H), 4.0(dd, 2H), 4.8(d,1H), 5.15(s, 2H), 5.5(d, 1H),6.65(s, 1H), 7.15(d, 1H), 7.4(d, 2H), 8.25(m, 3H).29. Reaction time: 20 hours.30. N.m.r. in solvent B: 3.55(m, 2H), 4.2-4.9(m, 5H), 4.98(t, 1H),5.19(d, 1H), 5.39(s, 2H),5.81(d, 1H), 7.02(s, 1H), 7.0-7.3(m, 2H), 7.38(d, 2H), 7.68(d, 2H),8.3-8.7(m, 2H).
The cephalosporin starting materials for use in the above process are obtained by the method described above.
Using this general process the following compounds were prepared.
______________________________________ ##STR72##R Footnotes______________________________________CH.sub.2 CHCHCO.sub.2 t-Bu 1, 2CH.sub.2 CH.sub.2 SCH.sub.3 3, 4CH.sub.2 CH.sub.2 SO.sub.2 CH.sub.3 5, 6CH.sub.2 CN 7CH.sub.2 CH.sub.2 F 8, 9 ##STR73## 10, 11, 12, 13CH.sub.2 CF.sub.2 14, 15, 16, 13 ##STR74## 17, 18, 15, 13______________________________________ Footnotes1. The starting material was prepared by reaction of 4-nitrobenzyl2-(2-tritylaminothiazol-4-yl)-2-(Z)-hydroxyiminoacetate with t-buyl4-bromobut-2-enoate and hydrogenolysis of the resulting ester togive 2-(2-tritylaminothiazol-4-yl)-2-[(Z)-3-t-butoxycarbonyl-prop-2-enyloxyimino]acetic acid. N.m.r. in solvent C: 1.44(s, 9H),4.75(d, 2H), 5.92(m, 1H), 6.56(s, 1H), 6.7-6.9(m, 2H), 7.29(s, 15H).2. N.m.r. in solvent C: 3.4(m, 2H), 3.9(d, 1H), 4.45(d, 1H), 4.9(m,2H), 5.02(d, 1H), 5.7(m, 1H), 5.95(d, 1H), 6.83(s, 1H), 6.9(d, 1H),7.32(s, 15H).3. The starting material was prepared by reaction of ethyl2-(2-tritylaminothiazol-4-yl)-2-((Z)-2-bromoethoxyimino)acetate(UK Patent Application 217702A) with the sodium salt of methylmercaptan and hydrolysis of the resulting ester to give 2-(2-trityl-aminothiazol-4-yl)-2-((Z)-2-methylthioethoxyimino)acetic acid. N.m.r.in solvent C: 2.1(s, 8H), 2.8(t, 2H) 4.3(t, 2H), 6.6(s, 1H), 7.3(s,15H).4. N.m.r. in solvent C: 2.1(s, 3H), 2.9(m, 2H), 3.5(s, 2H),3.95-4.6(m, 4H), 5.1(d, 1H), 5.95(d, 1H), 6.8(s, 1H), 7.35(s, 15H).5. The starting material was prepared by oxidation of the product inFootnote 3 to give 2-(2-tritylaminothiazol-4-yl)-2-((Z)-2-methanesulphonylethoxyimino)acetic acid.N.m.r. in solvent C: 2.9(s, 3H), 3.5(t, 2H),4.5(t, 2H), 6.65(s, 1H),7.3(s, 15H).6. N.m.r. in solvent C: 3.0(s, 3H, 3.5-3.7(m, 4H), 4.3(m, 2H), 4.7(m,2H),5.1(d, 1H), 5.9(d, 1H) 6.8(s, 1H), 7.4(s, 15H).7. N.m.r. in solvent A: 3.4(s, 2H), 3.9(d, 1H), 4.4(d, 1H), 4.8(s, 2H),4.9(d, 1H), 5.7(d, 1H), 6.8(s, 1H), 7.2(s, 15H).8. The starting material may be prepared as follows. Reaction of ethyl2-(2-tritylaminothiazol-4-yl)-2-[(Z)-2-hydroxyethoxyimino]acetate withdiethylamino sulphur trifluoride and hydrolysis of the resulting estergave 2-(2-tritylaminothiazol-4-yl)-2-[(Z)-2-fluoroethoxyimino]aceticacid.N.m.r. in solvent A: 4.15(m, 1H), 4.38(m, 1H), 4.48(m, 1H), 4.92(m, 1H),6.94(s, 1H), 7.4(s, 15H).9. N.m.r. in solvent C: 3.4(m, 2H), 4.2-4.6(m, 4H), 4.7(m, 1H), 5.0(m,1H), 5.1(d, 1H), 5.9(d, 1H), 6.9(s, 1H), 7.4(s, 15H).10. The starting material was prepared by reaction of 2-bromomethylimidazole with N-hydroxy-phthalimide, deprotection to give0-(imidazol-2-yl)methylhydroxylamine and condensation with2-(2-aminothiazol-4-yl)-glyoxylic acid to give 2-(2-aminothiazol-4-yl)-2-[(Z)-(imidazol-2-yl)-methyoxyimino]acetic acid. N.m.r. insolvent B: 5.42(s, 2H), 7.17(s, 1H), 7.67(s, 2H).11. A suspension of the product from Footnote 10 (4.4 mM) in DMF(40 ul.) at 0.degree. C. was dissolved by the addition of 3M HCl inether(8.8 mM). N-hydroxybenzotriazole (4.4 mM) and t-butyl 7-amino-3-azidomethylceph-3-em-4-carboxylate (4.0 mM) were added to thesolution, followed by dropwise addition of a solution of DCCI(4,4 mM) in CH.sub.3 CN (15 ml.) during 20 minutes. The mixture wasleft to stir for 23 hours at 0.degree. C. and 1 hour at roomtemperature.The precipitate was filtered, washed with DMF (3 ml.) and theresidue evaporated to dryness. Trituration at 0.degree. C. with aqueousNaHCO.sub.3 (5%) yielded a yellow solid which was treated at roomtemperature with TFA/H.sub.2 O (15:1) during 2 hours. Evaporation todyrness and purification of the residue by chromatography onDianion HP 20 resin gave the required product.12. N.m.r. in solvent B: 3.6(m, 2H), 4.2(dd, 2H), 5.25(d, 1H), 5.5(s,2H), 5.9(d, 1H), 7.25(s, 1H) 7.7(s, 2H).13. This molecule is not tritylated on the14. The starting material was prepared by condensation of0-(2,2,2-trifluoroethyl)hydroxylamine with 2-(2-aminothiazol-4-yl)-glyoxylic acid to give 2-(2-aminothiazol-4-yl)-2-[(Z)-2,2,2-trifluoro-ethoxyimino]acetic acid. N.m.r. in solvent B: 4.73(q, 2H), 6.93(s,1H).15. The method of condensation was that described in Footnote 11.16. N.m.r. in solvent B: 3.5(d, 1H), 3.75(d, 1H), 3.95(d, 1H),4.5(d, 1H), 4.75(d, 1H), 4.95(d, 1H), 5.3(d, 1H), 5.9(d, 1H), 7.2(s,1H).17. The starting material was prepared by reaction of0-(1-methylimidazol-2-yl)methylhydroxylamine with 2-(2-amino-thiazol-4-yl)glyoxylic acid to give 2-(2-aminothiazol-4-yl)-2-[(Z)-(1-methylimidazol-2-yl)methoxy-imino]acetic acid.N.m.r. in solvent B: 3.90(s, 3H), 5.51(s, 2H), 7.17(s, 1H),7.57(s, 1H), 7.62(s, 1H).18. N.m.r. in solvent B: 3.6(m, 2H), 3.9(s, 3H), 4.2(dd, 2H), 5.2(d,1H), 5.5(s, 2H), 5.85(d, 1H), 7.05(s, 1H), 7.7(dd, 2H).
Using the general process of reduction described in Footnote 3 of the description of starting materials for Example 16, the following compounds were prepared:
______________________________________ ##STR75##R Footnotes______________________________________CH.sub.2 CCHCO.sub.2 H 1CH.sub.2 CH.sub.2 SCH.sub.3 2CH.sub.2 CH.sub.2 SO.sub.2 CH.sub.3 3CH.sub.2 CN 4CH.sub.2 CH.sub.2 F 5 ##STR76## 6CH.sub.2 CF.sub.3 7 ##STR77## 8______________________________________1. N.m.r. in solvent B: 3.66(s, 2H), 3.78(m, 2H) 4.88(m, 2H),5.18(d, 1H), 5.88(d, 1H), 6.05(d, 1H), 6.92(d, 1H), 6.94(s, 1H).2. N.m.r. in solvent B: 2.1(s, 3H), 2.85(t, 2H), 3.6-3.9(m, 4H),4.4(t, 2H), 5.2(d, 1H), 5.9(d, 1H), 7.1(s, 1H).3. N.m.r. in solvent B: 2.95(s, 3H), 3.65(m, 6H), 4.6(t, 2H), 5.15(d,1H), 5.85(d, 1H), 7.05(s, 1H).4. N.m.r. in solvent A: 3.66(s, 2H), 3.78(q, 2H), 5.02(s, 2H),5.15(d, 1H), 5.84(d, 1H) 6.98(s, 1H).5. N.m.r. in solvent B: 3.6-4.0(m, 4H), 4.32(m, 1H), 4.4-4.7(m, 2H),5.08(m, 1H), 5.24(d, 1H) 5.58(d, 1H), 6.98(s, 1H).6. N.m.r. in solvent B: 3.8(m, 4H), 5.15(d, 1H), 5.43(s, 2H), 5.86(d,1H), 7.12, (s, 1H), 7.66(s, 2H).7. N.m.r. in solvent B: 3.4-4.0(m, 4H), 4.65(d, 1H), 4.85(d, 1H),5.17(d, 1H), 5.85(d, 1H), 7.05(s, 1H).8. N.m.r. in solvent B: 3.6-3.9(m, 4H), 3.9(s, 3H); 5.15(d, 1H),5.5(s, 2H), 5.85(d, 1H), 7.05(s, 1H), 7.65(dd, 2H).
EXAMPLES 54-75
The appropriate C3.sup.1 -N-substituted CAZAMCA (1 eq.) and an excess (1.5 eq.) of an appropriate quaternay heterocyclic salt were reacted in a mixture of DMF/H.sub.2 O (2:1) in the presence of NaHCO.sub.3 (4 eq.) at 40.degree. C. for 2.5 hr., and the product was purified by HPLC, using MeOH/H.sub.2 O/HOAc as eluant, to give the following compounds:
__________________________________________________________________________ ##STR78## YieldExample R R.sup.1 (%) Footnotes__________________________________________________________________________54 PhCH.sub.2 ##STR79## 30 1, 255 Et ##STR80## 58 3, 456. furfuryl ##STR81## 40 5, 657. CH.sub.2.CO.sub.2 H ##STR82## 31 7, 858. (CH.sub.2).sub.2 OH ##STR83## 14 9, 1059. Et ##STR84## 33 1160. Et ##STR85## 11 1261. Et ##STR86## 47 13 ##STR87## ##STR88## 10 14, 1563. (CH.sub.2).sub.2 OCH.sub.3 ##STR89## 36 16, 1764. Et ##STR90## 10 1865. i-Pr ##STR91## 5 19, 2066. Et ##STR92## 23 2167. (CH.sub.2).sub.2 NH.sub.2 ##STR93## 30 22, 2368. (CH.sub.2).sub.2 F ##STR94## 23 24, 2569. Et ##STR95## 15 2670. Me ##STR96## 22 27, 2871. Et ##STR97## 14 2972. allyl ##STR98## 13 30, 3173. Pr ##STR99## 18 3274. (CH.sub.2).sub.2 CN ##STR100## 33 33, 3475. (CH.sub.2).sub.2 NH.CHO ##STR101## 37 35__________________________________________________________________________Footnotes1. Eluant proportion 35:65:1. N.m.r. in solvent B: 1.55(m, 6H),3.4-3.6(m, 2H), 4.0(s,3H),4.7-5.0(m, 4H), 5.2(d, 1H), 5.9(d, 1H), 7.1(s, 1H), 7.2-7.5(m, 7H),8.4(d, 2H).2. The precursor C3.sup.1 N-benzyl CAZAMCA was obtained by reductiveamination ofbenzaldehyde with CAZAMCA. Thus, CAZAMCA (1 eq.) was treated withbenzaldehyde (1.4 eq) and Et.sub.3 N (1.4 eq.) in anhydrous MeOH in thepresence of 3Amolecular seives with NaBH.sub.3.CN (1.4 eq.) for 1/2 hr. at roomtemperature. Theproduct was purified by HPLC, eluant proportion 30:70:1. N.m.r. insolvent B: 1.5(s, 6H),3.5-3.7(m, 2H), 3.8-4.0(m, 2H), 4.0-4.3(m, 2H), 5.2(d, 1H), 5.9(d, 1H),7.0(s,1H), 7.4-7.6(m, 5H).3. Used 2 eq. pyridinium salt. Reaction time 1.5 hr. Eluent proportion25:75:1. N.m.r. insolvent B: 1.6(s, 6H), 3.2-3.8(m, 4H), 4.0(s, 3H), 4.6-4.8(m, 2H), 5.3(d,1H), 6.0(d, 1H),7.15(s, 1H), 7.2-7.4(m, 2H), 8.2-8.4(m, 2H).4. The precursor C.sub.3 ' N-ethyl CAZAMCA was obtained by reductiveamination ofacetaldehyde (1.1 eq.) with CAZAMCA (1.0 eq.) in presence of NaBH.sub.3CN (1.0 eq.) andEt.sub.3 N (1.0 eq.). The acetaldehyde was added dropwise at roomtemperature in MeOH, andthe mixture stirred for 1/2 hr. The product was purified by preparativeHPLC usingMeOH/H.sub.2 O/HOAc 10:90:1 v/v/v, followed by 15:85:1. N.m.r. in solventB: 1.3 (t, 3H):1.6(s, 6H), 2.8-3.2(m, 2H), (m, 2H) 3.7-4.1(m, 4H), 5.3(d, 1H), 6.0(d,1H), 7.2(s, 1H).5. N.m.r. in solvent B: 1.55(m, 6H), 3.3-3.6(m, 2H), 4.0(s, 3H),4.8-5.0(m, 4H), 5.2(d, 1H),5.95(d, 1H), 6.4-6.6(m, 2H), 7.1(s, 1H) 7.3-7.6(d, 2H), 7.6- 7.7(m, 1H)8.3-8.6(d, 2H).6. The precursor was obtained by reductive amination of 2-furaldehydewith CAZAMCA,as in Footnote 2. Reaction time 1.5 hr. Eluant proportion 20:80:1. N.m.r.in solvent B:1.6(s, 6H), 3.6-3.8(m, 2H), 3.9-4.1(m, 2H), 4.2-4.4(m, 2H), 5.25(d, 1H),5.95(d,1H), 6.5-6.8(m, 2H), 7.1(s, 1H), 7.8(d, 1H).7. Eluant proportion 30:70:1. N.m.r. in solvent B: 1.5-1.6(m, 6h),3.3-3.6(m, 2H),4.0(s, 3H), 4.4-4.6(m, 2H), 4.6-4.8(m, 2H), 5.3(d, 1H), 6.0(d, 1H),7.1(s, 1H),7.0-7.5(m, 2H), 8.3-8.5(m, 2H).8. Precursor obtained as in Footnote 4, starting from glyoxylic acid.Eluant proportion15:85:1. Yield 40% N.m.r. in solvent B; 1.6(m, 6H), 3.6-4.2(m, 6H),5.3(d, 1H),6.0(d, 1H), 7.1(s, 1H).9. Eluant proportion 20:80:1. N.m.r. in solvent B: 1.55(s, 6H),3.2-3.6(m, 2H),3.5-3.8(m, 4H), 3.35(s, 3H), 4.6-4.8(m, 2H), 5.2(d, 1H), 5.9(d, 1H),7.1(s, 1H),7.3(d, 2H), 8.3(d, 2H).10. Precursor obtained by reductive amination of glycolaldehyde as inFootnote 4. Eluantproportion 15:85:1. Yield 43%. N.m.r. in solvent B; 1.6(s, 6H),3.0-3.4(m, 2H),3.6-4.0(m, 4H), 4.2-4.6(m, 2H), 5.2(d, 1H), 5.9(d, 1H), 7.0(s, 1H).11. Reacted at 50.degree. C. for 2 hrs. Eluant proportion 30:70:1. N.m.r.in solvent B: 1.2(m,3H), 1.5(s, 6H), 3.2-3.8(m, 4H), 4.6-4.8(m, 2H), 5.2(s, 1H), 5.9(s, 1H),7.1(s, 1H),7.2-8.0(m, 7H).12. Reacted at 45.degree. C. for 20 hrs. Eluant proportion 25:75:1. N.m.rin solvent B:1.0-1.3(m, 3H), 1.55(s, 6H), 3.5-3.8(m, 4H), 3.8(s, 3H), 4.3(d, 1H),4.7(d, 1H),5.2(d, 1H), 5.8(d, 1H), 7.05(s, 1H), 7.35(d, 1H), 7.7(d, 1H).13. Reacted at room temperature for 3.5 hrs, using N-(Boc-aminoethyl)-4-chloropyridiniumchloride as the quaternary heterocyclic salt. The product was deprotectedby treating withTFA at room temperature for 1 hr. Eluant proportion 35:65:1. N.m.r. insolvent B:1.0-1.3(m, 3H), 1.6(s, 6H), 3.2-3.8(m, 4H), 4.3-4.8(m, 4H), 5.2(d, 1H)5.9(d, 1H),7.1(s, 1H), 7.2-7.4(d, 2H), 8.3-8.5(d, 2H).14. Reacted at room temperature for 5 hrs. Eluant proportion 15:85:1.N.m.r. in solvent B:1.6(s, 6H), 3.2-3.7(m, 2H) 4.0(s, 3H), 4.5-4.9(m, 2H), 5.0-5.4(m, 2H),5.2(d, 1H),5.9(d, 1H), 7.1(s, 1H), 7.2-7.4(m, 2H), 7.9-8.0(m, 2H), 8.2-8.5(m, 2H),8.9-9.0(m, 2H).15. The precursor obtained as in Footnote 4 by reductive amination ofpyridine-4-carboxaldehyde. Eluant proportion 20:80:1. Yield 37%. N.m.r. spectrum insolvent B:1.6(s, 6H), 3.6-3.8(m, 2H), 3.9-4.3(m, 2H), 4.4-5.0(m, 2H), 5.2(d, 1H),6.0(d, 1H),7.1(s, 1H), 8.0-8.3(m, 2H), 8.8-9.2(m, 2H).16. Reacted at room temperature for 1.5 hrs. Eluant porportion 25:75:1.N.m.r. in solventB: 1.55(s, 6H), 3.2(s, 3H), 3.2-4.0(m, 6H), 3.9(s, 3H), 4.6-4.9(m,2H), 5.2(d, 1H), 5.9(d, 1H), 7.1(s, 1H), 7.2(d, 2H), 8.2(d, 2H).17. The precursor was obtained as in Footnote 4 by reductive amination of2-methoxy-acetaldehyde, 2 hrs at room temperature. Eluant proportion 20:80:1. Yield24%. N.m.r. insolvent B: 1.6(s, 6H), 3.3(s, 3H), 3.2-4.0(m, 8H), 5.2(d, 1H), 5.9(d,1H), 7.05(s, 1H).18. Reacted for 2 hrs. Eluant proportion 25:75:1. N.m.r. in solvent B:1.2(t, 3H),1.55(s, 6H), 2.3(s, 3H), 3.2-3.9(m 4H), 4.7(s, 2H), 4.9(m, 2H),5.2-5.5(m, 3H),5.9-6.5(m, 2H), 7.0-7.5(m, 5H), 7.55(d, 2H), 8.4(d, 2H).19. Reaction time 8 hrs. at 45.degree. C. Eluant proportion 20:80:1.N.m.r. in solvent B:1.1-1.4(m, 6H), 1.6(s, 6H), 2.2-2.4(m, 1H), 3.2-3.6(m, 2H), 4.0(s, 3H),4.2-4.8(m, 2H), 5.2(d, 1H), 5.9(d, 1H), 7.1(s, 1H), 7.3(d, 2H), 8.3(d,2H).20. The precursor was obtained as in Footnote 4, by reductive aminationof acetone(4 eq.). Eluant proportion 20:80:1. Yield 50%. N.m.r. in solvent B:1.2(d, 6H), 1.6(s, 6H),3.2-4.0(m, 5H), 5.2(d, 1H), 5.9(d, 1H), 7.05(d, 1H).21. Reacted at 40-45.degree. C. for 4 hrs. Eluant proportion 25:75:1.N.m.r. in solvent B: 1.2(t,3H), 1.55(s, 6H), 3.5(m, 2H), 3.4-3.8(m, 2H), 3.6(m, 2H), 5.2(d, 1H),5.9(d, 1H), 5.9(s, 2H),7.05(s, 1H), 7.1-7.3(m, 1H), 8.5(d, 1H), 9.3(s, 1H).22. Reacted at room temperature for 1.5 hrs, using Boc-aminoethyl CAZAMCAasstarting material. Eluant proportion 35:65:1. The product was deprotectedby solution inTFA at room temperature for 1/2 hr. N.m.r. in solvent B: 155(s, 6H),2.6-4.0(m, 6H),4.0(s, 3H, 4.6-4.9(m, 2H), 5.3(d, 1H), 6.0(d, 1H), 7.0(s, 1H), 7.2(d,1H), 8.2(d, 1H).23. The precursor was obtained by reductive amination of2-(Boc-amino)acetaldehyde atroom temperature for 2 hrs. Eluant proportion 35:65:1. Yield 20%. N.m.r.in solvent B:1.4(s, 9H), 1.55(s, 6H), 2.7-3.4(m, 4H), 3.6-4.2(m, 4H), 5.2(d, 1H),5.9(d, 1H),7.05(s, 1H). The 2-(Boc-amino)propionaldehyde was obtained by oxidationof2-(Boc-amino)ethanol with pyridinium chlorochromate in CH.sub.2 Cl.sub.2containingsodium acetate, at 0.degree. C. for 2 hrs, and purification bychromatography on silica. Yield 68%.24. Reaction time 2 hrs. at room temperature. Eluant proportion 25:75:1.N.m.r. in solvent B:1.55(s, 6H), 3.2-3.6(m, 2H), 3.8-4.0(m, 2H), 4.0(s, 3H), 4.2-4.5(m, 1H),4.8-5.1(m, 1H),4.6-4.8(m, 2H), 5.2(d, 1H), 5.9(d, 1H), 7.05(s, 1H), 7.3(d, 1H), 8.3(d,1H).25. The precursor was obtained by reductive alkylation of an excess of2-fluoroacetaldehyde inCH.sub.2 Cl.sub.2 solution, for 2.5 hrs. at room temperature. Eluantproportion 25:75:1. Yield 23%.N.m.r. in solvent B: 1.55(s, 6H), 3.4-3.6(m, 2H), 3.6-4.1(m, 2H),4.4-4.6(m, 1H),5.0-5.2(m, 1H), 5.2(d, 1H), 5.9(d, 1H), 7.1(s, 1H).26. Reaction carried out for 3 hrs. at room temperature and 1 hr. at40.degree. C. Eluantproportion 25:75:1. N.m.r. in solvent B: 1.2(m, 3H), 1.55(s, 6H),3.5-3.9(m, 4H),3.95(s, 3H), 4.7(s, 2H), 5.25(d, 1H), 5.9(d, 1H), 7.0(s, 1H), 7.3-7.7(m,1H), 8.2(d, 1H),8.65(d, 1H).27. Reaction carried out at 45.degree. C. for 1 hr. Eluant proportion20:80:1.N.m.r. in solvent B: 1.5(s, 6H), 3.1(s, 3H), 3.2(d, 1H), 3.5(d, 1H),3.9(s, 3H), 4.5(d, 1H),4.65(d, 1H), 5.2(d, 1H), 5.85(d, 1H), 7.05(s, 1H), 7.0-7.2(m, 2H),8.2-8.4(m, 2H).28. The CAZAMCA starting material was obtained as follows: A solution of4-chloro-N-methylbutyramide in CH.sub.2 Cl.sub.2 was treated dropwise with silverfluoroborate ina mixture of CH.sub.2 Cl.sub.2 and benzene at -20.degree. C., and thereaction was allowedto warm to room temperature. The silver chloride precipitate was filteredoff, and theproduct was purified by silica chromatography, eluting with CH.sub.2Cl.sub.2 /MeOH, 4:1,to yield N-methyliminobutyrolactone fluoroborate, 71%, which wasconverted to free baseby reaction with 1 eq. of 1,8-diazabicylo[5,4,0]undec-7-ene (DBU) inCH.sub.2 Cl.sub.2(yield 62% of free base). This product was reacted with7-[2-(2-aminothiazol-4-yl)-2-[(Z)-1-carboxy-1-methylethoxyimino]acetamido]-3-iodomethylceph-3-em-4-carboxylicacid in acetonitrile at room temperature for 1/2 hr. to give thecorresponding3-[N-(1,2,3,4-tetrahydo-2-furylidene)-N-methylaminomethyl]cephem, whichwashydrolysed with 4 eq. of sodium bicarbonate in water at room temperaturefor 1/2 hr.,and the product purified by preparative HPLC eluting with MeOH/H.sub.2O/HOAc, 20:80:1 toyield the required starting material, yield 45%.29. Reaction carried out in DMF/H.sub.2 O, 1:1, at 40.degree. C. for 4hours. Eluant proportion25:75:1. N.m.r. in solvent B: 1.1(m, 3H), 1.55(s, 6H), 2.5(s, 3H),3.0-3.7(m, 4H),3.8(s, 3H), 4.6(s, 2H), 5.2(d, 1H), 5.9(d, 1H), 7.0-7.05(m, 3H), 8.2(d,1H).30. Reaction at room temperature for 1.5 hrs. Eluant proportion 25:75:1.N.m.r. insolvent B: 1.6(s, 6H), 3.2-3.6(m, 4H), 4.0(s, 3H), 4.0-4.3(m, 1H),4.5-4.8(m, 2H),5.0-5.2(m, 1H), 5.2(d, 1H), 5.5-6.0(m, 1H), 5.9(d, 1H), 7.1(s, 2H),7.2(d, 2H), 8.3(d, 1H).31. The precursor CAZAMCA derivative was a mixture of N-allyl andN-propyl,obtained by reductive alkylation of acrylaldehyde, as in Footnote 4,using 1.2 eq.of the aldehyde, and reacting at room temperature for 1 hr. Eluantproportion20:80:1. This mixture was used to prepare the final product, purificationof whichseparated the N-allyl and N-propyl analogues.32. Eluant proportion 25:75:1. N.m.r. in solvent B: 0.9(t, 3H), 1.6(s,6H), 3.1-3.8(m, 4H),3.9(s, 3H), 4.5-4.8(m, 2H), 5.2(d, 1H), 5.9(d, 1H), 7.0(s, 1H), 7.2(d,2H), 8.2(d, 2H).33. Reacted overnight at room temperature. Eluant proportion 20:80:1.N.m.r. in solventB: 1.5(s, 6H), 2.6-3.0(m, 2H), 3.95(d, 1H), 3.25(d, 1H), 3.95(s, 3H),4.5-4.8(m, 2H),5.2(d, 1H), 5.9(d, 1H), 7.0(s, 1H), 7.4(d, 2H), 8.4(d, 2H).34. The precursor was obtained as in Footnote 4 by Michael addition ofacrylonitrile toCAZAMCA The product was purified by HPLC, eluant proportion 15:85:1.Yield 31%.N.m.r. in solvent B: 1.55(s, 6H), 2.8-3.1(m, 2H), 3.2-3.4(m, 2H),3.5-3.8(m, 2H), 3.8-4.2(m, 2H), 5.2(d, 1H), 5.9(d, 1H), 7.0(s, 1H).35. This compound obtained by reacting the product of Example 67 withformic aceticanhydride in MeOH at room temperature for 1/2 hr. Eluant proportion20:80:1. N.m.r.in solvent B: 1.55(s, 6H), 3.2-3.8(m, 6H), 3.9(s, 3H), 3.5-3.8(m, 2H),5.2(d, 1H),5.9(d, 1H), 7.0(s, 1H), 7.2(d, 2H), 8.0(s, 1H), 8.2(d, 2H).
