Effect of Ethanol on the Metabolic Characteristics of HIV-1 Integrase Inhibitor Elvitegravir and Elvitegravir/Cobicistat with CYP3A: An Analysis Using a Newly Developed LC-MS/MS Method.
Effect of Ethanol on the Metabolic Characteristics of HIV-1 Integrase Inhibitor Elvitegravir and Elvitegravir/Cobicistat with CYP3A: An Analysis Using a Newly Developed LC-MS/MS Method.
复制标题
乙醇对 HIV-1 整合酶抑制剂 Elvitegravir 和 Elvitegravir/Cobicistat 与 CYP3A 代谢特征的影响：使用新开发的 LC-MS/MS 方法进行分析
DOI：
10.1371/journal.pone.0149225
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Kumar S
中科院分区：
文献类型：
--
作者：
Midde NM;Rahman MA;Rathi C;Li J;Meibohm B;Li W;Kumar S
Elvitegravir (EVG) 是一种用于治疗 HIV 感染的整合酶抑制剂，正日益成为一线抗逆转录病毒治疗 (ART) 方案的一部分。 EVG主要通过细胞色素P450（CYP）3A4代谢。此前，我们已经证明乙醇会改变 ART-CYP3A4 与蛋白酶抑制剂的相互作用，从而改变它们的代谢。然而，由于 EVG 是一类相当新的药物，其动力学特性以及乙醇对 EVG-CYPP3A4 相互作用的影响知之甚少。在这项研究中，我们表征了 EVG 和考比司他 (COBI) 在人微粒体中促进 EVG 代谢，然后是乙醇-EVG、乙醇-COBI-EVG 与 CYP3A 的相互作用。首先，我们开发并验证了一种简单、灵敏且稳定的液相色谱-串联质谱 (LC-MS/MS) 方法，用于定量人肝微粒体中的 EVG。该药物的定量下限为 0.003 μM (1.34ng/ml)。根据 FDA 指南对所提出方法的提取率、基质效应、药物稳定性和校准曲线进行了验证。时间依赖性动力学数据表明，与单独的 EVG 相比，20mM 乙醇可将 EVG 降解的表观半衰期缩短约 50%。我们的底物动力学结果表明，乙醇轻微降低了 EVG 代谢的催化效率。抑制研究表明，EVG 抑制 CYP3A4，20 mM 乙醇会导致 EVG 的 IC50 降低。然而，在 COBI 存在的情况下，我们无法有效地确定这些参数，因为 COBI 作为 CYP3A4 的强抑制剂，阻断了 EVG/乙醇-CYP3A4 相互作用。对接研究预测，在乙醇存在的情况下，EVG 或 COBI 与 CYP3A4 活性位点的结合会发生变化。综上所述，这些结果表明乙醇与微粒体 CYP3A 相互作用并改变 EVG-CYP3A4 相互作用，从而改变 EVG 代谢和 EVG 对 CYP3A4 的抑制。这一发现具有临床意义，因为饮酒在艾滋病毒人群中非常普遍，并且在这些患者接受抗逆转录病毒治疗期间没有单独的指南。
Elvitegravir (EVG), an integrase inhibitor for the treatment HIV infection, is increasingly becoming the part of first-line antiretroviral therapy (ART) regimen. EVG is mainly metabolized through cytochrome P450 (CYP) 3A4. Previously, we have shown that ethanol alters ART-CYP3A4 interactions with protease inhibitors thereby altering their metabolisms. However, as EVG is a fairly new class of drug, its kinetic characteristics and the effect of ethanol on EVG-CYPP3A4 interaction is poorly understood. In this study, we characterized EVG and cobicistat (COBI)-boosted EVG metabolism in human microsomes followed by ethanol-EVG, ethanol-COBI-EVG interaction with CYP3A. First, we developed and validated a simple, sensitive, and robust liquid chromatography–tandem mass spectrometry (LC-MS/MS) method for the quantification of EVG in the human liver microsomes. The lower limit of quantification for the drug was at 0.003 μM (1.34ng/ml). Extraction yield, matrix effects, drug stability, and calibration curves for the proposed method were validated according to the FDA guidelines. Time dependent kinetics data showed that 20mM ethanol decreases the apparent half-life of EVG degradation by ~50% compared to EVG alone. Our substrate kinetic results revealed that ethanol mildly decreases the catalytic efficiency for EVG metabolism. Inhibition studies demonstrated that EVG inhibits CYP3A4, and 20 mM ethanol causes a decrease in the IC50 of EVG. However, in the presence of COBI we were unable to determine these parameters effectively because COBI, being a strong inhibitor of CYP3A4, blocked the EVG/ethanol-CYP3A4 interactions. Docking studies predicted a shift of EVG or COBI binding to the active site of CYP3A4 in the presence of ethanol. Taken together, these results suggest that ethanol interacts with microsomal CYP3A and alters EVG-CYP3A4 interaction thereby altering EVG metabolism and inhibition of CYP3A4 by EVG. This finding has clinical significance because alcohol use is highly prevalent in HIV population, and there are no separate guidelines for these patients while they are on ART medication.