Physical trapping of HIV-1 synaptic complex by different structural classes of integrase strand transfer inhibitors.
Physical trapping of HIV-1 synaptic complex by different structural classes of integrase strand transfer inhibitors.
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不同结构类别的整合酶链转移抑制剂对 HIV-1 突触复合体的物理捕获。
DOI：
10.1021/bi100514s
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发表时间：
2010
期刊：
影响因子：
2.9
通讯作者：
Grandgenett,DuaneP
中科院分区：
文献类型：
--
作者：
Pandey,KrishanK;Bera,Sibes;Vora,AjaykumarC;Grandgenett,DuaneP
Raltegravir是FDA批准的针对人类免疫缺陷病毒1型（HIV-1）整合酶（IN）的抑制剂。在这项研究中，我们研究了能够抑制HIV-1 In协同整合的多链转移抑制剂的相关机制。结果表明，雷替重力韦、依替重力韦、MK-2048、RDS 1997和RDS 2197似乎都通过改变IN -病毒DNA相互作用而包含一个共同的抑制机制。这些结构不同的抑制剂结合突触复合体并使其失活，这是体外协调整合途径中的一种中间体。抑制剂物理捕获突触复合体，从而阻止目标DNA结合，从而协同整合。在天然琼脂糖凝胶上观察到的特定抑制剂捕获突触复合物的效率与其抑制协同整合反应的效力相关，由每种抑制剂的ic50值定义。在低纳摩尔浓度（<50 nM）下，雷替重力韦显示出协同整合的时间依赖性抑制，这是一种与慢结合抑制剂相关的特性。对无抑制剂的抗雷替重力受体突变体N155H和Q148H的研究表明，它们组装突触复合体和促进协同整合的能力与报道的病毒复制能力相似。与N155H相比，Q148H的协同整合活性显示出更高的对雷替重力韦的交叉抗性，这就解释了为什么继发性突变的Q148H途径在长期治疗中是主要途径。值得注意的是，MK-2048对野生型IN和抗替地韦IN突变体N155H同样有效，这表明该抑制剂可能在它们的药物结合口袋中类似地结合。
Raltegravir is an FDA approved inhibitor directed against human immunodeficiency virus type 1 (HIV-1) integrase (IN). In this study, we investigated the mechanisms associated with multiple strand transfer inhibitors capable of inhibiting concerted integration by HIV-1 IN. The results show raltegravir, elvitegravir, MK-2048, RDS 1997, and RDS 2197 all appear to encompass a common inhibitory mechanism by modifying IN−viral DNA interactions. These structurally different inhibitors bind to and inactivate the synaptic complex, an intermediate in the concerted integration pathway in vitro. The inhibitors physically trap the synaptic complex, thereby preventing target DNA binding and thus concerted integration. The efficiency of a particular inhibitor to trap the synaptic complex observed on native agarose gels correlated with its potency for inhibiting the concerted integration reaction, defined by IC50values for each inhibitor. At low nanomolar concentrations (<50 nM), raltegravir displayed a time-dependent inhibition of concerted integration, a property associated with slow-binding inhibitors. Studies of raltegravir-resistant IN mutants N155H and Q148H without inhibitors demonstrated that their capacity to assemble the synaptic complex and promote concerted integration was similar to their reported virus replication capacities. The concerted integration activity of Q148H showed a higher cross-resistance to raltegravir than observed with N155H, providing evidence as to why the Q148H pathway with secondary mutations is the predominant pathway upon prolonged treatment. Notably, MK-2048 is equally potent against wild-type IN and raltegravir-resistant IN mutant N155H, suggesting this inhibitor may bind similarly within their drug-binding pockets.