Specificity of substrate and inhibitor probes for cytochrome P450s: Evaluation of in vitro metabolism using cDNA-expressed human P450s and human liver microsomes
Specificity of substrate and inhibitor probes for cytochrome P450s: Evaluation of in vitro metabolism using cDNA-expressed human P450s and human liver microsomes
复制标题
DOI：
10.3109/00498259609046742
复制
发表时间：
1996-07-01
期刊：
影响因子：
1.8
通讯作者：
Tsutsui, M
中科院分区：
文献类型：
--
作者：
Ono, S;Hatanaka, T;Tsutsui, M
1.我们用重组牛痘病毒在HepG 2细胞和人肝微粒体中表达的9种人P450形式评价了细胞色素P450的15种底物和14种抑制剂的特异性.香豆素、7-乙氧基试卤灵、7-苄氧基试卤灵、甲苯磺丁脲、苯胺和地西泮分别是CYP 2A 6、CYP 1A亚家族、CYP 2B 6、CYP 2C亚家族、CYP 2 E1和CYP 3A亚家族的形式选择性底物。然而，在所测试的化学品中未发现CYP 2D 6的选择性底物。SKF-525 A抑制所有测试底物的> 40%的代谢活性，并且抑制作用在P450形式之间存在差异。磺胺苯吡唑、7，8-苯甲酮、奎尼丁和醋竹桃霉素分别是CYP 2C亚家族（CYP 2C 19除外）、CYP 1A亚家族、CYP 2D 6和CYP 3A亚家族的选择性抑制剂。Methoxsalen（CYP 2A 6抑制剂）抑制CYP 1A 2和CYP 2A 6的代谢活性。二乙基二硫代氨基甲酸酯（CYP 2 E1抑制剂）除抑制CYP2E1.4的代谢活性外，还抑制CYP 2A 6和CYP 2C 19的代谢活性。我们的研究结果表明，底物和抑制剂报告为P450选择性探针不一定是特定的个人P450的形式。这些结果可能提供有用的信息，在体外使用人肝微粒体样品的人P450底物和抑制剂。
1. We evaluated the specificity of 15 substrates and 14 inhibitors of the cytochrome P450s using nine human P450 forms expressed in HepG2 cells using a recombinant vaccinia virus and also in human liver microsomes.2. Coumarin, 7-ethoxyresorufin, 7-benzyloxyresorufin, tolbutamide, aniline and diazepam were form-selective substrates towards CYP2A6, the CYP1A subfamily, CYP2B6, the CYP2C subfamily, CYP2E1 and the CYP3A subfamily respectively. However, a selective substrate for CYP2D6 was not found among the chemicals tested.3. SKF-525A inhibited > 40 % of the metabolic activity of all substrates tested, and the inhibitory effects differed among P450 forms. Sulphaphenazole, 7,8-benzoflavone, quinidine and troleandomycin were selective inhibitors of the CYP2C subfamily (except CYP2C19), the CYP1A subfamily, CYP2D6 and the CYP3A subfamily respectively. Methoxsalen (CYP2A6 inhibitor) inhibited the metabolic activity of CYP1A2 as well as that of CYP2A6. Diethyldithiocarbamate (CYP2E1 inhibitor) inhibited the metabolic activities of CYP2A6 and CYP2C19 in addition to that of CYP2E1.4. Our results indicated that substrates and inhibitors reported as P450 selective probes are not necessarily specific for individual human P450 forms. These results may provide useful information regarding human P450 substrates and inhibitors in vitro using human liver microsomal samples.