title: Rapid screening and identification of α-amylase inhibitors from Garcinia xanthochymus using enzyme-immobilized magnetic nanoparticles coupled with HPLC and MS.
abstract: α-Amylase inhibitors play an important role in management of diabetes and obesity. In order to rapidly discover potent α-amylase inhibitors from medicinal plants, a ligands-screening method based on enzyme-immobilized magnetic nanoparticles integrated with HPLC was developed. Amine-terminated magnetic nanoparticles were prepared for the immobilization of α-amylase. Based on the affinity theory, the α-amylase-coated magnetic nanoparticles were employed to fish out the ligands from the extracts of Garcinia xanthochymus, and the elutes were examined by HPLC. As a result, three ligands were screened out. Isolation and identification were carried out subsequently. By analyzing the UV, MS and NMR spectra, they were identified as three biflavonoids including GB2a glucoside (2), GB2a (3) and fukugetin (4). The IC50 values of the three compounds were also determined. The results suggest the proposed approach is efficient and accurate, and has great potential in rapid discovery of drug candidates from medical plants.
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China; College of Pharmacy, South Central University for Nationalities, Wuhan 430074, China.
authors.forename: Yunfang
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Chemistry and Material Sciences, South Central University for Nationalities, Wuhan 430074, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Chen
authors.affiliation: College of Pharmacy, South Central University for Nationalities, Wuhan 430074, China.
authors.forename: Chuying
authors.initials: C
authors.lastname: Xiao
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Chen
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Ministry of Education), and School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China. Electronic address: yuxiuxiao2011@whu.edu.cn.
authors.forename: Yuxiu
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: College of Pharmacy, South Central University for Nationalities, Wuhan 430074, China. Electronic address: meizhinan@163.com.
authors.forename: Zhinan
authors.initials: Z
authors.lastname: Mei
mesh_terms: D044946:Biflavonoids / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D029760:Garcinia / Q000737:chemistry*; D058185:Magnetite Nanoparticles / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D010936:Plant Extracts / Q000737:chemistry; D000516:alpha-Amylases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*
chemical_list: D044946:Biflavonoids; D004800:Enzymes, Immobilized; D058185:Magnetite Nanoparticles; D010936:Plant Extracts; D000516:alpha-Amylases
keywords: Garcinia xanthochymus; Ligand fishing; Magnetic nanoparticles; Screening; α-Amylase inhibitors