WO1996027379A9 - Carbopeptoids and carbonucleotoids - Google Patents
Carbopeptoids and carbonucleotoidsInfo
- Publication number
- WO1996027379A9 WO1996027379A9 PCT/US1996/003227 US9603227W WO9627379A9 WO 1996027379 A9 WO1996027379 A9 WO 1996027379A9 US 9603227 W US9603227 W US 9603227W WO 9627379 A9 WO9627379 A9 WO 9627379A9
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- equivalents
- carbon
- carbohydrate
- anomeric
- solution
- Prior art date
Links
- 150000001720 carbohydrates Chemical class 0.000 claims abstract description 21
- 150000001875 compounds Chemical class 0.000 claims description 160
- -1 carbohydrate amino acid Chemical class 0.000 claims description 102
- OKTJSMMVPCPJKN-UHFFFAOYSA-N carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 64
- 229910052799 carbon Inorganic materials 0.000 claims description 57
- 238000000034 method Methods 0.000 claims description 51
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 42
- 230000002194 synthesizing Effects 0.000 claims description 42
- 150000001408 amides Chemical class 0.000 claims description 36
- 229910052739 hydrogen Inorganic materials 0.000 claims description 34
- 150000004713 phosphodiesters Chemical class 0.000 claims description 27
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 19
- 239000001257 hydrogen Substances 0.000 claims description 16
- VILAVOFMIJHSJA-UHFFFAOYSA-N Dicarbon monoxide Chemical compound [C]=C=O VILAVOFMIJHSJA-UHFFFAOYSA-N 0.000 claims description 14
- 150000000700 C-glycosides Chemical class 0.000 claims description 12
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Substances N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- 229910052757 nitrogen Inorganic materials 0.000 claims description 11
- 150000002431 hydrogen Chemical class 0.000 claims description 8
- 230000001808 coupling Effects 0.000 claims description 6
- 238000010168 coupling process Methods 0.000 claims description 6
- 238000005859 coupling reaction Methods 0.000 claims description 6
- 238000007877 drug screening Methods 0.000 claims description 4
- ORTFAQDWJHRMNX-UHFFFAOYSA-M oxidooxomethyl Chemical compound [O-][C]=O ORTFAQDWJHRMNX-UHFFFAOYSA-M 0.000 claims description 3
- AQSJGOWTSHOLKH-UHFFFAOYSA-N Phosphite Chemical group [O-]P([O-])[O-] AQSJGOWTSHOLKH-UHFFFAOYSA-N 0.000 claims 2
- 229920001542 oligosaccharide Polymers 0.000 abstract description 27
- 150000002482 oligosaccharides Polymers 0.000 abstract description 27
- 229920000023 polynucleotide Polymers 0.000 abstract 1
- 239000002157 polynucleotide Substances 0.000 abstract 1
- 229920001184 polypeptide Polymers 0.000 abstract 1
- RTZKZFJDLAIYFH-UHFFFAOYSA-N diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 322
- 239000000243 solution Substances 0.000 description 191
- YMWUJEATGCHHMB-UHFFFAOYSA-N methylene dichloride Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 162
- 239000012267 brine Substances 0.000 description 141
- 238000006243 chemical reaction Methods 0.000 description 138
- CSNNHWWHGAXBCP-UHFFFAOYSA-L mgso4 Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 133
- XEKOWRVHYACXOJ-UHFFFAOYSA-N acetic acid ethyl ester Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 126
- 238000003818 flash chromatography Methods 0.000 description 126
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 126
- 238000002360 preparation method Methods 0.000 description 123
- 238000000746 purification Methods 0.000 description 121
- QDHHCQZDFGDHMP-UHFFFAOYSA-N monochloramine Chemical compound ClN QDHHCQZDFGDHMP-UHFFFAOYSA-N 0.000 description 111
- UIIMBOGNXHQVGW-UHFFFAOYSA-M buffer Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 80
- 239000000203 mixture Substances 0.000 description 75
- 229910001868 water Inorganic materials 0.000 description 72
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 71
- 238000003756 stirring Methods 0.000 description 66
- OKKJLVBELUTLKV-UHFFFAOYSA-N methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 56
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 55
- WYURNTSHIVDZCO-UHFFFAOYSA-N tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 53
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonium chloride Substances [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 52
- 235000019270 ammonium chloride Nutrition 0.000 description 50
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 50
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 48
- 229960004132 diethyl ether Drugs 0.000 description 48
- 235000014633 carbohydrates Nutrition 0.000 description 46
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 40
- 239000011541 reaction mixture Substances 0.000 description 39
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 38
- JUJWROOIHBZHMG-UHFFFAOYSA-N pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 36
- HEMHJVSKTPXQMS-UHFFFAOYSA-M sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 33
- 238000007792 addition Methods 0.000 description 31
- 239000002253 acid Substances 0.000 description 29
- 230000015572 biosynthetic process Effects 0.000 description 26
- 238000003786 synthesis reaction Methods 0.000 description 26
- IRXSLJNXXZKURP-UHFFFAOYSA-N Fluorenylmethyloxycarbonyl chloride Chemical compound C1=CC=C2C(COC(=O)Cl)C3=CC=CC=C3C2=C1 IRXSLJNXXZKURP-UHFFFAOYSA-N 0.000 description 24
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 21
- 239000011521 glass Substances 0.000 description 21
- 239000000377 silicon dioxide Substances 0.000 description 21
- 239000007787 solid Substances 0.000 description 20
- 125000003277 amino group Chemical group 0.000 description 18
- 239000012044 organic layer Substances 0.000 description 17
- 235000017557 sodium bicarbonate Nutrition 0.000 description 17
- YYROPELSRYBVMQ-UHFFFAOYSA-N 4-Toluenesulfonyl chloride Chemical compound CC1=CC=C(S(Cl)(=O)=O)C=C1 YYROPELSRYBVMQ-UHFFFAOYSA-N 0.000 description 16
- VZGDMQKNWNREIO-UHFFFAOYSA-N Carbon tetrachloride Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 16
- 239000003208 petroleum Substances 0.000 description 16
- 239000000126 substance Substances 0.000 description 16
- 239000000543 intermediate Substances 0.000 description 15
- 239000012047 saturated solution Substances 0.000 description 15
- 229910019891 RuCl3 Inorganic materials 0.000 description 14
- YBCAZPLXEGKKFM-UHFFFAOYSA-K Ruthenium(III) chloride Chemical compound [Cl-].[Cl-].[Cl-].[Ru+3] YBCAZPLXEGKKFM-UHFFFAOYSA-K 0.000 description 14
- 239000000047 product Substances 0.000 description 14
- STMPXDBGVJZCEX-UHFFFAOYSA-N triethylsilyl trifluoromethanesulfonate Chemical compound CC[Si](CC)(CC)OS(=O)(=O)C(F)(F)F STMPXDBGVJZCEX-UHFFFAOYSA-N 0.000 description 14
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 13
- 239000010410 layer Substances 0.000 description 13
- 239000012074 organic phase Substances 0.000 description 13
- LEIMLDGFXIOXMT-UHFFFAOYSA-N Trimethylsilyl cyanide Chemical compound C[Si](C)(C)C#N LEIMLDGFXIOXMT-UHFFFAOYSA-N 0.000 description 12
- HPQVWDOOUQVBTO-UHFFFAOYSA-N lithium aluminium hydride Substances [Li+].[Al-] HPQVWDOOUQVBTO-UHFFFAOYSA-N 0.000 description 12
- VMHLLURERBWHNL-UHFFFAOYSA-M sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 12
- 239000001632 sodium acetate Substances 0.000 description 12
- 235000017281 sodium acetate Nutrition 0.000 description 12
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N sodium azide Substances [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 12
- ZMANZCXQSJIPKH-UHFFFAOYSA-N triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 12
- ARUVKPQLZAKDPS-UHFFFAOYSA-L Copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 11
- 108010038807 Oligopeptides Proteins 0.000 description 11
- 102000015636 Oligopeptides Human genes 0.000 description 11
- KDLHZDBZIXYQEI-UHFFFAOYSA-N Palladium on carbon Substances [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 11
- 150000007513 acids Chemical class 0.000 description 11
- 229910000365 copper sulfate Inorganic materials 0.000 description 11
- OISVCGZHLKNMSJ-UHFFFAOYSA-N 2,6-Lutidine Chemical compound CC1=CC=CC(C)=N1 OISVCGZHLKNMSJ-UHFFFAOYSA-N 0.000 description 10
- JVVRCYWZTJLJSG-UHFFFAOYSA-N 4-Dimethylaminophenol Substances CN(C)C1=CC=C(O)C=C1 JVVRCYWZTJLJSG-UHFFFAOYSA-N 0.000 description 10
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 10
- MKRTXPORKIRPDG-UHFFFAOYSA-N Diphenylphosphoryl azide Chemical compound C=1C=CC=CC=1P(=O)(N=[N+]=[N-])C1=CC=CC=C1 MKRTXPORKIRPDG-UHFFFAOYSA-N 0.000 description 10
- 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 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 10
- 239000003153 chemical reaction reagent Substances 0.000 description 10
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 10
- GZCGUPFRVQAUEE-KCDKBNATSA-N D-(+)-Galactose Natural products OC[C@@H](O)[C@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-KCDKBNATSA-N 0.000 description 9
- 108010043958 Peptoids Proteins 0.000 description 9
- 150000001412 amines Chemical class 0.000 description 9
- 239000010452 phosphate Substances 0.000 description 9
- 238000010791 quenching Methods 0.000 description 9
- 238000010992 reflux Methods 0.000 description 9
- 239000007858 starting material Substances 0.000 description 9
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 8
- KJUGUADJHNHALS-UHFFFAOYSA-N 1H-tetrazole Chemical compound C=1N=NNN=1 KJUGUADJHNHALS-UHFFFAOYSA-N 0.000 description 8
- YEDUAINPPJYDJZ-UHFFFAOYSA-N 2-hydroxybenzothiazole Chemical compound C1=CC=C2SC(O)=NC2=C1 YEDUAINPPJYDJZ-UHFFFAOYSA-N 0.000 description 8
- HUMNYLRZRPPJDN-UHFFFAOYSA-N Benzaldehyde Chemical compound O=CC1=CC=CC=C1 HUMNYLRZRPPJDN-UHFFFAOYSA-N 0.000 description 8
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Carbodicyclohexylimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N D-Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 8
- NHQDETIJWKXCTC-UHFFFAOYSA-N Meta-Chloroperoxybenzoic acid Chemical compound OOC(=O)C1=CC=CC(Cl)=C1 NHQDETIJWKXCTC-UHFFFAOYSA-N 0.000 description 8
- LYGJENNIWJXYER-UHFFFAOYSA-N Nitromethane Chemical compound C[N+]([O-])=O LYGJENNIWJXYER-UHFFFAOYSA-N 0.000 description 8
- JQWHASGSAFIOCM-UHFFFAOYSA-M Sodium periodate Chemical compound [Na+].[O-]I(=O)(=O)=O JQWHASGSAFIOCM-UHFFFAOYSA-M 0.000 description 8
- UQDAEORWFCPQCU-UHFFFAOYSA-N acetic acid;oxolane;hydrate Chemical compound O.CC(O)=O.C1CCOC1 UQDAEORWFCPQCU-UHFFFAOYSA-N 0.000 description 8
- UHOVQNZJYSORNB-UHFFFAOYSA-N benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-M chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 8
- 239000006185 dispersion Substances 0.000 description 8
- UFHFLCQGNIYNRP-UHFFFAOYSA-N hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 8
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 8
- 239000002480 mineral oil Substances 0.000 description 8
- 235000010446 mineral oil Nutrition 0.000 description 8
- NQRYJNQNLNOLGT-UHFFFAOYSA-N piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 8
- VVWRJUBEIPHGQF-MDZDMXLPSA-N propan-2-yl (NE)-N-propan-2-yloxycarbonyliminocarbamate Chemical compound CC(C)OC(=O)\N=N\C(=O)OC(C)C VVWRJUBEIPHGQF-MDZDMXLPSA-N 0.000 description 8
- BZKBCQXYZZXSCO-UHFFFAOYSA-N sodium hydride Inorganic materials [H-].[Na+] BZKBCQXYZZXSCO-UHFFFAOYSA-N 0.000 description 8
- WQDUMFSSJAZKTM-UHFFFAOYSA-N sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 description 8
- 229960001031 Glucose Drugs 0.000 description 7
- LJCNRYVRMXRIQR-UHFFFAOYSA-L Potassium sodium tartrate Chemical compound [Na+].[K+].[O-]C(=O)C(O)C(O)C([O-])=O LJCNRYVRMXRIQR-UHFFFAOYSA-L 0.000 description 7
- 125000000738 acetamido group Chemical group [H]C([H])([H])C(=O)N([H])[*] 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 7
- 125000000649 benzylidene group Chemical group [H]C(=[*])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 7
- 238000010276 construction Methods 0.000 description 7
- 239000012043 crude product Substances 0.000 description 7
- 238000010790 dilution Methods 0.000 description 7
- 239000000706 filtrate Substances 0.000 description 7
- 125000001810 isothiocyanato group Chemical group *N=C=S 0.000 description 7
- 125000005647 linker group Chemical group 0.000 description 7
- 235000019341 magnesium sulphate Nutrition 0.000 description 7
- ZXDUPDQEFOYLOM-UHFFFAOYSA-O propylideneazanium Chemical group [CH2-]CC=[NH2+] ZXDUPDQEFOYLOM-UHFFFAOYSA-O 0.000 description 7
- 229920005989 resin Polymers 0.000 description 7
- 239000011347 resin Substances 0.000 description 7
- 239000001476 sodium potassium tartrate Substances 0.000 description 7
- 235000011006 sodium potassium tartrate Nutrition 0.000 description 7
- 238000005406 washing Methods 0.000 description 7
- WQZGKKKJIJFFOK-VFUOTHLCSA-N β-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 7
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-dioxane Chemical class C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 6
- JVSFQJZRHXAUGT-UHFFFAOYSA-N 2,2-dimethylpropanoyl chloride Chemical compound CC(C)(C)C(Cl)=O JVSFQJZRHXAUGT-UHFFFAOYSA-N 0.000 description 6
- QWTBDIBOOIAZEF-UHFFFAOYSA-N 3-[chloro-[di(propan-2-yl)amino]phosphanyl]oxypropanenitrile Chemical compound CC(C)N(C(C)C)P(Cl)OCCC#N QWTBDIBOOIAZEF-UHFFFAOYSA-N 0.000 description 6
- AGEZXYOZHKGVCM-UHFFFAOYSA-N Benzyl bromide Chemical compound BrCC1=CC=CC=C1 AGEZXYOZHKGVCM-UHFFFAOYSA-N 0.000 description 6
- VZJVWSHVAAUDKD-UHFFFAOYSA-N Potassium permanganate Chemical compound [K+].[O-][Mn](=O)(=O)=O VZJVWSHVAAUDKD-UHFFFAOYSA-N 0.000 description 6
- WFDIJRYMOXRFFG-UHFFFAOYSA-N acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 6
- 125000000217 alkyl group Chemical group 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 6
- 230000027455 binding Effects 0.000 description 6
- AZWXAPCAJCYGIA-UHFFFAOYSA-N bis(2-methylpropyl)alumane Chemical compound CC(C)C[AlH]CC(C)C AZWXAPCAJCYGIA-UHFFFAOYSA-N 0.000 description 6
- CPELXLSAUQHCOX-UHFFFAOYSA-M bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 6
- 239000003921 oil Substances 0.000 description 6
- 229920005990 polystyrene resin Polymers 0.000 description 6
- 229940086735 succinate Drugs 0.000 description 6
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 6
- DKGAVHZHDRPRBM-UHFFFAOYSA-N t-BuOH Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 description 6
- 229940086542 triethylamine Drugs 0.000 description 6
- DTQVDTLACAAQTR-UHFFFAOYSA-N trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 6
- 238000005160 1H NMR spectroscopy Methods 0.000 description 5
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 5
- 229940077731 Carbohydrate nutrients Drugs 0.000 description 5
- 229960002442 Glucosamine Drugs 0.000 description 5
- 229920000272 Oligonucleotide Polymers 0.000 description 5
- HPGGPRDJHPYFRM-UHFFFAOYSA-J Tin(IV) chloride Chemical compound Cl[Sn](Cl)(Cl)Cl HPGGPRDJHPYFRM-UHFFFAOYSA-J 0.000 description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-K [O-]P([O-])([O-])=O Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 5
- WPYMKLBDIGXBTP-UHFFFAOYSA-M benzoate Chemical compound [O-]C(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-M 0.000 description 5
- MHDVGSVTJDSBDK-UHFFFAOYSA-N dibenzyl ether Chemical compound C=1C=CC=CC=1COCC1=CC=CC=C1 MHDVGSVTJDSBDK-UHFFFAOYSA-N 0.000 description 5
- 239000008103 glucose Substances 0.000 description 5
- 150000008300 phosphoramidites Chemical group 0.000 description 5
- 229910000027 potassium carbonate Inorganic materials 0.000 description 5
- 125000000037 tert-butyldiphenylsilyl group Chemical group [H]C1=C([H])C([H])=C([H])C([H])=C1[Si]([H])([*]C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 5
- 150000003536 tetrazoles Chemical class 0.000 description 5
- QXTIBZLKQPJVII-UHFFFAOYSA-N triethylsilicon Chemical compound CC[Si](CC)CC QXTIBZLKQPJVII-UHFFFAOYSA-N 0.000 description 5
- 150000004072 triols Chemical class 0.000 description 5
- NTIXPPFPXLYJCT-CFTHBVFTSA-N (3S,5R,7S,8R,9S,10S,12R,13R,14S,17R)-17-[(2R)-6-hydroxy-6-methylheptan-2-yl]-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3,7,12-triol Chemical compound C([C@@H]1C[C@@H]2O)[C@@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCCC(C)(C)O)C)[C@@]2(C)[C@H](O)C1 NTIXPPFPXLYJCT-CFTHBVFTSA-N 0.000 description 4
- UHOVQNZJYSORNB-MZWXYZOWSA-N Deuterated benzene Chemical compound [2H]C1=C([2H])C([2H])=C([2H])C([2H])=C1[2H] UHOVQNZJYSORNB-MZWXYZOWSA-N 0.000 description 4
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-Acetylglucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 description 4
- 229950006780 N-Acetylglucosamine Drugs 0.000 description 4
- FPGGTKZVZWFYPV-UHFFFAOYSA-M Tetra-n-butylammonium fluoride Chemical compound [F-].CCCC[N+](CCCC)(CCCC)CCCC FPGGTKZVZWFYPV-UHFFFAOYSA-M 0.000 description 4
- 239000011324 bead Substances 0.000 description 4
- 229940095076 benzaldehyde Drugs 0.000 description 4
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 4
- 230000001413 cellular Effects 0.000 description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- AXFQBNWFPNLNJB-UHFFFAOYSA-L dichloro(diethyl)stannane;1,10-phenanthroline Chemical compound CC[Sn](Cl)(Cl)CC.C1=CN=C2C3=NC=CC=C3C=CC2=C1 AXFQBNWFPNLNJB-UHFFFAOYSA-L 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N formic acid Chemical compound OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 235000019253 formic acid Nutrition 0.000 description 4
- 230000001590 oxidative Effects 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 235000000346 sugar Nutrition 0.000 description 4
- LPTITAGPBXDDGR-IBEHDNSVSA-N 2,3,4,5,6-Penta-O-acetyl-D-glucose Chemical compound CC(=O)OC[C@H]1O[C@@H](OC(C)=O)[C@H](OC(C)=O)[C@@H](OC(C)=O)[C@@H]1OC(C)=O LPTITAGPBXDDGR-IBEHDNSVSA-N 0.000 description 3
- MTJGVAJYTOXFJH-UHFFFAOYSA-N 3-aminonaphthalene-1,5-disulfonic acid Chemical compound C1=CC=C(S(O)(=O)=O)C2=CC(N)=CC(S(O)(=O)=O)=C21 MTJGVAJYTOXFJH-UHFFFAOYSA-N 0.000 description 3
- 229910021012 Co2(CO)8 Inorganic materials 0.000 description 3
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 3
- BEOOHQFXGBMRKU-UHFFFAOYSA-N Sodium cyanoborohydride Chemical compound [Na+].[B-]C#N BEOOHQFXGBMRKU-UHFFFAOYSA-N 0.000 description 3
- QDRKDTQENPPHOJ-UHFFFAOYSA-N Sodium ethoxide Chemical compound [Na+].CC[O-] QDRKDTQENPPHOJ-UHFFFAOYSA-N 0.000 description 3
- 230000000975 bioactive Effects 0.000 description 3
- BXCQGSQPWPGFIV-UHFFFAOYSA-N carbon monoxide;cobalt;cobalt(2+);methanone Chemical compound [Co].[Co+2].O=[CH-].O=[CH-].[O+]#[C-].[O+]#[C-].[O+]#[C-].[O+]#[C-].[O+]#[C-].[O+]#[C-] BXCQGSQPWPGFIV-UHFFFAOYSA-N 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 3
- 229940000406 drug candidates Drugs 0.000 description 3
- 238000005755 formation reaction Methods 0.000 description 3
- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 3
- 239000002808 molecular sieve Substances 0.000 description 3
- 238000006384 oligomerization reaction Methods 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 125000006239 protecting group Chemical group 0.000 description 3
- 239000007790 solid phase Substances 0.000 description 3
- JOXIMZWYDAKGHI-UHFFFAOYSA-M toluene-4-sulfonate Chemical compound CC1=CC=C(S([O-])(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-M 0.000 description 3
- QKPLRMLTKYXDST-BMZZJELJSA-N (3R,4R,5R,6R)-3-amino-6-(hydroxymethyl)oxane-2,4,5-triol;hydron;chloride Chemical compound Cl.N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O QKPLRMLTKYXDST-BMZZJELJSA-N 0.000 description 2
- MSWZFWKMSRAUBD-UHFFFAOYSA-N 3-amino-6-(hydroxymethyl)oxane-2,4,5-triol Chemical compound NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 2
- AYJRCSIUFZENHW-UHFFFAOYSA-L Barium carbonate Chemical compound [Ba+2].[O-]C([O-])=O AYJRCSIUFZENHW-UHFFFAOYSA-L 0.000 description 2
- CCWPXMLXXAIAJP-UHFFFAOYSA-N CCCC[Sn](CCCC)=CC1=CC=CC=C1 Chemical compound CCCC[Sn](CCCC)=CC1=CC=CC=C1 CCWPXMLXXAIAJP-UHFFFAOYSA-N 0.000 description 2
- FVIGODVHAVLZOO-UHFFFAOYSA-N Dixanthogen Chemical compound CCOC(=S)SSC(=S)OCC FVIGODVHAVLZOO-UHFFFAOYSA-N 0.000 description 2
- RAXXELZNTBOGNW-UHFFFAOYSA-N Imidazole Chemical compound C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 2
- 208000007976 Ketosis Diseases 0.000 description 2
- 239000002841 Lewis acid Substances 0.000 description 2
- PCLIMKBDDGJMGD-UHFFFAOYSA-N N-Bromosuccinimide Chemical compound BrN1C(=O)CCC1=O PCLIMKBDDGJMGD-UHFFFAOYSA-N 0.000 description 2
- 238000006987 Nef reaction Methods 0.000 description 2
- LGRFSURHDFAFJT-UHFFFAOYSA-N Phthalic anhydride Chemical compound C1=CC=C2C(=O)OC(=O)C2=C1 LGRFSURHDFAFJT-UHFFFAOYSA-N 0.000 description 2
- 239000001083 [(2R,3R,4S,5R)-1,2,4,5-tetraacetyloxy-6-oxohexan-3-yl] acetate Substances 0.000 description 2
- UAOKXEHOENRFMP-ZJIFWQFVSA-N [(2R,3R,4S,5R)-2,3,4,5-tetraacetyloxy-6-oxohexyl] acetate Chemical compound CC(=O)OC[C@@H](OC(C)=O)[C@@H](OC(C)=O)[C@H](OC(C)=O)[C@@H](OC(C)=O)C=O UAOKXEHOENRFMP-ZJIFWQFVSA-N 0.000 description 2
- LPTITAGPBXDDGR-LYYZXLFJSA-N [(2R,3S,4S,5R,6S)-3,4,5,6-tetraacetyloxyoxan-2-yl]methyl acetate Chemical compound CC(=O)OC[C@H]1O[C@@H](OC(C)=O)[C@H](OC(C)=O)[C@@H](OC(C)=O)[C@H]1OC(C)=O LPTITAGPBXDDGR-LYYZXLFJSA-N 0.000 description 2
- 230000002378 acidificating Effects 0.000 description 2
- PYMYPHUHKUWMLA-LMVFSUKVSA-N aldehydo-D-ribose Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 2
- AQIHMSVIAGNIDM-UHFFFAOYSA-N benzoyl bromide Chemical compound BrC(=O)C1=CC=CC=C1 AQIHMSVIAGNIDM-UHFFFAOYSA-N 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- UWTDFICHZKXYAC-UHFFFAOYSA-N boron;oxolane Chemical compound [B].C1CCOC1 UWTDFICHZKXYAC-UHFFFAOYSA-N 0.000 description 2
- XJHCXCQVJFPJIK-UHFFFAOYSA-M caesium fluoride Inorganic materials [F-].[Cs+] XJHCXCQVJFPJIK-UHFFFAOYSA-M 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 230000035569 catabolism Effects 0.000 description 2
- 159000000006 cesium salts Chemical class 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 235000008504 concentrate Nutrition 0.000 description 2
- 230000000875 corresponding Effects 0.000 description 2
- 238000006264 debenzylation reaction Methods 0.000 description 2
- YXHKONLOYHBTNS-UHFFFAOYSA-N diazomethane Chemical compound C=[N+]=[N-] YXHKONLOYHBTNS-UHFFFAOYSA-N 0.000 description 2
- 150000002009 diols Chemical class 0.000 description 2
- YBPGMKDTGOJASU-UHFFFAOYSA-N ethanol;2-(oxolan-2-yl)acetic acid Chemical compound CCO.OC(=O)CC1CCCO1 YBPGMKDTGOJASU-UHFFFAOYSA-N 0.000 description 2
- KZMGYPLQYOPHEL-UHFFFAOYSA-N ethoxyethane;trifluoroborane Chemical compound FB(F)F.CCOCC KZMGYPLQYOPHEL-UHFFFAOYSA-N 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 150000002584 ketoses Chemical class 0.000 description 2
- 150000007517 lewis acids Chemical class 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 2
- 235000015320 potassium carbonate Nutrition 0.000 description 2
- GRJJQCWNZGRKAU-UHFFFAOYSA-N pyridin-1-ium;fluoride Chemical compound F.C1=CC=NC=C1 GRJJQCWNZGRKAU-UHFFFAOYSA-N 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- 230000002829 reduced Effects 0.000 description 2
- 150000008163 sugars Chemical group 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000010189 synthetic method Methods 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- MHYGQXWCZAYSLJ-UHFFFAOYSA-N tert-butyl-chloro-diphenylsilane Chemical compound C=1C=CC=CC=1[Si](Cl)(C(C)(C)C)C1=CC=CC=C1 MHYGQXWCZAYSLJ-UHFFFAOYSA-N 0.000 description 2
- 229910001887 tin oxide Inorganic materials 0.000 description 2
- WQZGKKKJIJFFOK-RWOPYEJCSA-N β-D-mannose Chemical compound OC[C@H]1O[C@@H](O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-RWOPYEJCSA-N 0.000 description 2
- WQZGKKKJIJFFOK-RDQKPOQOSA-N (2S,3S,4R,5R,6R)-6-(hydroxymethyl)oxane-2,3,4,5-tetrol Chemical compound OC[C@H]1O[C@H](O)[C@@H](O)[C@H](O)[C@H]1O WQZGKKKJIJFFOK-RDQKPOQOSA-N 0.000 description 1
- WQZGKKKJIJFFOK-TVIMKVIFSA-N (2S,3S,4R,5S,6R)-6-(hydroxymethyl)oxane-2,3,4,5-tetrol Chemical compound OC[C@H]1O[C@H](O)[C@@H](O)[C@H](O)[C@@H]1O WQZGKKKJIJFFOK-TVIMKVIFSA-N 0.000 description 1
- QPFMBZIOSGYJDE-UHFFFAOYSA-N 1,1,2,2-Tetrachloroethane Chemical compound ClC(Cl)C(Cl)Cl QPFMBZIOSGYJDE-UHFFFAOYSA-N 0.000 description 1
- 229920001429 Chelating resin Polymers 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N HCl Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- SRBFZHDQGSBBOR-OWMBCFKOSA-N L-ribopyranose Chemical compound O[C@H]1COC(O)[C@@H](O)[C@H]1O SRBFZHDQGSBBOR-OWMBCFKOSA-N 0.000 description 1
- 101710027499 Os03g0268000 Proteins 0.000 description 1
- 229910018891 PSi Inorganic materials 0.000 description 1
- 235000011158 Prunus mume Nutrition 0.000 description 1
- 240000002546 Prunus mume Species 0.000 description 1
- NDVLTYZPCACLMA-UHFFFAOYSA-N Silver oxide Chemical compound [O-2].[Ag+].[Ag+] NDVLTYZPCACLMA-UHFFFAOYSA-N 0.000 description 1
- 229910008433 SnCU Inorganic materials 0.000 description 1
- 241000282485 Vulpes vulpes Species 0.000 description 1
- 238000005903 acid hydrolysis reaction Methods 0.000 description 1
- GZCGUPFRVQAUEE-SLPGGIOYSA-N aldehydo-D-glucose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-SLPGGIOYSA-N 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 238000010976 amide bond formation reaction Methods 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 239000000908 ammonium hydroxide Substances 0.000 description 1
- 229920001222 biopolymer Polymers 0.000 description 1
- 238000006664 bond formation reaction Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229910021386 carbon form Inorganic materials 0.000 description 1
- UGFAIRIUMAVXCW-UHFFFAOYSA-N carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 description 1
- 229910002091 carbon monoxide Inorganic materials 0.000 description 1
- MSMNPNAQGHVRBA-UHFFFAOYSA-N chloro-[di(propan-2-yl)amino]phosphinite Chemical compound CC(C)N(C(C)C)P([O-])Cl MSMNPNAQGHVRBA-UHFFFAOYSA-N 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000010779 crude oil Substances 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- JQZUMFHYRULBEN-UHFFFAOYSA-N diethyl(methyl)silicon Chemical compound CC[Si](C)CC JQZUMFHYRULBEN-UHFFFAOYSA-N 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- WSFSSNUMVMOOMR-UHFFFAOYSA-N formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 150000002373 hemiacetals Chemical class 0.000 description 1
- 125000005842 heteroatoms Chemical group 0.000 description 1
- 239000000710 homodimer Substances 0.000 description 1
- 229910000041 hydrogen chloride Inorganic materials 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-M hydroxyl anion Chemical compound [OH-] XLYOFNOQVPJJNP-UHFFFAOYSA-M 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000010907 mechanical stirring Methods 0.000 description 1
- 230000001404 mediated Effects 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L na2so4 Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- NSNPSJGHTQIXDO-UHFFFAOYSA-N naphthalene-1-carbonyl chloride Chemical compound C1=CC=C2C(C(=O)Cl)=CC=CC2=C1 NSNPSJGHTQIXDO-UHFFFAOYSA-N 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 238000002515 oligonucleotide synthesis Methods 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 229910052763 palladium Inorganic materials 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- PTMHPRAIXMAOOB-UHFFFAOYSA-L phosphoramidate Chemical compound NP([O-])([O-])=O PTMHPRAIXMAOOB-UHFFFAOYSA-L 0.000 description 1
- 239000001184 potassium carbonate Substances 0.000 description 1
- 229910000108 silver(I,III) oxide Inorganic materials 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- ILMRJRBKQSSXGY-UHFFFAOYSA-N tert-butyl(dimethyl)silicon Chemical compound C[Si](C)C(C)(C)C ILMRJRBKQSSXGY-UHFFFAOYSA-N 0.000 description 1
- 125000001981 tert-butyldimethylsilyl group Chemical group [H]C([H])([H])[Si]([H])(C([H])([H])[H])[*]C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- ILJSQTXMGCGYMG-UHFFFAOYSA-M triacetate(1-) Chemical compound CC(=O)CC(=O)CC([O-])=O ILJSQTXMGCGYMG-UHFFFAOYSA-M 0.000 description 1
- WQZGKKKJIJFFOK-DVKNGEFBSA-N α-D-glucose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-DVKNGEFBSA-N 0.000 description 1
- WQZGKKKJIJFFOK-RXRWUWDJSA-N α-D-gulose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@H](O)[C@H]1O WQZGKKKJIJFFOK-RXRWUWDJSA-N 0.000 description 1
Definitions
- the invention relates to oligosaccharides and libraries incorporating oligosaccharide. More
- the invention relates to oligosaccharides and libraries of oligosaccharides which employ amide and/or phosphodiester linkages for joining adjacent carbohydrate subunits.
- Carbohydrates are known to mediate many cellular recognition processes. Carbohydrates can serve directly as binding molecules and, in such instances, are
- Dysfunctional mediation of cellular recognition processes can lead to disease states. If a cellular recognition process is mediated by an oligosaccharide, then an absence or excess of such oligosaccharide can lead to a dysfunctional mediation of such process.
- the mediating oligosaccharide may be deficient or absent due to a deficiency of production or due to a high rate of catabolism. If rate of catabolism is excessive, then catabolically resistant analogs of the bioactive
- oligosaccharide may be preferred as drug candidates as compared to the native bioactive oligosaccharide .
- a library which includes analogs of known bioactive oligosaccharides .
- Such a library may be usefully employed for screening drug candidates .
- oligopeptoids are shown by calculation to have greater conformational freedom as compared to conventional oligopeptides . Accordingly, oligopeptoids are thought to have greater potential as pharmaceutically useful binding ligands as compared to conventional oligopeptides having close sequence homology to such oligopeptoids .
- Von Roedern et al disclose a carbohydrate amino acid ⁇ Angew. Chem, Int . Ed. Engl . 1994 , 31 , 687-689 ) . Although von Roedern discloses that carbohydrate amino acids may be coupled to peptides , he does not disclose that they may also be polymerized so as to form
- a first aspect of the invention involves the molecular design and chemical synthesis of a class of carbohydrates designated as carbopeptoids (CPD ' s ) .
- Glycopeptoids are preferred carbopeptoids .
- Carbopeptoids and glcopeptoids are oligosaccharides which employ peptide- like amide bonds for linking the various
- the invention is directed to a oligomeric carbopeptoid or glycopeptoid compound having carbohydrate amino acid subunits (CA ' s ) or glycoside amino acid subunits (GA' s ) coupled to one another via an amide linkage .
- the amide linkage may be represented by the formula CA 1 - (CO-NH) -CA 2 .
- the amide linkage (CO-NH) includes a carbonyl carbon and an amido nitrogen .
- a first carbohydrate amino acid subunit CA 1 or glycoside amino acid subunit GA 1 has an anomeric carbon bonded to the carbonyl carbon of the amide linkage .
- the anomeric carbon of the first carbohydrate amino acid subunit CA 1 forms a C-glycosidic bond with the carbonyl carbon of the amide linkage and maintains the carbohydrate in a closed ring configuration .
- a second carbohydrate amino acid subunit CA 2 has a non-anomeric carbon bonded to the amido nitrogen of the amide linkage .
- the second carbohydrate amino acid subunit CA 2 like the first amino acid subunit CA 1 ( may include an anomeric carbon bonded to the
- carbohydrate amino acid subunit CA 2 is a terminal subunit , then its anomeric carbon may form a hemiacetal , a hemiketal , or a glycoside .
- the invention is also directed to a process for synthesizing the above oligomeric carbopeptoid or glycopeptoid compound.
- the synthetic process involves the coupling of two or more carbohydrate amino acid subunits (CA ' s ) or glycoside amino acid subunits (GA' s ) to one another by means of amide linkages .
- the invention is also directed to libraries of oligomeric carbopeptoid or glycopeptoid compounds . Such libraries are employable for drug screening .
- Each oligomeric carbopeptoid or glydopeptoid compound includes at least two carbohydrate amino acid subunits (CA ' s ) or glycoside amino acid subunits (GA' s ) coupled to one another via an amide linkage as indicated above .
- the invention is also directed to an improved process for synthesizing the above library of oligomers .
- the process employs an elongation step for coupling the subunits to one another to produce the oligomers . In the elongation step, two carbohydrate amino acid subunits (CA ' s ) or glycoside amino acid subunits (GA' s ) are coupled to one another via an amide linkage as indicated above .
- the invention is also directed to chemical
- a first chemical intermediate is a derived carbohydrate amino acid having an anomeric carbon and non-anomeric carbons .
- the anomeric carbon is substituted with a carboxyl radical .
- Each of the non-anomeric carbons is substituted with a radical selected from the group consisting of blocked hydroxyl , blocked amino ,
- a second chemical intermediate is a derived carbohydrate amino acid similar to the first except that the non-anomeric carbons are substituted with a radical selected from the group consisting of blocked hydroxyl , blocked amino , unprotected amino , and hydrogen, with the proviso that at least one radical is an
- unprotected amino and at least one radical is a blocked hydroxyl or amino .
- a second aspect of the invention involves the molecular design and chemical synthesis of a class of carbohydrates designated as carbonucleotoids (CND ' s ) .
- Carbonucleotoids are oligosaccharides which employ oligonucleotide-like phosphate bonds for linking the various carbohydrate subunits within an oligomer
- Phosphate bond formation may be achieved by employing technology and instrumentation developed for oligonucleotide synthesis .
- the phosphate bonds employed within carbonucleotoids are convenient linkages for coupling these units . The ease and high efficiency by which the oligonucleotide-like linkages can be
- the disclosed methods are characterized by their versatility and practicality.
- the methods may exploit conventional solid phase and automated synthesis
- oligomeric carbonucleotoid molecule comprising carbohydrate C-glycoside subunits (CG ' s ) coupled to one another via a phosphodiester linkage .
- the phosphodiester linkage may be represented by the structure : CG 1 -C 1 ' - (O-PO (OH) -O) -CG 2 .
- the first carbohydrate C-glycoside subunit (CGi-C 1 ' ) has an anomeric carbon forming a C-glycosidic bond with a carbon C 1 ' . In turn the carbon C 1 ' is bonded to the phosphodiester linkage .
- the second carbohydrate C-glycoside subunit CG2 has a non-anomeric carbon bonded to the phosphodiester linkage .
- the invention is also directed a process for synthesizing the oligomeric carbonucleotoid molecule .
- the process employs a coupling step wherein two or more carbohydrate C-glycoside subunits (CG ' s ) are coupled by means of a phosphodiester linkage as indicated above .
- the second aspect of the invention is also directed to libraries of oligomeric carbonucleotoid molecules .
- the libraries are employable for drug screening .
- Each oligomeric carbonucleotoid molecule including at least two carbohydrate C-glycoside subunits (CG ' s ) coupled to one another by means of a phosphodiester linkage as indicated above .
- the invention is also directed to an improved process for synthesizing a library of oligomers .
- the process employs an elongation step wherein subunits are coupled to one another to produce the oligomers .
- the improvement is directed to the use of phosphodiester linkage linkages for linking the C-glycoside subunits as indicated above .
- the second aspect of the invention is also directed to derived carbohydrate C-glycosides having an anomeric carbon and non-anomeric carbons .
- the anomeric carbon forms a C-glycosidic bond with carbon C 1 ' .
- the carbon C 1 ' is bonded to an phosphoramidite .
- Each of the non-anomeric carbons is substituted with a radical selected from the group consisting of blocked hydroxyl , differentially protected hydroxyl , and hydrogen, with the proviso that at least one radical is a differentially protected hydroxyl .
- An alternative derived carbohydrate C-glycoside is similar to the above except that each of the non-anomeric carbons is substituted with a radical selected from the group consisting of blocked hydroxyl , unprotected hydroxyl , and hydrogen, with the proviso that at least one radical is an unprotected hydroxyl and at least one radical is a blocked hydroxyl .
- the carbopeptoids are oligomers having repeating carbohydrate subunits linked to one another by means of amide linkage units. More particularly, the carbonyl carbon of each amide linkage unit is bonded to the anomeric carbon of a carbohydrate subunit.
- the amide nitrogen of the amide linkage unit is bonded to a non-anomeric carbon.
- the retrosynthetic scheme suggests that the amide bond may be split and that the preferred starting materials are carbohydrate amino acids.
- Carbonucleotoids are oligosaccharides in which carbohydrate C-glycoside subunits (CG's) are linked to one another by means of phosphodiester bonds. More particularly, the retrosynthetic scheme suggests that the phosphate group may be eliminated, yielding hydroxylated starting material.
- Scheme 2 illustrates representative carbohydrate amino acid subunits (CA's) and carbohydrate C-glycoside subunits (CG's).
