Allosteric integrase inhibitor potency is determined through the inhibition of HIV-1 particle maturation
Allosteric integrase inhibitor potency is determined through the inhibition of HIV-1 particle maturation
复制标题
DOI：
10.1073/pnas.1300703110
复制
发表时间：
2013-05-21
影响因子：
11.1
通讯作者：
Engelman, Alan
中科院分区：
文献类型：
--
作者：
Jurado, Kellie A.;Wang, Hao;Engelman, Alan
整合对于HIV-1复制是必不可少的，并且病毒整合酶（IN）蛋白是重要的治疗靶标。在宿主蛋白透镜上皮衍生生长因子（LEDGF）/转录辅激活因子p75的结合位点处接合IN二聚体界面的变构IN抑制剂（ALLINI）是一类新兴的小分子拮抗剂。与多价药物靶标的抑制一致，ALLINI显示出离体陡峭的抗病毒剂量-反应曲线。ALLINI在体外使IN蛋白多聚化并一致地阻断其与病毒DNA的组装，表明两种整合相关功能（IN催化和IN-LEDGF/p75相互作用）的破坏决定了ALLINI作用的多模式机制。我们现在证明，ALLINI效力是出乎意料地占在HIV-1复制的晚期阶段。所述化合物促进病毒粒子IN多聚化，并使人联想到II类IN突变，阻断电子致密病毒核心的形成，并抑制逆转录和在随后感染的靶细胞中的整合。成熟的病毒粒子对ALLINI处理是不敏感的，并且在病毒产生期间化合物效力不依赖于LEDGF/p75表达水平。我们的结论是，ALLINI的IN的合作多聚化以及LEDGF/p75在病毒从细胞中排出期间不能有效地与病毒接合强调了ALLINI作用的多模式机制。我们的研究结果突出了变构抑制剂的多功能性，主要是抑制病毒复制的一个步骤，这是不同的催化要求的目标酶。IN在HIV-1复制晚期对小分子的脆弱性揭示了在临床ALLINI开发中开发的药理学阿喀琉斯之踵。
Integration is essential for HIV-1 replication, and the viral integrase (IN) protein is an important therapeutic target. Allosteric IN inhibitors (ALLINIs) that engage the IN dimer interface at the binding site for the host protein lens epithelium-derived growth factor (LEDGF)/transcriptional coactivator p75 are an emerging class of small molecule antagonists. Consistent with the inhibition of a multivalent drug target, ALLINIs display steep antiviral dose-response curves ex vivo. ALLINIs multimerize IN protein and concordantly block its assembly with viral DNA in vitro, indicating that the disruption of two integration-associated functions, IN catalysis and the IN-LEDGF/p75 interaction, determines the multimode mechanism of ALLINI action. We now demonstrate that ALLINI potency is unexpectedly accounted for during the late phase of HIV-1 replication. The compounds promote virion IN multimerization and, reminiscent of class II IN mutations, block the formation of the electron-dense viral core and inhibit reverse transcription and integration in subsequently infected target cells. Mature virions are recalcitrant to ALLINI treatment, and compound potency during virus production is independent of the level of LEDGF/p75 expression. We conclude that cooperative multimerization of IN by ALLINIs together with the inability for LEDGF/p75 to effectively engage the virus during its egress from cells underscores the multimodal mechanism of ALLINI action. Our results highlight the versatile nature of allosteric inhibitors to primarily inhibit viral replication at a step that is distinct from the catalytic requirement for the target enzyme. The vulnerability of IN to small molecules during the late phase of HIV-1 replication unveils a pharmacological Achilles' heel for exploitation in clinical ALLINI development.