Department of Pharmacy, Faculty of Science (W.K.T., A.R.Y.T., P.S., E.J.H.G., Z.P.Y., N.F.S., A.J.L.) and Department of Pharmacology, Yong Loo Lin School of Medicine (A.J.L.), National University of Singapore, Singapore, Singapore; and Institute of Life Science, Swansea University Medical School, Swansea, United Kingdom (K.A.-M., J.G.L.M.).
Department of Pharmacy, Faculty of Science (W.K.T., A.R.Y.T., P.S., E.J.H.G., Z.P.Y., N.F.S., A.J.L.) and Department of Pharmacology, Yong Loo Lin School of Medicine (A.J.L.), National University of Singapore, Singapore, Singapore; and Institute of Life Science, Swansea University Medical School, Swansea, United Kingdom (K.A.-M., J.G.L.M.)
J Pharmacol Exp Ther. 2020 Aug;374(2):295-307. doi: 10.1124/jpet.120.265249. Epub 2020 May 11.
Gefitinib and erlotinib are epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) with activity against metastatic non-small cell lung cancer. Aldehyde oxidase-1 (AOX1) is a cytosolic drug-metabolizing enzyme. We conducted an experimental and molecular docking study on the effect of gefitinib, erlotinib, and select metabolites on the in vitro catalytic activity of AOX1, as assessed by carbazeran 4-oxidation, and determined the impact of AOX1 inhibition on hepatic metabolism of zaleplon and methotrexate. Gefitinib, desmorpholinopropylgefitinib, erlotinib, desmethylerlotinib, and didesmethylerlotinib inhibited human hepatic cytosolic carbazeran 4-oxidation by a competitive mode, with inhibition constants in submicromolar or low micromolar concentrations. Desmethylgefitinib did not affect AOX1 catalytic activity. A similar pattern was obtained when investigated with human kidney cytosol or recombinant AOX1. The differential effect of gefitinib on human, rat, and mouse hepatic AOX1 catalytic activity suggests species-dependent chemical inhibition of AOX1. Erlotinib was considerably more potent than gefitinib in decreasing hepatic cytosolic zaleplon 5-oxidation and methotrexate 7-oxidation. Molecular docking analyses provided structural insights into the interaction between EGFR-TKIs and AOX1, with key residues and bonds identified, which provided favorable comparison and ranking of potential inhibitors. Based on the US Food and Drug Administration guidance to assess the risk of drug-drug interactions, the calculated R values indicate that further investigations are warranted to determine whether gefitinib and erlotinib impact AOX1-mediated drug metabolism in vivo. Overall, erlotinib desmethylerlotinib, didesmethylerlotinib, gefitinib, and desmorpholinopropylgefitinib are potent inhibitors of human AOX1 catalytic function and hepatic metabolism of zaleplon and methotrexate, potentially affecting drug efficacy or toxicity. SIGNIFICANCE STATEMENT: As epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs), gefitinib and erlotinib are first-line pharmacotherapy for metastatic non-small cell lung cancer. Our experimental findings indicate that clinically relevant concentrations of gefitinib, desmorpholinopropylgefitinib, erlotinib, desmethylerlotinib, and didesmethylerlotinib, but not desmethylgefitinib, inhibit human aldehyde oxidase (AOX1) catalytic activity and hepatic cytosolic metabolism of zaleplon and methotrexate. Molecular docking analysis provide structural insights into the key AOX1 interactions with these EGFR-TKIs. Our findings may trigger improved strategies for new EGFR-TKI design and development.
吉非替尼和厄洛替尼是表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKIs），对转移性非小细胞肺癌具有活性。醛氧化酶 1（AOX1）是一种细胞溶质药物代谢酶。我们进行了一项实验和分子对接研究，以评估吉非替尼、厄洛替尼和选定代谢物对 AOX1 的体外催化活性的影响，通过 carbazeran 4-氧化来评估，以及确定 AOX1 抑制对扎来普隆和甲氨蝶呤肝代谢的影响。吉非替尼、去甲氧基丙基吉非替尼、厄洛替尼、去甲基厄洛替尼和二去甲基厄洛替尼以竞争性方式抑制人肝胞质 carbazeran 4-氧化，抑制常数在亚毫摩尔或低毫摩尔浓度范围内。去甲氧基吉非替尼不影响 AOX1 催化活性。用人肾胞质或重组 AOX1 进行类似的研究也得到了类似的结果。吉非替尼对人、大鼠和小鼠肝 AOX1 催化活性的不同影响表明，AOX1 存在种属依赖性的化学抑制。厄洛替尼比吉非替尼更能降低肝胞质中扎来普隆 5-氧化和甲氨蝶呤 7-氧化。分子对接分析提供了 EGFR-TKIs 与 AOX1 相互作用的结构见解，确定了关键残基和键，这为潜在抑制剂提供了有利的比较和排序。根据美国食品和药物管理局评估药物相互作用风险的指导，计算的 R 值表明，需要进一步研究以确定吉非替尼和厄洛替尼是否会影响体内 AOX1 介导的药物代谢。总的来说，厄洛替尼、去甲基厄洛替尼、二去甲基厄洛替尼、吉非替尼和去甲氧基丙基吉非替尼是人类 AOX1 催化功能和扎来普隆和甲氨蝶呤肝代谢的有效抑制剂，可能会影响药物的疗效或毒性。意义：作为表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKIs），吉非替尼和厄洛替尼是转移性非小细胞肺癌的一线药物治疗方法。我们的实验结果表明，临床上相关浓度的吉非替尼、去甲氧基丙基吉非替尼、厄洛替尼、去甲基厄洛替尼和二去甲基厄洛替尼，但不是去甲氧基吉非替尼，抑制人醛氧化酶（AOX1）的催化活性和扎来普隆和甲氨蝶呤的肝胞质代谢。分子对接分析提供了对这些 EGFR-TKIs 与 AOX1 关键相互作用的结构见解。我们的研究结果可能会引发对新的 EGFR-TKI 设计和开发的改进策略。
