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

项目总结 HIV-1衣壳蛋白(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。在.期间 在之前的资金周期中，我们解决了完整的天然全长六聚体HIV-1CA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倍内六聚体界面上， 影响核心稳定性。我们还解决了包含调节核心稳定性的突变的&gt；45 CA结构，因此 提供了核心稳定的分子基础的第一次一瞥，这是设计潜力的关键 治疗学。其他结构与宿主因子多肽或各种新的抑制剂形成复合体。在……里面 此外，我们合成并表征了针对PF74结合口袋的&gt；220化合物。其中 是比PF74效力高得多的化合物，更重要的是，化合物具有更好的抵抗力 与临床试验中的高效力CA靶向药物GS-6207进行比较。除了强大的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“不稳定剂”，提高了令人兴奋的 可能导致衣壳过早脱壳和干扰素上调的核心不稳定的前景。 我们将结合使用化学合成、病毒学、生物物理学和结晶学方法来 合成一些化合物，通过新的原分子间和原分子内相互作用来提高效力； 克服GS-6207抗药性突变；调节核心稳定性并上调干扰素反应；使 通过创新的机制与CA相互作用，包括共价交联。这些化合物将是 表征和构效关系的研究将以X射线结晶学、生物物理 研究(时间推移成像、热位移、组装动力学分析)和病毒学特征 作用机制以及抗药性研究。这项研究将由斯特凡的团队进行 萨拉菲亚诺斯在埃默里大学(结构，生物物理学，病毒学)和王振全(化学合成，大学 埃里克·弗里德在DRP-NCI(病毒学)工作，并与Greg Melikian(成像)合作。 拟议的研究将加深我们对核心稳定性的结构基础的理解，核心稳定性控制着 病毒生命周期中的大量步骤。它们还将导致确定创新的化合物线索。 具有新的“CA-破坏稳定”作用机制、高效、改进的抗性图谱、CA-交联性 功能性，以及改进的药理特性。

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.