CN106361731A - Bibenzyl compound, preparation method thereof, and application thereof in preparation of antitumor medicines - Google Patents
Bibenzyl compound, preparation method thereof, and application thereof in preparation of antitumor medicines Download PDFInfo
- Publication number
- CN106361731A CN106361731A CN201610719859.7A CN201610719859A CN106361731A CN 106361731 A CN106361731 A CN 106361731A CN 201610719859 A CN201610719859 A CN 201610719859A CN 106361731 A CN106361731 A CN 106361731A
- Authority
- CN
- China
- Prior art keywords
- eluent
- ethyl acetate
- water
- ethanol
- methanol
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 239000003814 drug Substances 0.000 title claims abstract description 12
- -1 Bibenzyl compound Chemical class 0.000 title description 13
- 229940079593 drug Drugs 0.000 title description 4
- 230000000259 anti-tumor effect Effects 0.000 title 1
- 150000001875 compounds Chemical class 0.000 claims abstract description 77
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 15
- 150000003839 salts Chemical group 0.000 claims abstract description 11
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims abstract description 6
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims abstract description 4
- 239000013078 crystal Chemical group 0.000 claims abstract description 3
- 239000012453 solvate Substances 0.000 claims abstract description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 96
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 78
- 239000003480 eluent Substances 0.000 claims description 71
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 58
- 238000010828 elution Methods 0.000 claims description 55
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 50
- 239000002024 ethyl acetate extract Substances 0.000 claims description 34
- 239000002904 solvent Substances 0.000 claims description 34
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 33
- WGLUMOCWFMKWIL-UHFFFAOYSA-N dichloromethane;methanol Chemical compound OC.ClCCl WGLUMOCWFMKWIL-UHFFFAOYSA-N 0.000 claims description 32
- 239000003208 petroleum Substances 0.000 claims description 31
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims description 28
- 239000000284 extract Substances 0.000 claims description 26
- 239000012530 fluid Substances 0.000 claims description 26
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 19
- 238000010898 silica gel chromatography Methods 0.000 claims description 18
- 238000002156 mixing Methods 0.000 claims description 15
- 239000000469 ethanolic extract Substances 0.000 claims description 13
- 238000012746 preparative thin layer chromatography Methods 0.000 claims description 13
- 229920005654 Sephadex Polymers 0.000 claims description 11
- 239000012507 Sephadex™ Substances 0.000 claims description 11
- 239000000499 gel Substances 0.000 claims description 11
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 9
- 238000004519 manufacturing process Methods 0.000 claims description 9
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 8
- 239000003795 chemical substances by application Substances 0.000 claims description 8
- 238000004440 column chromatography Methods 0.000 claims description 7
- 238000004007 reversed phase HPLC Methods 0.000 claims description 6
- FDSGHYHRLSWSLQ-UHFFFAOYSA-N dichloromethane;propan-2-one Chemical compound ClCCl.CC(C)=O FDSGHYHRLSWSLQ-UHFFFAOYSA-N 0.000 claims description 5
- 201000005202 lung cancer Diseases 0.000 claims description 5
- 208000020816 lung neoplasm Diseases 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 239000011347 resin Substances 0.000 claims description 5
- 229920005989 resin Polymers 0.000 claims description 5
- 239000000741 silica gel Substances 0.000 claims description 5
- 229910002027 silica gel Inorganic materials 0.000 claims description 5
- 239000004793 Polystyrene Substances 0.000 claims description 4
- 201000011510 cancer Diseases 0.000 claims description 4
- 229920002223 polystyrene Polymers 0.000 claims description 4
- 229920005990 polystyrene resin Polymers 0.000 claims description 4
- 238000003820 Medium-pressure liquid chromatography Methods 0.000 claims description 3
- FGUUSXIOTUKUDN-IBGZPJMESA-N C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 Chemical compound C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 FGUUSXIOTUKUDN-IBGZPJMESA-N 0.000 claims description 2
- HWJHWSBFPPPIPD-UHFFFAOYSA-N ethoxyethane;propan-2-one Chemical compound CC(C)=O.CCOCC HWJHWSBFPPPIPD-UHFFFAOYSA-N 0.000 claims description 2
- QWUWMCYKGHVNAV-UHFFFAOYSA-N 1,2-dihydrostilbene Chemical group C=1C=CC=CC=1CCC1=CC=CC=C1 QWUWMCYKGHVNAV-UHFFFAOYSA-N 0.000 abstract description 18
- 210000004027 cell Anatomy 0.000 abstract description 12
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 abstract description 7
- 229910052739 hydrogen Inorganic materials 0.000 abstract description 7
- 239000001257 hydrogen Substances 0.000 abstract description 7
- 210000004881 tumor cell Anatomy 0.000 abstract description 5
- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- 201000005296 lung carcinoma Diseases 0.000 abstract 1
- 235000019441 ethanol Nutrition 0.000 description 24
- 230000002829 reductive effect Effects 0.000 description 12
- 239000000463 material Substances 0.000 description 11
- 239000000243 solution Substances 0.000 description 10
- 238000004809 thin layer chromatography Methods 0.000 description 10
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 239000000203 mixture Substances 0.000 description 8
- 239000000843 powder Substances 0.000 description 8
- 238000001228 spectrum Methods 0.000 description 8
- 230000000694 effects Effects 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 6
- 238000000746 purification Methods 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 238000005160 1H NMR spectroscopy Methods 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- 238000001035 drying Methods 0.000 description 5
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 5
- ROAYSRAUMPWBQX-UHFFFAOYSA-N ethanol;sulfuric acid Chemical compound CCO.OS(O)(=O)=O ROAYSRAUMPWBQX-UHFFFAOYSA-N 0.000 description 5
- 238000000605 extraction Methods 0.000 description 5
- 238000010992 reflux Methods 0.000 description 5
- 238000000926 separation method Methods 0.000 description 5
- 230000003595 spectral effect Effects 0.000 description 5
- 239000012085 test solution Substances 0.000 description 5
- 238000011282 treatment Methods 0.000 description 5
- 239000002246 antineoplastic agent Substances 0.000 description 4
- 229940041181 antineoplastic drug Drugs 0.000 description 4
- 229910052799 carbon Inorganic materials 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 239000002552 dosage form Substances 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical group ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 125000000217 alkyl group Chemical group 0.000 description 3
- 239000002775 capsule Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000008194 pharmaceutical composition Substances 0.000 description 3
- 239000006187 pill Substances 0.000 description 3
- 239000007909 solid dosage form Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 239000003826 tablet Substances 0.000 description 3
- PUPZLCDOIYMWBV-UHFFFAOYSA-N (+/-)-1,3-Butanediol Chemical compound CC(O)CCO PUPZLCDOIYMWBV-UHFFFAOYSA-N 0.000 description 2
- VBICKXHEKHSIBG-UHFFFAOYSA-N 1-monostearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(O)CO VBICKXHEKHSIBG-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 241001313857 Bletilla striata Species 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 235000010419 agar Nutrition 0.000 description 2
- 235000010443 alginic acid Nutrition 0.000 description 2
- 229920000615 alginic acid Polymers 0.000 description 2
- 125000004432 carbon atom Chemical group C* 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 239000003995 emulsifying agent Substances 0.000 description 2
- 239000000839 emulsion Substances 0.000 description 2
- 238000005265 energy consumption Methods 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000009776 industrial production Methods 0.000 description 2
- 239000003701 inert diluent Substances 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000008297 liquid dosage form Substances 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 210000000582 semen Anatomy 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000000375 suspending agent Substances 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000011200 topical administration Methods 0.000 description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- 238000001291 vacuum drying Methods 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- JNYAEWCLZODPBN-JGWLITMVSA-N (2r,3r,4s)-2-[(1r)-1,2-dihydroxyethyl]oxolane-3,4-diol Chemical class OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O JNYAEWCLZODPBN-JGWLITMVSA-N 0.000 description 1
- 229940058015 1,3-butylene glycol Drugs 0.000 description 1
- BRSRUYVJULRMRQ-UHFFFAOYSA-N 1-phenylanthracene Chemical class C1=CC=CC=C1C1=CC=CC2=CC3=CC=CC=C3C=C12 BRSRUYVJULRMRQ-UHFFFAOYSA-N 0.000 description 1
- LUXDHEOYGQPBDS-UHFFFAOYSA-N 2,4-bis[(4-hydroxyphenyl)methyl]-5-methoxy-3-[2-(3-methoxyphenyl)ethyl]phenol Chemical group COC1=CC=CC(CCC=2C(=C(OC)C=C(O)C=2CC=2C=CC(O)=CC=2)CC=2C=CC(O)=CC=2)=C1 LUXDHEOYGQPBDS-UHFFFAOYSA-N 0.000 description 1
