NO171271B - QUATERNARY AMINTIOL COMPOUNDS - Google Patents
QUATERNARY AMINTIOL COMPOUNDS Download PDFInfo
- Publication number
- NO171271B NO171271B NO902554A NO902554A NO171271B NO 171271 B NO171271 B NO 171271B NO 902554 A NO902554 A NO 902554A NO 902554 A NO902554 A NO 902554A NO 171271 B NO171271 B NO 171271B
- Authority
- NO
- Norway
- Prior art keywords
- mmol
- carboxylate
- pyrldinium
- hept
- oxo
- Prior art date
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- 150000001875 compounds Chemical class 0.000 title description 43
- 229910052757 nitrogen Inorganic materials 0.000 claims description 28
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 18
- 125000000217 alkyl group Chemical group 0.000 claims description 17
- -1 amine thiol compounds Chemical class 0.000 claims description 13
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 11
- SJRJJKPEHAURKC-UHFFFAOYSA-N N-Methylmorpholine Chemical compound CN1CCOCC1 SJRJJKPEHAURKC-UHFFFAOYSA-N 0.000 claims description 9
- 125000004433 nitrogen atom Chemical group N* 0.000 claims description 8
- RXYPXQSKLGGKOL-UHFFFAOYSA-N 1,4-dimethylpiperazine Chemical group CN1CCN(C)CC1 RXYPXQSKLGGKOL-UHFFFAOYSA-N 0.000 claims description 4
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 4
- KDTVWEHAAISPNW-UHFFFAOYSA-N 4-methylthiomorpholine Chemical compound CN1CCSCC1 KDTVWEHAAISPNW-UHFFFAOYSA-N 0.000 claims description 3
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 claims description 2
- 239000005977 Ethylene Substances 0.000 claims description 2
- MTNDZQHUAFNZQY-UHFFFAOYSA-N imidazoline Chemical compound C1CN=CN1 MTNDZQHUAFNZQY-UHFFFAOYSA-N 0.000 claims description 2
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 189
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 143
- 229910001868 water Inorganic materials 0.000 description 108
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 95
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 80
- 239000000243 solution Substances 0.000 description 74
- 239000000203 mixture Substances 0.000 description 70
- 238000005160 1H NMR spectroscopy Methods 0.000 description 41
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 40
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 38
- 239000011541 reaction mixture Substances 0.000 description 38
- 238000004108 freeze drying Methods 0.000 description 37
- 239000007787 solid Substances 0.000 description 37
- 239000012071 phase Substances 0.000 description 32
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 31
- 125000003118 aryl group Chemical group 0.000 description 30
- 238000005481 NMR spectroscopy Methods 0.000 description 27
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 25
- 125000004005 formimidoyl group Chemical group [H]\N=C(/[H])* 0.000 description 25
- 125000004432 carbon atom Chemical group C* 0.000 description 24
- 239000008363 phosphate buffer Substances 0.000 description 24
- DBTDEFJAFBUGPP-UHFFFAOYSA-N Methanethial Chemical compound S=C DBTDEFJAFBUGPP-UHFFFAOYSA-N 0.000 description 23
- 239000000843 powder Substances 0.000 description 23
- 239000007864 aqueous solution Substances 0.000 description 22
- 239000003921 oil Substances 0.000 description 22
- 239000003960 organic solvent Substances 0.000 description 22
- 238000002360 preparation method Methods 0.000 description 22
- 239000000047 product Substances 0.000 description 22
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 21
- 238000010828 elution Methods 0.000 description 21
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium Chemical compound [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 20
- BHIIGRBMZRSDRI-UHFFFAOYSA-N [chloro(phenoxy)phosphoryl]oxybenzene Chemical compound C=1C=CC=CC=1OP(=O)(Cl)OC1=CC=CC=C1 BHIIGRBMZRSDRI-UHFFFAOYSA-N 0.000 description 20
- JUJWROOIHBZHMG-UHFFFAOYSA-O pyridinium Chemical compound C1=CC=[NH+]C=C1 JUJWROOIHBZHMG-UHFFFAOYSA-O 0.000 description 20
- 150000003955 ε-lactams Chemical class 0.000 description 19
- 125000004705 ethylthio group Chemical group C(C)S* 0.000 description 18
- 125000000623 heterocyclic group Chemical group 0.000 description 18
- VOVUARRWDCVURC-UHFFFAOYSA-N thiirane Chemical compound C1CS1 VOVUARRWDCVURC-UHFFFAOYSA-N 0.000 description 18
- 239000010410 layer Substances 0.000 description 16
- 238000004458 analytical method Methods 0.000 description 15
- 239000008346 aqueous phase Substances 0.000 description 15
- 239000000872 buffer Substances 0.000 description 15
- 229910052739 hydrogen Inorganic materials 0.000 description 15
- 239000001257 hydrogen Substances 0.000 description 15
- 238000000034 method Methods 0.000 description 14
- 150000007942 carboxylates Chemical class 0.000 description 13
- 150000002148 esters Chemical class 0.000 description 13
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 13
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 12
- 239000000460 chlorine Chemical group 0.000 description 12
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 12
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 11
- FYZUENZXIZCLAZ-UHFFFAOYSA-N 2-methylhept-2-enoic acid Chemical compound CCCCC=C(C)C(O)=O FYZUENZXIZCLAZ-UHFFFAOYSA-N 0.000 description 10
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N acetone Substances CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 10
- YZBQHRLRFGPBSL-RXMQYKEDSA-N carbapenem Chemical class C1C=CN2C(=O)C[C@H]21 YZBQHRLRFGPBSL-RXMQYKEDSA-N 0.000 description 10
- 125000000753 cycloalkyl group Chemical group 0.000 description 10
- 239000012299 nitrogen atmosphere Substances 0.000 description 10
- 239000012044 organic layer Substances 0.000 description 10
- 239000006188 syrup Substances 0.000 description 10
- 235000020357 syrup Nutrition 0.000 description 10
- GAWIXWVDTYZWAW-UHFFFAOYSA-N C[CH]O Chemical compound C[CH]O GAWIXWVDTYZWAW-UHFFFAOYSA-N 0.000 description 9
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 9
- 238000001816 cooling Methods 0.000 description 9
- 229940098779 methanesulfonic acid Drugs 0.000 description 9
- 239000000725 suspension Substances 0.000 description 9
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 8
- 229910052760 oxygen Inorganic materials 0.000 description 8
- 125000001424 substituent group Chemical group 0.000 description 8
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 8
- MOOOYQDIYFVYGD-UHFFFAOYSA-J tripotassium;sodium;hydroxide;phosphate Chemical compound [OH-].[Na+].[K+].[K+].[K+].[O-]P([O-])([O-])=O MOOOYQDIYFVYGD-UHFFFAOYSA-J 0.000 description 8
- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 description 7
- ASMQGLCHMVWBQR-UHFFFAOYSA-M diphenyl phosphate Chemical compound C=1C=CC=CC=1OP(=O)([O-])OC1=CC=CC=C1 ASMQGLCHMVWBQR-UHFFFAOYSA-M 0.000 description 7
- 239000003480 eluent Substances 0.000 description 7
- 239000000706 filtrate Substances 0.000 description 7
- FKNQCJSGGFJEIZ-UHFFFAOYSA-N 4-methylpyridine Chemical compound CC1=CC=NC=C1 FKNQCJSGGFJEIZ-UHFFFAOYSA-N 0.000 description 6
- 125000003710 aryl alkyl group Chemical group 0.000 description 6
- 125000004429 atom Chemical group 0.000 description 6
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 125000006239 protecting group Chemical group 0.000 description 6
- 238000004007 reversed phase HPLC Methods 0.000 description 6
- 239000000741 silica gel Substances 0.000 description 6
- 229910002027 silica gel Inorganic materials 0.000 description 6
- 238000003756 stirring Methods 0.000 description 6
- 239000003643 water by type Substances 0.000 description 6
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 5
- 229910019142 PO4 Inorganic materials 0.000 description 5
- CSCPPACGZOOCGX-WFGJKAKNSA-N acetone d6 Chemical compound [2H]C([2H])([2H])C(=O)C([2H])([2H])[2H] CSCPPACGZOOCGX-WFGJKAKNSA-N 0.000 description 5
- 125000001931 aliphatic group Chemical group 0.000 description 5
- 125000003342 alkenyl group Chemical group 0.000 description 5
- 125000000304 alkynyl group Chemical group 0.000 description 5
- 150000001412 amines Chemical class 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
- 125000001743 benzylic group Chemical group 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 125000001316 cycloalkyl alkyl group Chemical group 0.000 description 5
- 238000001704 evaporation Methods 0.000 description 5
- 230000008020 evaporation Effects 0.000 description 5
- 125000004475 heteroaralkyl group Chemical group 0.000 description 5
- 125000001072 heteroaryl group Chemical group 0.000 description 5
- 125000005842 heteroatom Chemical group 0.000 description 5
- 125000004415 heterocyclylalkyl group Chemical group 0.000 description 5
- 125000000250 methylamino group Chemical group [H]N(*)C([H])([H])[H] 0.000 description 5
- 239000001301 oxygen Substances 0.000 description 5
- 239000010452 phosphate Substances 0.000 description 5
- 125000004434 sulfur atom Chemical group 0.000 description 5
- BSKHPKMHTQYZBB-UHFFFAOYSA-N 2-methylpyridine Chemical compound CC1=CC=CC=N1 BSKHPKMHTQYZBB-UHFFFAOYSA-N 0.000 description 4
- CUYKNJBYIJFRCU-UHFFFAOYSA-N 3-aminopyridine Chemical compound NC1=CC=CN=C1 CUYKNJBYIJFRCU-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 150000001450 anions Chemical class 0.000 description 4
- 125000000113 cyclohexyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 description 4
- QJFUMFCCMJJLIE-UHFFFAOYSA-N ethanethial Chemical compound CC=S QJFUMFCCMJJLIE-UHFFFAOYSA-N 0.000 description 4
- 239000012065 filter cake Substances 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 4
- CBOIHMRHGLHBPB-UHFFFAOYSA-N hydroxymethyl Chemical compound O[CH2] CBOIHMRHGLHBPB-UHFFFAOYSA-N 0.000 description 4
- 230000014759 maintenance of location Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- PTMBWNZJOQBTBK-UHFFFAOYSA-N pyridin-4-ylmethanol Chemical compound OCC1=CC=NC=C1 PTMBWNZJOQBTBK-UHFFFAOYSA-N 0.000 description 4
- ITMCEJHCFYSIIV-UHFFFAOYSA-N triflic acid Chemical compound OS(=O)(=O)C(F)(F)F ITMCEJHCFYSIIV-UHFFFAOYSA-N 0.000 description 4
- 239000003039 volatile agent Substances 0.000 description 4
- OISVCGZHLKNMSJ-UHFFFAOYSA-N 2,6-dimethylpyridine Chemical compound CC1=CC=CC(C)=N1 OISVCGZHLKNMSJ-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical group [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 3
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 125000001118 alkylidene group Chemical group 0.000 description 3
- 125000000129 anionic group Chemical group 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 229910052801 chlorine Inorganic materials 0.000 description 3
- UAOMVDZJSHZZME-UHFFFAOYSA-N diisopropylamine Chemical compound CC(C)NC(C)C UAOMVDZJSHZZME-UHFFFAOYSA-N 0.000 description 3
- 239000012156 elution solvent Substances 0.000 description 3
- 150000002431 hydrogen Chemical class 0.000 description 3
- IXCSERBJSXMMFS-UHFFFAOYSA-N hydrogen chloride Substances Cl.Cl IXCSERBJSXMMFS-UHFFFAOYSA-N 0.000 description 3
- 229910000041 hydrogen chloride Inorganic materials 0.000 description 3
- 239000012074 organic phase Substances 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 3
- 125000003367 polycyclic group Chemical group 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 229920006395 saturated elastomer Polymers 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- AULOGPOZBOCNSX-UHFFFAOYSA-N 3-methylsulfanylpyridine Chemical compound CSC1=CC=CN=C1 AULOGPOZBOCNSX-UHFFFAOYSA-N 0.000 description 2
- MVQVNTPHUGQQHK-UHFFFAOYSA-N 3-pyridinemethanol Chemical compound OCC1=CC=CN=C1 MVQVNTPHUGQQHK-UHFFFAOYSA-N 0.000 description 2
- XCHSSZMKKXPBAI-UHFFFAOYSA-N 4-methylsulfanylpyridine Chemical compound CSC1=CC=NC=C1 XCHSSZMKKXPBAI-UHFFFAOYSA-N 0.000 description 2
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 2
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical group [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 2
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 2
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 2
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical group FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 125000005119 alkyl cycloalkyl group Chemical group 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Chemical group BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 2
- 229910052794 bromium Chemical group 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000012230 colorless oil Substances 0.000 description 2
- 238000004440 column chromatography Methods 0.000 description 2
- 238000002425 crystallisation Methods 0.000 description 2
- 230000008025 crystallization Effects 0.000 description 2
- 125000004956 cyclohexylene group Chemical group 0.000 description 2
- 125000004979 cyclopentylene group Chemical group 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- SHZIWNPUGXLXDT-UHFFFAOYSA-N ethyl hexanoate Chemical compound CCCCCC(=O)OCC SHZIWNPUGXLXDT-UHFFFAOYSA-N 0.000 description 2
- 239000011737 fluorine Chemical group 0.000 description 2
- 229910052731 fluorine Inorganic materials 0.000 description 2
- 210000003918 fraction a Anatomy 0.000 description 2
- 229910052736 halogen Inorganic materials 0.000 description 2
- 150000002367 halogens Chemical group 0.000 description 2
- 239000008240 homogeneous mixture Substances 0.000 description 2
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 2
- 239000002198 insoluble material Substances 0.000 description 2
- 239000013067 intermediate product Substances 0.000 description 2
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 description 2
- 150000003951 lactams Chemical class 0.000 description 2
- 238000002844 melting Methods 0.000 description 2
- 230000008018 melting Effects 0.000 description 2
- IHJIGXQQHGGSEE-UHFFFAOYSA-N methanesulfonate;2-pyridin-1-ium-1-ylethanethiol Chemical compound CS([O-])(=O)=O.SCC[N+]1=CC=CC=C1 IHJIGXQQHGGSEE-UHFFFAOYSA-N 0.000 description 2
- BLTAPEIEHGWKKN-UHFFFAOYSA-N methanesulfonate;pyridin-1-ium Chemical compound CS(O)(=O)=O.C1=CC=NC=C1 BLTAPEIEHGWKKN-UHFFFAOYSA-N 0.000 description 2
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 2
- OVXDLDSWRLLUJT-UHFFFAOYSA-N morpholine;trifluoromethanesulfonic acid Chemical compound C1COCC[NH2+]1.[O-]S(=O)(=O)C(F)(F)F OVXDLDSWRLLUJT-UHFFFAOYSA-N 0.000 description 2
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 2
- ZUFQCVZBBNZMKD-UHFFFAOYSA-M potassium 2-ethylhexanoate Chemical compound [K+].CCCCC(CC)C([O-])=O ZUFQCVZBBNZMKD-UHFFFAOYSA-M 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- AOJFQRQNPXYVLM-UHFFFAOYSA-N pyridin-1-ium;chloride Chemical compound [Cl-].C1=CC=[NH+]C=C1 AOJFQRQNPXYVLM-UHFFFAOYSA-N 0.000 description 2
- 125000001453 quaternary ammonium group Chemical group 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 150000003573 thiols Chemical class 0.000 description 2
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 2
- RIOQSEWOXXDEQQ-UHFFFAOYSA-N triphenylphosphine Chemical compound C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RIOQSEWOXXDEQQ-UHFFFAOYSA-N 0.000 description 2
- YBIDYTOJOXKBLO-USLOAXSXSA-N (4-nitrophenyl)methyl (5r,6s)-6-[(1r)-1-hydroxyethyl]-3,7-dioxo-1-azabicyclo[3.2.0]heptane-2-carboxylate Chemical compound C([C@@H]1[C@H](C(N11)=O)[C@H](O)C)C(=O)C1C(=O)OCC1=CC=C([N+]([O-])=O)C=C1 YBIDYTOJOXKBLO-USLOAXSXSA-N 0.000 description 1
- ICHWPLYIJVROHH-QDBPVWLISA-N (5R,6S)-6-[(1R)-1-hydroxyethyl]-3-[2-(3-methylsulfanylpyridin-1-ium-1-yl)ethylsulfanyl]-4-[(4-nitrophenyl)methyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylic acid chloride Chemical compound [Cl-].[N+](=O)([O-])C1=CC=C(CC2C(=C(N3C([C@@H]([C@H]23)[C@@H](C)O)=O)C(=O)O)SCC[N+]2=CC(=CC=C2)SC)C=C1 ICHWPLYIJVROHH-QDBPVWLISA-N 0.000 description 1
- LKIVEIAXFZBGMO-UHFFFAOYSA-N 1-methyl-2-methylsulfanylimidazole Chemical compound CSC1=NC=CN1C LKIVEIAXFZBGMO-UHFFFAOYSA-N 0.000 description 1
- FHTDDANQIMVWKZ-UHFFFAOYSA-N 1h-pyridine-4-thione Chemical compound SC1=CC=NC=C1 FHTDDANQIMVWKZ-UHFFFAOYSA-N 0.000 description 1
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 description 1
- JXFBYXLGJXDTMJ-UHFFFAOYSA-N 2-(3,5-dimethylpyridin-1-ium-1-yl)ethanethiol;methanesulfonate Chemical compound CS([O-])(=O)=O.CC1=CC(C)=C[N+](CCS)=C1 JXFBYXLGJXDTMJ-UHFFFAOYSA-N 0.000 description 1
- RUPYJMSRDVEDTL-UHFFFAOYSA-N 2-(3-aminopyridin-1-ium-1-yl)ethanethiol;chloride Chemical compound [Cl-].NC1=CC=C[N+](CCS)=C1 RUPYJMSRDVEDTL-UHFFFAOYSA-N 0.000 description 1
- IOZQQSRUUCKOPB-UHFFFAOYSA-N 2-(3-methyl-2-methylsulfanylimidazol-3-ium-1-yl)ethanethiol;trifluoromethanesulfonate Chemical compound [O-]S(=O)(=O)C(F)(F)F.CSC=1N(C)C=C[N+]=1CCS IOZQQSRUUCKOPB-UHFFFAOYSA-N 0.000 description 1
- VSGDHUSOTRUPBU-UHFFFAOYSA-N 2-(3-methylsulfanylpyridin-1-ium-1-yl)ethanethiol;chloride Chemical compound [Cl-].CSC1=CC=C[N+](CCS)=C1 VSGDHUSOTRUPBU-UHFFFAOYSA-N 0.000 description 1
- RIBHPTFRWWJERH-UHFFFAOYSA-N 2-pyridin-1-ium-1-ylethanethiol;chloride Chemical compound [Cl-].SCC[N+]1=CC=CC=C1 RIBHPTFRWWJERH-UHFFFAOYSA-N 0.000 description 1
- NHQDETIJWKXCTC-UHFFFAOYSA-N 3-chloroperbenzoic acid Chemical compound OOC(=O)C1=CC=CC(Cl)=C1 NHQDETIJWKXCTC-UHFFFAOYSA-N 0.000 description 1
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 1
- RQIIHWBNPSTACF-PLHIQKDASA-N C[C@@H](O)[C@@H]1[C@H]2CC(=O)C(Cc3ccc(cc3)[N+]([O-])=O)(N2C1=O)C(O)=O Chemical compound C[C@@H](O)[C@@H]1[C@H]2CC(=O)C(Cc3ccc(cc3)[N+]([O-])=O)(N2C1=O)C(O)=O RQIIHWBNPSTACF-PLHIQKDASA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- LVIPRKRYIVPTPB-UHFFFAOYSA-N Cl.CCCCC=C(C)C(O)=O Chemical compound Cl.CCCCC=C(C)C(O)=O LVIPRKRYIVPTPB-UHFFFAOYSA-N 0.000 description 1
- 235000004694 Eucalyptus leucoxylon Nutrition 0.000 description 1
- 244000166102 Eucalyptus leucoxylon Species 0.000 description 1
- 239000012359 Methanesulfonyl chloride Substances 0.000 description 1
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 description 1
- DZIHOXKQCPWUPF-UHFFFAOYSA-N P(=O)(OC1=CC=CC=C1)(OC1=CC=CC=C1)O.C(C=CCCCC)C(=O)O Chemical compound P(=O)(OC1=CC=CC=C1)(OC1=CC=CC=C1)O.C(C=CCCCC)C(=O)O DZIHOXKQCPWUPF-UHFFFAOYSA-N 0.000 description 1
- FOIXSVOLVBLSDH-UHFFFAOYSA-N Silver ion Chemical compound [Ag+] FOIXSVOLVBLSDH-UHFFFAOYSA-N 0.000 description 1
- KPDYUHWAVMBMCY-UHFFFAOYSA-N [1-(2-sulfanylethyl)pyridin-1-ium-3-yl]methanol;trifluoromethanesulfonate Chemical compound [O-]S(=O)(=O)C(F)(F)F.OCC1=CC=C[N+](CCS)=C1 KPDYUHWAVMBMCY-UHFFFAOYSA-N 0.000 description 1
- XZNLLKQCCSKOAN-UHFFFAOYSA-N [1-(2-sulfanylethyl)pyridin-1-ium-4-yl]methanol;trifluoromethanesulfonate Chemical compound [O-]S(=O)(=O)C(F)(F)F.OCC1=CC=[N+](CCS)C=C1 XZNLLKQCCSKOAN-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 125000002947 alkylene group Chemical group 0.000 description 1
- 125000005365 aminothiol group Chemical group 0.000 description 1
- 239000006286 aqueous extract Substances 0.000 description 1
- 125000006615 aromatic heterocyclic group Chemical group 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000012267 brine Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 229940043279 diisopropylamine Drugs 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- ACWVAOJPKNFQCU-UHFFFAOYSA-N diphenyl hydrogen phosphate 2-methylhept-2-enoic acid Chemical compound CCCCC=C(C)C(O)=O.C=1C=CC=CC=1OP(=O)(O)OC1=CC=CC=C1 ACWVAOJPKNFQCU-UHFFFAOYSA-N 0.000 description 1
- YOSCKURVDXUBRV-OFPMPFOCSA-M diphenyl phosphate;(4-nitrophenyl)methyl (5r,6s)-3-[2-(3-aminopyridin-1-ium-1-yl)ethylsulfanyl]-6-[(1r)-1-hydroxyethyl]-7-oxo-1-azabicyclo[3.2.0]hept-2-ene-2-carboxylate Chemical compound C=1C=CC=CC=1OP(=O)([O-])OC1=CC=CC=C1.C([C@@H]1[C@H](C(N1C=1C(=O)OCC=2C=CC(=CC=2)[N+]([O-])=O)=O)[C@H](O)C)C=1SCC[N+]1=CC=CC(N)=C1 YOSCKURVDXUBRV-OFPMPFOCSA-M 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 235000019439 ethyl acetate Nutrition 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- MDQRDWAGHRLBPA-UHFFFAOYSA-N fluoroamine Chemical group FN MDQRDWAGHRLBPA-UHFFFAOYSA-N 0.000 description 1
- 210000002196 fr. b Anatomy 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- BICAGYDGRXJYGD-UHFFFAOYSA-N hydrobromide;hydrochloride Chemical compound Cl.Br BICAGYDGRXJYGD-UHFFFAOYSA-N 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- ILVUABTVETXVMV-UHFFFAOYSA-N hydron;bromide;iodide Chemical compound Br.I ILVUABTVETXVMV-UHFFFAOYSA-N 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-M iodide Chemical compound [I-] XMBWDFGMSWQBCA-UHFFFAOYSA-M 0.000 description 1
- 229940006461 iodide ion Drugs 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- LGIQVXJKJREQSH-UHFFFAOYSA-N methanesulfonate;2-(2-methylpyridin-1-ium-1-yl)ethanethiol Chemical compound CS([O-])(=O)=O.CC1=CC=CC=[N+]1CCS LGIQVXJKJREQSH-UHFFFAOYSA-N 0.000 description 1
- ZQTFSGQLSHPNHC-UHFFFAOYSA-N methanesulfonate;2-(3-methoxypyridin-1-ium-1-yl)ethanethiol Chemical compound CS([O-])(=O)=O.COC1=CC=C[N+](CCS)=C1 ZQTFSGQLSHPNHC-UHFFFAOYSA-N 0.000 description 1
- APDAYUCAGWWUSR-UHFFFAOYSA-N methanesulfonate;2-(4-methylpyridin-1-ium-1-yl)ethanethiol Chemical compound CS([O-])(=O)=O.CC1=CC=[N+](CCS)C=C1 APDAYUCAGWWUSR-UHFFFAOYSA-N 0.000 description 1
- CJXRIDPGXABNQN-UHFFFAOYSA-N methanesulfonate;2-(4-methylsulfanylpyridin-1-ium-1-yl)ethanethiol Chemical compound CS([O-])(=O)=O.CSC1=CC=[N+](CCS)C=C1 CJXRIDPGXABNQN-UHFFFAOYSA-N 0.000 description 1
- BAXZXTYYXNQCAZ-UHFFFAOYSA-N methanesulfonate;2-(4-methylthiomorpholin-4-ium-4-yl)ethanethiol Chemical compound CS([O-])(=O)=O.SCC[N+]1(C)CCSCC1 BAXZXTYYXNQCAZ-UHFFFAOYSA-N 0.000 description 1
- GXWXBVSXQWXEHX-UHFFFAOYSA-N methanesulfonic acid;2-methylpyridine Chemical compound CS([O-])(=O)=O.CC1=CC=CC=[NH+]1 GXWXBVSXQWXEHX-UHFFFAOYSA-N 0.000 description 1
- QISWLEJRWWFHLF-UHFFFAOYSA-N methanesulfonic acid;4-methylpyridine Chemical compound CS([O-])(=O)=O.CC1=CC=[NH+]C=C1 QISWLEJRWWFHLF-UHFFFAOYSA-N 0.000 description 1
- QARBMVPHQWIHKH-UHFFFAOYSA-N methanesulfonyl chloride Chemical compound CS(Cl)(=O)=O QARBMVPHQWIHKH-UHFFFAOYSA-N 0.000 description 1
- 125000002816 methylsulfanyl group Chemical group [H]C([H])([H])S[*] 0.000 description 1
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 1
- IWPOSDLLFZKGOW-UHFFFAOYSA-N oct-3-enoic acid Chemical compound CCCCC=CCC(O)=O IWPOSDLLFZKGOW-UHFFFAOYSA-N 0.000 description 1
- NFHFRUOZVGFOOS-UHFFFAOYSA-N palladium;triphenylphosphane Chemical compound [Pd].C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 NFHFRUOZVGFOOS-UHFFFAOYSA-N 0.000 description 1
- 125000000843 phenylene group Chemical group C1(=C(C=CC=C1)*)* 0.000 description 1
- UXWKNNJFYZFNDI-UHFFFAOYSA-N piperazine;dihydrobromide Chemical compound Br.Br.C1CNCCN1 UXWKNNJFYZFNDI-UHFFFAOYSA-N 0.000 description 1
- AJZFEJQZIOIQSR-UHFFFAOYSA-M potassium;diphenyl phosphate Chemical compound [K+].C=1C=CC=CC=1OP(=O)([O-])OC1=CC=CC=C1 AJZFEJQZIOIQSR-UHFFFAOYSA-M 0.000 description 1
- LJSOLTRJEQZSHV-UHFFFAOYSA-L potassium;sodium;hydron;hydroxide;phosphate Chemical compound [OH-].[Na+].[K+].OP(O)([O-])=O LJSOLTRJEQZSHV-UHFFFAOYSA-L 0.000 description 1
- 238000002953 preparative HPLC Methods 0.000 description 1
- 238000005956 quaternization reaction Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- LROACRCVHZCGLL-UHFFFAOYSA-M s-[2-(1,4-dimethylpiperazin-1-ium-1-yl)ethyl] ethanethioate;bromide Chemical compound [Br-].CN1CC[N+](C)(CCSC(C)=O)CC1 LROACRCVHZCGLL-UHFFFAOYSA-M 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- SNOOUWRIMMFWNE-UHFFFAOYSA-M sodium;6-[(3,4,5-trimethoxybenzoyl)amino]hexanoate Chemical compound [Na+].COC1=CC(C(=O)NCCCCCC([O-])=O)=CC(OC)=C1OC SNOOUWRIMMFWNE-UHFFFAOYSA-M 0.000 description 1
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 125000000446 sulfanediyl group Chemical group *S* 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- ITMCEJHCFYSIIV-UHFFFAOYSA-M triflate Chemical compound [O-]S(=O)(=O)C(F)(F)F ITMCEJHCFYSIIV-UHFFFAOYSA-M 0.000 description 1
- 150000003952 β-lactams Chemical class 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Description
Foreliggende oppfinnelse angår kvaternære amintiolforbindelser. The present invention relates to quaternary amine thiol compounds.
Nærmere bestemt angår oppfinnelsen kvaternære amintiolforbindelser som karakteriseres ved at de har formelen More specifically, the invention relates to quaternary amine thiol compounds which are characterized by having the formula
der there
A er etylen som eventuelt er substituert med C1_4~alkyl, A is ethylene which is optionally substituted with C1_4~alkyl,
eller cykloheks-1,3-ylen og or cyclohex-1,3-ylene and
R<14> er et kvaternisert pyridin-, imidazolin-, N-metylmorfolin, N-metyltiomorfolin, N-metyltiomorfolin-s-oksyd-eller 1,4-dimetylpiperazingruppe som eventuelt er substituert med en eller flere grupper valgt blant C^g-alkyl, hydroksy-C^_g-alkyltio, C^_g-alkyloksy og amino og er bundet til A via det kvaternære nitrogenatom. R<14> is a quaternized pyridine, imidazoline, N-methylmorpholine, N-methylthiomorpholine, N-methylthiomorpholine-s-oxide or 1,4-dimethylpiperazine group which is optionally substituted with one or more groups selected from C^g- alkyl, hydroxy-C1-6-alkylthio, C1-6-alkyloxy and amino and is attached to A via the quaternary nitrogen atom.
Foreliggende oppfinnelse er avdelt fra NO-PS 166038 som beskriver en fremgangsmåte for fremstilling av karbapenem-derivater med formelen The present invention is separated from NO-PS 166038 which describes a method for the production of carbapenem derivatives with the formula
der there
R 8 er hydrogen, R 8 is hydrogen,
R<1> er l-(R)-hydroksyetyl, R<1> is 1-(R)-hydroxyethyl,
A og R<*4> har den ovenfor angitte betydning, og A and R<*4> have the meaning indicated above, and
R<2> er hydrogen, en anionisk blanding eller en konvensjonell lett avspaltbar karboksylbeskyttende gruppe under den forutsetning at det, når R<2> er hydrogen eller en beskyttende gruppe, også foreligger et motanion, eller R<2> is hydrogen, an anionic mixture or a conventional easily cleavable carboxyl protecting group provided that, when R<2> is hydrogen or a protecting group, a counter anion is also present, or
et farmasøytisk akseptabelt salt derav. a pharmaceutically acceptable salt thereof.
Karbapenemderivater er beskrevet av Choung U. Kim og Peter F. Misco i US-SN 366.910, 471.379 og 389.652. US-SN 366.910 og den derfra avdelte US-SN 471.379, tilsvarende DE-OS 3.312.533, behandler fremstilling av karbapenem-antibiotika med formelen Carbapenem derivatives are described by Choung U. Kim and Peter F. Misco in US-SN 366,910, 471,379 and 389,652. US-SN 366,910 and its assigned US-SN 471,379, corresponding to DE-OS 3,312,533, deal with the preparation of carbapenem antibiotics of the formula
der R<**> betyr hydrogen og R<*> er valgt blant hydrogen; substituert og usubstituert alkyl, alkenyl og alkynyl med fra 1-10 karbonatomer; cykloalkyl og cykloalkylalkyl med 3-6 karbonatomer i cykloalkylringen og 1-6 karbonatomer i alkyldelen; fenyl; aralkyl, aralkenyl og aralkynyl, hvori aryldelen er fenyl; aralkyl, aralkenyl og aralkynyl, hvori aryldelen er fenyl og den alifatiske del har 1-6 karbonatomer; heteroaryl, heteroaralkyl, heterocyklyl og heterocyklylalkyl, hvori heteroatomet eller —atomene i den ovenfor nevnte heterocykliske del er valgt blant 1-4 oksygen-, nitrogen— eller svovelatomer og alkyldelene som er forbundet med de heterocykliske deler har 1-6 karbonatomer; der substituenten eller substituentene med henblikk på de ovenfor nevnte grupper er valgt blant C^_^-alkyl som eventuelt er substituert med amino, halogen, hydroksy eller karboksyl, der med henblikk på de ovenfor nevnte substituenter, gruppene R<3> og R<4> uavhengig er valgt blant hydrogen; alkyl, alkenyl og alkynyl med fra 1-10 karbonatomer; cykloalkyl, cykloalkylalkyl og alkylcykloalkyl med 3-6 karbonatomer i cykloalkylringen og 1-6 karbonatomer i alkyldelen; fenyl; aralkyl, aralkenyl og aralkynyl, hvori aryldelen er fenyl og den alifatlske del har 1-6 karbonatomer; og heteroaryl, heteroaralkyl, heterocyklyl og heterocyklylalkyl, hvori heteroatomet eller -atomene i de ovenfor nevnte heterocykliske deler er valgt blant 1-4 oksygen-, nitrogen- eller svovelatomer og alkyldelene i forbindelse med disse heterocykliske deler har 1-6 karbonatomer eller R<3> og R<4> sammen med nitrogenatomet hvortil minst den ene er bundet, danner en 5-eller 6-leddet nitrogenholdig heterocyklisk ring, R** er som definert for R<3> bortsett fra at den ikke kan være hydrogen; eller der R<1> og R^ sammen danner Cg_^Q<-a>lkyliden eller med hydroksy substituert C210-alkyliden, A er cyklopentylen, cykloheksylen eller C2_^-alkylen, eventuelt substituert med en eller flere C^^-alkylgrupper; R<2> er hydrogen, en anionisk ladning eller en konvensjonell lett fjernbar karboksylbeskyttende gruppe under den forutsetning at det, når R<2> er hydrogen eller en beskyttende gruppe, også finnes et motvirkende anion; og betyr en substituert eller usubstituert mono-, bi- eller polycyklisk aromatisk heterocyklisk gruppe inneholdende minst et nitrogenatom i ringen og bundet til A gjennom et ringnitrogenatom, hvorved det dannes en kvaternær ammoniumgruppe; og farmasøytisk akseptable salter derav ved den i det efterfølgende reaksjonsskjema viste fremgangsmåte: where R<**> means hydrogen and R<*> is selected from hydrogen; substituted and unsubstituted alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl and cycloalkylalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl part; phenyl; aralkyl, aralkenyl and aralkynyl, wherein the aryl moiety is phenyl; aralkyl, aralkenyl and aralkynyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic part are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl parts connected to the heterocyclic parts have 1-6 carbon atoms; where the substituent or substituents with regard to the above-mentioned groups are selected from C^_^-alkyl which is optionally substituted with amino, halogen, hydroxy or carboxyl, where with regard to the above-mentioned substituents, the groups R<3> and R< 4> independently is selected from hydrogen; alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl part; phenyl; aralkyl, aralkenyl and aralkynyl, wherein the aryl moiety is phenyl and the aliphatic moiety has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl parts in connection with these heterocyclic parts have 1-6 carbon atoms or R<3 > and R<4> together with the nitrogen atom to which at least one is bound, form a 5- or 6-membered nitrogen-containing heterocyclic ring, R** is as defined for R<3> except that it cannot be hydrogen; or where R<1> and R^ together form the Cg_^Q<-a>alkylidene or C210-alkylidene substituted with hydroxy, A is cyclopentylene, cyclohexylene or C2_^-alkylene, optionally substituted with one or more C^^-alkyl groups; R<2> is hydrogen, an anionic charge or a conventional readily removable carboxyl protecting group with the proviso that, when R<2> is hydrogen or a protecting group, a counter anion is also present; and means a substituted or unsubstituted mono-, bi- or polycyclic aromatic heterocyclic group containing at least one nitrogen atom in the ring and bonded to A through a ring nitrogen atom, whereby a quaternary ammonium group is formed; and pharmaceutically acceptable salts thereof by the method shown in the following reaction scheme:
US-SN 389.652 og den derfra avledede US-SN 499.690 behandler fremstilling av karbapenem-antibiotika med formelen: US-SN 389,652 and the derivative US-SN 499,690 deal with the preparation of carbapenem antibiotics of the formula:
der R 8 betyr hydrogen og R<*> er valgt blant hydrogen; where R 8 means hydrogen and R<*> is selected from hydrogen;
substituert og usubstituert alkyl, alkenyl og alkynyl med fra 1-10 karbonatomer; cykloalkyl og cykloalkylalkyl med 3-6 karbonatomer i cykloalkylringen og 1-6 karbonatomer i alkyldelen; fenyl; aralkyl, aralkenyl og aralkynyl, hvori aryldelen er fenyl og den alifatiske del har 1-6 karbon- substituted and unsubstituted alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl and cycloalkylalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl part; phenyl; aralkyl, aralkenyl and aralkynyl, in which the aryl part is phenyl and the aliphatic part has 1-6 carbon
atomer; heteroaryl, heteroaralkyl, heterocyklyl og heterocyklylalkyl, hvori heteroatomet eller -atomene i den ovenfor nevnte heterocykliske del er valgt blant 1-4 oksygen-, nitrogen- eller svovelatomer og alkyldelene som er forbundet med de heterocykliske deler har 1-6 karbonatomer, der substituenten eller substituentene med henblikk på de ovenfor nevnte grupper uavhengig er valgt blant C^_^-alkyl som eventuelt er substituert med amino, halogen, hydroksy eller karboksyl, atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, in which the heteroatom or atoms in the above-mentioned heterocyclic part are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl parts connected to the heterocyclic parts have 1-6 carbon atoms, where the substituent or the substituents with regard to the above-mentioned groups are independently selected from C^_^-alkyl which is optionally substituted with amino, halogen, hydroxy or carboxyl,
der med henblikk på de ovenfor nevnte substituenter, gruppene R<3> og R<4> uavhengig er valgt blant hydrogen; alkyl, alkenyl og alkynyl med fra 1-10 karbonatomer; cykloalkyl, cykloalkylalkyl og alkylcykloalkyl med 3-6 karbonatomer i cykloalkylringen og 1-6 karbonatomer i alkyldelen; fenyl; aralkyl, aralkenyl og aralkynyl, der aryldelen er fenyl og den alifatlske del har 1-6 karbonatomer; og heteroaryl, heteroaralkyl, heterocyklyl og heterocyklylalkyl, der heteroatomet eller —atomene i de ovenfor nevnte heterocykliske deler er valgt blant 1-4 oksygen-, nitrogen- eller svovelatomer og where, with respect to the above-mentioned substituents, the groups R<3> and R<4> are independently selected from hydrogen; alkyl, alkenyl and alkynyl having from 1 to 10 carbon atoms; cycloalkyl, cycloalkylalkyl and alkylcycloalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl part; phenyl; aralkyl, aralkenyl and aralkynyl, where the aryl part is phenyl and the aliphatic part has 1-6 carbon atoms; and heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, where the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from 1-4 oxygen, nitrogen or sulfur atoms and
alkyldelene i forbindelse med disse heterocykliske deler har 1-6 karbonatomer eller R<3> og R<4> sammen med nitrogenatomet hvortil minst den ene er bundet, danner en 5- eller 6-leddet, nitrogenholdig, heterocyklisk ring, R 9 er som definert for Rd bortsett fra at den ikke kan være hydrogen; eller der R<*> og R<8> sammen danner Cg^Q-alkyliden eller med hydroksy substituert C210-alkyliden, R<5> er valgt blant substituert og usubstituert alkyl, alkenyl og alkynyl med fra 1-10 karbonatomer; cykloalkyl og cykloalkylalkyl med 3-6 karbonatomer i cykloalkylringen og 1-6 karbonatomer i alkyldelene; fenyl; aralkyl, aralkenyl og aralkynyl, der aryldelen er fenyl og den alifatiske del har 1-6 karbonatomer; heteroaryl, heteroaralkyl, heterocyklyl og heterocyklylalkyl, der heteroatomet eller -atomene i de ovenfor nevnte heterocykliske deler er valgt blant 1-4 oksygen-, nitrogen- eller svovelatomer og alkyldelene som er forbundet med de heterocykliske ringer har 1-6 karbonatomer; der de ovenfor nevnte R<5->grupper eventuelt er substituert med 1-3 substituenter valgt blant: C^^-alkyl, som eventuelt er substituert med amino, fluor, klor, karboksyl, hydroksy eller karbamoyl; fluor, klor eller brom; fenyl, som eventuelt er substituert med 1-3 fluor, klor, brom, C^-alkyl, -OR3, -NR3R4, -S03R<3>, -C02R<3> eller -C0NR<3>R<4>, der R<3>, R<4> og R<9> i disse R<5->substituenter er som definert ovenfor; eller R<5> kan bety en divalent fenylen-eller C1_4-alkylengruppe som er forbundet til ringen under dannelse av en brodannet polycyklisk gruppe; A er cyklopentylen, cykloheksylen eller Cg^-alkylen, som eventuelt er substituert med en eller flere C^^-alkyl-grupper; R<2> er hydrogen, en anionisk ladning eller en konvensjonell lett fjernbar karboksylbeskyttende gruppe under den forutsetning at det, når R<2> er hydrogen eller en beskyttende gruppe, også finnes et motvirkende ion; og betyr en substituert eller usubstituert mono-, bi- eller polycyklisk, ikke-aromatisk, heterocyklisk gruppe inneholdende minst et nitrogenatom i ringen og bundet til A gjennom et ringnitrogenatom, hvorved det dannes en kvaternær ammoniumgruppe; og farmasøytisk akseptable salter derav, ved den i det følgende reaksjonsskjerna viste fremgangsmåte: the alkyl parts in connection with these heterocyclic parts have 1-6 carbon atoms or R<3> and R<4> together with the nitrogen atom to which at least one is bound, form a 5- or 6-membered, nitrogen-containing, heterocyclic ring, R 9 is as defined for Rd except that it cannot be hydrogen; or where R<*> and R<8> together form the C 8-Q alkylidene or hydroxy-substituted C 210 alkylidene, R<5> is selected from substituted and unsubstituted alkyl, alkenyl and alkynyl with from 1-10 carbon atoms; cycloalkyl and cycloalkylalkyl with 3-6 carbon atoms in the cycloalkyl ring and 1-6 carbon atoms in the alkyl moieties; phenyl; aralkyl, aralkenyl and aralkynyl, where the aryl part is phenyl and the aliphatic part has 1-6 carbon atoms; heteroaryl, heteroaralkyl, heterocyclyl and heterocyclylalkyl, where the heteroatom or atoms in the above-mentioned heterocyclic parts are selected from 1-4 oxygen, nitrogen or sulfur atoms and the alkyl parts connected to the heterocyclic rings have 1-6 carbon atoms; where the above-mentioned R<5-> groups are optionally substituted with 1-3 substituents chosen from: C^^-alkyl, which is optionally substituted with amino, fluorine, chlorine, carboxyl, hydroxy or carbamoyl; fluorine, chlorine or bromine; phenyl, which is optionally substituted with 1-3 fluorine, chlorine, bromine, C₁-alkyl, -OR3, -NR3R4, -SO3R<3>, -CO2R<3> or -CONR<3>R<4>, where R<3>, R<4> and R<9> in these R<5->substituents are as defined above; or R<5> can mean a divalent phenylene or C1-4 alkylene group which is attached to the ring forming a bridged polycyclic group; A is cyclopentylene, cyclohexylene or C 3 -alkylene, which is optionally substituted with one or more C 3 -alkyl groups; R<2> is hydrogen, an anionic charge or a conventional easily removable carboxyl protecting group with the proviso that, when R<2> is hydrogen or a protecting group, a counterion is also present; and means a substituted or unsubstituted mono-, bi- or polycyclic, non-aromatic, heterocyclic group containing at least one nitrogen atom in the ring and bonded to A through a ring nitrogen atom, thereby forming a quaternary ammonium group; and pharmaceutically acceptable salts thereof, by the method shown in the following reaction core:
Mere spesielt omsettes utgangsmaterialet III i et inert organisk oppløsningsmiddel med difenylklorfosfat i nærvær av en base under dannelse av mellomprodukt IV. Dette omsettes derefter med en merkaptanreagens med formelen Hs-A-OH i et inert organisk oppløsningsmiddel og i nærvær av en base under dannelse av mellomproduktet V. Dette acyleres derefter med metansulfonylklorid i et inert organisk oppløsningsmiddel og i nærvær av en base under dannelse av mellomproduktet VI som omsettes med en Jodid-ionkilde i et inert organisk oppløs-ningsmiddel under dannelse av mellomproduktet II. Dette omsettes så med det ønskede amin 1 et inert organisk oppløsningsmiddel og i nærvær av et sølvion under dannelse av det kvaterniserte produkt I'A eller I'B som derefter kan deblokkeres under dannelse av det tilsvarende deblokkerte karbapenem med formel IA eller IB. More specifically, the starting material III is reacted in an inert organic solvent with diphenylchlorophosphate in the presence of a base to form intermediate IV. This is then reacted with a mercaptan reagent of the formula Hs-A-OH in an inert organic solvent and in the presence of a base to form the intermediate V. This is then acylated with methanesulfonyl chloride in an inert organic solvent and in the presence of a base to form the intermediate VI which is reacted with an iodide ion source in an inert organic solvent to form the intermediate product II. This is then reacted with the desired amine 1 in an inert organic solvent and in the presence of a silver ion to form the quaternized product I'A or I'B which can then be deblocked to form the corresponding deblocked carbapenem of formula IA or IB.
