CN106619674A - 栀子有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或保健食品中的应用 - Google Patents
栀子有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或保健食品中的应用 Download PDFInfo
- Publication number
- CN106619674A CN106619674A CN201610848914.2A CN201610848914A CN106619674A CN 106619674 A CN106619674 A CN 106619674A CN 201610848914 A CN201610848914 A CN 201610848914A CN 106619674 A CN106619674 A CN 106619674A
- Authority
- CN
- China
- Prior art keywords
- cape jasmine
- active component
- jasmine fruit
- aglycon
- alzheimer disease
- 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
- 208000024827 Alzheimer disease Diseases 0.000 title claims abstract description 92
- 239000003814 drug Substances 0.000 title claims abstract description 55
- 238000002360 preparation method Methods 0.000 title claims abstract description 25
- 235000013305 food Nutrition 0.000 title abstract description 10
- 241000207840 Jasminum Species 0.000 title abstract description 6
- 235000010254 Jasminum officinale Nutrition 0.000 title abstract description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 183
- 150000001875 compounds Chemical class 0.000 claims abstract description 140
- 238000010828 elution Methods 0.000 claims abstract description 127
- 235000013399 edible fruits Nutrition 0.000 claims abstract description 111
- 229930003658 monoterpene Natural products 0.000 claims abstract description 61
- 230000006386 memory function Effects 0.000 claims abstract description 36
- 229930003935 flavonoid Natural products 0.000 claims abstract description 26
- 150000002215 flavonoids Chemical class 0.000 claims abstract description 26
- 235000017173 flavonoids Nutrition 0.000 claims abstract description 26
- 239000000178 monomer Substances 0.000 claims abstract description 20
- 230000000694 effects Effects 0.000 claims abstract description 16
- 239000000463 material Substances 0.000 claims abstract description 11
- 239000011347 resin Substances 0.000 claims abstract description 9
- 229920005989 resin Polymers 0.000 claims abstract description 9
- 235000007586 terpenes Nutrition 0.000 claims abstract description 7
- 238000007670 refining Methods 0.000 claims abstract 7
- 150000003505 terpenes Chemical class 0.000 claims abstract 4
- 230000015572 biosynthetic process Effects 0.000 claims abstract 2
- 235000018958 Gardenia augusta Nutrition 0.000 claims description 113
- 244000111489 Gardenia augusta Species 0.000 claims description 110
- 229930182470 glycoside Natural products 0.000 claims description 93
- -1 Diterpenes glycosides Chemical class 0.000 claims description 81
- 235000002577 monoterpenes Nutrition 0.000 claims description 60
- DKVBOUDTNWVDEP-NJCHZNEYSA-N teicoplanin aglycone Chemical compound N([C@H](C(N[C@@H](C1=CC(O)=CC(O)=C1C=1C(O)=CC=C2C=1)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)OC=1C=C3C=C(C=1O)OC1=CC=C(C=C1Cl)C[C@H](C(=O)N1)NC([C@H](N)C=4C=C(O5)C(O)=CC=4)=O)C(=O)[C@@H]2NC(=O)[C@@H]3NC(=O)[C@@H]1C1=CC5=CC(O)=C1 DKVBOUDTNWVDEP-NJCHZNEYSA-N 0.000 claims description 44
- 229940079593 drug Drugs 0.000 claims description 35
- 150000002773 monoterpene derivatives Chemical class 0.000 claims description 34
- IBFYXTRXDNAPMM-BVTMAQQCSA-N Geniposide Chemical compound O([C@@H]1OC=C([C@@H]2[C@H]1C(=CC2)CO)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O IBFYXTRXDNAPMM-BVTMAQQCSA-N 0.000 claims description 31
- IBFYXTRXDNAPMM-FZEIBHLUSA-N Geniposide Natural products COC(=O)C1=CO[C@@H](O[C@H]2O[C@@H](CO)[C@H](O)[C@@H](O)[C@@H]2O)[C@H]2[C@@H]1CC=C2CO IBFYXTRXDNAPMM-FZEIBHLUSA-N 0.000 claims description 30
- VGLLGNISLBPZNL-RBUKDIBWSA-N arborescoside Natural products O=C(OC)C=1[C@@H]2C([C@H](O[C@H]3[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O3)OC=1)=C(CO)CC2 VGLLGNISLBPZNL-RBUKDIBWSA-N 0.000 claims description 30
- 235000013402 health food Nutrition 0.000 claims description 28
- 229930015704 phenylpropanoid Natural products 0.000 claims description 28
- 239000002253 acid Substances 0.000 claims description 24
- 239000004480 active ingredient Substances 0.000 claims description 24
- GIKVSFNAEBQLGB-UHFFFAOYSA-N 5,7-dimethoxy-2-(3,4,5-trimethoxyphenyl)chromen-4-one Chemical compound C=1C(OC)=CC(OC)=C(C(C=2)=O)C=1OC=2C1=CC(OC)=C(OC)C(OC)=C1 GIKVSFNAEBQLGB-UHFFFAOYSA-N 0.000 claims description 19
- VLCHQFXSBHIBRV-KNNWKXJLSA-N Mussaenosidic acid Natural products O=C(O)C=1[C@@H]2[C@H]([C@H](O[C@@H]3[C@@H](O)[C@H](O)[C@H](O)[C@@H](CO)O3)OC=1)[C@](O)(C)CC2 VLCHQFXSBHIBRV-KNNWKXJLSA-N 0.000 claims description 19
- FOONTNRMWNJWCL-PEZGRIKUSA-N Epijasminoside A Natural products O(C[C@@H]1C(C)=CC(=O)CC1(C)C)[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 FOONTNRMWNJWCL-PEZGRIKUSA-N 0.000 claims description 18
- BZPMXJKRKXDRID-UOIKKKDVSA-N Scandoside Natural products OC[C@H]1O[C@@H](O[C@H]2CC=C([C@@H]3[C@@H](O)C=C(CO)[C@H]23)C(=O)O)[C@H](O)[C@@H](O)[C@@H]1O BZPMXJKRKXDRID-UOIKKKDVSA-N 0.000 claims description 16
- 239000007924 injection Substances 0.000 claims description 16
- 238000002347 injection Methods 0.000 claims description 16
- SEBIKDIMAPSUBY-JAUCNNNOSA-N Crocin Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C(=O)OC1OC(COC2OC(CO)C(O)C(O)C2O)C(O)C(O)C1O)C=CC=C(/C)C(=O)OC3OC(COC4OC(CO)C(O)C(O)C4O)C(O)C(O)C3O SEBIKDIMAPSUBY-JAUCNNNOSA-N 0.000 claims description 14
- 150000004141 diterpene derivatives Chemical class 0.000 claims description 14
- 230000006993 memory improvement Effects 0.000 claims description 14
- FOONTNRMWNJWCL-UHFFFAOYSA-N Epijasminoside A Chemical compound CC1=CC(=O)CC(C)(C)C1COC1C(O)C(O)C(O)C(CO)O1 FOONTNRMWNJWCL-UHFFFAOYSA-N 0.000 claims description 13
- VLCHQFXSBHIBRV-NJPMDSMTSA-N Mussaenosidic acid Chemical compound O([C@H]1[C@H]2[C@@H](C(=CO1)C(O)=O)CC[C@@]2(O)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VLCHQFXSBHIBRV-NJPMDSMTSA-N 0.000 claims description 13
- 238000004440 column chromatography Methods 0.000 claims description 12
- 239000007788 liquid Substances 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 12
- VELYAQRXBJLJAK-XKKWFBPMSA-N (2s,3r,4s,5s,6r)-2-[[(1s,4ar,5r,7s,7as)-5,7-dihydroxy-7-methyl-4a,5,6,7a-tetrahydro-1h-cyclopenta[c]pyran-1-yl]oxy]-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O([C@@H]1OC=C[C@H]2[C@H](O)C[C@@]([C@@H]12)(O)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O VELYAQRXBJLJAK-XKKWFBPMSA-N 0.000 claims description 11
- RNEQSBBQXWZUJN-UHFFFAOYSA-N (4R)-4-{{6-O-[(2E)-3-(4-hydroxy-3,5-dimethoxyphenyl)-1-oxoprop-2-en-1-yl]-beta-D-glucopyranosyl}oxy}-2,6,6-trimethylcyclohex-1-ene-1-carboxylic acid Natural products COC1=C(O)C(OC)=CC(C=CC(=O)OCC2C(C(O)C(O)C(OC3CC(C)(C)C(C(O)=O)=C(C)C3)O2)O)=C1 RNEQSBBQXWZUJN-UHFFFAOYSA-N 0.000 claims description 10
- FLCVGMVLNHYJAW-UHFFFAOYSA-N 5-hydroxy-7-methoxy-2-(3,4,5-trimethoxyphenyl)chromen-4-one Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC(OC)=C(OC)C(OC)=C1 FLCVGMVLNHYJAW-UHFFFAOYSA-N 0.000 claims description 10
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 claims description 10
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 claims description 10
- KURIDZCRFQNRTO-UHFFFAOYSA-N Crocusatin C Natural products CC1=CC(=O)CC(C)(C)C1CO KURIDZCRFQNRTO-UHFFFAOYSA-N 0.000 claims description 10
- ZJDOESGVOWAULF-OGJQONSISA-N Geniposidic acid Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@@H]2C(CO)=CC[C@@H]2C(C(O)=O)=CO1 ZJDOESGVOWAULF-OGJQONSISA-N 0.000 claims description 10
- VYAALAFRWREWLA-BVTMAQQCSA-N Geniposidic acid Natural products CCC1=CC[C@H]2[C@@H]1[C@H](O[C@@H]3O[C@H](CO)[C@@H](O)[C@H](O)[C@H]3O)OC=C2C(=O)O VYAALAFRWREWLA-BVTMAQQCSA-N 0.000 claims description 10
- ZJDOESGVOWAULF-UHFFFAOYSA-N Geniposidinsaeure Natural products OC1C(O)C(O)C(CO)OC1OC1C2C(CO)=CCC2C(C(O)=O)=CO1 ZJDOESGVOWAULF-UHFFFAOYSA-N 0.000 claims description 10
- YSIFYNVXJOGADM-KDYWOABDSA-N Shanzhiside Chemical compound O([C@H]1[C@H]2[C@@H](C(=CO1)C(O)=O)[C@H](O)C[C@@]2(O)C)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O YSIFYNVXJOGADM-KDYWOABDSA-N 0.000 claims description 10
- PVPIPGMAEAJMTH-UHFFFAOYSA-N Shanzhiside Natural products OCC1OC(OC2OC=C(C3C(O)CC(O)C23)C(=O)O)C(O)C(O)C1O PVPIPGMAEAJMTH-UHFFFAOYSA-N 0.000 claims description 10
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 claims description 10
- KGDIDCUROGOWDM-WDALVHIASA-N (1s,4as,6s,7as)-6-hydroxy-7-methylidene-1-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4a,5,6,7a-tetrahydro-1h-cyclopenta[c]pyran-4-carbaldehyde Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@H]1[C@@H]2C(=C)[C@@H](O)C[C@@H]2C(C=O)=CO1 KGDIDCUROGOWDM-WDALVHIASA-N 0.000 claims description 9
- FOONTNRMWNJWCL-MZHQWRCYSA-N (4R)-3,5,5-trimethyl-4-[[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethyl]cyclohex-2-en-1-one Chemical compound CC1=CC(=O)CC(C)(C)[C@H]1CO[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O FOONTNRMWNJWCL-MZHQWRCYSA-N 0.000 claims description 9
- JPFJQHYDGYXING-SECBINFHSA-N (4s)-3,4-bis(hydroxymethyl)-5,5-dimethylcyclohex-2-en-1-one Chemical compound CC1(C)CC(=O)C=C(CO)[C@H]1CO JPFJQHYDGYXING-SECBINFHSA-N 0.000 claims description 9
- KURIDZCRFQNRTO-SECBINFHSA-N (4s)-4-(hydroxymethyl)-3,5,5-trimethylcyclohex-2-en-1-one Chemical compound CC1=CC(=O)CC(C)(C)[C@@H]1CO KURIDZCRFQNRTO-SECBINFHSA-N 0.000 claims description 9
- LQZNCRCYYWUMOL-UXXRCYHCSA-N 2,4,4-trimethyl-3-[[(2r,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethyl]cyclohex-2-en-1-one Chemical compound CC1(C)CCC(=O)C(C)=C1CO[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 LQZNCRCYYWUMOL-UXXRCYHCSA-N 0.000 claims description 9
- 229930190206 6'-O-Sinapoyljasminoside Natural products 0.000 claims description 9
- VELYAQRXBJLJAK-TYNWJONJSA-N Ajugol Natural products O([C@@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@@H]1OC=C[C@@H]2[C@H](O)C[C@@](O)(C)[C@@H]12 VELYAQRXBJLJAK-TYNWJONJSA-N 0.000 claims description 9
- XJMPAUZQVRGFRE-SCHFUKFYSA-N Gardenoside Natural products O=C(OC)C=1[C@H]2[C@H]([C@H](O[C@H]3[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O3)OC=1)[C@@](O)(CO)C=C2 XJMPAUZQVRGFRE-SCHFUKFYSA-N 0.000 claims description 9
- FOONTNRMWNJWCL-PMKSJIGOSA-N Jasminoside A Natural products O(C[C@H]1C(C)=CC(=O)CC1(C)C)[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 FOONTNRMWNJWCL-PMKSJIGOSA-N 0.000 claims description 9
- 230000002776 aggregation Effects 0.000 claims description 9
- 238000004220 aggregation Methods 0.000 claims description 9
- 230000032683 aging Effects 0.000 claims description 9
- QNTIUHXPTFEBJF-UHFFFAOYSA-N bornyl 6-O-beta-D-xylopyranosyl-beta-D-glucopyranoside Natural products CC1(C)C(C2)CCC1(C)C2OC(C(C(O)C1O)O)OC1COC1OCC(O)C(O)C1O QNTIUHXPTFEBJF-UHFFFAOYSA-N 0.000 claims description 9
- KGDIDCUROGOWDM-UHFFFAOYSA-N gardaloside Natural products OC1C(O)C(O)C(CO)OC1OC1C2C(=C)C(O)CC2C(C=O)=CO1 KGDIDCUROGOWDM-UHFFFAOYSA-N 0.000 claims description 9
- TXZBXNYXMDKPRV-UHFFFAOYSA-N gardoside methyl ester Natural products COC(O)C1=COC(OC2OC(CO)C(O)C(O)C2O)C3C1CC(O)C3=C TXZBXNYXMDKPRV-UHFFFAOYSA-N 0.000 claims description 9
- AFFNBNJOIDYUQN-UHFFFAOYSA-N gargoside methyl ester Natural products C1C(O)C(=C)C2C1C(C(=O)OC)=COC2OC1OC(CO)C(O)C(O)C1O AFFNBNJOIDYUQN-UHFFFAOYSA-N 0.000 claims description 9
- BNXFBCBJALNHOV-UHFFFAOYSA-N genameside D Natural products C1C=C(COC2C(C(O)C(O)C(CO)O2)O)C2C1C(C(=O)OC)=COC2OC1OC(CO)C(O)C(O)C1O BNXFBCBJALNHOV-UHFFFAOYSA-N 0.000 claims description 9
- IHFBPDAQLQOCBX-UHFFFAOYSA-N hispidulin Chemical compound C=1C(=O)C2=C(O)C(OC)=C(O)C=C2OC=1C1=CC=C(O)C=C1 IHFBPDAQLQOCBX-UHFFFAOYSA-N 0.000 claims description 9
- JPFJQHYDGYXING-UHFFFAOYSA-N jasminodiol Natural products CC1(C)CC(=O)C=C(CO)C1CO JPFJQHYDGYXING-UHFFFAOYSA-N 0.000 claims description 9
- XBRXLXOCHNGHBC-UHFFFAOYSA-N jasminoside B Natural products CC1(C)CC(=O)C=C(COC2OC(CO)C(O)C(O)C2O)C1CO XBRXLXOCHNGHBC-UHFFFAOYSA-N 0.000 claims description 9
- LQZNCRCYYWUMOL-UHFFFAOYSA-N jasminoside E Natural products CC1(C)CCC(=O)C(C)=C1COC1C(O)C(O)C(O)C(CO)O1 LQZNCRCYYWUMOL-UHFFFAOYSA-N 0.000 claims description 9
- KOMHTYYQRGPWLA-PEZGRIKUSA-N jasminoside G Natural products O(C[C@@H]1C(CO)=CC(=O)CC1(C)C)[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 KOMHTYYQRGPWLA-PEZGRIKUSA-N 0.000 claims description 9
- 230000015654 memory Effects 0.000 claims description 9
- XJMPAUZQVRGFRE-AYDWLWLASA-N methyl (1s,4as,7s,7as)-7-hydroxy-7-(hydroxymethyl)-1-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4a,7a-dihydro-1h-cyclopenta[c]pyran-4-carboxylate Chemical compound O([C@@H]1OC=C([C@@H]2[C@H]1[C@](C=C2)(O)CO)C(=O)OC)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O XJMPAUZQVRGFRE-AYDWLWLASA-N 0.000 claims description 9
