CN101347502A - Effective component of selfheal and preparation and use thereof - Google Patents
Effective component of selfheal and preparation and use thereof Download PDFInfo
- Publication number
- CN101347502A CN101347502A CNA2007101232735A CN200710123273A CN101347502A CN 101347502 A CN101347502 A CN 101347502A CN A2007101232735 A CNA2007101232735 A CN A2007101232735A CN 200710123273 A CN200710123273 A CN 200710123273A CN 101347502 A CN101347502 A CN 101347502A
- Authority
- CN
- China
- Prior art keywords
- active component
- ethyl acetate
- eluent
- mobile phase
- obtains
- 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
- 238000002360 preparation method Methods 0.000 title claims abstract description 41
- 244000299788 selfheal Species 0.000 title 1
- 235000008113 selfheal Nutrition 0.000 title 1
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 108
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 80
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 39
- 239000000203 mixture Substances 0.000 claims abstract description 23
- 239000000284 extract Substances 0.000 claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000010828 elution Methods 0.000 claims abstract description 9
- 239000002904 solvent Substances 0.000 claims abstract description 7
- 238000004440 column chromatography Methods 0.000 claims abstract description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 36
- 239000003480 eluent Substances 0.000 claims description 32
- 235000019441 ethanol Nutrition 0.000 claims description 27
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 22
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 22
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 20
- 238000004262 preparative liquid chromatography Methods 0.000 claims description 16
- 239000000741 silica gel Substances 0.000 claims description 16
- 229910002027 silica gel Inorganic materials 0.000 claims description 16
- 229960001866 silicon dioxide Drugs 0.000 claims description 16
- 230000008859 change Effects 0.000 claims description 12
- 238000001035 drying Methods 0.000 claims description 11
- 239000003208 petroleum Substances 0.000 claims description 11
- 239000012141 concentrate Substances 0.000 claims description 10
- -1 reflux Substances 0.000 claims description 9
- 238000000926 separation method Methods 0.000 claims description 8
- 238000010992 reflux Methods 0.000 claims description 7
- 239000000463 material Substances 0.000 claims description 5
- 239000012567 medical material Substances 0.000 claims description 5
- 238000002953 preparative HPLC Methods 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 4
- 230000001476 alcoholic effect Effects 0.000 claims description 4
- 239000000706 filtrate Substances 0.000 claims description 4
- 239000008194 pharmaceutical composition Substances 0.000 claims description 4
- 238000010298 pulverizing process Methods 0.000 claims description 3
- 244000179560 Prunella vulgaris Species 0.000 abstract description 4
- 235000010674 Prunella vulgaris Nutrition 0.000 abstract 3
- 239000007791 liquid phase Substances 0.000 abstract 1
- 239000012071 phase Substances 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 68
- 239000012531 culture fluid Substances 0.000 description 36
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 35
- 239000003814 drug Substances 0.000 description 35
- 239000000243 solution Substances 0.000 description 28
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 24
- 239000007788 liquid Substances 0.000 description 24
- 238000002156 mixing Methods 0.000 description 22
- 230000001629 suppression Effects 0.000 description 19
- 229960000583 acetic acid Drugs 0.000 description 15
- 238000010790 dilution Methods 0.000 description 15
- 239000012895 dilution Substances 0.000 description 15
- 210000004881 tumor cell Anatomy 0.000 description 12
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 10
- 239000012490 blank solution Substances 0.000 description 10
- 239000012362 glacial acetic acid Substances 0.000 description 10
- 230000004083 survival effect Effects 0.000 description 10
- 206010028980 Neoplasm Diseases 0.000 description 9
- 238000004113 cell culture Methods 0.000 description 8
- 229940079593 drug Drugs 0.000 description 8
- 239000006228 supernatant Substances 0.000 description 8
- 239000002552 dosage form Substances 0.000 description 7
- 238000005303 weighing Methods 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 239000013543 active substance Substances 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 6
- 239000003795 chemical substances by application Substances 0.000 description 6
- 235000008504 concentrate Nutrition 0.000 description 6
- 239000000725 suspension Substances 0.000 description 6
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 6
- 235000011054 acetic acid Nutrition 0.000 description 5
- 230000009514 concussion Effects 0.000 description 5
- 230000000857 drug effect Effects 0.000 description 5
- 238000001704 evaporation Methods 0.000 description 5
- 230000008020 evaporation Effects 0.000 description 5
- 239000003292 glue Substances 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 239000013642 negative control Substances 0.000 description 5
- 210000002826 placenta Anatomy 0.000 description 5
- 239000013641 positive control Substances 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 5
- 235000017557 sodium bicarbonate Nutrition 0.000 description 5
- 239000003826 tablet Substances 0.000 description 5
- 238000011282 treatment Methods 0.000 description 5
- NWXMGUDVXFXRIG-WESIUVDSSA-N (4s,4as,5as,6s,12ar)-4-(dimethylamino)-1,6,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide Chemical compound C1=CC=C2[C@](O)(C)[C@H]3C[C@H]4[C@H](N(C)C)C(=O)C(C(N)=O)=C(O)[C@@]4(O)C(=O)C3=C(O)C2=C1O NWXMGUDVXFXRIG-WESIUVDSSA-N 0.000 description 4
- 229930195573 Amycin Natural products 0.000 description 4
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 4
- 229930195725 Mannitol Natural products 0.000 description 4
- 238000007605 air drying Methods 0.000 description 4
- 239000003797 essential amino acid Substances 0.000 description 4
- 235000020776 essential amino acid Nutrition 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000012737 fresh medium 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
- 239000000594 mannitol Substances 0.000 description 4
- 235000010355 mannitol Nutrition 0.000 description 4
- 239000002547 new drug Substances 0.000 description 4
- 239000003921 oil Substances 0.000 description 4
- 235000019198 oils Nutrition 0.000 description 4
- 238000011017 operating method Methods 0.000 description 4
- 239000006187 pill Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 230000001093 anti-cancer Effects 0.000 description 3
- 230000000259 anti-tumor effect Effects 0.000 description 3
- 244000309466 calf Species 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- 238000004737 colorimetric analysis Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000000945 filler Substances 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 239000008187 granular material Substances 0.000 description 3
- 239000011259 mixed solution Substances 0.000 description 3
- 239000002674 ointment Substances 0.000 description 3
- 238000012856 packing Methods 0.000 description 3
- 230000000144 pharmacologic effect Effects 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-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
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- 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 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 206010018498 Goitre Diseases 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 241001299553 Ilex chinensis Species 0.000 description 2
- 241001100935 Ilex purpurea Species 0.000 description 2
- 235000003366 Ilex purpurea Nutrition 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000002421 anti-septic effect Effects 0.000 description 2
- SHWNNYZBHZIQQV-UHFFFAOYSA-L calcium;disodium;2-[2-[bis(carboxylatomethyl)azaniumyl]ethyl-(carboxylatomethyl)azaniumyl]acetate Chemical compound [Na+].[Na+].[Ca+2].[O-]C(=O)C[NH+](CC([O-])=O)CC[NH+](CC([O-])=O)CC([O-])=O SHWNNYZBHZIQQV-UHFFFAOYSA-L 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 238000004043 dyeing Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 150000002148 esters Chemical class 0.000 description 2
- 238000000105 evaporative light scattering detection Methods 0.000 description 2
- 235000013355 food flavoring agent Nutrition 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 201000003872 goiter Diseases 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 239000000314 lubricant Substances 0.000 description 2
- 201000007227 lymph node tuberculosis Diseases 0.000 description 2