- Preferred carbohydrate amino acid subunits (CA's) include the following:
- D-glucose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
- D-mannose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
- D-galactose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) pooitions;
- N-acetyl-D-glucosamine having an unprotected carboxyl at the anomeric C(1) position, an
- ⁇ -D-altrose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
- ⁇ -D-gulose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
- ⁇ -D-glucose having an unprotected O-glycosidic amino at the anomeric C(1) position, an unprotected carboxyl as the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
- D-mannose having an unprotected O-glycosidic amino at the anomeric C(1) position, an unprotected carboxyl as the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
- D-galactose having an unprotected O-glycosidic amino at the anomeric C(1) position, an unprotected carboxyl as the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
- N-acetyl-D-glucosamine having an unprotected 0- glycosidic amino at the anomeric C(1) position, an unprotected carboxyl as the C(6) position, a blocked amino group at the C(2) position and blocked hydroxyls at the C(3) and C(4) positions;
- D-ribose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(5) position, and blocked hydroxyls at the C(2) and C(3) positions;
- D-arabinose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(5) position, and blocked hydroxyls at the C(2) and C(3) positions.
- Preferred carbohydrate amino acid subunits include the following:
- D-glucose having a C(1) C 1 -glycosidic carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position and blocked hydroxyls at the C(2), C(3), and C(4) positions;
- D-mannose having a C(1) C 1 -glycosidic carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position and blocked hydroxyls at the C(2), C(3), and C(4) positions;
- D-galactose having a C(1) C 1 -glycosidic carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position and blocked hydroxyls at the C(2), C(3), and C(4) positions;
- N-acetyl-D-glucosamine having a C(1) C 1 -glycosidic carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position, a blocked amino at the C(2) position, and blocked hydroxyls at the C(3) and C(4) positions.
- CA's protected carbohydrate amino acid subunits from N- acetyl-D-glucosamine, i.e. compound 62.
- Scheme 5 summarizes the synthesis of hexamer 74, i.e glucose-glucosamine hetero carbopeptoid (CPD) .
- Scheme 6 illustrates the construction of suitably protected and activated C-glycoside subunits (CG's) corresponding to glucose.
- Scheme 7 illustrates the construction of suitably protected and activated C-glycoside subunits (CG's) corresponding to glucosamine.
- Scheme 8 summarizes the synthesis of hexamer 116, i.e. glucose-glucosamine hetero carbonucleotoid (CND) .
- a oligosaccharide carbopeptoid (CPD) library may be constructed by performing using a split synthesis method of oligomerization as illustrated in Scheme 500 for carbopeptoids and Scheme 550 for carbonucleotoids .
- the split synthesis may employ beads upon which to build the oligomers . Beads are aliquoted into each of a several reaction vessels , each reacrtion vessel containing a different core molecule . The core molecules are then allowed to attach to the beads .
- the beads are washed, mixed with one another , and then re-aliquoted ( split ) into a second set of reaction vessels for addition of a second core molecule to the first added core molecule .
- the process is then reiterated until the oligomerization process is complete .
- the resultant library of oligosaccharides may then be screened using conventional methods developed for oligopeptide and oligonucleotide libraries . Screening an oligosaccharide library can lead to the identification of individual oligosaccharide components within the library having binding activity and/ or bioactivity.
- oligosaccharide libraries may be enlarged by introducing additional functionalities into the basic CA ' s and CG ' s .
- oligosaccharide libraries may be further enlarged by enlarging the pool of free functional groups on the CA ' s and CG ' s and employed this enlarged pools of CA ' s and CG ' s during the respective split synthesis processes .
- Scheme 20 illustrate a protocol published by Fuchs, E.F. et al. (J. Chem Ber. 1975, 108, 2254) for the synthesis of CA 45 and 46 from glucose pentaacetate.
- Scheme 20 illustrates a synthetic route for CG 82, also starting from glucose pentaacetate.
- the reagents and conditions for synthesizing CG 82 are provided as follows:
- Step (e) (1) DMTC1, DMAP, Pyridine; room temperature .
- Step (f ) DIBAL-H, CH 2 C12 ; -78°C;
- Step (g) (NCCH 2 CH 2 ) (NiPr2)PCl, tetrazole, CH 2 Cl 2 .
- Step M The reagents and conditions for synthesizing CA 46 from CA 45 are provided in Step M as follows:
- Step (m) FMOC-Cl, K 2 CO 3 , THF, H 2 O; 0°C .
- Step (b) (1) AcOH, H 2 0, THF;
- Step (d) (1) DMTCl, DMAP, Pyridine, room temperature;
- Step (e) BH3-THF
- Step (f) (NCCH 2 CH 2 ) (NiPr 2 )PCl, tetrazole, Ch 2 Cl 2 ;
- Step (g) (1) 1 equiv TsCl. base;
- Step (h) NaN 3 ;
- Step (i) H 2 , Pd(OH) 2 -C;
- Step (j) FMOC-Cl, base.
- Step (a) reductive debenzylation
- Step (b) (1) equiv TsCl. base;
- Step (c) NaN 3 .
- Step (d) RuCl 3 , NalO 4 , CH 3 CN, H 2 O, CCl 4 .
- Step (e) H 2 , Pd-C.
- Step (f) FMOC-Cl, base.
- Step (g) (1) DMTCl, DMAP, Pyridine, room
- Step (h) (1) RuCl 3 , NalO 4 , CH 3 CN, H 2 O, CCl 4 ;
- Step (j) PPh3 , DIAD, diphenyl phosphoryl azide (DPPA) ,THF.
- Step (k) KMnO4, t-BuOH, buffer.
- Step (a) (1) 1.1 equivalent DMTCl, DMAP, Pyridine,
- Step (b) (1) LAH, ether, reflux, 2 hour;
- Step (c) 10% HCOOH in CH 2 Cl 2 , 0°C, 2 minutes, 100%.
- Step (d) RuCl3, NalO 4 , CH 3 CN, H 2 O, CCl 4 , 20°C, 10 minutes,
- Step (e) (1) 1 equiv. TsCl, base;
- Step (f) NaN 3 .
- Step (g) oxidative NEF.
- Step (h) Pd-C, H 2 .
- Step (i) FMOC-Cl, base.
- Step (j) (1) 1 equiv. PivCl, base;
- Step (k) (1) oxidative Nef; (2) CH 2 N 2 .
- Step (m) DMTCl, DMAP, Pyridine.
- Step (n) LAH.
- Step (p) LAH.
- Step 1 DCC, HOBT, Et 3 , DMF;
- the crude product 50 is next dissolved in ethanol (0.15 M) and then concentrated H2S O 4 (0.01 equivalents-catalytic) is added. The reaction mixture is heated to 85 °C for eight hours.
- the crude product 126 is dissolved in 25% NaOH (0.5 M) and heated at reflux for 18 hours (vigorous reflux is necessary). Next, the solution is diluted with an addition of water (0.1 M) and to this solution is added Amberlite 112120 resin (H + -form) and is then stirred. The supernatant is then decanted and the resin is washed until the eluate is colorless. The eluate is then collected, condensed and azeotroped with MeOH which yields 127 as a crude, pale yellow syrup (47%).
- triol 178 (.0 equiv.) in CH 2 CI 2 (.5 M) at 0 °C, was added triethylamine ( 1.2 equiv.), 4-DMAP (.10 equiv.) and then TOSCl (1.1 equiv.). The reaction is stirred for 1 h and then is quenched with saturated ammonium chloride ( 1.5 mL), diluted with ethyl acetate (25 mL), washed with water (2X 5 mL), brine (1X 5 mL), back-extracted (2X), recombined, dried (MgSO 4 ) and evaporated.
- triol 182 (.0 equiv.) in CH2CI2 (.5 M) at 0 °C
- sodium-azide (1.2 equiv.) from Aldrich chemical company at 0 °C.
- the reaction is stirred for 1 h and then is quenched with saturated ammonium chloride ( 1.5 mL), diluted with ethyl acetate (25 mL), washed with water (2X 5 mL), brine (1X 5 mL), back-extracted (2X), recombined, dried (MgSO 4 ) and evaporated.
- the compound is purified by flash column chromatography and affords compound 183.
- a solution of 201 (1.0 equivalents) is dissolved in ethanol (.01 M total) at 25 °C.
- the mixture is next exposed to 10% Pd/C (.1 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atmosphere.
- the reaction is stirred for 72 hours and is then filtered through celite.
- the crude mixture is subsequently diluted with ether and washed with NaHCO 3 (3X), brine (1X) and dried (MgSO 4 ) and concentrated.
- LAH lithiumaluminumhydride
- M is added lithiumaluminumhydride (LAH) ( 1.5 equivalents) at 30 °C.
- LAH lithiumaluminumhydride
- the reaction is refluxed for 2 hours and then quenched with methanol and diluted with ether.
- the reaction is next worked-up with sodium potassium tartrate (2X), brine (1X) and is then dried (MgSO 4 ) and concentrated.
- the crude mixture is resuspended in methylene chloride (.10 Molar) and to it is added sodium bicarbonate (2.0 equivalents) at 0 °C.
- DMT chloride dimethyoxytritylchloride
- 2X ammonium chloride
- copper sulfate (2X) copper sulfate
- brine (1X) dried over MgSO 4 and concentrated.
- a solution of the crude intermediate 1.0 equivalents is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 °C.
- a depiction of the generation of a combinatorial library for oligopeptoid compounds is shown in scheme 500.
- the example uses an alphabet of eight D-aldose hexose sugars (other sugars groups such as the D/L ketoses and L-configurations of aldose hexoses, may be used) and carries the synthesis to a degree of three or 512 compounds. (The process can repeat itself to afford the library of desired size). Standard chemistry is shown and follows the reaction conditions as described above herein for peptoid synthesis.
- the solid support used is the standard N-(2- Aminoethyl)-3-amino-propyl glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support
- a depiction of the generation of a combinatorial library for oligonucleotoid compounds is shown in scheme 550.
- the example uses an alphabet of eight D-aldose hexose sugars (other sugars groups such as the D/L ketoses and L-configurations of aldose hexoses, may be used) and carries the synthesis to a degree of three or 512 compounds. (The process can repeat itself to afford the library of desired size). Standard chemistry is shown and follows the reaction conditions as described above herein for carbonucleotoid synthesis.
- the solid support used is the standard N-(2-Aminoethyl)-3-amino-propyl glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
- a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
- the benzylidene is then azeotroped with benzene (2X 100 mL) and then dried overnight under vacuum over P 2 O 5 .
- a mixture of benzylidene, dibutyl tin oxide ( 1.2 equiv.) and dry methanol (.25 M) are heated at reflux for 4 h until the solution became clear and homogeneous. (An automatic stirring apparatus may be necessary.)
- the solvent is next removed in vacuo to give a foamy white tin complex which was then azeotroped with benzene (2X) and dried (2 h to overnight) under vacuum over P 2 O 5 .
- the solid support used is the standard N-(2- Amino ethyl)- 3 -amino -propyl glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
- a linker extending from the amino group on the support eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.
- TBDPS ether is then azeotroped with benzene (2X 100 mL) and then dried overnight under vacuum over P 2 O 5 .
- the solid support used is the standard N-(2- Aminoeteyl)-3-amino-propyI glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
- a linker extending from the amino group on the support eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.
- Aqueous layer is back extracted with ethyl acetate (3X) and then recombined with the organic layer which was then dried over MgSO 4 and evaporated. Purification by flash column chromatography yields the desired benzyl ether 2130.
- the compound 2140 is then treated with tetrabutylammonium fluoride (2.0 equivalents) in THF (.1 Molar) and allowed to stir for an additional 2 hours at 25 °C.
- a saturated solution of ammonium chloride (50 mL) is then added dropwise to quench the reaction mixture at 0 °C and the mixture was diluted with ethyl acetate, washed with water (2X), brine (1X), dried overMgSO 4 and evaporated. Purification by flash column chromatography yields tribenzyl ether 2150.
- N- (2-Aminoethyl)-3-amino-propyl glass support amino- polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
- a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
- the concentrate is allowed to cool to room temperature and the product crystallizes overnight and carried on as follows:
- the methyl glycoside is dissolved in chloroform (.5 M) and to it, is added phthalic anhydride ( 1.5 equiv.) and the reaction mixture is allowed to reflux at 70 °C for 4 h.
- phthalic anhydride 1.5 equiv.
- Phosphoramidate 138 (2 diastereomers) : IR, (neat) cm -1 : 3089, 2964, 2927, 2856, 2253, 1497, 1455, 1396, 1363, 1253, 1184, 1156, 1094, 1028, 978, 876, 836, 779, 735,
Abstract
Librairies are synthesized with oligomeric carbopeptoids and carbonucleotoids. Carbopeptoids are oligosaccharides having carbohydrate subunits linked to one another by amide bonds. Carbonucleotoids are oligosaccharides having carbohydrate subunits linked to one another by phosphodiester bonds. Carbopeptoid librairies may be fabricated using automated polypeptide synthesizers. Carbonucleotoid librairies may be fabricated using automated polynucleotide synthesizers.
Description
CARBOPEPTOIDS AND CARBONUCLEOTOIDS Specification Field of the Invention:
The invention relates to oligosaccharides and libraries incorporating oligosaccharide. More
particularly, the invention relates to oligosaccharides and libraries of oligosaccharides which employ amide and/or phosphodiester linkages for joining adjacent carbohydrate subunits.
Background:
Carbohydrates are known to mediate many cellular recognition processes. Carbohydrates can serve directly as binding molecules and, in such instances, are
essential to the recognition process. A review of the biological role of carbohydrates with respect to cellular recognition phenomena is provided by Sharon et al.
(Scientific American, January 1993, 82). The emerging importance of glycobiology is further characterized by Mekelburger et al. (Angew. Chem. Int. Ed. Engl. 1992, 31, 1571) and by Dagani et al. (Chem. Eng. News, February 1, 1993, 28).
Dysfunctional mediation of cellular recognition processes can lead to disease states. If a cellular recognition process is mediated by an oligosaccharide, then an absence or excess of such oligosaccharide can lead to a dysfunctional mediation of such process. The
mediating oligosaccharide may be deficient or absent due to a deficiency of production or due to a high rate of catabolism. If rate of catabolism is excessive, then catabolically resistant analogs of the bioactive
oligosaccharide may be preferred as drug candidates as compared to the native bioactive oligosaccharide .
Accordingly, what is needed is a library which includes analogs of known bioactive oligosaccharides . Such a library may be usefully employed for screening drug candidates .
Central requirements for the design of libraries of oligosaccharide analogs include the following :
(a) A need to maximize the potential of the designed oligosaccharides as ligand and drug candidates ;
(b) A need to capitalize on existing highly sophisticated technology directed to the synthesis of oligopeptides and oligonucleotides in order to facilitate the rapid and efficient design and construction of oligosaccharides ; and
(c ) A need for flexibility with respect to
synthesizing either single target molecules or large libraries of target molecules simultaneously .
Methodologies for synthesizing biopolymers are well developed for peptides , nucleic acids , and saccharides .
Segments of oligopeptides and of oligonucleotides can now be routinely synthesized both in solution and in the solid phase , manually and/or on automated systems . The synthesis of such structures is facilitated by the
availability of efficient techniques and sophisticated instrumentation for synthesizing peptide and phosphate bonds with high yields . The synthesis of oligopeptides and oligonucleotides is also facilitated by the absence of stereocenters in these linkages . In contrast , technology for the construction of oligosaccharides is comparatively less sophisticated and efficient .
Synthetic methods for constructing oligosaccharides give comparatively lower yields and are complicated by the two isomer possibilities (α and β) in glycoside bond
formation .
Techniques and chemical methods for simultaneously synthesizing multiple oligopeptides , e . g . 100-150
completely different peptides having lengths of up to 20 amino acid residues , are reviewed by Jung, G . et al .
(Angew. Chem, Int . Ed. Engl . 1992 , 31 , 367-383 - incorporated therein by reference) . Such techniques facilitate the construction of oligopeptide libraries .
Simon, et al . ( Proc, Natl . Acad. Sci . USA, 1992 , 89, 9367-9371 ) disclose oligopeptide analogs in which amino acid side chain groups are attached not to conventional peptide backbone carbons but to peptide backbone
nitrogens . Such analogs are termed peptoids . Simon also discloses the construction of peptoid libraries as a modular approach to drug discovery . Simon ' s
oligopeptoids are shown by calculation to have greater conformational freedom as compared to conventional oligopeptides . Accordingly, oligopeptoids are thought to have greater potential as pharmaceutically useful binding
ligands as compared to conventional oligopeptides having close sequence homology to such oligopeptoids .
Von Roedern et al . disclose a carbohydrate amino acid {Angew. Chem, Int . Ed. Engl . 1994 , 31 , 687-689 ) . Although von Roedern discloses that carbohydrate amino acids may be coupled to peptides , he does not disclose that they may also be polymerized so as to form
oligosaccharides .
Summary :
A first aspect of the invention involves the molecular design and chemical synthesis of a class of carbohydrates designated as carbopeptoids (CPD ' s ) .
Glycopeptoids are preferred carbopeptoids . Carbopeptoids and glcopeptoids are oligosaccharides which employ peptide- like amide bonds for linking the various
carbohydrate subunits within an oligomer assembly. Amide bond formation may be achieved by employing oligopeptide synthesis technology and instrumentation . The method allows for the design and synthesis of specific compounds for biological and pharmacological investigations . The method also allows for the generation of libraries of compounds for biological and pharmacological screening . Conventional screening techniques employed with respect to peptide and peptoid libraries (Simon et al . , supra) may also be employed with respect to carbopeptoid libraries . The design takes advantage of the
multifunctionality of carbohydrate subunits to maximize
the binding properties of the molecules . The ease and high efficiency by which the peptide-like linkages can be constructed make the synthesis of these molecules a practical proposition . Furthermore , non-carbohydrate units may be inserted into the sequence making this approach even more flexible and versatile for the generation of new libraries of organic compounds .
More particularly, the invention is directed to a oligomeric carbopeptoid or glycopeptoid compound having carbohydrate amino acid subunits (CA ' s ) or glycoside amino acid subunits (GA' s ) coupled to one another via an amide linkage . The amide linkage may be represented by the formula CA1 - (CO-NH) -CA2 . The amide linkage (CO-NH) includes a carbonyl carbon and an amido nitrogen . A first carbohydrate amino acid subunit CA1 or glycoside amino acid subunit GA1 has an anomeric carbon bonded to the carbonyl carbon of the amide linkage . The anomeric carbon of the first carbohydrate amino acid subunit CA1 forms a C-glycosidic bond with the carbonyl carbon of the amide linkage and maintains the carbohydrate in a closed ring configuration . A second carbohydrate amino acid subunit CA2 has a non-anomeric carbon bonded to the amido nitrogen of the amide linkage . The second carbohydrate amino acid subunit CA2 , like the first amino acid subunit CA1 ( may include an anomeric carbon bonded to the
carbonyl carbon of a second amide linkage linking the second carbohydrate amino acid subunit CA2 to a third carbohydrate amino acid subunit CA3 , etc . In this instance , the anomeric carbon of the second
carbohydrate amino acid subunit CA2 forms a C-glycosidic bond with the carbonyl carbon of the amide linkage and maintains the carbohydrate in a closed ring
configuration . On the other hand, if the second
carbohydrate amino acid subunit CA2 is a terminal subunit , then its anomeric carbon may form a hemiacetal , a hemiketal , or a glycoside .
The invention is also directed to a process for synthesizing the above oligomeric carbopeptoid or glycopeptoid compound. The synthetic process involves the coupling of two or more carbohydrate amino acid subunits (CA ' s ) or glycoside amino acid subunits (GA' s ) to one another by means of amide linkages .
The invention is also directed to libraries of oligomeric carbopeptoid or glycopeptoid compounds . Such libraries are employable for drug screening . Each oligomeric carbopeptoid or glydopeptoid compound includes at least two carbohydrate amino acid subunits (CA ' s ) or glycoside amino acid subunits (GA' s ) coupled to one another via an amide linkage as indicated above . The invention is also directed to an improved process for synthesizing the above library of oligomers . The process employs an elongation step for coupling the subunits to one another to produce the oligomers . In the elongation step, two carbohydrate amino acid subunits (CA ' s ) or glycoside amino acid subunits (GA' s ) are coupled to one another via an amide linkage as indicated above .
The invention is also directed to chemical
intermediates for producing oligomeric carbopeptoids . A
first chemical intermediate is a derived carbohydrate amino acid having an anomeric carbon and non-anomeric carbons . The anomeric carbon is substituted with a carboxyl radical . Each of the non-anomeric carbons is substituted with a radical selected from the group consisting of blocked hydroxyl , blocked amino ,
differentially protected amino , and hydrogen, with the proviso that at least one radical is a differentially protected amino . A second chemical intermediate is a derived carbohydrate amino acid similar to the first except that the non-anomeric carbons are substituted with a radical selected from the group consisting of blocked hydroxyl , blocked amino , unprotected amino , and hydrogen, with the proviso that at least one radical is an
unprotected amino and at least one radical is a blocked hydroxyl or amino .