EXAMPLES 76-87
The appropriate 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-(substituted oximino) acetamido]ceph- 3-em-4-carboxylic acid and the appropriate amino-substituted quaternary heterocycle were reacted together at 85.degree. C. at pH 7.0-7.5. The pH was maintained with this range by the addition of dilute HOAc or NaHCO.sub.3 solution as necessary. The product was purified by preparative HPLC, using a mixture of MeOH/H.sub.2 O/HOAc as eluant.
The following compounds were prepared:
__________________________________________________________________________ ##STR102##Example R.sup.11 R R.sup.1 Yield Footnotes__________________________________________________________________________76. Me H ##STR103## 11 177. Me H ##STR104## 6 278 C(CH.sub.3).sub.2 COOH H ##STR105## 8 379 Me H ##STR106## 7 4, 580 Me Me ##STR107## 31 6, 781. C(CH.sub.3).sub.2 COOH H ##STR108## 6 882. C(CH.sub.3).sub.2 COOH H ##STR109## 9 983. C(CH.sub.3).sub.2 COOH Me ##STR110## 13 1084. C(CH.sub.3).sub.2 COOH Et ##STR111## 13 11, 1285. C(CH.sub.3).sub.2 COOH (CH.sub.2).sub.2 NH.sub.2 ##STR112## 7 13, 14, 1586. C(CH.sub.3).sub.2 COOH Et ##STR113## 4 16, 1787. C(CH.sub.3).sub.2 COOH H ##STR114## 12 18__________________________________________________________________________Footnotes1. Reaction time 3 hrs. Eluant proportion 15-20: 85-80:1. N.m.r. insolvent B: 3.55(s, 2H), 3.95(s, 3H),4.4(s, 3H), 4.4(s, 2H), 5.2(d, 1H), 5.8(d, 1H), 7.05(s, 1H), 7.25(d, 1H),7.6(d, 1H), 7.85(t, 1H),8.6(d, 1H), 8.8(d, 1H), 9.8(s, 1H).2. Reaction time 2 hrs. Eluant proportions as in Footnote 1. N.m.r. insolvent B; 3.55(s, 2H),4.0(s, 3H), 4.46(s, 3H), 4.46(s, 2H), 5.15(d, 1H), 5.75(d, 1H), 7.0(s,1H), 7.0(d, 1H), 7.45(d, 1H),7.8-8.05(m, 2H), 9.3(d, 1H), 9.5(d, 1H).3. Reaction time 2 hrs. Eluant proportion 30-35: 70-65:1. N.m.r. insolvent B: 1.55(s, 6H), 3.55(s, 2H),4.4(s, 3H), 4.4(s, 2H), 5.15(d, 1H), 5.8(d, 1H), 7.05(s, 1H), 7.15(d,1H), 7.55(d, 1H), 7.75(t, 1H),8.5(d, 1H), 8.7(d, 1H), 9.7(s, 1H).4. Eluant proportion 20-25: 80-75:1. N.m.r. in solvent B: 3.55(s, 2H),3.95(s, 3H), 4.4(s, 2H),5.2(d, 1H), 5.5(s, 2H), 5.8(d, 1H), 7.05(s, 1H), 7.3(d, 1H), 7.65(d, 1H),7.85(t, 1H), 8.6(d, 1H),8.8(d, 1H), 9.8(s, 1H).5. The 5-amino-2-carbamoylmethylisoquinolinium chloride used as startingmaterial was obtained byalkylation of 5-aminoisoquinoline (1 eq.) with 2-chloroacetamide (1 eq.)in DMF overnight at 45.degree. C.The precipitate was washed with DMF, then CH.sub.2 Cl.sub.2, and dried.Yield 76%. N.m.r. in solvent B:5.45(s, 2H), 7.4(dd, 1H), 7.6(d, 1H), 7.75(t, 1H), 8.45(d, 1H), 8.6(d,1H), 9.8(s, 1H).6. Eluant proportion as in Footnote 4. N.m.r. in solvent B: 2.8(s, 3H),3.6(s, 2H), 4.0(s, 3H),4.15(s, 2H), 4.45(s, 3H), 5.15(d, 1H), 5.75(d, 1H), 7.0(s, 1H), 7.6-8.1,8.4-8.6 and9.6-9.9(3m, 6H).7. The 2-methyl-5-methylaminoisoquinolinium iodide used as startingmaterial was obtained byreacting 5-methylaminoisiquinoline with a large excess of methyl iodideat room temperature for 17hrs. in CH.sub.2 Cl.sub.2. The solvents were evaporated to give therequired salt. Yield 100%.N.m.r. in solvent B: 4.0(s, 3H), 4.4(s, 3H), 7.05(d, 1H), 7.55(d, 1H),7.85(t, 1H), 8.6(s, 2H),9.8(s, 1H).8. Reaction time 3 hrs. Eluant proportion 25:75:1. N.m.r. in solvent B:1.5(s, 6H), 3.55(s, 2H),4.45(s, 2H), 5.2(d, 1H), 5.45(s, 2H), 5.85(d, 1H), 7.05(s, 1H),7.15-7.4(m, 1H), 7.5-7.9(m, 2H),8.3-8.9(m, 2H), 9.7-9.9(m, 1H).9. Eluant proportions 20-25-30:80-75-70:1. N.m.r. in solvent B; 1.5(s,6H), 3.6(s, 2H),4.5(s, 5H), 5.2(d, 1H), 5.85(d, 1H), 7.0(s, 1H), 6.7-7.1(m, 1H), 7.5(d,1H), 7.8-8.1(m, 2H),9.3-9.6(m, 2H).10. Eluant proportion 20-30:80-70:1. N.m.r. in solvent B: 1.5(s, 6H),2.8(s, 3H), 3.55(s, 2H),4.4(s, 2H), 4.45(s, 3H), 5.1(d, 1H), 5.8(d, 1H), 7.05(s, 1H), 7.0-7.2,7.6-8.2, 8.4-8.6, and9.6-10.0 (4 .times. m, 6H).11. Eluant proportion 25-30:75-70:1. N.m.r. in solvent B: 0.9(t, 3H),1.55(s, 6H), 3.2(q, 2H),3.55(s, 2H), 4.4(s, 2H), 4.45(s, 3H), 5.05(d, 1H), 5.75(d, 1H), 7.0(s,1H), 7.0-7.2, 7.6-8.2,8.4-8.6 and 9.6-10.0 (4 .times. m, 6H).12. The 5-ethylamino-1-methylisoquinolinium iodide was prepared by theprocess described inFootnote 7. N.m.r. in solvent B: 1.3(t, 3H), 3.3(q, 2H), 4.4(s, 3H),7.1(d, 1H), 7.55(d, 1H),7.8(t, 1H), 8.6(d, 1H), 8.65(d, 1h), 9.75(s, 1H).13. The quaternary heterocycle used as starting material was5-(2-Boc-aminoethylamino)-2-methylisoquinolinium methosulphate. Reaction time 2.5 hrs. Eluantproportion 30-40: 70-60:1.N.m.r. in solvent B: 1.3(s, 9H), 1.55(s, 6H), 3.1-3.5(m, 4H), 3.55(s,2H), 4.4(s, 2H),4.45(s, 3H), 5.05(d, 1H), 5.75(d, 1H), 7.0(s, 1H), 7.0-7.2, 7.6-8.2,8.4-8.6 and9.6-10.0 (4 .times. m, 6H).14. The product was deprotected by dissolving in CH.sub.2 Cl.sub.2 /TFA(1:2) for 10 minutes atroom temperature. N.m.r. in solvent B: 1.55(s, 6H), 2.9-3.5(m, 4H),3.55(s, 2H), 4.4(s, 2H),4.45(s, 3H), 5.05(d, 1H), 5.75(d, 1H), 7.0(s, 1H), 7.0-7.2, 7.6-8.2,8.4-8.6 and9.6-10.0 (4 .times. m, 6H).15. The 5-(2-Boc-aminoethylamino)-2-methylisoquinolinium methosulphatestarting material wasprepared as follows: 5-Hydroxyisoquinoline (1 eg.) and ethylene diamine(10 eq.), were reactedin a Bucherer reaction, in water with sodium hydrogen sulphite (2 eq.) at160.degree. C. for 20 hrs.The product was purified by chromatography on silica, eluting withCH.sub.2 Cl.sub.2 /MeOH,97-90:3-10. Yield 26%. N.m.r. in d.sub.6 -DMSO: 2.9(t, 2H), 3.2(dt, 2H),6.3(t, 1H),6.75(dd, 1H), 7.25(d, 1H), 7.45(t, 1H), 8.05(d, 1H), 8.4(d, 1H), 9.15(s,1H). The Boc derivativewas formed by reacting the above product with 0,0.sup.1 -di-tert-butylcarbonic anhydride(1.5 eq.) in THF for 1 hr. The Boc compound was purified bychromatography on silica, elutingwith ether. Yield 66%. N.m.r. in CDCl.sub.3 : 1.45(s, 9H), 3.2-3.7(m,4H), 4.8-5.1(m, 1H),5.3-5.6(m, 1H), 6.7(dd, 1H), 7.3(d, 1H), 7.45(t, 1H), 7.65(d, 1H),8.45(d, 1H), 9.15(s, 1H).This Boc product was quaternised with dimethyl sulphate, in CH.sub.2Cl.sub.2 at room temperatureor 2 hrs. The solvent was evaporated, and the residue was washed withether. N.m.r. in CDCl.sub.3 ;1.45(s, 9H), 3.2(m, 2H), 3.55(m, 2H), 3.8(s, 6H), 4.5(s, 3H), 6.7(d, 1H),7.3(d, 1H), 7.55(t, 1H),8.1(d, 1H), 8.5(d, 1H), 9.5(s, 1H).16. Eluant proportion as in Footnote 11. N.m.r. in solvent B: 0.95(t,3H), 1.55(s, 6H), 3.25(q, 2H),3.6(s, 2H), 4.4(s, 2H), 4.6(s, 3H), 5.05(d, 1H), 5.8(d, 1H), 7.0(s, 1H),7.2-7.5, 7.8-8.2,9.2-9.5(3 .times. m, 6H).17. The 5-ethylamino-1-methylquinolinium iodide used as starting materialobtained by theprocess described in Footnote 7, starting from 5-ethylaminoquinoline, 72hrs at room temperature.Yield 100%. N.m.r. in solvent B: 1.3(t, 3H), 3.35(q, 2H), 4.5(s, 3H),6.85(d, 1H), 7.35(d, 1H),7.7-8.1(m, 2H), 9.25(d, 1H), 9.45(d, 1H).18. Eluant proportion as in Footnote 10. N.m.r. in solvent B: 1.55(s,6H), 3.55(s, 2H) 4.3(s, 2H),4.5(s, 3H), 5.2(d, 1H), 5.85(d, 1H), 7.0(s, 1H), 7.2(d, 1H), 7.6(dd, 1H),7.8(dd, 1H), 8.15(d, 1H),8.6(d, 1H), 8.85(d, 1H).
EXAMPLES 88-106
The appropriate quaternary chloro-hetercycle (1 eq.) was reacted with 3-aminomethyl 7-[2-(2-aminothiazol-4-yl)-2-[(Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid (1 eq.) in DMF/H.sub.2 O (2:1). The solvents were evaporated and the residue was purified by HPLC, eluting with a mixture of MeOH/H.sub.2 O/HOAc. The following compounds were prepared:
__________________________________________________________________________ ##STR115##Example R Yield Footnotes__________________________________________________________________________ ##STR116## 33 1, 2, 38 ##STR117## 23 3, 4, 3890. ##STR118## 18 5, 6, 38 ##STR119## 26 7, 8, 38 ##STR120## 35 9, 10, 38 ##STR121## 40 11, 12, 38 ##STR122## 28 13, 14, 38 ##STR123## 40 15, 16, 38 ##STR124## 45 17, 18, 38 ##STR125## 44 19, 20, 38 ##STR126## 15 21, 22, 38 ##STR127## 14 23, 24, 38100. ##STR128## 21 25, 26, 38101. ##STR129## 36 27, 28, 38102. ##STR130## 35 29, 30, 38103. ##STR131## 37 31, 32, 38104. ##STR132## 32 33, 34, 38105. ##STR133## 17 35, 36, 38106. ##STR134## 21 37, 38__________________________________________________________________________Footnotes1. Reaction time 3 hrs. at room temperature. Eluant proportion 20:80:1.N.m.r. insolvent B: 1.55(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 4.35(s, 2H), 5.2(d, 1H),5.3(s, 2H),5.85(d, 1H), 7.05(s, 1H), 6.9-7.2 and 8.2-8.6 (2 .times. m, 4H).2. The starting pyridinium salt was obtained by condensation of4-methylthio-pyridine and 4-chloromethyl-2-quanidinothiazole hydrochloride inEtOH/DMF(5:1) under reflux overnight. The product was purified by silicachromatography,eluting with CH.sub.2 Cl.sub.2 /MeOH, 100-70:30. Yield 20%. N.m.r. insolvent B: 2.65(s, 3H), 5.65(s, 2H), 7.55(s, 1H), 7.85 and 8.9 (2 .times.d, 4H).3. Reaction time 1 hr. at room temperature. Eluant proportion as inFootnote 1.N.m.r. in solvent B: 1.55(s, 6H), 3.45(d, 1H), 3.75(d, 1H), 4.4(s, 2H),5.1(s, 2H),5.25(d, 1H), 5.9(d, 1H), 5.3(s, 1H), 7.1(s, 1H), 7.0-7.3(m, 2H),8.1-8.5(m, 2H).4. The starting pyridinium salt was obtained by the process in Footnote2, byalkylating 4-methylsulphonylpyrimidine with 6-chloromethyluracil. Yield47%.N.m.r. in solvent B: 2.7(s, 3H), 5.4(s, 3H), 7.9(d, 2H), 8.7(d, 2H).5. Reaction time 2.5 hrs. at room temperature. Eluant proportion 30:70:1.N.m.r. insolvent B: 1.55(s, 6H), 3.55(s, 2H), 4.4(s, 2H), 5.2(d, 1H), 5.9(d, 1H),6.0(s, 2H),6.9-7.2(m, 2H), 7.1(s, 1H), 7.95(s, 4H), 8.1-8.5(m, 2H).6. The starting pyridinium salt was obtained by the process of Footnote2. Yield 30%.N.m.r. in solvent B: 2.65(s, 3H), 6.25(s, 2H), 7.9(s, 4H), 7.9 and 8.7(2.times. d, 4H).7. Reaction as Footnote 3. N.m.r. in solvent B: 1.6(s, 6H), 3.6(s, 2H),4.4(s, 2H),5.3(d, 1H), 5.4(s, 2H), 5.95(s, 1H), 7.1(s, 1H), 7.0-7.3(m, 2H),8.2-8.6(m, 2H),8.85(s, 1H).8. The starting pyridinium salt was obtained by the process of Footnote2, carriedout at room temperature for 40 hrs. Yield 81%. N.m.r. in solvent B:2.7(s, 3H),5.65(s, 2H), 7.95(d, 2H), 8.8(d, 2H), 8.8(s, 1H).9. Reaction time 2 hrs. Eluant proportion 25:75:1. N.m.r. in solvent B:1.55(s, 6H),3.25(s, 3H), 3.45(s, 3H), 3.55(s, 2H), 4.35(s, 2H), 4.7(s, 4H), 5.25(d,1H), 5.9(d, 1H),7.0(s, 1H), 6.8-7.2(m, 2H), 7.8(s, 1H), 7.9-8.3(m, 2H).10. The starting pyridinium salt was obtained by the process of Footnote2. Yield46%. N.m.r. in solvent B: 2.7(s, 3H), 3.2(s, 3H), 3.45(s, 3H), 4.85(s,4H),7.85(s, 1H), 7.95(d, 2H), 8.5(d, 2H).11. Reaction time 1.5 hrs. at room temperature, using a 50% excess ofthepyridinium salt. Eluant proportion 30:70:1. N.m.r. in solvent B: 1.55(s,6H),2.55(s, 3H), 3.4(d, 1H), 3.7(d, 1H), 4.35(s, 2H), 5.25(d, 1H), 5.9(d,1H),5.45(s, 2H), 7.05(s, 1H), 7.55(s, 1H), 6.9-7.2(m, 2H), 8.2-8.6(m, 2H).12. The starting pyridinium salt was obtained by the process of Footnote2, in EtOHas solvent. Yield 19%. Nmr in solvent B: 2.6(s, 3H), 2.7(s, 3H), 5.75(s,2H), 7.75(s,1H), 7.95(d, 2H), 8.8(d, 2H).13. Reaction time 1.5 hrs. at room temperature. Eluant proportion30:70:1. N.m.r. insolvent B: 1.3(t, 3H), 1.55(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 4.35(s, 2H),4.4(q, 2H),5.25(d, 1H), 5.9(d, 1H), 5.6(s, 2H), 7.15(s, 1H), 6.9-7.2(m, 2H), 8.25(s,1H), 8.2-8.6(m, 2H).14. The starting pyridinium salt was obtained by the process of Footnote2. Yield70%. N.m.r. in solvent B: 1.3(t, 3H), 2.65(s, 3H), 4.35(q, 2H), 5.86(s,2H), 7.9(d,2H), 8.25(s, 1H), 8.8(d, 2H).15. Reaction time 2 hrs. at room temperature. Eluant proportion asFootnote 1.N.m.r. in solvent B: 1.55(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 4.35(s, 2H),5.2(d, 1H),5.5(s, 2H), 5.85(d, 1H), 7.1(s, 1H) 6.9-7.2(m, 2H), 8.05(s, 1H),8.1-8.4(m, 2H).16. The starting pyridinium salt was obtained by the process of Footnote2. Yield18%. N.m.r. in solvent B: 2.75(s, 3H), 5.9(s, 2H), 7.8(d, 2H), 8.25(s,1H), 8.85(d,2H).17. Reaction time 1/2 hr. at room temperature. Eluant proportion asFootnote 5.N.m.r. in solvent B: 1.55(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 4.25(d, 1H),4.5(d, 1H),5.2(d, 1H), 5.6(s, 2H), 5.9(d, 1H), 7.05(s, 1H), 7.0-7.2(m, 2H), 8.4(s,1H), 8.2-8.6(m, 2H).18. The starting pyridinium salt is obtained by the process of Footnote2. Yield 90%.N.m.r. in solvent B: 2.7(s, 3H), 5.9(s, 2H), 8.05(d, 2H), 8.45(s, 1H),8.8(d, 2H).19. Reaction time 2 hrs. at room temperature. Eluant proportion asFootnote 2.N.m.r. in solvent B; 1.55(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 4.25(d, 1H),4.5(d, 1H),5.25(d, 1H), 5.9(d, 1H), 5.95(s, 2H), 7.15(s, 1H), 6.9-7.2(d, 2H),8.3-8.7(d, 2H), 9.4(s,1H).20. The starting pyridinium salt was obtained by the process of Footnote2, but for48 hrs. at room temperature. Yield 48%. N.m.r. in solvent B: 2.55(s, 3H),2.7(s, 3H),6.25(s, 2H), 7.95(d, 2H), 8.95(d, 2H), 9.45(s, 1H).21. Reaction time 3.5 hrs. at room temperature. Eluant proportion20-30:80-70:1.N.m.r. in solvent B: 1.55(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 4.4(s, 2H),5.25(d, 1H), 5.4(s,1H), 5.9(d, 1H), 7.15(s, 1H), 6.9-7.2(m, 2H), 7.35(s, 1H), 8.55(s, 1H),8.2-8.6(m,2H).22. The starting pyridinium salt was obtained by the process of Footnote2. Yield39%. N.m.r. in solvent B: 2.7(s, 3H), 5.7(s, 2H), 7.4(s, 1H), 7.95(d,2H), 8.5(s, 1H),8.8(d, 2H).23. The 3-aminomethyl cephem was alkylated with the corresponding2-(Boc-amino)thiazole derivative, at room temperature for 40 minutes. The Bocprotectinggroup was removed by dissolving the product in TFA/CH.sub.2 Cl.sub.2(4:1) at roomtemperature for 50 minutes. Eluant proportion 30-35:70-65:1. N.m.r. insolvent B:1.55(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 4.4(s, 2H), 5.2(s, 2H), 5.25(d, 1H),5.9(d, 1H),7.05(s, 1H), 6.9-7.2(m, 2H), 8.0-8.4(m, 2H).24. The starting Boc-amino-chloromethylthiazole was obtained by theprocess ofFootnote 2. Yield 65%. N.m.r. in solvent B; 1.45(s, 9H), 2.7(s, 3H),5.7(s, 2H),7.35(s, 1H), 7.9(d, 2H), 8.75(d, 2H).25. Reaction time 1.25 hrs. at room temperature. Eluant proportion30-40:70-60:1.N.m.r. in solvent B: 1.5(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 4.25(d, 1h),4.5(d, 1H), 5.15(d,1H), 5.85(d, 1H), 6.0(s, 2H), 7.1(s, 1H), 7.0-7.2(m, 2H), 7.5-7.7(m, 2H),7.7-7.9(m,2H), 8.2-8.6(m, 2H).26. The starting pyridinium salt was obtained by the process of Footnote2, at roomtemperature overnight followed by 3 hrs. under reflux in EtOH/DMF (3:2).Thesolvent was evaporated, and the product was precipitated with ether.Yield 54%.N.m.r. in solvent B: 2.7(s, 2H), 6.35(s, 2H), 7.55(m, 2H), 7.75(m, 2H),8.0(d, 2H),8.9(d, 2H).27. Reaction time 40 minutes at room temperature. Eluant proportion asFootnote 1.N.m.r. in solvent B: 1.55(s, 6H), 2.4(dt, 2H), 3.4(d, 1H), 3.7(d, 1H),4.4(m, 4H),5.2(d, 1H), 5.5(t, 1H), 5.9(d, 1H), 6.9(d, 1H), 7.1(s, 1H), 7.0-7.2(m,2H), 7.15(t,1H), 7.55(dt, 1H), 7.9(dd, 1H), 8.2-8.6(m, 2H).28. The starting pyridinium salt was obtained by the process of Footnote2, at roomtemperature overnight followed by 24 hrs. under reflux. Yield 20%. N.m.r.in sol-vent B: 2.5(dt, 2H), 2.7(s, 3H), 4.7(t, 2H), 5.5(t, 1H), 6.9(d, 1H),7.15(t, 1H), 7.55(dt,1H), 7.9(dd, 1H), 8.0(d, 2H), 8.9(d, 2H).29. Reaction time 2 hrs. at room temperature. Eluant proportion10-25:90-75:1.N.m.r. in solvent B: 1.55(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 3.9(s, 3H),4.4(s, 2H), 5.2(d,1H), 5.85(d, 1H), 5.85(s, 2H), 7.0(s, 1H), 7.7(d, 1H), 7.75(d, 1h),7.0-7.4(m, 2H), 8.2-8.6(m, 2H).30. The starting pyridinium salt was obtained by the process of Footnote2, at roomtemperature for 96 hrs. Yield 30%. N.m.r. in solvent B: 2.7(s, 3H),3.95(s, 3H),6.2(s, 2H), 7.7(d, 1H), 7.75(d, 1H), 7.95(d, 1H), 8.85(d, 1H).31. Reaction time 3 hrs. room temperature. Eluant proportion10-20:90-80:1. N.m.r.in solvent B: 1.55(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 4.4(s, 2H), 5.2(d,1H), 5.75(s, 2H),5.85(d, 1H), 7.05(s, 1H), 7.65(s, 2H), 7.0-7.3(m, 2H), 8.2-8.4(m, 2H).32. The starting pyridinium salt was obtained by the process of Footnote2. Yield83%. N.m.r. in solvent B: 2.75(s, 3H), 6.15(s, 2H), 7.75(s, 2H), 8.05(d,2H), 8.85(d,2H).33. Reaction time 1.5 hrs. at room temperature. Eluant proportion asFootnote 1.N.m.r. in solvent B: 1.5(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 4.4(s, 2H),5.15(d, 1H), 5.55(s,2H), 5.85(d, 1H), 7.05(s, 1H), 7.05(dd, 1H), 7.0-7.2(m, 2H), 7.4(d, 1H),7.55(d, 1H),8.2-8.6(m, 2H).34. The starting pyridinium salt was obtained by the process of Footnote2, at roomtemperature overnight followed by 4 hrs. under reflux. Yield 46%. N.m.r.in sol-vent B: 2.65(s, 3H), 5.9(s, 2H), 7.05(dd, 1H), 7.4(d, 1H), 7.55(d, 1H),7.85(d, 2H),8.8(d, 2H).35. Reaction time 3 hrs. at room temperature. Eluant proportion40-50:60-50:1.N.m.r. in solvent B: 1.55(s, 6H), 3.4(d, 1H), 3.7(d, 1H), 4.3(d, 1H),4.5(d, 1H), 5.2(d,1H), 5.85(s, 2H), 5.9(d, 1H), 7.05(s, 1H), 7.0-7.3(m, 2H), 7.7(d, 1H),8.1(dd, 1H),8.5(d, 1H), 8.2-8.6(m, 2H).36. The starting pyridinium salt was obtained by the process of Footnote2, heatingunder reflux for 6 hrs., then overnight at room temperature. Yield 77%.N.m.r. insolvent B: 2.7(s, 3H), 6.1(s, 2H), 7.7(d, 1H), 7.95(d, 2H), 8.1(dd, 1H),8.5(d, 1H),8.85(d, 2H).37. This compound obtained by reduction of the product of Example 105withtitanium trichloride (5 eq.) in MeOH/TFA (99:1) for 1 hr. at roomtemperature.Eluant proportion as Footnote 1. N.m.r. in solvent B: 1.55(s, 6H), 3.4(d,1H), 3.7(d,1H), 4.3(d, 1H), 4.5(d, 1H), 5.2(d, 1H), 5.9(d, 1H), 5.9(s, 2H), 7.1(s,1H), 7.0-7.3(m,2H), 7.44(d, 1H), 7.85(dd, 1H), 7.9(d, 1H) 8.2-8.6(m, 2H).38. The quaternised thiomethyl heterocycle salt was oxidised with MCPBAinCH.sub.2 Cl.sub.2 containing a catalytic amount of TFA at 0.degree. C. toroom temperature for upto 4 hours, to give the corresponding sulphinyl compound, often alsocontaining sul-phone and sulphide, but not further purified.
EXAMPLE 107
An activated ester was prepared from (2-aminothiazol-4-yl)-(2-methoxyimino)acetic acid (1.1 eq.) by reaction with hydroxybenzotriazole (1.1 eq.) and N,N'-dicyclohexylcarbodi-imide (1.1 eq.) in DMF, at room temperature for 2 hrs. To this ester was added a solution of 7-amino-3-[N-ethyl-N-(l-methyl-4-pyridinio)aminomethyl]ceph-3-em-4-carboxylate (1.0 eq.) in DMF and Et3N (2 eq.). The reaction mixture was stirred at room temperature for 2 hrs. and filtered, and the solvents were evaporated. The product, 7-[2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetamido]-3-(N-ethyl)-[N-(1-methylpyridinio)]aminomethylceph-3-em-4-carboxylate, was purified by preparative HPLC eluting with MeOH/H.sub.2 O/HOAc, 10:90:1. N.m.r. in solvent B: 1.0-1.3 (m, 3H), 3.0-3.8 (m, 4H), 3.9 (s, 3H), 3.95 (s, 3H), 4.5-4.7 (m, 2H), 5.2 (d, 1H), 5.8 (d, 1H), 7.0 (s, 1H), 7.15 (d, 2H), 8.2 (d, 2H).
The cephem starting material was obtained as follows:
3-Aminomethyl-7-(Boc-amino)ceph-3-em-4-carboxylic acid (1 eq.) was dissolved in MEOH, with Et.sub.3 N (1 eq.) and NaBH.sub.3 CN (1 eq.) at room temperature. Acetaldehyde (1 eq.) was added over 15 minutes at room temperature, the solvent was evaporated, and the product, 7-(Boc-amino)-3-ethylaminomethylceph-3-em-4-carboxylic acid, was purified by preparative HPLC, eluting with MEOH/H.sub.2 O/HOAC, 30:70:1. Yield 59%, contaminated with some disubstituted material.
The above product (1 eq.) was treated DMF/H.sub.2 O (3:2) with 4-chloro-1-methylpyridinium iodide (1 eq.) in presence of NAHCO.sub.3 (3 eq.). After 2 hrs. at room temperature, the solvents were evaporated and the product was purified by preparative HPLC, eluting with MeOH/H.sub.2 O/HOAc, 30:70:1. Yield 20%. N.m.r. in d.sub.6 -DMSO: 1.0-1.2 (m, 3H), 1.4 (s, 9H), 2.8-3.8 (m, 4H), 3.9 (s, 3H), 4.2-4.8 (m, 2H), 4.9 (d, 1H), 5.2 (dd, 1H), 7.7 (d, 2H), 8.2 (d, 2H).
