Ogasawara Akira, Torimoto Nao, Tsuda Naoki, Aohara Fumika, Ohashi Rikiya, Yamada Yasuhiro, Taniguchi Hideki
Advanced Drug Research Laboratories, Sohyaku Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, 2-2-50 Kawagishi, Toda-shi, Saitama 335-8505, Japan;.
Drug Metab Lett. 2016;10(3):152-160. doi: 10.2174/1872312810666160822110011.
Cytochrome P450 (CYP) enzymes are induced by some therapeutic drugs, leading to interactions reducing drug plasma concentrations. Recently, an assessment of CYP induction using messenger RNA (mRNA) levels has shown advantages over measurement of enzymatic activity; it has a larger dynamic range of induction and enables us to measure the intrinsic induction potential of time-dependent CYP inhibitors. In order to minimize the late-stage attrition of new chemical entities (NCE), it is important to evaluate CYP induction potency at mRNA levels in the early stage of drug development.
The aim of this study is to establish a new screening method to evaluate induction potency of CYP1A2, CYP2B6, and CYP3A4 at mRNA levels.
QuantiGene Plex 2.0 Assay using HepaRG cells.
The results from our new CYP induction assay system corresponded well to the already reported results obtained by using human hepatocytes. The induction potency was evaluated by calculating the concentration of test compounds that gives 10% of positive control response (R10), which is measurable even when full dose-response curves cannot be obtained. Compared with the evaluation of CYP induction in human hepatocytes, the response at R10 in HepaRG cells suggested the possibility of exhibiting induction potency for corresponding CYPs. Interestingly, the results with our in-house 109 compounds showed that several compounds induced CYP1A2 or CYP2B6 expression without upregulation of CYP3A4.
Our developed assay system, as well as the R10 value, is useful for evaluating the CYP induction potency of NCE in early drug discovery.
细胞色素P450（CYP）酶可被某些治疗药物诱导，从而导致相互作用，降低药物血浆浓度。最近，使用信使核糖核酸（mRNA）水平评估CYP诱导作用已显示出优于酶活性测量的优势；它具有更大的诱导动态范围，使我们能够测量时间依赖性CYP抑制剂的内在诱导潜力。为了尽量减少新化学实体（NCE）的后期损耗，在药物开发的早期阶段评估mRNA水平的CYP诱导潜力非常重要。
本研究的目的是建立一种新的筛选方法，以评估CYP1A2、CYP2B6和CYP3A4在mRNA水平的诱导潜力。
使用HepaRG细胞的QuantiGene Plex 2.0检测法。
我们新的CYP诱导检测系统的结果与已报道的使用人肝细胞获得的结果非常吻合。通过计算产生阳性对照反应10%（R10）的受试化合物浓度来评估诱导潜力，即使无法获得完整的剂量-反应曲线，该浓度也是可测量的。与在人肝细胞中评估CYP诱导作用相比，HepaRG细胞中R10处的反应表明受试化合物对相应CYP具有诱导潜力。有趣的是，我们内部的109种化合物的结果表明，几种化合物可诱导CYP1A2或CYP2B6表达，而不会上调CYP3A4。
我们开发的检测系统以及R10值，可用于在早期药物发现中评估NCE的CYP诱导潜力。