- 235000019359 magnesium stearate Nutrition 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 2
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 2
- 235000019345 sodium thiosulphate Nutrition 0.000 description 2
- 239000000600 sorbitol Substances 0.000 description 2
- 235000010356 sorbitol Nutrition 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 229940032147 starch Drugs 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- QIJRTFXNRTXDIP-UHFFFAOYSA-N (1-carboxy-2-sulfanylethyl)azanium;chloride;hydrate Chemical compound O.Cl.SCC(N)C(O)=O QIJRTFXNRTXDIP-UHFFFAOYSA-N 0.000 description 1
- 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 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- CYDQOEWLBCCFJZ-UHFFFAOYSA-N 4-(4-fluorophenyl)oxane-4-carboxylic acid Chemical compound C=1C=C(F)C=CC=1C1(C(=O)O)CCOCC1 CYDQOEWLBCCFJZ-UHFFFAOYSA-N 0.000 description 1
- LXAHHHIGZXPRKQ-UHFFFAOYSA-N 5-fluoro-2-methylpyridine Chemical compound CC1=CC=C(F)C=N1 LXAHHHIGZXPRKQ-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 235000019489 Almond oil Nutrition 0.000 description 1
- 239000005995 Aluminium silicate Substances 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 208000017897 Carcinoma of esophagus Diseases 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 1
- 206010051625 Conjunctival hyperaemia Diseases 0.000 description 1
- 206010010726 Conjunctival oedema Diseases 0.000 description 1
- 229920000858 Cyclodextrin Polymers 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
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 241000201295 Euphrasia Species 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 206010019233 Headaches Diseases 0.000 description 1
- 239000004354 Hydroxyethyl cellulose Substances 0.000 description 1
- 229920000663 Hydroxyethyl cellulose Polymers 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- 208000002454 Nasopharyngeal Carcinoma Diseases 0.000 description 1
- 206010061306 Nasopharyngeal cancer Diseases 0.000 description 1
- 241001597008 Nomeidae Species 0.000 description 1
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Natural products OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 229920002584 Polyethylene Glycol 6000 Polymers 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 239000004141 Sodium laurylsulphate Substances 0.000 description 1
- NWGKJDSIEKMTRX-AAZCQSIUSA-N Sorbitan monooleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O NWGKJDSIEKMTRX-AAZCQSIUSA-N 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 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 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000001785 acacia senegal l. willd gum Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229910000288 alkali metal carbonate Inorganic materials 0.000 description 1
- 150000008041 alkali metal carbonates Chemical class 0.000 description 1
- 239000008168 almond oil Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 235000012211 aluminium silicate Nutrition 0.000 description 1
- CEGOLXSVJUTHNZ-UHFFFAOYSA-K aluminium tristearate Chemical compound [Al+3].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O CEGOLXSVJUTHNZ-UHFFFAOYSA-K 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 239000003005 anticarcinogenic agent Substances 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 239000008365 aqueous carrier Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 201000008275 breast carcinoma Diseases 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- NKWPZUCBCARRDP-UHFFFAOYSA-L calcium bicarbonate Chemical compound [Ca+2].OC([O-])=O.OC([O-])=O NKWPZUCBCARRDP-UHFFFAOYSA-L 0.000 description 1
- 229910000020 calcium bicarbonate Inorganic materials 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- CJZGTCYPCWQAJB-UHFFFAOYSA-L calcium stearate Chemical compound [Ca+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O CJZGTCYPCWQAJB-UHFFFAOYSA-L 0.000 description 1
- 239000008116 calcium stearate Substances 0.000 description 1
- 235000013539 calcium stearate Nutrition 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 239000003240 coconut oil Substances 0.000 description 1
- 235000019864 coconut oil Nutrition 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 229960001305 cysteine hydrochloride Drugs 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- FPAFDBFIGPHWGO-UHFFFAOYSA-N dioxosilane;oxomagnesium;hydrate Chemical compound O.[Mg]=O.[Mg]=O.[Mg]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O FPAFDBFIGPHWGO-UHFFFAOYSA-N 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- 239000008298 dragée Substances 0.000 description 1
- 239000006196 drop Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000008157 edible vegetable oil Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 239000002662 enteric coated tablet Substances 0.000 description 1
- 201000005619 esophageal carcinoma Diseases 0.000 description 1
- BEFDCLMNVWHSGT-UHFFFAOYSA-N ethenylcyclopentane Chemical compound C=CC1CCCC1 BEFDCLMNVWHSGT-UHFFFAOYSA-N 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000019197 fats Nutrition 0.000 description 1
- 239000007941 film coated tablet Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 210000000232 gallbladder Anatomy 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001879 gelation Methods 0.000 description 1
- 239000007902 hard capsule Substances 0.000 description 1
- 231100000869 headache Toxicity 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 235000011167 hydrochloric acid Nutrition 0.000 description 1
- 238000005984 hydrogenation reaction Methods 0.000 description 1
- 235000019447 hydroxyethyl cellulose Nutrition 0.000 description 1
- 229940071826 hydroxyethyl cellulose Drugs 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- FZWBNHMXJMCXLU-BLAUPYHCSA-N isomaltotriose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@@H](OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O)O1 FZWBNHMXJMCXLU-BLAUPYHCSA-N 0.000 description 1
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 229940057995 liquid paraffin Drugs 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000003589 local anesthetic agent Substances 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- 235000019426 modified starch Nutrition 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 201000011216 nasopharynx carcinoma Diseases 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 229960003742 phenol Drugs 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 235000011007 phosphoric acid Nutrition 0.000 description 1
- 239000011505 plaster Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229940093429 polyethylene glycol 6000 Drugs 0.000 description 1
- 239000001103 potassium chloride Substances 0.000 description 1
- 235000011164 potassium chloride Nutrition 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 1
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 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
- 238000003908 quality control method Methods 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 150000003376 silicon Chemical class 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000002639 sodium chloride Nutrition 0.000 description 1
- HRZFUMHJMZEROT-UHFFFAOYSA-L sodium disulfite Chemical compound [Na+].[Na+].[O-]S(=O)S([O-])(=O)=O HRZFUMHJMZEROT-UHFFFAOYSA-L 0.000 description 1
- 239000001540 sodium lactate Substances 0.000 description 1
- 229940005581 sodium lactate Drugs 0.000 description 1
- 235000011088 sodium lactate Nutrition 0.000 description 1
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 1
- 235000010262 sodium metabisulphite Nutrition 0.000 description 1
- 229940080313 sodium starch Drugs 0.000 description 1
- 235000010265 sodium sulphite Nutrition 0.000 description 1
- 239000007901 soft capsule Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 235000010199 sorbic acid Nutrition 0.000 description 1
- 239000004334 sorbic acid Substances 0.000 description 1
- 229940075582 sorbic acid Drugs 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000007940 sugar coated tablet Substances 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
Images
Landscapes
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to an effective component of prunella vulgaris, a preparation method and application thereof; the preparation process of the effective component of prunella vulgaris in the invention includes the following steps: step 1, the mixture of ethyl acetate and ethanol is used as solvent for extracting prunella vulgaris; step 2, the extract is processed with chromatographic column chromatography to obtain eluate; step3, the eluate obtained by elution in the preparation of liquid phase chromatographic gradient is adopted; the mobile phase is water and acetonitrile, and the eluate of 24.0-28.0 min or 28.0-32.0 min is collected to obtain the effective component.
