CN106236792A - 苦丁茶提取物的制备及抗阿尔兹海默症用途 - Google Patents
苦丁茶提取物的制备及抗阿尔兹海默症用途 Download PDFInfo
- Publication number
- CN106236792A CN106236792A CN201610601343.2A CN201610601343A CN106236792A CN 106236792 A CN106236792 A CN 106236792A CN 201610601343 A CN201610601343 A CN 201610601343A CN 106236792 A CN106236792 A CN 106236792A
- Authority
- CN
- China
- Prior art keywords
- active component
- compound
- preparation
- obtains
- extract
- 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.)
- Granted
Links
- 239000000284 extract Substances 0.000 title claims abstract description 24
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 230000007131 anti Alzheimer effect Effects 0.000 title description 2
- 150000001875 compounds Chemical class 0.000 claims abstract description 68
- 239000003814 drug Substances 0.000 claims abstract description 31
- 230000000694 effects Effects 0.000 claims abstract description 28
- 208000024827 Alzheimer disease Diseases 0.000 claims abstract description 14
- 230000004770 neurodegeneration Effects 0.000 claims abstract description 13
- 208000015122 neurodegenerative disease Diseases 0.000 claims abstract description 12
- 238000000746 purification Methods 0.000 claims abstract description 11
- 238000011282 treatment Methods 0.000 claims abstract description 10
- 235000013402 health food Nutrition 0.000 claims abstract description 9
- 230000002265 prevention Effects 0.000 claims abstract description 7
- 238000000605 extraction Methods 0.000 claims abstract description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 78
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 35
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 26
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 21
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 19
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 17
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 14
- 239000000741 silica gel Substances 0.000 claims description 14
- 229910002027 silica gel Inorganic materials 0.000 claims description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 239000000243 solution Substances 0.000 claims description 13
- 238000010829 isocratic elution Methods 0.000 claims description 11
- 239000002904 solvent Substances 0.000 claims description 9
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 claims description 7
- 229940079593 drug Drugs 0.000 claims description 7
- 150000002148 esters Chemical class 0.000 claims description 7
- 238000000926 separation method Methods 0.000 claims description 7
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 230000008859 change Effects 0.000 claims description 5
- 235000013305 food Nutrition 0.000 claims description 4
- 230000000968 intestinal effect Effects 0.000 claims description 3
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 3
- FSLPMRQHCOLESF-UHFFFAOYSA-N alpha-amyrenol Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C)CCC(C)C(C)C5C4=CCC3C21C FSLPMRQHCOLESF-UHFFFAOYSA-N 0.000 claims description 2
- FSLPMRQHCOLESF-SFMCKYFRSA-N alpha-amyrin Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C)CC[C@@H](C)[C@H](C)[C@H]5C4=CC[C@@H]3[C@]21C FSLPMRQHCOLESF-SFMCKYFRSA-N 0.000 claims description 2
- SJMCNAVDHDBMLL-UHFFFAOYSA-N alpha-amyrin Natural products CC1CCC2(C)CCC3(C)C(=CCC4C5(C)CCC(O)CC5CCC34C)C2C1C SJMCNAVDHDBMLL-UHFFFAOYSA-N 0.000 claims description 2
- JFSHUTJDVKUMTJ-QHPUVITPSA-N beta-amyrin Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C JFSHUTJDVKUMTJ-QHPUVITPSA-N 0.000 claims description 2
- QQFMRPIKDLHLKB-UHFFFAOYSA-N beta-amyrin Natural products CC1C2C3=CCC4C5(C)CCC(O)C(C)(C)C5CCC4(C)C3(C)CCC2(C)CCC1(C)C QQFMRPIKDLHLKB-UHFFFAOYSA-N 0.000 claims description 2
- PDNLMONKODEGSE-UHFFFAOYSA-N beta-amyrin acetate Natural products CC(=O)OC1CCC2(C)C(CCC3(C)C4(C)CCC5(C)CCC(C)(C)CC5C4=CCC23C)C1(C)C PDNLMONKODEGSE-UHFFFAOYSA-N 0.000 claims description 2
- WQMLFJWIKARBFW-BKKMTDGVSA-N evomonoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@@H]1C[C@@H](CC[C@H]2[C@]3(CC[C@@H]([C@@]3(C)CC[C@H]32)C=2COC(=O)C=2)O)[C@]3(C)CC1 WQMLFJWIKARBFW-BKKMTDGVSA-N 0.000 claims description 2
- XTLWNMXYCHABQH-UHFFFAOYSA-N germanicol Natural products CC1(C)CCC2(C)CCC3(C)C(CCC4(C)C5(C)CCC(O)C(C)(C)C5CCC34C)C2=C1 XTLWNMXYCHABQH-UHFFFAOYSA-N 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- GGGUGZHBAOMSFJ-GADYQYKKSA-N taraxerol Chemical compound CC([C@@H]1CC2)(C)[C@@H](O)CC[C@]1(C)[C@@H]1[C@]2(C)C2=CC[C@@]3(C)CCC(C)(C)C[C@H]3[C@]2(C)CC1 GGGUGZHBAOMSFJ-GADYQYKKSA-N 0.000 claims description 2
- HYTFUKZLSYXRDX-UHFFFAOYSA-N taraxerol Natural products CC1CCCC2C3(C)CCC4(C)C5CC(C)(C)CCC5(C)C=CC4(C)C3C(O)CC12C HYTFUKZLSYXRDX-UHFFFAOYSA-N 0.000 claims description 2
- XCHZVKNZDNVOLO-UHFFFAOYSA-N 19alpha-Hydroxy-ursolic-acid Natural products CC1CCC2(CCC3(C)C(=CCC4C3(C)CCC5C(C)(C)C(O)CCC45CO)C2C1C)C(=O)O XCHZVKNZDNVOLO-UHFFFAOYSA-N 0.000 claims 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims 1
- 244000046052 Phaseolus vulgaris Species 0.000 claims 1
