Effect of Methamphetamine on Spectral Binding, Ligand Docking and Metabolism of Anti-HIV Drugs with CYP3A4.
Effect of Methamphetamine on Spectral Binding, Ligand Docking and Metabolism of Anti-HIV Drugs with CYP3A4.
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DOI：
10.1371/journal.pone.0146529
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Kumar A
中科院分区：
文献类型：
--
作者：
Nookala AR;Li J;Ande A;Wang L;Vaidya NK;Li W;Kumar S;Kumar A
细胞色素P450 3A4 （CYP3A4）是主要的药物代谢酶，参与抗逆转录病毒药物尤其是蛋白酶抑制剂（PIs）的代谢。本研究旨在研究甲基苯丙胺对PIs与CYP3A4结合和代谢的影响。结果表明，甲基苯丙胺与CYP3A4结合后呈现I型光谱变化，δAmax和KD分别为0.016±0.001和204±18 μM。甲基苯丙胺-CYP3A4对接表明，甲基苯丙胺以两种模式与CYP3A4的血红素结合，均导致n -去甲基化。然后我们研究了甲基苯丙胺与CYP3A4结合对pi的影响。我们的研究结果表明，甲基苯丙胺改变奈非那韦的光谱结合，但对其他I型pi（洛匹那韦、阿扎那韦、替普那韦）没有影响。奈非那韦在δAmax（0.004±0.0003 vs. 0.0068±0.0001）和KD（1.42±0.36 vs.2.93±0.08 μM）水平下的光谱结合变化。我们进一步测试了甲基苯丙胺对2种II型pi结合的影响；利托那韦和茚地那韦。结果表明，甲基苯丙胺通过降低δAmax（0.0038±0.0003比0.0055±0.0003）和KD（0.043±0.0001比0.065±0.001 nM）改变了利托那韦与CYP3A4的结合，而茚地那韦只降低了KD（0.086±0.01比0.174±0.03 nM）。此外，在高CYP3A4水平的人肝微粒体中，LC-MS/MS研究显示，在甲基苯丙胺的存在下，羟基利托那韦的形成减少。最后，CYP3A4与洛匹那韦和利托那韦在甲基苯丙胺不存在和不存在的情况下的对接表明，甲基苯丙胺改变了利托那韦的对接，这与光谱结合和代谢研究的结果一致。总的来说，我们的结果证明了甲基苯丙胺对PIs与CYP3A4结合和代谢的不同影响。这些发现在抗逆转录病毒药物剂量调整方面具有临床意义，特别是在滥用甲基苯丙胺的hiv -1感染个体中使用利托那韦增强抗逆转录病毒治疗。
Cytochrome P450 3A4 (CYP3A4) is the major drug metabolic enzyme, and is involved in the metabolism of antiretroviral drugs, especially protease inhibitors (PIs). This study was undertaken to examine the effect of methamphetamine on the binding and metabolism of PIs with CYP3A4. We showed that methamphetamine exhibits a type I spectral change upon binding to CYP3A4 with δAmax and KD of 0.016±0.001 and 204±18 μM, respectively. Methamphetamine-CYP3A4 docking showed that methamphetamine binds to the heme of CYP3A4 in two modes, both leading to N-demethylation. We then studied the effect of methamphetamine binding on PIs with CYP3A4. Our results showed that methamphetamine alters spectral binding of nelfinavir but not the other type I PIs (lopinavir, atazanavir, tipranavir). The change in spectral binding for nelfinavir was observed at both δAmax (0.004±0.0003 vs. 0.0068±0.0001) and KD (1.42±0.36 vs.2.93±0.08 μM) levels. We further tested effect of methamphetamine on binding of 2 type II PIs; ritonavir and indinavir. Our results showed that methamphetamine alters the ritonavir binding to CYP3A4 by decreasing both the δAmax (0.0038±0.0003 vs. 0.0055±0.0003) and KD (0.043±0.0001 vs. 0.065±0.001 nM), while indinavir showed only reduced KD in presence of methamphetamine (0.086±0.01 vs. 0.174±0.03 nM). Furthermore, LC-MS/MS studies in high CYP3A4 human liver microsomes showed a decrease in the formation of hydroxy ritonavir in the presence of methamphetamine. Finally, CYP3A4 docking with lopinavir and ritonavir in the absence and presence of methamphetamine showed that methamphetamine alters the docking of ritonavir, which is consistent with the results obtained from spectral binding and metabolism studies. Overall, our results demonstrated differential effects of methamphetamine on the binding and metabolism of PIs with CYP3A4. These findings have clinical implication in terms of drug dose adjustment of antiretroviral medication, especially with ritonavir-boosted antiretroviral therapy, in HIV-1-infected individuals who abuse methamphetamine.