EP0591256A1 - Methods for the synthesis of monofucosylated oligosaccharides terminating in di-n-acetyllactosaminyl structures - Google Patents
Methods for the synthesis of monofucosylated oligosaccharides terminating in di-n-acetyllactosaminyl structuresInfo
- Publication number
- EP0591256A1 EP0591256A1 EP92911443A EP92911443A EP0591256A1 EP 0591256 A1 EP0591256 A1 EP 0591256A1 EP 92911443 A EP92911443 A EP 92911443A EP 92911443 A EP92911443 A EP 92911443A EP 0591256 A1 EP0591256 A1 EP 0591256A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- group
- compound
- carbon atoms
- sialic acid
- hydrogen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Ceased
Links
- 238000000034 method Methods 0.000 title claims abstract description 72
- 238000003786 synthesis reaction Methods 0.000 title abstract description 49
- 230000015572 biosynthetic process Effects 0.000 title abstract description 46
- 229920001542 oligosaccharide Polymers 0.000 title description 38
- 150000002482 oligosaccharides Chemical class 0.000 title description 14
- 150000001875 compounds Chemical class 0.000 claims abstract description 85
- 238000002360 preparation method Methods 0.000 claims abstract description 24
- 108010001671 galactoside 3-fucosyltransferase Proteins 0.000 claims abstract description 14
- 229930182470 glycoside Natural products 0.000 claims description 106
- -1 L-fucosyl Chemical group 0.000 claims description 79
- 150000002338 glycosides Chemical class 0.000 claims description 54
- SQVRNKJHWKZAKO-OQPLDHBCSA-N sialic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)OC1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-OQPLDHBCSA-N 0.000 claims description 50
- 125000004432 carbon atom Chemical group C* 0.000 claims description 43
- 108090000141 Sialyltransferases Proteins 0.000 claims description 38
- 102000003838 Sialyltransferases Human genes 0.000 claims description 37
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical group N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 36
- SQVRNKJHWKZAKO-UHFFFAOYSA-N beta-N-Acetyl-D-neuraminic acid Natural products CC(=O)NC1C(O)CC(O)(C(O)=O)OC1C(O)C(O)CO SQVRNKJHWKZAKO-UHFFFAOYSA-N 0.000 claims description 35
- 239000000427 antigen Substances 0.000 claims description 34
- 102000036639 antigens Human genes 0.000 claims description 34
- 108091007433 antigens Proteins 0.000 claims description 34
- 230000000903 blocking effect Effects 0.000 claims description 31
- 229910052739 hydrogen Inorganic materials 0.000 claims description 30
- 239000001257 hydrogen Substances 0.000 claims description 30
- 150000002431 hydrogen Chemical group 0.000 claims description 22
- DKVBOUDTNWVDEP-NJCHZNEYSA-N teicoplanin aglycone Chemical group N([C@H](C(N[C@@H](C1=CC(O)=CC(O)=C1C=1C(O)=CC=C2C=1)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)OC=1C=C3C=C(C=1O)OC1=CC=C(C=C1Cl)C[C@H](C(=O)N1)NC([C@H](N)C=4C=C(O5)C(O)=CC=4)=O)C(=O)[C@@H]2NC(=O)[C@@H]3NC(=O)[C@@H]1C1=CC5=CC(O)=C1 DKVBOUDTNWVDEP-NJCHZNEYSA-N 0.000 claims description 22
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical group C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 claims description 21
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 claims description 20
- 241000124008 Mammalia Species 0.000 claims description 19
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 19
- 229910052757 nitrogen Inorganic materials 0.000 claims description 19
- 230000021633 leukocyte mediated immunity Effects 0.000 claims description 18
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 16
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 claims description 15
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical group [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims description 15
- 239000005864 Sulphur Chemical group 0.000 claims description 14
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical group [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 14
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 14
- 229910052760 oxygen Inorganic materials 0.000 claims description 14
- 239000001301 oxygen Chemical group 0.000 claims description 14
- SHZGCJCMOBCMKK-PQMKYFCFSA-N L-Fucose Natural products C[C@H]1O[C@H](O)[C@@H](O)[C@@H](O)[C@@H]1O SHZGCJCMOBCMKK-PQMKYFCFSA-N 0.000 claims description 13
- 125000000217 alkyl group Chemical group 0.000 claims description 13
- 230000028993 immune response Effects 0.000 claims description 13
- 230000004044 response Effects 0.000 claims description 13
- 230000001629 suppression Effects 0.000 claims description 12
- 125000005629 sialic acid group Chemical group 0.000 claims description 11
- 125000001424 substituent group Chemical group 0.000 claims description 11
- 150000001412 amines Chemical group 0.000 claims description 9
- 229910052799 carbon Inorganic materials 0.000 claims description 9
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 8
- 125000001475 halogen functional group Chemical group 0.000 claims description 8
- 125000003342 alkenyl group Chemical group 0.000 claims description 7
- 125000002947 alkylene group Chemical group 0.000 claims description 7
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 7
- 230000028709 inflammatory response Effects 0.000 claims description 7
- 125000006239 protecting group Chemical group 0.000 claims description 7
- 125000003545 alkoxy group Chemical group 0.000 claims description 6
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 claims description 6
- 239000008194 pharmaceutical composition Substances 0.000 claims description 5
- 150000003839 salts Chemical class 0.000 claims description 5
- 125000000717 hydrazino group Chemical group [H]N([*])N([H])[H] 0.000 claims description 2
- 238000007911 parenteral administration Methods 0.000 claims description 2
- 230000006698 induction Effects 0.000 claims 1
- 210000004698 lymphocyte Anatomy 0.000 claims 1
- WQZGKKKJIJFFOK-FPRJBGLDSA-N beta-D-galactose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-FPRJBGLDSA-N 0.000 abstract description 66
- 108010005774 beta-Galactosidase Proteins 0.000 abstract description 60
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine 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 abstract description 50
- OVRNDRQMDRJTHS-RTRLPJTCSA-N N-acetyl-D-glucosamine Chemical group CC(=O)N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-RTRLPJTCSA-N 0.000 abstract description 32
- 230000009450 sialylation Effects 0.000 abstract description 27
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 62
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 42
- 239000000203 mixture Substances 0.000 description 39
- 108010019236 Fucosyltransferases Proteins 0.000 description 31
- 102000006471 Fucosyltransferases Human genes 0.000 description 31
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 27
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 27
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 description 25
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 description 25
- 230000002255 enzymatic effect Effects 0.000 description 25
- 239000002904 solvent Substances 0.000 description 24
- 239000000047 product Substances 0.000 description 23
- 239000000243 solution Substances 0.000 description 23
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 description 22
- 230000033581 fucosylation Effects 0.000 description 21
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 20
- 238000012546 transfer Methods 0.000 description 19
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 18
- 150000001720 carbohydrates Chemical class 0.000 description 18
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical group OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 description 17
- 239000000370 acceptor Substances 0.000 description 16
- 235000000346 sugar Nutrition 0.000 description 16
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 15
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical class CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 15
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 12
- 102000004190 Enzymes Human genes 0.000 description 12
- 108090000790 Enzymes Proteins 0.000 description 12
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- LQEBEXMHBLQMDB-UHFFFAOYSA-N GDP-L-fucose Natural products OC1C(O)C(O)C(C)OC1OP(O)(=O)OP(O)(=O)OCC1C(O)C(O)C(N2C3=C(C(N=C(N)N3)=O)N=C2)O1 LQEBEXMHBLQMDB-UHFFFAOYSA-N 0.000 description 12