- VELYAQRXBJLJAK-UHFFFAOYSA-N myoporoside Natural products C12C(C)(O)CC(O)C2C=COC1OC1OC(CO)C(O)C(O)C1O VELYAQRXBJLJAK-UHFFFAOYSA-N 0.000 claims description 9
- QULPQWKDDOBIOC-UHFFFAOYSA-N 5-hydroxy-6,7-dimethoxy-2-(3,4,5-trimethoxyphenyl)chromen-4-one Chemical compound COC1=C(OC)C(OC)=CC(C=2OC3=CC(OC)=C(OC)C(O)=C3C(=O)C=2)=C1 QULPQWKDDOBIOC-UHFFFAOYSA-N 0.000 claims description 8
- VWHCJIZYMBSOMV-UHFFFAOYSA-N 6''-O-trans-sinapoyl-(genipin gentiobioside) Natural products C1C=C(CO)C2C1C(C(=O)OC)=COC2OC(C(C(O)C1O)O)OC1COC(C(C(O)C1O)O)OC1COC(=O)C=CC1=CC(OC)=C(O)C(OC)=C1 VWHCJIZYMBSOMV-UHFFFAOYSA-N 0.000 claims description 8
- CSRRWPAMFHNUSX-XTOOWAAXSA-N 6''-O-trans-sinapoylgenipin gentiobioside Natural products COC(=O)C1=CO[C@@H](O[C@@H]2O[C@H](CO[C@@H]3O[C@H](COC(=O)C=Cc4ccc(O)c(O)c4)[C@@H](O)[C@H](O)[C@H]3O)[C@@H](O)[C@H](O)[C@H]2O)[C@H]5[C@@H]1CC=C5CO CSRRWPAMFHNUSX-XTOOWAAXSA-N 0.000 claims description 8
- 229930004069 diterpene Natural products 0.000 claims description 8
- CPCPHNWWTJLXKQ-UHFFFAOYSA-N 3',4',5'-O-trimethyltricetin Chemical compound COC1=C(OC)C(OC)=CC(C=2OC3=CC(O)=CC(O)=C3C(=O)C=2)=C1 CPCPHNWWTJLXKQ-UHFFFAOYSA-N 0.000 claims description 7
- WSGPLSDARZNMCW-UHFFFAOYSA-N Deacetylasperulosidsaeure-methylester Natural products OC1C=C(CO)C2C1C(C(=O)OC)=COC2OC1OC(CO)C(O)C(O)C1O WSGPLSDARZNMCW-UHFFFAOYSA-N 0.000 claims description 7
- KUSKILNIKZVCRA-UHFFFAOYSA-N jasminoside C Natural products CC1(C)CC(=O)C=C(COC2OC(CO)C(O)C(O)C2O)C1=C KUSKILNIKZVCRA-UHFFFAOYSA-N 0.000 claims description 7
- WFVABHHWJPWNJJ-UHFFFAOYSA-N 5-hydroxy-2-(3-hydroxy-4,5-dimethoxyphenyl)-7-methoxychromen-4-one Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC(O)=C(OC)C(OC)=C1 WFVABHHWJPWNJJ-UHFFFAOYSA-N 0.000 claims description 6
- VKWWRUZYBNTDHM-AXECJBQYSA-N Deacetylasperulosidic acid Natural products OCC1=C[C@H](O)[C@H]2[C@@H]1[C@H](OC[C@H]3O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]3O)OC=C2C(=O)O VKWWRUZYBNTDHM-AXECJBQYSA-N 0.000 claims description 6
- 238000009472 formulation Methods 0.000 claims description 6
- DLZFGILSEBIZHZ-UHFFFAOYSA-N jasminoside I Natural products CC1=C(COC2OC(COC3OC(CO)C(O)C(O)C3O)C(O)C(O)C2O)C(C)(C)CCC1=O DLZFGILSEBIZHZ-UHFFFAOYSA-N 0.000 claims description 6
- 239000000470 constituent Substances 0.000 claims description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 5
- 208000024172 Cardiovascular disease Diseases 0.000 claims description 4
- SEBIKDIMAPSUBY-ARYZWOCPSA-N Crocin Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O)OC(=O)C(C)=CC=CC(C)=C\C=C\C=C(/C)\C=C\C=C(C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)O1)O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O SEBIKDIMAPSUBY-ARYZWOCPSA-N 0.000 claims description 4
- 208000026139 Memory disease Diseases 0.000 claims description 4
- 239000004952 Polyamide Substances 0.000 claims description 4
- 102000054727 Serum Amyloid A Human genes 0.000 claims description 4
- 108700028909 Serum Amyloid A Proteins 0.000 claims description 4
- 239000002250 absorbent Substances 0.000 claims description 4
- 230000002745 absorbent Effects 0.000 claims description 4
- 230000006933 amyloid-beta aggregation Effects 0.000 claims description 4
- 208000026106 cerebrovascular disease Diseases 0.000 claims description 4
- 230000002526 effect on cardiovascular system Effects 0.000 claims description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 4
- 229930003647 monocyclic monoterpene Natural products 0.000 claims description 4
- 150000002767 monocyclic monoterpene derivatives Chemical class 0.000 claims description 4
- 229920002647 polyamide Polymers 0.000 claims description 4
- 230000002265 prevention Effects 0.000 claims description 4
- 208000019116 sleep disease Diseases 0.000 claims description 4
- 208000020685 sleep-wake disease Diseases 0.000 claims description 4
- 230000001629 suppression Effects 0.000 claims description 4
- SDWHLKFQMBNHOO-IVZWLZJFSA-N Gardenate A Natural products O=C(OC)[C@H](CO)[C@H]1[C@H](C(=O)OC)C(CO)=CC1 SDWHLKFQMBNHOO-IVZWLZJFSA-N 0.000 claims description 3
- 239000002775 capsule Substances 0.000 claims description 3
- 201000010099 disease Diseases 0.000 claims description 3
- 239000007938 effervescent tablet Substances 0.000 claims description 3
- 239000003826 tablet Substances 0.000 claims description 3
- 240000001972 Gardenia jasminoides Species 0.000 claims 4
- 230000006870 function Effects 0.000 claims 3
- GUJFIHUDZQDHLD-UHFFFAOYSA-N 3,5,6-trihydroxy-8-methoxy-2-phenylchromen-4-one Chemical compound COC1=CC(O)=C(O)C(C(C=2O)=O)=C1OC=2C1=CC=CC=C1 GUJFIHUDZQDHLD-UHFFFAOYSA-N 0.000 claims 1
- 241001529246 Platymiscium Species 0.000 claims 1
- 241001107098 Rubiaceae Species 0.000 claims 1
- 150000001336 alkenes Chemical class 0.000 claims 1
- 101800001718 Amyloid-beta protein Proteins 0.000 abstract 2
- 238000009825 accumulation Methods 0.000 abstract 1
- 229930182486 flavonoid glycoside Natural products 0.000 abstract 1
- 150000007955 flavonoid glycosides Chemical class 0.000 abstract 1
- 230000002401 inhibitory effect Effects 0.000 abstract 1
- 125000002950 monocyclic group Chemical group 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 182
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 148
- 238000005160 1H NMR spectroscopy Methods 0.000 description 60
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 45
- 210000004027 cell Anatomy 0.000 description 44
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 40
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 36
- 239000007787 solid Substances 0.000 description 35
- 230000001681 protective effect Effects 0.000 description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 28
- 239000000843 powder Substances 0.000 description 27
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 26
- 238000000034 method Methods 0.000 description 22
- 239000000741 silica gel Substances 0.000 description 20
- 229910002027 silica gel Inorganic materials 0.000 description 20
- 229960001866 silicon dioxide Drugs 0.000 description 20
- 241000700159 Rattus Species 0.000 description 19
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 19
- 230000003833 cell viability Effects 0.000 description 17
- 238000001514 detection method Methods 0.000 description 17
- 230000005779 cell damage Effects 0.000 description 15
- CZSBHMFVVLYIQQ-DRVLGOCHSA-N beta-D-gentiobiosyl beta-D-glucosyl crocetin Chemical compound O([C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)O1)O)C(=O)C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)C(=O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O CZSBHMFVVLYIQQ-DRVLGOCHSA-N 0.000 description 13
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 12
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 12
- 239000013078 crystal Substances 0.000 description 12
- 238000004128 high performance liquid chromatography Methods 0.000 description 12
- 238000011068 loading method Methods 0.000 description 12
- 239000012467 final product Substances 0.000 description 10
- OKKJLVBELUTLKV-VMNATFBRSA-N methanol-d1 Chemical compound [2H]OC OKKJLVBELUTLKV-VMNATFBRSA-N 0.000 description 10
- 239000012071 phase Substances 0.000 description 10
- 239000002904 solvent Substances 0.000 description 10
- 230000003595 spectral effect Effects 0.000 description 10
- CZSBHMFVVLYIQQ-RDVATZJHSA-N Crocin 2 Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C(=O)OC1OC(COC2OC(CO)C(O)C(O)C2O)C(O)C(O)C1O)C=CC=C(/C)C(=O)OC3OC(CO)C(O)C(O)C3O CZSBHMFVVLYIQQ-RDVATZJHSA-N 0.000 description 9
- 210000004556 brain Anatomy 0.000 description 9
- SEBIKDIMAPSUBY-RTJKDTQDSA-N crocin-1 Chemical compound C([C@H]1O[C@H]([C@@H]([C@@H](O)[C@@H]1O)O)OC(=O)C(/C)=C/C=C/C(/C)=C/C=C/C=C(\C)/C=C/C=C(\C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)O)O1)O)O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O SEBIKDIMAPSUBY-RTJKDTQDSA-N 0.000 description 9
- 238000005516 engineering process Methods 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 150000002338 glycosides Chemical class 0.000 description 9
- 230000006837 decompression Effects 0.000 description 8
- 239000012153 distilled water Substances 0.000 description 8
- 239000004615 ingredient Substances 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- 239000007921 spray Substances 0.000 description 8
- 238000001694 spray drying Methods 0.000 description 8
- JGHUOJAZXGSFRI-HOWDAYCMSA-N (4s,5z,6s)-4-(2-methoxy-2-oxoethyl)-5-[2-[(e)-3-phenylprop-2-enoyl]oxyethylidene]-6-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4h-pyran-3-carboxylic acid Chemical compound C1(/[C@@H](C(=CO[C@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)C(O)=O)CC(=O)OC)=C\COC(=O)\C=C\C1=CC=CC=C1 JGHUOJAZXGSFRI-HOWDAYCMSA-N 0.000 description 7
- OEZZJTAJYYSQKM-UHFFFAOYSA-N 5,7-dihydroxy-2-(4-hydroxyphenyl)-8-methoxychromen-4-one Chemical compound COC1=C(O)C=C(O)C(C(C=2)=O)=C1OC=2C1=CC=C(O)C=C1 OEZZJTAJYYSQKM-UHFFFAOYSA-N 0.000 description 7
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 7
- 230000009471 action Effects 0.000 description 7
- 230000036541 health Effects 0.000 description 7
- 238000011552 rat model Methods 0.000 description 7
- 239000013558 reference substance Substances 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- KMZQTSPTBCTYOF-UHFFFAOYSA-N 3,5,6-trihydroxy-2-(4-hydroxy-3,5-dimethoxyphenyl)chromen-4-one Chemical compound OC1=C(OC2=CC=C(C(=C2C1=O)O)O)C1=CC(=C(C(=C1)OC)O)OC KMZQTSPTBCTYOF-UHFFFAOYSA-N 0.000 description 6
- VWQSIGAFROAQLP-UHFFFAOYSA-N 3,6-dihydroxy-2,4,4-trimethylcyclohexene-1-carbaldehyde Chemical compound CC1=C(C(CC(C1O)(C)C)O)C=O VWQSIGAFROAQLP-UHFFFAOYSA-N 0.000 description 6
- 241000218636 Thuja Species 0.000 description 6
- 230000006378 damage Effects 0.000 description 6
- 239000003480 eluent Substances 0.000 description 6
- 239000000706 filtrate Substances 0.000 description 6
- 229930003944 flavone Natural products 0.000 description 6
- 150000002213 flavones Chemical class 0.000 description 6
- 235000011949 flavones Nutrition 0.000 description 6
- XGIUYVCQIWQCCU-UHFFFAOYSA-N 5,7-dihydroxy-2-(3-hydroxy-4,5-dimethoxyphenyl)-8-methoxychromen-4-one Chemical compound OC1=C(OC)C(OC)=CC(C=2OC3=C(OC)C(O)=CC(O)=C3C(=O)C=2)=C1 XGIUYVCQIWQCCU-UHFFFAOYSA-N 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
- 206010020772 Hypertension Diseases 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 5
- 230000002146 bilateral effect Effects 0.000 description 5
- 210000001168 carotid artery common Anatomy 0.000 description 5
- 239000006071 cream Substances 0.000 description 5
- 150000004702 methyl esters Chemical class 0.000 description 5
- 235000015097 nutrients Nutrition 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 230000003247 decreasing effect Effects 0.000 description 4
- 238000004992 fast atom bombardment mass spectroscopy Methods 0.000 description 4
- LHDWRKICQLTVDL-UHFFFAOYSA-N methyl iridoid glycoside Natural products OC1C(O)C(O)C(CO)OC1OC1C2C3(CO)OC3C(O)C2C=CO1 LHDWRKICQLTVDL-UHFFFAOYSA-N 0.000 description 4
- 230000010412 perfusion Effects 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- TVQCDZVYKULHJP-UHFFFAOYSA-N 4-hydroxy-2-(hydroxymethyl)-6,6-dimethylcyclohexene-1-carbaldehyde Chemical compound CC1(CC(CC(=C1C=O)CO)O)C TVQCDZVYKULHJP-UHFFFAOYSA-N 0.000 description 3
- 208000037259 Amyloid Plaque Diseases 0.000 description 3
- 206010002091 Anaesthesia Diseases 0.000 description 3
- 238000012347 Morris Water Maze Methods 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 125000002252 acyl group Chemical group 0.000 description 3
- 230000037005 anaesthesia Effects 0.000 description 3
- 230000003143 atherosclerotic effect Effects 0.000 description 3
- 230000000903 blocking effect Effects 0.000 description 3
- 230000017531 blood circulation Effects 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 3
- 239000003292 glue Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 208000019622 heart disease Diseases 0.000 description 3
- 210000001320 hippocampus Anatomy 0.000 description 3
- 230000001771 impaired effect Effects 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- JGHUOJAZXGSFRI-UHFFFAOYSA-N isojasminoside Natural products O1C(CO)C(O)C(O)C(O)C1OC1OC=C(C(O)=O)C(CC(=O)OC)C1=CCOC(=O)C=CC1=CC=CC=C1 JGHUOJAZXGSFRI-UHFFFAOYSA-N 0.000 description 3
- 229930188573 jasminoside Natural products 0.000 description 3
- 235000012054 meals Nutrition 0.000 description 3
- 210000002569 neuron Anatomy 0.000 description 3
- 239000002547 new drug Substances 0.000 description 3
- 230000006919 peptide aggregation Effects 0.000 description 3
