Tracking antiangiogenic components from G lycyrrhiza uralensis F isch. based on zebrafish assays using high‐speed countercurrent chromatography

• Published in: Journal of separation science 2012-05, Vol.35 (9), p.1167-1172
• Main Authors: Han, Liwen, Yuan, Yanqiang, Zhao, Liang, He, Qiuxia, Li, Yubo, Chen, Xiqiang, Liu, Xiuhe, Liu, Kechun
• Format: Article
• Language: English
• ISSN: 1615-9306; 1615-9314
• DOI: 10.1002/jssc.201101031

Natural products are some of the most important sources of lead compounds for drug discovery. The advanced isolation technique of lead compounds of natural origin using therapeutically relevant bioassays is capable of enhancing work efficiency from complex multiconstituent extracts. In the present study, a bioassay‐guided isolation strategy combined with bioactivity screening was used to identify novel angiogenesis inhibitors from licorice ( G lycyrrhiza uralensis F isch.) based on the zebrafish model and rapid preparative separation by high‐speed countercurrent chromatography. Zebrafish embryos at 24 h postfertilization were chosen as the angiogenesis inhibition model for bioactivity screening. A solvent system ( n ‐hexane–ethyl acetate–methanol–water) with different ratios was optimized and applied in the high‐speed countercurrent chromatography separation of two fractions, F r5 and F r6, from the ethyl acetate extract of licorice. Blood circulation and vascular outgrowth in intersegmental vessels were found to be simultaneously inhibited by isoliquiritigenin and isolicoflavonol in a dose‐dependent manner. Thus, these two compounds were identified and considered as active inhibitors against angiogenesis. These experimental results indicate that zebrafish bioassays combined with high‐speed countercurrent chromatography may provide an alternative pathway for the rapid isolation of bioactive natural products.