- 241000699670 Mus sp. Species 0.000 description 12
- 239000000126 substance Substances 0.000 description 12
- 208000027930 type IV hypersensitivity disease Diseases 0.000 description 12
- MIKUYHXYGGJMLM-GIMIYPNGSA-N Crotonoside Natural products C1=NC2=C(N)NC(=O)N=C2N1[C@H]1O[C@@H](CO)[C@H](O)[C@@H]1O MIKUYHXYGGJMLM-GIMIYPNGSA-N 0.000 description 11
- NYHBQMYGNKIUIF-UHFFFAOYSA-N D-guanosine Natural products C1=2NC(N)=NC(=O)C=2N=CN1C1OC(CO)C(O)C1O NYHBQMYGNKIUIF-UHFFFAOYSA-N 0.000 description 11
- LQEBEXMHBLQMDB-JGQUBWHWSA-N GDP-beta-L-fucose Chemical compound O[C@H]1[C@H](O)[C@H](O)[C@H](C)O[C@@H]1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C3=C(C(NC(N)=N3)=O)N=C2)O1 LQEBEXMHBLQMDB-JGQUBWHWSA-N 0.000 description 11
- 229910019142 PO4 Inorganic materials 0.000 description 11
- 210000004027 cell Anatomy 0.000 description 11
- 229940029575 guanosine Drugs 0.000 description 11
- 239000010452 phosphate Substances 0.000 description 11
- 125000005630 sialyl group Chemical group 0.000 description 11
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 10
- 102000004357 Transferases Human genes 0.000 description 10
- 108090000992 Transferases Proteins 0.000 description 10
- 239000000872 buffer Substances 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 10
- 239000001177 diphosphate Substances 0.000 description 10
- 230000002757 inflammatory effect Effects 0.000 description 10
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 10
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 description 9
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 description 9
- 239000000386 donor Substances 0.000 description 9
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 description 9
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- 150000004044 tetrasaccharides Chemical class 0.000 description 9
- 238000005406 washing Methods 0.000 description 9
- KBGAYAKRZNYFFG-BOHATCBPSA-N aceneuramic acid Chemical compound OC(=O)C(=O)C[C@H](O)[C@@H](NC(=O)C)[C@@H](O)[C@H](O)[C@H](O)CO KBGAYAKRZNYFFG-BOHATCBPSA-N 0.000 description 8
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- 235000020256 human milk Nutrition 0.000 description 8
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- 125000000325 methylidene group Chemical group [H]C([H])=* 0.000 description 8
- 238000000746 purification Methods 0.000 description 8
- IHQKEDIOMGYHEB-UHFFFAOYSA-M sodium dimethylarsinate Chemical compound [Na+].C[As](C)([O-])=O IHQKEDIOMGYHEB-UHFFFAOYSA-M 0.000 description 8
- 150000004043 trisaccharides Chemical class 0.000 description 8
- 102000015689 E-Selectin Human genes 0.000 description 7
- 108010024212 E-Selectin Proteins 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 7
- 108060003306 Galactosyltransferase Proteins 0.000 description 7
- 102000030902 Galactosyltransferase Human genes 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 7
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- 238000002955 isolation Methods 0.000 description 7
- 210000004185 liver Anatomy 0.000 description 7
- IWZSHWBGHQBIML-ZGGLMWTQSA-N (3S,8S,10R,13S,14S,17S)-17-isoquinolin-7-yl-N,N,10,13-tetramethyl-2,3,4,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthren-3-amine Chemical compound CN(C)[C@H]1CC[C@]2(C)C3CC[C@@]4(C)[C@@H](CC[C@@H]4c4ccc5ccncc5c4)[C@@H]3CC=C2C1 IWZSHWBGHQBIML-ZGGLMWTQSA-N 0.000 description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- TXCIAUNLDRJGJZ-BILDWYJOSA-N CMP-N-acetyl-beta-neuraminic acid Chemical compound O1[C@@H]([C@H](O)[C@H](O)CO)[C@H](NC(=O)C)[C@@H](O)C[C@]1(C(O)=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C(N=C(N)C=C2)=O)O1 TXCIAUNLDRJGJZ-BILDWYJOSA-N 0.000 description 6
- KFEUJDWYNGMDBV-LODBTCKLSA-N N-acetyllactosamine Chemical compound O[C@@H]1[C@@H](NC(=O)C)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 KFEUJDWYNGMDBV-LODBTCKLSA-N 0.000 description 6
- 206010070834 Sensitisation Diseases 0.000 description 6
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- 238000006911 enzymatic reaction Methods 0.000 description 6
- 210000000987 immune system Anatomy 0.000 description 6
- 230000001404 mediated effect Effects 0.000 description 6
- 150000004702 methyl esters Chemical class 0.000 description 6
- 239000011347 resin Substances 0.000 description 6
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- 230000008313 sensitization Effects 0.000 description 6
- 238000001228 spectrum Methods 0.000 description 6
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 5
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- FTVLMFQEYACZNP-UHFFFAOYSA-N trimethylsilyl trifluoromethanesulfonate Chemical compound C[Si](C)(C)OS(=O)(=O)C(F)(F)F FTVLMFQEYACZNP-UHFFFAOYSA-N 0.000 description 5
- GHYOCDFICYLMRF-UTIIJYGPSA-N (2S,3R)-N-[(2S)-3-(cyclopenten-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-hydroxy-3-(4-methoxyphenyl)-2-[[(2S)-2-[(2-morpholin-4-ylacetyl)amino]propanoyl]amino]propanamide Chemical compound C1(=CCCC1)C[C@@H](C(=O)[C@@]1(OC1)C)NC([C@H]([C@@H](C1=CC=C(C=C1)OC)O)NC([C@H](C)NC(CN1CCOCC1)=O)=O)=O GHYOCDFICYLMRF-UTIIJYGPSA-N 0.000 description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- 150000008481 L-fucoses Chemical class 0.000 description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 4
- HESSGHHCXGBPAJ-UHFFFAOYSA-N N-acetyllactosamine Natural products CC(=O)NC(C=O)C(O)C(C(O)CO)OC1OC(CO)C(O)C(O)C1O HESSGHHCXGBPAJ-UHFFFAOYSA-N 0.000 description 4
- 229930182475 S-glycoside Natural products 0.000 description 4
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/02—Acyclic radicals, not substituted by cyclic structures
- C07H15/04—Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/06—Pyrimidine radicals
- C07H19/10—Pyrimidine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
- C07H19/20—Purine radicals with the saccharide radical esterified by phosphoric or polyphosphoric acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
- C07H3/06—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
Definitions
- This invention is directed to methods for the preparation of monofucosylated and sialylated derivatives of the compound ⁇ Gal(1-4) ⁇ GlcNAc-(1-3) ⁇ Gal(1-4) ⁇ GlcNAc-OR.
- the methods of this invention provide for a multi-step synthesis wherein selective monofucosylation is accomplished on the 3-hydroxy group on only one of the GlcNAc units found in the ⁇ Gal(1-4) ⁇ GlcNAc-(1-3) ⁇ Gal(1-4) ⁇ GlcNAc-OR compound.
- oligosaccharides such as sialylated and fucosylated structures are involved as ligands in cell adhesion phenomena. 1
- oligosaccharide glycosides relating to blood group determinant structures have been found to impart immunosuppressive and tolerogenic
- oligosaccharide glycosides relating to blood group determinant structures include the compound
- Ippolito et al. 22 further discloses that blood group determinant oligosaccharide glycosides having a sialic acid group (or an analogue thereof) at the non-reducing sugar terminus of the oligosaccharide glycosides and which are also monofucosylated
- immunosuppressive and tolerogenic properties e.g., sialyl Lewis x — Compound III in FIG. 12 of Ippolito et a l . 22 ).
- a synthetic approach employing enzymatic sialylation and fucosylation steps is particularly appropriate in order to provide an efficient route for the preparation of sialylated and
- ⁇ (2-3)sialyltransferase from porcine submaxillary gland have often been used for synthetic purposes. 3
- the former two sialyltransferases are useful in sialylating a terminal ⁇ Gal(1-4) ⁇ GlcNAc- group in an oligosaccharide glycoside.
- sialyltrans-ferase which has a wide acceptor specificity, is useful in sialylating a terminal ⁇ Gal(1-3) ⁇ GlcNAc-group in oligosaccharide glycosides based on the Lewis c (Type I) backbone 4 but, because of the low affinity of this enzyme for the Type II backbone, the synthesis of sialylated N-acetyllactosaminyl structures, such as those present in the sialyl Lewis x , 5 sialyl dimeric Lewis x6 or the corresponding internally monofucosylated derivati v e 7 , by use of this sialyltransferase is much more difficult. 4
- Type II terminal structures such as:
- glycoconjugates will then depend upon the particular ⁇ (1-3) fucosyl-transferase present" 11 .
