CN101978962B - Medicinal composition for colon cancer and method for preparing effective monomers thereof - Google Patents
Medicinal composition for colon cancer and method for preparing effective monomers thereof Download PDFInfo
- Publication number
- CN101978962B CN101978962B CN2010102868557A CN201010286855A CN101978962B CN 101978962 B CN101978962 B CN 101978962B CN 2010102868557 A CN2010102868557 A CN 2010102868557A CN 201010286855 A CN201010286855 A CN 201010286855A CN 101978962 B CN101978962 B CN 101978962B
- Authority
- CN
- China
- Prior art keywords
- berberine
- rutaecarpine
- rutaecarpin
- value
- colon cancer
- 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.)
- Active
Links
- 206010009944 Colon cancer Diseases 0.000 title claims abstract description 30
- 208000029742 colonic neoplasm Diseases 0.000 title claims abstract description 26
- 239000000203 mixture Substances 0.000 title claims abstract description 13
- 238000000034 method Methods 0.000 title abstract description 47
- 239000000178 monomer Substances 0.000 title abstract description 12
- ACVGWSKVRYFWRP-UHFFFAOYSA-N Rutecarpine Chemical compound C1=CC=C2C(=O)N(CCC=3C4=CC=CC=C4NC=33)C3=NC2=C1 ACVGWSKVRYFWRP-UHFFFAOYSA-N 0.000 claims abstract description 213
- YBHILYKTIRIUTE-UHFFFAOYSA-N berberine Chemical compound C1=C2CC[N+]3=CC4=C(OC)C(OC)=CC=C4C=C3C2=CC2=C1OCO2 YBHILYKTIRIUTE-UHFFFAOYSA-N 0.000 claims abstract description 88
- 229940093265 berberine Drugs 0.000 claims abstract description 88
- QISXPYZVZJBNDM-UHFFFAOYSA-N berberine Natural products COc1ccc2C=C3N(Cc2c1OC)C=Cc4cc5OCOc5cc34 QISXPYZVZJBNDM-UHFFFAOYSA-N 0.000 claims abstract description 88
- BCPAKGGXGLGKIO-UHFFFAOYSA-N Pseudorutaecarpin Natural products C1=CC=C2C(=O)N(CCC3=C4C5=CC=CC=C5N3)C4=NC2=C1 BCPAKGGXGLGKIO-UHFFFAOYSA-N 0.000 claims abstract description 70
- 238000002360 preparation method Methods 0.000 claims description 26
- 239000003814 drug Substances 0.000 claims description 19
- 239000008194 pharmaceutical composition Substances 0.000 claims description 11
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- 239000008187 granular material Substances 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 239000002552 dosage form Substances 0.000 claims 1
- 239000003826 tablet Substances 0.000 claims 1
- 241000700159 Rattus Species 0.000 abstract description 44
- 201000011510 cancer Diseases 0.000 abstract description 19
- 206010028980 Neoplasm Diseases 0.000 abstract description 15
- 210000002700 urine Anatomy 0.000 abstract description 14
- DIIIISSCIXVANO-UHFFFAOYSA-N 1,2-Dimethylhydrazine Chemical compound CNNC DIIIISSCIXVANO-UHFFFAOYSA-N 0.000 abstract description 11
- 201000002313 intestinal cancer Diseases 0.000 abstract description 8
- 230000002503 metabolic effect Effects 0.000 abstract description 8
- 208000005016 Intestinal Neoplasms Diseases 0.000 abstract description 7
- 238000005516 engineering process Methods 0.000 abstract description 2
- 238000012216 screening Methods 0.000 abstract description 2
- TXDUTHBFYKGSAH-SFHVURJKSA-N Evodiamine Chemical compound C1=CC=C2N(C)[C@@H]3C(NC=4C5=CC=CC=4)=C5CCN3C(=O)C2=C1 TXDUTHBFYKGSAH-SFHVURJKSA-N 0.000 abstract 3
- HMXRXBIGGYUEAX-SFHVURJKSA-N Evodiamine Natural products CN1[C@H]2N(CCc3[nH]c4ccccc4c23)C(=O)c5ccccc15 HMXRXBIGGYUEAX-SFHVURJKSA-N 0.000 abstract 3
- 241000209020 Cornus Species 0.000 abstract 1
- 238000001819 mass spectrum Methods 0.000 abstract 1
- 238000005259 measurement Methods 0.000 abstract 1
- 230000036285 pathological change Effects 0.000 abstract 1
- 231100000915 pathological change Toxicity 0.000 abstract 1
- 239000006187 pill Substances 0.000 abstract 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 114
- 235000019441 ethanol Nutrition 0.000 description 57
- 238000000151 deposition Methods 0.000 description 52
- 230000008021 deposition Effects 0.000 description 52
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 48
- 239000000243 solution Substances 0.000 description 46
- 239000013078 crystal Substances 0.000 description 43
- 238000000605 extraction Methods 0.000 description 39
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 38
- 238000001953 recrystallisation Methods 0.000 description 38
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 36
- 229960004756 ethanol Drugs 0.000 description 35
- 238000001914 filtration Methods 0.000 description 29
- 239000000843 powder Substances 0.000 description 27
- 229910021529 ammonia Inorganic materials 0.000 description 24
- 230000001105 regulatory effect Effects 0.000 description 24
- 230000008569 process Effects 0.000 description 22
- 238000002481 ethanol extraction Methods 0.000 description 21
- 239000000284 extract Substances 0.000 description 21
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- 229960000583 acetic acid Drugs 0.000 description 18
- 239000012362 glacial acetic acid Substances 0.000 description 18
- 239000003960 organic solvent Substances 0.000 description 18
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 17
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 17
- 239000000920 calcium hydroxide Substances 0.000 description 17
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 17
- 238000004062 sedimentation Methods 0.000 description 17
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 16
- 239000000706 filtrate Substances 0.000 description 16
- 239000003513 alkali Substances 0.000 description 15
- 239000012141 concentrate Substances 0.000 description 15
- 230000000694 effects Effects 0.000 description 15
- 230000004060 metabolic process Effects 0.000 description 15
- 230000003902 lesion Effects 0.000 description 14
- 238000001556 precipitation Methods 0.000 description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 14
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 12
- 208000035126 Facies Diseases 0.000 description 12
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 12
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 12
- OAIVIYSBZFEOIU-UHFFFAOYSA-N chloroform;propan-2-one Chemical compound CC(C)=O.ClC(Cl)Cl OAIVIYSBZFEOIU-UHFFFAOYSA-N 0.000 description 12
- 239000000463 material Substances 0.000 description 12
- 238000010992 reflux Methods 0.000 description 12
- 239000001117 sulphuric acid Substances 0.000 description 12
- 235000011149 sulphuric acid Nutrition 0.000 description 12
- 238000011282 treatment Methods 0.000 description 12
- 230000002378 acidificating effect Effects 0.000 description 11
- 230000001476 alcoholic effect Effects 0.000 description 11
- 238000010298 pulverizing process Methods 0.000 description 11
- 238000002390 rotary evaporation Methods 0.000 description 11
- 238000005406 washing Methods 0.000 description 11
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 150000001875 compounds Chemical group 0.000 description 9
- 238000001212 derivatisation Methods 0.000 description 9
- 238000011160 research Methods 0.000 description 9
- 239000002904 solvent Substances 0.000 description 9
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 8
- 229940079593 drug Drugs 0.000 description 8
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 8
- 230000036541 health Effects 0.000 description 8
- 230000001373 regressive effect Effects 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- 230000008859 change Effects 0.000 description 6
- RIFGWPKJUGCATF-UHFFFAOYSA-N ethyl chloroformate Chemical compound CCOC(Cl)=O RIFGWPKJUGCATF-UHFFFAOYSA-N 0.000 description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 6
- 238000003909 pattern recognition Methods 0.000 description 6
- 230000037396 body weight Effects 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 238000002560 therapeutic procedure Methods 0.000 description 5
- 230000003442 weekly effect Effects 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 208000001333 Colorectal Neoplasms Diseases 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 238000010171 animal model Methods 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 239000012535 impurity Substances 0.000 description 4
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical group [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 4
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 4
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 239000002671 adjuvant Substances 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000013459 approach Methods 0.000 description 3
- 238000002512 chemotherapy Methods 0.000 description 3
- 238000004090 dissolution Methods 0.000 description 3
- 238000009837 dry grinding Methods 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000010606 normalization Methods 0.000 description 3
- 206010065553 Bone marrow failure Diseases 0.000 description 2
- 208000037051 Chromosomal Instability Diseases 0.000 description 2
- 241000218202 Coptis Species 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-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
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 2
- 108700020796 Oncogene Proteins 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 2
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 2
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 2
- -1 and 1 time weekly Chemical compound 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000007405 data analysis Methods 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 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 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 150000002170 ethers Chemical class 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 229960002949 fluorouracil Drugs 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 235000019359 magnesium stearate Nutrition 0.000 description 2
- 206010025482 malaise Diseases 0.000 description 2
- 239000012567 medical material Substances 0.000 description 2
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 2