- 230000000144 pharmacologic effect Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000010992 reflux Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- ZAIANDVQAMEDFL-UHFFFAOYSA-N 3-methoxy-2-phenylchromen-4-one Chemical compound O1C2=CC=CC=C2C(=O)C(OC)=C1C1=CC=CC=C1 ZAIANDVQAMEDFL-UHFFFAOYSA-N 0.000 description 2
- PTPQTMGOUIHFNR-UHFFFAOYSA-N 6-O-methylscandoside methyl ester Natural products COC(=O)C1=COC(OC2OC(CO)C(O)C(O)C2O)C3C1C(CO)C=C3CO PTPQTMGOUIHFNR-UHFFFAOYSA-N 0.000 description 2
- YWLXLRUDGLRYDR-LUPIKGFISA-N 7-epi-10-deacetylbaccatin iii Chemical group O([C@H]1[C@H]2[C@@](C([C@H](O)C3=C(C)[C@@H](O)C[C@]1(O)C3(C)C)=O)(C)[C@H](O)C[C@H]1OC[C@]12OC(=O)C)C(=O)C1=CC=CC=C1 YWLXLRUDGLRYDR-LUPIKGFISA-N 0.000 description 2
- 102000013455 Amyloid beta-Peptides Human genes 0.000 description 2
- 108010090849 Amyloid beta-Peptides Proteins 0.000 description 2
- 244000258271 Galium odoratum Species 0.000 description 2
- 235000008526 Galium odoratum Nutrition 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 208000028017 Psychotic disease Diseases 0.000 description 2
- 206010039966 Senile dementia Diseases 0.000 description 2
- 208000032109 Transient ischaemic attack Diseases 0.000 description 2
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 2
- 238000009098 adjuvant therapy Methods 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000008485 antagonism Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 210000003710 cerebral cortex Anatomy 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- ZSWFCLXCOIISFI-UHFFFAOYSA-N cyclopentadiene Chemical compound C1C=CC=C1 ZSWFCLXCOIISFI-UHFFFAOYSA-N 0.000 description 2
- 125000000567 diterpene group Chemical group 0.000 description 2
- 230000003181 encephalopathic effect Effects 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 238000003304 gavage Methods 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 235000008216 herbs Nutrition 0.000 description 2
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 2
- YEHDMSUNJUONMW-UHFFFAOYSA-N methoxyflavone Natural products COC1=CC=CC=C1C1=CC(=O)C2=CC=CC=C2O1 YEHDMSUNJUONMW-UHFFFAOYSA-N 0.000 description 2
- 210000005036 nerve Anatomy 0.000 description 2
- 239000002858 neurotransmitter agent Substances 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 230000007170 pathology Effects 0.000 description 2
- 229920000136 polysorbate Polymers 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 210000003625 skull Anatomy 0.000 description 2
- 238000002798 spectrophotometry method Methods 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 230000009182 swimming Effects 0.000 description 2
- 238000002636 symptomatic treatment Methods 0.000 description 2
- ULSUXBXHSYSGDT-UHFFFAOYSA-N tangeretin Chemical compound C1=CC(OC)=CC=C1C1=CC(=O)C2=C(OC)C(OC)=C(OC)C(OC)=C2O1 ULSUXBXHSYSGDT-UHFFFAOYSA-N 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 201000010875 transient cerebral ischemia Diseases 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- 230000002792 vascular Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 239000012224 working solution Substances 0.000 description 2
- ZQPVHVKWCGZNDW-NVYKSAHZSA-N (2r,3s,4s,5r,6r)-2-(hydroxymethyl)-6-[[(2r,3s,4s,5r,6r)-3,4,5-trihydroxy-6-methoxyoxan-2-yl]methoxy]oxane-3,4,5-triol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](OC)O[C@@H]1CO[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 ZQPVHVKWCGZNDW-NVYKSAHZSA-N 0.000 description 1
- KSEBMYQBYZTDHS-HWKANZROSA-M (E)-Ferulic acid Natural products COC1=CC(\C=C\C([O-])=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-M 0.000 description 1
- MLYYOHQNXPDGGV-UHFFFAOYSA-N 3,5-dimethoxy-2-phenylchromen-4-one Chemical compound COC=1C(=O)C=2C(OC)=CC=CC=2OC=1C1=CC=CC=C1 MLYYOHQNXPDGGV-UHFFFAOYSA-N 0.000 description 1
- JNBQPUBXNVAICI-UHFFFAOYSA-N 4-hydroxy-3,3-dimethyl-1-propan-2-yl-7-oxabicyclo[4.1.0]hept-4-en-2-one Chemical compound CC(C)C12C(O1)C=C(C(C2=O)(C)C)O JNBQPUBXNVAICI-UHFFFAOYSA-N 0.000 description 1
- YMYJODGCOOQALU-UHFFFAOYSA-N 6-hydroxy-3-(hydroxymethyl)-4,4-dimethylcyclohexene-1-carbaldehyde Chemical compound CC1(CC(C(=CC1CO)C=O)O)C YMYJODGCOOQALU-UHFFFAOYSA-N 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 201000006474 Brain Ischemia Diseases 0.000 description 1
- 206010008120 Cerebral ischaemia Diseases 0.000 description 1
- 101800004637 Communis Proteins 0.000 description 1
- 206010012289 Dementia Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- RQRMWYOWQUEKAS-UHFFFAOYSA-N Gardendiol Natural products OCC1C2CC=C(CO)C2COC1=O RQRMWYOWQUEKAS-UHFFFAOYSA-N 0.000 description 1
- JSKCJJNYSGWZDU-PKPQBBKFSA-N Gardoside Natural products O=C(O)C=1[C@@H]2[C@H]([C@H](O[C@H]3[C@@H](O)[C@H](O)[C@H](O)[C@@H](CO)O3)OC=1)C(=C)[C@@H](O)C2 JSKCJJNYSGWZDU-PKPQBBKFSA-N 0.000 description 1
- 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 description 1
- VAYOSLLFUXYJDT-RDTXWAMCSA-N Lysergic acid diethylamide Chemical compound C1=CC(C=2[C@H](N(C)C[C@@H](C=2)C(=O)N(CC)CC)C2)=C3C2=CNC3=C1 VAYOSLLFUXYJDT-RDTXWAMCSA-N 0.000 description 1
- 208000012902 Nervous system disease Diseases 0.000 description 1
- RTYFDBFRZMAENF-UHFFFAOYSA-N OC1=C2C(C=C(OC2=CC(=C1)O)C1=CC(=C(C(=C1)OC)OC)OC)=O.OC1=C2C(C=C(OC2=CC(=C1)O)C1=CC(=C(C(=C1)OC)OC)OC)=O Chemical compound OC1=C2C(C=C(OC2=CC(=C1)O)C1=CC(=C(C(=C1)OC)OC)OC)=O.OC1=C2C(C=C(OC2=CC(=C1)O)C1=CC(=C(C(=C1)OC)OC)OC)=O RTYFDBFRZMAENF-UHFFFAOYSA-N 0.000 description 1
- 208000006735 Periostitis Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 201000004810 Vascular dementia Diseases 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 238000001467 acupuncture Methods 0.000 description 1
- 125000004423 acyloxy group Chemical group 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 206010008118 cerebral infarction Diseases 0.000 description 1
- 229940126678 chinese medicines Drugs 0.000 description 1
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
- 229960002327 chloral hydrate Drugs 0.000 description 1
- 210000004081 cilia Anatomy 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- NBGJGWFIDMDCAW-UHFFFAOYSA-N egonol-beta-gentiobioside Natural products C=1C=2C=C(C=3C=C4OCOC4=CC=3)OC=2C(OC)=CC=1CCCOC(C(C(O)C1O)O)OC1COC1OC(CO)C(O)C(O)C1O NBGJGWFIDMDCAW-UHFFFAOYSA-N 0.000 description 1
- 238000002451 electron ionisation mass spectrometry Methods 0.000 description 1
- 150000002118 epoxides Chemical class 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- KSEBMYQBYZTDHS-HWKANZROSA-N ferulic acid Chemical group COC1=CC(\C=C\C(O)=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-N 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- WSFSSNUMVMOOMR-UHFFFAOYSA-N formaldehyde Substances O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 1
- 229960004279 formaldehyde Drugs 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- AZKVWQKMDGGDSV-UHFFFAOYSA-N genipin Natural products COC(=O)C1=COC(O)C2C(CO)=CCC12 AZKVWQKMDGGDSV-UHFFFAOYSA-N 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000001951 hemoperfusion Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 229930182489 iridoid glycoside Natural products 0.000 description 1
- 150000008145 iridoid glycosides Chemical class 0.000 description 1
- 230000000302 ischemic effect Effects 0.000 description 1
- OAUMGPGYHSGESA-UHFFFAOYSA-N jasminoside D Natural products CC1=C(C=O)C(C)(C)CC(O)C1OC2OC(CO)C(O)C(O)C2O OAUMGPGYHSGESA-UHFFFAOYSA-N 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000003446 memory effect Effects 0.000 description 1
- 125000001160 methoxycarbonyl group Chemical group [H]C([H])([H])OC(*)=O 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 150000004880 oxines Chemical class 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 210000003460 periosteum Anatomy 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229950008882 polysorbate Drugs 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 230000000384 rearing effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 230000010410 reperfusion Effects 0.000 description 1
- 235000014102 seafood Nutrition 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 229940036051 sojourn Drugs 0.000 description 1
- 230000006886 spatial memory Effects 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- HTSABYAWKQAHBT-UHFFFAOYSA-N trans 3-methylcyclohexanol Natural products CC1CCCC(O)C1 HTSABYAWKQAHBT-UHFFFAOYSA-N 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/74—Rubiaceae (Madder family)
- A61K36/744—Gardenia
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Epidemiology (AREA)
- Alternative & Traditional Medicine (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及栀子果实有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用。所制备栀子果实药材上述有效部位或有效成分本身是栀子果实总提物上大孔吸附树脂的30%或60%乙醇洗脱部位并具有治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品,及其30%或60%乙醇洗脱有效部位分离纯化精制得到萜类或黄酮类单体化合物并具有抑制Aβ纤丝形成和阻止其聚集的生物活性。因拥有共同的环烯醚萜类苷元、单环单萜类苷元、黄酮类苷元等相同或相似母核结构单元,而具有类似的治疗阿尔茨海默病或具有辅助改善记忆的功效。
Description
技术领域
本发明涉及栀子果实有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用,属于中医药制备技术领域。
背景技术
阿尔茨海默病(Alzheimer's Disease,AD)是一种神经系统退行性疾病,老年人发病率较高,临床主要表现为记忆力减退、定向障碍和判断推理能力受损等。现在比较公认AD(即老年期痴呆)的分类包括老年性痴呆、血管性痴呆和混合性痴呆。老年斑是AD的主要病理标志,其中Aβ聚集是中心环节,也是AD发病的关键性因素和关键性治疗靶点,干扰Aβ的产生和阻止其聚集是防治AD的有效途径,开发抑制Aβ纤丝形成和聚集的新药对预防和对症治疗AD具有良好的学术与临床应用价值。
本发明从动物因素诱发性与AD临床病因病机相结合的异质性及多因性出发,将脑衰老及血管危险因子(高血压、动脉粥样硬化和短暂性脑缺血等)导致的脑低灌注和AD脑病理病变相结合,利用腹腔注射半乳糖促老化与反复结扎双侧颈总动脉致脑低灌注及鼠脑室一次性注射β淀粉样蛋白制作异质性及多因性阿尔茨海默病(Heterogeneitys/Multi‐factors'Alzheimer Disease,H/MAD)模型或记忆力减退模型。研究证实栀子果实总提物上大孔吸附树脂的30%或60%乙醇洗脱部位为治疗异质性及多因性阿尔茨海默病的有效部位或辅助改善记忆功效,从其栀子果实30%或60%乙醇洗脱有效部位中分离鉴定100多个化合物,应用Aβ25-35损伤的PC12细胞模型对分离得到的栀子有效部位中的单体化合物进行活性筛选,其中栀子果实30%乙醇洗脱有效部位中的单萜苷类单体化合物具有抑制Aβ纤丝形成和阻止其聚集的生物活性,另外栀子果实60%乙醇洗脱有效部位中的单萜类苷元、二萜苷类、环烯醚萜苯丙素苷类和黄酮类成分单体化合物具有抑制Aβ纤丝形成和阻止其聚集的生物活性。上述有效部位或有效成分因拥有共同的环烯醚萜类苷元、单环单萜类苷元、黄酮类苷元等相同或相似母核结构单元,而具有类似的治疗阿尔茨海默病或辅助改善记忆的功效。上述研究还没有报道,我们通过系统的体内外实验研究,首先发现栀子果实有效部位或有效成分对由衰老、睡眠障碍、心脑血管疾病(心脏病、高血压、动脉粥样硬化和短暂性脑缺血发作等)、神经系统疾病等导致的淀粉样蛋白Aβ沉积或Aβ纤丝聚集形成的阿尔茨海默病疾病或记忆障碍或记忆力减退具有良好的治疗作用或辅助治疗功效。
发明内容
栀子果实有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用。
本发明采用了下述技术方案。
栀子果实有效部位或有效成分的制备方法:将栀子干燥果实粉碎成粗粉,用70%乙醇浸泡1‐2小时,4‐10倍量溶剂加热回流提取2‐4次,每次1‐3小时,过滤,合并滤液。上述滤液与45‐80℃减压浓缩至0.8‐1.2g生药材/ml,0‐4℃冷藏过夜,析胶,过滤,滤液直接上大孔吸附树脂AB‐8型或D‐101型或HPD‐400或各种型号大孔吸附树脂富集,依次用水、30%乙醇、60%乙醇、95%乙醇洗脱,得到各洗脱部位,主要收集30%和60%乙醇水洗脱部位,再将30%乙醇水洗脱物经聚酰胺柱色谱纯化得总单萜类化合物(总含量大于90%),并将该部位继续分离得到单体京尼平苷(含量98%)。各洗脱部位在45‐65℃减压回收溶剂至乙醇适量浓度,用冷冻干燥法或喷雾干燥进行干燥,收集冻干粉或喷雾干燥粉,即得。
栀子果实30%乙醇水洗脱有效部位中单萜苷类有效成分京尼平苷、去乙酰车叶草苷酸甲酯、gardaloside、6-甲氧基鸡屎藤次苷甲酯、6-甲氧基去乙酰车叶草苷酸甲酯、栀子苷、ajugol、genipin-1-β-gentiobiside、genipin-1,10-di-O-β-D-glucopyranoside、gardoside methyl ester、shanzhiside、Lamalbidicacid、4-carboxyl-7-hydroxy-catapol acid、4-carboxyl-catapol acid、Mussaenosidic acid、geniposidic acid、6-hydroxymethy-jasminoside D、bornyl-6-O-β-D-xylopyranosyl-β-D-glucopyranoside、jasminoside A、epijasminoside A、jasminoside B、jasminoside E、jasminoside G、jasminoside I的制备方法:将权利要求1-10所得栀子果实30%乙醇水洗脱有效部位上硅胶柱,上样后依次用三氯甲烷:甲醇或二氯甲烷:甲醇或三氯甲烷:甲醇:水或二氯甲烷:甲醇:水梯度洗脱,以上述栀子果实30%乙醇水洗脱有效部位中单萜苷类的相应化合物单体作为对照品TLC或HPLC检测,合并相应流份。上述流份于45‐65℃减压浓缩蒸干,用适量蒸馏水或适当溶剂混悬或溶解,上反相硅胶ODS‐C18柱色谱或SephadexLH‐20柱色谱或制备高效液相色谱技术分离纯化,流动相为甲醇水5-100%或乙腈水5-100%的等度或梯度洗脱,洗脱液用冷冻干燥法或喷雾干燥进行干燥,收集冻干粉或喷雾干燥粉,用95%乙醇结晶并重结晶,即得。
栀子60%乙醇洗脱有效部位中单萜类苷元成分jasminoside C aglycone、5,5-dihydroxymethyl-1-cyclohexene-furan lactone、6-hydroxyl-3-hydroxy-methyl-4,4-dimethyl-1-cyclohexene-1-formaldehyde、4-hydroxyl-2-hydroxymethyl-6,6-dime-thyl-1-cyclohexene-1-formaldehyde、4-hydroxyl-1,5,5-trimethyl-6-epoxyethyl-cyclo-hex-3-enone、iridoids 2a、iridoids 2b、jasminodiol、crocusatin-C、7-Epi-crocusatin-C、(10S,11S)-Gardendiol、(10R,11R)-Gardendiol)、(5S,9S)-Gardenate A、(5R,9R)-Gardenate A、5,6-dihydroxymethyl-1,1-dimethylcyclohex-4-enone)、1-hydroxy-7-hydroxylmethyl-1,4a,5,7a-tetrahydro-pyrancyclopenta-4-carbaldehyde或环烯醚萜苯丙素苷类成分5-aldehyde-6′-O-sinapoyljasminoside A、6′-O-sinapoyljasminoside B、6′-O-trans-sinapoyljasminoside L、6′-O-sinapoyl-3-methoxy-4-deketone-jasminoside E、6″-O-trans-sina-poylgenipin gentiobioside、6″-O-[(E)-p-coumaroyl]-gentiobiosyl-genipin、6″-O-[3″′-methoxy-(E)-caffeoyl]-gentiobiosylgenipin、4″-O-[(E)-p-coumar-oyl]-gentiobiosylgenipin、6″-O-[(E)-feruloyl]-gentiobiosylgenipin、6″-O-[(E)-cinnam-oyl]-gentiobiosylgenipin或黄酮类成分5,7-dihydroxyl-3′,4′,5′-trimethoxyflavone、5,7,4′-trihydroxyl-6-methoxyflavone、5,7,4′-trihydroxyl-8-methoxyflavone、5,7,3′-trihydroxyl-8,4′,5′-trimethoxyflavone、5-hydroxyl-6,7,3′,4′,5′-pentamethoxyflavone、5,3′-dihydroxyl-7,4′,5′-trimethoxyflavone、5-hydroxyl-7,3′,4′,5′-tetramethoxyflavone、3,5,6,4′-tetrahydroxy-3′,5′-dimethoxyflavone、5,7,3′,4′,5′-pentame-thoxyflavone或二萜类成分crocin-I、crocin-II的制备方法:将权利要求1-10所得栀子果实60%乙醇水洗脱有效部位上硅胶柱,上样后依次用三氯甲烷:甲醇或二氯甲烷:甲醇或三氯甲烷:甲醇:水或二氯甲烷:甲醇:水梯度洗脱,以上述栀子果实60%乙醇水洗脱有效部位中单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分的相应化合物单体作为对照品TLC或HPLC检测,合并相应流份。上述流份于45‐65℃减压浓缩蒸干,用适量蒸馏水或适当溶剂混悬或溶解,上反相硅胶ODS‐C18柱色谱或SephadexLH‐20柱色谱或制备高效液相色谱技术分离纯化,流动相为甲醇水5-100%或乙腈水5-100%的等度或梯度洗脱,洗脱液用冷冻干燥法或喷雾干燥进行干燥,收集冻干粉或喷雾干燥粉,用95%乙醇结晶并重结晶,即得。