The above product was deprotected by dissolving in TFA and leaving for 1/2 hr. at room temperature. The TFA was evaporated, the residue triturated with ether to give the required starting material. Yield 84%. N.m.r. in solvent B: 0.9-1.3 (m, 3H), 3.2-3.8 (m, 4H), 3.9 (s, 3H), 4.5-4.8 (m, 2H), 5.2 (s, 2H), 7.15 (d, 2H), 8.25 (d, 2H).
EXAMPLES 108-115
Method A
The appropriate quaternised heterocyclic starting material (0.28mM) dissolved in a mixture of CH.sub.3 CN/H.sub.2 O (1:1; 2 ml) was added to a stirred solution of 3-ethylaminomethyl-7-[2-(2-aminothiazol-4-yl)-(Z)-2-(1-carboxycyclobut-1-yloxyimino)acetamido]ceph-3-em-4-carboxylic acid (0.28 mm) in CH.sub.3 CN/H.sub.2 O (1:1; 3 ml) at room temperature under an atmosphere of argon. Sodium bicarbonate (1.14 mM) was added to the reaction mixture which was stirred for 5 hr. and then poured into water (100 ml). The resulting solution was adjusted to pH 3.5 with acetic acid and then concentrated by rotary evaporation under vacuum at 20.degree. C. The resulting aqueous solution was filtered and then diluted with water (100 ml) before loading on to an HP20SS column (20mm.times.350mm). The product was eluted from the resin by increasing amounts of CH.sub.3 CN in H.sub.2 O. Fractions containing the required product were combined and concentrated in vacuo and the aqueous residues freeze dried to give lyophilized solids.
Method B
The appropriate quaternised heterocyclic starting material (0.29 mM) in dimethylformamide (2 ml) was added at room temperature to a stirred solution of the N-ethylaminocephem-carboxylic acid of Method A (0.28 mM) in DMF (3 ml.). Triethylamine (0.56mm) was then added to the mixture and the reaction was allowed to continue for 16 hr. The mixture was then poured into water (100 ml) and the pH adjusted to 3.5 with acetic acid. After filtering, the solution was applied to an HP20SS column (20 mm.times.350 mm) and the product was eluted with increasing amounts of CH.sub.3 CN in H.sub.2 O. The fractions containing the required product were pooled and concentrated by rotary evaporation at 20.degree. C. to give an aqueous concentrate which was then freeze dried.
Using these general processes the following compounds were prepared:
__________________________________________________________________________ ##STR135##Example R Yield % Method Footnotes__________________________________________________________________________108 ##STR136## 14 A 1, 2109 ##STR137## 31 A 3, 4110 ##STR138## 42 B 5, 6111 ##STR139## 44 B 7, 8112 ##STR140## 37 A 9, 10113 ##STR141## 16 B 11, 12__________________________________________________________________________
The same general processes were also utilized with the corresponding aminomethyl intermediate to synthesize the following unalkylated 3-aminomethyl derivatives:
______________________________________ Foot-Example R Yield % Method notes______________________________________114 ##STR142## 19 B 13. 14115 ##STR143## 25 A 15, 16______________________________________Footnotes1. The starting material was 6-chloro-1-methyl-1,2,4-triazolo[4,3-b]pyridazinium iodide.2. n.m.r. in solvent A: 9.62(s, 1H); 8.48(d, 1H); 8.15(m, 1H); 6.73(s,1H); 5.70(d, 1H); 5.03(d, 1H); 4.6(m, 2H); 4.14(s, 3H); 3.5-3.8(m,2H); 3.15-3.5(ABq, 2H); 2.15-2.80(m, 6H); 1.05-1.25(m, 3H).3. The starting material was 3-chloro-1-methyl-pyridaziniumiodide4. n.m.r. in solvent A: 8.95(m, 1H); 8.05(m, 2H); 6.72(s, 1H);5.75(d, 1H); 5.08(d, 1H); 4.45-4.80(ABq, 2H); 4.30(s, 1H); 3.56-3.75(m, 2H); 3.20-3.55(ABq, 2H); 2.20-2.60(m, 6H); 1.05-1.30(m,3H).5. The starting material was 1-methyl-4-methylthiothieno[2,3-d]pyrimidine tetrafluoroborate.6. n.m.r. in solvent A: 8.86(s, 1H); 7.93(d, 1H); 7.80(d, 1H); 6.74(s,1H); 5.79(d, 1H); 5.11(d, 1H); 4.74-5.39(ABq, 2H); 4.02(m, 5H);3.15-3.59(ABq, 2H); 2.20-2.48(m, 6H); 1.20-1.42(m, 3H).7. The starting material was 2,3-dihydro-7-methylthiothiazolo[2,3-a]pyrimidinium tetrafluoroborate.8. n.m.r. in solvent A: 8.3(m, 1H); 6.90-7.4(m, 1H); 6.74(s, 1H);5.75(m, 1H); 5.05(m, 1H); 4.65-5.0(ABq, 2H); 4.50-4.65(m, 2H);3.75-3.95(m, 2H); 3.55-3.75(t, 2H); 3.0-3.55(ABq, 2H), 2.20-2.5(m, 6H); 1.1(t, 3H).9. Prepared using 4-chloro-1-(trans-3-chloroalkyl)pyridiniumtosylate as starting material (see Example 151, Footnote 38) andmethod A, except that:DMF was used instead of CH.sub.3 CN and the reaction was workedup by adjusting the pH to 7 with HOAc, concentrating to drynessunder vacuum at 20.degree. and redissolving in water prior to HP20SSchromatography.10. n.m.r. in solvent A: 8.22(d, 2H); 7.51(m, 1H); 7.10(m, 1H);6.75(s, 1H); 6.73(d, 1H); 6.25(m, 1H); 5.73(d, 1H); 5.07(d, 1H);4.81(d, 2H); 4.73, 4.49(ABq, 2H); 3.65(m, 2H); 3.45, 3.13(ABq,2H); 2.36(m, 4H); 1.86(m, 2H)); 1.12(t, 3H).11. The starting material was 1-(4-dimethylaminosulphonylbenzyl)-4-methanesulphinyl pyridinium tetrafluoroborate, which wasobtained as follows:4-Dimethylsulphamoylbenzyl bromide (0.005M) was mixed with4-methylthiopyridine (0.005M) and reacted exothermically, andwas then left to stand overnight. The product was dissolved inCH.sub.2 Cl.sub.2, and after standing for 3 hrs. was added dropwise torapidly stirred ether to precipitate 1-(4-dimethylsulphamoyl-benzyl)-4-methylthiopyridinium bromide, which was converted tothe corresponding tetrafluoroborate salt with silver tetra-fluoroborate.The pyridinium tetrafluoroborate (1.34 mM) was dissolved inCH.sub.2 Cl.sub.2 (15 ml) and treated at 0.degree. with sufficient TFA toobtaina solution, then MCPBA (1.34 mM) was added. The mixture wasstirred at 0.degree. for 15 minutes then at room temperature for 4 hrs.,and the product was precipitated by the dropwise addition of etherto give 1-(4-dimethylsulphamoyl)benzyl-4-methylsulphinyl-pyridinium tetrafluoroborate, the required starting material.12. n.m.r. in solvent A: 8.31(m, 2H); 7.3-7.90(m, 6H); 7.08(m,1H); 6.73(s, 1H), 5.76(d, 1H); 5.48(s, 2H); 5.06(d, 1H); 4.35-4.85(m, 2H), 3.40-3.80(m, 2H); 3.10-3.44(ABq, 2H); 2.56(s, 6H);2.10-2.50(m, 6H); 1.0-1.25(m, 3H).13. See Footnote 1.14. n.m.r. in solvent A: 9.65(s, 1H); 8.35(d, 1H); 7.51(d, 1H);6.76(s, 1H); 5.80(d, 1H); 5.08(d, 1H); 4.05-4.50(ABq, 2H); 4.17(s,3H); 3.36-3.76(ABq, 2H), 2.20-2.47(m, 6H)15. See Footnote 5.16. n.m.r. in solvent A: 8.85(s, 1H); 7.90(m, 1H); 6.75(s, 1H);5.81(d, 1H); 5.08(d, 1H); 4.46-4.95(ABq, 2H); 4.04(s, 3H);3.30-3.70(ABq, 2H), 2.17-2.46(m, 6H).
The 3-ethylaminomethyl-7-[2-(2-aminothiazol-4-yl)-(Z)-2-(1-carboxycyclobut-1-yloxyimino)acetamido]ceph-3-em-4-carboxylic acid used as starting material was prepared as follows:
3-Aminomethyl-7-[2-(2-aminothiazol-4-yl)-(Z)-2-(1-carboxycyclobut-1-yloxyimino)acetamido]ceph-3-em-4-carboxylic acid (10 g; 2.016.times.10.sup.-2 mole) was stirred in MeOH (1.0 l.) and to this mixture was added triethylamine (2.8 ml; 2.016.times.10.sup.-2 mole) dropwise, which resulted in a clear solution. Sodium cyanoborohydride (1.266 g; 2.016.times.10.sup.-2 mole) was added in one portion to the above solution followed by 8g of activated molecular sieves (4A). A solution of acetaldehyde (1.125 ml in 75 ml methanol) was added to the reaction mixture, under an argon atmosphere, over a period of 40 minutes using an infusion pump and the status of the ensuing reaction was monitored by HPLC (Spherisorb 5ODS, 4 mm.times.30 cm; MeOH/H.sub.2 O/TFA (40:60:0.2 by volume), 2 ml/min flow rate and monitoring by u.v. at 270 nm). Reaction was continued for 1 hr., the mixture was then filtered, and the filtrate was concentrated to dryness on a rotary evaporator under high vacuum. The residue was dissolved in water (100 ml) and loaded onto an HP20 macroreticular resin (3 l.) previously equilibrated with 0.2% TFA in water. The column was washed with 0.2% TFA in water (2 l.) and then water (5 l.) followed by increasing amounts of CH.sub.3 CN in water. Fractions (2 l.) were collected and the desired product eluted with 15% CH.sub.3 CN/H.sub.2 O. Fractions (21-23) containing the required product were pooled, and concentrated on a rotary evaporator under high vacuum to give a clear solution (100 ml) which was freeze dried to yield 3 g lyophilised solid. Further fractions containing less pure amounts of the required product were also combined and concentrated to give a crude sample (2 g) of the N-ethyl derivative. This crude sample was purified by medium pressure chromatography on HP20SS resin (350 mm.times.50 mm; flow rate 49 ml/min, 250 ml fractions) to give an additional pure sample (1 g) of the required intermediate.
n.m.r. in solvent A: 6.75 (s, 1H); 5.75 (d, 1H); 5.05 (d, 1H), 3.25-3.85 (m, 4H=two AB quartets); 2.95 (q, 2H), 2.30-2.50 (m, 6H), 1.15 (t, 3H).
EXAMPLES 116-151
To a solution of 3-aminomethyl-7-C2-(2-aminothiazol-4-yl)-2-C(Z)-1-carboxy-1-methylethoxyamino)acetamido]ceph-3-em-4-carboxylic acid (0.4 mmole) in DMF (11 ml) and water (3 ml) at 0.degree. was added sodium bicarbonate (4.8 mmole) dissolved in the minimum volume of water followed by the appropriate 4-chloropyridinium toluene-p-sulphonate salt (0.4 mmole). After 1 hour the mixture was treated with HOAC (4.8 mmole) and evaporated to dryness under reduced pressure. The residue was dissolved in water and purified by one of the following methods:
(a) HPLD on an octadecylsilane column using MeOH/water/HOAc 40:60:1 by volume as eluant.
(b) Medium pressure liquid chromatography on a column of Diaion HP20SS resin (supplied commercially by Mitsubishi Chemical Industries Ltd.) using increasing concentrations of acetonitrile (from 0% to 30% v/v) in water as eluant.
In each case the eluant containing the required product was evaporated on a Buchi Rotavapor to remove the organic solvent and the resulting aqueous soution was dried by lyophilisation. Using this general method the following compounds were obtained:
______________________________________ ##STR144##Ex-am- Yield Foot-ple R % notes______________________________________116 ##STR145## 70 1117 ##STR146## 37 2118 ##STR147## 36 3119 ##STR148## 18 4120 ##STR149## 36 5121 ##STR150## 43 6122 ##STR151## 62 7123 ##STR152## 52 8124 ##STR153## 25 9125 ##STR154## 17 10126 ##STR155## 20 11127 ##STR156## 32 12128 ##STR157## 19 13129 ##STR158## 13 14130 ##STR159## 30 15131 ##STR160## 6 16132 ##STR161## 35 17133 ##STR162## 44 18134 ##STR163## 6 19135 ##STR164## 22 20136 ##STR165## 11 21137 ##STR166## 22 22138 ##STR167## 47 23, 24139 ##STR168## 23 25, 26140 ##STR169## 30 27141 ##STR170## 25 28142 ##STR171## 15 29143 ##STR172## 9 30144 ##STR173## 27 31145 ##STR174## 14 32146 ##STR175## 17 33147 ##STR176## 20 34148 ##STR177## 37 35149 ##STR178## 18 36150 ##STR179## 5 37151 ##STR180## 20 38______________________________________Footnotes1. n.m.r. in solvent A: 1.40(s, 6H); 3.43(d, 2H); 3.64(d, 1H); 4.30(d,1H); 4.48(d, 1H); 5.15(d, 1H); 5.84(d, 1H); 6.74(s, 1H); 7.06(dd,1H); 7.26(dd, 1H); 7.53(d, 1H); 7.66(q, 1H); 7.89(m, 1H); 8.34(dd,1H); 8.54(dd, 1H).2. n.m.r. in solvent A: 1.46(s, 6H); 3.45(d, 1H); 3.66(d, 1H); 4.35(d,1H); 4.51(d, 1H); 5.17(d, 1H); 5.86(d, 1H); 6.74(s, 1H); 7.10(d, 1H);7.28(d, 1H); 7.86(d, 2H); 8.12(d, 2H); 8.46(d, 1H); 8.64(d, 1H).3. n.m.r. in solvent A: 1.44(s, 6H); 3.45(d, 1H); 3.65(d, 1H); 4.31(d,1H); 4.52(d, 1H); 5.15(d, 1H); 5.85(d, 1H); 6.75(s, 1H); 7.13(dd,1H); 7.31(dd, 1H); 7.68-7.84(m, 2H); 7.90(dd, 1H); 8.07(dd, 1H);8.33(dd, 1H); 8.54(dd, 1H).4. n.m.r. in solvent A: 1.40(s, 6H); 3.34(d, 1H); 3.57(d, 1H); 4.31(d,1H); 4.56(d, 1H); 5.05(d, 1H); 5.72(d, 1H); 6.75(s, 1H); 7.09(dd,1H); 7.53(dd, 1H); 8.20(dd, 1H); 8.34(d, 1H); 8.42(dd, 1H); 8.52(d,1H); 8.60(dd, 1H).5. n.m.r. in solvent A: 1.35(s, 6H); 3.30(d, 1H); 3.57(d, 1H); 4.24(d,1H); 4.44(d, 1H); 5.05(d, 1H); 5.70(d, 1H); 6.65(s, 1H); 7.00(d, 1H);7.15(d, 1H); 7.80(d, 2H); 8.30(d, 2H); 8.30(d, 1H); 8.48(d, 1H).6. A solution of 7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]-3-[1-(4-nitrophenyl)-4-pyridinio-aminomethyl]ceph-3-em-4-carboxylate (110 mg) in a mixture ofwater (12 ml) and DMF (6 ml) was stirred for 1 hr. with 10%palladium on carbon in an atmosphere of hydrogen. The solutionwas filtered through delite, and the filtrate was evaporated todryness. The resulting solid was washed with a mixture of waterand methanol (3:1 v/v, 3 ml), collected by filtration and dried invacuo to yield 3-[1-(4-aminophenyl)-4-pyridinio]aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)-acetamido]ceph-3-em-4-carboxylate (45 mg) having n.m.r. insolvent A: 1.45(s, 6H); 3.35(d, 1H); 3.60(d, 1H); 4.20(d, 1H);4.40(d, 1H); 5.10(d, 1H); 5.78(d, 1H); 6.70(s, 1H); 6.80-8.50(complex, 8H).7. n.m.r. in solvent A: 1.40(s, 6H); 3.35(d, 1H); 3.65(d, 1H);4.27(d, 1H); 4.50(d, 1H); 5.10(d, 1H); 5.78(d, 1H); 6.70(s, 1H);7.00-7.30(m, 2H); 7.70-8.10(m, 2H); 8.20-8.60(m, 4H).8. The method described in Footnote 6 was repeated using 7-(2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)-acetamido]-3-[1-(3-nitrophenyl)-4-pyridinio]aminomethylceph-3-em-4-carboxylate to yield 3-[1-(3-aminophenyl)-4-pyridinio]-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyiminoacetamido]ceph-3-em-4-carboxylate havingn.m.r. in solvent A: 1.44(s, 6H); 3.45(d, 1H); 3.65(d, 1H); 4.34(d,1H); 4.47(d, 1H); 5.15(d, 1H); 5.84(d, 1H); 6.75(s, 1H); 6.90-7.45(m,6H); 8.35(d, 1H); 8.54(d, 1H).9. The toluene-p-sulphonate salt of 7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]-3-[1-(4-tert-butoxycarbonylphenyl)-4-pyridinio]aminomethylceph-3-em-4-carboxylate (320 mg) was dissolved with stirring in TFA (4 ml).After 15 minutes the solution was evaporated to dryness and theresidue was purified by HPLC to yield the toluene-p-sulphonatesalt of 7-[2-(2-aminothiazol-4-yl)-2-[(Z)-1-carboxy-1-methyl-ethoxyimino)acetamido]-3-[1-(4-carboxyphenyl-4-pyridinio]amino-methylceph-3-em-4-carboxylate (73 mg) having n.m.r. in solventA: 1.43(s, 6H); 2.27(s, 3H); 3.43(d, 1H); 3.64(d, 1H); 4.34(d, 1H);4.53(d, 1H); 5.15(d, 1H); 5.82(d, 1H); 6.75(s, 1H); 7.12(d, 3H);7.35(d, 1H); 7.50(d, 2H); 7.77(d, 2H); 8.16(d, 2H); 8.47(d, 1H);8.65(d, 1H).The tert-butyl ester used as starting material in theabove process was obtained by the general method using 1-(4-tert-butoxycarbonylphenyl)-4-chloropyridinium toluene-p-sulphonateas starting material.10. Ice cold TFA (5 ml) was added to a stirred mixture of anisole(1 ml) and 7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methyl-ethoxyimino)acetamido]-3-[1-(4-tert-butoxyphenyl)-4-pyridinio]-aminomethylceph-3-em-4-carboxylate (200 mg). The reactionmixture was allowed to warm to 10.degree. C. and after 90 mins wasevaporated to dryness. Purification of the residue by HPLCyielded 7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methyl-ethoxyimino)acetamido]-3-[1-(4-hydroxyphenyl)-4-pyridinio]-aminomethylceph-3-em-4-carboxylate having n.m.r. in solvent A:1.50(s, 6H); 3.28(d, 1H); 3.60(d, 1H); 4.26(d, 1H); 4.60(d, 1H);5.05(d, 1H); 5.74(d, 1H); 6.75(s, 1H); 6.93(d, 2H); 7.45(d, 2H);6.9-7.5(m, 3H); 8.30(d, 1H); 8.45(d, 1H).The tert-butyl ether used as starting material in the above processwas obtained by the general method using 1-(4-tert-butoxyphenyl)-4-chloropyridinium toluene-p-sulphonate as starting material.11. n.m.r. in solvent A: 1.40(s, 6H); 3.45(d, 1H); 3.65(d, 1H);4.35(d, 1H); 4.50(d, 1H); 5.15(d, 1H); 5.75(d, 1H); 6.75(s, 1H);7.10(dd, 1H); 7.30(m, 1H); 7.90(d, 1H); 8.15(m, 1H); 8.60(d, 1H);8.80(dd, 1H); 8.95(dd, 1H).12. n.m.r. in solvent A: 1.40(s, 6H); 3.35(d, 1H); 3.62(d, 1H);4.25(d, 1H); 4.46(d, 1H); 5.05(d, 1H); 5.80(d, 1H); 6.75(s, 1H);7.04(d, 1H); 7.10(d, 1H); 7.55(dd, 1H); 8.05(d, 1H); 8.35(d, 1H);8.52(d, 1H); 8.65(d, 1H); 8.80(d, 1H).13. n.m.r. in solvent A: 1.40(s, 6H); 3.34(d, 1H); 3.67(d, 1H),4.36(d, 1H); 4.57(d, 1H); 5.07(d, 1H); 5.75(d, 1H); 6.74(s, 1H);7.12(d, 1H); 7.55(d, 1H); 8.15(d, 1H); 8.60(dd, 1H); 8.82(d, 1H);8.95(d, 1H); 9.05(d, 1H).14. n.m.r. in solvent A: 1.42(s, 6H); 3.34(d, 1H); 3.56(d, 1H);4.37(d, 1H); 4.56(d, 1H); 5.07(d, 1H); 5.74(d, 1H); 6.74(s, 1H);7.12(dd, 1H); 7.50(dd, 1H); 7.60(t, 1H); 8.98(d, 2H); 9.14(dd, 1H);9.24(dd, 1H).15. n.m.r. in solvent A: 1.45(s, 6H); 3.35(d, 1H); 3.58(d, 1H);4.35(d, 1H); 4.60(d, 1H); 5.07(d, 1H); 5.74(d, 1H); 6.74(s, 1H);7.14(dd, 1H); 7.58(dd, 1H); 8.26(d, 1H); 8.38(d, 1H); 8.76(dd, 1H);8.86(dd, 1H).16. n.m.r. in solvent A: 1.44(s, 6H); 3.30(d, 1H); 3.56(d, 1H);4.30(d, 1H); 4.60(d, 1H); 5.07(d, 1H); 5.72(d, 1H); 6.72(s, 1H);7.19(dd, 1H); 7.60(dd, 1H); 7.9-8.6(m, 2H); 8.86(m, 2H);9.37(q, 1H).17. n.m.r. in solvent A: 1.45(s, 6H); 3.29(d, 1H); 3.54(d, 1H);4.28(d, 1H); 4.56(d, 1H); 5.05(d, 1H); 5.71(d, 1H); 6.70(dd, 1H);6.75(s, 1H); 6.88(dd, 1H); 7.06(dd, 1H); 7.58(dd, 1H); 7.76(dd, 1H);8.44(dd, 1H); 8.60(dd, 1H).18. n.m.r. in solvent A: 1.44(s, 6H); 3.44(d, 1H); 3.64(d, 1H);4.32(d, 1H); 4.47(d, 1H); 5.16(d, 1H); 5.84(d, 1H); 6.74(s, 1H);7.10(dd, 1H); 7.05(dd, 1H); 7.25(dd, 1H); 7.85(dd, 1H); 8.36(dd,1H); 8.41(dd, 1H); 8.58(dd, 1H).19. n.m.r. in solvent A: 1.43(s, 6H); 3.40(d, 1H); 3.60(d, 1H);4.30(d, 1H); 4.58(d, 1H); 5.13(d, 1H); 5.81(d, 1H) (d, 1H); 6.74(s,1H); 7.00-7.15(m, 2H); 7.20(dd, 1H); 7.46(dd, 1H); 7.54(dd, 1H);8.32(dd, 1H); 8.50(dd, 1H).20. n.m.r. in solvent A: 1.45(s, 6H); 3.46(d, 1H); 3.67(d, 1H);4.34(d, 1H); 4.52(d, 1H); 5.17(d, 1H); 5.85(d, 1H); 6.75(s, 1H);7.08(dd, 1H); 7.24(dd, 1H); 7.50(dd, 1H); 7.80(dd, 1H); 7.98(dd,1H); 8.46(dd, 1H); 8.64(dd, 1H).21. n.m.r. in solvent A: 1.45(s, 6H); 2.10(s, 3H); 3.30(d, 1H);3.55(d, 1H); 4.35(d, 1H); 4.55(d, 1H); 5.05(d, 1H), 5.70(d, 1H);6.75(s, 1H); 7.05(d, 2H); 7.15(d, 2H); 7.95(s, 1H); 8.60(dd, 2H);8.70(dd, 2H).22. n.m.r. in solvent A: 1.43(s, 6H); 3.33(d, 1H); 3.56(d, 1H),4.35(d, 1H); 4.56(d, 1H); 5.06(d, 1H); 5.72(d, 1H), 6.75(s, 1H);7.08(dd, 1H); 7.51(dd, 1H); 7.74(s, 2H); 8.66(dd, 1H); 8.75(dd, 1H).23. Prepared by the general method using 1-(2-benzoxazolyl)-4-methylsulphinylpyridinium chloride in place of the 4-chloropy-ridinium toluene-p-sulphonate.The pyridinium salt was prepared as follows.A solution of 2-chlorobenzoxazole (1.38 g) and 4-methylthio-pyridine (1.13 g) in anhydrous methylene chloride was kept in thedark overnight. The resulting crystalline precipitate of 1-(2-benzoxazolyl)-4-methylthiopyridinium chloride (1.86 g) wascollected by filtration, washed with anhydrous methylene chlorideand dried in vacuo.A solution of 3-chloroperbenzoic acid (127 mg, 85% pure) inmethylene chloride (3 ml) was added dropwise to a stirred icecold solution of 1-(2-benzoxazolyl)-4-methylthiopyridiniumchloride (139 mg) in methylene chloride (3 ml) and TFA (0.10ml). The solution was allowed to warm to 20.degree. C. After 40minutes, toluene (2 ml) was added and the mixture wasevaporated to dryness to yield a mixture of 1-(2-benzoxazolyl)-4-methylsulphinylpyridinium chloride and 3-chlorobenzoic acidwhich was used directly in the general method.24. n.m.r. in solvent A: 1.45(s, 6H); 3.30(d, 1H); 3.65(d, 1H);4.35(d, 1H); 4.62(d, 1H); 5.05(d, 1H); 5.70(d, 1H); 6.74(s, 1H);7.00-7.90(m, 6H); 8.75(dd, 1H); 8.85(dd, 1H).25. Prepared by the process described in Footnote 23, using2-chlorobenzothiazole as starting material in place of 2-chloro-benzoxazole.26. n.m.r. in solvent A: 1.40(s, 6H); 3.33(d, 1H); 3.56(d, 1H);4.37(d, 1H); 4.64(d, 1H); 5.06(d, 1H); 5.72(d, 1H); 6.74(s, 1H);7.15(d, 1H); 7.45-7.70(m, 3H); 8.00(d, 1H); 8.18(d, 1H); 8.80(d,1H); 8.86(d, 1H).27. n.m.r. in solvent A: 1.45(s, 6H); 3.30(d, 1H); 3.55(d, 1H);4.20(d, 1H); 4.45(d, 1H); 5.05(d, 1H); 5.15(d, 1H); 5.25(d, 1H);5.70(d, 1H); 6.75(s, 1H); 6.80-7.20(m, 3H); 8.15(d, 1H); 8.30(d, 1H).28. n.m.r. in solvent A: 1.45(s. 6H), 3.40(d, 1H); 3.52(d, 1H);3.82(s, 3H); 4.28(d, 1H); 4.44(d, 1H); 5.04(d, 1H); 5.75(d, 1H);6.75(s, 1H); 7.00-7.22(m, 3H); 7.35-7.62(m, 3H); 8.30(d, 1H);8.45(d, 1H).29. n.m.r. in solvent A: 1.45(s, 6H); 2.10(s, 3H); 3.30(d, 1H);3.65(d, 1H); 4.29(d, 1H); 4.59(d, 1H); 5.07(d, 1H); 5.71(d, 1H);6.75(s, 1H); 7.07(d, 1H); 7.57(d, 2H); 7.57(d, 1H); 7.80(d, 2H);8.35(d, 1H); 8.50(d, 1H).30. n.m.r. in solvent A: 1.35(s, 6H); 1.45(t, 3H); 3.48(d, 1H);3.66(d, 1H); 4.30(m, 3H); 4.50(d, 1H); 5.17(d, 1H); 5.85(d, 1H);6.75(s, 1H); 7.11(d, 1H); 7.36(d, 1H); 7.82(d, 2H); 9,15(d, 2H);9.50(d, 1H); 9.70(d, 1H).31. n.m.r. in solvent A: 1.40(s, 6H); 2.80(s, 6H); 3.30(d, 1H);3.58(d, 1H); 4.25(d, 1H); 4.50(d, 1H); 5.05(d, 1H); 5.70(d, 1H);6.72(s, 1H); 7.00(d, 1H); 7.30(d, 1H); 7.60(m, 4H); 8.30(d, 1H);8.45(d, 1H).32. n.m.r. in solvent A: 1.45(s, 6H); 3.30(s, 3H); 3.45(d, 1H);3.65(d, 1H); 4.35(d, 1H); 4.50(d, 1H); 5.15(d, 1H); 5.85(d, 1H);6.75(s, 1H); 7.14(d, 1H); 7.33(d, 1H); 7.94(d, 2H); 8.16(d, 2H);8.46(d, 1H); 8.66(d, 1H).33. n.m.r. spectrum in solvent A: 1.45(s, 6H), 3.35(d, 1H); 3.60(d,1H); 4.35(d, 1H); 4.55(d, 1H); 5.05(d, 1H); 5.70(d, 1H), 6.70(s, 1H);7.10(dd, 1H); 7.46(dd, 1H); 8.12(d, 1H); 8.50(d, 1H); 8.80(d, 1H);8.94(d, 1H); 8.97(broad, 1H).34. n.m.r. spectrum in solvent A: 1.45(s, 6H); 2.70(s, 3H); 3.30(d,1H); 3.55(d, 1H); 4.30(d, 1H); 4.60(d, 1H); 5.05(d, 1H); 5.70(d, 1H);6.75(s, 1H); 7.10(d, 1H); 7.60(d, 1H); 7.80(d, 2H); 8.15(d, 2H);8.35(d, 1H), 8.55(d, 1H).35. n.m.r. in solvent A: 1.40(s, 6H); 2.08(m, 2H); 2.50(t, 2H);3.26(d, 1H); 3.50(d, 1H); 4.15(t, 2H); 4.20(d, 1H); 4.42(d, 1H);5.02(d, 1H); 5.69(d, 1H); 6.73(d, 1H); 6.94(d, 1H); 7.35(d, 1H);9.10(d, 1H); 8.25(d, 1H).36. n.m.r. in solvent A: 1.46(s, 6H); 3.26(d, 1H); 3.50(d, 1H);4.20(d, 1H); 4.44(d, 1H); 4.80(d, 2H); 5.05(d, 1H); 5.20(d, 1H);5.32(d, 1H); 5.72(d, 1H); 5.90(d t, 1H); 6.3-6.6(m, 2H); 6.74(s,1H); 6.94(d, 1H); 7.38(d, 1H); 8.16(d, 1H); 8.25(d, 1H).37. n.m.r. in solvent A: 1.40(s, 6H); 3.40(d, 1H); 3.60(d, 1H);4.25(d, 1H); 4.40(d, 1H); 4.90(d, 2H); 5.15(d, 1H); 5.80(d, 1H);6.20(m, 1H); 6.65(d, 1H); 6.75(d, 1H); 6.90(d, 1H); 7.10(d, 1H);8.05(d, 1H); 8.20(d, 1H).38. n.m.r. in solvent A: 1.40(s, 6H); 3.40(d, 1H); 3.60(d, 1H);4.25(d, 1H); 4.35(d, 1H); 4.80(d, 2H); 5.15(d, 1H); 5.85(d, 1H);6.20(m, 1H); 6.20(m, 1H); 6.70(d, 1H); 6.75(s, 1H); 6.90(d, 1H);7.10(d, 1H); 8.10(d, 1H); 8.25(d, 1H).
