Using immobilized G-protein coupled receptors to screen bioactive traditional Chinese medicine compounds with multiple targets.
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 (α(1A)-AR) and beta2-adrenoceptor (β(2)-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 α(1A)-AR and β(2)-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 α(1A)-AR and β(2)-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.
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, Northwest University, Xi'an 710069, China.
Xinfeng
X
Zhao
Qian
Q
Li
Liujiao
L
Bian
Xiaohui
X
Zheng
Jianbin
J
Zheng
Youyi
Y
Zhang
Zijian
Z
Li
D018663:Adrenergic Agents / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D001331:Automation; D002138:Calibration; D002846:Chromatography, Affinity / Q000592:standards; D039323:Coptis / Q000737:chemistry; D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods* / Q000592:standards; D006801:Humans; D010946:Plants, Medicinal; D016062:Porosity; D011485:Protein Binding; D011817:Rabbits; D018340:Receptors, Adrenergic, alpha-1 / Q000737:chemistry* / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry* / Q000378:metabolism; D012015:Reference Standards; D027561:Salvia miltiorrhiza / Q000737:chemistry; D058428:Silica Gel / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization
C547713:ADRA1A protein, human; D018663:Adrenergic Agents; D004365:Drugs, Chinese Herbal; D018340:Receptors, Adrenergic, alpha-1; D018343:Receptors, Adrenergic, beta-2; D058428:Silica Gel