前述方法制备的栀子果实有效部位中有效成分为京尼平苷、去乙酰车叶草苷酸甲酯、gardaloside、6-甲氧基鸡屎藤次苷甲酯、6-甲氧基去乙酰车叶草苷酸甲酯、栀子苷、ajugol、genipin-1-β-gentiobiside、genipin-1,10-di-O-β-D-glucopyrano-side、gardoside methyl ester、shanzhiside、Lamalbidicacid、4-carboxyl-7-hydroxy-catapol acid、4-carboxyl-catapol acid、Mussaenosidic acid、geniposidic acid、6-hydroxymethy-jasminoside D、bornyl-6-O-β-D-xylopyranosyl-β-D-glucopyrano-side、jasminoside A、epijasminoside A、jasminoside B、jasminoside E、jasminosideG、jasminoside I、jasminoside C aglycone、5,5-dihydroxymethyl-1-cyclohexene-furan lactone、6-hydroxyl-3-hydroxy-methyl-4,4-dimethyl-1-cyclo-hexene-1-formaldehyde、4-hydroxyl-2-hydroxymethyl-6,6-dimethyl-1-cyclohexene-formaldehyde、4-hydroxyl-1,5,5-trimethyl-6-epoxyethyl-cyclohex-3-enone、iridoids 2a、iridoids 2b、jasminodiol、crocusatin-C、7-Epi-crocusatin-C、(10S,11S)-Gardendiol、(10R,11R)-Gardendiol)、(5S,9S)-Gardenate A、(5R,9R)-GardenateA、5,6-dihydroxy-methyl-1,1-dimethylcyclohex-4-enone)、1-hydroxy-7-hydroxylmethyl-1,4a,5,7a-tetrahydro-pyrancyclopenta-4-carbaldehyde、5-aldehyde-6′-O-sinapoyljasminoside A、6′-O-sinapoyljasminoside B、6′-O-trans-sinapoyljasminoside L、6′-O-sinapoyl-3-methoxy-4-deketone-jasminoside E、6″-O-trans-sina-poylgenipin gentiobioside、6″-O-[(E)-p-coumaroyl]-gentiobiosyl-genipin、6″-O-[3″′-methoxy-(E)-caffeoyl]-gentiobiosylgenipin、4″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin、6″-O-[(E)-ferul-oyl]-gentiobiosylgenipin、6″-O-[(E)-cinnamoyl]-gentiobiosylgenipin、5,7-dihydroxyl-3′,4′,5′-trimethoxyflavone、5,7,4′-trihydroxyl-6-methoxyflavone、5,7,4′-trihydroxyl-8-methoxyflavone、5,7,3′-trihydroxyl-8,4′,5′-trimethoxyflavone、5-hydroxyl-6,7,3′,4′,5′-pentamethoxyflavone、5,3′-dihydroxyl-7,4′,5′-trimethoxyflavone、5-hydroxyl-7,3′,4′,5′-tetramethoxyflavone、3,5,6,4′-tetrahydroxy-3′,5′-dimethoxyflavone、5,7,3′,4′,5′-pentame-thoxyflavone、crocin-I、crocin-II中的一种或多种可用于制备治疗阿尔茨海默病的药物或辅助改善记忆功能保健食品,所述药物包括口服液、胶囊、片剂、泡腾片、注射剂或各种已知剂型制剂或可接受剂型制剂,所述药物还包括各种单方和复方制剂,各种已知或各种可接受辅助改善记忆功能类型保健食品。所涉及的阿尔茨海默病或记忆力减退是由衰老、睡眠障碍、心脑血管疾病(心脏病、高血压、动脉粥样硬化和短暂性脑缺血发作等)、精神系统疾病导致的淀粉样蛋白Aβ沉积或Aβ纤丝聚集形成的阿尔茨海默病疾病或记忆障碍或记忆力减退中的任意一种。
本发明的优点是:提供了栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中有效部位或有效成分的提取与制备方法,所得有效部位或有效成分可用于制备治疗异质性及多因性阿尔茨海默病或辅助改善记忆功能保健食品及其在制备各类应用药物的口服液、胶囊、片剂、泡腾片、粉针剂、水针剂、注射剂或各种已知剂型制剂或各种可接受剂型制剂的原料药。
栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的有效部位或有效成分在体内外对由衰老、睡眠障碍、心脑血管疾病(心脏病、高血压、动脉粥样硬化和短暂性脑缺血发作等)、精神系统疾病导致的淀粉样蛋白Aβ沉积或Aβ纤丝聚集形成的阿尔茨海默病或记忆障碍或记忆力减退具有良好的治疗作用和较强的生物活性或辅助治疗功效。
栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的有效部位或有效成分的制备方法主要通过从天然药物中提取,为纯天然组分与单体,具有良好的生物相容性。其中,有效成分6-hydroxymethy-jasminoside D、jasminoside Caglycone(1,1-dimethyl-5-hydroxy-methyl-6-vinyl-cyclohex-3-enone)、5,5-dihydroxymethyl-1-cyclohexene-furan lactone、6-hydroxyl-3-hydroxy-methyl-4,4-dimethyl-1-cyclohexene-1-formaldehyde、4-hydroxyl-2-hydroxymethyl-6,6-dimethyl-1-cyclohexene-1-formaldehyde、4-hydroxyl-1,5,5-trimethyl-6-epoxy-ethyl-cyclohex-3-enone、5-aldehyde-6′-O-sinapoyljasminoside A、6′-O-sinapoyljasm-inoside B、6′-O-trans-sinapoyljasminoside L、6′-O-sinapoyl-3-methoxy-4-deketone-jasminoside E为新化合物。
附图说明
图1为不同浓度的单萜苷类单体化合物Z1~Z8对受损的PC12细胞的保护作用。
图2为不同浓度的单萜苷类单体化合物Z9~Z16对受损的PC12细胞的保护作用。
图3为不同浓度的单萜苷类单体化合物Z17~Z24对受损的PC12细胞的保护作用。
图4为不同浓度的单萜类苷元单体化合物Z25~Z30对受损的PC12细胞的保护作用。
图5为不同浓度的单萜类苷元单体化合物Z31~Z38对受损的PC12细胞的保护作用。
图6为不同浓度的环烯醚萜类苯丙素苷单体化合物Z39~Z48对受损的PC12细胞的保护作用。
图7为不同浓度的黄酮类单体化合物Z49~Z57对受损的PC12细胞的保护作用。
图8为不同浓度的藏红花素二萜类单体化合物Z58~Z59对受损的PC12细胞的保护作用。
具体实施方式
下面结合实施例对本发明作进一步描述:
实施例1
栀子果实药材提取物及30%或60%乙醇洗脱部位的制备方法:栀子果实药材粉碎成粗粉,用70%乙醇浸泡1小时,10倍量溶剂加热回流提取3次,每次2小时,过滤,合并滤液。上述滤液于50℃减压浓缩至1.0g生药材/ml,0-4℃冷藏过夜,析胶,过滤,滤液上AB-8型号大孔吸附树脂富集,依次用水、30%乙醇、60%乙醇和95%乙醇洗脱,得到各洗脱部位,主要收集30%和60%乙醇水洗脱部位,再将30%乙醇水洗脱物经聚酰胺柱色谱纯化得总单萜类化合物(总含量大于90%),并将该部位继续分离得到单体京尼平苷(含量98%),用冷冻干燥法进行干燥,收集冻干粉,即得。其中栀子果实30%的洗脱部位以单萜苷类成分为主,单萜苷类总含量大于90%。另外,栀子果实60%乙醇洗脱部位以单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分为主,其上述成分总含量大于90%。即得栀子果实药材提取物或30%乙醇洗脱部位或60%乙醇洗脱部位。
实施例2
栀子果实30%乙醇洗脱有效部位以单萜苷类成分为主,其总含量大于90%,该栀子果实30%乙醇洗脱有效部位中有效成分京尼平苷、去乙酰车叶草苷酸甲酯、gardaloside、6-甲氧基鸡屎藤次苷甲酯、6-甲氧基去乙酰车叶草苷酸甲酯、栀子苷、ajugol、genipin-1-β-gentiobiside、genipin-1,10-di-O-β-D-glucopyranoside、gardoside methyl ester、shanzhiside、Lamalbidicacid、4-carboxyl-7-hydroxy-catapol acid、4-carboxyl-catapol acid、Mussaenosidic acid、geniposidic acid、6-hydroxymethy-jasminoside D、bornyl-6-O-β-D-xylopyranosyl-β-D-glucopyran-oside、jasminoside A、epijasminoside A、jasminoside B、jasminoside E、jasminoside G、jasminoside I中的一种或多种单萜类单体化合物的制备方法:将上述实例1中得到的30%乙醇洗脱部位的浸膏加入等量的100-200目硅胶拌匀,50℃减压挥干溶剂,研匀,干法上样,上常压硅胶柱(200-300目,硅胶量为浸膏的30倍),上样后依次用三氯甲烷:甲醇梯度洗脱,主要收集三氯甲烷:甲醇为6:1至2:1的洗脱流份,以上述栀子果实30%乙醇洗脱有效部位中的单萜苷类单体化合物作为对照品TLC或HPLC检测,合并相应流份。上述流份于50℃减压浓缩蒸干,用适量蒸馏水混悬,上SephadexLH‐20柱色谱或用制备高效液相色谱技术分离纯化,依次用乙醇水由0‐50%洗脱,再用乙醇水50‐95%洗脱,洗脱液用喷雾干燥进行干燥,收集喷雾干燥粉,用95%乙醇结晶并重结晶,即得。采用上述方法制备栀子果实30%乙醇洗脱有效部位中的单萜苷类单体化合物为0.5-25g/kg,经HPLC检测纯度为98-99.1%;色谱条件UItimate○RXB-C18色谱柱(4.6mm×150mm,3μm),流动相为乙腈-水(10%-100%梯度洗脱),检测波长为238或240nm,流速为1.0ml/min,柱温30℃,进样量20μl。
实施例3
栀子果实60%乙醇洗脱有效部位以单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分为主,其上述成分总含量大于90%,该栀子果实60%乙醇洗脱有效部位中有效成分jasminoside C aglycone、5,5-dihydroxymethyl-1-cyclohexe-ne-furan lactone、6-hydroxyl-3-hydroxy-methyl-4,4-dimethyl-1-cyclohexene-1-for-maldehyde、4-hydroxyl-2-hydroxymethyl-6,6-dimethyl-1-cyclohexene-1-formalde-hyde、4-hydroxyl-1,5,5-trimethyl-6-epoxyeth-yl-cyclohex-3-enone、iridoids 2a、iridoids2b、jasminodiol、crocusatin-C、7-Epi-crocusatin-C、(10S,11S)-Gardendiol、(10R,11R)-Gardendiol、(5S,9S)-Garde-nate A、(5R,9R)-Gardenate A、5,6-dihydroxy-methyl-1,1-dimethylcyclohex-4-enone、1-hydroxy-7-hydroxylmethyl-1,4a,5,7a-tetra-hydro-pyrancyclopenta-4-carbaldehyde中的一种或多种单萜类苷元单体化合物的制备方法:将上述实例1中得到的60%乙醇洗脱部位的浸膏加入等量的100-200目硅胶拌匀,50℃减压挥干溶剂,研匀,干法上样,上常压硅胶柱(200-300目,硅胶量为浸膏的40倍),上样后依次用三氯甲烷:甲醇梯度洗脱,主要收集三氯甲烷:甲醇为12:1至8:1的洗脱流份,以上述栀子果实60%乙醇洗脱有效部位中的单萜类苷元单体化合物作为对照品TLC或HPLC检测,合并相应流份。上述流份于50℃减压浓缩蒸干,用适量蒸馏水混悬,上SephadexLH‐20柱色谱或用制备高效液相色谱技术分离纯化,依次用乙醇水由0‐30%洗脱,再用乙醇水30‐95%洗脱,洗脱液用喷雾干燥进行干燥,收集喷雾干燥粉,用95%乙醇结晶并重结晶,即得。采用上述方法制备栀子果实60%乙醇洗脱有效部位中的单萜类苷元单体化合物为0.5-6.8g/kg,经HPLC检测纯度为97-98.6%;色谱条件UItimate○RXB-C18色谱柱(4.6mm×150mm,3μm),流动相为乙腈-水(10%-100%梯度洗脱),检测波长为238或245nm,流速为1.0ml/min,柱温30℃,进样量20μl。
实施例4
栀子果实60%乙醇洗脱有效部位以单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分为主,其上述成分总含量大于90%,该栀子果实60%乙醇洗脱有效部位中有效成分5-aldehyde-6′-O-sinapoyljasminoside A、6′-O-sinapoyljasmin-oside B、6′-O-trans-sinapoyljasminoside L、6′-O-sinapoyl-3-methoxy-4-deketone-jasminoside E、6″-O-trans-sinapoylgenipin gentiobioside、6″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin、6″-O-[3″′-methoxy-(E)-caffeoyl]-gentiobiosylgenipin、4″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin、6″-O-[(E)-feruloyl]-gentiobiosylgenipin、6″-O-[(E)-cinnamoyl]-gentiobiosylgenipin中的一种或多种环烯醚萜苯丙素苷类单体化合物的制备方法:将上述实例1中得到的60%乙醇洗脱部位的浸膏加入等量的100-200目硅胶拌匀,50℃减压挥干溶剂,研匀,干法上样,上常压硅胶柱(200-300目,硅胶量为浸膏的40倍),上样后依次用三氯甲烷:甲醇梯度洗脱,主要收集三氯甲烷:甲醇为7:1至3:1的洗脱流份,以上述栀子果实60%乙醇洗脱有效部位中的环烯醚萜苯丙素苷类单体化合物作为对照品TLC或HPLC检测,合并相应流份。上述流份于50℃减压浓缩蒸干,用适量蒸馏水混悬,上SephadexLH‐20柱色谱或用制备高效液相色谱技术分离纯化,依次用乙醇水由0‐20%洗脱,再用乙醇水20‐95%洗脱,洗脱液用喷雾干燥进行干燥,收集喷雾干燥粉,用95%乙醇结晶并重结晶,即得。采用上述方法制备栀子果实60%乙醇洗脱有效部位中的环烯醚萜苯丙素苷类单体化合物为0.8-5.5g/kg,经HPLC检测纯度为97.3-98.8%;色谱条件UItimate○RXB-C18色谱柱(4.6mm×150mm,3μm),流动相为乙腈-水(10%-100%梯度洗脱),检测波长为238或254nm,流速为1.0ml/min,柱温30℃,进样量20μl。
实施例5
栀子果实60%乙醇洗脱有效部位以单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分为主,其上述成分总含量大于90%,该栀子果实60%乙醇洗脱有效部位中有效成分5,7-dihydroxyl-3′,4′,5′-trimethoxyflavone、5,7,4′-trihydroxyl-6-methoxyflavone、5,7,4′-trihydroxyl-8-methoxyflavone、5,7,3′-trihydroxyl-8,4′,5′-trimethoxyflavone、5-hydroxyl-6,7,3′,4′,5′-pentamethoxyflavone、5,3′-dihydroxyl-7,4′,5′-trimethoxyflavone、5-hydroxyl-7,3′,4′,5′-tetramethoxyflavone、3,5,6,4′-tetrahydroxy-3′,5′-dimethoxyflavone、5,7,3′,4′,5′-pentame-thoxyflavone中的一种或多种黄酮类单体化合物的制备方法:将上述实例1中得到的60%乙醇洗脱部位的浸膏加入等量的100-200目硅胶拌匀,50℃减压挥干溶剂,研匀,干法上样,上常压硅胶柱(200-300目,用盐酸处理硅胶,硅胶量为浸膏的50倍),上样后依次用三氯甲烷:甲醇梯度洗脱,主要收集三氯甲烷:甲醇为20:1至9:1的洗脱流份,以上述栀子果实60%乙醇洗脱有效部位中的黄酮类单体化合物作为对照品TLC或HPLC检测,合并相应流份。上述流份于50℃减压浓缩蒸干,用适量蒸馏水混悬,上SephadexLH‐20柱色谱或用制备高效液相色谱技术分离纯化,依次用乙醇水由0‐50%洗脱,再用乙醇水50‐95%洗脱,洗脱液用喷雾干燥进行干燥,收集喷雾干燥粉,用95%乙醇结晶并重结晶,即得。采用上述方法制备栀子果实60%乙醇洗脱有效部位中的黄酮类单体化合物为0.5-7.9g/kg,经HPLC检测纯度为98.1-98.9%;色谱条件UItimate○RXB-C18色谱柱(4.6mm×150mm,3μm),流动相为乙腈-水(10%-100%梯度洗脱),检测波长为254或325nm,流速为1.0ml/min,柱温30℃,进样量20μl。
实施例6
栀子果实60%乙醇洗脱有效部位以单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分为主,其上述成分总含量大于90%,该栀子果实60%乙醇洗脱有效部位中有效成分crocin-I、crocin-II中的一种或多种藏红花素类单体化合物的制备方法:将上述实例1中得到的60%乙醇洗脱部位的浸膏加入等量的100-200目硅胶拌匀,50℃减压挥干溶剂,研匀,干法上样,上常压硅胶柱(200-300目,用盐酸处理硅胶,硅胶量为浸膏的50倍),上样后依次用三氯甲烷:甲醇梯度洗脱,主要收集三氯甲烷:甲醇为12:1至5:1的洗脱流份,以上述栀子果实60%乙醇洗脱有效部位中的藏红花素类单体化合物作为对照品TLC或HPLC检测,合并相应流份。上述流份于50℃减压浓缩蒸干,用适量蒸馏水混悬,上SephadexLH‐20柱色谱或用制备高效液相色谱技术分离纯化,依次用乙醇水由0‐50%洗脱,再用乙醇水50‐95%洗脱,洗脱液用喷雾干燥进行干燥,收集喷雾干燥粉,用95%乙醇结晶并重结晶,即得。采用上述方法制备栀子果实60%乙醇洗脱有效部位中的藏红花素类单体化合物为0.22-1.5g/kg,经HPLC检测纯度为98.1-98.9%;色谱条件UItimate○RXB-C18色谱柱(4.6mm×150mm,3μm),流动相为乙腈-水(10%-100%梯度洗脱),检测波长为440nm,流速为1.0ml/min,柱温30℃,进样量20μl。
实施例7
本发明从动物因素诱发性与AD临床病因病机相结合的异质性及多因性出发,将脑衰老及血管危险因子(高血压、动脉粥样硬化和短暂性脑缺血等)导致的脑低灌注和AD脑病理病变相结合,利用腹腔注射半乳糖促老化与反复结扎双侧颈总动脉致脑低灌注及鼠脑室一次性注射β淀粉样蛋白制作异质性及多因性阿尔茨海默病(Heterogeneitys/Multi‐factors'Alzheimer Disease,H/MAD)模型或记忆力减退或记忆障碍模型。应用异质性及多因性阿尔茨海默病模型大鼠对实施例1中水洗脱部位、30%乙醇水洗脱部位、60%乙醇水洗脱部位和京尼平苷单体进行药理活性筛选,通过行为学(Morris水迷宫)测定和应用免疫组化对模型大鼠脑内海马区(神经元)相应指标的测定与检测,筛选确定30%乙醇洗脱部位和60%乙醇洗脱部位为栀子果实药材治疗异质性及多因性阿尔茨海默病的有效部位,京尼平苷单体为栀子果实药材治疗异质性及多因性阿尔茨海默病的有效成分。栀子各有效部位和有效成分能使H/MAD模型大鼠缩短潜伏期、增加过台次数,延长在第四象限逗留时间,能使血清中总抗氧化能力T-AOC的活力恢复,降低TchE含量,增强清除自由基的能力,拮抗氧化反应,同时防止神经递质的减少,从而起到防治阿尔茨海默病(AD)或辅助改善记忆功效的药理作用。
1材料
1.1动物 SPF级健康SD雄性大鼠,体质量350-400g,由湖南斯莱克景达实验动物有限公司提供,许可证号:SCXK(湘)2009-0004。饲养条件:温度20-22℃,喂以大鼠标准饲料,自由饮水。
1.2药品与试剂 实验室自行制备栀子果实水洗脱部位、30%乙醇水洗脱部位、60%乙醇水洗脱部位、95%乙醇水洗脱部位及京尼平苷(含量98%)。盐酸多奈哌齐(卫材药业有限公司,国药准字H20050978,批号131027A);D-半乳糖(Sigma公司,批号Lot#SLBD5369V);Aβ1-40(Sigma公司,批号052M4801V);T-AOC、SOD、MDA、GSH-Px和TChE检测试剂盒(南京建成生物工程研究所,批号20141024)。
1.3仪器 ZS-B型脑立体定位仪、Morris水迷宫视频分析系统(北京众实迪创科技发展有限责任公司),Forma-86C ULT超低温冰箱(美国赛默飞),Allegra64R高速冷冻离心机(美国贝克曼库尔特),UV-1800型紫外-可见分光光度计(日本岛津公司)。
2方法
2.1药物的制备 将栀子果实150.0kg,粉碎成粗粉,用6倍量70%乙醇回流提取3次,每次2小时,合并提取液,减压浓缩提取液得浸膏19462.0g。取15731.0g浸膏水混悬,通过AB-8型大孔吸附树脂富集,依次用水、30%乙醇水、60%乙醇水、95%乙醇水洗脱,得到各洗脱部位,水洗部位为5300.3g干膏、30%乙醇水洗脱部位为3291.5g干膏、60%乙醇水洗脱部位为2163.2g干膏、95%乙醇水洗脱部位为262.5g干膏。再将30%乙醇水洗脱物经聚酰胺柱色谱纯化得总单萜类化合物(单萜类总含量大于90%)。将该部位继续分离得到单体京尼平苷(含量98%)。将上述各洗脱部位及京尼平苷用2%聚山梨酯的水溶液制备供试液。
2.2动物分组和动物模型的建立 将144只健康SD雄性大鼠随机分为空白组、假损伤组、模型组、水洗脱部位高剂量组、水洗脱部位中剂量组、30%乙醇水洗脱部位高剂量组、30%乙醇水洗脱部位中剂量组、60%乙醇水洗脱部位高剂量组、60%乙醇水洗脱部位中剂量组、京尼平苷高剂量组、京尼平苷中剂量组、盐酸多奈哌齐组,共12组,每组12只大鼠。
异质性及多因性阿尔茨海默病(AD)动物模型或记忆力减退或记忆障碍模型:各组实验大鼠每天腹腔注射D-半乳糖(50mg·kg-1),造成亚急性衰老,4W后先进行双侧颈总动脉反复结扎间段性的阻断血流,造成脑缺血性病灶(水合氯醛腹腔注射麻醉,颈正中部切口,分离双侧颈总动脉,套“4”号丝线扣。拉紧丝扣,阻断血流10min,松开丝扣使血液灌注10min后,再次阻断血流10min,第2次再灌注后观察5min,缝合皮肤。)手术完成后,将大鼠喂养3天,取未死亡的状态良好的大鼠再脑内注射凝聚态Aβ1-40(水合氯醛麻醉,固定在脑立体定位仪上,头部备皮,在无菌下操作,沿颅中线做长2~3cm的切口,分离骨膜,暴露出“人”字缝和“十”字缝。在前囟后3.0mm,中线右侧旁开2.0mm,用7号针头钻开颅骨,暴露硬脑膜,微量注射器自脑表面垂直进针4.0mm,然后向脑组织内缓慢注射5μ1Aβ1-40,注射5min,留针5min,注射后用牙科泥封堵颅骨。消炎后缝合皮肤。
空白组不予处理;模型组与给药组分别按上述造模方法予以处理;假损伤组腹腔注射生理盐水4W,4W后先进行双侧颈总动脉反复结扎(分离颈总动脉,只穿线但不结扎)再脑内注射等量的生理盐水。假损伤组与模型组于脑内注射手术后第3天开始灌胃2%吐温-80水溶液6天,每天1次。各给药组手术后第3天开始连续灌胃给药6天,每天1次。
2.3Morris水迷宫实验 预先在水池里注入清水使水面高出平台1-2cm,水温控制在24士2℃左右。水池内壁涂有黑漆,每次测试前须向水池内加入适量黑色墨水,水池共分为4个象限。将平台放在第四象限中间,在其他三个象限任选一点面向水池池壁将大鼠放入水中,试验历时5天,每天上午每个象限各测试一次,每次测试1min。若大鼠在1min之内尚未找到平台,则将大鼠拿到平台上并使它在上面站15s,若大鼠在l min之内找到平台,也让其在平台上站15s,方结束一次训练,如此训练4天。在第5天,将平台移走,每只大鼠游l min,记录每只大鼠第一次经过原平台的时间(潜伏期),每只大鼠在l min内经过平台的次数、游过的总距离以及在原平台所在象限及其它三个象限内逗留的时间,计算出在第四象限游泳的时间(t4)占整个游泳时间(t总)的百分比。
2.4分光光度法测定T-AOC、SOD、MDA、GSH-Px和TChE的含量 在2.3大鼠行为学检测后,每组大鼠用水合氯醛(300mg/kg)麻醉后,快速开胸,行心脏采血,3500r/min离心10min取血清,置于-70℃低温保存。按试剂盒说明书,紫外分光光度法检测T-AOC、SOD、MDA、GSH-Px和TChE的含量。
2.5统计方法 采用SAS软件分析,实验数据用表示,单因素方差分析实验的数据。P<0.05为有统计学意义。
3结果
3.1栀子果实不同洗脱部位及京尼平苷对H/MAD模型大鼠Morris水迷宫实验的影响
与空白组和假损伤组比较,模型组大鼠潜伏期延长、过台次减少,在第四象限逗留时间短,差异显著(P<0.01);与模型组比较,给药盐酸多奈哌齐组、30%乙醇水洗脱部位组、60%乙醇水洗脱部位组、京尼平苷组能缩短潜伏期、增加过台次数,在第四象限逗留时间延长(P<0.05或P<0.01),结果提示30%乙醇水洗脱部位组和60%乙醇水洗脱部位组及京尼平苷(尤其是60%乙醇水洗脱部位组)效果明显,起到防治阿尔茨海默病(AD)的药理作用或辅助改善记忆功效。结果见表1。
表1栀子果实不同洗脱部位及京尼平苷对H/MAD大鼠空间记忆能力的影响
注:与模型组比较,*P<0.05,**P<0.01
3.2栀子果实不同洗脱部位及京尼平苷对H/MAD模型大鼠血清中各项指标含量的影响
与空白组和假损伤组比较,模型组大鼠血清中T-AOC、SOD、GSH-PX含量明显降低(P<0.01),TChE活力增强(P<0.01),MDA水平显著升高(P<0.01);给药各组能提高T-AOC的含量,使SOD、GSH-PX明显上升(P<0.05或P<0.01),降低TChE活力,MDA水平明显下降(P<0.05或P<0.01),与模型组相比具有显著差异(P<0.05或P<0.01),说明栀子果实30%和60%乙醇水洗脱部位组及京尼平苷(尤其是60%乙醇水洗脱部位组)效果明显。能使H/MAD模型大鼠脑内T-AOC、SOD、GSH-PX的活力恢复,降低TChE、MDA含量,增强清除自由基的能力,拮抗氧化反应,同时防止神经递质的减少,起到防治阿尔茨海默病(AD)的药理作用或辅助改善记忆功效。结果见表2、表3。
表2栀子果实不同洗脱部位及京尼平苷对H/MAD模型大鼠血清中T-AOC和TChE含量的影响
注:与模型组比较,*P<0.05,**P<0.01
表3栀子果实不同洗脱部位及京尼平苷对H/MAD模型大鼠血清中SOD、MDA、GSH-PX含量影响
注:与模型组比较,*P<0.05,**P<0.01
实施例8
栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中30%乙醇洗脱有效部位以单萜苷类成分为主,其单萜苷类成分总含量大于90%,单萜苷类成分主要为京尼平苷、去乙酰车叶草苷酸甲酯、gardaloside、6-甲氧基鸡屎藤次苷甲酯、6-甲氧基去乙酰车叶草苷酸甲酯、栀子苷、ajugol、genipin-1-β-gentiobiside、genipin-1,10-di-O-β-D-glucopyranoside、gardoside methyl ester、shanzhiside、Lamalbidicacid、4-carboxyl-7-hydroxy-catapol acid、4-carboxyl- catapol acid、Mussaenosidic acid、geniposidic acid、6-hydroxymethy-jasminoside D、bornyl-6-O-β-D-xylopyranosyl-β-D-glucopyranoside、jasminoside A、epijasminoside A、jasminoside B、jasminoside E、jasminoside G、jasminoside I等单体化合物的结构确证与波谱数据信息。
8.1栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中30%乙醇洗脱有效部位中环烯醚萜苷类波谱数据信息:
化合物Z1为京尼平苷(geniposide):白色粉末(甲醇),1H-NMR(DMSO-d6,400MHz)δ:4.53(1H,d,J=7.8Hz,H-1′),7.47(1H,d,J=0.8Hz,H-3),5.68(1H,brs,H-7),4.13(1H,d,J=15.0Hz,H-10a),3.97(1H,d,J=15.0Hz,H-10b),3.64(3H,s,11-OCH3);13C-NMR(DMSO-d6,100MHz)δ:95.7(C-1),151.5(C-3),110.9(C-4),34.4(C-5),37.9(C-6),125.4(C-7),144.1(C-8),45.8(C-9),59.3(C-10),166.9(C-11),51.0(C-12),98.5(C-1′),77.2(C-3′),76.6(C-5′),73.3(C-2′),69.92(C-4′),60.9(C-6′)。
化合物Z2为去乙酰车叶草苷酸甲酯(deacetylasperulosidic acid methylester):淡黄色粉末(甲醇),1H-NMR(DMSO-d6,400MHz)δ:5.29(1H,d,J=4.8Hz,H-1),7.36(1H,d,J=0.9Hz,H-3),2.96(1H,m,H-5),4.35(1H,brs,H-6),5.65(1H,d,J=3.2Hz,H-7),2.88(1H,dd,J=8.0,3.2Hz,H-9),4.13(1H,d,J=15.2Hz,H-10a),3.97(1H,d,J=15.2Hz,H-10b),3.66(3H,s,11-OCH3),4.46(1H,d,J=8.0Hz,H-1′),3.10~3.44(4H,m,H-2′,3′,4′,5′);13C-NMR(DMSO-d6,100MHz)δ:98.4(C-1),151.5(C-3),109.3(C-4),42.7(C-5),76.6(C-6),128.8(C-7),145.7(C-8),48.6(C-9),58.8(C-10),167.3(C=O),51.1(-OCH3),94.7(C-1′),73.1(C-2′),77.2(C-3′),69.9(C-4′),77.2(C-5′),60.9(C-6′)。
化合物Z3为gardaloside:淡黄色固体(甲醇),1H-NMR(DMSO-d6,400MHz)δ:7.38(1H,brs,H-3),5.65(1H,brs,H-1),5.19(2H,brs,H-10),4.08(1H,m,H-7),3.16(1H,m,H-5),3.03(1H,m,H-9),2.64(1H,dd,J=16.8,6.8Hz,H-6a),2.36(1H,dd,J=16.8,6.8Hz,H-6b),4.74(1H,d,J=7.6Hz,H-1′),3.00~3.15(4H,m,糖上其它质子)。
化合物Z4为6-甲氧基鸡屎藤次苷甲酯(6-O-Methyl scandoside Methyl Ester):黄色透明固体(甲醇),FAB-MS m/z:419[M+H]+。1H-NMR(DMSO-d6,400MHz)δ:7.38(1H,d,J=0.9Hz,H-3),5.66(1H,brs,H-7),5.28(1H,d,J=4.8Hz,H-1),4.75(1H,brs,H-6),4.12(1H,d,J=15.3Hz,H-10a),4.00(1H,d,J=15.3Hz,H-10b),2.97(1H,m,H-5),2.87(1H,dd,J=8.0,2.4Hz,H-9),3.66(3H,s,11-COOCH3),3.17(3H,s,6-OCH3),4.47(1H,d,J=8.0Hz,H-1′),3.00~3.15 (4H,m,糖上其它质子)。
化合物Z5为6-甲氧基去乙酰车叶草苷酸甲酯(6-O-Methyldeacetylas-perulosidic Acid Methyl Ester):黄色透明固体(甲醇),FAB-MS m/z:419[M+H]+。1H-NMR(DMSO-d6,400MHz)δ:7.47(1H,d,J=0.92Hz,H-3),5.68(1H,brs,H-7),5.28(1H,d,J=6.9Hz,H-1),4.49(1H,brt,J=5.4Hz,H-6),4.13(1H,d,J=14.5Hz,H-10a),3.97(1H,d,J=14.5Hz,H-10b),3.41(1H,m,H-5),2.62(1H,dd,J=10.8,7.3Hz,H-9),3.64(3H,s,11-COOCH3),3.34(3H,s,6-OCH3),4.52(1H,d,J=8.0Hz,H-1′),3.04~3.18(4H,m,糖上其它质子)。
化合物Z6为栀子苷(gardenoside):无色针状结晶。1H-NMR(DMSO-d6,400MHz):δ:7.45(1H,s,H-3),5.62(1H,brs,H-6),4.54(1H,d,J=7.8Hz,Glc-H-1′),3.62(3H,s,11-OCH3)。
化合物Z7为ajugol:淡黄色粉末(甲醇),1H-NMR(DMSO-d6,400MHz)δ:7.10(1H,d,J=8.0Hz,H-3),6.60(1H,d,J=8.0Hz,H-1),4.75(1H,d,J=8.8Hz,H-1),4.46(1H,d,J=8.0Hz,H-1′),1.10(1H,s,CH3-10);13C-NMR(DMSO-d6,100MHz)δ:92.3(C-1),135.2(C-3),103.8(C-4),40.0(C-5),76.9(C-6),49.9(C-7),77.1(C-8),50.9(C-9),24.5(C-10),97.6(C-1′),73.3(C-2′),76.4(C-3′),69.9(C-4′),76.7(C-5′),61.0(C-6′)。
化合物Z8为京尼平-1-β-龙胆苷(genipin-1-β-gentiobiside):白色粉末(甲醇),
1H-NMR(DMSO-d6,400MHz)δ:5.08(1H,d,J=7.0Hz,H-1),7.57(1H,d,J=1.2Hz,H-3),5.88(1H,brs,H-7),4.26(1H,d,J=14.8Hz,H-10a),4.14(1H,d,J=14.8Hz,H-10b),4.94(1H,d,J=8.8Hz,H-1′),4.12(1H,dd,J=1.5,12.0Hz,H-6′),3.97(1H,dd,J=1.5,12.0Hz,H-6′),4.42(1H,d,J=8.0Hz,H-1″),3.97(1H,brs,H-6″),3.67(1H,brs,H-6″),3.65(3H,s,11-OCH3)。
化合物Z9为genipin-1,10-di-O-β-D-glucopyranoside:白色粉末(甲醇),1H-NMR(DMSO-d6,400MHz)δ:5.07(1H,d,J=7.0Hz,H-1),7.57(1H,d,J=1.2Hz,H-3),5.86(1H,brs,H-7),4.73(1H,brd,J=12.8Hz,H-10a),4.38(1H,brd,J=12.8Hz,H-10b),4.94(1H,d,J=8.8Hz,H-1′),3.97(1H,brd,J=1.5,11.2Hz,H-6′),3.65(1H,dd,J=1.5,11.2Hz,H-6′),4.54(1H,d,J=9.6Hz,H-1″),3.97(1H,brd,J=1.5,11.2Hz,H-6″),3.65(1H,dd,J=1.5,11.2Hz,H-6″),3.65(3H,s,11-OCH3)。
化合物Z10为栀子新苷甲酯(gardoside methyl ester):白色粉末(甲醇),1H-NMR(DMSO-d6,400MHz)δ:5.11(1H,d,J=6.8Hz,H-1),7.40(1H,s,H-3),5.67(brs,2H,H-10),4.37(1H,m,H-7),3.65(s,3H,-OCH3),4.42(1H,d,J=7.8Hz,H-1′);13C-NMR(DMSO-d6,100MHz,δppm):δ:95.7(C-1),153.0 (C-3),111.7(C-4),29.1(C-5),41.0(C-6),73.5(C-7),150.8(C-8),44.1(C-9),107.3(C-10),166.8(C-11),51.0(C-12),98.8(C-1′),73.4(C-2′),77.2(C-3′),69.9(C-4′),77.1(C-5′),61.0(C-6′)。
化合物Z11为山栀子苷(shanzhiside):无色针晶(甲醇),1H-NMR和13C-NMR(DMSO-d6,400MHz)δ:7.31(1H,s,H-3),5.12(1H,d,J=6.8Hz,H-1),4.73(1H,d,J=8.0Hz,H-1′),2.37(1H,m,H-9),3.67(1H,m,H-6),1.76(1H,d,J=6.5,H-7),1.64(1H,m,H-5),1.36(3H,s,H-10)。
化合物Z12为Lamalbidicacid:淡黄色粉末(甲醇),1H-NMR(DMSO-d6,400MHz)δ:7.29(1H,s,H-3),4.85(1H,d,J=8.4Hz,H-1),4.55(1H,d,J=8.0Hz,H-1′),1.23(3H,s,CH3-10);13C-NMR(DMSO-d6,100MHz)δ:96.7(C-1),148.8(C-3),128.7(C-4),41.6(C-5),75.0(C-6),76.7(C-7),92.1(C-8),45.3(C-9),22.0(C-10),173.4(C=O),98.5(C-1′),73.4(C-2′),74.6(C-3′),69.9(C-4′),76.5(C-5′),60.9(C-6′)。
化合物Z13为4-carboxyl-7-hydroxy-catapol acid:淡黄色粉末(甲醇),1H-NMR(DMSO-d6,400MHz)δ:7.23(1H,s,H-3),5.70(1H,s,H-7),4.93(1H,d,J=8.0Hz,H-1),4.53(1H,d,J=8.0Hz,H-1′);13C-NMR(DMSO-d6,100MHz)δ:96.7(C-1),145.3(C-3),128.8(C-4),44.5(C-5),80.6(C-6),128.8(C-7),149.1(C-8),45.7(C-9),59.4(C-10),169.0(C=O),98.2(C-1′),73.3(C-2′),77.1(C-3′),69.8(C-4′),76.5(C-5′),60.8(C-6′)。
化合物Z14为4-carboxyl-catapol acid:淡黄色粉末(甲醇),1H-NMR(DMSO-d6,400MHz)δ:7.37(1H,s,H-3),4.83(1H,d,J=8.4Hz,H-1),4.56(1H,d,J=8.0Hz,H-1′);13C-NMR(DMSO-d6,100MHz)δ:96.9(C-1),149.2(C-3),128.6(C-4),41.8(C-5),76.5(C-6),59.9(C-7),63.8(C-8),45.2(C-9),60.7(C-10),172.8(C=O),98.5(C-1′),73.4(C-2′),77.0(C-3′),69.6(C-4′),76.7(C-5′),61.1(C-6′)。
化合物Z15为Mussaenosidic acid:淡黄色粉末(甲醇),1H-NMR(DMSO-d6,400MHz)δ:7.31(1H,s,H-3),4.84(1H,d,J=8.4Hz,H-1),4.55(1H,d,J=8.0Hz,H-1′),1.23(1H,s,CH3-10);13C-NMR(DMSO-d6,100MHz)δ:98.5(C-1),148.9(C-3),128.6(C-4),41.7(C-5),39.2(C-6),45.2(C-7),63.7(C-8),59.7(C-9),28.9(C-10),168.4(C=O),98.7(C-1′),73.4(C-2′),76.9(C-3′),69.6(C-4′),76.5(C-5′),60.7(C-6′)。
化合物Z16为京尼平苷酸(geniposidic acid):无色针晶(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:5.34(1H,d,J=8.4Hz,H-1),7.57(1H,d,J=1.2Hz,H-3),3.38(1H,m,H-5),2.37(1H,m,H-6a),2.86(1H,m,H-6b),5.89(1H,s,H-7),2.95(1H,m,H-9),4.30(1H,d,J=12.8Hz,H-10a),4.20(1H,d,J=12.8Hz,H-10b),4.94(1H,d,J=8.4Hz,H-1′),2.97~3.56(4H,m,H-2′,3′,4′,5′)。
8.2栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中30%乙醇洗脱有效部位中单萜苷类波谱数据信息:
化合物Z17(新化合物)为6-hydroxymethy-jasminoside D:淡黄色油状。1H-NMR(DMSO-d6,400MHz)δ:1.58(1H,t,J=12.4Hz,H-2a),1.30(1H,dd,J=2.0,12.4Hz,H-2b),3.58(1H,m,H-3),3.78(1H,d,J=3.0Hz,H-4),3.90(1H,d,J=4.0Hz,H-7),1.82(3H,s,CH3-8),1.00(6H,s,CH3-9,10),4.27(1H,d,J=7.8Hz,H-1′),3.03(1H,m,H-2′),3.18(1H,m,H-3′),3.08(1H,m,H-4′),3.16(1H,m,H-5′),3.66和3.42(2H,m,H-6′);13C-NMR(DMSO-d6,100MHz)δ:36.5(C-1),42.4(C-2),65.2(C-3),83.2(C-4),141.0(C-5),129.6(C-6),56.4(C-7),17.4(C-8),26.8(C-9),28.9(C-10),104.6(C-1′),73.4(C-2′),76.8(C-3′),69.8(C-4′),76.9(C-5′),60.8(C-6′)。
化合物Z18为bornyl-6-O-β-D-xylopyranosyl-β-D-glucopyranoside:无色固体。1H-NMR(DMSO-d6,400MHz)δ:4.57(1H,d,J=7.8Hz,H-1″),4.38(1H,d,J=8.0Hz,H-1′),4.27(1H,brs,H-2),3.89(1H,dd,J=12.0,2.0Hz,H-6′a),3.80(1H,dd,J=11.6,5.6Hz,H-5″b),3.64(1H,ddd,J=10.6,9.2,5.5Hz,H-4″),3.48(1H,ddd,J=9.5,5.5,2.0Hz,H-5′),3.43(1H,t,J=9.2Hz,H-3″),3.40(1H,dd,J=9.2,9.2Hz,H-3′),3.36(1H,dd,J=9.5,9.2Hz,H-4′),3.20(1H,dd,J=11.6,10.6Hz,H-5″a),3.18(1H,dd,J=9.2,7.6Hz,H-2″),3.13(1H,dd,J=9.2,8.1Hz,H-2′),3.05(1H,m,H-6a),2.96(1H,m,H-3b),1.65(1H,m,H-5b),1.63(3H,s,H-10),1.56(1H,m,H-4),1.28(1H,m,H-6b),1.25(1H,m,H-5a),1.17(1H,m,H-3a),1.14(3H,s,H-8),0.98(3H,s,H-9);13C-NMR(DMSO-d6,100MHz)δ:48.6(C-1),76.5(C-2),38.5(C-3),40.1(C-4),39.3(C-5),39.1(C-6),34.8(C-7),28.1(C-8),29.8(C-9),19.2(C-10),100.3(C-1′),74.1(C-2′),76.8(C-3′),70.0(C-4′),76.5(C-5′),67.0(C-6′),104.2(C-1″),74.0(C-2″),76.5(C-3″),69.9(C-4″),63.0(C-5″)。
化合物Z19为jasminoside A:淡黄色透明固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:5.80(1H,brs,H-4),4.50(1H,dd,J=10.4,3.2Hz,H-7a),4.09(1H,dd,J=10.4,3.2Hz,H-7b),2.68(1H,d,J=16.7Hz,H-2a),1.85(1H,d,J=16.7Hz,H-2b),2.13(1H,brt,J=3.6Hz,H-6),2.00(3H,d,J=0.96Hz,H-10),1.05(3H,s,H-9),0.96(3H,s,H-8),4.12(1H,d,J=8.0Hz,H-1′),2.89(1H,m,H-2′),3.12(1H,m,H-3′),3.08(1H,m,H-4′),3.00(1H,m,H-5′),3.65和3.42 (2H,m,H-6′)。
化合物Z20为epijasminoside A:淡黄色透明固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:5.79(1H,brs,H-4),4.47(1H,dd,J=10.5,4.8Hz,H-7a),3.93(1H,dd,J=10.5,4.8Hz,H-7b),2.48(1H,d,J=16.5Hz,H-2a),1.91(1H,d,J=16.5Hz,H-2b),2.19(1H,brt,J=4.4Hz,H-6),2.02(3H,d,J=0.92Hz,H-10),1.03(3H,s,H-9),0.97(3H,s,H-8),4.13(1H,d,J=7.8Hz,H-1′),2.92(1H,m,H-2′),3.12(1H,m,H-3′),3.10(1H,m,H-4′),3.05(1H,m,H-5′),3.74和3.44(2H,m,H-6′)。
化合物Z21为jasminoside B:淡黄色油状。1H-NMR(DMSO-d6,400MHz)δ:6.17(1H,s,H-4),4.59(1H,dd,J=17.0,1.50Hz,H-10a),4.16(1H,dd,J=17.0,1.50Hz,H-10b),3.70(2H,m,H-7),2.62(1H,d,J=16.8Hz,H-2a),1.91(1H,d,J=16.8Hz,H-2b),2.00(1H,t,J=4.0Hz,H-6),1.06(3H,s,H-8),0.95(3H,s,H-9),4.20(1H,d,J=7.8Hz,H-1′),3.03(1H,m,H-2′),3.41(1H,m,H-3′),3.09(1H,m,H-4′),3.45(1H,m,H-5′),3.64和4.14(2H,m,H-6′);13C-NMR(DMSO-d6,100MHz)δ:35.0(C-1),48.7(C-2),198.6(C-3),122.9(C-4),161.7(C-5),48.8(C-6),60.5(C-7),26.7(C-8),28.7(C-9),69.3(C-10),102.7(C-1′),73.5(C-2′),76.6(C-3′),70.0(C-4′),77.0(C-5′),61.0(C-6′)。
化合物Z22为jasminoside E:淡黄色冻状固体。1H-NMR(DMSO-d6,400MHz)δ:4.48(1H,d,J=11.2,H-7a),4.00(1H,d,J=11.2,H-7b),2.40(1H,dd,J=17.2,5.6Hz,H-3a),1.92(1H,dd,J=17.2,9.2Hz,H-3b),1.78(1H,dd,J=12.4,5.6Hz,H-2a),1.36(1H,t,J=12.4Hz,H-2b),1.65(3H,s,H-10),1.15(3H,s,H-8),1.02(3H,s,H-9),4.28(1H,d,J=8.0Hz,H-1′),2.92(1H,m,H-2′),3.15(1H,m,H-3′),3.10(1H,m,H-4′),3.02(1H,m,H-5′),3.67和3.45(2H,m,H-6′);13C-NMR(DMSO-d6,100MHz)δ:35.0(C-1),44.9(C-2),37.2(C-3),171.1(C-4),129.0(C-5),135.4(C-6),70.0(C-7),28.4(C-8),29.0(C-9),20.8(C-10),100.8(C-1′),73.5(C-2′),76.7(C-3′),70.0(C-4′),76.8(C-5′),61.0(C-6′)。
化合物Z23为jasminoside G:淡黄色油状。1H-NMR(DMSO-d6,400MHz)δ:6.15(1H,s,H-4),4.37(1H,dd,J=17.6,1.40Hz,H-10a),4.12(1H,dd,J=17.6,1.40Hz,H-10b),4.05(1H,dd,J=16.0,6.4Hz,H-7a),3.88(1H,dd,J=16.0,6.4Hz,H-7b),2.62(1H,d,J=16.8Hz,H-2a),1.89(1H,d,J=16.8Hz,H-2b),2.19(1H,t,J=4.3Hz,H-6),1.06(3H,s,H-8),0.93(3H,s,H-9),4.20(1H,d,J=8.0Hz,H-1′),3.09(1H,m,H-2′),3.42(1H,m,H-3′),3.13(1H,m,H-4′),3.44 (1H,m,H-5′),3.65和4.13(2H,m,H-6′)。
化合物Z24为jasminoside I:黄色透明固体(甲醇),1H-NMR(DMSO-d6,400MHz)δ:4.96(1H,d,J=10.4Hz,H-7a),4.82(1H,d,J=10.4Hz,H-7b),1.96(2H,brt,J=6.2Hz,H-3a,3b),1.60(2H,m,H-2a,2b),1.63(3H,s,H-10),1.07(3H,s,H-8),1.05(3H,s,H-9),5.53(1H,d,J=8.2Hz,H-1″),4.20(1H,d,J=7.5Hz,H-1′),3.57~3.68(2H,m,H-6′,6″),2.92~3.44(m,糖上其它质子)。
实施例9
栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中60%乙醇洗脱有效部位以单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分为主,其上述成分总含量大于90%。其中,单萜类苷元成分主要为jasminoside C aglycone、5,5-dihydroxymethyl-1-cyclohexene-furan lactone、6-hydroxyl-3-hydroxy-methyl-4,4-dimethyl-1-cyclohexene-1-formaldehyde、4-hydro-xyl-2-hydroxymethyl-6,6-dimethyl-1-cyclohexene-1-formaldehyde、4-hydroxyl-1,5,5-trimethyl-6-epoxyethyl-cyclohex-3-enone、iridoids 2a、iridoids 2b、jasminodiol、crocusatin-C、7-Epi-crocusatin-C、(10S,11S)-Gardendiol、(10R,11R)-Gardendiol、(5S,9S)-Gardenate A、(5R,9R)-Gardenate A、5,6-dihydroxymethyl-1,1-dimethylcyclo-hex-4-enone、1-hydroxy-7-hydroxylmethyl-1,4a,5,7a-tetrahydro-pyrancyclopenta-4-carbaldehyde或环烯醚萜苯丙素苷类成分主要为5-aldehyde-6′-O-sinapoyljasmino-side A、6′-O-sinapoyljasminoside B、6′-O-trans-sinapoyljasminoside L、6′-O-sinapoyl-3-methoxy-4-deketone-jasminoside E、6″-O-trans-sina-poylgenipingentiobioside、6″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin、6″-O-[3″′-methoxy-(E)-caffeoyl]-gentiobiosylgenipin、4″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin、6″-O-[(E)-feruloyl]-gentiobiosylgenipin、6″-O-[(E)-cinnamoyl]-gentiobiosylgenipin或黄酮类成分主要为5,7-dihydroxyl-3′,4′,5′-trimethoxyflavone、5,7,4′-trihydroxyl-6-methoxyflavone、5,7,4′-trihydroxyl-8-methoxyflavone、5,7,3′-trihydroxyl-8,4′,5′-trimethoxyflavone、5-hydroxyl-6,7,3′,4′,5′-pentamethoxyflavone、5,3′-dihydroxyl-7,4′,5′-trimethoxyflavone、5-hydroxyl-7,3′,4′,5′-tetramethoxyflavone、3,5,6,4′-tetrahydroxy-3′,5′-dimethoxyflavone、5,7,3′,4′,5′-pentame-thoxyflavone或二萜类成分主要为crocin-I、crocin-II等单体化合物的结构确证与波谱数据信息。
9.1栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中60%乙醇洗脱有效部位中单萜类苷元波谱数据信息:
化合物Z25(新化合物)为jasminoside C aglycone(1,1-dimethyl-5-hydroxymethyl-6-vinyl-cyclohex-3-enone):淡黄色固体(甲醇)。1H-NMR(MeOD,400MHz)δ:6.20(1H,d,J=1.2Hz,H-4),5.43(1H,dd,J=1.2,14.4Hz,H-7),4.43(1H,d,J=1.2Hz,H-10),2.38(1H,brs,H-2),1.20(6H,brs,CH3-8,9);13C-NMR(MeOD,100MHz)δ:39.9(C-1),53.0(C-2),201.9(C-3),123.8(C-4),158.8(C-5),150.2(C-6),113.9(C-7),28.5(C-8),28.5(C-9),61.9(C-10)。
化合物Z26(新化合物)为5,5-dihydroxymethyl-1-cyclohexene-furan lactone:淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:5.59(1H,d,J=1.2Hz,H-2),4.37(1H,dd,J=5.2,10.8Hz,H-9a),4.03(1H,t,J=10.8Hz,H-9b),3.93(1H,d,J=2.8Hz,H-11),3.62(1H,t,J=4.8Hz,H-10),3.00(1H,m,H-6),2.67(1H,m,H-3a),2.62(1H,m,H-4),2.25(1H,m,H-3b);13C-NMR(DMSO-d6,100MHz)δ:142.2(C-1),126.1(C-2),38.5(C-3),46.2(C-4),35.4(C-5),45.3(C-6),173.6(C-7),67.7(C-9),59.2(C-10),58.7(C-11)。
