New Inhibitors Targeting HIV-1 Integrase During Viral Maturation

摘要 HIV-1整合酶(IN)是被证实的临床靶点，对病毒复制是必不可少的。在早期 在感染步骤中，四聚体IN与病毒DNA的两个末端结合，并催化它们整合到 宿主染色体。据报道，一类新的变构蛋白抑制剂(ALLINI)可以抑制酶 通过促进多聚体中的异常发挥作用。有趣的是，在受感染的细胞中，这些化合物 削弱了HIV-1复制的早期和晚期。特别是，经Allini处理的病毒粒子 发现病毒核心形态存在严重缺陷，与几个 突变体中的复制缺陷II类。总而言之，这些研究表明，除了 其已知的催化功能在早期阶段，IN也发挥着必不可少的非催化作用。 HIV-1复制的后期阶段。我们最近发现IN与几种病毒RNA结合 (VRNA)元素，ALLIN强烈调节病毒粒子中的这些相互作用。此外，我们 观察到IN可以将这些结合的vRNA元件聚集在一起，这表明它可能在vRNA中发挥作用 冷凝。本申请将检验以下假设：In-vRNA相互作用和IN 齐聚作用对于感染性病毒粒子的正确形成是至关重要的，可以通过 小分子抑制剂。在AIMS 1中，我们将充分表征IN-vRNA复合体，并阐明 适当的IN多聚体对vRNA结合和颗粒成熟的意义。在AIMS 2中，我们 将设计新的IN抑制剂，能够调节IN-vRNA结合和颗粒成熟。 从这些研究中发现的机制和分子细节将指导未来的药物靶向 在病毒复制的后期，这些关键的非催化IN功能的启动。

Abstract HIV-1 integrase (IN) is validated clinical target and is essential for viral replication. During the early steps of infection, tetrameric IN associates with two viral DNA ends and catalyzes their integration into the host chromosome. A new class of allosteric IN inhibitors (ALLINIs) has been reported to inhibit enzyme function by promoting aberrant IN multimerization. Interestingly, in infected cells these compounds impaired both early and late stages of HIV-1 replication. In particular, the ALLINI-treated virions were found to display profound viral core morphologies defects similar to those observed with several replication-defective class II IN mutants. Collectively, these studies have suggested that in addition to its known catalytic function in early stage, IN also plays an essential and non-catalytic role during the late stage of HIV-1 replication. We were recently able to discover that IN binds to several viral RNA (vRNA) elements and that ALLINIs strongly modulate these interactions in the virion. In addition, we observed that IN can bring these bound vRNA elements together suggesting a possible role in vRNA condensation. The present application will test the following hypothesis: IN-vRNA interactions and IN oligomerization which are both critical for the correct formation of infectious virions can be targeted by small molecule inhibitors. In aims 1, we will fully characterize the IN-vRNA complex and elucidate the significance of the proper IN multimerization for vRNA binding and particle maturation. In aims 2, we will design new IN inhibitors capable of modulating IN-vRNA binding and particle maturation. Mechanistic and molecular details that will emerge from these studies will guide future drug targeting initiatives of these critical non-catalytic IN functions in late stage of the viral replication.