- 239000008108 microcrystalline cellulose Substances 0.000 description 2
- 229940016286 microcrystalline cellulose Drugs 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- 238000002203 pretreatment Methods 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 210000002784 stomach Anatomy 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 238000010792 warming Methods 0.000 description 2
- CVZZNRXMDCOHBG-QMMMGPOBSA-N 2-Chloro-L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1Cl CVZZNRXMDCOHBG-QMMMGPOBSA-N 0.000 description 1
- 208000003200 Adenoma Diseases 0.000 description 1
- 206010001233 Adenoma benign Diseases 0.000 description 1
- 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 1
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 235000002991 Coptis groenlandica Nutrition 0.000 description 1
- XDVZNDLANFJOQR-UHFFFAOYSA-N Coptisine Natural products O=Cc1c2OCOc2ccc1C=C3/NCCc4cc5OCOc5cc34 XDVZNDLANFJOQR-UHFFFAOYSA-N 0.000 description 1
- CMSMOCZEIVJLDB-UHFFFAOYSA-N Cyclophosphamide Chemical compound ClCCN(CCCl)P1(=O)NCCCO1 CMSMOCZEIVJLDB-UHFFFAOYSA-N 0.000 description 1
- 201000003741 Gastrointestinal carcinoma Diseases 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- MXTLAHSTUOXGQF-UHFFFAOYSA-O Jatrorrhizine Chemical compound COC1=CC=C2C=C3C(C=C(C(=C4)O)OC)=C4CC[N+]3=CC2=C1OC MXTLAHSTUOXGQF-UHFFFAOYSA-O 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 229930192392 Mitomycin Natural products 0.000 description 1
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 1
- PTPHDVKWAYIFRX-UHFFFAOYSA-N Palmatine Natural products C1C2=C(OC)C(OC)=CC=C2C=C2N1CCC1=C2C=C(OC)C(OC)=C1 PTPHDVKWAYIFRX-UHFFFAOYSA-N 0.000 description 1
- 241001250596 Pleione Species 0.000 description 1
- 208000015634 Rectal Neoplasms Diseases 0.000 description 1
- 241001093501 Rutaceae Species 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 208000007271 Substance Withdrawal Syndrome Diseases 0.000 description 1
- 241001078983 Tetradium ruticarpum Species 0.000 description 1
- 108010074506 Transfer Factor Proteins 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 206010047700 Vomiting Diseases 0.000 description 1
- 229960001138 acetylsalicylic acid Drugs 0.000 description 1
- 208000009956 adenocarcinoma Diseases 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000000118 anti-neoplastic effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 150000003836 berberines Chemical group 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 201000008275 breast carcinoma Diseases 0.000 description 1
- 229960000590 celecoxib Drugs 0.000 description 1
- RZEKVGVHFLEQIL-UHFFFAOYSA-N celecoxib Chemical compound C1=CC(C)=CC=C1C1=CC(C(F)(F)F)=NN1C1=CC=C(S(N)(=O)=O)C=C1 RZEKVGVHFLEQIL-UHFFFAOYSA-N 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- XYHOBCMEDLZUMP-UHFFFAOYSA-N coptisine Chemical compound C1=C2C=C(C3=C(C=C4OCOC4=C3)CC3)[N+]3=CC2=C2OCOC2=C1 XYHOBCMEDLZUMP-UHFFFAOYSA-N 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 229940111134 coxibs Drugs 0.000 description 1
- 239000003255 cyclooxygenase 2 inhibitor Substances 0.000 description 1
- 229960004397 cyclophosphamide Drugs 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000003748 differential diagnosis Methods 0.000 description 1
- 208000010643 digestive system disease Diseases 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 238000007908 dry granulation Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 239000003172 expectorant agent Substances 0.000 description 1
- 230000003419 expectorant effect Effects 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 230000030136 gastric emptying Effects 0.000 description 1
- 208000018685 gastrointestinal system disease Diseases 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 230000003760 hair shine Effects 0.000 description 1
- 235000008216 herbs Nutrition 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical class OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 1
- 238000003859 hyphenated technique Methods 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 230000001976 improved effect Effects 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000314 lubricant Chemical class 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000001035 methylating effect Effects 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 230000033607 mismatch repair Effects 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000002969 morbid Effects 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 229940126701 oral medication Drugs 0.000 description 1
- QUCQEUCGKKTEBI-UHFFFAOYSA-N palmatine Chemical compound COC1=CC=C2C=C(C3=C(C=C(C(=C3)OC)OC)CC3)[N+]3=CC2=C1OC QUCQEUCGKKTEBI-UHFFFAOYSA-N 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000011248 postoperative chemotherapy Methods 0.000 description 1
- 238000009702 powder compression Methods 0.000 description 1
- 238000000513 principal component analysis Methods 0.000 description 1
- 230000000750 progressive effect Effects 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 206010038038 rectal cancer Diseases 0.000 description 1
- 201000001275 rectum cancer Diseases 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000005070 ripening Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000008247 solid mixture Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 230000008093 supporting effect Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 102000003390 tumor necrosis factor Human genes 0.000 description 1
- 229940124549 vasodilator Drugs 0.000 description 1
- 239000003071 vasodilator agent Substances 0.000 description 1
Images
Abstract
The invention relates to a medicinal composition for colon cancer and a method for preparing effective monomers thereof. Based on the clinical classical golden cornus pill formula for treating the colon cancer, the medicinal composition consisting of berberine, evodiamine and rutaecarpine is obtained by compatibility and screening of multiple formulas. Urine of rats at different time points of a dimethylhydrazine-induced intestinal cancer model is measured by adopting gas chromatogram and mass spectrum combined technology, and the measurement result is combined with a mode identification method to discover that the metabolic track of the rats obviously regresses to that of healthy rats when the rats of a dimethylhydrazine-induced cancerous pathological change group and a cancer group take the medicinal composition consisting of the berberine, the evodiamine and the rutaecarpine in a proper ratio; and the composition consisting of the berberine, the evodiamine and the rutaecarpine can play a certain role in preventing and treating the colon cancer by controlling a body network.
Description
Technical field
The present invention relates to a kind of pharmaceutical composition that is used to prevent and/or treat colon cancer, wherein said compositions active component is made up of berberine, rutaecarpin and rutaecarpine.
Background technology
Intestinal cancer is the malignant tumor that big epithelium of intestinal mucosa and body of gland take place, and comprises colon cancer and rectal cancer.It is a kind of common malignancy, and interior world's most countries of more than 20 year time in the past or the morbidity of regional colorectal cancer are in rising trend, and rises more obvious with the low developing country of sickness rate.Intestinal cancer is western countries crowd's one of the reason of mainly dying of illness, and simultaneously, also is one of modal malignant tumor in the population of China, and its sickness rate is only second to pulmonary carcinoma and breast carcinoma in various malignant tumor.For many years, 5 years survival rates of colorectal cancer still rest on about 50% not because of progressive being significantly improved of performing the operation.
The Drug therapy of intestinal cancer; Mainly adopt three kinds of Therapeutic Method at present in the world: 1. chemotherapy; Postoperative chemotherapy of patients can use 2 to 3 courses of treatment in general 1 year to a year and a half, common drug mainly is 5-fluorouracil (5-FU), but also Combined application mitomycin, cyclophosphamide etc.Drug withdrawal in time when bone marrow depression is obvious.The oral medication gastrointestinal reaction is bigger than intravenously administrable, but the bone marrow depression response light.Because this medicine has obvious toxic and side effects, therefore, must be noted that Supporting Therapy during the medication, and Combined application reduces the medicine of side effect.2. immunization therapy; Can improve patient's antineoplastic ability, development in recent years is very fast, such as interferon, interleukin, transfer factor, tumor necrosis factor etc.; Extensive use gradually not only can improve patient's immunocompetence but also carrying out that can combined with chemotherapy.3. treatment by Chinese herbs can improve symptom, and the resistance against diseases of enhancing body reduces the side effect of radiotherapy, chemotherapy, and the Chinese medicine that has has direct antitumaous effect, like Herba Hedyotidis Diffusae, Herba Scutellariae Barbatae, Pseudobulbus cremastrae seu pleiones, Herba Solani Nigri etc.The superiority of Chinese medicine malignant tumor is admitted by numerous doctors and patients, and therapy of combining Chinese and Western medicine has special advantages at aspects such as improving life quality, blocking-up precancerous lesion.Can be dialectical during medication, differential diagnosis of diseases takes into account, add heat-clearing and toxic substances removing, invigorate blood circulation assault fortified position, the medicine of aspects such as nourishing YIN and benefiting blood, expectorant eliminating stagnation, harmonizing the spleen and stomach.Therefore, Chinese medicine has the incomparable advantage of Western medicine aspect the treatment intestinal cancer.