Den ovenfor beskrevne fremgangsmåte har flere ulemper. Således omfatter fremgangsmåten for eksempel flere trinn, hvorav antall med fordel kunne reduseres. Det samlede reaksjonsutbyttet er også relativt lavt og kvaterniserings-trinnet utføres på hele karbapenemforbindelsen. Det ville være ønskelig å ha en ny fremgangsmåte for fremstilling av forbindelsene med formel IA eller IB som (1) omfatter færre trinn, (2) gir et høyere utbytte, (3) tillater at det kvaterniserte amin dannes først og derefter bindes til karbapenemkjernen på et senere stadium av syntesen og (4) kan anvendes for en enklere dannelse av kvaternære aminprodukter med et stort antall aminer, det vil si aminer med sterisk hindring og med lavere pK^-verdier. The method described above has several disadvantages. Thus, the method includes, for example, several steps, the number of which could be advantageously reduced. The overall reaction yield is also relatively low and the quaternization step is performed on the entire carbapenem compound. It would be desirable to have a new process for the preparation of the compounds of formula IA or IB which (1) involves fewer steps, (2) gives a higher yield, (3) allows the quaternized amine to be formed first and then attached to the carbapenem nucleus on a later stage of the synthesis and (4) can be used for a simpler formation of quaternary amine products with a large number of amines, i.e. amines with steric hindrance and with lower pK^ values.
NO-PS 166038 beskriver en ny og forbedret fremgangsmåte for fremstilling av slike karbapenem-derivater. NO-PS 166038 describes a new and improved method for the production of such carbapenem derivatives.
Foreliggende oppfinnelse tilveiebringer som nevnt ovenfor mellomprodukter med formel (VII) som finner anvendelse for fremstilling av karbapenemforbindelsene med formel (I) ifølge NO-PS 166038, forbindelser som er kraftige antibakterielle midler. The present invention provides, as mentioned above, intermediate products of formula (VII) which find use for the production of the carbapenem compounds of formula (I) according to NO-PS 166038, compounds which are powerful antibacterial agents.
Det kvaternære amintiol-mellomprodukt (VII) ifølge oppfinnelsen kan oppnås ved å arbeide ved en temperatur i området The quaternary amine thiol intermediate (VII) according to the invention can be obtained by working at a temperature in the range
-20'C til +100°C, fortrinnsvis i området 50 til 70°C. -20'C to +100°C, preferably in the range 50 to 70°C.
Det kvaternære amintiol-mellomprodukt vil ha et motanion bundet til seg og som vil være bestemt av den spesielle syre som anvendes. Det er selvfølgelig mulig å substituere med et annet motanion på dette tidspunkt ved hjelp av konvensjonelle fremgangsmåter for anvendelse i den efterfølgende omsetning med karbapenem-mellomprodukt (IV). The quaternary aminothiol intermediate will have a counter anion bound to it and that will be determined by the particular acid used. It is of course possible to substitute with another counter anion at this point using conventional methods for use in the subsequent reaction with carbapenem intermediate (IV).
De følgende eksempler skal belyse oppfinnelsen uten å begrense den. The following examples shall illustrate the invention without limiting it.
Eksempel 1 Example 1
Fremstilling av 3- f2-( l- pvridinlum ) etvltlo1- 6ot-- 81-( R)-hvdroksvetvll- 7- okso- l- azabicyklo( 3. 2. 0)- hept- 2- en- 2-karboksvlat Preparation of 3-f2-(l-pvridinlum)etvltlo1-6ot--81-(R)-hydroxyvetvll-7-oxo-1-azabicyclo(3.2.0)-hept-2-ene-2-carboxvlate
A. l-( 2- merkaptoetvl) pvridlnlummetansulfonat A. 1-(2-Mercaptoethyl)pyridinelumethanesulfonate
Til en suspensjon av pyridiniummetansulfonat i pyridin, fremstilt ved dråpevis tilsetning av metansulfonsyre (1,95 ml, 0,03 mol) til pyridin (8,0 ml, 0,099 mol) under avkjøling ble det satt etylensulfid (1,96 ml, 0,033 mol). Den resulterende blanding ble omrørt ved 55<*>C i 16 timer og konsentrert under redusert trykk til en tykk sirup som ble blandet med noen få ml vann. Oppløsningen ble helt på toppen av en kolonne (40 x 16 cm) av p-Bondapak C^g som ble eluert med vann. Lyofilisering av de passende fraksjoner ga en farveløs sirup, 6,5 g ( 9156). To a suspension of pyridinium methanesulfonate in pyridine, prepared by dropwise addition of methanesulfonic acid (1.95 mL, 0.03 mol) to pyridine (8.0 mL, 0.099 mol) under cooling was added ethylene sulfide (1.96 mL, 0.033 mol ). The resulting mixture was stirred at 55<*>C for 16 hours and concentrated under reduced pressure to a thick syrup which was mixed with a few ml of water. The solution was poured onto the top of a column (40 x 16 cm) of p-Bondapak C^g eluted with water. Lyophilization of the appropriate fractions gave a colorless syrup, 6.5 g (9156).
IR (film) vmaks: 2300-2600 (br. SH), 1635 (pyridinium), 1490, IR (film) vmax: 2300-2600 (br. SH), 1635 (pyridinium), 1490,
1200 (sulfonat), 1068, 1060, 1045, 791, 780 cm-<1>; 1200 (sulfonate), 1068, 1060, 1045, 791, 780 cm-<1>;
<1->H-NMR (DMSO-d6) S: 2,32 (3H, s, CH3S03<e>), 2,61, 2,70, 2,73, <1->H-NMR (DMSO-d6) S: 2.32 (3H, s, CH3SO3<e>), 2.61, 2.70, 2.73,
2,82 (1H, B del av A2B-system, SH), 3,07 (2H, m [med D20, 3,08 (2H, t, J-6,5 Hz)], CH2S), 4,76 (2H, t, J-6,5 Hz, CH2N<®>, 8,19 (2H, m, Hm av pyrldinium), 8,6 (1H, m, HO av pyrldinium), 9,08 (2H, dd, J-6,8 Hz, J-1,4 Hz, HO av pyrldinium); 2.82 (1H, B part of A2B system, SH), 3.07 (2H, m [with D2O, 3.08 (2H, t, J-6.5 Hz)], CH2S), 4.76 (2H, t, J-6.5 Hz, CH2N<®>, 8.19 (2H, m, Hm of pyrldinium), 8.6 (1H, m, HO of pyrldinium), 9.08 (2H, dd , J-6.8 Hz, J-1.4 Hz, HO of pyrldinium);
UV (H20) Xmaks: 206 (c5230), 258 (c3760) nm. UV (H20) Xmax: 206 (c5230), 258 (c3760) nm.
Metode A Method A
B. l-( 2- merkaptoetvl) pyridinlumklorld B. 1-(2-mercaptoethyl) pyridine chloride
En vandig oppløsning av uren l-(2-merkaptoetyl)pyridiniummetansulfonat (9,4 g, 0,04 mol) ble helt på toppen av en kolonne (2,5 x 41 cm) av Permutitt S-l C1<G>. Kolonnen ble eluert med vann i en mengde av 0,5 ml pr. minutt og de passende fraksjoner ble forenet og lyofilisert under dannelse av en gulaktig sirup, 7,0 g (100#), som ble anvendt som den var til neste trinn. An aqueous solution of crude 1-(2-mercaptoethyl)pyridinium methanesulfonate (9.4 g, 0.04 mol) was poured onto the top of a column (2.5 x 41 cm) of Permutitt S-1 C1<G>. The column was eluted with water in an amount of 0.5 ml per minute and the appropriate fractions were combined and lyophilized to give a yellowish syrup, 7.0 g (100#), which was used as is for the next step.
1-H-NMK (D20) S: 3,22 (2H, m, CH2S), 4,88 (m, CH2N<®>), 8,18 1-H-NMK (D2O) S: 3.22 (2H, m, CH2S), 4.88 (m, CH2N<®>), 8.18
(2H, m, Hm av pyrldinium), 8,7 (1H, m, Hp av pyrldinium), 9,0 (2H, m, HO av pyrldinium) ppm. (2H, m, Hm of pyrldinium), 8.7 (1H, m, Hp of pyrldinium), 9.0 (2H, m, HO of pyrldinium) ppm.
Metode B Method B
Til en på forhånd avkjølt (isbad) pyridin (5,6 ml, 70 mmol) ble det satt pyridin-hydroklorid (4,05 g, 35 mmol) og etylensulfid (2,1 ml, 35 mmol). Blandingen ble oppvarmet til 65°C og omrørt i 75 minutter for dannelse av et tofase-system. Den lettere fase ble fjernet. Den tilbakeblevne olje ble vasket med 5 x 10 ml eter og pumpet under høyvakuum for dannelse av 90-100 av tittelforbindelsen, som ble anvendt som sådan til neste trinn. To a previously cooled (ice bath) pyridine (5.6 mL, 70 mmol) was added pyridine hydrochloride (4.05 g, 35 mmol) and ethylene sulfide (2.1 mL, 35 mmol). The mixture was heated to 65°C and stirred for 75 minutes to form a two-phase system. The lighter phase was removed. The residual oil was washed with 5 x 10 ml of ether and pumped under high vacuum to give 90-100 of the title compound, which was used as such for the next step.
C. p- nitrobenzvl- 3- r2-( l- pyridinlum) etvltio1- 6a- ri-( R)-hydroksyetyl"! - 7- okso- l- azabicyklo( 3. 2. 0)- hept- 2- en- 2-karboksylatklorid C. p-nitrobenzvl-3-r2-(1-pyridinelum)ethvlthio1-6a-ri-(R)-hydroxyethyl"!-7-oxo-1-azabicyclo(3.2.0)-hept-2-en- 2-carboxylate chloride
En oppløsning av p-nitrobenzyl-6oc-[l-(R)-hydroksyetyl]-3,7-diokso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat (6,09 g, 17,5 mmol) i 20 ml acetonitril ble avkjølt til +5°C under nitrogenatmosfære og suksessivt behandlet med diisopropyletylamin (3,65 ml, 21,0 mmol) og difenylklorfosfat (4,34 ml, 21,0 mmol). Den resulterende blanding ble omrørt i 30 minutter ved 5"C, avkjølt til -5°C og behandlet suksessivt med en oppløsning av uren l-(2-merkaptoetyl)pyridinlumklorid (4,3 g, 24 mmol) i 1,0 ml N,N-dimetylformamid og dråpevis med diisopropyletylamin (3,65 ml, 21,0 mmol). Reaksjonsblandingen ble omrørt ved 0<*>C 1 1 time, avkjølt til -30"C og omrørt i ytterligere 15 minutter. Det faste stoff ble filtrert av og vasket med kald (-30°C) acetonitril, 5,77 g (6556). A solution of p-nitrobenzyl-6oc-[l-(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate (6.09 g, 17, 5 mmol) in 20 ml of acetonitrile was cooled to +5°C under a nitrogen atmosphere and successively treated with diisopropylethylamine (3.65 ml, 21.0 mmol) and diphenylchlorophosphate (4.34 ml, 21.0 mmol). The resulting mixture was stirred for 30 min at 5°C, cooled to -5°C and treated successively with a solution of crude 1-(2-mercaptoethyl)pyridinelum chloride (4.3 g, 24 mmol) in 1.0 mL of N ,N-dimethylformamide and dropwise diisopropylethylamine (3.65 mL, 21.0 mmol). The reaction mixture was stirred at 0<*>C for 1 hr, cooled to -30°C and stirred for an additional 15 min. The solid was filtered off and washed with cold (-30°C) acetonitrile, 5.77 g (6556).
IR (nujol) vmaks: 3300 (OH), 1775 (C-0 av e-laktam), 1690 IR (nujol) vmax: 3300 (OH), 1775 (C-0 of ε-lactam), 1690
(C-0 av pNB-ester), 1630 (pyrldinium), 1605 (fenyl av pNB-ester), 1515 (N02), 1335 (N02) cm-<1>; (C-0 of pNB ester), 1630 (pyrldinium), 1605 (phenyl of pNB ester), 1515 (NO 2 ), 1335 (NO 2 ) cm-<1>;
^H-NMR (DMS0-d6) S: 1,17 (3H, d, J-6,1 Hz, CH3CH0H), 3,2-3,75 (5H, H-4, H-6, CH2S), 3,75-4,5 (2H, H-5, CH3CHOH), 4,92 (2H, brt, J-6,5 Hz, CH2N°), 5,18 (1H, d, J-4,9 Hz, OH), 5,37 (sentrum av ABq, Ja,b"14»2 Hz» CH2 av PNB), 7,69 (2H, d, J-8,7 Hz, Ho av pNB), 8,24 (d, J-8,7 Hz, Hm av pNB), 8,0-8,4 (4H, Hm av pNB, Hm av pyrldinium), 8,66 (1H, m, Hp av pyrldinium), 9,17 (2H, bred, J-5,5 Hz, Ho av pyrldinium). ^H-NMR (DMS0-d6) S: 1.17 (3H, d, J-6.1 Hz, CH3CH0H), 3.2-3.75 (5H, H-4, H-6, CH2S), 3.75-4.5 (2H, H-5, CH3CHOH), 4.92 (2H, brt, J-6.5 Hz, CH2N°), 5.18 (1H, d, J-4.9 Hz , OH), 5.37 (center of ABq, Ja,b"14»2 Hz» CH2 of PNB), 7.69 (2H, d, J-8.7 Hz, Ho of pNB), 8.24 ( d, J-8.7 Hz, Hm of pNB), 8.0-8.4 (4H, Hm of pNB, Hm of pyrldinium), 8.66 (1H, m, Hp of pyrldinium), 9.17 ( 2H, wide, J-5.5 Hz, Ho of pyrldinium).
Filtratet og vaskevæskene ble forenet og fortynnet med 150 ml eter. Overstående væske ble dekantert og gummien oppløst i 40 ml vann inneholdende tilstrekkelig acetonitril til å holde en oppløsning som ble helt på toppen av en 3 x 10 cm kolonne av The filtrate and washings were combined and diluted with 150 ml of ether. The supernatant liquid was decanted and the gum dissolved in 40 ml of water containing sufficient acetonitrile to hold a solution which was poured onto the top of a 3 x 10 cm column of
-Y-Bondapak C^g. Kolonnen ble eluert med 150 ml 1056 acetonitril - 9056 vann og 100 ml 5056 acetonitril - 5056 vann. De passende fraksjoner ble forenet og lyofilisert efter at acetonitrilet var fjernet under vakuum, under dannelse av et gulaktig pulver. NMR bekrefter nærværet av tittelforbindelsen blandet med noe p-nitrobenzyl-3-[2-(l-pyridinium)etyltio]-6a-[1-(R)-hydroksyetyl]-7-pkso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat-difenylfosfat (2:1). Pulveret ble oppløst i en minimal mengde vann og ført gjennom en 1,5 x 21 cm kolonne av Permutitt S-l Cl<e> med vann. Lyofilisering av de passende fraksjoner ga 1,8 g eller 2056 av tittelforbindelsen. D. p- nltrobenzvl- 3- r2-( l- Pvrldlnl\ im) etvltlo1- 6a- ri-( Rl-hvdroksvetvll- 7- okso- l- azablcvklo( 3. 2. 0) hept- 2- en- 2-karboksvlat- difenylfosfat -Y-Bondapak C^g. The column was eluted with 150 ml of 1056 acetonitrile - 9056 water and 100 ml of 5056 acetonitrile - 5056 water. The appropriate fractions were combined and lyophilized after the acetonitrile was removed under vacuum to give a yellowish powder. NMR confirms the presence of the title compound mixed with some p-nitrobenzyl-3-[2-(1-pyridinium)ethylthio]-6a-[1-(R)-hydroxyethyl]-7-pxo-1-azabicyclo(3.2.0)hept -2-ene-2-carboxylate diphenyl phosphate (2:1). The powder was dissolved in a minimal amount of water and passed through a 1.5 x 21 cm column of Permutitt S-1 Cl<e> with water. Lyophilization of the appropriate fractions gave 1.8 g or 2056 of the title compound. D. p- nltrobenzvl- 3- r2-( l- Pvrldlnl\ im) etvltlo1- 6a- ri-( Rl-hvdroksvetvll- 7- oxo- l- azablcvklo( 3. 2. 0) hept- 2- en- 2- carboxyvlate diphenyl phosphate
En oppløsning av p-nitrobenzyl-6oc-[l-(R)-hydroksyetyl]-3,7-diokso-l-azablcyklo(3.2.0)hept-2-en-2-karboksylat (0,174 g, 0,50 mmol) i 2 ml acetonitril ble avkjølt til 0°C under nitrogenatmosfære og behandlet suksessivt med diisopropyletylamin (0,105 ml, 0,60 mmol) og difenylklorfosfat (0,124 ml, 0,60 mmol). Den resulterende oppløsning ble omrørt i 30 minutter ved 0°C og behandlet suksessivt med en oppløsning av l-(2-merkaptoetyl)pyridinium-metansulfonat (0,170 g, 0,72 mmol) i 0,6 ml acetonitril og diisopropyletylamin (0,105 ml, 0,60 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 15 minutter, fortynnet med 7 ml kaldt vann av 0<*>C og helt på toppen av en 1,5 x 6,4 cm kolonne av ji-Bondapak C^g. Kolonnen ble eluert med en blanding av 25-5096 acetonitril i 75-5056 vann. De passende fraksjoner ble forenet og lyofilisert efter at acetonitrilet var fjernet under vakuum under dannelse av et gulaktig pulver, 0,33 g (9256). A solution of p-nitrobenzyl-6oc-[1-(R)-hydroxyethyl]-3,7-dioxo-1-azablcyclo(3.2.0)hept-2-ene-2-carboxylate (0.174 g, 0.50 mmol ) in 2 mL of acetonitrile was cooled to 0°C under a nitrogen atmosphere and treated successively with diisopropylethylamine (0.105 mL, 0.60 mmol) and diphenylchlorophosphate (0.124 mL, 0.60 mmol). The resulting solution was stirred for 30 min at 0°C and treated successively with a solution of 1-(2-mercaptoethyl)pyridinium methanesulfonate (0.170 g, 0.72 mmol) in 0.6 mL of acetonitrile and diisopropylethylamine (0.105 mL, 0.60 mmol). The reaction mixture was stirred at 0°C for 15 minutes, diluted with 7 mL of cold 0<*>C water and poured onto a 1.5 x 6.4 cm column of ji-Bondapak C^g. The column was eluted with a mixture of 25-5096 acetonitrile in 75-5056 water. The appropriate fractions were combined and lyophilized after the acetonitrile was removed under vacuum to give a yellowish powder, 0.33 g (9256).
IR (KBr) vmaks: 3600-300 (OH), 1765 (C-0 av 3-laktam), 1690 IR (KBr) vmax: 3600-300 (OH), 1765 (C-0 of 3-lactam), 1690
(C-0 av pNB-ester), 1625 (pyrldinium), 1585 (fenyl), 1510 (N02), 1330 (N02), 885 (N02) cm-<1>;(C-0 of pNB ester), 1625 (pyrldinium), 1585 (phenyl), 1510 (NO 2 ), 1330 (NO 2 ), 885 (NO 2 ) cm-<1>;
%-NMR (DMS0-d6) S: 1,16 (3H, d, J-6,2 Hz, CH3CH0H), 4,87 %-NMR (DMS0-d6) S: 1.16 (3H, d, J-6.2 Hz, CH3CH0H), 4.87
(2H, brt, J-6,6 Hz, CH2S), 5,37 (sentrum av ABq, Ja>D-4,3 Hz, CH2 av pNB), 6,7-7,5 (fenyl), 7,68 (d, J-8,8 Hz, Ho av pNB), 8,23 (d, J-8,8 Hz, Hm av pNB), 8,0-8,3 (m, Hm av pyrldinium), 8,4-8,8 (1H, Hp av pyrldinium), 9,09 (2H, dd, J-6,7 Hz, J-1,3 Hz, Ho av pyrldinium). (2H, brt, J-6.6 Hz, CH2S), 5.37 (center of ABq, Ja>D-4.3 Hz, CH2 of pNB), 6.7-7.5 (phenyl), 7, 68 (d, J-8.8 Hz, Ho of pNB), 8.23 (d, J-8.8 Hz, Hm of pNB), 8.0-8.3 (m, Hm of pyrldinium), 8 .4-8.8 (1H, Hp of pyrldinium), 9.09 (2H, dd, J-6.7 Hz, J-1.3 Hz, Ho of pyrldinium).
E. 3- f2-( l- pvridlnium) etvltio1- 6a- ri-( R)- hvdroksvetvll- 7-okso- l- azabicvklo( 3. 2. 0) hept- 2- en- 2- karboksvlat E. 3-f2-(1- pvridlnium)ethvlthio1-6a-ri-(R)-hydroxymethyl-7-oxo-1-azabiclo(3.2.0)hept-2-ene-2-carboxylate
Metode A Method A
Til en oppløsning av p-nitrobenzyl-3-[2-(l-pyridinium)etyl-tio]-6a-[l-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0 )hept-2-en-2-karboksylat-difenylfosfat (0,16 g, 0,22 mmol) i 10 ml våt tetrahydrofuran ble det satt 10 ml eter, monobasisk kaliumfosfat-natriumhydroksyd-buffer, pH 7,4, (16 ml, 0,05 M) og 0,16 g 105é palladium-på-trekull. Den resulterende blanding ble hydrogenert under ca. 3,2 kp/cm<2> i 1 time ved 25'C. De to faser ble separert, og den organiske fase ekstrahert med 2 x 3 ml vann. De vandige opplesninger ble forenet, vasket med 2 x 10 ml eter og helt på toppen av en 1,5 x 6,2 cm kolonne av ji-Bondapak C^g efter at sporene av organiske oppløsningsmidler var fjernet under vakuum. Eluering av kolonnen med vann ga efter lyofilisering av de passende fraksjoner et gulaktig pulver, 0,062 g (8456). To a solution of p-nitrobenzyl-3-[2-(l-pyridinium)ethylthio]-6a-[l-(R)-hydroxyethyl]-7-oxo-l-azabicyclo(3.2.0 )hept-2 -ene-2-carboxylate diphenyl phosphate (0.16 g, 0.22 mmol) in 10 ml of wet tetrahydrofuran was added 10 ml of ether, monobasic potassium phosphate-sodium hydroxide buffer, pH 7.4, (16 ml, 0.05 M) and 0.16 g of 105é palladium-on-charcoal. The resulting mixture was hydrogenated under approx. 3.2 kp/cm<2> for 1 hour at 25'C. The two phases were separated, and the organic phase extracted with 2 x 3 ml of water. The aqueous readings were combined, washed with 2 x 10 ml of ether and poured onto the top of a 1.5 x 6.2 cm column of ji-Bondapak C^g after the traces of organic solvents had been removed under vacuum. Elution of the column with water gave, after lyophilization of the appropriate fractions, a yellowish powder, 0.062 g (8456).
IR (KBr) vmaks: 3700-3000 (OH), 1755 (C-0 av p<->laktam), 1630 IR (KBr) vmax: 3700-3000 (OH), 1755 (C-0 of p<->lactam), 1630
(pyrldinium), 1590 (karboksylat) cm"<1>; (pyrldinium), 1590 (carboxylate) cm"<1>;
<1->H-NMR (D20) S: 1,22 (3H, d, J-6,4 Hz, CH3CH0H), 2,92 (d, <1->H-NMR (D2O) S: 1.22 (3H, d, J-6.4 Hz, CH3CH0H), 2.92 (d,
J-9,1 Hz, H-4), 2,97 (d, J-9,1 Hz, H-4), 3,20 (dd, J=2,5 Hz, J-6,1 Hz, H-6), 3,44 (t, J-6,0 Hz, CH2S), 3,93 (dd, J-9,1 Hz, J-2,5 Hz, H-5), 4,82 (t, J-6,0 Hz, CH2N<®>), 8,04 (m, Hm av pyrldinium), 8,5 (m, Hp av pyrldinium), 8,82 (dd, J-3,2 Hz, J-1,1 Hz, Ho av pyrldinium); J-9.1 Hz, H-4), 2.97 (d, J-9.1 Hz, H-4), 3.20 (dd, J=2.5 Hz, J-6.1 Hz, H-6), 3.44 (t, J-6.0 Hz, CH2S), 3.93 (dd, J-9.1 Hz, J-2.5 Hz, H-5), 4.82 ( t, J-6.0 Hz, CH2N<®>), 8.04 (m, Hm of pyrldinium), 8.5 (m, Hp of pyrldinium), 8.82 (dd, J-3.2 Hz, J-1.1 Hz, Ho of pyrldinium);
UV (H20) <X>maks: 259 (c5800), 296 (c7030) nm; UV (H 2 O) <X>max: 259 (c5800), 296 (c7030) nm;
ttø= 13,5 timer (målt ved en konsentrasjon på 10~<4> M i en ttø= 13.5 hours (measured at a concentration of 10~<4> M in a
fosfatbuffer, pH 7,4 ved 36,8°C). phosphate buffer, pH 7.4 at 36.8°C).
Metode B Method B
Til en oppløsning av p-nitrobenzyl-3-[2-(l-pyridinium)etyl-tio]-6oc-[l-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylatklorid (5,77 g, 11,4 mmol) i monobasisk kaliumfosfat-natritunhydroksyd-buffer (170 ml, 0,2 M, pH 7,22) ble det satt 20 ml tetrahydrofuran, 30 ml eter og 5,7 g 1096 palladium-på-trekull. Den resulterende blanding ble hydrogenert ved 22<*>C under ca. 3,2 kp/cm<2> i 1 time og filtrert på en Celite-pute. Puten ble vasket med 2 x 15 ml vann. Filtratet og vaskevæsken ble forenet og fortynnet med 100 ml eter. Den vandige fase ble separert fra, vasket med 3 x 100 ml eter og helt på toppen av en 4,5 x 20 cm kolonne av >i-Bondapak C^g, efter at de organiske oppløsningsmidler var fjernet under vakuum. Eluering av kolonnen med vann, fulgt av en blanding av 10# acetonitril i vann ga efter lyofilisering av de passende fraksjoner 2,48 g eller 6596 av tittelforbindelsen som et gulaktig pulver. De analytiske data var identiske med de data som ble rapportert for forbindelsen fremstilt under metode A. To a solution of p-nitrobenzyl-3-[2-(1-pyridinium)ethyl-thio]-6oc-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2 -ene-2-carboxylate chloride (5.77 g, 11.4 mmol) in monobasic potassium phosphate-sodium hydroxide buffer (170 ml, 0.2 M, pH 7.22) was added 20 ml of tetrahydrofuran, 30 ml of ether and 5 .7 g 1096 palladium-on-charcoal. The resulting mixture was hydrogenated at 22<*>C under approx. 3.2 kp/cm<2> for 1 hour and filtered on a Celite pad. The pad was washed with 2 x 15 ml of water. The filtrate and washings were combined and diluted with 100 ml of ether. The aqueous phase was separated from, washed with 3 x 100 ml of ether and poured onto the top of a 4.5 x 20 cm column of >i-Bondapak C^g, after the organic solvents had been removed under vacuum. Elution of the column with water, followed by a mixture of 10% acetonitrile in water after lyophilization of the appropriate fractions gave 2.48 g or 6596 of the title compound as a yellowish powder. The analytical data were identical to the data reported for the compound prepared under Method A.
Eksempel 2 Example 2
Fremstilling av 3- r2-( 2-( 3. 5- dimetvlPvrldinlum) etvltio- 6a- ri-( R )- hvdroksyetyl1- 7- okso- l- azablcyklo( 3. 2. 0) hept- 2- en- 2-karboksylat Preparation of 3-r2-(2-(3.5-dimethylPvrldinlum)ethvlthio-6a-ri-(R)-hydroxyethyl1-7-oxo-l-azablcyclo(3.2.0)hept-2-ene-2- carboxylate
A. l-( 2- merkaptoetyl)- 3. 5- dimetvlpyridlniummetansulfonat A. 1-(2- mercaptoethyl)-3.5- dimethylpyridlinium methanesulfonate
Til en suspensjon av 3,5-lutidiniummetansulfonat i 3,5-lutidin, fremstilt ved tilsetning av metansulfonsyre (0,65 ml, 0,010 mol) til kald 3,5-lutidin (2,51 ml, 0,022 mol) settes det etylensulfid (0,655 ml, 0,011 mol). Den resulterende blanding omrøres under nitrogenatmosfaere ved 55'C i 24 timer, avkjøles til 23<*>C og fortynnes med 5 ml vann og 5 ml eter. De organiske sjikt separeres og den vandige oppløsning vaskes med 6 x 4 ml eter. Sporene av eter fjernes under vakuum og oppløsningen settes til toppen av en 2,5 x 6,0 cm kolonne av jj-Bondapak C^g. Kolonnen elueres med vann og lyofilisering av de passende fraksjoner ga en f arveløs sirup, 2,4 g (9196). Ethylene sulfide ( 0.655 mL, 0.011 mol). The resulting mixture is stirred under a nitrogen atmosphere at 55°C for 24 hours, cooled to 23<*>C and diluted with 5 ml of water and 5 ml of ether. The organic layers are separated and the aqueous solution is washed with 6 x 4 ml of ether. The traces of ether are removed under vacuum and the solution added to the top of a 2.5 x 6.0 cm column of jj-Bondapak C^g. The column is eluted with water and lyophilization of the appropriate fractions gave a colorless syrup, 2.4 g (9196).
IR (film) vmaks: 2520 (SH), 1628 (pyrldinium), 1600, 1495, IR (film) vmax: 2520 (SH), 1628 (pyrldinium), 1600, 1495,
1325, 1305, 1283, 1200 (sulfonat), 1040, 938, 765,, 680 cm-1;1325, 1305, 1283, 1200 (sulfonate), 1040, 938, 765,, 680 cm -1 ;
^H-NMR (DMS0-d6) S: 2,31 (3H, s, CH3S03<e>), 2,47 (6H, s, CH3^H-NMR (DMS0-d6) S: 2.31 (3H, s, CH3SO3<e>), 2.47 (6H, s, CH3
på pyrldinium), 2,57, 2,66, 2,69, 2,78 (1H, B del av A2B-system, SE), 3,06 (2H, m [med D20 tilsatt (2H, t, J=6,5 Hz)], CH2S), 4,65 (2H, t, J-6,5 Hz, CH2N<®>), 8,34 (1H, s, Hp av pyrldinium), 8,79 (2H, s, Ho av pyrldinium); on pyrldinium), 2.57, 2.66, 2.69, 2.78 (1H, B part of A2B system, SE), 3.06 (2H, m [with D20 added (2H, t, J= 6.5 Hz)], CH2S), 4.65 (2H, t, J-6.5 Hz, CH2N<®>), 8.34 (1H, s, Hp of pyrldinium), 8.79 (2H, s, Ho of pyrldinium);
UV (H20) <X>maks: 271 (c4860) nm; UV (H 2 O) <X>max: 271 (c4860) nm;
Analyse for Ciq<H>i7N03S2<*>0,5H20: Analysis for Ciq<H>i7N03S2<*>0.5H20:
Beregnet: C 44,09 H 6,66 N 5,14 S 23,54 Calculated: C 44.09 H 6.66 N 5.14 S 23.54
Funnet: C 44,26 H 6,49 N 5,17 S 24,18 Found: C 44.26 H 6.49 N 5.17 S 24.18
B. p- nltrobenzyl- 3- r2-( l-( 3. 5- dlmetylpyridinlum) letvltiol- 6a-1" 1-( R)- hydroksyetyl1- 7- okso- l- azabicyklo( 3. 2. 0) hept-2- en- 2- karboksylatdlfenylfosfat B. p- nltrobenzyl- 3- r2-( l-( 3. 5- dlmethylpyridinlum) letvlthiol- 6a-1" 1-( R)- hydroxyethyl1- 7- oxo- l- azabicyclo( 3. 2. 0) hept- 2- en- 2- carboxylate dlphenyl phosphate
Til en kald (0°C) oppløsning av p-nitrobenzyl-6a-[l-(R)-hydroksyetyl]-3,7-diokso-l-azablcyklo(3.2.0 )hept-2-en-2-karboksylat (0,523 g, 1,50 mmol) i 6,0 ml acetonitril, som holdes under nitrogenatmosfære, ble det satt diisopropyletylamin (0,314 ml, 1,8 mmol) fulgt av difenylklorfosfat (0,373 ml, 1,8 mmol). Reaksjonsblandingen ble omrørt i 30 minutter og behandlet med en oppløsning av l-(2-merkaptoetyl)-3,5-dimetylpyridiniummetansulfonat (0,493 g, 1,87 mmol) i 1,9 ml acetonitril, fulgt av diisopropyletylamin (0,314 ml, 1,8 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 1 time, fortynnet med 26 ml kaldt vann av 0'C og holdt på toppen av en 7,0 x 3,5 cm jj-Bondapak C^g-kolonne. Eluering av kolonnen med 25-5056 acetonitril - 75-5056 vannblanding ga efter lyofilisering av de passende fraksjoner 1,01 g eller 9056 av tittelforbindelsen som et gulaktig pulver. To a cold (0°C) solution of p-nitrobenzyl-6α-[l-(R)-hydroxyethyl]-3,7-dioxo-1-azablcyclo(3.2.0 )hept-2-ene-2-carboxylate ( 0.523 g, 1.50 mmol) in 6.0 mL of acetonitrile, maintained under a nitrogen atmosphere, was added diisopropylethylamine (0.314 mL, 1.8 mmol) followed by diphenylchlorophosphate (0.373 mL, 1.8 mmol). The reaction mixture was stirred for 30 min and treated with a solution of 1-(2-mercaptoethyl)-3,5-dimethylpyridinium methanesulfonate (0.493 g, 1.87 mmol) in 1.9 mL of acetonitrile, followed by diisopropylethylamine (0.314 mL, 1, 8 mmol). The reaction mixture was stirred at 0°C for 1 hour, diluted with 26 mL of cold 0°C water, and retained on top of a 7.0 x 3.5 cm jj-Bondapak C^g column. Elution of the column with 25-5056 acetonitrile - 75-5056 water mixture after lyophilization of the appropriate fractions gave 1.01 g or 9056 of the title compound as a yellowish powder.