Description
Technical field
The present invention relates to a kind of Chinese medicine extract for the treatment of tumor disease, relate in particular to the active component that from Spica Prunellae, extracts, preparation and preparation method thereof and purposes.
Background technology
Tumor is a kind of commonly encountered diseases, frequently-occurring disease, and wherein malignant tumor is the most serious class disease of present harm humans health.At present in the industry to the treatment of malignant tumor mainly still based on operation, radiotherapy, chemotherapy, but many chemical anticarcinogenic drugs often involve normal cell when acting on target cell, cause serious side reaction.The genetoxic of plant amedica is not obvious, and Chinese herbal medicine is having special advantages and wide application prospect aspect the anticancer mutation, and Chinese medicine also plays the effect that can not be ignored in to the auxiliary treatment of tumor.Paclitaxel promptly is the good anticancer active native compound that has that typically obtains from plant, now has been developed as antitumor drug.The most serious tumor of China's hazardness is pulmonary carcinoma, nasopharyngeal carcinoma, the esophageal carcinoma, gastric cancer, colorectal cancer, hepatocarcinoma, breast carcinoma, cervical cancer, leukemia and lymphoma etc. at present.Particularly the incidence rate of hepatocarcinoma increases in recent years to some extent.Significant, the etiology of these tumors, pathogenesis and control thereof are the emphasis of China's tumor research.Cancer therapy drug and the anticancer ancillary drug of seeking high-efficiency low-toxicity are the important contents of current tumor research.
China's medicinal organism resource is very abundant, and its biological active substances is research and finds new drug guide chemicals, the natural treasure-house of developing new drug.At present, China extracts active substance from natural product, be used to be developed to treatment tumor disease, safety is good, toxicity is low new drug also seldom, extracts active substance from natural product, be developed to new drug, have significant application value and wide development prospect with antitumor curative effect.
Spica Prunellae, another name: withered, big headdress flower of iron wire summer, Pemisetum flaccidum Griseb, June are done, wooden club is careless, medicinal part: fruit ear, its meridian distribution of property and flavor: cold; Bitter, hot; Return liver, gallbladder meridian, its function cure mainly for: relieve inflammation or internal heat, make eye bright, eliminating stagnation, the detumescence.Be used for conjunctival congestion and swelling pain, order pearl nyctalgia, it is dizzy to have a headache, scrofula, goiter swells and ache; Thyromegaly, tuberculous lymphadenitis, cyclomastopathy, hypertension.
Summary of the invention:
The object of the present invention is to provide the active component of two kinds of Spica Prunellaes.
Another object of the present invention is to provide the preparation method of above-mentioned Spica Prunellae active component.
The present invention also provides the preparation that contains above-mentioned Spica Prunellae active component and the purposes of this component.
Spica Prunellae active component of the present invention, its preparation process may further comprise the steps:
Step 1: Spica Prunellae is extracted as solvent with ethyl acetate and alcohol mixture,
Step 2: extracting solution gets eluent through column chromatography;
Step 3: the eluent that obtains with the preparative liquid chromatography gradient elution, mobile phase is water and acetonitrile, collect 24.0-28.0 minute eluent and obtain active component 1 (or being called C06), or 28.0-32.0 minute collection eluent obtains active component 2 (or being called C07).These two kinds of components are active component of the present invention.
Wherein ethyl acetate described in the step 1 and alcoholic acid mixture, both ratios are ethyl acetate: ethanol=1-5: 1-5, are preferably ethyl acetate: ethanol=1-2: 1-2 most preferably is ethyl acetate: ethanol=1: 1.
In the described step, step 1 is specially: getting the Spica Prunellae medical material, is solvent with ethyl acetate: ethanol=1-5: 1-5, and reflux, extract, is separated extracting solution with medicinal residues, and the extracting solution that obtains after the separation is an extract 1,
Step 2 is specially: extract 1 is crossed normal phase silicagel column, and use petroleum ether: ethyl acetate=47-53 earlier: 1 as the mobile phase eluting, changes chloroform then: methanol 8-13: 1 as mobile phase, gets eluent,
Step 3 is specially: continue to separate the eluent that obtains with preparative liquid chromatography, mobile phase is water-A and acetonitrile-B, carries out gradient elution, and flow velocity is 9-11ml/min, column temperature is a room temperature, collects 24.0-28.0 minute or 28.0-32.0 minute eluent obtains active component 1 and 2.
The program of gradient elution described in the step 3 is as follows: table 1 gradient table
Flow velocity is 10ml/min, and column temperature is a room temperature; Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 24.0-28.0 minute or 28.0-32.0 minute, and solution obtains active component 1 and 2 behind concentrate drying.
The preferred Spica Prunellae active component of the present invention preparation method, comprise the following steps: and to add ethyl acetate and ethanol (1: 0.8-1.2) after the Spica Prunellae pulverizing medicinal materials, reflux 0.8-1.2 hour, extract 1-3 time, merging filtrate gets extracting solution, extracting solution is condensed into extractum, and itself and silica gel mixed sample, with normal phase silicagel column it is separated, at first use petroleum ether and ethyl acetate (47-53:, get eluent I 1) as mobile phase, abandon it, change chloroform and methanol (8-13: 1) as mobile phase, get eluent II, will get sample behind the eluent II concentrate drying then; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is a preparative column, and mobile phase is water and acetonitrile, and gradient elution, flow velocity are 9-11ml/min, and column temperature is a room temperature.
The most preferred Spica Prunellae active component of the present invention preparation method comprises the following steps: and will add ethyl acetate and ethanol (1: 1) after the Spica Prunellae pulverizing medicinal materials that reflux 1 hour is extracted 2 times, and merging filtrate gets extracting solution; Extracting solution is condensed into extractum, and itself and silica gel mixed sample, with normal phase silicagel column it is separated, at first use petroleum ether and ethyl acetate (50: 1) as mobile phase, get eluent I, abandon it, change chloroform and methanol (10: 1) then as mobile phase, get eluent II, will get sample behind the eluent II concentrate drying; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is preparative column (Zorbax SB-C18; 21.2mm * 250mm), mobile phase is water (A) and acetonitrile (B), the gradient elution program is as follows:
| Time(min) | A(％) | B(％) |
| 0 | 80 | 20 |
| 4 | 80 | 20 |
| 19 | 50 | 50 |
| 54 | 5 | 95 |
| 64 | 5 | 95 |
Flow velocity is 10ml/min, and column temperature is a room temperature; Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 24.0-28.0 minute or 28.0-32.0 minute, and solution obtains active component 1 and 2 behind concentrate drying.
Active component composition such as the following table that collect 24.0-28.0 of the present invention minute or 28.0-32.0 minute
Table 2 component list
The present invention also provides the pharmaceutical composition that is prepared into as active constituents of medicine with Chinese medicine active component of the present invention, and pharmaceutical composition of the present invention comprises active component, and said composition can also add the medicine acceptable carrier as required.
Compositions of the present invention is the pharmaceutical dosage forms of unit dose, and described unit dosage form is meant the unit of preparation, as every of tablet, and capsular every capsules, every bottle of oral liquid, every bag of granule etc.
Compositions of the present invention active component wherein, its shared percentage by weight in preparation can be 0.1-99.9%, all the other are the medicine acceptable carrier.
Compositions of the present invention obtains by above-mentioned active component and medicine acceptable carrier are mixed with.
