CN117285577A - Method for synthesizing acinetobacter baumannii surface sugar antigen with high stereoselectivity - Google Patents
Method for synthesizing acinetobacter baumannii surface sugar antigen with high stereoselectivity Download PDFInfo
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- CN117285577A CN117285577A CN202310514694.XA CN202310514694A CN117285577A CN 117285577 A CN117285577 A CN 117285577A CN 202310514694 A CN202310514694 A CN 202310514694A CN 117285577 A CN117285577 A CN 117285577A
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- 238000000034 method Methods 0.000 title claims abstract description 37
- 230000002194 synthesizing effect Effects 0.000 title claims abstract description 13
- 241000588626 Acinetobacter baumannii Species 0.000 title abstract description 18
- 239000000427 antigen Substances 0.000 title abstract description 12
- 102000036639 antigens Human genes 0.000 title description 5
- 108091007433 antigens Proteins 0.000 title description 5
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 60
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 56
- 125000005524 levulinyl group Chemical group 0.000 claims abstract description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 162
- 150000001875 compounds Chemical class 0.000 claims description 158
- 238000006243 chemical reaction Methods 0.000 claims description 57
- 238000004440 column chromatography Methods 0.000 claims description 57
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 52
- 239000000203 mixture Substances 0.000 claims description 37
- 238000001914 filtration Methods 0.000 claims description 36
- 238000010791 quenching Methods 0.000 claims description 20
- 239000003054 catalyst Substances 0.000 claims description 17
- 150000002016 disaccharides Chemical class 0.000 claims description 17
- 230000000171 quenching effect Effects 0.000 claims description 17
- 125000000025 triisopropylsilyl group Chemical group C(C)(C)[Si](C(C)C)(C(C)C)* 0.000 claims description 16
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 claims description 14
- 239000000047 product Substances 0.000 claims description 14
- 150000004043 trisaccharides Chemical class 0.000 claims description 12
- 239000003960 organic solvent Substances 0.000 claims description 10
- 239000000499 gel Substances 0.000 claims description 9
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 7
- 238000000967 suction filtration Methods 0.000 claims description 7
- 238000001308 synthesis method Methods 0.000 claims description 7
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 claims description 7
- UYWQUFXKFGHYNT-UHFFFAOYSA-N phenylmethyl ester of formic acid Natural products O=COCC1=CC=CC=C1 UYWQUFXKFGHYNT-UHFFFAOYSA-N 0.000 claims description 6
- 238000010898 silica gel chromatography Methods 0.000 claims description 6
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 4
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 claims description 3
- 125000003236 benzoyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)=O 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 claims description 3
- 238000010549 co-Evaporation Methods 0.000 claims description 2
- 230000006837 decompression Effects 0.000 claims description 2
- 239000012467 final product Substances 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims description 2
- 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 claims description 2
- LMBFAGIMSUYTBN-MPZNNTNKSA-N teixobactin Chemical compound C([C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H](CCC(N)=O)C(=O)N[C@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H]1C(N[C@@H](C)C(=O)N[C@@H](C[C@@H]2NC(=N)NC2)C(=O)N[C@H](C(=O)O[C@H]1C)[C@@H](C)CC)=O)NC)C1=CC=CC=C1 LMBFAGIMSUYTBN-MPZNNTNKSA-N 0.000 claims 1
- 125000006239 protecting group Chemical group 0.000 abstract description 19
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- 238000004458 analytical method Methods 0.000 abstract description 7
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- 239000001257 hydrogen Substances 0.000 abstract description 5
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- 239000003208 petroleum Substances 0.000 description 51
- 239000000243 solution Substances 0.000 description 47
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 46
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 42
- 239000000386 donor Substances 0.000 description 41
- 230000002829 reductive effect Effects 0.000 description 39
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 37
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 35
- 239000000370 acceptor Substances 0.000 description 33
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- 239000007787 solid Substances 0.000 description 31
- 238000003756 stirring Methods 0.000 description 31
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 30
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- PCLIMKBDDGJMGD-UHFFFAOYSA-N N-bromosuccinimide Chemical compound BrN1C(=O)CCC1=O PCLIMKBDDGJMGD-UHFFFAOYSA-N 0.000 description 14
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 14
- 239000000543 intermediate Substances 0.000 description 14
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 13
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- 239000007858 starting material Substances 0.000 description 13
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 13
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 12
- 239000005457 ice water Substances 0.000 description 12
- -1 methoxybenzyl Chemical group 0.000 description 12
- 238000000926 separation method Methods 0.000 description 12
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 11
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- 239000006260 foam Substances 0.000 description 10
- 230000000707 stereoselective effect Effects 0.000 description 10
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 9
- WLLIXJBWWFGEHT-UHFFFAOYSA-N [tert-butyl(dimethyl)silyl] trifluoromethanesulfonate Chemical compound CC(C)(C)[Si](C)(C)OS(=O)(=O)C(F)(F)F WLLIXJBWWFGEHT-UHFFFAOYSA-N 0.000 description 9
- 238000000746 purification Methods 0.000 description 9
- 239000012046 mixed solvent Substances 0.000 description 8
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 8
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 description 7
- 238000005580 one pot reaction Methods 0.000 description 7
- KDLHZDBZIXYQEI-UHFFFAOYSA-N palladium Substances [Pd] KDLHZDBZIXYQEI-UHFFFAOYSA-N 0.000 description 7
- 239000002994 raw material Substances 0.000 description 7
- FPGGTKZVZWFYPV-UHFFFAOYSA-M tetrabutylammonium fluoride Chemical compound [F-].CCCC[N+](CCCC)(CCCC)CCCC FPGGTKZVZWFYPV-UHFFFAOYSA-M 0.000 description 7
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 6
- 229960000583 acetic acid Drugs 0.000 description 6
- CSCPPACGZOOCGX-WFGJKAKNSA-N acetone d6 Chemical compound [2H]C([2H])([2H])C(=O)C([2H])([2H])[2H] CSCPPACGZOOCGX-WFGJKAKNSA-N 0.000 description 6
- 238000005273 aeration Methods 0.000 description 6
- 238000010276 construction Methods 0.000 description 6
- 229920001542 oligosaccharide Polymers 0.000 description 6
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 description 5
- 150000001720 carbohydrates Chemical group 0.000 description 5
- 238000007865 diluting Methods 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 150000004676 glycans Chemical class 0.000 description 5
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- 239000005017 polysaccharide Substances 0.000 description 5
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical class [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 5
- YRIZYWQGELRKNT-UHFFFAOYSA-N 1,3,5-trichloro-1,3,5-triazinane-2,4,6-trione Chemical compound ClN1C(=O)N(Cl)C(=O)N(Cl)C1=O YRIZYWQGELRKNT-UHFFFAOYSA-N 0.000 description 4
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 4
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 4
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 4
- LQZMLBORDGWNPD-UHFFFAOYSA-N N-iodosuccinimide Substances IN1C(=O)CCC1=O LQZMLBORDGWNPD-UHFFFAOYSA-N 0.000 description 4
- 229930182475 S-glycoside Natural products 0.000 description 4
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 description 4
- XXROGKLTLUQVRX-UHFFFAOYSA-N allyl alcohol Chemical compound OCC=C XXROGKLTLUQVRX-UHFFFAOYSA-N 0.000 description 4
- KQIADDMXRMTWHZ-UHFFFAOYSA-N chloro-tri(propan-2-yl)silane Chemical compound CC(C)[Si](Cl)(C(C)C)C(C)C KQIADDMXRMTWHZ-UHFFFAOYSA-N 0.000 description 4
- 239000000463 material Substances 0.000 description 4
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 4
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- BCNZYOJHNLTNEZ-UHFFFAOYSA-N tert-butyldimethylsilyl chloride Chemical compound CC(C)(C)[Si](C)(C)Cl BCNZYOJHNLTNEZ-UHFFFAOYSA-N 0.000 description 4
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- HVHZEKKZMFRULH-UHFFFAOYSA-N 2,6-ditert-butyl-4-methylpyridine Chemical compound CC1=CC(C(C)(C)C)=NC(C(C)(C)C)=C1 HVHZEKKZMFRULH-UHFFFAOYSA-N 0.000 description 3
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- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 3
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 3
- 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 3
- JRMUNVKIHCOMHV-UHFFFAOYSA-M tetrabutylammonium bromide Chemical compound [Br-].CCCC[N+](CCCC)(CCCC)CCCC JRMUNVKIHCOMHV-UHFFFAOYSA-M 0.000 description 3
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- NWZSZGALRFJKBT-KNIFDHDWSA-N (2s)-2,6-diaminohexanoic acid;(2s)-2-hydroxybutanedioic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O.NCCCC[C@H](N)C(O)=O NWZSZGALRFJKBT-KNIFDHDWSA-N 0.000 description 2
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- MSWZFWKMSRAUBD-UHFFFAOYSA-N 2-Amino-2-Deoxy-Hexose Chemical compound NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 2
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 2
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- 241000568569 Acinetobacter baumannii ATCC 17978 Species 0.000 description 2
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- JGFBRKRYDCGYKD-UHFFFAOYSA-N dibutyl(oxo)tin Chemical compound CCCC[Sn](=O)CCCC JGFBRKRYDCGYKD-UHFFFAOYSA-N 0.000 description 2
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- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- DUKURNFHYQXCJG-UHFFFAOYSA-N Lewis A pentasaccharide Natural products OC1C(O)C(O)C(C)OC1OC1C(OC2C(C(O)C(O)C(CO)O2)O)C(NC(C)=O)C(OC2C(C(OC3C(OC(O)C(O)C3O)CO)OC(CO)C2O)O)OC1CO DUKURNFHYQXCJG-UHFFFAOYSA-N 0.000 description 1
- 201000009906 Meningitis Diseases 0.000 description 1
- 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 description 1
- 101150003085 Pdcl gene Proteins 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 208000000474 Poliomyelitis Diseases 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- KEAYESYHFKHZAL-UHFFFAOYSA-N Sodium Chemical compound [Na] KEAYESYHFKHZAL-UHFFFAOYSA-N 0.000 description 1
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 1
- 206010062255 Soft tissue infection Diseases 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- LPTITAGPBXDDGR-IWQYDBTJSA-N [(2r,3r,4s,5r)-3,4,5,6-tetraacetyloxyoxan-2-yl]methyl acetate Chemical compound CC(=O)OC[C@H]1OC(OC(C)=O)[C@H](OC(C)=O)[C@@H](OC(C)=O)[C@@H]1OC(C)=O LPTITAGPBXDDGR-IWQYDBTJSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- IAJILQKETJEXLJ-QTBDOELSSA-N aldehydo-D-glucuronic acid Chemical compound O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C(O)=O IAJILQKETJEXLJ-QTBDOELSSA-N 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 229940124350 antibacterial drug Drugs 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- PASDCCFISLVPSO-UHFFFAOYSA-N benzoyl chloride Chemical compound ClC(=O)C1=CC=CC=C1 PASDCCFISLVPSO-UHFFFAOYSA-N 0.000 description 1
- 125000000649 benzylidene group Chemical group [H]C(=[*])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- UWTDFICHZKXYAC-UHFFFAOYSA-N boron;oxolane Chemical compound [B].C1CCOC1 UWTDFICHZKXYAC-UHFFFAOYSA-N 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- SBTSVTLGWRLWOD-UHFFFAOYSA-L copper(ii) triflate Chemical compound [Cu+2].[O-]S(=O)(=O)C(F)(F)F.[O-]S(=O)(=O)C(F)(F)F SBTSVTLGWRLWOD-UHFFFAOYSA-L 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 125000000600 disaccharide group Chemical group 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229940097043 glucuronic acid Drugs 0.000 description 1
- 238000005858 glycosidation reaction Methods 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 239000000937 glycosyl acceptor Substances 0.000 description 1
- 125000003147 glycosyl group Chemical group 0.000 description 1
- JEGUKCSWCFPDGT-UHFFFAOYSA-N h2o hydrate Chemical compound O.O JEGUKCSWCFPDGT-UHFFFAOYSA-N 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 229920000669 heparin Polymers 0.000 description 1
- 229960002897 heparin Drugs 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 125000002883 imidazolyl group Chemical group 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000002493 microarray Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229950006780 n-acetylglucosamine Drugs 0.000 description 1
- 125000004923 naphthylmethyl group Chemical group C1(=CC=CC2=CC=CC=C12)C* 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- YWWARDMVSMPOLR-UHFFFAOYSA-M oxolane;tetrabutylazanium;fluoride Chemical compound [F-].C1CCOC1.CCCC[N+](CCCC)(CCCC)CCCC YWWARDMVSMPOLR-UHFFFAOYSA-M 0.000 description 1
- 238000005554 pickling Methods 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 235000015497 potassium bicarbonate Nutrition 0.000 description 1
- 229910000028 potassium bicarbonate Inorganic materials 0.000 description 1
- 239000011736 potassium bicarbonate Substances 0.000 description 1
- TYJJADVDDVDEDZ-UHFFFAOYSA-M potassium hydrogencarbonate Chemical compound [K+].OC([O-])=O TYJJADVDDVDEDZ-UHFFFAOYSA-M 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000007142 ring opening reaction Methods 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 208000013223 septicemia Diseases 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000012312 sodium hydride Substances 0.000 description 1
- 229910000104 sodium hydride Inorganic materials 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 229950009390 symclosene Drugs 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- ILMRJRBKQSSXGY-UHFFFAOYSA-N tert-butyl(dimethyl)silicon Chemical group C[Si](C)C(C)(C)C ILMRJRBKQSSXGY-UHFFFAOYSA-N 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- PVFOMCVHYWHZJE-UHFFFAOYSA-N trichloroacetyl chloride Chemical compound ClC(=O)C(Cl)(Cl)Cl PVFOMCVHYWHZJE-UHFFFAOYSA-N 0.000 description 1
- SEDZOYHHAIAQIW-UHFFFAOYSA-N trimethylsilyl azide Chemical compound C[Si](C)(C)N=[N+]=[N-] SEDZOYHHAIAQIW-UHFFFAOYSA-N 0.000 description 1
- 125000000026 trimethylsilyl group Chemical group [H]C([H])([H])[Si]([*])(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 208000019206 urinary tract infection Diseases 0.000 description 1
- 150000008135 α-glycosides Chemical class 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
-
- 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
-
- 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
- C07H15/18—Acyclic radicals, substituted by carbocyclic rings
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H23/00—Compounds containing boron, silicon, or a metal, e.g. chelates, vitamin B12
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Polymers & Plastics (AREA)
- Crystallography & Structural Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Materials Engineering (AREA)
- Saccharide Compounds (AREA)
Abstract
The invention belongs to the technical field of organic synthesis, and particularly relates to a method for synthesizing a surface glycoantigen of Acinetobacter baumannii with high stereoselectivity. The invention adopts proper protecting group and proper synthesis strategy, firstly utilizes inverse synthesis analysis method to determine [2+1+2] synthesis strategy of key pentasaccharide 2-70 and 2-69, then utilizes levulinyl (Lev) remote hydrogen bond induction method of adjacent saccharide to construct beta glycosidic bond, solves the problem of stereoselectivity of receptor with high steric hindrance and low reactivity, realizes optimization of synthesis condition of pentasaccharide 2-70 and 2-69 and improvement of stereoselectivity, and finally realizes synthesis of pentasaccharide by removing levulinyl protection. The invention completes the synthesis of the Acinetobacter baumannii ATCC 17961 lipopolysaccharide O-antigen, is convenient for knowing the influence of the sugar sequence and the length on antigenicity, and lays a foundation for the development of the Acinetobacter baumannii sugar vaccine.