- the present invention is based, in part, on the discovery of synthetic pathways which utilizes enzymatic fucosylation and sialylation steps and which result in the selective formation of
- This invention is directed, in part, to the discovery that fucosylation onto the 3-hydroxyl group of the GlcNAc saccharide in a ⁇ Gal(1-4) ⁇ GlcNAc disaccharide via an ⁇ (1 ⁇ 3) fucosyltransferase (e.g., ⁇ Gal(1-3/4) ⁇ GlcNAc ⁇ (1-3/4) fucosyltransferase) is dependent on the presence of a 6-hydroxyl group on the Gal saccharide and when this hydroxyl group is blocked by a removable blocking group, fucosylation on the neighboring GlcNAc group is prevented.
- the methods of this invention employs this characteristic of ⁇ (1-3)fucosyltransferases to provide for a means to selectively monofucosylate compound 1a which are used advantageously to prepare compounds 5a and 12.
- the present invention is directed to the discovery of enzymatic methods and chemical/enzymatic methods to prepare the compound ⁇ Neu5Ac(2-3) ⁇ Gal(1-4) ⁇ GlcNAc(1-3) ⁇ Gal(1-4)-[ ⁇ Fuc(1-3)] ⁇ GlcNAc-OR.
- the present invention is directed to a method for preparation of a compound of the formula I:
- R is an aglycon group having at least one carbon atom
- Y is L-fucose
- Z is sialic acid or an analogue of sialic acid, which method comprises the following steps:
- R is as defined above and X is a removable blocking group
- the sialylation of the oligosaccharide glycoside so as to form an ⁇ (2-3)sialyl residue at the non-reducing sugar terminus of the oligosaccharide glycoside is necessarily after removing the blocking group because sialylation with an ⁇ (2-3)sialyltransferase requires the presence of a free hydroxyl group at the 6-position of the terminal galactose residue on the oligosaccharide glycoside.
- the present invention is directed to a method for preparation of a compound of the formula IV:
- R is an aglycon group having at least one carbon atom
- Y' is L-fucose
- Z is sialic acid or an analogue of sialic acid, which method comprises the following steps:
- R is as defined above and X' is a removable blocking group
- Preferred removable blocking groups for use in the above described methods include sialic acid groups and benzyl groups and any other group that can be introduced either enzymatically or chemically on the precursor leading to II or V and later selectively enzymatically or chemically removed in mild conditions compatible with the nature of the product.
- the blocking group X' is not sialic acid because this compound would be difficult to synthesize.
- the present invention is directed to a compound of the formula VII:
- R is an aglycon having at least 1 carbon atom
- Y and Y' are selected from the group
- the aglycon moiety, R is selected from the group consisting of -(A)-Z' wherein A represents a bond, an alkylene group of from 2 to 10 carbon atoms, and a moiety of the form -(CH 2 -CR 2 G) n - wherein n is an integer equal to 1 to 5;
- R 2 is selected from the group consisting of hydrogen, methyl, or ethyl;
- G is selected from the group consisting of hydrogen, halogen, oxygen, sulphur, nitrogen, phenyl and phenyl substituted with 1 to 3 substituents selected from the group consisting of amine, hydroxyl, halo, alkyl of from 1 to 4 carbon atoms and alkoxy of from 1 to 4 carbon atoms; and
- Z' is selected from the group consisting of hydrogen, methyl
- R 4 independently alkyl of from 1 to 4 carbon atoms and R 4 is an alkenyl group of from 3 to 10 carbon atoms with the proviso that when A is a bond then Z' is not hydrogen.
- the aglycon group is a hydrophobic group.
- the present invention is directed to a pharmaceutical composition suitable for administration to a mammal (e.g., human) which comprises a pharmaceutically inert carrier and an effective amount of the compound of Formula I or IV to modulate a cell-mediated immune response in said mammal.
- a mammal e.g., human
- the present invention is directed to a method for modulating a cell-mediated immune response in a mammal which method comprises administering to said mammal an amount of a compound of Formula I or IV effective in modulating said immune response.
- FIG. 1 illustrates the synthetic pathway leading to Sialyl dimeric Lewis x and internally monofucosylated derivatives thereof.
- the nomenclature for compound 1a is ⁇ Gal(1-4) ⁇ GlcNAc(1- 3) ⁇ Gal(1-4) ⁇ GlcNAc-OR, sometimes called di-N- acetyllactosaminyl tetrasaccharide.
- the hexasaccharide moiety present in compounds 5a and 5b in FIG. 1 is sometimes called VIM-2 epitope or CD- 65 5 and 7a and 7b are called sialyl dimeric Lewis x .
- FIG. 2 illustrates the synthetic pathway leading to the externally monofucosylated
- FIGURE 3 illustrates an enzymatic pathway leading to monofucosylated and monosialylated compounds of Formula I.
- FIGURE 4 illustrates an alternative chemical synthesis of trisaccharide 19 which can then be used as per Figure 3 to prepare monofucosylated and monosialylated compounds of Formula I.
- FIGURE 5 illustrates that the enzymatic pathway set forth in Figure 3 can be used to extend the structure of the hexasaccharides of Formula I.
- the present invention is directed, in part, to the discovery that selective monofucosylation of compound la (i.e., ⁇ Gal(1-4) ⁇ GlcNAc(1-3) ⁇ Gal- (1-4) ⁇ GlcNAc-OR), can be achieved by appropriately blocking the 6-hydroxy group on the galactose unit adjacent to GlcNAc unit (in the non-reducing sugar direction) so as to prevent fucosylation of the GlcNAc unit.
- compound la i.e., ⁇ Gal(1-4) ⁇ GlcNAc(1-3) ⁇ Gal- (1-4) ⁇ GlcNAc-OR
- the present invention is also directed, in part, to the discovery that the preparation of compounds of Formula I can be achieved by a complete enzymatic process or by a chemo/enzymatic process.
- the present invention is still further directed to the discovery that the compounds of Formula I and II are useful for in vivo modulation of a cell mediated immune response in mammals, including humans.
- cell-mediated immune response to an antigen in a mammal refers to those mammalian immune responses which are mediated by cell-cell interactions. Included within this term are cell mediated inflammatory responses to an antigen such as DTH responses as well as cell-mediated
- the cell-mediated immune response is a leucocyte-mediated response.
- N-acetyllactosamine or “LacNAc” refers to the disaccharide ⁇ Gal(1 ⁇ 4) ⁇ GlcNAc which is represented by the formula:
- di-N-acetyllactosaminyl structures means that one N-acetyllactosamine unit is
- Neu5Ac and the naturally occurring analogues of Neu5Ac, including N-glycolyl neuraminic acid
- sialyltransferase A complete list of naturally occurring sialic acids known to date are provided by Schauer 23 .
- Naturally occurring sialic acids which are recognized by a particular sialyltransferase so as to bind to the enzyme and are then available for transfer to an appropriate acceptor oligosaccharide structure are said to be compatible with the
- sialyltransferase and are sometimes referred to herein as a "compatible naturally occurring sialic acid".
- analogue of sialic acid refers to analogues of naturally occurring structures of sialic acid including those wherein the sialic acid unit has been chemically modified so as to introduce and/or remove one or more functionalities from such structures. For example, such modification can result in the removal of an -OH functionality, the introduction of an amine functionality, the
- analogues of sialic acid include, by way of example, 9-azido-Neu5Ac, 9-amino-Neu5Ac, 9-deoxy-Neu5Ac, 9-fluoro-Neu5Ac, 9-bromo-Neu5Ac, 8-deoxy-Neu5Ac, 8-epi-Neu5Ac, 7-deoxy-Neu5Ac, 7-epi-Neu5Ac, 7,8-bis-epi- Neu5Ac, 4-O-methyl-Neu5Ac, 4-N-acetyl-Neu5Ac, 4,7-di-deoxy-Neu5Ac, 4-oxo-Neu5Ac, 3-hydroxy-Neu5Ac, 3-fluoro-Neu5Ac acid as well as the 6-thio analogues of Neu5Ac
- the nomenclature employed herein in describing analogues of sialic acid is as set forth by Reuter et al. 24
- sialyltransferases are designed to transfer or donate compatible naturally occurring sialic acids
- analogues of Neu5Ac are sometimes referred to herein as “artificial donors”
- the compatible naturally occurring sialic acids are sometimes referred to herein as the "natural
- sialyltransferase refers to those enzymes which transfer a compatible naturally occurring sialic acid, activated as its cytidine monophosphate (CMP) derivative, to the terminal galactose group present on natural acceptor
- ⁇ Gal(1-4) ⁇ GlcNAc and ⁇ Gal(1-3) ⁇ GlcNAc disaccharides and include enzymes produced from microorganisms genetically modified so as to incorporate and express all or part of the sialyltransferase gene obtained from another source, including mammalian sources.
