CN110041383A - 硫酸软骨素寡糖、其制备方法和应用 - Google Patents
硫酸软骨素寡糖、其制备方法和应用 Download PDFInfo
- Publication number
- CN110041383A CN110041383A CN201810033328.1A CN201810033328A CN110041383A CN 110041383 A CN110041383 A CN 110041383A CN 201810033328 A CN201810033328 A CN 201810033328A CN 110041383 A CN110041383 A CN 110041383A
- Authority
- CN
- China
- Prior art keywords
- arc
- pmb
- compound
- chondroitin sulfate
- mmol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 229920001287 Chondroitin sulfate Polymers 0.000 title claims abstract description 73
- 229940059329 chondroitin sulfate Drugs 0.000 title claims abstract description 73
- -1 Chondroitin sulfate oligosaccharides Chemical class 0.000 title claims abstract description 67
- 229920001542 oligosaccharide Polymers 0.000 title claims abstract description 60
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 238000006243 chemical reaction Methods 0.000 claims abstract description 194
- 238000006206 glycosylation reaction Methods 0.000 claims abstract description 18
- 239000000543 intermediate Substances 0.000 claims abstract description 14
- 150000001540 azides Chemical class 0.000 claims abstract description 11
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 39
- 150000001875 compounds Chemical class 0.000 claims description 37
- YXFVVABEGXRONW-UHFFFAOYSA-N toluene Substances CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 37
- 150000002016 disaccharides Chemical class 0.000 claims description 35
- 238000000034 method Methods 0.000 claims description 16
- 150000002772 monosaccharides Chemical class 0.000 claims description 15
- 108010092801 Midkine Proteins 0.000 claims description 14
- 235000000346 sugar Nutrition 0.000 claims description 14
- 150000004044 tetrasaccharides Chemical class 0.000 claims description 14
- 201000010099 disease Diseases 0.000 claims description 12
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 12
- MSWZFWKMSRAUBD-GASJEMHNSA-N 2-amino-2-deoxy-D-galactopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O MSWZFWKMSRAUBD-GASJEMHNSA-N 0.000 claims description 11
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 claims description 11
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 10
- 229920000669 heparin Polymers 0.000 claims description 10
- 229960002897 heparin Drugs 0.000 claims description 10
- 102000004127 Cytokines Human genes 0.000 claims description 9
- 108090000695 Cytokines Proteins 0.000 claims description 9
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 claims description 9
- 239000002904 solvent Substances 0.000 claims description 9
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 8
- 206010028980 Neoplasm Diseases 0.000 claims description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 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 claims description 7
- 230000002378 acidificating effect Effects 0.000 claims description 7
- IKDUDTNKRLTJSI-UHFFFAOYSA-N hydrazine monohydrate Substances O.NN IKDUDTNKRLTJSI-UHFFFAOYSA-N 0.000 claims description 7
- 150000003839 salts Chemical class 0.000 claims description 7
- JOOXCMJARBKPKM-UHFFFAOYSA-N 4-oxopentanoic acid Chemical compound CC(=O)CCC(O)=O JOOXCMJARBKPKM-UHFFFAOYSA-N 0.000 claims description 6
- 238000010511 deprotection reaction Methods 0.000 claims description 5
- 230000002981 neuropathic effect Effects 0.000 claims description 5
- 230000002829 reductive effect Effects 0.000 claims description 5
- 238000005670 sulfation reaction Methods 0.000 claims description 5
- DHKHKXVYLBGOIT-UHFFFAOYSA-N acetaldehyde Diethyl Acetal Natural products CCOC(C)OCC DHKHKXVYLBGOIT-UHFFFAOYSA-N 0.000 claims description 4
- 125000000738 acetamido group Chemical group [H]C([H])([H])C(=O)N([H])[*] 0.000 claims description 4
- 230000000694 effects Effects 0.000 claims description 4
- 125000006239 protecting group Chemical class 0.000 claims description 4
- 230000019635 sulfation Effects 0.000 claims description 4
- ABJSOROVZZKJGI-OCYUSGCXSA-N (1r,2r,4r)-2-(4-bromophenyl)-n-[(4-chlorophenyl)-(2-fluoropyridin-4-yl)methyl]-4-morpholin-4-ylcyclohexane-1-carboxamide Chemical compound C1=NC(F)=CC(C(NC(=O)[C@H]2[C@@H](C[C@@H](CC2)N2CCOCC2)C=2C=CC(Br)=CC=2)C=2C=CC(Cl)=CC=2)=C1 ABJSOROVZZKJGI-OCYUSGCXSA-N 0.000 claims description 3
- STBLNCCBQMHSRC-BATDWUPUSA-N (2s)-n-[(3s,4s)-5-acetyl-7-cyano-4-methyl-1-[(2-methylnaphthalen-1-yl)methyl]-2-oxo-3,4-dihydro-1,5-benzodiazepin-3-yl]-2-(methylamino)propanamide Chemical compound O=C1[C@@H](NC(=O)[C@H](C)NC)[C@H](C)N(C(C)=O)C2=CC(C#N)=CC=C2N1CC1=C(C)C=CC2=CC=CC=C12 STBLNCCBQMHSRC-BATDWUPUSA-N 0.000 claims description 3
- SMNRFWMNPDABKZ-WVALLCKVSA-N [[(2R,3S,4R,5S)-5-(2,6-dioxo-3H-pyridin-3-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [[[(2R,3S,4S,5R,6R)-4-fluoro-3,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-hydroxyphosphoryl]oxy-hydroxyphosphoryl] hydrogen phosphate Chemical compound OC[C@H]1O[C@H](OP(O)(=O)OP(O)(=O)OP(O)(=O)OP(O)(=O)OC[C@H]2O[C@H]([C@H](O)[C@@H]2O)C2C=CC(=O)NC2=O)[C@H](O)[C@@H](F)[C@@H]1O SMNRFWMNPDABKZ-WVALLCKVSA-N 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
- 229940125878 compound 36 Drugs 0.000 claims description 3
- 239000003814 drug Substances 0.000 claims description 3
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 claims description 3
- 229940040102 levulinic acid Drugs 0.000 claims description 3
- ASGMFNBUXDJWJJ-JLCFBVMHSA-N (1R,3R)-3-[[3-bromo-1-[4-(5-methyl-1,3,4-thiadiazol-2-yl)phenyl]pyrazolo[3,4-d]pyrimidin-6-yl]amino]-N,1-dimethylcyclopentane-1-carboxamide Chemical compound BrC1=NN(C2=NC(=NC=C21)N[C@H]1C[C@@](CC1)(C(=O)NC)C)C1=CC=C(C=C1)C=1SC(=NN=1)C ASGMFNBUXDJWJJ-JLCFBVMHSA-N 0.000 claims description 2
- IUSARDYWEPUTPN-OZBXUNDUSA-N (2r)-n-[(2s,3r)-4-[[(4s)-6-(2,2-dimethylpropyl)spiro[3,4-dihydropyrano[2,3-b]pyridine-2,1'-cyclobutane]-4-yl]amino]-3-hydroxy-1-[3-(1,3-thiazol-2-yl)phenyl]butan-2-yl]-2-methoxypropanamide Chemical compound C([C@H](NC(=O)[C@@H](C)OC)[C@H](O)CN[C@@H]1C2=CC(CC(C)(C)C)=CN=C2OC2(CCC2)C1)C(C=1)=CC=CC=1C1=NC=CS1 IUSARDYWEPUTPN-OZBXUNDUSA-N 0.000 claims description 2
- HUWSZNZAROKDRZ-RRLWZMAJSA-N (3r,4r)-3-azaniumyl-5-[[(2s,3r)-1-[(2s)-2,3-dicarboxypyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl]amino]-5-oxo-4-sulfanylpentane-1-sulfonate Chemical compound OS(=O)(=O)CC[C@@H](N)[C@@H](S)C(=O)N[C@@H]([C@H](C)CC)C(=O)N1CCC(C(O)=O)[C@H]1C(O)=O HUWSZNZAROKDRZ-RRLWZMAJSA-N 0.000 claims description 2
- 208000030507 AIDS Diseases 0.000 claims description 2
- 229940127007 Compound 39 Drugs 0.000 claims description 2
- 125000003277 amino group Chemical group 0.000 claims description 2
- 150000001735 carboxylic acids Chemical class 0.000 claims description 2
- 229940125807 compound 37 Drugs 0.000 claims description 2
- 229940127573 compound 38 Drugs 0.000 claims description 2
- 230000032050 esterification Effects 0.000 claims description 2
- 238000005886 esterification reaction Methods 0.000 claims description 2
- PIDFDZJZLOTZTM-KHVQSSSXSA-N ombitasvir Chemical compound COC(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@H]1C(=O)NC1=CC=C([C@H]2N([C@@H](CC2)C=2C=CC(NC(=O)[C@H]3N(CCC3)C(=O)[C@@H](NC(=O)OC)C(C)C)=CC=2)C=2C=CC(=CC=2)C(C)(C)C)C=C1 PIDFDZJZLOTZTM-KHVQSSSXSA-N 0.000 claims description 2
- 150000003138 primary alcohols Chemical class 0.000 claims description 2
- 239000000370 acceptor Substances 0.000 claims 5
- OHBQPCCCRFSCAX-UHFFFAOYSA-N 1,4-Dimethoxybenzene Chemical compound COC1=CC=C(OC)C=C1 OHBQPCCCRFSCAX-UHFFFAOYSA-N 0.000 claims 2
- 102000016776 Midkine Human genes 0.000 claims 2
- 125000004172 4-methoxyphenyl group Chemical group [H]C1=C([H])C(OC([H])([H])[H])=C([H])C([H])=C1* 0.000 claims 1
- JOOXCMJARBKPKM-UHFFFAOYSA-M 4-oxopentanoate Chemical compound CC(=O)CCC([O-])=O JOOXCMJARBKPKM-UHFFFAOYSA-M 0.000 claims 1
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 claims 1
- 239000003638 chemical reducing agent Substances 0.000 claims 1
- 238000006264 debenzylation reaction Methods 0.000 claims 1
- 125000004185 ester group Chemical group 0.000 claims 1
- 229940058352 levulinate Drugs 0.000 claims 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims 1
- 230000011987 methylation Effects 0.000 claims 1
- 238000007069 methylation reaction Methods 0.000 claims 1
- 239000007800 oxidant agent Substances 0.000 claims 1
- 230000001590 oxidative effect Effects 0.000 claims 1
- 238000006699 reductive acetylation reaction Methods 0.000 claims 1
- 230000001180 sulfating effect Effects 0.000 claims 1
- 150000003568 thioethers Chemical class 0.000 claims 1
- 238000003786 synthesis reaction Methods 0.000 abstract description 19
- 230000015572 biosynthetic process Effects 0.000 abstract description 17
- 150000002482 oligosaccharides Chemical class 0.000 abstract description 16
- 230000013595 glycosylation Effects 0.000 abstract description 7
- 230000000144 pharmacologic effect Effects 0.000 abstract description 7
- KOUKXHPPRFNWPP-UHFFFAOYSA-N pyrazine-2,5-dicarboxylic acid;hydrate Chemical compound O.OC(=O)C1=CN=C(C(O)=O)C=N1 KOUKXHPPRFNWPP-UHFFFAOYSA-N 0.000 abstract description 7
- 230000003647 oxidation Effects 0.000 abstract description 6
- 238000001212 derivatisation Methods 0.000 abstract description 4
- 230000009467 reduction Effects 0.000 abstract description 4
- 238000001308 synthesis method Methods 0.000 abstract description 4
- 230000004071 biological effect Effects 0.000 abstract description 3
- ZJLMKPKYJBQJNH-UHFFFAOYSA-N propane-1,3-dithiol Chemical compound SCCCS ZJLMKPKYJBQJNH-UHFFFAOYSA-N 0.000 abstract description 3
- 238000005556 structure-activity relationship Methods 0.000 abstract description 3
- 230000007246 mechanism Effects 0.000 abstract description 2
- 150000001720 carbohydrates Chemical class 0.000 abstract 1
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 267
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 141
- 239000000243 solution Substances 0.000 description 123
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 120
- 238000005481 NMR spectroscopy Methods 0.000 description 79
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 60
- 238000004440 column chromatography Methods 0.000 description 59
- 238000003756 stirring Methods 0.000 description 58
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 45
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 42
- 239000007787 solid Substances 0.000 description 38
- 239000011734 sodium Substances 0.000 description 37
- 239000012074 organic phase Substances 0.000 description 32
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 29
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 description 27
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 27
- 238000012544 monitoring process Methods 0.000 description 27
- 239000003208 petroleum Substances 0.000 description 26
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 21
- 125000001160 methoxycarbonyl group Chemical group [H]C([H])([H])OC(*)=O 0.000 description 21
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 21
- 125000000649 benzylidene group Chemical group [H]C(=[*])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 19
- 239000002808 molecular sieve Substances 0.000 description 18
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 18
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 17
- 238000010791 quenching Methods 0.000 description 17
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 16
- 239000000706 filtrate Substances 0.000 description 16
- 239000006260 foam Substances 0.000 description 16
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 15
- 238000012805 post-processing Methods 0.000 description 15
- 239000011347 resin Substances 0.000 description 15
- 229920005989 resin Polymers 0.000 description 15
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 14
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical class O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 14
- DTQVDTLACAAQTR-UHFFFAOYSA-N trifluoroacetic acid Substances OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 14
- 102100030335 Midkine Human genes 0.000 description 12
- 238000004949 mass spectrometry Methods 0.000 description 12
- 239000002243 precursor Substances 0.000 description 12
- 239000006188 syrup Substances 0.000 description 12
- 235000020357 syrup Nutrition 0.000 description 12
- 238000001914 filtration Methods 0.000 description 11
- 229920006395 saturated elastomer Polymers 0.000 description 11
- 239000012043 crude product Substances 0.000 description 10
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 10
- 229910052739 hydrogen Inorganic materials 0.000 description 9
- 239000000203 mixture Substances 0.000 description 9
- 239000002994 raw material Substances 0.000 description 9
- FMKGJQHNYMWDFJ-CVEARBPZSA-N 2-[[4-(2,2-difluoropropoxy)pyrimidin-5-yl]methylamino]-4-[[(1R,4S)-4-hydroxy-3,3-dimethylcyclohexyl]amino]pyrimidine-5-carbonitrile Chemical compound FC(COC1=NC=NC=C1CNC1=NC=C(C(=N1)N[C@H]1CC([C@H](CC1)O)(C)C)C#N)(C)F FMKGJQHNYMWDFJ-CVEARBPZSA-N 0.000 description 8
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 8
- WQDUMFSSJAZKTM-UHFFFAOYSA-N Sodium methoxide Chemical compound [Na+].[O-]C WQDUMFSSJAZKTM-UHFFFAOYSA-N 0.000 description 8
- WTEOIRVLGSZEPR-UHFFFAOYSA-N boron trifluoride Chemical compound FB(F)F WTEOIRVLGSZEPR-UHFFFAOYSA-N 0.000 description 8
- 229940127113 compound 57 Drugs 0.000 description 8
- 229960001031 glucose Drugs 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- 238000011160 research Methods 0.000 description 8
- 239000000741 silica gel Substances 0.000 description 8
- 229910002027 silica gel Inorganic materials 0.000 description 8
- 229910052786 argon Inorganic materials 0.000 description 7
- 229920001429 chelating resin Polymers 0.000 description 7
- 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 7
- 230000007935 neutral effect Effects 0.000 description 7
- 238000000746 purification Methods 0.000 description 7
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 7
- 235000017557 sodium bicarbonate Nutrition 0.000 description 7
- FTVLMFQEYACZNP-UHFFFAOYSA-N trimethylsilyl trifluoromethanesulfonate Chemical compound C[Si](C)(C)OS(=O)(=O)C(F)(F)F FTVLMFQEYACZNP-UHFFFAOYSA-N 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 229920005654 Sephadex Polymers 0.000 description 6
- 239000012507 Sephadex™ Substances 0.000 description 6
- 239000008346 aqueous phase Substances 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 239000011259 mixed solution Substances 0.000 description 6
- KJIFKLIQANRMOU-UHFFFAOYSA-N oxidanium;4-methylbenzenesulfonate Chemical compound O.CC1=CC=C(S(O)(=O)=O)C=C1 KJIFKLIQANRMOU-UHFFFAOYSA-N 0.000 description 6
- 101100391174 Dictyostelium discoideum forC gene Proteins 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
- 102000001708 Protein Isoforms Human genes 0.000 description 5
- 108010029485 Protein Isoforms Proteins 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 238000010168 coupling process Methods 0.000 description 5
- 150000004676 glycans Chemical class 0.000 description 5
- 230000003993 interaction Effects 0.000 description 5
- 229920001282 polysaccharide Polymers 0.000 description 5
- 239000005017 polysaccharide Substances 0.000 description 5
- 229910000027 potassium carbonate Inorganic materials 0.000 description 5
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 5
- 238000006257 total synthesis reaction Methods 0.000 description 5
- KQZLRWGGWXJPOS-NLFPWZOASA-N 1-[(1R)-1-(2,4-dichlorophenyl)ethyl]-6-[(4S,5R)-4-[(2S)-2-(hydroxymethyl)pyrrolidin-1-yl]-5-methylcyclohexen-1-yl]pyrazolo[3,4-b]pyrazine-3-carbonitrile Chemical compound ClC1=C(C=CC(=C1)Cl)[C@@H](C)N1N=C(C=2C1=NC(=CN=2)C1=CC[C@@H]([C@@H](C1)C)N1[C@@H](CCC1)CO)C#N KQZLRWGGWXJPOS-NLFPWZOASA-N 0.000 description 4
- TVTJUIAKQFIXCE-HUKYDQBMSA-N 2-amino-9-[(2R,3S,4S,5R)-4-fluoro-3-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-7-prop-2-ynyl-1H-purine-6,8-dione Chemical compound NC=1NC(C=2N(C(N(C=2N=1)[C@@H]1O[C@@H]([C@H]([C@H]1O)F)CO)=O)CC#C)=O TVTJUIAKQFIXCE-HUKYDQBMSA-N 0.000 description 4
- NPRYCHLHHVWLQZ-TURQNECASA-N 2-amino-9-[(2R,3S,4S,5R)-4-fluoro-3-hydroxy-5-(hydroxymethyl)oxolan-2-yl]-7-prop-2-ynylpurin-8-one Chemical compound NC1=NC=C2N(C(N(C2=N1)[C@@H]1O[C@@H]([C@H]([C@H]1O)F)CO)=O)CC#C NPRYCHLHHVWLQZ-TURQNECASA-N 0.000 description 4
- 229910015900 BF3 Inorganic materials 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 229940125851 compound 27 Drugs 0.000 description 4
- 229940125877 compound 31 Drugs 0.000 description 4
- 238000005859 coupling reaction Methods 0.000 description 4
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000005342 ion exchange Methods 0.000 description 4
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 235000019345 sodium thiosulphate Nutrition 0.000 description 4
- WJKHJLXJJJATHN-UHFFFAOYSA-N triflic anhydride Chemical compound FC(F)(F)S(=O)(=O)OS(=O)(=O)C(F)(F)F WJKHJLXJJJATHN-UHFFFAOYSA-N 0.000 description 4
- OOKAZRDERJMRCJ-KOUAFAAESA-N (3r)-7-[(1s,2s,4ar,6s,8s)-2,6-dimethyl-8-[(2s)-2-methylbutanoyl]oxy-1,2,4a,5,6,7,8,8a-octahydronaphthalen-1-yl]-3-hydroxy-5-oxoheptanoic acid Chemical compound C1=C[C@H](C)[C@H](CCC(=O)C[C@@H](O)CC(O)=O)C2[C@@H](OC(=O)[C@@H](C)CC)C[C@@H](C)C[C@@H]21 OOKAZRDERJMRCJ-KOUAFAAESA-N 0.000 description 3