Description
Technical Field
The invention belongs to the technical field of organic synthesis, and particularly relates to a method for synthesizing a surface glycoantigen of Acinetobacter baumannii with high stereoselectivity.
Background
Most of the polysaccharides used in current clinically available bacterial polysaccharide and glycoconjugate vaccines are extracted and purified from bacterial cell cultures, which are heterogeneous and easily vary from batch to batch. In contrast, the glycoconjugate vaccine obtained by the chemical synthesis method has high consistency and better safety. Furthermore, the use of chemical synthesis methods enables selection of codes representing repeat units, changes in length, and improvements in the design of synthetic oligosaccharide targets, all of which help to develop more stable glycoconjugate vaccines.
Key to oligosaccharide synthesis is the stereoselective construction of glycosidic linkages. Common oligosaccharide synthesis methods include linear synthesis, convergent synthesis and one-pot synthetic strategies emerging in recent years. One pot synthesis strategy allows for multiple steps of glycosylation reactions to be carried out continuously in one pot by modulating the activity of the glycosyl donor or acceptor. Compared with the traditional linear and convergent synthesis, the one-pot synthesis can avoid the adjustment of protecting groups between glycosylation reactions and the separation and purification of intermediates, and the synthesis efficiency is obviously improved.
Stereochemical control is critical during glycosylation reactions. Ortho-group participation is one of the most common strategies for directing the stereochemistry of newly formed glycosidic bonds. A typical glycosylation strategy is to mount a participating group on the C-2 atom near the center of the anomeric position of the glycosyl donor, with adjacent groups helping to stabilize the positive charge being formed at the anomeric position and to stereoscopically direct the formation of the 1, 2-trans glycosidic bond upon donor activation.
For a remote participation assistance strategy, a suitably designed remote directed acyl group from the C-3, C-4 or C-6 position, respectively, in a donor can achieve high 1, 2-cis selectivity during glycosylation. The remote participation of levulinyl (Lev) groups in ancillary glycosidic linkages can be used in highly stereoselective synthetic methods. All examples of remote group involvement currently relate to groups on the same sugar ring of the donor that are activated during glycosylation, while stereotactic involvement is not limited to groups within the activated sugar ring.
Acinetobacter baumannii (Acinetobacter baumannii) is an aerobic gram-negative bacillus, is listed by the world health organization as a key target spot for urgent need of novel antibacterial drugs, and is a common cause of soft tissue and urinary tract infection, septicemia, pneumonia and meningitis. With the increasing number of multi-drug resistant strains and their worldwide popularity, vaccines are an effective means of preventing and controlling acinetobacter baumannii infection. In the process of exploring and solving the problem of acinetobacter baumanii infection, capsular polysaccharide, lipopolysaccharide and lipooligosaccharide or O-glycan become potential immune targets for vaccine development.
Because of the specificity of the structure and important potential immunocompetence, the invention designs and synthesizes Acinetobacter baumannii ATCC 17961 lipopolysaccharide O-antigen (comprising tetraose with different branches, pentasaccharide containing single repeating unit and decasaccharide containing two repeating units). The oligosaccharide molecules can be used for screening immunogenic epitopes, and can also be used for comparing with the acinetobacter baumannii ATCC 17978 capsular polysaccharide O-antigen to know the influence of the sugar sequence and length on antigenicity, thereby laying a foundation for screening related sugar antigens of the acinetobacter baumannii.
Disclosure of Invention
The invention aims to provide a method for synthesizing a surface sugar antigen of Acinetobacter baumannii A.baumannii.ATCC 17961 with high stereoselectivity, which can efficiently synthesize pentasaccharide and decasaccharide and provides a raw material basis for developing a glycoconjugate vaccine.
In order to achieve the technical purpose, the invention adopts the following technical scheme:
the invention firstly provides a tetraose compound, which comprises a compound shown in a formula I and a compound shown in a formula II, and the specific structure is as follows:
further, R 1 TIPS, bn or H.
Further, ac is acetyl, bn is benzyl, TIPS is triisopropylsilyl, bz is benzoyl, TCA is trichloroacetic acid, and Cbz is benzyloxycarbonyl.
Specifically, the tetraose compound shown in the formula II is 2-13, 2-42 or 2-43, and the structural formula is as follows:
the invention further provides a pentasaccharide compound which comprises a compound shown in a formula III, and the specific structure is as follows:
further, R 2 Is Lev, bn or H; r is R 3 Is Lev, bn or H; r is R 4 Is TBS (Tunnel boring system),Or->
Further, bn is benzyl, lev is levulinyl, ph is phenyl, TIPS is triisopropylsilyl, TBS is tert-butyldimethylsilyl, TCA is trichloroacetic acid, cbz is benzyloxycarbonyl.
Specifically, the compound shown in the formula III is 2-70, 2-22, 2-69 or 2-23, and the structural formula is as follows:
furthermore, based on a general inventive concept, the invention also provides a synthesis method of the pentasaccharide compound shown in formula III, wherein the method realizes construction of glycosidic bond by levulinyl (Lev) of adjacent saccharide through remote hydrogen bond induction, and simultaneously improves stereoselectivity, and specifically comprises the following steps:
1) Mixing disaccharide donor, trisaccharide acceptor and toluene for co-evaporation to obtain a mixture, adding an organic solvent I into the mixture for pre-reaction, adding a catalyst I into the mixture for reaction at room temperature (20-25 ℃) until TLC analysis shows that the acceptor is completely consumed;
2) After the reaction, quenching, suction filtration, decompression concentration to obtain coarse product, and purifying with gel column chromatography and silica gel column chromatography successively to obtain final product.
Specifically, the disaccharide donor is 2-21, and the structural formula is:
specifically, the trisaccharide receptor is 2-72 and 2-49, and the structural formula is as follows:
specifically, the pre-reaction process in step 1) is to add the molecular sieve after the mixture is mixed with the organic solvent I, stir the mixture at room temperature for 10 minutes under an inert atmosphere, and stir the mixture at 0 ℃ for 5 minutes.
Specifically, the reaction time in step 1) is 2 to 6 hours, preferably 4 hours.
Specifically, the inert atmosphere is formed by adopting nitrogen or argon.
Specifically, the organic solvent I is dichloromethane, toluene or chloroform, preferably Dichloromethane (DCM).
Specifically, the catalyst I is TMSOTF (trimethyl silicone triflate).
Specifically, the molar ratio of disaccharide donor to trisaccharide acceptor is (1-2): 1, a step of; preferably 2:1.
Specifically, the molar ratio of the trisaccharide acceptor to the catalyst I is (1.6-1.9): 1, a step of; preferably 1.838:1.
Specifically, the volume ratio of the organic solvent I to the catalyst I is (1-1.5): 1, a step of; preferably 1.2:1.
Further, based on a general inventive concept, the present invention also provides an application of the pentasaccharide compound in synthesizing a decasaccharide compound.
Specifically, the decasaccharide compound comprises a compound shown in a formula IV and a compound shown in a formula V, and has the following structure:
Further, R 2 Is Lev, bn or H; r is R 3 Is Lev, bn or H.
Further, bn is benzyl, lev is levulinyl, TIPS is triisopropylsilyl, TCA is trichloroacetic acid, and Cbz is benzyloxycarbonyl.
Specifically, the compounds shown in the formula V are 2-24 and 2-75, and the structural formula is as follows:
further, the present invention also provides a method for synthesizing a decasaccharide compound using the pentasaccharide compound of formula III, the synthesis method comprising:
reacting pentasaccharide acceptor and pentasaccharide donor at 20-25deg.C for 2-4 hr under the condition of catalyst, quenching, filtering, concentrating, separating and purifying to obtain decasaccharide compound.
Specifically, the synthesis method of the decasaccharide compound 2-24 comprises the following steps:
mixing the pentasaccharide acceptor 2-22 and the pentasaccharide donor 2-23 with an organic solvent II and a catalyst II under inert atmosphere, carrying out oil bath reaction for 2-4h (preferably 3.5 h) at 20-25 ℃ (preferably 20 ℃), quenching, filtering, concentrating to obtain a crude product, and purifying by gel column chromatography and silica gel column chromatography in sequence to obtain the decasaccharide compound 2-24.
Specifically, the inert atmosphere is formed by adopting nitrogen or argon.
Specifically, the organic solvent II is DCM and Et 2 Mixtures of O wherein DCM and Et 2 The volume ratio of O is 1:3.
Specifically, the catalyst II is TBSOTf (tert-butyldisilyl triflate).
Specifically, the molar ratio of the pentasaccharide acceptor 2-22 to the pentasaccharide donor 2-23 is 1 (1-1.5); preferably 1:1.3.
Specifically, the molar ratio of pentasaccharide acceptor 2-22 to catalyst II is (2-5): 1, a step of; preferably 5:1.
Specifically, the volume ratio of the organic solvent II to the catalyst II is preferably 1000:1.
Further, the present invention also provides a method for synthesizing compound 2-75 using compound 2-24, comprising the steps of:
dissolving compound 2-24 in dichloromethane, stirring, sequentially adding pyridine and acetic acid, and adding N under ice bath 2 H 4 ·H 2 And (3) reacting the mixture of O and AcOH for 6 hours at room temperature, diluting, pickling, washing with a salt solution, drying, concentrating, and separating and purifying by column chromatography to obtain a foamy solid 2-75. Wherein, the compounds 2-24 and N 2 H 4 ·H 2 The mixture ratio of the O and the AcOH is 1000g to 1L. N (N) 2 H 4 ·H 2 The volume ratio of O to AcOH is 1:1.
Further, the present invention also provides a method for synthesizing compound 2-5 using compound 2-75, comprising the steps of:
dissolving compound 2-75 in tetrahydrofuran, adding tert-butanol and H 2 O、AcOH、Pd(OH) 2 C, under ice bath H 2 Ventilating for 15min, and then at H of 1atm 2 Reacting at room temperature for 5 days under the environment, filtering, concentrating under reduced pressure, and separating by using a Sephadex LH-20 column to obtain the compound 2-5. Wherein Pd (OH) 2 The mass ratio of the component (C) to the compound (2-75) is 5:1.
Compared with the prior art, the invention has the advantages that:
the invention completes the synthesis of the Acinetobacter baumannii ATCC 17961 lipopolysaccharide O-antigen, including tetraose, pentaose and decaose molecules by selecting proper protecting groups and proper synthesis strategies. The invention firstly utilizes the inverse synthetic analysis method to determine the [2+1+2] synthesis strategy of the key pentasaccharide 2-70 and 2-69, then utilizes the method of constructing beta glycosidic bond by the levulinyl (Lev) remote hydrogen bond induction of adjacent saccharides, solves the problem of high steric hindrance and stereoselectivity of low-reactivity receptors, realizes the optimization of the synthesis conditions of the pentasaccharide 2-70 and 2-69 and the improvement of the stereoselectivity, and finally realizes the synthesis of the pentasaccharide by removing the levulinyl protection.
The invention completes the synthesis of the Acinetobacter baumannii ATCC 17961 lipopolysaccharide O-antigen, is convenient to compare with the Acinetobacter baumannii ATCC 17978 capsular polysaccharide O-antigen, knows the influence of the sugar sequence and length on antigenicity, and lays a foundation for the development of the Acinetobacter baumannii sugar vaccine.
Detailed Description
The technical scheme of the present invention is described in further detail below by way of specific examples, but the scope of the present invention is not limited thereto.