- sialyltransferases comprise those that have been identified in the literature as leading to the following structures:
- Analogues of sialic acid which are recognized by a particular sialyltransferase so as to bind to the enzyme and are then available for transfer to an appropriate acceptor oligosaccharide structure are said to be compatible with the sialyltransferase and are sometimes referred to herein as a "compatible analogue of sialic acid". Because the transfer reaction employs a sialyltransferase, it goes without saying that an analogue of sialic acid employed in such a reaction must be a compatible analogue of sialic acid.
- CMP-nucleotide derivative of Neu5Ac refers to the compound:
- CMP-derivatives of analogues of sialic acid refer to those compounds having structures similar to that above with the exception that the Neu5Ac residue is replaced with an analogue of sialic acid.
- ⁇ (1-3)fucosyltransferase refers to any fucosyltransferase which transfers L-fucose and compatible analogues of L-fucose from GDP-fucose to the 3 hydroxy position of GlcNAc in a LacNAc group ( ⁇ Gal(1-4) ⁇ GlcNAc) in an oligosaccha-ride glycoside and which does not discriminate between ⁇ Gal(1- 4)) ⁇ GlcNAc groups in the oligo-saccharide glycoside.
- the particular ⁇ (1-3)fucosyl-transferase employed is compatible with the intended reaction. That is to say that the selected ⁇ (1-3) fucosyltransferase will bind to the oligosaccharide glycoside employed and transfer L-fucose to the 3 hydroxy position of
- Suitable fucosyltrans- ferases include the known ⁇ Gal(1 ⁇ 3/4) ⁇ GlcNAc
- Compatible analogues of L-fucose refer to naturally occurring and synthetic analogues of fucose including those where the fucose unit has been chemically modified so as to introduce and/or remove one or more functionalities from this
- such modification can result in the removal of an -OH functionality, the introduction of an amine functionality, the
- fucose Certain compatible analogues of fucose are known in the art and include, by way of example, 3-deoxy-fucose, arabinose, and the like. 18
- removable blocking group refers to any group which when bound to the 6-hydroxyl of the galactose unit in a ⁇ Gal(1-4) ⁇ GlcNAc group prevents fucosylation of the 3-hydroxyl of the GlcNAc by an ⁇ (1-3)fucosyltransferase and which group can be removed by conventional chemical or enzymatic steps to reestablish the 6-hydroxyl on the galactose unit.
- the particular removable blocking group employed is not critical and preferred removable blocking groups include Neu5Ac and benzyl substituents and any other group that can be introduced either enzymatically or chemically on the precursor leading to II or V and later selectively enzymatically or chemically removed in mild conditions compatible with the nature of the product.
- contemplated blocking group is ⁇ -galactose which can be removed enzymatically with an ⁇ -galactosidase.
- removable protecting group refers to any group which when bound to one or more hydroxyl groups of the galactose, N-acetylglucosamine, etc. which prevent reactions from occurring at these hydroxyl groups and which protecting group can be removed by conventional chemical or enzymatic steps to reestablish the hydroxyl group.
- the particular removable protecting group employed is not critical and preferred removable hydroxyl protecting groups include conventional substituents such as benzyl, acetyl, chloroacetyl, benzylidine, t-butyldiphenylsilyl and any other group that can be
- pharmaceutically acceptable salts includes the pharmaceutically acceptable addition salts of the compounds of Formula I derived from a variety of organic and inorganic counter salts well known in the art and include, by way of example only, sodium, potassium, calcium, magnesium,
- ammonium tetralkylammonium, and the like.
- aglycon refers to the R substituent on the hexasaccharide glycosides of formula I and IV.
- R is an aglycon having at least 1 carbon atom.
- the aglycon moiety, R is selected from the group consisting of.
- A represents a bond, an alkylene group of from 2 to 10 carbon atoms, and a moiety of the form -(CH 2 -CR 2 G) n - wherein n is an integer equal to 1 to 5;
- R 2 is selected from- the group consisting of hydrogen, methyl, or ethyl; and
- G is selected from the group consisting of hydrogen, halogen, oxygen, sulphur, nitrogen, phenyl and phenyl
- Z' is selected from the group consisting of hydrogen, methyl, phenyl, nitrophenyl, and, when G is not oxygen, sulphur or nitrogen and A is not a bond, then Z' is also selected from the group consisting of -OH, -SH, -NH 2 , -NHR 3 , -N(R 3 ) 2 , -C(O)OH, -C(O)OR 3 , -C(O)NH-NH 2 , -C(O)NH 2 , -C(O)NHR 3 , -C(O)N(R 3 ) 2 , and -OR 4 wherein each R 3 is
- R 4 independently alkyl of from 1 to 4 carbon atoms and R 4 is an alkenyl group of from 3 to 10 carbon atoms with the proviso that when A is a bond, Z' is not hydrogen.
- the aglycon is preferably selected from the group consisting of - (A) -Z' ' wherein A is selected from the group consisting of an alkylene group of from 2 to 10 carbon atoms and a moiety of the form -(CH 2 -CR 5 G) n - wherein n is an integer equal to 1 to 5; R 5 is selected from the group consisting of hydrogen, methyl, or ethyl; and G is selected from the group consisting of hydrogen, oxygen, sulphur, nitrogen, phenyl and phenyl
- Z' ' is selected from the group consisting of hydrogen and, when G is not oxygen, sulphur or nitrogen, then Z' ' is also selected from the group consisting of -OH, -SH, -NH 2 , -NHR 6 ,
- each R 6 is independently alkyl of from 1 to 4 carbon atoms and R 7 is an alkenyl group of from 3 to 10 carbon atoms with the proviso that when A is a bond, Z is not hydrogen.
- the -(A)-Z' ' group defines a group capable of being linked to a carrier or is capable of being derivatized to a group which is capable of being linked to a carrier. The choice of an appropriate carrier may be useful in enhancing immunogenic properties.
- the nitro group is reduced to an amino group which can be protected as N-trifluoro-acetamido.
- the trifluoroacetamido group is removed thereby
- a linking arm containing sulfur is disclosed by Dahmen et al. 51 .
- the linking arm is derived from a 2-bromoethyl group which, in a
- Rana et al. 52 discloses a 6-trifluoroacetamido- hexyl linking arm (-O-(CH 2 ) 6 -NHCOCF 3 ) in which the trifluoroacetamido protecting group can be removed unmasking the primary amino group used for coupling.
- linking arms include the 7-methoxycarbonyl-3,6,dioxaheptyl linking arm 53 (-OCH 2 -CH 2 ) 2 OCH 2 CO 2 CH 3 ; the
- allyl linking arms can be derivatized in the presence of 2-aminoethanethiol 57 to provide for a linking arm -OCH 2 CH 2 CH 2 SCH 2 CH 2 NH 2 .
- R group can be an additional saccharide or an
- oligosaccharide containing a linking arm at the reducing sugar terminus oligosaccharide containing a linking arm at the reducing sugar terminus.
- the carrier is generally a small molecular weight, non-immunogenic or antigenic carrier
- linking including the linking to a fluorescent label, a radioactive label, biotin, or a photolabile linking arm or a moiety to be targetted.
- oligosaccharides and glycoconjugates because the aglycon moiety (R) is not hydrogen, a protein, or a lipid capable of forming a micelle or other large aggregate structure.
