Structural basis for activity of and resistance to HIV integrase inhibitors

HIV整合酶抑制剂的活性和耐药性的结构基础

• 批准号: 10661078
• 负责人: Dmitry Lyumkis
• 金额: $ 67.19万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-25 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10661078
• 关键词: Active Sites; Address; Affect; Anti-Retroviral Agents; Appearance; Binding; Biochemical; Biological Assay; Biophysics; Cell Culture Techniques; Chromatin; Chromosomes; Clinic; Clinical; Clinical Pathways; Collaborations; Communities; Complement; Complex; Computer Models; Computing Methodologies; Cryoelectron Microscopy; DNA; Data; Development; Disease Progression; Drug Targeting; Drug resistance; Drug resistance pathway; Drug usage; Enzymes; Foundations; Free Energy; Funding; Future; Generations; Guidelines; HIV; HIV Integrase; HIV Integrase Inhibitors; HIV drug resistance; HIV therapy; HIV-1; HIV/AIDS; Individual; Infection; Integrase; Kinetics; Lead; Measurement; Mediating; Methods; Mutation; Naphthyridines; Nucleoproteins; Pathway interactions; Patient-Focused Outcomes; Patients; Pattern; Persons; Pharmaceutical Preparations; Pharmacologic Substance; Positioning Attribute; Prevention; Process; Productivity; Recording of previous events; Residual state; Resistance; Resistance development; Series; Spumavirus; Structure; Techniques; Therapeutic; Therapeutic Uses; Thermodynamics; Variant; Viral; Virus Integration; Virus Replication; Work; analog; antiretroviral therapy; base; clinically relevant; design; drug action; effective therapy; experimental study; fighting; human pathogen; improved; inhibitor; inhibitor therapy; insight; interest; mimicry; mutant; novel; pre-clinical; preclinical evaluation; prototype; public health relevance; resistance mechanism; response; structural biology; tool; treatment response; viral DNA; viral resistance; virology

ABSTRACT The Human Immunodeficiency Virus Type 1 (HIV-1, hereafter referred to as HIV) currently infects ~40 million people worldwide, and the number of infected individuals continues to rise. In the absence of a cure, antiretroviral therapy represents the primary treatment option, because it slows disease progression and reduces new infections. Integrase (IN) Strand Transfer Inhibitors (INSTIs) are a class of antiretroviral therapeutics that block integration of viral DNA into host chromosomes, a process that is mediated by the viral IN enzyme, which assembles into oligomeric nucleoprotein complexes on the ends of viral DNA, termed “intasomes”. INSTIs selectively target intasomes and represent first-line therapies in the clinic. However, the emergence of IN variants resistant to INSTIs is becoming a greater clinical problem. Structural biology approaches can shed light on the mechanisms underlying drug action and resistance, providing useful information for rationally improving the current INSTIs. When complemented with ancillary techniques, such as biochemical activity assays, biophysical thermodynamic and kinetic measurements, cellular virology, and diverse computational approaches including free energy calculations, the structures precisely detail mechanisms of resistance against specific INSTIs and provide guidance for designing and developing novel 3rd generation INSTIs to fight infections. In this proposal, approaches centered around using revolutionary advances in cryo-electron microscopy for structural studies will show how INSTIs interact with their natural drug target, the HIV intasome (both WT and mutant), and elucidate the mechanisms by which resistance to these drugs emerges. There are three Specific Aims that will: (1) extend and build upon current efforts to provide a mechanistic understanding of both why and how select viral resistant variants (VRVs) arise in response to the clinically used drug Dolutegravir (DTG) or the most potent developmental in-house compound that 4d, which is currently under pre-clinical evaluation; (2) broadly identify and analyze novel mechanisms and pathways of drug resistance that arise in response to treatment with 2nd generation drugs, highlighting both primary and compensatory mutations, and providing strategies to predict future variants; (3) select for residual resistant variants arising in response to treatment with novel 3rd generation INSTIs that were synthesized based on the concept of substrate mimicry, many of which effectively inhibit viral resistant variants that arise in response to treatment with 2nd generation clinically used INSTI drugs, and explain mechanisms underlying the superior potency of novel compounds. This work will improve our understanding of an important class of drugs used to treat people living with HIV, identify mechanisms, pathways, and patterns of clinically relevant resistance to INSTIs, and provide specific guidelines for their rational improvement.

摘要 人类免疫缺陷病毒1型（HIV-1，以下简称HIV）目前感染约4000万人， 世界各地的人，受感染的人数继续上升。在没有治愈方法的情况下， 治疗是主要的治疗选择，因为它减缓了疾病进展并减少了新的 感染.整合酶（IN）链转移抑制剂（INSTI）是一类阻断逆转录病毒的抗逆转录病毒治疗药物。 病毒DNA整合到宿主染色体中，这一过程由病毒IN酶介导， 在病毒DNA末端组装成寡聚核蛋白复合物，称为“整合体”。INSTI 选择性靶向intasomes并代表临床一线治疗。然而，IN变体的出现 对INSTIs的耐药性正在成为更大的临床问题。结构生物学方法可以阐明 药物作用和耐药性的机制，为合理改善药物作用和耐药性提供有用的信息。 目前的INSTI。如果辅以辅助技术，如生物化学活性测定、生物物理 热力学和动力学测量，细胞病毒学，以及各种计算方法，包括 自由能计算，结构精确地详细说明了对特定INSTI的抗性机制， 为设计和开发新型第三代INSTI以对抗感染提供指导。在这一提议中， 以利用冷冻电子显微镜的革命性进展进行结构研究为中心的方法将 显示INSTI如何与其天然药物靶点HIV整合体（WT和突变体）相互作用，并阐明 对这些药物产生耐药性的机制。有三个具体目标：（1）扩大 并建立在目前的努力，以提供一个机械的理解，为什么和如何选择病毒抵抗 VRV变异体（VRV）的出现是对临床使用的药物Dolutegravir（DTG）或最有效的 开发内部化合物4d，目前正在进行临床前评估;（2）广泛识别 并分析新的机制和途径的耐药性，出现在应对治疗与第二代 代药物，突出原发性和代偿性突变，并提供预测 未来的变体;（3）选择响应于新的第3代治疗而产生的残留耐药变体 基于底物模拟概念合成的INSTI，其中许多有效抑制病毒， 对第二代临床使用的CIMI药物治疗产生的耐药变异，并解释 新化合物的上级效能的潜在机制。这项工作将提高我们对 这是一类重要的药物，用于治疗艾滋病毒感染者，确定机制，途径和模式， 临床相关的耐药性INSTI，并提供其合理的改善具体的指导方针。