The 1-substituted-4-chloropyridinium toluene-p-sulphonate salts used as starting materials in the above processes may be prepared as follows:
A solution of the appropriate 1-substituted-4-pyridone (2.5 mmole) and toluene-p-sulphonyl chloride (3.8 mmole) in DMF (10 ml) was stirred at 100.degree. C. for 10 mins. The reaction mixture was cooled and evaporated to dryness. Trituration of the residue with ether yielded the required 4-chloropyridinium toluene-p-sulphonate salt as a solid which was used without further purification in the preparation of the final product. Using the general process the following compounds were prepared:
______________________________________ ##STR181## Starting MaterialR Footnotes for Example Number______________________________________3,4-difluorophenyl 1 1164-cyanophenyl 2 1172-cyanophenyl 3 1183,4-dicyanophenyl 3 1194-nitrophenyl 3 120, 1213-nitrophenyl 4 122, 1234-tert-butoxycarbonylphenyl 5 1244-tert-butoxyphenyl 6 1252-pyridyl 7 1263-pyridyl 4 1275-cyano-2-pyridyl 7 1282-pyrimidinyl 7 1296-chloro-3-pyridazinyl 7 1303-pyridazinyl 8 1312-furyl 9 1323-furyl 10 1332-thienyl 11 1343-thienyl 11 1354-methyl-2-oxazolyl 12 1362-thiazolyl 13 137vinyl 1404-methoxyphenyl 4 1414-acetamidophenyl 14 1424-ethoxycarbonylphenyl 5 1434-dimethylaminocarboxy- 15 144phenyl4-methylsulphonylphenyl 16 1455-trifluoromethyl-2-pyridyl 7 1464-acetoxyphenyl 5 1473-cyanopropyl 17 148penta-2,4-dienyl 18 149cis-3-chlorallyl 19 150trans-3-chloroallyl 19 151______________________________________Footnotes1. The starting material was prepared as follows:A mixture of alpha-pyrone (880 mg.), 3,4-difluoroaniline(1.54 g.) and concentrated hydrochloric acid (0.8 ml.)in water (10 ml.) was stirred under reflux for 2 hours.The reaction mixture was cooled and basified withaqueous ammonia solution. The precipitated product wasfiltered off, washed with cold ethyl acetate (10 ml.)and dried in vacuo to yield 1-(3,4-difluorophenyl)-4-pyridone (1.10 g.) as a buff solid.2. The starting material was prepared as follows:Sodium hydride (410 mg. of a 1% dispersion in mineraloil) was placed in an argon atmosphere, washed free ofoil with -n-pentane and suspended in DMF (10 ml.). Solid4-hydroxypyridine (1.00 g.) was added in small portionsto the suspension. Stirring was continued for 15minutes after the evolution hydrogen gas had ceased andthen a solution of 4-fluorobenzonitrile (1.27 g.) in DMF(3 ml.) was added. The resulting mixture was stirred at110.degree. C. for 30 minutes and then poured whilst still hotinto cold water (60 ml.). After cooling for 30 minutesin ice the crystalline precipitate of 1-(4-cyanophenyl)-4-pyridone (1.45 g.) was collected by filtration, washedwith water, dried in vacuo and recrystallised fromEtOAc.3. The pyridone starting material was prepared bythe method described in Footnote 2, using theappropriate substituted chlorobenzene in place of4-fluorobenzonitrile.4. The starting material was prepared by themethod described in Example 2 using the appropriateaniline in place of 4-fluoroaniline.5. The pyridone starting material was prepared bythe method described in Footnote 2, using theappropriate substituted fluorobenzene in place of4-fluorobenzonitrile.6. The pyridone starting material was prepared asfollows:A mixture of alpha-pyridone (440 mg.), 4-tert-butoxy-aniline (755 mg.) and HOAc (0.262 ml.) was stirred at100.degree. C. for 10 minutes in an argon atmosphere. Thecooled reaction mixture was evaporated to dryness, usingthe azeotropic evaporation of added toluene to removethe last traces of HOAc. The residue was purified bychromatography on a column of Kieselgel 60, eluting withmethylene chloride, then 95:5 v/v methylene chloride;methanol, to give 1-(4-tert-butoxyphenyl)-4-pyridone(340 mg.) having n.m.r. in CDC1.sub.3 :1.40(s, 9H), 6.50(d, 2H), 7.0-7.3(m, 4H), 7.60(d, 2H).7. The pyridone starting material was preparedby the method described in Footnote 2, using theappropriate 2-chloroheterocycle in place of 4-fluoro-benzonitrile.8. The pyridone starting material was prepared asfollows:A solution of 1-(6-chloro-3-pyridazinyl)-4-pyridone(1.00 g.) and sodium acetate (0.50 g.) in EtOH (50 ml.)and water (3 ml.) containing 10% palladium on carbon(200 mg.) was stirred for 75 minutes in an atmosphereof hydrogen.The mixture was filtered through kieselguhrand the filtrate was evaporated to dryness.Recrystallisation of the residue from EtOAc yielded1-(3-pyridazinyl)-4-pyridone, (350 mg.) having n.m.r. insolvent A:6.35(d, 2H), 7.95(q, 1H), 8.20(q, 1H), 8.50(d, 2H),9.30(q, 1H).9. The pyridone starting material was prepared asfollows:Sodium hydride (1.69 g. of a 1% dispersion in mineraloil) was placed in an argon atmosphere, washed free ofoil with -n-pentane and suspended in DMSO (27 ml.).Solid 4-hydroxypyridine (4.12 g.) was added in smallportions to the stirred suspension. Stirring wascontinued for 15 minutes after the evolution of hydrogengas had ceased. Powdered methyl 5-nitro-2-furoate(6.18 g.) was then added and stirring was continued for48 hours. The reaction mixture was evaporated todryness and the dark brown residue was extracted with95:5 v/v methylene chloride/MeOH (3 .times. 100 ml.). Theextracts were passed through a column of Kieselgel 60(50 mg.) and the eluent was evaporated to dryness. Theresidue was dissolved in methylene chloride (100 ml.)and the solution was filtered. Evaporation of thefiltrate to a volume of ca 20 ml. deposited pale browncrystals of the methyl ester, 1-(5-methoxycarbonyl-2-furyl)-4-pyridone (3.07 g.), having m.p. 194 - 6.degree. C.Chromatography of the mother liquor on Kieselgel 60afforded a further crop (863 mg.) of the same product.The methyl ester (3.95 g.) was stirred inmethanol (78 ml.) in an argon atmosphere, while asolution of potassium hydroxide (4.87 g.) in water(39 ml.) was added. The methyl ester rapidly dissolved.Stirring was continued overnight and the reactionmixture was evaporated to dryness. The residue wasdissolved in the minimum volume of water and acidifiedwith N-hydrochloric acid to pH. 1. The resultingprecipitate was isolated by centrifugation, washed withwater then acetone, and dried in vacuo to yield theacid, 1-(5-carboxy-2-furyl)-4-pyridone (3.74 g.)A mixture of the acid (3.74 g.), copper bronze(1.64 g.) and quinoline (8 ml.) was stirred at 210.degree. C. inan argon atmosphere until the evolution of carbondioxide ceased (ca. 30 minutes). The cooled reactionmixture was dissolved in EtOAc and filtered throughkieselguhr. The filtrate was evaporated to dryness andthe residue was purified by chromatography on Kieselgel60 (30 g.) eluting initially with methylene chloride toremove quinoline then with 95:5 v/v methylenechloride/MeOH to afford 1-(2-furyl)-4-pyridone (2.20 g.).10. The pyridone starting material was prepared asfollows:A mixture of 3-bromofuran (2.50 g.), freshly sublimed4-hydroxypyridine (1.61 g.), copper powder (2.20 g.) andpowdered anhydrous potassium carbonate (4.80 g.) in DMF(25 ml.) was stirred at 150.degree. C. for 2 hours in an argonatmosphere. The cooled reaction mixture was filteredthrough kieselguhr and the filtrate was evaporated todryness. The residue was extracted with warm 95:5 v/vchloroform/MeOH (100 ml.). The resulting solution wasapplied to a column of Kieselgel 60 (30 g.). Elutionwith the same solvent gave 1-(3-furyl)-4-pyridone(1.17 g.).11. The pyridone starting material was prepared bythe method described in footnote 10, using theappropriate bromothiophene in place of 3-bromofuran.12. The pyridone starting material was prepared bythe method described in footnote 2, using 4-methyl-2-methylsulphinyloxazole in place of 4-fluorobenzo-nitrile.13. The pyridone starting material was prepared bythe method described in footnote 2, using 2-bromo-thiazole in place of 4-fluorobenzonitrile.14. The starting material was prepared as follows:A solution of 1-(4-nitrophenyl)-4-pyridone (500 mg) inEtOH (5 ml) and DMF (5 ml) was stirred at 80.degree. C. for 30minutes with 10% palladium on carbon (150 mg) in anatmosphere of hydrogen. The reaction mixture wascooled and filtered through kieselguhr. The filtrate wasevaporated to dryness and the solid residue was stirredovernight with a mixture of acetic anhydride (1 ml),pyridine (3 ml) and DMF (3 ml). The reaction mixture wasevaporated to dryness to yield 1-(4-acetamidophenyl)-4-pyridone (420 mg.) as a white solid.15. The pyridone starting material was prepared asfollows:A mixture of 1-(4-ethoxycarbonylphenyl)-4-pyridone(1.0 g) and 33% w/v dimethylamine in ethanol was stirredduring the dropwise addition of a solution of sodiumethoxide [prepared by the addition of sodium metal(50 mg) to ethanol (2.5 ml)]. Stirring was continuedovernight, then the reaction mixture was evaporated todryness to yield 1-(4-dimethylcarbamoylphenyl)-4-pyridone which was used without purification.16. The pyridone starting material was prepared asfollows:Using the method described in Example 2,4-methyl-thioaniline was converted into 1-(4-methylthiophenyl)-4-pyridone. A mixture of the pyridone (1.80 mg), HOAc(14 ml) and 100 volumes aqueous hydrogen peroxide (4.5 ml)were stirred at 70.degree. C. for 2 hours. The cooled reactionmixture was diluted with water and extracted with ethylacetate. The ethyl acetate solution was washed withaqueous sodium metabisulphite, dried with sodiumsulphate, and evaporated to yield 1-(4-methylsulphonyl-phenyl)-4-pyridone (1.15 g) which was used withoutfurther purification.17. The pyridone starting material was prepared bythe method described in footnote 2, using 4-bromobutyro-nitrile in place of 4-fluorobenzonitrile.18. The pyridone starting material was prepared asfollows:A mixture of 2-chloropenta-2,4-diene (1.49 g), 4-hydroxy-pyridine (1.39 g), powdered anhydrous potassium carbonate(4.14 g) and DMF (10 ml) was stirred for 90 minutes at80.degree. C. in an argon atmosphere. The reaction mixture wascooled and the DMF was evaporated. The residue wasextracted with 9:1 v.v methylene chloride/methanol(200 ml). The extacts were filtered through a pad ofKieselgel 60 (30 g) and the filtrates were evaporated todryness to give a brown oil. Chromatography of thisoil on Kieselgel 60, eluting initially with methylenechloride, then 19:1 v/v methylene chloride/methanol,afforded 1-(penta-2,4-dienyl)-4-pyridone (1.36 g) as apale orange oil.19. The pyridone starting materials were preparedas follows:A solution of 1,3-dichloropropene (cis/trans mixture,1.97 ml) and 4-hydroxypyridine (2.00 g) in acetone (25 ml)was stirred with powdered anhydrous potassium carbonate(2.90 g) for 1 hour under reflux. Further portions of1,3-dichloropropene (1.97 ml) and potassium carbonate(2.90 g) were added and the reaction mixture was stirredfor a further hour under reflux. The cooled reactionmixture was filtered and the solid residue was washedwell with acetone. The combined acetone filtrates wereevaporated to dryness. The oily residue waschromatographed on a column of Kieselgel 60 (100 g)eluting with methylene chloride, then 19:1 v/v methylenechloride/methanol, to separate the cis and trans isomersof the product, 2-(3-chlorallyl)-4-pyridone.
EXAMPLES 152-158
To a solution of 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-[(Z)-1-carboxy-1-methylethoxyamino)acetamido]ceph-3-em-4-carboxylic acid (0.4 mmole) in DMF (11 ml.) and water (3 ml) at 0.degree. was added sodium bicarbonate (4.8 mmole) dissolved in the minimum volume of water, followed by a solution of the appropriate pyridinium salt (0.5 mmole) in DMF (5 ml). After 1 hour the mixture was treated with HOAc (4.8 mmole) and evaporated to dryness under reduced pressure. The residue was dissolved in water and purified by chromatography on Diaion HP20SS resin.
Using this general process the following compounds were prepared
__________________________________________________________________________ ##STR182##Example R Yield % Footnotes__________________________________________________________________________152 ##STR183## 10 1153 ##STR184## 16 2154 ##STR185## 30 3155 ##STR186## 24 4156 ##STR187## 44 5157 ##STR188## 15 6158 ##STR189## 8 7__________________________________________________________________________1. n.m.r. in solvent A: 1.40(s, 6H); 3.30(d, 1H); 3.50(d, 1H); 4.20(d,1H); 4.40(d, 1H);4.95(d, 2H); 5.05(d, 1H); 5.70(d, 1H); 6.70(s, 1H); 6.95(d, 1H); 7.25(d,1H); 7.65(d,2H); 8.10(d, 1H); 8.15(d, 2H); 8.30(d, 1H).2. n.m.r. in solvent A: 1.45(s, 6H); 3.27(d 1H); 3.50(d, 1H); 4.21(d,1H); 4.43(d, 1H);4.95(d, 2H); 5.04(d, 1H); 5.68(d, 1H); 6.28(m, 1H); 6.74(s, 1H);6.90-7.45(m, 3H);8.02(d, 1H); 8.20(d, 1H).3. n.m.r. in solvent A: 1.40(m, 12H); 3.24(d, 1H); 3.45(m, 5H); 4.20(d,1H); 4.48(d,1H); 4.90 (broad, 2H); 5.05(d, 1H); 5.70(d, 1H); 6.70(s, 1H); 6.80(m,1H); 6.95(m,2H); 7.50(dd, 1H); 8.10(d, 1H); 8.28(d, 1H).4. n.m.r. in solvent A: 1.44(s, 6H); 3.55(m 10H); 4.25(d, 1H); 4.40(d,1H); 4.90(d,2H); 5.15(d, 1H); 5.83(d, 1H); 6.65(m, 2H); 6.75(s, 1H); 6.95(d, 1H);7.07(d, 1H);8.10(d, 1H); 8.27(d, 1H).5. n.m.r. in solvent A: 0.85(t, 3H), 1.35(m, 2H); 1.40(s, 6H); 3.05(m,2H); 3.25(d,1H); 3.50(d, 1H); 4.20(d, 1H); 4.45(d, 1H); 4.90(d, 1H); 5.05(d, 1H);5.70(d, 1H);5.85(d, 1H); 6.75(s, 1H); 6.80(m, 1H); 6.95(dd, 1H); 7.40(dd, 1H);8.10(d, 1H); 8.20(d, 1H).6. n.m.r. in solvent A: 1.40(s, 6H); 3.25(d, 1H); 3.50(d, 1H); 4.20(d,1H); 4.50(d, 1H);4.95(d, 2H); 5.05,(d, 1H); 5.70(d, 1H); 5.80(d, 1H); 6.75(s, 1H); 6.80(m,1H); 7.00(dd, 1H); 7.45(dd, 1H); 8.10(d, 1H); 8.25(d, 1H).7. n.m.r. in solvent A: 1.45(s, 6H); 1.95(s, 3H); 3.40(d, 1H); 3.60(d,1H); 4.25(d, 1H);4.40(d, 1H); 4.90(s, 2H); 5.15(d, 1H); 5.30(s, 1H); 5.85(d, 1H); 6.75(s,1H); 6.95(d,1H); 7.10(d, 1H); 8.00(d, 1H); 8.20(d, 1H).
The pyridinium salts used as starting materials in the above process may be prepared as follows:
A solution of 3-chloroperbenzoic acid (1.05 mmole) in anhydrous methylene chloride (2 ml) was added dropwise to a stirred solution at the appropriate 1-substituted-4-methylthiopyridinium salt (0.5 mmole) in methylene chloride (5 ml) and TFA (0.1 ml) at 0.degree.. The reaction mixture was allowed to warm to room temperature over 1 hour and was then evaporated to dryness. The residue was washed with ether to remove 3-chlorobenzoic acid and the insoluble residue, a mixture of the appropriate 4-methylsulphinyl- and 4-methylsulphonylpyridinium salts, was used directly in the above process.
Using this general procedure the following compounds were prepared:
______________________________________ ##STR190## Starting MaterialR X Footnotes for Example No.______________________________________4-nitrocinnamyl Cl 1 152trans-3-cyanoallyl BF.sub.4 2 153trans-3-(2-piperidino- Br 3 154carboxy)allyltrans-3-(1-morpholino- Br 4 155carboxy)allyltrans-3-(N-propyl- Br 4 156aminocarboxy)allyltrans-3-(amino- Br 4 157carboxy)allyl .sub.-- E-3-cyano-2-methyl- Br 4 158allyl______________________________________Footnotes1. The 1-(4-nitrocinnamyl)-4-methylthiopyridiniumchloride starting material was prepared as follows. Asolution of 4-nitrocinnamyl chloride (181 mg) and4-methylthiopyridine (125 mg) in acetonitrile (1 ml) waskept in the dark for 48 hours. The solvent wasevaporated and the residue was triturated with ether togive the required starting material as an off-whitesolid (170 mg).2. The 1-(trans-3-cyanoallyl)-4-methylthio-pyridinium tetrafluoroborate starting material wasprepared as follows. A solution of 4-bromocrotono-nitrile (660 mg) and 4-methylthiopyridine (565 mg) inanhydrous methylene chloride (3 ml) was kept in the darkfor 48 hours. The resulting black crystalline solid(888 mg) was filtered off and dried in vacuo. Thissolid was dissolved in a mixture of water (20 ml) and DMF(10 ml). To this stirred solution was added a solution ofsilver tetrafluoroborate (636 mg) in water (3 ml). After5 minutes the solution was filtered through celite andevaporated to dryness. The resulting gum was dissolvedin water (20 ml), refiltered and freeze-dried to yieldthe starting material as a fluffy orange solid (810 mg).3. The 1-[trans-3(1-piperidinocarboxy)]allyl-4-methylthiopyridinium bromide starting material wasprepared as follows. A solution of trans-3-(1-piperidinocarboxy)allyl bromide (440 mg) and 4-methyl-thiopyridine (240 mg) in acetonitrile (5 ml) was kept inthe dark for 24 hours. The solvent was evaporated andthe residue was washed with ether to afford the requiredstarting material as a gum.4. The required starting material was prepared,using the procedure in Footnote 3, by reacting 4-methyl-thiopyridine with the appropriate substituted allylbromide.
EXAMPLES 159-160
Using the general processes described in Example 116, with the appropriate 3-aminomethylcephalosporin and pyridinium salt starting materials the following compounds were prepared.
______________________________________ ##STR191## YieldExample R % Footnotes______________________________________159 ##STR192## 17 1160 ##STR193## 15 2______________________________________Footnotes1. n.m.r. in solvent A: 2.40(m, 6H); 3.40(d,1H); 3.60(d, 1H); 4.25(d, 1H); 4.38(d, 1H); 4.80(d,1H); 5.15(d, 1H); 5.85(d, 1H); 6.20(m, 1H); 6.70(d,1H); 6.75(s, 1H); 6.90(d, 1H); 7.10(d, 1H); 8.05(d,1H); 8.25(d, 1H).2. n.m.r. in solvent A: 1.96(s, 3H); 2.40(m,6H); 3.42(d, 1H); 3.65(d, 1H); 4.27(d, 1H); 4.40(d,1H); 4.95(s, 2H); 5.16(d, 1H); 5.32(s, 1H); 5.85(d,1H); 6.74(s, 1H); 6.97(d, 1H); 7.10(d, 1H); 8.04(d,1H); 8.25(d, 1H).
EXAMPLES 161-171
The general process described in Example 1 was repeated using the appropriate heterocyclic starting material. The reactions were carried out in DMF/water mixtures in the presence of NaHCO.sub.3 at the temperature in the range ambient to 75.degree. for 0.4-4 hours. The product was purified on a octadecylsilane column and the following compounds were thus prepared.
__________________________________________________________________________ ##STR194## YieldExample R % Footnotes__________________________________________________________________________161 CH.sub.3 7 1, 2, 3162 NHCH.sub.2 CHCH.sub.2 18 4, 5, 6163 NHCH(CH.sub.3).sub.2 22 7, 8, 9154 N(CH.sub.3).sub.2 25 10, 11, 12165 ##STR195## 14 13, 14, 15166 ##STR196## 4.5 16, 17, 18167 NHCH.sub.2 CH.sub. 2 NH.sub.2 12 19, 20, 21, 22168 NHCH.sub.2 CH.sub.2 OH 11 23, 24, 25169 NHCH.sub.2 CH.sub.2 OCH.sub.3 6 26, 27, 28170 SCH.sub.3 32 29, 30, 31171 SCH.sub.2 CH.sub.2 NHCOCH.sub.3 20 32, 33, 34__________________________________________________________________________Footnotes1. The starting material was prepared by reaction of 3-chloro-6-methylpyridazine with trimethloxonium tetrafluoroborate in CH.sub.2 Cl.sub.2 15hours at ambienttemperature to give 3-chloro-1,6-dimethylpyridazinium tetrafluoroborate.2. HPLC eluant MeOH/water/HOAc 15:84:1, v/v/v.3. n.m.r. in solvent B; 1.53(br.s, 6H); 2.62(s, 3H); 3.6(m, 2H); 4.14(s,3H); 5.15(d,1H); 4.85(d, 1H); 7.05(s, 1H); 7.55(d, 1H); 7.85(d, 1H).4. (a) The starting material was obtained as follows. Reaction of 3,6-difluoropyridazine with trimethyloxonium tetrafluoroborate gave 3,6-difluoro-1-methylpyridazinium tetrafluoroborate.(b) To this compound (1.5 mmole) in acetonitrile (5 ml) was added asolution of allylamine (1.5 mmole) in acetonitrile (3 ml) andNaHCO.sub.3. After 2 hours at ambient temperature the mixture wasfiltered, and the filtrate was evaporated to yield an oil, whichcrystallised to give 3-fluoro-1-methyl-6-(allylamino)-pyridaziniumtetrafluoroborate.5. HPLC eluant MeOH/water/HOAc 25:74:1 v/v/v.6. n.m.r. in solvent B: 1.56(br.s, 6H); 3.55(m, 2H); 3.78(s, 3H); 4.0(m,2H); 3.95(d, 1H); 4.5(d, 1H); 5.15(m, 3H); 5.83(m, 2H); 7.06(s, 1H);7.42(s, 2H).7. The starting material was prepared by reaction of 3,6-difluoro-1-methylpyridazinium tetrafluoroborate (Footnote 4a) with 2 equivalents ofisopropylamine in acetonitrile at ambient temperature, to give 3-fluoro-1-methyl-6-isopropylamino-pyridazinium tetrafluoroborate.8. HPLC eluant MeOH/water/HOAc 30:69:1 v/v/v.9. n.m.r. in solvent B: 1.21(s, 3H); 1.28(s, 3H); 1.54(br.s, 6H); 3.5(m,2H); 3.76(s,3H); 3.85(d, 1H); 4.0(m, 1H); 4.45(d, 1H); 5.15(d, 1H); 5.83(d, 1H);7.05(s,1H); 7.4(d, 1H); 7.67(d, 1H).10. The starting material was prepared as follows. To a stirred solutionof3,6-difluoro-1-methyl-pyridazinium tetrafluoroborate (2.00 mmoles,Footnote4a) in acetonitrile under argon at room temperature was added MgO (2.0mmoles) followed by a solution of dimethylamine in toluene (4.0 mmoles).After 2 hours, the mixture was filtered, and the filtrate was evaporatedtoan oil, which crystallised after trituration with ether, to give3-fluoro-1-methyl-6-(N,N-dimethylamino)pyridazinium tetrafluoroborate.11. HPLC eluant MeOH/water/HOAc 22.5:77.5:1 v/v/v12. n.m.r. in solvent B: 1.53(br.s, 6H); 3.06(s, 6H); 3.45(d, 1H); 3.7(d,1H); 3.89(s, 3H); 4.05(d, 1H); 4.5(d, 1H); 5.13(d, 1H); 5.80(d, 1H); 7.05(s, 1H);7.32(d,1H); 7.52(d, 1H).13. The starting material was prepared by the method described inFootnote 7,using p-nitrobenzyl-amine, to give 3-fluoro-1-methyl-6-(p-nitrobenzylamino)-pyridazinium tetrafluoroborate.14. HPLC eluant MeOH/water/HOAc 35:64:1 v/v/v.15. n.m.r. in solvent B: 1.53(br.s, 6H); 3.5(m, 2H); 3.86(s, 3H); 3.95(d,1H);4.45(d, 1H); 5.14(d, 1H); 5.8(d, 1H); 7.05(s, 1H); 7.36(br.s, 2H);7.62(d, 2H);8.18(d, 2H).16. The starting material was prepared by the method described inFootnote 4 togive 3-fluoro-1-methyl-6-(2-thenylamino)pyridazinium tetrafluoroborate.17. HPLC eluant MeOH/aqueous ammonium carbonate (2 g/l) 30:70 v/v.18. n.m.r. in solvent B: 1.53(s, 6H); 3.55(m, 2H); 3.93(s, 3H); 3.95(d,1H); 4.45(d,1H); 4.95(s, 2H); 5.15(d, 1H); 5.8(d, 1H); 7.05(s, 1H); 6.97-7.7(m, 5H).19. The starting material was prepared by the method described inFootnote 4using N-Boc ethylene diamine.20. HPLC eluant MeOH/aqueous ammonium carbonate (2 g/l), 30:70 v/v.21. Cleavage of the Boc protecting group was achieved after the HPLC bytreatment with TFA for 5 minutes.22. n.m.r. in solvent B: 1.56(s, 6H); 3.1(m, 2H); 3.85(m, 4H); 3.94(s,3H); 4.64(m,2H); 5.27(d, 1H); 5.98(d, 1H); 7.06(s, 1H); 8.05(d, 1H); 8.85(d, 1H).23. The starting material was prepared by a slow addition of a solutionofethanolamine in acetontrile under argon, followed by NaHCO.sub.3. After1 hour at ambient temperature the mixture was filtered, and the filtratewasevaporated to an oil, which crystallised to give 3-fluoro-6-(2-hydroxyethyl-amino)-1-methyl pyridazinium tetrafluoroborate.24. HPLC eluant MeOH/water/HOAc 20:79:1 v/v/v.25. n.m.r. in solvent B: 1.53(s, 6H); 3.54(m, 6H); 3.73(s, 3H); 3.95(d,1H); 4.4(d,1H); 5.1(d, 1H); 5.8(d, 1H); 7.06(s, 1H); 7.35(d, 1H); 7.55(d, 1H).26. The starting material, 3-fluoro-6-(2-methoxy-ethylamino)-1-methyl-pyridazinium tetrafluorobate was prepared by the method described in Foot-note 7 using 2-methoxyethylamine.27. HPLC eluant MeOH/water/HOAc 25:74:1 v/v/v.28. n.m.r. in solvent B: 1.53(s, 6H); 3.23(s, 3H); 3.5(m, 6H); 3.73(s,3H); 3.95(d,1H); 4.5(d, 1H); 5.15(d, 1H); 3.85(d, 1H); 7.07(s, 1H); 7.4(d, 1H);7.7(d, 1H).29. The starting material was obtained by reaction of 3,6-difluoro-1-methylpyri-dazinium tetrafluoroborate (Footnote 4a) with sodium thiomethylate inacetonitrile for 1 hour at ambient temperature to give 3-fluoro-1-methyl-6methylthiopyridazinium tetrafluoroborate.30. The crude condensation product was purified by chromatography onDiaionHP20 resin, eluting with increasing proportions of MeOH in water.31. n.m.r. in solvent B: 1.54(br.s, 6H); 2.76(s, 3H); 3.56(m, 2H);4.07(s, 3H);4.02(d, 1H); 4.55(d, 1H); 5.15(d, 1H); 5.85(d, 1H); 7.06(s, 1H); 7.6(d,1H); 7.95(d, 1H).32. The starting material was prepared by the method described inFootnote 7,using 2-acetamido-1-mercaptoethane to give 3-fluoro-1-methyl-6-(2-acetamido-ethylthio)pyridazinium tetrafluoroborate as a paste.33. HPLC eluant MeOH/water/HOAc 25:74:1 v/v/v.34. n.m.r. in solvent B: 1.54(s, 6H); 1.83(s, 3H); 3.2-3.8(m, 6H); 4.0(s,3H);4.6(m, 2H); 5.19(d, 1H); 5.85(d, 1H); 7.0(s, 1H); 7.93(s, 2H).
