Ultra-potent HIV capsid inhibitors

超强效 HIV 衣壳抑制剂

• 批准号: 10267756
• 负责人: Francisco J Asturias
• 金额: $ 79.2万
• 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/10267756
• 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.

摘要 抗逆转录病毒疗法(ART)已经将曾经致命的艾滋病毒/艾滋病转变为一种可控的、 慢性感染。然而，与当前艺术相关的一些紧迫问题仍然存在，包括 每天服用HIV-1药物的必要性，治疗依从性不佳，以及 出现耐药病毒表型。因此，有必要发展长效 针对临床未开发的病毒蛋白的抗逆转录病毒药物以缓解上述问题。HIV-1 衣壳蛋白是一个新的、有吸引力的靶点，因为它在病毒的生命周期中扮演着多种重要的角色。 GS-6207(吉利德科学公司的Lenacapavir)是一种新近发现的、一流的、长效和超 有效的HIV-1衣壳蛋白抑制剂。最近完成的一期临床试验(NCT03739866)建议 GS-6207进入2/3期临床试验(NCT04143594/NCT04150068)，为期6个月 给药间隔。我们的研究目标是阐明一种高度有效的 GS-6207的抗病毒活性，并利用所获得的知识开发第二代抑制剂。 为此，我们合成并检测了GS-6207的抗病毒活性。与 衣壳蛋白在HIV-1生物学中的多方面作用，该抑制剂有效地(EC50为~55 PM)损伤传入病毒 并在病毒外流期间表现出第二次略有降低的抗病毒活性(EC50为~314 PM)。模式-- GS-6207的作用研究表明，该抑制剂通过稳定和 从而防止受感染细胞胞质中衣壳的功能性解体。此外, GS-6207干扰衣壳与细胞内HIV-1辅助因子Nup153和CPSF6的结合 病毒核导入和直接整合到染色质的富含基因的区域。我们的X射线结晶学， 低温电子显微镜和氢-氚交换实验表明，GS-6207 紧密结合两个相邻的衣壳亚基，促进远端六聚体内和六聚体之间的相互作用 惊人地稳定了弯曲的衣壳格子。此外，我们的GS-6207的高分辨率X射线结构 结合衣壳六聚体使我们能够在GS-6207附近绘制耐药变体的图谱 结合部位。这些信息将是合理设计第二代抑制剂的关键。我们建议 为了扩大这些研究，以更好地了解GS的多模式、特别有效的抗病毒活性- 6207在HIV-1复制的早期和晚期。为此，我们将追求以下三个方面 具体目标：目标1将阐明抑制艾滋病毒进入后步骤的结构和机制基础- 1感染GS-6207；目标2将剖析抑制病毒产生和 目标3将研究病毒对GS-6207耐药的结构基础 合理开发具有增强抗药性屏障的第二代缓蚀剂。加在一起， 本研究将深入剖析GS-6207的多通道抗病毒作用机制，提供新的抗病毒机制 对衣壳病毒生物学的洞察，确定关键的抑制物-衣壳相互作用并促进优化 这类化合物的临床使用。