- Tetrasaccharide glycosides II and V are readily prepared either by complete chemical synthesis or a chemical/enzymatic synthesis as described below.
- tetrasaccharide glycoside II and V can be prepared by chemically coupling the individual saccharide units. Such coupling can readily be prepared using a convergent synthesis, i.e.,
- tetrasaccharide glycosides can be conducted in a sequential synthesis starting with the saccharide unit at the reducing sugar terminus and sequentially adding another saccharide unit until tetrasaccharide glycosides II and V are prepared.
- the first step of the synthesis involves the addition of the aglycon moiety at the anomeric carbon atom of the reducing sugar unit.
- the sugar donor is then reacted under catalytic conditions (e.g., a soluble silver salt such as silver trifluoromethanesulfonate, a Lewis acid such as boron trifluoride etherate or trimethylsilyltrifluoromethanesulfonate, or
- thioglycoside promoters such as methyl
- deblocked i.e., the removable blocking group X or X' is removed
- oligosaccharide glycoside is sialylated by contacting the appropriate oligosaccharide glycoside with an ⁇ (2-3)sialyltransferase and a compatible CMP-derivative of a sialic acid or an analogue thereof under conditions wherein the sialic acid or the analogue thereof is transferred to the non- reducing sugar terminus of the oligosaccharide glycoside.
- Suitable conditions known in the art, include the addition of the appropriate
- oligosaccharide glycoside and of the CMP-derivative of the sialic acid in an appropriate buffer such as 0.1 M sodium cacodylate in appropriate conditions of pH and temperature such as at a pH of 6.5 to 7.5 and a temperature between 25 and 45oC, preferably 35-40°C for 12 hours to 4 days.
- an appropriate buffer such as 0.1 M sodium cacodylate
- pH and temperature such as at a pH of 6.5 to 7.5 and a temperature between 25 and 45oC, preferably 35-40°C for 12 hours to 4 days.
- sialylated oligosaccharide glycoside can be isolated and purified using conventional methodology
- an analogue of sialic acid when transferred to the oligosaccharide glycoside by the sialyltransferase, the analogue is sometimes referred to as an artificial donor and the
- oligosaccharide glycoside is sometimes referred to as an artificial acceptor.
- Sialylation methods employing an artificial donor and an artificial acceptor are described by Venot et al., U.S. Patent Application Serial No. 07/771,007 filed October 2, 1992 which application is incorporated herein by reference in its entirety. Similarly, sialylation methods employing an artificial donor and an artificial acceptor
- analogues of sialic acid require the prior synthesis (i.e., activation) of their nucleotide (CMP) derivatives.
- Activation of the analogues of sialic acid is usually done by using the enzyme CMP-sialic acid synthase which is readily available and the literature provides examples of the activation of various analogues of sialic acid.
- the present invention is based, in part, on the discovery that, in Figure l, sialylation of deblocked pentasaccharide glycoside III so as to form an ⁇ (2-3)sialyl residue at the non-reducing sugar terminus of this oligosaccharide glycoside is necessarily after removing the removable blocking group because sialylation with an ⁇ (2-3)sialyltransferase requires the presence of a free hydroxyl group at the 6-position of the terminal galactose residue on the deprotected pentasaccharide glycoside III.
- sialylation of the tetrasaccharide glycoside V so as to form an ⁇ (2-3)sialyl residue at the non-reducing sugar terminus of this oligosaccharide glycoside is necessarily before the fucosylation step because sialylation with an ⁇ (2-3)sialyltransferase will not proceed if there is an ⁇ -fucose linked (1-3) to the neighboring N-acetylglucosamine.
- oligosaccharide glycoside II or pentasaccharide glycoside derived by sialylating tetrasaccharide glycoside V or trisaccharide 19 are fucosylated by contacting the appropriate oligosaccharide glycoside with an ⁇ (1-3)fucosyltransferase and a compatible GDP-derivative of L-fucose or an analogue of L-fucose under conditions wherein the fucose is
- Suitable conditions include the addition of the ⁇ (1-3)fucosyltransferase to a
- GDP-fucose is prepared in the methods described by Jiang et al. 59
- oligosaccharide glycoside is treated under conditions sufficient to effect removal of the blocking group.
- the specific conditions depend on the blocking group employed and are well known in the art. For example, when a benzyl blocking group is employed, this group is readily removed by hydrogenation techniques known in the art.
- the blocking group is sialic acid, it is removed in the manner depicted in the Examples set forth herein below.
- Figure 1 illustrates the synthesis of hexasaccharide glycoside I (compound 5a and 5b) and heptasaccharide glycoside (compound 7a and 7b).
- the tetrasaccharide la was
- a suitable immobilized sialidase e.g., a sialidase from
- the 8-methoxycarbonyloctyl glycoside of the starting tetrasaccharide 1a and trisaccharide 9 was used with the intention of taking advantage of the hydrophobic properties of the aglycone for
- Figure 1 also illustrates that heptasaccharide 7b was obtained by sequential sialylation of la by the ⁇ Gal(1-3/4) ⁇ GlcNAc ⁇ (2-3)sialyltransferase, followed by difucosylation of the intermediate 6a by the ⁇ Gal(1-3/4) ⁇ GlcNAc ⁇ (1-3/4)fucosyltransferase from human milk. In the conditions used, only the difucosylated product was obtained.
- hexasaccharide IV (compound 12.).
- Alternative syntheses for hexasaccharide I are set forth below and generally involve a
- GlcNAc-OR to form ⁇ Gal(1-4) ⁇ GlcNAc-OR (LacNAc-OR).
- Suitable enzymes include the GlcNAc ⁇ 1-4
- galactosyltransferase which transfers galactose from uridine 5'(galactopyranosyl)-diphosphate (UDP-Gal) to the 4-position of GlcNAc ⁇ -OR, where R can be an aglycone or a saccharide.
- This transferase is a commercial and versatile enzyme and accepts some modifications in the sugar portion of the donor 47 and in the acceptor 48-49 .
- N-acetylglucosamine is then transferred to the 3-position of the terminal ⁇ -galactose of LacNAc-OR (N-acetyllactosamine-OR — ⁇ Gal(1-4) ⁇ GlcNAc-OR) to produce the ⁇ GlcNAc(1-3) ⁇ Gal(1-4) ⁇ GlcNAc-OR
- Transferases which transfer N-acetylglucosamine from uridine 5'-(N-acetylglucosamine)-diphosphate (UDP-GlcNAc) to the 3 position of the terminal ⁇ -galactose of a N-acetyllactosamine moiety) are present in a variety of sources such as human serum 36-40 , human urine 41 , Novikoff tumor cell ascites fluid 42,43 , mouse T-lymphoma cells 44 , human milk 45 and human colonic adenocarcinoma cells 46 .
- the acceptor specificity of the transferases obtained has been well characterized using synthetic oligosaccharides.
- This enzyme requires a terminal ⁇ Gal(1-4) ⁇ Glc(NAc)-OR unit, where R can be an aglycone or a saccharide moiety and Glc(NAc) can be either GlcNAc or Glc
- R can be an aglycone or a saccharide moiety
- Glc(NAc) can be either GlcNAc or Glc
- the enzyme does not transfer to the structure ⁇ Gal(1-4)[ ⁇ Fuc(1-3)] ⁇ GlcNAc (Lewis x ) in which the fucose is attached to the penultimate GlcNAc 36,43 .
- Enzymatic transfer of fucose to the acceptor 19 by the milk Gal(1-3/4) GlcNAc ⁇ (1-3/4)fucosyltransferase specifically occurs on the internal GlcNAc leading to compound 20.
- the backbone of the tetrasaccharide 20 is then extended by transfer of a galactose residue leading to compound 21 by the bovine GlcNAc ⁇ 1-4 galactosyltransferase.
- Neu5Ac is then transferred to compound 21 (also shown in
- Figure 1 as compounds 4a,b) in the last step by the rat liver Gal( ⁇ 1-3/4) GlcNAc ⁇ 2-3 sialyltransferase 60 providing hexasaccharide 22 (also shown in Figure 1 as compounds 5a,b).