Short for the following examples: ac is acetyl, ac 2 O is acetic anhydride, acOH is glacial acetic acid, all is allyl, bn is benzyl, bu 2 SnO is di-n-butyltin oxide, bz is benzoyl, cbz is carbobenzoxy, DCM is dichloromethane, DDQ is 2, 3-dichloro-5, 6-dicyano-1, 4-benzoquinone, DIPEA is diisopropylethylamine, DMAP is 4, 4-dimethylaminopyridine, DMF is dimethylformamide, DTBMP is 2, 6-di-tert-butyl-4-methylpyridine, EDCI is 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide, et is ethyl, im is imidazole, i- pr is isopropyl, me is methyl, MS is molecular sieve, nap is 2- (bromomethyl) naphthalene, NBS is N-bromosuccinimide, NIS is N-iodosuccinimide, PE is petroleum ether, EA is ethyl acetate, ph is phenyl, piVOH is trimethylacetic acid, PMB is methoxybenzyl, PMBCl is 4-methoxybenzyl chloride, PTFAI is N-phenyltrifluoroacetyl, PTFAICl is N-phenyltrifluoroacetyl chloride, PPY is 4-pyrrolidinylpyridine, py is pyridine, t bu is tert-butyl, TBAB is tetrabutylammonium bromide, TBAI is tetrabutylammonium iodide, TBAN is tetrabutylammonium nitrate, TBAF is tetrabutylammonium fluoride, TBDPS is tert-butyldiphenylsilyl, TBDPSCl is diphenyl tert-butyl Chlorosilane, TEMPO is 2, 6-tetramethylpiperidine oxide, BAIB is bis (acetoxy) iodobenzene, TBS is t-butyldimethylsilyl, TBSCl is dimethyl t-butylchlorosilane, TCA is trichloroacetic acid, tf is triflate, TIPS is triisopropylsilyl, tipcl is triisopropylchlorosilane, TEA is triethylamine, TFA is trifluoroacetic acid, THF is tetrahydrofuran, TMS is trimethylsilyl, TMSOTf is trimethylsilyl triflate, TBSOTf is t-butyldimethylsilyl triflate, tol is toluene, ts is p-toluenesulfonyl, tsOH is p-toluenesulfonic acid, TCCA is trichloroisocyanuric acid, lev is levulinyl.
The peracetylglucose, D-glucose, D-glucosamine hydrochloride in the examples below are conventional commercial products.
Example 1 Synthesis of tetraose, pentaose and Tectose molecules containing a Diaminoglucuronic acid Structure and optimization of the method for synthesizing pentasaccharide
1. Inverse synthetic analysis of tetraose 2-3, pentaose 2-4, and decaose 2-5
For the synthesis of the target compounds 2-3, 2-4 and 2-5, the invention adopts a convergent synthesis strategy, and simultaneously explores a one-pot synthesis strategy. By inverse synthetic analysis of the decasaccharide 2-5, it was found that the decasaccharide 2-5 was deprotected from the fully protected decasaccharide 2-24, whereas the decasaccharide 2-24 was obtained by regio-and stereoselective glycosylation of the pentasaccharide acceptor 2-22 with the pentasaccharide donor 2-23. The target molecule 2-4 can be obtained by one-step hydrogenation of pentasaccharide 2-22, and the pentasaccharide is obtained by removing levulinyl protection after [2+1+2] glycosylation of disaccharide receptor 2-19, monosaccharide donor 2-7 and disaccharide donor 2-21. Synthesis of pentasaccharide donors 2-23 potential donors were likewise obtained from [2+1+2] glycosidation in which the anomeric position of the disaccharide acceptor used was protected by an orthogonally removable TBS followed by removal of the TBS protecting group and reaction with N-phenyl trifluoroacetyl chloride to give the trifluoroacetyl imine ester donor. Synthesis of the tetraose target molecule 2-3 is obtained by [2+2] glycosylation of disaccharide acceptor 2-14 and disaccharide donor 2-17. The method specifically comprises the following steps:
2. Synthesis of tetraose 2-3
2.1 Synthesis of trisaccharide 2-39
First, the present invention selectively protects the C6-hydroxy group with Bz by optimizing the reaction conditions: pyridine is used as solvent, and compound 2-19 reacts with 2 equivalents of benzoyl chloride at 30 ℃ to synthesize disaccharide 2-14, and the yield is 88%.
Then, after obtaining disaccharide receptor 2-14, the present invention uses trifluoroacetyl imine ester donor 2-40 to obtain trisaccharide product 2-39 with 94% yield under optimal conditions, with selectivity β: α=4:1. The synthesis of compounds 2-39 was as follows:
2.2 Synthesis of tetraose 2-42
The product prepared by the above step is a mixture of trisaccharide 2-39 and alpha-configuration 2-39a thereof, and the mixture is used for removing Nap protecting groups by DDQ to obtain a completely separated beta-configuration product 2-41. The trisaccharide acceptor 2-41 and the monosaccharide donor 2-17 are subjected to glycosylation reaction, and the mixture of the tetrasaccharide 2-13 and the alpha-configuration 2-13a thereof is obtained, and the yield is 85%. Then, the silicon-based protecting group of the tetrasaccharide mixture is removed by HF.Py, and the beta-configuration product 2-42 with the yield of 82% is obtained.
2.3 Synthesis of tetraose 2-3
After the tetraose 2-42 is obtained, the synthesis of the target molecule 2-3 can be completed only by deprotection. Compounds 2 to 42 are first reacted with 1M aqueous lithium hydroxide solution and the resulting intermediate is then taken up in Pd (OH) 2 and/C catalytic normal pressure hydrogenation is carried out for 5 days, and finally, the target molecule 2-3 is obtained in higher yield.
The dosage proportion of each raw material in the synthesis process of the tetraose 2-3 and the specific steps are as follows:
compound S13
Compound S12 (1.81 g,5 mmol) was dissolved in 40mL tolueneDibutyl tin oxide (2.49 mg,10mmol,2 eq) was added at room temperature, the mixture was refluxed in an oil bath at 120 ℃ for 6 hours, cooled naturally for 20 minutes, tetrabutylammonium bromide (1.61 g,5mmol,1 eq) and 4-methoxybenzyl chloride (1.7 ml,12.5mmol,2.5 eq) were added rapidly, the oil bath at 115 ℃ was continued to heat and react, the mixture was kept in a slightly boiling state, stirred for 5 hours, quenched with triethylamine, concentrated under reduced pressure to remove the solvent, and the mixture was separated by column chromatography (petroleum ether: ethyl acetate 8:1-1:1) to give S13 (2.03 g,3.35mmol, 67%) as a white solid.
Wherein the compound S12 is prepared by the method described in the literature (Zhang Q, gimeno A, santana D, wang Z, values-Balbin Y, rodrii-guez-Noda LM, hansen T, kong L, shen M, overkleeft HS, verez-Bencomo V, van der Marel GA, jimenez-Barbero J, chiodo F, cod e JDC.synthetic, zwitterionic Sp1 Oligosaccharides Adopt a Helical Structure Crucial for Antibody interface. ACS centSci.2019 Aug28;5 (8): 1407-1416.).
Compound S14
Under the protection of nitrogen, compound S13 (1.87 g,3.1 mmol) is dissolved in 18mL of pyridine, 4-dimethylaminopyridine (76 mg,0.6mmol,0.2 eq) is added at room temperature, acetic anhydride (0.43 mL,4.6mmol,1.5 eq) is added after stirring for 5 minutes, the reaction is stirred at room temperature, TLC monitors the exhaustion of the reaction raw materials, ethyl acetate is added for dilution and transfer, and the mixture is washed twice with 1N HCl solution in turn, saturated NaHCO 3 Washing the solution and saturated saline solution once, and then using anhydrous Na 2 SO 4 And (5) drying. Filtering and concentrating under reduced pressure. Column chromatography (petroleum ether: ethyl acetate 8:1-2:1) afforded S14 (1.84 g,2.85mmol, 92%) as a white solid.
Compound S16
Compound S14 (1.94 g,3 mmol) was dissolved in 22mL of a mixed solvent of acetone and water in a volume ratio of 10:1, NBS (1.6 g,9mmol,3 eq) was added in two portions in an ice-water bath, the reaction was slowly warmed to room temperature and stirred for 1 hour, TLC monitored the exhaustion of the reaction materials, quenched by the addition of saturated sodium thiosulfate solution, diluted and turned out with DCM, and water and saturated NaHCO were sequentially added 3 Solution, saturated saline water washing, anhydrous Na 2 SO 4 Drying, filtering, and concentrating under reduced pressure. Column chromatography (petroleum ether: ethyl acetate 5:1-1:1) gave colorless syrup S15 (1.52 g,2.76mmol, 92%). Compound S15 (1.05 g,1.9 mmol) was dissolved in 12mL of acetone, potassium carbonate (320 mg,2.3mmol,1.2 eq) was added, stirred at room temperature for 10min, PTFAI (0.46 mL,2.8mmol,1.5 eq) was added, reacted at room temperature for 5 hours, triethylamine was added, filtered off with suction, concentrated under reduced pressure, and separated by column chromatography (petroleum ether: ethyl acetate 16:1-5:1) to give colorless syrup S16 (1.24 g,1.71mmol,90%, α: β=1.1:1).
Compound S17
Trifluoroacetyl imine ester donor S16 (100 mg,0.14mmol,1 eq) and acceptor S21 (63 mg,0.19mmol,1.4 eq) were dissolved in 2mL DCMMS dry water removal), newly activated ++>MS (200 mg), nitrogen protection, stirring at room temperature for 30min, placing the reaction at-60 ℃, adding TMSOTF (4 mu L,0.02mmol,0.15 eq), gradually heating to room temperature for reaction, stirring for 2 hours, adding triethylamine for quenching reaction, suction filtration, vacuum concentration, column chromatography separation (petroleum ether: ethyl acetate 12:1-3:1) to obtain a mixture of colorless syrups S17 and S17b (117 mg,98%, alpha: beta=1:6.5).
The receptor S21 is prepared by the method described in the literature (Noti C, de Paz J L, polio L, et al preparation and use of microarrays containing synthetic heparin oligosaccharides for the rapid analysis ofheparin-protein interactions [ J ]. Chemistry-A European Journal,2006,12 (34): 8664-8686).
Compound S8
Compound S17 (270 mg,0.31 mmol) was dissolved in a mixed solvent of 3mL of dichloromethane and methanol (2:1), sodium methoxide (4 mg,0.08mmol,0.25 eq) was added, the reaction was carried out at room temperature for 48 hours, pH was adjusted to about 7 by adding an acidic resin (monitoring with pH paper), filtration, concentration under reduced pressure, and column chromatography separation (petroleum ether: ethyl acetate 5:1-2:1) to give colorless syrup S8 (246 mg,0.3mmol, 97%).
Compound S23
Total acetylglucosamine (12 g,44 mmol) is dissolved in a mixed solvent of dichloromethane and methanol (20 mL:40 mL), potassium carbonate (3 g,22mmol,0.5 eq) is added, reacted for 4 hours at room temperature, filtered and concentrated under reduced pressure to obtain a crude product S22. The crude product S22 was dissolved in 40mL of LDMF, imidazole (11.98 g,176mmol,4 eq) was added at room temperature, dimethyl t-butylchlorosilane (14.59 g,96.8mmol,2.2 eq) was added in ice water bath, and the mixture was transferred to room temperature for reaction overnight, diluted with ethyl acetate and transferred out, and washed three times with water, saturated NaHCO in sequence 3 The solution and saturated saline were washed once each, anhydrous Na 2 SO 4 Drying, filtration, concentration under reduced pressure, column chromatography (petroleum ether: ethyl acetate 50:1-20:1) gave colorless syrup S23 (14.84 g,39.6mmol, 90%).
Compound S25
Compound S23 (9 g,24 mmol) was dissolved in 100mL tetrahydrofuran, the reaction system was placed under an ice-water bath, sodium hydride (1.92 g,48mmol,2 eq) was added in portions, and benzyl bromide (4 mL,33.6mmol,1.4 eq) was slowly added with stirring for 20min, and the reaction was gradually warmed to room temperature overnight, TLC detection of the material exhaustion, dropwise methanol quenching, filtration, concentration under reduced pressure, column chromatography separation (petroleum ether: ethyl acetate 80:1) afforded colorless syrup S24 (9.15 g,19.68mmol, 82%). Compound S24 (5.4 g,11.6 mmol) was dissolved in 35mL tetrahydrofuran, TBAF (6.6 g,23.2mmol,2 eq) was added and reacted at room temperature for 3 hours, TLC detected the starting material exhaustion, concentrated directly under reduced pressure, and separated by column chromatography (petroleum ether: ethyl acetate 5:1-1:1) to give S25 (2.74 g,10.44mmol, 90%) as a white solid.
Compound S27
Compound S25 (8.3 g,35.1 mmol) was dissolved in 330mL of DMF, imidazole (3.59 g,52.7mmol,1.5 eq) was added, diphenyltert-butylchlorosilane (10 mL,38.6mmol,1.1 eq) was added under ice-water bath, the reaction was transferred to room temperature for 24 hours, TLC was used to detect the run-up of starting material, ethyl acetate was diluted and transferred out, and three times of washing with water, saturated NaHCO were sequentially used 3 The solution and saturated saline were washed once each, anhydrous Na 2 SO 4 Drying, filtering and concentrating under reduced pressure to obtain a crude product S26. The crude product was dissolved in a mixed solvent of DCM: py (120 mL:60 mL), 4-dimethylaminopyridine (855 mg,7mmol,0.2 eq) and acetic anhydride (5 mL,52.6mmol,1.5 eq) were added, the reaction was stirred at room temperature, TLC monitored for run-up of the reaction material, dichloromethane dilution turned off, and 1N HCl solution was used twice in sequence, saturated NaHCO 3 The solution and saturated saline were washed once each, anhydrous Na 2 SO 4 And (5) drying. Filtering and concentrating under reduced pressure. Column chromatography (petroleum ether: ethyl acetate 30:1-20:1) gave colorless syrup S27 (15.6 g,30.18mmol, 86%).