A second aspect of the invention involves the molecular design and chemical synthesis of a class of carbohydrates designated as carbonucleotoids (CND ' s ) . Carbonucleotoids are oligosaccharides which employ oligonucleotide-like phosphate bonds for linking the various carbohydrate subunits within an oligomer
assembly . Phosphate bond formation may be achieved by employing technology and instrumentation developed for oligonucleotide synthesis . The phosphate bonds employed within carbonucleotoids are convenient linkages for coupling these units . The ease and high efficiency by which the oligonucleotide-like linkages can be
constructed make the synthesis of these molecules a
practical proposition .
The disclosed methods are characterized by their versatility and practicality. The methods may exploit conventional solid phase and automated synthesis
techniques for producing carbopeptoids and
carbonucleotoids in large scale .
More particularly, the second aspect of the
invention is directed to an oligomeric carbonucleotoid molecule comprising carbohydrate C-glycoside subunits (CG ' s ) coupled to one another via a phosphodiester linkage . The phosphodiester linkage may be represented by the structure : CG1-C1 ' - (O-PO (OH) -O) -CG2. The first carbohydrate C-glycoside subunit (CGi-C1 ' ) has an anomeric carbon forming a C-glycosidic bond with a carbon C1 ' . In turn the carbon C1 ' is bonded to the phosphodiester linkage . The second carbohydrate C-glycoside subunit CG2 has a non-anomeric carbon bonded to the phosphodiester linkage . The invention is also directed a process for synthesizing the oligomeric carbonucleotoid molecule . The process employs a coupling step wherein two or more carbohydrate C-glycoside subunits (CG ' s ) are coupled by means of a phosphodiester linkage as indicated above .
The second aspect of the invention is also directed to libraries of oligomeric carbonucleotoid molecules . The libraries are employable for drug screening . Each oligomeric carbonucleotoid molecule including at least two carbohydrate C-glycoside subunits (CG ' s ) coupled to one another by means of a phosphodiester linkage as indicated above . The invention is also directed to an
improved process for synthesizing a library of oligomers . The process employs an elongation step wherein subunits are coupled to one another to produce the oligomers . The improvement is directed to the use of phosphodiester linkage linkages for linking the C-glycoside subunits as indicated above .
The second aspect of the invention is also directed to derived carbohydrate C-glycosides having an anomeric carbon and non-anomeric carbons . The anomeric carbon forms a C-glycosidic bond with carbon C1 ' . In turn, the carbon C1 ' is bonded to an phosphoramidite . Each of the non-anomeric carbons is substituted with a radical selected from the group consisting of blocked hydroxyl , differentially protected hydroxyl , and hydrogen, with the proviso that at least one radical is a differentially protected hydroxyl . An alternative derived carbohydrate C-glycoside is similar to the above except that each of the non-anomeric carbons is substituted with a radical selected from the group consisting of blocked hydroxyl , unprotected hydroxyl , and hydrogen, with the proviso that at least one radical is an unprotected hydroxyl and at least one radical is a blocked hydroxyl .
Detailed Description:
Retrosynthetic schemes for carbopeptoids (compound I) and carbonucleotoids (compound II) are illustrated in Scheme 1.
The carbopeptoids (CPD's) are oligomers having repeating carbohydrate subunits linked to one another by means of amide linkage units. More particularly, the carbonyl carbon of each amide linkage unit is bonded to the anomeric carbon of a carbohydrate subunit.
Similarly, the amide nitrogen of the amide linkage unit is bonded to a non-anomeric carbon. The retrosynthetic scheme suggests that the amide bond may be split and that the preferred starting materials are carbohydrate amino acids.
Carbonucleotoids (CND's) are oligosaccharides in which carbohydrate C-glycoside subunits (CG's) are linked to one another by means of phosphodiester bonds. More particularly, the retrosynthetic scheme suggests that the phosphate group may be eliminated, yielding hydroxylated starting material.
Scheme 2 illustrates representative carbohydrate amino acid subunits (CA's) and carbohydrate C-glycoside subunits (CG's). Preferred carbohydrate amino acid subunits (CA's) include the following:
D-glucose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
D-mannose having an unprotected carboxyl at the
anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
D-galactose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) pooitions;
unprotected amino group at the C(6) position, a blocked amino group at the C(2) position, and blocked hydroxyls at the C(3) and C(4) positions; α-D-idose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
α-D-altrose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
α-D-gulose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
α-D-glucose having an unprotected O-glycosidic amino at the anomeric C(1) position, an unprotected carboxyl as the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
D-mannose having an unprotected O-glycosidic amino at the anomeric C(1) position, an unprotected carboxyl as the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
D-galactose having an unprotected O-glycosidic amino at the anomeric C(1) position, an unprotected carboxyl as the C(6) position, and blocked hydroxyls at the C(2), C(3), and C(4) positions;
N-acetyl-D-glucosamine having an unprotected 0- glycosidic amino at the anomeric C(1) position, an unprotected carboxyl as the C(6) position, a blocked amino group at the C(2) position and blocked hydroxyls at the C(3) and C(4) positions;
D-ribose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(5) position, and blocked hydroxyls at the C(2) and C(3) positions; and
D-arabinose having an unprotected carboxyl at the anomeric C(1) position, an unprotected amino group at the C(5) position, and blocked hydroxyls at the C(2) and C(3) positions. Preferred carbohydrate amino acid subunits (CA's) include the following:
D-glucose having a C(1) C1 -glycosidic carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position and blocked hydroxyls at the C(2), C(3), and C(4) positions;
D-mannose having a C(1) C1 -glycosidic carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position and blocked hydroxyls at the C(2), C(3), and C(4) positions;
D-galactose having a C(1) C1 -glycosidic carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position and blocked hydroxyls at the C(2), C(3), and C(4) positions; and
N-acetyl-D-glucosamine having a C(1) C1 -glycosidic
carbon bonded to a phosphoramidite, an unprotected hydroxyl at the C(6) position, a blocked amino at the C(2) position, and blocked hydroxyls at the C(3) and C(4) positions.
Scheme 3 outlines a preferred synthesis of suitably protected carbohydrate amino acid subunits (CA's) from D- glucose, i.e. compound 46.
protected carbohydrate amino acid subunits (CA's) from N- acetyl-D-glucosamine, i.e. compound 62.
Scheme 5 summarizes the synthesis of hexamer 74, i.e glucose-glucosamine hetero carbopeptoid (CPD) .
Scheme 6 illustrates the construction of suitably protected and activated C-glycoside subunits (CG's) corresponding to glucose.
Scheme 7 illustrates the construction of suitably protected and activated C-glycoside subunits (CG's) corresponding to glucosamine.
Scheme 8 summarizes the synthesis of hexamer 116, i.e. glucose-glucosamine hetero carbonucleotoid (CND) .
The chemistries illustrated in Schemes 5 and 8 for synthesizing heterohexamer CPD 74 and heterohexamer CND 116 can also be employed for synthesizing homohexamer CPD ' s 118 (glucose) and 120 (glucosamine) and homohexamer CND ' s 122 (glucose) and 124 (glucosamine) .
In analogy with the construction of oligopeptide and oligonucleotide libraries , a oligosaccharide carbopeptoid (CPD) library may be constructed by performing using a split synthesis method of oligomerization as illustrated in Scheme 500 for carbopeptoids and Scheme 550 for carbonucleotoids . For example , the split synthesis may employ beads upon which to build the oligomers . Beads are aliquoted into each of a several reaction vessels , each reacrtion vessel containing a different core molecule . The core molecules are then allowed to attach to the beads . The beads are washed, mixed with one another , and then re-aliquoted ( split ) into a second set of reaction vessels for addition of a second core molecule to the first added core molecule . The process is then reiterated until the oligomerization process is complete . The resultant library of oligosaccharides may then be screened using conventional methods developed for oligopeptide and oligonucleotide libraries . Screening an oligosaccharide library can lead to the identification of individual oligosaccharide components within the library having binding activity and/ or bioactivity.
The above oligosaccharide libraries (CPD and CND) may be enlarged by introducing additional functionalities into the basic CA ' s and CG ' s .
The above oligosaccharide libraries (CPD and CND) may be further enlarged by enlarging the pool of free functional groups on the CA ' s and CG ' s and employed this enlarged pools of CA ' s and CG ' s during the respective split synthesis processes .
Scheme 20 illustrate a protocol published by Fuchs, E.F. et al. (J. Chem Ber. 1975, 108, 2254) for the synthesis of CA 45 and 46 from glucose pentaacetate.
Additionally, Scheme 20 illustrates a synthetic route for CG 82, also starting from glucose pentaacetate. The reagents and conditions for synthesizing CG 82 are provided as follows:
Steps (a) -(d): according to Fuchs (supra). Step (e) : (1) DMTC1, DMAP, Pyridine; room temperature .
(2) TESTfl; 0°C.
Step (f ) : DIBAL-H, CH2C12 ; -78°C; and
Step (g) : (NCCH2CH2) (NiPr2)PCl, tetrazole, CH2Cl2.
The reagents and conditions for synthesizing CA 46 from CA 45 are provided in Step M as follows:
Step (m) : FMOC-Cl, K2CO3 , THF, H2O; 0°C .
Scheme 20. Conventional route to CAs and a variation for the synthesis of CG. Reagents and conditions: (a) - (d) corresp. Lit.: (e) (1) DMTCl, DMAP, Py; RT. (2) TESTfl; 0 °C. (f) DIBAL-H, CH2Cl2; -78 °C. (g)
(NCCH2 CH2)(NiPr2)PCl, tetrazole CH2Cl2. (m) FMOC-Cl, K2CO3, THF, H2O; 0 ºC.
A synthetic route for producing C-glycosides (CG's) with β-conf iguration at the former anomer center is illustrated in Scheme 21. The starting material
(compound 36) is commercially available. The reagents and conditions for synthesizing CG 181 and CG 185 are as follows :
Step (a): Co2(CO)8, HSiEt2Me, CO.
Step (b) : (1) AcOH, H20, THF;
(2) RuCl3, NalO4, CH3CN, H2O, CCL4, room temperature;
Step (c) : NaOMe, MeOH;
Step (d) : (1) DMTCl, DMAP, Pyridine, room temperature;
(2) TESOTf ;
Step (e) : BH3-THF;
Step (f) : (NCCH2CH2) (NiPr2)PCl, tetrazole, Ch2Cl2 ;
Step (g) : (1) 1 equiv TsCl. base;
( 2 ) TESOTf ;
Step (h) : NaN3;
Step (i) : H2, Pd(OH)2-C;
Step (j) : FMOC-Cl, base.
Scheme 21. Synthesis of C-glycosides with β-conf iguration at the former anomeric center. Reagents and conditions: (a) Co2(CO)8, HSiEt2Me, CO. (b) (1) AcOH, H2O, THF; (2) RuCl3, NaIO4, CH3CN, H2O, CCl4, RT. (c) NaOMe, MeOH. (d) (1) DMTCl, DMAP, Py, RT: (2) TESOTF. (e) BH3-THF. (f)
(NCCH2CH3)(NiPr2)PCl, tetrazole, CH2Cl2. (g) (1) 1 equiv TsCl, base: (2) TESOTf (H) NaN3."(i) H2, Pd(OH)2-C. (j) FMOC-Cl, base.
Scheme 22. Synthesis of C-glycosides with α-configuration at the former anomeric center. Reagents and conditions: (a) reductive debenzylation. (b) (1) 1 equiv TsCl, base; (2) TESOTf. (c) NaN3. (d) RuCl3, NaIo4, CH3CN, H2O, CCl4. (e) H2, Pd-C. (f) FMOC-Cl, base, (g) (1) DMTCl, DMAP, Py, RT; (2) TESOTf. (h) (1) RuCl3, NaIO4, CH3CN, H2O, CCl4; (2) CH2N2. (i) DIBAL-H. (j) PPh3, DIAD, diphenyl phosphoryl azide (DPPA), THF. (k) KMnO4, t-BuOH, buffer.
Synthetic routes for producing with C-glycosides with α-conf igurations at the former anomeric center, i.e. CG 196 and CG 1204, are illustrated in Scheme 22. The common starting material for these synthetic routes
(compound 190) is disclosed by Schmidt, R. R. et al.
(Liebigs Ann. Chem. 1987, 825). The reagents and
conditions for the reactions leading to CG 196 and CG 204 are as follows :
Step (a) : reductive debenzylation;
Step (b) : (1) equiv TsCl. base;
(2) TESOTf.
Step (c) : NaN3.
Step (d) : RuCl3, NalO4, CH3CN, H2O, CCl4.
Step (e) : H2, Pd-C.
Step (f) : FMOC-Cl, base.
Step (g) : (1) DMTCl, DMAP, Pyridine, room
temperature;
(2) TESOTf.
Step (h) (1) RuCl3, NalO4, CH3CN, H2O, CCl4;
(2) CH2N2.
Step (i) DIBAL-H.
Step (j) PPh3 , DIAD, diphenyl phosphoryl azide (DPPA) ,THF.
Step (k) KMnO4, t-BuOH, buffer.
Reactions for the development of the galactose derived C-glycoside 138 into protected CA's and diols is illustrate in Scheme 23. The common starting material for these synthetic routes (compound 138) is disclosed by
Petrus, L. et al. (Chem. zvesti. 1982, 36, 103). The reagents and conditions required for the synthesis of compound 209, compound 214, compound 220, and compound 224 are indicated below:
Step (a) : (1) 1.1 equivalent DMTCl, DMAP, Pyridine,
12 hour, 20°C;
(2) TesOTf, CH2, 0°C, 1 hour, 83%.
Step (b) : (1) LAH, ether, reflux, 2 hour;
(2) FMOC-Cl, K2CO3, THF, H2O, 0°C, 1 hour, 55%;
Step (c) : 10% HCOOH in CH2Cl2, 0°C, 2 minutes, 100%. Step (d) : RuCl3, NalO4, CH3CN, H2O, CCl4, 20°C, 10 minutes,
54%.
Step (e) : (1) 1 equiv. TsCl, base;
(2) TESOTf.
Step (f) : NaN3.
Step (g) : oxidative NEF.
Step (h) : Pd-C, H2.
Step (i) : FMOC-Cl, base.
Step (j) : (1) 1 equiv. PivCl, base;
(2) TESOTf.
Step (k) : (1) oxidative Nef; (2) CH2N2.
Step (l) : DIBAL-H.
Step (m) : DMTCl, DMAP, Pyridine.
Step (n) : LAH.
Step (o) : Nef reaction
Step (p) : LAH.
Scheme 23. Development of the galactose derived C-glycoside 138 to
protected CAs and diols. Reagents and conditions: (a) (1) 1.1 equiv DMTCl, DMAP, Py, 12h, 20 °C; (2) TesOTf, CH2C12, 0 °C, lh, 83%. (b) (1) LAH, ether, reflux, 2h; (2) FMOC-Cl, K2CO3, THF, H2O, 0 °C, lh, 55%. (c) 10% HCOOH in CH2Cl2 0 °C, 2 min, 100%. (d) RuCl3, NaIO4, CH3CN, H2O, CCl4, 20 °C, 10 min, 54%. (e) (1) 1 equiv TsCl, base; (2) TESOTf. (f) NaN3. (g) oxidative Nef. (h) Pd-C, H2. (i) FMOC-Cl, base, (j) (1) 1 equiv PivCl, base; (2) TESOTf. (k) (1) oxidative Nef; (2) CH2N2. (1) DIBAL-H. (m) DMTCl, DMAP, Py. (n) LAH. (o) Nef reaction, (p) LAH
An exemplary protocol for synthesizing a hexamer carbopeptoid (CPD 234) starting from galactose derived CA 214, glucosamine derived CA 62, and glucose derived CA, using standard methods for solid phase peptide synthesis is illustrated in Scheme 24. The reagents and condition for these reactions are as follows:
Step 1: DCC, HOBT, Et3 , DMF;
Step 2: Piperidine, DMF
SYNTHETIC METHODS
Preparation of 37
To a solution of β -D-Glucose pentaacetate 36 i n nitromethane from Aldrich company (.13 Molar), is added trimethylsilyl cyanide (3.0 equivalents) and then SnCl4 ( .02 equivalents). The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the remaining trimethylsilyl cyanide. The mixture is evaporated and the remaining oil is resuspended in dichloromethane and washed with sodium acetate solution (1X), water (1X), brine (1X) and then dried over magnesium sulphate and concentrated. The crude solid is then recrystallized from methanol to yield 37 as a white solid (47%). scheme 3 step 1; scheme 9, step a.
Preparation of 38
The crude product 37 is next dissolved in ethanol (0.15 M) and then concentrated H2S O 4 (0.01 equivalents-catalytic) is added. The reaction mixture is heated to 85 °C for eight hours. The solution is next concentrated in vacuo and purification by flash column chromatography affords compound 38. scheme 3 step 2
Preparation of 39
To a solution of 38 ( 1.0 equivalents) in pyridine (.10
Molar), is added trimethylacetyl chloride (pivaloyl chloride) (2.5 equivalents) at 0 °C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium
chloride (2X), copper sulfate (2X), brine (1X), dried over MgSO4 and concentrated. Purification by flash column chromatography affords compound 39. scheme 3 step 1 Preparation of 40
(.10 Molar), is added diisopropylethylamine (3.3 equivalents) at 0 ° C . S ub sequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4 and concentrated. Purification by flash column chromatography affords compound 40. scheme 3 step 2
Preparation of 41
To a solution of 40 in ethanol (.13 Molar), is added sodium ethoxide (0.3 equivalents) and the reaction mixture is stirred for two hours at room temperature. The solution is then concentrated in vacuo and purification by flash column chromatography affords compound 41. scheme 3 step 1
Preparation of 42
A solution of 41 ( 1.0 equivalents) in tetrahydrofuran (.18
M) is treated with DPPA (diphenylphosphorylazide, 2.0 equivalents), triphenylphosphine ( 1.3 equivalents) and DIAD (diisopropyl-azo-dicarboxylate, 1.3 equivalents). The reaction is heated to 80 °C for 3 hours and then diluted with ether (2X) and
washed with .5 M aqueous NaOH (2X). The organic layer is dried over MgSO4 and evaporated. Purification by flash column chromatography affords compound 42. scheme 3 step 2
Preparation of 44
(.01 M total) at 25 °C. The mixture is next exposed to 10% Pd/C (.1 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atmosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 44. scheme 3 step 1
Preparation of 45
A solution of 44 (1.0 equivalents) is dissolved in p- dioxanes (.1 M) and then exposed to a solution 3.0 Molar solution of sodium hydroxide ( 1.5 equivalents). The reaction is then stirred for 2 hours at 50 °C and is subsequently diluted with ether and washed with a solution of NH4CI (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 45. scheme 3 step 1
Preparation of 46
To a solution of 45 ( 1.0 equivalents) in methylene chloride (.10 Molar), is added sodium bicarbonate (2.0 equivalents) at 0 °C. Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-Cl, 1.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried ( M g S O 4 ) and concentrated. Purification by flash column
chromatography affords compound 46. scheme 3 step 2
Preparation of 48
Procedure as described in Methods in Carbohydrate chemistry, Whistler, R., II , 1963 , p. 327. A mixture of 80g anhydrous D-glucosamine hydrochloride or D-galactosamine hydrochloride from Aldrich chemical company, in 200 mL. methanol and 20g Dowex 50 (H+) acidic resin, is stirred at the boiling point in a round bottom flask. After 24-hr. reaction time, the resin is removed by filtration and ished three times with 20 ml. of methanol. The filrate and ishings are combined and concentrated to about 125 ml by rotovap. The concentrate is allowed to cool to room temperature and the product crystallizes overnight.
To a solution of free amine, in chloroform (.5 M), is added phthalic anhydride ( 1.5 equiv.) and the reaction mixture is allowed to reflux at 70 °C for 4 h. The product is then crystallized and carried onto the next step.
To a solution of triol in methylene chloride (.5 M), is added
acetic anhydride (3.5 equiv.) and triethyl amine (3.5 equiv.) and the reaction mixture is allowed to stir at 0 °C for 4 h. The product 48 , is then crystallized or purified by flash column chromatography and carried onto the next step.
Preparation of 50
48 in nitromethane (.13 Molar), is added trimethylsilyl cyanide (3.0 equivalents) and then SnCl4 (.02 equivalents). The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the remaining trimethylsilyl cyanide. The mixture is evaporated and the remaining oil is resuspended in dichloromethane and washed with sodium acetate solution (1X), water (1X), brine (1X) and then dried over magnesium sulphate and concentrated. The crude solid is then recrystallized from methanol to yield 50 as a white solid (47%). scheme 4
Preparation of 52
The crude product 50 is next dissolved in ethanol (0.15 M) and then concentrated H2S O 4 (0.01 equivalents-catalytic) is added. The reaction mixture is heated to 85 °C for eight hours.