Zuojin Wan is the name side that comes from " danxi's experiential therapy fire six ", in that " Chinese pharmacopoeia is recorded, and is mainly used in the treatment gastroenteropathy clinically.Its prescription medical material Rhizoma Coptidis Coptis ch inensis Franch. is a Ranunculaceae Coptis herbaceos perennial; Effects such as tool heat clearing and damp drying, eliminating fire and detoxication; And good antibiotic, antiinflammatory action are arranged, main active is alkaloids compositions such as berberine, coptisine, palmatine and jateorhizine.Another prescription medical material Fructus Evodiae Evodia rutaeca rpa (Juss.) Benth is the fruit of the nearly drying and ripening of rutaceae Fructus Evodiae; Have effects such as stomach warming emesis, warming spleen and stopping diarrha, analgesia; Main active is chemical compounds such as rutaecarpin, rutaecarpine and polysaccharide, but vasodilator, promotes gastric emptying etc.Mainly concentrate on behind pharmacological action and the prescription different proportion compatibility of compound recipe aspects such as influence to the main component dissolution rate about the research of Zuojin Wan.Zuojin Wan and add the flavor Zuojin Wan and be used to the treatment of colon cancer at present clinically obtained certain clinical effectiveness, but real material base and mechanism of action of its treatment colon cancer is unclear.
The present invention is intended to the monomeric compound in the Zuojin Wan is carried out separation and purification obtaining monomer component, and carries out effective compatibility through above-mentioned monomer, filters out the best proportioning between them, with the pharmaceutical composition of the treatment colon cancer that obtains determined curative effect.
Summary of the invention
The invention reside in provides a kind of pharmaceutical composition of treating colon cancer; Said active ingredient in pharmaceutical is made up of berberine, rutaecarpin and rutaecarpine; Wherein said berberine: rutaecarpin: the ratio between the rutaecarpine is 5～10: 3～8: 1～5; Be preferably 5～7: 3～5: 1～2, most preferably be 6: 3: 1.
The present invention also provides separating of berberine, rutaecarpin and rutaecarpine and purification process; The method of the complicated drug effect effect of a kind of new evaluation is provided simultaneously, is used to instruct the rational use of drug that clinically Zuojin Wan is used for intestinal cancer.
The method for preparing of berberine is in a specific embodiments of the present invention: get fresh Rhizoma Coptidis and shine dry grinding, obtain the Rhizoma Coptidis coarse powder, coarse powder is used ethanol extraction, filters; Merging filtrate, filtrating concentrates, and puts coldly, adds glacial acetic acid; Separate out deposition, sedimentation and filtration is removed.Solution adds concentrated hydrochloric acid and regulates pH value, and placement is spent the night, and obtains yellow acicular crystal; Yellow acicular crystal is used the organic solvent recrystallization, regulates pH value with calcium hydroxide; Obtaining purity is more than 98%, and single related substance is all less than 0.1% berberine monomeric compound.Wherein, said glacial acetic acid is preferably 1% glacial acetic acid; Said adding concentrated hydrochloric acid is regulated in the step of pH value, and preferably regulating pH is 3-4; Said adding calcium hydroxide is regulated in the step of pH value, and preferably regulating pH is 7.5-10, and 7.5-8.0 more preferably, or 8.5-9.0 or 9.5-10 most preferably are 8.5-9.0.
In a preferred specific embodiments of the present invention, be preferably the ethanol extraction that adopts 5-15 doubly to measure in the method for preparing of berberine, 5 times of amount ethanol more preferably, or 10 times of amount ethanol, or 15 times of amount ethanol most preferably are 5 times of amount ethanol.
In a preferred specific embodiments of the present invention, extract the ethanol that uses in the method for preparing of berberine and be 10-95% ethanol, be preferably 30-75% 50-70% ethanol more preferably, most preferably be 60% ethanol.
In a preferred specific embodiments of the present invention, extraction time is 1-5 hour in the method for preparing of berberine, is preferably 1-4 hour, more preferably 2-3 hour, most preferably is 2 hours.
In a preferred specific embodiments of the present invention, extraction time is 1-5 time in the method for preparing of berberine, is preferably 1-4 time, more preferably 2-3 time, most preferably is 3 times.
In a preferred specific embodiments of the present invention, in the method for preparing of berberine in the step of recrystallization preferred organic be lower alcohol such as methanol, ethanol, propanol etc.; Chlorohydrocarbon such as chloroform, dichloromethane, carbon tetrachloride etc., ethers such as petroleum ether, ether etc., acetone; Ethyl acetate, oxolane, hexane etc., more preferably position lower alcohol such as methanol; Ethanol, propanol etc. most preferably are ethanol.
In more preferred specific embodiments of the present invention, the method for preparing of berberine is meant that specifically getting fresh Rhizoma Coptidis shines dry grinding, obtains the Rhizoma Coptidis coarse powder, and coarse powder is measured 60% ethanol extraction 3 times with 5 times; Each 2 hours, filter merging filtrate; Filtrating is concentrated into 40 ℃, and to measure relative densities be 1.45, puts coldly, adds 1% glacial acetic acid; Separate out a large amount of depositions, sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used ethyl alcohol recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, obtains purity and is approximately 98% berberine monomeric compound.
In another specific embodiments of the present invention, the method for preparing of rutaecarpin and rutaecarpine is: with using organic solvent extraction after the Fructus Evodiae pulverizing medicinal materials, said organic solvent is lower alcohol such as methanol, ethanol; Propanol etc., chlorohydrocarbon such as chloroform, dichloromethane, carbon tetrachloride etc., ethers such as petroleum ether, ether etc., acetone; Ethyl acetate, oxolane, hexane etc., more preferably position lower alcohol such as methanol; Ethanol, propanol etc. most preferably are ethanol; The extracting solution rotary evaporation is separated out to a small amount of deposition, and concentrate is preferably mineral acid and regulates pH value like 5% sulphuric acid with acid, and said pH value is preferably and is adjusted to 2-2.5; Place afterwards, obtain the brown insoluble matter, filter, filtrating is the dilute acidic alcoholic solution; Use alkali then, preferably regulate pH value, be preferably and transfer to 8-9 with ammonia; Stir, place, obtain deposition; This deposition to white, obtains white crystal with chloroform-methanol (8: 1) washing, is the rutaecarpine of purity about 98%.The ammonia precipitation process after-filtration must be filtrated, and this filtrating is extracted to water inanimate object alkali with chloroform-acetone (6: 1), and organic facies concentrates, dry, and the organic solvent recrystallization obtains the rutaecarpin of purity about 98%.
The present invention proposes a kind of new evaluation and treat the method (the body network is to the recurrence of healthy track) of the action effect of bowelcancer medicine.And develop treatment and intervention that a kind of pharmaceutical composition of being made up of berberine, rutaecarpin and rutaecarpine is used for the colon cancer rat that Dimethylhydrazine brings out.
In order to verify reasonability of the present invention, the applicant has carried out the single factor experiment screening to the extraction and the purifying process of berberine, rutaecarpin, rutaecarpine respectively, and is specific as follows:
The research of one, berberine extraction, purifying process
The method for preparing of berberine generally adopt water or Different concentrations of alcohol extraction, concentrate, concentrated solution is adjusted to acidity with acid, removes impurity; Filtrate then under alkali condition; Separate out berberine,, considered that different organic solvents carries out recrystallization in order to improve the purity of berberine.So the single factor of the present invention is investigated concentration of alcohol, quantity of solvent, extraction time, extraction time, pH value, recrystallization solvent respectively, the purity with extractum rate and berberine is performance assessment criteria respectively.
1, the influence of concentration of alcohol
It is an amount of to get the Rhizoma Coptidis coarse powder, with 5 times of amount ethanol extractions of 30%, 60% and 75% 3 times, extracts 2 hours respectively at every turn; Content with paste-forming rate and berberine is performance assessment criteria; The result finds with 5 times of amount 60% ethanol paste-forming rates the highest, and the content of berberine is the highest, reaches more than 60%.
2, the influence of quantity of solvent
It is an amount of to get the Rhizoma Coptidis coarse powder, with 5,10 and 15 times of amount ethanol extractions of 60% 3 times, extracted 2 hours respectively at every turn, and be performance assessment criteria with the content of paste-forming rate and berberine, the result finds that three various amounts of solvent amounts do not have obvious influence to the result.Confirm as 5 times of volumes so extract quantity of solvent.
3, the influence of extraction time
It is an amount of to get the Rhizoma Coptidis coarse powder, with 5 times of amount ethanol extractions of 60% 3 times, extraction time is respectively 1,2,4, hour; Content with paste-forming rate and berberine is performance assessment criteria; It is that 2 hours paste-forming rates are the highest that the result finds to use extraction time, and the content of berberine is the highest, reaches more than 60%.
4, the influence of extraction time
It is an amount of to get the Rhizoma Coptidis coarse powder; Ethanol with 5 times of amounts 60% extracts respectively 2,3 and 4 times; Extraction time was respectively 2 hours, was performance assessment criteria with the content of paste-forming rate and berberine, and the result finds that the content of paste-forming rate and berberine is suitable when extracting 3 times and 4 times; All greater than the result who extracts 2 times, so extraction time is chosen as 3 times.