EXAMPLES 172-178
The general process described in Example 1 was repeated using the appropriate 3-aminomethylcephalosporin derivative and the appropriate heterocycle. The following compounds were obtained:
__________________________________________________________________________ ##STR197##ExampleNo. R.sup.1 R.sup.2 R.sup.3 Yield Footnote__________________________________________________________________________172 CH.sub.3 H CH.sub.3 30 1, 2, 3173 CH.sub.2 CH.sub.2 Cl H CH.sub.3 24 1, 4, 5174 C(CH.sub.3).sub.2 CO.sub.2 H H ##STR198## 14 6, 7, 8175 " H CH.sub.2 CHCH.sub.2 18 9, 10, 11176 " H ##STR199## 23 12, 13, 14177 ##STR200## C.sub.2 H.sub.5 CH.sub.3 12 1, 15, 16178 ##STR201## H CH.sub.3 32 1, 17, 18__________________________________________________________________________Footnotes1. The starting material was 6-amino-3-fluoro-1-methylpyridaziniumtetrafluoroboratesee Example 13, Footnote 16.2. HPLC eluant MeOH/water/HOAc, 15:84:1 v/v/v.3. n.m.r. in solvent B: 3.25-3.8(m, 2H); 3.7(s, 3H); 4.0(s, 3H); 3.8(d,1H); 4.5(d, 1H);5.12(d, 1H); 5.76(d, 1H); 7.05(s, 1H); 7.25(s, 2H).4. HPLC eluant MeOH/water/HOAc, 20:79:1, v/v/v.5. n.m.r. in solvent B: 3.55(m, 2H); 3.7(s, 3H); 3.9-4.4(m, 6H); 5.15(d,1H); 5.76(d, 1H);7.05(s, 1H); 7.3(s, 2H).6. The starting material may be prepared as follows: 3,6-difluoropyridazine was mixedwith a solution of ammonia (10 equivalents) in ethanol, and heated in asealed tubefor 3 hours at 80-85.degree. C. The solvent was evaporated and6-amino-3-fluoropyri-dazine was extracted from the residue with EtOAc in a Soxhlet apparatusfor 3.5 hours.This compound in EtOH was further treated with p-nitrobenzyl bromide for2.5 hoursat 40.degree. C. The solvent was evaporated and the residue was washedtwice with anhydrousether, to give 6-amino-3-fluoro-1-(4-nitrobenzyl)pyridazinium bromide.7. HPLC eluant MeOH/water/HOAc, 35:64:1, v/v/v.8. n.m.r. in solvent B: 1.57(br.s., 6H); 3.5(m, 2H); 4.0(d, 1H); 4.45(d,1H); 5.15(d, 1H);5.45(br.s, 2H); 5.87(d, 1H); 7.1(s, 1H); 7.47(s, 2H); 7.65(d, 2H);8.35(d, 2H).9. The starting material may be prepared as follows: 6-amino-3-fluoropyridazine (2.5mmoles; Footnote 6), and 2 ml of allyl bromide in the minimum ofnitromethane wereheated to 60.degree. C. for 3 hours. Evaporation of the solvent, washingof the residuewith ether and drying under vacuum over P.sub.2 O.sub.5 gave1-allyl-6-amino-3-fluoropyridazinium bromide.10. HPLC eluant MeOH/water/HOAc, 30:69:1, v/v/v.11. n.m.r. in solvent B: 1.6(br.s, 6H); 3.57(m, 2H); 3.8-4.9(m, 4H);5.1-5.45(m, 3H);5.85(m, 2H); 7.17(s, 2H); 7.4(s, 2H).12. The starting material was prepared as follows: 6-amino-3-fluoropyridazine (1.15mmoles; Footnote 6) and alpha'-chloro-alpha,alpha,alpha-trifluoro-m-xylene in theminimum of DMF were heated to 60.degree. C. for 4 hours. Evaporation ofthe solventand trituration of the residue with anhydrous either gave6-amino-3-fluoro-1-(3-trifluoromethylbenzyl)-pyridazinium chloride.13. HPLC eluant MeOH/water/HOAC, 45:54:1, v/v/v.14. n.m.r. in solvent B: 1.54(s, 6H); 3.5(m, 2H); 4.03(d, 1H); 4.45(d,1H); 5.11(d, 1H);5.43(d, 1H); 5.9(d, 1H); 7.1(s, 1H); 7.45(s, 2H); 7.62-7.95(m, 4H).15. HPLC eluant MeOH/water/HOAc, 25-30:74-69:1 v/v/v.16. n.m.r. in solvent B: 1.07(t, 3H); 1.9-3.0(m, 6H); 3.46(m, 4H);4.25(d, 1H); 4.6(d,1H); 5.2(d, 1H); 5.86(d, 1H); 6.98(s, 1H); 7.4(d, 1H); 7.76(d, 2H).17. HPLC eluant MeOH/water/HOAc, 25-30:74-69:1 v/v/v.18. n.m.r. in solvent B: 1.72(m, 4H); 2.15(m, 4H); 3.55(m, 2H); 3.7(s,3H); 4.05(d, 1H);4.55(d, 1H); 5.15(d, 1H); 5.83(d, 1H); 7.06(s, 1H); 7.3(s, 2H).
EXAMPLES 179-180
The process described in Example 1 was repeated, using the appropriate heterocycle as starting material, to give the following compounds:
______________________________________ ##STR202##ExampleNo. R Yield % Footnotes______________________________________179 ##STR203## 15 1, 2, 3, 4180 ##STR204## 9 5, 6, 7, 8______________________________________1. The starting material was obtained by reaction of3-chloro-5,6,7,8-tetrahydrocinnoline (see Journal of Org.Chemistry 1955, Vol. 20, p.707) with trimethyloxonium tetra-fluoroborate in CH.sub.2 Cl.sub.2 overnight.2. The condensation was performed in water at pH 7.6(ambient temperature) with NaHCO.sub.3 as base.3. HPLC eluant, MeOH/water/HOAc, 30-40:69-59:1, v/v/v.4. n.m.r. in solvent B: 1.54(s, 6H); 1.86(m, 4H); 2.81(m, 4H);3.60(br.s, 2H); 3.96(s, 3H); 4.55(br.s, 2H); 5.16(d, 1H);5.88(d, 1H); 7.04(s, 1H); 7.73(s, 1H).5. The starting heterocycle may be prepared as follows: To astirred solution of 7.7-dichlorobicyclo-[3,2,0]heptan-6-one(see Tetrahedron, 1971, Vol. 27, p.615)(52.7 mmoles) inMeOH(25 ml) at 0.degree. C. was added dropwise hydrazine hydrate(3.3 ml.). Stirring was continued at 0.degree. C. for 0.75 hours,the solvent was evaporated, the residue was taken up in waterand extracted with CH.sub.2 Cl.sub.2, and the organic phaseswere combined, washed with water and dried (MgSO.sub.4).Evaporation of the solvent gave cis-1,1-dichloromethylcyclo-pentane carbohydrazide (yield 54%) having the followingn.m.r. in CDCl.sub.3 : 2.0(m, 6H); 2.8(m, 2H); 3.92(br.s,2H); 5.96(d, 1H); 7.0(br.s, 1H). The above hydrazide(28.7 mmoles) was dissolved in aqueous EtOH (40 ml.; 3:1 v/v)and heated at reflux for 1.75 hours. The reaction mixturewas diluted with water and extracted withCH.sub.2 Cl.sub.2. The combined CH.sub.2 Cl.sub.2 extractswere washed with water and dried (MgSO.sub. 4), the solvent wasevaporated to a small volume and ether was added to give4,5-dihydro-4,5-trimethylenepyridazin-3(2H)-one (yield 87%).N.m.r. in CDCl.sub.3 : 1.5-2.4(m, 6H); 2.6-3.0(m, 2H);6.95(d, 1H); 7.7(br.s, 1H). This dihydropyridazinone derivative(17.7 mmoles) was dissolved in HOAc (15 ml.) and bromine(17.6 mmoles) was added slowly with stirring. The reactionmixture was then heated 0.4 hours at 100.degree. C., cooled andthe HOAc evaporated. The residue was taken up in water andthe pH adjusted to 10 with 12N NaOH. After stirring for 3hours at room temperature, the reaction mixture was extractedwith CH.sub.2 Cl.sub.2, the combined CH.sub.2 Cl.sub.2extracts were washed with water, dried (MgSO.sub.4), andthe solvent was evaporated. The crude product was applied toa column of Kieselgel 60 (120 g). Elution with ether-MeOH,5-7:95-93 v/v, gave 4,5-trimethylenepyridazin-3(2H)-one in 24% yield. N.m.r. in CDCl.sub.3 : 2.13(q, 2H);2.91(t, 4H); 7.78(s, 1H). The above pyridazinone derivative(13.5 mmoles) was treated with 15 ml. of phosphorous oxychlorideat 80.degree. C. for 2 hours. After evaporation to dryness, theresidue was taken up in water, neutralised with NaHCO.sub.3and extracted with CH.sub.2 Cl.sub.2. The CH.sub.2 Cl.sub.2was evaporated and the residue was applied to a column ofKieselgel 60 (50 g). Elution with ether/MeOH, 95:5 v/v, gave3-chloro-4,5-trimethylenepyridazine (yield 42%). N.m.r. inCDCl.sub.3 : 2,2(q, 2H); 3.06(t, 2H); 3.08(t, 2H);9.02(s, 1H). This compound was then treated withtrimethyloxonium tetrafluoroborate in CH.sub.2 Cl.sub.2at ambient temperature for 0.5 hour to give a 9:1 mixtureof 1-methyl and 2-methyl-4,5-trimethylenepyridaziniumtetrafluoroborate in 90% yield. N.m.r. in CDCl.sub.3 ofthe 1-methyl derivative is: 2.39(q, 2H); 3.26(t, 2H);3.43(t, 2H); 4.59(s, 3H); 9.51(s, 2H).6. The condensation was performed in DMF/water at60.degree. C. for 6 hours in the presence of NaHCO.sub.3.7. HPLC eluant: MeOH/water/HOAc, 25:74:1, v/v/v.8. n.m.r. in solvent B: 1.51(s, 3H); 1.54(s, 3H); 2.17(m, 2H);2.9(m, 4H); 3.52(m, 2H); 4.21(s, 3H); 4.67(m, 1H); 4.84(m, 1H);5.1(d, 1H); 5.82(d, 1H); 7.05(s, 1H); 8.95(s, 1H).
EXAMPLES 181-191
The general process described in Example 1 was repeated using the appropriate 3-aminomethyl-cephalosporin derivative and the appropriate heterocycle. The following compounds were obtained:
__________________________________________________________________________ ##STR205##Example No. R.sup.1 R.sup.2 R.sup.3 Yield % Footnote__________________________________________________________________________181 C(CH.sub.3).sub.2 CO.sub.2 H H ##STR206## 13 1, 2, 3182 C(CH.sub.3).sub.2 CO.sub.2 H H ##STR207## 20 4, 5, 6183 " Et ##STR208## 24 4, 7, 8184 ##STR209## Et ##STR210## 31 4, 9, 10185 ##STR211## H ##STR212## 60 4, 11, 12186 C(CH.sub.3).sub.2 CO.sub.2 H H ##STR213## 27 13, 14, 15187 " Et ##STR214## 36 13, 16, 17188 ##STR215## H ##STR216## 54 13, 18, 19189 C(CH.sub.3).sub.2 CO.sub.2 H H ##STR217## 35 20, 21, 22190 ##STR218## H ##STR219## 23, 24, 25191 " H ##STR220## 26, 27, 28__________________________________________________________________________1. 6-Fluoro-2,3-dihydro-1H-imidazo(1,2,3)-pyridazinium bromide wasprepared according to theprocedure described in Croat. Chem. Acta 41, 135 (1969) using3,6-difluoropyridazine as startingmaterial. The hydrobromide salt in MeOH was further treated withdiazomethane to liberate thefree base, 6-fluoro-2,3-dihydroimidazo(1,2-b)pyridazine. Upon evaporationof the MeOH, thecompound was dissolved in CH.sub.2 Cl.sub.2 /nitromethane (5:3 v/v) andtrimethyloxoniumtetrafluoroborate was added to the reaction mixture. After 4 hours atambient temperature,the mixture was filtered, and the filtrate was evaporated to give6-fluoro-2,3-dihydro-1-methylimidazo(1,2-b)pyridazinium tetrafluoroborate.2. HPLC eluant, MeOH/water/HOAc, 25:74:1 v/v/v.3. n.m.r. in solvent B: 1.54(s, 6H); 2.99(s, 3H); 3.2-4.6(m, 8H); 5.13(d,1H); 5.82(d, 1H);7.05(s, 1H); 7.4(d, 1H); 7.5(d, 1H).4. The starting material was 6-chloro-thiazolo-[3,2-b]pyridaziniumperchlorate,(see J. Org Chem 34, 996, (1969).5. HPLC eluant, MeOH/water HOAc, 20:79:1, v/v/v.6. n.m.r. in solvent B: 1.55(s, 6H); 3.3(d, 1H); 3.55(d, 1H); 3.95(d,1H); 4.4(d, 1H);5.15(d, 1H); 5.55(d, 1H); 7.45(d, 1H); 8.47-8.6(m, 3H).7. The product was purified by chromatography on Diaion HP20 resin,eluting with increasingproportions of CH.sub.3 CN in water.8. n.m.r. in solvent B: 1.16(br t, 3H); 1.51(s, 6H); 3.5(m, 2H); 3.6(m,2H); 4.45(d, 1H);3.65(d, 1H); 5.18(d, 1H); 5.85(d, 1H); 7.0(s, 1H); 7.9(d, 1H); 8.54(d,1H); 8.66(d, 1H);8.7(d, 1H).9. HPLC eluant, MeOH/water/HOAc, 20-30:79-69:1 v/v/v.10. n.m.r. in solvent B: 1.15(br t, 3H); 1.91-2.4(m, 6H); 3.6(m, 4H);4.5(d, 1H);4.8(d, 1H); 5.17(d, 1H); 5.85(d, 1H); 6.98(s, 1H); 7.85(d, 1H); 8.65(d,1H); 8.5(d, 1H);8.6(d, 1H).11. HPLC eluant, MeOH/water/HOAc, 30:69:1, v/v/v.12. n.m.r. in solvent B: 1.7(m, 4H); 2.1(m, 4H); 3.5(d, 1H); 3.7(d, 1H);4.1(d, 1H); 4.6(d, 1H);5.14(d, 1H); 5.84(d, 1H); 7.04(s, 1H); 7.45(d, 1H); 8.6(d, 1H); 8.47(d,1H); 8.52(d, 1H).13. The starting material was 6-chloro-3-methylthiozolo[3,2-b]pyridazinium perchlorate(see J. Org. Chem. 34, 996(1964)).14. HPLC eluant, MeOH/water/HOAc, 25:74:1, v/v/v.15. n.m.r. in solvent B: 1.52(s, 3H); 1.54(s, 3H); 3.55(m, 2H); 4.1(d,1H); 4.65(d, 1H);5,15(d, 1H); 5.85(d, 1H); 7.0(s, 1H); 7.47(d, 1H)); 8.62(d, 1H); 8.23(s,1H).16. HPLC eluant, MeOH/water/HOAc, 30:69:1, v/v/v.17. n.m.r. in solvent B: 1.15(br.t, 3H); 1.5(br.s, 6H); 2.51(s, 3H);3.4(q, 2H); 3.65(m, 2H);4.5(d, 1H); 4.8(d, 1H); 5.17(d, 1H); 5.87(d, 1H); 7.0(s, 1H); 7.87(d,1H); 8.64(d, 1H); 8.2(s, 1H).18. HPLC eluant MeOH/water/HOAc, 30:69:1, v/v/v.19. n.m.r. in solvent B: 1.7(m, 4H); 2.1(m, 4H); 2.52(s, 3H); 3.56(d,1H); 3.6(d, 1H); 4.3(d, 1H);4.6(d, 1H); 5.15(d, 1H); 5.86(d, 1H); 7.04(s, 1H); 7.47(d, 1H); 8.61(d,1H); 8.61(d, 1H); 8.22(s, 1H).20. The starting material was prepared by treating 6-fluoroimidazo(1,2-b)pyridazine, (obtained bya process described in Tetrahedron, 23, 387 (1967), with trimethyloxoniumtetrafluoroborate inCH.sub.2 Cl.sub.2 at ambient temperature for 5 hours. The precipitate wasfiltered, to give6-fluoro-1-methylimidazo(1,2-b)pyridazinium tetrafluoroborate.21. HPLC eluant, MeOH/water/HOAc, 20:79:1, v/v/v.22. n.m.r. in solvent B: 1.56(br.s, 6H); 3.61(m, 2H); 3.98(s, 3H); 4.1(d,1H); 4.6(d, 1H); 5.15(d, 1H);5.83(d, 1H); 7.02(s, 1H); 7.31(d, 1H); 8.04(d, 1H); 8.09(d, 1H); 8.21(d,1H).23. For starting material see Example 229, Footnote 2.24. HPLC eluant was MeOH/water/HOAc, 25:74:1, v/v/v.25. n.m.r. in solvent B: 1.7(m, 4H); 2.14(m, 4H); 3.48(d, 1H); 3.7(d,1H); 4.15(s, 3H); 4.14(d, 1H);4.45(d, 1H); 5.14(d, 1H); 5.85(d, 1H); 7.05(s, 1H); 7.52(d, 1H); 8.35(d,1H); 9.47(s, 1H).26. For starting material, see Example 222, Footnote 5.27. HPLC eluant was MeOH/water/HOAc, 30:69:1, v/v/v.28. n.m.r. in solvent B: 1.7(m, 4H); 2.15(m, 4H); 2.44(d, 1H); 2.65(d,1H); 4.05(s, 3H); 4.55(d, 1H);4.95(d, 1H); 5.15(d, 1H); 5.82(d, 1H); 7.05(s, 1H); 7.92(s, 2H); 8.9(s,1H).
EXAMPLE 192 ##STR221##
The starting material was 3-fluoro-6-methoxy-1-methyl-pyridazinium tetrafluoroborate, which was obtained by treatment of 6-fluoro-2-methylpyridazin-3-one with trimethyloxonium tetrafluoroborate in CH.sub.2 CL.sub.2 at ambient temperature.
The condensation reaction was carried out in DMF/water in the presence of NaHCO.sub.3 at ambient temperature for 0.5 hours. Yield 12%.
HPLC eluant, MeOH/water/HOAc, 25:74:1, v/v/v.
n.m.r. in solvent B: 1.54 (s, 6H); 3.6 (br.s, 2H); 3.93 (s, 3H); 3.45 (d, 1H); 3.85 (d, 1H), 5.16 (d, 1H); 5.88 (d, 1H); 7.04 (s, 1H); 7.98-8.17 (m, 2H).
EXAMPLE 193 ##STR222##
This compound was obtained as follows: To 5-amino-3-chloropyridazine (see Yakugaku Zasshi 82, 857-60, (1962); Chem. Abstracts 59, 1631j) in nitromethane was added trimethyloxonium tetrafluoroborate. After 3 hours at ambient temperature the solvent was evaporated, and the ether was added to give a mixture of 5-amino-3-chloro-1-methylpyridazinium and 5-amino-3-chloro-2-methylpyridazinium tetrafluoroborates, which were used without further separation. The standard procedure was used for the condensation, which was carried out in a DMF/water mixture in the presence of NaHCO.sub.3 at 70.degree. C. for 1.25 hours. The final compound was worked up by HPLC using MeOH/water/HOAc, 22.5:76.5:1 v/v/v, as eluant. Yield 25%. n.m.r. of the single isomer product in solvent B: 1.53 (br.s, 6H); 3.59 (m, 2H); 3.83 (s, 3H); 4.56 (m, 2H); 5.15 (d, 1H); 5.9 (d, 1H); 6.23 (d, 1H); 7.0 (s, 1H); 7.92 (d, 1H).
EXAMPLES 194-195
The general process described in Example 1 was repeated, using the appropriate 3-aminomethylcephalosporin derivative and the appropriate heterocycle, to give the following compounds:
______________________________________ ##STR223##Example No. R Yield % Footnotes______________________________________194 ##STR224## 37 1, 2, 3195 ##STR225## 23 4, 5, 6______________________________________Footnotes:1. For the quaternary heterocyclic starting material, see Example182, Footnote 4.2. HPLC eluant, MeOH/water/HOAc, 20-25:79-74:1, v/v/v.3. n.m.r. in solvent B: 1.48(s, 6H); 3.45(d, 1H); 3.71(d, 1H);4.11(d, 1H); 4.57(d, 1H); 5.12(d, 1H); 5.84(d, 1H); 7.46(d, 1H);8.47(d, 1H); 8.57(d, 2H).4. For the quarternary heterocyclic starting material, see Example13, Footnote 6.5. HPLC eluant, MeOH/water/HOAc, 20:79:1, v/v/v.6. n.m.r. in solvent B: 1.47(s, 6H); 3.5(m, 2H); 3.6(s, 3H);3.91(d, 1H); 4.42(d, 1H); 5.09(d, 1H); 5.83(d, 1H); 7.26(s, 2H).7. The 3-aminomethyl cephalosporin starting material was obtainedby adding the corresponding known 3-azidomethyl cephalosporinderivative to 90% v/v aqueous TFA and stirring for 3 hours atambient temperature. After cooling at 0.degree., Raney nickel(wet 2.0 g) was added and the mixture was stirred for 0.5 hours at0.degree. and 1 hour at ambient temperature. The mixture was filtered,the filtrate evaporated and the crude product purified bychromatography on a Diaion HP 20 column, eluting withincreasing proportions of acetonitrile in water. Appropriatefractions were combined, evaporated to a small volume andfreez-dried to give the required compound (0.81 g; yield 43%)having the following n.m.r. in solvent B: 1.49(s, 6H);3.5-4.0(m, 4H); 5.18(d, 1H); 5.9(d, 1H).
