Using immobilized G-protein coupled receptors to screen bioactive traditional Chinese medicine compounds with multiple targets.
Using immobilized G-protein coupled receptors to screen bioactive traditional Chinese medicine compounds with multiple targets.
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DOI：
10.1016/j.jpba.2012.05.004
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发表时间：
2012-11
影响因子：
3.4
通讯作者：
Xinfeng Zhao;Qian Li;Liujiao Bian;Xiaohui Zheng;Jianbin Zheng;You-yi Zhang;Zi-jian Li
中科院分区：
文献类型：
--
作者：
Xinfeng Zhao;Qian Li;Liujiao Bian;Xiaohui Zheng;Jianbin Zheng;You-yi Zhang;Zi-jian Li
由于市场上新药的减少，如今对用于多靶点化合物筛选的高通量方法的需求继续增加。将两种G蛋白偶联受体--α1-肾上腺素能受体(α1A-AR)和β2-肾上腺素能受体(β2-AR)纯化后固定在大孔硅胶表面，制备了新型色谱固定相。使用对照药物(例如，哌唑嗪、特拉唑嗪、沙丁胺醇和特布他林)来表征所获得的α1A-AR和β2-AR柱的保留行为。本研究还将两个色谱柱与一个六通开关阀耦合，构建了一个用于复杂混合物中多目标化合物筛选的自动二维系统。以盐酸肾上腺素为代表药物对该二维体系的层析性能进行了评价。同时对丹参、黄连的水提物进行了全自动分析。丹参中的化合物不与α1A-AR和β2-AR柱结合。但黄连素、巴马汀和药根碱被筛选为中药中的生物活性化合物，针对这两种受体。所提出的方法是从复杂的基质中识别和分离针对不同蛋白质的化合物的一种替代方法。
Demand on high-throughput methods for multi-target compounds screening continues to increase nowadays due to the decline of new drugs on the market. Two kinds of G-protein-coupled receptors, alpha1-adrenoceptor (α1A-AR) and beta2-adrenoceptor (β2-AR), were purified and immobilized on the surface of macroporous silica gel to prepare new chromatographic stationary phases. Control drugs (e.g., prazosin, terazosin, salbutamol, and terbutaline) were used to characterize the retention behavior of the obtained α1A-AR and β2-AR columns. This study also coupled both columns with a six-way switching valve to construct an automatic two-dimensional system for multi-target compounds screening in complex mixtures. Adrenaline hydrochloride was used as a representative drug to evaluate the chromatographic performance of the two dimensional system. The aqueous extracts from Salvia miltiorrhiza and Coptis chinensis were also analyzed by the automatic system. The compounds in S. miltiorrhiza had no binding to both α1A-AR and β2-AR columns. But berberine, palmatine and jatrorrhizine were screened as the bioactive compounds in C. chinensis, targeting both the receptors. The proposed method is an alternative for recognizing and separating the compounds targeting different proteins from a complex matrix.