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 3
- NNWFSRBWMOZDAK-UHFFFAOYSA-N 2-[2-(carboxymethyl)-3-iodophenyl]acetic acid Chemical compound OC(=O)CC1=CC=CC(I)=C1CC(O)=O NNWFSRBWMOZDAK-UHFFFAOYSA-N 0.000 description 3
- LFOIDLOIBZFWDO-UHFFFAOYSA-N 2-methoxy-6-[6-methoxy-4-[(3-phenylmethoxyphenyl)methoxy]-1-benzofuran-2-yl]imidazo[2,1-b][1,3,4]thiadiazole Chemical compound N1=C2SC(OC)=NN2C=C1C(OC1=CC(OC)=C2)=CC1=C2OCC(C=1)=CC=CC=1OCC1=CC=CC=C1 LFOIDLOIBZFWDO-UHFFFAOYSA-N 0.000 description 3
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 3
- 229940126639 Compound 33 Drugs 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 108090000368 Fibroblast growth factor 8 Proteins 0.000 description 3
- 102100039856 Histone H1.1 Human genes 0.000 description 3
- 102100039855 Histone H1.2 Human genes 0.000 description 3
- 101001035402 Homo sapiens Histone H1.1 Proteins 0.000 description 3
- 101001035375 Homo sapiens Histone H1.2 Proteins 0.000 description 3
- PNUZDKCDAWUEGK-CYZMBNFOSA-N Sitafloxacin Chemical compound C([C@H]1N)N(C=2C(=C3C(C(C(C(O)=O)=CN3[C@H]3[C@H](C3)F)=O)=CC=2F)Cl)CC11CC1 PNUZDKCDAWUEGK-CYZMBNFOSA-N 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 230000008878 coupling Effects 0.000 description 3
- 238000005858 glycosidation reaction Methods 0.000 description 3
- 239000005457 ice water Substances 0.000 description 3
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 description 3
- 239000003456 ion exchange resin Substances 0.000 description 3
- 229920003303 ion-exchange polymer Polymers 0.000 description 3
- ZBELDPMWYXDLNY-UHFFFAOYSA-N methyl 9-(4-bromo-2-fluoroanilino)-[1,3]thiazolo[5,4-f]quinazoline-2-carboximidate Chemical compound C12=C3SC(C(=N)OC)=NC3=CC=C2N=CN=C1NC1=CC=C(Br)C=C1F ZBELDPMWYXDLNY-UHFFFAOYSA-N 0.000 description 3
- NWVVVBRKAWDGAB-UHFFFAOYSA-N p-methoxyphenol Chemical compound COC1=CC=C(O)C=C1 NWVVVBRKAWDGAB-UHFFFAOYSA-N 0.000 description 3
- 102000005162 pleiotrophin Human genes 0.000 description 3
- 238000001953 recrystallisation Methods 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 150000004043 trisaccharides Chemical class 0.000 description 3
- UAOUIVVJBYDFKD-XKCDOFEDSA-N (1R,9R,10S,11R,12R,15S,18S,21R)-10,11,21-trihydroxy-8,8-dimethyl-14-methylidene-4-(prop-2-enylamino)-20-oxa-5-thia-3-azahexacyclo[9.7.2.112,15.01,9.02,6.012,18]henicosa-2(6),3-dien-13-one Chemical compound C([C@@H]1[C@@H](O)[C@@]23C(C1=C)=O)C[C@H]2[C@]12C(N=C(NCC=C)S4)=C4CC(C)(C)[C@H]1[C@H](O)[C@]3(O)OC2 UAOUIVVJBYDFKD-XKCDOFEDSA-N 0.000 description 2
- CBOJBBMQJBVCMW-NQZVPSPJSA-N (2r,3r,4r,5r)-2-amino-3,4,5,6-tetrahydroxyhexanal;hydrochloride Chemical compound Cl.O=C[C@H](N)[C@@H](O)[C@@H](O)[C@H](O)CO CBOJBBMQJBVCMW-NQZVPSPJSA-N 0.000 description 2
- FNHHVPPSBFQMEL-KQHDFZBMSA-N (3S)-5-N-[(1S,5R)-3-hydroxy-6-bicyclo[3.1.0]hexanyl]-7-N,3-dimethyl-3-phenyl-2H-1-benzofuran-5,7-dicarboxamide Chemical compound CNC(=O)c1cc(cc2c1OC[C@@]2(C)c1ccccc1)C(=O)NC1[C@H]2CC(O)C[C@@H]12 FNHHVPPSBFQMEL-KQHDFZBMSA-N 0.000 description 2
- UDQTXCHQKHIQMH-KYGLGHNPSA-N (3ar,5s,6s,7r,7ar)-5-(difluoromethyl)-2-(ethylamino)-5,6,7,7a-tetrahydro-3ah-pyrano[3,2-d][1,3]thiazole-6,7-diol Chemical compound S1C(NCC)=N[C@H]2[C@@H]1O[C@H](C(F)F)[C@@H](O)[C@@H]2O UDQTXCHQKHIQMH-KYGLGHNPSA-N 0.000 description 2
- MPDDTAJMJCESGV-CTUHWIOQSA-M (3r,5r)-7-[2-(4-fluorophenyl)-5-[methyl-[(1r)-1-phenylethyl]carbamoyl]-4-propan-2-ylpyrazol-3-yl]-3,5-dihydroxyheptanoate Chemical compound C1([C@@H](C)N(C)C(=O)C2=NN(C(CC[C@@H](O)C[C@@H](O)CC([O-])=O)=C2C(C)C)C=2C=CC(F)=CC=2)=CC=CC=C1 MPDDTAJMJCESGV-CTUHWIOQSA-M 0.000 description 2
- YQOLEILXOBUDMU-KRWDZBQOSA-N (4R)-5-[(6-bromo-3-methyl-2-pyrrolidin-1-ylquinoline-4-carbonyl)amino]-4-(2-chlorophenyl)pentanoic acid Chemical compound CC1=C(C2=C(C=CC(=C2)Br)N=C1N3CCCC3)C(=O)NC[C@H](CCC(=O)O)C4=CC=CC=C4Cl YQOLEILXOBUDMU-KRWDZBQOSA-N 0.000 description 2
- STPKWKPURVSAJF-LJEWAXOPSA-N (4r,5r)-5-[4-[[4-(1-aza-4-azoniabicyclo[2.2.2]octan-4-ylmethyl)phenyl]methoxy]phenyl]-3,3-dibutyl-7-(dimethylamino)-1,1-dioxo-4,5-dihydro-2h-1$l^{6}-benzothiepin-4-ol Chemical compound O[C@H]1C(CCCC)(CCCC)CS(=O)(=O)C2=CC=C(N(C)C)C=C2[C@H]1C(C=C1)=CC=C1OCC(C=C1)=CC=C1C[N+]1(CC2)CCN2CC1 STPKWKPURVSAJF-LJEWAXOPSA-N 0.000 description 2
- MHSLDASSAFCCDO-UHFFFAOYSA-N 1-(5-tert-butyl-2-methylpyrazol-3-yl)-3-(4-pyridin-4-yloxyphenyl)urea Chemical compound CN1N=C(C(C)(C)C)C=C1NC(=O)NC(C=C1)=CC=C1OC1=CC=NC=C1 MHSLDASSAFCCDO-UHFFFAOYSA-N 0.000 description 2
- WGFNXGPBPIJYLI-UHFFFAOYSA-N 2,6-difluoro-3-[(3-fluorophenyl)sulfonylamino]-n-(3-methoxy-1h-pyrazolo[3,4-b]pyridin-5-yl)benzamide Chemical compound C1=C2C(OC)=NNC2=NC=C1NC(=O)C(C=1F)=C(F)C=CC=1NS(=O)(=O)C1=CC=CC(F)=C1 WGFNXGPBPIJYLI-UHFFFAOYSA-N 0.000 description 2
- FQMZXMVHHKXGTM-UHFFFAOYSA-N 2-(1-adamantyl)-n-[2-[2-(2-hydroxyethylamino)ethylamino]quinolin-5-yl]acetamide Chemical compound C1C(C2)CC(C3)CC2CC13CC(=O)NC1=CC=CC2=NC(NCCNCCO)=CC=C21 FQMZXMVHHKXGTM-UHFFFAOYSA-N 0.000 description 2
- VCUXVXLUOHDHKK-UHFFFAOYSA-N 2-(2-aminopyrimidin-4-yl)-4-(2-chloro-4-methoxyphenyl)-1,3-thiazole-5-carboxamide Chemical compound ClC1=CC(OC)=CC=C1C1=C(C(N)=O)SC(C=2N=C(N)N=CC=2)=N1 VCUXVXLUOHDHKK-UHFFFAOYSA-N 0.000 description 2
- PYRKKGOKRMZEIT-UHFFFAOYSA-N 2-[6-(2-cyclopropylethoxy)-9-(2-hydroxy-2-methylpropyl)-1h-phenanthro[9,10-d]imidazol-2-yl]-5-fluorobenzene-1,3-dicarbonitrile Chemical compound C1=C2C3=CC(CC(C)(O)C)=CC=C3C=3NC(C=4C(=CC(F)=CC=4C#N)C#N)=NC=3C2=CC=C1OCCC1CC1 PYRKKGOKRMZEIT-UHFFFAOYSA-N 0.000 description 2
- VVCMGAUPZIKYTH-VGHSCWAPSA-N 2-acetyloxybenzoic acid;[(2s,3r)-4-(dimethylamino)-3-methyl-1,2-diphenylbutan-2-yl] propanoate;1,3,7-trimethylpurine-2,6-dione Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O.CN1C(=O)N(C)C(=O)C2=C1N=CN2C.C([C@](OC(=O)CC)([C@H](C)CN(C)C)C=1C=CC=CC=1)C1=CC=CC=C1 VVCMGAUPZIKYTH-VGHSCWAPSA-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
- WYFCZWSWFGJODV-MIANJLSGSA-N 4-[[(1s)-2-[(e)-3-[3-chloro-2-fluoro-6-(tetrazol-1-yl)phenyl]prop-2-enoyl]-5-(4-methyl-2-oxopiperazin-1-yl)-3,4-dihydro-1h-isoquinoline-1-carbonyl]amino]benzoic acid Chemical compound O=C1CN(C)CCN1C1=CC=CC2=C1CCN(C(=O)\C=C\C=1C(=CC=C(Cl)C=1F)N1N=NN=C1)[C@@H]2C(=O)NC1=CC=C(C(O)=O)C=C1 WYFCZWSWFGJODV-MIANJLSGSA-N 0.000 description 2
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 2
- WLHCBQAPPJAULW-UHFFFAOYSA-N 4-methylbenzenethiol Chemical compound CC1=CC=C(S)C=C1 WLHCBQAPPJAULW-UHFFFAOYSA-N 0.000 description 2
- XFJBGINZIMNZBW-CRAIPNDOSA-N 5-chloro-2-[4-[(1r,2s)-2-[2-(5-methylsulfonylpyridin-2-yl)oxyethyl]cyclopropyl]piperidin-1-yl]pyrimidine Chemical compound N1=CC(S(=O)(=O)C)=CC=C1OCC[C@H]1[C@@H](C2CCN(CC2)C=2N=CC(Cl)=CN=2)C1 XFJBGINZIMNZBW-CRAIPNDOSA-N 0.000 description 2
- GDUANFXPOZTYKS-UHFFFAOYSA-N 6-bromo-8-[(2,6-difluoro-4-methoxybenzoyl)amino]-4-oxochromene-2-carboxylic acid Chemical compound FC1=CC(OC)=CC(F)=C1C(=O)NC1=CC(Br)=CC2=C1OC(C(O)=O)=CC2=O GDUANFXPOZTYKS-UHFFFAOYSA-N 0.000 description 2
- JQUCWIWWWKZNCS-LESHARBVSA-N C(C1=CC=CC=C1)(=O)NC=1SC[C@H]2[C@@](N1)(CO[C@H](C2)C)C=2SC=C(N2)NC(=O)C2=NC=C(C=C2)OC(F)F Chemical compound C(C1=CC=CC=C1)(=O)NC=1SC[C@H]2[C@@](N1)(CO[C@H](C2)C)C=2SC=C(N2)NC(=O)C2=NC=C(C=C2)OC(F)F JQUCWIWWWKZNCS-LESHARBVSA-N 0.000 description 2
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- 102100027368 Histone H1.3 Human genes 0.000 description 2
- 101001009450 Homo sapiens Histone H1.3 Proteins 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- LVDRREOUMKACNJ-BKMJKUGQSA-N N-[(2R,3S)-2-(4-chlorophenyl)-1-(1,4-dimethyl-2-oxoquinolin-7-yl)-6-oxopiperidin-3-yl]-2-methylpropane-1-sulfonamide Chemical compound CC(C)CS(=O)(=O)N[C@H]1CCC(=O)N([C@@H]1c1ccc(Cl)cc1)c1ccc2c(C)cc(=O)n(C)c2c1 LVDRREOUMKACNJ-BKMJKUGQSA-N 0.000 description 2
- QOVYHDHLFPKQQG-NDEPHWFRSA-N N[C@@H](CCC(=O)N1CCC(CC1)NC1=C2C=CC=CC2=NC(NCC2=CN(CCCNCCCNC3CCCCC3)N=N2)=N1)C(O)=O Chemical compound N[C@@H](CCC(=O)N1CCC(CC1)NC1=C2C=CC=CC2=NC(NCC2=CN(CCCNCCCNC3CCCCC3)N=N2)=N1)C(O)=O QOVYHDHLFPKQQG-NDEPHWFRSA-N 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 2
- DRUIESSIVFYOMK-UHFFFAOYSA-N Trichloroacetonitrile Chemical compound ClC(Cl)(Cl)C#N DRUIESSIVFYOMK-UHFFFAOYSA-N 0.000 description 2
- LJOOWESTVASNOG-UFJKPHDISA-N [(1s,3r,4ar,7s,8s,8as)-3-hydroxy-8-[2-[(4r)-4-hydroxy-6-oxooxan-2-yl]ethyl]-7-methyl-1,2,3,4,4a,7,8,8a-octahydronaphthalen-1-yl] (2s)-2-methylbutanoate Chemical compound C([C@H]1[C@@H](C)C=C[C@H]2C[C@@H](O)C[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)CC1C[C@@H](O)CC(=O)O1 LJOOWESTVASNOG-UFJKPHDISA-N 0.000 description 2
- SPXSEZMVRJLHQG-XMMPIXPASA-N [(2R)-1-[[4-[(3-phenylmethoxyphenoxy)methyl]phenyl]methyl]pyrrolidin-2-yl]methanol Chemical compound C(C1=CC=CC=C1)OC=1C=C(OCC2=CC=C(CN3[C@H](CCC3)CO)C=C2)C=CC=1 SPXSEZMVRJLHQG-XMMPIXPASA-N 0.000 description 2
- PSLUFJFHTBIXMW-WYEYVKMPSA-N [(3r,4ar,5s,6s,6as,10s,10ar,10bs)-3-ethenyl-10,10b-dihydroxy-3,4a,7,7,10a-pentamethyl-1-oxo-6-(2-pyridin-2-ylethylcarbamoyloxy)-5,6,6a,8,9,10-hexahydro-2h-benzo[f]chromen-5-yl] acetate Chemical compound O([C@@H]1[C@@H]([C@]2(O[C@](C)(CC(=O)[C@]2(O)[C@@]2(C)[C@@H](O)CCC(C)(C)[C@@H]21)C=C)C)OC(=O)C)C(=O)NCCC1=CC=CC=N1 PSLUFJFHTBIXMW-WYEYVKMPSA-N 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- PASDCCFISLVPSO-UHFFFAOYSA-N benzoyl chloride Chemical compound ClC(=O)C1=CC=CC=C1 PASDCCFISLVPSO-UHFFFAOYSA-N 0.000 description 2
- 230000008827 biological function Effects 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 229940127204 compound 29 Drugs 0.000 description 2
- 229940125936 compound 42 Drugs 0.000 description 2
- 229940125844 compound 46 Drugs 0.000 description 2
- 229940127271 compound 49 Drugs 0.000 description 2
- BJXYHBKEQFQVES-NWDGAFQWSA-N enpatoran Chemical compound N[C@H]1CN(C[C@H](C1)C(F)(F)F)C1=C2C=CC=NC2=C(C=C1)C#N BJXYHBKEQFQVES-NWDGAFQWSA-N 0.000 description 2
- GWNFQAKCJYEJEW-UHFFFAOYSA-N ethyl 3-[8-[[4-methyl-5-[(3-methyl-4-oxophthalazin-1-yl)methyl]-1,2,4-triazol-3-yl]sulfanyl]octanoylamino]benzoate Chemical compound CCOC(=O)C1=CC(NC(=O)CCCCCCCSC2=NN=C(CC3=NN(C)C(=O)C4=CC=CC=C34)N2C)=CC=C1 GWNFQAKCJYEJEW-UHFFFAOYSA-N 0.000 description 2
- 230000020764 fibrinolysis Effects 0.000 description 2
- 229940097043 glucuronic acid Drugs 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- YGBMCLDVRUGXOV-UHFFFAOYSA-N n-[6-[6-chloro-5-[(4-fluorophenyl)sulfonylamino]pyridin-3-yl]-1,3-benzothiazol-2-yl]acetamide Chemical compound C1=C2SC(NC(=O)C)=NC2=CC=C1C(C=1)=CN=C(Cl)C=1NS(=O)(=O)C1=CC=C(F)C=C1 YGBMCLDVRUGXOV-UHFFFAOYSA-N 0.000 description 2
- IOMMMLWIABWRKL-WUTDNEBXSA-N nazartinib Chemical compound C1N(C(=O)/C=C/CN(C)C)CCCC[C@H]1N1C2=C(Cl)C=CC=C2N=C1NC(=O)C1=CC=NC(C)=C1 IOMMMLWIABWRKL-WUTDNEBXSA-N 0.000 description 2
- 230000001766 physiological effect Effects 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 238000010898 silica gel chromatography Methods 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 150000008163 sugars Chemical class 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 230000002194 synthesizing effect Effects 0.000 description 2
- 238000010189 synthetic method Methods 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- WCTAGTRAWPDFQO-UHFFFAOYSA-K trisodium;hydrogen carbonate;carbonate Chemical compound [Na+].[Na+].[Na+].OC([O-])=O.[O-]C([O-])=O WCTAGTRAWPDFQO-UHFFFAOYSA-K 0.000 description 2
- 238000010626 work up procedure Methods 0.000 description 2
- VIJSPAIQWVPKQZ-BLECARSGSA-N (2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-acetamido-5-(diaminomethylideneamino)pentanoyl]amino]-4-methylpentanoyl]amino]-4,4-dimethylpentanoyl]amino]-4-methylpentanoyl]amino]propanoyl]amino]-5-(diaminomethylideneamino)pentanoic acid Chemical compound NC(=N)NCCC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(C)(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCNC(N)=N)NC(C)=O VIJSPAIQWVPKQZ-BLECARSGSA-N 0.000 description 1
- DEVSOMFAQLZNKR-RJRFIUFISA-N (z)-3-[3-[3,5-bis(trifluoromethyl)phenyl]-1,2,4-triazol-1-yl]-n'-pyrazin-2-ylprop-2-enehydrazide Chemical compound FC(F)(F)C1=CC(C(F)(F)F)=CC(C2=NN(\C=C/C(=O)NNC=3N=CC=NC=3)C=N2)=C1 DEVSOMFAQLZNKR-RJRFIUFISA-N 0.000 description 1
- KKHFRAFPESRGGD-UHFFFAOYSA-N 1,3-dimethyl-7-[3-(n-methylanilino)propyl]purine-2,6-dione Chemical compound C1=NC=2N(C)C(=O)N(C)C(=O)C=2N1CCCN(C)C1=CC=CC=C1 KKHFRAFPESRGGD-UHFFFAOYSA-N 0.000 description 1
- NNHYAHOTXLASEA-UHFFFAOYSA-N 1-(dimethoxymethyl)-4-methoxybenzene Chemical compound COC(OC)C1=CC=C(OC)C=C1 NNHYAHOTXLASEA-UHFFFAOYSA-N 0.000 description 1
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 1
- WZZBNLYBHUDSHF-DHLKQENFSA-N 1-[(3s,4s)-4-[8-(2-chloro-4-pyrimidin-2-yloxyphenyl)-7-fluoro-2-methylimidazo[4,5-c]quinolin-1-yl]-3-fluoropiperidin-1-yl]-2-hydroxyethanone Chemical compound CC1=NC2=CN=C3C=C(F)C(C=4C(=CC(OC=5N=CC=CN=5)=CC=4)Cl)=CC3=C2N1[C@H]1CCN(C(=O)CO)C[C@@H]1F WZZBNLYBHUDSHF-DHLKQENFSA-N 0.000 description 1
- VSTXCZGEEVFJES-UHFFFAOYSA-N 1-cycloundecyl-1,5-diazacycloundec-5-ene Chemical compound C1CCCCCC(CCCC1)N1CCCCCC=NCCC1 VSTXCZGEEVFJES-UHFFFAOYSA-N 0.000 description 1
- SDTORDSXCYSNTD-UHFFFAOYSA-N 1-methoxy-4-[(4-methoxyphenyl)methoxymethyl]benzene Chemical compound C1=CC(OC)=CC=C1COCC1=CC=C(OC)C=C1 SDTORDSXCYSNTD-UHFFFAOYSA-N 0.000 description 1
- QEBYEVQKHRUYPE-UHFFFAOYSA-N 2-(2-chlorophenyl)-5-[(1-methylpyrazol-3-yl)methyl]-4-[[methyl(pyridin-3-ylmethyl)amino]methyl]-1h-pyrazolo[4,3-c]pyridine-3,6-dione Chemical compound C1=CN(C)N=C1CN1C(=O)C=C2NN(C=3C(=CC=CC=3)Cl)C(=O)C2=C1CN(C)CC1=CC=CN=C1 QEBYEVQKHRUYPE-UHFFFAOYSA-N 0.000 description 1
- WFIYPADYPQQLNN-UHFFFAOYSA-N 2-[2-(4-bromopyrazol-1-yl)ethyl]isoindole-1,3-dione Chemical compound C1=C(Br)C=NN1CCN1C(=O)C2=CC=CC=C2C1=O WFIYPADYPQQLNN-UHFFFAOYSA-N 0.000 description 1
- GOJUJUVQIVIZAV-UHFFFAOYSA-N 2-amino-4,6-dichloropyrimidine-5-carbaldehyde Chemical group NC1=NC(Cl)=C(C=O)C(Cl)=N1 GOJUJUVQIVIZAV-UHFFFAOYSA-N 0.000 description 1
- DFRAKBCRUYUFNT-UHFFFAOYSA-N 3,8-dicyclohexyl-2,4,7,9-tetrahydro-[1,3]oxazino[5,6-h][1,3]benzoxazine Chemical compound C1CCCCC1N1CC(C=CC2=C3OCN(C2)C2CCCCC2)=C3OC1 DFRAKBCRUYUFNT-UHFFFAOYSA-N 0.000 description 1
- QCHPKSFMDHPSNR-UHFFFAOYSA-N 3-aminoisobutyric acid Chemical compound NCC(C)C(O)=O QCHPKSFMDHPSNR-UHFFFAOYSA-N 0.000 description 1
- HRADVHZVMOMEPU-UHFFFAOYSA-N 3-iodopyrrolidine-2,5-dione Chemical compound IC1CC(=O)NC1=O HRADVHZVMOMEPU-UHFFFAOYSA-N 0.000 description 1
- VKLKXFOZNHEBSW-UHFFFAOYSA-N 5-[[3-[(4-morpholin-4-ylbenzoyl)amino]phenyl]methoxy]pyridine-3-carboxamide Chemical compound O1CCN(CC1)C1=CC=C(C(=O)NC=2C=C(COC=3C=NC=C(C(=O)N)C=3)C=CC=2)C=C1 VKLKXFOZNHEBSW-UHFFFAOYSA-N 0.000 description 1
- RSIWALKZYXPAGW-NSHDSACASA-N 6-(3-fluorophenyl)-3-methyl-7-[(1s)-1-(7h-purin-6-ylamino)ethyl]-[1,3]thiazolo[3,2-a]pyrimidin-5-one Chemical compound C=1([C@@H](NC=2C=3N=CNC=3N=CN=2)C)N=C2SC=C(C)N2C(=O)C=1C1=CC=CC(F)=C1 RSIWALKZYXPAGW-NSHDSACASA-N 0.000 description 1
- XASOHFCUIQARJT-UHFFFAOYSA-N 8-methoxy-6-[7-(2-morpholin-4-ylethoxy)imidazo[1,2-a]pyridin-3-yl]-2-(2,2,2-trifluoroethyl)-3,4-dihydroisoquinolin-1-one Chemical compound C(N1C(=O)C2=C(OC)C=C(C=3N4C(=NC=3)C=C(C=C4)OCCN3CCOCC3)C=C2CC1)C(F)(F)F XASOHFCUIQARJT-UHFFFAOYSA-N 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 208000020084 Bone disease Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- PKMUHQIDVVOXHQ-HXUWFJFHSA-N C[C@H](C1=CC(C2=CC=C(CNC3CCCC3)S2)=CC=C1)NC(C1=C(C)C=CC(NC2CNC2)=C1)=O Chemical compound C[C@H](C1=CC(C2=CC=C(CNC3CCCC3)S2)=CC=C1)NC(C1=C(C)C=CC(NC2CNC2)=C1)=O PKMUHQIDVVOXHQ-HXUWFJFHSA-N 0.000 description 1
- 101710132601 Capsid protein Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- CCPHAMSKHBDMDS-UHFFFAOYSA-N Chetoseminudin B Natural products C=1NC2=CC=CC=C2C=1CC1(SC)NC(=O)C(CO)(SC)N(C)C1=O CCPHAMSKHBDMDS-UHFFFAOYSA-N 0.000 description 1
- 102000005598 Chondroitin Sulfate Proteoglycans Human genes 0.000 description 1
- 108010059480 Chondroitin Sulfate Proteoglycans Proteins 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 102000007665 Extracellular Signal-Regulated MAP Kinases Human genes 0.000 description 1
- 108010007457 Extracellular Signal-Regulated MAP Kinases Proteins 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- 208000031886 HIV Infections Diseases 0.000 description 1
- 208000037357 HIV infectious disease Diseases 0.000 description 1
- XLYOFNOQVPJJNP-ZSJDYOACSA-N Heavy water Chemical compound [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 1
- 102100022653 Histone H1.5 Human genes 0.000 description 1
- 101000899879 Homo sapiens Histone H1.5 Proteins 0.000 description 1
- 108010092694 L-Selectin Proteins 0.000 description 1
- 102100033467 L-selectin Human genes 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- OVRNDRQMDRJTHS-CBQIKETKSA-N N-Acetyl-D-Galactosamine Chemical compound CC(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-CBQIKETKSA-N 0.000 description 1
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 description 1
- 108010035766 P-Selectin Proteins 0.000 description 1
- 102100023472 P-selectin Human genes 0.000 description 1
- 208000037273 Pathologic Processes Diseases 0.000 description 1