The solution is next concentrated in vacuo and purification by
flash column chromatography affords compound 52. scheme 4
Preparation of 54
A solution of 52 (1.0 equivalents) is dissolved in methanol (.1 M total). The reaction is then charged with acetic anhydride (1.1 equivalents) and is subsequently stirred for 2 hours at 30
°C. The reaction is next diluted with ether and washed with NaHC O3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 54. scheme 4
Preparation of 55
To a solution of 54 ( 1.0 equivalents) in pyridine (.10 Molar), is added trimethylacetylchloride (pivaloyl chloride) (2.5 equivalents) at 0 °C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine ( 1X), dried over MgSO4 and concentrated. Purification by flash column chromatography affords compound 55. scheme 4
Preparation of 56
To a solution of 55 ( 1.0 equivalents) in methylene chloride (.10 Molar), is added diisopropylethylamine (2.2 equivalents) at 0 ° C . S ub s e quent additi on of triethylsilyl trifluoromethanesulfonate (2.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 56. scheme 4
Preparation of 57
To a solution of 56 in ethanol (.13 Molar), is added sodium ethoxide (0.3 equivalents) and the reaction mixture is stirred for two hours at room temperature. The solution is then concentrated in vacuo and purification by flash column chromatography affords compound 57. scheme 4
Preparation of 58
A solution of 57 ( 1.0 equivalents) in tetrahydrofuran (.18 M) is treated with DPPA (diphenylphosphorylazide, 2.0 equivalents), triphenylphosphine ( 1.3 equivalents) and DIAD (diisopropyl-azo-dicarboxylate, 1.3 equivalents). The reaction is
heated to 80 °C for 3 hours and then diluted with ether (2X) and washed with .5 M aqueous NaOH (2X). The organic layer is dried over MgSO4 and evaporated. Purification by flash column chromatography affords compound 58. scheme 4
Preparation of 60
60
A solution of 58 ( 1.0 equivalents) is dissolved in ethanol (.01 M total) at 25 °C. The mixture is next exposed to 10% Pd/C (.1 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atm. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 60. scheme 4
Preparation of 61
A solution of 60 ( 1.0 equivalents) is dissolved in p- dioxanes (.1 M) and then exposed to a solution 3.0 Molar solution of sodium hydroxide (1.5 equivalents). The reaction is then stirred for 2 hours at 50 °C and is subsequently diluted with ether and washed with a solution of NH4CI (3X), brine (1X) and dried (MgSO4 ) and concentrated. Purification by flash column chromatography affords compound 61. scheme 4
Preparation of 62
To a solution of 61 ( 1.0 equivalents) in methylene chloride (.10 Molar), is added sodium bicarbonate (2.0 equivalents) at 0
°C. Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-Cl. 1.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried ( M g S O 4 ) and concentrated. Purification by flash column chromatography affords compound 62. scheme 4
Preparation of 63
To a stirred solution of the acid 46 ( 1.0 equivalents) and the amine 60 ( 1.1 equivalents) in dimethylformamide (.10 Molar) at 25 °C, is added 1-hydroxybenzotriazole (HOBT; 1.1 equivalents) . Next dicyclohexylcarbodiimide (1.2 equivalents) is added and the reaction is stirred for 14 hours. The mixture is diluted with ether, filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 63 .
scheme 5 step 1
Preparation of 64
To a stirred solution of 63 ( 1.0 equivalents) in dimethylformamide (.10 Molar) at 25 °C, is added piperidine (1.1 equivalents). The reaction is stirred for 1 hour and is then diluted with ether, and washed with aqueous CUSO4 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 64. scheme 5 step 2
Preparation of 65
To a stirred solution of the acid 62 (1.0 equivalents) and the amine 64 ( 1.1 equivalents) in dimethylformamide (.10 Molar) at 25 °C, is added 1-hydroxybenzotriazole (HOBT; 1.1 equivalents). Note: numerous iterations can be performed using the acid 62 or intermixing with other acids including for example acid 46 to form successive oligomers where n=2 to infinity (a hexamer is shown in scheme 5) to obtain large carbopeptoid libraries. Next dicyclohexylcarbodiimide ( 1.2 equivalents) is added and the reaction is stirred for 14 hours. The mixture is diluted with ether, filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 65. scheme 5 step 1
Preparation of 66
To a stirred solution of 65 ( 1.0 equivalents) in
dimethylformamide (.10 Molar) at 25 °C, is added piperidine (1.1 equivalents). The reaction is stirred for 1 hour and is then diluted with ether, and washed with aqueous CUSO4 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 66. No te : nume rous iterations can be performed using variable length oligomers of 66 to form peptoid oligomers where n=2 to infinity (a hexamer is shown in scheme 5). scheme 5 step 2
Preparation of 67
To a stirred solution of the acid 46 ( 1.0 equivalents) and the amine 66 ( 1.1 equivalents) in dimethylformamide ( .10 Molar) at 25 °C, is added 1-hydroxybenzotriazole (HOBT; 1.1 equivalents). Note: numerous iterations can be performed using the acid 46 or intermixing with other acids including for example acid 62, to form successive oligomers where n=2 to infinity (a hexamer is shown in scheme 5) to obtain large carbopeptoid libraries. Next dicyclohexylcarbodiimide ( 1.2
equivalents) is added and the reaction is stirred for 14 hours. The mixture is diluted with ether, filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 67. scheme 5 step 1
Preparation of 68
To a stirred solution of 67 ( 1.0 equivalents) in dimethylformamide (.10 Molar) at 25 °C, is added piperidine (1.1 equivalents). The reaction is stirred for 1 hour and is then diluted with ether, and washed with aqueous CUSO4 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 68. Note: nume rous iterations can be performed using variable length oligomers of 68 to form peptoid oligomers where n=2 to infinity (a hexamer is shown in scheme 5). scheme 5 step 2
Preparation of 69
To a stirred solution of the acid 62 ( 1.0 equivalents) and
the amine 68 ( 1.1 equivalents) in dimethylformamide (.10 Molar) at 25 °C, is added 1-hydroxybenzotriazole (HOBT; 1.1 equivalents). Note: numerous iterations can be performed using the acid 62, or intermixing with other acids including for example acid 46, to form successive oligomers where n=2 to infinity (a hexamer is shown in scheme 5) to obtain large carbopeptoid libraries. Next dicyclohexylcarbodiimide ( 1.2 equivalents) is added and the reaction is stirred for 14 hours. The mixture is diluted with ether, filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 69. scheme 5 step 1
Preparation of 70
To a stirred solution of 69 ( 1.0 equivalents) in dimethylformamide (.10 Molar) at 25 °C, is added piperidine ( 1.1 equivalents). The reaction is stirred for 1 hour and is then
diluted with ether, and washed with aqueous CUSO4 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 70. Note: n ume rous iterations can be performed using variable length oligomers of 70 to form peptoid oligomers where n=2 to infinity (a hexamer is shown in scheme 5). scheme 5 step 2
Preparation of 71
To a stirred solution of the acid 46 (1.0 equivalents) and the amine 70 ( 1.1 equivalents) in dimethylformamide (.10 Molar) at 25 °C, is added 1-hydroxybenzotriazole (HOBT; 1.1 equivalents). Note: numerous iterations can be performed using the acid 46 or intermixing with other acids including for example acid 62, to form successive oligomers where n=2 to infinity (a hexamer is shown in scheme 5) to obtain large carbopeptoid libraries. Next dicyclohexylcarbodiimide ( 1.2 equivalents) is added and the reaction is stirred for 14 hours. The mixture is diluted with ether, filtered and the filtrate is
washed with aqueous NaHCO3 (2X), water (2X), and brine (2X).
The organic phase is dried over MgSO4 and then concentrated.
Purification by flash column chromatography affords compound 71. scheme 5 step 1
Preparation of 72
To a stirred solution of 71 ( 1.0 equivalents) in dimethylformamide (.10 Molar) at 25 °C, is added piperidine (1.1 equivalents). The reaction is stirred for 1 hour and is then diluted with ether, and washed with aqueous CUSO4 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound 72. Note: numerous iterations can be performed using variable length oligomers of 72 to form peptoid oligomers where n=2 to infinity (a hexamer is shown in scheme 5). scheme 5 step 2
Preparation of 74
To a stirred solution of 72 ( 1.0 equivalents) in acetonitrile
(.50 Molar) is added an HF pyridine solution (.50 M) from Aldrich chemical company. The reaction is allowed to stir for five hours and is then condensed. The crude 73 oligomer is then resuspended in p-dioxane (.50 Molar) to which is added a 3.0 Molar solution of NaOH (3.0 equivalents). The reaction is stirred for 1 hour at 50 °C and is then quenched with aqueous NH4CI (2X) and subsequently lyophilized. Purification by HPLC chromatography affords compound 74. scheme 5
Preparation of 76
To a solution of β-D-Glucose pentaacetate 36 i n nitromethane from Aldrich company (.13 Molar), is added trimethylsilylcyanide (3.0 equivalents) and then tin tetrachloride (.02 equivalents). Note: other pyranose sugars such as β -D-Mannose, β-D-Galactose pentaacetate and other lewis acids such as BF3 O E t 2 may be used for alternative derivatives. The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the
remaining trimethylsilylcyanide. The mixture is evaporated and the remaining oil is resuspended in dichloromethane and washed with sodium acetate solution (1X), water (1X), brine (1X) and then dried over magnesium sulphate and concentrated. The crude product is next dissolved in ethanol (or methanol if the O- methyl glycoside is desired as in scheme 20), (0.15 M) and then concentrated H2S O 4 (0.01 equivalents) is added. The reaction mixture is heated to 85 °C for eight hours. The solution is next concentrated in vacuo and purification by flash column chromatography affords compound 76. scheme 6; 76, scheme 20 (as the O-methyl glycoside).
Preparation of 78
To Tetrol 76 (1.0 equivalents) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) (2.5 equivalents) at 0 °C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (1X), dried over MgSO4 and concentrated.
Next a solution of the crude intermediate ( 1.0 equivalents) is dissolved in methylene chloride ( . 10 Molar) and diisopropylethylamine (4.4 equivalents) is added at 0 °C. Subsequent addition of triethylsilyl trifluoromethanesulfonate (4.4 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSU4) and concentrated. Purification by flash column chromatography affords compound 78, scheme 6; 78, scheme 20 (as the O- methyl glycoside).
Preparation of 80
To a solution of 78 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added a 1.0 M solution of DIBALH in methylene chloride from Aldrich chemical company ( 1.2 equivalents) at 0 °C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with sodium-potassium tartrate (2X), brine (1X) and then MgSO4. The solution is then concentrated and purification by flash column chromatography affords
compound 80. scheme 6
Preparation of 82
82
To a solution of 80 (1.0 equivalents) in methylene chloride (.10 M), is added diisopropylethylamine (4.0 equivalents) at 25 °C. The reaction is stirred for 5 minutes and then 2-cyanoethyl- N, N-diisopropyl-chlorophosphoramidite ( 1.5 equivalents) is added, as prepared from the procedures of Sinha et al. Nu c l. Acids Res. 1984 , 12, 4539. After 15 minutes the reaction is complete and is next diluted with ether and next washed with brine ( 1X) and is then dried (MgSO4 ) and concentrated. Purification by flash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 82 (66% yield). scheme 6
Preparation of 84
To 80 ( 1.0 equivalents) in methylene chloride (.10 Molar) at 0 °C, is added diisopropylethylamine ( 1.1 equivalents). Subsequent addition of triethylsilyl trifluoromethanesulfonate (1.1 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. The crude is then resuspended in nitromethane and exposed to 10% CI3COOH ( 1.1 equivalents) in THF (.10 Molar). The reaction is stirred at 0 °C for 2 hours and is then diluted with ether and washed with sodium bicarbonate (2X), brine ( 1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 84. scheme 6
Preparation of 86
To a solution of N-phthalamido-D-Glucosamine tetraacetate
48 in nitromethane (.13 Molar), is added trimethylsilyl cyanide (3.0 equivalents) and then SnCl4 (.02 equivalents). The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the remaining trimethylsilyl cyanide. The mixture is evaporated and the remaining oil is resuspended in dichloromethane and washed with sodium acetate solution (1X), water (1X), brine (1X) and then dried over magnesium sulphate and concentrated. The crude product is next dissolved in ethanol (0.15 M) and then concentrated H2S O4 (0.04 equivalents) is added. The reaction mixture is heated to
85 °C for eight hours. The solution is next concentrated in vacuo and is then resuspended in methanol (.10 M) and acetic anhydride ( 1.1 equivalents) from Aldrich company is added in one step. After 2 hours, condensation and purification by flash column chromatography affords compound 86. scheme 7
Preparation of 88
To Triol 86 (1.0 equivalents) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) (2.5 equivalents) at 0 °C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (1X), dried over MgSO4 and concentrated. Next a solution of the crude intermediate ( 1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 °C. Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 88. scheme 7
Preparation of 90
To a solution of 88 ( 1.0 equivalents) in methylene chloride
(.10 Molar) is added a 1.0 M solution of DIBALH in methylene chloride from Aldrich chemical company (1.2 equivalents) at 0 °C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with sodium-potassium tartrate (2X), brine (1X) and then MgSO4. The solution is then concentrated and purification by flash column chromatography affords compound 90. scheme 7
Preparation of 92
To a solution of 90 ( 1.0 equivalents) in methylene chloride (.10 M), is added diisopropylethylamine (4.0 equivalents) at 25
°C. The reaction is stirred for 5 minutes and then 2-cyanoethyl- N, N-diisopropyl-chlorophosphoramidite ( 1.5 equivalents) is added, as prepared from the procedures of Sinha et al. Nu c l. Acids Res. 1984 , 12, 4539. After 15 minutes the reaction is complete and is next diluted with ether and next washed with brine (1X) and is then dried (MgSO4 ) and concentrated. Purification by flash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 92 (66% yield). scheme 7
Preparation of 94
To 90 ( 1.0 equivalents) in methylene chloride (.10 Molar) at 0 °C, is added diisopropylethylamine ( 1.1 equivalents).
Subsequent addition of triethylsilyl trifluoromethanesulfonate (1.1 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. The crude is then resuspended in nitromethane and exposed to 10% CI3COOH (1.1 equivalents) in THF (.10 Molar). The reaction is stirred at 0 °C for 2 hours and is then
diluted with ether and washed with sodium bicarbonate (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 9 4 . scheme 7
Preparation of 98 (homodimer scheme 8)
To a solution of 94 (1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company ( 10.0 equivalents) at 25 °C. Next, a solution of 82 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at
25 °C. After 25 minutes, the mixture is cooled to 0 °C and I2 (4.0 equivalents), 2,6 lutidine (4.0 equivalents) in TΗF ( 1.0 M) is added to oxidize the phosphoamidate to the phosphate (Alternatively m-chloroperoxybenzoic acid (4.5 equivalents) is added). The reaction is next stirred for an additional 5 minutes and is next diluted with ether and washed with brine (1X) and dried (MgSO4 ) and concentrated. Purification by flash column chromatography and then the product is suspended in acetic acid-tetrahydrofuran-water (3: 1 : 1 ), (.01 M) and stirred for 18 hours at 25 °C. The reaction is then diluted with ether and washed with NaΗCO3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 98 (scheme 8).
Preparation of 102 (heterotrimer scheme 8)
To a solution of 98 (1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company ( 10.0 equivalents) at 25 °C. Next, a solution of 92 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at
25 °C. After 25 minutes, the mixture is cooled to 0 °C and I2 (4.0 equivalents), 2,6 lutidine (4.0 equivalents) in TΗF (1.0 M) is added to oxidize the phosphoamidate to the phosphate (Alternatively m-chloroperoxybenzoic acid (4.5 equivalents) is added). The reaction is next stirred for an additional 5 minutes and is next diluted with ether and washed with brine (1X) and dried (MgSO4 ) and concentrated. Purification by flash column chromatography and then the product is suspended in acetic acid-tetrahydrofuran-water (3: 1 : 1), (.01 M) and stirred for 18 hours at 25 °C. The reaction is then diluted with ether and washed with NaΗCO3 (3X), brine (1X) and dried (MgSO4 ) and concentrated. Purification by flash column chromatography affords compound 102 (scheme 8). Preparation of 106 (hetero tetramer scheme 8)
To a solution of 102 ( 1.0 equivalents) in methylene chloride (.10 M), is added 1 -H-tetrazole from Aldrich company ( 10.0 equivalents) at 25 °C. Next, a solution of 82 (3.0 equivalents) in methylene chloride (1.0 M), is added dropwise
with stirring at 25 °C. After 25 minutes, the mixture is cooled to 0 °C and I2 (4.0 equivalents), 2,6 lutidine (4.0 equivalents) in THF ( 1.0 M) is added to oxidize the phosphoamidate to the phosphate (Alternatively m-chloroperoxybenzoic acid (4.5 equivalents) is added). The reaction is next stirred for an additional 5 minutes and is next diluted with ether and washed with brine ( I X) and dried (MgSO4 ) and concentrated. Purification by flash column chromatography and then the product is suspended in acetic acid-tetrahydrofuran-water (3: 1 : 1), (.01 M) and stirred for 18 hours at 25 °C. The reaction is then diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 106 (scheme 8). Preparation of 110 (heteropentamer scheme 8)
To a solution of 106 ( 1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company ( 10.0 equivalents) at 25 °C. Next, a solution of 92 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at 25 °C. After 25 minutes, the mixture is cooled to
0 °C and I2 (4.0 equivalents), 2,6 lutidine (4.0 equivalents) in TΗF ( 1.0 M) is added to oxidize the phosphoamidate to the phosphate (Alternatively m-chloroperoxybenzoic acid (4.5 equivalents) is added). The reaction is next stirred for an
additional 5 minutes and is next diluted with ether and washed with brine ( I X) and dried (MgSO4 ) and concentrated. Purification by flash column chromatography and then the product is suspended in acetic acid-tetrahydrofuran-water (3: 1 : 1), (.01 M) and stirred for 18 hours at 25 °C. The reaction is then diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 110 (scheme 8). Preparation of 114 (heterohexamer scheme 8)
To a solution of 110 ( 1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company (10.0 equivalents) at 25 °C. Next, a solution of 82 (3.0 equivalents) in methylene chloride (1.0 M), is added dropwise with stirring at 25 °C. After 25 minutes, the mixture is cooled to
0 °C and I2 (4.0 equivalents), 2,6 lutidine (4.0 equivalents) in TΗF ( 1.0 M) is added to oxidize the phosphoamidate to the phosphate (Alternatively m-chloroperoxybenzoic acid (4.5 equivalents) is added). The reaction is next stirred for an additional 5 minutes and is next diluted with ether and washed with brine ( I X) and dried (MgSO4 ) and concentrated. Purification by flash column chromatography and then the product is suspended in acetic acid-tetrahydrofuran-water (3: 1 : 1 ), (.01 M) and stirred for 18 hours at 25 °CC The reaction is
then diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 114 (scheme 8). Preparation of 116 (heterohexamer scheme 8)
To a solution of 114 (1.0 equivalents) in methylene
chloride (.10 M), is added a solution of HF-pyridine (1.0 M) at 0
°C. The reaction is next stirred for an additional 30 minutes and
is next diluted with ether and washed with a saturated solution
of sodium bicarbonate (3X), copper sulfate solution to remove
the pyridine (2X) brine (1X), dried (MgSO4) and concentrated.
Purification by flash column chromatography and then the
product is resuspended in concentrated aqueous ammonium
hydroxide and acetonitrile (1: 1), (.1 M total). The reaction is
then stirred for 2 hours at 50 °C and is subsequently diluted
with ether and washed with NaHCO3 (3X), brine (1X) and dried
(MgSO4) and concentrated. Purification by flash column
chromatography affords compound 116 scheme 8.
To a solution of β-D-Glucose pentaacetate in nitromethane from Aldrich company ( . 1 3 M olar) , i s added trimethylsilylcyanide (3.0 equivalents) and then borontrifluoride etherate (.02 equivalents). Note: other pyranose sugars such as β -D-Mannose, β -D-Galactose pentaacetate and other lewis acids such as SnCl4, may be used for alternative derivatives. The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the remaining trimethylsilylcyanide. The mixture is evaporated and the remaining oil is resuspended in dichloromethane and washed with sodium acetate solution (1X), water ( 1X), brine (1X) and then dried over magnesium sulphate and concentrated. The crude solid is then recrystallized from methanol to yield 125 (also 37) as a white solid (47%). scheme 9 step a
Preparation of 126
To a solution of 125 in methanol (.13 Molar), is added sodium methoxide (0.3 equivalents) and the reaction mixture is stirred for two hours at room temperature. The dark brown solution is then concentrated in vacuo to give a dark brown syrup of compound 126 which is carried on without purification as a crude oil for the next step, scheme 9 step b
Preparation of 127
The crude product 126 is dissolved in 25% NaOH (0.5 M) and heated at reflux for 18 hours (vigorous reflux is necessary). Next, the solution is diluted with an addition of water (0.1 M) and to this solution is added Amberlite 112120 resin (H+-form) and is then stirred. The supernatant is then decanted and the
resin is washed until the eluate is colorless. The eluate is then collected, condensed and azeotroped with MeOH which yields 127 as a crude, pale yellow syrup (47%).