Compound S35
Compound S27 (12 g,23.2 mmol) was dissolved in 250mL of dichloromethane, tetrabutylammonium nitrate (14.13 g,46.4mmol,2 eq), 2, 6-di-tert-butyl-4-methylpyridine (9.53 g,46.4mmol,2 eq) were added at room temperature, nitrogen protection, trifluoromethanesulfonic anhydride (7.8 mL,46.4mmol,2 eq) was added at-70deg.C, reaction was carried out for 30min at-70deg.C, diluted with dichloromethane and transferred out, washed with 1M HCl solution, saturated brine, anhydrous Na 2 SO 4 Drying, filtering, concentrating under reduced pressure, crystallizing and separating out generated salt in the reaction system by using ethyl glacial acetate, filtering, spin-drying, and separating by column chromatography (petroleum ether: ethyl acetate 20:1-16:1) to obtain colorless syrup S34 (9.6 g,17.17mmol, 74%). Compound S34 (3.1 g,5.5 mmol) was dissolved in 30mL of methylene chloride, 4-pyrrolidinylpyridine (163 mg,1.1mmol,0.2 eq), azido trimethylsilane (0.87 mL,6.6mmol,1.2 eq) were added and reacted at room temperature for 30min, TLC detection of the starting material exhaustion was continued with trimethylacetic acid (337 mg,3.3mmol,0.6 eq), allyl alcohol (1.75 mL,11mmol,2 eq) and the reaction system was further added, sealed, transferred to an oil bath at 35℃and reacted for 48 hours, concentrated under reduced pressure, and separated by column chromatography (petroleum ether: ethyl acetate 30:1-20:1) to give colorless syrup S35 (2.25 g,3.74mmol, 68%).
Compound S37
Compound S35 (770 mg,1.3 mmol) was dissolved in tetrahydrofuran: zinc powder (1.7 g,26mmol,20 eq) and copper sulfate (207 mg,1.3mmol,1 eq) are added to a mixed solvent of water (15 mL:1.5 mL), hydrochloric acid (0.68 mL,13mmol,10 eq) is slowly added, stirring is performed at room temperature, TLC detects the depletion of the raw material, saturated sodium bicarbonate solution is added to neutralize the hydrochloric acid, suction filtration and toluene azeotropically spin-dry the water inside to obtain a crude product, which is dissolved in 15mL of tetrahydrofuran @ MS dry), add threeEthylamine (1.4 mL,10.4mmol,8 eq), trichloroacetyl chloride (0.6 mL,5.2mmol,4 eq), reacted at room temperature for 4h, filtered, concentrated under reduced pressure, and separated by column chromatography (petroleum ether: ethyl acetate 16:1-10:1) to give colorless syrup S36 (640 mg,0.78mmol, 60%). Compound S36 (1.43 g,1.7 mmol) was dissolved in 5mL tetrahydrofuran, tetrabutylammonium fluoride (444 mg,1.7mmol,1 eq) was added and reacted at room temperature for 10 hours, TLC detected the starting material exhaustion, concentrated directly under reduced pressure, and separated by column chromatography (petroleum ether: ethyl acetate 5:1-1:1) to give S37 (0.9 g,1.5mmol, 88%) as a white foamy solid.
Compound S33
Compound S37 (1.04 g,1.7 mmol) was dissolved in dichloromethane: water: to a mixed solvent of t-butanol (2:1:1, 20 mL), TEMPO oxidant (53 mg,0.34mmol,0.2 eq), BAIB (1.1 g,3.4mmol,2 eq) were added under an ice-water bath, and the mixture was allowed to react overnight at room temperature, TLC was used to detect the run-up of the starting material, quenched with saturated sodium thiosulfate solution, diluted with dichloromethane, turned out, washed with water, the aqueous phase was stripped with dichloromethane, the organic phase was combined, saturated brine washed with anhydrous Na 2 SO 4 Drying, filtering and concentrating under reduced pressure to obtain a crude product S38. The crude product S38 was dissolved in 10mL of DMF, potassium bicarbonate (1.02 g,10.2mmol,6 eq) and benzyl bromide (0.6 mL,5.1mmol,3 eq) were added, the reaction was stirred at room temperature for 18h, diluted with ethyl acetate and turned out, washed three times with water, saturated NaHCO sequentially 3 The solution and saturated saline were washed once each, anhydrous Na 2 SO 4 Drying, filtration, concentration under reduced pressure, column chromatography (petroleum ether: ethyl acetate 12:1-8:1) gave S33 (0.84 g,1.19mmol, 70%) as a white solid.
Compound S7
Compound S33 (0.4 g,0.57 mmol) was dissolved in 18mL of a mixed solvent of methanol and methylene chloride in a volume ratio of 2:1, palladium dichloride (25 mg,0.14mmol,0.25 eq) was added, reacted at 35℃for 3 hours, suction filtered through silica gel, concentrated under reduced pressure, and separated by column chromatography (petroleum ether: ethyl acetate 5:1-1:1) to give S39 (300 mg,0.45mmol, 80%) as a white waxy solid. Compound S39 (300 mg,0.45 mmol) was dissolved in 4mL of acetone, cesium carbonate (176 mg,0.54mmol,1.2 eq) was added, stirred at room temperature for 10min, PTFAI (0.11 mL,0.68mmol,1.5 eq) was added, reacted at room temperature for 5 hours, triethylamine was added, filtered off with suction, concentrated under reduced pressure, and separated by column chromatography (petroleum ether: ethyl acetate 12:1-8:1) to give colorless syrup S7 (305 mg,0.36mmol, 81%).
Compound S75
Donor S7 (104 mg,0.12mmol,1.5 eq) and acceptor S8 (68 mg,0.083mmol,1 eq) were mixed, toluene azeotroped twice, water was dried by spinning, and dissolved in 2mLDCMMS drying Water removal x 2), adding +.>MS (200 mg), nitrogen protection, stirring at-40℃for 10min, adding TBSOTf (4. Mu.L, 0.017mmol,0.2 eq), reacting at-40℃for 2 h, quenching with triethylamine, suction filtration, concentrating under reduced pressure, flash column chromatography (Petroleum ether: ethyl acetate 8:1-2:1) to give colorless clear syrup S75 (106 mg,0.072mmol, 87%).
Compounds 2-19
Compound S75 (180 mg,0.12mmol,1 eq) was dissolved in 5mL of dichloromethane, TFA (0.25 mL) was added under ice-water bath, gradually warmed to room temperature for reaction, TLC monitored for material exhaustion, the reaction system was poured into ice saturated sodium bicarbonate solution, dichloromethane extracted twice, the organic phases combined, saturated brine washed, no waterWater Na 2 SO 4 Drying, filtration, concentration under reduced pressure, column chromatography (petroleum ether: ethyl acetate 5:1-1:1) gave colorless syrup 2-19 (138 mg,0.11mmol, 94%).
Compounds 2-14
Compound 2-19 (555 mg, 0.4573 mmol) was dissolved in 14mL of dry pyridine, nitrogen-protected, bzCl (0.11 mL,0.906mmol,2 eq) was added, and reacted in an oil bath at 30℃after 0.5H, the TCL assay starting material reacted, diluted with ethyl acetate, and the reaction mixture was quenched with H 2 Washing with O and saturated NaCl solution, anhydrous Na 2 SO 4 Drying, concentrating, dry loading, and column chromatography separation and purification (petroleum ether: ethyl acetate=10:1→2:1) gave foam solid 2-14 (53 mg,0.401mmol, 88%).
Compounds 2-40
Compound 2-12 (400 mg,0.53 mmol) was dissolved in 4mL of acetone and 1mL of H was added 2 O is placed in 0 ℃ and NaHCO is alternately added 3 (224 mg,2.66mmol,5 eq) and TCCA (130 mg,0.559mmol,1.05 eq) were reacted, after 2h the TCL detected complete reaction of the starting material, saturated Na was added 2 S 2 O 3 Quenching the solution, concentrating, dissolving in dichloromethane, diluting, and adding H 2 Washing with O and saturated NaCl solution, anhydrous Na 2 SO 4 Drying, concentrating, stirring by dry method, and separating and purifying by column chromatography to obtain 298mg of intermediate without thioglycoside. Dissolving the intermediate in 6mL of acetone, and adding Cs 2 CO 3 (226 mg,0.693mmol,1.3 eq) in ice bath with PTFAI (0.11 mL,0.693mmol,1.3 eq) at room temperature, 2h after Et 3 N, filtration, concentration, dry sample stirring, column chromatography separation and purification (petroleum ether: ethyl acetate=20:1→10:1) gave 2-40 (304 mg,0.373mmol, 70%) as a pale yellow foamy solid.
Wherein, the compounds 2-12 are prepared by a method in literature (Hui Liu, zhi-Fen Liang, han-Jian Liu, jin-Xi Liao, li-Jun Zhong, yuan-Hong Tu, qing-Ju Zhang, bin Xiong, and Jian-Song Sun Journal of the American Chemical Society 2023 145 (6), 3682-3695.).
Compounds 2-39
The acceptor 2-14 (41 mg,0.0308 mmol) and the donor 2-40 (50 mg,0.0616mmol,2 eq) were weighed into a reaction flask, azeotroped 3 times with toluene, pumped with oil for 2h, and the activated was addedMS, nitrogen aeration 3 times, charging 2mL of dried DCM into the system, stirring at 0deg.C for 10min, charging TBSOTf (1.4 uL,0.00616mmol,0.2 eq), 4h later, TCL detecting receptor reaction completely, adding Et 3 N-quenching, filtration, concentration, dry-mixing, column chromatography (toluene: ethyl acetate=50:1→10:1) gave a mixture of foamy solids 2-39 and 2-39a (57 mg,0.0291mmol,94%, β/α=4:1).
Compounds 2-41
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A mixture of compounds 2-39 and 2-39a (4476 mg,0.228 mmol) was dissolved in 8mL of methylene chloride, 1.6mL of Buffer solution was added, DDQ (207 mg,0.912mmol,4 eq) was added in small portions at 0deg.C, and the mixture was gradually heated to 10deg.C to react, and saturated Na was added after 4.5h 2 S 2 O 3 Quenching the solution, filtering, and using H 2 O is washed twice, saturated NaCl solution is washed once, anhydrous Na 2 SO 4 Drying, concentrating, dry-mixing, and column chromatography (petroleum ether: ethyl acetate=10:1→2:1) gave foam solid 2-41 (219 mg,0.121mmol, 53%).
Compounds 2-17
Compound S44 (5.9 g,13.09 mmol) was dissolved in 60mL of dichloromethane, DMAP (0.64 g,5.24mmol,0.4 eq), triethylamine (7.3 mL,52.36mmol,4 eq) were added, bzCl (3.1 mL,26.18mmol,2 eq) was slowly added under nitrogen blanket, stirred for 4h, concentrated under reduced pressure, and isolated by column chromatography (petroleum ether: ethyl acetate 10:1-5:1) to give S76 (5.66 g,10.21mmol, 78%) as a white solid. Compound S76 (1.28 g,2.3 mmol) was placed in a 50mL round bottom flask, borane tetrahydrofuran solution (14 mL,14mmol,6 eq) was added, stirred for 10min under ice-water bath, copper triflate (42 mg,0.12mmol,0.05 eq) was added, gradually warmed to room temperature, stirred overnight, quenched with methanol dropwise, filtered under suction, concentrated under reduced pressure, and separated by column chromatography (petroleum ether: ethyl acetate 12:1-5:1) to give white solid S-77 (1.2 g,2.16mmol, 94%).
Wherein, the compound S44 is prepared by a method in literature (Shie C R, tzeng Z H, kulkarni S, et al Cu (OTf) 2as an Efficient and Dual-Purpose Catalyst in the Regioselective Reductive Ring Opening of Benzylidene Acetals [ J ]. Angewandte Chemie International Edition,2005,44 (11): 1665-1668.).
Compound S77 (5.35 g,9.61 mmol) was dissolved in 5.5mL DMF, imidazole (0.98 g,14.42mmol,1.5 eq), DMAP (0.12 g,0.96mmol,0.1 eq) and TIPSCl (2.5 mL,11.68mmol,1.2 eq) were added under ice water bath, transferred to room temperature for 3h, ethyl acetate was diluted and transferred out, and three times water washes were performed sequentially with saturated NaHCO 3 The solution and saturated saline were washed once each, anhydrous Na 2 SO 4 Drying, filtration, concentration under reduced pressure, column chromatography (petroleum ether: ethyl acetate 100:1-50:1) gave colorless syrup S78 (6.28 g,8.17mmol, 85%). Compound S78 (6.28 g,8.18 mmol) was dissolved in 80mL of a mixed solvent of acetone and water in a volume ratio of 3:1, and TCCA (2 g,8.59mmol,1.05 eq) and NaHCO were added in two portions in an ice-water bath 3 (3.43 g,40.88mmol,5 eq) at 0deg.C for 1 hour, quenched by addition of saturated sodium thiosulfate solution, diluted with ethyl acetate and transferred out, followed by water and saturated NaHCO 3 Solution, saturated saline water washing, anhydrous Na 2 SO 4 Drying, filtering, and concentrating under reduced pressure. Column chromatography (petroleum ether: ethyl acetate 20:1-10:1) gave colorless syrup S79 (4.26 g,6.46mmol, 79%). Compound S79 (1 g,1.61 mmol) was dissolved in 10mL of acetone, cesium carbonate (630 mg,1.93mmol,1.2 eq) was added, stirred at room temperature for 10min, PTFAICl (0.4 mL,2.41mmol,1.5 eq) was added, reacted at room temperature for 5 hours, triethylamine was added, filtered off with suction, concentrated under reduced pressure, and the column chromatography was separated (petroleum ether: ethyl acetate 100:1-60:1) to give colorless syrup 2-17 (1 g,1.26mmol, 78%).