- 102000003993 Phosphatidylinositol 3-kinases Human genes 0.000 description 1
- 108090000430 Phosphatidylinositol 3-kinases Proteins 0.000 description 1
- 102000001938 Plasminogen Activators Human genes 0.000 description 1
- 108010001014 Plasminogen Activators Proteins 0.000 description 1
- 102000002727 Protein Tyrosine Phosphatase Human genes 0.000 description 1
- 102000016611 Proteoglycans Human genes 0.000 description 1
- 108010067787 Proteoglycans Proteins 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical class [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- IOSLINNLJFQMFF-XMMPIXPASA-N [(2R)-1-[[4-[[3-[(4-fluorophenyl)methylsulfanyl]phenoxy]methyl]phenyl]methyl]pyrrolidin-2-yl]methanol Chemical compound FC1=CC=C(CSC=2C=C(OCC3=CC=C(CN4[C@H](CCC4)CO)C=C3)C=CC=2)C=C1 IOSLINNLJFQMFF-XMMPIXPASA-N 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000002491 angiogenic effect Effects 0.000 description 1
- 230000002424 anti-apoptotic effect Effects 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 210000001188 articular cartilage Anatomy 0.000 description 1
- 210000003050 axon Anatomy 0.000 description 1
- IVRMZWNICZWHMI-UHFFFAOYSA-N azide group Chemical group [N-]=[N+]=[N-] IVRMZWNICZWHMI-UHFFFAOYSA-N 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 239000003637 basic solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- SAGINAGERRNGGV-UHFFFAOYSA-N benzyl n-(2-hydroxyethyl)carbamate Chemical compound OCCNC(=O)OCC1=CC=CC=C1 SAGINAGERRNGGV-UHFFFAOYSA-N 0.000 description 1
- 239000012148 binding buffer Substances 0.000 description 1
- 230000033558 biomineral tissue development Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 208000029028 brain injury Diseases 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 230000032341 cell morphogenesis Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- DZRJLJPPUJADOO-UHFFFAOYSA-N chaetomin Natural products CN1C(=O)C2(Cc3cn(C)c4ccccc34)SSC1(CO)C(=O)N2C56CC78SSC(CO)(N(C)C7=O)C(=O)N8C5Nc9ccccc69 DZRJLJPPUJADOO-UHFFFAOYSA-N 0.000 description 1
- 239000007806 chemical reaction intermediate Substances 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 229940126545 compound 53 Drugs 0.000 description 1
- 229940125900 compound 59 Drugs 0.000 description 1
- 229940126179 compound 72 Drugs 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- JZCCFEFSEZPSOG-UHFFFAOYSA-L copper(II) sulfate pentahydrate Chemical compound O.O.O.O.O.[Cu+2].[O-]S([O-])(=O)=O JZCCFEFSEZPSOG-UHFFFAOYSA-L 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 230000002222 downregulating effect Effects 0.000 description 1
- 230000007783 downstream signaling Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 210000003989 endothelium vascular Anatomy 0.000 description 1
- 125000004494 ethyl ester group Chemical group 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 150000008275 galactosamines Chemical class 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 238000004896 high resolution mass spectrometry Methods 0.000 description 1
- 210000003630 histaminocyte Anatomy 0.000 description 1
- 239000012456 homogeneous solution Substances 0.000 description 1
- 208000033519 human immunodeficiency virus infectious disease Diseases 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 125000005524 levulinyl group Chemical group 0.000 description 1
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 210000000350 mc(t) Anatomy 0.000 description 1
- GDOPTJXRTPNYNR-UHFFFAOYSA-N methyl-cyclopentane Natural products CC1CCCC1 GDOPTJXRTPNYNR-UHFFFAOYSA-N 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 239000003226 mitogen Substances 0.000 description 1
- YCJZWBZJSYLMPB-UHFFFAOYSA-N n-(2-chloropyrimidin-4-yl)-2,5-dimethyl-1-phenylimidazole-4-carboxamide Chemical compound CC=1N(C=2C=CC=CC=2)C(C)=NC=1C(=O)NC1=CC=NC(Cl)=N1 YCJZWBZJSYLMPB-UHFFFAOYSA-N 0.000 description 1
- 210000000944 nerve tissue Anatomy 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 210000002241 neurite Anatomy 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 230000023362 neuron cell-cell adhesion Effects 0.000 description 1
- 230000007996 neuronal plasticity Effects 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000011164 ossification Effects 0.000 description 1
- 201000008968 osteosarcoma Diseases 0.000 description 1
- 239000005022 packaging material Substances 0.000 description 1
- 230000009054 pathological process Effects 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 229940127126 plasminogen activator Drugs 0.000 description 1
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 1
- 229920000447 polyanionic polymer Polymers 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108020000494 protein-tyrosine phosphatase Proteins 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 239000012146 running buffer Substances 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 230000037390 scarring Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 150000003384 small molecules Chemical group 0.000 description 1
- 239000007974 sodium acetate buffer Substances 0.000 description 1
- BEOOHQFXGBMRKU-UHFFFAOYSA-N sodium cyanoborohydride Chemical compound [Na+].[B-]C#N BEOOHQFXGBMRKU-UHFFFAOYSA-N 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 208000020431 spinal cord injury Diseases 0.000 description 1
- 125000000542 sulfonic acid group Chemical group 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- JRMUNVKIHCOMHV-UHFFFAOYSA-M tetrabutylammonium bromide Chemical compound [Br-].CCCC[N+](CCCC)(CCCC)CCCC JRMUNVKIHCOMHV-UHFFFAOYSA-M 0.000 description 1
- CZDYPVPMEAXLPK-UHFFFAOYSA-N tetramethylsilane Chemical compound C[Si](C)(C)C CZDYPVPMEAXLPK-UHFFFAOYSA-N 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- ZBZJXHCVGLJWFG-UHFFFAOYSA-N trichloromethyl(.) Chemical compound Cl[C](Cl)Cl ZBZJXHCVGLJWFG-UHFFFAOYSA-N 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
- A61P31/18—Antivirals for RNA viruses for HIV
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
- C07H15/20—Carbocyclic rings
- C07H15/203—Monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems
-
- 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
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
-
- 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
- C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
- C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
- C08B37/0069—Chondroitin-4-sulfate, i.e. chondroitin sulfate A; Dermatan sulfate, i.e. chondroitin sulfate B or beta-heparin; Chondroitin-6-sulfate, i.e. chondroitin sulfate C; Derivatives thereof
-
- 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)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Veterinary Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Polymers & Plastics (AREA)
- Materials Engineering (AREA)
- Neurosurgery (AREA)
- Neurology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Hospice & Palliative Care (AREA)
- Crystallography & Structural Chemistry (AREA)
- AIDS & HIV (AREA)
- Tropical Medicine & Parasitology (AREA)
- Psychiatry (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Dermatology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一系列如式所示的硫酸软骨素寡糖，其制备方法和生物活性，各中间体及合成方法。本发明的关键点在于：合成路线新颖，运用了糖苷化速率和综合产率都明显优于传统的“先糖苷化‑后氧化”策略，并应用硫代乙酸和1,3‑丙二硫醇解决了寡糖上叠氮还原的问题，收率和反应速率都在可接受的范围内，为今后的衍生多样化提供了实验依据，而且相关探索和发现有助于加深对CS‑E以及寡糖合成的了解，有助于未来CS‑E的构效关系和药理机制的研究，还有可能发现具有更优良药理活性的候选物。
Description
技术领域
本发明属于医药技术领域。本发明涉及一系列硫酸软骨素寡糖以及他们的合成方法、相关中间体合成方法，和此类化合物的生物活性。
背景技术
硫酸软骨素属于糖胺聚糖，是一种结构复杂，多阴离子的线性多糖，可通过共价键与蛋白质形成蛋白聚糖，以硫酸软骨素蛋白的形式广泛的存在于各种动物组织的细胞表面和细胞外基质，在骨、肌腱、血管、神经组织和软骨中特别丰富，尤其在人类结肠癌细胞和老年人的关节软骨中有很高的含量。硫酸软骨素蛋白参与很多生理活动，比如细胞分裂，形态构建和神经元可塑性；同时还参与了多种病理过程，包括骨骼疾病，大脑损伤后的神经元胶质瘢痕形成，细菌病毒感染等。
很多实验表明，硫酸软骨素蛋白的大部分功能依赖于糖链来执行，核心蛋白的部分只是作为骨架起支撑作用。而硫酸软骨素的结构是由葡萄糖醛酸和N-乙酰半乳糖胺以β-1,4，β-1,3糖苷键交替连接形成，根据O-硫酸化的位置和数目不同可分为多个亚型，常见的天然亚型有CS-A，CS-C，CS-D，CS-E，CS-K，CS-L和CS-M，尽管硫酸软骨素亚型的结构非常相似，只是磺酸类型不同，但是在活性上却存在较大差异，已有研究表明，硫酸软骨素特异性的序列可以编码功能信息，其重要的生理活性可能糖链上的磺酸基序列有关，这种序列可能根据组织和年龄不同而被严格控制，被称为磺酸密码。磺酸密码与硫酸软骨素的生物活性紧密相关，至今还未完全解译。
其中4,6-位二硫酸化的硫酸软骨素E与其他亚型相比具有很多特有的活性，例如参与骨髓分化成肥大细胞和粘膜肥大细胞的过程；与肝素结合因子相互作用，包括肝素结合细胞因子、L-选择素、P-选择素、CD44和趋化因子；抑制人类C1q因子；神经突的延长；抑制神经元细胞的粘着；修复脊髓损伤；骨的形成和生物矿，以及在很低的浓度时阻止HIV入侵细胞，还有报道利用其治疗人类关节炎。
但是硫酸软骨素的天然资源并不丰富，很难从自然界中获得结构确定的硫酸软骨素寡糖。为了解决硫酸软骨素的来源，通过化学合成的方法得到确定长度的硫酸软骨素寡糖是一种比较好的手段，而且得到结构确定的硫酸软骨素寡糖和衍生物，是对其准确的药理研究和构效关系研究不可或缺的基础，因此硫酸软骨素寡糖的化学合成收到了越来越多的重视。
肝素结合细胞因子(Midkine)选自一类新的结合肝素生长因子，是一种小分子的蛋白质，其作用的受体为硫酸软骨素蛋白多糖和蛋白质酪氨酸磷酸酶，下游的信号系统为细胞外信号调节激酶和PI3激酶，具有多种生物学功能：MK促进各种细胞的生长、存活和迁移，并可以促进神经轴突的生长和各种胚胎神经元的存活，因此与很多神经病变性疾病密切相关；经MK预处理的细胞可抑制HIV的黏附，预防HIV感染，且呈剂量依赖性；MK为一种有丝分裂原，与肿瘤的发生密切相关，研究发现MK在多种肿瘤组织和肿瘤患者的血清中的水平都明显升高，并可作为骨肉瘤的一种预后标志，因此为肿瘤标记和治疗的目标，成为潜在的抗肿瘤药物作用的新靶点；MK还有较强的促血管生成特性，已证实，其可促进内皮细胞增殖并提高血管的密度；此外，MK可通过上调纤维蛋白溶酶原激活剂的表达和下调该激活剂抑制因子的表达，增强血管内皮中纤溶酶的水平，增强纤溶作用；MK还可作为一种抗细胞凋亡的因子，通过抑制凋亡而保护细胞。因此Midkine蛋白与HIV、神经病变性疾病和肿瘤等多种疾病的发生和治疗密切相关。20来年，国外几个实验室在硫酸软骨素寡糖的全合成和衍生化方面做了很多研究工作，详细报道的硫酸软骨素寡糖的全合成路线有四条。一般以葡萄糖和半乳糖胺两个单糖或者葡萄糖-半乳糖胺二糖作为起始原料，合成的序列为GalN-GlcA和GlcA-GalN两种，通过一系列的选择性保护、糖基化、脱保护和官能团的转化，最终合成出不同长度的硫酸软骨素寡糖，目前报道的最长的是硫酸软骨素八糖。
从单糖开始合成GlcA-GalN序列的硫酸软骨素寡糖
2004年和2006年，Hsieh-Wilson实验室从单糖开始全合成了GlcA-GalN序列的二糖和四糖。他们首先以葡萄糖和半乳糖胺两个单糖为起始原料，分别合成了供体和受体，经过糖基化反应选择性的得到了GlcA-GalN序列的β连接的关键二糖前体1(74％)。二糖前体的非还原端和还原端保护基都可以选择性的进行脱除分别得到二糖的受体2和供体3，经过糖基化反应得到了β连接的四糖前体4(31％)，他们的合成策略为先氧化后糖苷化，糖苷化反应收率不高，而且糖链延长后收率明显降低。最后经过一系列的去保护、硫酸化和皂化反应得到了目标二糖5(51％)和四糖6(17％)。
2014年，Pedro M.Nieto课题组用N-TCP和N-TFA两种不同的半乳糖胺来探索合成CS-E寡糖。首先他们尝试了N-TCP保护的半乳糖胺，他们从单糖原料开始，合成了GlcA-GalN序列的二糖7(54％)，然而当他们想将二糖偶联得到四糖时却失败了，尝试反序连接依然没有成功，他们推测可能是由于TCP基团的体积过大影响了糖苷化反应。
接下来他们尝试了N-TFA保护的半乳糖胺，同样从单糖原料开始得到了GlcA-GalN序列的二糖8(91％)，这一次他们顺利的得到了GlcA-GalN序列的四糖9(70％)。而且顺利的在碱性条件下把TFA脱除，并且把氨基乙酰化，最终得到了过硫酸化的硫酸软骨素衍生物10。
2015年，Yasuo Suda课题组报道合成了一系列不同硫酸化的硫酸软骨素二糖和四糖，包括CS-E。他们也是采用了先氧化后糖苷化的合成策略，首先从单糖开始得到了2-位分别为三氯乙氧基甲酰基Troc保护的受体和2-位苯甲酰基Bz保护的供体，单糖经过偶联反应得到GlcA-GalN序列的二糖前体11。接下来他们为了后续的生物活性测定将二糖的端基转化为全苄基保护的葡萄糖，得到了三糖并转化为关键的三糖中间体12。经过一系列的去保护和硫酸化得到了硫酸软骨素E二糖和15个硫酸软骨素E类似物。
同年他们又在上面工作的基础上合成了硫酸软骨素E四糖和四个衍生物。他们对之前的二糖进行了结构改造，把半乳糖胺单元4,6-位的苄叉脱除，转化成四位Bn保护、六位Lev保护和双Lev保护的供体，然后对三糖也进行了类似的改造得到了三个受体。经过糖基化反应得到了四个五糖中间体13-16，最后经过去保护和硫酸化得到了硫酸软骨素E和四个四糖衍生物。
从单糖开始合成GalN-GlcA序列的硫酸软骨素寡糖
Jun-ichi-Tamura课题组从1998年开始对硫酸软骨素寡糖的合成开展了一系列研究，并且报道了GalN-GluA序列硫酸软骨素寡糖的第一条全合成路线。在这条路线中，半乳糖胺供体的2-位氨基采用了非参与性的叠氮结构，在三氟化硼乙醚/甲苯的条件下两个单糖进行偶联，可以选择性的生成β构型的二糖17。他们在二糖水平对6-位进行脱保护、氧化和甲酯化，把葡萄糖基转化为葡萄糖醛酸，得到了硫酸软骨素E二糖重复单元18，并延长了糖链。然而在六糖前体用Lindlar试剂将叠氮还原乙酰化时，收率不到10％，推测可能是由于分子中的叠氮数目过多导致收率过低。因此他们调整了合成策略，在二糖时先将叠氮还原乙酰化得到新的二糖中间体19，然后再延长糖链。得到了不同长度的硫酸软骨素寡糖前体，经过转化可得到相应的硫酸软骨素E寡糖。目前，作者使用该策略合成了硫酸软骨素E八糖。他们在合成四糖的时候发现，受体邻位为乙酰氨基时与化学惰性的叠氮不同，乙酰氨基比较活泼，在糖苷化的过程中乙酰氨基的羰基先亲核进攻供体快速生成一个动力学稳定的反应中间体，然后经过较长的反应时间后，经过SN1反应缓慢转化为热力学稳定的四糖。这导致糖苷化反应速度明显变慢，而且现象复杂。
2016年，Pedro M.Nieto课题组报道了第四条硫酸软骨素四糖的全合成路线，序列为GalN-GluA。与之前作者进行的相反序列的硫酸软骨素前体合成的研究的保护策略基本一致，较多的采用富电子集团保护，富电子集团的引入虽然可以促进糖的反应活性，但是却导致合成路线较长而且收率较低。采用该路线最终作者合成了硫酸软骨素E四糖20。
以天然硫酸软骨素二糖骨架为起始原料合成硫酸软骨素寡糖
2009年，Jacquinet课题组报道了一种新颖的硫酸软骨素寡糖的全合成思路。他们用酸降解牛源的硫酸软骨素聚糖，得到了单一的硫酸软骨素二糖。随后酯化得到二糖21，该二糖可以通过重结晶提纯。据报道，通过该制备方法，可轻易制备50-100g二糖原料。首先，他们应用了一系列的保护基策略，得将二糖21转化为二糖单元25。然后，他们利用该二糖单元延长得到了硫酸软骨素四糖、六糖前体26，糖苷化收率分别为71％和68％。并进一步将得到的硫酸软骨素前体转化为相应的硫酸软骨素寡糖，总收率分别为77％(四糖)和69％(六糖)。
综上所述，已报到的硫酸软骨素寡糖的合成路线中，二糖中间体都是氧化态的糖醛酸，即羧酸在单糖或二糖上引入，没有人尝试过在寡糖上引入羧基。本课题组的杨爽等人在前期的实验中发现，糖苷化反应中非氧化态的葡萄糖供体在反应收率和立体选择性方面都比糖醛酸供体有明显的优势，如果这一规律可以在硫酸软骨素的寡糖合成中应用，将大大提高糖苷化反应的效率。因此我们另辟新径，探索“先糖苷化-后氧化”策略在寡糖上的应用：以非氧化态的二糖单元为基础，合成葡萄糖基6位用对甲氧基苄醚保护的寡糖前体，然后脱除伯醇的保护，再将其氧化，最终转化为羧酸甲酯。最终构建出一条高效率的硫酸软骨素寡糖全合成路线，并得到了一系列硫酸软骨素寡糖以及中间体。
本专利申请保护的内容分为两个部分，第一部分是：全保护硫酸软骨素寡糖全合成路线。其优势为合成效率高，以葡萄糖和氨基半乳糖为原料，设计并合成二糖buildingblock，然后延长该二糖building block，得到系列硫酸软骨素E寡糖前体。在寡糖上构建糖醛酸单元，在糖苷化速率和综合产率方面都优于传统的“先氧化-后糖苷化”策略。应用含硫试剂，在寡糖上选择性还原叠氮，收率和反应速率都在可接受的范围内，解决了叠氮在寡糖上还原的问题，并为未来氨基衍生化奠定了基础。第二部分是：全保护硫酸软骨素寡糖转化为相应的亚型。将第一部分得到的部分寡糖前体，经过保护基的转换，得到硫酸软骨素O、A、C、E的寡糖。
发明内容
本发明解决的技术问题是提供一系列硫酸软骨素寡糖及其药学上可接受的盐，并提供寡糖及其相关中间体的合成方法，以及其应用。
为解决本发明的技术问题，本发明提供如下技术方案：
本发明技术方案的第一方面是提供一系列的全保护硫酸软骨素寡糖，具有以下的结构式：
其中，n＝1,2,3,4。R选自对甲基苯硫基(STol)或对甲氧基苯酚基(OMP)。
以及一系列不同亚型的硫酸软骨素寡糖，具有以下的结构式：
其中，n＝1,2,3，R1选自羟基(OH)或，R2选自羟基(OH)或硫酸钠基(OSO3Na)，R3选自对甲氧基苯酚基(OMP)P或苄氧羰基氨基乙基(OCH2CH2NHCbz)。
第一部分具体化合物列出如下：
化合物27
化合物28
化合物29
化合物30
化合物31
化合物32
化合物33
第二部分具体化合物列出如下：化合物34
化合物35
化合物36
化合物37
化合物38
化合物39
化合物40
本发明技术方案的第二方面是提供第一方面所述的硫酸软骨素寡糖的制备方法。其中起始原料为葡萄糖和氨基半乳糖，采用文献报道或化学上可行的反应方法，得到适当的中间体作为发明的原料，其结构式如下：
其中，Ph为苯基，Lev为乙酰丙酰基，N3为叠氮，Bz为苯甲酰基，STol为对甲基苯硫基，PMB为对甲氧基苄基。
本发明中原料的合成方法的反应式如下：
反应试剂和条件：a.乙酸酐，醋酸钠，90℃，2.5小时，94％；b.对甲基苯硫酚，三氟化硼乙醚络合物，二氯甲烷，40℃，8小时，71％；c.甲醇钠，甲醇，室温，1小时；d.对甲氧基苯甲醛二甲缩醛，对甲苯磺酸一水合物，N,N-二甲基甲酰胺，40℃，减压，6小时，89％；e.苯甲酰氯，三乙胺，二氯甲烷，室温过夜，70％；f.氰基硼氢化钠，三氟乙酸，分子筛，N,N-二甲基甲酰胺，室温，2天，93％；
反应试剂和条件：a..1)叠氮化钠，三氟甲磺酸酐，二氯甲烷，水，0℃，3小时；2)五水合硫酸铜，碳酸钾，甲醇/二氯甲烷/水，室温，24小时；3)甘氨酸,24小时4)乙酸酐，吡啶，4-二甲氨基吡啶，室温过夜，四步收率81％；b.对甲苯硫酚，三氟化硼乙醚络合物，二氯甲烷，40℃，过夜，75％；c.甲醇钠，甲醇，室温，30分钟；d.苯甲醛二甲缩醛对甲苯磺酸一水合物，N,N-二甲基甲酰胺，40℃，减压，2小时；e.乙酰丙酸，1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐，4-二甲氨基吡啶，二氯甲烷，室温，2小时，三步收率87％；f.碘代丁二酰亚胺，三氟乙酸，二氯甲烷/水,0℃,3小时,31％；g.三氯乙腈，1,8-二氮杂二环十一碳-7-烯，0℃，4小时，86％。
反应试剂和条件：a.三氟化硼乙醚，甲苯，-60℃，30分钟，96％；
本发明化合物合成方法包括将葡萄糖一水合物和氨基半乳糖盐酸盐的不同位点进行不同的保护，在对端基或4位进行选择性脱除后，进行糖苷化反应，得到适当的中间体作为发明的原料。
本发明技术方案的第三方面是提供了第一方面所述硫酸软骨素寡糖及其药学上可接受的盐在制备预防或治疗与肝素结合细胞因子(Midkine)相关疾病药物中的应用。所述的与肝素结合细胞因子(Midkine)相关疾病包括艾滋病、神经病变性疾病、肿瘤。
从发明具体实施例以及实验例的测定的结果中可以看出化合物34～36,40与肝素结合细胞因子(Midkine)相互作用具有一定的亲和力。因此第一方面所述的化合物可能会对肝素结合细胞因子(Midkine)生物学功能的发挥有一定的影响，从而发挥药理作用，经进一步研究可能会成为相应疾病例如HIV、神经病变性疾病和肿瘤等新的治疗药。
有益技术效果
本发明中的合成路线高效新颖，运用了糖苷化速率和综合产率都明显优于传统的“先糖苷化-后氧化”策略，并应用硫代乙酸和1,3-丙二硫醇解决了寡糖上叠氮还原的问题，收率和反应速率都在可接受的范围内，与以往的路线相同，随着糖链的延长，糖苷化收率逐渐降低，但合成至八糖收率依然高于70％；与糖链延长相关的反应，也都快速、高收率的进行，相应收率为81％～87％；综合来看我们构建的二糖单元具备高转化和延长效率，二糖、四糖和六糖合成相关步骤的综合收率为68％、77％和38％，而之前文献中报道的综合收率分别为12％、61％和23％。这都得益于“先糖苷化后氧化”策略的优势。端基衍生化的产品，为今后的衍生多样化提供了实验依据，而且相关探索和发现有助于加深对CS-E以及寡糖合成的了解，有助于未来CS-E的构效关系和药理机制的研究，还有可能发现具有更优良药理活性的候选物。
从发明具体实施例以及实验例的测定的结果中可以看出化合物34～36,40与肝素结合细胞因子(Midkine)相互作用具有一定的亲和力。
具体实施方式
通过下面的实施例可以对本发明进行进一步的描述，然而，本发明的范围并不限于下述实施例。本领域的专业人员能够理解，在不背离本发明的精神和范围的前提下，可以对本发明进行各种变化和修饰。本发明对试验中所使用到的材料以及试验方法进行一般性和/或具体的描述。虽然为实现本发明目的所使用的许多材料和操作方法是本领域公知的，但是本发明仍然在此作尽可能详细描述。
对于以下全部实施例，可以使用本领域技术人员已知的标准操作和纯化方法。除非另有说明，所有温度以℃(摄氏度)表示。化合物的结构是通过核磁共振谱(NMR)和/或质谱(MS)来确定的。
制备例部分
化合物的结构是通过核磁共振氢谱(1H NMR)、核磁共振碳谱(13C NMR)及质谱(MS)来确定的。核磁共振氢谱及碳谱位移(δ)以百万分之一(ppm)的单位给出。核磁共振氢谱用Mercury-300、Mercury-400、Mercury-500或Mercury-600型核磁共振仪测定，氘代氯仿(CDCl3)或重水(D2O)或氘代二甲基亚砜(DMSO-d6)作溶剂，四甲基硅烷(TMS)为内标。
高分辨质谱采用Agilent 1100series LC/MSD trap mass spectrometer液质联用仪或Q-Trap LC/MS/MS system(Applied Biosystems/MDS Sciex,USA)液质联用仪测定。
柱层析一般使用160～200目硅胶为载体。
无水溶剂均通过标准方法处理。其它试剂均为市售分析纯。
其中，
制备例1
本发明中原料47的合成方法的反应式如下：
将葡萄糖的一水合物(50g,253mmol)和乙酸钠(40g,410mmol)溶于360mL乙酸酐中，将反应液加热到90℃，搅拌2.5小时，TLC监测反应完全。将反应液倒入2.7L冰水中，搅拌3小时，白色固体析出。过滤，烘干，得到白色结晶性固体化合物42(92.57g，收率94％)。
1H NMR(CDCl3,400MHz):δ5.65(d,J＝8.4Hz,1H,H-1),5.19(t,J＝9.6Hz,1H,H-4),5.09-5.04(m,2H,H-3,H-2),4.23(dd,J＝12.4,4.4Hz,1H,H-6a),4.04(dd,J＝12.4,1.6Hz,1H,H6b),3.80-3.77(m,1H,H-5),2.05(s,3H,CH3-OAc),2.02(s,3H,CH3-OAc),1.97(s,6H,CH3-OAc),1.95(s,3H,CH3-OAc)；13C NMR(CDCl3,100MHz):δ170.80(CO-OAc),170.29(CO-OAc),169.58(CO-OAc),169.44(CO-OAc),169.15(CO-OAc),91.90(C-1),72.99,72.92,70.43,67.95,61.65,21.02(CH3-OAc),20.91(CH3-OAc),20.77(CH3-OAc).HRMS(ESI)m/z:413.1039[M+Na]+.Calcd.for C16H22NaO11 413.1060.
化合物42(86.14g,221mmol)溶解于345mL无水二氯甲烷中，冰水浴泠却至0℃，缓慢滴加BF3.Et2O(56.1mL,442mmol)，反应升至室温并加热回流，8小时后，反应用饱和碳酸氢钠水溶液淬灭，二氯甲烷萃取，有机相用饱和食盐水洗涤、干燥并旋干，得到黄色粗产品。用乙醇重结晶，得到白色固体(64.74g，收率是47％)。滤液浓缩后柱层析(乙酸乙酯：石油醚＝1：5)，得到化合物43(33.93g，收率24％)。
1H NMR(CDCl3,400MHz):δ7.36(d,J＝8.0Hz,2H,ArH-SPhMe),7.10(d,J＝8.0Hz,2H,ArH-SPhMe),5.18(t,J＝9.4Hz,1H,H-4),4.99(t,J＝9.8Hz,1H,H-3),4.90(t,J＝9.6Hz,1H,H-2),4.61(d,J＝10.0Hz,1H,H-1),4.21-4.13(m,2H,H-6a,b),3.69-3.67(m,1H,H-5),2.32(s,3H,CH3-SPhMe),2.06(s,3H,CH3-OAc),2.05(s,3H,CH3-OAc),1.99(s,3H,CH3-OAc),1.96(s,3H,CH3-OAc)；13C NMR(CDCl3,100MHz):δ170.73(CO-OAc),170.34(CO-OAc),169.56(CO-OAc),169.41(CO-OAc),138.97(ArC-SPhMe),134.03(2C,ArC-SPhCH3),129.87(2C,ArC-SPhMe),127.74(ArC-SPhMe),85.98(C-1),75.94,74.21,70.11,68.39,62.32,21.39(CH3-SPhMe),20.97(CH3-OAc),20.93(CH3-OAc),20.79(CH3-OAc),20.78(CH3-OAc).ESI-MS m/z:476.88[M+Na]+.Calcd.for C21H26O9SNa 477.12.
化合物43(86.14g,221mmol)溶于149mL二氯甲烷和324mL甲醇中，加入甲醇钠(4.09g,76mmol)。室温搅拌1小时后，TLC监测反应完全，加入酸性树脂DOWEX 50X8-200，搅拌1小时。待PH为中性时，过滤除去树脂，浓缩滤液得到黄色固体化合物44(37.78g，收率93％)。该化合物直接用于下一步反应。
1H NMR(CD3OD,400MHz):δ7.45(d,J＝7.2Hz,2H,ArH-SPhMe),7.10(d,J＝7.2Hz,2H,ArH-SPhMe),4.53(d,J＝10.0Hz,1H,H-1),3.89(d,J＝12.0Hz,1H,H-6a),3.67(d,J＝12.0Hz,1H,H-6b),3.41-3.30(m,H-3,H-4,overlapped with solvent signal),3.21(t,J＝9.0Hz,1H,H-2),2.28(s,3H,CH3-SPhMe)；13C NMR(CD3OD,100MHz):δ137.60(ArC-SPhMe),132.34(2C,ArC-SPhMe),129.95(ArC-SPhMe),129.44(2C,ArC-SPhMe),88.42(C-1),80.73,78.41,72.48,70.14,61.72,20.09(CH3-SPhMe).
化合物44(5g,17.5mmol)溶于17mL无水DMF中，加入对甲氧基苯甲缩醛(133mg,0.7mmol)和对甲苯磺酸的一水合物(133mg,0.7mmol)，40℃减压反应6小时，TLC监测反应完全。反应液用三乙胺淬灭，乙酸乙酯稀释，有机相用饱和食盐水洗涤。干燥旋干有机相，柱层析(乙酸乙酯：石油醚＝1：5)，得到白色固体化合物45(5.83g，收率89％)。
1H NMR(CDCl3,500MHz):δ7.42(d,J＝8.0Hz,2H,ArH-SPhMe),7.39(d,J＝8.5Hz,2H,ArH-PMB),7.14(d,J＝8.0Hz,2H,ArH-SPhMe),6.87(d,J＝8.5Hz,2H,ArH-PMB),5.45(s,1H,CH-PMB),4.53(d,J＝9.5Hz,1H,H-1),4.33(dd,J＝10.5,2.5Hz,1H,H-6a),3.78-3.71(m,5H,H-6b,H-4,OCH3-PMB),3.66-3.61(m,3H,H-2,H-3,H-5),3.13(brs,1H,OH),2.91(brs,1H,OH),2.35(s,3H,CH3-SPhMe)；13C NMR(CDCl3,125MHz):δ160.46(ArC-PMB),139.02(ArC-SPhMe),133.84(2C,ArC-SPhMe),130.10(2C),129.55,127.85(2C),127.51,113.92(2C,ArC-PMB),102.03(CH-PMB),88.84(C-1),80.35,74.69,72.64,70.68,68.74,55.52(OCH3-PMB),21.41(CH3-SPhMe).ESI-MS m/z:404.71(M+H)+.Calcd.for C21H25O6S 404.13.