- compound 19 can be made in a totally synthetic scheme
- compound 19 can be obtained by total chemical
- R can be an aglycone or a saccharide moiety itself attached to an aglycone.
- a wide variety of glycosylation methods are available in order to synthesize ⁇ -glycosides.
- the present synthesis is derived from the synthesis described by Alais et al. 30 .
- glycosidases can be used, instead of glycosyltransferases, for the synthesis of glycosides in appropriate conditions.
- the main characteristics of the use of both types of enzymes have been reviewed by Ichikawa et al. 62 .
- sialidases 63,64 could be used to synthesize some of the saccharides.
- the blocked disaccharide glycosyl donor 11 30 is used in a glycosylation reaction of the desired alcohol ROH catalyzed by trimethylsilyltrifluoromethanesulfonate to lead to the glycoside 12.
- Mild de-O-acetylation provided 13.
- Disaccharide 13 is reacted with acetone in the presence of an acid catalyst, such as p-toluene sulfonic acid at 60oC, leading to a mixture of the 3,4- and of the 4,6-isopropylidene derivative 14 and 15 which are separated.
- the 3,4-isopropylidene derivative 14 is totally acetylated in pyridine with acetic anhydride and the isopropylidene group hydrolyzed in a mixture of acetic acid and water at 90°C providing 16.
- pentasaccharide obtained is identical to the same compound obtained earlier by using a different route 65 .
- the transformation is quantitative.
- Neu5Ac is transferred to pentasaccharide 21 by using the Gal( ⁇ 1-3/4)GlcNAc ⁇ (2-3)sialyltransferase from rat liver 25 .
- Another appropriate transferase from other sources can also be used. This step is performed according to the earlier report and provides the same product as described in Venot et al. 65 .
- Pentasaccharide 21 should be an acceptor for this transferase since the ⁇ -fucosyl residue is not linked to the penultimate GlcNAc moiety. Further sequential transfer of galactose and of Neu5Ac by the appropriate glycosyltransferase will lead to octasaccharides 26.
- these compounds affect the cell mediated immune response in a number of ways. Specifically, these compounds can inhibit the ability of the immune response to become educated about a specific antigen when the compound is administered simultaneously with the first exposure of the immune system to the antigen. Also,
- hexasaccharide glycosides I and IV can inhibit the effector phase of a cell-mediated immune response (eg., the inflammatory component of a DTH response) when administered after second or later exposures of the immune system to the same antigen.
- a cell-mediated immune response eg., the inflammatory component of a DTH response
- hexasaccharide glycosides I and IV can induce tolerance to antigens when administered at the time of second or later exposures of the immune system to the antigen.
- Hexasaccharide glycosides I and IV The suppression of the inflammatory component of the immune response by hexasaccharide glycosides I and IV is believed to require the initiation of a secondary immune response (i.e., a response to a second exposure to antigen).
- a secondary immune response i.e., a response to a second exposure to antigen.
- glycoside I or IV is generally administered to the patient at least about 0.5 hours after an
- inflammatory episode preferably, at least about 1 hour after, and most preferably, at least about 5 hours after an inflammatory episode or exacerbation.
- an antigen eg. the inflammatory component of a DTH response
- the specific dose employed is regulated by the particular cell-mediated immune response being treated as well as by the judgment of the attending clinician depending upon factors such as the severity of the adverse immune response, the age and general condition of the patient, and the like.
- glycosides I or IV is generally administered parenterally, such as intranasally,
- the suppression of a cell-mediated immune response eg. the inflammatory component of a DTH response
- the suppression of a cell-mediated immune response is reduced by at least about 10% as opposed to control measured 24 hours after administration of the challenge to the mammal and 19 hours after administration of hexasaccharide glycosides I or IV.
- hexasaccharide glycosides I or IV In addition to providing suppression of the inflammatory component of the cell-mediated immune response to an antigen, administration of the hexasaccharide glycosides I or IV also imparts a tolerance to additional challenges from the same antigen. In this regard, re-challenge by the same antigen weeks after administration of hexasaccharide glycosides I or IV results in a significantly reduced immune response.
- reducing sensitization means that the hexasaccharide glycosides I or IV, when administered to a mammal in an effective amount along with a sufficient amount of antigen to induce an immune response, reduces the ability of the immune system of the mammal to become educated and thus sensitized to the antigen administered at the same time as hexasaccharide glycosides I or IV.
- an "effective ⁇ imount" of this compound is that amount which will reduce sensitization (immunological education) of a mammal, including humans, to an antigen administered simultaneously as determined by a reduction in a cell-mediated response to the antigen such as DTH responses as tested by the footpad challenge test.
- the reduction in sensitization will be at least about 20% and more preferably at least about 30% or more.
- hexasaccharide glycosides I or IV are effective in reducing sensitization when administered at a dosage range of from about 0.5 mg to about 50 mg/kg of body weight, and preferably from about 0.5 mg to about 5 mg/kg of body weight.
- the specific dose employed is regulated by the sensitization being treated as well as the judgement of the attending clinician depending upon the age and general condition of the patient and the like.
- “Simultaneous" administration of the compound with the antigen with regard to inhibiting sensitization means that the compound is administered once or continuously throughout a period of time within 3 hours of the administration of an antigen, more preferably the compound is administered within 1 hour of the antigen.
- compositions suitable for use in the parenteral administration of an effective amount of hexasaccharide glycosides I or IV comprise a
- suitable pharmaceutical compositions can additionally contain optional components such as an adjuvant, a preservative, etc.
- suitable pharmaceutical compositions can include oral compositions, transdermal compositions or bandages etc., which are well known in the art.
- Hexasaccharide glycosides I and IV containing an analogue of sialic acid are also useful in preparing artificial antigens which can cross-react with antigenic determinants having a similar
- CMP-Neu5Ac cytidine-5'-monophospho-N- acetylneuraminic acid
- DTH delayed-type hypersensitivity
- Millipore Millipore Corp., Bedford, MA.
- latrobeads (6RS-8060) were from latron Laboratories, Tokyo, Japan and the AG 50W X 8 ion exchange resin was purchased from BioRad, Richmond, California.
- CMP-Neu5Ac was purchased from Sigma Chemical Company (St. Louis, Missouri) and GDP-fucose was obtained by chemical synthesis.
- 59 ⁇ Gal(1-4) ⁇ GlcNAc(1-3) ⁇ Gal- (1-4) ⁇ GlcNAc-OR was obtained by following the procedures of Alais et al 30 with the appropriate substitution of the aglycon. Evaporation of organic solvents was done at 20-25oC using a rotory
- the rat liver ⁇ Gal(1-3/4) ⁇ GlcNAc ⁇ (2-3)sialyltransferase (EC 2.4.99.5) was purified by affinity chromatography according to the procedure of Mazid, et al. 19 but using a matrix obtained by covalently linking the hapten ⁇ Gal(1-3) ⁇ GlcNAcO(CH 2 ) 8 CO 2 H 66 activated as in its N-succinimidyl ester to
- the enzymatic sialylations were carried out at 37' C in a plastic tube using a sodium cacodylate buffer (50 mM, pH 6.5) containing Triton CF-54
- the ⁇ GlcNAc ⁇ (1-3/4) fucosyltranferase was purified from human milk, as reported. 4 The
- Example 3 Preparation of 8-Methoxycarbonyloctyl ⁇ -D-galactopyranosyl-(1-4)-O-2-acetamido- 2-deoxy- ⁇ -D-glucopyranosyl-(1-3)-O- ⁇ -D- galactopyranosyl-(1-4)-O-[ ⁇ -L-fucopyranosyl-(1-3)-O-]2-acetamido-2-deoxy- ⁇ - D-glucopyranoside (4a) and the 8- carboxyoctyl glycoside (4b)
- the rat liver ⁇ Gal(1-3/4) ⁇ GlcNAc ⁇ (2-3.)sialyl ⁇ transferase (EC 2.4.99.5) was purified by affinity chromatography according to the procedure of Mazid, et al. 19 but using a matrix obtained by covalently linking the hapten ⁇ Gal(1-3)BGlcNAcO(CH 2 ) 8 CO 2 H 66 activated as in its N-succinimidyl ester to
- the enzymatic sialylations were carried out at 37°C in a plastic tube using a sodium cacodylate buffer (50 mM, pH 6.5) containing Triton CF-54 (0.5%), BSA (1 mg/mL) and calf intestine alkaline phosphatase. 69
- the final reaction mixtures were diluted with H 2 O and applied onto C 18 Sep-Pak cartridges as reported. 4 After washing with H 2 O, the products were eluted with CH 3 OH and the solvents evaporated. The residue was dissolved in a 65:35:5 mixture of CHCl 3 , CH 3 OH and H 2 O and applied on a small column of latrobeads (0.200 to 0.500 g).