Compounds 2-13
Acceptor 2-41 (219 mg,0.121 mmol) and donor 2-17 (280 mg,0.354mmol,2.9 eq) were weighed into a reaction flask, azeotroped 3 times with toluene, pumped with oil for 2h, and activated was addedMS, nitrogen aeration 3 times, charging 6mL of dried DCM into the system, stirring at-20deg.C for 10min, charging TfOH (3.5 uL,0.0396mmol,0.33 eq), detecting complete reaction of receptor by TCL after 2h, adding Et 3 N quenching, filtering, concentrating, dry-mixing, column chromatography (Petroleum ether: ethyl acetate=20:1→5:1) to obtain a mixture of foamy solid 2-13 and 2-13a (248 mg,0.103mmol,85%, beta/alpha)>10:1)。[α] D 20 ＝+23.9(c 1.1，CHCl 3 )； 1 H NMR(400MHz,CDCl 3 )δ8.07–8.00(m,2H),7.95(d,J＝6.4Hz,2H),7.84(d,J＝8.2Hz,1H),7.56(t,J＝7.4Hz,1H),7.48(t,J＝7.4Hz,1H),7.43(d,J＝7.8Hz,2H),7.40–7.18(m,29H),7.18–7.09(m,14H),7.09–7.02(m,4H),6.95(t,J＝7.2Hz,1H),6.89(d,J＝5.9Hz,1H),5.32(t,J＝8.5Hz,1H),5.21–5.11(m,3H),5.11–4.99(m,3H),4.92(d,J＝10.5Hz,1H),4.89–4.81(m,2H),4.77(d,J＝12.5Hz,1H),4.73–4.64(m,3H),4.59(d,J＝10.9Hz,2H),4.54(s,1H),4.50–4.41(m,5H),4.37(d,J＝11.6Hz,2H),4.33–4.24(m,2H),4.16(d,J＝11.1Hz,1H),4.13–3.97(m,4H),3.96–3.70(m,9H),3.69–3.63(m,1H),3.54–3.38(m,3H),3.35–3.21(m,1H),3.10(d,J＝29.5Hz,2H),2.89(d,J＝11.9Hz,1H),1.44–1.28(m,4H),1.10–1.05(m,2H),1.05–0.94(m,21H). 13 C NMR(100MHz,CDCl 3 )δ167.54,166.03,165.14,163.47,162.58,162.50,156.63,138.67,138.01,137.93,137.86,137.75,137.60,137.23,134.96,133.19,133.00,130.11,130.08,129.86,129.58,129.15,128.74,128.58,128.52,128.49,128.45,128.35,128.32,128.28,128.26,128.23,128.02,128.00,127.96,127.91,127.87,127.80,127.77,127.72,127.53,127.34,127.21,102.10,101.14,98.83,96.46,92.40,91.74,91.66,82.92,78.65,77.77,77.54,77.26,76.60,76.47,75.84,75.51,75.44,75.29,75.22,74.95,74.87,74.16,73.81,73.04,72.83,70.07,67.68,67.46,67.15,66.90,64.74,62.23,57.92,57.28,54.63,50.53,50.18,47.12,46.15,29.12,27.92,27.47,23.36,18.06,11.90.
Compounds 2-42
A mixture of compounds 2-13 and 2-13a (248 mg,0.103 mmol) was dissolved in 3mL tetrahydrofuran and 3mL pyridine, 0.3mL HF. Py (65.9%) was added in ice bath, and the reaction was stirred at room temperature, diluted with ethyl acetate after 24h, poured into saturated NaHCO under ice bath 3 Stirring the solution for 10min, extracting, washing the saturated NaCl solution once, and anhydrous Na 2 SO 4 Drying, concentrating, dry-mixing, and column chromatography separation and purification (petroleum ether: ethyl acetate=10:1→2:1) to obtain foam solid 2-42 (191 mg,0.0845mmol, 82%), [ alpha ]]D 20 ＝+12.4(c 2.3，CHCl 3 )， 1 H NMR(400MHz,CDCl 3 )δ8.02(d,J＝7.2Hz,2H),7.95(d,J＝7.2Hz,2H),7.85(d,J＝8.4Hz,1H),7.55(t,J＝7.4Hz,1H),7.47(t,J＝7.4Hz,1H),7.41(t,J＝7.7Hz,2H),7.39–7.27(m,18H),7.26–7.09(m,27H),7.08–7.05(m,2H),7.02(t,J＝7.0Hz,1H),6.94(d,J＝6.0Hz,1H),5.35–5.27(m,1H),5.16(d,J＝7.3Hz,3H),5.07(t,J＝9.5Hz,2H),5.01(d,J＝12.1Hz,1H),4.84(dd,J＝11.1,3.9Hz,2H),4.81–4.76(m,1H),4.72(t,J＝8.8Hz,2H),4.69–4.59(m,4H),4.58(d,J＝4.2Hz,1H),4.54(s,1H),4.47(d,J＝10.5Hz,1H),4.45–4.34(m,5H),4.34–4.25(m,2H),4.22–4.05(m,3H),3.98(d,J＝9.2Hz,1H),3.94–3.80(m,6H),3.79–3.70(m,4H),3.66(dd,J＝10.0,3.3Hz,1H),3.62–3.53(m,1H),3.49–3.37(m,2H),3.36–3.22(m,1H),3.18–3.00(m,2H),2.99–2.82(m,1H),1.44–1.28(m,4H),1.13–0.95(m,2H). 13 C NMR(100MHz,CDCl 3 )δ167.59,166.03,165.24,163.31,162.58,162.49,156.67,138.69,137.93,137.77,137.73,137.69,137.53,137.18,134.95,133.31,133.01,130.11,129.66,129.58,129.27,128.73,128.59,128.53,128.49,128.39,128.36,128.32,128.29,128.10,128.04,128.00,127.94,127.89,127.84,127.77,127.64,127.34,127.21,101.94,101.12,99.14,96.53,92.39,91.82,91.76,82.46,78.58,77.46,77.28,75.86,75.54,75.48,75.34,75.31,75.01,74.82,74.72,74.60,74.35,73.92,73.00,72.92,69.40,67.71,67.51,67.17,66.94,64.69,61.63,57.73,57.12,54.58,50.55,50.20,47.12,46.11,29.11,27.85,27.44,23.36.
Compounds 2-3
Dissolving compound 2-42 (69 mg,0.0305 mmol) in 6mL tetrahydrofuran and 2mL methanol, adding 0.2mL1M LiOH aqueous solution, reacting at room temperature, neutralizing with acid resin to weak acidity after 11H, quenching, filtering, concentrating under reduced pressure, dry stirring, separating and purifying by column chromatography to obtain intermediate with Bz protecting group removed, dissolving the intermediate in 1mL tetrahydrofuran, adding 5mL tertiary butanol and 2mL H 2 O, 0.05mL AcOH, 120mg Pd (OH) 2 C (20%) under ice bath H 2 Aeration 15min, H of 1atm 2 Reacting at room temperature for 5 days under ambient condition, filtering, concentrating under reduced pressure, and purifying with Sephadex LH-20 column (CH 3 OH:H 2 O=1:1) to give compound 2-3 (23 mg,0.0259mmol, 85%). 1 H NMR(600MHz,D 2 O)δ5.03(d,J＝8.4Hz,1H),4.88(d,J＝3.9Hz,1H),4.63(d,J＝8.5Hz,1H),4.51(d,J＝7.9Hz,1H),4.39(d,J＝2.5Hz,1H),4.20(d,J＝3.1Hz,1H),4.11(dd,J＝10.8,8.6Hz,1H),4.04(dd,J＝11.0,10.0Hz,1H),3.93–3.84(m,5H),3.84–3.77(m,4H),3.76–3.68(m,4H),3.64(dd,J＝11.8,6.7Hz,1H),3.59(t,J＝9.8Hz,1H),3.56–3.52(m,1H),3.48–3.44(m,1H),3.44–3.39(m,2H),3.34–3.30(m,1H),3.01(t,J＝7.6Hz,2H),2.05–1.94(m,9H),1.73–1.63(m,4H),1.50–1.41(m,2H). 13 C NMR(150MHz,D 2 O)δ175.78,174.67,174.59,104.36,103.62,101.23,98.44,80.23,80.17,76.19,75.69,75.56,75.51,74.66,72.83,70.34,69.84,69.37,67.97,67.86,67.21,61.15,60.44,60.16,54.57,53.54,51.49,39.33,28.02,26.41,22.32,22.31,22.12,21.96.
3. Synthesis of pentasaccharide 2-4 and decasaccharide 2-5
3.1 Synthesis of Compounds 2-18, 2-16, 2-21
In the process of synthesizing the pentasaccharide 2-4, the invention adopts the strategy of [3+2] to carry out glycosylation to synthesize the pentasaccharide. The synthesis of the monosaccharide donors and disaccharide donors involved therein is shown below.
Synthesis of Compounds 2-18: starting from D-glucose, compounds 2-54 were obtained in 60% yield. Then, BH of 1M is used 3 THF was reduced under TMSOTF catalysis to cleave the benzylidene protecting group allowing bare leakage of C6-hydroxy, giving compound 2-55 in 95% yield. The two hydroxyl groups of the bare leak were protected with Lev, yielding compounds 2-56 almost quantitatively. Finally, NBS is used for hydrolyzing the anomeric thioglycoside, and then the obtained product is reacted with PTFAI to prepare the trifluoroacetyl imine ester donor 2-18.
Synthesis of Compounds 2-16: using the compound 2-12 as a starting material, the anomeric position of 2-12 was converted to allyl, and the compound 2-63 was obtained in 80% yield. Finally removing the naphthylmethyl protection by using DDQ to obtain the galactosamine receptor 2-16.
Synthesis of Compounds 2-21: beta-configuration acceptor 2-16 is reacted with glucose donor 2-18 to obtain single beta-configuration disaccharide 2-66 in 89% yield. Then palladium chloride is used to remove allyl at the anomeric position, then alkali and PTFAI are stirred for 10min and then added into a substrate to react, thus preparing the trifluoroacetyl imine ester donor 2-21, and the two-step yield is 48%.
3.2 Synthesis of pentasaccharide 2-70, 2-69
In the process of synthesizing 2-70 and 2-69, tfOH is used as an accelerator, and the compound 2-19 or 2-20 and the monosaccharide donor 2-7 react at the temperature of minus 60 ℃ to obtain trisaccharides 2-72 and 2-49 with single configuration. It was further attempted to construct beta-glycosidic linkages by coupling 2-72 or 2-49 to 2-21 using methods that would allow efficient construction of 1, 2-trans glycosidic linkages, but with lower stereoselectivity.
In order to increase the synthesis efficiency of pentasaccharide (2-70, 2-69) and further to achieve efficient synthesis of the surface saccharide antigen (decasaccharide) of acinetobacter baumanii ATCC 17961, it is necessary to increase the stereoselectivity of [2+3] glycosylation in pentasaccharide synthesis. Since another sugar chain is present at the C4-O-position, stereoselective glycosylation at galactose C3-OH is critical.
Thus, the present invention employs a method of constructing beta glycosidic linkages through levulinyl (Lev) remote hydrogen bond induction of adjacent saccharides. The method realizes the stereoselective synthesis of [2+3] glycosylation through the participation of adjacent glycosyl induced by Lev remote hydrogen bond, and solves the stereoselective problem of high steric hindrance and low reactivity receptor. Based on the above, the invention synthesizes 2-72 and 2-49 from disaccharides 2-19 and 2-20 with 1, 2-cis-glycosidic bond construction, and synthesizes pentasaccharide receptors 2-70 and 2-69. The synthesis reaction general formula of the pentasaccharide receptors 2-70 and 2-69 is as follows:
wherein, the disaccharide donor 2-21 has the structural formula:
the structural formulas of the receptors 2-72 and 2-49 are as follows:
by optimizing the reaction conditions, the optimum reaction conditions are shown in the following table 1:
TABLE 1
The reaction was optimized to give the desired pentasaccharide 2-69 in 85% yield with higher stereoselectivity (β: α=4.5:1) by glycosylation of disaccharide donor 2-21 with acceptor 2-49. And the disaccharide donor 2-21 is coupled with the trisaccharide receptor 2-72, so that the pentasaccharide 2-70 can be generated in 81 percent yield and has good stereoselectivity (beta: alpha=5:1). Among these, the advantage of the compound 2-21 in terms of glycosylation stereoselectivity is benefited by the fact that the protecting group in the compound 2-21 is C2',6' -di-O-acetopropenyl (Lev).
Finally, the general experimental procedure for the stereoselective glycosylation involving the distal Lev protecting group during synthesis of pentasaccharide receptors 2-70, 2-69 was as follows:
the mixture of donor (2 equiv), acceptor (1 equiv) was co-evaporated three times with toluene and then dried with fresh flameMolecular sieve and dried DCM (0.023M) were added sequentially to a round bottom flask. After stirring at room temperature for 10 min under nitrogen, and stirring at 0deg.C for 5 min, TMSOTF (0.2 eq) was added and the reaction was gradually warmed to room temperature until TLC analysis indicated complete consumption of the acceptor. With saturated NaHCO 3 The reaction was quenched with water, filtered under vacuum, and concentrated under reduced pressure. The crude product was purified by gel column chromatography (DCM: meoh=1:1) followed by silica gel column chromatography to give the corresponding product.
Accordingly, the synthesis method of the compounds 2-69 after optimization is as follows:
following a general experimental procedure for stereoselective glycosylation with participation of the distal Lev protecting group, starting material donor 2-21 (166 mg,0.137 mmol), acceptor 2-49 (110 mg,0.068 mmol), TMSOTf (2.5 μl,0.037 mmol) and DCM (3 mL) were reacted in a 10mL round bottom flask gradually warmed to room temperature by ice water bath for 4 hours, the crude product was purified by gel column chromatography (DCM: meoh=1:1) and then isolated and purified by silica gel column chromatography to give white foamy solid 2-69 (152 mg,85%, α: β=1:4.5, pe: ea=3:1, rf=0.4).
The synthesis method of the compound 2-70 after optimization is as follows:
following a general experimental procedure for stereoselective glycosylation with the participation of the distal Lev protecting group, starting material donor 2-21 (243 mg,0.2 mmol), acceptor 2-72 (180 mg,0.1 mmol), TMSOTf (3.5 μl,0.02 mmol) and DCM (4 mL) were reacted in a 10mL round bottom flask gradually warmed to room temperature by ice water bath for 4 hours, the crude product was purified by gel column chromatography (DCM: meoh=1:1) and then isolated and purified by silica gel column chromatography to give white foamy solid 2-70 (230 mg,81%, α: β=1:5, pe: ea=2.5:1, rf=0.25).
3.3 Synthesis of pentasaccharide 2-4
During the synthesis of 2-70, a mixture of two configurations 2-70 and 2-70a is obtained, and the single-configuration product is obtained after removing part of protecting groups. Then, two Lev protecting groups on the pentasaccharide were removed using an acetic acid solution of hydrazine hydrate, and pentasaccharide receptor 2-22 was obtained in high yield of 91%. Finally, the pentasaccharide 2-22 is hydrogenated in one step to obtain the target molecule 2-4.
