The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.
School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, China.
Arch Toxicol. 2023 Aug;97(8):2133-2142. doi: 10.1007/s00204-023-03524-1. Epub 2023 May 20.
In this study, the effects of 17 CYP3A4 variants and drug-drug interactions (DDI) with its mechanism on alectinib metabolism were investigated. In vitro incubation systems of rat liver microsomes (RLM), human liver microsomes (HLM) and recombinant human CYP3A4 variants were established. The formers were used to screen potential drugs that inhibited alectinib metabolism and study the underlying mechanism, and the latter was used to determine the dynamic characteristics of CYP3A4 variants. Alectinib and its main metabolite M4 were quantitatively determined by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The results showed that compared with CYP3A4.1, only CYP3A4.29 showed higher catalytic activity, while the catalytic activity of CYP3A4.4, .7, .8, .12, .14, .16, .17, .18, .19, .20, .23, and .24 decreased significantly. Among them, the catalytic activity of CYP3A4.20 is the lowest, only 2.63% of that of CYP3A4.1. Based on the RLM incubation system in vitro, 81 drugs that may be combined with alectinib were screened, among which 18 drugs had an inhibition rate higher than 80%. In addition, nicardipine had an inhibition rate of 95.09% with a half-maximum inhibitory concentration (IC) value of 3.54 ± 0.96 μM in RLM and 1.52 ± 0.038 μM in HLM, respectively. There was a mixture of non-competitive and anti-competitive inhibition of alectinib metabolism in both RLM and HLM. In vivo experiments of Sprague-Dawley (SD) rats, compared with the control group (30 mg/kg alectinib alone), the AUC, AUC, T and C of alectinib administered in combination with 6 mg/kg nicardipine were significantly increased in the experimental group. In conclusion, the metabolism of alectinib was affected by polymorphisms of the CYP3A4 gene and nicardipine. This study provides reference data for clinical individualized administration of alectinib in the future.
本研究考察了 17 种 CYP3A4 变体及其与药物相互作用（DDI）对艾乐替尼代谢的影响及其机制。建立了大鼠肝微粒体（RLM）、人肝微粒体（HLM）和重组人 CYP3A4 变体的体外孵育系统。前者用于筛选可能抑制艾乐替尼代谢的潜在药物，并研究其潜在机制，后者用于确定 CYP3A4 变体的动力学特征。采用超高效液相色谱串联质谱法（UPLC-MS/MS）定量测定艾乐替尼及其主要代谢产物 M4。结果表明，与 CYP3A4.1 相比，只有 CYP3A4.29 表现出更高的催化活性，而 CYP3A4.4、.7、.8、.12、.14、.16、.17、.18、.19、.20、.23 和.24 的催化活性显著降低。其中，CYP3A4.20 的催化活性最低，仅为 CYP3A4.1 的 2.63%。基于体外 RLM 孵育系统，筛选出可能与艾乐替尼合用的 81 种药物，其中 18 种药物的抑制率高于 80%。此外，尼卡地平在 RLM 和 HLM 中的抑制率分别为 95.09%和 1.52±0.038μM，IC 值分别为 3.54±0.96μM 和 1.52±0.038μM。在 RLM 和 HLM 中，艾乐替尼代谢均存在混合的非竞争性和反竞争性抑制。Sprague-Dawley（SD）大鼠体内实验结果表明，与对照组（单独 30mg/kg 艾乐替尼）相比，实验组联合 6mg/kg 尼卡地平给药后，艾乐替尼的 AUC、AUC、T 和 C 均显著增加。综上所述，CYP3A4 基因多态性和尼卡地平均可影响艾乐替尼的代谢。本研究为今后临床个体化应用艾乐替尼提供了参考数据。