Compositions of the present invention, its pharmaceutical dosage forms can be any pharmaceutically useful dosage form, and these dosage forms comprise: tablet, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard capsule, soft capsule, oral liquid, suck agent, granule, electuary, pill, powder, unguentum, sublimed preparation, suspensoid, powder, solution, injection, suppository, ointment, plaster, cream, spray, drop, patch.Preparation of the present invention, peroral dosage form preferably, as: capsule, tablet, oral liquid, granule, pill, powder, sublimed preparation, unguentum etc.
Compositions of the present invention, the preparation of its oral administration can contain excipient commonly used, such as binding agent, filler, diluent, tablet agent, lubricant, disintegrating agent, coloring agent, flavoring agent and wetting agent, can carry out coating to tablet in case of necessity.
The filler that is suitable for comprises cellulose, mannitol, lactose and other similar filler.Suitable disintegrating agent comprises starch, polyvinylpyrrolidone and starch derivatives, for example sodium starch glycollate.Suitable lubricant comprises, for example magnesium stearate.The acceptable wetting agent of appropriate drug comprises sodium lauryl sulphate.
Can fill by mixing, the method that tabletting etc. are commonly used prepares solid oral composition.Mix repeatedly active substance is distributed in those compositionss of a large amount of filleies of whole use.
The form of oral liquid for example can be aqueous or oily suspensions, solution, Emulsion, syrup or elixir, perhaps can be a kind of available water before use or other suitable composite dry products of carrier.This liquid preparation can contain conventional additive, such as suspending agent, for example sorbitol, syrup, methylcellulose, gelatin, hydroxyethyl-cellulose, carboxymethyl cellulose, aluminium stearate gel or hydrogenation edible fat, emulsifying agent, for example lecithin, anhydro sorbitol monooleate or arabic gum; Non-aqueous carrier (they can comprise edible oil), for example almond oil, fractionated coconut oil, such as oily ester, propylene glycol or the ethanol of the ester of glycerol; Antiseptic, for example para hydroxybenzene methyl ester or propyl p-hydroxybenzoate or sorbic acid, and if desired, can contain conventional flavouring agent or coloring agent.
For injection, the liquid unit dosage forms of preparation contains active substance of the present invention and sterile carrier.According to carrier and concentration, this chemical compound can be suspended or dissolving.The preparation of solution is normally by being dissolved in active substance in a kind of carrier filter-sterilized before it is packed into a kind of suitable bottle or ampoule, sealing then.For example a kind of local anesthetic of adjuvant, antiseptic and buffer agent also can be dissolved in this carrier.In order to improve its stability, can be after the bottle of packing into that this compositions is freezing, and under vacuum, water is removed.
Compositions of the present invention, when being prepared into medicament, optionally add suitable medicine acceptable carrier, described medicine acceptable carrier is selected from: mannitol, sorbitol, sodium pyrosulfite, sodium sulfite, sodium thiosulfate, cysteine hydrochloride, TGA, methionine, vitamin C, the EDTA disodium, EDTA calcium sodium, the alkali-metal carbonate of monovalence, acetate, phosphate or its aqueous solution, hydrochloric acid, acetic acid, sulphuric acid, phosphoric acid, aminoacid, sodium chloride, potassium chloride, sodium lactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch, sucrose, lactose, mannitol, silicon derivative, cellulose and derivant thereof, alginate, gelatin, polyvinylpyrrolidone, glycerol, soil temperature 80, agar, calcium carbonate, calcium bicarbonate, surfactant, Polyethylene Glycol, cyclodextrin, beta-schardinger dextrin-, the phospholipid material, Kaolin, Pulvis Talci, calcium stearate, magnesium stearate etc.
Compositions of the present invention is determined usage and dosage according to patient's situation in use, but obeys every day three times, each 1-20 agent, as: 1-20 bag or grain or sheet.
The present invention also provides the application at anti-tumor aspect of Chinese medicine active component of the present invention and pharmaceutical composition.Below be the data of pharmacological evaluation:
Screening active ingredients
On cellular level, detect the growth inhibited effect of active component to kinds of tumor cells.
Cell strain: HL-60 tumor cell
Sample preparation: according to the weight of the medicine of institute's weighing, add the DMSO dissolving of respective volume, concentration is about 50mg/mL.The concussion dissolving, if there are a large amount of drops to glue wall, can be suitably centrifugal.Can store-20 ℃.
Cell culture: use RPMI 1640 (Gibco) [adding the 2g/L sodium bicarbonate] 90%, hyclone (Ilex purpurea Hassk.[I.chinensis Sims) 10% Mixed culture HL60 cell, density need be lower than 106/mL.
Experimental technique:
1 kind of plate:
(1) calculates and to need cell total amount NT=ρ cellmL-1 * VT (ρ=2 * 104/mL), wherein VT=0.1mL * hole count+cell groove surplus.
(2) suspension cell on the piping and druming culture bottle wall adds in the centrifuge tube.Get 10 μ L, add the blue dilution of 10 μ L Placenta Hominiss, viable count sum NL on the count plate, then the cell number N in the centrifuge tube is: NL/4 * 104 * 2 * V, wherein V is the liquor capacity in the centrifugal preceding centrifuge tube.
It is (3) centrifugal that (900rad/min 10min), inhales and removes supernatant, adds culture fluid V1mL, and piping and druming makes the cell mixing, draws V2mL and joins in the cell groove, makes V2=NT/N * V1.
(4) add the culture fluid of VT-V2mL again in the cell groove, with volley of rifle fire piping and druming, mixing, get this liquid, every hole adds 150 μ L.
(5) choose 4 holes behind the kind plate and add 200 μ L culture fluid as blank, the residue hole adds 200 μ L PBS, to reduce the evaporation of culture fluid.
A) dosing
The culture fluid that adds 220 μ L/ holes in 96 new orifice plates will be drawn 0.88 μ L medicinal liquid and add mixing, dilute 250 times, and the medicine that above-mentioned dilution is good has every hole, hole of cell to add 50 μ L to kind, and this moment, drug dilution was 1000 times, and promptly final concentration is 50ng/mL.Hatch 48h.Each concentration is established 4 parallel multiple holes, and every plate is established negative control group (add blank solution in the cell, blank solution is joined method---add the culture fluid of 200 μ L, will draw 0.88 μ LDMSO and add mixing), reaches positive controls (cisplatin final concentration 4 μ g/mL).
B) SRB dyeing
After cell culture finishes, take out culture plate, every hole adds trichloroacetic acid (TCA) the 100 μ L fixed cells of 40% (mass/volume), and room temperature is placed 5min, places 1h in 4 ℃ of refrigerators.Deionized water wash 5 times of each hole of culture plate are to remove TCA.Behind air drying, every hole adds 0.4% SRB100 μ L (dissolving of 1% chromatographically pure acetic acid), place 20min under the room temperature, discard in each hole and wash 5 times with 1% acetic acid behind the liquid, remove unconjugated dyestuff, with 10mmol/L Tris150 μ L/ hole dissolving, 5min vibrates behind the air drying, use microplate reader (ELx 800) to measure, used wavelength is 490nm.
The calculating suppression ratio of 4 suppression ratio is calculated as follows:
Drug effect the results are shown in Table 3.
Table 3HL-60 inhibition rate of tumor cell result
| C06 | Negative | Blank | Positive | |
| Average cell survival number | 0.189 | 0.725 | 0.064 | 0.15 |
| Suppression ratio (%) | 81.05 | 0 | 100 | 86.99 |
| RSD(％) | 4.033 | 2.272 | 7.974 | 1.440 |
| C07 | Negative | Blank | Positive | |
| Average cell survival number | 0.184 | 0.725 | 0.064 | 0.15 |
| Suppression ratio (%) | 81.92 | 0 | 100 | 86.99 |
| RSD(％) | 3.285 | 2.272 | 7.974 | 1.440 |
Cell strain: K562 tumor cell
Sample preparation: according to the weight of the medicine of institute's weighing, add the DMSO dissolving of respective volume, concentration is about 50mg/mL.The concussion dissolving, if there are a large amount of drops to glue wall, can be suitably centrifugal.Can store-20 ℃.