化合物45(49.28g,132mmol)溶于375mL吡啶中，0℃加入苯甲酰氯(37.9mL,329mmol)，自然升至室温并搅拌过夜。TLC监测反应完全，反应液倒入冰水中，白色固体析出，过滤得到粗产品，用二氯甲烷溶解，加硅胶拌样，柱层析(二氯甲烷：石油醚＝1：1)，得到白色固体化合物46(57.29g，收率70％)。
1H NMR(CDCl3,400MHz):δ7.98(d,J＝8.0Hz,2H,ArH-Bz),7.93(d,J＝8.4Hz,2H,ArH-Bz),7.53(t,J＝7.4Hz,1H,ArH-Bz),7.48(t,J＝7.4Hz,1H,ArH-Bz),7.42-7.32(m,8H,ArH-Bz,SPhMe and PMB),7.13(d,J＝8.0Hz,2H,ArH-SPhMe),6.83(d,J＝8.4Hz,2H,ArH-PMB),5.77(t,J＝9.6Hz,1H,H-3),5.50(s,1H,CH-PMB),5.45(t,J＝9.6Hz,1H,H-2),4.97(d,J＝10.0Hz,1H,H-1),4.44(dd,J＝10.4,4.8Hz,1H,H-6a),3.90-3.83(m,H-6b,2H,H-4),3.76-3.69(m,4H,OCH3-PMB,H-5),2.35(s,3H,CH3-SPhMe)；13C NMR(CDCl3,100MHz):δ165.81(CO-Bz),165.41(CO-Bz),160.30(ArC-PMB),138.97(ArC-SPhMe),133.95(2C,ArC-SPhMe),133.52(ArC-Bz),133.29(ArC-Bz),130.10(2C),130.02(2C,ArC-Bz),130.00(2C,ArC-Bz),129.63,129.49(ArC-Bz),129.46(ArC-Bz),128.62(2C),128.49(2C),128.17,127.68(2C),113.78(2C,ArC-PMB),101.68(CH-PMB),87.49(C-1),78.76,73.60,71.33,71.19,68.71,55.48(OCH3-PMB),21.44(CH3-SPhMe).ESI-MS m/z:612.89[M+H]+.Calcd.for C35H33O8SH613.19.
化合物46(10g,48.78mmol)溶于100mL无水DMF中，加入NaBH3CN(5.1g,243.88mmol)和分子筛(10g)，0℃滴加三氟乙酸(12.1mL,487.76mmol)，然后自然升至室温并搅拌三天。TLC监测反应完全，滤除分子筛，反应液用二氯甲烷稀释，倒入饱和碳酸氢钠水溶液中。水相用二氯甲烷萃取，有机相用饱和食盐水洗涤，干燥并旋干，柱层析(乙酸乙酯：石油醚＝1：10)，得到白色固体化合物47(11g，收率93％)。
1H NMR(CDCl3,400MHz):δ7.90-7.85(m,4H,ArH-Bz),7.45-7.18(m,10H,ArH-Bz,PMB and SphMe),6.98(d,J＝8.0Hz,2H,ArH-SphMe),6.81(d,J＝8.8Hz,2H,ArH-PMB),5.37(t,J＝9.2Hz,1H,H-3),5.30(t,J＝9.6Hz,1H,H-2),4.78(d,J＝9.6Hz,1H,H-1),4.46(s,2H,CH2-PMB),3.83(td,J＝3.2,9.6Hz,1H,H-4),3.77-3.76(m,2H,H-6),3.73(s,3H,OCH3-PMB),3.64-3.60(m,1H,H-5),3.17(d,J＝3.2Hz,1H,OH),2.24(s,3H,CH3-SphMe)；13C NMR(CDCl3,100MHz):δ167.64(CO-Bz),165.48(CO-Bz),159.63(ArC-PMB),138.71(ArC-SPhMe),133.76(CO-Bz),133.71(2C,ArC-SPhMe),133.52(CO-Bz),130.18(2C),130.04(2C),129.94(2C),129.84,129.77(2C),129.52,129.08,128.64(2C),128.62(2C),128.34,114.13(2C,ArC-PMB),86.52(C-1),78.75(C-5),77.95(CH2-PMB),73.71(C-3),71.11(C-2),70.33(C-4),69.89(C-6),55.51(OCH3-PMB),21.41(CH3-SPhMe).ESI-MS m/z:636.97[M+Na]+.Calcd.for C35H34O8SNa 637.19.
NaN3(59.52g,0.91mol)溶于192mL水和252mL二氯甲烷中，降温至0℃，剧烈搅拌，缓慢滴加三氟甲磺酸酐(31.2mL,0.18mol)，反应继续搅拌3小时，分离出有机相，水层用少量二氯甲烷萃取，有机相总体积约为340mL。有机相用饱和碳酸氢钠水溶液洗涤两次，得到叠氮试剂溶液。
半乳糖胺盐酸盐(20g,0.093mol)溶于150mL水中，加入碳酸钾(9.6g,69.46mmol)、CuSO4.5H2O(110mg,1.02mmol)、200mL甲醇和以上制备的TfN3溶液。然后再加入约280mL甲醇使得反应液成为均相。反应在室温搅拌24小时后，加入甘氨酸(35g,0.47mol)，继续搅拌24小时。后处理：过滤，滤液蒸干，所得粗产品用甲苯带水3次，得到黄色固体，溶于400mL吡啶中，加入乙酸酐(112mL,mmol)和催化量DMAP(42mg,mmol)，室温搅拌过夜。后处理：蒸除溶剂，用甲醇溶解、加入硅胶拌样，柱层析(乙酸乙酯：石油醚＝1：4)，得到无色浆状化合物49(28.16g，收率81％)。
1H NMR(CDCl3,400MHz)δ5.52(d,J＝8.4Hz,1H,H-1),5.33(d,J＝2.4Hz,1H,H-4),4.88(dd,J＝10.8,3.2Hz,1H,H-3),4.12-3.97(m,2H,H-6a,b,H-5),3.77(t,J＝8.8Hz,1H,H-2),2.14(s,3H,OCH3-OAc),2.11(s,3H,OCH3-OAc),2.01(s,3H,OCH3-OAc),1.98(s,3H,CH3-OAc)；13C NMR(CDCl3,100MHz)δ170.25(CO-OAc),169.89(CO-OAc),169.54(CO-OAc),168.51(CO-OAc),90.77(C-1),71.65,71.21,66.21,60.97,59.68(C-2),20.82(CH3-OAc),20.57(CH3-OAc),20.53(CH3-OAc),20.50(CH3-OAc).ESI-HRMS m/z:396.1002[M+Na]+,412.0728[M+K]+.Calcd.for C14H19N3NaO9396.1019,C14H19N3KO9412.0758.
将化合物49(49.6g,132.93mmol)溶于208ml无水二氯甲烷中，加入4-甲基苯硫酚(24.77g,199.4mmol)。在0℃搅拌下，缓慢滴加三氟化硼乙醚(67.53ml,531.76mmol)，继续搅拌30min，然后升温至40℃，搅拌过夜，后处理：饱和碳酸氢钠水溶液淬灭反应，并调PH为中性，二氯甲烷萃取，有机相干燥并旋干，柱层析(乙酸乙酯：石油醚＝1：6)，得到黄色浆状化合物50(45.19g，收率75.0％)。
50a:1H NMR(CDCl3,400MHz):δ7.41(d,J＝8.0Hz,2H),7.13(d,J＝9.2Hz,2H),5.61(d,J＝5.6Hz,1H,H-1),5.48(d,J＝2.4Hz,1H,H-4),5.18(dd,J＝11.2,2.8Hz,1H,H-3),4.77(t,J＝6.4Hz,1H,H-5),4.30(dd,J＝11.2,5.6Hz,1H,H-2),4.15(d,J＝6.8Hz,1H,H-6a),4.12(d,J＝6.8Hz,1H,H-6b),2.34(s,3H,CH3-STol),2.15(s,3H,CH3-OAc),2.06(s,3H,CH3-OAc),2.01(s,3H,CH3-OAc)；
50b:1H NMR(CDCl3,400MHz):δ7.51(d,J＝8.0Hz,2H),7.16(d,J＝8.8Hz,2H),5.34(d,J＝2.8Hz,1H,H-4),4.86(dd,J＝10.2,2.8Hz,1H,H-3),4.46(d,J＝10.0Hz,1H,H-1),4.09-4.07(d,2H,J＝6.4Hz,H-6a,b),3.87(t,J＝6.4Hz,1H,H-5),3.63(t,J＝10.0Hz,1H,H-2),2.36(s,3H,CH3-STol),2.09(s,3H,CH3-OAc),2.04(s,3H,CH3-OAc),2.03(s,3H,CH3-OAc)；
50ab(α/β)1:1.2:13C NMR(CDCl3,100MHz):δ170.47,170.44,170.07,170.02,169.81,169.67,139.01,138.50,134.08,133.23,130.04,129.85,128.65,127.31,87.40(α,C-1),86.78(β,C-1),74.41,73.09,70.21,67.61,67.51,66.62,61.84,61.65,59.40,58.24,21.31,21.25,20.78,20.74,20.72,20.65.HRMS m/z:460.1108[M+Na]+.Calcd.forC19H23N3NaO7S 460.1154.
将化合物50(45.19g,103.37mmol)溶于104mL二氯甲烷和234mL甲醇中，搅拌形成均一溶液，加入甲醇钠(2.96g,54.78mmol)，室温搅拌30分钟，加入酸性树脂，继续搅拌30分钟至PH为中性，过滤除去树脂，旋干滤液得到粗产品51.
将化合物51的粗产品溶于103mL无水DMF，加入苯甲缩醛(46.5mL,310.11mmol)，对甲苯磺酸一水合物(4.72g,24.81mmol)，40℃减压反应2小时，后处理：向反应液中加三乙胺至中性，加二氯甲烷稀释，有机相用饱和食盐水洗涤，干燥并旋干，得到黄色油状化合物52。
52a:1H NMR(CDCl3,400MHz):δ7.51-7.48(m,2H,ArH-STol),7.40-7.36(m,5H,ArH-benzylidene),7.11(d,J＝8.0Hz,2H,ArH-STol),5.67(d,J＝5.2Hz,1H,H-1),5.59(s,1H,CH-benzylidene),4.31(dd,J＝3.6,0.8Hz,1H,H-4),4.26-4.21(m,2H,H-2,H-5),4.17(dd,J＝12.4,5.2Hz,1H,H-6a),4.11(dd,J＝12.4,1.6Hz,1H,H-6b),4.00(td,J＝10.4,3.6Hz,1H,H-3),2.69(d,J＝10.0Hz,1H,OH),2.33(s,3H,CH3-STol)；13C NMR(CDCl3,100MHz)δ137.82,137.28,131.78,130.03,129.87,129.55,128.48,126.35,101.46(CH-benzylidene),87.78(C-1),75.25,69.65,69.30,63.67,61.54(C-2),21.21(CH3-STol).
52b:1H NMR(CDCl3,400MHz):δ7.62(d,J＝8.4Hz,2H,ArH-STol),7.43-7.36(m,5H,ArH-benzylidene),7.11(d,J＝8.0Hz,2H,ArH-STol),5.51(s,1H,CH-benzylidene),4.38(dd,J＝12.4,1.2Hz,1H,H-6a),4.36(d,J＝9.6Hz,1H,H-1),4.16(d,J＝3.6Hz,1H,H-4),4.02(dd,J＝12.4,1.6Hz,1H,H-6b),3.62(td,J＝9.6,3.6Hz,1H,H-3),3.49(d,J＝0.8Hz,1H,H-5),3.48(t,J＝9.6Hz,1H,H-2),2.51(d,1H,J＝9.6Hz,OH),2.36(s,3H,CH3-STol)；13CNMR(CDCl3,100MHz)δ138.86,137.51,134.92,129.90,129.60,128.37,126.70,126.43,101.53(CH-benzylidene),85.14(C-1),74.55,73.32,69.93,69.38,62.13(C-2),21.40(CH3-STol).HRMS m/z:400.1319[M+H]+,422.1138[M+Na]+.Calcd.for C20H22N3O4S400.1331,C20H21N3NaO4S 422.1150.
将化合物52的粗产品溶于752mL二氯甲烷，依次加入乙酰丙酸(30.0g,258.4mmol)，EDCI(49.5g,258.4mmol)，DMAP(3.79g,31.01mmol)，室温搅拌2小时。后处理：将反应液倒入水中，用二氯甲烷萃取，有机相用饱和食盐水洗涤，干燥并旋干，柱层析(乙酸乙酯：石油醚＝1：6至2:1)，得到黄色浆状化合物53(44.27g，三步收率86.8％)。
1H NMR(CDCl3,400MHz):δ7.61(d,J＝8.0Hz,2H,ArH-STol),7.43-7.36(m,5H,ArH-benzylidene),7.06(d,J＝8.4Hz,2H,ArH-STol),5.47(s,1H,CH-benzylidene),4.83(dd,J＝10.4,3.2Hz,1H,H-3),4.45(d,J＝9.6Hz,1H,H-1),4.34(dd,J＝12.8,1.6Hz,1H,H-6a),4.27(dd,J＝3.2Hz,1H,H-4),3.62(dd,J＝12.4,1.6Hz,1H,H-6b),3.79(t,J＝10.0Hz,1H,H-2),3.51(s,1H,H-5),2.72-2.69(m,2H,CH2-Lev),2.60-2.55(m,2H,CH2-Lev),2.33(s,3H,CH3-STol),2.06(s,3H,CH3-Lev)；13C NMR(CDCl3,100MHz)δ206.26(CO-Lev),172.03(CO-Lev),138.68,137.66,134.62,129.83,129.17,128.11,126.53,126.16,100.86(CH-benzylidene),85.23(C-1),73.89,72.61,69.54,69.14,58.32(C-2),37.80(CH2-Lev),29.63(CH3-Lev),28.05(CH2-Lev),21.26(CH3-STol).HRMS m/z:498.1684[M+H]+,520.1500[M+Na]+.Calcd.for C25H28N3O6S 498.1699,C25H27N3NaO6S 520.1518.
方法1：化合物53b(265.2mg,0.53mmol)溶于3mL无水二氯甲烷中，加入150μL水，NIS(144mg，0.64mmol)和三氟乙酸(20μL，0.26mmol)。室温剧烈搅拌5小时，TLC监测反应结束。后处理：反应液用饱和硫代硫酸钠水溶液淬灭，倒入水中，用二氯甲烷萃取，有机相干燥并旋干，柱层析(乙酸乙酯：石油醚＝2：5)，得到浅黄色浆状化合物54(108mg，收率51.7％)。
方法2：将化合物53a(48.27g,97.88mmol)溶于532.2mL二氯甲烷和26.5mL水中，降温至0℃，加入NIS(26.43g,117.46mmol)，三氟乙酸(3.65mL,48.94mmol)，0℃避光搅拌3小时。后处理：反应液用饱和硫代硫酸钠水溶液淬灭，倒入饱和碳酸氢钠溶液中，二氯甲烷萃取，有机相用饱和氯化钠溶液洗2次，干燥，柱层析(乙酸乙酯：石油醚＝1：5至1：2)，回收原料53a 21.07g，得到黄色浆状化合物54(11.92g，收率31.1％)。
54ab,(α/β)1:0.3:1H NMR(CDCl3,400MHz):δ7.52-7.50(m,2H),7.40-7.36(m,3H),5.54(s,1H,CH-benzylidene),5.47(d,J＝3.2Hz,Ha-1),5.38(dd,J＝10.8,3.2Hz,1H,Ha-3),4.77(dd,J＝10.8,3.2Hz,1H,Hb-3),4.63(d,J＝8.0Hz,1H,Hb-1),4.43(d,J＝3.2Hz,1H,Ha-4),4.32-4.28(m,2H,Hb-4,Hb-6a),4.22(dd,J＝12.4,1.2Hz,1H,Ha-6a),4.07-3.98(m,4H,Ha-2,Ha-5,Ha-6b,Hb-6b),3.87(dd,J＝11.8,8.0Hz,1H,Hb-2),3.45(s,1H,Hb-5),2.79-2.65(m,4H,CH2-Lev),2.12(s,3H,CH3-Lev)；13C NMR(CDCl3,100MHz)δ206.48(CO-Lev),172.30(CO-Lev),137.65,129.19,128.33,128.32,126.34,126.30,100.85(CH-benzylidene),96.53(Cb-1),92.88(Ca-1),73.58,72.73,72.44,69.73,69.08,66.69,62.62(C-2),62.05,58.05(C-2),38.01(CH2-Lev),29.82(CH3-Lev),28.28(CH2-Lev).HRMS m/z:414.1256[M+Na]+.Calcd.for C18H21N3NaO7 414.1277.
将化合物54(20.153g,51.53mmol)溶于504mL无水二氯甲烷，降温至0℃搅拌，依次加入CCl3CN(31mL,309.15mmol)，DBU(3.08mL,20.61mmol)，0℃搅拌4小时。后处理：加入三乙胺和硅胶拌样，柱层析(乙酸乙酯：石油醚＝1：15，5‰三乙胺)，得到白色固体化合物55(23.56g，收率85.6％)。
1H NMR(CDCl3,400MHz):δ8.77(s,1H,NH),7.53-7.51(m,2H,ArH-benzylidene),7.39-7.37(m,3H,ArH-benzylidene),5.58(d,J＝3.2Hz,1H,H-1),5.56(s,1H,CH-benzylidene),5.37(dd,J＝10.8,3.6Hz,1H,H-3),4.55(dd,J＝3.2Hz,1H,H-4),4.32-4.28(m,2H,H-6a,b),4.04(dd,J＝12.8,2.8Hz,1H,H-2),3.98(s,3H,H-5),2.80-2.66(m,4H,CH2-Lev),2.12(s,3H,CH3-Lev)；13C NMR(CDCl3,100MHz)δ206.35,172.18,160.68(C＝NH),137.39,129.29,128.33,126.28,100.87(CH-benzylidene),95.44(C-1),90.89(CCl3),72.91,69.93,68.80,64.91,56.95(C-2),37.88(CH2-Lev),29.77(CH3-Lev),28.18(CH2-Lev).
化合物55(978.8mg,1.83mmol)和化合物47(625.2mg,1.02mmol)用无水甲苯带水三次，然后将其溶于34mL无水甲苯中，加入2g新烘干的分子筛。氩气保护下室温搅拌30分钟，然后将反应瓶置于-60℃，搅拌15分钟后，加入BF3.Et2O(242μL，0.76M，0.183mmol)的二氯甲烷溶液，继续搅拌30分钟后，TLC监测反应结束。后处理：加入过量三乙胺淬灭反应，然后过滤分子筛并浓缩滤液，先后进行柱层析(乙酸乙酯：石油醚＝1：2)，重结晶(乙酸乙酯：石油醚＝1：5)，得到白色固体化合物27(963.1mg，收率96.3％)。
1H NMR(CDCl3,400MHz)δ7.95-7.93(m,2H,ArH-Bz),7.91-7.89(m,2H,ArH-Bz),7.50(t,J＝7.4Hz,1H,ArH-Bz),7.40-7.16(m,14H,ArH-Bz,PMB,SPhMe,benzylidene),7.06(d,J＝8.0Hz,1H,ArH-SPhMe),6.92(d,J＝8.6Hz,1H,ArH-PMB),5.69(t,J＝9.2Hz,1H,H1-3),5.35(t,J＝9.6Hz,1H,H1-2),5.21(s,1H,CH-benzylidene),4.85(d,J＝10.0Hz,1H,H1-1),4.63(d,J＝11.5Hz,1H,CH2-PMB),4.54(dd,J＝10.8,3.6Hz,1H,H2-3),4.51(d,J＝11.6Hz,1H,CH2-PMB),4.31(d,J＝8.0Hz,1H,H2-1),4.21(t,J＝9.6Hz,1H,H1-4),4.03(d,J＝3.4Hz,1H,H2-4),3.97(dd,J＝11.2,3.6Hz,1H,H1-6a),3.91(d,J＝10.4Hz,1H,H1-6b),3.82(s,3H,OCH3-PMB),3.76-3.71(m,2H,H2-2,H1-5),3.56(s,2H,H2-6a,b),2.90(s,1H,H2-5),2.71-2.68(m,2H,CH2-Lev),2.60-2.57(m,2H,CH2-Lev),2.32(s,3H,CH3-Lev),2.05(s,3H,CH3-SPhMe)；13C NMR(CDCl3,100MHz):δ206.18(CO-Lev),172.05(CO-Lev),165.71(CO-Bz),165.34(CO-Bz),159.44(ArC-PMB),138.46(ArC-SPhMe),137.66(ArC-benzylidene),133.72(2C,ArC-SphMe),133.23(ArC-Bz),132.76(ArC-Bz),130.36,129.97,129.94,129.78,129.75,129.67,129.56,128.96,128.43,128.40,128.13,127.99,126.62,113.94(2C,ArC-PMB),101.38(PhCH),100.78(C2-1),86.36(C1-1),79.25,75.42,74.64,73.24,72.72,72.29,70.98,68.10,67.96,66.19,60.63,55.42(OCH3-PMB),37.91(CH2-Lev),29.74(CH3-Lev),28.18(CH2-Lev),21.30(CH3-SPhMe).ESI-HRMS m/z:988.3295[M+H]+,1010.3027[M+Na]+.Calcd.for C53H54N3O14S 988.3326,C53H53N3NaO14S 1010.3146.
实施例1
化合物27(500mg,0.507mmol)溶于13mL二氯甲烷中，搅拌下加入水1.3mL，NIS(228mg,1.01mmol)，三氟乙酸(19μL，0.253mmol)。室温搅拌1小时，TLC监测反应完全。后处理：反应液用饱和硫代硫酸钠水溶液淬灭，倒入水中，二氯甲烷萃取，有机相干燥并旋干，得到白色泡沫状化合物56，直接用于下步反应。HRMS(ESI)m/z:904.2855[M+Na]+.Calcd.forC46H47N3NaO15 904.2905.
以上粗产品56溶于10mL无水二氯甲烷中，降温至0℃，搅拌下加入DBU(30μL，0.20mmol)和三氯乙腈(305μL，3.04mmol)，自然升至室温，30分钟后，TLC监测反应结束。后处理：加入少量三乙胺和硅胶拌样，柱层析(乙酸乙酯：石油醚＝1：5至1：2)，得到白色泡沫状化合物57(409.4mg，两步收率78.9％)。
1H NMR(CDCl3,500MHz)δ8.57(s,1H,NH),7.95(m,4H,ArH-Bz),7.47(t,J＝7.1Hz,1H,ArH-Bz),7.39–7.15(m,12H,ArH-Bz,PMB,benzylidene),6.92(d,J＝8.3Hz,2H,ArH-PMB),6.73(d,J＝3.0Hz,1H,H1-1),6.09(t,J＝9.8Hz,1H,H1-3),5.47(dd,J＝10.1,3.3Hz,1H,H1-2),5.23(s,1H,CH-benzylidene),4.66(d,J＝11.5Hz,1H,CH2-PMB),4.53(dd,J＝10.6,3.0Hz,1H,H2-3),4.46(d,J＝11.5Hz,1H,CH2-PMB),4.38(t,J＝9.5Hz,1H,H1-4),4.26(m,2H,H2-4,H1-6a),4.06(m,2H,H1-6b),3.82(s,5H,OCH3-PMB,H2-2,H1-5),3.58(s,2H,H2-6a,b),2.93(s,1H,H2-5),2.70(m,2H,CH2-Lev),2.60(m,2H,CH2-Lev),2.05(s,3H,CH3-Lev).13C NMR(125MHz,CDCl3)δ206.14(CO-Lev),172.00(CO-Lev),165.63(CO-Bz),165.59(CO-Bz),160.83(C＝NH),159.56(ArC-PMB),137.64(ArC-benzylidene),133.47(ArC-Bz),132.83(ArC-Bz),129.98,129.90,129.82,128.98,128.82,128.45,128.16,127.97,126.62,114.01(ArC-PMB),101.32(PhCH),100.81(C2-1),93.52(C1-1),90.92,74.87,73.25,72.99,72.90,72.22,70.98,70.40,68.08,67.20,66.21,60.43,55.39(OCH3-PMB),37.87(C-Lev),29.70(C-Lev),28.17(C-Lev).
