A New Paradigm for HIV Treatment: Targeted Degradation of HIV Reverse Transcriptase via the Ubiquitin-Proteasome Pathway

HIV 治疗的新范式：通过泛素-蛋白酶体途径靶向降解 HIV 逆转录酶

• 批准号: 10153409
• 负责人: Robert C Reynolds
• 金额: $ 20.54万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-11-06 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10153409
• 关键词: Affinity; Antineoplastic Agents; Antiviral Agents; Binding; Biological Assay; Biological Availability; Biology; CD4 Positive T Lymphocytes; Chronic; Clinic; Clinical; Clinical Treatment; Clinical Trials; Communicable Diseases; Complex; Computing Methodologies; DBL Oncoprotein; Detection; Development; Disease; Drug resistance; Enhancers; Epidemic; Foundations; Future; Genetic Variation; Goals; HIV; HIV Infections; HIV resistance; HIV-1; Hepatitis C virus; Impairment; Infection; Lead; Ligands; Maintenance Therapy; Malignant Neoplasms; Mediating; Modality; Modeling; Molecular Conformation; Mutation; Mycobacterium tuberculosis; Oral; Organic Synthesis; Outcome; Pathway interactions; Patients; Peptide Hydrolases; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacotherapy; Predisposition; Preparation; Prevention; Probability; Property; Proteins; RNA-Directed DNA Polymerase; Reagent; Regimen; Reporting; Request for Applications; Research; Resistance; Resistance development; Reverse Transcriptase Inhibitors; Savings; Series; Site; Structure; Therapeutic; Therapeutic Intervention; Treatment Failure; Treatment outcome; Ubiquitin; Ubiquitination; United States National Institutes of Health; Variant; Viral; Virus Diseases; Virus Replication; analog; anti-cancer; antimicrobial drug; antiretroviral therapy; assay development; base; compliance behavior; design; drug development; drug discovery; experience; fight against; high reward; high risk; in vitro Model; in vivo; inhibitor/antagonist; multicatalytic endopeptidase complex; new technology; non-nucleoside reverse transcriptase inhibitors; novel therapeutic intervention; novel therapeutics; pre-exposure prophylaxis; prevent; programs; protein degradation; protein function; prototype; recruit; resistant strain; screening; simulation; ubiquitin-protein ligase; viral resistance

PROJECT SUMMARY/ABSTRACT In accordance with RFA-AI-19-072, Novel Therapeutics Directed to Intracellular HIV Targets, we propose using targeted protein degradation (TPD) against the essential HIV reverse transcriptase (RT). The RFA states “Controlling protein function by controlling intracellular protein levels has evolved as a promising and novel therapeutic strategy. This can be achieved by the targeted degradation of intracellular proteins thru the ubiquitin- proteasome pathway.” Heterobifunctional targeted protein degraders (TPDs) are being actively pursued as enhancers of proteasomal destruction of proteins specifically associated with several cancers. Such agents offer advantages over traditional occupancy-based inhibitors including a unique catalytic mechanism of action, greater target selectivity, and a reduced probability for resistance development. Surprisingly, this promising therapeutic modality has only recently been applied to antiviral drug discovery through a successful Telapravir-based TPD effective against the wild type and resistant forms of the HCV protease. Analysis of reported HIV-1 drug-bound structures suggests that the essential viral RT should be readily adaptable to targeted protein degradation. The potent and clinically used HIV-1 RT inhibitor (RTI), Rilpivirine (RPV) binds the RT site in Gag-Pol and also allosterically inhibits p66/p51 RT function by binding the non-nucleoside-RTI (NNRTI) Binding Pocket (NNIBP) on the p66 subunit. RPV is amenable to conjugation with linkers and ubiquitin E3 ligase recruiting ligands to serve in the design and preparation of prototype HIV-1 RT TPDs. The OBJECTIVE of this study is to show proof-of-concept of a new inhibitory mechanism by which HIV-1 RT can be targeted for degradation, impairing HIV infectivity and replication. Importantly, due to TPDs’ unique mechanism of action, even a low- affinity RT/TPD interaction will likely lead to effective target degradation. Thus, we pose the HYPOTHESIS that RPV-based TPDs will not only augment inhibition against HIV-1 with RTI-sensitive RT but will remain effective against RTI-resistant RT variants. Ultimately, this approach can reduce resistance development and potentially extend regimen lifetimes in the fight against HIV disease. The objective of AIM 1 is to design and prepare RPV- based TPDs built on state-of-the-art computational methods and predictive physicochemical properties currently accepted for in vivo active TPDs. In AIM 2, we will screen the two series of TPDs for antiviral activity in complementary in vitro models of HIV-1 single-round infection (in the TZM-bl assay), and of replication (in primary CD4+ T lymphocytes). The IMPACT of the discovery of targeted HIV-1 RT degraders that limit infectivity and replication through a mechanism distinct from occupancy-based HIV-1 RTIs will be the identification of agents that are effective against RTI-resistant HIV strains and that limit RTI susceptibility to viral resistance development. Grandly, this research will demonstrate that TPD approaches can be superior in the treatment of HIV disease and will set the stage for the advancement of TPDs against other HIV targets with improvements in the prevention of resistance development and with concomitant enhanced clinical treatment outcomes.