- the ⁇ GlcNAc ⁇ (1-3/4)fucosyltransferase (EC 2.4.1.65) was purified from human milk, as reported. 4 The enzymatic reactions were carried out at 37oC in a plastic tube using a sodium cacodylate buffer (100 mM, pH 6.5), MnCl 2 (10 mM), ATP (1.6 mM), NaN 3 (1.6 mM). The reaction products were isolated and purified as indicated above.
- Tetra-n-butylammonium hydroxide (40% aq. w/w, about 150g) was added dropwise to a solution of phosphoric acid (85% aq, w/w, 18g, 0.155 mmol) in water (150 mL) until the pH reached 7. Water was then evaporated in vacuo to give a syrup which was co-evaporated with dry aceto-nitrile (2 ⁇ 400 mL) followed by dry toluene (2 ⁇ 400 mL). The resulting white solid (75g) was dried in vacuo and stored over phosphorus pentoxide under vacuum until used.
- Guanosine 5'-( ⁇ -1-fucopyranosyl)-diphosphate was prepared from ⁇ -L-fucopyranosyl-1-phosphate using two different art recognized procedures as set forth below:
- ⁇ -L-fucopyranosyl-1-phosphate and guanosine 5'-mono-phosphomorpholidate (4-morpholine-N,N'-dicyclohexyl-carboxamidine salt, available from Sigma, St. Louis, Missouri, "GMP-morpholidate") were reacted as described in a recent modification 74,75 of Nunez's original procedure 72 .
- tri-n-octylamine (0.800g, available from Aldrich Chemical Company, Milwaukee, Wisconsin) was added to a mixture of ⁇ -L-fucopyranosyl-1-phosphate (triethylammonium salt, 1.00g, about 2.20 mmol) in dry pyridine (10 mL) under nitrogen the solvent removed in vacuo. The process was repeated three times with care to allow only dry air to enter the flask.
- GMP morpholidate (2.4g, about 3.30 mmol) was dissolved in a 1:1 mixture of dry dimethylformamide and pyridine (10 mL). The solvents were evaporated in vacuo and the procedure repeated three times as above. The residue was dissolved in the same mixture of solvents (20 mL) and the solution added to the reaction flask accompanied by crushed
- Fractions 29 to 45 and 47 to 57 were separately pooled and freeze-dried providing recovered ⁇ -L-fuco-pyranosyl-1-phosphate (0.264g and 0.223g, respectively, in which the second fraction contains some impurities). Occasionally, pooling of
- guanosine 5'-( ⁇ -1-fucopyranosyl)-diphosphate in good purity ( 1 H-n.m.r.).
- all the material enriched in guanosine 5'-( ⁇ -1-fuco-pyranosyl)-diphosphate was dissolved in a minimum amount of water and run on the same column which had been regenerated by washing with large amounts of
- ⁇ -L-fucopyranosyl-1-phosphate and guanosine 5'-monophosphomorpholidate (4-morphollne-N,N'-dicyclohexyl-carboxamidine salt — "GMP-morpholidate") were reacted in dry pyridine as indicated in the original procedure 72 . Accordingly, the ⁇ -L-fucopyranosyl-1-phosphate (triethyl-ammonium salt, 0.528g, about 1.18 mmol) was dissolved in dry pyridine (20 mL) and the solvent removed in vacuo. The process was repeated three times with care to allow only dry air to enter the flask.
- GMP-morpholidate (1.2g, 1.65 mmol) and pyridine (20 mL) were added into the reaction flask, the solvent evaporated in vacuo and the process repeated three times as above. Pyridine (20 mL) was added to the final residue and the heterogeneous mixture was stirred for 3 to 4 days at room temperature under nitrogen. An insoluble mass was formed which had to be occasionally broken down by sonication.
- the bovine milk ⁇ GlcNAc ⁇ (1-4) galactosyltransferase (EC 2.4.1.22, specific activity 6.5 units/mg of protein) and UDP-Gal were obtained from Sigma.
- the enzymatic reactions were carried out at 37° in a plastic tube using a sodium cacodylate buffer (100 mM, pH 7.5) containing 20 mM manganese dichloride.
- the reaction products were purified as indicated above in the case of the preparative sialylation.
- the terminal methyl ester of the aglycone is hydrolyzed.
- the final products may possibly be isolated as saccharides possessing the aglycone terminated by a methyl ester of a free acid group. These two saccharides are separated during the step of the chromatography on latrobeads as indicated above.
- the two forms of the aglycone of the saccharide are identified by 1 H-n.m.r.
- dichloromethane (4 mL) was added to the mixture of the disaccharide donor 11 30 (2.0 g, 2.6 mmol), drierite (4.0 g, crushed) and 8-methoxycarbonyl- octanol (1.9 g, 10.0 mmol) in dichloromethane (30 mL) at 4°C. After stirring for 0.5 h at 4°C, the mixture was slowly warmed up to room temperature for 1 h. After cooling to 4°C, a second portion of the catalyst (0.250 mL, 1.3 mmol) in dichloromethane (2 mL) was added. After slowly warming up and stirring at room temperature for 1 h, the reaction was stopped by addition of triethylamine. After
- Trimethylsilyltrifluoromethanesulfonate (0.036 mL, 0.060 mmol) in methylene chloride (0.5 mL) was added to a solution of compound 16 (0.100 g, 0.123 mmol) in methylene chloride (5 mL).
- a solution of the imidate 17 (0.102 g, 0.185 mmol) in methylene chloride (4 mL) was slowly added to the above solution cooled at -70°. The mixture was further stirred at that temperature for 0.5 h.
- Example 8 Synthesis of 8-Methoxycarbonyloctyl (5- acetamido-3,5-dideoxy- ⁇ -D-glycero-D- galacto-2-nonulopyranosylonic acid)-(2-3)- O-( ⁇ -D-galactopyranosyl)-(1-4)-O-(2- acetamido-2-deoxy- ⁇ -D-glucopyranosyl)-(1- 3)-O-( ⁇ -D-galactopyranosyl)-(1-4)-O-[ ⁇ -L- fucopyranosyl-(1-3)-O]-2-acetamido-2- deoxy- ⁇ -D-glucopyranoside (22) (Compound 22 — the CD-65/VIM-2 Saccharide) A.
- Examples 9 and 10 illustrate the immunosuppressive properties of hexasaccharide glycoside 5a.
- Example 9 Inhibition of DTH Inflammatory Response
- DTH inflammatory responses were measured using the mouse footpad swelling assay as described by Smith and Ziola 31 . Briefly, groups of Balb/c mice were immunized with 10 ⁇ g of the L111 S-Layer protein, a bacterial surface protein 32 from Clostridium thermohvdrosulfuricum L111-69 which has been shown to induce a strong inflammatory DTH response. Seven days later, each group of mice was footpad-challenged with 10 ⁇ g of L-111 S-Layer protein. The resulting inflammatory footpad swelling was measured with a Mitutoyo Engineering micrometer 24 hours after challenge.
- mice received 100 ⁇ g of this compound, injected into the tail vein, 5 hours after challenge.
- Control groups received 100 ⁇ L of phosphate-buffered saline (PBS).
- PBS phosphate-buffered saline
- mice injected with hexasaccharide glycoside 5a had less than 50% of the footpad swelling as compared to the control mice.
- Example 10 Persistence of Suppression of the DTH Inflammatory Response at 11 Weeks After Challenge
- mice treated with hexasaccharide glycoside 5a in Example 7 were re- challenged with L111 S-Layer protein 11 weeks after primary immunization.