3.4 Synthesis of pentasaccharide 2-23
Pentasaccharide 2-23 was synthesized using a [3+1+1] glycosylation synthesis strategy. After synthesis of 2-69, TBAF was used to remove the exocephalic TBS protecting group of compound 2-69, and then reacted with PTFAI to produce trifluoroacetyl imine ester donor 2-23 with a two-step yield of 84%.
3.5 Synthesis of decasaccharide 2-5
With the pentasaccharide acceptor 2-22 and pentasaccharide donor 2-23, the strategy of [5+5] was used to synthesize the decasaccharide 2-24, and experiments have found that the regioselective glycosylation of [5+5] can occur at the C-6 position of the pentasaccharide acceptor. For the construction of 1, 2-cis-glycosidic bond in the synthesis of decasaccharide, the invention adopts additive control or solvent effect to complete the synthesis of alpha-glycoside. Through screening the [5+5] glycosylation condition, the reaction is finally carried out at 20 ℃, TBSOTf is selected as a catalyst, the alpha selectivity of the reaction can be improved, and meanwhile, the reaction yield can be obviously improved by improving the temperature.
After obtaining the decasaccharide product 2-24, the two configuration products 2-75 and 2-76 were separated by removing the two Lev protecting groups on the decasaccharide using an acetic acid solution of hydrazine hydrate. With compounds 2-22 and 2-75, the synthesis of the target molecule is completed by final hydrogenation. In Pd (OH) 2 and/C as catalyst, and hydrogenating at normal pressure for 5 days to obtain target molecule 2-5 in 71% yield.
The dosage proportion of each raw material in the synthesis process of the pentasaccharide 2-4 and the decasaccharide 2-5 and the specific steps are as follows:
compounds 2-55
Compound 2-54 (6.345 g,13.6 mmol) was dissolved in 64mL dry dichloromethane and added 1M BH under nitrogen protection in an ice bath 3 THF (27 mL,27.2mmol,2 eq), TMSOTF (0.25 mL,1.36mmol,0.1 eq), gradually warmed to RT, and after 4h TCL detects complete reaction of the starting material, et is added under ice bath 3 N, meOH the reaction was quenched, concentrated, and wet loaded, and separated by column chromatography (petroleum ether: ethyl acetate=10:1→2:1) to give 2-55 (6.04 g,12.9mmol, 95%) as a white solid.
Compounds 2-54 were prepared using the methods described in the literature (Wang, CC., lee, JC., luo, SY. Et al, regioselective one-pot protection of carbohydrates. Nature 446,896-899 (2007)).
Compounds 2-56
Compounds 2-55 (4.94 g,10.6 mmol), EDCI (8.128 g,42.4mmol,4 eq), DMAP (647 mg,5.3mmol,0.5 eq) were dissolved in 50mL dry dichloromethane, under nitrogen protection, levOH (4.3 mL,42.4mmol,4 eq), DIPEA (7 mL,42.4mmol,4 eq) were added under ice bath and gradually warmed to room temperature for reaction, after 3.5h TCL detected complete reaction of the starting material,diluting with dichloromethane, and adding H 2 Washing with O and saturated NaCl solution, anhydrous Na 2 SO 4 Drying, concentrating, dry-mixing, and column chromatography (petroleum ether: ethyl acetate=10:1→2:1) gave white solid 2-56 (6.95 g,10.5mmol, 99%).
Compounds 2-18
Compound 2-56 (4.12 g,6.22 mmol) was dissolved in 40mL of acetone and 4mL of H was added 2 Adding NBS (3.321 g,18.7mmol,3 eq) in ice bath, reacting at room temperature, adding NBS (2.214 g,12.4mmol,2 eq) after 1h, continuing to react until the raw materials are reacted, adding saturated Na 2 S 2 O 3 Quenching the solution, concentrating, dissolving in dichloromethane, diluting, and adding H 2 Washing with O and saturated NaCl solution, anhydrous Na 2 SO 4 Drying, concentrating, stirring by a dry method, and separating and purifying by column chromatography to obtain 3.224g of intermediate with the thioglycoside removed. The intermediate was dissolved in 32mL of acetone and Cs was added 2 CO 3 (2.8238 g,8.68mmol,1.4 eq) and PTFAI (1.3 mL,8.68mmol,1.4 eq) were added in ice bath and reacted at room temperature after 2h Et was added 3 N, filtration, concentration, dry-mixing, column chromatography separation and purification (petroleum ether: ethyl acetate=10:1→3:1) gave 2-18 (3.712 g,5.10mmol, 82%) as a tan oil.
Compounds 2-63
Compounds 2 to 12 (110 mg,0.15mmol,1 eq) were reacted under nitrogen,MS (500 mg) was dissolved in 5mL DCM (+.>MS drying), after stirring at 0deg.C for 10min, allyl alcohol (0.1 mL,1.46 mmol) was added10 eq), NIS (46 mg,0.2mmol,1.4 eq), triflic acid (8 μl,0.09mmol,0.6 eq), stirring overnight at 0 ℃ for 6h, quenching with triethylamine, saturated sodium thiosulfate solution, removing molecular sieves by suction filtration, concentrating under reduced pressure, separating by column chromatography (petroleum ether: ethyl acetate 16:1-10:1) to give colorless syrup 2-63 (73 mg,0.12mmol, 80%).
Compounds 2-16
Compound 2-63 (1.902 g,2.78 mmol) was dissolved in 25.5mL of dichloromethane, 2.5mL of Buffer solution was added, DDQ (945 mg,4.16mmol,1.5 eq) was added in small portions under ice bath, gradually warmed to room temperature and reacted overnight, TCL was detected that the starting material was completely reacted, saturated Na was added 2 S 2 O 3 Quenching the solution, filtering, and using H 2 O is washed twice, saturated NaCl solution is washed once, anhydrous Na 2 SO 4 Drying, concentrating, dry-mixing, and column chromatography separation and purification (petroleum ether: ethyl acetate=10:1→4:1) gave foam-like solid 2-16 (1.237 g,2.27mmol, 82%).
Compounds 2-66
The acceptor 2-16 (327 mg,0.6 mmol) and the donor 2-18 (660 mg,0.9mmol,1.5 eq) were weighed into a reaction flask, azeotroped 3 times with toluene, pumped with oil for 2h, and added to the activatedMS, purging with nitrogen for 3 times, charging 12mL of dry DCM into the system, stirring at-20deg.C for 10min, charging TBSOTf (28 uL,0.12mmol,0.2 eq), and adding saturated NaHCO after 3h 3 The solution was quenched, filtered, concentrated, dry-mixed, and separated by column chromatography (petroleum ether: ethyl acetate=10:1→3:1) to give foam solid 2-66 (578 mg,0.53 mmol, 89%).
Compounds 2-21
Compound 2-66 (1.194 g,1.10 mmol) was dissolved in 15mL dichloromethane and 45mL methanol and PdCl was added 2 (49 mg,0.275mmol,0.25 eq) in 35 ℃ oil bath, filtering with celite and silica gel after 3h, dry column chromatography to obtain 0.937g intermediate; cs is processed by 2 CO 3 (440mg，1.35mmol，1.2eq)、Et 3 N (0.19 mL,1.35mmol,1.2 eq), PTFAI (0.37 mL,2.44mmol,2.2 eq) and 5mL acetone are added into a reaction bottle, nitrogen protection is carried out, stirring is carried out at room temperature for 5min, the intermediate is dissolved in 15mL acetone under ice bath, the mixture is added into the bottle, the mixture is gradually warmed to room temperature for reaction, et is added after 3h 3 N, filtration, concentration, dry sample stirring, column chromatography separation and purification (Petroleum ether: ethyl acetate=15:1→4:1) gave foam solid 2-21 (640 mg,0.53mmol, 48%).
Compounds 2-7
Compound 2-36 (2.07 g,2.95 mmol) was dissolved in 20mL of acetone and 2mL of H was added 2 O, adding NBS (2.102 g,11.8mmol,4 eq) in ice bath, reacting at room temperature, adding NBS (526 mg,2.95mmol,1 eq) after 2h, continuing to react until the raw material is reacted, adding saturated Na 2 S 2 O 3 Quenching the solution, concentrating, dissolving in dichloromethane, diluting, and adding H 2 Washing with O and saturated NaCl solution, anhydrous Na 2 SO 4 Drying, concentrating, dry mixing, and column chromatography separation and purification to obtain 1.594g of intermediate with removed thioglycoside. Dissolving the intermediate in 16mL of acetone, and adding Cs 2 CO 3 (1.013 g,4.43mmol,1.5 eq) and PTFAI (0.8 mL,5.31mmol,1.8 eq) were added in ice bath and reacted at room temperature after 2h Et was added 3 N, filtration, concentration, dry-mixing, column chromatography separation and purification (petroleum ether: ethyl acetate=15:1→8:1) gave 2-7 (1.762 g,2.30mmol, 78%) as a pale yellow waxy solid.
Compounds 2 to 36 were prepared by the methods described in the literature (Zhang, yonglian; knapp, spencer. Simple beta-glycosylation of peptides (Article) [ J ]. Tetrahedron,2018, vol.74 (23): 2891-2903.).
Compounds 2-72
Acceptor 2-19 (958 mg,0.782 mmol) and donor 2-7 (810 mg,1.06mmol,1.35 eq) were weighed into a reaction flask, azeotroped 3 times with toluene, pumped with oil for 2h, and activated was addedMS, nitrogen aeration 3 times, charging 48mL of dry DCM into the system, placing at-60deg.C, stirring for 10min, charging TfOH (7 uL,0.0782mmol,0.1 eq), detecting complete reaction of receptor by TCL after 4h, adding Et 3 N-quenching, filtration, concentration, dry-mixing, column chromatography (petroleum ether: ethyl acetate=10:1→3:1) gave foam solid 2-72 (1.255 g,0.695mmol, 89%).
Compounds 2-70
Acceptor 2-72 (180 mg,0.1 mmol) and donor 2-21 (243 mg,0.2mmol,2 eq) were weighed into a reaction flask, azeotroped 3 times with toluene, pumped with oil for 2h, and added to the activatedMS, nitrogen purging 3 times, charging 4mL of dried DCM into the system, stirring at 0deg.C for 10min, charging TMSOTF (3.5 uL,0.02mmol,0.2 eq), and adding saturated NaHCO after 4h 3 The solution was quenched, filtered and concentrated, separated using a gel column (MeOH: dcm=1:1), then dry loaded, and column chromatographed (petroleum ether: ethyl acetate=10:1→2.5:1) to give a mixture of foamy solids 2-70 and 2-70a (230 mg,0.0813mmol,81%, β/α=5:1). [ alpha ]]D 20 ＝+2.2(c 0.55，CHCl 3 )； 1 H NMR(400MHz,CDCl 3 )δ8.47(dd,J＝15.9,5.6Hz,1H),7.92(d,J＝8.1Hz,1H),7.53(d,J＝7.9Hz,1H),7.36(d,J＝5.5Hz,4H),7.34–7.29(m,12H),7.28–7.23(m,17H),7.18(s,21H),7.12–7.04(m,5H),6.96(d,J＝9.2Hz,1H),6.86(d,J＝6.4Hz,1H),5.31–5.23(m,1H),5.22–5.10(m,4H),5.08–4.99(m,2H),4.88–4.77(m,4H),4.76–4.70(m,4H),4.69–4.62(m,4H),4.57(d,J＝11.0Hz,2H),4.53–4.45(m,5H),4.40(d,J＝10.4Hz,5H),4.35–4.18(m,4H),4.09–3.95(m,4H),3.94–3.87(m,4H),3.86–3.84(m,1H),3.84–3.78(m,5H),3.75–3.67(m,1H),3.64–3.58(m,2H),3.57–3.49(m,3H),3.45(d,J＝9.8Hz,1H),3.42–3.31(m,1H),3.28–3.14(m,2H),3.09–2.83(m,2H),2.72–2.62(m,1H),2.58(t,J＝6.2Hz,2H),2.48–2.33(m,3H),2.15(s,5H),2.04(s,3H),1.57–1.39(m,4H),1.25–1.13(m,2H). 13 C NMR(100MHz,CDCl 3 )δ208.18,206.72,172.26,171.58,168.00,163.79,162.64,162.32,161.90,156.25,139.61,139.12,138.59,138.20,138.04,138.01,137.53,137.38,137.11,136.67,135.16,129.18,128.61,128.58,128.47,128.34,128.26,128.23,128.11,127.93,127.85,127.83,127.78,127.70,127.62,127.57,127.44,127.32,127.26,101.91,101.46,99.64,99.23,97.55,92.83,92.52,92.40,91.76,82.32,80.97,79.03,78.37,77.91,77.69,76.32,76.17,75.61,75.29,75.13,75.02,74.88,74.71,74.65,73.76,73.55,73.30,73.09,72.77,69.93,69.08,68.47,67.97,67.43,67.22,62.80,57.66,57.08,55.96,54.72,50.60,50.18,47.45,46.41,37.90,37.79,30.04,29.84,29.42,28.15,27.87,27.76,27.40,23.65.