IR (KBr) vmaks: 3700-3100 (OH), 1778 (C-0 av e-laktam), 1700 IR (KBr) vmax: 3700-3100 (OH), 1778 (C-0 of ε-lactam), 1700
(C-0 av pNB-ester), 1635 (pyrldinium), 1595 (fenyl), 1521 (N02), 1335 (N02), 895 (N02) cm-<1>; (C-0 of pNB ester), 1635 (pyrldinium), 1595 (phenyl), 1521 (NO 2 ), 1335 (NO 2 ), 895 (NO 2 ) cm-<1>;
^H-NMR (DMS0-d6) 5: 1,16 (3H, dm, J-6,1 Hz, CH3CH0H), 2,43 3H-NMR (DMS0-d6) δ: 1.16 (3H, dm, J-6.1 Hz, CH3CH0H), 2.43
(s, CH3 på pyrldinium), 4,75 (2H, m, CH2N<®>), 5,38 (sentrum av ÅBq, Ja>|j-14,3 Hz, CH2 av pNB), 6,6-7,5 (10H, m, fenyl), 7,70 (2H, d, J-8,7 Hz, Ho av pNB), 8,0-8,5 (3H, m, Hp av pyrldinium, Hm av pNB), 8,82 (2H, s, Ho av pyrldinium); (s, CH3 on pyrldinium), 4.75 (2H, m, CH2N<®>), 5.38 (center of ÅBq, Ja>|j-14.3 Hz, CH2 of pNB), 6.6-7 .5 (10H, m, phenyl), 7.70 (2H, d, J-8.7 Hz, Ho of pNB), 8.0-8.5 (3H, m, Hp of pyrldinium, Hm of pNB) , 8.82 (2H, s, Ho of pyrldinium);
TJV (H20) Xmaks: 270 (cll570), 306 (c7343) nm; TJV (H 2 O) Xmax: 270 (cll570), 306 (c7343) nm;
Analyse for C37<H>3g<N>30ios<p*>H20: Analysis for C37<H>3g<N>30ios<p*>H20:
Beregnet: C 58,03 H 5,26 N 5,48 S 4,18 Funnet: C 57,98 H 5,05 N 5,22 S 4,34 C. 3- r2-( l-( 3. 5- dlmetvlPvridinlum) letvltiol- 6a- fl-( R)-hvdroksvetvll- 7- okso- l- azabicyklo( 3. 2. 0) hept- 2- en- 2-karboksvlat Calculated: C 58.03 H 5.26 N 5.48 S 4.18 Found: C 57.98 H 5.05 N 5.22 S 4.34 C. 3- r2-( l-( 3. 5- dlmetvlPvridinlum) letvlthiol- 6a- fl-( R)-hydroxylvvll- 7- oxo- l- azabicyclo( 3. 2. 0) hept- 2- en- 2-carboxvlate
Til en oppløsning av p-nitrobenzyl-3-[2-(l-(3,5-dimetyl-pyridinium))etyltio]-6a-[l-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylatdifenylfosfat (0,600 g, 0,80 mmol) 1 36 ml våt tetrahydrofuran ble det satt eter (36 ml), monobasisk kaliumfosfat-natriumhydroksydbuffer (0,05 M, pH 7,4, 44 ml) samt 0,60 g 10SÉ palladium-på-trekull. Den resulterende blanding hydrogeneres under ca. 3,2 kp/cm<2> ved 23°C 1 1,25 timer. Det organiske sjikt ble separert fra og ekstrahert med 2 x 5 ml buffer. De vandige sjikt ble forenet, filtrert gjennom en Celite-pute, vasket med 40 ml eter, pumpet for å fjerne spor av organiske oppløsningsmidler og helt på toppen av en 2,5 x 10,0 cm p-Bondapak C^g-kolonne. Eluering av kolonnen med vann og lyofilisering av de passende fraksjoner ga 0,186 g tilsvarende 6456 som et gulaktig pulver. To a solution of p-nitrobenzyl-3-[2-(1-(3,5-dimethyl-pyridinium))ethylthio]-6a-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2 .0)hept-2-ene-2-carboxylate diphenyl phosphate (0.600 g, 0.80 mmol) 1 36 mL of wet tetrahydrofuran was added ether (36 mL), monobasic potassium phosphate-sodium hydroxide buffer (0.05 M, pH 7.4, 44 ml) as well as 0.60 g 10SÉ palladium-on-charcoal. The resulting mixture is hydrogenated under approx. 3.2 kp/cm<2> at 23°C 1 1.25 hours. The organic layer was separated from and extracted with 2 x 5 ml of buffer. The aqueous layers were combined, filtered through a pad of Celite, washed with 40 mL of ether, pumped to remove traces of organic solvents, and poured onto a 2.5 x 10.0 cm p-Bondapak C 2 g column. Elution of the column with water and lyophilization of the appropriate fractions gave 0.186 g corresponding to 6456 as a yellowish powder.
IR (KBr) vmaks: 3700-3100 (OH), 1760 (C-0 av e-laktam), 1595 IR (KBr) vmax: 3700-3100 (OH), 1760 (C-0 of ε-lactam), 1595
(karboksylat) cm"<1>; (carboxylate) cm"<1>;
<i>H-NMR (D20) S; 1,21 (3H, d, J-6,3 Hz, CH3CHOH), 2,45 (6H, <i>H-NMR (D 2 O) S; 1.21 (3H, d, J-6.3 Hz, CH3CHOH), 2.45 (6H,
s, CH3 på pyrldinium), 2,81 (d, J-9,2 Hz, H-4), 2,96 (d, J-9,2 Hz, H-4), 3,22 (dd, J-2,6 Hz, J-6,2 Hz, H-6), 3,40 (t, J-6,2 Hz, CH2S), 3,84 (dd, J-9,2 Hz, J-2,6 Hz, H-5), 4,15 (m, CH3CHOH), 4,71 (t, J-6,2 Hz, CH2N<®>), 8,21 (1H, s, Hp av pyrldinium), 8,46 (2H, s, Ho av pyrldinium); s, CH3 on pyrldinium), 2.81 (d, J-9.2 Hz, H-4), 2.96 (d, J-9.2 Hz, H-4), 3.22 (dd, J -2.6 Hz, J-6.2 Hz, H-6), 3.40 (t, J-6.2 Hz, CH2S), 3.84 (dd, J-9.2 Hz, J-2 .6 Hz, H-5), 4.15 (m, CH3CHOH), 4.71 (t, J-6.2 Hz, CH2N<®>), 8.21 (1H, s, Hp of pyrldinium), 8.46 (2H, s, Ho of pyrldinium);
UV (H20) <X>maks: 2,79 (c8345), 296 (c7714) nm; UV (H 2 O) <X>max: 2.79 (c8345), 296 (c7714) nm;
[a]<g3> +40,7- (c 0,53, H20); [a]<g3> +40.7- (c 0.53, H2O);
ti£ - 16,9 timer (målt ved en konsentrasjon på 10~<4> M i fosfatbuffer, pH 7,4, ved 36,8°C). ten£ - 16.9 hours (measured at a concentration of 10~<4> M in phosphate buffer, pH 7.4, at 36.8°C).
Eksempel 3 Example 3
Fremstilling av ( 5R. 6S)- 3- ff2-( 3- hvdroksvmetvlpyrldinio)-etvll tlol - 6- 1" ( R )- l- hvdroksyetvl" l - 7- okso- l- azablcvklo( 3. 2. 0)-hept- 2- en- 2- karboksvlat Preparation of (5R. 6S)-3- ff2-(3-hydroxymethylpyrldinio)-ethvll tlol - 6- 1" ( R )- l- hydroxyethl" l - 7- oxo- l- azablcvclo( 3. 2. 0)- hept-2-ene-2-carboxylate
A. 3- hvdroksymetvl- l-( 2- merkaptoetvl) pyridlnlum- trlfluor-metansulfonat A. 3-Hydroxymethyl-1-(2-mercaptoethyl)pyridyl-trifluoromethanesulfonate
Trifluormetansulfonsyre (1,327 ml, 0,015 mol) settes dråpevis til 3-pyridinmetanol (2,91 ml, 0,030 mol) fulgt av etylensulfid (0,89 ml, 0,015 mol). Den resulterende homogene blanding ble oppvarmet i et oljebad til 50-70°C under N2 i 20 timer. Reaksjonsblåndingen ble samlet i 15 ml H20 og ekstrahert med 5 x 5 ml CE2C12. Den vandige fase ble konsentrert i vakuum og derefter satt til en C^g-reversfase-kolonne. Eluering med H20, fulgt av inndamping av de relevante fraksjoner ga en blekgul olje. Dette materialet ble omkromatografert under dannelse av en nesten farveløs olje. Efter tørking i vakuum over P20s ga dette produktet i en mengde av 4,50 g tilsvarende 9496 som en viskøs olje. Trifluoromethanesulfonic acid (1.327 mL, 0.015 mol) is added dropwise to 3-pyridinemethanol (2.91 mL, 0.030 mol) followed by ethylene sulfide (0.89 mL, 0.015 mol). The resulting homogeneous mixture was heated in an oil bath to 50-70°C under N 2 for 20 hours. The reaction mixture was collected in 15 mL H 2 O and extracted with 5 x 5 mL CE 2 Cl 2 . The aqueous phase was concentrated in vacuo and then added to a C₂g reverse phase column. Elution with H 2 O, followed by evaporation of the relevant fractions gave a pale yellow oil. This material was rechromatographed to give an almost colorless oil. After drying in vacuo over P20s, this product yielded in an amount of 4.50 g equivalent to 9496 as a viscous oil.
IR (film) vmaks: 3450 (s, OH), 2560 (w, SH) cm-<1>; IR (film) vmax: 3450 (s, OH), 2560 (w, SH) cm-<1>;
^H-NMR (d6-aceton) S: 9,10-8,05 (m, 4H, aromatisk), 5,01 (t, ^H-NMR (d6-acetone) S: 9.10-8.05 (m, 4H, aromatic), 5.01 (t,
J-5,5 Hz, 2H, N-CH2), 4,93 (s, 2H, -CH20H), 4,43 (br S, 1H, -OH), 3,43-3,18 (m, 2H, S-CH2), 2,34-2,10 (m, 1H, SH). J-5.5 Hz, 2H, N-CH2), 4.93 (s, 2H, -CH2OH), 4.43 (br S, 1H, -OH), 3.43-3.18 (m, 2H , S-CH 2 ), 2.34-2.10 (m, 1H, SH).
B. p- nltrobenzvl- f5R. 6S)- 3- r2-( 3- hvdroksvmetvlDvrldinio) etvltiol- 6- ri-( R)- hvdroksvetvn- 7- okso- l- azabicvklo( 3. 2. 0 ^-hept- 2- en- 2- karboksylat- difenylfosfat B. p- nltrobenzvl- f5R. 6S)- 3- r2-( 3- hydroxymethylDvrldinio) ethvlthiol- 6- ri-( R)- hydroxymethyl- 7- oxo- l- azabicclo( 3. 2. 0 ^-hept- 2- ene- 2- carboxylate- diphenyl phosphate
Til en oppløsning av p-nitrobenzyl-(5R,6S)-6-[l-(R)-hydroksy-etyl]-3,7-diokso-l-azabicyklo(3.2.0)heptan-2-karboksylat (0,174 g, 0,50 mmol) i 2 ml tørr acetonitril ble det satt diisopropyletylamin (0,096 ml, 0,55 mmol) ved 0°C vinder N2. Difenylklorfosfat (0,114 ml, 0,55 mmol) ble derefter tilsatt dråpevis og reaksjonsblandingen omrørt ved 0'C i 30 minutter. En oppløsning av 3-hydroksymetyl-l-(2-merkaptoetyl)pyridinium-trifluormetansulfonat (0,223 g, 0,70 mmol) i 0,50 ml acetonitril ble derefter tilsatt, fulgt av diisopropylamin (0,122 ml, 0,70 mmol). Efter at reaksjonsblandingen var blitt holdt ved 0°C i 30 minutter, ble den konsentrert i vakuum og den resterende gule gummi oppsamlet i H20 (det ble tilsatt tilstrekkelig acetonitril for å hjelpe oppløsning av gummien). Denne oppløsning ble satt til en C^g-reversfase-kolonne som ble eluert med 15% acetonitril-B^O. Lyofilisering av de relevante fraksjoner ga 0,305 g tilsvarende 81% av produktet som et belgefarvet, fast stoff. To a solution of p-nitrobenzyl-(5R,6S)-6-[l-(R)-hydroxy-ethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-carboxylate (0.174 g , 0.50 mmol) in 2 ml of dry acetonitrile was added diisopropylethylamine (0.096 ml, 0.55 mmol) at 0°C under N2. Diphenylchlorophosphate (0.114 mL, 0.55 mmol) was then added dropwise and the reaction mixture stirred at 0°C for 30 minutes. A solution of 3-hydroxymethyl-1-(2-mercaptoethyl)pyridinium trifluoromethanesulfonate (0.223 g, 0.70 mmol) in 0.50 mL of acetonitrile was then added, followed by diisopropylamine (0.122 mL, 0.70 mmol). After the reaction mixture was kept at 0°C for 30 minutes, it was concentrated in vacuo and the remaining yellow gum collected in H 2 O (sufficient acetonitrile was added to aid dissolution of the gum). This solution was added to a C₂g reverse phase column eluted with 15% acetonitrile-B₂O. Lyophilization of the relevant fractions gave 0.305 g corresponding to 81% of the product as a pod-coloured solid.
IR (KBr) vmaks: 3420 (br, OH), 1775 (p-laktam CO), 1695 IR (KBr) vmax: 3420 (br, OH), 1775 (p-lactam CO), 1695
(-CO2PNB) cm-1;(-CO2PNB) cm-1;
^H-NMR (dfc-aceton) S: 9,44-7,72 (m, 8H, aromatisk), 7,22-6,91 (m, 10H, difenylfosfat), 5,53, 5,27 (ABq, J-14 Hz, 2H, benzylisk), 5,04 (t, J-7,4 Hz, 2H, N-CH2), 4,75 (s, 2H, CH20H), 4,5-3,1 (m, 8H), 1,21 (d, J=»6,3 Hz, 3H, CHMe). ^H-NMR (dfc-acetone) S: 9.44-7.72 (m, 8H, aromatic), 7.22-6.91 (m, 10H, diphenyl phosphate), 5.53, 5.27 (ABq , J-14 Hz, 2H, benzylic), 5.04 (t, J-7.4 Hz, 2H, N-CH2), 4.75 (s, 2H, CH2OH), 4.5-3.1 ( m, 8H), 1.21 (d, J=»6.3 Hz, 3H, CHMe).
C. ( 5R. 6S)- 3- r2-( 3- hvdroksvmetvlIpvrldinlo) etvltio1- 6- f1-( R)-hydroksyetyll- 7- okso- l- azabicyklo( 3. 2. 0) hept- 2- en- 2-karboksvlat C. ( 5R. 6S)- 3- r2-( 3- hydroxymetvlIpvrldinlo) etvlthio1- 6- f1-( R)-hydroxyethyl- 7- oxo- l- azabicyclo( 3. 2. 0) hept- 2- en- 2 - carbox vat
Til en oppløsning av p-nitrobenzyl-(5R,6S)-3-[2-(3-hydroksy-metylpyridinio )etyltio]-6-[l-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0 )hept-2-en-2-karboksylat-difenylfosfat (0,145 g, 0,194 mmol) i 10 ml THF inneholdende 5 dråper H20 ble det satt 6,0 ml fosfatbuffer (0,05 M, pH 7,4), 0,145 g 10* palladium-på-trekull og 10 ml eter. Blandingen ble hydrogenert (Parr) ved ca. 3,2 kp/cm<2> i 1 time og derefter filtrert gjennom en pute av Celite. Filterkaken ble vasket med noe H2O og eter og den vandige fase separert fra og ekstrahert tre ganger med eter. Den vandige oppløsning ble derefter avkjølt ved 0"C og pH-verdien innstilt til 7,0 med pH 7,4-buffer. Efter fjerning av tilbakeblevne flyktige stoffer i vakuum, ble den vandige oppløsning satt til en C^g-reversfase-kolonne som ble eluert med H2O. Lyofilisering av de relevante fraksjoner ga 36 mg tilsvarende 51* av produktet som et lysegult, fast stoff. Ytterligere rensing ved reversfase-HPLC ga 31 mg eller 41* rent produkt som et fast stoff. To a solution of p-nitrobenzyl-(5R,6S)-3-[2-(3-hydroxymethylpyridinio)ethylthio]-6-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2 .0 )hept-2-ene-2-carboxylate diphenyl phosphate (0.145 g, 0.194 mmol) in 10 ml of THF containing 5 drops of H 2 O was added 6.0 ml of phosphate buffer (0.05 M, pH 7.4), 0.145 g 10* palladium-on-charcoal and 10 ml of ether. The mixture was hydrogenated (Parr) at approx. 3.2 kp/cm<2> for 1 hour and then filtered through a pad of Celite. The filter cake was washed with some H2O and ether and the aqueous phase separated from and extracted three times with ether. The aqueous solution was then cooled at 0°C and the pH adjusted to 7.0 with pH 7.4 buffer. After removal of residual volatiles in vacuo, the aqueous solution was applied to a C 2 g reverse phase column which was eluted with H 2 O. Lyophilization of the relevant fractions gave 36 mg corresponding to 51* of the product as a pale yellow solid. Further purification by reverse phase HPLC gave 31 mg or 41* of pure product as a solid.
IR (KBr) vmaks: 3300 (br, OH), 1755 (e-laktam CO), 1590 IR (KBr) vmax: 3300 (br, OH), 1755 (e-lactam CO), 1590
(-C02<G>) cm-<1>; (-C02<G>) cm-<1>;
^H-NMR (D20) S: 8,78-7,94 (m, 4H, aromatisk), 4,83 (t, J-6,0 ^H-NMR (D 2 O) S: 8.78-7.94 (m, 4H, aromatic), 4.83 (t, J-6.0
Hz, 2H, N-CH2), 4,83 (s, 2H, CH20H), 4,16 (d av q, J-J'-6,2 Hz, 1H, H-l'), 3,98 (d, av t, J-9,1 Hz, J'-2,6 Hz, 1H, H-5), 3,75-3,20 (m, 3H), 3,20-2,65 (m, 2H), 1,22 (d, J-6,4 Hz, 3H, CHMe); Hz, 2H, N-CH2), 4.83 (s, 2H, CH2OH), 4.16 (d of q, J-J'-6.2 Hz, 1H, H-l'), 3.98 ( d, of t, J-9.1 Hz, J'-2.6 Hz, 1H, H-5), 3.75-3.20 (m, 3H), 3.20-2.65 (m, 2H), 1.22 (d, J-6.4 Hz, 3H, CHMe);
UV (H20) Xmaks: 294 (c7614), 266 (c6936) nm; UV (H 2 O) Xmax: 294 (c7614), 266 (c6936) nm;
t% (pH 7,4, 36,8°C); 14,0 t. t% (pH 7.4, 36.8°C); 14.0 h.
Eksempel 4 Example 4
Fremstilling av ( 5R. 6S)- 3- r2-( 4- hvdroksvmetvlp, vridinio) etyl-tiol- 6- fl-( R )- hvdroksvetvll- 7- okso- l- azabicvklo( 3. 2. 0 ) hept- 2-en- 2- karboksyIat A. 4- hydroksymetyl- l-( 2- merkaptoetyl) pyridinium- trlfluor-mfit. anaiil f nn at. Preparation of (5R, 6S)-3-r2-(4-hydroxymethyl, vridinio)ethyl-thiol-6- fl-(R)-hydroxymethyl-7-oxo-1-azabicyclo(3.2.0)hept-2 -en- 2- carboxylate A. 4- hydroxymethyl- 1-( 2- mercaptoethyl) pyridinium- trifluoro-mphyt. anaiil f nn that.
Til en oppløsning av 4-pyridinmetanol (1,635 g, 0,015 mol) i 10 ml CH2CI2 ved 0°C under N2 ble det dråpevis satt trifluormetansulfonsyre (1,327 ml, 0,015 mol). En gulbrun olje skilte seg hurtig ut. En ytterligere ekvivalent 4-pyridinmetanol (1,635 g, 0,015 mol) ble satt til denne blanding og oppløs-ningsmidlet ble fjernet under redusert trykk under dannelse av en olje. Til denne olje ble det satt etylensulfid (0,891 ml, 0,015 mol) og den resulterende homogene blanding ble oppvarmet i et oljebad ved ca. 60"C i 3 timer. Derefter ble reaksjonsblandingen samlet i 15 ml H2O, og den vandige oppløsning vasket med 5 x 5 ml CH2CI2. Efter å ha fjernet resterende organisk oppløsningsmiddel under vakuum, ble den vandige oppløsning satt til en Cig-reversfase-kolonne. Eluering med H20 og efterfølgende inndamping av de relevante fraksjoner ga en olje som ble tørket ytterligere i vakuum over P205, hvorved man oppnådde 4,64 g eller 97* av produktet som en farveløs olje. To a solution of 4-pyridinemethanol (1.635 g, 0.015 mol) in 10 ml of CH 2 Cl 2 at 0°C under N 2 was added dropwise trifluoromethanesulfonic acid (1.327 ml, 0.015 mol). A yellow-brown oil quickly separated. An additional equivalent of 4-pyridine methanol (1.635 g, 0.015 mol) was added to this mixture and the solvent was removed under reduced pressure to leave an oil. To this oil was added ethylene sulfide (0.891 ml, 0.015 mol) and the resulting homogeneous mixture was heated in an oil bath at approx. 60°C for 3 hours. Then the reaction mixture was taken up in 15 mL of H2O, and the aqueous solution was washed with 5 x 5 mL of CH2Cl2. After removing residual organic solvent under vacuum, the aqueous solution was applied to a Cig reverse phase column Elution with H 2 O and subsequent evaporation of the relevant fractions gave an oil which was further dried in vacuo over P 2 O 5 to give 4.64 g or 97% of the product as a colorless oil.
IR (film) vmaks: 3455 (s, OH), 2565 (w, SH) cm-<1>; IR (film) vmax: 3455 (s, OH), 2565 (w, SH) cm-<1>;
1-H-NMR (d6-aceton) S: 9,07, 8,18 (ABq, J-6,8 Hz, 4H, 1-H-NMR (d6-acetone) S: 9.07, 8.18 (ABq, J-6.8 Hz, 4H,
aromatisk), 5,03 (s, 2H, CH20H), 4,96 (t, J-6,5 Hz, 2H, N-CH2), 4,09 (br S, 1H, -OH), 3,5-3,1 (m, 2H, S-CH2), 2,25 (brs, 1H, -SH). aromatic), 5.03 (s, 2H, CH2OH), 4.96 (t, J-6.5 Hz, 2H, N-CH2), 4.09 (br S, 1H, -OH), 3.5 -3.1 (m, 2H, S-CH2), 2.25 (brs, 1H, -SH).
B. p- nltrobenzvl-( 5R. 6S)- 3- r2-( 4- hvdroksvmetylpvrldinlo)-etyltlo~ l- 6-[" l-( R )- l- hvdroksyetyll - 7- okso- l- azabicyklo-( 3. 2. 0) hept- 2- en- 2- karboksvlat- difenylfosfat B. p-nitrobenzvl-(5R.6S)-3-r2-(4-hydroxymethylpvrldinlo)-ethyltlo~l-6-["l-(R)-l-hydroxyethyl-7-oxo-l-azabicyclo-(3 . 2. 0) hept- 2- en- 2- carboxyvlate- diphenyl phosphate
Til en oppløsning av p-nitrobenzyl-(5R,6S)-6-[l-(R)-l-hydroksyetyl]-3,7-diokso-l-azabicyklo(3.2.0)heptan-2-karboksylat (0,348 g, 1,0 mmol) i 5 ml tørr acetonitril ved 0°C under N2 ble det dråpevis satt diisopropyletylamin (0,191 ml, 1,1 mmol), fulgt av difenylklorfosfat (0,228 ml, 1,1 mmol). Den resulterende gyldengule oppløsning ble omrørt ved 0°C i 40 minutter. Til denne oppløsning ble det satt en oppløsning av 4-hydroksymetyl-l-(2-merkaptoetyl)pyridinium-trifluormetansulfonat (0,447 g, 1,4 mmol) i 1 ml acetonitril, efterfulgt av diisopropyletylamin (0,191 ml, 1,1 mmol). En rødlig-sort gummi skilte seg ut fra reaksjonsblandingen. Efter 20 minutter ved 0'C ble reaksjonsblandingen filtrert og konsentrert under vakuum. Resten ble samlet i et minimalt volum (1:1) acetonitril:H20 og satt til en Cig-reversfase-kolonne. Eluering med 25* acetonitril-H20 og efterfølgende lyofilisering av de relevante fraksjoner ga 0,353 g tilsvarende 47* av produktet som et kremfarvet, fast stoff. To a solution of p-nitrobenzyl-(5R,6S)-6-[l-(R)-1-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-carboxylate (0.348 g , 1.0 mmol) in 5 ml of dry acetonitrile at 0°C under N 2 was added dropwise diisopropylethylamine (0.191 ml, 1.1 mmol), followed by diphenylchlorophosphate (0.228 ml, 1.1 mmol). The resulting golden yellow solution was stirred at 0°C for 40 minutes. To this solution was added a solution of 4-hydroxymethyl-1-(2-mercaptoethyl)pyridinium trifluoromethanesulfonate (0.447 g, 1.4 mmol) in 1 mL of acetonitrile, followed by diisopropylethylamine (0.191 mL, 1.1 mmol). A reddish-black gum separated from the reaction mixture. After 20 minutes at 0°C, the reaction mixture was filtered and concentrated under vacuum. The residue was collected in a minimal volume of (1:1) acetonitrile:H 2 O and applied to a C 10 reverse phase column. Elution with 25* acetonitrile-H 2 O and subsequent lyophilization of the relevant fractions gave 0.353 g corresponding to 47* of the product as a cream-colored solid.
IR (KBr) vmakg: 3240 (br, OH), 1775 (e-laktam CO), 1695IR (KBr) vmakg: 3240 (br, OH), 1775 (e-lactam CO), 1695
(-CO2PNB) cm"1;(-CO2PNB) cm"1;
^■H-NMR (d^-aceton) S: 9,24-7,84 (m, H, aromatisk), 7,4-6,9 ^■H-NMR (d^-acetone) S: 9.24-7.84 (m, H, aromatic), 7.4-6.9
(m, 10H, difenylfosfat), 5,52, 5,24 (ABq, J-14 Hz, 2H, benzylisk), 5,15-4,80 (m, 4H), 4,45-3,05 (m, 7H), 1,35 (d, J-6,6 Hz, 3H, CHMe). (m, 10H, diphenyl phosphate), 5.52, 5.24 (ABq, J-14 Hz, 2H, benzylic), 5.15-4.80 (m, 4H), 4.45-3.05 (m , 7H), 1.35 (d, J-6.6 Hz, 3H, CHMe).
C. ( 5R. 6S)- 3- r2-( 4- hvdroksvmetvlpvrldinlo) etvltlo1- b- tl-( R)-l- hydroksvetvll- 7- okso- l- azabicvklo( 3. 2. 0) hept- 2- en- 2-karboksvlat C. ( 5R. 6S)- 3- r2-( 4- hydroxymethylpvrldinlo) etvltlo1- b- tl-( R)-l- hydroxysvetlll- 7- oxo- 1- azabicclo( 3. 2. 0) hept- 2- one - 2-carboxylate
En blanding av p-nitrobenzyl-(5R,6S)-3-[2-(4-hydroksymetyl-pyridinio)etyltio]-6-[l-(R)-l-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0 )hept-2-en-2-karboksylat-difenylfosfat (0,348 g, 0,465 mmol) og 10* palladium-på-trekull (0,35 g) i 11 ml fosfatbuffer (0,05 M, pH 7,4), 5 ml THF og 10 ml eter ble hydrogenert ved ca. 3,2 kp/cm<2> i 1,25 timer. Blandingen ble derefter filtrert gjennom en Celite-pute og den vandige fase vasket med eter tre ganger. pH-verdien i den vandige oppløsning ble derefter innstilt til 7,0 under anvendelse av ytterligere pH 7,4-buffer. Efter å ha fjernet de resterende flyktige stoffer under vakuum ble den vandige oppløsning satt til en Cig-reversfase-kolonne. Eluering med 2* acetonitril-H2O og efterfølgende lyofilisering ga et gulbrunt, fast stoff. Dette materialet ble omkromatografert (Cig-reversfase/g2°) hvorved man oppnådde 0,060 g tilsvarende 36* av det ønskede produkt som et lysegult, fast stoff. A mixture of p-nitrobenzyl-(5R,6S)-3-[2-(4-hydroxymethyl-pyridinio)ethylthio]-6-[1-(R)-1-hydroxyethyl]-7-oxo-1-azabicyclo( 3.2.0 )hept-2-ene-2-carboxylate diphenyl phosphate (0.348 g, 0.465 mmol) and 10* palladium-on-charcoal (0.35 g) in 11 ml of phosphate buffer (0.05 M, pH 7.4 ), 5 ml of THF and 10 ml of ether were hydrogenated at approx. 3.2 kp/cm<2> for 1.25 hours. The mixture was then filtered through a Celite pad and the aqueous phase washed with ether three times. The pH of the aqueous solution was then adjusted to 7.0 using additional pH 7.4 buffer. After removing the remaining volatiles under vacuum, the aqueous solution was applied to a Cig reverse phase column. Elution with 2* acetonitrile-H2O and subsequent lyophilization gave a yellow-brown solid. This material was rechromatographed (Cig reverse phase/g2°) whereby 0.060 g corresponding to 36* of the desired product was obtained as a pale yellow solid.
IR (KBr) vmaks: 3400 (br, 0H), 1755 (e-laktam CO), 1590 IR (KBr) vmax: 3400 (br, 0H), 1755 (e-lactam CO), 1590
(-C02<9>) cm"<1>; (-C02<9>) cm"<1>;
^H-NMR (D20) S: 8,73, 7,96 (ABq, J-6,8 Hz, 4H, aromatisk), ^H-NMR (D 2 O) S: 8.73, 7.96 (ABq, J-6.8 Hz, 4H, aromatic),
4,93 (s, 2H, CH20H), 4,77 (t, J-6,0 Hz, 2H, N-CH2), 4,15 (d av q, J-J'-6,3 Hz, 1H, H-l'), 3,96 (d av t, J-9,2 Hz, J'=2,6 Hz, 1H, H-5), 3,65-3,20 (m, 3H), 3,13-2,62 (m, 2H), 1,21 (d, J-6,3 Hz, 3H, CHMe); 4.93 (s, 2H, CH2OH), 4.77 (t, J-6.0 Hz, 2H, N-CH2), 4.15 (d of q, J-J'-6.3 Hz, 1H , H-l'), 3.96 (d of t, J-9.2 Hz, J'=2.6 Hz, 1H, H-5), 3.65-3.20 (m, 3H), 3.13-2.62 (m, 2H), 1.21 (d, J-6.3 Hz, 3H, CHMe);
UV (H20) <X>maks: 295 (c6880), 256 (c5595), 224 (c8111) nm; , UV (H20) <X>max: 295 (c6880), 256 (c5595), 224 (c8111) nm; ,
t% (pH 7,4, 36,8'C): 14,5 t. t% (pH 7.4, 36.8°C): 14.5 h.
Eksempel 5 Example 5
Fremstilling av 3- T2-( l-( 2- metvlpvridinium)) etvltlo1- 6a- Q-( R)- hvdroksvetvl1- 7- okso- l- azablcvklo( 3. 2. 0) hent- 2- en- 2-karboks<y>lat Preparation of 3- T2-( l-( 2- methylpvridinium)) etvltlo1- 6a- Q-( R)- hvdroxvetl1- 7- oxo- l- azablcvclo( 3. 2. 0) hent- 2- en- 2-carbox <y>lazy
A. l-( 2- merkaptoetyl )- 2- metylpyridlnl^ mmift. nnsulfonat A. 1-(2- mercaptoethyl )- 2- methylpyridnl^ mmift. nnsulfonate
Til en suspensjon av 2-metylpyridiniummetansulfonat i 2-metylpyridin, fremstilt ved tilsetning av metansulfonsyre For a suspension of 2-methylpyridinium methanesulfonate in 2-methylpyridine, prepared by the addition of methanesulfonic acid
(0,65 ml, 0,010 mol) til kald 2-metylpyridin (2,17 ml, 0,022 mol) ble det satt etylensulfid (0,655 ml, 0,011 mol). Reaksjonsblandingen ble omrørt under nitrogen ved 55"C i 21 timer, avkjølt til 23<*>C og fortynnet med 5 ml vann. Den vandige oppløsning ble vasket med 6 x 4 ml eter, pumpet for å. fjerne spor av organiske oppløsningsmidler og helt på toppen av en 2,5 x 10,0 cm p-Bondapak C^g-kolonne. Kolonnen ble eluert med vann og lyofilisering av de passende fraksjoner ga 2,13 g tilsvarende 85* av tittelforbindelsen. (0.65 mL, 0.010 mol) to cold 2-methylpyridine (2.17 mL, 0.022 mol) was added ethylene sulfide (0.655 mL, 0.011 mol). The reaction mixture was stirred under nitrogen at 55°C for 21 hours, cooled to 23<*>C and diluted with 5 mL of water. The aqueous solution was washed with 6 x 4 mL of ether, pumped to remove traces of organic solvents and completely on top of a 2.5 x 10.0 cm p-Bondapak C^g column The column was eluted with water and lyophilization of the appropriate fractions yielded 2.13 g corresponding to 85% of the title compound.
IR (film) vmaks: 2520 (SH), 1623 (pyrldinium), 1574, 1512, IR (film) vmax: 2520 (SH), 1623 (pyrldinium), 1574, 1512,
1485, 1412, 1195 (sulfonat), 1038 cm-<1>; 1485, 1412, 1195 (sulfonate), 1038 cm-<1>;
1-H-NMR (DMS0-d6 + D20) S: 2,37 (3H, s, CH3S03<G>), 2,83 (3H, 1-H-NMR (DMS0-d6 + D2O) S: 2.37 (3H, s, CH3SO3<G>), 2.83 (3H,
s, CH3 på pyrldinium), 3,09 (2H, J-6,9 Hz, CH2S),. 4,71 (2H, t, J=6,9 Hz, CH2N<®>), 7,93 (2H, m, Hm av pyrldinium), 8,44 (1H, m, Hp av pyrldinium), 8,89 (1H, m, Ho av pyrldinium); s, CH3 on pyrldinium), 3.09 (2H, J-6.9 Hz, CH2S), . 4.71 (2H, t, J=6.9 Hz, CH2N<®>), 7.93 (2H, m, Hm of pyrldinium), 8.44 (1H, m, Hp of pyrldinium), 8.89 (1H, m, Ho of pyrldinium);
UV (H20) Xmaks: 266 (c3550) nm. UV (H20) Xmax: 266 (c3550) nm.
B. p- nitrobenzyl- 3- r2-( l-( 2- metylPvridlnlum)) etvltio1- 6a- f1-( R)- hvdroksyetyl1- 7- okso- l- azabicyklo( 3. 2. 0) hept- 2- en- 2-karboksvlat- difenvlfosfat B. p-nitrobenzyl-3- r2-(l-(2-methylPvridlnlum))ethvlthio1-6a-f1-(R)- hydroxyethyl1- 7-oxo-l- azabicyclo(3.2.0)hept-2-ene - 2-carboxylate-diphenylphosphate
Til en kald (0°C) oppløsning av p-nitrobenzyl-6a-[l-(R)-hydroksyetyl]-3,7-diokso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat (0,523 g, 1,50 mmol) i 6 ml acetonitril, og som ble holdt under nitrogenatmosfære, ble det satt diisopropyletylamin (0,314 ml, 1,80 mmol), fulgt av difenylklorfosfat (0,373 ml, 1,80 mmol). Reaksjonsblandingen ble omrørt i 30 minutter ved 0'C og behandlet med en oppløsning av l-(2-merkaptoetyl)-2-metylpyridiniummetansulfonat (0,530 g, 2,16 mmol) i 18 ml acetonitril, fulgt av diisopropyletylamin (0,314 ml, 1,8 mmol). Reaksjonsblandingen ble omrørt ved 0'C i 1 time, fortynnet med 26 ml kaldt vann av 0°C og helt på toppen av en 3,5 x 7,0 cm p-Bondapak C^g-kolonne. Éluering av kolonnen med 25* vann ga efter lyofilisering av de passende fraksjoner 1,06 g tilsvarende 96* av tittelforbindelsen som et gullig pulver. To a cold (0°C) solution of p-nitrobenzyl-6α-[l-(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate ( 0.523 g, 1.50 mmol) in 6 mL of acetonitrile, and kept under a nitrogen atmosphere, was added diisopropylethylamine (0.314 mL, 1.80 mmol), followed by diphenylchlorophosphate (0.373 mL, 1.80 mmol). The reaction mixture was stirred for 30 min at 0°C and treated with a solution of 1-(2-mercaptoethyl)-2-methylpyridinium methanesulfonate (0.530 g, 2.16 mmol) in 18 mL of acetonitrile, followed by diisopropylethylamine (0.314 mL, 1, 8 mmol). The reaction mixture was stirred at 0°C for 1 hour, diluted with 26 mL of cold 0°C water and poured onto a 3.5 x 7.0 cm p-Bondapak C2g column. Elution of the column with 25% water gave, after lyophilization of the appropriate fractions, 1.06 g corresponding to 96% of the title compound as a yellowish powder.
IR (KBr) vmaks: 3650-3100 (OH), 1700 (C-0 av e-laktam), 1695 IR (KBr) vmax: 3650-3100 (OH), 1700 (C-0 of ε-lactam), 1695
og 1690 (C-0 av pNB-ester), 1630 (pyrldinium), 1595 (fenyl), 1518 (N02), 1335 (N02), 890 (N02) cm-<1>; and 1690 (C-0 of pNB ester), 1630 (pyrlidinium), 1595 (phenyl), 1518 (NO 2 ), 1335 (NO 2 ), 890 (NO 2 ) cm-<1>;
^H-NMR (DMS0-d6) 5: 1,15 (3H, d, J-6,1 Hz, CH3CH0H), 2,87 3H-NMR (DMS0-d6) δ: 1.15 (3H, d, J-6.1 Hz, CH3CH0H), 2.87
(s, CH3 på pyrldinium), 3,6-4,4 (2H, m, H-5, CH3CHOH), 4,75 (2H, m, CH2N<®>), 5,37 (sentrum av ABq, J-14 Hz, CH2 av pNB), 6,5-7,4 (10H, m, fenyl), 7,70 (2H, d, J=8,8 Hz, Ho av pNB), 8,0 (2H, m, Hm av pyrldinium), 8,24 (2H, d, J-8,8 Hz, Hm av pNB), 8,50 (1H, m, Hp av pyrldinium), 8,95 (1H, brd, J-6,1 Hz, Ho av pyrldinium); (s, CH3 on pyrldinium), 3.6-4.4 (2H, m, H-5, CH3CHOH), 4.75 (2H, m, CH2N<®>), 5.37 (center of ABq, J -14 Hz, CH2 of pNB), 6.5-7.4 (10H, m, phenyl), 7.70 (2H, d, J=8.8 Hz, Ho of pNB), 8.0 (2H, m, Hm of pyrldinium), 8.24 (2H, d, J-8.8 Hz, Hm of pNB), 8.50 (1H, m, Hp of pyrldinium), 8.95 (1H, brd, J- 6.1 Hz, Ho of pyrldinium);
UV (H20) Xmaks: 265 (cll990), 314 (c8020) nm. UV (H20) Xmax: 265 (cll990), 314 (c8020) nm.