- JYEJQUHSBMDABO-SYKQLJDKSA-N Pomolic acid Natural products C[C@@H]1CC[C@@]2(CC[C@H]3[C@@H]4CC[C@H]5C(C)(C)[C@@H](O)CC[C@]5(C)[C@H]4CC=C3[C@@H]2[C@]1(C)O)C(=O)O JYEJQUHSBMDABO-SYKQLJDKSA-N 0.000 claims 1
- 238000010298 pulverizing process Methods 0.000 claims 1
- 108010025020 Nerve Growth Factor Proteins 0.000 abstract description 24
- 102000015336 Nerve Growth Factor Human genes 0.000 abstract description 22
- 229940053128 nerve growth factor Drugs 0.000 abstract description 8
- 239000000463 material Substances 0.000 abstract description 7
- 239000002547 new drug Substances 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 22
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 19
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 16
- 150000002966 pentacyclic triterpenoids Chemical class 0.000 description 12
- 210000005036 nerve Anatomy 0.000 description 9
- 238000011160 research Methods 0.000 description 9
- 238000005160 1H NMR spectroscopy Methods 0.000 description 7
- 201000010099 disease Diseases 0.000 description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 239000007787 solid Substances 0.000 description 7
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 6
- 210000002569 neuron Anatomy 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 5
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 5
- 241001597008 Nomeidae Species 0.000 description 5
- 229920002472 Starch Polymers 0.000 description 5
- 244000269722 Thea sinensis Species 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- 239000002552 dosage form Substances 0.000 description 5
- 239000003112 inhibitor Substances 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 239000008107 starch Substances 0.000 description 5
- 235000019698 starch Nutrition 0.000 description 5
- 102100033639 Acetylcholinesterase Human genes 0.000 description 4
- 108010022752 Acetylcholinesterase Proteins 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- ADEBPBSSDYVVLD-UHFFFAOYSA-N donepezil Chemical compound O=C1C=2C=C(OC)C(OC)=CC=2CC1CC(CC1)CCN1CC1=CC=CC=C1 ADEBPBSSDYVVLD-UHFFFAOYSA-N 0.000 description 4
- 239000003937 drug carrier Substances 0.000 description 4
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 4
- 230000014759 maintenance of location Effects 0.000 description 4
- 238000001819 mass spectrum Methods 0.000 description 4
- 239000013641 positive control Substances 0.000 description 4
- 108090000765 processed proteins & peptides Proteins 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- 206010012289 Dementia Diseases 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- ZRJBHWIHUMBLCN-SEQYCRGISA-N Huperzine A Natural products N1C(=O)C=CC2=C1C[C@H]1/C(=C/C)[C@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-SEQYCRGISA-N 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- ZRJBHWIHUMBLCN-UHFFFAOYSA-N Shuangyiping Natural products N1C(=O)C=CC2=C1CC1C(=CC)C2(N)CC(C)=C1 ZRJBHWIHUMBLCN-UHFFFAOYSA-N 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 230000008499 blood brain barrier function Effects 0.000 description 3
- 210000001218 blood-brain barrier Anatomy 0.000 description 3
- 210000004556 brain Anatomy 0.000 description 3
- 230000004069 differentiation Effects 0.000 description 3
- 239000003623 enhancer Substances 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- ZRJBHWIHUMBLCN-YQEJDHNASA-N huperzine A Chemical compound N1C(=O)C=CC2=C1C[C@H]1\C(=C/C)[C@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-YQEJDHNASA-N 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 150000002772 monosaccharides Chemical class 0.000 description 3
- 239000008194 pharmaceutical composition Substances 0.000 description 3
- 239000008363 phosphate buffer Substances 0.000 description 3
- 229920001223 polyethylene glycol Polymers 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 3
- ZRJBHWIHUMBLCN-BMIGLBTASA-N rac-huperzine A Natural products N1C(=O)C=CC2=C1C[C@@H]1C(=CC)[C@@]2(N)CC(C)=C1 ZRJBHWIHUMBLCN-BMIGLBTASA-N 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 2
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 102000007072 Nerve Growth Factors Human genes 0.000 description 2
- RVGRUAULSDPKGF-UHFFFAOYSA-N Poloxamer Chemical compound C1CO1.CC1CO1 RVGRUAULSDPKGF-UHFFFAOYSA-N 0.000 description 2
- 244000173166 Pyrus ussuriensis Species 0.000 description 2
- XSVMFMHYUFZWBK-NSHDSACASA-N Rivastigmine Chemical compound CCN(C)C(=O)OC1=CC=CC([C@H](C)N(C)C)=C1 XSVMFMHYUFZWBK-NSHDSACASA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 206010052428 Wound Diseases 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 229940022698 acetylcholinesterase Drugs 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- PNNNRSAQSRJVSB-BXKVDMCESA-N aldehydo-L-rhamnose Chemical compound C[C@H](O)[C@H](O)[C@@H](O)[C@@H](O)C=O PNNNRSAQSRJVSB-BXKVDMCESA-N 0.000 description 2
- SHZGCJCMOBCMKK-PQMKYFCFSA-N alpha-D-rhamnose Chemical compound C[C@H]1O[C@H](O)[C@@H](O)[C@@H](O)[C@@H]1O SHZGCJCMOBCMKK-PQMKYFCFSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 230000036772 blood pressure Effects 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 230000001713 cholinergic effect Effects 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 230000008021 deposition Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 229960003530 donepezil Drugs 0.000 description 2
