Development of HIV capsid-targeting antivirals that affect immune response by modulating capsid stability and have improved resistance profiles

项目摘要 HIV-1衣壳（CA）是一种在病毒生命周期的多个步骤中发挥重要作用的蛋白质。期间 上一个资金周期，我们解决了完整的天然全长六聚体HIV-1 CA的难以捉摸的结构 揭示了以前未知的分子细节，特别是在3倍和2倍六聚体内界面， 影响核心稳定性。我们还解决了>45个CA结构，包括调节核心稳定性的突变，因此 提供了第一次瞥见核心稳定的分子基础，这对设计潜力至关重要。 治疗学其他结构与宿主因子肽或各种新型抑制剂复合。在 此外，我们合成并表征了超过220种靶向PF 74结合口袋的化合物。其中 是比PF 74效力高得多的化合物，重要的是， GS-6207是一种在临床试验中的高效CA靶向药物。除了强CA六聚体 稳定剂，我们引入了一类创新的化合物，CA六聚体的“去稳定剂”，提高了令人兴奋的 核心不稳定的前景可能导致过早的衣壳脱壳和干扰素上调。 我们将结合化学合成、病毒学、生物物理学和晶体学方法， 合成许多通过新的原聚体间和原聚体内相互作用提高效力的化合物; 克服GS-6207抗性突变;调节核心稳定性和上调干扰素应答;使 通过创新的机制，包括共价交联与CA的相互作用。述化合物将 表征和结构活性关系的研究将由X射线晶体学，生物物理学， 研究（延时成像，热位移，组装动力学测定）和病毒学表征 作用机制以及耐药性研究。这项研究将由斯特凡的小组进行， Sarafianos在埃默里大学（结构，生物物理学，病毒学）和ZQ Wang（化学合成， 明尼苏达州），DRP-NCI的Eric Freed（病毒学），以及与Greg Melikian（成像）的合作。 拟议的研究将促进我们对核心稳定性的结构基础的理解，核心稳定性控制着一个 病毒生命周期中的许多步骤。他们也将导致创新化合物铅的识别 具有新的“CA-去稳定化”作用机制，高效力，改善的抗性特征，CA-交联 功能性和改善的药理学性质。

PROJECT SUMMARY The HIV-1 capsid (CA) is a protein that plays a major role in multiple steps of the virus life cycle. During the previous funding cycle we solved the elusive structure of the intact native full length hexameric HIV-1 CA revealing previously unknown molecular details, especially at the 3-fold and 2-fold intra-hexamer interfaces that affect core stability. We also solved >45 CA structures that include mutations that modulate core stability, thus providing the first glimpses of the molecular basis of core stabilization that is critical in designing potential therapeutics. Additional structures were in complex with host factor peptides, or a variety of novel inhibitors. In addition, we synthesized and characterized >220 compounds that target the PF74-binding pocket. Among them are compounds with much higher potency than PF74, and importantly, compounds with improved resistance profile compared to GS-6207, a highly potent CA-targeting drug in clinical trials. In addition to strong CA hexamer stabilizers, we introduced an innovative class of compounds, “destabilizers” of CA hexamers, raising the exciting prospect of core destabilization that may lead to premature capsid uncoating and interferon upregulation. We will use a combination of chemical synthesis, virological, biophysical, and crystallography approaches to synthesize a number of compounds that improve potency through new inter- and intra-protomer interactions; overcome GS-6207 resistance mutations; modulate core stability and upregulate interferon response; enable interactions with CA through innovative mechanisms, including covalent crosslinking. The compounds will be characterized and the structure activity relationship studies will be guided by X-ray crystallography, biophysical studies (time-lapse imaging, thermal shift, assembly kinetics assays) and virological characterization of mechanism of action as well as drug resistance studies. The research will be conducted by the groups of Stefan Sarafianos at Emory Univ (structure, biophysics, virology) and ZQ Wang (chemical synthesis, Univ of Minnesota), Eric Freed at DRP-NCI (virology), and in collaboration with Greg Melikian (imaging). The proposed studies will advance our understanding of the structural basis of core stability, which controls a large number of steps in the virus life cycle. They will also lead to the identification of innovative compound leads with novel “CA-destabilizing” mechanisms of action, high potency, improved resistance profiles, CA-crosslinking functionalities, and improved pharmacological properties.