5, pH value is investigated
Get the extraction concentrated solution, put coldly, add 1% glacial acetic acid, the adjusting pH value is 5-6, separates out a large amount of depositions, and sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and removes deposition.Filtrating is regulated pH value with calcium hydroxide and is respectively 7.5-8.0,8.5-9.0 and 9.5-10, and when the result finds pH value 8.5-9.0, the yellow acicular crystal amount of separating out maximum, and the purity of berberine is the highest, reaches more than 91%.
6, the investigation of recrystallization solvent
Obtain the yellow crystal thing, use methanol, ethanol and acetone recrystallization respectively, the result finds to use ethyl alcohol recrystallization, and obtaining purity is more than 98%, and single related substance is all less than 0.1% berberine monomeric compound.
According to the result that above-mentioned single factor is investigated, confirm the extraction and the purifying process of berberine: get fresh Rhizoma Coptidis and shine dry grinding, obtain the Rhizoma Coptidis coarse powder, coarse powder is measured 60% ethanol extraction 3 times with 5 times; Each 2 hours, filter merging filtrate; Filtrating is concentrated into 40 ℃, and to measure relative densities be 1.45, puts coldly, adds 1% glacial acetic acid; Separate out a large amount of depositions, sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used ethyl alcohol recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine monomeric compound more than 98%.
Two, the extraction and purification process of rutaecarpin and rutaecarpine
Rutaecarpin and rutaecarpine all are from the Chinese medicine Fructus Evodiae, to extract, and in order to improve yield and purity, adopt a flow process to extract purification 2 monomers; The method for extraction and purification of rutaecarpin and rutaecarpine is similar with berberine; The general employing contained certain density ethanol extraction, and impurity is removed in acid, and alkali is regulated pH value; Obtain rutaecarpin and rutaecarpine, obtain using the organic solvent recrystallization behind the bullion.So the single factor of the present invention is investigated and carried out Different concentrations of alcohol, amount of alcohol added, extraction time, extraction time, pH value, recrystallization solvent respectively and investigate, the purity with extractum rate and rutaecarpin and rutaecarpine is performance assessment criteria respectively.
1, the influence of concentration of alcohol
It is an amount of to get the Fructus Evodiae coarse powder; Respectively with 6 times of amount ethanol extractions of 50%, 60% and 75% 3 times; The each extraction 3 hours is performance assessment criteria with the content of paste-forming rate and rutaecarpin and rutaecarpine, and it is the highest that the result finds to measure 75% ethanol paste-forming rates with 6 times; The content of rutaecarpin is the highest, reaches more than 98%.
3, the influence of quantity of solvent
It is an amount of to get the Fructus Evodiae coarse powder, with 4,6 and 8 times of amount ethanol extractions of 75% 3 times, extracts 3 hours respectively at every turn; Content with paste-forming rate and rutaecarpin and rutaecarpine is performance assessment criteria; The result finds that 6 times of amount 75% ethanol paste-forming rates are the highest, and the content of rutaecarpin is the highest, reaches more than 98%.
3, the influence of extraction time
It is an amount of to get the Fructus Evodiae coarse powder; With 6 times of amount ethanol extractions of 75% 3 times; Extraction time is respectively 2,3,4, hour, be performance assessment criteria with the content of paste-forming rate and rutaecarpin and rutaecarpine, the result finds to use extraction time when being 3 hours and 4 hours; The content of paste-forming rate and rutaecarpin does not have significant change, and the content of rutaecarpin reaches more than 98%.
4, the influence of extraction time
It is an amount of to get the Fructus Evodiae coarse powder; Ethanol with 6 times of amounts 75% extracts respectively 2,3 and 4 times; Extraction time was respectively 3 hours, was performance assessment criteria with the content of paste-forming rate and rutaecarpin, and the result finds that the content of paste-forming rate and rutaecarpin is suitable when extracting 3 times and 4 times; All greater than the result who extracts 2 times, so extraction time is chosen as 3 times.
5, pH value is investigated
Get the extraction concentrated solution, put coldly, add 5% sulphuric acid, the adjusting pH value is 2-2.5, places 6 hours, separates out a large amount of brown depositions, and sedimentation and filtration is removed.Filtrating is regulated pH value with ammonia and is respectively 7-8, and 8-9 and 9-10 stirred 15 minutes, places 4 hours, and when the result found pH value 8-9, precipitation capacity was maximum, and the purity of rutaecarpine is the highest, reaches more than 98%.
6, the investigation of recrystallization solvent
Obtain the bullion of rutaecarpine, use chloroform, methanol, ethanol and acetone recrystallization respectively, the result finds that with chloroform-methanol (8: 1) recrystallization obtaining purity is the rutaecarpine monomeric compound more than 98%.Subsequently, the ammonia precipitation process after-filtration must be filtrated, and to water inanimate object alkali, organic facies is concentrated, dry with chloroform-acetone (6: 1) extraction 3 times for this filtrating, and ethyl alcohol recrystallization obtains the rutaecarpin of purity about 98%.
According to the result that above-mentioned single factor is investigated, confirm the extraction and the purifying process of rutaecarpin and rutaecarpine:, with measuring 75% ethanol extractions 3 times with 6 times after the Fructus Evodiae pulverizing medicinal materials, extracted 3 hours at every turn; 60 ℃ of rotary evaporations of extracting solution are separated out to a small amount of deposition, and concentrate uses 5% sulphuric acid adjusting pH value to be 2-2.5, places 6 hours, obtains the brown insoluble matter; Filter, filtrating is the dilute acidic alcoholic solution, uses ammonia to regulate pH value and is 8-9; Stirred 15 minutes, placed 4 hours, obtain deposition; This deposition to white, obtains white crystal with chloroform-methanol (8: 1) washing, is the rutaecarpine of purity about 98%.The ammonia precipitation process after-filtration must be filtrated, and to water inanimate object alkali, organic facies is concentrated, dry with chloroform-acetone (6: 1) extraction 3 times for this filtrating, and ethyl alcohol recrystallization obtains the rutaecarpin of purity about 98%.
Three, combination of berberine, rutaecarpin and rutaecarpine monomer and different proportion thereof and Zuojin Wan are thought to the influence of colon cancer effect at present; Colorectal cancer be a polygenes, rapid, the multipath process of multistep, mainly can be divided into two kinds of approach: approach is repaired in chromosome instability and mispairing.Wherein relate to methylating of multiple oncogene, antioncogene and mismatch repair gene and gene promoter etc.The chromosome instability approach; Its ultimate principle can be described as: chromosomal unstability causes the inactivation (loss of heterozygosity) and the oncogene active of antioncogene; Thereby cause the generation of colorectal cancer, and follow the developmental sequence of " normal mucosa-adenoma-adenocarcinoma ".
In order to set forth combination of berberine, rutaecarpin and rutaecarpine monomer and different proportion thereof and Zuojin Wan itself action effect to colon cancer; The present invention utilizes Dimethylhydrazine to induce model of colon cancer; To berberine: rutaecarpin: rutaecarpine different proportion compositions (being respectively 1: 8: 1,1: 1: 8,6: 3: 1,7: 2: 1,8: 1: 1); Folk prescription berberine, folk prescription rutaecarpin, folk prescription rutaecarpine and Zuojin Wan carry out comparative study, and to confirm berberine: rutaecarpin: the pharmaceutical composition of rutaecarpine has better effect than said monomer and Zuojin Wan to colon cancer
Adopt the urine of the rat of the intestinal cancer model different time points that gas chromatogram and mass spectrometric hyphenated technique bring out Dimethylhydrazine; Carry out ECF and the analysis of TMS derivatization; Carry out GC-MS and analyze, after the acquisition collection of illustrative plates carried out pretreatment, data utilized Turbomass, Matlab and Simca-P 11.0 to analyze respectively; Obtain the not variation of rat metabolism track on the same group of rat; The result shows: with respect to monomer berberine, rutaecarpin, rutaecarpine and Zuojin Wan, inductive precancerous lesion group of Dimethylhydrazine and cancer group rat give the berberine of proper proportion: rutaecarpin: during the rutaecarpine pharmaceutical composition, the metabolism track of rat obviously returns to healthy rat; Prompting is by berberine: rutaecarpin: the compositions that rutaecarpine is formed can play a role to prophylaxis of colon cancer and treatment through the regulation and control of body network.
Description of drawings
The general flow of Fig. 1 metabolism group research
The GC-MS collection of illustrative plates of Fig. 2 ECF derivatization urine
The GC-MS collection of illustrative plates of Fig. 3 TMS derivatization urine
The PCA figure of Fig. 4, healthy group rat and precancerous lesion group and cancer group rat
PLS-DA figure (the R of Fig. 5, healthy group rat and precancerous lesion group and cancer group rat
2X=0.698; Q
2X (cum)=0.613)
The PCA figure of Fig. 6, administration 10 all berberine, rutaecarpin, rutaecarpine and Zuojin Wan group
The PCA figure of Fig. 7, healthy group rat, precancerous lesion group and administration group rat (administration group ratio is a berberine: rutaecarpin: the ratio of rutaecarpine is 6: 3: 1)
The PCA figure of Fig. 8, healthy group rat, cancer group and administration group (administration group ratio is a berberine: rutaecarpin: the ratio of rutaecarpine is 6: 3: 1) rat
The practical implementation instance
Embodiment 1: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 5 times of amount 30% alcohol reflux 3 times, extracts 2 hours at every turn, filter, and merging filtrate, filtrating is concentrated into 40 ℃, and to measure relative densities be 1.45, puts coldly, adds 1% glacial acetic acid, separates out a large amount of depositions, sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 2: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 5 times of amount 60% alcohol reflux 3 times, extracts 2 hours at every turn, filter, and merging filtrate, filtrating is concentrated into 40 ℃, and to measure relative densities be 1.45, puts coldly, adds 1% glacial acetic acid, separates out a large amount of depositions, sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 3: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 5 times of amount 75% alcohol reflux 3 times, extracts 2 hours at every turn, filter, and merging filtrate, filtrating is concentrated into 40 ℃, and to measure relative densities be 1.45, puts coldly, adds 1% glacial acetic acid, separates out a large amount of depositions, sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 4: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 10 times of amount 60% alcohol reflux 3 times, extracts 2 hours at every turn, filter, and merging filtrate, filtrating is concentrated into 40 ℃, and to measure relative densities be 1.45, puts coldly, adds 1% glacial acetic acid, separates out a large amount of depositions, sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 5: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 15 times of amount 60% alcohol reflux 3 times, extracts 2 hours at every turn, filter, and merging filtrate, filtrating is concentrated into 40 ℃, and to measure relative densities be 1.45, puts coldly, adds 1% glacial acetic acid, separates out a large amount of depositions, sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 6: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 5 times of amount 60% alcohol reflux 3 times, extracts 1 hour at every turn, filter, and merging filtrate, filtrating is concentrated into 40 ℃, and to measure relative densities be 1.45, puts coldly, adds 1% glacial acetic acid, separates out a large amount of depositions, sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 7: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 5 times of amount 60% alcohol reflux 3 times, extracts 4 hours at every turn, filter, and merging filtrate, filtrating is concentrated into 40 ℃, and to measure relative densities be 1.45, puts coldly, adds 1% glacial acetic acid, separates out a large amount of depositions, sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 8: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 5 times of amount 60% alcohol reflux 2 times, extracts 2 hours at every turn, filter, and merging filtrate, filtrating is concentrated into 40 ℃, and to measure relative densities be 1.45, puts coldly, adds 1% glacial acetic acid, separates out a large amount of depositions, sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 9: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 5 times of amount 60% alcohol reflux 4 times, extracts 2 hours at every turn, filter, and merging filtrate, filtrating is concentrated into 40 ℃, and to measure relative densities be 1.45, puts coldly, adds 1% glacial acetic acid, separates out a large amount of depositions, sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 10: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 5 times of amount 60% alcohol reflux 2 times, extracts 2 hours at every turn, filters; Merging filtrate, filtrating are concentrated into 40 ℃ to measure relative densities are 1.45, put coldly, add 1% glacial acetic acid; The adjusting pH value is 5-6, separates out a large amount of depositions, and sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 7.5-8.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 11: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 5 times of amount 60% alcohol reflux 3 times, extracts 2 hours at every turn, filters; Merging filtrate, filtrating are concentrated into 40 ℃ to measure relative densities are 1.45, put coldly, add 1% glacial acetic acid; The adjusting pH value is 5-6, separates out a large amount of depositions, and sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 8.5-9.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 12: the preparation of berberine
It is an amount of to get the Rhizoma Coptidis coarse powder, adds 5 times of amount 60% alcohol reflux 3 times, extracts 2 hours at every turn, filters; Merging filtrate, filtrating are concentrated into 40 ℃ to measure relative densities are 1.45, put coldly, add 1% glacial acetic acid; The adjusting pH value is 5-6, separates out a large amount of depositions, and sedimentation and filtration is removed.It is 3-4 that solution adds concentrated hydrochloric acid adjusting pH value, and placement is spent the night, and obtains yellow acicular crystal, and yellow acicular crystal is used the organic solvent recrystallization, and it is 9.5-10.0 that calcium hydroxide is regulated pH value, and obtaining purity is the berberine more than 98%.
Embodiment 13: the preparation of rutaecarpin and rutaecarpine
With measuring 50% ethanol extractions 3 times with 6 times after the Fructus Evodiae pulverizing medicinal materials, extracted 3 hours at every turn, 60 ℃ of rotary evaporations of extracting solution are separated out to a small amount of deposition, and concentrate uses 5% sulphuric acid adjusting pH value to be 2-2.5; Placed 6 hours, and obtained the brown insoluble matter, filter, filtrating is the dilute acidic alcoholic solution; Use ammonia to regulate pH value and be 8-9, stirred 15 minutes, placed 4 hours, obtain deposition; This deposition to white, obtains white crystal with chloroform-methanol (8: 1) washing, is purity about 98%.The ammonia precipitation process after-filtration must be filtrated, and to water inanimate object alkali, organic facies is concentrated, dry with chloroform-acetone (6: 1) extraction 3 times for this filtrating, and ethyl alcohol recrystallization obtains purity greater than 98% rutaecarpin.
Embodiment 14: the preparation of rutaecarpin and rutaecarpine
With measuring 60% ethanol extractions 3 times with 6 times after the Fructus Evodiae pulverizing medicinal materials, extracted 3 hours at every turn, 60 ℃ of rotary evaporations of extracting solution are separated out to a small amount of deposition, and concentrate uses 5% sulphuric acid adjusting pH value to be 2-2.5; Placed 6 hours, and obtained the brown insoluble matter, filter, filtrating is the dilute acidic alcoholic solution; Use ammonia to regulate pH value and be 8-9, stirred 15 minutes, placed 4 hours, obtain deposition; This deposition to white, obtains white crystal with chloroform-methanol (8: 1) washing, is purity about 98%.The ammonia precipitation process after-filtration must be filtrated, and to water inanimate object alkali, organic facies is concentrated, dry with chloroform-acetone (6: 1) extraction 3 times for this filtrating, and ethyl alcohol recrystallization obtains the rutaecarpin of purity about 98%.
Embodiment 15: the preparation of rutaecarpin and rutaecarpine
With measuring 75% ethanol extractions 3 times with 6 times after the Fructus Evodiae pulverizing medicinal materials, extracted 3 hours at every turn, 60 ℃ of rotary evaporations of extracting solution are separated out to a small amount of deposition, and concentrate uses 5% sulphuric acid adjusting pH value to be 2-2.5; Placed 6 hours, and obtained the brown insoluble matter, filter, filtrating is the dilute acidic alcoholic solution; Use ammonia to regulate pH value and be 8-9, stirred 15 minutes, placed 4 hours, obtain deposition; This deposition to white, obtains white crystal with chloroform-methanol (8: 1) washing, is about 98% rutaecarpine of purity.The ammonia precipitation process after-filtration must be filtrated, and to water inanimate object alkali, organic facies is concentrated, dry with chloroform-acetone (6: 1) extraction 3 times for this filtrating, and ethyl alcohol recrystallization obtains purity greater than 98% rutaecarpin.
Embodiment 16: the preparation of rutaecarpin and rutaecarpine
With measuring 75% ethanol extractions 3 times with 4 times after the Fructus Evodiae pulverizing medicinal materials, extracted 3 hours at every turn, 60 ℃ of rotary evaporations of extracting solution are separated out to a small amount of deposition, and concentrate uses 5% sulphuric acid adjusting pH value to be 2-2.5; Placed 6 hours, and obtained the brown insoluble matter, filter, filtrating is the dilute acidic alcoholic solution; Use ammonia to regulate pH value and be 8-9, stirred 15 minutes, placed 4 hours, obtain deposition; This deposition to white, obtains white crystal with chloroform-methanol (8: 1) washing, is the rutaecarpine of purity about 98%.The ammonia precipitation process after-filtration must be filtrated, and to water inanimate object alkali, organic facies is concentrated, dry with chloroform-acetone (6: 1) extraction 3 times for this filtrating, and ethyl alcohol recrystallization obtains purity greater than 98% rutaecarpin.
