Multimeric HIV-1 Integrase Inhibitors

摘要 本提案的首要目标是开发一流的多聚体HIV-1整合酶(IN) 用于其未来临床开发的抑制剂(MINI)，并将这些化合物作为强大的研究开发 HIV-1分子生物学工具，用于揭示成熟过程中的关键分子相互作用。因为他们的 独特的作用模式，迷你病毒有望在临床上有效地抑制所有耐药的病毒表型， 它根据当前使用的艺术而不断演变。通过合理修改原型，多功能 基于喹啉的变构IN抑制剂(ALLINI)，我们已经开发出高效的基于吡啶的迷你抑制剂，它 对诱导IN的超多聚化具有很高的选择性。我们的特别行政区研究一直是关键的 了解这些抑制剂的抗病毒作用模式，使我们能够清楚地描述其意义 HIV-1的多聚体作为一种新的、有吸引力的治疗靶点。我们已经展示了这种超多聚化 在成熟过程中，IN发生在病毒颗粒中，这反过来又损害IN与病毒RNA基因组的结合，并 导致具有核糖核蛋白复合体的偏心、非传染性病毒粒子移位到 保护性衣壳核心。此外，我们的合理设计方法使我们能够开发出一种先导化合物MINI KF116，与Allini对应物相比，抗性的遗传屏障明显增强。在……里面 特别是，KF116对具有A128T IN替换的HIV-1变异株具有完全的活性，从而产生抵抗力 对大多数原型艾利尼人来说。相反，三重(T124N/V165I/T174I)IN替换，这显著 即使通过V165I的补偿性突变也会损害病毒的复制，这对于增强对 KF116。总而言之，我们的研究结果表明，基于吡啶的KF116是一个很好的开发平台 第二代迷你车。我们未来的工作将扩展这些研究，并在我们令人振奋的新初步工作的基础上 结果表明，Minis/AlLINI表现出对全长野生型IN四聚体的显著偏好。 具体地说，我们发现与催化核心区(CCD二聚体)结合的抑制剂不足以保证其活性 相邻全长In四聚体之间的Cd-Inhibitor-C-末端结构域相互作用 诱导IN超多聚化所必需的。因此，我们建议剖析独特的结构特征 IN四聚体作为Mini和Allini的真实靶标(目标1)，并利用该信息优化第二 用于其未来临床开发的生成抑制剂(目标2)。拟议的研究具有很高的 补充并将与制药业非常积极的持续努力协同作用，以翻译 一流的迷你/ALLINS进入临床，最终目标是提供更安全的下一代 治疗学。

Abstract The overarching goals of the present proposal are to develop first-in-class multimeric HIV-1 integrase (IN) inhibitors (MINIs) for their future clinical development and to exploit these compounds as powerful investigational tools for HIV-1 molecular biology to uncover critical molecular interactions during maturation. Because of their unique mode of action, MINIs are expected to potently inhibit all drug resistant viral phenotypes in the clinic, which continually evolve in response to currently used ARTs. By rationally modifying archetypal, multifunctional quinoline-based allosteric IN inhibitors (ALLINIs), we have developed highly potent pyridine-based MINIs, which are highly selective for inducing hyper-multimerization of IN. Our SAR studies have been critical for understanding the antiviral mode of action of these inhibitors and allowed us to clearly delineate the significance of HIV-1 IN multimerization as a novel, attractive therapeutic target. We have shown this hyper-multimerization of IN occurs in viral particles during maturation, which in turn impairs IN binding to the viral RNA genome and results in eccentric, non-infectious virions with ribonucleoprotein complexes being displaced outside of the protective capsid core. In addition, our rational design approach enabled us to develop a lead compound, MINI KF116, with a markedly enhanced genetic barrier to resistance compared with its ALLINI counterparts. In particular, KF116 is fully active against the HIV-1 variant with an A128T IN substitution, which confers resistance to the majority of archetypal ALLINIs. Instead, triple (T124N/V165I/T174I) IN substitutions, which significantly compromise viral replication even with a compensatory V165I mutation, are necessary to confer resistance to KF116. Collectively, our findings argue that pyridine-based KF116 is as an excellent platform for the development of second generation MINIs. Our future work will extend these studies and build on our exciting new preliminary results, which show that MINIs/ALLINIs exhibit striking preference for full-length wild type IN tetramers. Specifically, we found that inhibitor binding to the catalytic core domain (CCD) dimer is not sufficient for its activity and that the CCD-inhibitor-C-terminal domain interactions between adjoining full-length IN tetramers are necessary to induce hyper-multimerization of IN. Accordingly, we propose to dissect unique structural features of IN tetramers as authentic targets for MINIs and ALLINIs (aim 1) and utilize this information to optimize second generation inhibitors for their future clinical development (aim 2). The proposed studies are highly complementary to and will synergize with very active ongoing efforts in the pharmaceutical industry to translate the first-in-class MINIs/ALLINIs into the clinic with the ultimate goal of delivering safer next generation therapeutics.