Electrochemically Driven Drug Metabolism by Membranes Containing Human Cytochrome P450
Electrochemically Driven Drug Metabolism by Membranes Containing Human Cytochrome P450
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DOI：
10.1021/ja809364r
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发表时间：
2009-05-20
影响因子：
15
通讯作者：
Komatsu, Yasuo
中科院分区：
文献类型：
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作者：
Mie, Yasuhiro;Suzuki, Masaaki;Komatsu, Yasuo
人细胞色素 P450 (CYP) 代谢 95% 的现有药物，对药物代谢反应的快速分析对于药物开发过程和有效治疗非常重要。由于 CYP 需要电子来代谢药物，因此从电极提供的电子来激活 CYP 分子预计对于开发 CYP 反应的快速测定方法非常有用。尽管已经报道了一些关于分离和纯化的人CYP的直接电化学的研究，但使用含有CYP的微粒体（膜）更合适，因为它们由于易于制备和低成本而经常用于药物研究。在此，我们展示了从电极向涂有疏水薄膜的电极上含有 CYP 和 CYP 还原酶 (CPR) 的微粒体提供的电子，并首次通过伏安法观察电化学驱动的药物代谢。我们测试了微粒体在涂有几种硫醇盐的金电极上的固定化，发现固定在芳香族化合物的薄疏水表面上的微粒体显示出明确定义的氧化还原峰。此外，通过伏安法清楚地观察到CYP3A4的电化学驱动的药物代谢反应，并且这些反应被CYP3A4抑制剂酮康唑抑制。我们还发现心肺复苏可以促进新陈代谢反应。
Rapid analyses of drug metabolism reactions by human cytochrome P450s (CYPs) that metabolize 95% of all current drugs are very important in drug development processes and effective therapies. Since CYPs need electrons to metabolize drugs, electrons supplied from electrodes to activate CYP molecules are expected to be very useful to develop rapid assay methods for CYP reactions. Although several studies on the direct electrochemistry of isolated and purified human CYPs have been reported, the use of microsomes (membranes) containing CYP is more suitable, because they are frequently used in drug research due to their easy preparation and low cost. Herein, we demonstrate electrons supplied from an electrode to microsomes containing CYP and CYP-reductase (CPR) on an electrode coated with hydrophobic thin films and observe electrochemically driven drug metabolisms by voltammetry for the first time. We tested the immobilization of microsomes on gold electrodes coated with several thiolates and found that microsomes immobilized on thin hydrophobic surfaces of aromatic compounds showed welt-defined redox peaks. Furthermore, electrochemically driven drug metabolism reactions of CYP3A4 were clearly observed by voltammetry, and these reactions were inhibited by a CYP3A4 inhibitor, ketoconazole. We also found that metabolism reactions were facilitated in the presence of CPR.