项目摘要/摘要 根据RFA-AI-19-072，针对细胞内HIV靶标的新型治疗剂，我们建议使用 针对基本HIV逆转录酶（RT）的靶向蛋白降解（TPD）。 RFA国家 “通过控制细胞内蛋白水平来控制蛋白质的功能已发展为有望和新颖 治疗策略。这可以通过通过泛素 - 细胞内蛋白的靶向降解来实现 蛋白酶体途径。 与多种癌症特别相关的蛋白质蛋白质蛋白破坏蛋白质的增强子。这样的代理商提供 优于传统占用抑制剂，包括独特的催化作用机制，更大 目标选择性，以及阻力发展的可能性降低。令人惊讶的是，这种承诺疗法 直到最近通过成功的Telapravir TPD将模态应用于抗病毒药物发现 对HCV蛋白酶的野生型和抗性形式有效。分析报告的HIV-1药物结合 结构表明，基本病毒RT应容易适应靶向蛋白质降解。这 潜力和临床使用的HIV-1 RT抑制剂（RTI），Rilpivirine（RPV）结合了GAG-POL中的RT位点，也结合 通过结合非核苷-RTI（NNRTI）结合口袋，变构抑制p66/p51 RT功能 （NNIBP）在p66亚基上。 RPV可以与接头和泛素E3连接酶招募相结合 配体用于HIV-1 RT TPD的设计和制备。这项研究的目的是 显示一种新的抑制机制的概念证明，可以将HIV-1 RT用于降解， 损害艾滋病毒感染和复制。重要的是，由于TPD的独特作用机理，甚至是低 - 亲和力RT/TPD相互作用可能会导致有效的目标降解。那我们提出了这样一个假设 基于RPV的TPD不仅会增加对RTI敏感RT对HIV-1的抑制作用，而且将保持有效 反对耐RT的RT变体。最终，这种方法可以减少抵抗力的发展，并可能 在抗击艾滋病毒疾病的斗争中延长了疗法的寿命。目标1的目的是设计和准备RPV- 基于最先进的计算方法和预测物理属性构建的基于TPD 接受体内活性TPD。在AIM 2中，我们将筛选两个系列的TPD，以进行抗病毒活性 HIV-1单转感染的互补体外模型（在TZM-BL测定中）和复制（在原发性中 CD4+ T淋巴细胞）。发现有针对性的HIV-1 RT降解器的影响，限制了感染和 通过与基于占用率的HIV-1 RTI不同的机制复制将是对代理的识别 可有效防止RTI耐药的HIV菌株，并限制RTI对病毒抗性的敏感性 发展。宏伟的这项研究将表明，TPD方法在治疗中可以是优越的 艾滋病毒疾病，将为TPD朝其他艾滋病毒靶标的发展奠定基础 预防抵抗力发展并随之增强的临床治疗结果。