化合物57(390mg,0.38mmol)和对甲氧基苯酚(56.7mg,0.46mmol)用无水甲苯带水三次，然后将其溶于5mL无水二氯甲烷中，加入800mg新烘干的分子筛。氩气保护下室温搅拌30分钟，然后将反应瓶置于-60℃，搅拌15分钟后，加入TMSOTf(103μL,0.37M,0.038mmol)的二氯甲烷溶液，继续搅拌30分钟后，TLC监测反应结束。后处理：加入过量三乙胺淬灭反应，然后过滤分子筛并浓缩滤液，柱层析(乙酸乙酯：石油醚＝1：2)，得到白色浆状化合物31(343.1mg，收率91.2％)。
1H NMR(CDCl3,500MHz)δ7.97-7.93(m,4H,ArH-Bz),7.45(t,J＝7.0Hz,1H,ArH-Bz),7.34–7.21(m,12H,ArH-Bz,PMB,benzylidene),6.94(d,J＝8.5Hz,2H,ArH-PMP),6.89(d,J＝8.0Hz,2H,ArH-PMB),6.74(d,J＝8.5Hz,2H,ArH-PMP),5.77(t,J＝9.0Hz,1H,H1-3),5.63(t,J＝9.0Hz,1H,H1-2),5.22(s,1H,CH-benzylidene),5.15(d,J＝8.0Hz,1H,H1-1),4.66(d,J＝11.5Hz,1H,CH2-PMB),4.57(dd,J＝10.5,2.5Hz,1H,H2-3),4.50(d,J＝11.5Hz,1H,CH2-PMB),4.34-4.30(m,2H,H2-1,H1-4),4.05(s,1H,H2-4),4.00(dd,J＝11.0,3.0Hz,1H,H2-6a),3.93(d,J＝11.0Hz,1H,H2-6b),3.88(d,J＝9.5Hz,1H,H1-5),3.78-3.74(m,4H,OCH3-PMB,H2-2),3.70(OCH3-PMP),3.57(brs,2H,H1-6a,b),2.91(s,1H,H2-5),2.68-2.66(m,2H,CH2-Lev),2.59-2.56(m,2H,CH2-Lev),2.03(s,3H,CH3-Lev)；13C NMR(125MHz,CDCl3)δ206.05(CO-Lev),171.87(COO-Lev),165.53(CO-Bz),165.24(CO-Bz),159.33(ArC-PMB),155.58(ArC-PMP),151.20(ArC-PMP),137.60(ArC-benzylidene),133.13(ArC-Bz),132.75(ArC-Bz),130.04,129.83,129.72,129.62,129.33,128.85,128.32,128.06,127.86(ArC-benzylidene),126.48(ArC-benzylidene),118.87(ArC-PMP),114.01(ArC-PMP),113.82(ArC-PMB),101.32(C2-1),100.81(C1-1),100.61(CH-benzylidene),75.50(C1-4),75.06(C1-5),73.27(C1-3),73.14(CH2-PMB),72.56(C2-3),72.16(C2-4),72.05(C1-2),67.95(C1-6),67.62(C2-6),66.09(C-5),60.57(C2-2),55.53(OCH3-PMP),55.23(OCH3-PMB),37.87(CH2-Lev),29.70(CH3-Lev),28.17(CH2-Lev).HRMS(ESI)m/z:988.3516[M+H]+,1010.3294[M+Na]+.Calcd.for C53H54N3O16 988.3504,C53H53N3NaO16 1010.3324.
化合物31(469mg,0.48mmol)溶于12mL二氯甲烷中，搅拌下加入水640μL，DDQ(130mg,0.57mmol)。室温剧烈搅拌3小时，TLC监测反应完全。后处理：反应液用二氯甲烷稀释，饱和碳酸氢钠水溶液洗涤两次，饱和食盐水洗涤一次，有机相干燥并旋干，柱层析(乙酸乙酯：石油醚＝1：1)得到白色泡沫状化合物58(357mg，收率87％)。
化合物58(600mg,0.69mmol)溶于4.5mL CH2Cl2/buffer(Na2CO3/NaHCO3,PH 9.5)(2/1)混合溶液中，室温剧烈搅拌，加入TEPMPO(0.1M,1.4mL,0.14mmol)的二氯甲烷溶液，碘苯二乙酸(557mg,1.73mmol)。室温搅拌4小时后，TLC监测反应结束。后处理：反应液用饱和硫代硫酸钠水溶液淬灭，然后倒入饱和食盐水中，乙酸乙酯萃取，有机相干燥和旋干，得到黄色浆状化合物59。
以上粗产品溶解于9mL DMF中，加入碘甲烷(345μL,5.54mmol)和碳酸钾(287mg,2.08mmol)，室温搅拌2小时后，TLC监测反应结束。后处理：反应液倒入饱和食盐水中，乙酸乙酯萃取，有机相干燥并旋干，柱层析(乙酸乙酯：石油醚＝1：1)，得到白色泡沫状化合物60(560mg，两步收率90％)。
化合物60(100mg,0.064mmol)溶于0.8mL吡啶和3.2mL硫代乙酸中，60℃搅拌过夜，TLC监测反应完全。后处理：反应液加入硅胶旋干拌样，柱层析(二氯甲烷：甲醇＝40：1)，得到白色泡沫状化合物61(66.2mg，产率64.9％)。
化合物61(50mg,0.06mmol)溶于0.5mL 80％AcOH中，升温至70℃反应5小时，TLC监测反应完全；后处理：将反应液蒸干，柱层析(石油醚：乙酸乙酯＝1：3至二氯甲烷：甲醇＝30：1)，蒸干得白色泡沫状固体62(37mg，收率81.91％)。
HRMS(ESI)for C41H45NO17 823.2687,[M+H]+824.2714.
化合物62(37mg,0.05mmol)溶于1.5ml DMF中，向反应液中加入SO3·TMA(192.38mg,1.39mmol)，加毕升温至50℃反应10小时，TLC监测反应完全；后处理：反应液冷却后加入甲醇淬灭，搅拌0.5小时后蒸干反应液，柱层析(Sephadex LH-20，二氯甲烷：甲醇＝1：2)，快速硅胶柱层析(二氯甲烷：甲醇＝30：1至10：1)，蒸干得白色泡沫状化合物63(30mg，收率67.95％)。
HRMS(+ESI)for C41H45NO23S2 983.1824,[M+H]+984.1845,HRMS(-ESI)[M-H]-982.1832,[M/2-H]-490.5898.
化合物63(30mg,0.03mmol)溶于0.3ml Py中，将NH2NH2·H2O(16.93μL,0.30mmol)溶于0.3ml Py/AcOH中，滴加至反应液，室温反应0.5小时，TLC监测反应完全；后处理：向反应液中滴加丙酮(0.3mL)淬灭，搅拌0.5小时后蒸干反应液，得白色泡沫状固体，柱层析(Sephadex LH-20，二氯甲烷：甲醇＝1：2)，蒸干得白色泡沫状化合物64(18mg)，不经纯化直接投下一步。
HR(ESI)MS for C21H26NNa3O19S2 729.0234,[M+H]+730.0300.[M+Na]+752.0128.
化合物64(18mg,0.02mmol)溶于0.3ml MeOH中，向反应液中加入NaOH aq.(4N,0.3mL)，室温反应过夜，TLC监测反应完全；后处理：向反应液中加入Amberlite IR 120Hform ion-exchange resin中和至pH＝7，搅拌0.5小时后滤除树脂，蒸干滤液，柱层析(Sephadex G-25,100％H2O)，冻干得白色粉末状化合物34(11mg，两步收率49.42％)。
1H NMR(D2O,600MHz):δ7.12(d,J＝9.00Hz,2H,Ar-H of OMP),6.99(d,J＝9.00Hz,2H,Ar-H of OMP),5.03(d,J＝7.86Hz,1H,H-1),4.76-4.74(m,1H,H-4'),4.62(d,J＝7.80Hz,1H,H-1'),4.34(dd,J1＝11.28Hz,J2＝3.24Hz,1H,H-6'a),4.28(t,J＝8.70Hz,1H,H-6'b),4.14(dd,J1＝8.58Hz,J2＝3.24,1H,H-5'),3.96-3.93(m,1H,H-2'),3.91(dd,J1＝11.28Hz,J2＝3.24Hz,1H,H-3’),3.86-3.85(m,2H,H-4,H-5),3.82(s,3H,OCH3of OMP),3.74(t,J＝8.75Hz,1H,H-3),3.64(t,J＝8.75Hz,1H,H-2),2.07(s,3H,CH3of AcNH-).13C NMR(D2O,150MHz)δ177.93(C＝O of AcNH),176.53(C＝O of COOCH3),157.61(C of OMP),153.78(C of OMP),121.05(C of OMP),117.93(C of OMP),104.54(C-1'),104.07(C-1),84.35,79.49,78.31,76.85,75.30,75.18,72.71,70.59,58.68(OCH3of OMP),55.30,25.39(CH3of AcNH).
HR(ESI)MS for C21H26NNa3O19S2 729.0234,[M+H]+730.0300,[M+Na]+752.0128.
实施例2
化合物31(376mg,0.38mmol)溶于1.4mL吡啶中，加入新鲜配置的吡啶/乙酸/水合肼(v/v/v＝6/4/0.5)混合溶液7.0mL，室温搅拌40分钟，TLC监测反应结束。后处理：反应液用乙酸乙酯稀释，倒入3％稀盐酸，水相用乙酸乙酯萃取三次，干燥有机相并旋干，柱层析(二氯甲烷：丙酮＝50：1)，得到无色浆状化合物65(301mg，收率88.9％)。
1H NMR(CDCl3,500MHz)δ7.96(m,4H,ArH-Bz),7.48(t,J＝7.3Hz,1H,ArH-Bz),7.42–7.14(m,12H,ArH-Bz,PMB,benzylidene),6.94(d,J＝8.8Hz,2H,ArH-PMP),6.88(d,J＝8.3Hz,2H,ArH-PMB),6.76(d,J＝8.9Hz,2H,ArH-PMP),5.75(t,J＝9.4H z,1H,H1-3),5.63(t,J＝8.8Hz,1H,H1-2),5.26(s,1H,CH-benzylidene),5.14(d,J＝7.8Hz,1H,H1-1),4.67(d,J＝11.6Hz,1H,CH2-PMB),4.49(d,J＝11.6Hz,1H,CH2-PMB),4.29(t,J＝9.4Hz,1H,H1-4),4.19(d,J＝8.0Hz,1H,H2-1),4.02(dd,J＝10.9,2.9Hz,1H,H2-6a),3.92(d,J＝11.0Hz,1H,H2-6b),3.89(d,J＝2.5Hz,1H,H2-4),3.85(d,J＝9.4Hz,1H,H1-5),3.80(s,3H,OCH3-PMB),3.73(s,3H,OCH3-PMP),3.59(brs,2H,H1-6a,b),3.44(t,J＝9.0Hz,1H,H2-2),3.31(dd,J＝10.0,3.3Hz,1H,H2-3),2.94(s,1H,H2-5)；13C NMR(CDCl3,125MHz)δ165.67(CO-Bz),165.36(CO-Bz),159.37(ArC-PMB),155.65(ArC-PMP),151.29(ArC-PMP),137.41(ArC-benzylidene),133.21(ArC-Bz),132.82(ArC-Bz),130.14,129.92,129.82,129.75,129.40,129.20,128.40,128.14,128.07,126.57,118.92(ArC-PMP),114.53(ArC-PMP),113.85(ArC-PMB),101.35(C2-1),101.07(C1-1),100.84(CH-benzylidene),75.49(C1-4),75.15(C1-5),74.17(C2-3),73.32(C1-3),73.13(CH2-PMB),72.06(C2-4),71.77(C1-2),68.10(C1-6),67.61(C2-6),66.38(C-5),64.24(C2-2),55.63(OCH3-PMP),55.34(OCH3-PMB).HRMS(ESI)m/z:890.3110[M+H]+,912.2917[M+Na]+.Calcd.for C48H48N3O14 890.3136,C48H48N3NaO14912.2956.
化合物57(208.9mg,0.20mmol)和化合物65(150mg,0.17mmol)用无水甲苯带水三次，然后将其溶于5mL无水二氯甲烷中，加入800mg新烘干的分子筛。氩气保护下室温搅拌30分钟，然后将反应瓶置于-60℃，搅拌15分钟后，加入TMSOTf(106μL,0.19M,0.020mmol)的二氯甲烷溶液，继续搅拌30分钟后，TLC监测反应结束。后处理：加入过量三乙胺淬灭反应，然后过滤分子筛并浓缩滤液，柱层析(乙酸乙酯：石油醚＝1：3至1：1.5)，得到白色浆状化合物32(283.2mg，收率95％)。
1H NMR(CDCl3,500MHz)δ7.93-7.90(m,8H,ArH-Bz),7.48(m,2H,ArH-Bz),7.46-7.20(m,24H,ArH-Bz,PMB,benzylidene),6.91(d,J＝8.8Hz,2H,ArH-PMP),6.89(d,J＝8.0Hz,2H,ArH-PMB),6.86(d,J＝8.0Hz,2H,ArH-PMB),6.73(d,J＝8.5Hz,2H,ArH-PMP),5.71-5.66(m,2H,H1,3-3),5.59(t,J＝9.5Hz,1H,H1-2),5.37(t,J＝9.0Hz,1H,H3-2),5.22(s,1H,CH-benzylidene),5.17(s,1H,CH-benzylidene),5.09(d,J＝8.0Hz,1H,H1-1),4.95(d,J＝7.0Hz,1H,H3-1),4.64(d,J＝12Hz,1H,CH2-PMB),4.57-4.54(m,2H,H4-3,CH2-PMB),4.44(dd,J＝12Hz,2Hz,2H,CH2-PMB),4.28-4.17(m,4H,H1,3-4,H2,4-1),4.08(s,1H,H4-4),4.02(s,1H,H2-4),3.90-3.72(m,16H,H4-2,H1-5,H3-5,H3-6a,b,H4-6a,b,OCH3-PMB,OCH3-PMP),3.60-3.49(m,4H,H2-6a,b,H2-2,H1-6a),3.44(d,J＝12.0Hz,1H,H1-6b),3.21(d,J＝10.0Hz,1H,H2-3),2.85(s,1H,H2-5),2.80(s,1H,H4-5),2.70-2.69(m,2H,CH2-Lev),2.60-2.58(m,2H,CH2-Lev),2.06(s,3H,CH3-Lev)；13C NMR(125MHz,CDCl3)δ206.12(CO-Lev),171.99(CO-Lev),165.47(2C,CO-Bz),165.34(CO-Bz),165.10(CO-Bz),159.44(OMe-ArCof PMB),159.32(OMe-ArCof PMB),155.61(ArC-PMP),151.29(ArC-PMP),137.84(ArC-benzylidene),137.58(ArC-benzylidene),133.13(ArC-Bz),133.00(ArC-Bz),132.86(ArC-Bz),132.69(ArC-Bz),130.17(CH2-ArCof PMB),129.99(CH2-ArCof PMB),129.89(2C,ArC-Bz),129.81(8C,ArC-Bz),129.74(2C,ArC-Bz),129.73(CO-ArCof Bz),129.63(CO-ArCof Bz),129.57(4C,ArC-Bz),129.53(CO-ArCof Bz),129.42(CO-ArCof Bz),128.94,128.68,128.35,128.27,128.17,128.08,127.96,127.88,126.50(2C,ArC-PMB),126.48(2C,ArC-PMB),118.88(2C,ArC-PMP),114.49(2C,ArC-PMP),113.90(2C,ArC-PMB),113.88(2C,ArC-PMB),101.76(C4-1),101.73(C1-1),101.56(C3-1),100.71(C2-1),100.68(CH-benzylidene),100.48(CH-benzylidene),79.29(C2-3),76.18,75.46(C1,3-4),75.07,74.55(C1,3-5),74.50(C4-4),73.72(C1,3-3),73.36(CH2-PMB),73.32(C1,3-3),73.16(CH2-PMB),72.63(C4-3),72.23,72.16(C1,3-2),72.11(C2-4),68.55,67.99,67.51,66.44,66.14(C2,4-5),61.98,60.72(C2,4-2),55.62(OCH3-PMP),55.44(OCH3-PMB),55.33(OCH3-PMB),37.82(CH2-Lev),29.66(CH3-Lev),28.09(CH2-Lev).HRMS(ESI)m/z:1753.6017[M+H]+,1775.5819[M+Na]+.Calcd.for C48H48N3O14 1753.6038,C48H48N3NaO14 1775.5857.
化合物32(73mg,0.04mmol)溶于2mL二氯甲烷中，搅拌下加入水106μL，DDQ(23mg,0.1mmol)。室温剧烈搅拌过夜，TLC监测反应完全。后处理：反应液用二氯甲烷稀释，饱和碳酸氢钠水溶液洗涤两次，饱和食盐水洗涤一次，有机相干燥并旋干，柱层析(二氯甲烷：甲醇＝10：1)，得到白色固体化合物66(63.5mg，收率100％)。
1H NMR(CDCl3,500MHz)δ7.94-7.92(m,8H,ArH-Bz),7.48(m,2H,ArH-Bz),7.41-7.20(m,24H,ArH-Bz,benzylidene,overlapped with solvent peak),6.88(d,J＝8.5Hz,2H,ArH-PMP),6.75(d,J＝8.5Hz,2H,ArH-PMP),5.75(t,J＝9.5Hz,1H,H1-3),5.68(t,J＝8.5Hz,1H,H3-3),5.56(t,J＝8.5Hz,1H,H1-2),5.36(t,J＝7.5Hz,1H,H3-2),5.27(s,1H,CH-benzylidene),5.23(s,1H,CH-benzylidene),5.18(d,J＝8.0Hz,1H,H1-1),5.10(d,J＝6.5Hz,1H,H3-1),4.63(dd,J＝10.5,3.0Hz,1H,H4-3),4.38(d,J＝8.0Hz,1H,H2-1),4.32-4.28(m,3H,H1,3-4,H4-1),4.12(s,1H,H4-4),4.05-3.97(m,5H,H2-4,H1,3-6a,b),3.80(t,J＝9.0Hz,1H,H1-5),3.72-3.67(m,6H,H3-5,OCH3-PMP,H2-6a,b),3.61(br-s,2H,H4-6a,b),3.56-3.49(m,3H,H2-3,H2,4-2),3.06(s,1H,H2-5),2.93(s,1H,H4-5),2.71-2.69(m,2H,CH2-Lev),2.59-2.57(m,2H,CH2-Lev),2.06(s,3H,CH3-Lev)；13C NMR(125MHz,CDCl3)δ206.17(CO-Lev),172.03(CO-Lev),165.55(CO-Bz),165.41(2C,CO-Bz),165.18(CO-Bz),155.80(ArC-PMP),151.12(ArC-PMP),137.82(ArC-benzylidene),137.62(ArC-benzylidene),133.31(ArC-Bz),133.24(ArC-Bz),133.04(ArC-Bz),132.90(ArC-Bz),129.90,129.86,129.71,129.68,129.38,129.36,129.02,128.98,128.47,128.44,128.33,128.24,128.05,126.55,118.70(2C,ArC-PMP),114.72(2C,ArC-PMP),102.17(C4-1),101.93(C1-1),101.13(C3-1),100.76(C2-1),100.73(CH-benzylidene),100.66(CH-benzylidene),78.72(C2-3),75.82,75.53(C1,3-4),75.27,75.12(C1,3-5),74.61(C4-4),73.71(C1,3-3),73.21(C1,3-3),72.81(C4-3),72.33,72.14(C1,3-2),68.19(C2-4),68.07,66.58,66.34,62.29(C1,4-6),61.16,60.83(C2,4-5),60.71(C2,4-2),55.73(OCH3-PMP),53.56(C2,4-2),37.90(CH2-Lev),29.74(CH3-Lev),28.15(CH2-Lev).HRMS(ESI)m/z:1513.4835[M+H]+,1535.4673[M+Na]+.Calcd.for C78H77N6O26 1513.4888,C78H76N6NaO26 1535.4707.
化合物66(400mg，0.23mmol)，不经过柱层析，直接用于下步氧化，粗产品溶于12mL二氯甲烷和6mL NaHCO3-Na2CO3(PH 9.5)缓冲液中，室温剧烈搅拌，分多次加入TEPMPO(93.5mg，0.60mmol)和碘苯二乙酸(1.06g,3.28mmol)，室温搅拌48小时后，TLC监测反应结束。后处理：反应液倒入饱和食盐水中，二氯甲烷萃取，有机相干燥和旋干，得到黄色浆状化合物67，直接用于下一步反应。
以上粗产品溶解于3mL DMF中，加入碘甲烷(114μL，1.82mmol)和碳酸钾(94.5mg，0.68mmol)，室温搅拌3小时后，TLC监测反应结束。后处理：反应液倒入饱和食盐水中，乙酸乙酯萃取，有机相干燥并旋干，柱层析(二氯甲烷：丙酮＝50：1至20：1)，得到白色固体化合物68(200mg，三步收率56％)。
1H NMR(CDCl3,600MHz)δ7.97-7.93(m,5H,ArH-Bz),7.90(d,J＝7.2Hz,2H,ArH-Bz),7.52-7.24(m,23H,ArH-Bz,benzylidene),6.91(d,J＝9.0Hz,2H,ArH-PMP),6.77(d,J＝9.6Hz,2H,ArH-PMP),5.77(t,J＝9.0Hz,1H,H1-3),5.65-5.60(m,2H,H3-3,H4-3),5.34(t,J＝6.6Hz,1H,H3-2),5.38(s,1H,CH-benzylidene),5.21(d,J＝6.6Hz,1H,H1,3-1),5.21(s,1H,CH-benzylidene),5.12(d,J＝6.0Hz,1H,H1,3-1),4.55(dd,J＝9.6,8.4Hz,1H,H1-2),4.51-4.48(m,2H,H1,3-4),4.36(d,J＝7.8Hz,1H,H4-1),4.31-4.29(m,2H,H1,3-5),4.25(d,J＝3.0Hz,1H,H4-4),4.25(d,J＝8.4Hz,1H,H2-1),3.99(d,J＝3.0Hz,1H,H2-4),3.82(s,3H,OCH3-COOMe),3.75(s,3H,OCH3-COOMe),3.74(s,3H,OCH3-COOMe),3.72-3.69(m,1H,H4-2),3.66-3.61(m,2H,H2-2,H-6a),3.43-3.40(m,3H,H2-3,2H-6a,b),3.34(d,J＝9.0Hz,1H,H-6b),3.11(s,1H,H4-5),2.72-2.69(m,3H,CH2-Lev,H2-5),2.61-2.58(m,2H,CH2-Lev),2.07(s,3H,CH3-Lev)；13C NMR(CDCl3,150MHz)δ206.18(CO-Lev),172.07(CO-Lev),168.70(CO-COOMe),168.26(CO-COOMe),165.43(CO-Bz),165.32(CO-Bz),165.17(CO-Bz),164.92(CO-Bz),155.89(ArC-PMP),151.01(ArC-PMP),138.26(ArC-benzylidene),137.70(ArC-benzylidene),133.38(ArC-Bz),133.18(ArC-Bz),133.02(ArC-Bz),130.09,129.94,129.63,129.58,129.23,129.05,128.87,128.51,128.38,128.23,128.10,128.06,126.70,126.55,118.81(ArC-PMP),114.66(ArC-PMP),102.71(H4-1),102.46(H2-1),101.02(H1,3-1),100.97(H1,3-1),100.73(PhCH),100.58(PhCH),78.87(C2-3),77.96(C1-2),77.78(C1,3-4),74.47(C1,3-5),74.13(C1,3-5),73.54(C4-4),73.39(C3-3),72.37(C1,3-4),72.22(C2-3,C2-4),71.92(C3-2),71.80(C4-3),68.03(C4-6),67.92(C2-6),66.68(C4-5),66.19(C2-5),62.11(C2-2),60.62(C4-2),55.75(CH3-PMP),53.06(CH3-COOMe),52.97(CH3-COOMe),37.93(CH2-Lev),29.81(CH3-Lev),28.16(CH2-Lev).HRMS(ESI)m/z:1569.4762[M+H]+,1591.4579[M+Na]+.Calcd.for C80H77N3O28 1569.4786,C80H76N3NaO28 1591.4605.
方法1：化合物68(100mg,0.064mmol)溶于0.8mL吡啶和3.2mL硫代乙酸中，60℃搅拌8小时，补充1mL吡啶和1.5mL硫代乙酸，继续反应16小时，TLC监测反应完全。后处理：反应液加入硅胶旋干拌样，柱层析(二氯甲烷：甲醇＝40：1)，得到白色泡沫状化合物69(66.2mg，产率64.9％)。
方法2：化合物68(92.9mg,0.059mmol)溶于3.3mL吡啶和0.8mL水中，加入三乙胺(247μL，1.77mmol)和1,3-丙二硫醇(238μL，2.37mmol)，室温搅拌11小时，TLC监测反应完全。后处理：反应液旋干，溶于2mL甲醇和2mL二氯甲烷中，加入乙酸酐(200μL，0.24mmol)，室温搅拌1小时，TLC监测反应完全。后处理：柱层析(二氯甲烷：丙酮＝20：1至二氯甲烷：甲醇＝30：1)，得到白色泡沫状化合物69(58m g，收率61.2％)。
1H NMR(CDCl3,600MHz)δ8.05(d,J＝7.2Hz,2H,ArH-Bz),7.98-7.93(m,6H,ArH-Bz),7.59-7.25(m,22H,ArH-Bz,benzylidene),6.90(d,J＝9.0Hz,2H,ArH-PMP),6.75(d,J＝9.6Hz,2H,ArH-PMP),5.76(t,J＝9.0Hz,1H,H1-3),5.60(dd,J＝7.8,4.2Hz,1H,H3-3),5.57(s,1H,CH3-benzylidene),5.53(dd,J＝8.4,7.2Hz,1H,H1-2),5.50(d,J＝6.6Hz,1H,NH),5.25(s,1H,CH1-benzylidene),5.23(t,J＝4.2Hz,1H,H3-2),5.19(d,J＝9.0Hz,1H,H1-1),5.18(d,J＝6.6Hz,1H,H4-1),5.04(d,J＝4.2Hz,1H,H3-1),4.75(dd,J＝9.6,7.8Hz,1H,H3-4),4.70(dd,J＝10.8,3.6Hz,1H,H4-3),4.56-4.53(m,2H,H2-3,H1-4),4.42(d,J＝3.6Hz,1H,H4-4),4.39(d,J＝9.0Hz,1H,NH),4.34(d,J＝8.4Hz,1H,H4-1),4.30(d,J＝10.2Hz,1H,H1-5),4.23(d,J＝9.0Hz,1H,H3-5),4.02-3.98(m,1H,H4-2),4.31(d,J＝3.6Hz,1H,H2-4),3.80-3.72(m,11H,2H2,4-6a,b,OCH3-COOMe,OCH3-PMP),3.79(d,J＝11.4Hz,1H,H2,4-6a),4.49(d,J＝11.4Hz,1H,H2,4-6b),3.31-3.27(m,1H,H4-2),3.03(s,1H,H4-5),2.71-2.41(m,6H,H2-2,H2-5,CH2-Lev),2.04(s,3H,CH3-Lev),1.80(s,3H,CH3-NHAc),1.79(s,3H,CH3-NHAc)；13C NMR(CDCl3,150MHz)δ206.62(CO-Lev),172.22(CO-Lev),171.72(CO-COOMe),170.16(CO-COOMe),169.32(CO-NHAc),167.94(CO-NHAc),165.31(CO-Bz),165.25(CO-Bz),164.93(CO-Bz),164.87(CO-Bz),155.83(ArC-PMP),151.02(ArC-PMP),138.69(ArC-benzylidene),137.61(ArC-benzylidene),133.66(ArC-Bz),133.62(ArC-Bz),133.35(ArC-Bz),133.19(ArC-Bz),130.07,129.97,129.91,129.68,129.61,129.43,129.36,129.30,129.10,128.96,128.85,128.62,128.49,128.47,128.11,128.08,126.58,126.50,118.79(ArC-PMP),114.62(ArC-PMP),101.58(PhCH),100.78(PhCH),100.68(C1-1),99.75(C4-1),99.69(C3-1),98.99(C2-1),76.95(C3-4,overlapped with solventpeak),75.94(C1-4),75.04(C4-3,C3-3),74.66(C4-4),74.63(C4-4),73.13(C1-3),72.57(C2-4),72.53(C1-5,C3-2),72.03(C1-2),71.14(C2-3),68.77,68.27,66.72,66.22,55.74,55.05,53.08,52.83,49.88(H2-2),37.87,29.78,28.30,23.55,23.53.HRMS(ESI)m/z:1601.5145[M+H]+,1623.4974[M+Na]+.Calcd.for C84H85N2O30 1601.5187,C84H84N2NaO301623.5007.
化合物69(50mg,0.03mmol)溶于0.8mL 80％AcOH中，升温至70℃反应5小时，TLC监测反应完全；后处理：将反应液蒸干，柱层析(二氯甲烷：甲醇＝30：1至10：1)，蒸干得白色泡沫状固体70(38mg，收率53.37％)。
HR(ESI)MS for C70H76N2O30 1424.4483,[M+H]+1425.4498,[M+Na]+1447.4298.
化合物70(38mg,0.03mmol)溶于0.9mL DMF中，向反应液中加入SO3·TMA(150mg,1.08mmol)，加毕升温至50℃反应24小时，TLC监测反应完全；后处理：向反应液中加入甲醇淬灭，搅拌0.5小时后蒸干反应液，柱层析(Sephadex LH-20，二氯甲烷：甲醇＝1：2)，快速硅胶柱层析(二氯甲烷：甲醇＝30：1至10：1)，蒸干得白色泡沫状化合物71，68mg，不经进一步纯化直接投下一步。
HR(ESI)MS for C70H76N2O42S4 1744.2755[M+H]+1745.2754
化合物71(46mg,0.03mmol)溶于0.5mL Py中，将NH2NH2·H2O(14μL,0.25mmol)溶于0.5mLPy/AcOH中，滴加至反应液，室温反应0.5小时，TLC监测反应完全；后处理：向反应液中滴加0.5mL丙酮淬灭，搅拌0.5小时后蒸干反应液，得淡黄色油状物，柱层析(Sephadex LH-20，二氯甲烷：甲醇＝1：2)，蒸干得白色泡沫状固体72，50mg，不经纯化直接投下一步。HR(ESI)MS for C65H70N2O40S4,1646.2388[M+H]+1647.24622.
化合物72(50mg,0.03mmol)溶于0.3mL MeOH中，向反应液中加入NaOH aq.(4N,0.6mL)，室温反应过夜，TLC监测反应完全；后处理：向反应液中加入Amberlite IR 120Hform ion-exchange resin中和至pH＝7，搅拌0.5小时后滤除树脂，蒸干滤液，柱层析(Sephadex G-25,100％H2O)，冻干得白色粉末状化合物35(20mg，三步收率28.17％)。
1HNMR(400MHz,D2O)δ7.11(d,J＝9.08Hz,2H,Ar-H of OMP),6.98(d,J＝9.08Hz,2H,Ar-H of OMP),5.03(d,J＝7.84Hz,1H,H-11-GlcA),4.82-4.81(m,1H,H-42),4.74(d,J＝2.32Hz,1H,H-44),4.64(d,J＝7.76Hz,1H,H-12),4.60(d,J＝7.44Hz,1H,H-14),4.49(d,J＝7.80Hz,1H,H-13),4.34-4.30(m,2H,H-6a,H-6b),4.28-4.22(m,2H,H-6a,H-6b),4.14(dt,J1＝7.64Hz,J2＝3.32Hz,2H,H-52,H-54),4.09-4.07(m,2H,H-51,H-32),3.96-3.89(m,2H,H-22,H-53),3.88-3.84(m,2H,H-24,H-34),3.82(s,3H,CH3of OMP),3.78(t,J＝9.36Hz,1H,H-41),3.78-3.71(m,1H,H-43),3.69-3.63(m,2H,H-31,H-33),3.61(t,J＝9.04Hz,1H,H-21),3.40(t,J＝8.68Hz,1H,H-23),2.06(s,3H,CH3of AcNH),2.05(s,3H,CH3of AcNH).13C NMR(100MHz,D2O)δ177.93(C＝O of AcNH),177.91(C＝O of AcNH),177.10(C＝O of COOCH3),176.49(C＝O of COOCH3),157.60(C of OMP),153.78(C of OMP),121.04(C of OMP),117.92(C of OMP),106.59(C-13),104.67(C-12),104.35(C-14),104.04(C-11),84.88,84.64,79.51,79.47,78.94,78.31,78.15,76.89,76.63,75.27,75.10,74.97,72.75,70.71,70.41,58.67(CH3of OMP),55.27,54.25,25.43(CH3of AcNH),25.42(CH3of AcNH).
HR(ESI)MS for C35H44N2Na6O36S4 1333.9943,[M+H]+1335.0022,[M+Na]+1356.9852.
实施例3
化合物32(600mg，0.34mmol)溶于1.2mL吡啶中，加入新鲜配置的吡啶/乙酸/水合肼(v/v/v＝6/4/0.5)混合溶液6mL，室温搅拌40分钟，TLC监测反应结束。后处理：反应液用乙酸乙酯稀释，倒入3％稀盐酸，水相用乙酸乙酯萃取三次，干燥有机相并旋干，柱层析(二氯甲烷：丙酮＝25：1)，得到白色泡沫状化合物73(528.5mg，产率93.3％)。
1H NMR(CDCl3,500MHz)δ8.00-7.86(m,8H,ArH-Bz),7.46(t,J＝6.5Hz,2H,ArH-Bz),7.36-7.22(m,25H,ArH-Bz,PMB,benzylidene,overlapped with solvent peak),6.92-6.83(m,4H,ArH-PMP),6.83(d,J＝8.0Hz,2H,ArH-PMB),6.74(d,J＝8.0Hz,2H,ArH-PMB),6.73(d,J＝8.5Hz,2H,ArH-PMP),5.71-5.66(m,2H,H1,3-3),5.59(t,J＝8.0Hz,1H,H1-2),5.39(t,J＝7.5Hz,1H,H3-2),5.26(s,1H,CH-benzylidene),5.17(s,1H,CH-benzylidene),5.09(d,J＝8.0Hz,1H,H1-1),4.94(d,J＝7.0Hz,1H,H3-1),4.63(d,J＝11.5Hz,1H,CH2-PMB),4.58(d,J＝11.5Hz,1H,CH2-PMB),4.45-4.42(m,2H,CH2-PMB),4.26(t,J＝9.0Hz,1H,H1-4),4.17-4.08(m,5H,H3-4,H2,4-1,H2,4-4),3.94-3.72(m,17H,H4-3,H4-2,H1-5,H3-5,H3-6a,b,H4-6a,b,OCH3-PMB,OCH3-PMP),3.59-3.44(m,6H,H2-6a,b,H2-2,H1-6a),3.33(dd,J＝9.5Hz,1H,H1-6b),3.21(d,J＝10.0Hz,1H,H2-3),2.86(s,1H,H2-5),2.82(s,1H,H4-5)；13C NMR(125MHz,CDCl3)δ165.56(CO-Bz),165.52(CO-Bz),165.37(CO-Bz),165.14(CO-Bz),159.41(OMe-ArCof PMB),159.32(OMe-ArC of PMB),155.60(ArC-PMP),151.29(ArC-PMP),137.86(ArC-benzylidene),137.36(ArC-benzylidene),133.19(ArC-Bz),133.05(ArC-Bz),132.88,132.75,130.15,130.03,129.91,129.86,129.75,129.73,129.61,129.53,129.39,129.25,128.67,128.39,128.31,128.20,128.11,127.89,126.54,126.48,118.88(2C,ArC-PMP),114.50(2C,ArC-PMP),113.88(2C,ArC-PMB),101.86(C4-1),101.75(C2-1),101.56(C3-1),101.07(C1-1),100.72(CH-benzylidene),100.45(CH-benzylidene),79.34(C2-3),76.12,75.47(C1,3-4),75.05,74.60(C1,3-5),74.56,74.12(C2,4-4),73.65(C1,3-3),73.34(CH2-PMB),73.31(C1,3-3),73.17(CH2-PMB),72.08(C1,3-2),71.85(C4-3),68.50,68.07,67.49(C1,4-6),66.45,66.37(C2,4-5),64.38(C1,3-2),61.98,60.47(C2,4-2),55.64(OCH3-PMP),55.47(OCH3-PMB),55.37(OCH3-PMB).
化合物57(627.7mg,0.61mmol)和化合物73(845mg,0.51mmol)用无水甲苯带水三次，然后将其溶于10mL无水二氯甲烷中，加入2.5g新烘干的分子筛。氩气保护下室温搅拌30分钟，然后将反应瓶置于-60℃，搅拌15分钟后，加入TMSOTf(316μL,0.19M,0.061mmol)的二氯甲烷溶液，继续搅拌30分钟后，TLC监测反应结束。后处理：加入过量三乙胺淬灭反应，然后过滤分子筛并浓缩滤液，柱层析(二氯甲烷：丙酮＝60：1至20：1)，得到白色浆状化合物33(1.13g，收率88.1％)。
1H NMR(CDCl3,500MHz)δ7.94-7.86(m,12H,ArH-Bz),7.45-7.17(m,40H,ArH-Bz,PMB,benzylidene,overlapped with solvent peak),6.91-6.86(m,4H,ArH-PMP,PMB),6.82(m,4H,ArH-PMB),6.73(d,J＝8.5Hz,2H,ArH-PMP),5.71-5.56(m,4H,3H1,3,5-3,H1,3,5-2),5.38(t,J＝8.0Hz,1H,H1,3,5-2),5.34(t,J＝8.0Hz,1H,H1,3,5-2),5.22(s,1H,CH-benzylidene),5.17(s,1H,CH-benzylidene),5.15(s,1H,CH-benzylidene),5.09(d,J＝8.0Hz,1H,H1,3,5-1),4.95(d,J＝7.0Hz,1H,H1,3,5-1),4.89(d,J＝7.0Hz,1H,H1,3,5-1),4.62(d,J＝11.5Hz,1H,CH2-PMB),4.56-4.50(m,3H,CH2-PMB),4.43-4.41(m,2H,CH2-PMB,H6-3),4.36(d,J＝11.5Hz,1H,CH2-PMB),4.27-4.05(m,8H,3H1,3,5-4,3H2,4,6-1,2H2,4-4),4.02(s,1H,H6-4),3.89-3.73(m,19H,H6-2,3H1,3,5-5,6H-6a,b,OCH3-PMB),3.70(s,3H,OCH3-PMP),3.60-3.51(m,6H,4H-6a,b,H2,4-2),3.41(d,J＝12.0Hz,1H,H-6a),3.37(d,J＝11.5Hz,1H,H-6b),3.23(dd,J＝10.5Hz,2.5Hz,1H,H2,4-3),3.18(dd,J＝10.0Hz,2.0Hz,1H,H2,4-3),2.82(s,1H,H2,4,6-5),2.79(s,1H,H2,4,6-5),2.73(s,1H,H2,4,6-5),2.68-2.67(m,2H,CH2-Lev),2.59-2.57(m,2H,CH2-Lev),2.04(s,3H,CH3-Lev)；13C NMR(CDCl3,100MHz)δ206.16(CO-Lev),172.02(CO-Lev),165.49(CO-Bz),165.46(CO-Bz),165.36(CO-Bz),165.12(CO-Bz),165.11(CO-Bz),159.45(ArC-PMB),159.39(ArC-PMB),159.32(ArC-PMB),155.61(ArC-PMP),151.30(ArC-PMP),137.90(ArC-benzylidene),137.60(ArC-benzylidene),133.17(ArC-Bz),133.03(ArC-Bz),133.00(ArC-Bz),132.91(ArC-Bz),132.81(ArC-Bz),132.73(ArC-Bz),130.16,130.06,129.95,129.91,129.83,129.77,129.75,129.72,129.66,129.63,129.61,129.56,129.53,129.43,128.97,128.71,128.64,128.38,128.29,128.21,128.19,128.11,127.99,127.94,127.87,126.51,126.49,126.43,118.90(2C,ArC-PMP),114.50(2C,ArC-PMP),113.91(2C,ArC-PMB),113.89(4C,ArC-PMB),101.91(C1,3,5-1),101.73(C1,3,5-1),101.71(2C,C2,4,6-1),101.60(C2,4,6-1),100.73(C1,3,5-1),100.69(PhCH),100.42(2C,PhCH),79.39(C2,4-3),79.30(C2,4-3),76.23(C1,3,5-4),76.19(C1,3,5-4),75.49(C1,3,5-4),75.06(C1,3,5-5),74.53(C1,3,5-5),74.5 1(C1,3,5-5),74.46(2C,C1,3,5-3,C6-4),73.75(C1,3,5-3),73.71(CH2-PMB),73.39(CH2-PMB),73.35(CH2-PMB),73.17(2C,C1,3,5-3,C1,3,5-2),72.60(C6-3),72.22(C1,3,5-2),72.15(C1,3,5-2),72.14(C2,4-4),72.11(C2,4-4),68.58(2C,C-6),68.01(3C,C-6),67.52(C-6),66.46(2C,C2,4,6-5),66.16(C2,4,6-5),62.21(C2,4,6-2),61.97(C2,4,6-2),60.75(C2,4,6-2),55.65(OCH3-PMP),55.48(OCH3-PMB),55.44(OCH3-PMB),55.36(OCH3-PMB),37.84(CH2-Lev),29.71(CH3-Lev),28.11(CH2-Lev).HRMS(ESI)m/z:2519.8052[M+H]+,2540.8052[M+Na]+.Calcd.forC135H132N9O40 2518.8572,C135H131N9NaO40 2540.8391.
化合物33(1.274g,0.51mmol)溶于13mL二氯甲烷中，搅拌下加入水0.7mL，DDQ(413.4mg,1.82mmol)。室温剧烈搅拌5小时，TLC监测反应完全。后处理：反应液用二氯甲烷稀释，饱和碳酸氢钠水溶液洗涤两次，饱和食盐水洗涤一次，有机相干燥并旋干，柱层析(二氯甲烷：甲醇＝10：1)，得到白色泡沫状化合物74(1.15g，产率100％)。
1H NMR(CDCl3,500MHz)δ7.93-7.88(m,12H,ArH-Bz),7.48-7.21(m,33H,ArH-Bz,benzylidene),6.87(d,J＝8.0Hz,2H,ArH-PMP),6.75(d,J＝8.5Hz,2H,ArH-PMP),5.74(t,J＝9.5Hz,1H,H1,3,5-3),5.68-5.62(m,2H,H1,3,5-3),5.56(t,J＝8.5Hz,1H,H1,3,5-2),5.36-5.32(m,2H,H1,3,5-2),5.25(s,1H,CH-benzylidene),5.23(s,2H,CH-benzylidene),5.16(d,J＝7.5Hz,1H,H1,3,5-1),5.08-5.04(m,2H,H1,3,5-1),4.63(dd,J＝10.5,2.5Hz,1H,H6-3),4.38(d,J＝8.0Hz,1H,H2,4,6-1),4.30-4.23(m,5H,3H1,3,5-5or 4,2H2,4,6-1),4.12-3.90(m,8H,3H2,4,6-4,2H2,4,6-2,2H1,3,5-4or 5,H1,3,5-6a,b),3.79(t,J＝9.0Hz,1H,H1,3,5-4),3.74-3.43(m,16H,H2,4,6-2,2H2,4-3,OCH3-PMP,6H2,4,6-6a,b,4H1,3,5-6a,b),3.05(s,1H,H2,4,6-5),2.94(s,2H,H2,4,6-5),2.72-2.67(m,2H,CH2-Lev),2.59-2.57(m,2H,CH2-Lev),2.05(s,3H,CH3-Lev)；13C NMR(125MHz,CDCl3)δ206.27(CO-Lev),172.01(CO-Lev),165.58(CO-Bz),165.49(CO-Bz),165.44(2C,CO-Bz),165.28(CO-Bz),165.23(CO-Bz),155.79(ArC-PMP),151.09(ArC-PMP),137.73(ArC-benzylidene),137.59(ArC-benzylidene),133.32(ArC-Bz),133.24(2C,Ar C-Bz),133.04(ArC-Bz),132.90(2C,ArC-Bz),129.82,129.65,129.30,129.30,129.01,128.94,128.42,128.31,128.24,128.22,128.07,128.03,126.52,118.70(2C,ArC-PMP),114.71(2C,ArC-PMP),102.11(2C,C2,4,6-1),101.86(C2,4,6-1),101.16(C1,3,5-1),101.03(C1,3,5-1),100.69(4C,3C1,3,5-1,PhCH),78.59(C2,4-3),78.50(C2,4-3),75.71(C2,4,6-5or 4),75.50(C2,4,6-5or 4),75.39(C2,4,6-5or 4),75.17(C2,4,6-4),75.04(2C,C2,4,6-4or 5),74.53(C6-4),74.48(C2,4,6-3),73.70(C2,4,6-3),73.47(C2,4,6-3),73.15(C6-3),72.68(C2,4,6-2),72.24(2C,C2,4,6-2,C2,4-4),72.11(C2,4,6-2),68.06(2C,C2,4,6-6),68.00(C2,4,6-6),66.42(2C,C2,4,6-5),66.23(C2,4,6-5),62.20(2C,C2,4,6-2),61.02(C1,3,5-6),60.91(C1,3,5-6),60.69(C1/3/5-6,C2,4,6-2),55.70(OCH3-PMP),37.86(CH2-Lev),29.70(CH3-Lev),28.13(CH2-Lev).HRMS(ESI)m/z:2158.6836[M+H]+,2180.6694[M+Na]+.Calcd.for C111H108N9O37 2158.6846,C111H107N9NaO37 2180.6666.
化合物74(1.146mg,0.53mmol)溶于10mL乙腈和10mL NaHCO3-Na2CO3(PH 9.5)缓冲液中，室温剧烈搅拌，加入TEPMPO(100mg,0.64mmol)、碘苯二乙酸(1.28g,3.98mmol)、溴化钾(38mg,0.32mmol)、四丁基溴化铵(77mg,0.24mmol)，室温剧烈搅拌4小时后，TLC监测反应结束。后处理：反应用饱和硫代硫酸钠溶液淬灭，倒入饱和食盐水中，二氯甲烷萃取，有机相干燥和旋干，得到黄色浆状化合物75。
以上粗产品溶解于10mL DMF中，加入碘甲烷(756μL,12.14mmol)和碳酸钾(629.4mg,4.55mmol)，室温搅拌30分钟后，TLC监测反应结束。后处理：反应液倒入饱和食盐水中，乙酸乙酯萃取，有机相干燥并旋干，柱层析(乙酸乙酯：石油醚：二氯甲烷＝1：1：1)，得到白色固体化合物76(1.13g，两步收率46.2％)。
1H NMR(CDCl3,600MHz)δ7.97-7.83(m,11H,ArH-Bz),7.51-7.24(m,34H,ArH-Bz,benzylidene),6.91(d,J＝9.0Hz,2H,ArH-PMP),6.76(d,J＝9.0Hz,2H,ArH-PMP),5.76(t,J＝9.0Hz,1H,H1-3),5.65-5.60(m,3H,H3,5-3,H6-3),5.39-5.35(m,4H,H3,5-2,2H-CH-benzylidene),5.22(d,J＝6.6Hz,1H,H1,3-1),5.21(s,1H,CH-benzylidene),5.12(d,J＝6.0Hz,1H,H1,3-1),5.09(d,J＝6.0Hz,1H,H5-1),4.56(t,J＝9.0Hz,1H,H1-2),4.52-4.48(m,3H,H1,3,5-4),4.36(d,J＝7.8Hz,1H,H6-1),4.32(d,J＝10.2Hz,1H,H1,3-5),4.28(d,J＝9.0Hz,1H,H1,3-5),4.24(d,J＝9.6Hz,1H,H5-5),4.21(d,J＝2.4Hz,1H,H6-4),4.18(d,J＝3.0Hz,1H,H4-4),4.11(d,J＝8.4Hz,1H,H2,4-1),4.09(d,J＝8.4Hz,1H,H2,4-1),3.98(d,J＝3.0Hz,1H,H2-4),3.81(s,3H,OCH3-COOMe),3.73(s,3H,OCH3-COOMe),3.72(s,6H,OCH3-COOMe),3.69(dd,J＝10.8,6.4Hz,1H,H6-2),3.64-3.56(m,3H,H2,4-2,H-6a),3.56(d,J＝12.0Hz,1H,H-6b),3.42-3.39(m,3H,H2-3,2H-6a,b),3.35-3.34(m,2H,H-6a,b),3.30(dd,J＝10.2,3.0Hz,1H,H4-3),3.09(s,1H,H6-5),2.74(s,1H,H4-5),2.71-2.68(m,3H,CH2-Lev,H2-5),2.60-2.57(m,2H,CH2-Lev),2.06(s,3H,CH3-Lev)；13C NMR(CDCl3,150MHz)δ206.15(CO-Lev),172.01(CO-Lev),168.67(CO-COOMe),168.64(CO-COOMe),168.20(CO-COOMe),165.37(CO-Bz),165.27(CO-Bz),165.15(CO-Bz),165.11(CO-Bz),164.89(CO-Bz),155.85(ArC-PMP),150.95(ArC-PMP),138.22(ArC-benzylidene),138.19(ArC-benzylidene),137.65(ArC-benzylidene),133.37(ArC-Bz),133.30(ArC-Bz),133.16(ArC-Bz),133.07(ArC-Bz),132.99(ArC-Bz),130.03,129.89,129.87,129.59,129.52,129.18,129.01,128.83,128.70,128.47,128.44,128.35,128.31,128.20,128.06,128.04,127.95,126.60,126.49,118.76(ArC-PMP),114.62(ArC-PMP),102.67(C2,4-1),102.65(C2,4-1),102.43(C6-1),101.06(C5-1),100.91(C1,3-1),100.81(C1,3-1),100.67(PhCH),100.45(PhCH),100.40(PhCH),78.78(C2-3),78.74(C4-3),77.96(C1-3),77.93(C1,3,5-4),77.74(C1-2),74.41(C5-5),74.10(C2-4),73.98(C4-4),73.55(C1,3-5),73.46(C1,3-5),73.39(C3,5-3),73.27(C3,5-3),72.34(C1,3,5-4),72.31(C1,3,5-4),72.17(C6-4),71.89(C3,5-2),71.82(C3,5-2),71.75(C6-3),67.98(C-6),67.93(C-6),67.87(C-6),66.62(C6-5),66.45(C4-5),66.14(C2-5),62.17(C6-2),62.06(C4-2),60.58(C2-2),55.70(CH3-PMP),53.01(CH3-COOMe),52.93(CH3-COOMe),52.85(CH3-COOMe),37.87(CH2-Lev),29.75(CH3-Lev),28.10(CH2-Lev).HRMS(ESI)m/z:2242.6626[M+H]+,2264.6526[M+Na]+.Calcd.for C114H108N9O40 2242.6694,C114H107N9NaO40 2264.6513.
化合物76(114.9mg,0.051mmol)溶于1mL吡啶和2mL硫代乙酸中，60℃搅拌7小时，补充0.5mL吡啶和2mL硫代乙酸，继续反应17小时，TLC监测反应完全。后处理：反应液加入硅胶旋干拌样，柱层析(二氯甲烷：甲醇＝50：1)，得到白色泡沫状化合物77(61.6mg，产率52.5％)。
1H NMR(CDCl3,600MHz)δ8.06-7.92(m,12H,ArH-Bz),7.60-7.25(m,33H,ArH-Bz,benzylidene,overlapped with solvent peak),6.89(d,J＝9.0Hz,2H,ArH-PMP),6.75(d,J＝9.6Hz,2H,ArH-PMP),5.74(t,J＝9.0Hz,1H,H1,3,5-3),5.63(s,1H,CH-benzylidene),5.62-5.58(m,2H,H1,3,5-3,2),5.52(dd,J＝8.4,7.2Hz,1H,H1,3,5-2),5.44(s,1H,CH-benzylidene),5.40(d,1H,J＝6.6Hz,NH),5.25(s,1H,CH-benzylidene),5.23(t,J＝5.9Hz,1H,H1,3,5-3),5.18(d,J＝7.2Hz,1H,H1,3,5-1),5.17(d,J＝8.4Hz,1H,H1,3,5-1),5.09(t,1H,J＝3.3Hz,H1,3,5-2),5.00(d,J＝3.6Hz,1H,H1,3,5-1),4.99(d,J＝5.4Hz,1H,H2,4,6-1),4.81(d,J＝7.8,7.8Hz,1H,H1,3,5-4),4.78(br-d,J＝7.8Hz,1H,NH),4.70(d,J＝8.4Hz,1H,H2,4,6-1),4.66(dd,J＝11.4,3.6Hz,1H,H6-3),4.63(t,J＝8.4Hz,1H,H1,3,5-4),4.52(t,J＝9.0Hz,1H,H1,3,5-4),4.43(d,J＝3.6Hz,1H,H6-4),4.37(br-d,J＝7.2Hz,1H,H2,4-3),4.32-4.31(m,2H,H1,3,5-5,H2,4-4),4.22-4.19(m,3H,H2,4,6-1,2H1,3,5-5),4.04-4.02(m,2H,H2,4,6-2,H2,4-3),3.88(d,J＝3.6Hz,1H,H2,4-4),3.85(d,2H,J＝12.0Hz,H2,4,6-6a,b),3.78(d,J＝11.4Hz,1H,H2,4,6-6a),3.73-3.64(m,13H,OCH3-PMP,COOCH3,H2,4,6-6b),3.60-3.56(m,1H,H2,4,6-2),3.51(d,J＝11.4Hz,2H,H2,4,6-6a,b),3.26-3.22(m,1H,H2,4,6-2),2.99(s,1H,H2,4,6-5),2.80(s,1H,H2,4,6-5),2.70-2.39(m,5H,H2,4,6-5,CH2-Lev),2.03(s,3H,CH3-Lev),1.75(s,6H,CH3-NHAc),1.65(s,3H,CH3-NHAc)；13CNMR(CDCl3,125MHz)δ206.51(CO-Lev),172.16(CO-Lev),171.70,170.89,169.97,169.61,168.77,167.91,165.30(CO-Bz),165.21(CO-Bz),164.98(3C,CO-Bz),164.91(CO-Bz),155.83(ArC-PMP),151.03(ArC-PMP),138.64(ArC-benzylidene),138.58(ArC-benzylidene),137.59(ArC-benzylidene),133.68(2C,ArC-Bz),133.52(ArC-Bz),133.44(ArC-Bz),133.35(ArC-Bz),133.20(ArC-Bz),130.11,129.96,129.92,129.76,129.69,129.56,129.41,129.30,129.25,129.09,128.96,128.90,128.80,128.70,128.65,128.60,128.49,128.45,128.17,128.13,128.09,126.65,126.56,126.46,118.79(2C,ArC-PMP),114.62(2C,ArC-PMP),101.78(C1,3,5-1),100.81(C2,4,6-1),100.64(PhCH),100.32(C2,4,6-1),99.96(C2,4,6-1),99.87(PhCH),99.44(PhCH),99.40(C1,3,5-1),98.87(C1,3,5-1),76.41(C1,3,5-4),76.32(C1,3,5-4),75.85(C2,4-3),75.17(C1,3,5-4),75.06(C6-3),74.91(C1,3,5-3),74.67(C6-4),74.46(C1,3,5-2),74.24(C1,3,5-5),73.30(C2,4-4),73.05(C1,3,5-5),72.85(C1,3,5-3),72.48(C1,3,5-2),72.42(C2,4-4),72.19(C1,3,5-3),72.01(2C,C1,3,5-5,C1,3,5-2),71.25(C2,4-3),68.76(C2,4,6-6),68.67(C2,4,6-6),68.31(C2,4,6-6),66.66(C2,4,6-5),66.64(C2,4,6-5),66.16(C2,4,6-5),55.75(OCH3-PMP),54.90(C2,4,6-2),53.25(CH3-COOMe),53.05(C2,4,6-2),52.86(CH3-COOMe),52.80(CH3-COOMe),49.41(C2,4,6-2),37.84(CH2-Lev),29.77(CH3-Lev),28.29(CH2-Lev),23.53(CH3-NHAc),23.47(CH3-NHAc),23.43(CH3-NHAc).HRMS(ESI)m/z:2312.7107[M+Na]+.Calcd.forC114H107N9NaO4022312.7115.
化合物77(120mg,0.05mmol)溶于1.5mL 80％AcOH中，升温至70℃反应5小时，TLC监测反应完全；后处理：将反应液蒸干，柱层析(二氯甲烷：甲醇＝20：1至10：1)，蒸干得白色泡沫状固体78(55mg，收率47.83％)。
HR(ESI)MS for C99H107N3O43 2026.6312,[M+H]+2027.6307,[M/2+H]+1014.3209.
化合物78(55mg,0.03mmol)溶于3mL DMF中，向反应液中加入SO3·TMA(188.84mg,1.36mmol)，加毕升温至50℃反应24小时，TLC监测反应完全；后处理：向反应液中加入甲醇淬灭，搅拌0.5小时后蒸干反应液，柱层析(Sephadex LH-20，二氯甲烷：甲醇＝1：2)，快速硅胶柱层析(二氯甲烷：甲醇＝30：1至10：1)，蒸干得白色泡沫状化合物79，60mg，不经进一步纯化直接投下一步。
HR(ESI)MS for C99H107N3O61S6,2506.3721,[M+H]+2507.3462.
化合物79(60mg,0.02mmol)溶于0.6mL Py中，将NH2NH2·H2O(13.70μL,0.24mmol)溶于0.6mLPy/AcOH中，滴加至反应液，室温反应0.5小时，TLC监测反应完全；后处理：向反应液中滴加0.5mL丙酮淬灭，搅拌0.5小时后蒸干反应液，得淡黄色油状物，柱层析(SephadexLH-20，二氯甲烷：甲醇＝1：2)，蒸干得黄色固体80，60mg，不经纯化直接投下一步。
HR(ESI)MS for C94H101N3O59S6 2408.3353,[M+H]+2409.3164.
化合物80(60mg,0.03mmol)溶于0.5mL MeOH中，向反应液中加入NaOH aq.(4N,1.0mL)，室温反应过夜，TLC监测反应完全；后处理：向反应液中加入Amberlite IR 120Hform ion-exchange resin中和至pH＝7，搅拌0.5小时后滤除树脂，蒸干滤液，柱层析(Sephadex G-25,100％H2O)，冻干得白色粉末状化合物36(16mg，三步收率38.03％)。1HNMR(D2O,500MHz)δ7.13(d,J＝8.50Hz,2H,Ar-H of OMP),7.00(d,J＝8.50Hz,2H,Ar-H ofOMP),5.04(d,J＝8.05Hz,1H,H-11),4.86-4.79(m,2H,H-42,H-44),4.71(s,1H,H-46),4.70-4.61(m,3H,H-12,H-14,H-16),4.52-4.50(m,2H,H-13,H-15),4.34-4.24(m,6H,H-6a2,H-6b2,H-6a4,H-6b4,6a6,H-6b6),4.12-4.08(m,7H,H-51,H-52,H-54,H-56,H-22,H-24,H-34or H-32),3.92-3.87(m,4H,H-53,H-55,H-31,H-41),3.83(s,3H,CH3of OMP),3.76-3.71(m,5H,H-34orH-32,H-43,H-45,H-26,H-36),3.67-3.62(m,3H,H-33,H-35,H-21),3.43-3.40(m,2H,H-23,H-25),2.07(s,3H,CH3of AcNH),2.06(s,6H,2×CH3of AcNH).
HR(ESI)MS for C49H62N3Na9O53S6 1938.9652,[M+H]+1339.9707,[M/2+Na]+1356.9852.
实施例4
化合物27(250mg，0.25mmol)溶于1mL吡啶中，加入新鲜配置的吡啶/乙酸/水合肼(v/v/v＝6/4/0.5)混合溶液5mL，室温搅拌1小时，TLC监测反应完全；后处理：反应液用乙酸乙酯稀释，倒入3％稀盐酸，水相用乙酸乙酯萃取三次，干燥有机相并旋干，柱层析(乙酸乙酯：石油醚＝1：2)，得到白色泡沫化合物81(181.9mg，收率80.7％)。
1H NMR(CDCl3,500MHz)δ7.96-7.92(m,4H,ArH-Bz),7.48(t,J＝7.0Hz,1H,ArH-Bz),7.40–7.15(m,14H,ArH-Bz,PMB,benzylidene,SPhMe),7.06(d,J＝7.0Hz,2H,ArH-SPhMe),6.76(d,J＝8.0Hz,2H,ArH-PMB),5.68(t,J＝9.5Hz,1H,H1-3),5.36(t,J＝10.0Hz,1H,H1-2),5.24(s,1H,CH-benzylidene),4.86(d,J＝10.0Hz,1H,H1-1),4.63(d,J＝11.5Hz,1H,CH2-PMB),4.48(d,J＝11.5Hz,1H,CH2-PMB),4.21-4.17(m,2H,H1-4,H2-1),4.00(dd,J＝9.0Hz,1H,H2-6a),3.92(d,J＝11.0Hz,1H,H2-6b),3.85(d,J＝1.5Hz,1H,H2-4),3.80-3.74(s,4H,OCH3-PMB,H1-5),3.56(brs,2H,H1-6a,b),3.40(t,J＝8.5Hz,1H,H2-2),3.30(dd,J＝10.0,3.0Hz,1H,H2-3),2.89(s,1H,H2-5),2.30(s,3H,CH3-SPhMe)；13C NMR(CDCl3,125MHz)δ165.70(CO-Bz),165.31(CO-Bz),159.37(ArC-PMB),138.38(ArC-SPhMe),137.41(ArC-benzylidene),133.58(ArC-Bz),133.20(ArC-Bz),132.72,130.34,129.77,129.70,129.64,129.50,129.18,128.43,128.38,128.34,128.09,128.05,126.57,126.47,113.85(ArC-PMB),101.30(C2-1),101.09(CH-benzylidene),86.36(C1-1),79.21,75.32,74.63,74.20,73.13,71.82,70.95,68.12,67.90,66.37,64.24,55.37(OCH3-PMB),21.23(CH3-SPhMe).HRMS(ESI)m/z:912.2815[M+Na]+.Calcd.for C48H47N3NaO12S 912.2778.
化合物57(123mg,0.12mmol)和化合物81(89mg,0.1mmol)用无水甲苯带水三次，然后将其溶于3mL无水二氯甲烷中，加入400mg新烘干的分子筛。氩气保护下室温搅拌30分钟，然后将反应瓶置于-60℃，搅拌15分钟后，加入TMSOTf(63μL，0.19M，0.012mmol)的二氯甲烷溶液，继续搅拌30分钟后，TLC监测反应完全；后处理：加入过量三乙胺淬灭反应，然后过滤分子筛并浓缩滤液，柱层析(乙酸乙酯：石油醚＝1：4至3：5)，得到白色固体28(146.1mg,83.3％)。
1H NMR(CDCl3,400MHz)δ7.96-7.85(m,8H,ArH-Bz),7.46(m,2H,ArH-Bz),7.38-7.12(m,26H,ArH-Bz,PMB,benzylidene,SPhMe),7.04(d,J＝8.0Hz,2H,ArH-PMB),6.91(d,J＝8.0Hz,2H,ArH-SPhMe),6.83(d,J＝8.5Hz,2H,ArH-PMB),5.66(t,J＝8.8Hz,1H,H1,3-3),5.62(t,J＝9.2Hz,1H,H1,3-3),5.38-5.27(m,2H,H1,3-2),5.21(s,1H,CH-benzylidene),5.15(s,1H,CH-benzylidene),4.93(d,J＝7.2Hz,1H,H3-1),4.82(d,J＝10.4Hz,1H,H1-1),4.61(d,J＝11.2Hz,1H,CH2-PMB),4.56(d,J＝10.8Hz,1H,CH2-PMB),4.54(dd,J＝10.0,2.8Hz,1H,H4-3),4.44(d,J＝11.6Hz,2H,CH2-PMB),4.28(dd,J＝8.4,8.0Hz,1H,H1,3-4),4.23-4.13(m,4H,H1,3-4,H2-1,H2,4-4),4.09(d,J＝6.8Hz,1H,H4-1),4.28(dd,J＝17.2,2.8Hz,2H,H-6a,b),3.91-3.73(m,10H,H1,3-5,2H-H-6a,b,6H-OCH3-PMB),3.67(d,J＝11.8Hz,1H,H-6a),3.62(t,J＝6.0Hz,1H,H4-2),3.57-3.51(m,2H,H-6a,b),3.47(t,J＝6.8Hz,1H,H2-2),3.42(d,J＝12.0Hz,1H,H-6b),3.20(dd,J＝10.4,2.8Hz,1H,H2-3),2.83(s,1H,H2-5),2.79(s,1H,H4-5),2.72-2.69(m,2H,CH2-Lev),2.61-2.58(m,2H,CH2-Lev),2.30(s,3H,CH3-SPhMe),2.06(s,3H,CH3-Lev)；13C NMR(100MHz,CDCl3)δ206.19(CO-Lev),172.07(CO-Lev),165.61(CO-Bz),165.53(CO-Bz),165.37(CO-Bz),165.16(CO-Bz),159.50(OMe-ArCofPMB),159.38(OMe-ArCof PMB),138.38(ArC-SPhMe),137.91(ArC-benzylidene),137.65(ArC-benzylidene),133.58(ArC-Bz),133.20(ArC-Bz),133.07(ArC-Bz),132.92(ArC-Bz),132.66,132.48,131.03,130.40,130.06,129.96,129.92,129.90,129.84,129.82,129.79,129.73,129.70,129.64,129.58,129.55,129.01,128.73,128.54,128.40,128.33,128.24,128.18,128.09,128.03,127.94,126.56,126.54,113.96,113.93,101.81,101.72,101.63,100.76,100.75,100.53,86.38(C1-1),79.37,79.17,76.25,75.32,74.62,74.55,73.74,73.43,73.22,72.69,72.27,72.19,71.90,71.27,71.03,68.61,68.06,66.48,66.20,63.80,62.03,60.78,55.52(OCH3-PMB),55.40(OCH3-PMB),37.90(CH2-Lev),29.75(CH3-Lev),28.16(CH2-Lev),21.27(CH3-SPhMe).ESI-HRMS m/z:1775.5681[M+Na]+.Calcd.for C94H92N6NaO26S 1775.5680.
实施例5
化合物28(146mg，0.14mmol)溶于0.5mL吡啶中，加入新鲜配置的吡啶/乙酸/水合肼(v/v/v＝6/4/0.5)混合溶液2.6mL，室温搅拌40分钟，TLC监测反应完全；后处理：反应液用乙酸乙酯稀释，倒入3％稀盐酸，水相用乙酸乙酯萃取三次，干燥有机相并旋干，柱层析(乙酸乙酯：石油醚＝1：4至1：2)，得到白色泡沫状化合物82(117.6mg，收率85.3％)。
1H NMR(CDCl3,500MHz)δ7.93-7.85(m,8H,ArH-Bz),7.49-7.44(m,2H,ArH-Bz),7.37-7.16(m,26H,ArH-Bz,PMB,benzylidene,SPhMe),7.04(d,J＝8.0Hz,2H,ArH-PMB),6.91(d,J＝8.0Hz,2H,ArH-SPhMe),6.82(d,J＝8.0Hz,2H,ArH-PMB),5.66(t,J＝9.0Hz,1H,H1,3-3),5.62(t,J＝9.5Hz,1H,H1,3-3),5.37(t,J＝8.0Hz,1H,H1,3-2),5.32(t,J＝9.5Hz,1H,H1,3-2),5.25(s,1H,CH-benzylidene),5.16(s,1H,CH-benzylidene),4.93(d,J＝7.0Hz,1H,H3-1),4.81(d,J＝10.0Hz,1H,H1-1),4.60(d,J＝11.5Hz,1H,CH2-PMB),4.56(d,J＝11.5Hz,1H,CH2-PMB),4.54(d,J＝11.5Hz,2H,CH2-PMB),4.17-4.10(m,4H,H1,3-4,H2,4-1),4.05(s,1H,H2-4),3.93(dd,J＝10.5,2.8Hz,1H,H4-3),3.86-3.81(m,8H,H1,3-5,6H-OCH3-PMB),3.76(m,3H,H4-4,2H-6a,b),3.67(d,J＝9.5Hz,1H,H-6a),3.56(d,J＝10.5Hz,1H,H-6b),3.54-3.51(m,3H,H2,4-2,H-6a),3.45-3.42(m,2H,H-6a,b),3.31(d,J＝9.0Hz,1H,H-6b),3.20(dd,J＝10.5,3.0Hz,1H,H2-3),2.83(s,1H,H2-5),2.81(s,1H,H4-5),2.30(s,3H,CH3-SPhMe)；13C NMR(CDCl3,125MHz)δ165.57(2C,CO-Bz),165.35(CO-Bz),165.16(CO-Bz),159.47(OMe-ArCof PMB),159.37(OMe-ArCof PMB),138.36(ArC-SPhMe),137.93(ArC-benzylidene),137.41(ArC-benzylidene),133.56(ArC-Bz),133.19(ArC-Bz),133.04(ArC-Bz),132.87(ArC-Bz),132.64,130.39,130.10,129.93,129.89,129.78,129.72,129.62,129.54,129.26,128.65,128.52,128.38,128.32,128.21,128.14,128.07,127.89,126.56,126.51,113.92,101.86,101.72,101.57,101.12,100.48,86.36(C1-1),79.38,79.16,76.16,75.31,74.62,74.16,73.70,73.38,73.21,72.16,71.91,71.04,68.57,68.11,68.08,67.84,66.48,66.42,64.45,62.04,55.50(OCH3-PMB),55.39(OCH3-PMB),53.54,21.25(CH3-SPhMe).ESI-HRMS m/z:1655.5574[M+Na]+,1677.5315[M+Na]+.Calcd.for C89H87N6O24S 1655.5492,C89H86N6NaO24S 1677.5312.
化合物57(101.2mg,0.099mmol)和化合物82(109mg,0.066mmol)用无水甲苯带水三次，然后将其溶于3mL无水二氯甲烷中，加入400mg新烘干的分子筛。氩气保护下室温搅拌30分钟，然后将反应瓶置于-60℃，搅拌15分钟后，加入TMSOTf(41μL,0.19M,0.008mmol)的二氯甲烷溶液，继续搅拌30分钟后，TLC监测反应完全；后处理：加入过量三乙胺淬灭反应，然后过滤分子筛并浓缩滤液，柱层析(乙酸乙酯：石油醚＝1：2至1：1)，得到白色固体29(127.7mg，收率77.0％)。
1H NMR(CDCl3,500MHz)δ7.93-7.86(m,12H,ArH-Bz),7.47-7.43(m,3H,ArH-Bz),7.37-7.16(m,41H,ArH-Bz,PMB,benzylidene,SPhMe),7.03(d,J＝7.0Hz,2H,ArH-PMB),6.90(d,J＝7.5Hz,2H,ArH-SPhMe),6.82(m,4H,ArH-PMB),5.67(t,J＝9.0Hz,1H,H1,3,5-3),5.63-5.60(m,2H,H1,3,5-3),5.38(t,J＝8.0Hz,1H,H1,3,5-2),5.32(m,2H,H1,3,5-3),5.22(s,1H,CH-benzylidene),5.17(s,1H,CH-benzylidene),5.14(s,1H,CH-benzylidene),4.95(d,J＝7.0Hz,1H,H3-1),4.88(d,J＝6.5Hz,1H,H5-1),4.81(d,J＝10.0Hz,1H,H1-1),4.60(d,J＝12.0Hz,1H,CH2-PMB),4.55-4.50(m,3H,CH2-PMB,H6-3),4.42(d,J＝11.5Hz,2H,CH2-PMB),4.35(d,J＝11.5Hz,1H,CH2-PMB),4.23-4.02(m,9H,H1,3,5-4,H2,4,6-1,H2,4,6-4),3.88-3.876(m,19H,H1,3,5-5,9H-OCH3-PMB,7H-6a,b),3.66(d,J＝9.5Hz,1H,H-6a),3.58(t,J＝9.0Hz,1H,H6-2),3.54-3.51(m,4H,H2,4-2,2H-6a,b),3.38(t,J＝12.0Hz,2H,H-6a,b),3.20(d,J＝10.5Hz,1H,H2-3),3.17(d,J＝10.5Hz,1H,H4-3),2.80(s,1H,H2-5),2.79(s,1H,H4-5),2.73(s,1H,H6-5),2.70-2.68(m,2H,CH2-Lev),2.60-2.59(m,2H,CH2-Lev),2.29(s,3H,CH3-SPhMe),2.05(s,3H,CH3-Lev)；13C NMR(CDCl3,125MHz)δ206.13(CO-Lev),172.03(CO-Lev),165.56(CO-Bz),165.49(CO-Bz),165.39(CO-Bz),165.33(CO-Bz),165.14(CO-Bz),159.49(OMe-ArCof PMB),159.42(OMe-ArCof PMB),159.35(OMe-ArCof PMB),138.34(ArC-benzylidene),137.90(ArC-benzylidene),137.63(ArC-benzylidene),133.55(ArC-Bz),133.17(ArC-Bz),133.03(ArC-Bz),133.00(ArC-Bz),132.92(ArC-Bz),132.81(ArC-Bz),132.63,130.36,130.09,129.98,129.92,129.86,129.83,129.75,129.67,129.62,129.57,128.98,128.72,128.64,128.51,128.37,128.29,128.22,128.20,128.06,128.00,127.95,127.87,126.52,126.46,113.92,101.92,101.72(2C),101.69,101.62,100.72,100.45(2C),86.34(C1-1),79.41,79.31,79.13,76.24,76.20,75.30,74.62,74.56,74.48,73.75,73.41,73.37,73.18,72.64,72.24,72.20,71.02,68.60,68.03,67.82,66.47,66.20,62.24,61.99,60.78,55.48(OCH3-PMB),55.44(OCH3-PMB),55.36(OCH3-PMB),37.86,29.43,28.13,22.77(CH3-SPhMe).HRMS(ESI)m/z:2540.8296[M+Na]+.Calcd.for C135H131N9NaO38S2540.8213.
实施例6
化合物29(111.6mg,0.044mmol)溶于0.3mL吡啶中，加入新鲜配置的吡啶/乙酸/水合肼(v/v/v＝6/4/0.5)混合溶液1.6mL，室温搅拌40分钟，TLC监测反应结束。后处理：反应液用乙酸乙酯稀释，倒入3％稀盐酸，水相用乙酸乙酯萃取三次，干燥有机相并旋干，柱层析(二氯甲烷：丙酮＝25：1)，得到白色泡沫状化合物83(89.2m g，产率83.1％)。
1H NMR(CDCl3,500MHz)δ7.94-7.84(m,12H,ArH-Bz),7.50-7.43(m,3H,ArH-Bz),7.40-7.16(m,41H,ArH-Bz,PMB,benzylidene,SPhMe),7.04(d,J＝8.0Hz,2H,ArH-PMB),6.90(d,J＝8.8Hz,2H,ArH-SPhMe),6.82-6.80(m,4H,ArH-PMB),5.67(t,J＝8.8Hz,1H,H1,3,5-3),5.64-5.59(m,2H,H1,3,5-3),5.38(dd,J＝9.2,7.6Hz,1H,H1,3,5-2),5.34-5.29(m,2H,H1,3,5-3),5.26(s,1H,CH-benzylidene),5.18(s,1H,CH-benzylidene),5.13(s,1H,CH-benzylidene),4.93(d,J＝7.2Hz,1H,H3-1),4.87(d,J＝7.2Hz,1H,H5-1),4.81(d,J＝10.0Hz,1H,H1-1),4.59(d,J＝11.6Hz,1H,CH2-PMB),4.42(d,J＝11.6Hz,1H,CH2-PMB),4.35(d,J＝11.6Hz,1H,CH2-PMB),4.42(d,J＝11.6Hz,2H,CH2-PMB),4.35(d,J＝11.5Hz,1H,CH2-PMB),4.18-4.09(m,6H,H2,4,6-1,H1,3,5-4),4.05(d,J＝3.2Hz,1H,H2,4,6-4),4.05(d,J＝3.2Hz,1H,H2,4,6-4),3.92(dd,J＝11.2,4.4Hz,1H,H6-3),3.87-3.76(m,18H,H1,3,5-5,9H-OCH3-PMB,6H-6a,b),3.66(d,J＝9.6Hz,1H,H-6a),3.60-3.50(m,6H,H2,4-2,2H-6a,b,H6-2,H2,4,6-4),3.46-3.37(m,3H,H5-5,H-6a,b),3.32(dd,J＝10.0,4.0Hz,1H,H-6b),3.22(d,J＝10.4,3.2Hz,1H,H2-3),3.17(d,J＝10.4,3.6Hz,1H,H4-3),2.82(s,1H,H2-5),2.80(s,1H,H4-5),2.74(s,1H,H6-5),2.30(s,3H,CH3-SPhMe)；13C NMR(CDCl3,125MHz)δ165.59,165.42,165.37,165.19,159.48,159.43,159.36,138.37,137.88,137.37,133.54,133.20,133.07,133.03,132.93,132.84,132.67,130.36,130.08,130.02,129.95,129.90,129.85,129.81,129.79,129.75,129.73,129.72,129.71,129.66,129.56,129.52,129.30,128.71,128.68,128.52,128.40,128.33,128.30,128.24,128.21,128.16,128.08,127.95,127.89,126.56,126.53,126.46,113.92,101.95,101.84,101.79,101.71,101.60,101.13,100.49,100.45,86.37,79.43,79.33,79.11,77.48,77.36,77.16,76.84,76.20,76.18,75.32,74.67,74.62,74.56,74.52,74.48,74.16,73.74,73.67,73.38,73.19,72.15,72.14,71.90,71.02,68.56,68.53,68.11,68.05,68.03,67.81,66.49,66.45,66.42,64.43,62.23,61.98,55.52,55.47,55.41,21.26.HRMS(ESI)m/z:2420.8023[M+H]+.Calcd.forC130H126N9O36S 2420.8026.
化合物57(56mg,0.055mmol)和化合物83(88mg,0.036mmol)用无水甲苯带水三次，然后将其溶于3mL无水二氯甲烷中，加入250mg新烘干的分子筛。氩气保护下室温搅拌30分钟，然后将反应瓶置于-60℃，搅拌15分钟后，加入TMSOTf(36μL，0.1M，3.6μmol)的二氯甲烷溶液，继续搅拌30分钟后，TLC监测反应完全；后处理：加入过量三乙胺淬灭反应，然后过滤分子筛并浓缩滤液，柱层析(乙酸乙酯：石油醚＝1：2至1.5：1)，得到白色固体30(90.1mg，收率75.5％)，。
1H NMR(CDCl3,400MHz)δ7.93-7.85(m,16H,ArH-Bz),7.49-7.43(m,4H,ArH-Bz),7.40-7.15(m,54H,ArH-Bz,PMB,benzylidene,SPhMe),7.03(d,J＝8.0Hz,2H,ArH-PMB),6.90(d,J＝8.8Hz,2H,ArH-SPhMe),6.82(m,6H,ArH-PMB),5.67(t,J＝8.8Hz,1H,H1,3,5,7-3),5.64-5.58(m,3H,H1,3,5,7-3),5.39-5.29(m,4H,H1,3,5-3),5.26(s,1H,CH-benzylidene),5.17(s,1H,CH-benzylidene),5.14(s,1H,CH-benzylidene),5.13(s,1H,CH-benzylidene),4.94(d,J＝7.2Hz,1H,H3-1),4.89(d,J＝6.4Hz,1H,H5-1),4.87(d,J＝6.8Hz,1H,H7-1),4.81(d,J＝10.0Hz,1H,H1-1),4.59(d,J＝11.6Hz,1H,CH2-PMB),4.55-4.52(m,2H,CH2-PMB,H8-3),4.49(d,J＝11.2Hz,2H,CH2-PMB),4.43(d,J＝11.6Hz,2H,CH2-PMB),4.33(d,J＝11.6Hz,2H,CH2-PMB),4.28-4.02(m,12H,H1,3,5,7-4,H2,4,6,8-1,H2,4,6,8-4),3.88-3.72(m,25H,H1,3,5,7-5,12H-OCH3-PMB,9H-6a,b),3.66(d,J＝10.0Hz,1H,H-6a),3.56-3.48(m,8H,H2,4,6,8-2,4H-6a,b),3.24-3.14(m,3H,H2,4,6-3),2.79(s,2H,H2,4-5),2.72(s,1H,H6-5),2.70-2.68(m,2H,CH2-Lev),2.61-2.57(m,3H,CH2-Lev,H8-5),2.27(s,3H,CH3-SPhMe),2.06(s,3H,CH3-Lev)；13C NMR(CDCl3,125MHz)δ206.17,172.05,165.57,165.50,165.38,165.35,165.16,165.14,159.49,159.44,159.42,159.36,138.35,137.93,137.90,137.63,133.55,133.19,133.05,133.01,132.94,132.83,132.64,130.37,130.09,130.05,129.99,129.94,129.89,129.87,129.84,129.83,129.81,129.79,129.76,129.74,129.71,129.70,129.68,129.63,129.59,129.58,129.54,129.00,128.74,128.67,128.66,128.64,128.53,128.39,128.31,128.29,128.24,128.21,128.07,128.02,127.97,127.92,127.88,101.95,101.94,101.73,101.70,101.64,100.73,100.45,100.40,86.36,79.42,79.39,79.32,79.13,77.36,76.26,76.24,75.31,74.63,74.62,74.57,74.54,74.51,74.50,74.48,74.45,73.79,73.78,73.77,73.74,73.42,73.38,73.36,73.19,72.65,72.25,72.20,72.18,72.17,71.02,69.60,68.59,68.05,68.03,67.81,66.49,66.48,66.47,66.20,62.24,62.20,61.98,60.78,55.50,55.46,55.38,53.88,37.87,29.45,28.14,22.79.HRMS(MALDI/CHCA)m/z:3306.9292[M+Na]+,3322.8774[M+K]+.Calcd.forC176H170N12NaO50S 3306.0747,C176H170N12O50S 3322.0487.
实施例7
化合物57(1.69g,1.65mmol)和2-benzyloxycarbonylaminoethanol(0.58g,2.95mmol)混合，甲苯带水(5mL×3)，蒸干后溶解于17mL二氯甲烷中，加入5g新烘干的分子筛，室温搅拌0.5小时，降温至0℃，向反应液中滴加TMSOTf(1M,302μL)，继续搅拌0.5h，TLC监测反应完全；滴加三乙胺淬灭，过滤蒸干，柱层析(石油醚:乙酸乙酯＝2：1)，蒸干得白色泡沫状固体84(1.28g，收率74.22％)。HRMS for 1058.3797[M+H]+1059.3863
化合物84(1.0g,0.94mmol)溶于17mL二氯甲烷中，避光，向反应液中加入DDQ(0.26g,1.13mmol)，室温搅拌3小时，TLC监测反应完全；加入饱和碳酸氢钠溶液淬灭，二氯甲烷萃取(10mL×3)，合并有机相，饱和氯化钠溶液洗涤,有机相用无水硫酸钠干燥，过滤，蒸干，柱层析(石油醚：乙酸乙酯＝1：1)，蒸干得白色泡沫状固体85(0.64g，收率72.23％)。HRMS for 938.3222[M+H]+939.3261
化合物85(0.64g,0.68mmol)溶于7mL乙腈和碱性溶液(pH＝9.5,7mL)中，剧烈搅拌，向反应液中加入KBr(16.23mg,0.14mmol)，TEMPO(42.63mg,0.27mmol)和BAIB(0.55g,1.71mmol))，室温搅拌3小时，TLC监测反应完全；向反应液中滴加Na2S2O3稀溶液淬灭，搅拌0.5h后二氯甲烷萃取(10mL×3)，合并有机相，饱和氯化钠溶液洗涤，有机相用无水硫酸钠干燥，过滤，蒸干得黄白色泡沫状固体86，0.65g，不经纯化直接投下一步。
化合物86(0.65g,0.68mmol)溶于7mL DMF中，向反应液中加入K2CO3(0.28g,2.05mmol)，CH3I(0.34mL,5.46mmol)，室温搅拌5小时，TLC监测反应完全；将反应液倒入水中，二氯甲烷萃取(10mL×3)，合并有机相，饱和氯化钠溶液洗涤,有机相用无水硫酸钠干燥，过滤，蒸干，柱层析(石油醚：乙酸乙酯＝1：1)，蒸干得白色泡沫状固体87(0.52g，两步收率78.91％)。HRMS for 966.3171[M+H]+967.3253
化合物87(0.5g,0.52mmol)溶于7mL吡啶中，向反应液中加入AcSH(14mL)，，室温搅拌15小时，TLC监测反应完全；将反应液浓缩为黄色油状物，柱层析(乙酸乙酯：甲醇＝10：1)，蒸干得白色泡沫状固体88(0.51g，收率98.43％)。HRMS for 982.3372[M+H]+983.3414
化合物88(0.5g,0.51mmol)溶于5mL 80％AcOH中，70℃搅拌5小时，TLC监测反应完全；将反应液蒸干，柱层析(二氯甲烷：甲醇＝30：1)，蒸干得白色泡沫状固体89(0.38g，收率83.48％)。HRMS for 894.3059[M+H]+895.3064
化合物89(150mg,0.17mmol)溶于8mL DMF中，向反应液中加入SO3.TMA(700mg,5.03mmol)，50℃搅拌22小时，TLC监测反应完全；向反应液中加入甲醇淬灭，蒸干反应液，柱层析(Sephadex LH-20，二氯甲烷：甲醇＝1：2)，蒸干得白色泡沫状固体90(130mg，收率73.52％)。HRMS for 1055.2258[M+H]+1055.2258[M+Na]+1077.2070
化合物90(130mg,0.12mmol)溶于1mL Py中，将NH2NH2·H2O(68.39μL,1.20mmol)溶于Py/AcOH(1mL)中，加入反应液，室温反应30分钟，TLC监测反应完全；蒸干反应液，得淡黄色油状物，柱层析(Sephadex LH-20，二氯甲烷：甲醇＝1：2)，蒸干得白色泡沫状固体91(80mg，收率81.09％)。HRMS for 956.1827[M+H]+957.1909
化合物91(80mg,0.10mmol)溶于0.8mL MeOH中，向反应液中加入NaOH溶液(4N,0.8mL)，室温搅拌过夜，TLC监测反应完全；向反应液中加入Amberlite IR 120H form ion-exchange resin中和至Ph中性，滤除树脂，蒸干滤液，柱层析(Sephadex G-25,100％H2O)，冻干得白色粉末40(24mg，两步收率24.49％)。
1H NMR(400MHz,D2O)δ7.47-7.40(m,5H,Ar-H of Cbz),5.12(s,2H,CH2of Cbz),4.73(s,1H,H-42),4.60(d,J＝6.50Hz,1H,H-12),4.44(d,J＝7.72Hz,1H,1),4.32(dd,J1＝11.04Hz,J2＝2.12Hz,1H,H-32)，4.25(t,J＝8.60Hz,1H,H-51),4.12(dd,J1＝7.60Hz,J2＝1.96Hz,1H,H-41),3.92-3.88(m,3H,H-22,H-52,CH2of OCH2CH2NH-Cbz),3.75-3.70(m,3H,CH2ofOCH2CH2NHCbz,H-6a,H-6b),3.60(t,J＝8.92Hz,1H,H-31),3.37-3.33(m,2H,H-21,CH2of OCH2CH2NHCbz),2.89(s,1H,CH2of OCH2CH2NHCbz),2.05(s,3H,CH3of AcNH).
实施例8
化合物89(100mg,0.11mmol)溶于2mL DMF中，向反应液中加入SO3.TMA(46.68mg,0.33mmol)，50℃搅拌5小时，TLC监测反应完全；向反应液中加入甲醇淬灭，搅拌0.5h后蒸干反应液，柱层析(二氯甲烷：甲醇＝10：1)，蒸干得白色泡沫状固体92，100mg。HRMS for974.2627[M+H]+975.2706
化合物93(110mg,0.11mmol)溶于1mL Py中，将NH2NH2.H2O(62.54μL,1.09mmol)溶于Py/AcOH(1mL)中加入反应液，室温搅拌30分钟，TLC监测反应完全，蒸干反应液，柱层析(二氯甲烷：甲醇＝5：1)，蒸干得白色泡沫状固体93(50mg，两步收率62.21％)。HRMS for876.2259[M+H]+877.2325
化合物93(50mg,0.07mmol)溶于0.5mL MeOH中，向反应液中加入NaOH溶液(4N,0.5mL)，室温搅拌过夜，TLC监测反应完全；向反应液中加入Amberlite IR 120H form ion-exchange resin中和至Ph中性，滤除树脂，蒸干滤液，柱层析(Sephadex G-25,100％H2O)，冻干得白色粉末37(18mg，收率45.18％)。
1H NMR(D2O,400MHz)δ7.44-7.39(m,5H,Ar-H of Cbz),5.11(s,2H,CH2of Cbz),4.51(d,J＝8.32Hz,2H,H-12,H-11),4.23(d,J＝6.16Hz,2H,H-51,H-41),3.99(d,J＝2.72Hz,1H,H2),3.98-3.95(m,1H,H-31),3.91(d,J＝8.64Hz,1H,H-21),3.90-3.87(m,1H,H-22),3.82-3.73(m,2H,H-6a,H-6b),3.74-3.72(m,3H,H-32,CH2of OCH2CH2NHCbz),3.68-3.63(m,1H,H-52),3.37-3.32(m,2H,CH2of OCH2CH2NHCbz),2.02(s,3H,CH3of AcNH).13C NMRδ174.88(C＝O of AcNH),170.89(C＝O of COONa),158.47(C＝O of Cbz),136.59(Ar-C ofCbz),128.86,128.45,127.74,102.62(C-12),101.97(C-11),80.70,73.91,73.37,72.88,72.27,70.48,69.26,67.34,66.97,55.53,52.25,40.48(C of CH2NHCbz),22.42(CH3ofAcNH).
实施例9
化合物89(130mg,0.15mmol)溶于1.5mL吡啶中，向反应液中加入BzCN(38.12mg,0.29mmol)，室温搅拌16小时，TLC监测反应完全；向反应液中加入甲醇淬灭，蒸干反应液，柱层析(二氯甲烷：甲醇＝20：1)，蒸干得白色泡沫状固体94(80mg，收率55.17％)。HRMSfor998.3321[M+H]+999.3408.
化合物94(80mg,0.08mmol)溶于1mL DMF中，向反应液中加入SO3.TMA(111.52mg,0.80mmol)，50℃搅拌过夜，TLC监测反应完全；向反应液中加入甲醇淬灭，蒸干反应液，柱层析(二氯甲烷：甲醇＝20：1)，蒸干得白色泡沫状固体95(65mg，收率75.25％)。HRMS for974.2627[M+H]+975.2706
化合物95(65mg,0.06mmol)溶于0.5mL Py中，将NH2NH2·H2O(33.43μL,0.58mmol)溶于Py/AcOH(0.5mL)中，滴加至反应液，室温搅拌30分钟，TLC监测反应完全；蒸干反应液，得淡黄色油状物，柱层析(二氯甲烷：甲醇＝10：1)，蒸干得白色泡沫状固体96(35mg，收率59.32％)。HRMS for 980.2521[M+H]+981.2637
化合物96(30mg,0.03mmol)溶于0.5mL MeOH中，向反应液中加入NaOH溶液(4N,0.5mL)，室温搅拌过夜，TLC监测反应完全；向反应液中加入Amberlite IR 120H form ion-exchange resin中和至Ph至中性，滤除树脂，蒸干，柱层析(Sephadex G 25,100％H2O)，冻干得白色粉末38(12mg,收率56.18％)。
1H NMR(D2O,400MHz)δ7.47-7.39(m,5H,Ar-H of Cbz),5.12(s,2H,CH2of Cbz),4.68(s,1H,H-42),4.56-4.53(m,1H,H-12),4.49(d,J＝7.76Hz,1H,H-11),3.96-3.91(m,1H,H-51),3.89-3.87(m,2H,H-41,H-22),3.83-3.77(m,5H,CH2of OCH2CH2NHCbz,H-52,H-32,H-6a),3.75-3.73(m,1H,H-6b),3.68-3.63(m,2H,H-31,H-21),3.38-3.33(m,2H,CH2ofOCH2CH2NHCbz),2.04(s,3H,CH3of AcNH).13C NMRδ174.93(C＝O of AcNH),171.70(C＝O ofCOONa),158.50(C＝O of NHC＝O),136.59(Ar-C of Cbz),128.85,128.43,127.73,102.66(C-12),101.62(C-11),80.28,75.59,74.58,74.08,73.90,72.36,69.76,69.24,66.98,61.06,52.75,40.49(C of CH2NHCbz),22.43(CH3of AcNH).
实施例10
化合物89(80mg，0.20mmol)溶于2.0mL Py中，将NH2NH2·H2O(111.63μL,1.95mmol)溶于Py/AcOH(2.0mL)中，加入反应液，室温搅拌30分钟，TLC监测反应完全；蒸干反应液，得淡黄色油状物，柱层析(SephadexLH-20，二氯甲烷：甲醇＝1：2)，蒸干得淡黄色泡沫状固体97(148mg，收率92.34％)。HRMS for 796.2691[M+H]+797.2737
化合物97(50mg,0.06mmol)溶于0.5mL MeOH中，向反应液中加入NaOH溶液(4N,0.5mL)，室温搅拌过夜，TLC监测反应完全；向反应液中加入Amberlite IR 120H form ion-exchange resin中和至Ph至中性，滤除树脂，蒸干，柱层析(Sephadex G-10，100％H2O)，冻干得白色粉末39(20mg，收率53.42％)。HRMS for 597.1873[M+H]+797.2737[M-Na+2H]+575.2070.
1H NMR(D2O,400MHz)δ7.46-7.39(m,5H,Ar-H of Cbz),5.12(s,2H,CH2of Cbz),4.48(d,J＝8.52Hz,1H,H-12),4.43(m,1H,H-11),3.92-3.82(m,3H,H-51,H-41,H-22),3.79-3.65(m,8H,CH2of OCH2CH2NHCbz,H-42,H-32,H-6a,H-6b,H-31,H-21),3.35-3.31(m,3H,H-52,CH2ofOCH2CH2NHCbz),2.05(s,3H,CH3of AcNH).13C NMRδ175.13(C＝O of AcNH),174.34(C＝O of COONa),158.53(C＝O of Cbz),136.60(Ar-C of Cbz),128.85,128.43,127.73,102.63(C-12),101.09(C-11),79.65,76.63,75.29,73.89,72.74,71.15,69.06,67.85,66.98,61.12,52.39,40.48(C of CH2NHCbz),22.52(CH3of AcNH).
实验例部分
实验例1药理实验：与Midkine蛋白相互作用
本发明利用下述相互作用测试提供了硫酸软骨素寡糖与Midkine蛋白的相互作用亲和力。
(1)试验方法：
采用直接偶联法将Midkine、Pleiotrophin蛋白偶联于Fc2、Fc3通道上；结合缓冲液体系为HBS-EP+，pH7.4。将配制好的一系列浓度的多糖流经芯片表面，进行相互作用测定。
蛋白偶联：实验采用氨基偶联法，将Midkine、Pleiotrophin蛋白偶联于Fc2、Fc3通道上，Fc1通道作为参比通道；Midkine、Pleiotrophin偶联条件为：50μg/ml，pH5.0醋酸钠缓冲液；活化时间为420s；手动偶联模式进样；封闭时间为420s；为减少参比通道非特异性吸附，将参比通道进行了活化再封闭处理。
运行缓冲液：HBS-EP+
(2)多糖-蛋白相互作用测试
根据表面测试结果，将多糖配置成如下所示浓度：
(3)多糖与蛋白亲和力测定结果见下表
多糖与蛋白质亲和力测定结果表
Claims (7)
1.一类如通式(I)所示的一系列不同亚型的硫酸软骨素寡糖及其药学上可接受的盐，具有以下的结构通式：
n＝1,2,3，
R1选自OH或OSO3Na，R2选自OH或OSO3Na，R3选自对甲氧基苯基或苄氧羰基氨基-1-乙基。
2.根据权利要求1的硫酸软骨素寡糖及其药学上可接受的盐，其特征在于，
其中，n＝1,2,3。
3.根据权利要求1的硫酸软骨素寡糖及其药学上可接受的盐，其特征在于，
其中，R1选自OH或OSO3Na，R2选自OH或OSO3Na。
4.根据权利要求1的硫酸软骨素寡糖及其药学上可接受的盐，所述的化合物选自如下：
化合物34
化合物35
化合物36
化合物37
化合物38
化合物39
化合物40
5.权利要求1～4中任一硫酸软骨素寡糖的制备方法，其特征在于，包括如下步骤：
从葡萄糖和氨基半乳糖开始，经过一系列保护基策略，得到了适当保护的单糖受体和供体，然后应用溶剂效应，糖基化反应得到二糖重复单元；然后应用“先糖苷化-后氧化策略”，先延长糖链，然后经过氧化甲酯化和还原乙酰化得到全保护的硫酸软骨素寡糖；再经过脱苄叉、硫酸化和脱保护，可以得到相应的硫酸软骨素寡糖；
1)全保护硫酸软骨素寡糖中间体的制备
上述的方法，包括下列步骤：
(1).在葡萄糖的端基引入对甲苯硫醚保护、2,3-位引入苯甲酰基保护、6-位引入对甲氧基苯甲醚保护，得到了相应保护的单糖受体；
(2).在氨基半乳糖的端基引入三氯亚胺酯保护、2-位引入叠氮保护、3-位引入乙酰丙酸酯保护、4,6-位引入苯甲缩醛保护，得到了相应保护的单糖供体；
(3).将上述得到的单糖受体和供体在酸性条件下，进行糖基化反应，合成出二糖重复单元；
(4).将上述得到的二糖单元，在酸性条件下脱除端基的硫醚保护，并转化成三氯亚胺酯，得到二糖供体；
(5).将上述得到的二糖单元，在水合肼的作用下脱除4-位的乙酰丙酸保护，得到二糖受体；
(6).将上述得到的二糖供体和受体在酸性条件下，进行糖基化反应，延长糖链得到四糖，并用同样的方法延长糖链得到六糖；
2).全保护硫酸软骨素寡糖中间体端基的转化
上述的方法，包括以下步骤：
(1).将上述得到的全保护的硫酸软骨素寡糖在酸性条件下将端基的甲氧基苯基脱除得到中间体8；
(2).将上述得到的中间体8，在酸性条件下将端基转化为苄氧羰基氨基-1-乙基保护；
3).硫酸软骨素寡糖的制备
上述的方法，包括以下步骤：
(1).将葡萄糖单元6-位的对甲氧基苯甲醚保护用DDQ脱除，然后用氧化剂将伯醇氧化为羧酸，再甲酯化转化为羧酸甲酯；
(2).将氨基半乳糖单元2-位的叠氮，用还原剂还原成氨基，并乙酰化得到乙酰氨基，从而得到全保护的硫酸软骨素寡糖；
(3).将上述得到的全保护的硫酸软骨素寡糖，在酸性条件下将苯甲缩醛保护脱除，然后用硫酸化试剂在不同的条件下对4,6-位选择性硫酸化，最后分别在水合肼和碱性条件下将酯基保护脱除，得到了相应的硫酸软骨素寡糖。
6.权利要求1-4任一项的硫酸软骨素寡糖及其药学上可接受的盐在制备预防或治疗与肝素结合细胞因子Midkine相关疾病药物中的应用。
7.根据权利要求6的应用，其特征在于，所述的与肝素结合细胞因子Midkine相关疾病包括艾滋病、神经病变性疾病、肿瘤。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810033328.1A CN110041383A (zh) | 2018-01-15 | 2018-01-15 | 硫酸软骨素寡糖、其制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810033328.1A CN110041383A (zh) | 2018-01-15 | 2018-01-15 | 硫酸软骨素寡糖、其制备方法和应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110041383A true CN110041383A (zh) | 2019-07-23 |
Family
ID=67272671
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810033328.1A Pending CN110041383A (zh) | 2018-01-15 | 2018-01-15 | 硫酸软骨素寡糖、其制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110041383A (zh) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04225994A (ja) * | 1990-04-23 | 1992-08-14 | Akzo Nv | 三糖単位を含む炭水化物誘導体 |
| CN102676613A (zh) * | 2012-05-22 | 2012-09-19 | 江南大学 | 一种硫酸软骨素二糖、四糖、六糖的制备方法 |
| CN106467563A (zh) * | 2015-08-21 | 2017-03-01 | 北京大学 | 硫酸软骨素四糖的合成方法及其中间体化合物 |
-
2018
- 2018-01-15 CN CN201810033328.1A patent/CN110041383A/zh active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH04225994A (ja) * | 1990-04-23 | 1992-08-14 | Akzo Nv | 三糖単位を含む炭水化物誘導体 |
| CN102676613A (zh) * | 2012-05-22 | 2012-09-19 | 江南大学 | 一种硫酸软骨素二糖、四糖、六糖的制备方法 |
| CN106467563A (zh) * | 2015-08-21 | 2017-03-01 | 北京大学 | 硫酸软骨素四糖的合成方法及其中间体化合物 |
Non-Patent Citations (4)
| Title |
|---|
| JUN-ICHI TAMURA ET AL.: ""Synthesis and interaction with midkine of biotinylated chondroitin sulfate tetrasaccharides"", 《BIOORGANIC & MEDICINAL CHEMISTRY LETTERS》 * |
| KELLIE J. WILLIAMS ET AL.: ""Critical Elements of Oligosaccharide Acceptor Substrates for the Pasteurella multocida Hyaluronan Synthase"", 《JOURNAL OF BIOLOGICAL CHEMISTRY》 * |
| SHUANG YANG ET AL.: ""An investigation of construction of chondroitin sulfate E (CS-E) repeating unit"", 《TETRAHEDRON》 * |
| TED M. SLAGHEK ET AL.: ""Synthesis of hyaluronic acid related di- and tetra-saccharides having a glucuronic acid at the reducing end"", 《TETRAHEDRON: ASYMMETRY》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| ES2423888T3 (es) | Pentasacáridos de heparina sintéticos | |
| CN106536537B (zh) | 半乳凝素的新型杂合半乳糖苷抑制剂 | |
| Ning et al. | Synthesis of β-(1→ 6)-branched β-(1→ 3) glucohexaose and its analogues containing an α-(1→ 3) linked bond with antitumor activity | |
| CA2435363A1 (en) | New inhibitors against galectins | |
| JP5738272B2 (ja) | ６’−シアリルラクトース塩並びに６’−シアリルラクトース塩及び他のａ−シアリルオリゴ糖の合成方法 | |
| Iyer et al. | Design and synthesis of hyaluronan-mimetic gemini disaccharides | |
| Hollinger et al. | Synthesis of mucin O-glycan core structures as their p-nitro-and p-aminophenyl glycosides | |
| CN110041383A (zh) | 硫酸软骨素寡糖、其制备方法和应用 | |
| Kameyama et al. | A Total Synthesis of Sialyl Dimeric Lex Ganglioside1 | |
| CA2665392A1 (en) | Anticoagulant compounds | |
| AU2005276351A1 (en) | Azasugar derivatives, heparanase inhibitors, method for preparing same, compositions containing same, use thereof | |
| Juetten et al. | Stereoselective. alpha.-glycosylation of nitro sugar evernitrose: synthesis of the terminal AB unit of everninomicin antibiotics | |
| Jain et al. | Synthesis of the sodium salts of methyl 2-O-α-l-fucopyranosyl-α-l-fucopyranoside 3-and 4-sulfate | |
| Buskas et al. | Synthesis of oligosaccharides designed to form micelles, corresponding to structures found in ovarian cyst fluid | |
| CN112745372B (zh) | 一种基于tdg分子骨架的功能糖类分子及其制备方法 | |
| JP2001512737A (ja) | 置換されたテトラヒドロピラン誘導体、それらの製造方法、医薬または診断剤としてのそれらの使用およびそれらを含有する医薬 | |
| WO2021081881A1 (zh) | 一种基于tdg分子骨架的功能糖类分子及其制备方法 | |
| Chaudhury et al. | Synthesis of the pentasaccharide repeating unit of the O-antigen from Enterobacter cloacae C4115 containing the rare α-d-FucNAc | |
| CN104592319B (zh) | 一种糖胺聚糖类化合物n,o‑同时硫酸化的方法及其反应中间体 | |
| CN108884122B (zh) | 用于生产具有硫酸基和/或磷酸基的糖的方法 | |
| US6339064B1 (en) | Benzylglycosylamides as inhibitors of smooth muscle cell proliferation | |
| Shiu | Synthesis of [2.2. 2] L-Iduronate Lactone Thioglycoside and D-Galactosamine Derivatives Relevant to Glycosaminoglycan Fragments | |
| WO1998011118A1 (fr) | Derives x de lewis | |
| CN1594342B (zh) | 一类具有抗炎活性的糖基邻苯二甲酰亚胺类化合物 | |
| Hinou et al. | Synthesis of Amphiphilic Chitopentaose and Chitoheptaose Derivatives Using a Common Disaccharidic Synthon as the Chain Elongation Unit. |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20190723 |