A Comprehensive Study on the Effect of Human Genetic Polymorphism on the Drug Metabolizing, Properties of CYP

人类基因多态性对药物代谢及CYP性质影响的综合研究

• 批准号: 18390013
• 负责人: UNO Tadayuki
• 金额: $ 11.14万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18390013/
• 关键词: Cvtochrome P450; Drug Metabolism; Single Nucleotide Polymorphism; Resonance Raman Spectroscopy

In this study, we focused on cytochrome P450 (CYP) which is a dominant group in the human drug metabolizing enzymes, and we aimed at the establishment of a clinical principle which is applicable to order-made treatment, through comprehensive studies on the drug binding and metabolizing activities of mutants which are prepared on the bases of the information on human genomic single-nucleotide polymorphisms (SNPs). We prepared CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4, along with mutants of these CYPs based on the SNP information, and measured drug metabolizing properties of these CYPs. In addition, we measured resonance Raman spectra of them in order to clarify structural factors that are affected by SNPs. Furthermore, we measured drug metabolizing properties of the mutants in order to investigate the effect of SNPs. We could establish that a parameter Kd, which is an index of drug binding affinity, is linearly correlated with Km, a measure of metabolizing activity. In addition, Vmax was found to linearly correlate with the content of 5-coordinated heme. Because the value Vmax/Km is correlated with drug clearance, it is now clear that static values of Kd and 5-coordination reveals the drug clearance. On the other hand, we succeeded in the preparation of CYP3A4 and CYP2D6 by adding a substrate drug to stabilize the protein, and modifying surfactants during the purification step. Thus, we finally established that drug metabolism by CYPs can be conprehensively investigated using these CYPs.

在这项研究中，我们专注于细胞色素P450（CYP），这是人类药物代谢酶的主要群体，我们的目的是建立一种适用于订购治疗的临床原理，该原则是通过对pro依基于人类基因组的突变体进行的，该临床治疗的全面研究。我们根据SNP信息制备了CYP1A2，CYP2C9，CYP2C19，CYP2C19，CYP2D6和CYP3A4，以及这些CYPS的突变体，并测量了这些CYP的药物代谢特性。此外，我们测量了它们的共振拉曼光谱，以阐明受SNP影响的结构因素。此外，我们测量了突变体的药物代谢特性，以研究SNP的作用。我们可以确定，参数KD是药物结合亲和力的指数，与KM线性相关，KM是代谢活性的度量。此外，发现Vmax与5配位血红素的含量线性相关。由于Vmax/Km的值与药物清除相关，因此现在很明显，KD和5配位的静态值揭示了药物清除率。另一方面，我们通过添加底物药物来稳定蛋白质并在纯化步骤中修饰表面活性剂来制备CYP3A4和CYP2D6。因此，我们最终确定CYP可以使用这些CYP对CYP进行药物代谢。

项目成果

Correlations between SNPs and Drug Petabolizing Properties of Human CYPs

SNP 与人类 CYP 药物代谢特性之间的相关性

• 发表时间: 2007
• 作者: Tadayuki;Uno;Yusuke;Kagawa;Ryosuke;Watanabe;Teruaki;Shigetomi;Noritsugu;Ueda;Yoshikazu;Tomisugi;Yoshinobu;Ishikawa;Hatsuo;Maeda
• 通讯作者: Maeda

ヒト薬物代謝酵素CYP2C9の一塩基多型と薬物代謝活性との相関

人药物代谢酶CYP2C9单核苷酸多态性与药物代谢活性的相关性

• 发表时间: 2007
• 作者: 宇野公之;香川雄輔;渡辺亮介;繁富輝明;植田哲嗣;富杉佳計;石川吉伸;前田初男
• 通讯作者: 前田初男