Development of GC-MS based cytochrome P450 assay for the investigation of multi-herb interaction

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• 作者：Hyun-A Oh^a ; ^b ; Hyunbeom Lee^a ; Donghak Kim^b ; Byung Hwa Jung^a ; ^c ; ^{jbhluck@kist.re.kr}
• 关键词：GC-MS ; Human liver microsomes ; Cytochrome P450 ; Cocktail approach ; Socheongryong-tang
• 刊名：Analytical Biochemistry
• 出版年：2017
• 出版时间：15 February 2017
• 年：2017
• 卷：519
• 期：Complete
• 页码：71-83
• 全文大小：3139 K
• 卷排序：519

文摘

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.