Cell culture: use RPMI 1640 (Gibco) [adding the 2g/L sodium bicarbonate] 90%, calf serum (match is happy) 10%, non essential amino acid (Gibco) 1% Mixed culture K 562 cells.
Experimental technique:
1 kind of plate:
(1) calculates and to need cell total amount NT=ρ cellmL-1 * VT (ρ=8 * 103/mL), wherein VT=0.1mL * hole count+cell groove surplus.
(2) suspension cell on the piping and druming culture bottle wall adds in the centrifuge tube.Get 10 μ L, add the blue dilution of 10 μ L Placenta Hominiss, viable count sum NL on the count plate, then the cell number N in the centrifuge tube is: NL/4 * 104 * 2 * V, wherein V is the liquor capacity in the centrifugal preceding centrifuge tube.
It is (3) centrifugal that (900rad/min 10min), inhales and removes supernatant, adds culture fluid V1mL, and piping and druming makes the cell mixing, draws V2mL and joins in the cell groove, makes V2=NT/N * V1.
(4) add the culture fluid of VT-V2mL again in the cell groove, with volley of rifle fire piping and druming, mixing, get this liquid, every hole adds 100 μ L, hatches 24h.
(5) choose 4 holes behind the kind plate and add culture fluid as blank, the residue hole adds 100 μ L PBS, to reduce the evaporation of culture fluid.
2 dosings:
(1) 96 orifice plate changes liquid, and every hole adds fresh medium 150 μ L.
(2) culture fluid in adding 220 μ L/ holes in 96 new orifice plates, to draw 0.88 μ L medicinal liquid and add mixing, and dilute 250 times, the medicine that above-mentioned dilution is good has every hole, hole of cell to add 50 μ L to kind, this moment, drug dilution was 1000 times, and promptly final concentration is 50 μ g/mL.Hatch 48h.Each concentration is established 4 (2) individual parallel multiple holes, and every plate is established negative control group (add blank solution in the cell, blank solution is joined method---add the culture fluid of 200 μ L, will draw 0.88 μ LDMSO adding mixing), positive controls (amycin final concentration 4 μ g/mL).
3SRB dyeing:
After cell culture finishes, take out culture plate, every hole adds trichloroacetic acid (TCA) the 70 μ L fixed cells of 40% (mass/volume), places 1h in 4 ℃ of refrigerators.Deionized water wash 5 times of each hole of culture plate are to remove TCA.Behind air drying, every hole adds 0.4% SRB100 μ L (dissolving of 1% chromatographically pure acetic acid), place 20min under the room temperature, discard in each hole and wash 5 times with 1% acetic acid behind the liquid, remove unconjugated dyestuff, with 10mmol/L Tris150 μ L/ hole dissolving, 5min vibrates behind the air drying, use microplate reader (ELx 800) to measure, used wavelength is 490nm.
The calculating suppression ratio of 4 suppression ratio is calculated as follows:
Drug effect the results are shown in Table 4.
Table 4, K562 inhibition rate of tumor cell result
| C06 | Negative | Blank | Positive | |
| Average cell survival number | 0.16 | 0.79 | 0.08 | 0.14 |
| Suppression ratio (%) | 88.42 | 0.00 | 100.00 | 91.20 |
| RSD(％) | 3.31 | 1.23 | 6.80 | 7.35 |
| C07 | Negative | Blank | Positive | |
| Average cell survival number | 0.167 | 0.725 | 0.062 | 0.106 |
| Suppression ratio (%) | 84.16 | 0 | 100 | 93.439 |
| RSD(％) | 6.801 | 2.355 | 4.878 | 6.118 |
Cell strain: KB tumor cell
Sample preparation: according to the weight of the medicine of institute's weighing, add the DMSO dissolving of respective volume, concentration is about 50mg/mL.The concussion dissolving, if there are a large amount of drops to glue wall, can be suitably centrifugal.Can store-20 ℃.
Cell culture: use DMEM high glycoform (Gibco) [adding the 3.7g/L sodium bicarbonate] 90%, calf serum (match is happy) 10%, non essential amino acid (Gibco) 1% Mixed culture KB cell.
Experimental technique:
1 kind of plate:
(1) calculates and to need cell total amount NT=ρ cellmL-1 * VT (ρ=2 * 103/mL), wherein VT=0.1mL * hole count+cell groove surplus.
(2) suspension cell on the piping and druming culture bottle wall adds in the centrifuge tube.Get 10 μ L, add the blue dilution of 10 μ L Placenta Hominiss, viable count sum NL on the count plate, then the cell number N in the centrifuge tube is: NL/4 * 104 * 2 * V, wherein V is the liquor capacity in the centrifugal preceding centrifuge tube.
It is (3) centrifugal that (900rad/min 10min), inhales and removes supernatant, adds culture fluid V1mL, and piping and druming makes the cell mixing, draws V2mL and joins in the cell groove, makes V2=NT/N * V1.
(4) add the culture fluid of VT-V2mL again in the cell groove, with volley of rifle fire piping and druming, mixing, get this liquid, every hole adds 100 μ L, hatches 24h.
(5) choose 4 holes behind the kind plate and add culture fluid as blank, the residue hole adds 100 μ L PBS, to reduce the evaporation of culture fluid.
2 dosings:
(1) 96 orifice plate changes liquid, and every hole adds fresh medium 150 μ L.
(2) culture fluid in adding 220 μ L/ holes in 96 new orifice plates, to draw 0.88 μ L medicinal liquid and add mixing, and dilute 250 times, the medicine that above-mentioned dilution is good has every hole, hole of cell to add 50 μ L to kind, this moment, drug dilution was 1000 times, and promptly final concentration is 50 μ g/mL.Hatch 48h.Each concentration is established 4 (2) individual parallel multiple holes, and every plate is established negative control group (add blank solution in the cell, blank solution is joined method---add the culture fluid of 200 μ L, will draw 0.88 μ LDMSO adding mixing), positive controls (amycin final concentration 4 μ g/mL).
The 3MTT colorimetric method for determining:
(1) take out culture plate, every hole, place to go supernatant, adding culture fluid-MTT mixed solution (culture fluid: 100 μ L MTT solution=10: 1), hatch 4h.
(2) culture fluid is abandoned in suction, and every hole adds the DMSO of 150 μ L, vibration 10min, and 550nm microplate reader (Elx800) is measured.
The calculating suppression ratio of 4 suppression ratio is calculated as follows:
Drug effect the results are shown in Table 5.
Table 5, KB inhibition rate of tumor cell result
| C06 | Negative | Blank | Positive | |
| Average cell survival number | 0.099 | 0.879 | 0.088 | 0.077 |
| Suppression ratio (%) | 98.609 | 0.000 | 100.000 | 101.391 |
| RSD(％) | 8.123 | 3.441 | 9.452 | 4.622 |
| C07 | Negative | Blank | Positive | |
| Average cell survival number | 0.157 | 0.879 | 0.088 | 0.077 |
| Suppression ratio (%) | 91.340 | 0.000 | 100.000 | 101.391 |
| RSD(％) | 5.072 | 3.441 | 9.452 | 4.622 |
Cell strain: MCF-7 tumor cell
Make up a prescription: according to the weight of the medicine of institute's weighing, add the DMSO dissolving of respective volume, concentration is about 50mg/mL.The concussion dissolving, if there are a large amount of drops to glue wall, can be suitably centrifugal.Can store-20 ℃.