- AKPZBCLMSVLMLQ-UHFFFAOYSA-N 2-[(4-hydroxyphenyl)methyl]-3-methoxy-5-[2-(3-methoxyphenyl)ethyl]phenol Chemical group COC1=CC=CC(CCC=2C=C(OC)C(CC=3C=CC(O)=CC=3)=C(O)C=2)=C1 AKPZBCLMSVLMLQ-UHFFFAOYSA-N 0.000 description 1
- AEHPFSANMZYPCC-UHFFFAOYSA-N 2-[(4-hydroxyphenyl)methyl]-5-methoxy-3-[2-(3-methoxyphenyl)ethyl]phenol Chemical group COC1=CC=CC(CCC=2C(=C(O)C=C(OC)C=2)CC=2C=CC(O)=CC=2)=C1 AEHPFSANMZYPCC-UHFFFAOYSA-N 0.000 description 1
- MQWPGJIAXPPAMS-UHFFFAOYSA-N 3-[2-[3-hydroxy-4-[(4-hydroxyphenyl)methyl]phenyl]ethyl]-2,4-bis[(4-hydroxyphenyl)methyl]-5-methoxyphenol Chemical group COc1cc(O)c(Cc2ccc(O)cc2)c(CCc2ccc(Cc3ccc(O)cc3)c(O)c2)c1Cc1ccc(O)cc1 MQWPGJIAXPPAMS-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 208000016557 Acute basophilic leukemia 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
- 239000005995 Aluminium silicate Substances 0.000 description 1
- 206010002198 Anaphylactic reaction Diseases 0.000 description 1
- 235000003276 Apios tuberosa Nutrition 0.000 description 1
- 244000105624 Arachis hypogaea Species 0.000 description 1
- 235000010777 Arachis hypogaea Nutrition 0.000 description 1
- 235000010744 Arachis villosulicarpa Nutrition 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- 235000019739 Dicalciumphosphate Nutrition 0.000 description 1
- 239000004606 Fillers/Extenders Substances 0.000 description 1
- KRHYYFGTRYWZRS-UHFFFAOYSA-N Fluorane Chemical compound F KRHYYFGTRYWZRS-UHFFFAOYSA-N 0.000 description 1
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000206672 Gelidium Species 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-N Hydrogen bromide Chemical compound Br CPELXLSAUQHCOX-UHFFFAOYSA-N 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 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 1
- 240000007472 Leucaena leucocephala Species 0.000 description 1
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 1
- 240000003183 Manihot esculenta Species 0.000 description 1
- 235000016735 Manihot esculenta subsp esculenta Nutrition 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 240000007817 Olea europaea Species 0.000 description 1
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 description 1
- 235000004443 Ricinus communis Nutrition 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- SSZBUIDZHHWXNJ-UHFFFAOYSA-N Stearinsaeure-hexadecylester Natural products CCCCCCCCCCCCCCCCCC(=O)OCCCCCCCCCCCCCCCC SSZBUIDZHHWXNJ-UHFFFAOYSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- DTQVDTLACAAQTR-UHFFFAOYSA-M Trifluoroacetate Chemical compound [O-]C(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-M 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 239000003655 absorption accelerator Substances 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 125000005210 alkyl ammonium group Chemical group 0.000 description 1
- 208000030961 allergic reaction Diseases 0.000 description 1
- 235000012211 aluminium silicate Nutrition 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 230000036783 anaphylactic response Effects 0.000 description 1
- 208000003455 anaphylaxis Diseases 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 125000004429 atom Chemical group 0.000 description 1
- 229940077388 benzenesulfonate Drugs 0.000 description 1
- SRSXLGNVWSONIS-UHFFFAOYSA-M benzenesulfonate Chemical compound [O-]S(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-M 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 102000007478 beta-N-Acetylhexosaminidases Human genes 0.000 description 1
- 108010085377 beta-N-Acetylhexosaminidases Proteins 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 235000019437 butane-1,3-diol Nutrition 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 235000010216 calcium carbonate Nutrition 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- CJZGTCYPCWQAJB-UHFFFAOYSA-L calcium stearate Chemical compound [Ca+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O CJZGTCYPCWQAJB-UHFFFAOYSA-L 0.000 description 1
- 235000013539 calcium stearate Nutrition 0.000 description 1
- 239000008116 calcium stearate Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 150000001721 carbon Chemical group 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 229960000541 cetyl alcohol Drugs 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- NEFBYIFKOOEVPA-UHFFFAOYSA-K dicalcium phosphate Chemical compound [Ca+2].[Ca+2].[O-]P([O-])([O-])=O NEFBYIFKOOEVPA-UHFFFAOYSA-K 0.000 description 1
- 229940038472 dicalcium phosphate Drugs 0.000 description 1
- 229910000390 dicalcium phosphate Inorganic materials 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000008298 dragée Substances 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000002702 enteric coating Substances 0.000 description 1
- 238000009505 enteric coating Methods 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000001727 glucose Nutrition 0.000 description 1
- YQEMORVAKMFKLG-UHFFFAOYSA-N glycerine monostearate Natural products CCCCCCCCCCCCCCCCCC(=O)OC(CO)CO YQEMORVAKMFKLG-UHFFFAOYSA-N 0.000 description 1
- SVUQHVRAGMNPLW-UHFFFAOYSA-N glycerol monostearate Natural products CCCCCCCCCCCCCCCCC(=O)OCC(O)CO SVUQHVRAGMNPLW-UHFFFAOYSA-N 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- BXWNKGSJHAJOGX-UHFFFAOYSA-N hexadecan-1-ol Chemical compound CCCCCCCCCCCCCCCCO BXWNKGSJHAJOGX-UHFFFAOYSA-N 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000003906 humectant Substances 0.000 description 1
- 150000002430 hydrocarbons Chemical group 0.000 description 1
- 125000004435 hydrogen atom Chemical group [H]* 0.000 description 1
- 229920003063 hydroxymethyl cellulose Polymers 0.000 description 1
- 229940031574 hydroxymethyl cellulose Drugs 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 229940049920 malate Drugs 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N malic acid Chemical compound OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- XELZGAJCZANUQH-UHFFFAOYSA-N methyl 1-acetylthieno[3,2-c]pyrazole-5-carboxylate Chemical compound CC(=O)N1N=CC2=C1C=C(C(=O)OC)S2 XELZGAJCZANUQH-UHFFFAOYSA-N 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- CQDGTJPVBWZJAZ-UHFFFAOYSA-N monoethyl carbonate Chemical compound CCOC(O)=O CQDGTJPVBWZJAZ-UHFFFAOYSA-N 0.000 description 1
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 1
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 239000004006 olive oil Substances 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920005862 polyol Polymers 0.000 description 1
- 150000003077 polyols Chemical class 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- 238000011321 prophylaxis Methods 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 150000003856 quaternary ammonium compounds Chemical class 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000008159 sesame oil Substances 0.000 description 1
- 235000011803 sesame oil Nutrition 0.000 description 1
- 150000004760 silicates Chemical class 0.000 description 1
- RMAQACBXLXPBSY-UHFFFAOYSA-N silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 229940095064 tartrate Drugs 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 238000001269 time-of-flight mass spectrometry Methods 0.000 description 1
- UAEJRRZPRZCUBE-UHFFFAOYSA-N trimethoxyalumane Chemical compound [Al+3].[O-]C.[O-]C.[O-]C UAEJRRZPRZCUBE-UHFFFAOYSA-N 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/075—Ethers or acetals
- A61K31/085—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/075—Ethers or acetals
- A61K31/085—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon
- A61K31/09—Ethers or acetals having an ether linkage to aromatic ring nuclear carbon having two or more such linkages
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
- C07C41/01—Preparation of ethers
- C07C41/34—Separation; Purification; Stabilisation; Use of additives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
- C07C41/01—Preparation of ethers
- C07C41/34—Separation; Purification; Stabilisation; Use of additives
- C07C41/36—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
- C07C41/01—Preparation of ethers
- C07C41/34—Separation; Purification; Stabilisation; Use of additives
- C07C41/38—Separation; Purification; Stabilisation; Use of additives by liquid-liquid treatment
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/898—Orchidaceae (Orchid family)
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses an application of a compound represented as the formula (IV), or pharmaceutically acceptable salts, crystal forms and solvates thereof, in preparation of a medicine for treating and/or preventing tumors, wherein R1, R4 and R6 are respectively and independently selected from hydrogen or a methyl group; R2, R3, R5 and R7 are respectively and independently selected from hydrogen or a structure represented as follows. The invention provides a novel application of bibenzyl compounds, and in particular, an application in preparation of a medicine for treating and/or preventing tumors. The compound has excellent inhibiting effects on tumor cells, especially, on lung carcinoma cells, wherein IC50 of the compound reaches 2-6 [mu]g/ml. The invention also provides a preparation method of the bibenzyl compounds.
