Filppula Anne M, Neuvonen Pertti J, Backman Janne T
Department of Clinical Pharmacology, University of Helsinki, Helsinki, Finland (A.M.F., P.J.N., J.T.B.) and HUSLAB, Helsinki University Central Hospital, Helsinki, Finland (P.J.N., J.T.B.).
Department of Clinical Pharmacology, University of Helsinki, Helsinki, Finland (A.M.F., P.J.N., J.T.B.) and HUSLAB, Helsinki University Central Hospital, Helsinki, Finland (P.J.N., J.T.B.)
Drug Metab Dispos. 2014 Jul;42(7):1202-9. doi: 10.1124/dmd.114.057695. Epub 2014 Apr 8.
Previous studies have shown that several protein kinase inhibitors are time-dependent inhibitors of cytochrome P450 (CYP) 3A. We screened 14 kinase inhibitors for time-dependent inhibition of CYP2C8 and CYP3A. Amodiaquine N-deethylation and midazolam 1'-hydroxylation were used as marker reactions for CYP2C8 and CYP3A activity, respectively. A screening, IC50 shift, and mechanism-based inhibition were assessed with human liver microsomes. In the screening, bosutinib isomer 1, crizotinib, dasatinib, erlotinib, gefitinib, lestaurtinib, nilotinib, pazopanib, saracatinib, sorafenib, and sunitinib exhibited an increased inhibition of CYP3A after a 30-min preincubation with NADPH, as compared with no preincubation. Axitinib and vandetanib tested negative for time-dependent inhibition of CYP3A and CYP2C8, and bosutinib was the only inhibitor causing time-dependent inhibition of CYP2C8. The inhibitory mechanism by bosutinib was consistent with weak mechanism-based inhibition, and its inactivation variables, inhibitor concentration that supports half-maximal rate of inactivation (KI) and maximal inactivation rate (kinact), were 54.8 µM and 0.018 1/min. As several of the tested inhibitors were reported to cause mechanism-based inactivation of CYP3A4 during the progress of this work, detailed experiments with these were not completed. However, lestaurtinib and saracatinib were identified as mechanism-based inhibitors of CYP3A. The KI and kinact of lestaurtinib and saracatinib were 30.7 µM and 0.040 1/min, and 12.6 µM and 0.096 1/min, respectively. Inhibition of CYP2C8 by bosutinib was predicted to have no clinical relevance, whereas therapeutic lestaurtinib and saracatinib concentrations were predicted to increase the plasma exposure to CYP3A-dependent substrates by ≥2.7-fold. The liability of kinase inhibitors to affect CYP enzymes by time-dependent inhibition may have long-lasting consequences and result in clinically relevant drug-drug interactions.
以往研究表明，几种蛋白激酶抑制剂是细胞色素P450（CYP）3A的时间依赖性抑制剂。我们筛选了14种激酶抑制剂对CYP2C8和CYP3A的时间依赖性抑制作用。分别采用阿莫地喹N-去乙基化和咪达唑仑1'-羟化作为CYP2C8和CYP3A活性的标记反应。用人肝微粒体评估筛选、IC50变化及基于机制的抑制作用。在筛选过程中，与未预孵育相比，波舒替尼异构体1、克唑替尼、达沙替尼、厄洛替尼、吉非替尼、来他替尼、尼洛替尼、帕唑帕尼、萨拉替尼、索拉非尼和舒尼替尼在与NADPH预孵育30分钟后对CYP3A的抑制作用增强。阿昔替尼和凡德他尼对CYP3A和CYP2C8的时间依赖性抑制检测为阴性，波舒替尼是唯一引起CYP2C8时间依赖性抑制的抑制剂。波舒替尼的抑制机制与弱的基于机制的抑制作用一致，其失活变量，即支持半数最大失活速率的抑制剂浓度（KI）和最大失活速率（kinact）分别为54.8 μM和0.018 1/分钟。由于在本研究过程中有几种受试抑制剂据报道会引起CYP3A4的基于机制的失活，因此未完成对这些抑制剂的详细实验。然而，来他替尼和萨拉替尼被确定为CYP3A的基于机制的抑制剂。来他替尼和萨拉替尼的KI和kinact分别为30.7 μM和0.040 1/分钟，以及12.6 μM和0.096 1/分钟。预计波舒替尼对CYP2C8的抑制作用无临床相关性，而来他替尼和萨拉替尼的治疗浓度预计会使CYP3A依赖性底物的血浆暴露增加≥2.7倍。激酶抑制剂通过时间依赖性抑制影响CYP酶的可能性可能会产生长期后果，并导致具有临床相关性的药物相互作用。