C. 3- r2-( l-( 2- metvlpvrldlnlumnetvltio1- 6a- ri-( Rl- hvdroksvetvl"! - 7- okso- l- azabicyklo( 3. 2. 0 ) hept- 2- en- karboksylat C. 3- r2-( 1-( 2- metvlpvrldlnlumnetvltio1- 6a- ri-( R1- hvdroxvetl"! - 7- oxo-1- azabicyclo( 3. 2. 0 ) hept- 2-ene- carboxylate
Til en oppløsning av p-nitrobenzyl-3-[2-(l-(2-metylpyri-dinium) )etyltio] -6a-[l-(R)-hydroksyetyl] -7-okso-l-azabicyklo-(3.2.0)hept-2-en-2-karboksylat-difenylfosfat (0,66 g, 0,90 mmol) i 34 ml våt tetrahydrofuran ble det satt 34 ml eter, monobasisk kaliumfosfat-natriumhydroksyd-buffer (0,15 M, 16,5 ml, pH 7,22) samt 0,66 g 10* palladium-på-trekull. Den resulterende blanding ble hydrogenert under ca. 3,2 kp/cm<2 >ved 23°C i 1,25 timer. Det organiske sjikt ble separert fra og ekstrahert med 2 x 6 ml buffer. De vandige sjikt ble forenet, filtrert gjennom en Celite-pute, vasket med 40 ml eter, pumpet for å fjerne spor av organiske oppløsningsmidler og helt på toppen av en 2,5 x 10 cm ji-Bondapak C^g-kolonne. Eluering av kolonnen med vann og lyofilisering av de passende fraksjoner ga 0,098 g tilsvarende 31* av tittelforbindelsen som et gulaktig pulver. To a solution of p-nitrobenzyl-3-[2-(1-(2-methylpyridinium))ethylthio]-6a-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo-(3.2. 0)hept-2-ene-2-carboxylate-diphenyl phosphate (0.66 g, 0.90 mmol) in 34 ml of wet tetrahydrofuran was added 34 ml of ether, monobasic potassium phosphate-sodium hydroxide buffer (0.15 M, 16, 5 ml, pH 7.22) as well as 0.66 g 10* palladium-on-charcoal. The resulting mixture was hydrogenated under approx. 3.2 kp/cm<2 >at 23°C for 1.25 hours. The organic layer was separated from and extracted with 2 x 6 ml of buffer. The aqueous layers were combined, filtered through a pad of Celite, washed with 40 mL of ether, pumped to remove traces of organic solvents, and poured onto a 2.5 x 10 cm ji-Bondapak C^g column. Elution of the column with water and lyophilization of the appropriate fractions gave 0.098 g corresponding to 31% of the title compound as a yellowish powder.
IR (KBr) vmaks: 3650-3100 (0H), 1755 (C-0 av e-laktam), 1630 IR (KBr) vmax: 3650-3100 (0H), 1755 (C-0 of ε-lactam), 1630
(pyrldinium) 1595 (karboksylat) cm-<1>; (pyrrlidinium) 1595 (carboxylate) cm-<1>;
^H-NMR (D20) S: 1,20 (3H, d, J-6,3 Hz, CH3CHOH), 2,83 (s, ^H-NMR (D 2 O) S: 1.20 (3H, d, J-6.3 Hz, CH 3 CHOH), 2.83 (s,
CH3 på pyrldinium), 2,7-3,1 (5H, H-4, CH3 på pyrldinium), 3,1-3,7 (3H, m, CH2S, H-6), 3,90 (dd, J-9,1 Hz, J-2,6 Hz, H-5), 3,1 (m, CH3CHOH), 4,78 (t, J-6,2 Hz, CH2N<®>), 7,8 (2H, m, Hm av pyrldinium), 8,3 (1H, m, Hp av pyrldinium), 8,65 (1H, m, Ho av pyrldinium); CH3 on pyrldinium), 2.7-3.1 (5H, H-4, CH3 on pyrldinium), 3.1-3.7 (3H, m, CH2S, H-6), 3.90 (dd, J -9.1 Hz, J-2.6 Hz, H-5), 3.1 (m, CH3CHOH), 4.78 (t, J-6.2 Hz, CH2N<®>), 7.8 ( 2H, m, Hm of pyrldinium), 8.3 (1H, m, Hp of pyrldinium), 8.65 (1H, m, Ho of pyrldinium);
UV (H20) <X>maks: 268 (c9350), 296 (c8840) nm; UV (H 2 O) <X>max: 268 (c9350), 296 (c8840) nm;
[cOg<3> +41- (c 0,5, H20); [cOg<3> +41- (c 0.5, H 2 O);
ti^ - 15,0 timer (målt ved en konsentrasjon på IO-<4> M i fosfatbuffer, pH 7,4 ved 36,8'C). ti^ - 15.0 hours (measured at a concentration of 10-<4> M in phosphate buffer, pH 7.4 at 36.8°C).
1 Eksempel 6 1 Example 6
Fremstilling av 3- f2-( l-( 4- metvlPvrldinium) ) etvltio" l- 6a- ri-( R)- hvdroksyetyl1- 7- okso- l- azablcvklo( 3. 2. 0) hept- 2- en- 2-karboks<y>lat Preparation of 3-f2-(1-(4-methylPvrldinium))ethylthio"1-6a-ri-(R)-hydroxyethyl1-7-oxo-1-azablcvclo(3.2.0)hept-2-ene-2 -karboks<y>lat
A. l-( 2- merkaptoetyl)- 4- metylpyrldlniummetansulfonat A. 1-(2-Mercaptoethyl)-4-methylpyrridlinium methanesulfonate
Til en suspensjon av 4-pikoliniummetansulfonat i 4-pikolin, fremstilt ved tilsetning av metansulfonsyre (0,65 ml, 0,010 mol) til 4-pikolin (2,14 ml, 0,022 mol) under avkjøling, settes etylensulfid (0,655 ml, 0,011 mol). Reaksjonsblandingen omrøres under nitrogen ved 55'C i 24 timer, avkjøles til 23<*>C og fortynnes med 5 ml vann og 10 ml eter. Det organiske sjikt separeres fra og det vandige sjikt vaskes med 5 x 5 ml eter og settes til toppen av en 2,5 x 10 cm jj—Bondapak C^g-kolonne efter at spor av eter var fjernet under redusert trykk. Eluering av kolonnen med en blanding av 15* acetonitril og 85* vann ga efter lyofilisering av de passende fraksjoner 2,66 g tilsvarende 100* av en farveløs sirup. To a suspension of 4-picolinium methanesulfonate in 4-picoline, prepared by adding methanesulfonic acid (0.65 mL, 0.010 mol) to 4-picoline (2.14 mL, 0.022 mol) under cooling, is added ethylene sulfide (0.655 mL, 0.011 mol ). The reaction mixture is stirred under nitrogen at 55°C for 24 hours, cooled to 23<*>C and diluted with 5 ml of water and 10 ml of ether. The organic layer is separated from and the aqueous layer is washed with 5 x 5 ml ether and added to the top of a 2.5 x 10 cm jj—Bondapak C^g column after traces of ether had been removed under reduced pressure. Elution of the column with a mixture of 15* acetonitrile and 85* water gave, after lyophilization of the appropriate fractions, 2.66 g corresponding to 100* of a colorless syrup.
IR (film) vmaks: 2500 (SH), 1640 (pyrldinium), 1572, 1520, IR (film) vmax: 2500 (SH), 1640 (pyrldinium), 1572, 1520,
1478, 1200 (sulfonat), 1040, 833 og 768 cm-<1>; 1478, 1200 (sulfonate), 1040, 833 and 768 cm-<1>;
1-H-NMR (DMS0-d6) S: 2,31 (3H, s, CH3S03<e>), 2,62 (s, CH3 på 1-H-NMR (DMS0-d6) S: 2.31 (3H, s, CH3SO3<e>), 2.62 (s, CH3 on
pyrldinium), 2,2-2,9 (4H, SH, CH3 på pyrldinium),, 3,04 (2H, m, CH2S), 4,68 (2H, t, J-6,4 Hz, CH2N<®>), 8,01 (2H, d, J-6,6 Hz, Hm av pyrldinium), 8,89 (2H, d, J-6,6 Hz, Ho av pyrldinium); pyrldinium), 2.2-2.9 (4H, SH, CH3 on pyrldinium),, 3.04 (2H, m, CH2S), 4.68 (2H, t, J-6.4 Hz, CH2N<® >), 8.01 (2H, d, J-6.6 Hz, Hm of pyrldinium), 8.89 (2H, d, J-6.6 Hz, Ho of pyrldinium);
UV (H20) <X>maks: 256 (c4100), 221 (c7544) nm. UV (H20) <X>max: 256 (c4100), 221 (c7544) nm.
B. l-( 2- merkaptoetvl)- 4- metvlpvrldinium- p- toluensulfonat B. 1-(2-mercaptoethyl)-4-methylpropyldinium-p-toluenesulfonate
Til en suspensjon av p-toluensulfonsyre (1,72 g, 0,01 mol) i 6,5 ml benzen ble det satt 4-pikolin (1,17 ml, 0,012 mol). Den resulterende blanding ble omrørt under nitrogen ved 23°C i 30 minutter, behandlet med etylensulfid (0,65 ml, 0,011 mol) og omrørt ved 75'C i 24 timer. Mere etylensulfid (0,65 ml, 0,011 mol) ble tilsatt, og omrøringen fortsatte ved 75"C i ytterligere 24 timer. Reaksjonsblandingen ble avkjølt til 23°C og fortynnet med 5 ml vann og 8 ml eter. Det vandige sjikt ble separert og vasket med 3 x 8 ml eter. Sporene av organiske oppløsningsmidler ble fjernet under vakuum og forbindelsen kromatografert på y-Bondapak C^g med vann som elueringsmiddel, hvorved man oppnådde 2,94 g eller 90* av tittelforbindelsen som en farvelas sirup. To a suspension of p-toluenesulfonic acid (1.72 g, 0.01 mol) in 6.5 ml of benzene was added 4-picoline (1.17 ml, 0.012 mol). The resulting mixture was stirred under nitrogen at 23°C for 30 min, treated with ethylene sulfide (0.65 mL, 0.011 mol) and stirred at 75°C for 24 h. More ethylene sulfide (0.65 mL, 0.011 mol) was added and stirring was continued at 75°C for an additional 24 hours. The reaction mixture was cooled to 23°C and diluted with 5 mL of water and 8 mL of ether. The aqueous layer was separated and washed with 3 x 8 ml of ether. The traces of organic solvents were removed under vacuum and the compound chromatographed on γ-Bondapak C^g with water as eluent to give 2.94 g or 90% of the title compound as a colored syrup.
IR (film) vmaks: 2510 (SH), 1640 (pyrldinium), 1595, 1582, IR (film) vmax: 2510 (SH), 1640 (pyrldinium), 1595, 1582,
1475, 1200 (sulfonat), 1031, 1010, 818 cm-<1>. 1475, 1200 (sulfonate), 1031, 1010, 818 cm-<1>.
^H-NMR (DMS0-d6) S: 2,29 (3H, s, CH3 på pyrldinium), 2,61 ^H-NMR (DMS0-d6) S: 2.29 (3H, s, CH3 on pyrldinium), 2.61
(s, CH3, Ph), 2,4-2,8 (4H, SH, CH3Ph), 3,03 (2H, m [tilsetning av D2O ga en t, J-6,4 Hz, ved 3,04], CH2S), 4,68 (2H, t, J-6,4 Hz, CH2N<®>), 7,11, 7,49 (4H, 2d, J-7,9 Hz, fenyl), 8,00 (2H, d, J-6,5 Hz, Hm av pyrldinium), 8,89 (2H, d, J-6,5 Hz, Ho av pyrldinium); (s, CH3, Ph), 2.4-2.8 (4H, SH, CH3Ph), 3.03 (2H, m [addition of D2O gave a t, J-6.4 Hz, at 3.04] , CH2S), 4.68 (2H, t, J-6.4 Hz, CH2N<®>), 7.11, 7.49 (4H, 2d, J-7.9 Hz, phenyl), 8.00 (2H, d, J-6.5 Hz, Hm of pyrldinium), 8.89 (2H, d, J-6.5 Hz, Ho of pyrldinium);
UV (H20) <X>maks: 256 (c4315), 222 (cl7045) nm. UV (H20) <X>max: 256 (c4315), 222 (cl7045) nm.
C. p- nitrobenzvl- 3- r2-( l-( 4- metylpyridinlum)) etyltlo1- 6a- f1-( R)- hvdroksvetvl1- 7- okso- l- azablcyklo( 3. 2. 0) hept- 2- en- 2-karboksy1at- d ifenylfosfat C. p- nitrobenzvl- 3- r2-( l-( 4- methylpyridinlum)) ethyltlo1- 6a- f1-( R)- hydroxyvetl1- 7- oxo- l- azablcyclo( 3. 2. 0) hept- 2- one - 2-carboxyl-diphenyl phosphate
Til en kald (0<*>C) oppløsning av p-nitrobenzyl-6a-[l-(R)-hydroksyetyl]-3,7-diokso-l-azabicyklo(3.2.0)hept-2-en-karboksylat (0,522 g, 1,5 mmol) i 6 ml acetonitril, holdt under en nitrogenatmosfære, ble det satt diisopropyletylamin (0,314 ml, 1,8 mmol) fulgt av difenylklorfosfat (0,373 ml, 1,9 mmol). Reaksjonsblandingen ble omrørt i 45 minutter og behandlet dråpevis med en oppløsning av l-(2-merkaptoetyl)-4-metylpyridiniummetansulfonat (0,539 g, 2,16 mmol) i 1,8 ml acetonitril, fulgt av diisopropyletylamin (0,314 ml, 1,8 mmol). Reaksjonsblandingen ble omrørt ved 0<*>C i 1 time, fortynnet med 24 ml kaldt vann av 0'C og helt på toppen av en 2,5 x 8,5 cm jj-Bondapak C^g-kolonne. Eluering av kolonnen, først med 100 ml av en blanding av 25* acetonitril og 75* vann og derefter med 100 ml av en blanding av 50* acetonitril og 50* vann, ga efter lyofilisering av de passende fraksjoner 0,91 g eller 83* av tittelforbindelsen som et gulaktig pulver. To a cold (0<*>C) solution of p-nitrobenzyl-6a-[l-(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)hept-2-ene carboxylate ( 0.522 g, 1.5 mmol) in 6 mL of acetonitrile, maintained under a nitrogen atmosphere, was added diisopropylethylamine (0.314 mL, 1.8 mmol) followed by diphenylchlorophosphate (0.373 mL, 1.9 mmol). The reaction mixture was stirred for 45 min and treated dropwise with a solution of 1-(2-mercaptoethyl)-4-methylpyridinium methanesulfonate (0.539 g, 2.16 mmol) in 1.8 mL of acetonitrile, followed by diisopropylethylamine (0.314 mL, 1.8 mmol). The reaction mixture was stirred at 0<*>C for 1 hour, diluted with 24 mL of cold 0°C water and poured onto a 2.5 x 8.5 cm jj-Bondapak C^g column. Elution of the column, first with 100 ml of a mixture of 25* acetonitrile and 75* water and then with 100 ml of a mixture of 50* acetonitrile and 50* water, gave, after lyophilization of the appropriate fractions, 0.91 g or 83* of the title compound as a yellowish powder.
IR (KBr) vmaks: 3700-2800 (OH), 1770 (C-0 av e-laktam), IR (KBr) vmax: 3700-2800 (OH), 1770 (C-0 of ε-lactam),
1700 (C-0 av pNB-ester), 1640 (pyrldinium), 1595 (fenyl), 1520 (N02), 1340 (N02), 890 (N02) cm-<1>; 1700 (C-0 of pNB ester), 1640 (pyrlidinium), 1595 (phenyl), 1520 (NO2), 1340 (NO2), 890 (NO2) cm-<1>;
<i>H-NMR (DMS0-d6) S: 1,16 (3H, d, J-6,2 Hz, CH3CH0H), 2,61 <i>H-NMR (DMS0-d6) S: 1.16 (3H, d, J-6.2 Hz, CH3CH0H), 2.61
(s, CH3 på pyrldinium), 3,1-3,7 (3H, m, H-6, CH2S), 3,7-4,4 (2H, m, H-5, CH3CHOH), 4,79 (2H, brt, J-6,3 Hz, CH2N<®>), 5,17 (d, J-4,9 Hz, OH), 5,37 (sentrum av ABq, J=14,l Hz, CH2 av pNB), 6,7-7,4 (10H, m, fenyl), 7,69 (2H, d, J-8,8 Hz, Ho av pNB), 8,00 (2H, d, J-6,5 Hz, Hm av pyrldinium), 8,23 (2H, d, J-8,8 Hz, Hm av pNB), 8,92 (2H, d, J-6,5 Hz, Ho av pyrldinium); (s, CH3 on pyrldinium), 3.1-3.7 (3H, m, H-6, CH2S), 3.7-4.4 (2H, m, H-5, CH3CHOH), 4.79 ( 2H, brt, J-6.3 Hz, CH2N<®>), 5.17 (d, J-4.9 Hz, OH), 5.37 (center of ABq, J=14.1 Hz, CH2 of pNB), 6.7-7.4 (10H, m, phenyl), 7.69 (2H, d, J-8.8 Hz, Ho of pNB), 8.00 (2H, d, J-6, 5 Hz, Hm of pyrldinium), 8.23 (2H, d, J-8.8 Hz, Hm of pNB), 8.92 (2H, d, J-6.5 Hz, Ho of pyrldinium);
UV (H20) Xmaks: 262 (cl0835), 311 (c9670) nm; UV (H 2 O) Xmax: 262 (cl0835), 311 (c9670) nm;
Analyse for C36H36N3010SP'1,5H20: Analysis for C36H36N3010SP'1.5H20:
Beregnet: C 56,84 H 5,17 N 5,52 S 4,21 Calculated: C 56.84 H 5.17 N 5.52 S 4.21
Funnet: C 56,89 H 5,13 N 5,19 S 4,41 Found: C 56.89 H 5.13 N 5.19 S 4.41
D. 3- r2-( l-( 4- metvlpvrldlniumnetvltlo1- 6a- ri-( R)- hvdroksv-etyll- 7- okso- l- azablcyklo( 3. 2. 0) hept- 2- en- 2- karboksvlat D. 3- r2-( l-( 4- metvlpvrldlniumnetvltlo1- 6a- ri-( R)- hydroxy-ethyl- 7- oxo- l- azablcyclo( 3. 2. 0) hept- 2- en- 2- carboxylate
Til en oppløsning av p-nitrobenzyl-3-[2-(l-(4-metylpyridi-nium))etyltlo]-6oc-[l-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat-difenylfosfat (0,587 g, 0,80 mmol) i 30 ml våt tetrahydrofuran ble det satt 30 ml eter, monobasisk kaliumfosfat-natriumhydroksydbuffer (0,15 M, 14,7 ml, pH 7,22)og 0,59 g 10* palladium-på-trekull. Den resulterende blanding ble hydrogenert under ca. 32 kp/cm<2> ved 23°C i 1,25 timer. Det organiske sjikt ble separert og ekstrahert med 2 x 6 ml buffer. De vandige ekstrakter ble forenet, filtrert gjennom en Celite-pute, vasket med 3 x 20 ml eter, pumpet for fjerning av spor av organiske oppløs-ningsmidler og helt på toppen av en 2,5 x 10 cm kolonne av jj—Bondapak C^g. Eluering av kolonnen med vann og lyofilisering av de passende fraksjoner ga 0,136 g tilsvarende 49* av tittelforbindelsen som et gulaktig pulver. To a solution of p-nitrobenzyl-3-[2-(1-(4-methylpyridinium))ethyltlo]-6oc-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0 )hept-2-ene-2-carboxylate diphenyl phosphate (0.587 g, 0.80 mmol) in 30 mL of wet tetrahydrofuran was added 30 mL of ether, monobasic potassium phosphate-sodium hydroxide buffer (0.15 M, 14.7 mL, pH 7 .22) and 0.59 g of 10* palladium-on-charcoal. The resulting mixture was hydrogenated under approx. 32 kp/cm<2> at 23°C for 1.25 hours. The organic layer was separated and extracted with 2 x 6 ml of buffer. The aqueous extracts were combined, filtered through a pad of Celite, washed with 3 x 20 ml of ether, pumped to remove traces of organic solvents and poured onto a 2.5 x 10 cm column of jj-Bondapak C^ g. Elution of the column with water and lyophilization of the appropriate fractions gave 0.136 g corresponding to 49% of the title compound as a yellowish powder.
IR (KBr) vmaks: 3700-3000 (0H), 1770 (C=0 av e-laktam), 1642 IR (KBr) vmax: 3700-3000 (0H), 1770 (C=0 of ε-lactam), 1642
(pyrldinium), 1592 (karboksylat) cm-<1>; (pyrrlidinium), 1592 (carboxylate) cm-<1>;
^-NMR (D20) S: 1,19 (3H, s, J-6,3 Hz, CH3CHOH), 2,59 (3H, 3-NMR (D 2 O) S: 1.19 (3H, s, J-6.3 Hz, CH 3 CHOH), 2.59 (3H,
s, CH3 på pyrldinium), 2,84 (d, J-9,1 Hz, H-4), 2,90 (d, J-9,1 Hz, H-4), 3,0-3,6 (3H, m, CH2S, H-6), 3,86 (dd, J-9,1 Hz, J-2,6 Hz, H-5), 4,12 (m, CH3CHOH), 4,5-4,9 (CH2N<®> maskert med HOD), 7,80 (2H, d, J-5,6 Hz, Hm av pyrldinium), 8,58 (2H, d, J=6,6 Hz, Ho av pyrldinium); s, CH3 on pyrldinium), 2.84 (d, J-9.1 Hz, H-4), 2.90 (d, J-9.1 Hz, H-4), 3.0-3.6 (3H, m, CH2S, H-6), 3.86 (dd, J-9.1 Hz, J-2.6 Hz, H-5), 4.12 (m, CH3CHOH), 4.5- 4.9 (CH2N<®> masked with HOD), 7.80 (2H, d, J-5.6 Hz, Hm of pyrldinium), 8.58 (2H, d, J=6.6 Hz, Ho of pyrldinium);
UV (H20) Xmaks: 256 (c5510), 262 (c5360), 296 (c7050) nm; UV (H2O) Xmax: 256 (c5510), 262 (c5360), 296 (c7050) nm;
[oc]g3 -20,8' (c 0,48, H20); [oc]g 3 -20.8' (c 0.48, H 2 O);
ty = 12,8 timer (målt ved en konsentrasjon på 10~<4> M i en ty = 12.8 hours (measured at a concentration of 10~<4> M in a
fosfatbuffer, pH 7,4 ved 36,8°C). phosphate buffer, pH 7.4 at 36.8°C).
Eksempel 7 Example 7
Fremstilling av ( 5R)- 3- f2-( 4- metvltiopvridinioletvltiol-( 6S)-fl-( R )- hydroksyetyl1- 7- okso- l- azablcvklo( 3. 2. 0) hept- 2- en- 2-karboks<y>lat Preparation of (5R)-3-f2-(4-methylthiopvridinioletvlthio-(6S)-fl-(R)-hydroxyethyl1-7-oxo-1-azablcvclo(3.2.0)hept-2-ene-2-carbox <y>lazy
A. 4- metyltiopyridin-^<1> Fremstilling av denne forbindelse ble rapportert i King & Ware, "J. Chem. Soc", 873 (1939). Man fulgte den her beskrevne prosedyre. A. 4-Methylthiopyridine-^<1> The preparation of this compound was reported in King & Ware, "J. Chem. Soc", 873 (1939). The procedure described here was followed.
4-merkaptopyridin (5,55 g, 50,0 mmol; Aldrich) ble oppløst i 50 ml kokende absolutt EtOH. Det uoppløselige materialet ble fjernet ved filtrering over Celite. Filtratet ble oppvarmet til gjenoppløsning og efter avkjøling til ca. 50°C ble det på en gang tilsatt metyljodid (3,17 ml, 51,0 mmol; Aldrich). Blandingen ble avkjølt til krystallisering. Filtrering av det faste stoff ga 6,77 g (26,7 mmol, utbytte 53,5*) av tittelforbindelsen som hydrojodid. 4-Mercaptopyridine (5.55 g, 50.0 mmol; Aldrich) was dissolved in 50 mL of boiling absolute EtOH. The insoluble material was removed by filtration over Celite. The filtrate was heated to redissolve and after cooling to approx. At 50°C, methyl iodide (3.17 mL, 51.0 mmol; Aldrich) was added at once. The mixture was cooled to crystallization. Filtration of the solid gave 6.77 g (26.7 mmol, yield 53.5*) of the title compound as hydroiodide.
^H-NMR (D20) S: 2,70 (3H, s, -SCH3) og 7,65-7,77-8,35-8,48 ^H-NMR (D 2 O) S: 2.70 (3H, s, -SCH 3 ) and 7.65-7.77-8.35-8.48
(4H, A2B2-type, aromatisk-Hs) ppm; (4H, A2B2 type, aromatic-Hs) ppm;
IR (nujol) vmaks: 1615, 1585 (aromatisk) og 780 cm"<1>; IR (nujol) vmax: 1615, 1585 (aromatic) and 780 cm"<1>;
UV (H20) Xmaks: 227 (c2,20 x IO<4>) og 298 (cl,64 x 10<4>) nm.1 UV (H20) Xmax: 227 (c2.20 x IO<4>) and 298 (cl.64 x 10<4>) nm.1
Hydrojodidet (6,33 g, 25,0 mmol) ble oppløst i 40 ml H20 og det uoppløselige materialet fjernet og vasket med 10 ml H20. Til filtratet ble det ved 0-5"C satt 5 g NaOH-tabletter og man ekstraherte med 3 x 25 ml Et20, det vandige sjikt ble mettet med NaCl. De forenede organiske ekstrakter ble vasket to ganger med saltvann, tørket over MgS04 og inndampet til et utbytte på 2,92 g (23,4 mmol, totalt utbytte 50*) av tittelforbindelsen som en olje; The hydroiodide (6.33 g, 25.0 mmol) was dissolved in 40 mL H 2 O and the insoluble material removed and washed with 10 mL H 2 O. 5 g NaOH tablets were added to the filtrate at 0-5"C and extracted with 3 x 25 ml Et2O, the aqueous layer was saturated with NaCl. The combined organic extracts were washed twice with brine, dried over MgSO4 and evaporated to yield 2.92 g (23.4 mmol, total yield 50*) of the title compound as an oil;
<i>H-NMR (CDC13) S: 2,48 (3H, s, -SCH3) og 7,03-7,13-8,38-8,48 <i>H-NMR (CDCl 3 ) S: 2.48 (3H, s, -SCH 3 ) and 7.03-7.13-8.38-8.48
(4H, A2B2-type, aromatisk-Hs) ppm; (4H, A2B2 type, aromatic-Hs) ppm;
IR (film) vmaks: 1580 og 800 cm-<1.>IR (film) vmax: 1580 and 800 cm-<1.>
B. 4- metvltio- N-( 2- merkaptoetvl) pvridinlummetansulfonat B. 4-Methylthio-N-(2-mercaptoethyl)pyridinelumethanesulfonate
4-metyltiopyridin (2,75 g, 22,0 mmol) ble satt langsomt til metansulfonsyre<2> (0,65 ml, 10,5 mmol) under avkjøling i et isbad. Til dette faste stoff ble det satt etylensulfid2 (0,66 ml, 11,0 mmol, Aldrich), og blandingen ble oppvarmet til 50-60°C i 2 timer. Eftersom reaksjonen skred frem, gikk det faste stoff i oppløsning. Efter avkjøling ble reaksjonsblandingen oppløst i 5 ml H2O og vasket med 5 x 4 ml Et20. Det uklare vandige sjikt ble filtrert over Celite og filtratet renset ved reversfase-silikagel-kolonnekromatografi (Cig-y-Bondapak, 10 g) og man eluerte med H2O. Hver fraksjon på 10 ml ble samlet. Fraksjonene 2 og 3 ble forenet og renset igjen ved hjelp av reversfase-kolonnen. Fraksjon 2 ga 1,258 g (4,48 mmol, utbytte 42,6*) av tittelforbindelsen som en viskøs olje. 4-Methylthiopyridine (2.75 g, 22.0 mmol) was added slowly to methanesulfonic acid<2> (0.65 mL, 10.5 mmol) while cooling in an ice bath. To this solid was added ethylene sulfide 2 (0.66 mL, 11.0 mmol, Aldrich), and the mixture was heated to 50-60°C for 2 hours. As the reaction progressed, the solid dissolved. After cooling, the reaction mixture was dissolved in 5 ml H2O and washed with 5 x 4 ml Et2O. The cloudy aqueous layer was filtered over Celite and the filtrate purified by reverse phase silica gel column chromatography (Cig-y-Bondapak, 10 g) and eluted with H 2 O. Each fraction of 10 ml was pooled. Fractions 2 and 3 were combined and purified again using the reverse phase column. Fraction 2 gave 1.258 g (4.48 mmol, yield 42.6*) of the title compound as a viscous oil.
<1->H-NMR (DMS0-d6, CFT-20) S: 2,32 (3H, s, MeS03<e>), 2,72 (3H, <1->H-NMR (DMS0-d6, CFT-20) S: 2.32 (3H, s, MeSO3<e>), 2.72 (3H,
s, -SMe), 2,68 (1H, m, SH), 2,9-3,2 (3H, m, -H2S-), 4,59 (2H, t, J=6,4 Hz, -CH2N<®>), 7,97 (2H, "d", J-7,2 Hz, aromatisk-Hs) og 8,72 (2H, "d", J-7,2 Hz, aromatisk-Hz) ppm; s, -SMe), 2.68 (1H, m, SH), 2.9-3.2 (3H, m, -H2S-), 4.59 (2H, t, J=6.4 Hz, - CH2N<®>), 7.97 (2H, "d", J-7.2 Hz, aromatic-Hs) and 8.72 (2H, "d", J-7.2 Hz, aromatic-Hz) ppm ;
IR (ren) vmaks: 1630, 1200 (br, -S03<G>), <7>85 og 770 cm-<1>. IR (pure) vmax: 1630, 1200 (br, -S03<G>), <7>85 and 770 cm-<1>.
C. ( 5R)- p- nltrobenzvl- 3- r2-( 4- metvltiopvridlnio) etvltio1-( 6S)- f1-( R)- hvdroksvetvl1- 7- okso- l- azabicvklo( 3. 2. 0 ) hept-2- en- 2- karboksvlatklorid C. (5R)- p- niltrobenzvl- 3- r2-( 4- metvlthiopvridlnio) etvlthio1-( 6S)- f1-( R)- hvdroxvevl1- 7- oxo- l- azabicclo( 3. 2. 0 ) hept-2 - en- 2- carboxyl chloride
<2> Disse reagenser destilleres før bruk. <2> These reagents are distilled before use.
Til en oppløsning av (5R)-p-nitrobenzyl-3,7-diokso-(6S)-[(IR)-hydroksyetyl]-l-azabicyklo(3.2.0)heptan-(2R)-karboksylat (475 mg, 1,36 mmol) og diisopropyletylamin (0,24 ml, 1,4 mmol) i CH3CN (5 ml) ble det ved 0-5'C under nitrogen satt difenylklorfosfat (0,29 ml, 1,41 mmol). Blandingen ble omrørt ved 0-5°C i 30 minutter. Til denne blanding ble det satt en oljeaktig suspensjon av 4-metyltio-N-(2-merkaptoetyl)-pyridiniummetansulfonat (678 mg, 1,45 mmol; 60* rent) i 1,5 ml CH3CN, fulgt av diisopropyletylamin (0,24 ml, 1,4 mmol). Blandingen ble omrørt ved 0-5°C i 1 time. Umiddelbart efter tilsetning av basen ble det dannet et gulaktig bunnfall. Dette ble filtrert av og vasket med 3 ml kald CE3CN til et utbytte på 314 mg av et gulaktig, fast stoff. Dette ble triturert fra 5 ml 10* MeOE i H2O og man oppnådde 341 mg (0,618 mmol, utbytte 45,4*) av tittelforbindelsen i form av hvite krystaller med smeltepunkt 118-120°C. To a solution of (5R)-p-nitrobenzyl-3,7-dioxo-(6S)-[(IR)-hydroxyethyl]-1-azabicyclo(3.2.0)heptane-(2R)-carboxylate (475 mg, 1 .36 mmol) and diisopropylethylamine (0.24 ml, 1.4 mmol) in CH3CN (5 ml) at 0-5°C under nitrogen was added diphenylchlorophosphate (0.29 ml, 1.41 mmol). The mixture was stirred at 0-5°C for 30 minutes. To this mixture was added an oily suspension of 4-methylthio-N-(2-mercaptoethyl)-pyridinium methanesulfonate (678 mg, 1.45 mmol; 60* pure) in 1.5 mL of CH 3 CN, followed by diisopropylethylamine (0.24 ml, 1.4 mmol). The mixture was stirred at 0-5°C for 1 hour. Immediately after addition of the base, a yellowish precipitate was formed. This was filtered off and washed with 3 mL of cold CE 3 CN to yield 314 mg of a yellowish solid. This was triturated from 5 ml of 10* MeOE in H2O and 341 mg (0.618 mmol, yield 45.4*) of the title compound was obtained in the form of white crystals with a melting point of 118-120°C.
1-H-NMR (DMS0-d6, CFT-20) S: 1,16 (3H, d, J-6,1 Hz, 1<*->CH3), 1-H-NMR (DMS0-d6, CFT-20) S: 1.16 (3H, d, J-6.1 Hz, 1<*->CH3),
2,72 (3H, s, -CH3), 3,1-3,7 (5H, m), 3,7-4,3 (2H, m), 4,71 (2H, t, J-6,3 Hz, -CH2N<®>), 5,15 (1H, d, J-4,9 Hz, OH), 5,20-5,35-5,40-5,55 (2H, ABq, C02CH2-Ar), 7,70 (2H, "d", J-8,8 Hz, nitrofenyl-Hs), 7,97 (2H, "d", J=7,0 Hz, pyridinio-Hs), 8,25 (2H, "d", J-8,8 Hz, nitrofenyl-Hs) og 8,76 (2H, "d", J=7,l Hz, pyridinio-Hs) ppm: IR (nujol) vmaks: 3250 (OH), 1775 (e-laktam), 1700 (ester) og 2.72 (3H, s, -CH3), 3.1-3.7 (5H, m), 3.7-4.3 (2H, m), 4.71 (2H, t, J-6, 3 Hz, -CH2N<®>), 5.15 (1H, d, J-4.9 Hz, OH), 5.20-5.35-5.40-5.55 (2H, ABq, C02CH2- Ar), 7.70 (2H, "d", J-8.8 Hz, nitrophenyl-Hs), 7.97 (2H, "d", J=7.0 Hz, pyridinium-Hs), 8.25 (2H, "d", J-8.8 Hz, nitrophenyl-Hs) and 8.76 (2H, "d", J=7.1 Hz, pyridinio-Hs) ppm: IR (nujol) vmax: 3250 ( OH), 1775 (e-lactam), 1700 (ester) and
1625 (pyridinio) cm-<1>; 1625 (pyridinium) cm-<1>;
UV (abs, EtOH) Xmaks: 308 (c4,47 x 10) nm; UV (abs, EtOH) Xmax: 308 (c4.47 x 10) nm;
[a]<g3> +24,8° (c 0,5, MeOH); [α]<g3> +24.8° (c 0.5, MeOH);
Analyse for C24H26N3O6S2CI'B^O: Analysis for C24H26N3O6S2CI'B^O:
Beregnet: C 50,56 H 4,95 N 7,37 Calculated: C 50.56 H 4.95 N 7.37
Funnet: C 50,63 H 4,72 N 6,89 Found: C 50.63 H 4.72 N 6.89
D. ( 5R)- 3- r2-( 4- metvltiopvrldinioletvltlo1- f6S)- f( lRl-hvdroksvetvll- 7- okso- l- azablcyklo( 3. 2. 0) hept- 2- en- 2-karboksylat D. ( 5R )- 3- r2-( 4- methylthiopvrldinioletvlt1- f6S )- f( 1R1-hydroxvetvll- 7- oxo- 1- azablcyclo( 3. 2. 0 ) hept- 2- ene- 2-carboxylate
(5R)-p-nitrofenyl-3-[2-(4-metyltiopyridinio)etyltio]-(6S)-[(IR)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylatklorld (380 mg, 0,688 mmol) ble oppløst i 31,5 ml THF og fosfatbuffer, pH 7,4, (31,5 ml, 0,05 M Fisher) og fortynnet med 31,5 ml Et20. Denne oppløsning ble blandet med 10* Pd-C (380 mg, Engelhard) og hydrogenert ved ca. 2,5 kp/cm<2> på en Parr-ryster ved romtemperatur i 1 time. Det vandige sjikt ble filtrert over Celite for å fjerne katalysa-toren og vasket med 2 x 5 ml H2O. Filtratet og vaskevannet ble forenet og vasket med 2 x 30 ml Et20. Det vandige sjikt ble pumpet av for å fjerne alle organiske oppløsningsmidler og renset ved reversfase-søylekromatografi (Cig-jj-Bondapak, 13 g, Waters Associates), idet det ble eluert med H2O. Fraksjoner med en UV-absorpsjon ved 307 nm ble samlet (ca. 1 (5R)-p-nitrophenyl-3-[2-(4-methylthiopyridinio)ethylthio]-(6S)-[(IR)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept-2-ene -2-Carboxylate Chloride (380 mg, 0.688 mmol) was dissolved in 31.5 mL of THF and phosphate buffer, pH 7.4, (31.5 mL, 0.05 M Fisher) and diluted with 31.5 mL of Et 2 O. This solution was mixed with 10* Pd-C (380 mg, Engelhard) and hydrogenated at approx. 2.5 kp/cm<2> on a Parr shaker at room temperature for 1 hour. The aqueous layer was filtered over Celite to remove the catalyst and washed with 2 x 5 mL H 2 O. The filtrate and washing water were combined and washed with 2 x 30 ml Et 2 O. The aqueous layer was pumped off to remove all organic solvents and purified by reverse phase column chromatography (Cig-jj-Bondapak, 13 g, Waters Associates), eluting with H 2 O. Fractions with a UV absorption at 307 nm were collected (approx. 1
liter) og lyofilisert, hvorved man oppnådde 127 mg (0,334 mmol, utbytte 48,5*) av tittelforbindelsen som et gulaktig pulver. liter) and lyophilized, whereby 127 mg (0.334 mmol, yield 48.5*) of the title compound was obtained as a yellowish powder.
<*>H-NMR (D20, CFT-20) S: 1,20 (3H, d, J-6,4 Hz, l'-CH3), <*>H-NMR (D 2 O, CFT-20) S: 1.20 (3H, d, J-6.4 Hz, 1'-CH 3 ),
2,64 (3H, s, -SCH3), 2,81 (2H, m, -SCH2-), 3,19 (1H, dd, J()_1»=6,l Hz, J6_5-2,6 Hz, 6-H), 3,32 (2H, dd, J-ll Hz, J=5,5 Hz, 4-Hs), 3,92 (1H, dt, J=9,2 Hz, J5_6-2,6 Hz, 5-H), 4,1 (1H, m, 1'-H ), 4,61 (2H, t, J-5,9 Hz, -CH2N<®>), 7,70 (2H, "d" , J-7,1 Hz, aromatisk-Hs) og 8,40 (2H, "d", J=7,l Hz, aromatisk-Hs) ppm; 2.64 (3H, s, -SCH3), 2.81 (2H, m, -SCH2-), 3.19 (1H, dd, J()_1»=6.1 Hz, J6_5-2.6 Hz , 6-H), 3.32 (2H, dd, J-ll Hz, J=5.5 Hz, 4-Hs), 3.92 (1H, dt, J=9.2 Hz, J5_6-2, 6 Hz, 5-H), 4.1 (1H, m, 1'-H ), 4.61 (2H, t, J-5.9 Hz, -CH2N<®>), 7.70 (2H, "d" , J-7.1 Hz, aromatic-Hs) and 8.40 (2H, "d", J=7.1 Hz, aromatic-Hs) ppm;
IR (KBr, plate) vmaks: 3400 (OH), 1750 (e-laktam), 1630 IR (KBr, plate) vmax: 3400 (OH), 1750 (e-lactam), 1630
(pyrldinium) og 1590 (karboksylat) cm-<1>; (pyrrlidinium) and 1590 (carboxylate) cm-<1>;
UV (H20) Xmaks: 231 (c9800) og 307 (c25000) nm; UV (H2O) Xmax: 231 (c9800) and 307 (c25000) nm;
[a]g<3> +3,14° (c 0,5, H20). [a]g<3> +3.14° (c 0.5, H 2 O).