Cell culture: use DMEM high glycoform (Gibco) [adding the 3.7g/L sodium bicarbonate] 90%, calf serum (match is happy) 10%, non essential amino acid (Gibco) 1% Mixed culture MCF-7 cell.
Experimental technique:
1 kind of plate:
(1) calculates and to need cell total amount NT=ρ cellmL-1 * VT (ρ=2 * 103/mL), wherein VT=0.1mL * hole count+cell groove surplus.
(2) suspension cell on the piping and druming culture bottle wall adds in the centrifuge tube.Get 10 μ L, add the blue dilution of 10 μ L Placenta Hominiss, viable count sum NL on the count plate, then the cell number N in the centrifuge tube is: NL/4 * 104 * 2 * V, wherein V is the liquor capacity in the centrifugal preceding centrifuge tube.
It is (3) centrifugal that (900rad/min 10min), inhales and removes supernatant, adds culture fluid V1mL, and piping and druming makes the cell mixing, draws V2mL and joins in the cell groove, makes V2=NT/N * V1.
(4) add the culture fluid of VT-V2mL again in the cell groove, with volley of rifle fire piping and druming, mixing, get this liquid, every hole adds 100 μ L, hatches 24h.
(5) choose 4 holes behind the kind plate and add culture fluid as blank, the residue hole adds 100 μ LPBS, to reduce the evaporation of culture fluid.
2 dosings:
(1) 96 orifice plate changes liquid, and every hole adds fresh medium 150 μ L.
(2) culture fluid in adding 220 μ L/ holes in 96 new orifice plates, to draw 0.88 μ L medicinal liquid and add mixing, and dilute 250 times, the medicine that above-mentioned dilution is good has every hole, hole of cell to add 50 μ L to kind, this moment, drug dilution was 1000 times, and promptly final concentration is 50 μ g/mL.Hatch 48h.Each concentration is established 4 (2) individual parallel multiple holes, and every plate is established negative control group (add blank solution in the cell, blank solution is joined method---add the culture fluid of 200 μ L, will draw 0.88 μ LDMSO adding mixing), positive controls (amycin final concentration 4 μ g/mL).
The 3MTT colorimetric method for determining:
(1) take out culture plate, every hole, place to go supernatant, adding culture fluid-MTT mixed solution (culture fluid: 100 μ L MTT solution=10: 1), hatch 4h.
(2) culture fluid is abandoned in suction, and every hole adds the DMSO of 150 μ L, vibration 10min, and 550nm microplate reader (Elx800) is measured.
The calculating suppression ratio of 4 suppression ratio is calculated as follows:
Drug effect the results are shown in Table 6.
Table 6, MCF-7 inhibition rate of tumor cell result
| Spica Prunellae C06 | Negative | Blank | Positive | |
| Average cell survival number | 0.226 | 0.417 | 0.065 | 0.105 |
| Suppression ratio (%) | 54.403 | 0.000 | 100.000 | 88.565 |
| RSD(％) | 14.016 | 4.888 | 4.031 | 3.415 |
| Spica Prunellae C07 | Negative | Blank | Positive | |
| Average cell survival number | 0.254 | 0.417 | 0.065 | 0.105 |
| Suppression ratio (%) | 46.449 | 0.000 | 100.000 | 88.565 |
| RSD(％) | 5.943 | 4.888 | 4.031 | 3.415 |
Pharmacological model: Hep G2 tumor cell
Cell culture and kind plateCell culture: use DMEM high glycoform (Gibco) [adding the 3.7g/L sodium bicarbonate] 90%, hyclone (Ilex purpurea Hassk.[I.chinensis Sims) 10%, non essential amino acid (Gibco) 1% Mixed culture Hep G2 cell.Calculating needs cell total amount NT=ρ cellmL-1 * VT (ρ=2 * 103/mL), wherein VT=0.1mL * hole count+cell groove surplus.Suspension cell on the piping and druming culture bottle wall adds in the centrifuge tube.Get 10 μ L, add the blue dilution of 10 μ L Placenta Hominiss, viable count sum NL on the count plate, then the cell number N in the centrifuge tube is: NL/4 * 104 * 2 * V, wherein V is the liquor capacity in the centrifugal preceding centrifuge tube.It is centrifugal that (900rad/min 10min), inhales and removes supernatant, adds culture fluid V1mL, and piping and druming makes the cell mixing, draws V2mL and joins in the cell groove, makes V2=NT/N * V1.The culture fluid that adds VT-V2mL in the cell groove again with volley of rifle fire piping and druming, mixing, is got this liquid, and every hole adds 100 μ L, hatches 24h.Choose 4 holes adding culture fluid as blank after planting plate, the residue hole adds 100 μ LPBS, to reduce the evaporation of culture fluid.
Dosage regimenSpica Prunellae C06 active component is according to the weight of the medicine of institute's weighing, according to the weight of the medicine of institute's weighing, adds the DMSO dissolving of respective volume, and concentration is about 50mg/mL.The concussion dissolving, if there are a large amount of drops to glue wall, can be suitably centrifugal.Can store-20 ℃.96 orifice plates change liquid, and every hole adds fresh medium 150 μ L.The culture fluid that adds 220 μ L/ holes in 96 new orifice plates will be drawn 0.88 μ L medicinal liquid and add mixing, dilute 250 times, and the medicine that above-mentioned dilution is good has every hole, hole of cell to add 50 μ L to kind, and this moment, drug dilution was 1000 times, and promptly final concentration is 50 μ g/mL.Hatch 48h.Each concentration is established 4 (2) individual parallel multiple holes, and every plate is established negative control group (add blank solution in the cell, blank solution is joined method---add the culture fluid of 200 μ L, will draw 0.88 μ LDMSO adding mixing), positive controls (amycin final concentration 4 μ g/mL).The MTT colorimetric method for determining: take out culture plate, every hole, place to go supernatant, adding culture fluid-MTT mixed solution (culture fluid: 100 μ L MTT solution=10: 1), hatch 4h.Culture fluid is abandoned in suction, and every hole adds the DMSO of 150 μ L, vibration 10min, and 550nm microplate reader (Elx800) is measured.
The calculating of suppression ratioSuppression ratio is calculated as follows:
Drug effect the results are shown in Table 7.
Table 7, Hep G2 inhibition rate of tumor cell result
| Spica Prunellae C06 component | Negative | Blank | Positive | |
| Average cell survival number | 0.093 | 0.256 | 0.069 | 0.082 |
| Suppression ratio (%) | 87.17 | 0 | 100 | 93.32 |
| RSD(％) | 6.208 | 4.297 | 2.734 | 8.762 |
| Spica Prunellae C07 component | Negative | Blank | Positive | |
| Average cell survival number | 0.191 | 0.665 | 0.164 | 0.196 |
| Suppression ratio (%) | 94.66 | 0 | 100 | 93.71 |
| RSD(％) | 5.107 | 0.956 | 3.884 | 4.986 |
Spica Prunellae active component of the present invention and preparation thereof can be used in preparation treatment, prophylaxis of tumours medicine.
Beneficial effect of the present invention is:
1. use normal phase silicagel column in the extraction and separation process of the present invention, can remove impurity such as desaccharide, protein, aminoacid effectively, improved content of effective, adopted preparative hplc simultaneously, can obtain effective ingredient fast and accurately.