化合物Z27(新化合物)为6-hydroxyl-3-hydroxymethyl-4,4-dimethyl-1-cyclohex-ene-1-formaldehyde:淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:6.52(1H,d,J=2.8Hz,H-2),4.47(1H,d,J=8.8Hz,H-8a),4.27(1H,brs,H-6),3.94(1H,d,J=8.8Hz,H-8b),2.95(1H,m,H-3),1.74(1H,dd,J=5.6,12.8Hz,H-5a),1.34(1H,dd,J=10.0,12.8Hz,H-5a),0.98(3H,s,CH3-10),0.78(3H,s,CH3-9);13C-NMR(DMSO-d6,100MHz)δ:128.8(C-1),136.0(C-2),45.5(C-3),34.6(C-4),46.2(C-5),64.9(C-6),169.5(C-7),67.9(C-8),19.4(C-9),29.0(C-10)。
化合物Z28(新化合物)为4-hydroxyl-2-hydroxymethyl-6,6-dimethyl-1-cyclohex-ene-1-formaldehyde:淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:4.85(1H,t,J=2.4Hz,H-8a),4.83(1H,dd,J=2.9,16.0Hz,H-8b),3.91(1H,d,J=9.2Hz,H-4),2.38(1H,m,H-3a),1.84(1H,m,H-3b),1.72(1H,m,H-5a),1.46(1H,m,H-5b),1.15(3H,s,CH3-9),1.13(3H,s,CH3-10);13C-NMR(DMSO-d6,100MHz)δ:168.6(C-1),121.5(C-2),29.6(C-3),63.4(C-4),46.4(C-5),33.7(C-6),173.4(C-7),69.2(C-8),27.8(C-9),27.8(C-10)。
化合物Z29(新化合物)为4-hydroxyl-1,5,5-trimethyl-6-epoxyethyl-cyclohex-3-enone:淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:5.88(1H,t,J=1.6Hz,H-2),3.85(1H,d,J=7.2Hz,H-7a),3.75(1H,d,J=1.6Hz,H-4),3.45(1H,d,J=7.2Hz,H-7b),2.00(1H,s,H-8),1.08(3H,s,CH3-10),0.85(3H,s,CH3-9);13C-NMR(DMSO-d6,100MHz)δ:169.6(C-1),123.5(C-2),195.2(C-3),90.4(C-4),49.7(C-5),82.4(C-6),72.7(C-7),19.0(C-8),15.9(C-9),21.1(C-10)。
化合物Z30为iridoids 2a:淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:5.72(1H,m,H-6),4.37(1H,m,H-3),4.08(1H,m,H-9a),4.04(1H,m,H-9b),3.66(1H,m,H-7a),3.65(3H,s,CH3-10),3.07(1H,m,H-4a),2.86(1H,m,H-4),2.67(1H,m,H-5a),2.21(1H,m,H-5b);13C-NMR(DMSO-d6,100MHz)δ:171.7(C-1),66.4(C-3),44.7(C-4),37.2(C-4a),37.6(C-5),126.6(C-6),141.5(C-7),48.2(C-7a),170.9(C-8),59.1(C-9),52.0(C-10)。
化合物Z30为iridoids 2b:淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:5.73(1H,m,H-6),4.32(1H,m,H-3),3.97(2H,m,H-9),3.66(3H,s,CH3-10),3.64(1H,m,H-7a),3.36(1H,m,H-4),3.17(1H,m,H-4a),2.46(1H,m,H-5a),2.16(1H,m,H-5b);13C-NMR(DMSO-d6,100MHz)δ:170.8(C-1),64.3(C-3),41.2(C-4),35.7(C-4a),33.4(C-5),127.1(C-6),141.0(C-7),51.8(C-7a),169.3(C-8),58.5(C-9),50.1(C-10)。
化合物Z31为jasminodiol:淡黄色油状,EI-MS m/z:184[M]+。1H-NMR(DMSO-d6,400MHz)δ:5.97(1H,s,H-4),4.27(1H,dd,J=17.0,1.60Hz,H-10a),4.02(1H,dd,J=17.0,1.60Hz,H-10b),3.66(2H,m,H-7),2.61(1H,d,J=16.8Hz,H-2a),1.88(1H,d,J=16.8Hz,H-2b),1.94(1H,t,J=4.0Hz,H-6),1.06(3H,s,H-8),0.93(3H,s,H-9);13C-NMR(DMSO-d6,100MHz)δ:35.0(C-1),48.7(C-2),198.5(C-3),121.7(C-4),166.5(C-5),48.8(C-6),60.7(C-7),26.8(C-8),28.6(C-9),63.0(C-10)。
化合物Z32为crocusatin-C:淡黄色固体,1H-NMR(DMSO-d6,400MHz)δ:5.79(1H,s,H-4),3.61和3.50(2H,m,H-7),2.82(1H,d,J=16.8Hz,H-2a),2.01(1H,d,J=16.8Hz,H-2b),2.49(1H,m,H-6),1.92(3H,d,J=1.60Hz,H-10),1.01(3H,s,H-8),0.90(3H,s,H-9);13C-NMR(DMSO-d6,100MHz)δ:38.9(C-1),49.7(C-2),197.8(C-3),126.6(C-4),164.4(C-5),45.5(C-6),64.5(C-7),23.5(C-8),24.9(C-9),19.5(C-10)。
化合物Z33为7-Epi-crocusatin-C即(7S)-6-羟甲基-1,1,5-三甲基环己-3-烯酮((7S)-6-(hydroxymethyl)-1,1,5-trimethylcyclohex-3-enone):淡黄色固体。1H-NMR(DMSO-d6,400MHz)δ:5.80(1H,s,H-4),3.70(2H,d,J=4.0Hz,H-7a,7b),2.56(1H,d,J=16.8Hz,H-2a),1.84(1H,d,J=16.8Hz,H-2b),1.98(3H,d,J=0.8Hz,H-10),1.93(1H,t,J=4.0Hz,H-6),1.06(3H,s,H-8),0.94(3H,s,H-9);13C-NMR(DMSO-d6,100MHz)δ:34.9(C-1),48.3(C-2),198.5(C-3),126.0(C-4),162.5(C-5),52.8(C-6),59.9(C-7),26.7(C-8),28.8(C-9),23.6(C-10)。
化合物Z34为(10S,11S)-栀子二醇(10S,11S)-Gardendiol:淡黄色固体。1H-NMR(DMSO-d6,400MHz)δ:5.71(1H,m,H-7),4.42(1H,dd,J=10.8,5.2Hz,H-1a),4.02(1H,t,J=10.8Hz,H-1b),3.57(2H,m,H-10a,10b),3.38(2H,m,H-11a,11b),3.32(1H,m,H-9),2.32(1H,dd,J=16.8,9.0Hz,H-6a),2.07(1H,d,J=16.8Hz,H-6b),1.57(1H,m,H-4),1.40(1H,m,H-5);13C-NMR(DMSO-d6,100MHz)δ:66.5(C-1),172.9(C-3),34.5(C-4),34.2(C-5),37.0(C-6),127.4(C-7),143.6(C-8),39.0(C-9),59.5(C-10),59.1(C-11)。
化合物Z35为(10R,11R)-栀子二醇(10R,11R)-Gardendiol):淡黄色固体。1H-NMR(DMSO-d6,400MHz)δ:5.72(1H,m,H-7),4.22(2H,m,H-1a,1b),4.03(2H,m,H-10a,10b),3.56(2H,m,H-11a,11b),2.84(1H,m,H-9),2.62(1H,dd,J=16.8,9.0Hz,H-6a),2.08(1H,d,J=16.8Hz,H-6b),2.02(1H,m,H-4),1.52(1H,m,H-5);13C-NMR(DMSO-d6,100MHz)δ:66.5(C-1),171.4(C-3),29.3(C-4),29.2(C-5),38.2(C-6),126.7(C-7),141.6(C-8),34.5(C-9),59.2(C-10),49.5(C-11)。
化合物Z36为(5S,9S)-Gardenate A或(5R,9R)-Gardenate A:淡黄色固体。1H-NMR(DMSO-d6,400MHz)δ:5.73(1H,d,J=1.6Hz,H-7),4.05(2H,d,J=16.4Hz,H-10),3.98(1H,dd,J=14.8,3.6Hz,H-3b),3.88(1H,dd,J=14.8,7.2Hz,H-3a),3.64(3H,s,11位-COOCH3质子),3.51(3H,s,1位-COOCH3质子),3.50(1H,m,H-9),2.87(1H,m,H-5),2.66(1H,ddd,J=14.8,7.2,3.6Hz,H-4),2.48(1H,m,H-6a),2.38(1H,m,H-6b);13C-NMR(DMSO-d6,100MHz)δ:170.8(C-1),64.3(C-3),49.4(C-4),41.4(C-5),37.2(C-6),127.1(C-7),141.5(C-8),51.1(C-9),59.1(C-10),172.6(C-11),51.9/51.1(COOCH3);(5R,9R)-Gardenate A:淡黄色固体。1H-NMR(DMSO-d6,400MHz)δ:5.73(1H,d,J=1.6Hz,H-7),4.81(2H,brs,H-10),4.36(1H,dd,J=11.2,3.8Hz,H-3b),4.30(1H,dd,J=11.2,7.0Hz,H-3a),3.67(3H,s,11位-COOCH3质子),3.57(3H,s,1位-COOCH3质子),3.68(1H,m,H-9),3.17(1H,m,H-5),3.05(1H,ddd,J=11.2,7.0,3.8Hz,H-4),2.65(1H,m,H-6a),2.56(1H,m,H-6b);13C-NMR(DMSO-d6,100MHz)δ:172.6(C-1),66.4(C-3),58.5(C-4),48.2(C-5),37.6(C-6),127.1(C-7),143.7(C-8),59.1(C-9),61.9(C-10),173.9(C-11),52.0/51.8(COOCH3)。
化合物Z37为5,6-二羟甲基-1,1-二甲基环己-4-烯酮(5,6-dihydroxymethyl-1,1-di-methylcyclohex-4-enone):淡黄色固体。1H-NMR(DMSO-d6,400MHz)δ:4.90(1H,d,J=10.8Hz,H-7a),4.83(1H,d,J=10.8Hz,H-7b),4.86(1H,d,J=10.4Hz,H-8a),3.90(1H,d,J=10.4Hz,H-8b),2.39(1H,dd,J=15.6,4.0Hz,H-3a),1.83(1H,dd,J=15.6,4.0Hz,H-3b),1.73(1H,dd,J=10.8,8.8Hz,H-2a),1.46(1H,dd,J=10.8,4.0Hz,H-2b),1.15(3H,s,H-9),1.13(3H,s,H-10);13C-NMR(DMSO-d6,100MHz)δ:33.7(C-1),46.4(C-2),29.6(C-3),173.4(C-4),121.5(C-5),168.4(C-6),69.2(C-7),63.4(C-8),27.8(C-9),27.8(C-10)。
化合物Z38为1-羟基-7-羟甲基-1,4a,5,7a-四氢化环戊二烯骈吡喃-4-醛(1-hydroxy-7-hydroxylmethyl-1,4a,5,7a-tetrahydro-pyrancyclopenta-4-carbaldehyde):白色粉末(甲醇),1H-NMR(DMSO-d6,400MHz)δ:9.84(1H,d,J=4.4Hz,H-11),7.35(1H,d,J=5.2Hz,H-3),5.71(1H,s,H-7),4.79(1H,brs,H-1),4.31(1H,d,J=15.0,H-10a),4.14(1H,d,J=15.0,H-10b)。
9.2栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中60%乙醇洗脱有效部位中环烯醚萜苯丙素苷类波谱数据信息:
化合物Z39(新化合物)为5-aldehyde-6′-O-sinapoyljasminoside A:淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:2.60(1H,d,J=16.0Hz,H-2a),1.87(1H,d,J=16.0Hz,H-2b),6.16(1H,brs,H-4),1.93(1H,t,J=4.0Hz,H-5),4.51(1H,dd,J=1.2,16.8Hz,H-7a),4.18(1H,d,J=16.8Hz,H-7b),8.99(1H,brs,HOC-4),0.92(3H,s,CH3-9),1.00(3H,s,CH3-10),4.27(1H,d,J=7.6Hz,Glc-H-1′),3.07(1H,m,H-2′),3.19(1H,m,H-3′),3.17(1H,m,H-4′),3.41(1H,m,H-5′),4.37和4.17(2H,m,H-6′)6.56(1H,d,J=16.0Hz,H-2″),7.56(1H,d,J=16.0Hz,H-3″),7.05(2H,s,H-5″,9″),3.81(6H,s,6″,8″-2×OCH3);13C-NMR(DMSO-d6,100MHz)δ:34.9(C-1),48.8(C-2),198.6(C-3),123.1(C-4),167.0(C-5),48.5(C-6),69.6(C-7),161.6(C-8),28.3(C-9),26.6(C-10),56.0(6″,8″-OCH3),102.8(C-1′),73.5(C-2′),76.3(C-3′),69.9(C-4′),73.8(C-5′),63.4(C-6′),166.6(C-1″),114.7(C-2″),145.5(C-3″),124.3(C-4″),106.2(C-5″),148.0(C-6″),138.3(C-7″),148.0(C-8″),106.2(C-9″),56.0(6″,8″-OCH3)。
化合物Z40(新化合物)为6′-O-sinapoyljasminoside B:淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:2.71(1H,d,J=16.8Hz,H-2a),1.98(1H,d,J=16.8Hz,H-2b),6.24(1H,brs,H-4),2.09(1H,t,J=4.0Hz,H-6),4.57(1H,dd,J=0.8,12.0Hz,H-7a),4.36(1H,d,J=12.0Hz,H-7b),4.51(1H,dd,J=2.0,12.0Hz,H-7a),4.30(1H,d,J=12.0Hz,H-7b),0.99(3H,s,CH3-9),1.09(3H,s,CH3-10),4.37(1H,d,J=7.6Hz,Glc-H-1′),3.26(1H,m,H-2′),3.51(1H,m,H-3′),3.36(1H,m,H-4′),3.38(1H,m,H-5′),4.57和4.33(2H,m,H-6′),6.42(1H,d,J=16.0Hz,H-2″),7.61(1H,d,J=16.0Hz,H-3″),6.94(2H,s,H-5″,9″),3.86(6H,s,6″,8″-2×OCH3);13C-NMR(DMSO-d6,100MHz)δ:36.3(C-1),50.0(C-2),202.8(C-3),125.5(C-4),164.0(C-5),50.5(C-6),72.0(C-7),64.6(C-8),29.3(C-9),27.4(C-10),56.9(6″,8″-OCH3),104.6(C-1′),75.1(C-2′),75.6(C-3′),71.7(C-4′),77.9(C-5′),62.1(C-6′),169.1(C-1″),115.8(C-2″),147.4(C-3″),126.6(C-4″),107.0(C-5″),149.5 (C-6″),139.7(C-7″),149.5(C-8″),107.0(C-9″),56.9(6″,8″-OCH3)。
化合物Z41(新化合物)为6′-O-trans-sinapoyljasminoside L:淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:1.90(1H,dd,J=2.0,12.0Hz,H-2a),1.46(1H,t,J=12.0Hz,H-2b),4.05(1H,m,H-3),2.42(1H,dd,J=5.6,17.2Hz,H-4a),2.05(1H,dd,J=9.2,17.2Hz,H-4b),1.69(3H,s,CH3-8),0.98(3H,s,CH3-9),1.18(3H,s,CH3-10),4.43(1H,d,J=8.0Hz,Glc-H-1′),3.21(1H,m,H-2′),3.40(1H,m,H-3′),3.34(1H,m,H-4′),3.56(1H,m,H-5′),4.49和4.32(2H,m,H-6′),6.38(1H,d,J=16.0Hz,H-2″),7.60(1H,d,J=16.0Hz,H-3″),6.85(2H,s,H-5″,9″),3.85(6H,s,6″,8″-2×OCH3);13C-NMR(DMSO-d6,100MHz)δ:36.5(C-1),46.6(C-2),74.0(C-3),38.6(C-4),131.4(C-5),137.0(C-6),169.4(C-7),21.4(C-8),29.8(C-9),29.3(C-10),56.9(6″,8″-OCH3),103.4(C-1′),75.1(C-2′),78.0(C-3′),72.1(C-4′),75.4(C-5′),65.0(C-6′),168.9(C-1″),115.8(C-2″),147.3(C-3″),126.5(C-4″),106.9(C-5″),149.4(C-6″),139.6(C-7″),149.4(C-8″),106.9(C-9″),56.9(6″,8″-OCH3)。
化合物Z42(新化合物)为6′-O-sinapoyl-3-methoxy-4-deketone-jasminosideE:淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:1.78(1H,d,J=11.6Hz,H-2a),1.33(1H,d,J=11.6Hz,H-2b),3.93(1H,m,H-3),2.35(1H,d,J=6.8Hz,H-4a),1.90(1H,d,J=9.6Hz,H-4b),4.36(1H,d,J=10.4Hz,H-7a),4.03(1H,d,J=10.4Hz,H-7b),1.61(3H,s,CH3-8),0.94(3H,s,CH3-9),1.11(3H,s,CH3-10),3.34(3H,s,CH3O-3),4.32(1H,d,J=7.6Hz,Glc-H-1′),3.08(1H,m,H-2′),3.42(1H,m,H-3′),3.18(1H,m,H-4′),3.35(1H,m,H-5′),4.23和3.93(2H,m,H-6′),6.50(1H,d,J=16.0Hz,H-2″),7.54(1H,d,J=16.0Hz,H-3″),6.98(2H,s,H-5″,9″),3.79(6H,s,6″,8″-2×OCH3);13C-NMR(DMSO-d6,100MHz)δ:35.0(C-1),38.0(C-2),67.3(C-3),45.1(C-4),128.6(C-5),131.5(C-6),73.3(C-7),20.7(C-8),28.3(C-9),28.9(C-10),55.7(C-11),55.9(6″,8″-OCH3),104.0(C-1′),73.5(C-2′),76.5(C-3′),70.2(C-4′),73.5(C-5′),63.6(C-6′),166.4(C-1″),114.6(C-2″),145.3(C-3″),124.2(C-4″),106.0(C-5″),147.9(C-6″),138.2(C-7″),147.9(C-8″),106.0(C-9″),55.9(6″,8″-OCH3)。
化合物Z43为6″-O-反式-芥子酰基-京尼平龙胆二糖苷(6″-O-trans-sinapoylgenipin gentiobioside):淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:5.09(1H,d,J=4.8Hz,H-1),7.47(1H,brs,H-3),3.03(1H,m,H-5),2.67(1H,m,H-6a),2.13(1H,m,H-6b),5.72(1H,brs,H-7),2.60(1H,m,H-9),3.92(1H,d,J=11.2Hz,H-10a),3.57(1H,d,J=11.2Hz,H-10b),3.64(3H,s,11-OCH3),4.54(1H,d,J=7.6Hz,Glc-H-1′),4.14(1H,d,J=12.4Hz,Glc-H-6′a),4.00(1H,d,J=12.4Hz,Glc-H-6′b),4.28(1H,d,J=7.6Hz,Glc-H-1″),4.36(1H,d,J=10.6Hz,Glc-H-6″a),4.16(1H,d,J=10.6Hz,Glc-H-6″b),6.57(1H,d,J=16.0Hz,H-2″′),7.57(1H,d,J=16.0Hz,H-3″′),7.05(2H,s,H-5″′,9″′),3.81(6H,s,6″′,8″′-2×OCH3);13C-NMR(DMSO-d6,100MHz)δ:96.6(C-1),151.6(C-3),110.6(C-4),34.8(C-5),38.0(C-6),125.9(C-7),144.0(C-8),45.5(C-9),68.2(C-10),166.9(C-11),51.0(11-OCH3),99.0(C-1′),70.2(C-2′),76.5(C-3′),73.5(C-4′),76.2(C-5′),59.4(C-6′),103.3(C-1″),69.8(C-2″),76.5(C-3″),73.4(C-4″),73.1(C-5″),63.5(C-6″),166.7(C-1″′),114.7(C-2″′),145.6(C-3″′),124.3(C-4″′),106.2(C-5″′,9″′),148.0(C-6″′,8″′),138.2(C-7″′),56.1(6″′,8″′-OCH3)。
化合物Z44为6″-O-[(E)-p-香豆酰基]京尼平龙胆二糖苷(6″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin):淡黄色固体(甲醇)。1H-NMR(MeOD,400MHz)δ:5.12(1H,d,J=7.6Hz,H-1),7.46(1H,brs,H-3),3.12(1H,m,H-5),2.77(1H,m,H-6a),2.11(1H,m,H-6b),5.82(1H,brs,H-7),2.68(1H,m,H-9),3.68(3H,s,11-OCH3),4.70(1H,d,J=8.0Hz,Glc-H-1′),4.30(1H,d,J=14.4Hz,Glc-H-6′a),4.18(1H,d,J=14.4Hz,Glc-H-6′b),4.39(1H,d,J=7.6Hz,Glc-H-1″),4.50(1H,dd,J=12.0,1.6Hz,Glc-H-6″a),4.27(1H,dd,J=12.0,1.6Hz,Glc-H-6″b),6.35(1H,d,J=16.0Hz,H-2″′),7.62(1H,d,J=16.0Hz,H-3″′),7.45(2H,d,J=8.4Hz,H-5″′,9″′),6.80(2H,d,J=8.4Hz,H-6″′,8″′);13C-NMR(MeOD,100MHz)δ:98.9(C-1),153.5(C-3),112.3(C-4),36.7(C-5),39.8(C-6),127.2(C-7),144.8(C-8),46.9(C-9),61.6(C-10),169.6(C-11),51.9(11-OCH3),100.6(C-1′),74.8(C-2′),77.8(C-3′),71.7(C-4′),77.6(C-5′),70.1(C-6′),105.0(C-1″),75.1(C-2″),77.8(C-3″),71.6(C-4″),75.4(C-5″),64.7(C-6″),169.3(C-1″′),115.0(C-2″′),147.0(C-3″′),129.1(C-4″′),116.9(C-5″′,9″′),131.4(C-6″′,8″′),161.3(C-7″′)。
化合物Z45为6″-O-[3″′-甲氧基-(E)-咖啡酰氧基]京尼平龙胆二糖苷(6″-O-[3″′-methoxy-(E)-caffeoyl]-gentiobiosylgenipin):淡黄色固体(甲醇)。1H-NMR(MeOD,400MHz)δ:5.13(1H,d,J=8.0Hz,H-1),7.46(1H,brs,H-3),3.15(1H,m,H-5),2.80(1H,m,H-6a),2.15(1H,m,H-6b),5.81(1H,brs,H-7),2.70(1H,m,H-9),3.70(3H,s,11-OCH3),4.70(1H,d,J=8.0Hz,Glc-H-1′),4.29(1H,d,J=14.4Hz,Glc-H-6′a),4.18(1H,d,J=14.4Hz,Glc-H-6′b),4.39(1H,d,J=7.6Hz,Glc-H-1″),4.50(1H,dd,J=12.0,1.6Hz,Glc-H-6″a),4.27(1H,dd,J=12.0,1.6Hz,Glc-H-6″b),6.31(1H,d,J=16.0Hz,H-2″′),7.60(1H,d,J=16.0Hz,H-3″′),7.48(1H,dd,J=1.2,8.8Hz,H-5″′),7.43(1H,d,J=1.2Hz,9″′),6.79(1H,d,J=8.8Hz,H-8″′);13C-NMR(MeOD,100MHz)δ:98.8(C-1),153.4(C-3),112.4(C-4),36.8(C-5),39.8(C-6),127.1(C-7),144.8(C-8),47.0(C-9),61.5(C-10),169.6(C-11),51.8(11-OCH3),100.6(C-1′),74.8(C-2′),78.0(C-3′),71.7(C-4′),77.9(C-5′),69.8(C-6′),104.8(C-1″),75.2(C-2″),77.8(C-3″),71.6(C-4″),75.4(C-5″),62.8(C-6″),169.6(C-1″′),116.9(C-2″′),146.9(C-3″′),131.2(C-4″′),114.9(C-5″′),146.6(C-6″′),161.4(C-7″′),116.0(C-8″′),129.0(C-9″′)。
