Tracking antiangiogenic components from Glycyrrhiza uralensis Fisch. based on zebrafish assays using high-speed countercurrent chromatography

• Published in: Journal of separation science Vol. 35; no. 9; pp. 1167 - 1172
• Main Authors: Han, Liwen, Yuan, Yanqiang, Zhao, Liang, He, Qiuxia, Li, Yubo, Chen, Xiqiang, Liu, Xiuhe, Liu, Kechun
• Format: Journal Article
• Language: English
• Published: Weinheim Blackwell Publishing Ltd 01.05.2012; Wiley; Wiley Subscription Services, Inc
• Subjects: Angiogenesis; Angiogenesis Inhibitors - isolation & purification; Angiogenesis Inhibitors - pharmacology; Animals; Bioassay; Bioassays; Biochemistry; Biological activity; Biological and medical sciences; Biological Assay - methods; Chromatography; Countercurrent Distribution - methods; Danio rerio; Freshwater; General pharmacology; Glycyrrhiza; Glycyrrhiza glabra; Glycyrrhiza uralensis - chemistry; Glycyrrhiza uralensis Fisch; High speed; High-speed countercurrent chromatography (HSCCC); Inhibitors; Isolicoflavonol; Isoliquiritigenin; Lead compounds; Licorice; Medical sciences; Microbiology; Pharmacognosy. Homeopathy. Health food; Pharmacology. Drug treatments; Plant Extracts - isolation & purification; Plant Extracts - pharmacology; Separation; Zebrafish; Zebrafish - growth & development; Asia; China; Antineoplastic agent; Countercurrent chromatography; Food additive; Flavonoid; Medicinal plant; Folk medicine; Dicotyledones; Pisces; Angiospermae; Antiangiogenic agent; Glycyrrhiza uralensis Fisch; Below ground plant part; Isoliquiritigenin; Purification; Isolicoflavonol; Root; Pharmacognosy; Preparative chromatography; Zebrafish; Licorice; High-speed countercurrent chromatography (HSCCC); Biological activity; Vertebrata; Brachydanio rerio; Leguminosae; Glycyrrhiza glabra; Plant origin; Isolation; Flavoring crop; Spermatophyta
• 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 (Glycyrrhiza uralensis Fisch.) 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, Fr5 and Fr6, 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.