Eksempel 8 Example 8
Fremstilling av 3- r2-( 3- metoksv- l- Pvridinlum) etvltiol- 6a- ri ( R)- hvdroksvetvll- 7- okso- l- azablcvklo( 3. 2. 0) hept- 2- en- 2-karboksvlat A. l-( 2- merkaptoetvl )- 3- metoksypyridirlnmmpt. ansulfonat Preparation of 3- r2-(3-Methoxv-1-Pvridinlum)ethvlthiol-6a-ri(R)-hydroxyvetvll-7-oxo-l-azablcvclo(3.2.0)hept-2-ene-2-carboxvlate A .1-(2-mercaptoethyl)-3-methoxypyridinium mpt. ansulfonate
Til 5°C forkjølt 3-metoksypyridin (698 g, 6,4 mmol) ble det dråpevis satt metansulfonsyre (0,216 ml, 3,05 mmol) og etylensulfid (0,19 ml, 3,2 mmol). Blandingen ble derefter oppvarmet til 60°C i 18 timer, avkjølt til 20°C, fortynnet med 10 ml vann og vasket med 3 x 10 ml eter. Den vandige fasen ble under høyvakuum pumpet i 15 minutter og helt på en Cig-reversfase-kolonne. Tittelforbindelsen ble eluert med vann. De passende fraksjoner ble forenet og dampet inn under høyt vakuum, hvorved man oppnådde det ønskede tiol (61,6 mg, utbytte 76,356). To 5°C precooled 3-methoxypyridine (698 g, 6.4 mmol) methanesulfonic acid (0.216 ml, 3.05 mmol) and ethylene sulfide (0.19 ml, 3.2 mmol) were added dropwise. The mixture was then heated to 60°C for 18 hours, cooled to 20°C, diluted with 10 ml of water and washed with 3 x 10 ml of ether. The aqueous phase was pumped under high vacuum for 15 minutes and poured onto a Cig reverse phase column. The title compound was eluted with water. The appropriate fractions were combined and evaporated under high vacuum to give the desired thiol (61.6 mg, yield 76.356).
IR (CH2C12) <v>maks: 2550 (w, SH) og 1620, 1600, 1585 (m, IR (CH2C12) <v>max: 2550 (w, SH) and 1620, 1600, 1585 (m,
aromatisk) cm-<1>. aromatic) cm-<1>.
<i>H-NMR (DMS0-d6) S: 8,90-7,90 (4H, m, aromatisk C-H), 4,72 <i>H-NMR (DMS0-d6) S: 8.90-7.90 (4H, m, aromatic C-H), 4.72
(2H, t, J-6,6 Hz, CH2N<®>), 4,01 (3H, s, 0CH3), 3,5-3,0 (m, skjult CH2S), 2,66 (1H, dd, J-9,5 Hz, J=7,5 Hz, SH) og 2,31 (3H, s, CH3SO3) ppm. (2H, t, J-6.6 Hz, CH2N<®>), 4.01 (3H, s, 0CH3), 3.5-3.0 (m, hidden CH2S), 2.66 (1H, dd , J-9.5 Hz, J=7.5 Hz, SH) and 2.31 (3H, s, CH3SO3) ppm.
B. p- nltrobenzyl- 3- r2-( 3- metoksy- l- pyridiniumklorid) etyltlol-6a- r1,-( R)- hydroksyetvl1- 7- okso- l- azablcyklo( 3. 2. 0 ) hept- 2-en- 2- karboksylat B. p- niltrobenzyl- 3- r2-( 3- methoxy- l- pyridinium chloride) ethylthlol-6a- r1,-( R)- hydroxyethl1- 7- oxo- l- azablcyclo( 3. 2. 0 ) hept- 2- en- 2- carboxylate
En kald (0°C) oppløsning av p-nitrobenzyl-6a-[l'-(R)-hydroksyetyl]-3,7-diokso-l-azabicyklo(3.2.0)heptan-2-karboksylat (1,04 g, 3 mmol) i 12 ml acetonitril ble behandlet dråpevis med diisopropyletylamin (0,63 ml, 3,6 mmol) og difenylklorfosfat (0,75 ml, 36 mmol) og omrørt ved 0°C i 30 minutter. Det resulterende enol-fosfat ble behandlet med l-(2-merkaptoetyl)-3-metoksypyridiniummetansulfonat (1,14 g, 4,30 mmol) i 7 ml CH3CN, diisopropyletylamin (0,63 ml, ' 4,30 mmol), omrørt i 30 minutter og avkjølt ved -10°C i 30 minutter. Det faste stoff som ble felt ut fra blandingen, ble filtrert av, vasket med 2 ml kald acetonitril og tørket under dannelse av tittelforbindelsen (1,32 g, utbytte 82*). A cold (0°C) solution of p-nitrobenzyl-6α-[1'-(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-carboxylate (1.04 g , 3 mmol) in 12 mL of acetonitrile was treated dropwise with diisopropylethylamine (0.63 mL, 3.6 mmol) and diphenylchlorophosphate (0.75 mL, 36 mmol) and stirred at 0 °C for 30 min. The resulting enol phosphate was treated with 1-(2-mercaptoethyl)-3-methoxypyridinium methanesulfonate (1.14 g, 4.30 mmol) in 7 mL of CH 3 CN, diisopropylethylamine (0.63 mL, 4.30 mmol), stirred for 30 minutes and cooled at -10°C for 30 minutes. The solid which precipitated from the mixture was filtered off, washed with 2 ml of cold acetonitrile and dried to give the title compound (1.32 g, yield 82*).
IR (nujol) vmaks: 3320 (m, OH), 1780, 1765 (s, e-laktam C=0), IR (nujol) vmax: 3320 (m, OH), 1780, 1765 (s, ε-lactam C=0),
1700, 1695 (m, ester C=0) og 1520 (s, N02) cm"<1>; 1700, 1695 (m, ester C=0) and 1520 (s, NO 2 ) cm"<1>;
<1>H-NMR (DMS0-d6) S: 9,01 (1H, bs, H-3 aromatisk), 8,75 (1H, <1>H-NMR (DMS0-d6) S: 9.01 (1H, bs, H-3 aromatic), 8.75 (1H,
bd, J-5,4 Hz, H-6 aromatisk), 8,35-7,95 (4H, m, H-aromatisk), 7,70 (2H, d, J-7,7 Hz, H-aromatisk), 5,37 (2H, sentrum av ABq, J-13 Hz, CH2pNB), 5,17 (1H, d, J-4,9 Hz, OH), 4,87 (2H, t, J-6,3 Hz, CH2-N<®>), 4,35-3,75 (2H, m, H-5 ogH-1'), 4,00 (3H, s, OCH3), 3,56 (del av en t, J-6,3 Hz, CH2S), 3,5-3,20 (3H, m, H-6, H-3) og 1,16 (3H, d, J-6,1 Hz, CH3CHO) ppm. bd, J-5.4 Hz, H-6 aromatic), 8.35-7.95 (4H, m, H-aromatic), 7.70 (2H, d, J-7.7 Hz, H-aromatic ), 5.37 (2H, center of ABq, J-13 Hz, CH2pNB), 5.17 (1H, d, J-4.9 Hz, OH), 4.87 (2H, t, J-6, 3 Hz, CH2-N<®>), 4.35-3.75 (2H, m, H-5 andH-1'), 4.00 (3H, s, OCH3), 3.56 (part of a t, J-6.3 Hz, CH2S), 3.5-3.20 (3H, m, H-6, H-3) and 1.16 (3H, d, J-6.1 Hz, CH3CHO) ppm.
C. 3- r2-( 3- metoksy- l- pyridinlum) etyltlo1- 6a- fl'-( R)- hydroksy-etvll- 7- okso- l- azabicyklo( 3. 2. 0) hept- 2- en- 2- karboksylat C. 3- r2-( 3- methoxy-l- pyridinelum) ethyltlo1- 6a- fl'-( R)- hydroxy-etvll- 7- oxo- l- azabicyclo( 3. 2. 0) hept- 2- en- 2- carboxylate
En oppløsning av p-nitrobenzyl-3-[2-(3-metoksy-l-pyridiniumklorid )etyltio]-6a-[1'-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat (600 mg, 1,12 mmol) i 25 ml THF, 25 ml eter og fosfatbuffer, pH 7,4 (0,1 M, 25 ml) ble hydrogenert i en Parr-ryster over 10* Pd/C (1,1 g) i 1 time ved ca. 2,8 kp/cm2 . Blandingen ble fortynnet med eter og den vandige fase filtrert gjennom herdet filterpapir nr. 52. Det vandige sjikt ble vasket med 2 x 20 ml eter, pumpet under vakuum og helt på en silikagel-reversfase-kolonne. Tittelforbindelsen ble eluert med vann inneholdende 2 og 5* acetonitril. De passende fraksjoner ble forenet og lyofilisert under dannelse av et gult faststoff, som igjen ble renset ved hjelp av HPLC under dannelse av penem-karboksy-latet (150 mg, 38*); A solution of p-nitrobenzyl-3-[2-(3-methoxy-1-pyridinium chloride)ethylthio]-6a-[1'-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept -2-ene-2-carboxylate (600 mg, 1.12 mmol) in 25 mL THF, 25 mL ether and phosphate buffer, pH 7.4 (0.1 M, 25 mL) was hydrogenated in a Parr shaker over 10 * Pd/C (1.1 g) for 1 hour at approx. 2.8 kp/cm2. The mixture was diluted with ether and the aqueous phase was filtered through hardened filter paper No. 52. The aqueous layer was washed with 2 x 20 ml of ether, pumped under vacuum and poured onto a silica gel reverse phase column. The title compound was eluted with water containing 2 and 5* acetonitrile. The appropriate fractions were combined and lyophilized to give a yellow solid, which was again purified by HPLC to give the penem carboxylate (150 mg, 38*);
IR (nujol) vmaks: 1750 (s, e-laktam C=0) og 1580 (s, IR (nujol) vmax: 1750 (s, e-lactam C=0) and 1580 (s,
karboksylat) cm-<1>; carboxylate) cm-<1>;
<1->H-NMR (D20) S: 8,55-8,10 (2H, m, H-2, H-6 aromatisk), 8,17-7,75 (2H, m, H-3, H-4 aromatisk), 4,77 (2H, t, J=5,9 Hz, CH2N<e>), 4,10 (1H, del av 5 linjer, J=»6,3 Hz, <1->H-NMR (D2O) S: 8.55-8.10 (2H, m, H-2, H-6 aromatic), 8.17-7.75 (2H, m, H-3, H-4 aromatic), 4.77 (2H, t, J=5.9 Hz, CH2N<e>), 4.10 (1H, part of 5 lines, J=»6.3 Hz,
H-l'), 3,97 (3H, s, OCH3), 3,85, 3,82 (2 linjer, del av dt, J-2,6 Hz, del av H-5), 3,42 (2H, t, J-5,9 Hz, CH2-S), 3,25 (1H, dd, J-6,1 Hz, J-2,6 Hz, H-6), 2,99-2,60 (2H, 6 linjer, del av H-3) og 1,20 (3H, H-l'), 3.97 (3H, s, OCH3), 3.85, 3.82 (2 lines, part of dt, J-2.6 Hz, part of H-5), 3.42 ( 2H, t, J-5.9 Hz, CH2-S), 3.25 (1H, dd, J-6.1 Hz, J-2.6 Hz, H-6), 2.99-2.60 (2H, 6 lines, part of H-3) and 1.20 (3H,
d, J-6,4 Hz, CH3) ppm; d, J-6.4 Hz, CH 3 ) ppm;
UV (H20, c 0,05) Xmaks: 290 (cl0517), 223 (c6643) nm; UV (H 2 O, c 0.05) Xmax: 290 (cl0517), 223 (c6643) nm;
t% (0,1 M fosfatbuffer, pH 7,4, 37<*>C): 20 t. t% (0.1 M phosphate buffer, pH 7.4, 37<*>C): 20 h.
Eksempel 9 Example 9
Fremstilling av ( 5R. 6S)- 3- ff2-( 3- metvltiopvridlniobetviltiol-6- fl-( R )- hvdroksvetvl" l- 7- okso- l- azabicvklo( 3. 2. 0 ) hept- 2- en- 2-karboksylat Preparation of (5R.6S)-3-ff2-(3-methylthiopvridlniobetylthiol-6-fl-(R)-hydroxymethyl"l-7-oxo-l-azabiclo(3.2.0)hept-2-ene-2 -carboxylate
A. 3- metvltio- l-( 2- merkaptoetyl) pyrldiniumklorid Til en oppløsning av 3-metyltiopyridin<3> (2,00 g, 0,016 mol) i 10 ml eter settes 15 ml IN HC1 og det hele rystes grundig. Den vandige fase separeres av, vaskes med 10 ml eter og dampes inn. Det gjenblevne hydrokloridet tørkes derefter i vakuum over P205 under dannelse av et hvitt, fast stoff. Til dette faste hydrokloridet settes 3-metyltiopyridin (1,88 g, <3> Fremstilt ved metoden ifølge J.A. Zoltewiez og C. A. 3-Methylthio-1-(2-mercaptoethyl) pyrldinium chloride To a solution of 3-methylthiopyridine<3> (2.00 g, 0.016 mol) in 10 ml of ether is added 15 ml of 1N HCl and the whole is shaken thoroughly. The aqueous phase is separated off, washed with 10 ml of ether and evaporated. The remaining hydrochloride is then dried in vacuo over P 2 O 5 to give a white solid. To this solid hydrochloride is added 3-methylthiopyridine (1.88 g, <3> Prepared by the method according to J.A. Zoltewiez and C.
Nisi, "J. Org. Chem.", 34, 765 (1969). Nisi, "J. Org. Chem.", 34, 765 (1969).
0,015 mol) og etylensulfid (0,89 ml, 0,015 mol), og den resulterende blanding ble oppvarmet i et oljebad til 55-65°C under ?2 i 15 timer. Dette ga en lett uklar olje som ble samlet i 125 ml H20 og vasket med CH2C12. Den vandige oppløsning ble konsentrert til ca. 2 ml og derefter ble det tilsatt noen få dråper acetonitril for å gjøre blandingen homogen. Den resulterende vandige oppløsning ble satt til en C^g-reversfase-kolonne. Eluering med H20 og efterfølgende inndamping av de relevante fraksjoner ga produktet (2,66 g, 80*) som en blekgul, viskøs olje. 0.015 mol) and ethylene sulfide (0.89 mL, 0.015 mol), and the resulting mixture was heated in an oil bath to 55-65°C under ?2 for 15 hours. This gave a slightly cloudy oil which was taken up in 125 ml H 2 O and washed with CH 2 Cl 2 . The aqueous solution was concentrated to approx. 2 ml and then a few drops of acetonitrile were added to make the mixture homogeneous. The resulting aqueous solution was added to a C₂g reverse phase column. Elution with H 2 O and subsequent evaporation of the relevant fractions gave the product (2.66 g, 80*) as a pale yellow viscous oil.
IR (film) vmaks: 2410 (br, -SH) cm-<1>;IR (film) vmax: 2410 (br, -SH) cm-<1>;
^H-NMR (DMS0-d6 + D20) S: 8,88-7,88 (m, 4H, aromatisk), 4,70 ^H-NMR (DMS0-d6 + D2O) S: 8.88-7.88 (m, 4H, aromatic), 4.70
(t, J=6,5 Hz, 2H, N-CH2), 3,08 (skrå t, J-6,5 Hz, • 2H, S-CH2), 2,64 (s, 3H, S-Me). (t, J=6.5 Hz, 2H, N-CH2), 3.08 (oblique t, J-6.5 Hz, • 2H, S-CH2), 2.64 (s, 3H, S-Me ).
B. p- nitrobenzvl-( 5R. 6S)- 3- r2-( 3- metvltiopvridinio) etvltio" l-6- f1-( R)- hvdroksvetvn- 7- okso- l- azabicvklo( 3. 2. 0 ) hept- 2-en- 2- karboksylatklorid B. p- nitrobenzvl-( 5R. 6S)- 3- r2-( 3- methylthiopvridinio) etvlthio" 1-6- f1-( R)- hydroxymethyl- 7- oxo- 1- azabicclo( 3. 2. 0 ) hept - 2-ene-2- carboxylate chloride
En oppløsning av p-nitrobenzyl-(5R,6S)-6-[l-(R)-hydroksy-etyl]-3,7-diokso-l-azabicyklo(3.2.0)heptan-2-karboksylat (0,522 g, 1,50 mmol) i 7 ml tørr acetonitril avkjøles til 0°C og derefter tilsettes dråpevis diisopropyletylamin (0,287 ml, 1,65 mmol). Til den resulterende gulbrune oppløsning ble det dråpevis satt difenylklorfosfat (0,342 ml, 1,65 mmol) og reaksjonsblandingen ble holdt ved 0°C i 30 minutter. Derefter ble det tilsatt diisopropyletylamin (0,313 ml, 1,80 mmol), fulgt av en oppløsning av 3-metyltlo-l-(2-merkaptoetyl)-pyridiniumklorid (0,398 g, 1,80 mmol) i 0,70 ml tørr DMF. Ca. A solution of p-nitrobenzyl-(5R,6S)-6-[1-(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-carboxylate (0.522 g, 1.50 mmol) in 7 ml of dry acetonitrile is cooled to 0°C and then diisopropylethylamine (0.287 ml, 1.65 mmol) is added dropwise. To the resulting tan solution was added dropwise diphenylchlorophosphate (0.342 mL, 1.65 mmol) and the reaction mixture was kept at 0°C for 30 minutes. Diisopropylethylamine (0.313 mL, 1.80 mmol) was then added, followed by a solution of 3-methylthio-1-(2-mercaptoethyl)pyridinium chloride (0.398 g, 1.80 mmol) in 0.70 mL of dry DMF. About.
1 minutt efter avsluttet tilsetning skilte det seg ut et bunnfall fra reaksjonsblandingen og ytterligere avkjøling ved 1 minute after the end of the addition, a precipitate separated from the reaction mixture and further cooling by
-10°C i 10 minutter ga en fast, oransjefarvet masse. Dette faste stoff ble derefter triturert med acetonitril og resten samlet ved filtrering. Resten ble vasket med acetonitril, derefter med aceton og til slutt tørket i vakuum og man oppnådde 0,455 g tilsvarende 55* av produktet som et kremfarvet, fast stoff. Det forenede filtrat ble dampet inn under dannelse av en gul olje som ble samlet i et minimalt volum acetonitril og avkjølt ved 0°C i 30 minutter. Filtrering av denne blanding ga ytterligere 0,139 g produkt som et lysegult, fast stoff. Det samlede utbytte var 0,594 g (72*). IR (KBr) vmaks: 3345 (br, -GH), 1770 (e-laktam CO), 1680 (-CO2PNB) cm-1; ^H-NMR (DMSO-dfc) S: 8,98-7,96 (m, 4H, pyrldinium aromatisk), 8,20-7,65 (ABq, J-7,0 Hz, 4H, pNB aromatisk), 5,53-4,80 (m, 4H), 4,3-3,7 (m, 2H), 3,6-3,25 (m, 6H), 2,66 (s, 3H, S-Me), 1,16 (d, J-6,0 Hz, 3H, CHMe). C. ( 5R. 6S)- 3- r2-( 3- metvltiopvrldinio) etvltlo1- 6- ri-( R)-hvdroksyetvll- 7- okso- l- azabicvklo( 3. 2. 0) hept- 2- en- 2-karboks<y>lat -10°C for 10 minutes gave a firm, orange colored mass. This solid was then triturated with acetonitrile and the residue collected by filtration. The residue was washed with acetonitrile, then with acetone and finally dried in vacuo and 0.455 g corresponding to 55% of the product was obtained as a cream-colored solid. The combined filtrate was evaporated to give a yellow oil which was collected in a minimal volume of acetonitrile and cooled at 0°C for 30 minutes. Filtration of this mixture gave an additional 0.139 g of product as a pale yellow solid. The overall yield was 0.594 g (72*). IR (KBr) vmax: 3345 (br, -GH), 1770 (e-lactam CO), 1680 (-CO2PNB) cm-1; ^H-NMR (DMSO-dfc) S: 8.98-7.96 (m, 4H, pyrldinium aromatic), 8.20-7.65 (ABq, J-7.0 Hz, 4H, pNB aromatic), 5.53-4.80 (m, 4H), 4.3-3.7 (m, 2H), 3.6-3.25 (m, 6H), 2.66 (s, 3H, S-Me ), 1.16 (d, J-6.0 Hz, 3H, CHMe). C. ( 5R. 6S)- 3- r2-( 3- metvlthiopvrldinio) etvltlo1- 6- ri-( R)-hydroxyetvll- 7- oxo- 1- azabicclo( 3. 2. 0) hept- 2- en- 2 -karboks<y>lat
Til en blanding av p-nitrobenzyl-(5R,6S)-3-[2-(3-metyltio-pyridinio)etyltio]-6-[l-(R)-hydroksyetyl]-7-okso-1-azabicyklo(3.2.0)hept-2-en-2-karboksylatklorid (0,551 g, 1,0 mmol) og 0,55 g 10* palladium-på-trekull i 25 ml fosfatbuffer (0,05 M, pH 7,4) ble det satt 5 ml THF og 25 ml eter. Denne blanding ble hydrogenert (Parr) ved ca. 2,8 kp/cm<2> i 1 time. Reaksjonsblandingen ble derefter filtrert gjennom Celite og filterkaken vaskes med H2O og eter. Den vandige fase ble separert og vasket ytterligere tre ganger med eter. Efter fjerning av gjenværende organiske oppløsningsmidler i vakuum, ble den vandige oppløsning avkjølt til 0<*>C og pH-verdien innstilt til 7,0 med mettet vandig NaHCC^. Denne oppløsning ble øyeblikkelig satt til en Cig-reversfase-kolonne. Eluering med H2O og efterfølgende lyofilisering av de relevante fraksjoner ga 0,25 g av et klart gult, fast stoff. Dette materialet ble renset igjen ved reversfase-HPLC og man oppnådde 0,210 g tilsvarende 55* av produktet som et lysegult, fast stoff. To a mixture of p-nitrobenzyl-(5R,6S)-3-[2-(3-methylthio-pyridinio)ethylthio]-6-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2 .0)hept-2-ene-2-carboxylate chloride (0.551 g, 1.0 mmol) and 0.55 g of 10* palladium-on-charcoal in 25 ml of phosphate buffer (0.05 M, pH 7.4) gave added 5 ml of THF and 25 ml of ether. This mixture was hydrogenated (Parr) at approx. 2.8 kp/cm<2> for 1 hour. The reaction mixture was then filtered through Celite and the filter cake washed with H2O and ether. The aqueous phase was separated and washed three more times with ether. After removal of residual organic solvents in vacuo, the aqueous solution was cooled to 0<*>C and the pH adjusted to 7.0 with saturated aqueous NaHCO3. This solution was immediately applied to a Cig reverse phase column. Elution with H 2 O and subsequent lyophilization of the relevant fractions gave 0.25 g of a clear yellow solid. This material was purified again by reverse phase HPLC and 0.210 g corresponding to 55* of the product was obtained as a pale yellow solid.
IR (KBr) vmaks: 3400 (br, -OH), 1755 (e-laktam CO), 1590 IR (KBr) vmax: 3400 (br, -OH), 1755 (e-lactam CO), 1590
(-C02<e>) cm"<1>; (-C02<e>) cm"<1>;
iB-NMR (D20) S: 8,60-7,76 (m, 4H, aromatisk), 4,76 (t, iB-NMR (D20) S: 8.60-7.76 (m, 4H, aromatic), 4.76 (t,
J-5,8 Hz, 2H, N-CH2), 4,13 (d av q, «W-6,3 Hz, 1H, H-l')t 3,95 (d av t, J-9,0 Hz, J'-2,8 Hz, 1H, H-5), 3,45-2,75 (m, 5H), 2,59 (s, 3H, S-Me), 1,20 (d, J=6,4 Hz, 3H, CHMe); J-5.8 Hz, 2H, N-CH2), 4.13 (d of q, «W-6.3 Hz, 1H, H-l')t 3.95 (d of t, J-9, 0 Hz, J'-2.8 Hz, 1H, H-5), 3.45-2.75 (m, 5H), 2.59 (s, 3H, S-Me), 1.20 (d, J=6.4 Hz, 3H, CHMe);
UV (H20) <X>maks: 296 (c8509), 273 (cl3005), 231 (cll576) nm; UV (H 2 O) <X>max: 296 (c8509), 273 (cl3005), 231 (cll576) nm;
ttø (pH 7,4, 36,8°C): 20 t. tthaw (pH 7.4, 36.8°C): 20 h.
Eksempel 10 Example 10
Fremstilling av 3- f2-( l-( 2. 6- dimetvlPvrldinium) etvltlo1- 6a-ri-( R)- hvdroksvetvl1- 7- okso- l- azablcyklo( 3. 2. 0) hept- 2- en- 2-karboksvlat Preparation of 3- f2-( l-( 2. 6- dimethylPvrldinium) etvltlo1- 6a-ri-( R)- hvdroxvetl1- 7- oxo- l- azablcyclo( 3. 2. 0) hept- 2- en- 2- carboxylate
A. l-( 2- merkaptoetvl)- 2. 6- dlmetvlpvridiniummetansulfonat A. 1-(2-mercaptoethyl)-2.6-dlmethylpyridinium methanesulfonate
En blanding av 2,6-dimetylpyridin (19,2 ml, 0,165 mol) og metansulfonsyre (3,27 ml, 0,050 mol) ble omrørt i 15 minutter, behandlet med etylensulfid (4,17 ml, 0,070 mol) og omrørt ved 100°C i 42 timer under en nitrogenatmosfaere. Efter avkjøling til 25°C ble reaksjonsblandingen fortynnet med 45 ml eter og 30 ml vann. De to sjikt ble separert og det organiske sjikt ekstrahert med 2 x 5 ml vann. De vandige sjikt ble forenet, filtrert gjennom en Celite-pute, vasket med 2 x 15 ml eter, pumpet for å fjerne spor av organiske oppløsningsmidler og helt på toppen av en 3,0 x 12 cm jj-Bondapak C^g-kolonne. Eluerlng med en blanding av 3* acetonitril og 97* vann ga efter lyofilisering av de passende fraksjoner 2,5 g av den urene tittelforbindelse som en sirup. Den renses igjen ved hjelp av HPLC (ji-Bondapak C^g), hvorved man oppnådde 0,90 g tilsvarende 7* av tittelforbindelsen. A mixture of 2,6-dimethylpyridine (19.2 mL, 0.165 mol) and methanesulfonic acid (3.27 mL, 0.050 mol) was stirred for 15 min, treated with ethylene sulfide (4.17 mL, 0.070 mol) and stirred at 100 °C for 42 hours under a nitrogen atmosphere. After cooling to 25°C, the reaction mixture was diluted with 45 ml of ether and 30 ml of water. The two layers were separated and the organic layer extracted with 2 x 5 ml of water. The aqueous layers were combined, filtered through a pad of Celite, washed with 2 x 15 mL of ether, pumped to remove traces of organic solvents, and poured onto a 3.0 x 12 cm jj-Bondapak C^g column. Elution with a mixture of 3% acetonitrile and 97% water after lyophilization of the appropriate fractions gave 2.5 g of the impure title compound as a syrup. It is purified again by means of HPLC (ji-Bondapak C^g), whereby 0.90 g corresponding to 7* of the title compound was obtained.
IR (film) vmaks: 2520 (SH), 1640 og 1625 (pyrldinium), 1585, IR (film) vmax: 2520 (SH), 1640 and 1625 (pyrldinium), 1585,
1490, 1200 (sulfonat) cm-<1>; 1490, 1200 (sulfonate) cm-<1>;
<1->H-NMR (DMS0-d6 + D20) S: 2,36 (3H, s, CH2S03<e>), 4,62 (2H, <1->H-NMR (DMS0-d6 + D2O) S: 2.36 (3H, s, CH2SO3<e>), 4.62 (2H,
m, CH2N<®>), 7,74 (2H, m, Hm av pyrldinium), 8,24 (1H, m, Hp av pyrldinium); m, CH 2 N<®>), 7.74 (2H, m, Hm of pyrldinium), 8.24 (1H, m, Hp of pyrldinium);
UV (H20) Xmaks: 272 (4080) nm. UV (H20) Xmax: 272 (4080) nm.
B. p- nitrobenzvl- 3- r2-( l-( 2. 6- dimetylpyrldlnlum)) etyltlo1- 6a-l" l-( R )- hydroksyetyl" l- 7- okso- l- azabicyklo( 3. 2. 0) hept- 2- en-karboksylat- dlfenvlfosfat B. p-nitrobenzvl-3-r2-(l-(2.6-dimethylpyrldlnlum))ethyltlo1-6a-l"l-(R)- hydroxyethyl"l-7- oxo-l- azabicyclo(3.2.0 ) heptane-2-ene-carboxylate-diphenylphosphate
Til en kald (0'C) oppløsning av p-nitrobenzyl-6a-[l-(R)-hydroksyetyl]-3,7-diokso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat (0,658 g, 1,89 mmol) i 6 ml acetonitril, som holdes under en nitrogenatmosfære, ble det satt diisopropyletylamin (0,394 ml, 2,26 mmol) og difenylklorfosfat (0,468 ml, 2,26 mmol). Reaksjonsblandingen ble omrørt i 30 minutter og derefter behandlet med en oppløsning av l-(2-merkaptoetyl)-2,6-dimetylpyridinlummetansulfonat (0,720 g, 2,73 mmol) i 3 ml acetonitril, fulgt av diisopropyletylamin (0,394 ml, 2,26 mmol). Reaksjonsblandingen ble omrørt ved 0<*>C i 2 timer, fortynnet med 27 ml kaldt vann av 0<*>C og helt på toppen av en 25 x 9,0 cm kolonne av ji-Bondapak C^g. Eluerlng med blan-dinger av acetonitril og vann og lyofilisering av de passende fraksjoner ga 0,92 g tilsvarende 655É av tittelforbindelsen. To a cold (0'C) solution of p-nitrobenzyl-6α-[l-(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)hept-2-ene-2-carboxylate ( 0.658 g, 1.89 mmol) in 6 mL of acetonitrile, which is kept under a nitrogen atmosphere, was added diisopropylethylamine (0.394 mL, 2.26 mmol) and diphenylchlorophosphate (0.468 mL, 2.26 mmol). The reaction mixture was stirred for 30 minutes and then treated with a solution of 1-(2-mercaptoethyl)-2,6-dimethylpyridinelumethanesulfonate (0.720 g, 2.73 mmol) in 3 mL of acetonitrile, followed by diisopropylethylamine (0.394 mL, 2.26 mmol). The reaction mixture was stirred at 0<*>C for 2 hours, diluted with 27 mL of cold 0<*>C water and poured onto the top of a 25 x 9.0 cm column of ji-Bondapak C^g. Elution with mixtures of acetonitrile and water and lyophilization of the appropriate fractions yielded 0.92 g corresponding to 655E of the title compound.
IR (KBr) vmaks: 3700-3000 (OH), 1765 (C-0 av e-laktam), 1690 IR (KBr) vmax: 3700-3000 (OH), 1765 (C-0 of ε-lactam), 1690
(C-0 av pNB-ester), 1620 (pyrldinium), 1590 (fenyl), 1517 (N02), 1330 (N02), 880 (N02) cm"<1>; (C-0 of pNB ester), 1620 (pyrldinium), 1590 (phenyl), 1517 (NO 2 ), 1330 (NO 2 ), 880 (NO 2 ) cm"<1>;
<1->H-NMR (DMS0-d6) S: 1,15 (3H, d, J-6,2 Hz, CH3CH0H), 2,7-3,7 (11H, CH2S, 2-CH3 på pyrldinium, H-4, H-6), 3,7-4,4 (2H, CH3CHOH, H-5), 4,7 (2H, m, CH2N<®>), 5,14 (1H, d, J-4,5 Hz, OH), 5,37 (sentrum av ABq, J-13,2 Hz, CH2 av pNB), 6,7-7,5 (10H, m, fenyl), 7,5-8,7 (7H, pyrldinium, H'er av pNB); <1->H-NMR (DMS0-d6) S: 1.15 (3H, d, J-6.2 Hz, CH3CH0H), 2.7-3.7 (11H, CH2S, 2-CH3 on pyrldinium, H-4, H-6), 3.7-4.4 (2H, CH3CHOH, H-5), 4.7 (2H, m, CH2N<®>), 5.14 (1H, d, J- 4.5 Hz, OH), 5.37 (center of ABq, J-13.2 Hz, CH2 of pNB), 6.7-7.5 (10H, m, phenyl), 7.5-8.7 (7H, pyrldinium, H's of pNB);
UV (H20) <X>maks: 274 (cl4150), 319 (c9445) nm. UV (H20) <X>max: 274 (cl4150), 319 (c9445) nm.
C. 3- T2-( l-( 2. 6- dlmetvlpvrldinium)) etvltio1- 6a- fl-( R)-hvdroksvetvll- 7- okso- l- azablcvklo( 3. 2. 0) hept- 2- en- 2-karboksvlat C. 3- T2-( l-( 2. 6- dlmetvlpvrldinium)) etvltio1- 6a- fl-( R)-hydroxylvetvll- 7- oxo- l- azablcvclo( 3. 2. 0) hept- 2- en- 2 - carbox vat
Til en blanding av p-nitrobenzyl-3-[2-(l-(2,6-dimetylpyri-dinium))etyltio]-6a-[1-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat-difenylfosfat (0,80 g, 1,07 mmol) i 42 ml våt tetrahydrofuran ble det satt 42 ml eter, monobasisk kaliumfosfat-natriumhydroksydbuffer (0,15 M, pH 7,22, 21 ml) og 0,80 g 10* palladium-på-trekull. Den resulterende blanding ble hydrogenert i 1 time under ca. 2,8 kp/cm<2> ved 23°C og filtrert gjennom en Celite-pute. De to sjikt ble separert og det organiske sjikt ekstrahert med 3 x 8 ml buffer. Den vandige fase ble forenet, vasket med 50 ml eter, pumpet for å fjerne spor av organisk oppløsnings-middel og helt på toppen av en 3,0 x 10,2 cm p-Bondapak C^g-kolonne. Eluering av kolonnen med en blanding av 5* acetonitril og 95* vann og lyofilisering av de passende fraksjoner ga tittelforbindelsen i en mengde av 0,246 g tilsvarende 63* som et gulaktig pulver. To a mixture of p-nitrobenzyl-3-[2-(1-(2,6-dimethylpyridinium))ethylthio]-6a-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2 .0)hept-2-ene-2-carboxylate diphenyl phosphate (0.80 g, 1.07 mmol) in 42 ml of wet tetrahydrofuran was added 42 ml of ether, monobasic potassium phosphate-sodium hydroxide buffer (0.15 M, pH 7, 22, 21 ml) and 0.80 g of 10* palladium-on-charcoal. The resulting mixture was hydrogenated for 1 hour under approx. 2.8 kp/cm<2> at 23°C and filtered through a Celite pad. The two layers were separated and the organic layer extracted with 3 x 8 ml of buffer. The aqueous phase was combined, washed with 50 ml of ether, pumped to remove traces of organic solvent and poured onto a 3.0 x 10.2 cm p-Bondapak C 2 g column. Elution of the column with a mixture of 5* acetonitrile and 95* water and lyophilization of the appropriate fractions gave the title compound in an amount of 0.246 g corresponding to 63* as a yellowish powder.
IR (KBr) vmaks: 3700-2800 (0H), 1750 (C-0 av e-laktam), 1620 IR (KBr) vmax: 3700-2800 (0H), 1750 (C-0 of e-lactam), 1620
(pyrldinium), 1585 (karboksylat) cm-<1>; (pyrrlidinium), 1585 (carboxylate) cm-<1>;
^H-NMR (D20) S: 1,23 (3H, d, J=6,4 Hz, CH3CH0H), 2,5-3,5 ^H-NMR (D 2 O) S: 1.23 (3H, d, J=6.4 Hz, CH 3 CH 0 H), 2.5-3.5
(11H, H-4, H-6, CH2S, 2CH3 på pyrldinium), 3,8-4,4 (2H, CH3CHOH, H-5), 4,5-4,9 (CH2N<®>, HOD), 7,64 og 7,74 (2H, A del av A2B-system, Hm av pyrldinium), 8,07, 8,16, 8,18 og 8,27 (1H, B del av A2B-system, Hp av pyrldinium); (11H, H-4, H-6, CH2S, 2CH3 on pyrldinium), 3.8-4.4 (2H, CH3CHOH, H-5), 4.5-4.9 (CH2N<®>, HOD) , 7.64 and 7.74 (2H, A part of A2B system, Hm of pyrldinium), 8.07, 8.16, 8.18 and 8.27 (1H, B part of A2B system, Hp of pyrldinium);
UV (H20) <X>maks: 277 (c9733), 300 (c8271) nm; UV (H 2 O) <X>max: 277 (c9733), 300 (c8271) nm;
[a]<g3> +50,7° (c 0,48, H20); [α]<g3> +50.7° (c 0.48, H 2 O);
Analyse for Ci<g>H22<N>204S*1,5H20: Analysis for Ci<g>H22<N>204S*1.5H20:
Eksempel 11 Example 11
Fremstilling av ( 5R. 6S)- 3- T2-( 2- metyltlo- 3- metyllmidazolio1-etvltio1- 6- ri-( R)- hvdroksvetvn- 7- okso- l- azabicvklo( 3. 2. 01-hept- 2- en- 2- karboksvlat Preparation of (5R.6S)-3-T2-(2-methyltlo-3-methylmidazolio1-ethylthio1-6-ri-(R)-hydroxymethyl-7-oxo-1-azabicyclo(3.2.01-hept-2 - a- 2- carboxylate
A. 2- metyltio- 3- metyl- l-( 2- merkaptoetylUmldazoliumtrifluor-metansulfonat A. 2- Methylthio- 3- methyl- 1-( 2- mercaptoethyl Umldazolium trifluoromethanesulfonate
Trifluormetansulfonsyre (1,38 ml, 0,015 mol) ble dråpevis satt til 2-metyltio-l-metylimidazol<4> (4,0 g, 0,03 mol) ved 0°C under N2. Derefter ble det tilsatt etylensulfid (0,9 ml, 0,015 mol) og blandingen ble oppvarmet til 55°C under N2 i 24 timer. Reaksjonsblandingen ble triturert tre ganger med eter og resten samlet i aceton, filtrert og dampet inn. Dette ga produktet i en mengde av 4,2 g tilsvarende 82* som et semikrystallinsk, fast stoff som ble anvendt som sådant uten ytterligere rensing. Trifluoromethanesulfonic acid (1.38 mL, 0.015 mol) was added dropwise to 2-methylthio-1-methylimidazole<4> (4.0 g, 0.03 mol) at 0°C under N 2 . Ethylene sulfide (0.9 mL, 0.015 mol) was then added and the mixture was heated to 55°C under N 2 for 24 h. The reaction mixture was triturated three times with ether and the residue collected in acetone, filtered and evaporated. This gave the product in an amount of 4.2 g corresponding to 82* as a semi-crystalline solid which was used as such without further purification.
IR (film) vmaks: 2550 (w, sh) cm"1; <4> Fremstilt som i A. Wohl og W. Marckwald, "Chem. IR (film) vmax: 2550 (w, sh) cm"1; <4> Prepared as in A. Wohl and W. Marckwald, "Chem.
Ber.", 22, 1353 (1889). Ber.", 22, 1353 (1889).