Description
Technical Field
The invention relates to bibenzyl compounds, a preparation method thereof and application thereof in preparing antitumor drugs.
Background
The prior literature reports that bibenzyl compounds have the drug effect and the application of inhibiting passive skin allergic reaction (passive cutaneous anaphylaxis) and beta-hexosaminidase released by RBL-2H3 cells (basophilic leukemia cells) (see: Matsuda H et al. antibacterial phenyl anthracenes and … Planta Med 2004; 70: 847-855):
at present, no research report on the application of the bibenzyl compound in preparing antitumor drugs is found.
Disclosure of Invention
The invention aims to provide a new application of bibenzyl compounds shown as follows, in particular to an application in preparing medicaments for treating and/or preventing tumors.
The invention provides an application of a compound shown in a formula IV or pharmaceutically acceptable salt, crystal form and solvate thereof in preparing a medicament for treating and/or preventing tumors, wherein the compound is as follows:
wherein,
R1、R4、R6each independently selected from H or methyl;
R2、R3、R5、R7each independently selected from H or
The compound shown in the formula IV is:
further, the tumor is cancer; further, the cancer is lung cancer.
The present invention also provides a process for preparing compound F, comprising the steps of:
a. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
b. taking the ethanol extract, and concentrating to obtain a fluid extract;
c. dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, mixing ethyl acetate parts, and recovering solvent to obtain ethyl acetate extract;
d. taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially carrying out gradient elution by using petroleum ether-acetone as an eluent at a ratio of 100:1, at a ratio of 50:1 and at a ratio of 30:1 to obtain petroleum ether: eluent fr.16 when acetone is 30: 1;
e. fr.16 is loaded on a reversed-phase polystyrene resin column, gradient elution is carried out by using ethanol-water of 10:90, 30:70, 50:50, 70:30 and 90:10 as eluent in sequence, and Fr.16-h of the ethanol-water of 90:10 is obtained;
f. fr.16-h is subjected to medium-pressure liquid chromatography, and gradient elution is sequentially carried out by using methanol-water of 20:80, 30:70 and 50:50 as eluent, so as to obtain Fr.16-h-3 when the methanol-water of 50: 50;
g. fr.16-h-3 was chromatographed on silica gel sequentially with dichloromethane-methanol ═ 1: 0. 100, and (2) a step of: 1 as eluent, eluting 4 column volumes respectively, collecting dichloromethane-methanol as 100: fr.16-h-3b at 1;
h. fr.16-h-3 was subjected to gel column chromatography eluting 4 column volumes with methanol-water 70:30 as eluent, followed by preparative thin layer chromatography eluting with dichloromethane-methanol 30:1 as developing agent, RfCompound F was isolated as 0.58.
Further, in the above-mentioned case,
in the step d, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step e, the amount of Fr.16 is 15-20 g, and the gradient elution conditions are as follows:
in the step f, the amount of Fr.16-h is 4-6 g, and the gradient elution conditions are as follows:
the present invention also provides a process for preparing compound G or compound H, comprising the steps of:
i. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
ii. Taking the ethanol extract, and concentrating to obtain a fluid extract;
iii, dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, combining ethyl acetate parts, and recovering the solvent to obtain an ethyl acetate extract;
iv, taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially mixing petroleum ether: gradient elution is carried out by using acetone as an eluent at the ratio of 100:1, 50:1 and 30:1, and petroleum ether is obtained: eluent fr.16 when acetone is 30: 1;
v, fr.16 on a reversed-phase polystyrene type resin column, sequentially mixing ethanol: gradient elution is carried out by using water as eluent, wherein the water is 10:90, 30:70, 50:50, 70:30, 90:10 and 100:0, and ethanol is obtained: fr.16-j when water is 100: 0;
vi, subjecting the Fr.16-j to a sephadex chromatographic column, and sequentially eluting by using dichloromethane-methanol (1: 1) as an eluent, wherein the eluent has the volume of 0.8 time of the column volume and 2.2 times of the column volume to obtain Fr.16-j-1 and Fr.16-j-2 respectively;
vii and fr.16-j-2, silica gel column chromatography, sequentially eluting with dichloromethane-methanol ═ 1: 0. 200: eluting with eluent 1, eluting 3 column volumes respectively, and collecting dichloromethane-methanol (200): fr.16-j-2b at 1;
viii, and Fr.16-j-2b are subjected to sephadex chromatographic column, and petroleum ether-dichloromethane-methanol 2:2:1 is used as eluent, and 1 column volume are sequentially eluted to obtain Fr.16-j-2bA and Fr.16-j-2 bB;
fr.16-j-2bA by preparative thin layer chromatography with dichloromethane-acetone 20:1 as developing solvent, Rf(iv) isolating as 0.76 to give compound G;
fr.16-j-2bB by preparative thin layer chromatography using dichloromethane-acetone 20:1 as developing solvent, Rf0.80, isolated to give compound H.
Further, in the above-mentioned case,
in the step iv, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step v, the amount of Fr.16 is 15-20 g, and gradient elution conditions are as follows:
the present invention also provides a process for preparing compound I, comprising the steps of:
A. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
B. taking the ethanol extract, and concentrating to obtain a fluid extract;
C. dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, mixing ethyl acetate parts, and recovering solvent to obtain ethyl acetate extract;
D. taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially mixing petroleum ether: gradient elution is carried out by using acetone as eluent, wherein the acetone is 100:1, 50:1, 30:1 and 20:1, and petroleum ether is obtained: eluent fr.18 when acetone is 20: 1;
E. fr.18 on a reversed-phase polystyrene type resin column, sequentially eluting with ethanol: gradient elution is carried out by using water as eluent and water as eluent, wherein the water is 40:60, 50:50, 55:45, 60:40, 70:30, 80:20 and 95:5 are carried out to obtain ethanol: fr.18-n at 95: 5;
F. sequentially eluting with dichloromethane-methanol (1: 1) as eluent by Fr.18-n sephadex chromatographic column to obtain Fr.18-n-1, Fr.18-n-2 and Fr.18-n-3 respectively;
fr.18-n-3 was chromatographed on Sephadex column with methanol-water 70:30 as eluent for 3 column volumes, the main compound was chromatographed on preparative thin layer chromatography with dichloromethane-methanol 20:1 as developing solvent, RfThen purified by reverse phase high performance liquid chromatography with methanol-water 65:35 as the mobile phase to give compound I.
Further, in the above-mentioned case,
in the step D, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step E, the amount of Fr.18 is 18-20 g, and the gradient elution conditions are as follows:
the present invention also provides a process for preparing compound J, comprising the steps of:
I. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
II. Taking the ethanol extract, and concentrating to obtain a fluid extract;
III, dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, combining ethyl acetate parts, and recovering the solvent to obtain an ethyl acetate extract;
IV, taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially mixing petroleum ether: gradient elution is carried out by using acetone as eluent, wherein the acetone is 100:1, 50:1, 30:1 and 20:1, and petroleum ether is obtained: eluent fr.18 when acetone is 20: 1;
v, Fr.18 on a reversed-phase polystyrene resin column, sequentially adding ethanol: gradient elution is carried out by using water as eluent and water as eluent, wherein the water is 40:60, 50:50, 55:45, 60:40, 70:30 and 80:20, and ethanol is obtained: fr.18-m at 80: 20;
VI and Fr.18-m are put on a sephadex chromatographic column, and are sequentially eluted by 1 time of column volume and 1 time of column volume by using methanol-water (60: 40) as an eluent to respectively obtain Fr.18-m-1 and Fr.18-m-2;
fr.18-m-2 byEluting with dichloromethane-methanol (1: 1) as eluent for 4 column volumes, performing preparative thin layer chromatography with dichloromethane-methanol (15: 1) as developing agent, and collecting eluate, and separating with chromatographic columnfThen purified by reverse phase high performance liquid chromatography with methanol-water 55:45 as the mobile phase to give compound J.