EXAMPLES 196-218
The general procedure described in Examples 1-4 was used, except that the reaction was carried out at ambient temperature for 5 hours before quenching with HOAc. The appropriate 3-aminomethylcephalosporin was used, the reaction was worked up by chromatography on Diaion CHP20P resin to purify. The following compounds were made:
__________________________________________________________________________ ##STR226##Example No. R.sup.1 R.sup.2 Yield % Footnotes__________________________________________________________________________196 CH.sub.2 CO.sub.2 H ##STR227## 46 1, 2197 CH.sub.2 CO.sub.2 H ##STR228## 40 3, 4198 ##STR229## ##STR230## 34 1, 5, 6199 ##STR231## ##STR232## 40 1, 6, 7200 ##STR233## ##STR234## 28 3, 6, 8201 ##STR235## ##STR236## 22 3, 6, 9202 ##STR237## ##STR238## 29 3, 10203 ##STR239## ##STR240## 20 11, 12204 CH.sub.2 CONHCH.sub.3 ##STR241## 67 1, 13, 14205 CH.sub.2 CON(CH.sub.3).sub.2 ##STR242## 64 1, 15, 16206 CH.sub.2 CONHCH.sub.2 CF.sub.3 ##STR243## 57 1, 17, 18207 ##STR244## ##STR245## 69 1, 19, 20208 C.sub.2 H.sub.5 ##STR246## 18 11, 21209 ##STR247## ##STR248## 60 1, 22, 23210 ##STR249## ##STR250## 46 1, 23, 24211 ##STR251## ##STR252## 33 3, 25212 ##STR253## ##STR254## 69 26, 27213 ##STR255## ##STR256## 57 28, 29214 ##STR257## ##STR258## 56 30, 31215 ##STR259## ##STR260## 53 30, 32216 ##STR261## ##STR262## 61 30, 33217 ##STR263## ##STR264## 42 34, 35218 ##STR265## ##STR266## 52 1, 36__________________________________________________________________________Footnotes1. Starting material was 3-chloro-1-methylpyridazinium iodide.2. n.m.r. in solvent A: 3.36(d, 1H); 3.59(d, 1H); 4.08(d, 1H); 4.24(s,3H); 4.48(d, 1H); 4.53(s, 2H);5.03(d, 1H); 5.71(d, 1H); 6.8(s, 1H); 7.6(m, 1H); 7.88(m, 1H); 8.88(m,1H).3. Starting material was 7-methylthiothiazolo-[3,2-a]pyrimidiniumtetrafluoroborate.4. n.m.r. in solvent A: 3.35(d, 1H); 3.6(d, 1H); 4.25(d, 1H); 4.51(s,2H); 4.64(d, 1H); 5.03(d, 1H);5.73(d, 1H); 6.81(s, 1H); 7.04(d, 1H); 7.65(d, 1H); 8.08(d, 1H); 8.79(d,1H).5. n.m.r. in solvent A: 1.4(d, 3H); 3.39(d, 1H); 3.61(d, 1H); 4.08(d,1H); 4.23(s, 3H); 4.53(d, 1H);4.6(q, 1H); 5.06(d, 1H); 5.8(d, 1H); 6.81(s, 1H); 7.6(m, 1H); 7.93(m,1H); 8.9(m, 1H).Retention time 3.6 minutes on a reversed phase Partisil 10/25 ODS-2column, usingMeOH/H.sub.2 O/TFA, 35:65:0.2, v/v/v as eluant.6. The 3-aminomethylcephalosporin may be prepared as follows: To astirred suspension of 2-(2-formamidothiazol-4-yl)-2-oxoacetic acid (6.1 g) in DMF (50 ml) wasadded pyridine (2.65 ml)and 4N aqueous hydrochloric acid (8.52 ml.), followed by dropwiseaddition of t-butyl 2-amino-oxypropionate (5.28 g). After stirring for 18 hours at ambienttemperature, the mixture wasdiluted with water, acidified, and worked up in a conventional manner byextraction into EtOAc.Crude product was dissolved in a mixture of ether (80 ml) and EtOAc (40ml), and N-methyl-morpholine (7.6 ml) added. The precipitate was filtered to give2-[(Z)-1-(tert-butoxy-carbonyl)ethoxy-imino]-2-(2-formamidothiazol-4-yl)acetic acid as itsN-methyl-morpholine salt,having the following n.m.r. in d.sub.6 DMSO: 1.35(d, 3H); 1.41(s, 9H);2.35(s, 3H); 2.56(t, 4H);3.65(t, 4H); 4.55(q, 1H); 7.35(s, 1H); 8.46(s, 1H).Oxalyl chloride (2.61 ml) and DMF (2.3 ml) were added to CH.sub.2Cl.sub.2 (100 ml) at -10.degree. C.,and stirred for 30 minutes. The above N-methylmorpholine salt (12.06 g)was added, followed byN-methylmorpholine (0.60 ml), and stirring continued for 30 minutes.Meanwhile, in a separateflask, a suspension of 7-amino-3-azidomethyl-ceph-3-em-4-carboxylic acid(7.27 g) in CH.sub.2 Cl.sub.2(40 ml) was treated with N,O-bis(trimethylsilyl)acetamide (14.1 ml), andstirred for 1 hour togive a clear solution, which was transferred by syringe to the acidchloride solution. Afterstirring for 1 hour at -10.degree. C., the temperature was allowed tocome to ambient. Themixture was poured into water (100 ml) and organics extracted into EtOAc.Removal of solventgave 3-azido-methyl-7-[ 2-((Z)-1-(t-butoxycarbonyl)ethoxyimino)-2-(2-formamidothiazol-4-yl)-acetamido]ceph-3-em-4-carboxylic acid, as a mixture of diastereoisomers,having the followingn.m.r. in solvent A: 1.35(s, 9H); 1.37 and 1.38(2xd, 3H); 3.41(2xd, 1H);3.61(d, 1H); 3.82(2xd, 1H);4.34(d, 1H); 4.56(2xq, 1H); 5.14(2xd, 1H); 5.83(d, 1H); 7.32(2xs, 1H);8.39(s, 1H).A solution of the above 3-azidomethyl compound (16.5 g) in a mixture ofisopropanol (110 ml),water (20 ml) and concentrated hydrochloric acid (7.5 ml) was stirred for3 days at ambienttemperature. After cooling to 0.degree. C., the pH was raised to 3 usingtriethylamine, solvents wereevaporated, the residue treated with water (100 ml.) and organicsextracted into EtOAc. Evapor-ation gave 7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-(t-butoxycarbonyl)ethoxyimino)acetamido]-3-azidomethylceph-3-em-4-carboxylic acid, as a mixture of diastereoisomers,having the followingn.m.r in solvent A: 1.4 and 1.42(2xc, 3H); 1.44(s, 9H); 3.46(2xd, 1H);3.66(d, 1H); 3.88(2xd, 1H);4.39(d, 1H); 4.55(q, 1H); 5.18(d, 1H); 5.84(d, 1H); 6.74(2xs, 1H).The above deprotected 3-azidomethylcephem (12.2 g) was added to 90% v/vaqueous TFA (60ml) pre-cooled to 0.degree. C., and stirred for 30 minutes. Raney nickel(wet, 2.5 g) was added, and themixture stirred 20 minutes, allowing the temperatre to rise to ambient.The mixture was filtered,solvents evaporated and the residue dissolved in water, and the pH raisedto 3 with sodiumacetate. Careful chromatography on Diaion HP20SS, eluting successivelywith water, thenwater/CH.sub.3 CN, 97.5:2.5 v/v and water/CH.sub.3 CN 95:5 v/v, allowedsubstantial separation of thetwo diastereoisomers of 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxyethoxy-imino)acetamido]ceph-3-em-4-carboxylic acid. The more polar (m.p.)isomer, having retentiontime 3.05 minutes on a reversed phase Partisil PXS 10/25 ODS-2 column,using MeOH/H.sub.2 O/HOAc 20:80:1 v/v/v as eluant, had the following n.m.r. in solvent A:1.37(d, 3H); 3.23(d, 1H);3.44(d, 1H); 3.53(d, 1H); 3.65(d, 1H); 4.52(q, 1H); 4.99(d, 1H); 5.8(d,1H); 6.79(s, 1H).The less polar (l.p.) isomer, having retention time 4.7 minutes underidentical conditions to theabove, had the following n.m.r. in solvent A: 1.43(d, 3H); 3.2(d, 1H);3.42(d, 1H); 3.53(d, 1H);3.64(d, 1H); 4.52(q, 1H); 5.0(d, 1H); 5.72(d, 1H); 6.81(s, 1H).7. n.m.r. in solvent A: 1.42(d, 3H); 3.45(d, 1H); 3.64(d, 1H); 4.08(d,1H); 4.24(s, 3H); 4.56(d, 1H);4.63(q, 1H); 5.12(d, 1H); 5.81(d, 1H); 6.79(s, 1H); 7.6(m, 1H); 7.95(m,1H); 8,95(m, 1H). Retentiontime 4.6 min. on a reversed phase Partisil PXS 10/25 ODS-2 column, usingMeOH/H.sub.2 O/TFA,35:65:0.2 v/v/v, as eluant.8. n.m.r. in solvent A: 1.38(d, 3H); 3.31(d, 1H); 3.59(d, 1H); 4.27(d,1H); 4.56(q, 1H); 4.59(d, 1H);5.02(d, 1H); 5.77(d, 1H); 6.78(s, 1H); 7.05(d, 1H); 7.66(d, 1H); 8.1(d,1H); 8.69(d, 1H). Retentiontime 3.3 minutes on a reversed phase Partisil 10/25 ODS0-2 column, usingMeOH/H.sub.2 O/TFA,40:60:0.2, v/v/v, as eluant.9. n.m.r. in solvent A: 1.41(d, 3H); 3.36(d, 1H); 3.61(d, 1H); 4.23(d,1H); 4.6(m, 3H); 5.05(d, 1H);5.74(d, 1H); 6.8(s, 1H); 7.03(d, 1H); 7.67(d, 1H); 8.11(d, 1H); 8.7(d,1H). Retention time 4.3minutes on a reversed phase Partisil 10/25 ODS-2 column, usingMeOH/H.sub.2 O/TFA, 40:60:0.2,v/v/v, as eluant.10. n.m.r. in solvent A: 2.38(t, 4H); 3.35(d, 1H); 3.62(d, 1H); 4.27(d,1H); 4.66(d, 1H); 5.06(d, 1H);5.76(d, 1H); 6.76(s, 1H); 7.04(d, 1H); 7.66(d, 1H); 8.1(d, 1H); 8.7(d,1H).11. The basic procedure of Examples 1-4 was used, except the reactionmixture was heated at60.degree. C. for 2 hours, and product was isolated by chromatography onDiaion HP20SS. The startingheterocycle was 6-amino-3-fluoro-1-methylpyridazinium tetrafluoroborate.12. n.m.r. in solvent A: 2.33(m, 4H); 3.28(d, 1H); 3.56(d, 1H); 3.65(s,3H); 3.83(d, 1H); 4.27(d,1H); 5.0(d, 1H); 5.68(d, 1H); 6.7(s, 1H); 8.23(s, 2H).13. n.m.r. in solvent A: 2.66(s, 3H); 3.35(d, 1H); 3.61(d, 1H); 4.08(d,1H); 4.26(s, 3H); 4.47(s, 2H);4.51(d, 1H); 5.03(d, 1H); 5.72(d, 1H); 6.87(s, 1H); 7.62(m, 1H); 7.94(m,1H); 8.91(d, 1H).14. The 3-aminomethylcephalosporin may be prepared as follows: Using thegeneral procedure ofFootnote 6 above, 2-((Z)-N-methylcarbamoylmethoxyimino)-2-(2-tritylaminothiazol-4-yl)aceticacid was condensed with 7-amino-3-azidomethylceph-3-em-4-carboxylic acidto give theappropriate 7-amido-3-azidomethylcephalosporin. This was then reduced,with accompanyingremoval of the trityl group, by the same general procedure to give3-aminomethyl-7-[2-(2-amino-thiazol-4-yl)-2-((Z)-N-methylcarbamoylmethoxyimino)acetamido]ceph-3-em-4-carboxylic acid,used without further characterisation in subsequent stages.15. n.m.r. in solvent A: 2.8(s, 3H); 2.92(s, 3H); 3.3(d, 1H); 3.61(d,1H); 4.06(d, 1H); 4.24(s, 3H);4.49(d, 1H); 4.81(s, 2H); 5.02(d, 1H); 5.71(d, 1H); 6.82(s, 1H); 7.6(m,1H); 7.93(dd, 1H); 8.89(d,1H).16. The 3-aminomethylcephalosporin may be prepared as follows: Using thegeneral procedure ofFootnote 6 above, 2-((Z)-N,N-dimethylcarbamoylmethoxyimino)-2-(2-tritylaminothiazol-4-yl)acetic acid was condensed with 7-amino-3-azidomethylceph-3-em-4-carboxylic acid, to givethe appropriate 7-amido-3-azidomethylcephalosporin. This was reduced, bythe same generalprocedure, to give, after chromatography on Diaion HP20SS resin andeluting with isopropanol/water, 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2((Z)-N,N-dimethylcarbamoylmethoxy-imino)acetamido]ceph-3-em-4-carboxylic acid, used without furthercharacterisation forsubsequent stages.17. n.m.r. in solvent A: 3.34(d, 1H); 3.63(d, 1H); 3.94(m, 2H); 4.08(d,1H); 4.25(s, 3H); 4.55(d,1H); 4.6(s, 2H); 5.05(d, 1H); 5.74(d, 1H); 6.87(s, 1H); 7.63(m, 1H);7.94(dd, 1H); 8.92(d, 1H).18. The starting material may be obtained as follows: 2,2,2-Trifluoroethylamine hydrochloride(27.1 g) was suspended in dichloromethane (300 ml), cooled to 5.degree.,and triethylamine (55.6 ml)was run in. A solution of bromoacetyl bromide (18 ml) in dichloromethane(200 ml) was run inover the course of 3 hours. After a conventional extractive work-up, thecrude product wasdistilled to give N-(2,2,2-trifluoroethyl)-2-bromoacetamide, n.m.r. inCDCl.sub.3 : 3.93(m, 4H);6.9(br, 1H).Ethyl 2-(Z)-hydroxyimino-2-(2-tritylaminothiazol-4-yl)acetatehydrochloride (19.7 g) wasdissolved in DMSO (150 ml), treated with potassium carbonate (15.4 g),and the above bromo-acetamide (12.7 g) was added. After stirring for 24 hours, the reactionmixture was diluted withwater (500 ml) and worked-up by conventional extraction. Product waspurified by chromato-graphy on silica gel, to give ethyl 2-[(Z)-N-(2,2,2-trifluoroethyl)carbamoylmethoxyimino]-2-(2-tritylaminothiazol-4-yl)acetate, n.m.r. in CDCl.sub.3 : 1.38(t, 3H);3.94(m, 2H); 4.41(q, 2H), 4.82(s,2H); 6.63(s, 1H); 7.0 (m, 2H); 7.35(m, 15H).The ester was dissolved in methanol, and hydrolysed at ambienttemperature by addition ofaqueous sodium hydroxide. The sodium salt precipitated and was filteredoff. After resuspensionin water and acidification with 2N hydrochloric acid, there was obtained2-[(Z)-N-(2,2,2-tri-fluoroethyl)carbamoylmethoxyimino]-2-(2-tritylaminothiazol-4-yl)aceticacid. Following theprocedure outlined in Footnote 14 above, this was condensed with7-amino-3-azidomethylceph-3-em-4-carboxylic acid, and the product was reduced, to give3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-[(Z)-N-(2,2,2-trifluoroethyl)carbamoylmethoxyimino]acetamido]-ceph-3-em-4-carboxylicacid, used without further purification for subsequent stages.19. Product obtained as a 6:4 mixture of diastereoisomers, n.m.r. insolvent A: 2.26(m, 2H);3.22(m, 2H); 3.32(d, 1H); 3.59(d, 1H); 4.06(d, 1H); 4.25(s, 3H); 4.48(d,1H); 4.68 and 4.74(2xt,1H); 5.01(d, 1H); 5.66(d, 1H); 6.78 and 6.81(2xs, 1H); 7.61(br. 1H);7.93(dd, 1H); 8.91(d, 1H).20. The 3-aminomethylcephalosporin may be prepared as follows: Using thegeneral procedureoutlined in Footnote 6 above, 2-((Z,RS)-2-oxopyrrolidin-3-yloxy-imino)-2-(2-tritylaminothiazol-4-yl)acetic acid was condensed with 7-amino-3-azidomethylceph-3-em-4-carboxylic acid to give3-azidomethyl-7-[2-((Z,RS)-2-oxopyrrolidin-3-yloxyimino)-2-(2-tritylaminothiazol-4-yl)acetamido]ceph-3-em-4-carboxylic acid, n.m.r. in solvent A: 2.53(m,2H); 3.3-3.6(m, 4H);4.09(d, 1H); 4.45(d, 1H); 5.05(m, 2H); 5.88(d, 1H); 6.75(s, 1H); 7.33(m,15H).The azide was reduced by the standard procedure to give3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2((Z,RS)-2-oxopyrrolidin-3-yloxyimino)acetamido]-ceph-3-em-4-carboxylic acid as a 6:4mixture of diastereoisomers, n.m.r. in solvent A: 2.3(m, 2H); 3.1-3.8(m,6H); 4.73 and 4.78(2xt,1H); 5.02(d, 1H); 5.72(d, 1H); 6.81 and 6.83(2xs, 1H).21. n.m.r. in solvent A: 1.23(t, 3H); 3.32(d, 1H); 3.6(d, 1H); 3.71(s,3H); 4.05(m, 4H); 5.01(d, 1H);5.68(d, 1H); 6.71(s, 1H); 7.28(s, 2H).22. n.m.r. in solvent A: 0.93(t, 3H); 1.78(quintet, 2H); 3.34(d, 1H);3.58(d, 1H); 4.06(br, 1H);4.22(s, 3H); 4.42(t, 1H); 4.46(br, 1H); 5.02(d, 1H); 5.75(d, 1H);7.59(br, 1H); 7.91(br, 1H); 8.9(d,1H). Retention time 5.8 minutes on a reversed phase Partisil PXS 10/25ODS-2 column, usingMeOH/water/TFA 40:60:0.2 v/v/v as eluant.23. The 3-aminomethylcephalosporin may be prepared as follows: Hydrazinehydrate (15 ml) wasadded dropwise to a stirred solution of t-butyl 2-(N-phthalimido-oxy)-butyrate (45.8 g) indichloromethane (600 ml) at ambient temperature. After 2 hours, a mixtureof 25% aqueousammonia (120 ml) and water (200 ml) was added, and t-butyl-2-aminoxybutyrate isolated by aconventional extractive procedure. By following the procedure of Footnote6, this wasconverted to 2-[(Z)-1-(t-butoxycarbonyl)propoxyimino]-2-(2-formamidothiazol-4-yl)-acetic acid,precipitated as its triethylamine salt, having the following n.m.r. ind.sub.6 -DMSO: 0.94(t, 3H);1.21(t, 9H); 1.46(s, 9H); 1.73(quintet, 2H); 3.03(q, 6H); 4.3(t, 1H);7.25(s, 1H); 8.51(s, 1H).Following the procedure of Footnote 6, the above acetic acid wascondensed with 7-amino-3-azido-methylceph-3-em-4-carboxylic acid, the formyl group was removed byhydrolysis, and theresulting product was reduced to the 3-aminomethylcephem. Carefulchromatography on DiaionHP20SS, eluting successively with water, water/CH.sub.3 CN, 97.5:2.5 v/v,water CH.sub.3 CN, 95:5 v/vand water/CH.sub.3 CN, 92.5:7.5 v/v, allowed substantial separation ofthe diasteroisomers of3-aminomethyl-7-[2-aminothiazol-4-yl)-2-((Z)-1-carboxy-propoxyimino)acetamido]ceph-3-em-4-carboxylic acid. The more polar (m.p.) isomer, having retention time 3.02minutes on a reversedphase Partisil PXS 10/25 ODS-2 column, using MeOH/H.sub.2 O/HOAc 30:70:1v/v/v as eluant, hadthe following n.m.r. in solvent A: 0.95(t, 3H); 1.8(m, 2H); 3.28(d, 1H);3.43(d, 1H); 3.58(d, 1H);3.69(d, 1H); 4.45(t, 1H); 5.02(d, 1H); 5.75(d, 1H); 6.79(s, 1H).The less polar (l.p.) isomer, having retention time 3.76 minutes underidentical conditions to theabove, had the following n.m.r, in solvent A: 1.0(t, 3H); 1.83(m, 2H);3.28(d, 1H); 3.43(d, 1H);3.58(d, 1H); 3.69(d, 1H); 4.43(t, 1H); 5.03(d, 1H); 5.71(d, 1H); 6.77(s,1H).24. n.m.r. in solvent A: 0.94(t, 3H); 1.79(quintet, 2H); 3.34(d, 1H);3.56(d, 1H); 4.05(br, 1H);4.22(s, 3H); 4.37(t, 1H); 4.47(br, 1H); 5.03(d, 1H); 5.69(d, 1H); 6.77(d,1H); 7.6(br, 1H); 7.92(br,1H); 8.9(d, 1H)). Retention time 6.8 minutes on a reversed phase ParisilPXS 10/25 ODS-2column, using MeOH/water/TFA 40:60:0.2 v/v/v as eluant.25. n.m.r. in solvent A: 1.65(m, 4H); 2.03(m, 4H); 3.34(d, 1H); 3.61(d,1H); 4.25(d, 1H); 4.65(d,1H); 5.03(d, 1H); 5.72(d, 1H); 6.75(s, 1H); 7.03(d, 1H); 7.67(d, 1H);8.1(d, 1H); 8.7(d, 1H).26. Starting material was 2,3-dihydro-7-methylthiothiazolo[2,3-a]pyrimidinium tetrafluoroborate.27. n.m.r. in solvent A: 1.67(m, 4H); 2.04(m, 4H); 3.27(d, 1H); 3.58(d,1H); 3.69(t, 2H); 4.22(d,1H); 4.55(d, 1H); 4.6(t, 2H); 5.04(d, 1H); 5.73(d, 1H); 6.67(d, 1H);6.75(s, 1H); 8.12(d, 1H).28. The starting material was prepared by dissolving 1-(2-cyanothiazol-4-yl)methyl-4-methylthio-pyridinium chloride (242 mg) in a mixture of dichloromethane (11 ml) andTFA (2 ml), coolingin ice, and treating with 3-chloroperoxybenzoic acid (232 mg). Afterstirring for 40 minutessolvents were evaporated and the residue was treated with water (8 ml)and extracted with ether(3 portions of 5 ml). The aqueous layer containing 1-(2-cyanothiazol-4-yl)methyl-4-methyl-sulphinylpyridinium salts was used as such.29. n.m.r. in solvent A: 1.82(m, 2H); 2.37(t, 4H); 3.29(d, 1H); 3.52(d,1H); 4.21(d, 1H); 4.41(d,1H); 5.04(d, 1H); 5.53(s, 2H); 5.72(d, 1H); 6.75(s, 1H); 6.95(dd, 1H);7.34(dd, 1H); 8.17(dd, 1H);8.29(s, 1H); 8.34(dd, 1H).30. The starting material was prepared as in Footnote 28, but startingfrom 4-methylthio-1-(1,2,3-thiadiazol-4-yl)methylpyridinium methanesulphonate.31. n.m.r. in solvent A: 1.82(m, 2H); 2.38(t, 4H); 3.28(d, 1H); 3.51(d,1H); 4.2(d, 1H); 4.41(d, 1H);5.05(d, 1H); 5.71(d, 1H); 5.82(s, 2H); 6.73(s, 1H); 6.94(dd, 1H);7.35(dd, 1H); 8.25(d, 1H); 8.43(d,1H); 9.26(s, 1H).32. n.m.r. in solvent A: 1.41(d, 3H); 3.27(d, 1H); 3.51(d, 1H); 4.21(d,1H); 4.51(d, 1H); 4.59(q,1H); 5.06(d, 1H); 5.74(d, 1H); 5.86(s, 2H); 6.76(s, 1H); 6.95(dd, 1H);7.35(dd, 1H); 8.27(d, 1H);8.45(d, 1H); 9.27(s, 1H).Retention time 5.0 minutes on a reversed phase Partisil PXS 10/25 ODS-2column using MeOH/water/TFA 35:65:0.2 v/v/v as eluant.33. n.m.r. in solvent A: 0.92(t, 3H); 1.78(quintet, 2H); 3.27(d, 1H);3.36(d, 1H); 4.21(d, 1H);4.42(d, 1H); 4.44(t, 1H); 5.04(d, 1H), 5.72(d, 1H); 5.86(s, 2H); 6.76(s,1H); 6.95(dd, 1H); 7.32(dd,1H); 8.27(d, 1H); 8.45(d, 1H); 9.26(s, 1H). Retention time 7.6 minutes ona reversed phase PartisilPXS 10/25 ODS-2 column, using MeOH/water/TFA 35:65:0.2 v/v/v as eluant.34. The starting material was prepared as in Footnote 28, but startingwith 1-(E-3-cyano-2-methylallyl)-4-methylthiopyridinium bromide, and using two equivalents of3-chloroperoxy-benxoic acid, to give substantially 1-(E-3-cyano-2-methylallyl)-4-methanesulphonylpyridiniumbromide.35. n.m.r. in solvent A: 0.95(t, 3H); 1.8(quintet, 2H); 1.96(d, 3H);3.29(d, 1H); 3.54(d, 1H);4.24(d, 1H); 4.43(d, 1H); 4.45(t, 1H); 4.93(s, 2H); 5.06(d, 1H); 5.23(d,1H); 5.74(d, 1H); 6.78(s,1H); 6.96(dd, 1H); 7.31(dd, 1H); 8.01(dd, 1H); 8.19(dd, 1H).Retention time 4.3 minutes on a reversed phase Partisil PXS 10/25 ODS-2column, using MeOH/Water/TFA 40:60:0.2 v/v/v as eluant.36. n.m.r. in solvent A: 1.67(br, 4H); 2.0(br, 4H); 3.31(d, 1H); 3.61(d,1H); 4.24(s + m, 4H);4.46(d, 1H); 5.01(d, 1H); 5.71(d, 1H); 6.72(s, 1H); 7.6(m, 1H); 7.82(m,1H); 8.84(d, 1H).
EXAMPLE 219
The general procedure of Examples 7-14 was repeated using DMF as solvent at ambient temperature, with the appropriate starting materials. Products were worked up by chromatography on Diaion CHP20P resin, eluting with CH.sub.3 CN/water mixtures. The following compounds were made.
______________________________________ ##STR267##Example Yield Foot-No. R.sup.1 R.sup.2 % notes______________________________________219 ##STR268## ##STR269## 48 1, 2, 3______________________________________Footnotes1. Starting material was 1-methyl-4-methylthiothieno[2,3-d]pyrimidinium tetrafuloroborate.2. n.m.r. in solvent A: 1.29(t, 3H); 1.38(d, 3H); 3.21(d, 1H);3.44(d, 1H); 4.01(s + m, 5H); 4.56(q, 1H); 4.86(d, 1H); 5.05(d,1H); 5.22(d, 1H); 5.74(d, 1H); 6.75(s, 1H); 7.8(d, 1H); 7.9(d, 1H);8.81(s, 1H).Retention time 5.1 minutes on a reversed phase Partisil PXS 10/25ODS-2 column, using MeOH/H.sub.2 O/TFA 40:60:0.2 as eluant.3. The cephalosporin intermediate may be prepared as follows:To a stirred solution of triethylamine (0.18 ml) in MeOH (50 ml)was added 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxyethoxyimino)-acetamido]ceph-3-em-4-carboxylic acid(more polar isomer, 0.6 g), followed by sodium cyanoborohydride(0.08 g). A solution of acetaldehyde (0.07 ml) in MeOH (5 ml)was added over 40 minutes, then the solvent was removed and theresidue was dissolved in water (100 ml). After reducing the pHto 3.5, the solution was loaded onto a column of Diaion CHP20Presin. Elution with 95:5 H.sub.2 O:CH.sub.3 CN gave 7-[2-(2-amino-thiazol-4-yl)-2-((Z)-1-carboxyethoxyimino)acetamido]-3-ethyl-aminomethylceph-3-em-4-carboxylic acid (more polar isomer),having the following n.m.r. in solvent A: 1.19(t, 3H); 1.43(d, 3H);2.94(q, 2H); 3.37(d, 1H); 3.41(d, 1H); 3.6(d, 1H); 3.76(d, 1H);4.61(q, 1H); 5.03(d, 1H); 5.75(d, 1H); 6.78(s, 1H).Retention time 3.25 minutes on a reversed phase Partisil PXS10/25 ODS-2 column, using MeOH/water/HOAc 25:75:1 aseluant.
EXAMPLE 220
Dry dichloromethane (5 ml) was cooled to -10.degree. and oxalyl chloride (1.15 mM), followed by DMF (1.15 mM), was added, and the mixture was stirred at -15.degree. to -10.degree. for 1 hour. To this was then added 1-(Z)-chloroethylene-2-(2-formamidothiazol-4-yl)acetic acid (1 mM), followed by N-methylmorpholine (1.15 mM), and the mixture was stirred for 1 hour at -15.degree. to -10.degree..
In another flask a solution of 7-amino-3-(1-methyl-4-pyridinio)aminomethylceph-3-em-4-carboxylic acid (1.15 mM), as its partial salt with 1.76 moles TFA, in dimethylacetamide (6 ml) was treated with N-methylmorpholine (2.08 mM), and the pink suspension was syringed into the flask containing the activated acid, is followed by N-methylmorpholine (1.15 mM). After stirring for 1 hour, at -10.degree. and 1 hour at ambient temperature, the mixture was filtered. The solution was evaporated to dryness, and deformylated by solution in methanol (20 ml) and treating with concentrated hydrochloric acid (0.5 ml), at ambient temperature for 2 hours. After removal of the methanol, the residue was dissolved in water, the pH was adjusted to 5.2 with aqueous NaHCO.sub.3, and the product was purified by chromatography on Diaion CHP20P resin to give 7-[2-(2-aminothiazol-4-yl)-2-(Z)-chloromethyleneacetamido]-3-(1-methyl-4-pyridinio)aminomethylceph-3-em-4-carboxylic acid, in 5% yield, having the following n.m.r. in solvent A: 3.27 (d, 1H); 3.5 (d, 1H); 3.84 (s, 3H); 4.19 (d, 1H); 4.41 (d, 1H); 5.07 (d, 1H); 5.66 (d, 1H); 6.38 (s, 1H); 6.82 (s, 1H); 6.89 (d, 1H); 7.26 (d, 1H); 8.01 (d, 1H); 8.18 (d, 1H).
