Quick identification of apoptosis inducer from Isodon eriocalyx by a drug discovery platform composed of analytical high-speed counter-current chromatography and the fluorescence-based caspase-3 biosensor detection
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摘要Analytical high-speed counter-current chromatography (HSCCC), a unique liquid-to-liquid separation technology, has an inherent capability to provide perfect fractionation for tracking active ingredients of medicinal herbs, in a quick, efficient, and high-recovery manner. A high throughput screening (HTS) method which utilizes a novel biosensor that selectively detects apoptosis based on the fluorescence resonance energy transfer (FRET) technique, was newly established and proved to be very sensitive in detecting apoptosis induced by various known anticancer drugs. The first combination of both advanced techniques formed an efficient platform for drug discovery and succeeded in quickly identifying the most potent apoptotic constituent of a Chinese herb namely Isodon eriocalyx. The system of n-hexane/ethyl acetate/methanol/water was used as the separation solvent. The solvent ratio was first set at 3:5:3:5 to check the water-soluble part of the crude extract, and then 1:1:1:1 was used to isolate the target compounds. The active fraction was tracked and purified continuously using HSCCC which was guided by the apoptosis detection at gradually decreased drug concentrations. As a result, the most potent apoptosis inducer in this herb was discovered by analytical HSCCC equipped with a 16 ml mini-coil column, using less than 50 ml diphase solvent, from about 50 mg active fraction. It was identified as eriocalyxin B, a well-known antitumor natural product, by NMR analysis of the HSCCC purified fraction. (C) 2010 Elsevier B.V. All rights reserved.
著者Han QB, Yu T, Lai F, Zhou Y, Feng C, Wang WN, Fu XH, Lau CBS, Luo KQ, Xu HX, Sun HD, Fung KP, Leung PC
期刊名稱Talanta
出版年份2010
月份9
日期15
卷號82
期次4
出版社Elsevier
頁次1521 - 1527
國際標準期刊號0039-9140
電子國際標準期刊號1873-3573
語言英式英語
關鍵詞Analytical high-speed counter-current chromatography; Apoptosis inducer; Caspase-3 biosensor; Eriocalyxin B; Isodon eriocalyx
Web of Science 學科類別Chemistry; Chemistry, Analytical; CHEMISTRY, ANALYTICAL

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