| Angiogenesis is critical for the growth, maintenance, and metastasis of tumors. Anti-angiogenesis as a promising anticancer strategy has led to the discovery of many natural and synthetic anticancer agents. Licorice, the roots and stolons of some Glycyrrhiza species, has been used for a long time as one of the most important crude drugs in the world. Numerous studies have confirmed that licorice extract possesses many pharmacological activities such as antioxidant, antibacterial, antiin?ammatory and anticancer activities. However, little is known about the mechanism of antiangiogenesis. The article applied zebrafish angiogenic model for the screening of bioactive constituents from licorice, and the bioactive compounds were isolated by bioassay-guided separation procedure and their structures were identified by spectral analysis.The 70% ethanol extract of licorice was extracted with petroleum ether, ethyl acetate, n-butanol, then the bioactivity of the extracts was detected by zebrafish angiogenic model. The inhibition of the extracts showed that the highest active components existed in ethyl acetate extract.The ethyl acetate extract was fractionationed by polyamide column chromatography to afford 7 fractions(Fr1~Fr7), and Fr5, Fr6 showed antiangiogenic activity.Fr5 and Fr6 were further separated by HSCCC, 12 compounds were obtained and 5 of them were elucidated as following: isoliquiritigenin, isolicoflavonol, licocoumarone, glyurallin A and glyurallin B. Fr1~Fr4 were alse further separated by HSCCC, 8 compounds were obtained and 5 of them were elucidated as following: liquiritin, ononin, isoliquiritin, liquiritigenin, formononetin.The compounds of Fr5 and Fr6 were detected by zebrafish angiogenic model. Results showed that isoliquiritigenin and isolicoflavonol had remarkable inhibitory effect on the angiogenesis of zebrafish. When the concentration of isoliquiritigenin and isolicoflavonol was 10μg/mL, the inhitory rate was 100%.
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