Direct and quantitative evaluation of the human CYP3A4 contribution (fm) to drug clearance using the in vitro SILENSOMES model
Direct and quantitative evaluation of the human CYP3A4 contribution (fm) to drug clearance using the in vitro SILENSOMES model
复制标题
DOI：
10.1080/00498254.2016.1208854
复制
发表时间：
2017-01-01
期刊：
影响因子：
1.8
通讯作者：
Walther, Bernard
中科院分区：
文献类型：
--
作者：
Parmentier, Yannick;Pothier, Corinne;Walther, Bernard
1.在监管机构推荐的不同体外研究中，没有一种金标准模型能够方便直接地测量定量的CYP450对药物生物转化的贡献。在这篇文章中，我们提出了一种名为Silensome(TM)的原创策略，由于基于特定机制的抑制剂(MBI)，可以生产针对一个特定CYP450的沉默的人肝微粒体。2.使用Azamrin作为特定的CYP3A4 MBI，我们证明了概念证明，我们的过程可以完全、特异性(甚至相对于3A5)和永久(至少六年)抑制CYP3A4。因此，比较对照和CYP3A4-Silensome(TM)中的清除量，确定了11种与已知的体内贡献相关的CYP3A4底物，并显示出误差小于10%的准确性。相比之下，用重组人细胞色素P450(RhCYP450)测定的贡献率较低(30%的测试底物的误差超过10%)。3.这种简单易用的体外方法结合了现有模型的优点(重组人细胞色素P450的特异性和HLM的代表性)，但没有缺点。同样的策略可以用来逐个沉默其他主要的CYP450，以提供完整的直接CYP450定量表型试剂盒。
1.Among the different in vitro studies recommended by the regulatory agencies, no gold-standard model can easily and directly measure the quantitative CYP450 contributions to drug biotransformation. In this article, we propose an original strategy, called Silensomes(TM), to produce human liver microsomes silenced for one specific CYP450, thanks to specific mechanism-based inhibitors (MBI).2.Using azamulin as a specific CYP3A4 MBI, we demonstrated the proof of concept that CYP3A4 can be totally, specifically (even against 3A5) and permanently (at least for six years) inhibited by our process. Thus, comparing clearance in control and CYP3A4-Silensomes(TM), CYP3A4 contributions were determined for 11 CYP3A4 substrates which correlated with known in vivo contributions and revealed accuracy with less than 10% error. In comparison, contributions determined using recombinant human CYP450 (rhCYP450s) were less accurate (more than 10% error for 30% of the tested CYP3A4 substrates).3.This easy and ready-to-use in vitro method combines the advantages of existing models (specificity of rhCYP450s and representativeness of HLM) without their drawbacks. The same strategy could be used to silence other major CYP450s one-by-one to provide a complete direct CYP450 quantitative phenotyping kit.