<1>H-NMR (dfc-acetcm) S: 7,97 (s, 2H), 4,66 (t, J-7 Hz, 2H, <1>H-NMR (dfc-acetcm) S: 7.97 (s, 2H), 4.66 (t, J-7 Hz, 2H,
metylen), 4,17 (s, 3H, N-Me), 3,20 (d av t, J-7 Hz, J'-9 Hz, 2H, metylen), 2,72 (s, 3H, S-Me), 2,20 (t, J-9 Hz, 1H, -SH). methylene), 4.17 (s, 3H, N-Me), 3.20 (d of t, J-7 Hz, J'-9 Hz, 2H, methylene), 2.72 (s, 3H, S- Me), 2.20 (t, J-9 Hz, 1H, -SH).
B. p- nltrobenzvl-( 5R. 6S)- 3- r2-( 2- metyltlo- 3- metyl- imlda-zolio letvltlol - 6- fl- f R l- hvdroksvetvll - 7- okso- l- azablcyklo-( 3. 2. 0) hept- 2- en- 2- karboksvlat- dlfenvlfosfat B. p- nltrobenzvl-( 5R. 6S)- 3- r2-( 2- methyltlo- 3- methyl- imlda-zolio letvltlol - fl- f R l- hvdroxvetvll - 7- oxo- l- azablcyclo-( 3 .
Til en oppløsning av p-nitrobenzyl-(5R,6S)-6-[l-(R)-hydroksy-etyl]-3,7-diokso-l-azabicyklo(3.2.0)heptan-2-karboksylat (1,40 g, 4,0 mmol) i 50 ml tørr acetonitril ved 0°C under N2 ble det dråpevis satt diisopropyletylamin (0,76 ml, 4,4 mmol) fulgt av difenylklorfosfat (0,91 ml, 4,1 mmol). Efter omrøring av reaksjonsblandingen ved romtemperatur i 1 time ble det tilsatt diisopropyletylamin (0,76 ml, 4,4 mmol) og derefter dråpevis en oppløsning av 2-metyltio-3-metyl-l-(2-merkaptoetyl )imidazolium-trifluormetansulfonat (2,0 g, 5,9 mmol) i 5 ml acetonitril. Reaksjonsblandingen ble holdt ved romtemperatur i VA time og derefter konsentrert i vakuum for å oppnå en gummi. Denne ble samlet i H20, derefter med 20* acetonitrll-H20 og til slutt 30* acetonitril-H20, fulgt av lyofilisering av de passende fraksjoner ga produktet i en mengde av 0,90 g tilsvarende 30* som et lysegult, fast stoff. To a solution of p-nitrobenzyl-(5R,6S)-6-[l-(R)-hydroxy-ethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-carboxylate (1, 40 g, 4.0 mmol) in 50 ml of dry acetonitrile at 0°C under N2 was added dropwise diisopropylethylamine (0.76 ml, 4.4 mmol) followed by diphenylchlorophosphate (0.91 ml, 4.1 mmol). After stirring the reaction mixture at room temperature for 1 hour, diisopropylethylamine (0.76 ml, 4.4 mmol) was added and then a solution of 2-methylthio-3-methyl-1-(2-mercaptoethyl)imidazolium trifluoromethanesulfonate (2) .0 g, 5.9 mmol) in 5 ml of acetonitrile. The reaction mixture was kept at room temperature for 10 hours and then concentrated in vacuo to obtain a gum. This was collected in H 2 O, then with 20* acetonitrile-H 2 O and finally 30* acetonitrile-H 2 O, followed by lyophilization of the appropriate fractions gave the product in an amount of 0.90 g corresponding to 30* as a pale yellow solid.
IR (KBr) vmaks: 3380 (br, OH), 1770 (e-laktam CO) cm-<1>; IR (KBr) vmax: 3380 (br, OH), 1770 (e-lactam CO) cm-<1>;
<1->H-NMR (d6-aceton) 5: 8,35 (br, s, 1H), 8,24, 7,78 (ABq, <1->H-NMR (d6-acetone) δ: 8.35 (br, s, 1H), 8.24, 7.78 (ABq,
J-8,8 Hz, 4H, aromatisk), 7,89 (br s, 1H), 7,25-6,91 (m, 10H, difenylfosfat), 5,50, 5,25 (ABq, J-12 Hz, 2H, benzylisk), 4,75-4,27 (m, 3H), 4,03 (s, 3H, N-Me), 4,15-2,75 (m, 8H), 2,53 (s, 3H, S-Me), 1,22 (d, J-6,2 Hz, 3H, -CHMe). J-8.8 Hz, 4H, aromatic), 7.89 (br s, 1H), 7.25-6.91 (m, 10H, diphenyl phosphate), 5.50, 5.25 (ABq, J-12 Hz, 2H, benzylic), 4.75-4.27 (m, 3H), 4.03 (s, 3H, N-Me), 4.15-2.75 (m, 8H), 2.53 ( s, 3H, S-Me), 1.22 (d, J-6.2 Hz, 3H, -CHMe).
C. ( 5R. 6S)- 3- f2-( 2- metvltio- 3- metvllmidazolio) etvltlo1- 6- ri-( R)- hvdroksvetvll- 7- okso- l- azabicvklo( 3. 2. 0) hept- 2- en- 2-karboksvlat C. ( 5R. 6S)- 3- f2-( 2- metvlthio- 3- metvllmidazolio) etvltlo1- ri-( R)- hvdroxvetvll- 7- oxo- 1- azabicclo( 3. 2. 0) hept- 2 - a- 2-carboxylate
Til en oppløsning av p-nitrobenzyl-(5R,6S)-3-[2-(2-metyltio-3-metylimidazolio)etyltlo]-6-[l-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat-dlfenylfosfat (1,20 g, 1,56 mmol) i en blanding av 70 ml THF, 70 ml eter og 31 ml fosfatbuffer (0,05 M, pH 7,4) ble det satt 1,2 g 10* palladium-på-trekull. Denne blanding ble hydrogenert (Parr) ved ca. 2,5 kp/cm<2> i 55 minutter. Reaksjonsblandingen ble derefter filtrert gjennom Celite, og filterkaken vasket med H2O og eter. Den vandige fase ble separert, avkjølt ved 0<*>C og pH-verdien innstilt til 7,0 med mettet vandig NaHC03. Efter å ha fjernet resterende organiske oppløsningsmidler under vakuum ble den vandige oppløsning satt til en C^g-reversfase-kolonne. Eluering med H2O og derefter 8* acetonitril-H20 og efterfølgende lyofilisering av de relevante fraksjoner ga 0,25 g av et fast stoff. Dette materialet ble renset igjen ved reversfase-HPLC under dannelse av produktet i en mengde av 0,114 g tilsvarende 19* som et off-white, fast stoff. To a solution of p-nitrobenzyl-(5R,6S)-3-[2-(2-methylthio-3-methylimidazolio)ethylthio]-6-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo (3.2.0)hept-2-ene-2-carboxylate-dlphenylphosphate (1.20 g, 1.56 mmol) in a mixture of 70 mL THF, 70 mL ether and 31 mL phosphate buffer (0.05 M, pH 7 ,4) 1.2 g of 10* palladium-on-charcoal was added. This mixture was hydrogenated (Parr) at approx. 2.5 kp/cm<2> for 55 minutes. The reaction mixture was then filtered through Celite, and the filter cake washed with H2O and ether. The aqueous phase was separated, cooled at 0<*>C and the pH adjusted to 7.0 with saturated aqueous NaHCO 3 . After removing residual organic solvents under vacuum, the aqueous solution was added to a C₂g reverse phase column. Elution with H 2 O and then 8* acetonitrile-H 2 O and subsequent lyophilization of the relevant fractions gave 0.25 g of a solid. This material was purified again by reverse phase HPLC to give the product in an amount of 0.114 g corresponding to 19* as an off-white solid.
IR (KBr) <v>maks: 3420 (OH), 1750 (e-laktam CO), 1590 (-C02<e>) IR (KBr) <v>max: 3420 (OH), 1750 (e-lactam CO), 1590 (-C02<e>)
cm<-1>; <i>H-NMR (D20) 5: 7,58 (s, 2H), 4,52 (t, J-6 Hz, 2H), 4,28-3,82 (m, 2H), 3,90 (s, 3H, n-Me), 3,40-2,87 (m, 5H), 2,40 (s, 3H, S-Me), 1,20 (d, J-6,4 Hz, 3H, —CHMe); cm<-1>; <i>H-NMR (D 2 O) δ: 7.58 (s, 2H), 4.52 (t, J-6 Hz, 2H), 4.28-3.82 (m, 2H), 3.90 (s, 3H, n-Me), 3.40-2.87 (m, 5H), 2.40 (s, 3H, S-Me), 1.20 (d, J-6.4 Hz, 3H , —CHMe);
UV (H20) Xmaks: 297 (c7572), 262 (c6259), 222 (c7955) nm. UV (H20) Xmax: 297 (c7572), 262 (c6259), 222 (c7955) nm.
Eksempel 12 Example 12
Fremstilling av ( 5R. 6S)- 3- r2-( 3- amlnopvridinlo/) etvltiol- 6- f 1-( R)- hvdroksyetvl1- 7- okso- l- azabicyklo( 3. 2. 0) hept- 2- en- 2-karboksvlat Preparation of (5R.6S)-3-r2-(3-amnolopvridinlo/)ethylthiol-6-f1-(R)-hydroxyethyl1-7-oxo-1-azabicyclo(3.2.0)hept-2-ene - 2-carboxylate
A. S- amlno- l- f 2- merkaptoetvl ) pyridinlnnikl nrl ri A. S-amnol-l- f 2- mercaptoetvl ) pyridinelnnicl nrl ri
3-aminopyridin (1,50 g, 0,16 mmol) ble oppsamlet i 15 ml IN metanollsk HC1 og den resulterende oppløsning inndampet inntil hydrokloridet var dannet som en olje. Til denne olje ble det satt 3-aminopyridin (1,32 g, 0,015 mmol) og etylensulfid (0,89 ml, 0,015 mmol), og den resulterende oppløsning ble oppvarmet i et oljebad ved 60-65<*>C under N2 i 2 timer. En ytterligere ekvivalent etylensulfid (0,89 ml, 0,015 mmol) ble tilsatt, og oppvarmingen fortsatt ved 55-65<*>C i 65 timer. Reaksjonsblandingen ble vasket med CH2CI2 og derefter samlet i 25 ml H2O. Den vandige oppløsning ble satt til en C^g-reversfase-kolonne, som ble eluert med H2O. Inndamping av de relevante fraksjoner ga produktet i en mengde på 1,26 g tilsvarende 44* som en farveløs, viskøs olje. 3-Aminopyridine (1.50 g, 0.16 mmol) was taken up in 15 mL of 1N methanolic HCl and the resulting solution evaporated until the hydrochloride formed as an oil. To this oil was added 3-aminopyridine (1.32 g, 0.015 mmol) and ethylene sulfide (0.89 mL, 0.015 mmol), and the resulting solution was heated in an oil bath at 60-65<*>C under N2 in 2 hours. An additional equivalent of ethylene sulfide (0.89 mL, 0.015 mmol) was added and heating continued at 55-65<*>C for 65 hours. The reaction mixture was washed with CH 2 Cl 2 and then taken up in 25 mL H 2 O. The aqueous solution was applied to a C 2 g reverse phase column, which was eluted with H 2 O. Evaporation of the relevant fractions gave the product in an amount of 1.26 g corresponding to 44* as a colorless, viscous oil.
IR (film) vmaks: 3180 (NH2) cm"<1>; IR (film) vmax: 3180 (NH2) cm"<1>;
1-H-NMR (DMS0-d6) 5: 8,19-7,59 (m, 4H, aromatisk), 4,59 (t, 1-H-NMR (DMS0-d6) δ: 8.19-7.59 (m, 4H, aromatic), 4.59 (t,
J-6,2 Hz, 2H, N-CH2), 3,5 (br s, 2H, -NH2), 3,20-2,77 (m, 3H). J-6.2 Hz, 2H, N-CH 2 ), 3.5 (br s, 2H, -NH 2 ), 3.20-2.77 (m, 3H).
B. p- nitrobenzvl-( 5R. 6S)- 3- r2- f3- aminopyridinio letyltlo"!- 6-l" l-( R )- hvdroksvetyll - 7- okso- l- azabicyklo( 3. 2. 0 ) hept- 2- en-2- karboksylat- difenylfosfat B. p-nitrobenzvl-(5R.6S)-3-r2-f3-aminopyridinioletyltlo"!-6-l"l-(R)-hydroxybutyl-7-oxo-l-azabicyclo(3.2.0)hept - 2- en-2- carboxylate diphenyl phosphate
Til en oppløsning av p-nitrobenzyl-(5R,6S)-6-[l-(R)-hydroksy-etyl] -3,7-diokso-l-azabicyklo(3.2.0 )heptan-2-karboksylat (0,696 g, 2,0 mmol) i 10 ml tørr acetonitril ved 0°C under N2, ble det dråpevis satt diisopropyletylamin (0,382 ml, 2,2 mmol) fulgt av difenylklorfosfat (0,457 ml, 2,2 mmol). Efter omrøring ved 0°C i 30 minutter ble det tilsatt en oppløsning av 3-amino-l-(2-merkaptoetyl)pyridiniumklorid (0,475 g, 2,5 mmol) i 1 ml tørr DMF, fulgt av ytterligere diisopropyletylamin (0,435 ml, 2,5 mmol). Reaksjonsblandingen ble holdt ved 0°C i VA time og derefter konsentrert under vakuum. Den resulterende gummi ble samlet i 1:1 acetonitril :H2<) og satt til C^g-reversfase-kolonne. Eluering med H20 fulgt av 20* acetonitril-H20 og efterfølgende lyofilisering av de relevante fraksjoner ga produktet i en mengde av 0,730 g, tilsvarende 50* som et beigefarvet, fast stoff. To a solution of p-nitrobenzyl-(5R,6S)-6-[l-(R)-hydroxy-ethyl]-3,7-dioxo-1-azabicyclo(3.2.0 )heptane-2-carboxylate (0.696 g , 2.0 mmol) in 10 mL of dry acetonitrile at 0°C under N 2 , diisopropylethylamine (0.382 mL, 2.2 mmol) was added dropwise followed by diphenylchlorophosphate (0.457 mL, 2.2 mmol). After stirring at 0°C for 30 minutes, a solution of 3-amino-1-(2-mercaptoethyl)pyridinium chloride (0.475 g, 2.5 mmol) in 1 mL of dry DMF was added, followed by additional diisopropylethylamine (0.435 mL, 2.5 mmol). The reaction mixture was kept at 0°C for VA hour and then concentrated under vacuum. The resulting gum was collected in 1:1 acetonitrile :H 2 <) and applied to a C 2 g reverse phase column. Elution with H 2 O followed by 20* acetonitrile-H 2 O and subsequent lyophilization of the relevant fractions gave the product in an amount of 0.730 g, corresponding to 50* as a beige solid.
IR (KBr) vmaks: 3330 (br, OH), 3180 (br, NH2), 1770 (e-laktam IR (KBr) vmax: 3330 (br, OH), 3180 (br, NH2), 1770 (e-lactam
CO), 1690 (-C02pNB) cm-<1>; CO), 1690 (-CO 2 pNB) cm-<1>;
<1->H-NMR (DMS0-d6) S: 8,29-7,63 (m, 8H, aromatisk), 7,2-6,7 <1->H-NMR (DMS0-d6) S: 8.29-7.63 (m, 8H, aromatic), 7.2-6.7
(m, 10H, difenylfosfat), 5,47, 5,18 (ABq, J-14 Hz, 2H, benzylisk), 4,73-4,45 (m, 3H), 4,2-3,8 (m, 1H), 3,6-2,6 (m, 8H), 1,15 (d, J-6,2 Hz, 3H, CHMe). (m, 10H, diphenyl phosphate), 5.47, 5.18 (ABq, J-14 Hz, 2H, benzylic), 4.73-4.45 (m, 3H), 4.2-3.8 (m , 1H), 3.6-2.6 (m, 8H), 1.15 (d, J-6.2 Hz, 3H, CHMe).
C. ( 5R. 6S)- 3-(( 2-( 3- aminopvridinioletvl) tlo)- 6- fl-( R)-hvdroksvetvll- 7- okso- l- azabicyklo( 3. 2. 0) hept- 2- en- 2-karboksvlat C. ( 5R. 6S )- 3-(( 2-( 3- aminopvridiniolethyl) tlo)- 6- fl-( R )-hydroxyl- 7- oxo- 1- azabicyclo( 3. 2. 0) hept- 2- a- 2-carboxylate
Til en blanding av p-nitrobenzyl-(5R,6S)-3-(2-(3-aminopyridinio)etyltio)-6-[l-(R)-hydroksyetyl]-7-okso-l-azabicyklo-(3.2.0)hept-2-en-2-karboksylat-difenylfosfat (0,730 g, 1,0 mmol) og 0,7 g 10* palladium-på-trekull i 25 ml fosfatbuffer (0,05 M, pH 7,4) ble det satt 8 ml THF og 20 ml eter. Denne blanding ble hydrogenert (Parr) ved ca. 2,8 kp/cm<2> i 1 time. Den resulterende blanding ble filtrert gjennom en pute av Celite og filterkaken ble vasket med H2O og eter. Den vandige fase ble separert, vasket to ganger med eter og derefter ble tilbakeblevne flyktige stoffer fjernet under vakuum. Den vandige oppløsning ble straks satt til en C^g-reversfase-kolonne som ble eluert med EtøO. Lyofilisering av de relevante fraksjoner ga 0,45 g av et off-white, fast stoff. Dette ble renset igjen ved reversfase-HPLC under dannelse av det ønskede produkt i en mengde av 0,123 g tilsvarende 35* som et elfenbensfarvet, fast stoff. To a mixture of p-nitrobenzyl-(5R,6S)-3-(2-(3-aminopyridinio)ethylthio)-6-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo-(3.2. 0)hept-2-ene-2-carboxylate diphenyl phosphate (0.730 g, 1.0 mmol) and 0.7 g of 10* palladium-on-charcoal in 25 ml of phosphate buffer (0.05 M, pH 7.4) were 8 ml of THF and 20 ml of ether were added. This mixture was hydrogenated (Parr) at approx. 2.8 kp/cm<2> for 1 hour. The resulting mixture was filtered through a pad of Celite and the filter cake was washed with H 2 O and ether. The aqueous phase was separated, washed twice with ether and then residual volatiles were removed under vacuum. The aqueous solution was immediately added to a C₂₂ reverse-phase column eluted with Et₂O. Lyophilization of the relevant fractions gave 0.45 g of an off-white solid. This was purified again by reverse phase HPLC to give the desired product in an amount of 0.123 g corresponding to 35* as an ivory colored solid.
IR (KBr) vmaks: 3340 (br), 1750 (br, e-laktam CO), 1580 (br, IR (KBr) vmax: 3340 (br), 1750 (br, e-lactam CO), 1580 (br,
-C02<9>) cm-<1>; <1->H-NMR (D20) S: 8,07-7,59 (m, 4H, aromatisk), 4,61 (t, J-5,8 Hz, 2H, N-CH2), 4,14 (d av q, J-J'-6,3 Hz, 2H, H-l'). 3,97 (d av t, J=9,2 Hz, J'=2,6 Hz, 1H, H-5), 3,38 (t, J-5,8 Hz, 2H, S-CH2), 3,24 (d av d, J-6,0 Hz, J'=2,6 Hz, 1H, H-6), 3,17-2,57 (m, 2H, H-4), 1,21 (d, J=6,3 Hz, 3H, CHMe); UV (H20) <X>maks: 299 (c7949), 256 (c8822) nm; t^ (pH 7,4, 36,8°C): 18,5 t. Eksempel 13 Fremstilling av -C02<9>) cm-<1>; <1->H-NMR (D 2 O) S: 8.07-7.59 (m, 4H, aromatic), 4.61 (t, J-5.8 Hz, 2H, N-CH 2 ), 4.14 (d of q, J-J'-6.3 Hz, 2H, H-1'). 3.97 (d of t, J=9.2 Hz, J'=2.6 Hz, 1H, H-5), 3.38 (t, J-5.8 Hz, 2H, S-CH2), 3.24 (d of d, J-6.0 Hz, J'=2.6 Hz, 1H, H-6), 3.17-2.57 (m, 2H, H-4), 1.21 (d, J=6.3 Hz, 3H, CHMe); UV (H 2 O) <X>max: 299 (c7949), 256 (c8822) nm; t^ (pH 7.4, 36.8°C): 18.5 h. Example 13 Preparation of
( 5R. 6S)- 3- r l-( S)- metvl- 2-( l- Dvrldiniiun) etvltiol- 6- r 1-( R)-hvdroksvetvll- 7- okso- l- azabicyklo( 3. 2. 0)hept-2-en-2-karboksvlat (5R.6S)-3-r1-(S)-methyl-2-(1-Diviridine)ethylthiol-6-r1-(R)-hydroxymethyl-7-oxo-1-azabicyclo(3.2.0 )hept-2-ene-2-carboxylate
og and
( 5R. 6S)- 3- ri-( R)- metvl- 2-( l- Pvridini\ un) etvltio1- 6- ri-( R)-hvdroksvetvll- 7- okso- l- azabicyklo( 3. 2 . 0 ) hept- 2- en- 2-karboksvlat ( 5R . 6S )- 3- ri-( R )- methyl- 2-( 1- Pvridini\un) ethvlthio1- 6- ri-( R )-hydroxymethyl- 7- oxo- 1- azabicyclo( 3. 2 . 0 ) hept-2-ene-2-carboxylate
A. dl- 1-( 2- merkapto- 2- metvletyl) pvrIri1ninmmetansulfonat dl- l-( 2- merkapto- l- metvletyl IpvridlriinmiTi fitansulfonat A. dl- 1-(2-mercapto-2-methylethyl)pyridinemethanesulfonate
Metansulfonsyre (1,95 ml, 0,030 mol) ble langsomt satt til kald pyridin (7,83 ml, 0,097 mol), og den resulterende blanding ble omrørt ved 40°C i 15 minutter, behandlet med d£-propylensulfid (2,59 ml, 0,033 mol) og omrørt ved 60°C under nitrogen i 90 timer. Pyridin ble fjernet under vakuum, resten ble blandet med vann og renset ved kromatografi (preparativ HPLC, Bondapak C^g). De passende fraksjoner ble forenet og lyofilisert under dannelse av dÆ-l-(2-merkapto-2-metyletyl)-pyridiniummetansulfonat, 1,14 g (15*) som en f arveløs sirup. Methanesulfonic acid (1.95 mL, 0.030 mol) was slowly added to cold pyridine (7.83 mL, 0.097 mol), and the resulting mixture was stirred at 40°C for 15 min, treated with d£-propylene sulfide (2.59 ml, 0.033 mol) and stirred at 60°C under nitrogen for 90 hours. Pyridine was removed under vacuum, the residue was mixed with water and purified by chromatography (preparative HPLC, Bondapak C^g). The appropriate fractions were combined and lyophilized to give dA-1-(2-mercapto-2-methylethyl)-pyridinium methanesulfonate, 1.14 g (15*) as a colorless syrup.
IR (film) vmaks: 2520 (SH), 1640 (pyrldinium), 1180 (s, IR (film) vmax: 2520 (SH), 1640 (pyrldinium), 1180 (s,
C<H>3S03<e>), 1040 (CH3S03<e>) cm-1; l-H-NMR (DMS0-d6) S: 1,35 (d, J-6,8 Hz, 3H, CH3CHS), 2,30 (s, 3H, CH3S03<e>), 2,90 (d, J-8,5 Hz, 1H, SH), 3,2-3,7 (m, CHSH), 4,52 (dd, <J>genr12'9 Hz» J-8,4 Hz, CHCH2N°), 4,87 (dd, Jgem-12,9 Hz, J-6,0 Hz, CHCH2<N>), 8,0-8,4 (m, 2H, Hm av pyrldinium), 8,5-8,8 (m, 1H, Hp av pyrldinium), 9,04 (dd, J-1,4 Hz, J-6,7 Hz, 2H, Ho av pyrldinium); C<H>3SO3<e>), 1040 (CH3SO3<e>) cm-1; 1-H-NMR (DMS0-d6) S: 1.35 (d, J-6.8 Hz, 3H, CH3CHS), 2.30 (s, 3H, CH3SO3<e>), 2.90 (d, J- 8.5 Hz, 1H, SH), 3.2-3.7 (m, CHSH), 4.52 (dd, <J>genr12'9 Hz» J-8.4 Hz, CHCH2N°), 4, 87 (dd, Jgem-12.9 Hz, J-6.0 Hz, CHCH2<N>), 8.0-8.4 (m, 2H, Hm of pyrldinium), 8.5-8.8 (m , 1H, Hp of pyrldinium), 9.04 (dd, J-1.4 Hz, J-6.7 Hz, 2H, Ho of pyrldinium);
UV (H20) Xmaks: 208 (c5267), 259 (c3338) nm; UV (H 2 O) Xmax: 208 (c5267), 259 (c3338) nm;
Analyse for CgH15N03S2•2H20: Analysis for CgH15N03S2•2H20:
Beregnet: C 37,88 H 6,71 N 4,91 S 22,47 Calculated: C 37.88 H 6.71 N 4.91 S 22.47
Funnet: C 37,49 H 6,85 N 4,86 S 22,09 Found: C 37.49 H 6.85 N 4.86 S 22.09
og d£-l-(2-merkapto-l-metyletyl)pyridiniummetansulfonat, 0,82 g (11*) som en farveløs sirup; and d£-1-(2-mercapto-1-methylethyl)pyridinium methanesulfonate, 0.82 g (11*) as a colorless syrup;
IR (film) vmaks: 2500 (SH), 1628 (pyrldinium), 1180 (sulfonat), 1035 (sulfonat) cm-<1>; IR (film) vmax: 2500 (SH), 1628 (pyrrldinium), 1180 (sulfonate), 1035 (sulfonate) cm-<1>;
<i>H-NMR (DMS0-d6) 5: 1,69 (d, J-6,8 Hz, 3H, CH3CHN<®>), 2,32 <i>H-NMR (DMS0-d6) δ: 1.69 (d, J-6.8 Hz, 3H, CH3CHN<®>), 2.32
(s, 3H, CH3S03<e>), 3,0-3,3 (m, 2H, CH2S), 4,2-5,2 (m, 1H, CHN<®>), 8,0-8,4 (m, 2H, Hm av pyrldinium), (s, 3H, CH3SO3<e>), 3.0-3.3 (m, 2H, CH2S), 4.2-5.2 (m, 1H, CHN<®>), 8.0-8, 4 (m, 2H, Hm of pyrldinium),
8,5-8,8 (m, 1H, Hp av pyrldinium), 9,0-9,2 (m, 2H, Ho av pyrldinium); 8.5-8.8 (m, 1H, Hp of pyrldinium), 9.0-9.2 (m, 2H, Ho of pyrldinium);
UV (H20) Xmaks: 209 (c4987), 158 (c3838) nm; UV (H 2 O) Xmax: 209 (c4987), 158 (c3838) nm;
Analyse for C9H15NO3S2* 1,5H20: Analysis for C9H15NO3S2* 1.5H20:
Beregnet: C 39,11 H 6,56 N 5,07 Calculated: C 39.11 H 6.56 N 5.07
Funnet: C 39,11 H 5,92 N 5,20 Found: C 39.11 H 5.92 N 5.20
B. ( 5R. 6S)- p- nltrobenzvl- 3- r1- fR. S)- metvi- 2-( 1- pvridinlum)-etvltiol - 6- l" l-( R )- hydroksyetvl" l - 7- okso- l- azablcvklo-( 3. 2. 0 ) hept- 2- en- 2- karboksylat- difenylfosfat B. ( 5R. 6S)- p- nltrobenzvl- 3- r1- fR. S)- metvi- 2-( 1- pvridinlum)-ethylthiol - 6- l" l-( R )- hydroxyethyl" l - 7- oxo- l- azablciclo-( 3. 2. 0 ) hept- 2- en- 2- carboxylate diphenyl phosphate
Til en kald (0°C) oppløsning av (5R,6S)-p-nitrobenzyl-6-[1-(R)-hydroksyetyl]-3,7-diokso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat (0,523 g, 1,5 mmol) i 6 ml acetonitril, holdt under nitrogenatmosfære, ble det satt diisopropyletylamin (0,314 ml, 1,8 mmol) fulgt av difenylklorfosfat (0,373 ml, 1,8 mmol). Reaksjonsblandingen ble omrørt i 30 minutter og behandlet med en oppløsning av d£-l-(2-merkapto-2-metyletyl)-pyridiniummetansulfonat (0,539 g, 2,16 mmol) i 2 ml acetonitril og diisopropyletylamin (0,314 ml, 1,8 mmol). Reaksjonsblandingen ble omrørt ved 0°C i 1 time, fortynnet med 24 ml kaldt vann av 0°C og kromatografert over en 2,5 x 8,5 cm preparatlv Bondapak C18-kolonne med 25-50* acetonitril i vann som elueringsoppløsningsmidler under dannelse av 1,07 g tilsvarende 97* av tittelforbindelsen som et gulaktig pulver efter lyofilisering; To a cold (0°C) solution of (5R,6S)-p-nitrobenzyl-6-[1-(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)hept-2- ene-2-carboxylate (0.523 g, 1.5 mmol) in 6 mL of acetonitrile, kept under a nitrogen atmosphere, was added diisopropylethylamine (0.314 mL, 1.8 mmol) followed by diphenylchlorophosphate (0.373 mL, 1.8 mmol). The reaction mixture was stirred for 30 min and treated with a solution of d£-1-(2-mercapto-2-methylethyl)-pyridinium methanesulfonate (0.539 g, 2.16 mmol) in 2 mL of acetonitrile and diisopropylethylamine (0.314 mL, 1.8 mmol). The reaction mixture was stirred at 0°C for 1 hour, diluted with 24 ml of cold 0°C water and chromatographed over a 2.5 x 8.5 cm prepared Bondapak C18 column with 25-50% acetonitrile in water as elution solvents during formation of 1.07 g corresponding to 97% of the title compound as a yellowish powder after lyophilization;
IR (KBr) vmaks: 3700-3100 (OH), 1770 (C-0 av e-laktam), 1695 IR (KBr) vmax: 3700-3100 (OH), 1770 (C-0 of ε-lactam), 1695
(C-0 av pNB-ester), 1630 (pyrldinium), 1590 (fenyl), 1518 (N02), 1348 (N02), 885 (N02) cm-<1>; (C-0 of pNB ester), 1630 (pyrldinium), 1590 (phenyl), 1518 (NO2), 1348 (NO2), 885 (NO2) cm-<1>;
1-H-NMR (DMS0-d6) 5: 1,14 (d, J-6,1 Hz, 3H, CH3CH0), 133 (d, 1-H-NMR (DMSO-d6) δ: 1.14 (d, J-6.1 Hz, 3H, CH3CH0), 133 (d,
J-6,3 Hz, 3H, CH3CHS), 4,6-5,0 (m, CH2N<8>), 5,14 (d, J=5,2 Hz, 1H, OH), 5,37 (sentrum av ABq, J-12,4 Hz, 3H, CH2 av pNB), 6,6-7,5 (m, 10H, fenyl av fosfat), 7,69 (d, J-8,7 Hz, 2H, Ho av pNB), 8,0-8,4 (m, 4H; Hm av pNB, Hm av pyrldinium), 8,4-8,8 (m, 1H, Hp av pyrldinium), 9,08 (d, J-5,6 Hz, 2H, Ho av pyrldinium ); J-6.3 Hz, 3H, CH3CHS), 4.6-5.0 (m, CH2N<8>), 5.14 (d, J=5.2 Hz, 1H, OH), 5.37 ( center of ABq, J-12.4 Hz, 3H, CH2 of pNB), 6.6-7.5 (m, 10H, phenyl of phosphate), 7.69 (d, J-8.7 Hz, 2H, Ho of pNB), 8.0-8.4 (m, 4H; Hm of pNB, Hm of pyrldinium), 8.4-8.8 (m, 1H, Hp of pyrldinium), 9.08 (d, J -5.6 Hz, 2H, Ho of pyrldinium );
UV (H20) <X>maks: 263 (cl3325), 308 (c8915) nm. UV (H20) <X>max: 263 (cl3325), 308 (c8915) nm.
Analyse for C36<H>36<N>3<0>10SP'H20: Analysis for C36<H>36<N>3<0>10SP'H20:
Beregnet: C 57,52 H 5,10 N 5,59 S 4,27 Funnet: C 57,76 H 4,96 N 5,36 S 4,35 C. ( 5R. 6S)- 3- ri-( R og S)- metvl- 2-( l- pvridinium) etvltio1- 6- ri- ( R )- hvdroksyetyl" l - 7- okso- l- azabicyklo( 3. 2. 0) hept- 2- en- 2-karboksylat Calculated: C 57.52 H 5.10 N 5.59 S 4.27 Found: C 57.76 H 4.96 N 5.36 S 4.35 C. ( 5R. 6S)- 3- ri-( R and S)-methyl-2-(1-pvridinium)ethylthiol-6-ri- ( R )- hydroxyethyl" 1 - 7- oxo- 1- azabicyclo( 3. 2. 0 ) hept- 2-ene- 2-carboxylate
Til en oppløsning av (5R,6S)-p-nitrobenzyl-3-[l-(R,S)-metyl-2-(1-pyridinium )etyltio]-6-[1-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0 )hept-2-en-2-karboksylat-difenylfosfat (0,60 g, 0,82 mmol) i 33 ml våt tetrahydrofuran ble det satt 33 ml eter, monobasisk kaliumfosfat-natriumhydroksydbuffer (17 ml, 0,15 M, pH 7,22) og 0,60 g 10* palladium-på-trekull. Den resulterende blanding ble hydrogenert i 1 time under ca. 2,8 kp/cm<2> ved 23"C. De to sjikt ble separert og det organiske sjiktet ekstrahert med 3 x 7 ml vann. De vandige sjikt ble forenet, filtrert gjennom en Celite-pute, vasket med 3 x 20 ml eter og kromatografert på en 2,5 x 9,5 cm preparativ Bondapak C^g-kolonne med vann som elueringsmiddel, hvorved man oppnådde 0,18 g tilsvarende 63* av en blanding av diastereoisomerer. Disse ble separert ved hjelp av HPLC (preparativ Bondapak C^g) med vann som elueringsoppløsnings-middel; To a solution of (5R,6S)-p-nitrobenzyl-3-[1-(R,S)-methyl-2-(1-pyridinium )ethylthio]-6-[1-(R)-hydroxyethyl]-7 -oxo-1-azabicyclo(3.2.0 )hept-2-ene-2-carboxylate-diphenylphosphate (0.60 g, 0.82 mmol) in 33 ml of wet tetrahydrofuran was added 33 ml of ether, monobasic potassium phosphate-sodium hydroxide buffer ( 17 ml, 0.15 M, pH 7.22) and 0.60 g of 10* palladium-on-charcoal. The resulting mixture was hydrogenated for 1 hour under approx. 2.8 kp/cm<2> at 23"C. The two layers were separated and the organic layer extracted with 3 x 7 ml of water. The aqueous layers were combined, filtered through a Celite pad, washed with 3 x 20 ml ether and chromatographed on a 2.5 x 9.5 cm preparative Bondapak C^g column with water as eluent, whereby 0.18 g corresponding to 63* of a mixture of diastereoisomers was obtained. These were separated by means of HPLC (preparative Bondapak C^g) with water as elution solvent;
isomer med lavere retensjonstid, 0,068 g (23*), forbindelse lower retention isomer, 0.068 g (23*), compound
"B"; "B";
IR (KBr) vmaks: 1770 (C=0 av p<->laktam), 1633 (pyrldinium), IR (KBr) vmax: 1770 (C=0 of p<->lactam), 1633 (pyrldinium),
1593 (karboksylat) cm-<1>; 1593 (carboxylate) cm-<1>;
%-NMR (D20) S: 1,20 (d, J-6,3 Hz, 3H, CH3CH0), 1,42 (d, %-NMR (D 2 O) S: 1.20 (d, J-6.3 Hz, 3H, CH 3 CH 0 ), 1.42 (d,
J=6,9 Hz, 3H, CH3CHS), 2,3-3,2 (m, 3H, H-4, H-6), 3,5-3,9 (m, 1H, SCH), 3,9-4,2 (m, 2H, H-5, CH3CHO), 4,3-5,1 (m, CH2N<®>), 7,8-8,2 (m, 2H, Hm av pyrldinium), 8,4-8,7 (m, 1H, Hp av pyrldinium), 8,7-9,0 (m, 2H, Ho av pyrldinium); J=6.9 Hz, 3H, CH3CHS), 2.3-3.2 (m, 3H, H-4, H-6), 3.5-3.9 (m, 1H, SCH), 3, 9-4.2 (m, 2H, H-5, CH3CHO), 4.3-5.1 (m, CH2N<®>), 7.8-8.2 (m, 2H, Hm of pyrldinium), 8.4-8.7 (m, 1H, Hp of pyrldinium), 8.7-9.0 (m, 2H, Ho of pyrldinium);
UV (H20) Xmaks: 260 (c6727), 300 (c8245) nm; UV (H2O) Xmax: 260 (c6727), 300 (c8245) nm;
[a]<g3> -39,3<*> (c, H20); [a]<g3> -39.3<*> (c, H2O);
t^ - 12,6 timer (målt ved en konsentrasjon på IO"<4> M i t^ - 12.6 hours (measured at a concentration of 10"<4> M i
fosfatbuffer, pH 7,4 ved 36,8<*>C); phosphate buffer, pH 7.4 at 36.8<*>C);
isomer med høyere retensjonstid, 0,081 g (28*), forbindelse isomer with higher retention time, 0.081 g (28*), compd
"A"; "A";
IR (KBr) vmaks: 1755 (C-0 av e-laktam), 1630 (pyrldinium), IR (KBr) vmax: 1755 (C-0 of ε-lactam), 1630 (pyrldinium),
1590 (karboksylat) cm-<1>; 1590 (carboxylate) cm-<1>;
<*>H-NMR (D20) S: 1,18 (d, J-6,3 Hz, 3H, CH3CH0), 1,40 (d, J-7,0 Hz, 3H, CH3CHS), 2,84 (d, J-9,3 Hz, 2H, H-4), 3,26 (dd, J-2,7 Hz, J-5,9 Hz, 1H, H-6), 3,4-4,2 (m, 3H, SCH, CH3CHO, H-5), 4,2-5,1 (m, CH2N<®>), 7,7-8,1 (m, 2H, Hm av pyrldinium), 8,3-8,65 (m, 1H, Hp av pyrldinium), 8,65-8,9 (m, 2H, Ho av pyrldinium); <*>H-NMR (D20) S: 1.18 (d, J-6.3 Hz, 3H, CH3CH0), 1.40 (d, J-7.0 Hz, 3H, CH3CHS), 2.84 (d, J-9.3 Hz, 2H, H-4), 3.26 (dd, J-2.7 Hz, J-5.9 Hz, 1H, H-6), 3.4-4, 2 (m, 3H, SCH, CH3CHO, H-5), 4.2-5.1 (m, CH2N<®>), 7.7-8.1 (m, 2H, Hm of pyrldinium), 8, 3-8.65 (m, 1H, Hp of pyrldinium), 8.65-8.9 (m, 2H, Ho of pyrldinium);
UV (H20) Xmaks: 259 (c5694), 296 (c6936) nm; UV (H 2 O) Xmax: 259 (c5694), 296 (c6936) nm;
[a]2,3 +96,9° (c 0,56, H20); [α]2.3 +96.9° (c 0.56, H 2 O);
ttø = 15,6 timer (målt ved en konsentrasjon på 10~<4> M i ttø = 15.6 hours (measured at a concentration of 10~<4> M i
fosfatbuffer, pH 7,4 ved 36,8'C). phosphate buffer, pH 7.4 at 36.8'C).
