Ultra-potent HIV capsid inhibitors

超强效 HIV 衣壳抑制剂

• 批准号: 10160584
• 负责人: Francisco J Asturias
• 金额: $ 81.98万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-21 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10160584
• 关键词: AIDS/HIV problem; Anti-Retroviral Agents; Antiviral Agents; Binding; Binding Sites; Biochemistry; Biological; Biology; Capsid; Capsid Proteins; Caring; Cells; Chromatin; Clinical; Collaborations; Cryoelectron Microscopy; Cytoplasm; Data; Deuterium; Disease; Distal; Dose; Drug resistance; Exhibits; Generations; Genes; HIV; HIV-1; Hydrogen; Impairment; Infection; Investigation; Knowledge; Life Cycle Stages; Manuscripts; Maps; Mediating; Modeling; Molecular; Nuclear Import; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phase I Clinical Trials; Phase II/III Clinical Trial; Phenotype; Physiological; Play; Production; Proteins; Research; Resistance; Resolution; Roentgen Rays; Role; Science; Structure; Structure-Activity Relationship; Testing; Tube; Update; Variant; Viral; Viral Drug Resistance; Viral Proteins; Virus; Virus Replication; X-Ray Crystallography; antiretroviral therapy; base; chronic infection; cofactor; design; drug resistant virus; experimental study; follow-up; gag Gene Products; improved; inhibitor/antagonist; insight; multimodality; novel; pre-clinical; prevent; targeted agent; therapeutic target; treatment adherence; viral resistance; virology; virus core

Abstract Antiretroviral therapies (ART) have transformed the once deadly HIV/AIDS disease into a manageable, chronic infection. Yet, there are still a number of pressing problems associated with current ARTs, including the necessity of daily administration of HIV-1 medications, suboptimal treatment adherence, and the emergence of drug-resistant viral phenotypes. Therefore, there is a need for developing long-acting antiretroviral agents targeting clinically unexploited viral proteins to mitigate the above problems. HIV-1 capsid protein is a novel, attractive target as its plays multiple essential roles during the virus life cycle. GS-6207 (Lenacapavir, Gilead Sciences) is a recently discovered, first-in-class, long-acting, and ultra- potent HIV-1 capsid inhibitor. Recently completed phase 1 clinical trials (NCT03739866) have suggested advancement of GS-6207 into phase 2/3 clinical trials (NCT04143594/NCT04150068) with a six-month dosing interval. Our research objective is to elucidate structural and mechanistic bases for a highly potent antiviral activity of GS-6207 and exploit the knowledge obtained to develop second-generation inhibitors. For this, we have synthesized and examined the antiviral activities of GS-6207. Consistent with the multifaceted role of capsid in HIV-1 biology, the inhibitor potently (EC50 of ~55 pM) impaired incoming virus and exhibited a second, slightly reduced (EC50 of ~314 pM) antiviral activity during virus egress. Mode-of- action studies of GS-6207 revealed that the inhibitor blocks post-entry steps of infection by stabilizing and thereby preventing functional disassembly of the capsid shell in the cytoplasm of infected cells. In addition, GS-6207 interfered with capsid binding to the cellular HIV-1 cofactors Nup153 and CPSF6 that mediate viral nuclear import and direct integration into gene-rich regions of chromatin. Our x-ray crystallography, cryo-electron microscopy, and hydrogen-deuterium exchange experiments have revealed that GS-6207 tightly binds two adjoining capsid subunits and promotes distal intra- and inter-hexamer interactions that strikingly stabilize the curved capsid lattice. Furthermore, our high-resolution x-ray structure of GS-6207 bound to a capsid hexamer enabled us to map drug-resistant variants in close proximity to the GS-6207 binding site. This information will be critical for rational design of second-generation inhibitors. We propose to extend these studies to better understand the multimodal, exceptionally potent antiviral activity of GS- 6207 during both early and late steps of HIV-1 replication. For this, we will pursue the following three specific aims: Aim 1 will elucidate structural and mechanistic bases for inhibition of post-entry steps of HIV- 1 infection by GS-6207; Aim 2 will dissect underlying mechanisms of inhibition of virus production and maturation by GS-6207; and Aim 3 will investigate the structural basis for viral drug-resistance to GS-6207 and rationally develop second-generation inhibitors with an enhanced barrier to resistance. Taken together, the proposed studies will dissect the multimodal antiviral mechanism of action of GS-6207, provide new insights into the viral biology of capsid, identify key inhibitor-capsid interactions and facilitate optimization of this class of compounds for their clinical use.

抽象的 抗逆转录病毒疗法（ART）已将曾经致命的艾滋病毒/艾滋病疾病转变为可管理的， 慢性感染。但是，仍然存在许多与当前艺术相关的紧迫问题，包括 每日服用HIV-1药物，次优治疗依从性和 耐药病毒表型的出现。因此，需要发展长效 靶向临床未开发的病毒蛋白来减轻上述问题的抗逆转录病毒剂。 HIV-1 衣壳蛋白是一种新颖，有吸引力的靶标，因为它在病毒生命周期中起了多种基本作用。 GS-6207（Lenacapavir，Gilead Sciences）是最近发现的，第一类，长效和超级 - 有效的HIV-1衣壳抑制剂。最近完成的1期临床试验（NCT03739866）已提出 将GS-6207的进步分为2/3阶段临床试验（NCT04143594/NCT04150068） 给药间隔。我们的研究目标是阐明高度有效的结构和机械基础 GS-6207的抗病毒活性并利用获得的知识来发展第二代抑制剂。 为此，我们合成并检查了GS-6207的抗病毒活性。与 衣壳在HIV-1生物学中的多方面作用，抑制剂有效（EC50，约55 pm）受损传入病毒 并在病毒出口期间表现出第二次，略微降低（EC50约314 pm）。模式 GS-6207的行动研究表明，抑制剂通过稳定和稳定感染后的抑制剂阻止 从而防止受感染细胞的细胞质中衣壳壳的功能拆卸。此外， GS-6207干扰了与细胞HIV-1辅助因子NUP153和CPSF6介导的衣壳结合的CPSID结合 病毒核进口和直接整合到富基因的染色质区域。我们的X射线晶体学， 冷冻电子显微镜和氢 - 居民交换实验表明GS-6207 紧密结合两个毗邻的衣壳亚基，并促进远端内部和杂物间相互作用 惊人的稳定弯曲的衣壳晶格。此外，我们的GS-6207的高分辨率X射线结构 绑定到Capsid Hexamer，使我们能够在GS-6207附近绘制耐药的变体 绑定位点。该信息对于第二代抑制剂的合理设计至关重要。我们建议 扩展这些研究以更好地了解GS-的多模式，异常有效的抗病毒活性 6207在HIV-1复制的早期和晚期中。为此，我们将追求以下三个 具体目的：AIM 1将阐明结构和机械基础，以抑制HIV的进入后步骤 GS-6207感染1； AIM 2将剖析抑制病毒产生的潜在机制， GS-6207的成熟； AIM 3将研究GS-6207病毒药物抗药性的结构基础 并在合理地发展了第二代抑制剂，具有增强的阻力障碍。在一起， 拟议的研究将剖析GS-6207的多模式抗病毒作用机理，提供新的 洞悉CAPSID的病毒生物学，识别关键的抑制剂capsid相互作用并促进优化 这类化合物用于临床用途。