Preparation of 130
The crude product 127 is next dissolved in methanol (0.15 M) and then concentrated HCl (0.01 equivalents) is added. The reaction mixture is heated to 85 °C for eight hours. The solution is next concentrated in vacuo and purification by flash column chromatography (silica, 20% methanol in ethyl acetate), affords compound 130 as a white solid (60% yield), scheme 9 step d
Preparation of 131
To a solution of 130 ( 1 .0 equivalents ) in dimethylformamide (.23 Molar), is added imidazole (2.5 equivalents) at 0 °C. Subsequent addition of te rt- B u ty l - dimethylsilylchloride (2.5 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4 ) and concentrated. Purification by flash column chromatography (silica, 50% ethyl acetate) affords compound 131 as a white solid (93% yield), scheme 9 step e Note: the molecule can be protected with other primary directing protecting groups such as DMT (dimethoxytrityl), and TBDPS tert-butyldiphenlysilyl, etc. Preparation of 132
To a solution of 1 3 1 ( 1 .0 equivalents ) in dimethylformamide (.23 M), is added Ag2O (6.0 equivalents) at 25 °C. Benzyl bromide (9.0 equivalents) is next added and the
reaction is allowed to stir for 20 hours. The reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4 ) and concentrated. Purification by flash column chromatography (silica, 20% ethyl acetate) affords compound 132 (83% yield), scheme 9 step f Note: the choice of the protecting group is relative and the molecule can be protected with other protecting groups at C2, C3, C4, such as PMB (paramethoxybenzyl), TES (triethylsilyl), TBS (tertbutyldimethylsilyl), etc.
Preparation of 134
To a solution of 132 ( 1.0 equivalents) in tetrahydrofuran
(.08 M), is added diisobutylaluminumhydride (DIBALH) (3.0 equivalents) at 0 °C. The reaction is stirred for 1 hour and then quenched with methanol and diluted with ether. The reaction is next worked-up with ammonium chloride (2X), brine (1X) and is then dried (MgSO4 ) and concentrated. Purification by flash column chromatography (silica, 20% ethyl acetate) affords compound 134 (66% yield), scheme 9 step g
Preparation of 136
To a solution of 134 ( 1.0 equivalents) in pyridine ( 10.0 equivalents), is added naphthoyl chloride (3.0 equivalents) from Aldrich company (3.0 equivalents) at 25 °C. The reaction is stirred for 45 minutes and then diluted with ether and worked- up with a saturated solution of CUSO4 (2X), brine (1X) and is then dried (MgSO4) and concentrated. The crude product is then exposed to acetic acid/tetrahydrofuran/water (3 : 1 : 1 ) at 25 °C and allowed to stir for 15 hours. The reaction is then diluted with ether and worked-up with brine (2X) and is then dried ( M g S O 4 ) and concentrated. Purification by flash column chromatography (silica, 20% ethyl acetate) affords compound 136 (95% yield). Note: alternatively, one could originally protect the C7 position as a DMT (dimethoxytrityl) functionality and protect the C1 position as a TES (triethyl silyl) group. Subsequent mild acid hydrolysis of the DMT group leads to the
above compound with the TES group at the C1 position and a free hydroxyl at the C7 position, scheme 9 step h
Preparation of 138
138
To a solution of 134 ( 1.0 equivalents) in methylene chloride ( .10 M), is added diisopropylethylamine (4.0 equivalents) at 25 °C. The reaction is stirred for 5 minutes and then 2-cyanoethyl-N, N-diisopropyl-chlorophosphoramidite ( 1.5 equivalents) is added, as prepared from the procedures of Sinha et al. Nucl. Acids Res. 1984 , 12, 4539. After 15 minutes the reaction is complete and is next diluted with ether and next washed with brine (1X) and is then dried (MgSO4 ) and concentrated. Purification by flash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 138 (66% yield), scheme 9 step i
It should be noted that the oligomerization process as shown below in scheme 9, uses the same C-glycoside 138 in an iterative fashion. The process can be
extended however to include a pool of random or ordered C-glycosides as depicted in scheme 8.
Preparation of 140
To a solution of 136 ( 1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company ( 10.0 equivalents) at 25 °C. Next, a solution of 138 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at 25 °C. After 25 minutes, the mixture is cooled to 0 °C and m-chloroperoxybenzoic acid (4.5 equivalents) is added. The reaction is stirred for an additional 5 minutes and is next diluted with ether and washed with brine (1X) and dried ( M g S O 4 ) and concentrated. Purification by flash column chromatography (silica, 50% ethyl acetate in petroleum ether) affords compound 140 (97% yield), scheme 9 step j Note the process can iterate as many times as possible to build large carbonucleotide libraries.
Preparation of 142
A solution of 140 ( 1.0 equivalents) in acetic acid- tetrahydrofuran-water (3: 1 : 1 ), (.01 M) is stirred for 18 hours at 25 °C. The reaction is then diluted with ether and washed with NaHC O 3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography (silica, 60% ethyl acetate in petroleum ether) affords compound 142 (95% yield). scheme 9 step k Note the process can iterate as many times as possible to build large carbonucleotide libraries.
Preparation of 144
To a solution of 138 ( 1.0 equivalents) in methylene chloride (.10 M), is added 1-H-tetrazole from Aldrich company
( 10.0 equivalents) at 25 °C. Next, a solution of 142 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at 25 °C. After 25 minutes, the mixture is cooled to 0 °C and m-chloroperoxybenzoic acid (4.5 equivalents) is added. The reaction is stirred for an additional 5 minutes and is next diluted with ether and washed with brine (1X) and dried ( M g S O 4 ) and concentrated. Purification by flash column chromatography (silica, 50% ethyl acetate in petroleum ether) affords compound 144 (97% yield), scheme 9 step j Note the process can iterate as many times as possible to build large carbonucleotide libraries.
Preparation of 146
A solution of 144 ( 1.0 equivalents) in acetic acid- tetrahydrofuran-water (3: 1 : 1 ), (.01 M total) is stirred for 18 hours at 25 °C. The reaction is then diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography (silica, 60% ethyl acetate in petroleum ether) affords compound 146 (95% yield), scheme 9 step k Note the process can iterate as many times as possible to build large carbonucleotide libraries.
Preparation of 148
To a solution of 138 ( 1.0 equivalents) in methylene
chloride (.10 M), is added 1-H-tetrazole from Aldrich company ( 10.0 equivalents) at 25 °C. Next, a solution of 146 (3.0 equivalents) in methylene chloride ( 1.0 M), is added dropwise with stirring at 25 °C. After 25 minutes, the mixture is cooled to 0 °C and m-chloroperoxybenzoic acid (4.5 equivalents) is added.
The reaction is stirred for an additional 5 minutes and is next diluted with ether and washed with brine (1X) and dried ( M g S θ 4 ) and concentrated. Purification by flash column chromatography (silica, 50% ethyl acetate in petroleum ether) affords compound 148 (97% yield), scheme 9 step j Note the process can iterate as many times as possible to build large carbonucleotide libraries.
A solution of 1 48 ( 1.0 equivalents) in acetic acid- tetrahydrofuran-water (3: 1 : 1 ), (.01 M total) is stirred for 18 hours at 25 °C. The reaction is then diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4 ) and concentrated. Purification by flash column chromatography (silica, 60% ethyl acetate in petroleum ether) affords compound 150 (95% yield). scheme 9 step k Note the process can iterate as many times as possible to build large carbonucleotide
libraries.
A solution of 1 50 ( 1.0 equivalents) is dissolved in concentrated aqueous ammonium hydroxide and acetonitrile (1 : 1), (.1 M total). The reaction is then stirred for 2 hours at 50 °C and is subsequently diluted with ether and washed with NaHCO 3 (3X), brine (1X) and dried (MgSO4) and concentrated.
Purification by flash column chromatography (silica, 80% ethyl acetate in petroleum ether) affords compound 152 (88% yield). scheme 9 step L Preparation of 154
A solution of 1 52 ( 1.0 equivalents) is dissolved in a mixture of ethanol-tetrahydrofuran-acetic acid (2: 1 : 1 ), (.01 M total) at 25 °C. The mixture is next exposed to 10% Pd/C ( 1.0 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atmosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is
subsequently diluted with ether and washed with NaHCθ3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography (silica, 100% ethyl acetate in petroleum ether) affords compound 154 (78% yield), scheme 9 step m
Preparation of 174 (R group = OTES, NPhth or NHAc)
To a solution of tetraacetate derived from 36 or 48 (glucose or glucosamine derived) in methylene chloride (.1 molar) is added a 1.0 molar solution of Co2(CO)8 (1.5 equivalents ) in methylene chloride and diethylmethylsilane (1.5 equivalents) at 0 °C. To the stirring reaction mixture, a stream of carbon monoxide is bubbled at 1 ml per 10 seconds for 30 minutes. The reaction mixture is then quenched with water (1.5 equivalents), diluted with ether, washed with sodium bicarbonate (2x), brine ( lx) and dried over magnesium sulfate. The crude is purified by column chromatography and affords product 174.
Preparation of 176 (R group = OTES, NPhth or NHAc)
To a solution of compound 174 in acetonitrile/water (1 : 1 ratio,
.1 molar combined), is added RuCl3 (.03 equiv.) and NaIO4 (4.0 equiv.) at 25 °C and the muddy black mixture is allowed to stir for 1.5 h. The mixture is then diluted with ether (25 mL), washed with water (2X 5.0 mL) and brine ( IX 5 mL). The
aqueous layer is back extracted (2X), recombined, and the organic layer was then dried MgSO4 and evaporated. Purification by flash column chromatography yields the desired product 176.
Preparation of 178 (R group = OTES, NPhth or NHAc)
A solution of triacetate 176 ( 1.0 equiv.) in methanol (0.5 M), is treated with NaOMe (0.4 equiv.) and allowed to stir at 25 °C for 24 h. The reaction mixture is then condensed and purified by flash column chromatography to afford compound 178.
Preparation of 180 (R group = OTES, NPhth or NHAc)
To triol 178 (1.0 equivalents) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) (1.5 equivalents) at 0 °C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (1X), dried over MgSO4 and concentrated. Next a solution of the crude intermediate ( 1.0 equivalents) is dis s olved in methylene chloride ( .10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 °C.
Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and
concentrated. Purification by flash column chromatography affords the intermediate acid, which is then resuspended in THF ( 1.0 M) and exposed to a 1.0 M solution of BH3 -THF ( 1.5 equivalents) at 0 °C for 1 hour. The reaction is then quenched with methanol for an additional hour and the crude is then diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4 ) and concentrated. Purification by flash column chromatography affords the desired tetraprotected alcohol 180.
Preparation of 181 (R group = OTES, NPhth or NHAc)
To a solution of 180 (1.0 equivalents) in methylene chloride (.10 M), is added tetrazole (4.0 equivalents) at 25 °C. The reaction is stirred for 5 minutes and then 2-cyanoethyl-N, N-dii sopropyl- chlorophosphoramidite ( 1.5 equiv.) is added, as prepared from the procedures of Sinha et al. Nucl. Acids Res. 1984 , 12, 4539. After 15 minutes the reaction is complete and is next diluted with ether and next washed with brine (1X) and is then dried ( M g S O 4 ) and concentrated. Purification by flash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 181 (66% yield), scheme 21
Preparation of 182 (R group = OTES, ΝPhth or ΝHAc)
To a solution of triol 178 (.0 equiv.) in CH2CI2 (.5 M) at 0 °C, was
added triethylamine ( 1.2 equiv.), 4-DMAP (.10 equiv.) and then TOSCl (1.1 equiv.). The reaction is stirred for 1 h and then is quenched with saturated ammonium chloride ( 1.5 mL), diluted with ethyl acetate (25 mL), washed with water (2X 5 mL), brine (1X 5 mL), back-extracted (2X), recombined, dried (MgSO4 ) and evaporated. The compound is purified by flash column chromatography and then a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (2.2 equivalents) is added at 0 °C. Subsequent addition of triethylsilyl trifluoromethanesulfonate
(2.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords the protected tosylate/acid 182.
Preparation of 183 (R group = OTES, NPhth or NHAc)
To a solution of triol 182 (.0 equiv.) in CH2CI2 (.5 M) at 0 °C, is added sodium-azide (1.2 equiv.) from Aldrich chemical company at 0 °C. The reaction is stirred for 1 h and then is quenched with saturated ammonium chloride ( 1.5 mL), diluted with ethyl acetate (25 mL), washed with water (2X 5 mL), brine (1X 5 mL), back-extracted (2X), recombined, dried (MgSO4 ) and evaporated. The compound is purified by flash column chromatography and
affords compound 183.
Preparation of 185 (R group = OTES, NPhth or NHAc)
A solution of 183 (1.0 equivalents) in ethanol (.01 M total) at 25 °C is exposed to 10% Pd(OH)2 -C (0.1 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atmosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 185 scheme 21.
Preparation of 191
A solution of starting material 1 90 as disclosed by Schmidt, R. R. et al. (Liebigs Ann. Chem. 1 987 , 825), ( 1.0 equivalents) is dis solved in a mixture of ethanol- tetrahydrofuran-acetic acid (2: 1 : 1), (.01 M total) at 25 °C. The mixture is next exposed to 10% Pd/C ( 1.0 equivalents) and is
then subsequently capped with a hydrogen balloon at 1 atmosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography (silica, 100% ethyl acetate in petroleum ether) affords compound 191. scheme 22 step a
Preparation of 192
To a solution of 191 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added tosylchloride (1.2 equivalents) at 0 °C. Subsequent addition of triethylamine ( 1.5 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated to afford the crude tosylate. Next a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 °C. Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 192. scheme 22 step b
Preparation of 193
To a solution of 192 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added sodium azide from Aldrich chemical company (1.2 equivalents) at 0 °C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with ammonium chloride (2X), brine (1X) and then M g S O4. The solution is then concentrated and purification by flash column chromatography affords compound 193. scheme
22 step c
Preparation of 194
To solution of 193 in CCI4 (.33 M), CH3CN (.33 M) and water (.22 M) at 0 °C is added RuCl3 (.03 equiv.) and NaIO4 (4.0 equiv.) and the muddy black mixture is allowed to stir for 1.5 h. The mixture is then diluted with ether (25 mL), washed with
water (2X 5.0 mL) and brine (IX 5 mL). The aqueous layer is back extracted (2X), recombined, and the organic layer iss then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 194. scheme 22 step d
Preparation of 196
A solution of 194 (1.0 equivalents) is dissolved in ethanol (.01 M total) at 25 °C. The mixture is next exposed to 10% Pd/C (.1 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atmosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4 ) and concentrated. Next, to a solution of crude amine ( 1.0 equivalents) in methylene chloride (.10 Molar), is added sodium bicarbonate (2.0 equivalents) at 0
°C. Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-Cl, 1.2 equivalents) is followed by stirring for 2 hours
and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 196. scheme 22 steps e-f
Preparation of 197
To Tetrol 191 (1.0 equivalents) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) (2.5 equivalents) at 0 °C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (1X), dried over MgSO4 and concentrated. Next a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride ( .10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 °C. Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 197. scheme 22 step g
Preparation of 198
To solution of 197 in CCI4 (.33 M), CH3CN (.33 M) and water (.22 M) at 0 °C is added RuCl3 (.03 equiv.) and NaIO4 (4.0 equiv.) and the muddy black mixture is allowed to stir for 1.5 h. The mixture is then diluted with ether (25 mL), washed with water (2X 5.0 mL) and brine ( IX 5 mL). The crude is then resuspended in a mixture of methylene chloride/water (1 : 1 , .1 M total) and diazomethane ( 1.2 equivalents) is gradually dropped into the flask via an addition funnel at the rate of 1 drop/10 seconds. After complete addition the mixture is diluted with ether, washed with brine (2X) and the aqueous layer is back extracted (2X) recombined, and the organic layer is then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 198. scheme 22 step h
Preparation of 200
To a solution of 198 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added a 1.0 M solution of DIBALH in methylene chloride from Aldrich chemical company ( 1.2 equivalents) at 0 °C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with sodium- potassium tartrate (2X), brine (1X) and then MgSO4. The solution is then concentrated and purification by flash column chromatography affords compound 200. scheme 22 step i
Preparation of 201
A solution of 200 ( 1.0 equivalents) in tetrahydrofuran (.18 M) is treated with DPPA (diphenylphosphorylazide, 2.0
equivalents), triphenylphosphine ( 1.3 equivalents) and DIAD (diisopropyl-azo-dicarboxylate, 1.3 equivalents). The reaction is heated to 80 °C for 3 hours and then diluted with ether (2X) and washed with .5 M aqueous NaOH (2X). The organic layer is dried over MgSO4 and evaporated. Purification by flash column chromatography affords compound 201. scheme 22 step j
Preparation of 202
A solution of 201 (1.0 equivalents) is dissolved in ethanol (.01 M total) at 25 °C. The mixture is next exposed to 10% Pd/C (.1 equivalents) and is then subsequently capped with a hydrogen balloon at 1 atmosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4 ) and concentrated. Next, to a solution of crude amine ( 1.0 equivalents) in methylene chloride (.10 Molar), is added sodium bicarbonate (2.0 equivalents) at 0
°C. Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-Cl, 1.2 equivalents) is followed by stirring for 2 hours
and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried ( M g S O 4 ) and concentrated. Purification by flash column chromatography affords compound 202. scheme 22 step e
Preparation of 204
To a solution of 202 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added 10% HCOOH from Aldrich chemical company ( 1.2 equivalents) at 0 °C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with sodium bicarbonate (2X), brine (1X) and then MgSO4. The solution is then resuspended in t-BuOH (.10 M) and pH 7 buffer
(.10 M) and is then exposed to KMnO4 ( 1.2 equivalents) for 2 hours at 0 °C. The reaction mixture is next washed with sodium bicarbonate (2X), brine (1X) and then MgSO4. The organic layer is then concentrated and purified by flash column chromatography to afford compound 2044 scheme 22 step k
Preparation of 206
To Tetrol 205 (1.0 equivalents) (as disclosed by Petrus, L. et al.Chem. zvesti. 1982 , 36 , 103) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) (2.5 equivalents) at 0 °C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (1X), dried over MgSO4 and concentrated.
Next a solution of the crude intermediate ( 1.0 equivalents) is dissolved in methylene chloride ( .10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 °C. Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 206. scheme 23 step a
Preparation of 207
To a solution of 206 ( 1.0 equivalents) in diethylether (.08
M), is added lithiumaluminumhydride (LAH) ( 1.5 equivalents) at 30 °C. The reaction is refluxed for 2 hours and then quenched with methanol and diluted with ether. The reaction is next worked-up with sodium potassium tartrate (2X), brine (1X) and is then dried (MgSO4) and concentrated. The crude mixture is resuspended in methylene chloride (.10 Molar) and to it is added sodium bicarbonate (2.0 equivalents) at 0 °C.
Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC- Cl, 1.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 207. scheme 23 step b
Preparation of 208
To a solution of 207 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added 10% HCOOH (1.1 equivalents). The reaction is stirred at 0 °C for 2 minutes and is then diluted with ether and washed with sodium bicarbonate (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 208. scheme 23 step c
Preparation of 209
To solution of 208 in CCI4 (.33 M), CH3 CN (.33 M) and water (.22 M) at 20 °C is added RuCl3 (.03 equiv.) and NaI04
(4.0 equiv.) and the muddy black mixture is allowed to stir for
10 min. The mixture is then diluted with ether (25 mL), washed with water (2X 5.0 mL) and brine (IX 5 mL). The aqueous layer
is back extracted (2X), recombined, and the organic layer iss then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 209. scheme 23 step d
Preparation of 210
To a solution of 205 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added tosylchloride ( 1.2 equivalents) at 0 °C. Subsequent addition of triethylamine ( 1.5 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated to afford the crude tosylate. Next a solution of the crude intermediate (1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 °C. Subsequent addition of triethylsilyl trifluoromethanesulfonate
(3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with
ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 210. scheme 23 step e
Preparation of 211
To a solution of 210 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added sodium azide from Aldrich chemical company (1.2 equivalents) at 0 °C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with ammonium chloride (2X), brine (1X) and then M g S O4. The solution is then concentrated and purification by flash column chromatography affords compound 211. scheme
23 step f
Preparation of 212
To solution of 211 in CCI4 (.33 M), CH3CN (.33 M) and water (.22 M) at 20 °C is added RuCl3 (.03 equiv.) and NaI04 (4.0 equiv.) and the muddy black mixture is allowed to stir for 10 min. The mixture is then diluted with ether (25 mL), washed with water (2X 5.0 mL) and brine (IX 5 mL). The aqueous layer is back extracted (2X), recombined, and the organic layer iss then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 212. scheme 23 step
Preparation of 213
A solution of 212 ( 1.0 equivalents) in ethanol (.01 M total) at 25 °C is exposed to 10% Pd/C (0.1 equivalents) and is then
subsequently capped with a hydrogen balloon at 1 atmosphere. The reaction is stirred for 72 hours and is then filtered through celite. The crude mixture is subsequently diluted with ether and washed with NaHCO3 (3X), brine (1X) and dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 213. scheme 23 step h
Preparation of 214
Compound 213 is suspended in methylene chloride (.10 Molar) and to it is added sodium bicarbonate (2.0 equivalents) at 0 °C . Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-Cl, 1.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 214. scheme 23 step i
Preparation of 215
To a solution of 205 ( 1.0 equivalents) in pyridine (.10
Molar), is added trimethylacetyl chloride (pivaloyl chloride) (2.5 equivalents) at 0 °C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (1X), dried over MgSO4 and concentrated. Next a solution of the crude intermediate
(1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 °C. Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 215. scheme 23 step j
Preparation of 216
To solution of 215 in CCI4 (.33 M), CH3 CN (.33 M) and water (.22 M) at 20 °C is added RuCl3 (.03 equiv.) and NaIO4 (4.0 equiv.) and the muddy black mixture is allowed to stir for 10 min. The mixture is then diluted with ether (25 mL), washed with water (2X 5.0 mL) and brine (IX 5 mL). The aqueous layer is back extracted (2X), recombined, and the organic layer is then dried MgSO4 and evaporated. The crude is then resuspended in a mixture of methylene chloride/water ( 1 : 1 , .1 M total) and diazomethane ( 1.2 equivalents) is gradually dropped into the flask via an addition funnel at the rate of 1 drop/10 seconds. After complete addition the mixture is diluted with ether, washed with brine (2X) and the aqueous layer is back extracted (2X) recombined, and the organic layer is then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 216. scheme 23 step k
Preparation of 217
To a solution of 216 ( 1.0 equivalents) in methylene chloride (.10 Molar) is added a 1.0 M solution of DIBALH in methylene chloride from Aldrich chemical company ( 1.2 equivalents) at 0 °C. Subsequent stirring for 2 hours is followed by dilution with diethylether and washing with sodium- potassium tartrate (2X), brine (1X) and then MgSO4. The solution is then concentrated and purification by flash column chromatography affords compound 217. scheme 23 step 1
Preparation of 218
To 217 (1.0 equivalents) in pyridine (.10 Molar), is added dimethyoxytritylchloride (DMT chloride) (1.1 equivalents) at 0°C. The reaction is stirred for 2 hours and then diluted with
diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (1X), dried over MgSO4 and concentrated. Purification by flash column chromatography affords compound 218. scheme 23 step m
Preparation of 220
To a solution of 218 ( 1.0 equivalents) in diethylether (.08
M), is added lithiumaluminumhydride (LAH) (1.5 equivalents) at 30 °C. The reaction is refluxed for 2 hours and then quenched with methanol and diluted with ether. The reaction is next worked-up with sodium potassium tartrate (2X), brine (1X) and is then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 220. scheme 23 step n
Preparation of 221
To Tetrol 205 ( 1.0 equivalents) in pyridine (.10
Molar), is added dimethyoxytritylchloride (DMT chloride) (2.5 equivalents) at 0 °C. The reaction is stirred for 2 hours and then diluted with diethylether and washed with ammonium chloride (2X), copper sulfate (2X), brine (1X), dried over MgSO4 and concentrated. Next a solution of the crude intermediate ( 1.0 equivalents) is dissolved in methylene chloride (.10 Molar) and diisopropylethylamine (3.3 equivalents) is added at 0 °C. Subsequent addition of triethylsilyl trifluoromethanesulfonate (3.3 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 221. scheme 23 step a
Preparation of 222
To solution of 221 in CCI4 (.33 M), CH3CN (.33 M) and water (.22 M) at 20 °C is added RuCI3 (.03 equiv.) and NaIO4 (4.0 equiv.) and the muddy black mixture is allowed to stir for 10 min. The mixture is then diluted with ether (25 mL), washed with water (2X 5.0 mL) and brine (1X 5 mL). The aqueous layer is back extracted (2X), recombined, and the organic layer iss then dried MgSO4 and evaporated. Purification by flash column chromatography affords the compound 222. scheme 23 step o.
Preparation of 224
To a solution of 222 ( 1.0 equivalents) in diethylether (.08
M), is added lithiumaluminumhydride (LAH) ( 1.5 equivalents) at 30 °C. The reaction is refluxed for 2 hours and then quenched with methanol and diluted with ether. The reaction is next worked-up with sodium potassium tartrate (2X), brine (1X) and is then dried (MgSO4) and concentrated. Purification by flash column chromatography affords compound 224. scheme 23 step p
Preparation of 216
To a stirred solution of the acid 214 ( 1.0 equivalents) in dimethylformamide (.10 Molar) at 25 °C, is added 1 - hydroxybenzotriazole (HOB T ; 1 . 1 equivalents ) . Next dicyclohexylcarbodiimide ( 1.2 equivalents) is added and the reaction is stirred for 1 hour in the presence of an appropriately substituted solid support (N-(2-Aminoethyl)-3-amino-propyl glass; aminopolystyrene resin; aminopropyl glass; isothiocyanato glass, all with or without a linker extending from the amino group on the support etc. from Sigma Company). The mixture is then diluted with ether, filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated.
Preparation of 226; 228; 230 or 232
To a stirred solution of the acid 214; 62; 215 or 62 ( 1 . 0
equivalents) and the amine 216; 226; 228 or 230 ( 1.1 equivalents) in dimethylformamide (.10 Molar) at 25 °C, is added 1-hydroxybenzotriazole (HOBT; 1.1 equivalents). N e x t dicyclohexylcarbodiimide (1.2 equivalents) is added and the reaction is stirred for 14 hours. The mixture is diluted with ether, filtered and the filtrate is washed with aqueous NaHCO3 (2X), water (2X), and brine (2X). The organic phase is dried overMgSO4 and then concentrated. Purification by flash column chromatography and then reexposure of the intermediate compound ( 1.0 equivalents) in dimethyl-formamide (.10 Molar) at 25 °C, is added piperidine (1.1 equivalents). The reaction is stirred for 1 hour and is then diluted with ether, and washed with aqueous CUSO4 (2X), water (2X), and brine (2X). The organic phase is dried over MgSO4 and then concentrated. Purification by flash column chromatography affords compound
226; 228; 230 or 232, respectively, scheme 24
Preparation of 234
To a stirred solution of 232 ( 1.0 equivalents) in acetonitrile (.50 Molar) is added an HF-pyridine solution (.50 M) from Aldrich chemical company. The reaction is allowed to stir for five hours and is then condensed. The crude 234 oligomer is then resuspended in p-dioxane (.50 Molar) to which is added a 3.0 Molar solution of NaOH (3.0 equivalents). The reaction is stirred
for 1 hour at 50 °C and is then quenched with aqueous NH4C I (2X) and subsequently lyophilized. Purification by HPLC chromatography affords compound 234. scheme 24
Preparation of Peptoid Combinatorial libraries
(Scheme 500)
A depiction of the generation of a combinatorial library for oligopeptoid compounds is shown in scheme 500. The example uses an alphabet of eight D-aldose hexose sugars (other sugars groups such as the D/L ketoses and L-configurations of aldose hexoses, may be used) and carries the synthesis to a degree of three or 512 compounds. (The process can repeat itself to afford the library of desired size). Standard chemistry is shown and follows the reaction conditions as described above herein for peptoid synthesis. The solid support used is the standard N-(2- Aminoethyl)-3-amino-propyl glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support
(eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
Preparation of Nucleotoid Combinatorial libraries (Scheme 550)
A depiction of the generation of a combinatorial library for oligonucleotoid compounds is shown in scheme 550. The example uses an alphabet of eight D-aldose hexose sugars (other sugars groups such as the D/L ketoses and L-configurations of aldose hexoses, may be used) and carries the synthesis to a degree of three or 512 compounds. (The process can repeat itself to afford the library of desired size). Standard chemistry is shown and follows the reaction conditions as described above herein for carbonucleotoid synthesis. The solid support used is the standard N-(2-Aminoethyl)-3-amino-propyl glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
Preparation of compound 2000. To a solution of 76 ( 1.0 equiv) was added methylene chloride (.1 M) and benzaldehyde ( 1.1 equiv), and the solution was exposed to ZnCl ( 1.1 equiv) at 25 °C and allowed to stir for 2.5 hour. The solution is then
diluted with ether and then washed with a saturated solution of sodium bicarbonate (2X), water (2X), brine (1X) and then dried over MgSO4 . The compound is purified by flash column chromatography to yield the desired benzylidene.
The benzylidene is then azeotroped with benzene (2X 100 mL) and then dried overnight under vacuum over P2O 5. A mixture of benzylidene, dibutyl tin oxide ( 1.2 equiv.) and dry methanol (.25 M) are heated at reflux for 4 h until the solution became clear and homogeneous. (An automatic stirring apparatus may be necessary.) The solvent is next removed in vacuo to give a foamy white tin complex which was then azeotroped with benzene (2X) and dried (2 h to overnight) under vacuum over P2O 5. Next, anhydrous DMF (.2M) is added to redissolve the tin complex and then CsF (1.2 equiv.) and finally Benzyl bromide (1.5 equiv.) are added and then heated (40 °C) overnight. The clear solution is partially distilled under vacuum, (3.3 mm Hg, 75- 100 °C) to obtain 1/5 the original volume of solvent. Reaction mixture was then diluted with ethyl acetate (2L) and washed with a small amount of water (2X) to remove cesium salts. Aqueous layer is back extracted with ethyl acetate (3X) and then recombined with the organic layer which was then dried over MgSO4 and evaporated. Purification by flash column chromatography yields the desired benzyl ether 2000 . F o r related chemistry see Nagashima, N.; Ohno, M. Chemistry Letters, Chem. Soc. of Japan 1987, 141.
Preparation of compound 2010.
Procedure adopted from Johansson R.; Samuelsson; B. J. Chem. Soc , Chem. Commun. , 1 984 , 201. To a solution of the benzylidene acetal (1 equiv) and sodium cyanoborohydride (5 equiv.) in DMF (.125 M) containing powedered 3 angtrsom molecular sieves is added trifluoroacetic acid (10 equiv) and the reaction is allowed to stir at 0 °C until no starting material remains. Reaction mixture is then diluted with ethyl acetate (2L) and washed with a small amount of water (2X) and brine (2X). Aqueous layer is back extracted with ethyl acetate (3X) and then recombined with the organic layer which was then dried over MgSO4 and evaporated. Purification by flash column chromatography yields the desired benzyl ether 2010. Preparation of compound 2020.
To a solution of 2010 ( 1.0 equiv) was added methylene chloride (.1 M) and benzaldehyde ( 1.1 equiv), and the solution was exposed to ZnCI (1.1 equiv) at 25 °C and allowed to stir for 2.5 hour. The solution is then diluted with ether and then washed with a saturated solution of sodium bicarbonate (2X), water (2X), brine (1X) and then dried over MgSO4 . The compound is purified by flash column chromatography to yield the desired benzylidene 2020.
Preparation of compound 2030.
To a solution of alcohol 2020 (22.0 g, .1068 mol, 1.0 equiv.) in THF (0.5 M) at 0 °C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction iss cooled to 0 °C and treated with benzyl bromide (1.0 equiv.) and stirred for 1.5 h. A saturated solution of ammonium chloride (50 mL) is added dropwise to quench the reaction mixture at 0 °C and the mixture was diluted with ethyl acetate, washed with water (2X), brine (1X), dried over MgSO4 and evaporated. Purification by flash column chromatography yields tribenzyl ether 2030.
Preparation of compound 2040.
Procedure as adopted from Hanessian S.; Organic Syntheses
1987 , 243. To a suspension containing 1.0 equivalent of benzylidene 2 030 in one molar carbon tetrachloride and 1 , 1 ,2,2-tetrachloroethane ( 1.5 equivalent) is added 1 .2 equivalents of N-bromosuccinimide and 0.5 equivalents of barium carbonate. The resulting suspension is heated at the reflux temperature of the mixture with mechanical stirring for a period of 2.5 hour and filtered while hot. The solution is washed with water (3X), then dried over anhydrous sodium sulfate and evaporated. Purification by flash column chromatography yields
tribenzyl ether 2040.
Preparation of compound 2050.
To a solution of 2040 ( 1.0 equivalents) in methylene chloride (.10 M), is added diisopropylethylamine (4.0 equivalents) at 25 °C. The reaction is stirred for 5 minutes and then 2-cyanoethyl- N, N-diisopropyl-chlorophosphoramidite ( 1.5 equiv) is added, as prepared from the procedures of Sinha et al. Nucl. Acids Res. 1984, 12, 4539. After 15 minutes the reaction is
brine (1X) and is then dried (MgSO4 ) and concentrated. Purification by flash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 2050 (as shown in scheme 2000).
Preparation of compound 2060
To a solution of alcohol 2040 (1.0 equiv.) in THF (0.5 M) at 0 °C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction is cooled to 0 °C and exposed to the solid support functionalized with a bromide linker or any reasonable leaving group attached (1.0 equiv.) and stirred for 2 hours. A saturated solution of ammonium chloride (50 mL) is added dropwise to quench the reaction mixture at 0 °C and the support was washed with ethyl acetate, 1 % NaOH in methanol (2X) to remove the benzoate and finally brine (1X) to give 2 0 6 0 . The solid support used is the standard N-(2- Amino ethyl)- 3 -amino -propyl glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
To a solution of 76 ( 1.0 equiv) was added methylene chloride (.1 M) and benzaldehyde ( 1.1 equiv), and the solution was exposed to ZnCI (1.1 equiv) at 25 °C and allowed to stir for 2.5 hour. The solution is then diluted with ether and then washed with a saturated solution of sodium bicarbonate (2X), water (2X), brine (1X) and then dried over MgSO4 . The compound is purified by flash column chromatography to yield the desired benzylidene.
Procedure adopted from Johansson R.; Samuelsson; B. J. Chem. Soc , Chem. Commun. , 1 984 , 201. To a solution of the benzylidene acetal ( 1 equiv) and sodium cyanoborohydride (5 equiv.) in DMF (.125 M) containing powedered 3 angtrsom molecular sieves is added trifluoroacetic acid (10 equiv) and the reaction is allowed to stir at 0 °C until no starting material remains. Reaction mixture is then diluted with ethyl acetate
(2L) and washed with a small amount of water (2X) and brine (2X). Aqueous layer is back extracted with ethyl acetate (3X) and then recombined with the organic layer which was then dried over MgSO4 and evaporated. Purification by flash column chromatography yields the desired benzyl ether 2070.
Preparation of compound 2080
To a solution of 2070 ( 1.0 equivalents) in methylene chloride (.10 Molar), is added triethylamine ( 1.1 equivalents) at 0 °C.
Subsequent addition of tertbutyldiphenylsilylchloride ( 1.1 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4) and concentrated. Purification by flash column chromatography affords the TBDPS ether which is subsequently carried on as follows:
The TBDPS ether is then azeotroped with benzene (2X 100 mL) and then dried overnight under vacuum over P2O 5 . A mixture of benzylidene, dibutyl tin oxide (1.2 equiv.) and dry methanol
(.25 M) are heated at reflux for 4 h until the solution became clear and homogeneous. (An automatic stirring apparatus may be necessary.) The solvent is next removed in vacuo to give a foamy white tin complex which was then azeotroped with benzene (2X) and dried (2 h to overnight) under vacuum over P2O 5. Next, anhydrous DMF (.2M) is added to redissolve the tin complex and then CsF (1.2 equiv.) and finally Benzoyl bromide for the benzoate formation, ( 1.5 equiv.) are added and then heated (40 °C) overnight. The clear solution is partially distilled under vacuum, (3.3 mm Hg, 75- 100 °C) to obtain 1/5 the original volume of solvent. Reaction mixture was then diluted with ethyl acetate (2L) and washed with a small amount of water (2X) to remove cesium salts. Aqueous layer is back extracted with ethyl acetate (3X) and then recombined with the
organic layer which was then dried over MgS04 and evaporated. Purification by flash column chromatography yields the desired benzyl ether 2080. For related chemistry see Nagashima, N.; Ohno, M. Chemistry Letters, Chem. Soc. of Japan 1987, 141.
Preparation of compound 2090
To a solution of alcohol 2080 (1.0 equiv.) in THF (0.5 M) at 0 °C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction is cooled to 0 °C and treated with benzyl bromide (1.0 equiv.) and stirred for 1.5 h. The compound is then treated with tetrabutylammonium fluoride (2.0 equivalents) and allowed to stir for an additional 2 hours. A saturated solution of ammonium chloride (50 mL) is then added dropwise to quench the reaction mixture at 0 °C and the mixture was diluted with ethyl acetate, washed with water (2X), brine (1X), dried over MgSO4 and evaporated. Purification by flash column chromatography yields tribenzyl ether 2090. Preparation of compound 2100
To a solution of 2090 ( 1.0 equivalents) in methylene chloride (.10 M), is added diisopropylethylamine (4.0 equivalents) at 25 °C. The reaction is stirred for 5 minutes and then 2-cyanoethyl- N, N-diisopropyl-chlorophosphoramidite ( 1.5 equiv) is added, as
prepared from the procedures of Sinha et al. Nucl. Acids Res. 1984, 12, 4539. After 15 minutes the reaction is complete and is next diluted with ether and next washed with brine (1X) and is then dried (MgSO4) and concentrated. Purification by flash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 2100 (as shown in scheme 2002).
To a solution of alcohol 2090 (1.0 equiv.) in THF (0.5 M) at 0 °C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction is cooled to 0 °C and exposed to the solid support functionalized with a bromide linker or any reasonable leaving group attached ( 1.0 equiv.) and stirred for 2 hours. A saturated solution of ammonium chloride (50 mL) is added dropwise to quench the reaction mixture at 0
°C and the support was washed with ethyl acetate, 1 % NaOH in methanol (2X) to remove the benzoate and finally brine (1X) to give 2 1 1 0 . The solid support used is the standard N-(2- Aminoeteyl)-3-amino-propyI glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
Preparation of compound 2120
To a solution of 76 ( 1.0 equiv) was added methylene chloride (.1 M) and benzaldehyde ( 1.1 equiv), and the solution was exposed to ZnCI (1.1 equiv) at 25 °C and allowed to stir for 2.5
hour. The solution is then diluted with ether and then washed with a saturated solution of sodium bicarbonate (2X), water (2X), brine (1X) and then dried over MgSO4 . The compound is purified by flash column chromatography to yield the desired
benzylidene and carried on as follows:
To a solution of benzylidene (1.0 equiv.) in THF (0.5 M) at 0 °C, is added NaH ( 1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction is cooled to 0 °C and treated with benzyl bromide ( 1.0 equiv.) and stirred for 1.5 h. A saturated solution of ammonium chloride (50 mL) is then added dropwise to quench the reaction mixture at 0 °C and the mixture was diluted with ethyl acetate, washed with water (2X), brine (1X), dried over MgSO4 and evaporated. Purification by flash column chromatography yields tribenzyl ether 2120.
Preparation of compound 2130
Procedure adopted from Johansson R.; Samuelsson; B. J. Chem. Soc , Chem. Commun. , 1984 , 201. To a solution of the benzylidene acetal 2120 ( 1 equi v) and sodium cyanoborohydride (5 equiv.) in DMF (.125 M) containing powedered 3 angtrsom molecular sieves is added trifluoroacetic acid ( 10 equiv) and the reaction is allowed to stir at 0 °C until no starting material remains. Reaction mixture is then diluted with ethyl acetate (2L) and washed with a small amount of water (2X) and brine (2X). Aqueous layer is back extracted with ethyl acetate (3X) and then recombined with the organic layer which was then dried over MgSO4 and evaporated. Purification by
flash column chromatography yields the desired benzyl ether 2130.