Eksempel 14 Example 14
Fremstilling av Manufacture of
( 5R. 6S)- 3- r2- r( S)-( l- pvrldlniumn- l-( S)- cvkloheksvltlol- 6- ri-( R )- hydroksyetyl" l - 7- okso- l- azabicvklo( 3. 2. 0 ) hept- 2- en- 2-karboksvlat ( 5R. 6S)- 3- r2- r( S)-( l- pvrldlniumn- l-( S)- cvclohexvltlol- 6- r-( R )- hydroxyethyl" l - 7- oxo- l- azabicvclo( 3. 2. 0 ) hept- 2- en- 2-carboxvlat
og and
( 5R. 6S)- 3- f2- r( R)-( l- pvrldlniumn- l-( R)- cvkloheksvltlo1- 6- ri-( R )- hvdroksyetyl" l - 7- okso- l- azablcyklo( 3. 2. 0) hept- 2- en- 2-karboksylat ( 5R. 6S)- 3- f2- r( R)-( l- pvrldlniumn- l-( R)- cvclohexvltilo1- 6- ri-( R )- hydroxyethyl" l - 7- oxo- l- azablcyclo( 3. 2. 0) hept-2-ene-2-carboxylate
A. dÆ- l- r2- merkapto- l- cykloheksyll<py>rjdininmmetansulfonatA. dÆ-l-r2-mercaptol-l-cyclohexyl<py>rjdininmethanesulfonate
Metansulfonsyre (0,65 ml, 0,01 mol) ble dråpevis satt til pyridin (2,42 ml, 0,03 mol) under avkjøling. Blandingen ble omrørt under nitrogen i 10 minutter, behandlet med d£-cykloheksensulfid 2,377 g (85* rent), 0,0103 mol og omrørt ved 72"C i 25 timer. Overskuddet av pyridin ble fjernet under vakuum og sporene samdest!llert med vann. Resten ble blandet med vann og kromatografert på en 5 x 13 cm preparativ Bondapak C^g-kolonne med 0-2* acetonitril i vann som elueringsmiddel, hvorved det efter lyofilisering ble dannet en farveløs sirup, 1,57 g (53*); Methanesulfonic acid (0.65 mL, 0.01 mol) was added dropwise to pyridine (2.42 mL, 0.03 mol) with cooling. The mixture was stirred under nitrogen for 10 minutes, treated with d£-cyclohexene sulfide 2.377 g (85% pure), 0.0103 mol and stirred at 72°C for 25 hours. The excess pyridine was removed under vacuum and the traces co-distilled with water. The residue was mixed with water and chromatographed on a 5 x 13 cm preparative Bondapak C^g column with 0-2* acetonitrile in water as eluent, whereby after lyophilization a colorless syrup was formed, 1.57 g (53* );
IR (film) vmaks: 2500 (SH), 1625 (pyrldinium), 1190 (S03<G>)<;>IR (film) vmax: 2500 (SH), 1625 (pyrrldinium), 1190 (S03<G>)<;>
^H-NMR (DMSO-d6) S: 1,2-2,5 (m, 8H, cykloheksyl H), 2,32 (s, ^H-NMR (DMSO-d6) S: 1.2-2.5 (m, 8H, cyclohexyl H), 2.32 (s,
3H, CH3S03<e>), 2,82 (d, J-9,8 Hz, SH), 3,0-3,5 (m, 1H, CHSH), 4,2-4,9 (m, 1H, CHN<®>), 8,0-8,3 (m, 2H, Hm av pyridinium), 8,4-8,8 (m, 1H, Hp av pyrldinium), 8,9-9,3 (m, 2H, Ho av pyridinium); 3H, CH3SO3<e>), 2.82 (d, J-9.8 Hz, SH), 3.0-3.5 (m, 1H, CHSH), 4.2-4.9 (m, 1H , CHN<®>), 8.0-8.3 (m, 2H, Hm of pyridinium), 8.4-8.8 (m, 1H, Hp of pyrldinium), 8.9-9.3 (m , 2H, Ho of pyridinium);
UV (H20) Xmaks: 214 (c5365), 258 (c3500) nm; UV (H2O) Xmax: 214 (c5365), 258 (c3500) nm;
Analyse for C12H1gN03S2"H20: Analysis for C12H1gN03S2"H20:
Beregnet: C 46,88 H 6,88 N 4,56 Calculated: C 46.88 H 6.88 N 4.56
Funnet: C 46,61 H 6,46 N 4,65 Found: C 46.61 H 6.46 N 4.65
B. ( 5R. 6S)- p- nitrobenzvl- 3- f2- f( R eller S)-( 1- pvridinium) H-( R eller S)- cvkloheksvltlo1- 6- fl-( R)- hvdroksvetvl1- 7-okso- l- azabicyklo( 3. 2. 0) hept- 2- en- 2- karboksvlat- difenylfosfat B. ( 5R. 6S)- p- nitrobenzvl- 3- f2- f( R or S)-( 1- pvridinium) H-( R or S)- cvclohexvlto1- 6- fl-( R)- hvdroxvevl1- 7- oxol- l- azabicyclo( 3. 2. 0) hept- 2- en- 2- carboxyvlate- diphenyl phosphate
Til en kald (0°C) oppløsning av (5R,6S)-p-nitrobenzyl-6-[l-(R)-hydroksyetyl]-3,7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat (1,37 g, 3,93 mmol) i 15 ml acetonitril, holdt under nitrogen, ble det satt diisopropyletylamin (0,822 ml, 4,7 mmol) og difenylklorfosfat (0,979 ml, 4,7 mmol). Den resulterende oppløsning ble omrørt i 30 minutter og behandlet med en oppløsning av d£-l-(2-merkapto-l-cykloheksyl)-pyridiniummetansulfonat (1,64 g, 5,66 mmol) i 4,7 ml acetonitril, fulgt av diisopropyletylamin (0,822 ml, 4,7 mmol). Reaksjonsblandingen ble omrørt ved 0<*>C i 1 time, fortynnet med 75 ml kaldt vann av 0<*>C og kromatografert på preparativ Bondapak C^g med 25-50* acetonitril i vann som elueringsoppløsningsmiddel, hvorved det efter lyofilisering av de passende fraksjoner ble dannet 1,9 g tilsvarende 53* av tittelforbindelsen. To a cold (0°C) solution of (5R,6S)-p-nitrobenzyl-6-[l-(R)-hydroxyethyl]-3,7-oxo-1-azabicyclo(3.2.0)hept-2- ene-2-carboxylate (1.37 g, 3.93 mmol) in 15 mL of acetonitrile, kept under nitrogen, was added diisopropylethylamine (0.822 mL, 4.7 mmol) and diphenylchlorophosphate (0.979 mL, 4.7 mmol). The resulting solution was stirred for 30 min and treated with a solution of δ-1-(2-mercapto-1-cyclohexyl)-pyridinium methanesulfonate (1.64 g, 5.66 mmol) in 4.7 mL of acetonitrile, followed by diisopropylethylamine (0.822 mL, 4.7 mmol). The reaction mixture was stirred at 0<*>C for 1 hour, diluted with 75 ml of cold water at 0<*>C and chromatographed on preparative Bondapak C^g with 25-50* acetonitrile in water as elution solvent, whereby after lyophilization of the appropriate fractions were formed 1.9 g corresponding to 53* of the title compound.
IR (KBr) vmaks: 3700-3000 (0H), 1770 (C-0 av 3-laktam), 1700 IR (KBr) vmax: 3700-3000 (0H), 1770 (C-0 of 3-lactam), 1700
(C=0 av pNB-ester), 1628 (pyridinium), 1590 (fenyl), 1515 (N02), 1345 (N02), 880 (N02) cm-<1>; (C=0 of pNB ester), 1628 (pyridinium), 1590 (phenyl), 1515 (NO 2 ), 1345 (NO 2 ), 880 (NO 2 ) cm-<1>;
<1->H-NMR (D20) 5: 1,13 (d, J-6,1 Hz, 3H, CH3CH0), 1,2-2,5 (m, <1->H-NMR (D2O) δ: 1.13 (d, J-6.1 Hz, 3H, CH3CH0), 1.2-2.5 (m,
8H, cykloheksyl H), 2,7-3,5 (m, 4H, H-4, H-6, CHS), 3,5-4,4 (m, 2H, CH3CHO, H-5), 4,4-5,0 (m, 1H, CHN<®>), 5,30 (sentrum av ABq, J=12,8 Hz, CH2 av pNB), 6,7-7,4 (m, 10H, fenyl), 7,65 (d, J-8,6 Hz, 2H, Ho av pNB), 7,9-8,4 (m, 4H, Hm av pNB, Hm av pyridinium), 8,4-8,8 (m, 1H, Hp av pyridinium), 9,0-9,4 (m, 2H, Ho av pyridinium); 8H, cyclohexyl H), 2.7-3.5 (m, 4H, H-4, H-6, CHS), 3.5-4.4 (m, 2H, CH3CHO, H-5), 4, 4-5.0 (m, 1H, CHN<®>), 5.30 (center of ABq, J=12.8 Hz, CH2 of pNB), 6.7-7.4 (m, 10H, phenyl) , 7.65 (d, J-8.6 Hz, 2H, Ho of pNB), 7.9-8.4 (m, 4H, Hm of pNB, Hm of pyridinium), 8.4-8.8 ( m, 1H, Hp of pyridinium), 9.0-9.4 (m, 2H, Ho of pyridinium);
UV (H20) <X>maks: 263 (c9038), 309 (c6394) nm; UV (H 2 O) <X>max: 263 (c9038), 309 (c6394) nm;
Analyse for C3g<H>40<N>3010SP-H20: Analysis for C3g<H>40<N>3010SP-H20:
Beregnet: C 59,16 H 5,35 N 5,31 Calculated: C 59.16 H 5.35 N 5.31
Funnet: C 58,95 H 5,15 N 5,57 Found: C 58.95 H 5.15 N 5.57
C. ( 5R. 6S)- 3- f2-( R eller S)-( l- pvrldlniumn- l-( R eller Sl-cvkloheksvltlol- 6- l" l-( R)- hvdroksvetvl1- 7- okso- l- aza-blcyklo( 3. 2. 0 ) hept- 2- en- 2- karboksvlat C. ( 5R. 6S)- 3- f2-( R or S)-( l- pvrldlniumn- l-( R or Sl- cvclohexvollol- 6- l" l-( R)- hydroxyvetll1- 7- oxo- l- aza-blcyclo( 3. 2. 0 ) hept- 2- en- 2- carboxylate
Til en oppløsning av (5R,6S)-p-nitrobenzyl-3-[2-[(R eller S)-(l-pyridinium)-l-(R eller S)-cykloheksyltio]-6-[l-(R)-hydroksyetyl]-7-okso-l-azablcyklo(3.2.0)hept-2-en-2-karboksylat-difenylfosfat (1,85 g, 2,34 mmol) i 96 ml våt tetrahydrofuran ble det satt 96 ml eter, monobasisk kaliumfosfat-natriumhydroksydbuffer (0,15 M, pH 7,22, 50 ml) og 1,9 g 10* palladium-på-trekull. Den resulterende blanding ble hydrogenert ved 23°C under ca. 2,8 kp/cm<2> 1 1,25 timer. Det organiske sjikt ble separert og vasket med 3 x 25 ml vann. De vandige oppløsninger ble filtrert gjennom en Celite-pute, vasket med 2 x 60 ml eter, pumpet for å fjerne spor av organiske oppløsningsmidler og kromatografert på en 4,5 x 9 cm preparativ Bondapak Cig-kolonne med 0-5* acetonitril i vann som elueringsmiddel, hvorved det efter lyofilisering ble dannet 0,705 g tilsvarende 76* av en blanding av diastereoisomerer. Separering av disse ble foretatt ved hjelp av HPLC med 4* acetonitril i vann som elueringsmiddel; diastereoisomerer med lavere retensjonstid, forbindelse "A", (0,29 g, 31*); To a solution of (5R,6S)-p-nitrobenzyl-3-[2-[(R or S)-(l-pyridinium)-1-(R or S)-cyclohexylthio]-6-[l-(R )-hydroxyethyl]-7-oxo-1-azablcyclo(3.2.0)hept-2-ene-2-carboxylate diphenylphosphate (1.85 g, 2.34 mmol) in 96 ml of wet tetrahydrofuran was added to 96 ml of ether , monobasic potassium phosphate-sodium hydroxide buffer (0.15 M, pH 7.22, 50 ml) and 1.9 g of 10* palladium-on-charcoal. The resulting mixture was hydrogenated at 23°C under approx. 2.8 kp/cm<2> 1 1.25 hours. The organic layer was separated and washed with 3 x 25 ml of water. The aqueous solutions were filtered through a Celite pad, washed with 2 x 60 ml ether, pumped to remove traces of organic solvents and chromatographed on a 4.5 x 9 cm preparative Bondapak Cig column with 0-5* acetonitrile in water as eluent, whereby after lyophilization 0.705 g corresponding to 76* of a mixture of diastereoisomers was formed. Separation of these was carried out using HPLC with 4* acetonitrile in water as eluent; lower retention diastereoisomers, compound "A", (0.29 g, 31*);
IR (KBr) vmaks: 1750 (C-0 av e-laktam), 1620 (sh, pyridinium), 1685 (karboksylat) cm-<1>; IR (KBr) vmax: 1750 (C-0 of ε-lactam), 1620 (sh, pyridinium), 1685 (carboxylate) cm-<1>;
^H-NMR (D20) S: 1,21 (d, J-6,3 Hz, 3H, CH3CH0), 1,4-2,5 (m, ^H-NMR (D2O) S: 1.21 (d, J-6.3 Hz, 3H, CH3CH0), 1.4-2.5 (m,
8H, cykloheksyl H), 2,5-3,05 (m, 2H, H-4), 3,05-3,25 (m, 1H, H-6), 3,3-3,7 (m, 1H, CHS), 3,9-4,3 (m, 2H, H-5, CH3CHO), 4,3-4,8 (m, CHN<®>), 7,8-8,2 (m, 2H, Hm av pyridinium), 8,3-8,7 (m, 1H, Hp av pyridinium), 8,8-9,1 (m, 2H, Ho av pyridinium); 8H, cyclohexyl H), 2.5-3.05 (m, 2H, H-4), 3.05-3.25 (m, 1H, H-6), 3.3-3.7 (m, 1H, CHS), 3.9-4.3 (m, 2H, H-5, CH3CHO), 4.3-4.8 (m, CHN<®>), 7.8-8.2 (m, 2H, Hm of pyridinium), 8.3-8.7 (m, 1H, Hp of pyridinium), 8.8-9.1 (m, 2H, Ho of pyridinium);
UV (H20) Xmal£S: 260 (c7123), 300 (c8685) nm; UV (H 2 O) Xmal£S: 260 (c7123), 300 (c8685) nm;
[a]g3 +6,2' (c 0,63, H20); [α]g 3 +6.2' (c 0.63, H 2 O);
ttø - 16,6 timer (målt ved en konsentrasjon på 10~<4> M i ttø - 16.6 hours (measured at a concentration of 10~<4> M i
fosfatbuffer, pH 7,4 ved 36,8<*>C); phosphate buffer, pH 7.4 at 36.8<*>C);
Analyse for C2oH24<N>2<0>4S<<>2H20: Analysis for C2oH24<N>2<0>4S<<>2H20:
Beregnet: C 56,59 H 6,65 N 6,60 S 7,55 Calculated: C 56.59 H 6.65 N 6.60 S 7.55
Funnet: C 56,83 H 6,47 N 6,59 S 7,43; Found: C 56.83 H 6.47 N 6.59 S 7.43;
diastereoisomerer med høyere retensjonstid, forbindelse "B", (0,35 g, 38*); higher retention diastereoisomers, compound "B", (0.35 g, 38*);
IR (KBr) vmaks: 1750 (C-0 av e-laktam), 1622 (sh, pyridinium), 1588 (karboksylat) cm-<1>; IR (KBr) vmax: 1750 (C-0 of ε-lactam), 1622 (sh, pyridinium), 1588 (carboxylate) cm-<1>;
<i>H-NMR (D20) S: 1,19 (d, J-6,4 Hz, 3H, CH3CHO), 1,3-2,5 (m, <i>H-NMR (D2O) S: 1.19 (d, J-6.4 Hz, 3H, CH3CHO), 1.3-2.5 (m,
8H, cykloheksyl H), 2,5-3,1 (m, 2H, H-4), 3,1-3,3 (m, 1H, H-6), 3,3-3,8 (m, 2H, H-5, CHS), 4,1 (sentrum av m, 1H, CH3CHO), 4,25-4,7 (m, 1H, CHN<®>), 7,8-8,1 (m, 2H, Hm av pyridinium), 8,3-8,7 (m, 1H, Hp av pyridinium), 8,75-9,0 (m, 2H, Ho av pyridinium); 8H, cyclohexyl H), 2.5-3.1 (m, 2H, H-4), 3.1-3.3 (m, 1H, H-6), 3.3-3.8 (m, 2H, H-5, CHS), 4.1 (center of m, 1H, CH3CHO), 4.25-4.7 (m, 1H, CHN<®>), 7.8-8.1 (m, 2H, Hm of pyridinium), 8.3-8.7 (m, 1H, Hp of pyridinium), 8.75-9.0 (m, 2H, Ho of pyridinium);
UV (H20) Xmaks: 259 (c5992), 296 (c7646) nm; UV (H 2 O) Xmax: 259 (c5992), 296 (c7646) nm;
[a]<g3> +65,3° (c 0,43, H20); [α]<g3> +65.3° (c 0.43, H 2 O);
ty - 20,2 timer (målt ved en konsentrasjon på 10~<4> M i ty - 20.2 hours (measured at a concentration of 10~<4> M i
fosfatbuffer, pH 7,4 ved 36,8<*>C); phosphate buffer, pH 7.4 at 36.8<*>C);
Eksempel 15 Example 15
A. ( 5R)- allvl- 3- rf2- Pvrldinioetvl) tlo1-( 6S)- ri-( R)- hvdroksvetvll- 7- okso- l- azabicvklo( 3. 2. 0) hept- 2- en- 2- karboksvlat-difenvlfosfat A. ( 5R)- allvl- 3- rf2- Pvrldinioetvl) tlo1-( 6S)- ri-( R)- hvdroksvetvll- 7- oxo- l- azabicvklo( 3. 2. 0) hept- 2- en- 2- carboxyvlate-diphenyl phosphate
Til en oppløsning av (5R)-allyl-3,7-diokso-(6S)-[l-(R)-hydroksyetyl]-l-azabicyklo(3.2.0)heptan-2-(2R)-karboksylat (473 mg, 1,87 mmol) i CH3CN (6 ml) ble det ved ca. -ICC under nitrogen satt diisopropyletylamin (0,42 ml, 2,4 mmol) fulgt av difenylklorfosfat (0,50 ml, 2,4 mmol). Blandingen ble omrørt ved 0°C i 30 minutter og derefter avkjølt til To a solution of (5R)-allyl-3,7-dioxo-(6S)-[1-(R)-hydroxyethyl]-1-azabicyclo(3.2.0)heptane-2-(2R)-carboxylate (473 mg , 1.87 mmol) in CH3CN (6 ml) at approx. -ICC under nitrogen added diisopropylethylamine (0.42 mL, 2.4 mmol) followed by diphenylchlorophosphate (0.50 mL, 2.4 mmol). The mixture was stirred at 0°C for 30 minutes and then cooled to
—15°C. Til dette ble det satt en oljeaktig suspensjon av N-(2-merkaptoetyl)-pyridiniumklorid (527 mg, 3,00 mmol) i 1 ml CH3CN inneholdende 5 dråper DMF, fulgt av diisopropyletylamin (0,42 ml, 2,4 mmol). Blandingen ble omrørt ved -15°C i 30 minutter, og derefter fortynnet med 20 ml H20. Denne blanding -15°C. To this was added an oily suspension of N-(2-mercaptoethyl)-pyridinium chloride (527 mg, 3.00 mmol) in 1 mL of CH 3 CN containing 5 drops of DMF, followed by diisopropylethylamine (0.42 mL, 2.4 mmol) . The mixture was stirred at -15°C for 30 minutes, then diluted with 20 mL of H 2 O. This mixture
ble renset direkte på en reversfase-silikagel-kolonne (C^g PrepPÅK, 12 g, Waters Associates), og det ble eluert med H2O (100 ml), 30* CH3CN/H20 (100 ml) og til slutt 40* CH3CN/H20 (100 ml). Passende fraksjoner ble samlet, de organiske oppløsningsmidler fjernet ved hjelp av en vakuumpumpe og lyofilisert for å oppnå 786 mg (1,26 mmol, utbytte 67,3*) av tittelforbindelsen som et brunt pulver. was purified directly on a reverse-phase silica gel column (C^g PrepPÅK, 12 g, Waters Associates) and eluted with H2O (100 mL), 30* CH3CN/H2O (100 mL) and finally 40* CH3CN/ H 2 O (100 ml). Appropriate fractions were collected, the organic solvents removed using a vacuum pump and lyophilized to obtain 786 mg (1.26 mmol, yield 67.3*) of the title compound as a brown powder.
■^E-NMR (DMS0-d6, CFT-20) S: 1,16 (3H, d, J-6 Hz, l'-CH3), 2,6-3,7 (m), 3,75-4,3 (2H, m, 5-H og l'-H), 4,65 (2H, m, -CH2CH2-), 4,87 (2H, t, J-6 Hz, -CH2<N®>), 5-6,2 (3H, m, olefiniske protoner), 6,6-7,4 (m, aromatiske protoner), 8,15 (2H, "t", Jx7 Hz, aromatiske protoner meta til nitrogenet), 8,63 (1H, "t", J27 Hz, aromatisk proton para til nitrogenet) og 9,07 (2H, "d", Js:7 Hz, aromatiske protoner orto til nitrogenet) ppm; ■^E-NMR (DMS0-d6, CFT-20) S: 1.16 (3H, d, J-6 Hz, 1'-CH3), 2.6-3.7 (m), 3.75- 4.3 (2H, m, 5-H and 1'-H), 4.65 (2H, m, -CH2CH2-), 4.87 (2H, t, J-6 Hz, -CH2<N®> ), 5-6.2 (3H, m, olefinic protons), 6.6-7.4 (m, aromatic protons), 8.15 (2H, "t", Jx7 Hz, aromatic protons meta to the nitrogen), 8.63 (1H, "t", J27 Hz, aromatic proton para to the nitrogen) and 9.07 (2H, "d", Js:7 Hz, aromatic protons ortho to the nitrogen) ppm;
IR (film) vmaks: 3400 (OH), 1770 (e-laktam), 1690 (ester), IR (film) vmax: 3400 (OH), 1770 (e-lactam), 1690 (ester),
1625 (pyridinio) 1625 (pyridinium)
B. ( 5R)- 3- r( 2- pvridlnloetvl ) tio1-( 6S)- ri-( R)- hvdroksvetvl' l- 7-okso- l- azablcyklo( 3. 2. 0) hept- 2- en- 2- karboksvlat B. ( 5R)- 3- r( 2- pvridlnloetvl ) thio1-( 6S)- ri-( R )- hvdroxwetvl' 1- 7-oxo- 1- azablcyclo( 3. 2. 0) hept- 2- en- 2- carboxylate
Til en oppløsning av (5R)-allyl-3-[(2-pyridinioetyl)tio]-(6S)-[l-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0 )hept-2-en-2-karboksylat-difenylfosfat (156 mg, 0,25 mmol) i CH3CN To a solution of (5R)-allyl-3-[(2-pyridinioethyl)thio]-(6S)-[1-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0 )hept-2 -ene-2-carboxylate diphenyl phosphate (156 mg, 0.25 mmol) in CH3CN
(2 ml) ble det suksessivt ved ca. 22'C satt en oppløsning av kalium-2-etylheksanoat i EtOAc (0,5 M, 0,6 ml, 0,3 mmol), trifenylfosfln (15 mg, 0,057 mmol) og tetrakistrifenylfosfin-palladium (15 mg, 0,013 mmol). Blandingen ble omrørt ved ca. 22°C under nitrogen i 2 timer. Efter tilsetning av 7 ml vannfri Et20 ble bunnfallet filtrert fra, vasket med 7 ml vannfri Et20 og tørket i vakuum under dannelse av 101 mg brunlig, fast stoff. Dette ble renset ved reversfase-kolonnekromatografi (C^g PrepPAK, 12 g, Waters Associates), hvorved man eluerte med H20. Passende fraksjoner (nemlig fraksjonene 7-12, hver på 20 ml) ble samlet og lyofilisert for å oppnå 53 mg (0,16 mmol, utbytte 64*) av tittelforbindelsen som et gulaktig pulver. Dette materialet var for-urenset med kaliumdifenylfosfat og kalium-2-etylheksanoat. (2 ml) it was successively at approx. At 22°C was added a solution of potassium 2-ethylhexanoate in EtOAc (0.5 M, 0.6 mL, 0.3 mmol), triphenylphosphine (15 mg, 0.057 mmol) and tetrakistriphenylphosphine-palladium (15 mg, 0.013 mmol). . The mixture was stirred at approx. 22°C under nitrogen for 2 hours. After the addition of 7 ml of anhydrous Et 2 O, the precipitate was filtered off, washed with 7 ml of anhydrous Et 2 O and dried in vacuo to form 101 mg of brown solid. This was purified by reverse phase column chromatography (C^g PrepPAK, 12 g, Waters Associates), eluting with H 2 O. Appropriate fractions (namely fractions 7-12, 20 mL each) were pooled and lyophilized to obtain 53 mg (0.16 mmol, yield 64*) of the title compound as a yellowish powder. This material was contaminated with potassium diphenyl phosphate and potassium 2-ethyl hexanoate.
<1->H-NMR (D20, CFT-20) 5: 0,80 (t, J-6,4 Hz, Me fra etyl-heksanoat), 1,21 (3H, d, J-6,3 Hz, 1<*->Me), 2,93 (2H, dd, <J>i_4<=>9 Hz, Jgem=4 Hz• 1-Hs), 3,28 (1H, dd, J6_!-6,2 Hz, J6_5-2,5 Hz, 6-H), 3,42 (2H, t, J= 6 Hz, -CH2S), 3,98 (1H, td, <J>s-i-9 Hz, J5_6-2,5 Hz, 5-H), 4,15 (1H, q, J=6,2 Hz, l'-H), 4,80 (2H, t, J=6,0 Hz, -CH2N<®>), 7-7,5 (m, fenylprotoner fra difenylfosfat), 8,03 (2H, m, Hm av pyridinium), 8,56 (1H, m, Hp av pyridinium) og 8,81 (2H, "d", Jx6,5 Hz, Ho av pyridinium) ppm. <1->H-NMR (D20, CFT-20) δ: 0.80 (t, J-6.4 Hz, Me from ethyl hexanoate), 1.21 (3H, d, J-6.3 Hz , 1<*->Me), 2.93 (2H, dd, <J>i_4<=>9 Hz, Jgem=4 Hz• 1-Hs), 3.28 (1H, dd, J6_!-6, 2 Hz, J6_5-2.5 Hz, 6-H), 3.42 (2H, t, J= 6 Hz, -CH2S), 3.98 (1H, td, <J>s-i-9 Hz, J5_6- 2.5 Hz, 5-H), 4.15 (1H, q, J=6.2 Hz, l'-H), 4.80 (2H, t, J=6.0 Hz, -CH2N<® >), 7-7.5 (m, phenyl protons from diphenyl phosphate), 8.03 (2H, m, Hm of pyridinium), 8.56 (1H, m, Hp of pyridinium) and 8.81 (2H, "d ", Jx6.5 Hz, Ho of pyridinium) ppm.
Eksempel 16 Example 16
Fremstilling av 3- r2-( N- metvl- tiomorfollnium) etvltlo1- 6a- ri'-( R)- hydroksyetyl1- 7- okso- l- azabicyklo( 3. 2. 0) hept- 2- en- 2-karboks<y>lat Preparation of 3-r2-(N-metvl-thiomorpholnium)etvltlo1-6a-ri'-(R)-hydroxyethyl1-7-oxo-1-azabicyclo(3.2.0)hept-2-ene-2-carbox< y>lazy
A. N- metyl- N-( 2- merkaptoetyl)- tlomorfolinlummetansulfonat A. N-Methyl-N-(2-mercaptoethyl)-tlomorpholinelummethanesulfonate
Til Isbad foravkjølt N-metyltiomorfolin<5> (5,00 g, 42,7 mmol) ble det satt metansulfonsyre (1,47 ml, 20,5 mmol) og etylensulfid (1,30 ml, 21,4 mmol). Blandingen ble oppvarmet ved 65°C i 24 timer og fortynnet med 25 ml vann. Den vandige oppløsning ble vasket med 3 x 25 ml dietyleter, pumpet under vakuum og helt over en silikagel-reversfase-kolonne, idet tittelforbindelsen ble eluert med vann. De passende fraksjoner ble forenet og dampet inn under dannelse av tiol som en olje (4,80 g, utbytte 86*). Methanesulfonic acid (1.47 ml, 20.5 mmol) and ethylene sulfide (1.30 ml, 21.4 mmol) were added to ice-bath precooled N-methylthiomorpholine<5> (5.00 g, 42.7 mmol). The mixture was heated at 65°C for 24 hours and diluted with 25 ml of water. The aqueous solution was washed with 3 x 25 mL of diethyl ether, pumped under vacuum and poured over a silica gel reverse phase column, eluting the title compound with water. The appropriate fractions were combined and evaporated to give the thiol as an oil (4.80 g, yield 86*).
IR (film) vmaks: 2550 (w, SH) cm-1;IR (film) vmax: 2550 (w, SH) cm-1;
1-H-NMR (DMS0-d6) S: 3,25-2,95 (6H, m, CH2N<®>), 3,20-2,65 (7H, 1-H-NMR (DMS0-d6) S: 3.25-2.95 (6H, m, CH2N<®>), 3.20-2.65 (7H,
m, CH2S, SH) og 2,32 (3H, s, CH3SO3) ppm. m, CH2S, SH) and 2.32 (3H, s, CH3SO3) ppm.
<5> J.M. Lehn & J. Wagner, "Tetrahedron", 26, 4227 <5> J.M. Lehn & J. Wagner, "Tetrahedron", 26, 4227
(1970). (1970).
B. p- nitrobenzyl- 3- r2-( N- metyltiomorfolinlum- dlfenvlfosfat) etvltio1- 6a- ri'-( R)- hvdroksvetvl1- 7- okso- l-azabicyklo( 3. 2. 0) hept- 2- en- 2- karboksvlat B. p- nitrobenzyl- 3- r2-( N- methylthiomorpholinelum- dlphenvlphosphate) ethvlthio1- 6a- ri'-( R)- hvdroxvevl1- 7- oxo- l-azabicyclo( 3. 2. 0) hept- 2- en- 2- carboxylate
En kald (isbad) oppløsning av p-nitrobenzyl-6a-[1'-(R)-hydroksyetyl]-3,7-diokso-l-azabicyklo(3.2.0)heptan-2-karboksylat (557 mg, 1,60 mmol) 1 8 ml CH3CN ble dråpevis behandlet med diisopropyletylamin (0,336 ml, 1,92 mmol) og difenylklorfosfat (0,400 ml, 1,92 mmol) og omrørt i 30 minutter. Reaksjonsblandingen ble på ny behandlet med N-metyl-N-(2-merkaptoetyl)tiomorfolinium-metansulfonat (893 mg, 2,29 mmol) i 4 ml CH3CN og diisopropyletylamin (0,336 ml, 1,92 mmol) og omrørt i 30 minutter. Oppløsningen ble fortynnet med 20 ml vann og helt over en silikagel-reversfase-kolonne. Den ønskede forbindelse ble fortynnet med en 50* acetonitril-vann-blanding. De passende fraksjoner ble forenet, pumpet under vakuum i 2 timer og lyofilisert under dannelse av tittelforbindelsen (1,01 g, utbytte 85*). A cold (ice bath) solution of p-nitrobenzyl-6α-[1'-(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-carboxylate (557 mg, 1.60 mmol) 18 mL of CH3CN was treated dropwise with diisopropylethylamine (0.336 mL, 1.92 mmol) and diphenylchlorophosphate (0.400 mL, 1.92 mmol) and stirred for 30 minutes. The reaction mixture was again treated with N-methyl-N-(2-mercaptoethyl)thiomorpholinium methanesulfonate (893 mg, 2.29 mmol) in 4 mL of CH 3 CN and diisopropylethylamine (0.336 mL, 1.92 mmol) and stirred for 30 minutes. The solution was diluted with 20 ml of water and poured over a silica gel reverse phase column. The desired compound was diluted with a 50% acetonitrile-water mixture. The appropriate fractions were combined, pumped under vacuum for 2 hours and lyophilized to give the title compound (1.01 g, yield 85*).
IR (nujol) <v>maks: 1760 (s, e-laktam C-0) og 1510 (s, N02) IR (nujol) <v>max: 1760 (s, e-lactam C-0) and 1510 (s, N02)
cm<-1>; <*>H-NMR (DMS0-d6) S: 8,25 (2H, d, J-8,8 Hz, H-aromatisk), 7,70 (2H, d, J-8,8 Hz, H-aromatisk), 7,33-6,84 (10H, m, H-aromatisk), 5,37 (2H, sentrum for ABq, J-14,2 Hz, CH2), 5,14 (1H, d, J-4,5 Hz, OH), 4,35-3,80 (2H, m, H-l' og H-5), 3,75-3,45 (6H, m, CH2N<®>), 3,31 (3H, s, CH3N<®>), 3,45-2,75 (9H, m, CH2S, H-6 og H-4) og 1,15 (3H, d, J-6,2 Hz, CH3) ppm. C. 3- r2-( N- metvl- tiomorfolinlum) etvltlo1- 6a- ri,-( R)- hvdroksv etvll- 7- okso- l- azabicyklo( 3. 2. 0) hept- 2- en- 2- karboksylat cm<-1>; <*>H-NMR (DMS0-d6) S: 8.25 (2H, d, J-8.8 Hz, H-aromatic), 7.70 (2H, d, J-8.8 Hz, H- aromatic), 7.33-6.84 (10H, m, H-aromatic), 5.37 (2H, center of ABq, J-14.2 Hz, CH2), 5.14 (1H, d, J- 4.5 Hz, OH), 4.35-3.80 (2H, m, H-1' and H-5), 3.75-3.45 (6H, m, CH2N<®>), 3.31 ( 3H, s, CH3N<®>), 3.45-2.75 (9H, m, CH2S, H-6 and H-4) and 1.15 (3H, d, J-6.2 Hz, CH3) ppm. C. 3- r2-( N- metvl- thiomorpholinum) etvltlo1- 6a- ri,-( R)- hvdroksv etvll- 7- oxo- l- azabicyclo( 3. 2. 0) hept- 2- en- 2- carboxylate
En oppløsning av p-nitrobenzyl-3-[2-(N-metyl-tiomorfolinium-difenylfosfat)etyltio]-6a-[l'-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat (1,31 g, 1,76 mmol) i 0,1 M fosfatbuffer, pH 7,4 (48,8 ml), tetrahydrofuran (20 ml) og dietyleter (20 ml) ble hydrogenert over 1,5 g 10* palladium-på-trekull i et Parr-rysteapparat i 1 time ved ca. 2,8 kp/cm<2>. Reaksjonsblandingen ble fortynnet med 40 ml dietyleter og fasene separert. Den organiske fase ble vasket med 2 x 5 ml vann. De vandige faser ble kombinert, filtrert gjennom filterpapir som var herdet 52 ganger, vasket med 2 x 50 ml dietyleter og pumpet under vakuum. Den vandige oppløsning ble helt på en silikagel-reversfase-kolonne og det ønskede karbapenem ble fortynnet med 5* acetonitril-vann. De passende fraksjoner ble kombinert og lyofilisert og man oppnådde tittelforbindelsen som et amorft, fast stoff (205 mg, 31*). A solution of p-nitrobenzyl-3-[2-(N-methyl-thiomorpholinium-diphenylphosphate)ethylthio]-6a-[1'-(R)-hydroxyethyl]-7-oxo-1-azabicyclo(3.2.0)hept -2-ene-2-carboxylate (1.31 g, 1.76 mmol) in 0.1 M phosphate buffer, pH 7.4 (48.8 mL), tetrahydrofuran (20 mL) and diethyl ether (20 mL) was hydrogenated over 1.5 g 10* palladium-on-charcoal in a Parr shaker for 1 hour at approx. 2.8 kp/cm<2>. The reaction mixture was diluted with 40 ml of diethyl ether and the phases separated. The organic phase was washed with 2 x 5 ml of water. The aqueous phases were combined, filtered through filter paper hardened 52 times, washed with 2 x 50 ml of diethyl ether and pumped under vacuum. The aqueous solution was poured onto a silica gel reverse phase column and the desired carbapenem was diluted with 5* acetonitrile-water. The appropriate fractions were combined and lyophilized to give the title compound as an amorphous solid (205 mg, 31*).
IR (nujol) vmaks: 1750 (s, e-laktam C-0) og 1590 (s, C-0) cm<-1>; IR (nujol) vmax: 1750 (s, ε-lactam C-0) and 1590 (s, C-0) cm<-1>;
<i>H-NMR (D20) S: 4,25-3,95 (2H, m, H-l' , H-5), 3,70-3,40 (6H, <i>H-NMR (D 2 O) S: 4.25-3.95 (2H, m, H-1' , H-5), 3.70-3.40 (6H,
m, CH2N<®>), 3,35 (1H, dd, J-6,1 Hz, J-2,6 Hz, H-6), 3,08 (3H, s, CH3N<®>), 3,25-2,75 (8H, CH2S, H-4) og 1,24 ppm (3H, d, J-6,4 Hz, CH3); m, CH2N<®>), 3.35 (1H, dd, J-6.1 Hz, J-2.6 Hz, H-6), 3.08 (3H, s, CH3N<®>), 3 .25-2.75 (8H, CH2S, H-4) and 1.24 ppm (3H, d, J-6.4 Hz, CH3);
UV (H20, c 0,062) Xmaks: 299 (cl0962) nm; UV (H 2 O, c 0.062) Xmax: 299 (cl 0962) nm;
t% - 17,7 timer (0,1 M fosfatbuffer, pH 7, 37'C). t% - 17.7 hours (0.1 M phosphate buffer, pH 7, 37°C).
Eksempel 17 Example 17
Fremstilling av ( 5R. 6S)- 3- T2-( 1- metvlmorfollno) etvltiol- 6-f( R)- l- hvdroksvetvl1- 7- okso- l- azabicvklo( 3. 2. 0) hent- 2- en- 2-karboksylat Preparation of (5R.6S)-3-T2-(1-methylmorpholno)ethylthiol-6-f(R)-1-hydroxymethyl-7-oxo-1-azabiclo(3.2.0)hent-2-en- 2-carboxylate
A. l- metvl- l-( 2- merkaptoetvl) morfollnlum- trifluormetan-sulfonat A. 1- Methyl- 1-(2-mercaptoethyl) morpholine trifluoromethanesulfonate
Til N-metylmorfolin (3,29 ml, 0,030 mol) ble det dråpevis satt trifluormetansulfonsyre (1,327 ml, 0,015 mol) ved 10"C, fulgt av etylensulfid (0,89 ml, 0,015 mol). Den resulterende gulbrune oppløsning ble oppvarmet i oljebad ved 50-60<*>C under N2 i 18 timer. Flyktige materialer ble derefter fjernet under vakuum, og den resterende olje ble samlet i 10 ml H20. Den vandige oppløsning ble vasket med 3 x 5 ml dietyleter og derefter ble resterende organisk oppløsningsmiddel fjernet under vakuum. Den resulterende vandige oppløsning ble satt til en Cig-reversfase-kolonne, det ble fortynnet med H20 og derefter med 5* acetonitril-H20 og til slutt med 10* acetonltril-H20. Avdamping av de relevante fraksjoner ga et hvitt, fast stoff som ble tørket under vakuum over P2C<5, noe som ga produktet (1,92 g, 41*). To N-methylmorpholine (3.29 mL, 0.030 mol) was added dropwise trifluoromethanesulfonic acid (1.327 mL, 0.015 mol) at 10°C, followed by ethylene sulfide (0.89 mL, 0.015 mol). The resulting tan solution was heated in oil bath at 50-60<*>C under N 2 for 18 h. Volatiles were then removed under vacuum and the remaining oil was collected in 10 mL H 2 O. The aqueous solution was washed with 3 x 5 mL diethyl ether and then residual organic solvent removed under vacuum. The resulting aqueous solution was added to a C 10 reverse phase column, it was diluted with H 2 O, then with 5* acetonitrile-H 2 O and finally with 10* acetonitrile-H 2 O. Evaporation of the relevant fractions gave a white , solid which was dried under vacuum over P2C<5 to give the product (1.92 g, 41*).