2. Spica Prunellae active component chemical constituent provided by the invention is simply clear and definite, is easier to illustrate its mechanism of action on pharmacological research, is easier to the quality control of medicine aborning.
Method provided by the invention obtains containing C06 first from the Spica Prunellae medical material, the C07 active component, and first it is carried out medicine efficacy screening on various tumor cell strains, because composition is definite, content is clear and definite, and preparation technology is convenient, active good, the suitable antitumor new Chinese medicine that is developed to.
Description of drawings
Fig. 1 is the HPLC analysis chart of Spica Prunellae active component 1 of the present invention.
Fig. 2 is the HPLC analysis chart of Spica Prunellae active component 2 of the present invention.
The specific embodiment
Further describe flesh and blood of the present invention below in conjunction with embodiments of the invention, this embodiment only is used to the present invention is described and the present invention is not limited.
The preparation of embodiment 1 Spica Prunellae active component
Get Spica Prunellae medical material 250g, it is pulverized the back add ethyl acetate and ethanol (1: 1), reflux 1 hour is extracted 2 times, filtrate merge extracting solution.Extracting solution is condensed into extractum gets 16.6g, with extractum and silica gel mixed sample, separate with normal phase silicagel column, at first use petroleum ether and ethyl acetate (50: 1) as mobile phase, get eluent I, change chloroform and methanol (10: 1) then as mobile phase, get eluent II, get the 5.0g sample behind the concentrate drying.Continue to separate the sample that obtains with preparative liquid chromatography.The separation condition of preparative hplc: chromatographic column is Agient preparative column (Zorbax SB-C
1821.2mm * 250mm), mobile phase is water (A) and acetonitrile (B), the gradient elution program is as follows:
Flow velocity is 10ml/min, and column temperature is a room temperature.Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 24.0-28.0 minute or 28.0-32.0 minute, obtains two kinds of active component components 1 (0.36g) and component 2 (0.17g) behind the concentrate drying, each.
The HPLC-ELSD of embodiment 2 Spica Prunellae active components analyzes
Chromatographic conditionChromatographic column Agilent Zorbax SB-C18 post (4.6mm * 150mm, 5 μ m); Adopt gradient elution, mobile phase A is 0.2% glacial acetic acid aqueous solution mutually, and Mobile phase B is mutually for containing the acetonitrile solution of 0.2% glacial acetic acid; The gradient elution program is as follows: in the time of 0 minute, mobile phase A is that 90% 0.2% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 10% 0.2% glacial acetic acid; In the time of 10 minutes, mobile phase A is that 50% 0.2% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 50% 0.2% glacial acetic acid; In the time of 30 minutes, mobile phase A is that 5% 0.2% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile solution of 95% 0.2% glacial acetic acid; In the time of 35 minutes, mobile phase A is that 5% 0.2% glacial acetic acid aqueous solution, Mobile phase B are the acetonitrile of 95% 0.2% glacial acetic acid.Solution flow rate 0.5mLmin
-1It is long to detect the wavelength all-wave; 30 ℃ of column temperatures; ELSD condition: 105 ℃ of drift tube temperatures; Nitrogen flow rate 2.0L/min
The preparation of need testing solutionTake by weighing active component of the present invention, in volumetric flask, be diluted to scale, shake up, promptly with dissolve with methanol solution.
Assay methodThe accurate need testing solution of drawing injects chromatograph of liquid, measures.
Embodiment 3 Spica Prunellae active component preparations
Get Spica Prunellae active component 1 or the component 2 of embodiment 1,0.5g and 10.5g Polyethylene Glycol-6000 mix homogeneously, heating and melting moves in the drop pill drip irrigation behind the change material, and medicine liquid droplet is to 6-8 ℃ of liquid paraffin, and oil removing makes 400 of drop pill.
Get Spica Prunellae active component 1 or the component 2 of embodiment 1,0.5g, glucose 4.5g, sodium thiosulfate 0.9g and distilled water 1ml, behind the said components mix homogeneously, lyophilization, 500 of packing, promptly.
Get Lignum Dalbergiae Odoriferae oil 1.5g, join in the saturated hydroxypropyl of 13ml, stirring and dissolving filters, the filter leaf cold drying, Lignum Dalbergiae Odoriferae oil and the clathrate powder of hydroxypropyl.Except that above-mentioned Lignum Dalbergiae Odoriferae oil closes the clathrate powder of hydroxypropyl, get Spica Prunellae active component 1 or the component 2 of embodiment 1 again, 0.5g, mannitol 5.5g, calcium disodium edetate 0.9g and distilled water 2ml, behind the said components mixing, lyophilization, 300 of packing, promptly.
The preparation of embodiment 6 Spica Prunellae active components
Operating procedure is with embodiment 1, and condition changes into: ethyl acetate and ethanol (1: 5), normal phase silicagel column separate, and at first use petroleum ether and ethyl acetate 47: 1 as mobile phase, eluent I, change chloroform and methanol (8: 1) then as mobile phase.
The preparation of embodiment 7 Spica Prunellae active components
Operating procedure is with embodiment 1, and condition changes into: ethyl acetate and ethanol (5: 1), normal phase silicagel column separate, and at first use petroleum ether and ethyl acetate 53: 1 as mobile phase, eluent I, change chloroform and methanol (13: 1) then as mobile phase.
The preparation of embodiment 8 Spica Prunellae active components
Operating procedure is with embodiment 1, and condition changes into: ethyl acetate and ethanol (1: 2), normal phase silicagel column separate, and at first use petroleum ether and ethyl acetate 47: 1 as mobile phase, eluent I, change chloroform and methanol (8: 1) then as mobile phase.
The preparation of embodiment 9 Spica Prunellae active components
Operating procedure is with embodiment 1, and condition changes into: ethyl acetate and ethanol (2: 1), normal phase silicagel column separate, and at first use petroleum ether and ethyl acetate 53: 1 as mobile phase, eluent I, change chloroform and methanol (13: 1) then as mobile phase.
Claims (10)
1, a kind of Spica Prunellae active component is characterized in that, its preparation process may further comprise the steps:
Step 1: Spica Prunellae is extracted as solvent with ethyl acetate and alcohol mixture,
Step 2: extracting solution gets eluent through column chromatography;
Step 3: with the eluent that the preparative liquid chromatography gradient elution obtains, mobile phase is water and acetonitrile, collects 24.0-28.0 minute or 28.0-32.0 minute eluent obtains active component.
2, the active component of claim 1 is characterized in that, ethyl acetate described in the step 1 and alcoholic acid mixture, and both ratios are ethyl acetate: ethanol=1-5: 1-5.
3, the active component of claim 1 is characterized in that, ethyl acetate described in the step 1 and alcoholic acid mixture, and both ratios are ethyl acetate: ethanol=1-2: 1-2.
4, the active component of claim 1 is characterized in that, ethyl acetate described in the step 1 and alcoholic acid mixture, and both ratios are ethyl acetate: ethanol=1: 1.
5, the active component of claim 1, it is characterized in that, in the described step, step 1 is: get the Spica Prunellae medical material, with ethyl acetate: ethanol=1-5: 1-5 is solvent, reflux, extract,, extracting solution is separated with medicinal residues, and the extracting solution that obtains after the separation is an extract 1, and step 2 is: extract 1 is crossed normal phase silicagel column, earlier use petroleum ether: ethyl acetate=47～53: 1 as the mobile phase eluting, change chloroform then: methanol 8～13: 1 as mobile phase, gets eluent, and step 3 is: continue to separate the eluent that obtains with preparative liquid chromatography, mobile phase is water-A and acetonitrile-B, carries out gradient elution.