Preparation of compound 2140
To a solution of 2130 ( 1.0 equivalents) in methylene chloride
(.10 Molar), is added triethylamine (1.1 equivalents) at 0 °C. Subsequent addition of tertbutyldiphenylsilylchloride ( 1.1 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried (MgSO4 and concentrated. Purification by flash column chromatography affords the TBDPS ether which is subsequently carried on as follows:
To a solution of TBDPS ether (1.0 equiv.) in THF (0.5 M) at 0 °C, is added NaH (1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction is cooled to 0 °C and treated with benzoyl bromide to afford benzoate formation ( 1.0 equiv.) and stirred for 1.5 h. A saturated solution of ammonium chloride (50 mL) is then added dropwise to quench the reaction mixture at 0 °C and the mixture was diluted with ethyl acetate, washed with water (2X), brine (1X), dried overMgSO4 and evaporated. Purification by flash column chromatography yields tribenzyl ether 2140.
Preparation of compound 2150
The compound 2140 is then treated with tetrabutylammonium fluoride (2.0 equivalents) in THF (.1 Molar) and allowed to stir for an additional 2 hours at 25 °C. A saturated solution of ammonium chloride (50 mL) is then added dropwise to quench the reaction mixture at 0 °C and the mixture was diluted with ethyl acetate, washed with water (2X), brine (1X), dried overMgSO4 and evaporated. Purification by flash column chromatography yields tribenzyl ether 2150.
Preparation of compound 2160
To a solution of 2150 (1.0 equivalents) in methylene chloride (.10 M), is added diisopropylethylamine (4.0 equivalents) at 25 °C. The reaction is stirred for 5 minutes and then 2-cyanoethyl-
N, N-diisopropyl-chlorophosphoramidite ( 1.5 equiv) is added, as prepared from the procedures of Sinha et al. Nucl. Acids Res. 1984 , 12, 4539. After 15 minutes the reaction is complete and is next diluted with ether and next washed with brine (1X) and is then dried (MgSO4) and concentrated. Purification by flash column chromatography (silica, 30% ethyl acetate in petroleum ether) affords compound 2160 (as shown in scheme 2004).
Preparation of compound 2170
To a solution of alcohol 2150 (1.0 equiv.) in THF (0.5 M) at 0 °C, is added NaH ( 1.0 equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction is cooled to 0 °C and exposed to the solid support functionalized with a bromide linker or any reasonable leaving group attached (1.0 equiv.)
and stirred for 2 hours. A saturated solution of ammonium chloride (50 mL) is added dropwise to quench the reaction mixture at 0 °C and the support was washed with ethyl acetate, 1 % NaOH in methanol (2X) to remove the benzoate and finally brine (1X) to give 2170. The solid support used is the standard
N- (2-Aminoethyl)-3-amino-propyl glass support; amino- polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.).
Prepartion of compound 3010
Procedure as described in Methods in Carbohydrate chemistry, Whistler, R., II, 1963, p. 327. A mixture of 80 g anhydrous D- glucosamine hydrochloride or D-galactosamine hydrochloride from Aldrich chemical company, in 200 mL. methanol and 20g Dowex 50 (H+) acidic resin, is stirred at the boiling point in a round bottom flask. After 24-hr. reaction time, the resin is removed by filtration and ished three times with 20 ml. of methanol. The filrate and washings are combined and concentrated to about 125 ml by rotovap. The concentrate is allowed to cool to room temperature and the product crystallizes overnight and carried on as follows:
The methyl glycoside is dissolved in chloroform (.5 M) and to it, is added phthalic anhydride ( 1.5 equiv.) and the reaction mixture is allowed to reflux at 70 °C for 4 h. The product
3010 is then crystallized and carried onto the next step.
Preparation of compound 3020
To a solution of alcohol 3010 (1.0 equiv.) in THF (0.5 M) at 0 °C, is added NaH (3.3equiv., 35% dispersion in mineral oil) over several portions. The reaction mixture is warmed to room temperature and stirred lh. Next, the reaction is cooled to 0 °C and treated with benzyl bromide (3.3 equiv.) and stirred for 1.5 h. A saturated solution of ammonium chloride (50 mL) is then added dropwise to quench the reaction mixture at 0 °C and the mixture was diluted with ethyl acetate, washed with water (2X), brine (1X), dried over MgSO4 and evaporated. Purification by flash column chromatography yields tribenzyl ether and is carried on as follows:
To a solution of tribenzyl ether in nitromethane is added trimethylsilyl cyanide (3.0 equivalents) and then SnCU ( .02 equivalents). The mixture is stirred for one hour and then an aqueous solution of sodium acetate was added to hydrolyze the remaining trimethylsilyl cyanide. The mixture is evaporated and the remaining oil is resuspended in dichloromethane and washed with sodium acetate solution (1X), water (1X), brine (1X) and then dried over magnesium sulphate and concentrated. The crude solid is then recrystallized from methanol is next dissolved in ethanol (0.15 M) and then concentrated H2S O4 (0.01
equivalents-catalytic) is added. The reaction mixture is heated to 85 °C for eight hours. The solution is next concentrated in vacuo and purification by flash column chromatography affords compound 3020 scheme 3000.
Prepartion of compound 3030
To a solution of 3020 ( 1.0 equivalents) in methylene chloride (.10 Molar), is added potassium carbonate (2.0 equivalents) at 0 °C. Subsequent addition of 9-fluorenylmethyl chloroformate (FMOC-Cl, 1.2 equivalents) is followed by stirring for 2 hours and then the reaction is diluted with diethylether and washed with ammonium chloride (2X), brine (1X) and then dried ( MgSO4 ) and concentrated. Purification by flash column chromatography affords product which is carried on as follows:
To a solution of ester in ethanol (.13 Molar), is added sodium ethoxide (0.3 equivalents) and the reaction mixture is stirred for two hours at room temperature. The solution is then concentrated in vacuo and purification by flash column chromatography affords compound 3030 scheme 3000.
Preparation of compound 3040
To a stirred solution of the acid 3030 ( 1.0 equivalents) and the (1.1 equivalents) in dimethylformamide (.10 Molar) at 25 °C, is added 1 -hydroxybenzotriazole (HOBT; 1.1 equivalents). N e x t
dicyclohexylcarbodiimide ( 1.2 equivalents) is added and the reaction is stirred for 2 hours. The mixture is then exposed to the solid support and mixed for 24 hours. (The solid support used is the standard N-(2-Aminoethyl)-3-amino-propyl glass support; amino-polystyrene resin; aminopropyl glass; isothiocyanato glass and others as purchased from Sigma company. All supports may be with or without a linker extending from the amino group on the support (eg. succinate linkage, amide, ether, alkyl chain with terminal carbon activated as free alcohol, bromide etc.)). The mixture is then
diluted with ether, washed with aqueous NaHCO3 (2X), water (2X), and brine (2X). Next, the compound/support ( 1.0 equivalents) in dimethyl-formamide (.10 Molar) at 25 °C, is added piperidine (1.1 equivalents). The support is stirred or exposed for 1 hour and is then diluted with ether, and washed with aqueous CUSO4 (2X), water (2X), and brine (2X). The final step affords compound 3040.
Phosphoramidate 138 (2 diastereomers) : IR, (neat) cm-1: 3089, 2964, 2927, 2856, 2253, 1497, 1455, 1396, 1363, 1253, 1184, 1156, 1094, 1028, 978, 876, 836, 779, 735,
1H-NMR (400 MHz, C6D6) : δ 7.34 (m, 5 H, Ph), 7.14 (m, 10 H, Ph), 4.97 (m, 4 H, CH2Ph), 4.78 (m, 2 H, CH2Ph), 4.07-3.24 (m, 13 H, OCH, OCH2, CH2CN), 1.81 (m, 2 H, CH(CH3)2), 1.16 (m, 12 H, CH3CH), 1.03, 1.02 (2 s, 9 H, 1BuSi), 0.20, 0.18, 0.16, 0.15, (4 s, 6 H, Me2Si);
HRMS: C43H6OO7N2PSi, Calc . (M+Cs+) : 911.3197; found:
911.3185.
Naphthoylester 136 IR, (neat) cm-i. 3494, 3062, 2919, 1716, 1630, 1600, 1454, 1355, 1284, 1228, 1197, 1091,
779, 736; 1H-NMR (250 MHz, CDCI3): δ 8.58 (s, 1 H, Ar), 8.00 (m, 2 H, Ar), 7.89 (m, 2 H, Ar), 7.59 (m, 2 H, Ar), 7.32 (m, 15 H, Ph), 4.95 (m, 3 H), 4.90 (d, -7=4.5 Hz, 1 H), 4.69 (m, 3 H), 4.52 (dd, J = 3.9, 12.0 Hz, 1 H), 3.91 (dd J = 2.6, 12.0, 1 H), 3.83 (d, J =
8.3, 1 H), 3.70 (m, 4 H), 3.96 (m, 1 H), 2.25 (s, 1 H, OH). HRMS: C39H38O7 Calc. (M+Cs+) : 751.1672; found: 751.1668. Dimer 142 IR, (neat) cirri : 3397, 3030, 2923, 2254, 1718,
1653, 1629, 1497, 1453, 1355, 1284, 1227, 1197, 1094, 1029, 780. 1H-NMR (400 MHz, C6D6) : δ 8.82 (s, 1 H, Ar), 8.26 (d, 1 H, Ar), 7.72 (m, 1 H, Ar), 7.61 (m, 1 H, Ar), 7.48 (m, 1 H, Ar), 7.37-6.95 (m, 32 H, Ar, Ph), 4.89-4.18 (m, 21 H, CH2Ph, CH2-Ar, -CH2CH2CN, CHCH2-Ar and CH2OH), 3.95-3.45 (m, 13 H, CH- and CH2-sugar), 1.71 (s, 1 H, OH); HRMS: C170H72O5NP calc. (M+H+):
1198.4718; found: 1198.4715.
Tetramer 150 IR, (neat) cm-1: 3420, 3064, 2924, 2255, 1721, 1497, 1455, 1357, 1278, 1028, 737. 1H-NMR (400 MHz, CDCI3) : δ 8.41 (s, 1 H, Ar), 8.00 (m, 2 H, Ar), 7.91 (m, 2 H, Ar), 7.55 (m, 2 H, Ar), 7.30 (m, 60 H, Ph), 4.93-4.05 (m, 39 H, CH2Ph, CH2-Ar, CH2CH2CN and CH2OH), 3.88-3.27 (m, 23 H, CH- and CH2-sugar), 2.58 (s, 1 H, OH). HRMS: C132H140O31N3P3 Calc. (M+Cs+):
2488.7738; found: 2488.7758.
Tetramer 154 IR, (neat) cm-1 : 3376, 2934, 1450, 1244, 1110, 1088. 1H-NMR (400 MHz, D2O): δ 8.41 (s, 1 H, Ar), 8.00 (m, 2 H, Ar), 7.91 (m, 2 H, Ar), 7.55 (m, 2 H, Ar), 4.93-4.05 (m, 4 H, CH2-Ar and CH2OH), 3.88-3.27 (m, 32 H, CH- and CH2-sugar); HRMS: C39H59O31P3 Calc.
(M+H+) : 1117.2331; found: 1117.2350.
Claims
1. An oligomeric carbopeptoid compound comprising carbohydrate amino acid subunits (CA's) coupled to one another via an amide linkage having a carbonyl carbon and an amido nitrogen represented by the following formula:
CAι-(CO-NH)-CA2
wherein:
CA1 is a first carbohydrate amino acid subunit having an anomeric carbon bonded to the carbonyl carbon of said amide linkage for forming a C-glycosidic linkage therewith and
CA2 is a second carbohydrate amino acid subunit having a non-anomeric carbon bonded to the amido nitrogen of said amide linkage.
2. In a process for synthesizing an oligomeric carbopeptoid compound, a coupling step wherein two or more carbohydrate amino acid subunits (CA's) are coupled by means of an amide linkage having a carbonyl carbon and an amido nitrogen for synthesizing said oligomeric carbopeptoid compound, said amide linkage being represented by a formula as follows:
CA1-(CO-NH)-CA2
wherein:
CA1 is a first carbohydrate amino acid subunit having an anomeric carbon bonded to the carbonyl carbon of said amide linkage for forming a C-glycosidic linkage therewith; and
CA2 is a second carbohydrate amino acid subunit having a non-anomeric carbon bonded to the amido nitrogen of said amide linkage.
3 . A library of oligomeric carbopeptoid compounds employable f or drug screening , each oligomeric carbopeptoid compound inc luding at leas t two carbohydrate amino acid subunits (CA ' s ) coupled to one another via an amide linkage having a carbonyl carbon and an amido nitrogen , said amide linkage being represented by the following formula :
CA1- (CO-NH) -CA2
wherein:
CA1 is a first carbohydrate amino acid subunit having an anomeric carbon bonded to the carbonyl carbon of said amide linkage for forming a C-glycosidic linkage therewith; and
CA2 is a second carbohydrate amino acid subunit having a non-anomeric carbon bonded to the amido nitrogen of said amide linkage.
4. An improved process for synthesizing a library of oligomers, the process employing an elongation step wherein subunits are coupled to one another to produce the oligomers, wherein the improvement comprises:
in said elongation step the oligomer includes at least two carbohydrate amino acid subunits (CA's) coupled to one another via an amide linkage having a carbonyl carbon and an amido nitrogen represented by the following formula:
CA1-(CO-NH)-CA2
wherein:
CA1 is a first carbohydrate amino acid subunit having an anomeric carbon bonded to the carbonyl carbon of said amide linkage for forming a C-glycosidic linkage therewith; and CA2 is a second carbohydrate amino acid subunit having a non-anomeric carbon bonded to the amido nitrogen of said amide linkage.
5. A derived carbohydrate amino acid having an anomeric carbon and non-anomeric carbons,
said anomeric carbon being substituted with a carboxyl radical,
each of said non-anomeric carbons being substituted with a radical selected from the group consisting of blocked hydroxyl, blocked amino, differentially protected amino, and hydrogen, with the proviso that at least one radical is a differentially protected amino.
6. A derived carbohydrate amino acid having an anomeric carbon and non-anomeric carbons,
said anomeric carbon being substituted with a carboxyl radical,
each of said non-anomeric carbons being substituted with a radical selected from the group consisting of blocked hydroxyl, blocked amino, unprotected amino, and hydrogen, with the proviso that at least one radical is an unprotected amino and at least one radical is a blocked hydroxyl or amino.
7. An oligomeric carbonucleotoid molecule comprising carbohydrate C-glycoside subunits (CG's) coupled to one another via a phosphodiester linkage represented by the following structure:
CG1-C1'-(O-PO(OH)-O)-CG2 wherein:
(O-PO (OH)-O)) is said phosphodiester linkage; CG1-C1' is a first carbohydrate C-glycoside subunit having an anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1' being bonded to said phosphodiester linkage; and
CG2 is a second carbohydrate C-glycoside subunit having a non-anomeric carbon bonded to said phosphodiester linkage.
8. In a process for synthesizing an oligomeric carbonucleotoid molecule, a coupling step wherein two or more carbohydrate C-glycoside subunits (CG's) are coupled by means of a phosphodiester linkage, said phosphodiester linkage being represented by a formula as follows:
CG1-C1'-(O-PO(OH)-O)-CG2 wherein:
(O-PO(OH)-O) is said phosphodiester linkage;
CG1-C1' is a first carbohydrate C-glycoside subunit having an anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1' being bonded to said phosphodiester linkage; and
CG2 is a second carbohydrate C-glycoside subunit having a non-anomeric carbon bonded to said phosphodiester linkage.
9. A library of oligomeric carbonucleotoid molecules employable for drug screening, each oligomeric carbonucleotoid molecule including at least two carbohydrate
C-glycoside subunits (CG's) coupled to one another by means of a phosphodiester linkage, said phosphodiester linkage being represented by a formula as follows:
CG1-C1'1(O-PO(OH)-O)-CG2 wherein:
(O-PO(OH)-O) is said phosphodiester linkage;
CG1-C1' is a first carbohydrate C-glycoside subunit having an anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1' being bonded to said phosphodiester linkage; and
CG2 is a second carbohydrate C-glycoside subunit having a non-anomeric carbon bonded to said phosphodiester linkage.
10. An improved process for synthesizing a library of oligomers, the process employing an elongation step wherein subunits are coupled to one another to produce the oligomers, wherein the improvement comprises:
in said elongation step the oligomer is a carbonucleotoid including at least two carbohydrate C- glycoside subunits (CG's) are coupled by means of a phosphodiester linkage, said phosphodiester linkage being represented by a formula as follows:
CG1-C1'-(O-PO(OH)-O)-CG2 wherein:
(O-PO(OH)-O) is said phosphodiester linkage;
CG1-C1' is a first carbohydrate C-glycoside subunit having an anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1.' being bonded to said phosphodiester linkage; and
CG2 is a second carbohydrate C-glycoside subunit having a non-anomeric carbon bonded to said phosphodiester linkage.
11. A derived carbohydrate C-glycoside having an anomeric carbon and non-anomeric carbons,
said anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1' being bonded to an activated phosphite,
each of said non-anomeric carbons being substituted with a radical selected from the group consisting of blocked hydroxyl, differentially protected hydroxyl, and hydrogen, with the proviso that at least one radical is a differentially protected hydroxyl.
12. A derived carbohydrate C-glycoside having an anomeric carbon and non-anomeric carbons,
said anomeric carbon forming a C-glycosidic bond with a carbon C1', said carbon C1' being bonded to an activated phosphite,
each of said non-anomeric carbons being substituted with a radical selected from the group consisting of blocked hydroxyl, unprotected hydroxyl, and hydrogen, with the proviso that at least one radical is an unprotected hydroxyl and at least one radical is a blocked hydroxyl.
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US08/913,035 US6204376B1 (en) | 1996-03-08 | 1996-03-08 | Carbopeptoids and carbonucleotoids |
EP96908737A EP0827406A1 (en) | 1995-03-08 | 1996-03-08 | Carbopeptoids and carbonucleotoids |
AU51882/96A AU717099B2 (en) | 1995-03-08 | 1996-03-08 | Carbopeptoids and carbonucleotoids |
US10/140,597 US20030013870A1 (en) | 1996-03-08 | 2002-05-07 | Carbopeptoids and carbonucleotoids |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
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US40103995A | 1995-03-08 | 1995-03-08 | |
US08/401,039 | 1995-03-08 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
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US09/417,877 Division US6384211B1 (en) | 1996-03-08 | 1999-10-13 | Carbopeptoids and carbo-nucleotoids |
Publications (2)
Publication Number | Publication Date |
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WO1996027379A1 WO1996027379A1 (en) | 1996-09-12 |
WO1996027379A9 true WO1996027379A9 (en) | 1996-11-28 |
Family
ID=23586019
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PCT/US1996/003227 WO1996027379A1 (en) | 1995-03-08 | 1996-03-08 | Carbopeptoids and carbonucleotoids |
Country Status (4)
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EP (1) | EP0827406A1 (en) |
AU (1) | AU717099B2 (en) |
CA (1) | CA2214789A1 (en) |
WO (1) | WO1996027379A1 (en) |
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ATE252914T1 (en) | 1996-08-13 | 2003-11-15 | Chiron Corp | POLYNUCLEOTIDE DELIVERY COMPOSITIONS |
US5756712A (en) * | 1997-01-23 | 1998-05-26 | E. I. Du Pont De Nemours And Company | Peptidodisaccharides as oligosaccharide mimetics |
US5919967A (en) * | 1997-04-11 | 1999-07-06 | Epix Medical, Inc. | Process for synthesizing phosphodiesters |
US6197332B1 (en) | 1997-08-13 | 2001-03-06 | Chiron Corporation | Lipid-conjugated polyamide compounds and related compositions and methods thereof |
CA3120451A1 (en) * | 2018-11-22 | 2020-05-28 | Idorsia Pharmaceuticals Ltd | Stable vaccine against clostridium difficile with modifications to the ps-ii saccharide |
CN110526950B (en) * | 2019-09-23 | 2023-08-04 | 济南山目生物医药科技有限公司 | Preparation method of alpha-five-O-acetyl mannose |
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US5212298A (en) * | 1989-08-16 | 1993-05-18 | Monsanto Company | Method for producing synthetic N-linked glycoconjugates |
US5573905A (en) * | 1992-03-30 | 1996-11-12 | The Scripps Research Institute | Encoded combinatorial chemical libraries |
AU7193494A (en) * | 1993-07-21 | 1995-02-20 | Oxford Glycosystems Ltd | Saccharides, their synthesis and use |
WO1995018971A1 (en) * | 1994-01-11 | 1995-07-13 | Affymax Technologies N.V. | Methods for the solid phase synthesis of glycoconjugates |
-
1996
- 1996-03-08 EP EP96908737A patent/EP0827406A1/en not_active Withdrawn
- 1996-03-08 CA CA 2214789 patent/CA2214789A1/en not_active Abandoned
- 1996-03-08 WO PCT/US1996/003227 patent/WO1996027379A1/en not_active Application Discontinuation
- 1996-03-08 AU AU51882/96A patent/AU717099B2/en not_active Ceased
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