EXAMPLES 221-228
To a stirred suspension of 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid (240 mg, 0.5 mmole) and triethylamine (200 ul, 1.4 mmole) in EtOH (10 ml) at 25.degree. was added 1-methyl-4-methylthioquinazolinium iodide (160 mg. 0.5 mmole). After 1.5 hours the solution was evaporated to dryness under reduced pressure, the residue dissolved in water (10 ml), the solution acidified with excess 5% v/v aqueous HOAc and the insoluble material filtered off. The filtrate was applied to a Dianion HP20 column and the product purified by gradient elution with MeOH. Using this general process and the appropriate quaternary heterocycle, the following compounds were obtained:
__________________________________________________________________________ ##STR270##Example YieldNo. R.sub.1 R.sub.2 R.sub.3 R.sub.4 % Footnotes__________________________________________________________________________221 CH.sub.3 CH.sub.3 CH.sub.3 H 11 1, 2, 3222 CH.sub.3 H H H 9 4, 5, 6223 CH.sub.2 CHCH.sub.2 H H H 44 7, 8, 6224 ##STR271## H H H 33 9, 10, 6225 CH.sub.3 H H CH.sub.2 CH.sub.3 29 11, 12, 13, 5, 6226 CH.sub.2 CHCH.sub.2 H H CH.sub.2 CH.sub.3 22 14, 15, 13, 8, 6227 CH.sub.2 CHCH.sub.2 H H ##STR272## 46 16, 17, 18, 19, 8, 6228 CH.sub.3 H H ##STR273## 35 20, 21, 22, 5, 6__________________________________________________________________________Footnotes1. n.m.r. in solvent A: 1.41(s, 3H); 1.43(s, 3H); 2.52(m, 6H); 3.49(d,1H); 3.72(d, 1H); 3.94(s,3H); 4.26(d, 1H); 4.82(d, 1H); 5.02(d, 1H); 5.76(d, 1H); 6.75(s, 1H);8.74(s, 1H).2. The 1,5,6-trimethyl-4-methylthiothieno[2,3-d]pyrimidinium iodide maybe prepared asfollows: A solution of 5,6-dimethyl-4-methylthiothieno[2,3-d]pyrimidinein CH.sub.2 Cl.sub.2 wastreated with trimethyloxonium tetrafluoroborate and stirred, under argon,at room tempera-ture overnight. The solution was evaporated to dryness to give a whitesolid. N.mr. in d.sub.6DMSO: 2.6(s, 6H); 2.8(s, 3H); 4.2(s, 3H); 9.4(s, 1H).3. 5,6-Dimethyl-4-methylthiothieno[2,3-d]-pyrimidine may be prepared asfollows: A solutionof 5,6-dimethyl-1H-thieno[2,3-d]pyrimidin-4-one in CH.sub.3 CN wastreated, at reflux for 3 hr,with Lawesson's reagent. The reaction mixture was poured into water andextracted withethyl acetate. The organic extracts were combined and extracted severaltimes with 2NNaOH. The aqueous extracts were acidified with HCl and re-extracted withethyl acetate,the extracts washed with water, dried and evaporated to dryness. Thecrude 5,6-dimethyl-1H-thieno[2,3-d]pyrimidin-4-thione so obtained (M.sup.+ = 196) wasalkylated withmethyl iodide in aqueous sodium hydroxide to give, after chromatographyon silica, 5,6-dimethyl-4-methylthiothieno[2,3-d]pyrimidine. n.m.r. in CDCl.sub.3 :2.5(s, 6H); 2.6(s,3H); 8.7(s, 1H). This product was quaternised by heating with excessmethyl iodide underreflux for 18 hours.4. n.m.r. in solvent A: 1.4(s, 3H); 1.44(s, 3H); 3.46(d, 1H); 3.62(d,1H); 4.02(s, 3H); 4.55(d,1H); 4.9(d, 1H); 5.02(d, 1H); 5.82(d, 1H); 6.72(s, 1H); 7.90(s, 1H);8.85(s, 1H).5. The 1-methyl-4-methylthiothieno[2,3-d]pyrimidinium tetrafluoroboratewas prepared from 4-methylthiothieno[2,3-d]pyrimidine and trimethyloxonium tetrafluoroborateby a procedureanalogous to that described in footnote 2. n.m.r. in d.sub.6 DMSO: 2.9(s,3H); 4.34(s, 3H); 7.88(d,1H); 8.28(d, 1H); 9.52(s, 1H).6. The 4-methylthiothieno[2,3-d]pyrimidine was prepared from1H-thieno[2,3-d]pyrimidine-4-oneby a two-stage procedure analogous to that described in footnote 3.n.m.r. in CDCL.sub.3 : 2.72(s, 3H); 7.35(d, 1H); 7.45(d, 1H); 8.85(s, 1H).7. n.m.r. in solvent A: 1.36(s, 3H); 1.38(s, 3H); 3.36(d, 1H); 3.6(d,1H); 4.56(d, 1H); 4.88(d,1H); 5.04(d, 3H); 5.42(d, 1H); 5.48(d, 1H); 5.75(d, 1H); 6.04(m, 1H);6.75(s, 1H); 7.86(d,1H); 7.93(d, 1H); 8.92(s, 1H).8. The 1-allyl-4-methylthiothieno[2,3-d]pyrimidinium bromide was preparedfrom 4-methylthio-thieno[2,3-d]pyrimidine and allyl bromide in refluxing acetonitrile. Theexcess allyl bromideand solvent were removed by evaporation. n.m.r. in d.sub.6 DMSO: 2.9(s,3H); 5.45(d, 2H);5.6(d, 1H); 5.75(d, 1H); 6.18(m, 1H); 7.88(d, 1H); 8.32(d, 1H); 9.72(s,1H).9. n.m.r. in solvent A: 1.42(s, 3H); 1.44(s, 3H); 3.40(d, 1H); 3.62(d,1H); 4.6(d, 1H); 4.92(d,1H); 5.08(d, 1H); 5.76(d, 1H); 5.84(s, 2H); 6.74(s, 1H); 7.75(d, 1H);7.8(d, 1H); 7.92(d, 1H);8.24(d, 1H); 9.18(s, 1H).10. The 4-methylthio-1-p-nitrobenzylthieno[2,3-d]-pyrimidinium bromidewas prepared asfollows: 4-Methyl-thiothieno[2,3-d]pyrimidine in CH.sub.3 CN was treatedwith p-nitrobenzylbromide and the solution refluxed for 8 hr. The solution was evaporatedto dryness and theresidue triturated with CH.sub.2 Cl.sub.2. The insoluble solid wasfiltered off, anddried. n.m.r. in d.sub.6 DMSO: 2.94(s, 3H); 6.16(s, 2H); 7.82(dd, 2H);8.2(d, 1H); 8.25(d, 1H);9.8(s, 1H).11. To a stirred suspension of 3-ethylaminoethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.-- Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid (260 mg. 0.5 mM)in DMF(5 ml) was added Et.sub.3 N (150 .mu.l, 1.1 mM) followed by1-methyl-4-methylthiothieno[2,3-d]-pyrimidinium tetrafluoroborate (150 mg, 0.53 mM). The mixture was stirredat room temper-ature overnight before being poured into H.sub.2 O (100 ml). The pH ofthe solution was adjustedto 3.5 with glacial acetic acid and the solution applied to a DiaionHP20SS column. Theproduct was eluted using H.sub.2 O and CH.sub.3 CN and the appropriatefractions were freeze-dried.12. n.m.r. in solvent A: 1.35(br t, 3H); 1.43(s, 3H); 1.44(s, 3H);3.38(d, 1H); 3.54(d, 1H); 4.06(m, 2H); 4.06(s, 3H); 4.79(d, 1H); 5.16(d, 1H); 5.28(d, 1H); 5.85(d, 1H);6.71(s, 1H); 7.82(d,1H); 7.95(d, 1H); 8.85(s, 1H).13. The 3-ethylaminomethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.--Z)-1-carboxy-1-methylethoxyimino)-acetamido]ceph-3-em-4-carboxylic acid was prepared as follows:3-Aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.-- Z)-1-carboxy-1-methylethoxyimino)acetamido]cep-3-em-4-carboxylic acid (2 g, 4 mM) was suspended in MeOH (200 ml), andEt.sub.3 N (570 .mu.l,4.1 mM) followed by sodium cyanoborohydride were added. The mixture wasstirred,under argon, at room temperature and treated, dropwise, during 40 min.with a solutionof acetaldehyde (276 .mu.l, 4.9 mM) in methanol (2 ml). After 3 hr. themethanol wasevaporated under reduced pressure and the product was purified bychromatography onDiaion HP20SS using H.sub.2 O and CH.sub.3 CN in a graded elution. Theappropriatefractions were combined and freeze dried. N.m.r. in solvent A: 1.15(t,3H); 1.44(s, 3H);1.46(s, 3H); 2.82(q, 2H); 3.36(d, 1H); 3.45(d, 1H); 3.59(d, 1H); 3.76(d,1H); 5.0(d, 1H);5.77(d, 1H); 6.74(s, 1H).14. The procedure described in Footnote 11 was followed, but startingwith 1-allyl-4-methylthiothien-[2,3-d]pyrimidinium bromide.15. n.m.r. in solvent A: 1.32(t, 3H); 1.45(br s, 6H), 3.24(d, 1H);3.46(d, 1H); 4.10(m,2H); 4.91(d, 1H); 5.08(d, 1H); 5.26(d, 1H); 5.45(d, 1H); 5.52(d, 1H);5.72(d, 1H);6.05(m, 1H); 6.7(s, 1H); 7.79(d, 1H); 7.92(d, 1H); 8.89(s, 1H).16. The procedure described in Footnote 11 was followed, but startingwith 3-(2-fluoro-1 .sub.-- H-imidazol-4-ylmethyl)aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.--Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid and1-allyl-4-methylthio-thieno[2,3-d]pyrimidinium bromide.17. n.m.r. in solvent A: 1.42(s, 3H); 1.46(s, 3H); 3.12(d, 1H); 3.37(d,1H); 4.93(d, 1H);4.97(d, 1H); 5.0-5.1(m, 4H); 5.18(d, 1H); 5.44(d, 1H); 5.5(d, 1H);5.72(d, 1H); 6.02(m, 1H); 6.72(s, 1H); 6.82(s, 1H); 7.79(d, 1H); 8.04(d, 1H); 8.87(s,1H).18. 3-(2-Fluoro-1H-imidazol-4-ylmethyl)aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.-- Z)-1-carboxy-1-methylethoxyimino)-acetamido]ceph-3-em-4-carboxylic acid wasprepared from3-(2-fluoro-1-triphenylmethylimidazol-4-ylmethyl)aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.-- Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid by treatment inCH.sub.2 Cl.sub.2 at room temperature with p-toluene sulphonic acidH.sub.2 O. Thereaction mixture was poured into water and the aqueous layer applied to aDiaion HP20SScolumn. The product was obtained by gradient elution with CH.sub.3CN/H.sub.2 O and freeze-drying the appropriate fractions. N.m.r. in solvent A: 1.45(br s, 6H);3.42(d, 1H); 3.54(d,1H); 3.68(d, 1H); 3.83(d, 1H); 3.92(s, 2H); 5.03(d, 1H); 5.75(d, 1H);6.76(s, 1H); 6.93(s, 1H).19. The 3-(2-fluoro-1-triphenylmethylimidazol-4-ylmethyl)aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.-- Z)-1-carboxy-1-methylethoxyimino)-acetamido]ceph-3-em-4-carboxylicacid was prepared from 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((.sub.-- Z)-1-carboxy-1-methylethoxy imino)acetamido]ceph-3-em-4-carboxylic acid and2-fluoro-4-formyl-1-triphenyl-methylimidazole by a procedure analogous to that described in footnoteN.m.r. in solvent A: 1.44(s, 3H); 1.46(s, 3H); 3.32(d, 1H); 3.4(d, 1H);3.58(d, 1H); 3.75(d,1H); 3.92(s, 2H); 5.0(d, 1H); 5.71(d, 1H); 6.75(s, 1H); 6.8(s, 1H);7.13(d, 6H); 7.43(m, 9H).20. The procedure described in Footnote 11 was followed but starting with3-(pyrid-3-ylmethyl)aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-(( .sub.-- Z)-1-carboxy-1-methylethoxyimino)acetamido]-ceph-3-em-4-carboxylic acid.21. N.m.r. in solvent A: 1.42(s, 6H); 3.35(br d, 1H); 3.54(br d, 1H);4.04(s, 3H); 5.02(d, 1H);5.05-5.55(m, 4H); 5.74(d, 1H); 6.72(s, 1H); 7.35(m, 1H); 7.79(br d, 1H);7.92(m, 2H); 8.48(br s, 1H); 8.93(s, 1H).22. 3-(Pyrid-3-ylmethyl)aminomethyl-7-[2-(2-aminothiazol-4-yl)-2((.sub.-- Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid was preparedfollowing theprocedure described in Footnote 3, but with 3-formylpyridine as thealdehyde. N.m.r. insolvent A: 1.4(s, 3H); 1.44(s, 3H); 3.39(d, 1H); 3.48(d, 1H); 3.65(d,1H); 3.73(d, 1H); 4.12(s,2H); 5.02(d, 1H); 5.76(d, 1H); 6.76(s, 1H); 7.38(m, 1H); 7.87(d, 1H);8.54(br s, 1H); 8.62(s,1H).
EXAMPLES 229-233
To a stirred suspension of the appropriate 3-aminomethylceph-3-em-4-carboxylic acid (0.82 mmole) and triethylamine (1.8 mmole) in DMF (10 ml) at 25.degree. was added the appropriate 6-chloro-1,2,4-triazolo[4,3-b]-pyridazinium iodide (0.84 mmole). After stirring overnight, the mixture was poured into water (100 ml), the pH adjusted to 3.5 with glacial acetic acid and the solution filtered. The filtrate was applied to a Diaion HP20P resin column, and the product was obtained by gradient elution using water and acetonitrile. The appropriate fractions were freeze-dried.
__________________________________________________________________________ ##STR274##Example YieldNo. R.sub.1 R.sub.2 R.sub.3 % Footnotes__________________________________________________________________________229 2-CH.sub.3 H H 8 1, 2230 1-CH.sub.3 H H 21 3, 4231 1-CH.sub.3 Cl H 16 5, 6232 1-CH.sub.3 H CH.sub.2 CH.sub.3 12 7, 8, 4233 9, 10, 11 ##STR275##__________________________________________________________________________Footnotes1. n.m.r. in solvent A: 1.46(br s, 6H); 3.42(d, 1H); 3.65(d, 1H);4.0-4.2(br s, 4H);4.3-4.5(br s, 1H); 5.04(d, 1H); 5.75(d, 1H); 6.74(s, 1H); 7.3(d, 1H);8.04(d, 1H);approximately 11.0(s, 1H).2. The triazolopyridazinium salt may be prepared as follows:6-chloro-1,2,4-triazolo-[4,3-b]pyridazine in methanol was treated with methyl iodide, and thesolution was refluxed for 2 hr. Anhydrous ether was then added to thereaction mixtureand the precipitated solid so obtained was filtered off, washed with alittle ether anddried under vacuum, m. pt. 238-40.degree..3. n.m.r. in solvent A: 1.42(s, 3H); 1.45(s, 3H); 3.38(d, 1H); 3.64(d,1H); 4.1(br s,4H); 4.38(m, 1H); 5.02(d, 1H); 5.73(d, 1H); 6.74(s, 1H); 7.53(bs, 1H);8.35(d, 1H);9.65(s, 1H).4. The 6-chloro-1-methyl-1,2,4-triazolo[4,3-b]pyridazinium iodide may bepreparedas follows: 6-chloro-1,2,4-triazolo[4,3-b]pyridazine and methyl iodidewere refluxedin acetonitrile for 4 hr. The reaction mixture was cooled and the solidwhich hadprecipitated during the reaction was filtered off, washed with a littleacetonitrile, anddried, m pt 208-210.degree..5. n.m.r. in solvent A: 1.42(s, 3H); 1.44(s, 3H); 3.45(d, 1H); 3.66(d,1H); 4.1(m, 4H);4.5(d, 1H); 5.04(d, 1H); 5.78(d, 1H); 6.74(s, 1H); 7.57(m, 1H); 8.34(d,1H).6. The 3,6-dichloro-1-methyl-1,2,4-triazolo[4,3-b]pyridazinium iodide waspreparedfrom 3,6-dichloro-1,2,4-triazolo[4,3-b]pyridazine and methyl iodide inacetonitrile.N.m.r. in d.sub.6 DMSO: 4.38(s, 3H); 8.45(d, 1H); 9.12(d, 1H).7. Prepared from 3-ethylaminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((.sub.-- Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid.8. n.m.r. in solvent A: 1.14(t, 3H); 1.42(s, 6H); 3.2(d, 1H); 3.46(d,1H); 3.6(m, 1H);3.75(m, 1H); 4.18(s, 3H); 4.64(m, 2H); 5.06(d, 1H); 5.7(d, 1H); 6.72(s,1H); 8.2(br s,1H); 8.52(d, 1H); 9.69(s, 1H).9. Sodium hydrogen carbonate was used in place of triethylamine. Yield3%.10. n.m.r. in solvent A: 1.42(s, 3H); 1.45(s, 3H); 3.42(d, 1H); 3.65(d,1H); 4.07(s, 3H);4.24(d, 1H); 4.46(d, 1H); 5.01(d, 1H); 5.72(d, 1H); 6.74(s, 1H); 7.82(d,1H); 7.87(d,1H); 8.64(dd, 1H); 8.91(dd, 1H).11. The 3-chloro-1-methyl-1,2,4-triazolo[4,3-b]pyradizinium iodide waspreparedfrom 3-chloro-1,2,4-triazolo[4,3-b]pyridazine and methyl iodide inacetonitrile.The product which separated from solution was filtered off washed with alittleCH.sub.3 CN and ether and dried, m. pt. 225-227.degree..
EXAMPLE 234
The process described in Example 221 was repeated using the appropriate 3-ethylaminomethyl cephem and quaternary heterocycle to give the following compound:
__________________________________________________________________________ ##STR276##Example No. R Yield % Footnotes__________________________________________________________________________234 ##STR277## 44 1, 2, 3__________________________________________________________________________Footnotes1. n.m.r. in solvent A: 1.34(m, 3H); 1.42(br s, 6H); 2.08(s, 3H); 3.38(d,1H);3.53(d, 1H); 4.02(m, 2H); 4.80(d, 1H); 5.14(d, 1H); 5.2-5.45(overlappings, d, 3H); 5.68(s, 1H); 5.84(d, 1H); 6.7(s, 1H); 7.79(d, 1H); 7.9(d, 1H);8.84(s, 1H).2. The chemical shifts quoted probably refer to the major E isomer, butthere is alsoevidence of the minor Z in the n.m.r. spectrum.3. The starting material was prepared from 4-methylthiothieno[2,3-d]pyrimidine and2-bromomethyl-1-cyanoprop-1-ene (mixture of E and Z isomers in the ratio9:1)in CH.sub.3 CN. The precipitate was filtered off and washed with CH.sub.3CN and dried.N.m.r. in d.sub.6 DSMO: 2.1(s, 3H); 2.92(s, 3H); 5.64(s, 2H); 5.86(s,1H); 7.86(d, 1H);8.24(d, 1H); 9.6(s, 1H).
EXAMPLE 235 ##STR278##
3-Aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid (242mg, 0.5 mmole) was suspended in DMF (3 ml) and water (1 ml) and treated with 2-methylthio-1,2,4-thiadiazolo[2,3-a]pyridinium bromide (132 mg, 0.5 mmole) and then with sodium hydrogen carbonate (240 mg, 2.5 mmole) suspended in water (0.5 ml). The solution was stirred at room temperature overnight before being diluted with water (25 ml) and applied to a Diaion HP20SS column. Following gradient elution with acetonitrile/water the product was obtained by freeze-drying the appropriate fractions. Yield 11%. N.m.r. in solvent A: 1.45 (s, 3H); 1.46 (s, 3H), 3.64 (d, 1H); 3.76 (d, 1H); 4.34 (d, 1H); 4.88 (d, 1H); 5.07 (d, 1H); 5.78 (d, 1H); 6.73 (s, 1H); 7.0 (m, 1H); 7.13 (d, 1H); 7.72 (m, 1H); 8.17 (dd, 1H).
EXAMPLES 236-243
A solution of 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamidolceph-3-em-4-carboxylic acid (0.5 mm) and NaHCO.sub.3 (2.5 mM) in water (5 ml) was treated with an appropriate chloro- or methylsulphinyl-substituted quaternary heterocycle, at room temperature for 18 hrs. The solution was acidified with HOAC and sufficient sodium acetate added to redissolve any precipitate. The product was isolated by chromatography on an HP20 gravity column, eluting with CH.sub.3 CN/H.sub.2 O mixtures and then freeze-drying the appropriate fractions. The following compounds were obtained:
______________________________________ ##STR279##Example YieldNo. R % Footnotes______________________________________236 ##STR280## 34 1, 2237 ##STR281## 36 3, 4238 ##STR282## 27 5, 6239 ##STR283## 26 7, 8240 ##STR284## 33 9, 10241 ##STR285## 57 11, 12242 ##STR286## 48 13, 14243 ##STR287## 16 15, 16______________________________________Footnotes1. n.m.r. in solvent A: 1.45(2xs, 6H); 3.4(q, 2H); 4.3(q. 2H);5.05(d, 1H); 5.45(s, 2H); 5.7(d, 1H); 6.7(s, 1H); 6.9-8.5(6xm, 8H).2. The starting pyridinium salt was obtained by adding 4-hydroxy-pyridine (10 mM) to a stirred suspension of hexane-washed sodiumhydride (10 mM) in DMF (15 ml). To this solution was added asolution of 4-chloromethylpyridine (10 M) in ether (5 ml), (ob-tained by treating the corresponding 4-chloromethylpyridinehydrochloride with NaHCO.sub.3 in aqueous solution, extractingwith ether and drying), and the mixture was stirred at roomtemperature for 18 hrs. The solvents were evaporated, and theresidue was purified by MPLC, on Merck silica gel 9385, elutingwith MeOH/CH.sub.2 Cl.sub.2, 8-10:92-90, to give 1-(2-pyridylmethyl)-pyrid-4-one. Yield 65%. N.m.r. in CDCl.sub.3 : 5.0(s, 2H); 7.2-7.8(m,4H); 6.35(d, 2H); 8.6 (d, 2H). This pyridone (1.5 mM) and tosylchloride (1.65 mM) were heated in DMF at 80.degree. for 45 minutes,the solvent was evaporated, and the residue triturated withCH.sub.2 Cl.sub.2 to give 4-chloro-1-(2-pyridylmethyl)pyridiniumtosylatein 70% yield. N.m.r. in d.sub.6 DMSO: 2.3(s, 3H); 6.0(s, 2H); 7.1(d,2H); 7.5(d, 2H); 7.4-8.6(m, 6H); 9.15(d, 2H).3. N.m.r. in solvent A: 1.45(2xs, 6H); 3.4(q, 2H), 4.3(q, 2H); 5.1(d,1H); 5.35(s, 2H); 5.8(d, 1H); 6.75(s, 1H); 6.95(q, 1H); 7.1(q, 1H);7.4(d, 2H); 8.2(d, 1H); 8.3(d, 2H); 8.4(d, 1H).4. The starting pyridinium salt was obtained as follows: A solutionof 4-chloromethyl-1-oxidopyridinium chloride (5 mM) in ether wasbasified with NaHCO.sub.3 solution. The ether solution was separatedand dried, 4-methylthiopyridine was added, the solvent wasevaporated and the residue was kept at room temperature for 18hrs. Trituration gave 4-methylthio-1-(1-oxido-pyrid-4-ylmethyl)py-ridinium chloride. N.m.r. in d.sub.6 DMSO: 2.7(s, 3H); 5.75(s, 2H);7.6(d, 2H); 8.0(d, 2H); 8.25(d, 2H); 9.05(d, 2H). This thiomethylcompound (2 mM) was suspended in CH.sub.2 Cl.sub.2 (10 ml) at 0.degree.,andsufficient TFA was added to give a solution. A solution ofMCPBA (2 mM) in CH.sub.2 Cl.sub.2 (3 ml) was added, and the solutionwas allowed to warm to room temperature, then stirred for 1 hr.The solvents were evaporated, and the residue was trituratedseveral times with ether, to remove excess MCPBA, to give 4-methylsulphinyl-1-(1-oxidopyrid-4-ylmethyl)-pyridinium chloride,which was used without further purification or characterisation.5. N.m.r. in solvent A: 1.4(2xs, 6H); 3.35(q, 2H), 4.3(q, 2H);5.05(d, 1H); 5.45(s, 2H); 5.7(d, 1H); 6.7(s, 1H); 6.5-8.4(6xm, 8H).6. The starting pyridinium salt was prepared by the process ofFootnote 4, except that the reactants were warmed together toform a homogeneous melt, then heated at 100.degree. for 15 minutes,cooled and triturated with CH.sub.2 Cl.sub.2. N.m.r. in d.sub.6 DMSO:2.7(s,3H); 5.9(s, 2H); 7.5, 7.95 and 8.35(m, 6H); 8.95(d, 2H). Themethylthio compound was then oxidised to the required methyl-sulphinyl compound as described in Footnote 4.7. Reaction solvent was H.sub.2 O/CH.sub.2 CN, (4:1), N.m.r. in solventA:1.45(2xs, 6H); 3.4(q, 2H), 4.3(q, 2H); 5.05(d, 1H); 5.4(s, 2H); 5.7(d,1H); 6.7(s, 1H), 6.95(q, 1H); 7.4(q, 1H); 7.55(s, 2H); 8.15(q, 1H);8.35(q, 1H).8. The starting pyridinium salt was prepared by the process ofFootnote 4, from 4-bromomethyl-2,6-dichloropyridine, except thatthis starting material reacted exothermically with 4-methylthio-pyridine, so the reaction mixture was kept below 40.degree. until thereaction was largely complete, then heated at 100.degree. for 5 minutes.The product was crystallised form HCl/EtOH. N.m.r. in d.sub.6DMSO: 2.75(t, 3H); 5.95(s, 2H); 7.75(s, 2H); 7.95(d, 2H); 9.05(d,2H). The methylthio compound was oxidised to the requiredmethylsulphinyl compound as in Footnote 4. N.m.r. in d.sub.6 -DMSO/CDCl.sub.3 : 2.95(s, 3H); 5.95(s, 2H); 7.7(s, 2H); 8.4(d, 2H); 9.3(d,2H).9. N.m.r. in solvent A: 1.4(2xs, 6H); 2.25(s, 3H); 3.4(q, 2H); 4.3(q,2H); 5.05(d, 1H); 5.3(s, 2H); 5.7(d, 1H); 6.7(s, H); 6.9(d, 1H);7.25(d), 8.05(d, 1H); 8.8(d, 1H).10. The starting pyridinium salt was obtained as follows:5-Hydroxymethyl-6-methyluracil (15 mM) was suspended inglacial acetic acid (10 ml) and the mixture was saturated with HClgas, then heated under reflux for 1/2 hour until the solid materialwas dissolved, and filtered. On cooling, 5-chloromethyl-6-methyl-uracil was obtained, n.m.r. in d.sub.6 DMSO: 2.15(s, 3H); 4.45(s, 2H).This compound was treated with a solution of 4-methylthio-pyridine in CH.sub.2 Cl.sub.2, the solvent was evaporated, and theresiduewas heated at 100.degree. for 10 minutes. It stayed solid throughout,butformed the required quaternary salt. Yield 93%. N.m.r. in d.sub.6DMSO: 2.25(s, 3H); 2.65(s, 3H); 5.3(s, 2H); 7.8(d, 2H); 8.7(d, 2H).This material was oxidised to the required 4-methylsulphinyl start-ing material, as in Footnote 4.11. N.m.r. in solvent A: 1.45(2xs, 6H); 2.1(s, 3H); 2.45(s, 3H);3.4(q, 2H); 4.3(q, 2H); 5.0(d, 1H); 5.2(s, 2H); 5.7(d, 1H); 6.75(s,1H); 6.9(q, 1H); 7.3(q, 1H); 8.0(q, 1H); 8.25(q, 1H).12. The starting pyridinium salt was obtained by the process ofFootnote 4. N.m.r. of 4-methylthiopyridinium salt, in d.sub.6 DMSO:2.15(s, 3H); 2.5(s), 2.7(s, 3H); 5.75(s, 2H); 7.95(d, 2H); 8.85(d, 2H).The compound was oxidised to the required 4-methylsulphinylstarting material as in Footnote 4.13. n.m.r. in solvent A: 0.8-1.0(m, 4H); 1.45(2xs, 6H); 1.9(partiallyobscured by solvent peaks); 3.4(q, 2H); 4.3(q, 2H); 5.0(s, 2H);5.05(d, 1H); 5.7(d, 1H); 6.0(s, 1H); 6.7(s, 1H); 6.9(d, 1H); 7.15(d,1H); 8.0(d, 1H); 8.15(d, 1H).14. The starting pyridinium salt was obtained as follows: 6-Chloro-methyl-2-cyclopropyl-4-hydroxypyridine (10 mM) and 4-methyl-thiopyridine (10 mM) were thoroughly mixed and heated at 100.degree.for 1/2 hour. The mixture partly melted, then re-solidified. Oncooling, the product was triturated with CH.sub.2 Cl.sub.2 to give the4-methylthiopyridinium salt. Yield 73%. N.m.r. in d.sub.6 DMSO0.6-1.1(m, 4H); 2.0(m, 1H); 2.7(s, 3H); 5.55(s, 2H); 6.2(s, 1H);7.95(d, 2H); 8.7(d, 2H). This product was oxidised to the required4-methylsulphinyl pyridinium salt, as in Footnote 4.15. N.m.r. in solvent A: 1.4(s, 6H); 3.1(s, 3H); 3.4(q, 2H); 4.3(q,2H); 5.05(d, 1H); 5.3(s, 2H); 5.75(d, 1H); 6.75(s, 1H); 7.15(d, 1H);7.7(d, 1H); 6.9-8.45(m, 5H).16. The starting pyridinium salt, in this case the tetrafluoroborate,was obtained by reacting 5-bromomethyl-1-methylisatin with 4-methylthiopyridine to give 1-(1-methyl-5-isatinylmethyl)-4-methylthiopyridinium bromide, m.p. 227.5-228.5 (crystallised fromEtOH). This salt was dissolved in MeOH/H.sub.2 O, 1:1 v/v (30 ml)and the solution was stirred while silver tetrafluoroborate (1 mM)in water (5 ml) was added. The yellow silver bromide was filteredoff through kieselguhr, the filter cake was suspended in water (50ml) and again filtered, and the filtrates were combined andevaporated to give the required tetrafluoroborate salt.
EXAMPLE 244-245
7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]-3-ethylaminomethylceph-3-em-4-carboxylic acid (205 mg) was dissolved in a mixture of sodium bicarbonate (336 mg) in water (5 ml), and acetonitrile (5 ml). 4-Chloro-1-(4-nitrobenzyl)pyridinium chloride (168 mg) was added, stirred for 10 minutes to achieve solution, then stood at room temperature overnight. The solvent was evaporated, and the residue was purified by running on an RP18 column (in sodium salt form), eluting with H.sub.2 O/CH.sub.3 CN, 100-70:0-30, to give 7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]-3-[N-(1-p-nitrobenzyl-4-pyridinio)-N-ethyl]aminomethylceph-3-em-4-carboxylic acid. Yield 27.5%. N.m.r. in solvent A: 1.1 (t, 3H), 1.45 (2xs, 6H); 3.2-3.7 (m, 4H); 4.5-4.7 (q, 2H); 5.15 (d, 1H); 5.55 (s, 2 H); 5.85 (d, 1H); 6. 7 (s, 1H); 7.1-7.4 (m, 2H); 7.7 (d, 2H); 8.3 (d, 2H); 8.45 (d, 2H).
In a similar manner, using the corresponding N-carboxymethyl cephem, there was obtained 7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)-acetamido]-3-[N-carboxymethyl-N-(1-p-nitrobenzyl -4-pyridinio]aminomethylceph-3-em-4-carboxylic acid. Yield 19%.
N.m.r. in solvent A: 1.4 (2xs, 6H); 3.25-3.6 (q, 2H); 4.4 (s, 2H); 4.6 (s, 2H); 5.15 (d, 1H); 5.55 (s, 2H); 5.85 (d, 1H); 6.7 (s, 1H); 7.05-7.2 (m, 1H); 7.4-7.55 (m, 1H); 7.6-7.7 (d, 2H); 8.2-8.3 (d, 2H); 8.4-8.6 (m, 2H).
EXAMPLE 246
The process of Example 244 was repeated, using the appropriate N-substituted cephem starting material, to give the following compound: ##STR288##
N.m.r. in solvent A: 1.15 (t, 3H); 1.7-1.9 (m, 2H); 2.2-2.5 (m, 4H); 3.0-3.5 (q, 2H); 3.5-3.8 (m, 2H); 4.4-4.8 (q, 2H); 5.05 (d, 1H); 5.5 (s, 2H); 5.7 (d, 1H); 6.7 (s, 1H); 7.05-7.25 (m, 1H); 7.6-7.7 (m, 3H); 8.2-8.3 (d, 2H); 8.35-8.4 (d, 2H).
EXAMPLE 247
3-Acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxycyclobutyloxyimino)acetamido]ceph-3-em-4-carboxylic acid (460 mg) was suspended in water (20 ml) with sufficient NaHCO.sub.3 to obtain a solution. 5-Amino-2-methylisoquinolinium iodide (244 mg) was added, and the solution was heated under argon for 3hrs. at 85.degree., maintaining the pH at 7.5 by small additions of HOAC or aqueous NaHCO3. The solution was cooled, made to pH5 with HOAC, and run onto a column of HP20SS resin. The column was eluted with H.sub.2 O/CH.sub.3 CN, 100-80:0-20. The fractions containing the product, 7-[2-(2-aminothiazol- 4-yl)-2-((Z)-1-carboxycyclobutyloxyimino)acetamido]-3-[(2-methyl-5-isoquinolinio)aminomethyl]ceph-3-em-4-carboxylic acid, were combined and lyophilised. N.m.r. in solvent A: 1.85 (m, 2H); 2.35 (m, 4H); 3.45 (m, 2H); 4.4 (m, 2H); 4.4 (s, 3H); 5.05 (d, 1H); 5.75 (d, 1H); 6.75 (s, 1H); 7.3 (d, 1H); 7.55 (d, 1H); 7.75 (t, 1H); 8.54 (d, 1H); 8.65 (d, 1H), 9.7 (s, 1H).
EXAMPLE 248
The process described in Example 221 was repeated, using 7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]-3-(3-pyridylmethylaminomethyl)ceph-3-em-4-carboxylic acid and 1-allyl-4-chloropyridinium bromide as starting materials, to give 3-[N-(1-allyl-4-pyridinio)-N-(3-pyridylmethyl)aminomethyl]-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-methylethoxyimino)acetamido]ceph-3-em-4-carboxylic acid. N.m.r. in solvent A: 1.65 (s, 3H), 1.7 (s, 3H); 3.35 (d, 1H); 3.7 (d, 1H); 4.7-5.3 (m, 7H), 5.4 (d, 1H); 5.5 (d, 1H); 5.95 (d, 1H); 6.1 (m, 1H); 7.0 (s, 1H); 7.1-7.5 (br m, 2H); 7.65 (t, 1H); 8.0 (d, 1H); 8.3 (m, 2H); 8.7 (s, 2H).
The cephem starting material was obtained as in Example 62, Footnote 14.
1-Allyl-4-chloropyridinium bromide was obtained by quaternising 4-chloropyridine with allyl bromide.
EXAMPLES 249-254
The process described in Example 54 was repeated, to obtain the following compounds:
__________________________________________________________________________ ##STR289##Example No. R R.sup.1 Yield % Footnotes__________________________________________________________________________249 Et ##STR290## 6 1, 2250 Et ##STR291## 22 3251 Et ##STR292## 26 4252 (CH.sub.2).sub.2 CN ##STR293## 20 5253 H ##STR294## 9 6, 7254 (CH.sub.2).sub.2 CN ##STR295## 9 8__________________________________________________________________________Footnotes1. Initially, 1 eq. of quaternary heterocyclic salt and NaHCO.sub.3reacted with theCAZAMCA derivative, and further portions of salt and NaHCO.sub.3 weresub-sequently added up to a total of 5 eq. and 10 eq. respectively.2. Eluant portion 20-35:80-65:1. N.m.r. in solvent B: 1.85(s, 3H);1.55(s, 6H); 3.5(m,2H); 3.7(m, 2H); 4.55(d, 1H); 5.15(d, 1H); 5.2(d, 1H); 5.8(d, 1H);7.05(s, 1H); 7.45(d,1H); 7.8(d, 1H); 8.2(d, 1H); 9.0(d, 1H).3. Reaction was carried out in DMF, using 2 eq. of Et.sub.3 N in place ofNaHCO.sub.3.Eluant portion 25-30:75-70:1. N.m.r. in solvent B: 1.2(t, 3H); 2.15(s,6H); 3.4-3.8(m,4H); 4.5-4.8(m, 2H); 5.1(d, 1H); 5.9(d, 1H); 6.05(m, 1H); 7.1(s, 1H);7.25(s, 1H);7.25(m, 1H); 8.35(d, 1H); 8.4(m, 1H); 8.9(m, 1H).4. Reaction as in Footnote 3. Eluant proportion 20-25:80-75:1. N.m.r. insolvent B:1.15(t, 3H); 1.55(s, 6H); 3.4(q, 2H); 3.4-3.8(m, 4H); 4.4-5.2(m, 4H);5.15(d, 1H);5.95(d, 1H); 6.95(d, 1H); 7.05(s, 1H); 8.35(d, 1H).5. Reaction as in Footnote 3. Eluant proportion 20:80:1. N.m.r. insolvent B: 1.55(s,6H); 2.7-3.0(m, 2H); 3.3-4.2(m, 6H); 4.4-4.8(m, 3H); 5.0-5.3(m, 2H);5.85(d, 1H);7.0(s, 1H); 7.15(s, 1H); 8.45(d, 1H).6. Reaction as in Footnote 1, for 2 hr. at room temperature. Eluantproportion as inFootnote 4. N.m.r. in solvent B: 1.55(s, 6H); 3.55(s, 2H); 4.55(s, 2H);4.7(s, 3H);5.2(d, 1H); 5.85(d, 1H); 7.05(s, 1H); 7.15(d, 1H); 7.35(d, 1H); 8.05(t,1H); 8.85(d,1H); 9.35(d, 1H).7. The starting material quarternary heterocyclic salt was obtained from5-amino-cinnoline and methyl iodide. N.m.r. in solvent B: 4.65(s, 3H); 7.2(d,1H); 7.25(d,1H); 7.95(t, 1H); 8.9(d, 1H); 9.25(d, 1H).8. Eluant was MeOH/aqueous ammonium carbonate buffer, 1.3:8.7 v/v. N.m.r.insolvent B; 1.55(s, 6H); 2.7-2.9(m, 2H); 3.3-3.7(m, 2H); 3.8-4.1(m, 2H);4.6-5.0(m,4H); 5.2(d, 1H); 5.2-5.4(m, 2H); 5.9(d, 1H); 7.0(s, 1H); 7.3(d, 1H);8.3(d, 1H).
EXAMPLE 255
The process described in Example 61, Footnote 13 was repeated, using 7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxycyclobutyloxyimino)acetamido]-3-ethyl-aminoethylceph-3-em-4-carboxylic acid as the cephem starting material, to give 3-[N-(1-[2-aminoethyl]-4-pyridinio)-N-ethylaminomethyl]-7-[2-((Z)-1-carboxycyclobutyloxyimino)acetamidolceph-3-em-4-carboxylic acid. The product was purified by MPLC on a HP20SS column eluting with H.sub.2 O/CH.sub.3 CN, 100-93:0-7. N.m.r. in solvent A: 1.1 (t, 3H); 1.85 (m, 2H); 2.35 (m, 4H); 3.05 (d, 1H); 3.3 (m, 2H); 3.45 (d, 1H); 3.55-3.8 (m, 2H); 4.35-4.5 (m, 3H); 4.8 (d, 1H); 5.05 (d, 1H); 5.7 (d, 1H); 6.75 (s, 1H); 7.1 (m, 1H); 7.65 (m, 1H); 8.2 (d, 2H).
EXAMPLE 256
The general process described in Example 6 was repeated, using the appropriate heterocyclic starting material and cephem derivative, to give the following compound in 31% yield: ##STR296##
HPLC eluant MeOH/water/HOAc, 30-35:69-64:1 v/v/v.
N.m.r. in solvent B: 1.3 (t, 3H); 1.7 (m, 4H); 2.15 (m, 4H); 2.95 (q, 2H); 3.55 (m, 2H); 4.2 (d, 1H); 4.6 (d, 1H); 5.1 (d, 1H); 5.85 (d, 1H); 7.05 (s, 1H); 7.45 (d, 1H); 8.1 (s, 1H); 8.55 (d, 1H).
The starting material was 6-chloro-3-ethylthiazolo[3,2-b]pyridazinium perchlorate which was prepared by the general procedure described in J. Org. Chem., 34, 996, (1964).
EXAMPLE 257
The process described in Example 247 was repeated, using the appropriate 3-acetoxymethyl cephem and 5-amino-2-methylisoquinolinium iodide as starting materials. The reaction was carried out in water at pH 7.5 (adjusted by addition of HOAC or NaHCO.sub.3 as necessary), for 2.5 hrs. at 80.degree.. HPLC eluant was MeOH/H.sub.2 O/HOAc, 30-35:69-64:1 v/v/v. N.m.r. in solvent B: 1.7 (m, 4H); 2.15 (m, 4H); 3.6 (m, 2H); 4.4 (m, 5H); 5.15 (d, 1H); 5.85 (d, 1H); 7.05 (s, 1H); 7.15 (d, 1H); 7.5 (d, 1H); 7.6 (dd, 1H); 7.75 (dd, 1H); 8.5 (d, 1H); 8.65 (d, 1H); 9.75 (s, 1H).
The cephalosporin starting material may be obtained as follows:
To a stirred mixture of DMF (0.85 ml) in anhydrous CH.sub.2 Cl.sub.2 (40 ml) at -10.degree. C. was added dropwise a solution of oxalyl chloride (0.87 ml) in anhydrous CH.sub.2 Cl.sub.2 (10 ml). Stirring was continued at -10.degree. C. for 30 minutes. To this stirred suspension was added in one portion 2-[(Z)-(tert-butoxycarbonyl)cyclopentyloxyimino]- 2-(2-tritylaminothiazol-4-yl)acetic acid (5.97 g) followed by N-methylmorpholine (1.32 ml). Stirring was continued for 30 minutes at -10.degree. C.
In another flask a suspension of 7-aminocephalosporanic acid (2.72 g) in anhydrous CH.sub.2 Cl.sub.2 (25 ml) was stirred for 1 hour with N,O-bis (trimethylsilyl) acetamide (4 ml) to give a clear solution. This was transferred by syringe to the above acid chloride solution which was stirred at -10.degree. C. during the addition. The reaction mixture was then allowed to warm to room temperature and stirred for a further 1.5 hours. The reaction mixture was poured into water and extracted with CH.sub.2 Cl.sub.2. The combined CH.sub.2 Cl.sub.2 extracts were washed with water, dried (MgSO.sub.4) and the solvent was evaporated under reduced pressure to yield a foam. The crude product was dissolved in EtOAc and applied to a column of Kieselgel 60. Elution with EtOAc gave, after precipitation from CH.sub.2 Cl.sub.2 with n-hexane, 3-acetoxymethyl-7-[2-(2-tritylaminothiazol-4-yl)-2-((Z)-1-tert-butoxycarbonyl)cyclopentyloxyimino)acetamido]ceph-3-em-4-carboxylic acid (6.6g) as a white solid. This compound, (2.61g) was then stirred for 1 hour at 0.degree. C. with a mixture of TFA (12 ml) and water (1.2 ml). The reaction mixture was evaporated to dryness, the residue taken up in water and extracted with EtOAc. The solvent was evaporated under reduced pressure to give a solid which was dissolved in a minimum MEOH and precipitated with ether to give 3-acetoxymethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxy-1-cyclopentyloxyimino)acetamido]ceph-3-em-4-carboxylic acid (1.1 g) as a white solid having the following n.m.r. in solvent B: 1.7 (m, 3H); 1.95 (s, 3H); 2.15 (m, 4H); 3.4 (d, 1H); 3.65 (d, 1H); 4.7 (d, 1H); 4.95 (d, 1H); 5.15 (d, 1H); 5.85 (d, 1H); 7.05 (d, 1H).
EXAMPLE 258
Method A of Example 108 was repeated, using the appropriate starting materials, and water as solvent to give the compound: ##STR297## in 17.5% yield. N.m.r. in solvent B: 2.4-2.7 (m, 4H); 3.4-3.85 (ABq, 2H); 4.3-4.7 (Abq, 2H); 5.25 (d, 1H); 5.55 (s, 2H); 5.9 (d, 1H); 6.95 (s, 1H); 7.15 (d, 1H); 7.35-7.5 (m, 1H); 7.6 (d, 2H); 8.05 (d, 2H); 8.3 (d, 1H); after MPLC on HP20SS (as the sodium salt), and precipitation from aqueous solution with acetic acid.
The starting quaternary pyridinium salt was the tetrafluoroborate described in Example 243, Footnote 16.
EXAMPLE 259
The process described in Example 221 was repeated, using the appropriate 3-ethylaminomethyl cephem and 1-methyl-4-methylthioquinazolinium iodide, to give ##STR298##
n.m.r. in solvent A: 1.4 (br s, 9H); 3.1-3.5 (dd, 2H); 3.98 (br s, 5H); 5.04 (d, 1H); 5.1-5.5 (m, 2H); 5.68 (d, 1H); 6.71 (s, 1H); 7.8 (t, 1H); 7.99 (d, 1H); 8.12 (t, 1H); 8.28 (d, 1H); 8.85 (s, 1H).
EXAMPLE 260
The process described in Example 236 was repeated, using the appropriate 3-aminomethyl cephem and 1-furfuryl-4-methylsulphinylpyridinium chloride, to give in 20% yield, ##STR299##
n.m.r. in solvent A: 1.42 (2xs 6H); 3.35 (q, 2H); 4.3 (q, 2H); 5.01 (d, 1H); 5.37 (s, 2H); 5.67 (d, 1H); 6.45 (q, 1H); 6.62 (d, 1H); 6.73 (s, 1H); 6.93 (m, 1H); 7.4 (m, 1H); 7.64 (m, 1H); 8.12 (d, 1H); 8.28 (d, 1H).
The starting pyridinium salt was obtained by the general process described in Example 237, Footnote 4. The 1-furfuryl-4-methylthiopyridinium chloride had n.m.r. in d.sub.6 -DMSO: 2.72 (s, 3H); 5.92 (s, 2H); 6.52 (q, 1H); 6.84 (d, 1H); 7.75 (m, 1H); 8.0 (d, 2H); 8.96 (d, 2H). This compound was oxidised to the required sulphoxide, as described in Example 237, Footnote 4, and used without further purification.
EXAMPLE 261
The process described in Examples 1 to 4 was repeated, using the appropriate 3-ethylaminomethyl cephem and 3-chloro-1-methylpyridazinium iodide. The reaction was carried out for 18 hrs. at room temperature before quenching with HOAc, and isolating the product by chromatography on Diaion CHP20P resin, to give, in 51% yield, the more polar isomer of ##STR300##
n.m.r. spectrum in solvent A: 0.93 (t, 3H); 1.1 (t, 3H); 1.78 (quintet, 2H); 3.2 (d, 1H); 3.43 (d, 1H); 3.64 (m, 2H); 4.29 (s, 3H); 4.43 (t, 1H); 4.55 (d, 1H); 4.69 (d, 1H); 5.04 (d, 1H); 5.73 (d, 1H); 6.76 (s, 1H); 8.05 (m, 2H); 8.93 (t, 1H). Retention time 5.3 minutes on a reversed phase Partisil PXS 10/25 ODS-2 column, using MeOH/water/TFA 40:60:0.2 v/v/v as eluant.
The cephalosporin starting material may be prepared as follows: To a stirred solution of 3-aminomethyl-7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxypropoxyimino)acetamido]ceph-3-em-4-carboxylic acid (more polar isomer, 2.6g) in 2.5% (w/v) pH 5.5 sodium acetate buffer (50 ml) was added sodium cyanoborohydride (0.34 g), followed by acetaldehyde (0.3 ml) in water (5 ml) over 40 minutes. The reaction mixture was then concentrated, diluted to 50 ml with water, and the pH adjusted to 3.5 with HOAc. Careful chromatography on Diaion HP20SS, eluting with H.sub.2 O/CH.sub.3 CN, (92.5:7.5) gave 7-[2-(2-aminothiazol-4-yl)-2-((Z)-1-carboxypropoxyimino)acetamido]-3-ethylaminomethylceph-3-em-4-carboxylic acid, having the following n.m.r. spectrum in solvent A: 0.96 (t, 3H); 1.18 (t, 3H); 1.81 (quintet, 2H); 2.94 (q, 2H); 3.38 (d, 1H); 3.42 (d, 1H); 3.61 (d, 1H); 3.78 (d, 1H), 4.46 (t, 1H); 5.03 (d, 1H); 5.76 (d, 1H); 6.88 (s, 1H). Retention time 4.2 minutes on a reversed phase Partisil PXS 10/25 ODS-2 column, using MeOH/H.sub.2 O/HOAc,,30:70:1, v/v/v, as eluant.
EXAMPLE 262
The general process described in Example 244 was repeated, using 1-(4-carbamoylbenzyl)-4-chloropyridinium bromide as the starting material, to give ##STR301##
n.m.r. in solvent A: 1.05-1.2 (t, 3H); 1.4 (2xs, 6H); 3.2-3.7 (m, 4H); 4.45-4.7 (q, 2H); 5.15 (d, 1H); 5.45 (s, 2H); 5.8 (d, 1H); 6.7 (s, 1H); 7.0-7.4 (m, 2H); 7.4-7.5 (d, 2H); 7.85-7.95 (d, 2H); 8.35-8.45 (d, 2H).
The starting pyridinium salt was obtained by quaternising 4-chloropyridine with 4-bromomethylbenzamide in DMF at room temperature for 4 hours.
EXAMPLES 263-269
The general procedure described in Examples 1-4 was repeated, except that the solvent was DMF and the base was Et.sub.3 N. The appropriate 3-ethylaminomethylcephalosporin was used, and products were purified by chromatography on Diaion CHP20P resin. Retention times are quoted for a reversed phase Partisil PXS 10/25 ODS-2 column.
The following compounds were made:
__________________________________________________________________________ ##STR302## YieldExample R.sub.1 R.sub.2 (%) Footnotes__________________________________________________________________________263 ##STR303## ##STR304## 19 1, 2, 3264 ##STR305## ##STR306## 5 1, 4, 5265 ##STR307## ##STR308## 12 6, 7, 8266 ##STR309## ##STR310## 60 7, 9, 10267 ##STR311## ##STR312## 50 7, 11, 12268 ##STR313## ##STR314## 50 13, 14, 15269 ##STR315## ##STR316## 56 16, 17, 18__________________________________________________________________________1. Starting material was 6-amino-1-methylpyridazinium tetrafluoroborate.2. Reaction mixture heated 24 hours at 45.degree. C. before quenchingwith acetic acid.3. N.m.r. in solvent A: 0.93(t, 3H), 1.04(t, 3H), 1.8(m, 2H), 3.27(d,1H), 3.44(d, 1H), 3.48(m,2H), 3.75(s, 3H), 4.32(d, 1H), 4.47(t, 1H), 4.57(d, 1H), 5.07(d, 1H),5.88(d, 1H), 6.88(s, 1H),7.36(d, 1H), 7.75(d, 1H). Retention time 4.6 minutes, eluting withMeOH/H.sub.2 O/HOAc, 40:60:1,v/v/v.4. Reaction mixture heated at 65.degree. C. for 3 hours before quenchingwith acetic acid. After apreliminary purification on Diaion CHP20P, the product was re-purified bypreparative HPLCon a reversed phase Partisil ODS2 column, eluting in MeOH/H.sub.2 O/HOAc,75:25:1 v/v/v.5. N.m.r. in solvent A: 1.05(t, 3H), 1.39(d, 3H), 3.24(d, 1H), 3.43(d,1H), 3.48(m, 2H), 3.74(s,3H), 4.31(d, 1H), 4.53(d, 1H), 4.6(q, 1H), 5.07(d, 1H), 5.75(d, 1H),6.87(s, 1H), 7.33(d, 1H),7.88(d, 1H). Retention time 4.5 minutes, eluting with MeOH/water/HOAc,65:35:1, v/v/v.6. Starting material 6-fluoro-thiazolo[3,2-b]pyridazinium perchlorate.7. Reaction carried out at ambient temperature for 18 hours beforequenching with HOAc.8. N.m.r. in solvent A: 0.92(t, 3H), 1.15(t, 3H), 1.79(m, 2H), 3.32(d,1H), 3.51(d, 1H), 3.68(m,2H), 4.45(t, 1H), 4.58(d, 1H), 4.79(d, 1H), 5.05(d, 1H), 5.77(d, 1H),6.76(s, 1H), 7.89(d, 1H),8.46(d, 1H), 8.76(d, 1H), 8.78(d, 1H). Retention time 4.7 minutes,eluting with MeOH/H.sub.2 O/HOAc, 40:60:1, v/v/v.9. Starting material was 1-methyl-4-methylthiothieno[2,3-d]pyrimidiniumtetrafluoroborate.10. N.m.r. in solvent A: 0.9(t, 3H), 1.3(m, 3H), 1.77(m, 2H), 3.24(d,1H), 3.46(d, 1H), 3.98(m,2H), 4.0(s, 3H), 4.44(t, 1H), 4.84(d, 1H), 5.06(d, 1H), 5.25(d, 1H),5.77(d, 1H), 6.76(s, 1H),7.76(d, 1H), 7.87(d, 1H), 8.77(s, 1H). Retention time 4.1 minutes,eluting with MeOH/H.sub.2 O/TFA, 45:65:0.1, v/v/v.11. Starting material was 6,7-dihydro-2-methylthio-thiazolo [3,2-a]pyrimidinium tetrafluoro-borate.12. N.m.r. in solvent A: 1.11(t, 3H), 1.39(d, 3H), 3.16(d, 1H), 3.43(d,1H), 3.66(m, 4H), 4.6(m,4H), 4.96(d, 1H), 5.06(d, 1H), 5.78(d, 1H), 6.77(s, 1H), 6.83(d, 1H),8.23(d, 1H). Retention time5.0 minutes, eluting with MeOH/H.sub.2 O/TFA, 40:60:0.2, v/v/v.13. Prepared by the standard aqueous/DMF conditions of Examples 1-4,except that thereaction was carried out for 65 hours at ambient temperature beforequenching with HOAc.14. Starting material was 1-allyl-4-chloropyridinium tosylate.15. N.m.r. in solvent A: 1.12(t, 3H), 1.39(d, 3H), 3.15(d, 1H), 3.45(d,1H), 3.63(m, 2H), 4.5(d,1H), 4.58(q, 1H), 4.71(d, 1H), 4.78(d, 2H), 5.06(d, 1H), 5.25(d, 1H),5.33(d, 1H), 5.77(d, 1H),6.03(m, 1H), 6.75(s, 1H), 7.1(br, 1H), 7.4(br, 1H), 8.19(d, 2H).Retention time 3.9 minutes,eluting with MeOH/H.sub.2 O/TFA, 40:60:0.1, v/v/v.16. Prepared using the procedure of Footnote 13 above, except that thecrude product waschromatographed with out acidification on CHP20P, to give the product asa mono-sodiumsalt, after precipitation from water by acetone.17. Starting material was 4-chloro-1-(4-nitrobenzyl)-pyridiniumtosylate.18. N.m.r. in solvent A: 1.11(t, 3H), 1.37(d, 3H), 3.1(d, 1H), 3.51(d,1H), 3.76(m, 2H), 4.48(d,1H), 4.57(q, 1H), 4.77(d, 1H), 5.0(d, 1H), 5.52(s, 2H), 5.7(d, 1H),6.75(s, 1H), 7.1(br, 1H),7.55(br, 1H), 7.63(d, 2H), 8.25(d, 2H), 8.36(d, 2H). Retention time 8.5minutes in MeOH/H.sub.2 O/TFA, 40:60:0.2, v/v/v. ##STR317##

Number	Name	Date	Kind
5049558	Bradbury et al.	Sep 1991
5149803	Davies et al.	Sep 1992

Cephalosporin intermediates

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Chemical Abstracts vol. 113(1): 6033m (1989).
Chemical Abstracts vol. 106(3): 18243y (1987).