6, the active component of claim 5 is characterized in that, described gradient elution program is as follows:
Flow velocity is 10ml/min, and column temperature is a room temperature; Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 24.0-28.0 minute or 28.0-32.0 minute, and solution obtains active component behind concentrate drying.
7, the pharmaceutical composition that contains any one active component of claim 1-6.
8, the preparation method of the active component of claim 1 is characterized in that, its preparation process may further comprise the steps:
Step 1: Spica Prunellae is extracted as solvent with ethyl acetate and alcohol mixture,
Step 2: extracting solution gets eluent through column chromatography;
Step 3: with the eluent that the preparative liquid chromatography gradient elution obtains, mobile phase is water and acetonitrile, collects 24.0-28.0 minute or 28.0-32.0 minute eluent obtains active component.
9, the preparation method of the active component of claim 8, it is characterized in that, in the described step, step 1 is: get the Spica Prunellae medical material, with ethyl acetate: ethanol=1-5: 1-5 is solvent, reflux, extract,, extracting solution is separated with medicinal residues, and the extracting solution that obtains after the separation is an extract 1, and step 2 is: extract 1 is crossed normal phase silicagel column, earlier use petroleum ether: ethyl acetate=47～53: 1 as the mobile phase eluting, change chloroform then: methanol 8～13: 1 as mobile phase, gets eluent, and step 3 is: continue to separate the eluent that obtains with preparative liquid chromatography, mobile phase is water-A and acetonitrile-B, and the gradient elution program is as follows:
Flow velocity is 10ml/min, and column temperature is a room temperature; Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 24.0-28.0 minute or 28.0-32.0 minute, and solution obtains active component behind concentrate drying.
10, the preparation method of the active component of claim 9 is characterized in that, step is as follows:
To add ethyl acetate after the Spica Prunellae pulverizing medicinal materials: ethanol=1: 1, reflux 1 hour is extracted 2 times, and merging filtrate gets extracting solution; Extracting solution is condensed into extractum, and itself and silica gel mixed sample, with normal phase silicagel column it is separated, at first use petroleum ether: ethyl acetate=50: 1 is as mobile phase, eluent I, abandon it, change chloroform then: methanol=10: 1 is as mobile phase, get eluent II, will get sample behind the eluent II concentrate drying; Continue to separate the sample that obtains with preparative liquid chromatography; The separation condition of preparative hplc: chromatographic column is preparative column Zorbax SB-C18; 21.2mm * 250mm, mobile phase is water A and acetonitrile B, and the gradient elution program is as follows:
Flow velocity is 10ml/min, and column temperature is a room temperature; Sample 100% dissolve with ethanol separates through preparative liquid chromatography, collects solution at time period 24.0-28.0 minute or 28.0-32.0 minute, and solution obtains active component behind concentrate drying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007101232735A CN101347502B (en) | 2007-07-20 | 2007-07-20 | Effective component of selfheal and preparation and use thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007101232735A CN101347502B (en) | 2007-07-20 | 2007-07-20 | Effective component of selfheal and preparation and use thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101347502A true CN101347502A (en) | 2009-01-21 |
| CN101347502B CN101347502B (en) | 2012-01-04 |
Family
ID=40266583
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2007101232735A Expired - Fee Related CN101347502B (en) | 2007-07-20 | 2007-07-20 | Effective component of selfheal and preparation and use thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101347502B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673495A (en) * | 2015-02-10 | 2015-06-03 | 湖南中医药大学 | Method for extracting volatile oil for preventing and treating gynecologic inflammation from self-heal |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1052412C (en) * | 1994-08-09 | 2000-05-17 | 马洪万 | Cancer therapy injection |
-
2007
- 2007-07-20 CN CN2007101232735A patent/CN101347502B/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673495A (en) * | 2015-02-10 | 2015-06-03 | 湖南中医药大学 | Method for extracting volatile oil for preventing and treating gynecologic inflammation from self-heal |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101347502B (en) | 2012-01-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101347501B (en) | Effective component of Rabdosia amethystoides and preparation and use thereof | |
| CN101347518B (en) | Effective component of red paeonia and preparation and use thereof | |
| CN101347497B (en) | Effective component of vietnamese sophora root and preparation and use thereof | |
| CN101347521B (en) | Effective component of Clematis chinensis and preparation and use thereof | |
| CN101347484A (en) | Effective component of Atractylodes macrocephala and preparation and use thereof | |
| CN101347496B (en) | Effective component of Spatholobus stem and preparation and use thereof | |
| CN101347500B (en) | Effective component of rabdosia and preparation and use thereof | |
| CN101347473B (en) | Effective component of creat and preparation and use thereof | |
| CN101549111B (en) | Effective component of cogongrass rhizome and preparation method thereof | |
| CN101549000B (en) | Effective component of cynanchum atratum and preparation method thereof | |
| CN101347509B (en) | Effective component of radix stephaniae tetrandrae and preparation method thereof | |
| CN101347502B (en) | Effective component of selfheal and preparation and use thereof | |
| CN101347480B (en) | Effective component of inula flower and preparation and use thereof | |
| CN101549048B (en) | Effective component of vervain and preparation method thereof | |
| CN101347597B (en) | Effective component of turmeric rhizome and preparation method and use thereof | |
| CN101347467B (en) | Effective component of common camptotheca fruit and preparation as well as use thereof | |
| CN101347508B (en) | Effective component of Sinomenium acutum and preparation thereof | |
| CN101428120B (en) | Effective component of phragmites communis trin, preparation method and use thereof | |
| CN101428071B (en) | Effective component of pulsatilla, preparation method and use thereof | |
| CN101433614B (en) | Effective component of Chinese dittany bark as well as preparation method and use thereof | |
| CN101428053B (en) | Effective component of cowherb seed and preparation method and application thereof | |
| CN101347566B (en) | Effective component of airpotato yam as well as preparation method and use thereof | |
| CN101347468B (en) | Effective component of common camptotheca fruit and preparation as well as use thereof | |
| CN101428129B (en) | Effective component of turmeric rhizome, preparation method and uses thereof | |
| CN101433570B (en) | Effective component of cowherb seed as well as preparation method and use thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee |
Owner name: TASLY PHARMACEUTICAL GROUP CO., LTD. Free format text: FORMER NAME: TIANJIN TIANSHILI PHARMACEUTICAL CO., LTD. |
|
| CP01 | Change in the name or title of a patent holder |
Address after: 300402 Tianjin Beichen Xinyi white road Liaohe Road No. 1 Patentee after: TASLY PHARMACEUTICAL GROUP Co.,Ltd. Address before: 300402 Tianjin Beichen Xinyi white road Liaohe Road No. 1 Patentee before: TIANJIN TASLY PHARMACEUTICAL Co.,Ltd. |
|
| CP03 | Change of name, title or address | ||
| CP03 | Change of name, title or address |
Address after: 300410 Tianjin city Beichen District Huaihe road and road intersection Dingjiang tianzhijiao Park forensic Center for Intellectual Property Department Patentee after: TASLY PHARMACEUTICAL GROUP Co.,Ltd. Address before: 300402 Tianjin Beichen Xinyi white road Liaohe Road No. 1 Patentee before: TASLY PHARMACEUTICAL GROUP Co.,Ltd. |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20120104 Termination date: 20210720 |