Further, in the above-mentioned case,
in the step IV, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step V, the amount of Fr.18 is 18-20 g, and the gradient elution conditions are as follows:
the invention provides a new application of bibenzyl compounds, in particular to an application in preparing medicaments for treating and/or preventing tumors, which has good inhibition effect on tumor cells, particularly lung cancer cells and IC thereof502-6 mug/mL; meanwhile, the preparation method of the compound is simple and convenient, mild in reaction conditions, convenient to operate and control, low in energy consumption, high in yield, low in cost and very suitable for industrial production.
The compounds and derivatives provided in the present invention may be named according to the IUPAC (international union of pure and applied chemistry) or CAS (chemical abstracts service, Columbus, OH) naming system.
Definitions of terms used in connection with the present invention: the initial definitions provided herein for a group or term apply to that group or term throughout the specification unless otherwise indicated; for terms not specifically defined herein, the meanings that would be given to them by a person skilled in the art are to be given in light of the disclosure and the context.
"substituted" means that a hydrogen atom in a molecule is replaced by a different atom or molecule.
The minimum and maximum values of the carbon atom content in the hydrocarbon group are indicated by a prefix, e.g. prefix Ca～CbAlkyl means any alkyl group containing "a" to "b" carbon atoms. Thus, for example, C1～C4The alkyl group means an alkyl group containing 1 to 4 carbon atoms, in other words, C1～C4Alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, tert-butyl.
The term "pharmaceutically acceptable" means that the carrier, cargo, diluent, adjuvant, and/or salt formed is generally chemically or physically compatible with the other ingredients comprising a pharmaceutical dosage form and physiologically compatible with the recipient.
The terms "salt" and "pharmaceutically acceptable salt" refer to acid and/or base salts of the above compounds or stereoisomers thereof, with inorganic and/or organic acids and bases, as well as zwitterionic (inner) salts, and also quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly in the final isolation and purification of the compounds. The compound or a stereoisomer thereof may be obtained by appropriately (e.g., equivalently) mixing the above compound or a stereoisomer thereof with a predetermined amount of an acid or a base. These salts may form precipitates in the solution which are collected by filtration, or they may be recovered after evaporation of the solvent, or they may be prepared by reaction in an aqueous medium followed by lyophilization. The salt in the invention can be hydrochloride, sulfate, citrate, benzene sulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, succinate, fumarate, maleate, tartrate or trifluoroacetate of the compound.
The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, parenteral (intravenous, intramuscular, or subcutaneous), and topical administration.
Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.
Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.
Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like.
In addition to these inert diluents, the compositions can also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.
Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.
Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.
Dosage forms for topical administration of the compounds of the present invention include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.
The pharmaceutically acceptable auxiliary material of the invention refers to a substance contained in a dosage form except for an active ingredient.
The pharmaceutically acceptable auxiliary components have certain physiological activity, but the addition of the components does not change the dominant position of the pharmaceutical composition in the disease treatment process, but only plays auxiliary effects, and the auxiliary effects are only the utilization of the known activity of the components and are auxiliary treatment modes which are commonly used in the field of medicine. If the auxiliary components are used together with the pharmaceutical composition of the present invention, the protection scope of the present invention still remains.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Detailed Description
The raw materials and equipment used in the embodiment of the present invention are known products and obtained by purchasing commercially available products.
Firstly, medicinal materials
Bletilla striata medicinal material is purchased from Nenjiang city of Sichuan province at 8 months in 2012, and is identified as a dry tuber of Bletilla striata (Thunb.) Rchb.f. by professor Limin of Chengdu Chinese medicine university.
② reagents and fillers
Column chromatography silica gel of 200-300 mesh (reagent grade) purchased from Qingdao ocean silica gel desiccant factory;
thin layer chromatography silica gel G, GF254 and H (chemically pure), purchased from Qingdao ocean silica gel desiccant factory;
MCI gel CHP 20P, 75-150 μm, is a reversed-phase polystyrene-type resin, and is purchased from Mitsubishi chemical corporation of Japan;
sephadex LH-20 sepharose, available from Amersham, Sweden;
a GF254 silica gel preparation thin layer purchased from Yangtze river friend silica gel development Co., Ltd;
chromatographic methanol, 4L/bottle, available from Fisher corporation, USA;
analytically pure reagents such as petroleum ether, ethyl acetate, n-butanol, acetone, methanol and the like are purchased from Chengdu Kelong chemical reagent factories.
(iii) laboratory instruments
Cometro 6000 high performance liquid chromatography (Cometro, usa);
waters Synapt G2HDMS high resolution time-of-flight mass spectrometry (Waters, usa);
nuclear magnetic resonance apparatus: bruker-600 NMR; measuring temperature: 297K; taking a solvent peak signal as a reference;
vector 22FT-IR infrared spectrometer (Bruker, Switzerland);
shimadzu UV-260 UV-visible spectrophotometer (Shimadzu, japan);
BP211D one tenth ten million electronic balance (Sartorius, switzerland);
r-210 rotary evaporator (BUCHI, Switzerland);
model DZG-6050 vacuum drying oven (shanghai semen).
Example 1 extraction and Structure identification of Compound F
(1) Extracting medicinal materials:
extracting dried rhizoma Bletillae coarse powder (3kg) with 95% ethanol under reflux for 3 times (30L × 3), each time for 2 hr;
(2) separation and purification of components:
firstly, concentrating and drying the ethanol extract (90L) under reduced pressure to obtain 510g of semisolid fluid extract;
dispersing the semi-solid fluid extract (500g) with water (5L), sequentially extracting with ethyl acetate and n-butanol (20L), combining ethyl acetate parts, and recovering the solvent under reduced pressure to obtain 160g of ethyl acetate extract;
③ separating the ethyl acetate extract (160g) by silica gel column chromatography, and purifying by using petroleum ether: gradient elution is carried out on acetone, and the eluent is detected by thin layer chromatography to obtain petroleum ether: eluent fr.16 when acetone is 30: 1;
the conditions for gradient elution were as follows:
processing the Fr.16 component (18g) by MCI, performing gradient elution by using 10-100% v/v ethanol water, and detecting by combining thin-layer chromatography to respectively obtain 10 components Fr.16-a-Fr.16-j;
the conditions for gradient elution were as follows:
fifthly, subjecting the Fr.16-h component (5.0g) to medium pressure liquid chromatography (a B-chi Gradient Former B-687 medium pressure liquid chromatograph) to Gradient elution with 20%, 30%, 50%, 70%, 80%, 95% and 100% methanol water, and eluting 2, 3, 4 and 5 column volumes respectively to obtain seven components Fr.16-h-1-Fr.16-h-7;
sixthly, subjecting the Fr.16-h-3 component (800mg) to silica gel column chromatography, and purifying with a dichloromethane-methanol 1: 0. 100, and (2) a step of: 1. 50: eluent is 1, 4 column volumes are eluted by eluent in each proportion, and 100:1 fraction fr.16-h-3b eluted;
seventhly, carrying out gel column chromatography on the Fr.16-h-3b component (140mg), eluting by 4 column volumes by using 70% methanol water as an eluent, and separating by adopting preparative thin layer chromatography (a developing agent is dichloromethane-methanol (30: 1), and Rf (0.58)) to obtain a compound F;
when the bibenzyl compounds in the rhizoma bletillae are developed by thin-layer chromatography, 10% sulfuric acid ethanol test solution is sprayed, and the rhizoma bletillae is developed for 3min at 105 ℃ to show more yellow, orange to reddish spots, so that the rhizoma bletillae can be used for tracking detection of bibenzyl;
mass spectral data for compound F: ESI-MS M/z 349[ M-H]-；
Hydrogen spectrum data:1H NMR(CD3COCD3,600MHz):7.06(1H,t,J＝7.8Hz,H-5′),6.93(2H,d,J＝8.4Hz,H-2″,6″),6.68(2H,d,J＝8.4Hz,H-3″,5″),6.66(1H,t,J＝1.8Hz,H-2′),6.64(1H,dd,J＝7.8,1.8Hz,H-4′),6.62(1H,dd,J＝7.8,1.8Hz,H-2′),6.40(1H,d,J＝2.4Hz,H-4),6.38(1H,d,J＝2.4Hz,H-6),3.89(2H,s,CH2-γ),3.75(3H,s,OCH3-3),2.59(2H,m,CH2-α),2.74(2H,m,CH2-β)；
carbon spectrum data:13C NMR(CD3COCD3,150MHz):160.2(C-3),158.7(C-3′),158.0(C-5),156.5(C-4″),145.0(C-1′),143.7(C-1),134.0(C-1″),130.6(C-5′),130.3(C-2″,6″),120.8(C-6′),119.8(C-2),116.6(C-4′),116.2(C-3″,5″),114.1(C-2′),109.5(C-6),98.3(C-4),56.3(C-OMe-3),38.6(C-β),36.5(C-α),30.7(C-γ)；
from the above data, compound F was identified as 2-p-hydroxybenzyl-3', 5-dihydroxy-3-methoxybenzyl, and its structure was as follows:
example 2 extraction and Structure identification of Compound G
(1) Extracting medicinal materials:
extracting dried rhizoma Bletillae coarse powder (3kg) with 95% ethanol under reflux for 3 times (30L × 3), each time for 2 hr;
(2) separation and purification of components:
firstly, concentrating and drying the ethanol extract (90L) under reduced pressure to obtain 510g of semisolid fluid extract;
dispersing the semi-solid fluid extract (500g) with water (5L), sequentially extracting with ethyl acetate and n-butanol (20L), combining ethyl acetate parts, and recovering the solvent under reduced pressure to obtain 160g of ethyl acetate extract;
③ separating the ethyl acetate extract (160g) by silica gel column chromatography, and purifying by using petroleum ether: gradient elution is carried out on acetone, and the eluent is detected by thin layer chromatography to obtain petroleum ether: eluent fr.16 when acetone is 30: 1;
the conditions for gradient elution were as follows:
processing the Fr.16 component (18g) by MCI, performing gradient elution by using 10-100% v/v ethanol water, and detecting by combining thin-layer chromatography to respectively obtain 10 components Fr.16-a-Fr.16-j;
the conditions for gradient elution were as follows:
fifthly, enabling the Fr.16-j component (3.0g) to pass through a gel LH-20 chromatographic column, eluting by using dichloromethane-methanol (1: 1) as an eluent, and sequentially collecting the Fr.16-j-1, Fr.16-j-2 and Fr.16-j-3 which are respectively used for eluting 0.8 times of column volume, 2.2 times of column volume and 3 times of column volume, wherein the Fr.16-j-2 component is a concentrated part of the bibenzyl compound;
sixthly, passing the Fr.16-j-2 component (1.8g) through silica gel column chromatography, and respectively carrying out reaction on the components in a weight ratio of dichloromethane-methanol 1: 0. 200: 1. 100, and (2) a step of: 1. 50:1, eluting 3 column volumes by eluent with each proportion, recovering the solvent under reduced pressure, and collecting 200:1 elution parts Fr.16-j-2 b;
⑦ and Fr.16-j-2b fraction (0.8g) passing through gel LH-20 chromatographic column, eluting with petroleum ether-dichloromethane-methanol 2:2:1 as eluent, sequentially eluting for 1 and 1 column volumes to obtain Fr.16-j-2bA and Fr.16-j-2bB fractions, and subjecting Fr.16-j-2bA to thin layer chromatography (developing agent is dichloromethane)Alkyl-propanone ═ 20:1, Rf0.76) to yield compound G;
when the bibenzyl compounds in the rhizoma bletillae are developed by thin-layer chromatography, 10% sulfuric acid ethanol test solution is sprayed, and the rhizoma bletillae is developed for more than yellow, orange to reddish spots at 105 ℃ for 3min, so that the rhizoma bletillae can be used for tracking detection of bibenzyl;
mass spectral data for compound G: ESI-MS M/z 363[ M-H ]]-；
Hydrogen spectrum data:1H NMR(CD3COCD3,600MHz):7.15(1H,t,J＝7.8Hz,H-5′),7.01(2H,d,J＝8.4Hz,H-2″,6″),6.74(1H,brd,J＝7.8Hz,H-4′),6.72(1H,brd,J＝7.8Hz,H-6′)6.71(2H,d,J＝8.4Hz,H-3″,5″),6.70(1H,brs,H-2′),6.41(1H,d,J＝2.4Hz,H-4),6.38(1H,d,J＝2.4Hz,H-6),3.95(2H,s,CH2-γ),3.75(3H,s,OCH3-3′),3.71(3H,s,OCH3-5),2.79(2H,m,CH2-α),2.65(2H,m,CH2-β)；
from the above data, compound G was identified as 2-p-hydroxybenzyl-3-hydroxy-3', 5-dimethoxybibenzyl, having the following structure:
example 3 extraction and Structure identification of Compound H
(1) Extracting medicinal materials:
extracting dried rhizoma Bletillae coarse powder (3kg) with 95% ethanol under reflux for 3 times (30L × 3), each time for 2 hr;
(2) separation and purification of components:
firstly, concentrating and drying the ethanol extract (90L) under reduced pressure to obtain 510g of semisolid fluid extract;
dispersing the semi-solid fluid extract (500g) with water (5L), sequentially extracting with ethyl acetate and n-butanol (20L), combining ethyl acetate parts, and recovering the solvent under reduced pressure to obtain 160g of ethyl acetate extract;
③ separating the ethyl acetate extract (160g) by silica gel column chromatography, and purifying by using petroleum ether: gradient elution is carried out on acetone, and the eluent is detected by thin layer chromatography to obtain petroleum ether: eluent fr.16 when acetone is 30: 1;
the conditions for gradient elution were as follows:
processing the Fr.16 component (18g) by MCI, performing gradient elution by using 10-100% v/v ethanol water, and detecting by combining thin-layer chromatography to respectively obtain 10 components Fr.16-a-Fr.16-j;
the conditions for gradient elution were as follows:
fifthly, enabling the Fr.16-j component (3.0g) to pass through a gel LH-20 chromatographic column, eluting by using dichloromethane-methanol (1: 1) as an eluent, and sequentially collecting the Fr.16-j-1, Fr.16-j-2 and Fr.16-j-3 which are respectively used for eluting 0.8 times of column volume, 2.2 times of column volume and 3 times of column volume, wherein the Fr.16-j-2 component is a concentrated part of the bibenzyl compound;
sixthly, passing the Fr.16-j-2 component (1.8g) through silica gel column chromatography, and respectively carrying out reaction on the components in a weight ratio of dichloromethane-methanol 1: 0. 200: 1. 100, and (2) a step of: 1. 50:1, eluting 3 column volumes by eluent with each proportion, recovering the solvent under reduced pressure, and collecting 200:1 elution parts Fr.16-j-2 b;
⑦ and Fr.16-j-2b fraction (0.8g) are passed through gel LH-20 chromatographic column, eluting with petroleum ether-dichloromethane-methanol 2:2:1 as eluent, sequentially eluting for 1 and 1 column volumes to obtain Fr.16-j-2bA and Fr.16-j-2bB fractions, and Fr.16-j-2bB to prepare thin gelLayer chromatography (developing solvent dichloromethane-acetone ═ 20:1, R)f0.80) to yield compound H;
when the bibenzyl compounds in the rhizoma bletillae are developed by thin-layer chromatography, 10% sulfuric acid ethanol test solution is sprayed, and the rhizoma bletillae is developed for more than yellow, orange to reddish spots at 105 ℃ for 3min, so that the rhizoma bletillae can be used for tracking detection of bibenzyl;
mass spectral data for compound H: ESI-MS M/z 363[ M-H ]]-；
Hydrogen spectrum data:1H NMR(CD3COCD3,600MHz):7.17(1H,t,J＝7.8Hz,H-5′),7.10(2H,d,J＝8.4Hz,H-2″,6″),6.79(1H,d,J＝7.8Hz,H-6′),6.76(1H,brs,H-2′),6.73(1H,dd,J＝7.8,1.8Hz,H-4′),6.66(2H,d,J＝8.4Hz,H-3″,5″),6.41(1H,brs,H-6),6.38(1H,brs,H-2),3.84(2H,s,CH2-γ),3.75(3H,s,OCH3-3′),3.72(3H,s,OCH3-3),2.86(2H,m,CH2-α),2.79(2H,m,CH2-β)；
carbon spectrum data:13C NMR(CD3COCD3,150MHz):160.6(C-3′),159.3(C-3),156.3(C-5),155.9(C-4″),144.4(C-1′),141.8(C-1),133.7(C-1″),130.3(C-2″,6″),130.0(C-5′),121.5(C-6′),115.4(C-3″,5″),114.9(C-2′),114.9(C-4),112.1(C-4′),109.2(C-6),103.6(C-2),55.8(OCH3-3′),55.3(OCH3-3),38.6(CH2-α),38.4(CH2-β),28.3(CH2-γ)；
from the above data, compound H was determined to be 4-p-hydroxybenzyl-5-hydroxy-3, 3' -dimethoxybibenzyl, having the following structure:
example 4 extraction and Structure identification of Compound I
(1) Extracting medicinal materials:
extracting dried rhizoma Bletillae coarse powder (3kg) with 95% ethanol under reflux for 3 times (30L × 3), each time for 2 hr;
(2) separation and purification of components:
firstly, concentrating and drying the ethanol extract (90L) under reduced pressure to obtain 510g of semisolid fluid extract;
dispersing the semi-solid fluid extract (500g) with water (5L), sequentially extracting with ethyl acetate and n-butanol (20L), combining ethyl acetate parts, and recovering the solvent under reduced pressure to obtain 160g of ethyl acetate extract;
③ separating the ethyl acetate extract (160g) by silica gel column chromatography, and purifying by using petroleum ether: gradient eluting with acetone, detecting the eluate by thin layer chromatography, and mixing similar components to obtain petroleum ether: eluent fr.18 when acetone is 20: 1;
the conditions for gradient elution were as follows:
(iv) subjecting the Fr.18 fraction (19g) to MCI treatment, eluting with 40% -100% ethanol water, tracing according to thin layer, and collecting
Recovering solvent from the eluate containing the target compound to obtain 15 fractions Fr.18-a to Fr.18-o;
fifthly, enabling the Fr.18-n component (95% ethanol elution part, 0.7g) to pass through gel LH-20, eluting with dichloromethane-methanol (1: 1) as an eluent, and sequentially collecting the Fr.18-n-1, Fr.18-n-2 and Fr.18-n-3 which are eluted by 1.5 times of column volume, 3 times of column volume and 5 times of column volume respectively;
fr.18-n-3 fraction (300mg) was coagulatedGel LH-20 column chromatography with 70% methanol as eluent, eluting for 3 column volumes, subjecting the main compound to preparative thin layer chromatography (dichloromethane-methanol 20:1 as developing solvent, R)f0.78) and reversed-phase high performance liquid chromatography (65% methanol water is used as a mobile phase) to prepare and purify the compound I;
when the bibenzyl compounds in the rhizoma bletillae are developed by thin-layer chromatography, 10% sulfuric acid ethanol test solution is sprayed, and the rhizoma bletillae is developed for more than yellow, orange to reddish spots at 105 ℃ for 3min, so that the rhizoma bletillae can be used for tracking detection of bibenzyl;
mass spectral data for compound I: ESI-MS M/z 469[ M-H]-；
Hydrogen spectrum data:1H NMR(CD3COCD3,600MHz):7.14(1H,t,J＝7.8Hz,H-5′),7.01(2H,d,J＝8.4Hz,H-H-2″,6″),6.94(2H,d,J＝8.4Hz,H-H-2′,6′),6.67-6.71(6H,m,H-4′,6′,3′,5′,3″,5″),6.63(1H,brs,H-2′),6.61(1H,s,H-4),4.03(2H,s,CH2-γ1),3.98(2H,s,CH2-γ2),3.76(3H,s,OCH3-3′),3.75(3H,s,OCH3-3),2.76(2H,m,CH2-α),2.39(2H,m,-CH2-β)；
from the above data, compound I was identified as 2, 6-bis (p-hydroxybenzyl) -5-hydroxy-3, 3' -dimethoxybibenzyl, having the following structure:
example 5 extraction and Structure identification of Compound J
(1) Extracting medicinal materials:
extracting dried rhizoma Bletillae coarse powder (3kg) with 95% ethanol under reflux for 3 times (30L × 3), each time for 2 hr;
(2) separation and purification of components:
firstly, concentrating and drying the ethanol extract (90L) under reduced pressure to obtain 510g of semisolid fluid extract;
dispersing the semi-solid fluid extract (500g) with water (5L), sequentially extracting with ethyl acetate and n-butanol (20L), combining ethyl acetate parts, and recovering the solvent under reduced pressure to obtain 160g of ethyl acetate extract;
③ separating the ethyl acetate extract (160g) by silica gel column chromatography, and purifying by using petroleum ether: gradient eluting with acetone, detecting the eluate by thin layer chromatography, and mixing similar components to obtain petroleum ether: eluent fr.18 when acetone is 20: 1;
the conditions for gradient elution were as follows:
processing the Fr.18 fraction (19g) by MCI, eluting by 40-100% ethanol water, collecting eluent containing a target compound according to thin-layer tracing, and recovering a solvent to obtain 15 components Fr.18-a-Fr.18-o;
fifthly, separating the Fr.18-m component (80% ethanol elution part, 0.9g) by a gel LH-20 chromatographic column, and sequentially eluting 1, 1 and 1 column volumes by using 60% methanol water as an eluent to respectively obtain six components Fr.18-m-1 to Fr.18-m-6;
subjecting Fr.18-m-2 fraction to gel LH-20 column chromatography (eluent dichloromethane-methanol 1:1), eluting for 4 column volumes, and subjecting to preparative thin layer chromatography (developing solvent dichloromethane-methanol 15:1, R)f0.68) and reverse phase high performance liquid chromatography with 55% methanol water to obtain compound J;
when the bibenzyl compounds in the rhizoma bletillae are developed by thin-layer chromatography, 10% sulfuric acid ethanol test solution is sprayed, and the rhizoma bletillae is developed for 3min at 105 ℃, so that yellow, orange-red to reddish spots are developed, and the rhizoma bletillae can be used for tracking detection of bibenzyl;
mass spectral data for compound J: ESI-MS M/z 561[ M-H ]]-；
Hydrogen spectrum data1H NMR(CD3COCD3,600MHz):7.04(2H,d,J＝8.4Hz,H-2″″,6″″),6.99(2H,d,J＝8.4Hz,H-2″,6″),6.92(2H,d,J＝8.4Hz,H-2″′,6″′),6.88(1H,d,J＝9.0Hz,H-5′),6.71(2H,d,J＝8.4Hz,H-3″″,5″″),6.69(1H,brs,H-2′),6.67(2H,d,J＝8.4Hz,H-3″,5″),6.66(2H,d,J＝8.4Hz,H-3″′,5″′),6.58(1H,s,H-4),6.52(1H,dd,J＝9.0,1.8Hz,H-6′),3.99(2H,s,CH2-7″),3.94(2H,s,CH2-7″′),3.80(2H,s,CH2-7″″),3.75(3H,s,OCH3),2.73(2H,m,CH2-α),2.33(2H,m,CH2-β)；
Carbon spectrum data:13C NMR(CD3COCD3,150MHz):157.8(C-3),156.2(C-4″″),156.0(C-4″),155.9(C-4″′),155.5(C-3′),155.4(C-5),142.3(C-1),142.3(C-1′),133.6(C-1″′),133.6(C-1″),133.1(C-1″″),131.1(C-5′),130.6(C-2″″,6″″),129.9(C-2″,6″),129.7(C-2″′),126.7(C-4′),120.2(C-6′),119.9(C-2),119.0(C-6),115.7(C-3″,5″；C-3″′,5″′；C-3″″,5″″),115.5(C-2′),97.8(C-4),55.7(C-OCH3),37.0(C-β),35.1(C-7″″),33.3(C-α),30.9(C-7″),30.8(C-7″′)；
from the above data, compound J is 2,4 ', 6-tris (p-hydroxybenzyl) -3', 5-dihydroxy-3-methoxybibenzyl, having the following structure:
the advantageous effects of the present invention will be specifically described below by way of test examples.
Example 6 use of Compounds F to J according to the invention for the treatment and/or prophylaxis of tumors
(1) Experimental materials:
(ii) cells
Lung cancer cell a549 was derived from ATCC company, usa.
Second experiment instrument
HWCL-1 constant temperature magnetic stirring bath (Zhengzhou great wall science and trade Co., Ltd.);
BP211D one tenth ten million electronic balance (Sartorius, switzerland);
model DZG-6050 vacuum drying oven (shanghai semen);
microplate reader (Thermo 3001, Thermo Fisher Scientific);
an electronic balance (model ESJ120-4, shenyang dragon electronic weighing instrument ltd);
ultra clean bench (MCV-B161F (T), SANYO, Japan);
microscope (Primo Vert, AxioCam ERc 5s, ZEISS);
CO2incubator (HH.CP-T, Shanghai Zixin scientific instruments Co., Ltd.).
(2) The experimental method comprises the following steps:
(ii) inoculation of cells
Digesting the cells in logarithmic growth phase with 0.25% pancreatin, preparing single cell suspension by using cell culture medium containing 10% FBS, counting by using cell counting plate, inoculating A549 tumor cells in good state into 96-well plate to make cell density be 4 × 103Each well was filled with 100. mu.L of cell suspension, incubated at 37 ℃ with 5% CO2Culturing in an incubator for 24 h.
② treatment with drugs
Starting from 20. mu.g/mL of each sample, the samples were diluted with a gradient of medium, 2-fold diluted, 5 drug concentrations were set, and duplicate wells were tested for each concentration. The drug is added into each hole with the concentration of 20, 10, 5, 2.5 and 1.25 mug/mL for 100 mug L, each concentration is provided with 3 multiple holes, and the steps are repeated for 3 times. The negative control group is a culture medium solution containing a proper amount of DMSO, and the blank control group is a culture medium and a solvent which do not contain cells. The 96-well plate was returned to the incubator and exposed for 72h at 37 ℃.
③ color development and IC50Is calculated by
After 72h, 20. mu.L of MTT solution (5g/L) was added to each well in the dark, incubation was continued for 4h, the supernatant was discarded, 150. mu.L of DMSO was added to each well, the mixture was placed on a shaker and shaken at a slow speed for 10min to dissolve formazan sufficiently, and then OD at a wavelength of 570nm was measured using a microplate reader.
IC by using improved Kouya method50＝log-1[Xm-i(∑P-0.5)]A calculation was performed wherein Xm: a designed logarithm of maximum concentration; i: log of each concentration multiple concentration; e, sigma P: sum of growth inhibition rates of each group; 0.5: an empirical constant. By repeating the experiment three times, 3 ICs were calculated50Values (two decimal places are reserved), and the average value is calculated; the test results are shown in Table 1.
TABLE 1 cytotoxic Activity IC of bibenzyl Compounds F-J of the present invention on A549 tumor cells50
| NO. | IC50(μg/mL) |
| Compound F | 5.13 |
| Compound G | 5.88 |
| Compound H | 4.83 |
| Compound I | 3.80 |
| Compound J | 2.01 |
The results show that the bibenzyl compound has good inhibition effect on tumor cells and can be used for preparing antitumor drugs.
In conclusion, the invention provides a new application of bibenzyl compounds, in particular to an application in preparing medicaments for treating and/or preventing tumors, which has good inhibition effect on tumor cells, particularly on lung cancer cells and IC thereof502-6 mug/mL; meanwhile, the preparation method of the compound is simple and convenient, mild in reaction conditions, convenient to operate and control, low in energy consumption, high in yield, low in cost and very suitable for industrial production.
Claims (10)
1. The application of the compound shown in the formula IV or the pharmaceutically acceptable salt, crystal form and solvate thereof in preparing the medicine for treating and/or preventing tumors:
wherein,
R1、R4、R6each independently selected from H or methyl;
R2、R3、R5、R7each independently selected from H or
2. Use according to claim 1, characterized in that: the tumor is cancer; further, the cancer is lung cancer.
3. A process for preparing compound F, characterized in that: it comprises the following steps:
a. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
b. taking the ethanol extract, and concentrating to obtain a fluid extract;
c. dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, mixing ethyl acetate parts, and recovering solvent to obtain ethyl acetate extract;
d. taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially carrying out gradient elution by using petroleum ether-acetone as an eluent at a ratio of 100:1, at a ratio of 50:1 and at a ratio of 30:1 to obtain petroleum ether: eluent fr.16 when acetone is 30: 1;
e. fr.16 is loaded on a reversed-phase polystyrene resin column, gradient elution is carried out by using ethanol-water of 10:90, 30:70, 50:50, 70:30 and 90:10 as eluent in sequence, and Fr.16-h of the ethanol-water of 90:10 is obtained;
f. fr.16-h is subjected to medium-pressure liquid chromatography, and gradient elution is sequentially carried out by using methanol-water of 20:80, 30:70 and 50:50 as eluent, so as to obtain Fr.16-h-3 when the methanol-water of 50: 50;
g. fr.16-h-3 was chromatographed on silica gel sequentially with dichloromethane-methanol ═ 1: 0. 100, and (2) a step of: 1 as eluent, eluting 4 column volumes respectively, collecting dichloromethane-methanol as 100: fr.16-h-3b at 1;
h. fr.16-h-3 was subjected to gel column chromatography with methanol-water 70:30 as the washRemoving solvent, eluting 4 column volumes, performing preparative thin layer chromatography with dichloromethane-methanol (30: 1) as developing agent, and collecting eluate, wherein R isfCompound F was isolated as 0.58.
4. The process for the preparation of compound F according to claim 3, characterized in that:
in the step d, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step e, the amount of Fr.16 is 15-20 g, and the gradient elution conditions are as follows:
in the step f, the amount of Fr.16-h is 4-6 g, and the gradient elution conditions are as follows:
5. a process for preparing compound G or compound H, characterized in that: it comprises the following steps:
i. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
ii. Taking the ethanol extract, and concentrating to obtain a fluid extract;
iii, dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, combining ethyl acetate parts, and recovering the solvent to obtain an ethyl acetate extract;
iv, taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially mixing petroleum ether: gradient elution is carried out by using acetone as an eluent at the ratio of 100:1, 50:1 and 30:1, and petroleum ether is obtained: eluent fr.16 when acetone is 30: 1;
v, fr.16 on a reversed-phase polystyrene type resin column, sequentially mixing ethanol: gradient elution is carried out by using water as eluent, wherein the water is 10:90, 30:70, 50:50, 70:30, 90:10 and 100:0, and ethanol is obtained: fr.16-j when water is 100: 0;
vi, subjecting the Fr.16-j to a sephadex chromatographic column, and sequentially eluting by using dichloromethane-methanol (1: 1) as an eluent, wherein the eluent has the volume of 0.8 time of the column volume and 2.2 times of the column volume to obtain Fr.16-j-1 and Fr.16-j-2 respectively;
vii and fr.16-j-2, silica gel column chromatography, sequentially eluting with dichloromethane-methanol ═ 1: 0. 200: eluting with eluent 1, eluting 3 column volumes respectively, and collecting dichloromethane-methanol (200): fr.16-j-2b at 1;
viii, and Fr.16-j-2b are subjected to sephadex chromatographic column, and petroleum ether-dichloromethane-methanol 2:2:1 is used as eluent, and 1 column volume are sequentially eluted to obtain Fr.16-j-2bA and Fr.16-j-2 bB;
fr.16-j-2bA by preparative thin layer chromatography with dichloromethane-acetone 20:1 as developing solvent, Rf(iv) isolating as 0.76 to give compound G;
fr.16-j-2bB by preparative thin layer chromatography using dichloromethane-acetone 20:1 as developing solvent, Rf0.80, isolated to give compound H.
6. The process according to claim 5 for the preparation of compound G or compound H, characterized in that:
in the step iv, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step v, the amount of Fr.16 is 15-20 g, and gradient elution conditions are as follows:
7. a process for preparing compound I, characterized in that: it comprises the following steps:
A. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
B. taking the ethanol extract, and concentrating to obtain a fluid extract;
C. dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, mixing ethyl acetate parts, and recovering solvent to obtain ethyl acetate extract;
D. taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially mixing petroleum ether: gradient elution is carried out by using acetone as eluent, wherein the acetone is 100:1, 50:1, 30:1 and 20:1, and petroleum ether is obtained: eluent fr.18 when acetone is 20: 1;
E. fr.18 on a reversed-phase polystyrene type resin column, sequentially eluting with ethanol: gradient elution is carried out by using water as eluent and water as eluent, wherein the water is 40:60, 50:50, 55:45, 60:40, 70:30, 80:20 and 95:5 are carried out to obtain ethanol: fr.18-n at 95: 5;
F. sequentially eluting with dichloromethane-methanol (1: 1) as eluent by Fr.18-n sephadex chromatographic column to obtain Fr.18-n-1, Fr.18-n-2 and Fr.18-n-3 respectively;
fr.18-n-3 was chromatographed on Sephadex column with methanol-water 70:30 as eluent for 3 column volumes, the main compound was chromatographed on preparative thin layer chromatography with dichloromethane-methanol 20:1 as developing solvent, RfThen purified by reverse phase high performance liquid chromatography with methanol-water 65:35 as the mobile phase to give compound I.
8. The process for the preparation of compound I according to claim 7, characterized in that:
in the step D, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step E, the amount of Fr.18 is 18-20 g, and the gradient elution conditions are as follows:
9. a process for preparing compound J, characterized in that: it comprises the following steps:
I. extracting rhizoma Bletillae with ethanol to obtain ethanol extractive solution;
II. Taking the ethanol extract, and concentrating to obtain a fluid extract;
III, dispersing the fluid extract with water, sequentially extracting with ethyl acetate and n-butanol, combining ethyl acetate parts, and recovering the solvent to obtain an ethyl acetate extract;
IV, taking ethyl acetate extract, adopting silica gel column chromatography, and sequentially mixing petroleum ether: gradient elution is carried out by using acetone as eluent, wherein the acetone is 100:1, 50:1, 30:1 and 20:1, and petroleum ether is obtained: eluent fr.18 when acetone is 20: 1;
v, Fr.18 on a reversed-phase polystyrene resin column, sequentially adding ethanol: gradient elution is carried out by using water as eluent and water as eluent, wherein the water is 40:60, 50:50, 55:45, 60:40, 70:30 and 80:20, and ethanol is obtained: fr.18-m at 80: 20;
VI and Fr.18-m are put on a sephadex chromatographic column, and are sequentially eluted by 1 time of column volume and 1 time of column volume by using methanol-water (60: 40) as an eluent to respectively obtain Fr.18-m-1 and Fr.18-m-2;
fr.18-m-2 was passed through a Sephadex column chromatography toEluting with dichloromethane-methanol (1: 1) as eluent for 4 column volumes, performing preparative thin layer chromatography with dichloromethane-methanol (15: 1) as developing agent, and RfThen purified by reverse phase high performance liquid chromatography with methanol-water 55:45 as the mobile phase to give compound J.
10. The process for preparing compound J according to claim 9, characterized in that:
in the step IV, the amount of the ethyl acetate extract is 155-165 g, and the conditions of gradient elution are as follows:
in the step V, the amount of Fr.18 is 18-20 g, and the gradient elution conditions are as follows:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610719859.7A CN106361731B (en) | 2016-08-24 | 2016-08-24 | Bibenzyl compound, preparation method thereof and application thereof in preparing antitumor drugs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610719859.7A CN106361731B (en) | 2016-08-24 | 2016-08-24 | Bibenzyl compound, preparation method thereof and application thereof in preparing antitumor drugs |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106361731A true CN106361731A (en) | 2017-02-01 |
| CN106361731B CN106361731B (en) | 2020-02-04 |
Family
ID=57879539
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610719859.7A Expired - Fee Related CN106361731B (en) | 2016-08-24 | 2016-08-24 | Bibenzyl compound, preparation method thereof and application thereof in preparing antitumor drugs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106361731B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110357847A (en) * | 2019-07-22 | 2019-10-22 | 四川大学 | Isoflavane compound and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103933350A (en) * | 2012-12-31 | 2014-07-23 | 成都中医药大学 | Use of rhizoma bletillae ethyl acetate extract |
| CN105001064A (en) * | 2015-07-10 | 2015-10-28 | 浙江中医药大学 | Bletilla striata fibrous root blestriarene A and preparation method and application thereof |
-
2016
- 2016-08-24 CN CN201610719859.7A patent/CN106361731B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103933350A (en) * | 2012-12-31 | 2014-07-23 | 成都中医药大学 | Use of rhizoma bletillae ethyl acetate extract |
| CN105001064A (en) * | 2015-07-10 | 2015-10-28 | 浙江中医药大学 | Bletilla striata fibrous root blestriarene A and preparation method and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| HIROSHI MORITA 等: "Antimitotic activity and reversal of breast cancer resistance protein-mediated drug resistance by stilbenoids from Bletilla striata", 《BIOORGANIC & MEDICINAL CHEMISTRY LETTERS》 * |
| JIA-QUAN FENG, 等: ""Novel Bibenzyl Derivatives from the Tubers of Bletilla striata"", 《HELVETICA CHIMICA ACTA》 * |
| ZCAPLUS: "847670-85-3，87530-28-7", 《STN》 * |
| 沈凯凯，等: ""石斛中联苄类化合物抗血管新生活性研究"", 《时珍国医国药》 * |
| 韩广轩 等: ""中药白及中一新的联苄化合物"", 《药学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110357847A (en) * | 2019-07-22 | 2019-10-22 | 四川大学 | Isoflavane compound and preparation method and application thereof |
| CN110357847B (en) * | 2019-07-22 | 2022-05-13 | 四川大学 | Isoflavane compound and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106361731B (en) | 2020-02-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110272342B (en) | Naphthoic acid compound in purslane and extraction and separation method and application thereof | |
| CA3081627A1 (en) | Method for separating eight components in chinese traditional medicine composition | |
| CN110343116A (en) | A kind of Flos Chrysanthemi Indici extract and preparation method thereof and the application in preparation treatment medicine for nasopharyngeal | |
| CN115160337A (en) | 1 alpha-alkyl daphnane diterpenoid compounds, and preparation method and application thereof | |
| CN106361731B (en) | Bibenzyl compound, preparation method thereof and application thereof in preparing antitumor drugs | |
| CN110357847B (en) | Isoflavane compound and preparation method and application thereof | |
| CN109879921B (en) | Compound separated from rhizoma anemarrhenae and having antitumor activity and preparation method thereof | |
| CN106431852B (en) | Bibenzyl compound and preparation method thereof and purposes in the preparation of antitumor drugs | |
| CN106495998B (en) | A kind of bibenzyl ether compound and preparation method thereof and purposes | |
| CN111548327A (en) | Carbon-reduced kaurane diterpene, preparation method thereof and application thereof in preparation of antitumor drugs | |
| CN106349032B (en) | Bibenzyl compound and preparation method thereof and purposes in the preparation of antitumor drugs | |
| CN106349031B (en) | Bibenzyl compound and preparation method thereof and purposes in the preparation of antitumor drugs | |
| CN109824685B (en) | Compound oleracene G in purslane, extraction and separation method and application thereof | |
| CN103183597B (en) | Preparing method of diaryl neptanone compound having antineoplastic activity, and application | |
| CN106397140B (en) | A kind of poly bibenzyl derivative and preparation method thereof and purposes | |
| CN110563563B (en) | Chalcone compound and preparation method and application thereof | |
| Feu et al. | Glycosylated narciclasine alkaloid in Hippeastrum puniceum (Lam.) Kuntze | |
| CN111377933B (en) | Alkaloid compound extracted from orychophragmus violaceus seeds as well as extraction method and application thereof | |
| CN109879926B (en) | Triterpene glycoside compounds in Glechomae herba and extraction and separation method thereof | |
| CN106822088B (en) | Application of diene iridoid compound in preparation of anti-cancer drugs | |
| CN110272464B (en) | Compound, preparation method thereof and application thereof in preparing antitumor drugs | |
| CN101412726B (en) | Novel heterocyclic compounds | |
| CN112300185B (en) | Alkaloid compound with reduced hepatotoxicity, and preparation method and application thereof | |
| CN110526809B (en) | Chalcone compound and preparation method and application thereof | |
| CN111171100B (en) | Limonin compound with anti-tumor activity and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20200204 Termination date: 20210824 |