title: Development of GC-MS based cytochrome P450 assay for the investigation of multi-herb interaction.
abstract: As drug interactions with cytochrome P450 enzymes become increasingly important in the field of drug discovery, a high-throughput screening method for analysing the effects of a drug is needed. We have developed a simple and rapid simultaneous analytical method using a cocktail approach for measuring the activities of seven cytochrome P450 enzymes (CYP1A2, CYP2A6, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A4). Human liver microsomes were used as a source for the seven cytochrome P450 enzymes, and a gas chromatography-mass spectrometry (GC-MS) was used for analysing their activities. Kinetic studies and inhibition assays of CYP enzymes were performed using known substrates and inhibitors for validating and comparing the reaction rates and time-dependent activities between methods using each substrate versus a method using a cocktail solution. The optimized cocktail method was successfully applied to evaluate the effects of the decoction of Socheongryong-tang (SCRT) on cytochrome P450 enzymes. Our cocktail method provides a simultaneous high-throughput activity assay using GC-MS for the first time. This method is applicable for analysing the drug interactions of various plant-derived mixtures.
authors.affiliation: Molecular Recognition Research Center, Korea Institute of Science and Technology, Seoul 136-791, Republic of Korea; Department of Biological Sciences, Konkuk University, Seoul 143-701, Republic of Korea.
authors.forename: Hyun-A
authors.initials: HA
authors.lastname: Oh
authors.affiliation: Molecular Recognition Research Center, Korea Institute of Science and Technology, Seoul 136-791, Republic of Korea.
authors.forename: Hyunbeom
authors.initials: H
authors.lastname: Lee
authors.affiliation: Department of Biological Sciences, Konkuk University, Seoul 143-701, Republic of Korea.
authors.forename: Donghak
authors.initials: D
authors.lastname: Kim
authors.affiliation: Molecular Recognition Research Center, Korea Institute of Science and Technology, Seoul 136-791, Republic of Korea; University of Science and Technology (UST), Daejeon 305-600, Republic of Korea. Electronic address: jbhluck@kist.re.kr.
authors.forename: Byung Hwa
authors.initials: BH
authors.lastname: Jung
mesh_terms: D001681:Biological Assay / Q000379:methods*; D065607:Cytochrome P-450 Enzyme Inhibitors / Q000494:pharmacology*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000378:metabolism*; D055808:Drug Discovery; D004347:Drug Interactions; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007700:Kinetics; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System
keywords: Cocktail approach; Cytochrome P450; GC-MS; Human liver microsomes; Socheongryong-tang