- 238000001425 electrospray ionisation time-of-flight mass spectrometry Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 150000004665 fatty acids Chemical class 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000013355 food flavoring agent Nutrition 0.000 description 2
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 2
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 235000010445 lecithin Nutrition 0.000 description 2
- 239000000787 lecithin Substances 0.000 description 2
- 229940067606 lecithin Drugs 0.000 description 2
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 2
- 235000019359 magnesium stearate Nutrition 0.000 description 2
- 150000004702 methyl esters Chemical class 0.000 description 2
- 229940016286 microcrystalline cellulose Drugs 0.000 description 2
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 2
- 239000008108 microcrystalline cellulose Substances 0.000 description 2
- 230000001537 neural effect Effects 0.000 description 2
- 230000000324 neuroprotective effect Effects 0.000 description 2
- 239000003900 neurotrophic factor Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 208000028591 pheochromocytoma Diseases 0.000 description 2
- 229960000502 poloxamer Drugs 0.000 description 2
- 229920001983 poloxamer Polymers 0.000 description 2
- JUJWROOIHBZHMG-RALIUCGRSA-N pyridine-d5 Chemical compound [2H]C1=NC([2H])=C([2H])C([2H])=C1[2H] JUJWROOIHBZHMG-RALIUCGRSA-N 0.000 description 2
- 229960004136 rivastigmine Drugs 0.000 description 2
- 239000002002 slurry Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 229960001685 tacrine Drugs 0.000 description 2
- YLJREFDVOIBQDA-UHFFFAOYSA-N tacrine Chemical compound C1=CC=C2C(N)=C(CCCC3)C3=NC2=C1 YLJREFDVOIBQDA-UHFFFAOYSA-N 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 229930182493 triterpene saponin Natural products 0.000 description 2
- RXPRRQLKFXBCSJ-GIVPXCGWSA-N vincamine Chemical compound C1=CC=C2C(CCN3CCC4)=C5[C@@H]3[C@]4(CC)C[C@](O)(C(=O)OC)N5C2=C1 RXPRRQLKFXBCSJ-GIVPXCGWSA-N 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- UIAGMCDKSXEBJQ-IBGZPJMESA-N 3-o-(2-methoxyethyl) 5-o-propan-2-yl (4s)-2,6-dimethyl-4-(3-nitrophenyl)-1,4-dihydropyridine-3,5-dicarboxylate Chemical compound COCCOC(=O)C1=C(C)NC(C)=C(C(=O)OC(C)C)[C@H]1C1=CC=CC([N+]([O-])=O)=C1 UIAGMCDKSXEBJQ-IBGZPJMESA-N 0.000 description 1
- 206010000117 Abnormal behaviour Diseases 0.000 description 1
- 208000017194 Affective disease Diseases 0.000 description 1
- 206010001497 Agitation Diseases 0.000 description 1
- 102000002659 Amyloid Precursor Protein Secretases Human genes 0.000 description 1
- 108010043324 Amyloid Precursor Protein Secretases Proteins 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- XUJNEKJLAYXESH-UWTATZPHSA-N D-Cysteine Chemical compound SC[C@@H](N)C(O)=O XUJNEKJLAYXESH-UWTATZPHSA-N 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 206010064571 Gene mutation Diseases 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
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- WTDHULULXKLSOZ-UHFFFAOYSA-N Hydroxylamine hydrochloride Chemical compound Cl.ON WTDHULULXKLSOZ-UHFFFAOYSA-N 0.000 description 1
- 241001545416 Ilex kaushue Species 0.000 description 1
- 244000078118 Ilex latifolia Species 0.000 description 1
- SRBFZHDQGSBBOR-HWQSCIPKSA-N L-arabinopyranose Chemical compound O[C@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-HWQSCIPKSA-N 0.000 description 1
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- 208000019022 Mood disease Diseases 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 102000003923 Protein Kinase C Human genes 0.000 description 1
- 108090000315 Protein Kinase C Proteins 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 description 1
- 150000004075 acetic anhydrides Chemical class 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000001919 adrenal effect Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- GZCGUPFRVQAUEE-SLPGGIOYSA-N aldehydo-D-glucose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O GZCGUPFRVQAUEE-SLPGGIOYSA-N 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000003466 anti-cipated effect Effects 0.000 description 1
- 230000001640 apoptogenic effect Effects 0.000 description 1
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 210000001367 artery Anatomy 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 230000003925 brain function Effects 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 229910052804 chromium Inorganic materials 0.000 description 1
- 239000011651 chromium Substances 0.000 description 1
- DERZBLKQOCDDDZ-JLHYYAGUSA-N cinnarizine Chemical compound C1CN(C(C=2C=CC=CC=2)C=2C=CC=CC=2)CCN1C\C=C\C1=CC=CC=C1 DERZBLKQOCDDDZ-JLHYYAGUSA-N 0.000 description 1
- 229960000876 cinnarizine Drugs 0.000 description 1
- 210000001728 clone cell Anatomy 0.000 description 1
- 229940125904 compound 1 Drugs 0.000 description 1
- 229940125782 compound 2 Drugs 0.000 description 1
- 229940126214 compound 3 Drugs 0.000 description 1
- 229940125898 compound 5 Drugs 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 230000006837 decompression Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 230000000994 depressogenic effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 239000007919 dispersible tablet Substances 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- RXPRRQLKFXBCSJ-UHFFFAOYSA-N dl-Vincamin Natural products C1=CC=C2C(CCN3CCC4)=C5C3C4(CC)CC(O)(C(=O)OC)N5C2=C1 RXPRRQLKFXBCSJ-UHFFFAOYSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 210000000232 gallbladder Anatomy 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 230000003862 health status Effects 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 description 1
- 230000006951 hyperphosphorylation Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000003978 infusion fluid Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002608 insulinlike Effects 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 229940040145 liniment Drugs 0.000 description 1
- 239000000865 liniment Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- MQYXUWHLBZFQQO-QGTGJCAVSA-N lupeol Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C)CC[C@@H](C(=C)C)[C@@H]5[C@H]4CC[C@@H]3[C@]21C MQYXUWHLBZFQQO-QGTGJCAVSA-N 0.000 description 1
- PKGKOZOYXQMJNG-UHFFFAOYSA-N lupeol Natural products CC(=C)C1CC2C(C)(CCC3C4(C)CCC5C(C)(C)C(O)CCC5(C)C4CCC23C)C1 PKGKOZOYXQMJNG-UHFFFAOYSA-N 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000002398 materia medica Substances 0.000 description 1
- BUGYDGFZZOZRHP-UHFFFAOYSA-N memantine Chemical compound C1C(C2)CC3(C)CC1(C)CC2(N)C3 BUGYDGFZZOZRHP-UHFFFAOYSA-N 0.000 description 1
- 229960004640 memantine Drugs 0.000 description 1
- 230000029052 metamorphosis Effects 0.000 description 1
- 239000000401 methanolic extract Substances 0.000 description 1
- 210000000274 microglia Anatomy 0.000 description 1
- 150000002771 monosaccharide derivatives Chemical class 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000003928 nasal cavity Anatomy 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 230000007472 neurodevelopment Effects 0.000 description 1
- 229960000715 nimodipine Drugs 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- JUJWROOIHBZHMG-UHFFFAOYSA-O pyridinium Chemical compound C1=CC=[NH+]C=C1 JUJWROOIHBZHMG-UHFFFAOYSA-O 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000011506 response to oxidative stress Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000008299 semisolid dosage form Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 210000000225 synapse Anatomy 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 102000013498 tau Proteins Human genes 0.000 description 1
- 108010026424 tau Proteins Proteins 0.000 description 1
- 230000035922 thirst Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 229960002726 vincamine Drugs 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
Classifications
-
- 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)
-
- 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
- A61K2236/50—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)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
本发明提供一种苦丁茶提取物的制备方法,通过将中药苦丁茶粉碎和浸提,分离和纯化,获得苦丁茶提取物为活性组分(A),(b)‑(f),(B)‑(E),和活性化合物(1)‑(5)。苦丁茶活性组分(A),(b)‑(f),(B)‑(E),活性化合物(1)‑(5)具有显著的拟神经生长因子的活性,可以制备预防阿尔兹海默症等神经退行性疾病的药物和保健食品。本发明中的中药苦丁茶来源广泛,取材便利,廉价易得,所以将苦丁茶开发为预防和治疗阿尔兹海默症等神经退行性疾病的新药和保健食品具有重要意义。
Description
技术领域
本发明属医药技术领域,具体涉及一种苦丁茶提取物及其活性组分和活性化合物的制备方法,以及其在预防及治疗阿尔兹海默症等神经退行性疾病中的食品和药品中的应用。
背景技术
老年人口的快速增长,使得人们更加关心该群体的健康状况。据各国卫生组织统计,仅在欧美各国,有20%的人在一生之中会受到脑功能障碍类疾病的困扰,每年总支出高达2万亿美元,超过心血管疾病、癌症、糖尿病三者的总和,并且预计随着人类寿命的增加,这一数字还会继续增加。尤其是阿尔兹海默症(Alzeimer’s disease,AD),已经成为个人医疗支出增长的主要因素,为社会带来很大负担。在国际阿尔茨海默症协会年报告中指出:2015年,全世界约有4680万人患有痴呆症,预计2030年将达到7470万人,2050年更将突破1亿3150万人,其中约一半为AD患者。58%的痴呆症患者生活在现今的中低收入国家,在2030年这一数字将增加到63%,2050年则达到68%。2015年全球痴呆症照护成本总计为8180亿美元。短短3年,此金额将增加至1兆美元,2030年更会高达2兆美元。
我国阿尔兹海默症患者估计超过500万,约占世界总病例数的1/4;而且,随着我国人口老龄化进程的加快,这个数字将更为庞大,给社会稳定与发展带来重大的影响。据统计,中国阿尔兹海默症的发病率65岁以上是6.6%,年龄每增加5岁,患病率增长一倍,85岁以上则高达40%。再过20年,如今的中年人都将步入老年人的行列,阿尔兹海默症患者数量将急剧增加。因此,研究和开发有效的预防治疗阿尔兹海默症等神经退行性疾病的药物已成为全世界迫切需要解决的医学问题。
长期看来,AD是一种致死性的神经退行性疾病,发病后平均生存时间约为10年。AD是由神经退行性病变、脑血管病变、感染、外伤、肿瘤、营养代谢障碍等多种原因引起的一组症候群,是病人在意识清醒的状态下出现的持久的全面的智能减退,表现为记忆力、计算力、判断力、注意力、抽象思维能力、语言功能减退,情感障碍,日常生活能力和工作能力进行性减退直至丧失,并有各种神经精神症状和行为障碍。病理特征为淀粉样蛋白斑块和神经纤维缠结,在斑块附近存在大量的激活态小胶质细胞,同时脑内大量神经突触和神经元丢失。根据这些特征,目前主要学术假说有以下几种:1.β淀粉样多肽(Aβ)毒性及沉积;2.Tau蛋白过度磷酸化假说3.胆碱能缺失学说;4.神经退行性病变(Neurodegeneration);5.其它多种因素,如基因突变学说、氧化应激学说。
现今用于治疗AD的药物很多,主要有胆碱能药,其中乙酰胆碱酯酶(Acetylcholinesterase,AChE)抑制剂上市的药物有他克林(tacrine)、酒石酸卡巴拉汀(rivastigmine)、石杉碱甲(huperzine A)、多奈哌齐(donepezil)等;抗Aβ蛋白沉积的药物,主要有胰岛素样激素、结晶抑制剂和分泌酶抑制剂;神经兴奋性毒性抑制剂美金刚;脑代谢调节剂,如长春胺、尼莫地平、脑益嗪;影响自由基代谢的药物,如维生素C结合维生素E等。但AD是神经退行性疾病,病程中所造成的神经细胞的死亡是不可逆的,而这些治疗药物只能延缓病理进程,且随病情发展药物疗效逐渐降低,有副作用,因此,近年来寻找新的针对AD病因的治疗药物和方法,成为当今研究的热点和难点。
研究表明,神经营养因子对神经发育和成年神经系统的疾病过程都有重要的影响。在神经退行性病变动物模型中,发现神经生长因子(Nerve Growth Factor,NGF)能阻止或减少神经元的退变。NGF是人类发现的第一个神经营养因子,也是最重要的神经营养因子;是一种对神经细胞的生长、发育、分化和功能保持等方面有重要调控作用的生物活性多肽;对神经萎缩、神经变性、外伤修复等神经疾病的治疗有显著作用。研究发现,NGF一定程度可阻止AD进展,其促进神经生长和神经保护作用是长期的研究热点。然而,它是一个由100多个氨基酸组成的多肽;由于分子量大和极性强等原因,不能通过血脑屏障(BloodBrain Barrier),并且难以大规模制备等诸多因素,局限了它的实际临床应用。目前,NGF除脑内手术直接投药之外,还没有找到更好的治疗方法。因此,寻找具有拟NGF活性(NGFmimics)或能增强其活性(NGF enhancer)并且能通过血脑屏障的小分子化合物就自然成为了研究方向。由于PC12细胞(Pheochromocytoma cells,从大鼠肾上腺嗜铬细胞克隆得到)在NGF的作用下会停止分裂,长出突起,转化成神经元样细胞。因此,是一个很好的在细胞分子水平研究NGF的功能的细胞模型。目前,已经有NGF mimics在III期临床试验阶段。
中草药是中药学的物质基础,是天然活性有机化合物的主要来源。我国有着几千年的中医文明,且地大物博,有很多宝贵的道地药材,因为中药材资源的利用较方便,所以对其新的成分和新的活性的研究也就十分重要。研究表明,中药对阿尔兹海默症有显著疗效,其中最著名的是从蛇足石杉中提取的生物碱-石杉碱甲(huperzine A),它是中国科学院药物研究所开发的具有自主知识产权的创新药物,它不仅是一种高效高选择性的乙酰胆碱酯酶(AChE)抑制药,而且还能减少谷氨酸诱发的神经细胞死亡,保护神经细胞对抗β淀粉样多肽(Aβ)产生的氧化应激反应,具有明显的对抗过氧化氢、蛋白激酶C抑制剂等诱导的神经细胞凋亡的作用,是目前全球治疗阿尔兹海默症的最佳药物之一。
苦丁茶为冬青科冬青属苦丁茶Ilex kudingcha C.J.Tseng的叶,主要分布在西南地区(四川、重庆、贵州、湖南、湖北)及华南地区(江西、广东、福建、海南)等地,是纯天然保健饮料。具有清热消暑、明目益智、生津止渴、利尿强心、润喉止咳、降压减肥、抑癌防癌、抗衰老、活血脉等多种功效,素有“保健茶”、“美容茶”、“减肥茶”、“降压茶”、“益寿茶”等美称。
发明内容
本发明的目的是提供一种苦丁茶提取物的制备方法,通过以下步骤实现:
(1)粉碎和浸提:
100g中药苦丁茶粉碎后,用1L甲醇室温下震荡浸提3次,每次24小时,所得滤液经浓缩干燥后,用水和乙酸乙酯交替萃取分离多次,得到酯层活性组分(A)7g;
(2)分离和纯化:
将活性组分(A)先经一次硅胶开口柱分离(200-300目,以下溶剂系统均按体积比),经正己烷:乙酸乙酯(100:0,70:30,50:50,0:100)洗脱,最后用甲醇冲洗,合并正己烷:乙酸乙酯70:30的洗脱流分,得到活性组分(b)2g;合并甲醇的洗脱流分,得到活性组分(B)2.2g。
活性组分(b)2g经硅胶开口柱分离,正己烷:氯仿3:7洗脱,得1g活性组分(c),继续经硅胶开口柱分离,合并正己烷:氯仿5:5洗脱流分得活性组分(d)0.45g,活性组分(d)经正己烷:氯仿7:3洗脱得活性组分(e)76mg;活性组分(e)用HPLC纯化(色谱条件为:色谱柱流速3ml/min,检测波长210nm,流动相为99%乙腈水溶液等度洗脱)得到化合物(1)10.3mg(tR=38min)、化合物(2)5.3mg(tR=57min)、活性组分(f)12.5mg;活性组分(f)继续用HPLC纯化(色谱条件为:色谱柱流速3ml/min,检测波长210nm,流动相为97%甲醇水溶液等度洗脱)得到化合物(3)3.7mg(tR=37min)、化合物(4)4.5mg(tR=35min)。经核磁共振谱、质谱确定化合物(1)-(4)为五环三萜类化合物。
活性组分(B)进一步用ODS开口柱分离(溶剂系统按体积比为甲醇:水=5:5,7:3,8:2,9:1,10:0),合并甲醇:水为8:2至9:1的洗脱流分,得到活性组分(C)350mg;活性组分(C)再次用硅胶开口柱分离(溶剂系统按体积比为氯仿:乙酸乙酯=95:5,90:10,80:20,70:30,60:40,50:50,40:60,30:70,20:80,90:10,100:0),合并甲醇:水为10:90至0:100的洗脱流分,得到活性组分(D)67mg;活性组分(D)用HPLC纯化(色谱条件为:色谱柱 流速3ml/min,检测波长210nm,流动相为45%乙腈水溶液等度洗脱30分钟),得到活性组分(E)23mg,继续用HPLC纯化(色谱条件为:色谱柱流速3ml/min,检测波长210nm,流动相为63%甲醇水溶液等度洗脱50分钟)得到化合物(5)12.3mg,经核磁共振谱、质谱确定为新五环三萜皂苷类化合物。
本发明制备的活性组分(E),其中五环三萜类化合物(5)的总含量>50%。同时确定了化合物(1)为羽扇豆醇,化合物(2)为α-香树脂醇,化合物(3)为β-香树脂醇,化合物(4)为蒲公英赛醇,化合物(5)为3β-O-D-葡萄糖(1→3)-[α-L-鼠李糖(1→3)]-α-L-阿拉伯糖-坡模酸。(3-O-β-D–glucopyranosyl-(1→3)-[α-L-rhamnopyranosyl-(1→2)]-α-L-arabinopyranosyl)-pomolic acid)。
本发明的另一个目的是提供一种苦丁茶提取物在制备预防或治疗阿尔兹海默症等神经退行性疾病的药物和保健食品中的应用。所述的苦丁茶提取物为活性组分(A),(b)-(f),(B)-(E),和活性化合物(1)-(5)。
苦丁茶活性组分(A),(b)-(f),(B)-(E),活性化合物(1)-(5)具有显著的拟神经生长因子的活性,因此可将其作为有效成分,添加药学或食品可接受的载体或赋形剂,制备预防、治疗阿尔兹海默症等神经退行性疾病的保健食品和药物。所述的保健食品可接受的载体是指食品领域常规的载体,例如淀粉、蔗糖、微晶纤维素等填充剂,淀粉浆、羟丙纤维素、明胶、聚乙二醇等粘合剂,硬脂酸镁、微粉硅胶、聚乙二醇类等湿润剂,聚山梨脂、卵磷脂等吸收促进剂,伯洛沙姆、脂肪酸山梨坦、聚山梨脂等表面活性剂,另外还可以添加香味剂、甜味剂等其它辅剂。活性化合物(1)-(5)结构式如下所示:
所述的药学可接受的载体可参照现有技术。将所述的活性成分或化合物制成药品后,可以以单位剂量形式给用,给药途径可以为肠道或非肠道给药。
所述药物的剂型可以是固体制剂、液体制剂或半固体剂型,上述各种剂型可采用现有生产方法进行制备。
本发明进一步还提供一种预防阿尔兹海默症等神经退行性疾病的药物组合物,该药物组合物含有生理有效量苦丁茶活性组分(A),(b)-(f),(B)-(E),活性化合物(1)-(5)和药学上可接受的载体或稀释剂。
这里所述的药学上可接受的载体是指药学领域常规的药物载体,例如稀释剂、赋形剂如是等,填充剂如淀粉、蔗糖、微晶纤维素等;粘合剂如淀粉浆、羟丙纤维素、明胶、聚乙二醇等;湿润剂如硬脂酸镁、微粉硅胶、聚乙二醇类等;吸收促进剂聚山梨脂、卵磷脂等,表面活性剂伯洛沙姆、脂肪酸山梨坦、聚山梨脂等等,另外还可以在组合物中加入其它辅剂如香味剂、甜味剂等。
本发明所述的苦丁茶活性组分(A),(b)-(f),(B)-(E),活性化合物(1)-(5)可以以单位剂量形式给药,给药途径可为肠道和非肠道,包括口服、肌肉、皮下和鼻腔。
本发明所述的化合物给药途径可为静脉给药。注射包括静脉注射、肌肉注射、皮下注射和穴位注射。
本发明的药物组合物的各种剂型可以按照药学领域的常规生产方法制备,例如使活性成分与一种或多种载体或赋形剂混合,然后将其制成所需的剂型。
给药剂型可以是固体制剂、胶囊剂或液体制剂,包括片剂、胶囊剂、分散片、口服液、大输液、小针、冻干粉针、软膏、搽剂或栓剂。
用本发明制备的苦丁茶活性组分(A),(b)-(f),(B)-(E),活性化合物(1)-(5),具有显著的拟神经生长因子的活性,可以制备预防阿尔兹海默症等神经退行性疾病的药物和保健食品。本发明中的中药材苦丁茶廉价易得,且至今无其具有拟神经生长因子活性的相关报道。因此,本发明对该中药新的药理活性研究及开发具有重要意义。
本发明采用PC 12细胞作为有效的活性鉴定系统,发现苦丁茶甲醇提取物分离得到的活性组分及活性化合物,能诱导PC 12细胞神经突起伸长,具有显著的拟神经生长因子活性。此外,苦丁茶来源广泛,取材便利等优势。所以将苦丁茶开发为预防和治疗阿尔兹海默症等神经退行性疾病的新药和保健食品具有重要意义。
附图说明
图1是加入苦丁茶活性组分(A),(b)-(f),活性化合物(1)-(4),经48小时后PC 12细胞的神经突起分化率。
图2是加入苦丁茶活性组分(A),(b)-(f),活性化合物(1)-(4),经48小时后PC 12细胞显微镜图片。
图3是加入苦丁茶活性组分(A),(B)-(E),活性化合物(5),经48小时后PC 12细胞的神经突起分化率。
图4是加入苦丁茶活性组分(A),(B)-(E),活性化合物(5),经48小时后PC 12细胞显微镜图片。
具体实施方式
本发明结合附图和具体实施例作进一步的说明。
实施例1苦丁茶活性组分和活性化合物的制备
(1)粉碎和浸提:
100g中药苦丁茶粉碎后,用1L甲醇室温下震荡浸提3次,每次24小时,所得滤液经浓缩干燥后,用水和乙酸乙酯交替萃取分离多次,得到酯层活性组分(A)7g;
(2)分离和纯化:
将活性组分(A)先经一次硅胶开口柱分离(200-300目,以下溶剂系统均按体积比),经正己烷:乙酸乙酯(100:0,70:30,50:50,0:100)洗脱,最后用甲醇冲洗,合并正己烷:乙酸乙酯70:30的洗脱流分,得到活性组分(b)2g;合并甲醇的洗脱流分,得到活性组分(B)2.2g。
活性组分(b)2g经硅胶开口柱分离,正己烷:氯仿3:7洗脱,得1g活性组分(c),继续经硅胶开口柱分离,合并正己烷:氯仿5:5洗脱流分得活性组分(d)0.45g,活性组分(d)经正己烷:氯仿7:3洗脱得活性组分(e)76mg;活性组分(e)用HPLC纯化(色谱条件为:色谱柱流速3ml/min,检测波长210nm,流动相为99%乙腈水溶液等度洗脱)得到化合物(1)10.3mg(tR=38min),化合物(2)5.3mg(tR=57min),活性组分(f)12.5mg;活性组分(f)继续用HPLC纯化(色谱条件为:色谱柱流速3ml/min,检测波长210nm,流动相为97%甲醇水溶液等度洗脱)得到化合物(3)3.7mg(tR=37min),化合物(4)4.5mg(tR=35min)。经核磁共振谱、质谱确定为五环三萜类化合物。
活性组分(B)进一步用ODS开口柱分离(溶剂系统按体积比为甲醇:水=5:5,7:3,8:2,9:1,10:0),合并甲醇:水为8:2至9:1的洗脱流分,得到活性组分(C)350mg;活性组分(C)再次用硅胶开口柱分离(溶剂系统按体积比为氯仿:乙酸乙酯=95:5,90:10,80:20,70:30,60:40,50:50,40:60,30:70,20:80,90:10,100:0),合并甲醇:水为10:90至0:100的洗脱流分,得到活性组分(D)67mg;活性组分(D)用HPLC纯化(色谱条件为:色谱柱 流速3ml/min,检测波长210nm,流动相为45%乙腈水溶液等度洗脱30分钟),得到活性组分(E)23mg,继续用HPLC纯化(色谱条件为:色谱柱流速3ml/min,检测波长210nm,流动相为63%甲醇水溶液等度洗脱50分钟)得到化合物(5)12.3mg,经核磁共振谱、质谱确定为新五环三萜皂苷类化合物。
实施例2
对实施例1所得化合物理化特征及化学结构的定性鉴定:
化合物(1)-(4)的结构经LC-MS、1H NMR、13C NMR和文献数据比对后确定。
化合物1:白色固体,C30H50O(HRESI-TOF-MS m/z(M+H)+calcd for 427.3934,C30H51O),1H-NMR(500MHz,CDCl3):δ0.76(s,3H),0.79(s,3H),0.83(s,3H),0.94(s,3H),0.97(s,3H),1.03(s,3H),1.68(s,3H),4.69(1H,d,J=1.98Hz,H-29a),4.57(1H,d,J=2.03Hz,H-29b)3.19(1H,dd,J=11.40,4.85Hz,3-H).
13C-NMR(125MHz,CDCl3):δ151.13,109.47,79.16,55.45,50.59,48.46,48.14,43.16,42.99,40.99,40.16,39.01,38.86,38.21,37.32,35.74,34.44,30.00,28.14,27.60,27.57,25.30,21.08,19.46,18.47,18.16,16.27,16.13,15.52,14.70.
化合物2:白色固体,C30H50O(HRESI-TOF-MS m/z(M+H)+calcd for 427.3934,C30H51O),1H-NMR(500MHz,CDCl3):δ0.80(m,9H),0.91(s,3H),0.95(s,3H),1.00(s,3H),1.01(s,3H),1.07(s,3H),5.13(1H,m,12-H)3.23(1H,dd,J=11.11,5.09Hz,3-H).
13C-NMR(125MHz,CDCl3):δ139.58,124.45,79.05,59.06,55.17,47.71,42.07,41.52,40.00,39.66,39.60,38.78,38.77,36.89,33.75,32.93,31.25,28.74,28.12,28.09,27.27,26.61,23.36,23.26,21.39,18.35,17.46,16.86,15.67,15.62.
化合物3:白色固体,C30H50O(HRESI-TOF-MS m/z(M+H)+calcd for 427.3934,C30H51O),1H-NMR(500MHz,CDCl3):δ0.79(s,3H),0.83(s,3H),0.87(s,6H),0.94(s,3H),0.97(s,3H),1.00(s,3H),1.07(s,3H),5.18(1H,m,12-H)3.22(1H,dd,J=11.22,4.59Hz,3-H).
13C-NMR(125MHz,CDCl3):δ145.34,121.85,79.18,55.28,47.75,47.34,46.95,41.84,39.91,38.91,38.70,37.27,37.07,34.86,33.49,32.77,32.63,31.23,28.55,28.23,27.35,27.05,26.28,26.14,23.83,23.66,18.51,16.93,15.73,15.65.
化合物4:白色固体,C30H50O(HRESI-TOF-MS m/z(M+H)+calcd for 427.3934,C30H51O),1H-NMR(500MHz,CDCl3):δ0.80(s,3H),0.82(s,3H),0.91(s,6H),0.93(s,3H),0.95(s,3H),0.98(s,3H),1.09(s,3H),5.53(1H,dd,J=8.19,3.26Hz,15-H)3.19(1H,dd,J=11.27,4.55Hz,3-H).
13C-NMR(125MHz,CDCl3):δ158.20,117.01,79.21,55.63,49.39,48.83,41.42,39.10,38.90,38.11,37.85,37.83,37.68,36.79,35.93,35.23,33.81,33.49,33.21,30.06,29.95,28.94,28.13,27.26,26.05,21.45,18.92,17.64,15.60,15.57.
化合物(5)的结构经LC-MS、1H NMR、13C NMR,HMBC测试后确定。
化合物5:白色固体;C47H76O17(HR ESI-TOF-MS m/z(M+Na)+calcd for 935.4975,C47H76O17Na);1H NMR(600MHz,pyridine-d5)δ5.60(1H,brt),3.31(1H,dd,J=3.82,11.55Hz),3.07(1H,s),1.22(3H,s),1.12(3H,s),0.86(3H,s),1.09(3H,s),1.44(3H,s),1.75(3H,s),1.12(3H,s),1.65(3H,d,J=5.98Hz),4.87(1H,d,J=5.30Hz),5.11(1H,d,J=7.80Hz),6.17(1H,brs)。
13C NMR(150MHz,pyridine-d5)δ38.88,27.5,88.66,39.94,56.45,18.99,33.87,40.71,48.09,37.38,24.33,128.36,140.29,42.46,29.68,26.76,48.67,54.97,73.07,42.72,26.99,39.4,28.46,17.12,15.97,17.38,25.06,181.13,27.3,17.55,105.09,74.36,82.48,68.52,65.17,102.26,72.76,72.87,74.28,70.38,18.93,105.01,75.32,78.58,71.81,78.92,62.89。
糖的比例及立体构型的确定
取化合物(5)3.0mg,1.25mL甲醇溶解后,加入250μL浓盐酸,80℃油浴回流4h,减压蒸馏干燥后,用水和氯仿交替萃取,所得水层悬干后得单糖混合物1.5mg,取200μg用160μL吡啶溶解,加入40μL 10mg/mL的L-半胱氨酸甲酯盐酸盐的吡啶溶液,60℃油浴1h后,再加入100μL含0.67μL邻甲苯异硫氰酸酯吡啶溶液,继续60℃油浴1h后取出,经氮吹干燥后,400μL甲醇复溶,取2μL进HR ESI-TOF-MS分析,糖的衍生物保留时间和标准糖衍生物进行比对,确定糖的绝对立体构型。标准糖衍生物的制备同上,其中由于标准D-鼠李糖无法购买得到,经查阅文献,将L-鼠李糖和D-半胱氨酸甲酯盐酸盐反应,其余条件不变,得到D-鼠李糖衍生物的保留时间。
化合物(5)的单糖衍生物保留时间(阿拉伯糖tR=6.900min,鼠李糖tR=7.493min和葡萄糖tR=6.713min)和标准糖衍生物L-半胱氨酸甲酯-D-阿拉伯糖(tR=6.780min),L-半胱氨酸甲酯-L-阿拉伯糖(tR=6.900min),L-半胱氨酸甲酯-D-葡萄糖(tR=6.720min),L-半胱氨酸甲酯-L-葡萄糖(tR=6.433min),L-半胱氨酸甲酯-L-鼠李糖(tR=7.513min),和D-半胱氨酸甲酯-L-鼠李糖(tR=6.140min)对比得到化合物(5)的单糖分别为L-阿拉伯糖,D-葡萄糖和L-鼠李糖。
另取化合物(5)的单糖混合物200μg,和200μL含100μg盐酸羟胺的吡啶溶液90℃油浴加热1h,取出冷却至室温,加4μL乙酸酐继续90℃油浴加热1h,氮吹干燥后,加400μL氯仿复溶,取0.2μL10:1分流进GC-MS分析,标准糖衍生物的制备同上,通过和标准糖的保留时间对比,确定糖的类型,再根据相应的GC-MS的峰面积确定糖的比例为1:1:1。
实施例3
中药苦丁茶活性组分(A),(b)-(f),(B)-(E),活性化合物(1)-(5)的生物活性。
在神经退行性病变动物模型中,研究发现NGF能阻止或减少神经元的退变,一定程度可阻止AD进展,具有促进神经生长和神经保护作用。由于PC12细胞具有神经细胞的一般特征,在NGF的作用下会停止分裂,长出突起,转化成神经元样细胞。因此,采用PC 12细胞作为有效的活性鉴定系统,筛选具有拟神经生长因子活性的化合物,有可能成为治疗阿尔兹海默症的有效药物。
实验方法:
1.PC 12细胞(大鼠肾上腺嗜铬瘤细胞株)的培养:在100mm的培养皿中,加入10mL含20×104个PC 12细胞DMEM培养基(其中含10%马血清、5%胎牛血清),两天后更换一次培养基,再过三天继代。继代时先用PBS(磷酸盐缓冲液)将细胞洗两次,再加入10mL PBS于培养皿中,在37℃,5%CO2的培养箱内静置10分钟后,取出吹洗,转移到15mL的一次性离心管,离心后血球计数板上计数。24孔细胞培养板每孔先加入1mL含血清的DMEM培养基,细胞计数后,每孔接2×104个细胞,CO2培养箱培养24小时后加样。
2.活性测试:以DMSO(二甲基亚砜)为阴性对照,NGF为阳性对照,将活性组分(A),(b)-(f),(B)-(E)及活性化合物(1)-(5)配置成不同浓度的DMSO溶液。用1mL含0.5%DMSO和不同浓度样品的DMEM溶液(不含血清)将24孔细胞板的每孔原培养基置换后,放入37℃,5%CO2的培养箱中培养。倒置显微镜下每隔24小时、连续4天观察细胞形态变化,记录细胞的神经突起分化率(神经突起长于胞体直径一倍的细胞数目与视野下总细胞数目的比值),每个视野下约100个细胞,随机选取3处,并统计作图分析。
3.实验结果:
在一定浓度下,加入苦丁茶酯层活性组分(A),(b)-(f),(B)-(E),活性化合物(1)-(5)后48小时,苦丁茶酯层活性组分(A),(b)-(f),(B)-(E),及活性化合物(1)-(5)均具有促PC 12细胞突起伸长的作用,显示出显著的拟神经生长因子活性。参见图1-4,以0.5%的DMSO作为阴性对照,样品活性组分(E)及化合物(4)和(5)的生物活性尤为突出。图1和图3中:0.5%DMSO为阴性对照;NGF:40ng/mL,阳性对照;***P<0.001。
图2和图4是加入苦丁茶酯层活性组分(A),(b)-(f),(B)-(E),活性化合物(1)-(5)后48小时PC 12细胞显微镜图片。从图2可以观察到苦丁茶酯层活性组分(A),(b)-(f)以及化合物(1)-(4)均能显著诱导PC 12细胞发生神经突起伸长的作用。图2中:a.阴性对照:0.5%DMSO;b.阳性对照:40ng/mL NGF;c.苦丁茶活性组分(A)100μg/mL;d.活性组分(b)30μg/mL;e.活性组分(c)30μg/mL;f.活性组分(d)10μg/mL;g.活性组分(e)10μg/mL;h.活性组分(f)10μg/mL;i.化合物(1)(1μM)。j.化合物(2)(1μM)k.化合物(3)(1μM)l.化合物(4)(1μM)。
图4中:a.阴性对照:0.5%DMSO;b.阳性对照:40ng/mL NGF;c.苦丁茶活性组分(A)100μg/mL;d.活性组分(B)30μg/mL;e.活性组分(C)10μg/mL;f.活性组分(D)10μg/mL;g.活性组分(E)10μg/mL;h.化合物(5)(3μM)。
Claims (10)
1.一种苦丁茶提取物的制备方法,其特征在于,通过以下步骤实现:
(1)粉碎和浸提:
取中药苦丁茶粉碎后,用甲醇溶液室温下震荡浸提3次,每次24小时,所得滤液经浓缩干燥后,用水和乙酸乙酯交替萃取分离,得到酯层活性组分(A);
(2)分离和纯化:
将活性组分(A)先经一次硅胶开口柱分离,以下溶剂系统均按体积比,经正己烷:乙酸乙酯洗脱,洗脱梯度:100:0,70:30,50:50,0:100,最后用甲醇冲洗,合并正己烷:乙酸乙酯70:30的洗脱流分,得到活性组分(b);合并甲醇的洗脱流分,得到活性组(B);
活性组分(b)经硅胶开口柱分离,正己烷:氯仿3:7洗脱,得活性组分(c),继续经硅胶开口柱分离,合并正己烷:氯仿5:5洗脱流分得活性组分(d),活性组分(d)经正己烷:氯仿7:3洗脱得活性组分(e);活性组分(e)用HPLC纯化,在tR=38min得到化合物(1)、tR=57min得化合物(2),得到活性组分(f);活性组分(f)继续用HPLC纯化,在tR=37min得到化合物(3)、在tR=35min得到化合物(4);
将活性组分(B)进一步用ODS开口柱分离,溶剂系统按体积比为甲醇:水=5:5,7:3,8:2,9:1,10:0,合并甲醇:水为8:2至9:1的洗脱流分,得到活性组分(C);活性组分(C)再次用硅胶开口柱分离,溶剂系统按体积比为氯仿:乙酸乙酯=95:5,90:10,80:20,70:30,60:40,50:50,40:60,30:70,20:80,90:10,100:0,合并甲醇:水为10:90至0:100的洗脱流分,得到活性组分(D);活性组分(D)用HPLC纯化,得到活性组分(E),继续用HPLC纯化,得到化合物(5)。
2.根据权利要求1所述的一种苦丁茶提取物的制备方法,其特征在于,活性组分(e)用HPLC纯化,色谱条件为:色谱柱流速3ml/min,检测波长210nm,流动相为99%乙腈水溶液等度洗脱。
3.根据权利要求1所述的一种苦丁茶提取物的制备方法,其特征在于,活性组分(f)继续用HPLC纯化,色谱条件为:色谱柱流速3ml/min,检测波长210nm,流动相为97%甲醇水溶液等度洗脱。
4.根据权利要求1所述的一种苦丁茶提取物的制备方法,其特征在于,活性组分(D)用HPLC纯化,色谱条件为:色谱柱流速3ml/min,检测波长210nm,流动相为45%乙腈水溶液等度洗脱30分钟。
5.根据权利要求1所述的一种苦丁茶提取物的制备方法,其特征在于,活性组分(D)用HPLC纯化,色谱条件为:色谱柱流速3ml/min,检测波长210nm,流动相为45%乙腈水溶液等度洗脱30分钟。
6.根据权利要求1所述的一种苦丁茶提取物的制备方法,其特征在于,活性组分(E)继续用HPLC纯化,色谱条件为:色谱柱流速3ml/min,检测波长210nm,流动相为63%甲醇水溶液等度洗脱50分钟。
7.根据权利要求1所述的一种苦丁茶提取物的制备方法,其特征在于,化合物(1)为羽扇豆醇,化合物(2)为α-香树脂醇,化合物(3)为β-香树脂醇,化合物(4)为蒲公英赛醇,化合物(5)为3β-O-D-葡萄糖(1→3)-[α-L-鼠李糖(1→3)]-α-L-阿拉伯糖-坡模酸。(3-O-β-D–glucopyranosyl-(1→3)-[α-L-rhamnopyranosyl-(1→2)]-α-L-arabinopyranosyl)-pomolic acid)。
8.根据权利要求1所述制备方法得到的苦丁茶提取物在制备预防或治疗阿尔兹海默症等神经退行性疾病的药物和保健食品中的应用,其特征在于,所述的苦丁茶提取物为活性组分(A),(b)-(f),(B)-(E),和化合物(1)-(5)。
9.根据权利要求8所述的应用,其特征在于,所述药物和保健食品由所述的苦丁茶提取物单独或组合,添加药学或食品可接受的载体或赋形剂制得,所述的苦丁茶提取物为活性组分(A),(b)-(f),(B)-(E),和化合物(1)-(5)。
10.根据权利要求8所述的应用,其特征在于,所述药物的给药途径为肠道或非肠道给药。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610601343.2A CN106236792B (zh) | 2016-07-26 | 2016-07-26 | 苦丁茶提取物的制备及抗阿尔兹海默症用途 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610601343.2A CN106236792B (zh) | 2016-07-26 | 2016-07-26 | 苦丁茶提取物的制备及抗阿尔兹海默症用途 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN106236792A true CN106236792A (zh) | 2016-12-21 |
CN106236792B CN106236792B (zh) | 2019-08-30 |
Family
ID=57604042
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201610601343.2A Active CN106236792B (zh) | 2016-07-26 | 2016-07-26 | 苦丁茶提取物的制备及抗阿尔兹海默症用途 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN106236792B (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109133356A (zh) * | 2018-09-12 | 2019-01-04 | 浙江海洋大学 | 一种利用滤食生物除藻的方法 |
CN113101293A (zh) * | 2021-04-19 | 2021-07-13 | 南通大学 | 熊果酸衍生物在制备治疗神经系统疾病药物中的应用 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH07151748A (ja) * | 1993-08-19 | 1995-06-16 | Eli Lilly & Co | 医薬スクリーニングにおけるアミリンの使用 |
CN1508149A (zh) * | 2002-12-18 | 2004-06-30 | 中南大学 | 从苦丁茶中提取总三萜酸及熊果酸、齐墩果酸的方法 |
CN101775061A (zh) * | 2009-09-15 | 2010-07-14 | 沈阳药科大学 | 一组苦丁茶皂苷类化合物 |
CN104435025A (zh) * | 2014-12-29 | 2015-03-25 | 广西梧州制药(集团)股份有限公司 | 苦丁茶在制备治疗阿尔兹海默病药物方面的新用途 |
CN105030914A (zh) * | 2015-07-14 | 2015-11-11 | 四川大学 | 粗壮女贞苦丁茶提取物在α-葡萄糖苷酶抑制剂的应用 |
-
2016
- 2016-07-26 CN CN201610601343.2A patent/CN106236792B/zh active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH07151748A (ja) * | 1993-08-19 | 1995-06-16 | Eli Lilly & Co | 医薬スクリーニングにおけるアミリンの使用 |
CN1508149A (zh) * | 2002-12-18 | 2004-06-30 | 中南大学 | 从苦丁茶中提取总三萜酸及熊果酸、齐墩果酸的方法 |
CN101775061A (zh) * | 2009-09-15 | 2010-07-14 | 沈阳药科大学 | 一组苦丁茶皂苷类化合物 |
CN104435025A (zh) * | 2014-12-29 | 2015-03-25 | 广西梧州制药(集团)股份有限公司 | 苦丁茶在制备治疗阿尔兹海默病药物方面的新用途 |
CN105030914A (zh) * | 2015-07-14 | 2015-11-11 | 四川大学 | 粗壮女贞苦丁茶提取物在α-葡萄糖苷酶抑制剂的应用 |
Non-Patent Citations (2)
Title |
---|
李国胜,严亮: "苦丁茶提取物的抗氧化活性研究", 《轻工科技》 * |
胡婷: "苦丁茶中有效成分的分离纯化、鉴定及其活性研究", 《中国优秀硕士学位论文全文数据库工程科技Ⅰ辑》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109133356A (zh) * | 2018-09-12 | 2019-01-04 | 浙江海洋大学 | 一种利用滤食生物除藻的方法 |
CN113101293A (zh) * | 2021-04-19 | 2021-07-13 | 南通大学 | 熊果酸衍生物在制备治疗神经系统疾病药物中的应用 |
CN113101293B (zh) * | 2021-04-19 | 2022-02-01 | 南通大学 | 熊果酸衍生物在制备治疗神经系统疾病药物中的应用 |
Also Published As
Publication number | Publication date |
---|---|
CN106236792B (zh) | 2019-08-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN101486743B (zh) | 具有抗老年痴呆症作用的新环烯醚萜类化合物 | |
CN103059043A (zh) | 伸筋草碱a-c、其制法和其药物组合物与用途 | |
CN103191174B (zh) | 杜仲化学成分作为血管保护剂的新用途 | |
CN105294623B (zh) | 一种倍半萜内酯类化合物、其制备方法以及应用 | |
CN101214253B (zh) | 知母皂苷b-ⅱ用于制备抗抑郁产品的用途 | |
CN101647850A (zh) | 杜仲化学成分作为植物雌激素的新用途 | |
CN107556362A (zh) | 葫芦烷型三萜化合物的提取方法及抗阿尔兹海默症医药用途 | |
CN106236792A (zh) | 苦丁茶提取物的制备及抗阿尔兹海默症用途 | |
CN103360456B (zh) | 三萜类化合物及制备和应用 | |
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 | |
CN102935100A (zh) | 一种大花罗布麻叶总黄酮的制备方法和用途 | |
CN111647003B (zh) | 三环氧六氢色酮a及其药物组合物和其应用 | |
CN111848565A (zh) | 单萜基双香豆素类化合物、药物组合物及其制备方法和用途 | |
CN103288914B (zh) | 中药饿蚂蝗提取物的制备及抗老年性痴呆药物用途 | |
CN103641713B (zh) | 甘油单酯衍生物的制备方法及应用 | |
CN101774922B (zh) | 2,3-二羟基苯甲酸酯类化合物及制备和应用 | |
CN102188477B (zh) | 龙胆提取物活性组分的制备及应用 | |
CN101612184B (zh) | 多舌飞蓬提取物、含该提取物的组合物及制备方法和用途 | |
CN104224796B (zh) | 齐墩果烷型三萜类酯衍生物抗神经退行性药物用途 | |
CN111529515B (zh) | 12,15-二氧-α-蛇床烯在制药中的应用 | |
CN103494813A (zh) | 盐酸去亚甲基小檗碱在制备预防和/或治疗急慢性酒精性肝病药物中的应用 | |
CN103342730B (zh) | 中药饿蚂蝗提取物的制备方法及其抗衰老用途 | |
CN102250197A (zh) | 一种麦冬总甾体皂苷提取物的制备方法及应用 | |
CN101461803B (zh) | 葡萄内酯的用途 | |
CN1686254A (zh) | 苍耳子总苷提取物用于制备抗炎性反应产品的用途 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |