CN113354609B - 一种异戊烯基取代香豆素类化合物及其制备方法和应用 - Google Patents
一种异戊烯基取代香豆素类化合物及其制备方法和应用 Download PDFInfo
- Publication number
- CN113354609B CN113354609B CN202110745925.9A CN202110745925A CN113354609B CN 113354609 B CN113354609 B CN 113354609B CN 202110745925 A CN202110745925 A CN 202110745925A CN 113354609 B CN113354609 B CN 113354609B
- Authority
- CN
- China
- Prior art keywords
- methanol
- petroleum ether
- compound
- preparation
- water
- 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
- -1 Isopentenyl Chemical group 0.000 title claims abstract description 30
- 238000002360 preparation method Methods 0.000 title claims abstract description 15
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N benzo-alpha-pyrone Natural products C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 title claims abstract description 13
- 229960000956 coumarin Drugs 0.000 title claims abstract description 13
- 235000001671 coumarin Nutrition 0.000 title claims abstract description 13
- 150000001875 compounds Chemical class 0.000 claims abstract description 30
- 239000000126 substance Substances 0.000 claims abstract description 17
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 claims abstract description 15
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 claims abstract description 15
- 238000000926 separation method Methods 0.000 claims abstract description 6
- 238000000746 purification Methods 0.000 claims abstract description 5
- 239000003208 petroleum Substances 0.000 claims description 35
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 claims description 28
- 239000012259 ether extract Substances 0.000 claims description 16
- 239000002246 antineoplastic agent Substances 0.000 claims description 14
- 229940041181 antineoplastic drug Drugs 0.000 claims description 14
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 claims description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 12
- 238000004440 column chromatography Methods 0.000 claims description 12
- 210000004881 tumor cell Anatomy 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- 206010028980 Neoplasm Diseases 0.000 claims description 5
- 239000000469 ethanolic extract Substances 0.000 claims description 5
- 239000000178 monomer Substances 0.000 claims description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 239000000706 filtrate Substances 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 239000000049 pigment Substances 0.000 claims description 4
- 238000002953 preparative HPLC Methods 0.000 claims description 4
- 238000010298 pulverizing process Methods 0.000 claims description 4
- 239000011347 resin Substances 0.000 claims description 4
- 229920005989 resin Polymers 0.000 claims description 4
- 238000010898 silica gel chromatography Methods 0.000 claims description 4
- 239000000725 suspension Substances 0.000 claims description 4
- 239000000284 extract Substances 0.000 claims description 3
- 230000008685 targeting Effects 0.000 claims description 3
- 238000010828 elution Methods 0.000 claims description 2
- 125000000332 coumarinyl group Chemical class O1C(=O)C(=CC2=CC=CC=C12)* 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract description 22
- 230000000259 anti-tumor effect Effects 0.000 abstract description 15
- 229940079593 drug Drugs 0.000 abstract description 13
- 230000000694 effects Effects 0.000 abstract description 12
- 235000012907 honey Nutrition 0.000 abstract description 7
- 239000013641 positive control Substances 0.000 abstract description 5
- 238000011156 evaluation Methods 0.000 abstract description 4
- 230000002401 inhibitory effect Effects 0.000 abstract description 4
- 238000006243 chemical reaction Methods 0.000 abstract description 3
- 230000005918 in vitro anti-tumor Effects 0.000 abstract 1
- 241000218232 Artocarpus Species 0.000 description 16
- 230000005764 inhibitory process Effects 0.000 description 14
- 235000013399 edible fruits Nutrition 0.000 description 13
- 241000196324 Embryophyta Species 0.000 description 10
- 239000000243 solution Substances 0.000 description 8
- 241000282414 Homo sapiens Species 0.000 description 7
- 229930014626 natural product Natural products 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 6
- 239000003480 eluent Substances 0.000 description 6
- 229940121358 tyrosine kinase inhibitor Drugs 0.000 description 6
- 239000005483 tyrosine kinase inhibitor Substances 0.000 description 6
- 244000025352 Artocarpus heterophyllus Species 0.000 description 5
- 229930003935 flavonoid Natural products 0.000 description 5
- 235000017173 flavonoids Nutrition 0.000 description 5
- HTSGKJQDMSTCGS-UHFFFAOYSA-N 1,4-bis(4-chlorophenyl)-2-(4-methylphenyl)sulfonylbutane-1,4-dione Chemical compound C1=CC(C)=CC=C1S(=O)(=O)C(C(=O)C=1C=CC(Cl)=CC=1)CC(=O)C1=CC=C(Cl)C=C1 HTSGKJQDMSTCGS-UHFFFAOYSA-N 0.000 description 4
- HIXDQWDOVZUNNA-UHFFFAOYSA-N 2-(3,4-dimethoxyphenyl)-5-hydroxy-7-methoxychromen-4-one Chemical compound C=1C(OC)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(OC)C(OC)=C1 HIXDQWDOVZUNNA-UHFFFAOYSA-N 0.000 description 4
- 235000008725 Artocarpus heterophyllus Nutrition 0.000 description 4
- 150000002215 flavonoids Chemical class 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 229930008679 prenylflavonoid Natural products 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 150000004917 tyrosine kinase inhibitor derivatives Chemical class 0.000 description 4
- 244000238680 Artocarpus lakoocha Species 0.000 description 3
- 235000003328 Artocarpus lakoocha Nutrition 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 241000218231 Moraceae Species 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 230000000144 pharmacologic effect Effects 0.000 description 3
- PJANXHGTPQOBST-UHFFFAOYSA-N stilbene Chemical class C=1C=CC=CC=1C=CC1=CC=CC=C1 PJANXHGTPQOBST-UHFFFAOYSA-N 0.000 description 3
- 150000003648 triterpenes Chemical class 0.000 description 3
- PQNTWKDHNSWVPU-UHFFFAOYSA-N 9,19-cyclo-9β-lanost-24-en-3β-ol, acetate Chemical compound C1CC(OC(C)=O)C(C)(C)C2C31CC13CCC3(C)C(C(CCC=C(C)C)C)CCC3(C)C1CC2 PQNTWKDHNSWVPU-UHFFFAOYSA-N 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 2
- 241001441970 Anserinae Species 0.000 description 2
- 240000004161 Artocarpus altilis Species 0.000 description 2
- 235000002672 Artocarpus altilis Nutrition 0.000 description 2
- 241001471167 Artocarpus lanceifolius Species 0.000 description 2
- 241001573881 Corolla Species 0.000 description 2
- 239000005517 L01XE01 - Imatinib Substances 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000006838 adverse reaction Effects 0.000 description 2
- 230000000843 anti-fungal effect Effects 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 230000000078 anti-malarial effect Effects 0.000 description 2
- 230000000840 anti-viral effect Effects 0.000 description 2
- 239000003430 antimalarial agent Substances 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 244000309466 calf Species 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 238000012790 confirmation Methods 0.000 description 2
- YNBJLDSWFGUFRT-UHFFFAOYSA-N cycloartenol Natural products CC(CCC=C(C)C)C1CCC2(C)C1(C)CCC34CC35CCC(O)C(C)(C)C5CCC24C YNBJLDSWFGUFRT-UHFFFAOYSA-N 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 229930003944 flavone Natural products 0.000 description 2
- 235000011949 flavones Nutrition 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000002114 high-resolution electrospray ionisation mass spectrometry Methods 0.000 description 2
- KTUFNOKKBVMGRW-UHFFFAOYSA-N imatinib Chemical compound C1CN(C)CCN1CC1=CC=C(C(=O)NC=2C=C(NC=3N=C(C=CN=3)C=3C=NC=CC=3)C(C)=CC=2)C=C1 KTUFNOKKBVMGRW-UHFFFAOYSA-N 0.000 description 2
- 229960002411 imatinib Drugs 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 239000000401 methanolic extract Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 150000007951 prenylflavonoids Chemical class 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 238000004611 spectroscopical analysis Methods 0.000 description 2
- 210000000952 spleen Anatomy 0.000 description 2
- 235000021286 stilbenes Nutrition 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 230000009897 systematic effect Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- XZEUYTKSAYNYPK-UHFFFAOYSA-N 3beta-29-Norcycloart-24-en-3-ol Natural products C1CC2(C)C(C(CCC=C(C)C)C)CCC2(C)C2CCC3C(C)C(O)CCC33C21C3 XZEUYTKSAYNYPK-UHFFFAOYSA-N 0.000 description 1
- YRNWIFYIFSBPAU-UHFFFAOYSA-N 4-[4-(dimethylamino)phenyl]-n,n-dimethylaniline Chemical compound C1=CC(N(C)C)=CC=C1C1=CC=C(N(C)C)C=C1 YRNWIFYIFSBPAU-UHFFFAOYSA-N 0.000 description 1
- 208000007848 Alcoholism Diseases 0.000 description 1
- AAXZLRPBJQFZQA-UHFFFAOYSA-N Artocarpin Natural products CC(C)=CCC=1C(=O)C=2C(O)=C(CC=C(C)C)C(OC)=CC=2OC=1C1=CC=C(O)C=C1O AAXZLRPBJQFZQA-UHFFFAOYSA-N 0.000 description 1
- 235000008727 Artocarpus polyphema Nutrition 0.000 description 1
- 241000209128 Bambusa Species 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 108010004032 Bromelains Proteins 0.000 description 1
- DQFBYFPFKXHELB-UHFFFAOYSA-N Chalcone Natural products C=1C=CC=CC=1C(=O)C=CC1=CC=CC=C1 DQFBYFPFKXHELB-UHFFFAOYSA-N 0.000 description 1
- 108010066551 Cholestenone 5 alpha-Reductase Proteins 0.000 description 1
- 206010053567 Coagulopathies Diseases 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- RRTBTJPVUGMUNR-UHFFFAOYSA-N Cycloartanol Natural products C12CCC(C(C(O)CC3)(C)C)C3C2(CC)CCC2(C)C1(C)CCC2C(C)CCCC(C)C RRTBTJPVUGMUNR-UHFFFAOYSA-N 0.000 description 1
- 101100136092 Drosophila melanogaster peng gene Proteins 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 206010060710 Galactostasis Diseases 0.000 description 1
- 240000005323 Hoya carnosa Species 0.000 description 1
- 206010020772 Hypertension Diseases 0.000 description 1
- 238000004566 IR spectroscopy Methods 0.000 description 1
- HVXLSFNCWWWDPA-UHFFFAOYSA-N Isocycloartenol Natural products C1CC(O)C(C)(C)C2C31CC13CCC3(C)C(C(CCCC(C)=C)C)CCC3(C)C1CC2 HVXLSFNCWWWDPA-UHFFFAOYSA-N 0.000 description 1
- 239000005411 L01XE02 - Gefitinib Substances 0.000 description 1
- 239000005551 L01XE03 - Erlotinib Substances 0.000 description 1
- 208000019395 Lactation disease Diseases 0.000 description 1
- 229930189806 Lakoochin Natural products 0.000 description 1
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 1
- 235000010654 Melissa officinalis Nutrition 0.000 description 1
- 108010090306 Member 2 Subfamily G ATP Binding Cassette Transporter Proteins 0.000 description 1
- 241000730022 Millettia erythrocalyx Species 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- HXQRIQXPGMPSRW-UHZRDUGNSA-N Pollinastanol Natural products O[C@@H]1C[C@H]2[C@@]3([C@]4([C@H]([C@@]5(C)[C@@](C)([C@H]([C@H](CCCC(C)C)C)CC5)CC4)CC2)C3)CC1 HXQRIQXPGMPSRW-UHZRDUGNSA-N 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 241000015864 Protobothrops flavoviridis Species 0.000 description 1
- 208000001431 Psychomotor Agitation Diseases 0.000 description 1
- 206010038743 Restlessness Diseases 0.000 description 1
- 244000300264 Spinacia oleracea Species 0.000 description 1
- 235000009337 Spinacia oleracea Nutrition 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- PJANXHGTPQOBST-VAWYXSNFSA-N Stilbene Natural products C=1C=CC=CC=1/C=C/C1=CC=CC=C1 PJANXHGTPQOBST-VAWYXSNFSA-N 0.000 description 1
- 206010042576 Suppressed lactation Diseases 0.000 description 1
- 241000973887 Takayama Species 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- LUKBXSAWLPMMSZ-OWOJBTEDSA-N Trans-resveratrol Chemical class C1=CC(O)=CC=C1\C=C\C1=CC(O)=CC(O)=C1 LUKBXSAWLPMMSZ-OWOJBTEDSA-N 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 102000003425 Tyrosinase Human genes 0.000 description 1
- 108060008724 Tyrosinase Proteins 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 201000007930 alcohol dependence Diseases 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000001355 anti-mycobacterial effect Effects 0.000 description 1
- 230000000702 anti-platelet effect Effects 0.000 description 1
- 230000001028 anti-proliverative effect Effects 0.000 description 1
- 230000002365 anti-tubercular Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 239000003926 antimycobacterial agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- KRGDFVQWQJIMEK-RMKNXTFCSA-N artocarpin Chemical compound CC(C)=CCC=1C(=O)C=2C(O)=C(\C=C\C(C)C)C(OC)=CC=2OC=1C1=CC=C(O)C=C1O KRGDFVQWQJIMEK-RMKNXTFCSA-N 0.000 description 1
- 108010034967 artocarpin lectin Proteins 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 235000019835 bromelain Nutrition 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 235000005513 chalcones Nutrition 0.000 description 1
- NAJCQAAOHKVCES-ZZOQNIIDSA-N chembl226084 Chemical compound C1CC(=O)C(C)(C)[C@H]2[C@@]31C[C@@]13CC[C@]3(C)[C@@H]([C@@H](CCC=C(C)C)C)CC[C@@]3(C)[C@@H]1CC2 NAJCQAAOHKVCES-ZZOQNIIDSA-N 0.000 description 1
- OAIVIYSBZFEOIU-UHFFFAOYSA-N chloroform;propan-2-one Chemical compound CC(C)=O.ClC(Cl)Cl OAIVIYSBZFEOIU-UHFFFAOYSA-N 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 230000035602 clotting Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 150000001893 coumarin derivatives Chemical class 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- ONQRKEUAIJMULO-YBXTVTTCSA-N cycloartenol Chemical compound CC(C)([C@@H](O)CC1)[C@H]2[C@@]31C[C@@]13CC[C@]3(C)[C@@H]([C@@H](CCC=C(C)C)C)CC[C@@]3(C)[C@@H]1CC2 ONQRKEUAIJMULO-YBXTVTTCSA-N 0.000 description 1
- FODTZLFLDFKIQH-UHFFFAOYSA-N cycloartenol trans-ferulate Natural products C1=C(O)C(OC)=CC(C=CC(=O)OC2C(C3CCC4C5(C)CCC(C5(C)CCC54CC53CC2)C(C)CCC=C(C)C)(C)C)=C1 FODTZLFLDFKIQH-UHFFFAOYSA-N 0.000 description 1
- CDTIDJDMSJTMJX-UHFFFAOYSA-N cycloartenone Natural products C12CCC(C(C(=O)CC3)(C)C)C3C2(CC)CCC2(C)C1(C)CCC2C(C)CCC=C(C)C CDTIDJDMSJTMJX-UHFFFAOYSA-N 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 208000001848 dysentery Diseases 0.000 description 1
- 235000018927 edible plant Nutrition 0.000 description 1
- 230000001516 effect on protein Effects 0.000 description 1
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 1
- 229960001433 erlotinib Drugs 0.000 description 1
- AAKJLRGGTJKAMG-UHFFFAOYSA-N erlotinib Chemical compound C=12C=C(OCCOC)C(OCCOC)=CC2=NC=NC=1NC1=CC=CC(C#C)=C1 AAKJLRGGTJKAMG-UHFFFAOYSA-N 0.000 description 1
- HWJHWSBFPPPIPD-UHFFFAOYSA-N ethoxyethane;propan-2-one Chemical compound CC(C)=O.CCOCC HWJHWSBFPPPIPD-UHFFFAOYSA-N 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 150000002213 flavones Chemical class 0.000 description 1
- 229960002584 gefitinib Drugs 0.000 description 1
- XGALLCVXEZPNRQ-UHFFFAOYSA-N gefitinib Chemical compound C=12C=C(OCCCN3CCOCC3)C(OC)=CC2=NC=NC=1NC1=CC=C(F)C(Cl)=C1 XGALLCVXEZPNRQ-UHFFFAOYSA-N 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 230000002443 hepatoprotective effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 201000005202 lung cancer Diseases 0.000 description 1
- 208000020816 lung neoplasm Diseases 0.000 description 1
- 201000004792 malaria Diseases 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 210000002418 meninge Anatomy 0.000 description 1
- 230000036457 multidrug resistance Effects 0.000 description 1
- 208000025113 myeloid leukemia Diseases 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 235000017807 phytochemicals Nutrition 0.000 description 1
- 229930000223 plant secondary metabolite Natural products 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 102000027426 receptor tyrosine kinases Human genes 0.000 description 1
- 108091008598 receptor tyrosine kinases Proteins 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000012306 spectroscopic technique Methods 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 150000001629 stilbenes Chemical class 0.000 description 1
- 239000012089 stop solution Substances 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- DQFBYFPFKXHELB-VAWYXSNFSA-N trans-chalcone Chemical compound C=1C=CC=CC=1C(=O)\C=C\C1=CC=CC=C1 DQFBYFPFKXHELB-VAWYXSNFSA-N 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
- 150000007964 xanthones Chemical class 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/06—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2
- C07D311/08—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring
- C07D311/16—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted in position 7
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/60—Moraceae (Mulberry family), e.g. breadfruit or fig
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Alternative & Traditional Medicine (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及天然药物领域，公开一种来源于波罗蜜的具有新颖化学结构的异戊烯基取代香豆素类化合物及其制备方法和在抗肿瘤药物中的应用，所述化合物artoheteronin经多种体外抗肿瘤活性评价结果表明：该化合物具有显著的抗肿瘤活性，具有与阳性对照药相当的抑制蛋白酪氨酸激酶的活性，可以进一步开发成以蛋白酪氨酸激酶为靶标的抗肿瘤药物，可在抗肿瘤药物中应用，分离纯化工艺简单，反应条件温和，具有现实意义。
Description
技术领域
本发明属于天然药物领域，具体涉及一种来源于菠罗蜜的具有新颖化学结构的异戊烯基取代香豆素类化合物的制备方法及其在抗肿瘤药物中的应用。
背景技术
恶性肿瘤是一种多发病，死亡率较高，目前正严重威胁着人类的生命与健康，且正逐渐成为人类生命和健康的头号杀手。现代医学对恶性肿瘤的治疗主要包括手术治疗、化学药物治疗、放射治疗和生物治疗，以化学药物治疗为主，即使采用了手术治疗的方式，往往也需要在手术后常规采用化学药物进行辅助治疗。随着生命科学的飞速发展，恶性肿瘤的发生机制的不断被阐明，新的抗肿瘤作用靶点不断被发现，靶向抑制肿瘤信号转导成为新型抗肿瘤药物开发的重要方向之一。分子靶向抗肿瘤药物能够针对肿瘤细胞生长和增殖过程中的特异性靶点选择性地杀伤肿瘤细胞而不损伤正常组织，给肿瘤治疗领域带来了革命性的进展。在各种分子靶点中，蛋白酪氨酸激酶(Protein Tyrosine Kinase，PTK)是目前研究最多且效果最明显的抗肿瘤药物靶点之一，已成为抗肿瘤靶向治疗药物的研究重点和热点。目前临床上使用的小分子靶向抗肿瘤药物主要为酪氨酸激酶抑制剂类抗肿瘤药物，包括单靶点酪氨酸激酶抑制剂和多靶点酪氨酸激酶抑制剂。临床实践同样表明，单靶点酪氨酸激酶抑制剂类抗肿瘤药物，如厄洛替尼和吉非替尼，尽管选择性强、毒副作用小，但在使用过程中易产生耐药性，无法彻底杀灭肿瘤细胞，而联合用药又会带来严重的不良反应，影响各自的药动学特性。多靶点酪氨酸激酶抑制剂具有减少药物使用种类，可从多方面发挥效用，对抗耐药性，避免产生药物相互作用，减少不良反应等优点。许多多靶点酪氨酸激酶抑制剂类抗肿瘤药物因其选择性高、疗效好、毒性小的临床特点，已逐渐成为某些肿瘤治疗的一线用药。
桑科(Moraceae)波罗蜜属(Artocarpus)植物全世界约有50种，分布于热带亚洲，自斯里兰卡、印度、巴基斯坦、孟加拉国、喜马拉雅山区各国、缅甸、泰国、中南半岛各国、马来西亚、印度尼西亚、巴布亚新几内亚至所罗门群岛等地。分布于我国的波罗蜜属植物约有15种以及2亚种，主要分布于海南、广东、广西、云南、贵州，福建以及台湾等省区[中国科学院中国植物志编委会.中国植物志.第23(1)卷.科学出版社.北京:1998,pp 40-55.]。在民间，波罗蜜属植物作药用植物已有很长历史，常用于肝硬化、高血压、风湿病、疟疾、痢疾以及肺结核等疾病的治疗；现代药理学研究表明该属植物中的化学成分具有广泛的生物活性如抗肿瘤、抗炎、抗疟、抗结核、抗真菌、抗病毒、抗血小板凝结以及抗氧化等活性[Fang,S.C.；Hsu,C.L.；Yen,G.C.Anti-inflammatory effects of phenolic compoundsisolated from the fruits of Artocarpus heterophyllus.Journal ofAgriculturaland Food Chemistry 2008,56,4463-4468.；Zheng,Z.P.；Xu,Y.；Qin,C.；Zhang,S.；Gu,X.H.；Lin,Y.Y.；Xie,G.B.；Wang,M.F.；Chen,J.Characterization of antiproliferativeactivity constituents from Artocarpus heterophyllus.Journal of Agriculturaland Food Chemistry 2014,62,5519-5527.；Sun,G.C.；Zheng,Z.P.；Lee,M.H.；Xu,Y.J.；Kang,S.；Dong,Z.G.；Wang,M.F.；Gu,Z.N.；Li,H.T.；Chen,W.Chemoprevention ofcolorectal cancerby artocarpin,a aietary phytochemical from Artocarpusheterophyllus.Journal of Agricultural and Food Chemistry 2017,65,3474-3480.；Puntumchai,A.；Kittakoop,P.；Rajviroongit,S.；Vimuttipong,S.；Likhitwitayawuid,K.；Thebtaranonth,Y.Lakoochins A and B,New antimycobacterial stilbenederivatives from Artocarpus lakoocha.Journal of Natural Products 2004,67,485-486.；Boonlaksiri,C.；Oonanant,W.；Kongsaeree,P.；Kittakoop,P.；Tanticharoen,M.；Thebtaranonth,Y.An antimalarial stilbene from Artocarpusinteger.Phytochemistry 2000,54,415-417.；Likhitwitayawuid,K.；Sritularak,B.；Benchanak,K.；Lipipun,V.；Mathew,J.；Schinazi,R.F.Phenolics with antiviralactivity from Millettia erythrocalyx and Artocarpus lakoocha.Natural ProductResearch 2005,19,177-182.；Jayasinghe,L.；Balasooriya,B.S.；Padmini,W.C.；Hara,N.；Fujimoto,Y.Geranyl chalcone derivatives with antifungal and cadicalscavenging properties from the leaves of Artocarpus nobilis.Phytochemistry2004,65,1287-1290.；Lin,K.W.；Liu,C.H.；Tu,H.Y.；Ko,H.H.；Wei,B.L.Antioxidantprenylflavonoids from Artocarpus communis and Artocarpus elasticus.FoodChemistry 2009,115,558-562.；Weng,J.R.；Chan,S.C.；Lu,Y.H.；Lin,H.C.；Ko,H.H.；Lin,C.N.Antiplatelet prenylflavonoids from Artocarpus communis.Phytochemistry2006,67,824-829.]。正因为如此，该属植物一直备受植物化学和药理学界科研工作者的青睐。自1971年起，对该属植物中的化学成分及其药理活性的研究一直是天然产物化学的热点之一。目前国内外学者从该属植物中共分离得到220余个化合物，包括黄酮类、芪类、甾体类和三萜类等多种类型化合物，其中结构多样性异常丰富的多异戊烯基取代的黄酮类化合物为其主要化合物类型[Chakravarti,R.N.；Mahato,S.B.；Banerjee,S.K.Triterpenes ofthe stem-bark of Artocarpus chaplasha.Phytochemistry 1971,10,1351-1354.；Pavanassaviam,G.；Sultabawa,M.S.Cycloartenyl acetate,cycloartenol,andcycloartenone in the bark of Artocarpus species.Phytochemistry 1973,12,2725-2726.；Altman,L.J.；Zito,S.W.Sterols and triterpenes from the fruit ofArtocarpus altilis.Phytochemistry 1976,15,829-830.；Wongkham,S.；Wongkham,C.；Boonsiri,P.；Simasathiansophon,S.；Trisonthi,C.；Atisook,K.Isoelectins fromseeds ofArtocarpus lakoocha.Phytochemistry 1995,40,1331-1334.；Lin,C.N.；Lu,C.M.；Huang,P.L.Flavonoids fromArtocarpus heterophyllus.Phytochemistry 1995,39,1447-1451.；Shimizu,K.；Kondo,R.；Sakai,K.；Buabarn,S.；Dilokkunanant,U.Ageranylated chalcone with 5α-reductase inhibitory properties from Artocarpusincisus.Phytochemistry 2000,54,737-739.；Syah,Y.M.；Achmad,S.A.；Ghisalberti,E,L.；Hakim,E.H.；Makmur,L.；Mujahidin,D.Artoindonesianins G-I,three newisoprenylated flavones from Artocarpus lanceifolius.Fitoterapia 2001,72,765-773.；Hakim,E.H.；Ulinnuha,U.Z.；Offen,P.；Syah,Y.M.；Killmer,L.；Ghisalberti,E.L.Artoindonesianins N and O,new prenylated stilbene and prenylatedarylbenzofuran derivatives from Artocarpus gomezianus.Fitoterapia 2002,73,597-603.；Hakim,E.H.；Asnizar；Y.；Kitajima,M.；Takayama,H.Artoindonesianin P,anew prenylated flavone with cytotoxic activity from Artocarpuslanceifolius.Fitoterapia 2002,73,668-673.；Shen,H.；Hou,A.J.Prenylated 2-arylbenzofurans from Artocarpus petelotii.Natural Product Research 2008,22,1451-1456.；Jayasinghe,U.B.；Samarakoon,T.B.；Kumarihamy,B.M.；Hara,N.；Fujimoto,Y.Four new prenylated flavonoids and xanthones from the root barkofArtocarpus nobilis.Fitoterapia 2008,79,37-41.；Versiani,M.A.；Diyabalanage,T.；Ratnayake,R.；Henrich,C.J.；Bates,S.E.；McMahon,J.B.；Gustafson,K.R.Flavonoidsfrom eight tropical plant species that inhibit the multidrug resistancetransporter ABCG2.Journal of Natural Products 2011,74,262-266.；Nguyen,N.T.；Nguyen,M.K.；Diyabalanage,T.；Yen,G.C.；Nguyen,H.X.；Nguyen,M.T.Tyrosinaseinhibitors from the wood of Artocarpus heterophyllus.Journal of NaturalProducts 2012,75,1951-1955.；Daus,M.；Chaithada,P.；Phongpaichit,S.；Watanapokasin,R.；Carroll,A.R.；Mahabusarakam,W.New prenylated dihydrochalconesfrom the leaves of Artocarpus elasticus.Phytochemistry Letters 2017,19,226-230.；Toume,K.；Habu,T.；Arai,M.A.；Koyano,T.；Kowithayakorn,T.；Ishibashi,M.Prenylated flavonoids and resveratrol derivatives isolated from Artocarpuscommunis with the ability to overcome TRAIL resistance.Journal of NaturalProducts 2015,78,103-110.；Yuan,W.J.；Yuan,J.B.；Peng,J.B.；Ding,Y.Q.；Zhu,J.X.；Ren,G.Flavonoids from the roots of Artocarpus heterophyllus.Fitoterapia 2017,117,133-137.]。
菠罗蜜(Artocarpus heterophyllus)为桑科波罗蜜属植物，又名木菠萝或树菠萝，原产于印度，现已广泛引种栽培于世界热带各地，在我国海南、云南、广西以及广东等省区常有栽培。波罗蜜是一种集水果，木本粮食以及珍贵用材于一身的著名的热带经济树种。波罗蜜的果实为热带地区著名水果，由于其是世界上最重的水果，一般重达5～20kg，最重可超过50kg，加之果实肥厚柔软，清甜可口，香味浓郁，故被誉为“热带水果皇后”。波罗蜜以果实和种仁入药，具有生津除烦、解酒醒脾的功效，波罗蜜的果实常用于治疗酒精中毒，波罗蜜种仁常用于治疗产后脾虚气弱，乳少或乳汁不行等。截止目前为止，有关波罗蜜果实中的化学成分及其药理活性的研究报道尚少见。在前期研究中发现波罗蜜果实的90％乙醇提取物的具有显著的抗肿瘤活性和多靶点酪氨酸激酶抑制活性。为了更合理地开发利用波罗蜜这种药食同源植物资源，充分发挥其药用价值、食用价值和经济价值，急需对波罗蜜果实中的化学成分及其生物活性进行了系统研究。
发明内容
本发明的目的是提供一种从波罗蜜果肉果干中分离得到的具有新颖化学结构的异戊烯基取代香豆素类化合物artoheteronin，该化合物具有显著的抗肿瘤活性和与阳性对照药相当的蛋白酪氨酸激酶的抑制活性，可以进一步开发成以蛋白酪氨酸激酶为靶标的抗肿瘤药物。
为了实现上述目的，本发明的技术方案为：提供一种异戊烯基取代香豆素类化合物，化学名为artoheteronin，其化学结构如下：
本发明的另一目的是提供一种异戊烯基取代香豆素类化合物artoheteronin的制备方法，包括以下步骤：
A.将阴干的波罗蜜果肉果干粉碎后用甲醇或75％乙醇溶液提取四次，过滤，收集滤液，再减压浓缩干燥，得醇提取物；
B.将醇提取物加蒸馏水制成混悬液，依次用石油醚和乙酸乙酯进行萃取，石油醚萃取液减压浓缩，得石油醚萃取浸膏；
C.将石油醚萃取浸膏进行柱色谱分离纯化，得到单体化合物artoheteronin。进一步地，上述步骤C具体包括：
(1)石油醚萃取浸膏经硅胶柱色谱分离，分别按体积比83:17、68:32和55:45进行石油醚-乙酸乙酯梯度洗脱，收集体积比为68:32的石油醚-乙酸乙酯洗脱物；
(2)取体积比为68:32的石油醚-乙酸乙酯洗脱物用MCI树脂柱层析去除色素，用体积比为55:45和65:35、85:15的甲醇-水梯度洗脱，收集体积比为65:35的甲醇-水洗脱物；
(3)取体积比为65:35的甲醇-水洗脱物进行ODS柱层析，用体积比为55:45、68:32和82:17的甲醇-水梯度洗脱，收集体积比为68:32的甲醇-水洗脱物进行浓缩；
(4)取浓缩后的甲醇-水洗脱物用制备型高效液相色谱分离，流动相为甲醇-水，体积比68:32，得到单体化合物artoheteronin。
本发明的再一目的在于提供异戊烯基取代香豆素类化合物artoheteronin在制备抗肿瘤药物中的应用，尤其是提供异戊烯基取代香豆素类化合物artoheteronin在制备以蛋白酪氨酸激酶为靶标的靶向抗肿瘤药物方面的应用。
进一步地，所述肿瘤细胞株包括HL-60(人原髓细胞白血病细胞)、A549(人肺癌细胞)、SMMC-7721(人肝癌细胞)、MCF-7(人乳腺癌细胞)和SW480(人结肠癌细胞)等五种肿瘤细胞株。
本发明通过对波罗蜜果肉果干中的化学成分进行系统研究过程中，首次从其石油醚萃取部位中分离鉴定了一个具有新颖化学结构的异戊烯基取代香豆素类化合物artoheteronin。多种体外活性评价结果表明：该化合物具有显著的抗肿瘤活性和与阳性对照药相当的蛋白酪氨酸激酶的抑制活性，可以进一步开发成以蛋白酪氨酸激酶为靶标的抗肿瘤药物。
具体实施方式
下面结合实施例，对本发明的具体实施方式作进一步详细描述。以下实施例用于说明本发明，但不用来限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规实验条件。
实施例一：化合物artoheteronin的制备方法
A.将阴干的波罗蜜果肉果干(10.7kg，海南)粉碎后用75％乙醇溶液冷浸提取四次，每次提取一周，过滤，收集滤液，减压浓缩，得乙醇提取物；
B.将该乙醇提取物加蒸馏水制成混悬液，依次用石油醚和乙酸乙酯萃取，石油醚萃取液经减压浓缩，得石油醚萃取物518.8g；
C.将石油醚萃取物进行柱色谱分离纯化：
(1)将石油醚萃取浸膏用硅胶柱层析进行分离，以石油醚-乙酸乙酯(体积比为83:17、68:32和55:45)为洗脱剂进行梯度洗脱，收集石油醚-乙酸乙酯(体积比68:32)洗脱物；
(2)取石油醚-乙酸乙酯(体积比68:32)洗脱物用MCI树脂柱层析去除色素，用甲醇-水(体积比55:45、65:35和85:15)为洗脱剂梯度洗脱，收集甲醇-水(体积比65:35)洗脱物；
(3)取甲醇-水(体积比65:35)洗脱物进行ODS柱层析，以甲醇-水(体积比55:45、68:32和82:17)为洗脱剂进行梯度洗脱，收集甲醇-水(体积比68:32)洗脱物进行浓缩；
(4)取浓缩后的甲醇-水洗脱物用制备型高效液相色谱分离，流动相为甲醇-水(体积比68:32)，得到纯的化合物artoheteronin(128.6mg)。
结构确证：通过旋光光谱、紫外(UV)光谱、红外(IR)光谱、核磁共振(NMR)谱和质谱(MS)等多种现代波谱技术的综合解析，确定了化合物artoheteronin的化学结构。
Artoheteronin:白色无定形粉末,-18.6(c 0.12,CH3OH)；IR(KBr)vmax3398,3028,2929,1721,1620,1579,1480,1381,1269,1151,1072,958,862,739cm–1；UV(CH3OH)λmax(logε)219(4.42),249(4.02),268(3.81),329(4.18)nm；ESIMS m/z349[M+H]+；HRESIMS m/z 349.1648[M+H]+(Calcd for C19H25O6,349.1646)；1H-NMR(400MHz,CDCl3)δ:7.56(1H,s,H-5),7.49(1H,s,H-4),6.66(1H,s,H-8),6.14(1H,dd,J＝17.2,10.8Hz,H-2'),5.10(1H,d,J＝10.8Hz,H-3'α),5.06(1H,d,J＝17.2Hz,H-3'β),4.61(1H,d,J＝8.9Hz,H-1”),3.65(1H,d,J＝8.9Hz,H-2”),1.53(3H,s,H-4”),1.46(3H,s,H-4'),1.46(3H,s,H-5'),1.26(3H,s,H-5”)；13C NMR(100MHz,CDCl3)δ:160.1(C-2),155.0(C-7),154.2(C-8a),145.5(C-2'),137.7(C-4),131.9(C-3),126.9(C-5),120.9(C-6),113.3(C-4a),112.2(C-3'),103.4(C-8),79.7(C-3”),76.1(C-2”),69.0(C-1”),40.3(C-1'),26.6(C-4”),26.1(C-4'),26.1(C-5'),19.1(C-5”)。
实施例二：化合物artoheteronin的制备方法
A.将阴干的波罗蜜果肉果干(100.8kg，海南)粉碎后用甲醇冷浸提取四次，每次一周，过滤，收集滤液，减压浓缩，得甲醇提取物；
B.将甲醇提取物加蒸馏水制成混悬液，依次用石油醚和乙酸乙酯萃取，石油醚萃取液减压浓缩，得石油醚萃取物5218.7g；
C.将石油醚萃取物进行柱色谱分离纯化：
(1)将石油醚部位浸膏用硅胶柱层析进行分离，石油醚-乙酸乙酯(体积比为83:17、68:32和55:45)为洗脱剂进行梯度洗脱，收集石油醚-乙酸乙酯(体积比68:32)洗脱物；
(2)取石油醚-乙酸乙酯(体积比68:32)洗脱物用MCI树脂柱层析去除色素，以甲醇-水(体积比55:45、65:35和85:15)为洗脱剂进行梯度洗脱，收集甲醇-水(体积比65:35)洗脱物；
(3)取甲醇-水(体积比65:35)洗脱物进行ODS柱层析，以甲醇-水(体积比55:45、68:32和82:17)为洗脱剂进行梯度洗脱，收集甲醇-水(体积比68:32)洗脱物进行浓缩；
(4)取浓缩后的甲醇-水洗脱物用制备型高效液相色谱分离，流动相为甲醇-水(体积比68:32)，得到单体化合物II(1186.6mg)。
化合物II的结构确证：淡黄色无定形粉末；HR-ESI-MS显示化合物II的[M+H]+为m/z349.1648；化合物II与实施例一中制备方法得到的化合物artoheteronin共TLC，在三种展开体系下[石油醚-乙酸乙酯(6:4)、石油醚-丙酮(7:3)和氯仿-丙酮(9:1)]均为均一斑点，说明该化合物与化合物artoheteronin为同一化合物。
一、化合物artoheteronin的抗肿瘤活性研究
1、实验方法：将五种常见肿瘤细胞株HL-60、A549、SMMC-7721、MCF-7和SW480分别用含10％小牛血清的RPMI-1640培养基，在37℃、5％CO2培养箱中培养。采用MTT法进行细胞增殖抑制试验，主要操作为：取对数生长期的肿瘤细胞株，用0.25％的胰蛋白酶消化，10％新生小牛血清的RPMI-1640培养液调制成5×104个/mL的细胞悬液，接种于96孔板中，每孔接种180μL。在37℃,5％CO2饱和湿度条件下培养8-10h，待其贴壁，每个孔加入用PBS配制的样品液，使得样品终浓度分别为0.1、1和10μg/mL。每个浓度平行3孔，继续培养44h后，每孔加入50μL MTT(1mg/mL-1，PBS配制)，在37℃，5％CO2条件下继续温育4h，吸弃孔内培养上清液，每孔加入150μL DMSO，在微型振荡器上摇匀15min，结晶溶解后，在酶联免疫检测仪上选择570nm，测定各孔的吸光值，同时设置空白组(仅加入含细胞的培养液)和对照组(以培养液替代药物)，计算细胞增殖抑制率。抑制率(％)＝(1-实验组3孔OD值平均值/对照组3孔OD值平均值)×100％。以抑制率作纵坐标，作回归曲线，计算出样品IC50值。采用SPSS13.0统计软件包进行数据处理及统计分析。
2、抗肿瘤活性实验结果(见表1)
由本发明实施例一得到的化合物artoheteronin对所选肿瘤细胞株HL-60、A549、SMMC-7721、MCF-7和SW480均显示不同程度的增殖抑制活性。
表1化合物artoheteronin的抗肿瘤活性评价结果
二、化合物artoheteronin的抑制蛋白酪氨酸激酶活性
大鼠脑组织中PTKs的提取：将大鼠大脑取出，剔除脑膜，称重，加入4倍量的冷匀浆液。冰浴中用玻璃匀浆器高速匀浆，离心，收集上清液，再离心10min。收集上清液，上清液中含有胞浆型酪氨酸激酶，而沉淀可作为受体型酪氨酸激酶使用。留取少量上清液用于提取物中蛋白质的含量测定，其余分装，置于-70℃保存备用。
酶标板包被：将底物稀释液加入96孔酶标板中(每孔125μL)，37℃孵育过夜。移除板中过量底物液，加入磷酸盐缓冲液(PBS-Tween20)洗涤，于37℃干燥2h。4℃保存备用。
PTK抑制剂评价：先将样品加入酶标板中，37℃孵育，加入用激酶缓冲液稀释的ATP，37℃孵育，移除板中的反应液，洗涤；加入抗体复合物，37℃孵育；移除板中抗体复合物，洗涤，加入四甲基联苯胺(TMB)显色液，室温避光反应，加入终止液，于450nm波长处测定吸光度(A)值。阳性对照药为伊马替尼。按下述公式计算化合物artoheteronin的抑制率：抑制率％＝(A正常-A样品)/(A正常-A空白)*100％
结果表明，化合物artoheteronin对蛋白酪氨酸激酶具有显著的抑制作用(抑制率81.08％)，抑制活性和阳性对照药伊马替尼的抑制活性相当(抑制率69.96％)。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明技术原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (5)
1.一种异戊烯基取代香豆素类化合物的制备方法，其特征在于：化学名为artoheteronin，其化学结构如下：
；
其制备方法，包括以下步骤：
A. 将阴干的波罗蜜果肉果干粉碎后用甲醇或75%乙醇溶液提取四次，过滤，收集滤液，再减压浓缩干燥，得醇提取物；
B. 将醇提取物加蒸馏水制成混悬液，依次用石油醚和乙酸乙酯进行萃取，石油醚萃取液减压浓缩，得石油醚萃取浸膏；
C. 将石油醚萃取浸膏进行柱色谱分离纯化，得到单体化合物artoheteronin；
步骤C包括：
（1）石油醚萃取浸膏经硅胶柱色谱分离，分别按体积比83:17、68:32和55:45进行石油醚-乙酸乙酯梯度洗脱，收集体积比为68:32的石油醚-乙酸乙酯洗脱物；
（2）取体积比为68:32的石油醚-乙酸乙酯洗脱物用MCI树脂柱层析去除色素，用体积比为55:45和65:35、85:15的甲醇-水梯度洗脱，收集体积比为65:35的甲醇-水洗脱物；
（3）取体积比为65:35的甲醇-水洗脱物进行ODS柱层析，用体积比为55:45、68:32和82:17的甲醇-水梯度洗脱，收集体积比为68:32的甲醇-水洗脱物进行浓缩；
（4）取浓缩后的甲醇-水洗脱物用制备型高效液相色谱分离，流动相为甲醇-水，体积比68:32，得到单体化合物artoheteronin。
2.根据权利要求1所述的异戊烯基取代香豆素类化合物的制备方法制备得到的化合物在制备抗肿瘤药物中的应用。
3.根据权利要求2所述的异戊烯基取代香豆素类化合物的制备方法制备得到的化合物在制备以蛋白酪氨酸激酶为靶标的靶向抗肿瘤药物方面的应用。
4.根据权利要求3所述的异戊烯基取代香豆素类化合物的制备方法制备得到的化合物在制备抗肿瘤药物中的应用，所述肿瘤的肿瘤细胞株包括HL-60、 A549、SMMC-7721、MCF-7或SW480 五种肿瘤细胞株。
5.根据权利要求4所述的异戊烯基取代香豆素类化合物的制备方法制备得到的化合物在制备以蛋白酪氨酸激酶为靶标的靶向抗肿瘤药物方面的应用，所述肿瘤的肿瘤细胞株包括HL-60、 A549、SMMC-7721、MCF-7或SW480 五种肿瘤细胞株。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110745925.9A CN113354609B (zh) | 2021-07-01 | 2021-07-01 | 一种异戊烯基取代香豆素类化合物及其制备方法和应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110745925.9A CN113354609B (zh) | 2021-07-01 | 2021-07-01 | 一种异戊烯基取代香豆素类化合物及其制备方法和应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113354609A CN113354609A (zh) | 2021-09-07 |
| CN113354609B true CN113354609B (zh) | 2024-04-05 |
Family
ID=77538032
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110745925.9A Active CN113354609B (zh) | 2021-07-01 | 2021-07-01 | 一种异戊烯基取代香豆素类化合物及其制备方法和应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113354609B (zh) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108250207A (zh) * | 2018-03-26 | 2018-07-06 | 海南师范大学 | 一种新的香豆素类化合物及其制备方法和应用 |
-
2021
- 2021-07-01 CN CN202110745925.9A patent/CN113354609B/zh active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108250207A (zh) * | 2018-03-26 | 2018-07-06 | 海南师范大学 | 一种新的香豆素类化合物及其制备方法和应用 |
Non-Patent Citations (4)
| Title |
|---|
| In Vitro Cytotoxic Screening of Coumarins;Montagner C等;《Lat. Am. J. Pharm》;第30卷(第09期);第1671-1672页Table 1-2 * |
| LeiTao等.Prenylated coumarins from the fruits of Artocarpus heterophyllus with their potential anti-inflammatory and anti-HIV activities.《Natural Product Research》.2021,第36卷(第10期),第3页Figure 1中的compound 1. * |
| Prenylated coumarins from the fruits of Artocarpus heterophyllus with their potential anti-inflammatory and anti-HIV activities;LeiTao等;《Natural Product Research》;第36卷(第10期);第3页Figure 1中的compound 1 * |
| 波罗蜜果实中化学成分研究;刘艳萍等;《中草药》;第52卷(第23期);第7105-7110页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113354609A (zh) | 2021-09-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | New phenolic compounds from the leaves of Artocarpus heterophyllus | |
| Jia et al. | Non-flavonoid phenolics from Averrhoa carambola fresh fruit | |
| Yang et al. | Bioassay-guided isolation of cyclooxygenase-2 inhibitory and antioxidant phenylpropanoid derivatives from the roots of Dendropanax dentiger | |
| Xu et al. | Flavonoids from the capitula of Eriocaulon australe | |
| Vinh et al. | Identification of potential anti-inflammatory and melanoma cytotoxic compounds from Aegiceras corniculatum | |
| CN108358921B (zh) | 一种新的吲哚生物碱类化合物及其制备方法和应用 | |
| Roza et al. | Flavonoids from cyclopia genistoides and their xanthine oxidase inhibitory activity | |
| Hou et al. | Separation of constituents from Bergenia stracheyi (Hook. F. & Thoms.) Engl. by high‐speed countercurrent chromatography with elution mode and its antidiabetic and antioxidant in vitro evaluation | |
| Wang et al. | Chemical components of the fruits of Morus nigra Linn.: Methyl caffeate as a potential anticancer agent by targeting 3-phosphoglycerate dehydrogenase | |
| Xiang et al. | Screening, characterization of trace α-glucosidase inhibitors from the root of Pueraria lobata and evaluation of their hypoglycemic activity | |
| CN108530282B (zh) | 一种stemodane型二萜类化合物及其制备方法和应用 | |
| CN113354609B (zh) | 一种异戊烯基取代香豆素类化合物及其制备方法和应用 | |
| Chen et al. | Homoisoflavonoids from the tuberous roots of Ophiopogon japonicus with their cytotoxic activities | |
| CN108250207B (zh) | 一种香豆素类化合物及其制备方法和应用 | |
| Wang et al. | Constituents of the fruits of Rubus chingii Hu and their neuroprotective effects on human neuroblastoma SH-SY5Y cells | |
| CN114573456B (zh) | 一种贝壳杉烷型二萜化合物及其制备方法和应用 | |
| Choi et al. | Beneficial role of Boehmeria nivea in health and phytochemical constituents | |
| CN106831775B (zh) | 一种单萜吲哚生物碱类化合物的制备方法及其应用 | |
| CN113321695B (zh) | 一种甾体类化合物及其制备方法和应用 | |
| Cao et al. | NO release inhibitory activity of flavonoids from Aesculus wilsonii seeds through MAPK (P38), NF-κB, and STAT3 cross-talk signaling pathways | |
| CN108822093B (zh) | 一种异戊烯基异黄酮类化合物及其制备方法和应用 | |
| CN108341849A (zh) | 玉蕊醇类三萜化合物及其制备方法和在制药中的用途 | |
| Wang et al. | Optimization of the extraction conditions and simultaneous quantification of six flavonoid glycosides in Flos Chrysanthemi by RP-LC | |
| Liangsakul et al. | Anti-HIV 1 Activity of Xanthones from the Bark of Mammea harmandii | |
| Xia et al. | New Phenolic Glycosides and Lignans from the Roots of Lilium dauricum |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |