Multimeric HIV-1 Integrase Inhibitors

本申请旨在发现多聚体HIV-1整合酶（IN）抑制剂或MINI。HIV-1 IN多聚化呈现重要的治疗靶标，因为有序的多聚体状态对于病毒复制是必需的。我们研究的主要目的是通过稳定而不是破坏相互作用的IN亚基以促进异常蛋白多聚化来损害IN功能。我们最初的概念验证实验为我们的方法的可行性奠定了坚实的基础。我们已经证明，一个小分子可以抑制IN催化活性在体外结合在蛋白质二聚体界面，稳定相互作用的亚基和促进形成更高的订单寡聚体。使用多官能喹啉化合物的最近的研究已经证明，这些化合物有效地靶向感染细胞中的IN多聚化。然而，喹啉化合物作为研究探针的应用是有限的，这是由于它们促进异常IN多聚化并以类似的效力抑制IN-LEDGF/p75结合的事实。因此，需要新的更具选择性的化合物来探索HIV-1 IN多聚化在病毒生命周期中的作用。因此，我们提出了以下三个目标：目标1是合理开发MINI并阐明其抗病毒作用机制;目标2是研究MINI治疗细胞培养中HIV-1引起的IN耐药突变并开发第二代抑制剂;目标3将专注于通过对365，000种化合物的库进行高通量筛选（HTS）以及随后的命中-先导优化来发现新的MINI。拟议的研究将开发新的，强大的研究探针（MINI），通过选择性靶向IN多聚化来研究HIV-1分子生物学。此外，MINI有望靶向HIV-1 IN上以前未利用的位点，并有效抑制对所有目前使用的HAART耐药的HIV-1表型。因此，我们的研究将促进临床上有用的新型变构IN抑制剂的开发。

DESCRIPTION: The present application aims to discover multimeric HIV-1 integrase (IN) inhibitors or MINIs. HIV-1 IN multimerization presents an important therapeutic target as the ordered multimeric state is essential for virus replication. The principal objective of our studiesis to impair IN function by stabilizing rather than destabilizing interacting IN subunits to promote aberrant protein multimerization. Our initial proof-of-concept experiments have laid a strong foundation for the feasibility of our approach. We have demonstrated that a small molecule can inhibit IN catalytic activity in vitro by binding at the protein dimer interface, stabilizing the interacting subunits and promoting formation of higher-order oligomers. More recent studies with multifunctional quinoline compounds have demonstrated that these compounds potently target IN multimerization in infected cells. However, the application of quinoline compounds as investigational probes is limited due to the fact that they promote aberrant IN multimerization and inhibit the IN-LEDGF/p75 binding with similar potency. Therefore, new more selective compounds are needed to explore the roles of HIV-1 IN multimerization in the virus life cycle. Accordingly, we propose the following three aims: Aim 1 will rationally develop MINIs and elucidate their antiviral mechanism of action; Aim 2 will investigate IN resistance mutations that arise from treatment of HIV-1 in cell culture with MINIs and develop second generation inhibitors; Aim 3 will focus on discovery of new MINIs through high-throughput screening (HTS) of a library of 365,000 compounds and subsequent hit-to-lead optimization. The proposed studies will develop novel, powerful investigational probes (MINIs) to study HIV-1 molecular biology by selectively targeting IN multimerization. In addition, MINIs are expected to target previously unexploited sites on HIV-1 IN and potently inhibit HIV-1 phenotypes resistant to all currently used HAART. Thus, our studies will facilitate the development of clinically useful new types of allosteric IN inhibitors.