化合物Z46为4″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin:淡黄色固体(甲醇)。1H-NMR(MeOD,400MHz)δ:5.14(1H,d,J=7.6Hz,H-1),7.46(1H,brs,H-3),3.16(1H,m,H-5),2.80(1H,m,H-6a),2.15(1H,m,H-6b),5.81(1H,brs,H-7),2.69(1H,m,H-9),3.69(3H,s,11-OCH3),4.70(1H,d,J=8.0Hz,Glc-H-1′),4.35(1H,dd,J=3.2,16.0Hz,Glc-H-6′a),4.28(1H,d,J=16.0Hz,Glc-H-6′b),4.38(1H,d,J=7.6Hz,Glc-H-1″),4.48(1H,dd,J=12.0,1.6Hz,Glc-H-6″a),4.25(1H,dd,J=12.0,1.6Hz,Glc-H-6″b),6.36(1H,d,J=16.0Hz,H-2″′),7.64(1H,d,J=16.0Hz,H-3″′),7.47(2H,d,J=8.4Hz,H-5″′,9″′),6.79(2H,d,J=8.4Hz,H-6″′,8″′);13C-NMR(MeOD,100MHz)δ:98.8(C-1),153.4(C-3),112.3(C-4),36.8(C-5),39.7(C-6),127.1(C-7),144.8(C-8),46.9(C-9),61.5(C-10),169.6(C-11),51.7(11-OCH3),100.6(C-1′),78.0(C-2′),74.8(C-3′),77.9(C-4′),71.6(C-5′),64.7(C-6′),105.0(C-1″),75.4(C-2″),77.6(C-3″),71.9(C-4″),77.8(C-5″),62.8(C-6″),169.2(C-1″′),114.9(C-2″′),146.9(C-3″′),129.0(C-4″′),131.3(C-5″′),116.8(C-6″′),161.2(C-7″′),116.8(C-8″′),131.3(C-9″′)。
化合物Z47为6″-O-[(E)-阿魏酸基]京尼平龙胆二糖苷(6″-O-[(E)-feruloyl]-gentio-biosylgenipin):淡黄色固体(甲醇)。1H-NMR(MeOD,400MHz)δ:5.14(1H,d,J=8.0Hz,H-1),7.44(1H,brs,H-3),3.16(1H,m,H-5),2.82(1H,m,H-6a),2.15(1H,m,H-6b),5.82(1H,brs,H-7),2.69(1H,m,H-9),3.69(3H,s,11-OCH3),4.70(1H,d,J=8.0Hz,Glc-H-1′),4.28(1H,d,J=12.0Hz,Glc-H-6′a),4.19(1H,d,J=12.0Hz,Glc-H-6′b),4.38(1H,d,J=7.6Hz,Glc-H-1″),4.50(1H,dd,J=2.0,12.0Hz,Glc-H-6″a),4.11(1H,dd,J=2.0,12.0Hz,Glc-H-6″b),6.36(1H,d,J=16.0Hz,H-2″′),7.64(1H,d,J=16.0Hz,H-3″′),7.50(1H,d,J=1.2Hz,5″′),7.47(1H,dd,J=1.2,8.8Hz,H-5″′),6.78(1H,d,J=8.8Hz,H-8″′);13C-NMR(MeOD,100MHz)δ:98.8(C-1),153.4(C-3),112.3(C-4),36.8(C-5),39.7(C-6),127.1(C-7),144.8(C-8),46.9(C-9),61.5(C-10),169.6(C-11),51.7(11-OCH3),100.6(C-1′),74.8(C-2′),78.0(C-3′),71.6(C-4′),77.9(C-5′),69.7(C-6′),105.0(C-1″),75.1(C-2″),77.8(C-3″),71.8(C-4″),75.4(C-5″),64.8(C-6″),169.2(C-1″′),116.9(C-2″′),146.9(C-3″′),131.3(C-4″′),114.9(C-5″′),146.4(C-6″′),146.6(C-7″′),116.9(C-8″′),129.0(C-9″′)。
化合物Z48为6″-O-[(E)-肉桂基]京尼平龙胆二糖苷(6″-O-[(E)-cinnamoyl]-gentio-biosylgenipin):淡黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:5.19(1H,brs,H-1),7.52(1H,brs,H-3),3.03(1H,m,H-5),2.65(1H,m,H-6a),1.94(1H,m,H-6b),5.62(1H,brs,H-7),2.63(1H,m,H-9),3.61(3H,s,11-OCH3),5.02(1H,d,J=7.2Hz,Glc-H-1′),4.57(1H,d,J=7.6Hz,Glc-H-1″),4.33(1H,dd,J=2.0,12.0Hz,Glc-H-6′a),4.13(1H,d,J=2.0,12.0Hz,Glc-H-6′b),4.28(1H,dd,J=2.0,12.0Hz,Glc-H-6″a),4.05(1H,dd,J=2.0,12.0Hz,Glc-H-6″b),6.36(1H,d,J=16.0Hz,H-2″′),7.65(1H,d,J=16.0Hz,H-3″′),7.49(1H,d,J=4.8Hz,7″′),7.46(2H,dd,J=4.6,8.8Hz,H-5″′,9″′),6.76(2H,dd,J=1.2,8.8Hz,H-6″′,8″′)。
9.3栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中60%乙醇洗脱有效部位中黄酮类波谱数据信息:
化合物Z49为5,7-二羟基-3′,4′,5′-三甲氧基黄酮(5,7-dihydroxyl-3′,4′,5′-trimethoxyflavone):黄棕色粉末(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:12.86(1H,s,5-OH),10.89(1H,s,7-OH),7.35(2H,s,H-2′,6′),7.09(1H,s,H-3),6.59(1H,d,J=2.4Hz,H-8),6.23(1H,d,J=2.4Hz,H-6),3.91(6H,s,3′,5′-OCH3),3.75(3H,s,4′-OCH3)。
化合物Z50为5,7,4′-三羟基-6-甲氧基黄酮(5,7,4′-trihydroxyl-6-methoxyflavone):黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:12.94(1H,s,5-OH),10.84(1H,s,7-OH),9.68(1H,brs,4′-OH),8.08(2H,dd,J=2.0,8.4Hz,H-2′,6′),7.60(2H,dd,J=2.0,8.4Hz,H-3′,5′),6.99(1H,s,H-3),6.65(1H,s,H-8),3.76(3H,s,6-OCH3)。
化合物Z51为5,7,4′-三羟基-8-甲氧基黄酮(5,7,4′-trihydroxyl-8-methoxy-flavone):黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:12.51(1H,s,5-OH),8.08(2H,dd,J=2.0,8.4Hz,H-2′,6′),7.63(2H,dd,J=2.0,8.4Hz,H-3′,5′),6.98(1H,s,H-3),6.29(1H,s,H-6),3.85(3H,s,8-OCH3)。
化合物Z52为5,7,3′-三羟基-8,4′,5′-三甲氧基黄酮(5,7,3′-trihydroxyl-8,4′,5′-trimethoxyflavone)。黄色固体(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:12.93(1H,s,5-OH),7.14(1H,s,H-2′),7.13(1H,s,H-6′)6.84(1H,s,H-3),6.45(1H,s,H-6),3.88(3H,s,5′-OCH3),3.74(3H,s,8-OCH3),3.73(3H,s,4′-OCH3)。
化合物Z53为5-羟基-6,7,3′,4′,5′-五甲氧基黄酮(5-hydroxyl-6,7,3′,4′,5′-pentamethoxyflavone):淡黄色结晶(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:12.85(1H,s,5-OH),7.40(2H,s,H-2′,6′),7.17(1H,s,H-3),7.04(1H,s,H-8),3.92(6H,s,3′,5′-OCH3),3.90(3H,s,7-OCH3),3.75(3H,s,4′-OCH3),3.74(3H,s,6-OCH3)。
化合物Z54为5,3′-二羟基-7,4′,5′-三甲氧基黄酮(5,3′-dihydroxyl-7,4′,5′-trimethoxyflavone):淡黄色结晶(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:12.87(1H,s,5-OH),7.21(1H,d,J=2.0Hz,H-2′),7.20(1H,d,J=2.0,H-6′),6.99(1H,s,H-3),6.78(1H,d,J=1.8Hz,H-8),6.40(1H,d,J=1.8Hz,H-6),3.90(3H,s,5′-OCH3),3.89(3H,s,7-OCH3),3.76(3H,s,4′-OCH3)。化合物Z55为5-羟基-7,3′,4′,5′-四甲氧基黄酮(5-hydroxyl-7,3′,4′,5′-tetramethoxyflavone):淡黄色结晶(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:12.86(1H,s,5-OH),7.39(2H,s,H-2′,6′),7.17(1H,s,H-3),6.90(1H,d,J=2.0Hz,H-8),6.41(1H,d,J=2.0Hz,H-6),3.92(6H,s,3′,5′-OCH3),3.90(3H,s,7-OCH3),3.76(3H,s,4′-OCH3)。
化合物Z56为3,5,6,4′-四羟基-3′,5′-二甲氧基黄酮(3,5,6,4′-tetrahydroxy-3′,5′-dimethoxyflavone):黄色粉末(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:9.63(1H,s,3-OH),7.43(1H,d,J=8.0Hz,H-7),7.10(2H,s,H-2′,6′),6.76(1H,d,J=8.0Hz,H-8),3.79(6H,s,3′,5′-OCH3)。
化合物Z57为5,7,3′,4′,5′-五甲氧基黄酮(5,7,3′,4′,5′-pentame-thoxyflavone):淡黄色结晶(甲醇)。1H-NMR(DMSO-d6,400MHz)δ:7.39(2H,s,H-2′,6′),7.17(1H,s,H-3),6.89(1H,s,H-8),6.41(1H,s,H-6),3.92(6H,s,3′,5′-OCH3),3.89(3H,s,7-OCH3),3.88(3H,s,5-OCH3),3.76(3H,s,4′-OCH3)。
9.4栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中60%乙醇洗脱有效部位中二萜类波谱数据信息:
化合物Z58为西红花苷-I(crocin-I):红色粉末,FAB-MS m/z:978[M+H]+。1H-NMR(DMSO-d6,400MHz)δ:7.37(2H,d,J=10.7Hz,H-10,10′),6.88(2H,dd,J=8.0,2.6Hz,H-15,15′),6.83(2H,d,J=14.9Hz,H-12,12′),6.66(2H,dd,J=11.6,11.6Hz,H-11,11′),6.53(2H,brd,J=8.0,H-14,14′),5.42(2H,d,J=7.6Hz,H-1,1″),4.16(2H,d,J=7.8Hz,H-1′,1″′),3.56~3.67(4H,m,H-6,6′,6″,6″′),2.93~3.44(m,糖上其它质子),2.00(6H,s,20,20′-Me),1.97(6H,s,19,19′-Me)。
化合物Z59为西红花苷-II(crocin-II):红色粉末,FAB-MS m/z:816[M+H]+。1H-NMR(DMSO-d6,400MHz)δ:7.37(2H,d,J=11.2Hz,H-10,10′),6.88(2H,dd,J=8.0,2.6Hz,H-15,15′),6.82(2H,dd,J=14.8,2.8Hz,H-12,12′),6.66(2H,dd,J=14.8,11.6Hz,H-11,11′),6.53(2H,brd,J=8.0Hz,H-14,14′),5.42(2H,d,J=7.6Hz,H-1,1″),4.16(1H,d,J=8.0Hz,H-1′),3.56~3.67(3H,m,H-6,6′,6″),2.93~3.48(m,糖上其它质子),2.00(6H,s,20,20′-Me),1.97(6H,s,19,19′-Me)。
实施例10
阿尔茨海默病(AD)的病理特征为大脑皮质和海马出现大量的老年斑(SP)、神经纤维缠结(NFT)、神经元大量丢失和痴呆表现,其中Aβ聚集是中心环节,也是AD发病的关键性因素和关键性治疗靶点,干扰Aβ的产生和阻止其聚集是防治AD的有效途径,开发抑制Aβ纤丝形成和聚集的新药对预防和对症治疗AD具有良好的学术与临床应用价值。本发明利用Aβ25-35致PC12细胞损伤模型,采用MTT法检测了各实验组细胞的活力筛选并确证栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中30%乙醇洗脱有效部位中各单体化合物京尼平苷(Z1)、去乙酰车叶草苷酸甲酯(Z2)、gardaloside(Z3)、6-甲氧基鸡屎藤次苷甲酯(Z4)、6-甲氧基去乙酰车叶草苷酸甲酯(Z5)、栀子苷(Z6)、ajugol(Z7)、genipin-1-β-gentiobiside(Z8)、genipin-1,10-di-O-β-D-glucopyranoside(Z9)、gardoside methyl ester(Z10)、shanzhiside(Z11)、Lamalbidicacid(Z12)、4-carboxyl-7-hydroxy-catapol acid(Z13)、4-carboxyl-catapol acid(Z14)、Mussaenosidic acid(Z15)、geniposidic acid(Z16)、6-hydroxymethy-jasminoside D(Z17)、bornyl-6-O-β-D-xylopyranosyl-β-D-glucopyranoside(Z18)、jasminoside A(Z19)、epijasminoside A(Z20)、jasminoside B(Z21)、jasminoside E(Z22)、jasminoside G(Z23)、jasminoside(Z24)具有治疗阿尔茨海默病(AD)或辅助改善记忆功效的有效成分(波谱数据祥见实施例8中所述化合物),为栀子果实药材治疗阿尔茨海默病(AD)药物或辅助改善记忆功能保健食品的研究奠定理论基础。
1.药物制备
(1)1mg的Aβ25-35加超纯水4.715ml配成200μmol/L,37℃孵育24小时后,置于2-8℃冰箱中贮存,备用。
(2)栀子果实30%乙醇洗脱有效部位中各单体化合物京尼平苷(Z1)、去乙酰车叶草苷酸甲酯(Z2)、gardaloside(Z3)、6-甲氧基鸡屎藤次苷甲酯(Z4)、6-甲氧基去乙酰车叶草苷酸甲酯(Z5)、栀子苷(Z6)、ajugol(Z7)、genipin-1-β-gentiobiside(Z8)、genipin-1,10-di-O-β-D-glucopyranoside(Z9)、gardoside methyl ester(Z10)、shanzhiside(Z11)、Lamalbidicacid(Z12)、4-carboxyl-7-hydroxy-catapol acid(Z13)、4-carboxyl-catapolacid(Z14)、Mussaenosidic acid(Z15)、geniposidic acid(Z16)、6-hydroxymethy-jasminoside D(Z17)、bornyl-6-O-β-D-xylopyranosyl-β-D-glucopyranoside(Z18)、jasminoside A(Z19)、epijasminoside A(Z20)、jasminoside B(Z21)、jasminoside E(Z22)、jasminoside G(Z23)、jasminoside(Z24)的水溶液:将单萜苷类化合物单体Z1~Z24用蒸馏水配成5mmol/L的贮备液,实验时用无血清的DMEM培养液依次稀释成8μmol/L、20μmol/L、100μmol/L的工作液。
2.不同浓度的Aβ25-35对PC12细胞活力的测定,确定造模浓度和作用时间:取培养瓶中处于对数生长期的PC12细胞,经0.25%胰蛋白酶EDTA消化后。用含10%胎牛血清的DMEM高糖培养基将细胞稀释为每毫升悬液含2×105个细胞,接种于96孔培养板,每孔100μl(2×104),放入CO2培养箱,37℃、5%CO2条件下培养,待细胞长满孔底后即可用于实验。实验共分4组,每组8孔,实验时吸弃旧培养液。空白对照组(Control):每孔只加入100μl无血清DMEM低糖培养基;Aβ(25-35)模型组:每孔分别加入用无血清DMEM低糖培养基配制的终浓度为5μM、10μM、20μM的Aβ(25-35)或10μM、20μM、40μM的Aβ(25-35)。37℃培养24h后或48h后,每孔加入终浓度为0.5mg/ml MTT(10μl),继续培养4h后吸去DMEM培养基,每孔加入DMSO100μl,振摇混匀10min,待孔内颗粒完全溶解后,在酶标仪492nm处测定吸光度(A492nm)值。
3.栀子果实30%乙醇洗脱有效部位中各单萜苷类单体化合物Z1~Z24对Aβ25-35所致PC12细胞损伤的保护作用:细胞培养同2.不同浓度的Aβ25-35对PC12细胞活力的测定。实验共分30组,每组10孔,实验时吸弃旧培养液。空白对照组(Control):每孔只加入100μl无血清DMEM低糖培养基;Aβ(25-35)模型组:每孔加入用无血清DMEM低糖培养基配制的终浓度为10μM的Aβ(25-35);各给药组每孔加入终浓度为10μM的Aβ(25-35)及溶液为1.药物制备中的Z1~Z24中各给药组的药物。37℃培养24h后,每孔加入终浓度为0.5mg/ml MTT(10μl),继续培养4h后吸去DMEM培养基,每孔加入DMSO100μl,振摇混匀10min,待孔内颗粒完全溶解后,在酶标仪492nm处测定吸光度(A492nm)值。
4.计算公式
所有数据以表示,组间比较用t检验。细胞存活率的计算公式为:实验组A值/对照组A值×100%。
5实验结果
5.1不同浓度的Aβ25-35对PC12细胞活力的影响和确定造模浓度和作用时间
5μM、10μM、20μM不同浓度的Aβ25-35与PC12细胞共孵24h后,细胞活力显著下降,表现为A492nm值明显下降;且随着Aβ25-35剂量的增加PC12细胞活力下降更加明显。结果见表4。
表4 24h后不同浓度的Aβ25‐35对PC12细胞活力的影响(n=8)
注:**与空白对照组比较p<0.01
10μM、20μM、40μM不同浓度的Aβ25-35与PC12细胞共孵24h后或48h后,细胞活力显著下降,表现为A492nm值明显下降;且随着Aβ25-35剂量和时间的增加PC12细胞活力下降更加明显。结果见表5和6。
表5 24h后不同浓度的Aβ25‐35对PC12细胞活力的影响(n=10)
注:**与空白对照组比较p<0.01
表6 48h后不同浓度的Aβ25‐35对PC12细胞活力的影响(n=10)
注:**与空白对照组比较p<0.01
观察结果:20μM的Aβ25-35处理PC12细胞24小时后,细胞形态发生了很大的改变:模型组PC12细胞可见细胞膜外围的纤毛消失,细胞浆内出现空泡,细胞聚集成团,大多数细胞由多边形变成不规则形状和圆形,细胞的折光性变差。正常对照组的PC12细胞,细胞个体饱满,凸出明显,细胞呈现多边形、菱形或梭形,细胞长势很好,细胞的折光性正常。从而说明Aβ25-35致PC12细胞损伤模型较好的复制了AD的病理变化,是研究AD较好的模型。确定造模浓度和作用时间为:20μM的Aβ25-35处理PC12细胞24小时。
5.2不同浓度的栀子果实30%乙醇洗脱有效部位中各单萜苷类Z1~Z24单体化合物对Aβ25-35所致PC12细胞损伤的保护作用
不同浓度的栀子果实30%乙醇洗脱有效部位中各单体化合物京尼平苷(Z1)、去乙酰车叶草苷酸甲酯(Z2)、gardaloside(Z3)、6-甲氧基鸡屎藤次苷甲酯(Z4)、6-甲氧基去乙酰车叶草苷酸甲酯(Z5)、栀子苷(Z6)、ajugol(Z7)、genipin-1-β-gentiobiside(Z8)、genipin-1,10-di-O-β-D-glucopyranoside(Z9)、gardoside methyl ester(Z10)、shanzhiside(Z11)、Lamalbidicacid(Z12)、4-carboxyl-7-hydroxy-catapol acid(Z13)、4-carboxyl-catapol acid(Z14)、Mussaenosidic acid(Z15)、geniposidic acid(Z16)、6-hydroxymethy-jasminoside D(Z17)、bornyl-6-O-β-D-xylopyranosyl-β-D-glucopyranoside(Z18)、jasminoside A(Z19)、epijasminoside A(Z20)、jasminoside B(Z21)、jasminoside E(Z22)、jasminoside G(Z23)、jasminoside(Z24)对Aβ25-35所致PC12细胞损伤有保护作用,使其细胞活力增强,表现为A492nm值上升。结果见表7、8、9和图1、2、3。不同浓度的单萜苷类单体化合物Z1~Z24对受损的PC12细胞的保护作用如图1、2、3所示。
表7不同浓度的单萜苷类单体化合物Z1~Z8对受损的PC12细胞的保护作用(n=10)
注:*与Aβ模型组比较p<0.05;**与Aβ模型组比较p<0.01
表8不同浓度的单萜苷类单体化合物Z9~Z16对受损的PC12细胞的保护作用(n=10)
注:*与Aβ模型组比较p<0.05;**与Aβ模型组比较p<0.01
表9不同浓度的单萜苷类单体化合物Z17~Z24对受损的PC12细胞的保护作用(n=10)
注:*与Aβ模型组比较p<0.05;**与Aβ模型组比较p<0.01
实施例11
阿尔茨海默病(AD)的病理特征为大脑皮质和海马出现大量的老年斑(SP)、神经纤维缠结(NFT)、神经元大量丢失和痴呆表现,其中Aβ聚集是中心环节,也是AD发病的关键性因素和关键性治疗靶点,干扰Aβ的产生和阻止其聚集是防治AD的有效途径,开发抑制Aβ纤丝形成和聚集的新药对预防和对症治疗AD具有良好的学术与临床应用价值。本发明利用Aβ25-35致PC12细胞损伤模型,采用MTT法检测了各实验组细胞的活力筛选并确证栀子果实治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中60%乙醇洗脱有效部位中各单体化合物单萜苷元类成分jasminoside C aglycone(Z25)、5,5-dihydroxymethyl-1-cyclohexene-furan lactone(Z26)、6-hydroxyl-3-hydroxy-methyl-4,4-dimethyl-1-cyclohexene-1-formaldehyde(Z27)、4-hydroxyl-2-hydroxymethyl-6,6-dimethyl-1-cyclohexene-1-formaldehyde(Z28)、4-hydroxyl-1,5,5-trimethyl-6-epoxyethyl-cyclohex-3-enone(Z29)、iridoids 2a(Z30)、iridoids 2b(Z30)、jasminodiol(Z31)、crocusatin-C(Z32)、7-Epi-crocusatin-C(Z33)、(10S,11S)-Gardendiol(Z34)、(10R,11R)-Gardendiol(Z35)、(5S,9S)-Gardenate A(Z36)、(5R,9R)-Gardenate A(Z36)、5,6-dihydroxymethyl-1,1-dimethylcyclohex-4-enone(Z37)、1-hydroxy-7-hydroxylmethyl-1,4a,5,7a-tetrahydro-pyrancyclopenta-4-carbaldehyde(Z38)或环烯醚萜苯丙素苷类成分5-aldehyde-6′-O-sinapoyljasminoside A(Z39)、6′-O-sinapoyljasminoside B(Z40)、6′-O-trans-sinapoyljasminoside L(Z41)、6′-O-sinapoyl-3-methoxy-4-deketone-jasminoside E(Z42)、6″-O-trans-sinapoylgeni-pingentiobioside(Z43)、6″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin(Z44)、6″-O-[3″′-methoxy-(E)-caffeoyl]-gentiobiosylgenipin(Z45)、4″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin(Z46)、6″-O-[(E)-feruloyl]-gentiobiosylgenipin(Z47)、6″-O-[(E)-cinnamoyl]-gentiobiosylgenipin(Z48)或黄酮类成分5,7-dihydroxyl-3′,4′,5′-trimethoxyflavone(Z49)、5,7,4′-trihydroxyl-6-methoxyflavone(Z50)、5,7,4′-trihydroxyl-8-methoxyflavone(Z51)、5,7,3′-trihydroxyl-8,4′,5′-trimethoxyflavone(Z52)、5-hydroxyl-6,7,3′,4′,5′-pentamethoxyflavone(Z53)、5,3′-dihydroxyl-7,4′,5′-trimethoxyflavone(Z54)、5-hydroxyl-7,3′,4′,5′-tetramethoxyflavone(Z55)、3,5,6,4′-tetrahydroxy-3′,5′-dimethoxyflavone(Z56)、5,7,3′,4′,5′-pentame-thoxyflavone(Z57)或二萜类成分crocin-I(Z58)、crocin-II(Z59)单体化合物具有治疗阿尔茨海默病(AD)或辅助改善记忆功效的有效成分(波谱数据祥见实施例9中所述化合物),为栀子果实药材治疗阿尔茨海默病(AD)药物或辅助改善记忆功能保健食品的研究奠定理论基础。
1.药物制备
(1)1mg的Aβ25-35加超纯水4.715ml配成200μmol/L,37℃孵育24小时后,置于2-8℃冰箱中贮存,备用。