Embodiment 17: the preparation of rutaecarpin and rutaecarpine
With measuring 75% ethanol extractions 3 times with 8 times after the Fructus Evodiae pulverizing medicinal materials, extracted 3 hours at every turn, 60 ℃ of rotary evaporations of extracting solution are separated out to a small amount of deposition, and concentrate uses 5% sulphuric acid adjusting pH value to be 2-2.5; Placed 6 hours, and obtained the brown insoluble matter, filter, filtrating is the dilute acidic alcoholic solution; Use ammonia to regulate pH value and be 8-9, stirred 15 minutes, placed 4 hours, obtain deposition; This deposition to white, obtains white crystal with chloroform-methanol (8: 1) washing, is the rutaecarpine of purity about 98%.The ammonia precipitation process after-filtration must be filtrated, and to water inanimate object alkali, organic facies is concentrated, dry with chloroform-acetone (6: 1) extraction 3 times for this filtrating, and ethyl alcohol recrystallization obtains purity greater than 98% rutaecarpin.
Embodiment 18: the preparation of rutaecarpin and rutaecarpine
With measuring 75% ethanol extractions 3 times with 8 times after the Fructus Evodiae pulverizing medicinal materials, extracted 2 hours at every turn, 60 ℃ of rotary evaporations of extracting solution are separated out to a small amount of deposition, and concentrate uses 5% sulphuric acid adjusting pH value to be 2-2.5; Placed 6 hours, and obtained the brown insoluble matter, filter, filtrating is the dilute acidic alcoholic solution; Use ammonia to regulate pH value and be 8-9, stirred 15 minutes, placed 4 hours, obtain deposition; This deposition to white, obtains white crystal with chloroform-methanol (8: 1) washing, is the rutaecarpine of purity about 98%.The ammonia precipitation process after-filtration must be filtrated, and to water inanimate object alkali, organic facies is concentrated, dry with chloroform-acetone (6: 1) extraction 3 times for this filtrating, and ethyl alcohol recrystallization obtains purity greater than 98% rutaecarpin.
Embodiment 19: the preparation of rutaecarpin and rutaecarpine
With measuring 75% ethanol extractions 3 times with 8 times after the Fructus Evodiae pulverizing medicinal materials, extracted 4 hours at every turn, 60 ℃ of rotary evaporations of extracting solution are separated out to a small amount of deposition, and concentrate uses 5% sulphuric acid adjusting pH value to be 2-2.5; Placed 6 hours, and obtained the brown insoluble matter, filter, filtrating is the dilute acidic alcoholic solution; Use ammonia to regulate pH value and be 8-9, stirred 15 minutes, placed 4 hours, obtain deposition; This deposition to white, obtains white crystal with chloroform-methanol (8: 1) washing, is the rutaecarpine of purity about 98%.The ammonia precipitation process after-filtration must be filtrated, and to water inanimate object alkali, organic facies is concentrated, dry with chloroform-acetone (6: 1) extraction 3 times for this filtrating, and ethyl alcohol recrystallization obtains purity greater than 98% rutaecarpin.
Embodiment 20: the preparation of rutaecarpin and rutaecarpine
With measuring 75% ethanol extractions 3 times with 8 times after the Fructus Evodiae pulverizing medicinal materials, extracted 4 hours at every turn, 60 ℃ of rotary evaporations of extracting solution are separated out to a small amount of deposition, and concentrate uses 5% sulphuric acid adjusting pH value to be 2-2.5; Placed 6 hours, and obtained the brown insoluble matter, filter, filtrating is the dilute acidic alcoholic solution; Use ammonia to regulate pH value and be 7-8, stirred 15 minutes, placed 4 hours, obtain deposition; This deposition to white, obtains white crystal with chloroform-methanol (8: 1) washing, is the rutaecarpine of purity about 98%.The ammonia precipitation process after-filtration must be filtrated, and to water inanimate object alkali, organic facies is concentrated, dry with chloroform-acetone (6: 1) extraction 3 times for this filtrating, and ethyl alcohol recrystallization obtains purity greater than 98%, and single impurity is less than 0.1% rutaecarpin.
Embodiment 21: the preparation of rutaecarpin and rutaecarpine
With measuring 75% ethanol extractions 3 times with 8 times after the Fructus Evodiae pulverizing medicinal materials, extracted 4 hours at every turn, 60 ℃ of rotary evaporations of extracting solution are separated out to a small amount of deposition, and concentrate uses 5% sulphuric acid adjusting pH value to be 2-2.5; Placed 6 hours, and obtained the brown insoluble matter, filter; Filtrating is the dilute acidic alcoholic solution, uses ammonia to regulate pH value and is 9-10, stirs 15 minutes; Placed 4 hours, and obtained deposition, this deposition is extremely white with chloroform-methanol (8: 1) washing; Obtain white crystal, about 98% for purity, single impurity is less than 0.1% rutaecarpine.The ammonia precipitation process after-filtration must be filtrated, and to water inanimate object alkali, organic facies is concentrated, dry with chloroform-acetone (6: 1) extraction 3 times for this filtrating, and ethyl alcohol recrystallization obtains purity greater than 98% rutaecarpin.
Embodiment 22 berberine, rutaecarpin and rutaecarpine preparation of compositions
The monomeric compound that extracts according to the method described above was according to 6: 3: 1 (berberine: rutaecarpin: rutaecarpine) prepare the employing method of mixing and become the principal agent of solid mixture as preparation.Adopt the equivalent legal system of progressively increasing to be equipped with tablet (method that comprises wet granule compression tablet, dry granulation tabletting and direct powder compression prepares tablet); The filler principal agent of tablet is microcrystalline Cellulose, lactose, starch; Disintegrating agent is the low substituted hydroxy-propyl methylcellulose; Lubricant is magnesium stearate and Pulvis Talci, and binding agent is 3% polyvidone aqueous solution (adjuvant of tablet comprises above-mentioned part, but is not limited to foregoing); It is encapsulated encapsulated with the system granule that the preparation of capsule adopts certain adjuvant (adjuvant comprises microcrystalline Cellulose, lactose, magnesium stearate, the Pulvis Talci etc.) powder of principal agent adding directly to manage.
Embodiment 23: the metabolism group research of rat urine
23.1 the foundation of colon cancer animal model
It is a kind of sophisticated animal model that Dimethylhydrazine is induced model of colon cancer.230 of Wistar male rats are adopted in this research, are divided into 2 groups at random, and 10 of matched groups are pressed 1.2ml/100g body weight lumbar injection 1mmol/L EDTA, 1 time weekly; DMF lures 220 of cancer groups, presses 25mg/kg body weight lumbar injection DMH, and 1 time weekly, DMH is made into the solution of 5mg/ml with 1mmol/L EDTA facing with preceding, and with 1mol/LNaHCO
3Transfer pH to 6.5-7.0,6 weeks of administration, find lump from pathological section, 11 groups stop the DMF administration.Administration 10 all ill foundations that become amount to 11 groups of judgment models successes.
23.2 animal divides into groups and dosage
(11 groups is the precancerous lesion group to lure the cancer treated animal to be divided into 22 groups; 11 groups is the cancer group); Give respectively and berberine: rutaecarpin: rutaecarpine 1: 8: 1,1: 1: 8,6: 3: 1,7: 2: 1,8: 1: 1; Folk prescription berberine group, folk prescription rutaecarpin group, folk prescription rutaecarpine and Zuojin Wan matched group are according to 5ml/kg body weight lumbar injection mixed solution (mass concentration of mixed solution is 1mg/mL) and positive drug matched group, and 1 time weekly, positive drug is chosen as aspirin; According to 1mg/kg body weight lumbar injection, 1 time weekly; And cox 2 inhibitor (celecoxib) is according to 0.25mg/kg body weight lumbar injection, 1 time weekly; Respectively at giving and pharmaceutical intervention and treatment in 4 weeks after the modeling and 8 weeks.
23.3 the metabolism group research of rat urine
The research flow process of metabolism group in the laboratory animal field generally includes the foundation of the animal model that is fit to early stage, sample collection and preparation, the data analysis in metabolite detection, Analysis and Identification and the later stage in mid-term and modelling and biochemical the explanation.Detailed flow process is seen Fig. 1.
23.3.1 urine collecting
After precancerous lesion group and the modeling of the cancer group rat success, after each administration 8 hours, utilize the rat metabolic cage, one in every cage.Whole urines of collecting 24 hours are used for the research of metabolism group, are stored in-20 ℃ of refrigerators.Carry out Instrumental Analysis until sample.
23.3.2 urine-derived method:
(1) TMS derivatization method: the urine of Dimethylhydrazine rat, with regulating PH with hydrochloric acid and sodium hydroxide behind the acetonitrile precipitation albumen, concentrated, dry, residue is used acetic acid ethyl dissolution, carries out the TMS derivatization behind the ethyl acetate layer nitrogen drying.Concrete operations are following:
A. get the 1ml urine and place teat glass, add the hydrochloric acid of 1ml acetonitrile and 1ml 20% successively, 95 ℃ of water-bath hydrolysis of mixture 1h;
B. the sodium hydroxide 1ml neutralizing hydrolysis thing that in above-mentioned hydrolyzed solution, adds 0.1M, 70 ℃ of water bath methods, residue is used the 2ml acetic acid ethyl dissolution;
C. behind the ethyl acetate solution concussion 10min, the centrifugal 5min of 3000rpm separates upper strata (ethyl acetate layer), and nitrogen dries up;
D. add TMS derivatization reagent 50 μ L, 70 ℃ of derivative reactions 30 minutes.Get 0.5 μ L and inject GC-MS.
(2) ECF derivatization method: urine adds anhydrous ethyl chloroformate, chloroform deposition, and sodium hydroxide is regulated pH value, and the back is ultrasonic, centrifugal, separates chloroform layer, direct injected, and chloroform layer is removed the moisture direct injected.Concrete operations are following:
A. get 300 μ L and place teat glass, add successively urine 300 μ L, interior mark liquid 100 μ L (the L-2-chlorophenylalanine, 0.1mg/mL), dehydrated alcohol 400 μ L, pyridine 100 μ L and ethyl chloroformate 50 μ L, vortex 10s, ultrasonic 1min;
B. add chloroform 300 μ L, vortex 30s, the centrifugal 5min of 3000rpm;
C. add 7MNaOH solution 100 μ L (regulating pH value) and ethyl chloroformate 50 μ L simultaneously, vortex 10s, ultrasonic 1min, vortex 30s, the centrifugal 5min of 3000rpm between the 9-10;
D. the layering solution that obtains is taken out lower floor's chloroform layer, add an amount of anhydrous sodium sulfate Ex-all moisture, therefrom take out and place GC-MS sample introduction bottle about 200 μ L.
23.3.3 GC-MS analyzes
The sample of derivatization under optimized conditions, is carried out GC-MS and analyzes, obtain to analyze collection of illustrative plates (Fig. 2 and Fig. 3) as follows:
23.4 date processing and pattern recognition
Urine is analyzed through derivatization GC-MS; The original collection of illustrative plates that obtains need be through a pre-treatment process; This process mainly comprises the denoising sound, differentiates except that interior mark, peak, and the peak aligns, steps such as normalization; Data through after the pre-treatment can be carried out multidimensional and one-dimensional data analysis, thereby obtain more result and explanation near biological significance.Through Turbomass software declare the peak program with Matlab7.0, processing procedure can be divided into roughly that baseline is corrected, the eliminating at the assorted peak of peak differentiation and coupling, interior mark and some systems and carry out peak normalization with the method for peak area summation.Finally obtain a three-dimensional matrice of forming by the peak intensity after specified peak serial number (corresponding), observation station (sample number) and the normalization with retention time and mass-to-charge ratio; Then; The above-mentioned three-dimensional matrice of handling through Matlab is imported to Simca-P 11.0 software kit (Umetrics;
Sweden) is used for multidimensional statistics and pattern recognition in; In pattern recognition; The metabolism group non-supervision commonly used and the multidimensional statistics method of supervision have been adopted in this research, earlier with the mode identification method-principal component analysis PCA of non-supervision, come assembling naturally, dispersing and outlier of observation sample.And then confirm the network change track situation of its different biological specimens at the different quadrants of pattern recognition.Obtain following result respectively:
We can see from the PCA shot chart, and healthy group rat separates on the PC one dimension obviously with other two groups, and the precancerous lesion group produces on the PC two dimension with the cancer group and separates, can infer tentatively that health organizes metabolism and compose and produced certain variation with respect to model group.
For further proof; Rat precancerous lesion group is after giving certain pharmaceutical intervention; The track of organism metabolism network change, we have carried out the pattern recognition of the administration of rat precancerous lesion group normal healthy controls group, 9 groups of precancerous lesion administration groups and 2 groups of positive drug groups during 6 weeks respectively.When discovery gave berberine, rutaecarpin and the rutaecarpine pharmaceutical composition of rat different proportion, the metabolic profile of rat had regressive trend to the health group.But when giving monomer berberine, rutaecarpin, rutaecarpine and Zuojin Wan group separately; The positive controls of the metabolic profile of rat and 2 chemicalses does not have the significance difference, to the regressive trend of health not obvious (Fig. 6 is seen in the PCA figure variation of berberine, rutaecarpin, rutaecarpine and Zuojin Wan group after 10 weeks of administration).Yet; When giving berberine: rutaecarpin: rutaecarpine is in the time of 6: 3: 1; Certain variation (PCA figure see Fig. 7) has taken place in the metabolic patterns of administration different time points rat, the regressive trend of the 2 oriented health of all metabolism patterns after the rat administration, 4 weeks and 5 weeks certain intersection was arranged after the administration; It is not fairly obvious that this 2 component leaves, but the metabolic profile of 6 all rats returns fairly obvious to the health group after the administration.Explain that rat gives berberine: rutaecarpin: rutaecarpine 6: 3: 1 o'clock, rat have reached one and on whole metabolism network, have more leveled off to healthy balance.Berberine is described: rutaecarpin: the precancerous lesion of 6: 3: 1 pairs of colon cancer of rutaecarpine has tangible intervention effect.
Simultaneously for further proof; The cancer group is after giving certain pharmaceutical intervention; The track of organism metabolism network change, we have carried out the pattern recognition of the administration of rat cancer group normal healthy controls group, 9 groups of cancer group administration groups (comprising monomer berberine, rutaecarpin, rutaecarpine and Zuojin Wan group) and positive drug group during 10 weeks respectively.Discovery gives berberine, rutaecarpin and the rutaecarpine of rat different proportion, and the metabolic profile of rat has regressive trend to the health group.And when giving berberine: rutaecarpin: rutaecarpine is (PCA figure sees Fig. 8) in the time of 6: 3: 1, and the regressive track of rat is similar basically with the regressive track of precancerous lesion group; Certain recurrence trend was arranged after the administration in 2 weeks, and 4 weeks and 6 all metabolism patterns are basic identical after the administration, and 2 groups have no differentiation basically; Rat is described under morbid state, possible body has reached a stable state, yet; In 10 weeks after the administration, rat returns fairly obvious to the health group.Explain that rat gives berberine: rutaecarpin: rutaecarpine 6: 3: 1 o'clock, rat have reached one and on whole metabolism network, have more leveled off to healthy balance.Berberine is described: rutaecarpin: the cancer card of 6: 3: 1 pairs of colon cancer of rutaecarpine has the obvious treatment effect.
Conclusion: show through above experiment; Berberine: rutaecarpin: rutaecarpine 6: 3: 1 o'clock has on metabolic patterns obviously to the regressive trend of health the inductive colon cancer precancerous lesion of Dimethylhydrazine and colon cancer, can explain tentatively that said composition possibly produce through the change to collective's network change to intervene and the effect of treatment colon cancer.
Claims (3)
1. pharmaceutical composition that is used to prevent and/or treat colon cancer; It is characterized in that said active ingredient in pharmaceutical is made up of wherein said berberine: rutaecarpin berberine, rutaecarpin, rutaecarpine: the ratio of rutaecarpine is 6: 3: 1.
2. according to the pharmaceutical composition of claim 1, said compositions exists with the dosage form of tablet, capsule, granule or injection.
3. claim 1 or 2 pharmaceutical composition prevent and/or treat the application in the medicine of colon cancer in preparation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102868557A CN101978962B (en) | 2010-09-20 | 2010-09-20 | Medicinal composition for colon cancer and method for preparing effective monomers thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010102868557A CN101978962B (en) | 2010-09-20 | 2010-09-20 | Medicinal composition for colon cancer and method for preparing effective monomers thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101978962A CN101978962A (en) | 2011-02-23 |
| CN101978962B true CN101978962B (en) | 2012-04-18 |
Family
ID=43599518
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2010102868557A Active CN101978962B (en) | 2010-09-20 | 2010-09-20 | Medicinal composition for colon cancer and method for preparing effective monomers thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101978962B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102608241A (en) * | 2011-11-09 | 2012-07-25 | 涂瑶生 | Traditional Chinese medicine (TCM) active ingredient fingerprint quality control method established on basis of Caco-2 cell model |
| US20150231163A1 (en) * | 2012-08-17 | 2015-08-20 | University-Industry Cooperation Group Of Kyung Hee University | Composition for preventing or treating colitis |
| CN103301134B (en) * | 2013-06-29 | 2014-09-03 | 成都中医药大学 | Pharmaceutical composition for treating hyperlipidemia as well as preparation method and applications |
| CN106892918B (en) * | 2017-01-12 | 2019-02-01 | 南昌大学 | A kind of new method preparing Rutaecarpine from evodia rutaecarpa |
| CN110384680B (en) * | 2018-04-17 | 2021-07-30 | 成都医学院 | temperature/pH responsive double-drug-loading composite nanoparticle and preparation method and application thereof |
| CN109369650A (en) * | 2018-12-17 | 2019-02-22 | 苏州汇通色谱分离纯化有限公司 | The middle method for suppressing standby chromatography and purifying high-purity rutaecarpin and Rutaecarpine |
| CN109942573A (en) * | 2019-04-03 | 2019-06-28 | 海南制药厂有限公司 | A method of separation prepares jamaicin from barberry |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1733097A (en) * | 2005-08-26 | 2006-02-15 | 上海中医药大学 | Application of match drug Chinese goldthread and evodia fruit extracts in preparation of medicine for preventing and treating cancer |
-
2010
- 2010-09-20 CN CN2010102868557A patent/CN101978962B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1733097A (en) * | 2005-08-26 | 2006-02-15 | 上海中医药大学 | Application of match drug Chinese goldthread and evodia fruit extracts in preparation of medicine for preventing and treating cancer |
Non-Patent Citations (3)
| Title |
|---|
| Ming-Lu Xu et al.Cytotoxicity and DNA Topoisomerase Inhibitory Activity of Constituents Isolated from the Fruits of Evodia officinalis.《Archives of Pharmacal Research》.2006,第29卷(第7期),第541-547页. * |
| 常金荣等.吴茱萸碱和小檗碱对HT29细胞凋亡及端粒酶催化亚基( hTERT)表达的影响.《中药药理与临床》.2008,第24卷(第2期),第6-8页. * |
| 董立等.黄连和吴茱萸水提物对大鼠结肠癌癌前病变及结肠上皮增殖和凋亡的影响.《上海中医药杂志》.2010,第44卷(第1期),第66-68页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101978962A (en) | 2011-02-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101978962B (en) | Medicinal composition for colon cancer and method for preparing effective monomers thereof | |
| CN100548323C (en) | A kind of pharmaceutical composition of making by Herb Gynostemmae Pentaphylli, Radix Panacis Quinquefolii and the Radix Astragali | |
| CN102319275B (en) | Sunset abelmoschus flower extract, preparation and preparation method thereof | |
| CN102631390B (en) | Preparation method of periploca forrestii schltr extract as well as product and application of periploca forrestii schltr extract | |
| US10624938B2 (en) | Total flavone extract of flower of abelmoschus manihot L. medic and preparation method thereof | |
| CN101744978B (en) | Drug composition for preventing and curing diabetes mellitus | |
| CN102145062A (en) | Active extracts of rosa roxburghii tratt fruit, and preparation method, detection method and application thereof | |
| CN1965929A (en) | Preparation method of medicament for treating diabetes | |
| CN100574768C (en) | A kind of anticancer pharmaceutical composition and its production and use | |
| CN102327309A (en) | Barberry bark extractive and use of barberry bark extractive and barberry bark | |
| CN101884749B (en) | Particle for eliminating turbidity and treating arthralgia | |
| CN100534493C (en) | Novel antineoplastic compound medicine | |
| CN1961898B (en) | An antitumor compound pharmaceutical composition with barbed stullcap and preparation process thereof | |
| CN101229265B (en) | Antidiabetics and antiatheroscloresis medicine compounds and preparing method thereof | |
| CN103301134B (en) | Pharmaceutical composition for treating hyperlipidemia as well as preparation method and applications | |
| US6726938B2 (en) | Fagopyrum cymosum (Trev.) Meisn composition, method to prepare and analyze the same and uses thereof | |
| CN1954838B (en) | Medical composite of antineoplastic | |
| CN101108214B (en) | Method of separating and extracting natural base from coptis chinensis with latex membrane | |
| CN100482266C (en) | Medical composite prepared by sarcandra and oldenlandia | |
| CN1977886B (en) | Medicinal composition of oxymatrine, ganoderma lucidum and astragalus | |
| CN101049355B (en) | Composition of medication prepared from safflower and leaves of hawthorn | |
| CN103191238A (en) | Fructus aurantii immaturus or fructus aurantii total-flavonoid extract obtained through ethanol reflux extraction and application thereof | |
| CN100493522C (en) | Medicinal composition of oxymatrine and polysaccharide | |
| CN101537086B (en) | Pharmaceutical composition for resisting hepatitis B virus, lowering lipid and alleviating fatty live lesions and preparation method thereof | |
| CN101181403A (en) | Pharmaceutical combination for curing hyperlipemia symptoms and preparation method 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 | ||
| ASS | Succession or assignment of patent right |
Free format text: FORMER OWNER: CHEN PENG CHEN RONGNA Effective date: 20131016 Owner name: BEIJING NUOKANGDA MEDICINE TECHNOLOGY CO., LTD. Free format text: FORMER OWNER: TAO XIUMEI Effective date: 20131016 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 100176 DAXING, BEIJING TO: 100085 HAIDIAN, BEIJING |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20131016 Address after: 100085 B316, B318, 5 Pioneer Road, Beijing, Haidian District Patentee after: BEIJING NUOKANGDA PHARMACEUTICAL CO., LTD. Address before: 100176 No. 6 Garden Street, Beijing economic and Technological Development Zone Patentee before: Tao Xiumei Patentee before: Chen Peng Patentee before: Chen Rongna |
|
| DD01 | Delivery of document by public notice |
Addressee: BEIJING NUOKANGDA PHARMACEUTICAL CO., LTD. Document name: Notification to Pay the Fees |
|
| ASS | Succession or assignment of patent right |
Owner name: INCREASE PHARMACEUTICAL (YINGKOU) CO., LTD. Free format text: FORMER OWNER: BEIJING NUOKANGDA MEDICINE TECHNOLOGY CO., LTD. Effective date: 20150508 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: ADDRESS; FROM: 100085 HAIDIAN, BEIJING TO: 115004 YINGKOU, LIAONING PROVINCE |
|
| TR01 | Transfer of patent right |
Effective date of registration: 20150508 Address after: 115004, No. 97, Donghai Avenue, downtown West, Yingkou, Liaoning Patentee after: Yinkeruisi Pharmaceutical (Yingkou) Co., Ltd. Address before: 100085 B316, B318, Haidian District, Beijing, 5 Patentee before: BEIJING NUOKANGDA PHARMACEUTICAL CO., LTD. |
|
| DD01 | Delivery of document by public notice |
Addressee: Mou Die Document name: Notification of Passing Examination on Formalities |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20160301 Address after: 100085 B316, B318, 5 Pioneer Road, Beijing, Haidian District Patentee after: BEIJING NUOKANGDA PHARMACEUTICAL CO., LTD. Address before: 115004, No. 97, Donghai Avenue, downtown West, Yingkou, Liaoning Patentee before: Yinkeruisi Pharmaceutical (Yingkou) Co., Ltd. |
|
| DD01 | Delivery of document by public notice |
Addressee: BEIJING NUOKANGDA PHARMACEUTICAL CO., LTD. Document name: Notification of Passing Examination on Formalities |
|
| DD01 | Delivery of document by public notice | ||
| DD01 | Delivery of document by public notice |
Addressee: BEIJING NUOKANGDA PHARMACEUTICAL CO., LTD. Document name: Notification to Pay the Fees |
|
| CP03 | Change of name, title or address | ||
| CP03 | Change of name, title or address |
Address after: 100176 Beijing Beijing economic and Technological Development Zone, Chuang Chuang thirteen Street 31 hospital two District 7 Building 101 room. Patentee after: Beijing nukangda medicine Polytron Technologies Inc Address before: 100085 B316, B318, 5 development road, Haidian District, Beijing. Patentee before: BEIJING NUOKANGDA PHARMACEUTICAL CO., LTD. |
|
| DD01 | Delivery of document by public notice | ||
| DD01 | Delivery of document by public notice |
Addressee: Beijing nukangda medicine Polytron Technologies Inc Document name: Notification of Passing Examination on Formalities |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190220 Address after: Room 101-2, 1st floor, 7th floor, 2nd District, 31 Kechuang Thirteenth Street, Beijing Economic and Technological Development Zone, 100176 Patentee after: Beijing Collinmede Pharmaceutical Technology Co., Ltd. Address before: 100176 Beijing Beijing economic and Technological Development Zone, Chuang Chuang thirteen Street 31 hospital two District 7 Building 101 room. Patentee before: Beijing nukangda medicine Polytron Technologies Inc |
|
| DD01 | Delivery of document by public notice | ||
| DD01 | Delivery of document by public notice |
Addressee: Beijing nukangda medicine Polytron Technologies Inc Document name: Notification of Passing Examination on Formalities |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20200723 Address after: Plot 0501-57-4, Daxing biomedical industry base, Zhongguancun Science and Technology Park, Daxing District, Beijing 100176 Patentee after: Beijing Connaught Pharmaceutical Co., Ltd. Address before: Room 101-2, 1st floor, 7th floor, 2nd District, 31 Kechuang Thirteenth Street, Beijing Economic and Technological Development Zone, 100176 Patentee before: Beijing Collinmede Pharmaceutical Technology Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20201217 Address after: Room 1026, No.1, building 14, Daxing Economic Development Zone, Beijing Patentee after: Beijing Renzhong Pharmaceutical Co.,Ltd. Address before: Plot 0501-57-4, Daxing biomedical industry base, Zhongguancun Science and Technology Park, Daxing District, Beijing 100176 Patentee before: Beijing Connaught Pharmaceutical Co., Ltd. |