Compounds 2-22
Compound 2-70 (319 mg,0.148 mmol) was dissolved in 3.2mL of dichloromethane, 1.6mL of pyridine, 1mL of acetic acid were added sequentially with stirring, and 0.15mL of N was added with ice bath 2 H 4 ·H 2 O/AcOH (volume ratio 1:1), after 5.5h at room temperature, dichloromethane was diluted, washed with 1M hydrochloric acid and then saturated NaHCO 3 Washing with solution and NaCl solution, anhydrous Na 2 SO 4 Drying and concentratingThe mixture was concentrated, dried, and subjected to column chromatography (petroleum ether: ethyl acetate=5:1→1.5:1) to give 2-22 (356 mg,0.135mmol, 91%) as a foamy solid. 1 H NMR(400MHz,d 6 -Acetone)δ8.49(d,J＝9.3Hz,1H),8.41(t,J＝9.5Hz,1H),8.23(d,J＝7.6Hz,1H),8.01(d,J＝8.9Hz,1H),7.42(dd,J＝11.9,7.4Hz,6H),7.34–7.29(m,20H),7.28–7.24(m,17H),7.23(s,5H),7.21(s,8H),7.20(s,3H),7.10(t,J＝7.3Hz,1H),5.56(d,J＝8.4Hz,1H),5.19(q,J＝12.6Hz,4H),5.03(t,J＝10.2Hz,2H),4.93–4.76(m,8H),4.75–4.61(m,6H),4.60–4.50(m,7H),4.46–4.34(m,6H),4.29–4.20(m,3H),4.16(d,J＝2.7Hz,1H),4.05(t,J＝9.2Hz,3H),4.01–3.93(m,1H),3.92–3.84(m,3H),3.82(d,J＝2.5Hz,3H),3.78–3.69(m,4H),3.68–3.59(m,2H),3.58–3.50(m,4H),3.47(dd,J＝9.2,3.6Hz,1H),3.42–3.37(m,1H),3.29–3.16(m,2H),3.13–3.03(m,1H),1.60–1.42(m,4H),1.30(s,2H). 13 C NMR(100MHz,d 6 -Acetone)δ163.33,163.25,163.19,162.91,162.84,162.40,162.32,140.58,140.27,139.80,139.62,139.53,139.50,139.36,138.85,136.46,129.60,129.40,129.29,129.15,129.12,129.04,129.00,128.96,128.95,128.91,128.89,128.87,128.67,128.64,128.60,128.59,128.49,128.45,128.43,128.33,128.31,128.25,128.21,128.16,128.11,128.03,127.98,127.92,105.32,102.38,102.13,102.04,97.36,94.23,94.19,93.85,93.81,93.72,93.68,93.42,93.39,85.40,83.05,79.57,79.34,78.25,77.34,77.10,76.90,76.47,76.19,76.05,75.85,75.61,75.33,75.31,75.27,75.23,75.12,74.94,73.90,73.36,73.21,71.71,71.57,70.58,69.93,69.33,68.14,67.69,67.37,62.23,62.11,58.13,56.39,55.75,50.94,47.99,47.00,30.10,28.82,28.29,24.28.
Compounds 2-4
Compound 2-22 (32 mg,0.0124 mmol) was dissolved in 1mL tetrahydrofuran and 5mL t-butanol, 2mL H were added 2 O, 0.05mL AcOH, 120mg Pd (OH) 2 C (20%) under ice bath H 2 Aeration 15min, H of 1atm 2 Reacting at room temperature for 5 days, filtering, concentrating under reduced pressure, and concentrating with Sephadex LH-20 Column (CH) 3 OH:H 2 O=1:1) to give compound 2-4 (6.7 mg,0.00614mmol, 49%). 1 H NMR(600MHz,D 2 O)δ8.45(s,1H),5.01(d,J＝8.4Hz,1H),4.86(d,J＝3.9Hz,1H),4.61(d,J＝8.5Hz,1H),4.50(d,J＝7.9Hz,1H),4.47(d,J＝8.5Hz,1H),4.36(d,J＝2.4Hz,1H),4.19(d,J＝3.1Hz,1H),4.14–4.08(m,2H),4.02(dd,J＝11.0,10.0Hz,2H),3.95(d,J＝11.9Hz,1H),3.92–3.84(m,4H),3.83–3.73(m,5H),3.73–3.67(m,4H),3.66–3.63(m,1H),3.59(t,J＝9.8Hz,1H),3.54–3.50(m,1H),3.50–3.39(m,6H),3.34–3.30(m,1H),3.02(t,J＝7.5Hz,2H),2.01(dd,J＝7.0,1.5Hz,12H),1.73–1.60(m,4H),1.51–1.41(m,2H). 13 C NMR(150MHz,D 2 O)δ175.55,174.72,174.59,174.57,174.10,104.34,103.63,101.78,101.29,98.00,80.35,80.11,77.38,76.70,75.78,75.69,75.50,74.70,74.10,72.83,71.61,70.48,69.78,69.37,69.31,67.97,67.11,61.22,60.44,55.51,54.64,53.59,51.50,39.38,27.98,26.45,22.48,22.32,22.18,22.15,21.96.
Compound S85
Compound S15 (1.1 g,2 mmol) was dissolved in 10mL DMF, imidazole (272 mg,4mmol,2 eq), dimethyl tert-butylchlorosilane (452 mg,3mmol,1.5 eq) and nitrogen were added in sequence, stirring overnight at room temperature, ethyl acetate dilution and transfer out, washing three times with water, saturated NaHCO 3 The solution and saturated saline were washed once each, anhydrous Na 2 SO 4 Drying, filtration, concentration under reduced pressure, column chromatography (petroleum ether: ethyl acetate 12:1-6:1) gave colorless syrup S84 (1.16 g,1.74mmol,87%, beta configuration). Compound S84 (250 mg,0.37 mmol) was dissolved in dichloromethane: to methanol (2:1, 3 mL), sodium methoxide (10 mg,0.18mmol,0.5 eq) was added, and the mixture was reacted at room temperature for 5 hours, and the pH was adjusted to about 7 by adding an acidic resin (monitored by a pH test paper), filtered, concentrated under reduced pressure, and separated by column chromatography (petroleum ether: ethyl acetate 10:1-5:1) to give colorless syrup S85 (208 mg,0.33mmol, 90%).
Compound S86
Donor S7 (84 mg,0.1mmol,1.5 eq) and acceptor S85 (42 mg,0.067mmol,1 eq) were mixed, toluene azeotroped twice, the water was dried by spinning, and dissolved in 2mL DCMMS drying Water removal x 2), adding +.>MS (200 mg), nitrogen protection, stirring at-40℃for 10min, adding TBSOTf (3. Mu.L, 0.013mmol,0.2 eq), reacting at-40℃for 2 h, adding triethylamine for quenching, suction filtration, concentrating under reduced pressure, and flash column chromatography (petroleum ether: ethyl acetate 12:1-6:1) to give colorless and transparent syrup S86 (73 mg,0.057mmol, 85%).
Compounds 2-20
Compound S86 (950 mg,0.75 mmol) was dissolved in 25mL DCM, 5mL Buffer solution was added, DDQ (0.41 g,1.8mmol,2.4 eq) was added under ice-water bath, gradually warmed to room temperature for 5h, quenched with saturated sodium thiosulfate solution, diluted with ethyl acetate and transferred out, filtered with suction, and the filtrate was washed with water, saturated NaHCO in sequence 3 Solution washing, saturated saline washing and anhydrous Na 2 SO 4 Drying, filtration, concentration under reduced pressure, column chromatography (petroleum ether: ethyl acetate 8:1-2:1) gave colorless syrup 2-20 (596 mg,0.58mmol, 78%).
Compounds 2-49
The acceptor 2-20 (872 mg,0.847 mmol) and the donor 2-7 (973 mg,1.27mmol,1.5 eq) were weighed into a reaction flask, azeotroped 3 times with toluene, and an oil pumpPumping for 2h, adding activatedMS, nitrogen purging 3 times, charging 44mL of dried DCM into the system, stirring at-60deg.C for 10min, charging TfOH (7.5 uL,0.0847mmol,0.1 eq), detecting complete reaction of receptor by TCL after 3.5h, adding Et 3 N-quenching, filtration, concentration, dry-mixing, column chromatography (petroleum ether: ethyl acetate=10:1→3:1) gave foam solid 2-49 (1.27 g,0.79mmol, 93%).
Compounds 2-69
Acceptor 2-49 (110 mg,0.068 mmol) and donor 2-21 (166 mg,0.137mmol,2 eq) were weighed into a reaction flask, azeotroped 3 times with toluene, pumped with oil for 2h, and activated was addedMS, nitrogen purging 3 times, charging 3mL of dry DCM into the system, stirring at 0deg.C for 10min, charging TMSOTF (3 uL,0.014mmol,0.2 eq), adding saturated NaHCO after 2.5h 3 The solution was quenched, filtered and concentrated, separated using a gel column (MeOH: dcm=1:1), then dry loaded, and column chromatographed (petroleum ether: ethyl acetate=10:1→3:1) to give a mixture of foam solids 2-69 and 2-69a (152 mg,0.0578mmol,85%, β/α=4.8:1). Beta configuration products 2-69: [ alpha ] ]D 20 ＝+1.8(c 3.0，CHCl 3 )， 1 H NMR(400MHz,CDCl 3 )δ8.25(d,J＝6.6Hz,1H),7.88(d,J＝8.1Hz,1H),7.35–7.30(m,6H),7.30–7.23(m,27H),7.22–7.17(m,10H),7.16(s,5H),7.12–7.06(m,6H),6.91(d,J＝8.2Hz,1H),5.30(d,J＝8.2Hz,1H),5.19–5.11(m,2H),5.06(d,J＝12.2Hz,1H),5.03–4.98(m,1H),4.81(s,1H),4.78(s,2H),4.76(d,J＝3.0Hz,2H),4.74–4.71(m,2H),4.71–4.66(m,4H),4.66–4.60(m,2H),4.58(d,J＝4.5Hz,1H),4.55(s,1H),4.50(dd,J＝14.3,2.0Hz,2H),4.46(d,J＝5.6Hz,1H),4.44–4.40(m,2H),4.38(s,2H),4.36–4.31(m,1H),4.19(dd,J＝11.9,4.7Hz,1H),4.10–4.02(m,1H),4.00–3.89(m,6H),3.86–3.76(m,6H),3.74–3.66(m,1H),3.62–3.42(m,10H),2.99–2.87(m,1H),2.59–2.48(m,3H),2.41(t,J＝6.3Hz,2H),2.38–2.28(m,1H),2.13–2.05(m,4H),2.03(s,3H),0.77(s,9H),0.07(s,3H),0.00(s,3H). 13 C NMR(100MHz,CDCl 3 )δ208.10,206.47,172.20,171.48,167.85,163.88,162.72,162.44,161.80,139.48,138.49,138.15,137.98,137.95,137.87,137.85,137.48,137.33,134.98,129.13,128.77,128.63,128.55,128.51,128.47,128.46,128.43,128.37,128.24,128.20,128.17,128.08,127.96,127.92,127.84,127.78,127.75,127.72,127.67,127.60,127.55,127.09,126.97,101.85,101.47,99.40,98.67,98.21,92.65,92.47,92.07,91.54,82.38,82.29,80.27,80.13,78.13,77.82,77.62,77.28,76.20,75.29,75.23,75.18,75.14,75.10,74.88,74.79,74.67,73.62,73.32,73.25,73.17,72.96,70.29,68.95,68.46,67.65,62.69,57.69,57.35,57.07,54.81,37.81,37.74,30.05,29.78,27.83,25.83,17.98,-3.66,-4.50.
Compounds 2-23
Compounds 2 to 69 (284 mg,0.202 mmol) were dissolved in 10mL tetrahydrofuran, 1M TBAF tetrahydrofuran solution (0.61 mL,0.606mmol,3 eq), acOH (35 uL,0.606mmol,3 eq) were added, reacted in an oil bath at 30℃and concentrated after 36h, dried and stirred, and column chromatographed (Petroleum ether: ethyl acetate=4:1→1:1) to give 450mg of intermediate; the intermediate was dissolved in 9mL of acetone and Cs was added under ice bath 2 CO 3 (105 mg,0.322mmol,1.6 eq), PTFAI (41 uL,0.27mmol,1.3 eq), gradually warmed to RT and reacted after 3h with Et 3 N, filtration, concentration, dry-mixing, column chromatography (petroleum ether: ethyl acetate=10:1→2.5:1) gave foam solid 2-23 (457 mg,0.168mmol, 84%). 1 H NMR(400MHz,d 6 -Acetone)δ8.49–8.44(m,2H),8.41(d,J＝9.4Hz,1H),8.10(d,J＝9.3Hz,1H),7.44(d,J＝7.2Hz,3H),7.38–7.35(m,4H),7.34–7.32(m,7H),7.32–7.31(m,5H),7.30–7.27(m,10H),7.26(s,8H),7.25–7.22(m,9H),7.21–7.16(m,5H),7.13(t,J＝7.4Hz,1H),7.02(t,J＝7.4Hz,1H),6.57(d,J＝7.7Hz,2H),6.38(s,1H),5.53(d,J＝8.4Hz,1H),5.27(d,J＝12.5Hz,1H),5.20–5.14(m,2H),5.06(dd,J＝9.5,8.1Hz,1H),4.97(d,J＝11.4Hz,1H),4.88–4.83(m,3H),4.83–4.80(m,2H),4.79(s,1H),4.78–4.76(m,3H),4.67(t,J＝4.7Hz,3H),4.63(d,J＝4.5Hz,2H),4.58(d,J＝8.4Hz,3H),4.56–4.49(m,2H),4.47–4.43(m,1H),4.42(d,J＝3.1Hz,1H),4.38(s,1H),4.35(d,J＝1.6Hz,1H),4.33(s,1H),4.32–4.28(m,2H),4.27–4.19(m,2H),4.10(d,J＝9.2Hz,2H),4.07–4.02(m,2H),4.02–3.98(m,2H),3.97–3.93(m,1H),3.90–3.86(m,1H),3.85–3.81(m,3H),3.80–3.72(m,3H),3.68(d,J＝5.1Hz,1H),3.67–3.63(m,2H),3.59–3.53(m,1H),2.85–2.75(m,3H),2.71–2.65(m,2H),2.56–2.45(m,3H),2.15(s,3H),2.05(s,3H). 13 C NMR(100MHz,d 6 Acetone) delta 206.98,206.37,172.01,171.85,168.36,162.51,162.43,162.04,162.00,161.97,161.92,161.75,161.67,143.82,139.15,138.80,138.68,138.66,138.62,138.58,138.41,138.38,138.03,135.63,128.80,128.52,128.50,128.31,128.30,128.27,128.24,128.20,128.18,128.12,128.10,128.04,128.02,127.83,127.75,127.68,127.67,127.65,127.50,127.39,127.35,127.29,127.18,126.91,123.82,119.34,101.69,101.41,101.24,100.70,94.04,93.26,93.22,92.99,92.95,92.85,92.81,92.45,92.41,82.29,81.94,81.91,79.44,78.54,78.11,77.36,77.06,76.60,75.71,75.33,75.18,74.85,74.67,74.56,74.47,74.42,74.26,74.05,73.43,73.38,73.24,73.01,72.48,72.34,69.38,69.08,67.07,62.65,57.17,57.08,56.95,56.86,55.36,55.27,55.00,54.89,54.59,37.67,37.39,29.10,28.89,27.90,27.65 Compounds 2-24
Acceptor 2-22 (113 mg,0.043mmol,1 eq) and donor 2-23 (150 mg,0.0558mmol,1.3 eq) were weighed into a tube sealer and activated was addedMS, sparged 3 times with nitrogen, and 2mL of dried DCM/Et added under nitrogen 2 O (1:3, volume ratio), TBSOTf (2 uL,0.00859mmol,0.2 eq) at 20deg.CIn oil bath, saturated NaHCO is added after 3.5h 3 The solution was quenched, filtered and concentrated, separated using a gel column (MeOH: dcm=1:1), then dry loaded, and column chromatographed (petroleum ether: ethyl acetate=5:1→2:1) to give a mixture of foam solids 2-24 and 2-24b (183 mg,0.0357mmol,83%, β/α=1:1).