(2)栀子果实60%乙醇洗脱有效部位中各单体化合物单萜类苷元成分jasmino-side C aglycone(Z25)、5,5-dihydroxymethyl-1-cyclohexene-furan lactone(Z26)、6-hydroxyl-3-hydroxy-methyl-4,4-dimethyl-1-cyclohexene-1-formaldehyde(Z27)、4-hydroxyl-2-hydroxymethyl-6,6-dimethyl-1-cyclohexene-1-formaldehyde(Z28)、4-hydroxyl-1,5,5-trimethyl-6-epoxyethyl-cyclohex-3-enone(Z29)、iridoids 2a(Z30)、iridoids 2b(Z30)、jasminodiol(Z31)、crocusatin-C(Z32)、7-Epi-crocusatin-C(Z33)、(10S,11S)-Gardendiol(Z34)、(10R,11R)-Gardendiol(Z35)、(5S,9S)-Gardenate A(Z36)、(5R,9R)-Gardenate A(Z36)、5,6-dihydroxymethyl-1,1-dimethyl-cyclohex-4-enone(Z37)、1-hydroxy-7-hydroxylmethyl-1,4a,5,7a-tetrahydro-pyran-cyclopenta-4-carbaldehyde(Z38)或环烯醚萜苯丙素苷类成分5-aldehyde-6′-O-sinapoyljasminosideA(Z39)、6′-O-sinapoyljasminoside B(Z40)、6′-O-trans-sinapoyljasminoside L(Z41)、6′-O-sinapoyl-3-methoxy-4-deketone-jasminoside E(Z42)、6″-O-trans-sinapoylgenipin gentiobioside(Z43)、6″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin(Z44)、6″-O-[3″′-methoxy-(E)-caffeoyl]-gentiobiosylgenipin(Z45)、4″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin(Z46)、6″-O-[(E)-feruloyl]-gentiobiosylgenipin(Z47)、6″-O-[(E)-cinnamoyl]-gentiobiosylgenipin(Z48)或黄酮类成分5,7-dihydroxyl-3′,4′,5′-trimethoxyflavone(Z49)、5,7,4′-trihydroxyl-6-methoxyflavone(Z50)、5,7,4′-trihydroxyl-8-methoxyflavone(Z51)、5,7,3′-trihydroxyl-8,4′,5′-trimethoxyflavone(Z52)、5-hydroxyl-6,7,3′,4′,5′-pentamethoxyflavone(Z53)、5,3′-dihydroxyl-7,4′,5′-trimethoxyflavone(Z54)、5-hydroxyl-7,3′,4′,5′-tetramethoxyflavone(Z55)、3,5,6,4′-tetrahydroxy-3′,5′-dimethoxyflavone(Z56)、5,7,3′,4′,5′-pentame-thoxyflavone(Z57)或二萜类成分crocin-I(Z58)、crocin-II(Z59)的水溶液:将上述化合物单体Z25~Z59用蒸馏水配成5mmol/L的贮备液,实验时用无血清的DMEM培养液依次稀释成8μmol/L或10μmol/L、20μmol/L或25μmol/L、100μmol/L的工作液。
2.不同浓度的Aβ25-35对PC12细胞活力的测定,确定造模浓度和作用时间见实施例10。
3.栀子果实60%乙醇洗脱有效部位中Z25~Z59单体化合物对Aβ25-35所致PC12细胞损伤的保护作用
细胞培养同实施例10中2.不同浓度的Aβ25-35对PC12细胞活力的测定。实验共分37组,每组10孔,实验时吸弃旧培养液。空白对照组(Control):每孔只加入100μl无血清DMEM低糖培养基;Aβ(25-35)模型组:每孔加入用无血清DMEM低糖培养基配制的终浓度为10μM的Aβ(25-35);各给药组每孔加入终浓度为10μM的Aβ(25-35)及溶液为1.药物制备中的Z25~Z59中各给药组的药物。37℃培养24h后,每孔加入终浓度为0.5mg/ml MTT(10μl),继续培养4h后吸去DMEM培养基,每孔加入DMSO100μl,振摇混匀10min,待孔内颗粒完全溶解后,在酶标仪492nm处测定吸光度(A492nm)值。
4.计算公式
所有数据以表示,组间比较用t检验。细胞存活率的计算公式为:实验组A值/对照组A值×100%。
5实验结果
5.1不同浓度的Aβ25-35对PC12细胞活力的影响和确定造模浓度和作用时间见实施例10。
5.2不同浓度的栀子果实60%乙醇洗脱有效部位中单萜类苷元单体化合物Z25~Z38对Aβ25-35所致PC12细胞损伤的保护作用
不同浓度的栀子果实60%乙醇洗脱有效部位中单萜类苷元成分jasminoside Caglycone(Z25)、5,5-dihydroxymethyl-1-cyclohexene-furan lactone(Z26)、6-hydroxyl-3-hydroxy-methyl-4,4-dimethyl-1-cyclohexene-1-formaldehyde(Z27)、4-hydroxyl-2-hydroxymethyl-6,6-dimethyl-1-cyclohexene-1-formaldehyde(Z28)、4-hydroxyl-1,5,5-trimethyl-6-epoxyethyl-cyclohex-3-enone(Z29)、iridoids 2a(Z30)、iridoids 2b(Z30)、jasminodiol(Z31)、crocusatin-C(Z32)、7-Epi-crocusatin-C(Z33)、(10S,11S)-Gardendiol(Z34)、(10R,11R)-Gardendiol(Z35)、(5S,9S)-Gardenate A(Z36)、(5R,9R)-Gardenate A(Z36)、5,6-dihydroxymethyl-1,1-dimethyl-cyclohex-4-enone(Z37)、1-hydroxy-7-hydroxyl-methyl-1,4a,5,7a-tetrahydro-pyran-cyclopenta-4-carbaldehyde(Z38)单体化合物对Aβ25-35所致PC12细胞损伤有保护作用,使其细胞活力增强,表现为A492nm值上升。结果见表10、11和图4、5。不同浓度的单萜类苷元单体化合物Z25~Z38对受损的PC12细胞的保护作用如图4、5所示。
表10不同浓度的单萜类苷元单体化合物Z25~Z30对受损的PC12细胞的保护作用(n=10)
注:*与Aβ模型组比较p<0.05;**与Aβ模型组比较p<0.01
表11不同浓度的单萜类苷元单体化合物Z31~Z38对受损的PC12细胞的保护作用(n=10)
注:*与Aβ模型组比较p<0.05;**与Aβ模型组比较p<0.01
5.3不同浓度的栀子果实60%乙醇洗脱有效部位中环烯醚萜苯丙素苷类单体化合物Z39~Z48对Aβ25-35所致PC12细胞损伤的保护作用
不同浓度的栀子果实60%乙醇洗脱有效部位中环烯醚萜苯丙素苷类成分5-aldehyde-6′-O-sinapoyljasminoside A(Z39)、6′-O-sinapoyljasminoside B(Z40)、6′-O-trans-sinapoyljasminoside L(Z41)、6′-O-sinapoyl-3-methoxy-4-deketone-jasmi-noside E(Z42)、6″-O-trans-sinapoylgenipin gentiobioside(Z43)、6″-O-[(E)-p-cou-maroyl]-gentiobiosylgenipin(Z44)、6″-O-[3″′-methoxy-(E)-caffeoyl]-gentiobiosyl-genipin(Z45)、4″-O-[(E)-p-coumaroyl]-gentiobiosylgenipin(Z46)、6″-O-[(E)-feruloyl]-gentiobiosylgenipin(Z47)、6″-O-[(E)-cinnamoyl]-gentiobiosylgenipin(Z48)单体化合物对Aβ25-35所致PC12细胞损伤有保护作用,使其细胞活力增强,表现为A492nm值上升。结果见表12和图6。不同浓度的环烯醚萜苯丙素苷类单体化合物Z39~Z48对受损的PC12细胞的保护作用如图6所示。
表12不同浓度的环烯醚萜苯丙素苷类单体化合物Z39~Z48对受损的PC12细胞的保护作用(n=10)
注:*与Aβ模型组比较p<0.05;**与Aβ模型组比较p<0.01
5.4不同浓度的栀子果实60%乙醇洗脱有效部位中黄酮类单体化合物Z49~Z57对Aβ25-35所致PC12细胞损伤的保护作用
不同浓度的栀子果实60%乙醇洗脱有效部位中黄酮类成分5,7-dihydroxyl-3′,4′,5′-trimethoxyflavone(Z49)、5,7,4′-trihydroxyl-6-methoxyflavone(Z50)、5,7,4′-trihydroxyl-8-methoxyflavone(Z51)、5,7,3′-trihydroxyl-8,4′,5′-trimethoxyflavone(Z52)、5-hydroxyl-6,7,3′,4′,5′-pentamethoxyflavone(Z53)、5,3′-dihydroxyl-7,4′,5′-trimethoxyflavone(Z54)、5-hydroxyl-7,3′,4′,5′-tetramethoxyflavone(Z55)、3,5,6,4′-tetrahydroxy-3′,5′-dimethoxyflavone(Z56)、5,7,3′,4′,5′-pentame-thoxyflavone(Z57)单体化合物对Aβ25-35所致PC12细胞损伤有保护作用,使其细胞活力增强,表现为A492nm值上升。结果见表13和图7。不同浓度的黄酮类单体化合物Z49~Z57对受损的PC12细胞的保护作用如图7所示。
表13不同浓度的黄酮类单体化合物Z49~Z57对受损的PC12细胞的保护作用(n=10)
注:*与Aβ模型组比较p<0.05;**与Aβ模型组比较p<0.01
5.5不同浓度的栀子果实60%乙醇洗脱有效部位中二萜类单体化合物Z58~Z59对Aβ25-35所致PC12细胞损伤的保护作用
不同浓度的栀子果实60%乙醇洗脱有效部位中二萜类成分单体化合物crocin-I(Z58)、crocin-II(Z59)对Aβ25-35所致PC12细胞损伤有保护作用,使其细胞活力增强,表现为A492nm值上升。结果见表14和图8。不同浓度的藏红花素二萜类单体化合物Z58~Z59对受损的PC12细胞的保护作用如图8所示。
表14不同浓度的藏红花素二萜类单体化合物Z58~Z59对受损的PC12细胞的保护作用(n=10)
注:*与Aβ模型组比较p<0.05;**与Aβ模型组比较p<0.01 。
Claims (10)
1.栀子果实有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用,其特征在于:该栀子果实药材上述有效部位或有效成分本身是栀子果实总提物上大孔吸附树脂的30%或60%乙醇洗脱部位并具有治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品,及其栀子果实30%或60%乙醇洗脱有效部位分离纯化精制得到萜类或黄酮类单体化合物并具有抑制Aβ纤丝形成和阻止其聚集的生物活性;因拥有共同的环烯醚萜类苷元或单环单萜类苷元或黄酮类苷元或二萜类苷元相同或相似母核结构单元,而具有类似的治疗阿尔茨海默病或辅助改善记忆的功效;其中,栀子药材来源主要为茜草科栀子属植物栀子Gardenia jasminoides Ellis.的干燥成熟果实或茜草科栀子属所有已知栀子属植物的果实。
2.栀子果实30%乙醇洗脱有效部位在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用,其特征在于:该栀子果实药材上述有效部位本身是栀子果实总提物上大孔吸附树脂的30%乙醇洗脱部位经聚酰胺柱色谱纯化得总含量大于90%的总单萜类化合物并具有治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品,因拥有共同的环烯醚萜类苷元、单环单萜类苷元相同或相似母核结构单元,而具有类似的治疗阿尔茨海默病或辅助改善记忆的功效。
3.栀子果实30%乙醇洗脱有效部位中单萜类有效成分在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用,其特征在于:栀子果实30%乙醇洗脱有效部位以单萜苷类成分为主,其总含量大于90%,该栀子果实30%乙醇洗脱有效部位分离纯化精制得到京尼平苷、去乙酰车叶草苷酸甲酯、gardaloside、6-甲氧基鸡屎藤次苷甲酯、6-甲氧基去乙酰车叶草苷酸甲酯、栀子苷、ajugol、genipin-1-β-gentiobiside 、genipin-1,10-di-O -β-D -glucopy-
ranoside、gardoside methyl ester 、shanzhiside 、Lamalbidicacid、4-carboxyl-7-
hydroxy-catapol acid、4-carboxyl-catapol acid、Mussaenosidic acid、geniposidic acid、6-hydroxymethy-jasminoside D 、bornyl-6-O -β-D -xylopyranosyl-β-D -gluco-
pyranoside、jasminoside A、epijasminoside A、jasminoside B、jasminoside E、jasminoside G、jasminoside I中的一种或多种单萜类单体化合物并具有抑制Aβ纤丝形成和阻止其聚集的生物活性,而具有治疗阿尔茨海默病或辅助改善记忆的功效。
4.栀子果实60%乙醇洗脱有效部位在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用,其特征在于:该栀子果实药材上述有效部位本身是栀子果实总提物上大孔吸附树脂的60%乙醇洗脱部位并具有治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品,该栀子果实60%乙醇洗脱有效部位以单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分为主,其上述成分总含量大于90%;因拥有共同的环烯醚萜类苷元、单环单萜类苷元、黄酮类苷元、藏红花素苷元相同或相似母核结构单元,而具有类似的治疗阿尔茨海默病或辅助改善记忆的功效。
5.栀子果实60%乙醇洗脱有效部位中单萜类苷元有效成分在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用,其特征在于:栀子果实60%乙醇洗脱有效部位以单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分为主,其上述成分总含量大于90%,该栀子果实60%乙醇洗脱有效部位分离纯化精制得到jasminoside Caglycone、5,5-dihydroxymethyl -1-cyclohexene-furan lactone、6-hydroxyl-3-hydroxy-methyl-4,4-dimethyl-1-cyc-
lohexene-1-formaldehyde、4-hydroxyl-2-hydroxymethyl-6,6-dimethyl-1-cyclohex-
ene-1-formaldehyde、4-hydroxyl -1,5,5-trimethyl-6-epoxyeth-yl-cyclohex-3-enone、iridoids 2a、iridoids 2b、jasminodiol、crocusatin-C、7-Epi-crocusatin-C、(10S,11S)-Gardendiol、(10R,11R)-Gardendiol、(5S,9S)-Garde-nate A、(5R,9R)-Gardenate A、5,6-dihydroxymethyl-1,1-dimethylcyclohex-4-enone、1-hydroxy-7-hydroxylmethyl-1,4a,5,7a-tetrahydro-pyrancyclopenta-4-carbaldehyde中的一种或多种单萜类苷元单体化合物并具有抑制Aβ纤丝形成和阻止其聚集的生物活性,而具有治疗阿尔茨海默病或辅助改善记忆的功效。
6.栀子果实60%乙醇洗脱有效部位中环烯醚萜苯丙素苷类有效成分在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用,其特征在于:栀子果实60%乙醇洗脱有效部位以单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分为主,其上述成分总含量大于90%,该栀子果实60%乙醇洗脱有效部位分离纯化精制得到5-aldehyde-6′-O-sinapoyljasminoside A、6′-O-sinapoyljasminoside B、6′-O-trans-sinapoyljasminoside L、6′-O-sinapoyl-3-
methoxy-4-deketone-jasminoside E、6′′-O -trans-sinapoylgenipingentiobioside、6′′-O -[(E)-p-coumaroyl]-gentiobiosylgenipin、6′′-O -[3′′′-methoxy-(E)-caffeoyl]-
gentiobiosylgenipin、4′′-O -[(E)-p-coumaroyl]-gentiobiosylgenipin、6′′-O -[(E)-
feruloyl]-gentiobiosylgenipin、6′′-O -[(E)-cinnamoyl]-gentiobiosylgenipin中的一种或多种环烯醚萜苯丙素苷类单体化合物并具有抑制Aβ纤丝形成和阻止其聚集的生物活性,而具有治疗阿尔茨海默病或辅助改善记忆的功效。
7.栀子果实60%乙醇洗脱有效部位中黄酮类苷元有效成分在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用,其特征在于:栀子果实60%乙醇洗脱有效部位以单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分为主,其上述成分总含量大于90%,该栀子果实60%乙醇洗脱有效部位分离纯化精制得到5,7-dihydroxyl-3′,4′,5′-trimethoxyflavone、5,7,4′-trihydroxyl -6- methoxyflavone、5,7,4′-trihydroxyl-8-methoxyflavone、5,7,3′-trihydroxyl-8,4′,5′-trimethoxyflavone、5-hydroxyl-6,7,3′,4′,5′-pentamethoxyflavone、5,3′-dihydroxyl-7,4′,5′-trimethoxyflavone、5-hydroxyl-7,3′,4′,5′-tetramethoxyflavone、3,5,6,4 ′-tetrahydroxy-3′,5′-dimethoxyflavone、5,7,3′,4′,5′-pentame-thoxyflavone中的一种或多种黄酮类苷元单体化合物并具有抑制Aβ纤丝形成和阻止其聚集的生物活性,而具有治疗阿尔茨海默病或辅助改善记忆的功效。
8.栀子果实60%乙醇洗脱有效部位中二萜类有效成分在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用,其特征在于:栀子果实60%乙醇洗脱有效部位以单萜类苷元、环烯醚萜苯丙素苷类、黄酮类、二萜类成分为主,其上述成分总含量大于90%,该栀子果实60%乙醇洗脱有效部位分离纯化精制得到crocin-、crocin-中的一种或多种藏红花素二萜类单体化合物并具有抑制Aβ纤丝形成和阻止其聚集的生物活性,而具有治疗阿尔茨海默病或辅助改善记忆的功效。
9.如权利要求2-8中任意一种或多种栀子果实有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用,其特征在于:所涉及的异质性及多因性阿尔茨海默病或记忆力减退是由衰老、睡眠障碍、心脑血管疾病、神经系统疾病导致的淀粉样蛋白Aβ沉积或Aβ纤丝聚集形成的阿尔茨海默病疾病或记忆障碍或记忆力减退中的任意一种。
10.如权利要求2-9中任意一种或多种栀子果实有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或辅助改善记忆功能保健食品中的应用,其特征在于:所述药物包括口服液、胶囊、片剂、泡腾片、粉针剂、水针剂、注射剂或各种已知剂型制剂或各种可接受剂型制剂,所述药物还包括各种单方和复方制剂,各种已知或各种可接受辅助改善记忆功能类型的保健食品。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610848914.2A CN106619674A (zh) | 2016-09-26 | 2016-09-26 | 栀子有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或保健食品中的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610848914.2A CN106619674A (zh) | 2016-09-26 | 2016-09-26 | 栀子有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或保健食品中的应用 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN106619674A true CN106619674A (zh) | 2017-05-10 |
Family
ID=58854774
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201610848914.2A Pending CN106619674A (zh) | 2016-09-26 | 2016-09-26 | 栀子有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或保健食品中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN106619674A (zh) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107595990A (zh) * | 2017-11-02 | 2018-01-19 | 张忠立 | 一种基于栀子花和代代花的不同浓度乙醇水洗脱部位及其中活性成分组合药物及用途 |
CN107789562A (zh) * | 2017-11-09 | 2018-03-13 | 江西中医药大学 | 一种药食同源的组合物在制备改善睡眠障碍或改善记忆功能减退药物或保健食品中的应用 |
CN110038054A (zh) * | 2019-05-13 | 2019-07-23 | 江西中医药大学 | 栀子花及枝叶果实中有效部位在制备改善睡眠或抗抑郁或改善记忆力减退药物或食品中应用 |
CN115109015A (zh) * | 2022-07-18 | 2022-09-27 | 厦门中药厂有限公司 | C16-Megastigmane类化合物及其制备方法和应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101164571A (zh) * | 2006-10-20 | 2008-04-23 | 中国医学科学院药物研究所 | 栀子有效部位在治疗老年期痴呆中的用途 |
CN101361831A (zh) * | 2008-10-10 | 2009-02-11 | 四川大学华西医院 | 栀子的新用途 |
CN101601677A (zh) * | 2009-07-03 | 2009-12-16 | 江西中医学院 | 一种抗异质性及多因性老年痴呆药物筛选模型的建立及应用 |
-
2016
- 2016-09-26 CN CN201610848914.2A patent/CN106619674A/zh active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101164571A (zh) * | 2006-10-20 | 2008-04-23 | 中国医学科学院药物研究所 | 栀子有效部位在治疗老年期痴呆中的用途 |
CN101361831A (zh) * | 2008-10-10 | 2009-02-11 | 四川大学华西医院 | 栀子的新用途 |
CN101601677A (zh) * | 2009-07-03 | 2009-12-16 | 江西中医学院 | 一种抗异质性及多因性老年痴呆药物筛选模型的建立及应用 |
Non-Patent Citations (2)
Title |
---|
于洋: "栀子抗老年痴呆活性成分研究", 《中国博士学位论文全文数据库医药卫生科技辑》 * |
左月明等: "栀子提取物对异质性及多因性阿尔茨海默病模型大鼠空间学习记忆的影响", 《时珍国医国药》 * |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107595990A (zh) * | 2017-11-02 | 2018-01-19 | 张忠立 | 一种基于栀子花和代代花的不同浓度乙醇水洗脱部位及其中活性成分组合药物及用途 |
CN107789562A (zh) * | 2017-11-09 | 2018-03-13 | 江西中医药大学 | 一种药食同源的组合物在制备改善睡眠障碍或改善记忆功能减退药物或保健食品中的应用 |
CN110038054A (zh) * | 2019-05-13 | 2019-07-23 | 江西中医药大学 | 栀子花及枝叶果实中有效部位在制备改善睡眠或抗抑郁或改善记忆力减退药物或食品中应用 |
CN115109015A (zh) * | 2022-07-18 | 2022-09-27 | 厦门中药厂有限公司 | C16-Megastigmane类化合物及其制备方法和应用 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN105582000B (zh) | 杜仲皮或杜仲叶中萜类物质在制备治疗老年性痴呆或阿尔茨海默病药物中的应用 | |
SG174548A1 (en) | New salvianolic acid compound l, preparation method and use thereof | |
CN106619674A (zh) | 栀子有效部位或有效成分在制备治疗异质性及多因性阿尔茨海默病药物或保健食品中的应用 | |
CN103191174B (zh) | 杜仲化学成分作为血管保护剂的新用途 | |
CN101647850A (zh) | 杜仲化学成分作为植物雌激素的新用途 | |
CN104513290B (zh) | 雷醇内酯衍生物及其应用 | |
CN106008543A (zh) | 一种新的二萜类化合物及其制备方法 | |
KR100733764B1 (ko) | 합환피 추출물 또는 그로부터 분리한 쿠라리디놀을함유하는 고지혈증의 예방 및 치료용 조성물 | |
KR101182199B1 (ko) | 단삼 추출물 또는 크립토탄시논을 유효성분으로 함유하는 뇌졸중의 예방 또는 치료용 조성물 | |
CN101874841A (zh) | 巴戟天属植物总苷提取物及其制备方法和用途 | |
CN109745384A (zh) | 代代花有效部位或有效成分在制备治疗心脑血管系统缺血性疾病药物中的应用及其制备方法 | |
WO2016086842A1 (zh) | 二氢-β-沉香呋喃型倍半萜类化合物、其制法和用途 | |
CN101851162B (zh) | 一种丹酚酸化合物的制备方法和用途 | |
CN106117034A (zh) | 一种高度氧化的倍半萜类化合物及其制备方法和医药用途 | |
CN101683353B (zh) | 黄酮与黄烷衍生物的制备方法及其用途 | |
JP2008506684A (ja) | 「抗ガン性野生チョウセンニンジンの精製エキス調整法及びそのガン融合製剤」 | |
CN108530505A (zh) | 一种黄酮苷类化合物及其制备方法和应用 | |
CN102188459A (zh) | 一种鸦胆子总萜提取物及其制备方法和用途 | |
CN106317002A (zh) | 从大血藤分离出来的天然化合物及其制备方法、应用 | |
Widyasti | Chemical constituents of fan-si abutilon indicum stems | |
KR100242240B1 (ko) | 담배풀로부터 신생혈관 유도현상을 억제하는 세스큐터핀 락톤계 화합물의 제조 방법 및 이를 포함하는 조성물 | |
CN102247354B (zh) | 一种黄烷衍生物的用途 | |
CN102293777B (zh) | 一种黄酮衍生物的用途 | |
CN102302486B (zh) | 一种黄烷衍生物的制药用途 | |
CN106317003A (zh) | 呋喃唑酮的药物组合物及其在生物医药中的应用 |
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 | ||
RJ01 | Rejection of invention patent application after publication | ||
RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170510 |