- Mice treated with the PBS control responded with the usual degree of footpad swelling whereas mice treated with hexasaccharide glycoside 5a showed a reduction in footpad swelling of about 40%, i.e., the mice treated with hexasaccharide glycoside 5a exhibited only about 60% of the footpad swelling exhibited in mice treated with PBS.
- This example examines whether hexasaccharide 5a could inhibit ELAM-1 dependent cell adhesion to activated vascular endothelium.
- an in vitro cell binding assay was preformed as described by Lowe et al 33 . Briefly, human umbilical vein endothelial cells (HUVECs purchased from Cell Systems, Seattle, WA, USA) were stimulated with TNF ⁇ (10 ng/ml) to express ELAM-1 .
- Human tumor cell lines, U937 or HL60 which have been shown to bind to HUVECs, in an ELAM-1 dependent manner were used to measure the effect that hexasaccharide glycoside 5a has on the ELAM-1 dependent binding to the HUVEC.
- the results of this example demonstrate that hexasaccharide glycoside 5a inhibits ELAM-1 dependent binding to the HUVECs.
- Example 9 establish the effectiveness of the hexasaccharide glycosides described herein in treating immune responses to an antigen and in inducing tolerance to the antigen in a mammal (mice).
- a mammal such hexasaccharide glycosides will also be effective in treating human immune responses.
- Example 11 establishes that hexasaccharide glycoside 5a inhibits ELAM-1 dependent binding to the HUVEC.
- hexasaccharide glycosides of formula I and IV above could be used to suppress a cell-mediated immune response to an antigen by mere substitution for hexasaccharide glycoside 5a described in these examples.
- the compounds defined by formula II, III, V and VI are useful at least as intermediates in the preparation of compounds I and IV.
- the transfer of L-fucose via a (1-3) fucosyltransferase can employ a compatible analog of L-fucose which is recognized by the transferase and provides for products wherein Y and Y' are compatible analogs of L-fucose.
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Abstract
On décrit des méthodes de préparation de dérivés à monofucosylation et sialylation du composé betaGal(1-4)betaGlcNAc(1-3)betaGal(1-4)betaGlcNAc-R. En particulier, les méthodes de ladite invention permettent une synthèse en plusieurs étapes, la monofucosylation sélective étant réalisée sur le groupe 3-hydroxy sur une seule des unités GlcNAc trouvées dans le composé betaGal(1-4)betaGlcNAc(1-3)betaGal(1-4)betaGlcNAc-R. Dans cette étape, la monofucosylation est réalisée par l'utilisation de la alpha(1-3) fucosyltransférase.Methods for the preparation of monofucosylation and sialylation derivatives of the compound betaGal (1-4) betaGlcNAc (1-3) betaGal (1-4) betaGlcNAc-R are described. In particular, the methods of said invention allow a synthesis in several stages, the selective monofucosylation being carried out on the 3-hydroxy group on only one of the GlcNAc units found in the compound betaGal (1-4) betaGlcNAc (1-3) betaGal ( 1-4) betaGlcNAc-R. In this step, monofucosylation is achieved by the use of alpha (1-3) fucosyltransferase.
Description
Claims
Applications Claiming Priority (7)
Application Number | Priority Date | Filing Date | Title |
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US71416191A | 1991-06-10 | 1991-06-10 | |
US714161 | 1991-06-10 | ||
US77125991A | 1991-10-02 | 1991-10-02 | |
US771259 | 1991-10-02 | ||
US88901792A | 1992-05-26 | 1992-05-26 | |
US889017 | 1992-05-26 | ||
PCT/CA1992/000251 WO1992022662A1 (en) | 1991-06-10 | 1992-06-10 | Methods for the synthesis of monofucosylated oligosaccharides terminating in di-n-acetyllactosaminyl structures |
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EP0591256A1 true EP0591256A1 (en) | 1994-04-13 |
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EP92911470A Withdrawn EP0588852A1 (en) | 1991-06-10 | 1992-06-09 | Immunosuppressive and tolerogenic oligosaccharide derivatives |
EP92911443A Ceased EP0591256A1 (en) | 1991-06-10 | 1992-06-10 | Methods for the synthesis of monofucosylated oligosaccharides terminating in di-n-acetyllactosaminyl structures |
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EP (2) | EP0588852A1 (en) |
JP (2) | JPH07502011A (en) |
CA (2) | CA2110495A1 (en) |
WO (2) | WO1992022301A1 (en) |
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US5559103A (en) * | 1993-07-21 | 1996-09-24 | Cytel Corporation | Bivalent sialyl X saccharides |
US5631283A (en) * | 1994-02-02 | 1997-05-20 | The United States Of America As Represented By The Secretary Of The Army | Use of sialic acid or antibodies to sialidase as anti-infectious agents and anti-inflammatory agents |
DE69530910D1 (en) * | 1994-07-15 | 2003-07-03 | Taiyo Kagaku Co | MEDICAL COMPOSITIONS CONTAINING A SIALINIC DERIVATIVE |
DE69739750D1 (en) * | 1996-07-23 | 2010-03-18 | Seikagaku Kogyo Co Ltd | NEW LACTOSAMINE OLIGOSACCHARIDES AND METHOD FOR THE PRODUCTION THEREOF |
AU2002319207B2 (en) * | 2001-06-19 | 2007-03-01 | Reinhard Brossmer | Sialic acid derivatives for use as siglec inhibitors |
DE10204000A1 (en) * | 2002-02-01 | 2003-08-14 | Nutricia Nv | Sialysed carbohydrates |
EP1911850A1 (en) * | 2006-10-09 | 2008-04-16 | Centre National De La Recherche Scientifique (Cnrs) | Method of producing 6-thio-sialylated glycoproteins and glycoconjugates |
CN102046781A (en) | 2008-05-30 | 2011-05-04 | 动量制药公司 | Saccharide structures and methods of making and using such structures |
KR20160132034A (en) | 2014-03-13 | 2016-11-16 | 우니페르시테트 바젤 | Carbohydrate ligands that bind to igm antibodies aganist myelin-associated glycoprotein |
US11091591B2 (en) | 2015-09-16 | 2021-08-17 | Universität Basel | Carbohydrate ligands that bind to antibodies against glycoepitopes of glycosphingolipids |
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EP0380084A3 (en) * | 1989-01-27 | 1991-03-13 | The Biomembrane Institute | Method for the inhibition of cell-cell and cell substratum interactions by the blocking of carbohydrate-carbohydrate interactions |
EP0395217A3 (en) * | 1989-04-28 | 1991-01-23 | The Biomembrane Institute | Bio-organic synthesis of dimeric lex (difucosyl y2; iii3fucv3-funcnlc6cer) and analogues thereof |
US5109116A (en) * | 1989-07-24 | 1992-04-28 | Monsanto Company | Immunosuppressive glycoprotein |
CA2067250C (en) * | 1989-10-20 | 2008-06-17 | Bruce Furie | Inhibition of padgem-mediated cell binding |
AU8007791A (en) * | 1990-06-15 | 1992-01-07 | Cytel Corporation | Intercellular adhesion mediators |
US5211937A (en) * | 1990-07-30 | 1993-05-18 | Glycomed Incorporated | Method of determining a site of inflammation utilizing elam-1 ligands |
NZ240316A (en) * | 1990-10-25 | 1996-12-20 | Univ Michigan | Compound for treating disease mediated by the elaboration of elam-1 on endothelial cells |
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- 1992-06-09 CA CA002110495A patent/CA2110495A1/en not_active Abandoned
- 1992-06-09 JP JP4511195A patent/JPH07502011A/en active Pending
- 1992-06-09 WO PCT/CA1992/000244 patent/WO1992022301A1/en not_active Application Discontinuation
- 1992-06-09 EP EP92911470A patent/EP0588852A1/en not_active Withdrawn
- 1992-06-10 EP EP92911443A patent/EP0591256A1/en not_active Ceased
- 1992-06-10 JP JP4511199A patent/JPH06510661A/en active Pending
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JPH07502011A (en) | 1995-03-02 |
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