Compounds 2-75
A mixture of compounds 2-24 and 2-24b (120 mg,0.0234 mmol) was dissolved in 2.6mL of dichloromethane, 1.3mL of pyridine, 0.8mL of acetic acid were added sequentially with stirring, and 0.12mL of N was added under ice-bath 2 H 4 ·H 2 O/AcOH (volume ratio 1:1), after 6h at room temperature, dichloromethane was diluted, washed with 1M hydrochloric acid and then saturated NaHCO 3 Washing with solution and NaCl solution, anhydrous Na 2 SO 4 Drying, concentrating, dry-mixing, and column chromatography (petroleum ether: ethyl acetate=5:1→2:1) gave foam-like solid 2-75 (55 mg,0.01 mmol, 48%). [ alpha ]]D 20 ＝+20.6(c 1.1，CHCl 3 )， 1 H NMR(600MHz,d 6 -Acetone)δ8.45–8.35(m,2H),8.18(dd,J＝7.7,3.0Hz,1H),8.10(d,J＝7.9Hz,1H),7.94(d,J＝8.5Hz,1H),7.91(d,J＝9.3Hz,1H),7.45–7.39(m,8H),7.34(dd,J＝11.2,4.5Hz,14H),7.32–7.28(m,36H),7.28–7.26(m,18H),7.25(d,J＝2.5Hz,4H),7.25–7.22(m,14H),7.22–7.19(m,7H),7.18–7.16(m,6H),7.16–7.14(m,2H),7.14–7.10(m,2H),7.09–7.05(m,1H),5.47(d,J＝8.4Hz,2H),5.22(dd,J＝11.4,4.1Hz,3H),5.14(s,4H),5.11(d,J＝8.3Hz,1H),5.07(d,J＝3.2Hz,1H),5.04(d,J＝11.6Hz,1H),5.01(d,J＝11.5Hz,1H),4.91(d,J＝11.4Hz,1H),4.88(d,J＝2.8Hz,1H),4.86(d,J＝5.0Hz,1H),4.85–4.82(m,3H),4.81(s,2H),4.79(d,J＝3.2Hz,2H),4.78(d,J＝3.5Hz,1H),4.76(d,J＝4.9Hz,1H),4.74(d,J＝4.4Hz,1H),4.71(d,J＝8.8Hz,2H),4.68(s,1H),4.66–4.64(m,2H),4.64(d,J＝2.4Hz,2H),4.59(d,J＝12.2Hz,1H),4.55(s,3H),4.54–4.53(m,3H),4.52(s,2H),4.51–4.48(m,3H),4.46(d,J＝11.8Hz,2H),4.43(s,1H),4.41(s,1H),4.40(s,1H),4.38–4.35(m,2H),4.33–4.26(m,4H),4.26–4.17(m,5H),4.14–4.08(m,5H),4.08–4.02(m,4H),4.08–3.99(m,8H),4.01(d,J＝9.9Hz,3H),3.98–3.94(m,5H),3.93–3.87(m,3H),3.86–3.81(m,6H),3.80–3.76(m,4H),3.75–3.69(m,4H),3.65–3.60(m,5H),3.57–3.55(m,2H),3.54–3.49(m,6H),3.48–3.46(m,1H),3.45–3.39(m,3H),3.37–3.34(m,1H),3.29–3.19(m,2H),3.11–3.01(m,1H),1.59–1.50(m,4H),1.36–1.32(m,2H). 13 C NMR(150MHz,d 6 -Acetone)δ209.79,209.75,169.09,169.03,163.34,163.32,163.27,163.24,163.20,163.16,162.92,162.86,162.84,162.79,162.52,162.45,162.35,162.28,157.13,156.53,140.56,140.42,140.24,140.03,139.76,139.63,139.52,139.48,139.45,139.43,139.32,139.26,139.18,138.86,138.81,138.40,138.16,136.47,136.45,129.67,129.41,129.36,129.30,129.27,129.23,129.15,129.11,129.09,129.08,129.03,129.02,128.97,128.93,128.92,128.88,128.86,128.65,128.63,128.61,128.57,128.54,128.51,128.48,128.45,128.40,128.34,128.30,128.24,128.22,128.20,128.17,128.14,128.10,128.04,128.00,127.97,127.94,127.90,127.82,106.00,105.32,102.50,102.26,102.15,101.88,101.67,98.01,97.36,97.20,94.39,94.36,94.21,94.17,93.84,93.80,93.77,93.61,93.58,93.42,93.38,93.26,93.23,85.57,85.53,85.31,85.27,82.97,82.87,82.84,80.16,79.58,79.31,79.08,79.05,78.18,78.14,77.92,77.88,77.78,77.48,77.05,76.95,76.48,76.20,76.13,76.10,75.99,75.95,75.87,75.83,75.76,75.72,75.55,75.48,75.42,75.34,75.28,75.26,75.21,75.19,75.09,74.86,73.90,73.88,73.71,73.44,73.29,73.20,73.09,73.06,72.39,72.28,71.73,71.53,71.28,71.04,70.71,70.41,69.94,69.91,69.67,69.26,68.14,68.00,67.65,67.54,67.35,66.75,62.20,62.07,61.98,61.85,58.45,58.36,58.25,58.19,58.14,58.10,57.97,57.10,56.92,56.80,56.58,56.52,56.49,55.93,55.79,55.69,55.46,55.43,50.97,50.76,48.03,47.07,30.36,28.87,28.26,24.28.
Compounds 2-5
Compound 2-75 (30 mg,0.00608 mmol) was dissolved in 1mL tetrahydrofuran and 5mL t-butyl was added Alcohol, 2mL H 2 O, 0.05mL AcOH, 150mg Pd (OH) 2 C (20%) under ice bath H 2 Aeration 15min, H of 1atm 2 Reacting at room temperature for 5 days under ambient condition, filtering, concentrating under reduced pressure, and purifying with Sephadex LH-20 column (CH 3 OH:H 2 O=1:1) to give compound 2-5 (9 mg,0.00432mmol, 71%). 1 H NMR(600MHz,D 2 O)δ4.94(d,J＝8.4Hz,1H),4.91(d,J＝8.4Hz,1H),4.83(d,J＝3.8Hz,1H),4.77(d,J＝3.9Hz,1H),4.52(d,J＝8.5Hz,2H),4.47(d,J＝7.8Hz,1H),4.43(d,J＝7.8Hz,1H),4.41–4.37(m,2H),4.28(d,J＝9.7Hz,2H),4.11(d,J＝2.5Hz,2H),4.07–4.05(m,2H),4.03(d,J＝1.4Hz,1H),4.02(d,J＝4.0Hz,1H),3.98(d,J＝12.0Hz,1H),3.95(d,J＝6.2Hz,1H),3.93(s,1H),3.91(d,J＝2.1Hz,1H),3.88(s,1H),3.86(s,1H),3.84(s,1H),3.81(s,1H),3.79(s,3H),3.77(d,J＝2.9Hz,2H),3.76(d,J＝3.5Hz,1H),3.75–3.73(m,2H),3.71(d,J＝2.8Hz,1H),3.69(s,2H),3.68–3.66(m,4H),3.64(d,J＝5.1Hz,2H),3.63–3.61(m,4H),3.60(d,J＝4.4Hz,2H),3.57–3.54(m,3H),3.51(s,2H),3.50(s,1H),3.48(s,1H),3.46–3.42(m,2H),3.38(s,2H),3.37(s,4H),3.33(d,J＝7.7Hz,2H),3.25–3.22(m,2H),2.93(t,J＝7.5Hz,2H),1.95–1.90(m,24H),1.64–1.58(m,4H),1.40–1.34(m,2H). 13 C NMR(150MHz,D 2 O)δ175.57,175.55,174.73,174.68,174.62,174.57,174.22,174.10,104.69,104.31,103.84,103.61,101.84,101.79,101.31,101.26,98.01,80.56,80.32,80.15,80.11,77.39,76.72,76.67,75.79,75.67,75.59,75.49,74.80,74.63,74.19,74.11,74.01,72.87,72.80,71.61,71.43,70.50,70.38,69.78,69.53,69.38,69.32,68.90,68.03,67.92,67.32,67.16,67.11,64.69,61.69,61.48,61.22,61.14,60.55,60.44,55.60,55.52,54.65,53.58,51.55,51.42,39.39,28.00,26.46,22.50,22.33,22.25,22.19,22.16,21.97.
In summary, the invention completes the synthesis of the Acinetobacter baumannii ATCC 17961 lipopolysaccharide O-antigen, including tetraose, pentaose and decaose molecules, by selecting proper protecting groups and proper synthesis strategies.
For the synthesis of tetraose 2-3, pentaose 2-4 and decaose 2-5 with high branching structure, the invention adopts the method of firstly completing the connection of the diamido glucuronic acid with higher difficulty and galactose C4-hydroxyl, and then completing the stereoselective construction of the galactose C-3 glycosidic bond with dense branching by flexibly adjusting the volume of the donor and the protecting group. It was found in the experiments that the use of the Lev protecting group containing disaccharide donor 2-21 in reaction with the trisaccharide acceptor gives the desired glycoside product in excellent yields and stereoselectivity. Simultaneously realizes the one-pot synthesis of potential pentasaccharide acceptors and donors. Finally, the glycosylation reaction of [5+5] is performed by using the solvent effect, and the decasaccharide molecule is obtained in an excellent yield.
Claims (10)
1. The tetraose compound is characterized by comprising a compound shown in a formula I and a compound shown in a formula II, and the specific structure is as follows:
R 1 TIPS, bn or H; ac is acetyl, bn is benzyl, TIPS is triisopropylsilyl, bz is benzoyl, TCA is trichloroacetic acid, cbz is benzyloxycarbonyl.
2. The compound of claim 1, wherein the tetraose compound of formula II is 2-13, 2-42 or 2-43, having the formula:
3. a pentasaccharide compound is characterized by comprising a compound shown in a formula III, and the specific structure is as follows:
R 2 is Lev, bn or H; r is R 3 Is Lev, bn or H; r is R 4 Is TBS (Tunnel boring system),Or->Bn is benzyl, lev is levulinyl, ph is phenyl, TIPS is triisopropylsilyl, TBS is tert-butyldimethylsilyl, TCA is trichloroacetic acid, cbz is benzyloxycarbonyl.
4. A compound according to claim 3, wherein the compound of formula III is 2-70, 2-22, 2-69 or 2-23, having the formula:
5. a process for the preparation of a compound as claimed in claim 3 or 4, comprising the steps of:
1) Mixing disaccharide donor, trisaccharide acceptor and toluene for co-evaporation to obtain a mixture, adding an organic solvent I into the mixture for pre-reaction, adding a catalyst I, and reacting at room temperature until the acceptor is completely consumed;
2) After the reaction, quenching, suction filtration, decompression concentration to obtain coarse product, and purifying with gel column chromatography and silica gel column chromatography successively to obtain final product.
6. The method of claim 5, wherein the disaccharide donor is 2-21 and has the structural formula:
the trisaccharide receptor is 2-72 and 2-49, and the structural formula is as follows:
7. the process according to claim 5, wherein the organic solvent I is methylene chloride, toluene or chloroform; the catalyst I is TMSOTF;
the molar ratio of disaccharide donor and trisaccharide acceptor is (1-2): 1, a step of; the molar ratio of the trisaccharide acceptor to the catalyst I is (1.6-1.9): 1, a step of; the volume ratio of the organic solvent I to the catalyst I is (1-1.5): 1.
8. use of a pentasaccharide compound according to claim 3 or 4 for the synthesis of a decasaccharide compound.
9. The method for synthesizing a decasaccharide compound by using the pentasaccharide compound according to claim 3 or 4, wherein the decasaccharide compound comprises a compound shown in a formula IV and a compound shown in a formula V, and has the following structure:
R 2 is Lev, bn or H; r is R 3 Is Lev, bn or H; bn is benzyl, lev is levulinyl, TIPS is triisopropylsilyl, TCA is trichloroacetic acid, cbz is benzyloxycarbonyl;
the synthesis method comprises the following steps:
Reacting pentasaccharide acceptor and pentasaccharide donor at 20-25deg.C for 2-4 hr under the condition of catalyst, quenching, filtering, concentrating, separating and purifying to obtain decasaccharide compound.
10. The method of claim 9, wherein the compound of formula V is 2-24, 2-75, and has the structural formula:
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