IR (KBr) vmaks: 2560 (-SH) cm-<1>; IR (KBr) vmax: 2560 (-SH) cm-<1>;
<i>H-NMR (d6-aceton) S: 4,25-3,6 (m, 8H), 3,49 (s, 3H, N-Me), <i>H-NMR (d6-acetone) S: 4.25-3.6 (m, 8H), 3.49 (s, 3H, N-Me),
3,35-2,7 (m, 5H). 3.35-2.7 (m, 5H).
B. p- nltrobenzvl-( 5R. 6S)- 3- f2-( 1- metvlmorfolino) etvltlol- 6-f( R)- l- hvdroksvetyl1- 7- okso- l- azabicyklo( 3. 2. 0) hept- 2- en-2- karboksvlat- difenvlfosfat B. p- niltrobenzvl-( 5R. 6S)- 3- f2-( 1- methylmorpholino) ethyllol- 6-f( R)- l- hydroxybutyl1- 7- oxo- l- azabicyclo( 3. 2. 0) hept- 2- en-2- carboxyvlate- diphenyl phosphate
Til en oppløsning av p-nitrobenzyl-(5R,6S)-6-[(R)-l-hydroksy-etyl]-3,7-diokso-l-azabicyklo(3.2.0)heptan-2-karboksylat (0,348 g, 1,0 mmol) i 25 ml tørr acetonitril ble det dråpevis satt diisopropyletylamin (0,191 ml, 1,1 mmol) ved 0<*>C under N2. Efter omrøring ved 0"C i 1 time ble det tilsatt diisopropyletylamin (0,226 ml, 1,3 mmol) til det resulterende enol-fosfat, fulgt av l-metyl-l-(2-merkaptoetyl)morfolinium-trifluormetansulfonat (0,373 g, 1,2 mmol). Reaksjonsblandingen ble omrørt ved romtemperatur i 1,5 timer og derefter konsentrert under vakuum. Det resterende materialet ble samlet i H20 og satt til en C^g-reversfase-kolonne. For-tynning med H20 og derefter med 20* acetonitril-H20 og til slutt med 30* acetonitril-H20, fulgt av lyofilisering av de relevante fraksjoner ga produktet (0,360 g, 40*) som et amorft, fast stoff. To a solution of p-nitrobenzyl-(5R,6S)-6-[(R)-1-hydroxy-ethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2-carboxylate (0.348 g , 1.0 mmol) in 25 ml of dry acetonitrile, diisopropylethylamine (0.191 ml, 1.1 mmol) was added dropwise at 0<*>C under N2. After stirring at 0°C for 1 hour, diisopropylethylamine (0.226 mL, 1.3 mmol) was added to the resulting enol phosphate, followed by 1-methyl-1-(2-mercaptoethyl)morpholinium trifluoromethanesulfonate (0.373 g, 1 .2 mmol). The reaction mixture was stirred at room temperature for 1.5 hours and then concentrated under vacuum. The remaining material was taken up in H 2 O and applied to a C 2 g reverse phase column. Diluting with H 2 O and then with 20* acetonitrile-H 2 O and finally with 30* acetonitrile-H 2 O, followed by lyophilization of the relevant fractions gave the product (0.360 g, 40*) as an amorphous solid.
IR (film) vmaks: 3300 (-0H), 1770 (e-laktam CO), 1700 IR (film) vmax: 3300 (-0H), 1770 (e-lactam CO), 1700
(-C02pNB) cm-<1>;(-CO 2 pNB) cm-<1>;
^H-NMR (d6-aceton) S: 8,25, 7,80 (ABq, J-8,6 Hz, 4H, ^H-NMR (d6-acetone) S: 8.25, 7.80 (ABq, J-8.6 Hz, 4H,
aromatisk), 7,4-6,8 (m, 10H, difenylfosfat), 5,56, 5,27 (ABq, J-14,2 Hz, 2H, benzylisk), 4,42 (d av t, J-9,2 Hz, J'-2,7 Hz, 1H, H-5), 4,1-2,7 (m, 17H), 3,40 (s, 3H, N-Me), 1,22 (d, J-6,2 Hz, 3H, -CHMe). aromatic), 7.4-6.8 (m, 10H, diphenyl phosphate), 5.56, 5.27 (ABq, J-14.2 Hz, 2H, benzylic), 4.42 (d of t, J- 9.2 Hz, J'-2.7 Hz, 1H, H-5), 4.1-2.7 (m, 17H), 3.40 (s, 3H, N-Me), 1.22 ( d, J-6.2 Hz, 3H, -CHMe).
C. ( 5R. 6S)- 3- f2-( 1- metvlmorfolino) etvltio1- 6- f( R)- l- hvdroksvetvll- 7- okso- l- azabicvklo( 3. 2. 0) hept- 2- en- 2- karboksvlat C. ( 5R. 6S)- 3- f2-( 1- methylmorpholino) ethylthiol- 6- f( R)- 1- hydroxymethyl- 7- oxo- 1- azabicchloro( 3. 2. 0) hept- 2- en- 2- carboxylate
Til en oppløsning av p-nitrobenzyl-(5R,6S)-3-[2-(l-metyl-morfolino)etyltio]-6-[(R)-l-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat-difenylfosfat (0,360 g, 0,49 mmol) i 13 ml fosfatbuffer (0,05 M, pH 7,4) ble det satt 0,36 g 10* palladium-på-trekull, 20 ml tetrahydrofuran og 20 ml dietyleter. Denne blanding ble hydrogenert i en Parr-apparatur ved 2,24 kp/cm<2> i 1 time. Blandingen ble filtrert gjennom Celite, og filterputen vasket med H2O og dietyleter. Den vandige fase ble separert og pH-verdien innstilt til 7,0 med ytterligere fosfatbuffer, pH 7,4. Efter fjerning av tilbakeblevne organiske oppløsningsmidler under vakuum, ble den vandige oppløsning satt til en Cig-reversfase-kolonne. Eluering med H20 og lyofilisering av de relevante fraksjoner ga 0,13 f av et amorft, fast stoff. Dette ble renset på ny ved reversfase-HPLC, noe som ga det rene produkt (0,058 g, 34*) som et amorft, fast stoff. To a solution of p-nitrobenzyl-(5R,6S)-3-[2-(1-methyl-morpholino)ethylthio]-6-[(R)-1-hydroxyethyl]-7-oxo-1-azabicyclo(3.2 .0)hept-2-ene-2-carboxylate-diphenyl phosphate (0.360 g, 0.49 mmol) in 13 ml of phosphate buffer (0.05 M, pH 7.4) was added to 0.36 g of 10* palladium -charcoal, 20 ml of tetrahydrofuran and 20 ml of diethyl ether. This mixture was hydrogenated in a Parr apparatus at 2.24 kp/cm<2> for 1 hour. The mixture was filtered through Celite, and the filter pad washed with H 2 O and diethyl ether. The aqueous phase was separated and the pH adjusted to 7.0 with additional phosphate buffer, pH 7.4. After removal of residual organic solvents under vacuum, the aqueous solution was applied to a Cig reverse phase column. Elution with H 2 O and lyophilization of the relevant fractions gave 0.13 g of an amorphous solid. This was purified again by reverse phase HPLC to give the pure product (0.058 g, 34*) as an amorphous solid.
IR (KBr) vmaks: 3420 (br, OH), 1750 (e-laktam CO), 1590 IR (KBr) vmax: 3420 (br, OH), 1750 (e-lactam CO), 1590
(-C02<e>) cm"<1>; (-C02<e>) cm"<1>;
<i>H-NMR (D20) S: 4,35-2,77 (m, 17H), 3,18 (s, 3H, N-Me), <i>H-NMR (D 2 O) S: 4.35-2.77 (m, 17H), 3.18 (s, 3H, N-Me),
1,23 (d, J=6,3 Hz, 3H, CHMe); 1.23 (d, J=6.3 Hz, 3H, CHMe);
UV (H20) Xmaks: 300 (c6344) nm; UV (H2O) Xmax: 300 (c6344) nm;
ty, (pH 7,4, 36,8'C), 18,5 t. ty, (pH 7.4, 36.8°C), 18.5 h.
Eksempel 18 Example 18
Fremstilling av ( 5R. 6S)- 3- T2-( 1. 4- dimetyl- l- plperazinlum)-etvltlo1- 6- ri-( R)- hvdroksvetvl1- 7- okso- l- azabicvklof 3. 2. 0)-hept- 2- en- 2- karboksylat Preparation of (5R.6S)-3-T2-(1.4-Dimethyl-1-plperazine)-ethvltlo1-6-ri-(R)-hydroxvetl1-7-oxo-1-azabicvklof 3.2.0)- hept-2-ene-2-carboxylate
A. l-( 2- acetyltioetyl )- l . 4- dimetylplperaziniumbromid A. l-(2-Acetylthioethyl)-l. 4- dimethylplperazinium bromide
En oppløsning av 2-brometyltioacetat^ (2,20 g, 0,012 mol) og 1,4-dimetylpiperazin (1,95 ml, 0,014 mol) i 4 ml aceton ble omrørt ved 50<*>C i 65 timer. Efter avkjøling til 25°C ble den flytende fase omhelt fra gummien som var triturert to ganger i dietyleter, og man oppnådde et hygroskopisk, gulaktig pulver, 3,2 g (90*). A solution of 2-bromomethylthioacetate^ (2.20 g, 0.012 mol) and 1,4-dimethylpiperazine (1.95 mL, 0.014 mol) in 4 mL of acetone was stirred at 50<*>C for 65 hours. After cooling to 25°C, the liquid phase was decanted from the gum which had been triturated twice in diethyl ether, and a hygroscopic yellowish powder was obtained, 3.2 g (90*).
IR (nujol) vmaijS: 1685 (C=0 av tloester) cm-<1>; IR (nujol) vmaijS: 1685 (C=0 of tloester) cm-<1>;
0 0
■^H-NMR (D20) S: 2,37, 2,39 (2s, 6H, CH3CO, ■^H-NMR (D 2 O) S: 2.37, 2.39 (2s, 6H, CH 3 CO,
<6> B. Hansen, "Acta Chem. Scand.", 11, 537-40 (1957). <6> B. Hansen, "Acta Chem. Scand.", 11, 537-40 (1957).
B. 1. 4- dlmetvl- l-( 2- merkaptoetvl) piperaziniumbromid. B. 1. 4-Dimethyl-1-(2-mercaptoethyl)piperazinium bromide.
hvdroklorld hydrogen chloride
En oppløsning av l-(2-acetyltioetyl)-l,4-dimetyl-piperaziniumbromid (1,1 g, 3,7 mmol) i 6N saltsyre (4 ml) ble oppvarmet til 80<4>C under nitrogen i 1 time. Oppløsningen ble konsentrert under redusert trykk, noe som ga et hvitt pulver, 0,41 g (38*); <1->H-NMR (DMS0-d6) S: 2,90 (s 3,26 (s, A solution of 1-(2-acetylthioethyl)-1,4-dimethyl-piperazinium bromide (1.1 g, 3.7 mmol) in 6N hydrochloric acid (4 mL) was heated to 80<4>C under nitrogen for 1 hour. The solution was concentrated under reduced pressure to give a white powder, 0.41 g (38*); <1->H-NMR (DMS0-d6) S: 2.90 (s 3.26 (s,
Analyse for CgB^o^SBrCl 'B^O: Analysis for CgB^o^SBrCl 'B^O:
Beregnet: C 31,03 H 7,16 N 9,05 S 10,35 Funnet: C 31,62 H 7,46 N 9,19 S 10,19 C. ( 5R. 6S)- p- nitrobenzvl- 3- r2-( 1. 4- dlmetvl- l- plperazinlum)-etvltiol- 6- fl-( R)- hvdroksyetvl1- 7- okso- l- azablcyklo-( 3. 2. 0) hept- 2- en- 2- karboksvlat- difenylfosfat Calculated: C 31.03 H 7.16 N 9.05 S 10.35 Found: C 31.62 H 7.46 N 9.19 S 10.19 C. ( 5R. 6S)- p- nitrobenzvl- 3- r2-(1.4-dlmethyl-1-plperazine)-ethylthiol-6-fl-(R)-hydroxyethyl1-7-oxo-1-azablcyclo-(3.2.0)hept-2-ene-2-carboxylate - diphenyl phosphate
Til en kald (0<*>C) oppløsning av (5R,6S)-p-nitrobenzyl-6-[l-(R)-hydroksyetyl]-3,7-diokso-l-azabicyklo(3.2.0)heptan-2-(R)-karboksylat (0,465 g, 1,33 mmol) 1 2 ml acetonitril, holdt under nitrogen, ble det tilsatt diisopropyletylamin (0,278 ml, 1,59 mmol) og difenylklorfosfat (0,33 ml, 1,59 mmol). Reaksjonsblandingen ble omrørt 1 30 minutter og behandlet med en suspensjon av 1,4-dimetyl-l-(2-merkaptoetyl)piperazinlum-bromid.hydroklorid (0,40 g, 1,37 mmol) 1 en blanding av 3 ml acetonitril og 1 ml vann og diisopropyletylamin (0,278 ml, 1,59 mmol). Efter omrøring i 18 timer ved 5'C satte man 15 ml kaldt vann til blandingen. Den resulterende oppløsning ble kromatografert over en PrepPAK -500/Cig-kolonne (Waters Associates) (2,5 x 7,5 cm) med 25-35* acetonitril i vann som elueringsmiddel, noe som ga et gulaktig pulver, 0,50 g (50*) efter lyofilisering. To a cold (0<*>C) solution of (5R,6S)-p-nitrobenzyl-6-[l-(R)-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane- 2-(R)-Carboxylate (0.465 g, 1.33 mmol) 1 To 2 mL of acetonitrile, kept under nitrogen, was added diisopropylethylamine (0.278 mL, 1.59 mmol) and diphenyl chlorophosphate (0.33 mL, 1.59 mmol ). The reaction mixture was stirred for 130 minutes and treated with a suspension of 1,4-dimethyl-1-(2-mercaptoethyl)piperazinum bromide hydrochloride (0.40 g, 1.37 mmol) 1 a mixture of 3 mL of acetonitrile and 1 ml of water and diisopropylethylamine (0.278 ml, 1.59 mmol). After stirring for 18 hours at 5°C, 15 ml of cold water was added to the mixture. The resulting solution was chromatographed over a PrepPAK -500/Cig column (Waters Associates) (2.5 x 7.5 cm) eluting with 25-35% acetonitrile in water to give a yellowish powder, 0.50 g (50*) after lyophilization.
IR (KBr) vmaks: 1765 (C-0 av e-laktam), 1690 (C-0 av pNB-ester), 1585 (fenyl), 1512 (N02), 875 (N02) cm-<1>; IR (KBr) vmax: 1765 (C-0 of ε-lactam), 1690 (C-0 of pNB ester), 1585 (phenyl), 1512 (N02), 875 (N02) cm-<1>;
^H-NMR (DMS0-d6) 5: 1,16, 1,18 (2d, J-6,1 Hz, 3H, CH3CH0H), ^H-NMR (DMSO-d6) δ: 1.16, 1.18 (2d, J-6.1 Hz, 3H, CH3CH0H),
2,44 (s 2.44 (p
3,14 (s, 3.14 (s,
5,31 (d, 5.31 (d,
J-6 Hz, OH), 5,39 (sentrum for ABq, J-13 Hz, CH2 av pNB), 6,6-7,4 (m, 10H, fenyl av fosfat), 7,71 (d, J-8,8 Hz, 2H, Ho av pNB), 8,26 (d, J-8,8 Hz, Hm av pNB). J-6 Hz, OH), 5.39 (center of ABq, J-13 Hz, CH2 of pNB), 6.6-7.4 (m, 10H, phenyl of phosphate), 7.71 (d, J -8.8 Hz, 2H, Ho of pNB), 8.26 (d, J-8.8 Hz, Hm of pNB).
D. ( 5R. 6Sl- 3- T2-( 1. 4- dimetvl- l- plperazlniiup) etvltlo1- 6- ri-( R)- hydroksyetyl1- 7- okso- l- azablcvklo( 3. 2. 0) hept- 2- en- 2-karboksylat D. ( 5R. 6Sl- 3- T2-( 1. 4- dimetvl- l- plperazlniiup) etvltlo1- 6- ri-( R)- hydroxyethyl1- 7- oxo- l- azablcvclo( 3. 2. 0) hept- 2- en- 2-carboxylate
Til en oppløsning av (5R,6S)-p-nitrobenzyl-3-[2-(l,4-dimetyl-l-piperazinium)etyltio]-6-[l-(R)-hydroksyetyl]-7-okso-l-azablcyklo(3.2.0)hept-2-en-2-karboksylat-dlfenylfosfat (0,47 g, 0,623 mmol) i 25 ml våt tetrahydrofuran ble det satt 25 ml dietyleter, kaliumfosfat monobasisk-natriumhydroksydbuffer (13 ml, pH 7,22)og 0,47 g 10* palladium-på-trekull. Den resulterende blanding ble hydrogenert ved 23<*>C under ca. 2,8 kp/cm<2> 1 1 time. De to sjikt ble separert og det organiske sjikt ble ekstrahert med 2 x 7 ml vann. De vandige sjikt ble kombinert, filtrert gjennom en Celite-pute, vasket med 2 x 15 ml dietyleter og kromatografert på en PrepPAK -500/Cig-kolonne (Waters Associates) (2,5 x 9,5 cm) med vann som elueringsmiddel, noe som ga 0,097 g (43*) efter lyofilisering. To a solution of (5R,6S)-p-nitrobenzyl-3-[2-(1,4-dimethyl-1-piperazinium)ethylthio]-6-[1-(R)-hydroxyethyl]-7-oxo-1 -azablcyclo(3.2.0)hept-2-ene-2-carboxylate-dlphenylphosphate (0.47 g, 0.623 mmol) in 25 ml of wet tetrahydrofuran was added 25 ml of diethyl ether, potassium phosphate monobasic sodium hydroxide buffer (13 ml, pH 7, 22) and 0.47 g of 10* palladium-on-charcoal. The resulting mixture was hydrogenated at 23<*>C under approx. 2.8 kp/cm<2> 1 1 hour. The two layers were separated and the organic layer was extracted with 2 x 7 ml of water. The aqueous layers were combined, filtered through a Celite pad, washed with 2 x 15 mL of diethyl ether, and chromatographed on a PrepPAK -500/Cig column (Waters Associates) (2.5 x 9.5 cm) with water as eluent, which gave 0.097 g (43*) after lyophilization.
IR (KBr) vmaks: 3000-3700 (OH), 1750 (C-0 av e-laktam), IR (KBr) vmax: 3000-3700 (OH), 1750 (C-0 of ε-lactam),
1585 (karboksylat) cm-<1>; 1585 (carboxylate) cm-<1>;
^H-NMR (D20) S: 1,24 (d, J-6,4 Hz, 3H, CHoCHOH), 2,33 (s, ^H-NMR (D 2 O) S: 1.24 (d, J-6.4 Hz, 3H, CHoCHOH), 2.33 (s,
3,15 (s, 3.15 (s,
4,0-4,5 (m, 4.0-4.5 (m,
UV (H20) Xmaks: 296 (c9476) nm; UV (H 2 O) Xmax: 296 (c9476) nm;
[a]<g3> 61,1° (c 0,26, H20); [α]<g3> 61.1° (c 0.26, H 2 O);
t^ =• 12,4 timer (målt ved en konsentrasjon på 10~<4> M 1 t^ =• 12.4 hours (measured at a concentration of 10~<4> M 1
fosfatbuffer, pH 7,4, ved 36,8<*>C). phosphate buffer, pH 7.4, at 36.8<*>C).
Eksempel 19 Example 19
Fremstilling av ( 5R. 6S)- 3- r2-( N- metyl- tiomorfolinlumoksydl-etvltiol- 6- fl-( R)- hydroksyetyl1- 7- okso- l- azabicvklo( 3. 2. 0 )-hept- 2- en- 2- karboksvlat Preparation of (5R.6S)-3-r2-(N-methyl-thiomorpholinelumoxydl-ethvlthiol-6-fl-(R)-hydroxyethyl1-7-oxo-l-azabiclo(3.2.0)-hept-2- a- 2- carboxylate
Til en kald (-10'C) oppløsning av (5R,6S)-3-[2-(N-metyltio-morfollnium)etyltio]-6-[l-(R)-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylat (6,08 mg, 1,65 mmol) i 9 ml av en l:l-blanding av acetonitril og vann ble det satt m-klorperbenzosyre (334,8 mg, 1,65 mmol) i liten mengde i løpet av 1 time. Blandingen ble derefter fortynnet med 15 ml vann og vasket med 3 x 15 ml dietyleter. Den vandige fase ble pumpet under vakuum og ført gjennom en reversfase-silikagel-kolonne (H2O), noe som ga et fast stoff som bestod av en blanding av forbindelser. Denne blanding ble separert ved reversfase-HPLC, noe som ga fraksjon A, 52,4 mg (utbytte 12*) og fraksjon B, 23,6 mg (utbytte 6*) som diastereoisomerer av tittelforbindelsen; To a cold (-10'C) solution of (5R,6S)-3-[2-(N-methylthio-morpholnium)ethylthio]-6-[1-(R)-hydroxyethyl]-7-oxo-1- azabicyclo(3.2.0)hept-2-ene-2-carboxylate (6.08 mg, 1.65 mmol) in 9 ml of a 1:1 mixture of acetonitrile and water was added m-chloroperbenzoic acid (334.8 mg, 1.65 mmol) in a small amount during 1 hour. The mixture was then diluted with 15 ml of water and washed with 3 x 15 ml of diethyl ether. The aqueous phase was pumped under vacuum and passed through a reverse phase silica gel (H 2 O) column, yielding a solid consisting of a mixture of compounds. This mixture was separated by reverse phase HPLC to give fraction A, 52.4 mg (yield 12*) and fraction B, 23.6 mg (yield 6*) as diastereoisomers of the title compound;
fraksjon A: fraction A:
IR (nujol) vmaks: 1750 (s, e-laktam C-0) og 1580 (s, C-0) cm-1;IR (nujol) vmax: 1750 (s, ε-lactam C-0) and 1580 (s, C-0) cm-1;
%-NMR (D20) 5: 4,26-2,91 (20H, m, H-4, H-5, H-6, E- I', CH2S, %-NMR (D2O) 5 : 4.26-2.91 (20H, m, H-4, H-5, H-6, E- I', CH2S,
CH2S-0, CH3-N<®> og CH2-N<®>) og 1,24 (3H, d, J-6,4 Hz, CH3) ppm; CH2S-O, CH3-N<®> and CH2-N<®>) and 1.24 (3H, d, J-6.4 Hz, CH3) ppm;
UV (H20, c 0,06) Xmaks: ^02 (cl0425) nm; UV (H2O, c 0.06) Xmax: ^02 (cl0425) nm;
t^: 12 timer (0,065 M, pH 7,4, fosfatbuffer, 37°C). t^: 12 hours (0.065 M, pH 7.4, phosphate buffer, 37°C).
fraksjon B: faction B:
IR (nujol) <v>maks: 1750 (s, e-laktam, C-0) og 1585 (s, C-0) cm-*;IR (nujol) <v>max: 1750 (s, ε-lactam, C-0) and 1585 (s, C-0) cm-*;
<1->H-NMR (D20) S: 3,86-2,90 (17H, m, H-4, H-5, H-6, H-l' , CH2S, <1->H-NMR (D2O) S: 3.86-2.90 (17H, m, H-4, H-5, H-6, H-1' , CH2S,
CH2S-0, CH2N<®>), 3,25 (3H, s, CH3N<®>) og 1,24 (3H, d, J=6,4 Hz, CH3) p<p>m; CH2S-0, CH2N<®>), 3.25 (3H, s, CH3N<®>) and 1.24 (3H, d, J=6.4 Hz, CH3) p<p>m;
UV (H20, c 0,05) Xmaks: 2,99 (c6517) nm; UV (H 2 O, c 0.05) Xmax: 2.99 (c6517) nm;
t^: 10,75 timer (0,065 M, pH 7,4, bufferoppløsning, 37°C). t^: 10.75 hours (0.065 M, pH 7.4, buffer solution, 37°C).
Eksempel 20 Example 20
Fremstilling av ( 5R. 6S)- 3- T2-( l. 4. 4- trlmetyl- l- pipera-zinlum) etyltio" l- 6-[" lR- hydroksyetyl1 - 7- okso- l- azabicyklo-( 3. 2. 0) hept- 2- en- 2- karboksvlatklorid Preparation of (5R.6S)-3-T2-(1.4.4-trimethyl-1-piperazinelum)ethylthio"1-6-["1R-hydroxyethyl1-7-oxo-1-azabicyclo-(3. 2. 0) hept- 2- en- 2- carboxyvlate chloride
A. l-( 2- acetyltloetvl )- l . 4 . 4- trimetylplperazinium- bromid. 1odld A. l-(2-Acetyltloethyl)-l. 4. 4-trimethylplperazinium bromide. 1
En suspensjon av l-(2-acetyltioetyl)-l,4-dimetyl-pipera-zlnlumbromld (1,48 g, 5,0 mmol) i 10 ml isopropylalkohol ble behandlet med metyljodld (0,373 ml, 6,0 mmol) og oppvarmet til 55-60°C 1 30 timer. Oppløsningsmidlene ble dampet av under redusert trykk, resten trlturert 1 heksan og det faste stoff filtrert, man oppnådde 1,85 g. Det faste stoff ble oppløst i 8 ml varmt vann og oppløsningen fortynnet med aceton til uklarhet (70-80 ml). To efterfølgende krystalli-seringer ga 1,5 g med smeltepunkt 220-225"C under dekompo-nering, 68* av tittelforbindelsen. A suspension of 1-(2-acetylthioethyl)-1,4-dimethyl-pipera-zlnlumbromide (1.48 g, 5.0 mmol) in 10 mL of isopropyl alcohol was treated with methyl iodide (0.373 mL, 6.0 mmol) and heated to 55-60°C 1 30 hours. The solvents were evaporated under reduced pressure, the residue triturated with 1 hexane and the solid filtered, yielding 1.85 g. The solid was dissolved in 8 ml of hot water and the solution diluted with acetone to cloudiness (70-80 ml). Two subsequent crystallizations gave 1.5 g of m.p. 220-225°C with decomposition, 68% of the title compound.
IR (KBr) vmaks: 1692 (C=0) cm"<1>; IR (KBr) vmax: 1692 (C=0) cm"<1>;
1-H-NMR (D20) S: 2,40 (s, 3H, CH3C00), 3,37 (s, N-CH3), 1-H-NMR (D 2 O) S: 2.40 (s, 3H, CH 3 CO ), 3.37 (s, N-CH 3 ),
3,39 (s, N-CH3), 3,99 (s); 3.39 (s, N-CH 3 ), 3.99 (s);
UV (H20) Xmaks: 226 (cl3144) nm; UV (H20) Xmax: 226 (cl3144) nm;
Analyse for C^B^^OSBrl: Analysis for C^B^^OSBrl:
Beregnet: C 30,08 H 5,51 N 6,38 Calculated: C 30.08 H 5.51 N 6.38
Funnet: C 30,48 H 5,53 N 6,86 Found: C 30.48 H 5.53 N 6.86
B. l-( 2- merkaptoetvl)- l . 4. 4- trimetvlpiperaziniumblsklorid B. 1-(2-mercaptoetyl)-1. 4. 4-trimethylpiperazinium chloride
En blanding av l-(2-acetyltioetyl)-l,4,4-trimetylpiperazi-niumbromidjodid (1,84 g, 4,19 mmol) og 15 ml 6N saltsyre ble oppvarmet til 57<*>C under en nitrogenatmosfære i 2Vi time. Oppløsningen ble konsentrert under redusert trykk til tørr tilstand. Det faste stoff ble suspendert i 10 ml vann og den godt omrørte suspensjon ble behandlet med Permutitt S-l Cl<e >inntil det ble oppnådd oppløsning. Oppløsningen ble helt på en 1,2 x 60 cm kolonne av Permutitt S-l Cl<®>. Kolonnen ble fortynnet med vann (1,5 ml/min.). De passende fraksjoner ble forenet og lyofilisert, noe som ga et hvitt pulver i en mengde av 0,93 g og med smeltepunkt 190-191°C, 85*. A mixture of 1-(2-acetylthioethyl)-1,4,4-trimethylpiperazinium bromide iodide (1.84 g, 4.19 mmol) and 15 ml of 6N hydrochloric acid was heated to 57<*>C under a nitrogen atmosphere for 2Vi h . The solution was concentrated under reduced pressure to dryness. The solid was suspended in 10 ml of water and the well-stirred suspension was treated with Permutitt S-1Cl<e> until dissolution was obtained. The solution was poured onto a 1.2 x 60 cm column of Permutitt S-1 Cl<®>. The column was diluted with water (1.5 ml/min). The appropriate fractions were combined and lyophilized to give a white powder in the amount of 0.93 g and melting point 190-191°C, 85*.
IR (nujol) vmaks: 2460 (SH); IR (nujol) vmax: 2460 (SH);
<1->H-NMR (D20) S: 3,4 (s, N-CH3), 3,45 (s, N-CH3), 4,07 (s); <1->H-NMR (D 2 O) S: 3.4 (s, N-CH 3 ), 3.45 (s, N-CH 3 ), 4.07 (s);
Analyse for CgH22N2SCl2* 0,75H20: Analysis for CgH22N2SCl2* 0.75H20:
Beregnet: C 39,34 H 8,62 N 10,20 S 11,67 Calculated: C 39.34 H 8.62 N 10.20 S 11.67
Funnet: C 39,48 H 8,39 N 10,55 S 11,15 Found: C 39.48 H 8.39 N 10.55 S 11.15
C. ( 5R. 6S)- p- nitrobenzvl- 3- r2-( l. 4. 4- trimetvl- l- Dipera-zlnium ) etvltlo1- 6- riR- hvdroksvetvll- 7- okso- 1- aza-bicvklof 3. 2. 0) hept- 2- en- 2- karboksvlatblsklorid C. ( 5R. 6S)- p- nitrobenzvl- 3- r2-( l. 4. 4- trimethvl- l- Dipera-zlnium ) etvltlo1- riR- hvdroksvetvll- 7- oxo- 1- aza-bicvklof 3. 2. 0) hept- 2- en- 2- carboxyl chloride
Til en kald (5'C) oppløsning av (5R,6S)-p-nitrobenzyl-6-[lR-hydroksyetyl]-3,7-diokso-l-azabicyklo(3.2.0)heptan-2R-karboksylat (0,94 g, 2,7 mmol) i 3 ml acetonitril, holdt under nitrogen, ble det satt diisopropyletylamin (0,557 ml, 3,2 mmol) og difenylklorfosfat (0,663 ml, 3,2 mmol). Reaksjonsblandingen ble omrørt i 30 minutter ved 5°C og behandlet med diisopropyletylamin (0,599 ml, 3,44 mmol) og 4 ml av en vandig oppløsning av l-(2-merkaptoetyl)-l,4,4-trimetylpiperaziniumbisklorid (0,90 g, 3,44 mmol). Efter 1,25 timer ble det tilsatt diisopropyletylamin (0,1 ml, 0,57 mmol) og omrøringen fortsatte i 2 timer. En del av acetonitrilet ble fjernet under redusert trykk og den resulterende røde blanding ble kromatografert på en PrepPAK -500/Cig-søyle (Waters Associates) med 25-75* acetonitril i vann som elueringsmiddel, noe som ga 1,4 g av et gulaktig pulver efter lyofilisering. Dette ble oppløst i vann og oppløsningen helt på en 1,2 x 58 cm kolonne av Permutitt S-l Cl<®> under anvendelse av vann som elueringsmiddel. Lyofilisering av de passende fraksjoner ga 1,17 g av et pulver som på ny ble renset på en PrepPAK -500/Cig-kolonne. Lyofilisering av de passende fraksjoner ga et gulaktig pulver, 0,80 g (53*). To a cold (5'C) solution of (5R,6S)-p-nitrobenzyl-6-[1R-hydroxyethyl]-3,7-dioxo-1-azabicyclo(3.2.0)heptane-2R-carboxylate (0, 94 g, 2.7 mmol) in 3 mL of acetonitrile, kept under nitrogen, was added diisopropylethylamine (0.557 mL, 3.2 mmol) and diphenylchlorophosphate (0.663 mL, 3.2 mmol). The reaction mixture was stirred for 30 minutes at 5°C and treated with diisopropylethylamine (0.599 mL, 3.44 mmol) and 4 mL of an aqueous solution of 1-(2-mercaptoethyl)-1,4,4-trimethylpiperazinium bichloride (0.90 g, 3.44 mmol). After 1.25 hours, diisopropylethylamine (0.1 mL, 0.57 mmol) was added and stirring continued for 2 hours. A portion of the acetonitrile was removed under reduced pressure and the resulting red mixture was chromatographed on a PrepPAK -500/Cig column (Waters Associates) eluting with 25-75% acetonitrile in water to give 1.4 g of a yellowish powder after lyophilization. This was dissolved in water and the solution poured onto a 1.2 x 58 cm column of Permutitt S-1 Cl<®> using water as eluent. Lyophilization of the appropriate fractions gave 1.17 g of a powder which was purified again on a PrepPAK -500/Cig column. Lyophilization of the appropriate fractions gave a yellowish powder, 0.80 g (53*).
IR (KBr) vmaks: 3400 (br, OH), 1770 (C-0 av p-laktam), 1690 IR (KBr) vmax: 3400 (br, OH), 1770 (C-0 of β-lactam), 1690
(C-0 av pNB-ester), 1605 (aromatisk), 1515 (N02), 1345 (N02) cm-<1>; (C-0 of pNB ester), 1605 (aromatic), 1515 (NO2), 1345 (NO2) cm-<1>;
<1->H-NMR (D20) S: 1,26 (d, J-6,3 Hz, 3H, CH3CH0H), 3,39 (s, <1->H-NMR (D2O) S: 1.26 (d, J-6.3 Hz, 3H, CH3CH0H), 3.39 (s,
NCH3), 4,00 (s), 5,37 (br, s, CH2 av pNB), 7,60 (d, J-8,6 Hz, 2H, Ho av pNB), 8,20 (d, J-8,7 Hz, 2H, Hm av pNB); NCH3), 4.00 (s), 5.37 (br, s, CH2 of pNB), 7.60 (d, J-8.6 Hz, 2H, Ho of pNB), 8.20 (d, J -8.7 Hz, 2H, Hm of pNB);
UV (H20) Xmaks: 276 (cl2094), 306 (cl0752) nm; UV (H 2 O) Xmax: 276 (cl2094), 306 (cl0752) nm;
Analyse for C25H3(,N406SC12<*>3H20: Analysis for C25H3(,N406SC12<*>3H20:
Beregnet: C 46,51 H 6,56 N 8,68 S 4,97 Cl 10,98 Funnet: C 46,31 H 6,18 N 8,57 S 5,36 Cl 11,37 D. ( 5R. 6S)- 3- T2-( l. 4. 4- trimetyl- l- plperazinium) etvltiol- 6-riR- hvdroksvetyll- 7- okso- l- azabicyklo( 3. 2. 0 ) hept- 2- en- 2-karboksvlatklorld Calculated: C 46.51 H 6.56 N 8.68 S 4.97 Cl 10.98 Found: C 46.31 H 6.18 N 8.57 S 5.36 Cl 11.37 D. ( 5R. 6S )- 3- T2-( 1. 4. 4- trimethyl- 1- plperazinium) ethylthiol- 6-riR- hydroxybutyl- 7- oxo- 1- azabicyclo( 3. 2. 0 ) hept- 2- ene- 2-carboxvlatechlorld
En blanding av (5R,6S)-p-nitrobenzyl-3-[2-(l,4 ,4-trimetyl-l-piperazinium)etyltio]-6-[lR-hydroksyetyl]-7-okso-l-azabicyklo(3.2.0)hept-2-en-2-karboksylatbisklorid (0,40 g, 0,68 mmol), fosfatbuffer (30 ml, 0,05 M, pH 7,0), tetrahydrofuran (10 ml), eter (30 ml) og 0,40 g 10* palladium-på-trekull ble hydrogenert ved 23°C under et trykk på 2,45 kp/cm<2> i 1 time. De to faser ble separert. Den organiske fase ble ekstrahert med 10 ml vann. De vandige faser ble filtrert på en Celite-pute, vasket med 10 ml eter, konsentrert under vakuum til 10 ml og kromatografert på en PrepPAK -500/Cjg-kolonne (2,2 x 11 cm) med vann som elueringsmiddel, noe som ga 70 mg eller 25* efter lyofilisering; A mixture of (5R,6S)-p-nitrobenzyl-3-[2-(1,4,4-trimethyl-1-piperazinium)ethylthio]-6-[1R-hydroxyethyl]-7-oxo-1-azabicyclo( 3.2.0)hept-2-ene-2-carboxylate bichloride (0.40 g, 0.68 mmol), phosphate buffer (30 mL, 0.05 M, pH 7.0), tetrahydrofuran (10 mL), ether (30 ml) and 0.40 g of 10* palladium-on-charcoal were hydrogenated at 23°C under a pressure of 2.45 kp/cm<2> for 1 hour. The two phases were separated. The organic phase was extracted with 10 ml of water. The aqueous phases were filtered on a Celite pad, washed with 10 mL of ether, concentrated under vacuum to 10 mL and chromatographed on a PrepPAK -500/Cjg column (2.2 x 11 cm) eluting with water to give 70 mg or 25* after lyophilization;
IR (KBr) vmaks: 3400 (br, OH), 1755 (C-0 av e-laktam), 1585 IR (KBr) vmax: 3400 (br, OH), 1755 (C-0 of ε-lactam), 1585
(karboksylat) cm-<*>; (carboxylate) cm-<*>;
*H-NMR (D20) 5: 1,24 (3H, d, J-6,3 Hz, CH3CH0H), 3,36 (s,, NCH3), 3,98 (s); *H-NMR (D 2 O) δ: 1.24 (3H, d, J-6.3 Hz, CH 3 CH 0 H), 3.36 (s, NCH 3 ), 3.98 (s);
UV (H20) Xmaks1 296 (c7987) nm; UV (H 2 O) Xmax1 296 (c7987) nm;
[a]g<3> 35,9<*> (c 0,30, H20); [a]g<3> 35.9<*> (c 0.30, H 2 O);
tj£ - 9,8 timer (målt ved en konsentrasjon på IO-<4> M i fosfatbuffer, pH 7,4, ved 36,8<*>C). tj£ - 9.8 hours (measured at a concentration of 10-<4> M in phosphate buffer, pH 7.4, at 36.8<*>C).
Claims (1)
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NO902554A NO171271C (en) | 1983-03-08 | 1990-06-08 | QUATERNARY AMINTIOL COMPOUNDS |
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US47137883A | 1983-03-08 | 1983-03-08 | |
US49969183A | 1983-06-07 | 1983-06-07 | |
US55729583A | 1983-12-01 | 1983-12-01 | |
NO840865A NO166038C (en) | 1983-03-08 | 1984-03-07 | PROCEDURE FOR THE PREPARATION OF CARBAPENEM DERIVATIVES. |
NO902554A NO171271C (en) | 1983-03-08 | 1990-06-08 | QUATERNARY AMINTIOL COMPOUNDS |
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Publication Number | Publication Date |
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NO902554L NO902554L (en) | 1984-09-10 |
NO902554D0 NO902554D0 (en) | 1990-06-08 |
NO171271B true NO171271B (en) | 1992-11-09 |
NO171271C NO171271C (en) | 1993-02-17 |
Family
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NO902554A NO171271C (en) | 1983-03-08 | 1990-06-08 | QUATERNARY AMINTIOL COMPOUNDS |
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NO902554D0 (en) | 1990-06-08 |
NO902554L (en) | 1984-09-10 |
NO171271C (en) | 1993-02-17 |
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