Multimeric HIV-1 Integrase Inhibitors

描述：本申请旨在发现多聚体HIV-1整合酶(IN)抑制剂或MinI。HIV-1IN多聚体是一个重要的治疗靶点，因为有序的多聚体状态对病毒复制是必不可少的。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亚基来促进异常蛋白的多聚体来损害IN的功能。我们最初的概念验证实验为我们方法的可行性奠定了坚实的基础。我们已经证明，在体外，一个小分子可以通过结合在蛋白质二聚体界面上，稳定相互作用的亚基，促进更高阶低聚物的形成来抑制IN的催化活性。最近对多功能喹啉化合物的研究表明，这些化合物有效地靶向感染细胞中的IN多聚体。然而，喹啉类化合物作为研究探针的应用受到限制，因为它们促进IN的异常多聚化，并以相似的效力抑制IN-LEDGF/p75的结合。因此，需要新的、更有选择性的化合物来探索HIV-1在病毒生命周期中的多聚体作用。因此，我们提出了以下三个目标：目标1将合理开发迷你基因并阐明其抗病毒作用机制；目标2将研究在细胞培养中用迷你基因处理HIV-1所产生的IN耐药突变，并开发第二代抑制剂；目标3将专注于通过高通量筛选(HTS)365,000化合物文库和随后的点击到领先的优化发现新的迷你基因。拟议的研究将开发新的、强大的调查探针(MINI)，通过选择性地靶向IN多聚体来研究HIV-1分子生物学。此外，迷你病毒有望针对HIV-1IN上以前未被开发的部位，并有效地抑制对目前使用的所有HAART具有耐药性的HIV-1表型。因此，我们的研究将有助于开发临床上有用的新型变构IN抑制剂。

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.