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

基本信息

批准号：
10551720
负责人：
Dmitry Lyumkis
金额：
$ 68.65万
依托单位：
SALK INSTITUTE FOR BIOLOGICAL STUDIES
依托单位国家：
美国
项目类别：
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/10551720
关键词：
AIDS/HIV problem 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 Individual Infection Integrase Kinetics Lead Light Measurement Mediating Methods Mutation Naphthyridines Nucleoproteins Pathway interactions Patient-Focused Outcomes Patients Pattern Persons Pharmaceutical Preparations Pharmacologic Substance Positioning Attribute Prevention Process 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

AIDS/HIV problem 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 Individual Infection Integrase Kinetics Lead Light Measurement Mediating Methods Mutation Naphthyridines Nucleoproteins Pathway interactions Patient-Focused Outcomes Patients Pattern Persons Pharmaceutical Preparations Pharmacologic Substance Positioning Attribute Prevention Process 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

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项目摘要

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.

抽象的人类免疫缺陷病毒1型（HIV-1，以下称为HIV）当前感染了约4000万全球人民，受感染者的数量继续增加。在没有治疗的情况下，抗逆转录病毒治疗代表了主要治疗选择，因为它减慢了疾病的进展并减少了新的治疗方法整合酶（IN）链转移抑制剂（Instis）是一类抗逆转录病毒疗法病毒DNA整合到宿主染色体中，该过程是由酶中病毒介导的，该过程是在病毒DNA的末端组装成寡聚核蛋白复合物，称为“ intoses”。学院有选择地靶向静态并代表诊所中的一线疗法。但是，变体的出现对Instis的抗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抗药性的基础机制，提供有用的信息以合理改善当前学院。使用辅助技术（例如生化活性测定）完成时，生物物理热力学和动力学测量，细胞病毒学以及潜水的计算方法，包括自由能计算，结构精确地详细介绍了针对特定研究所的抵抗机制和为设计和开发新颖的第三代研究提供了指导，以抗击感染。在此提案中，围绕使用革命性进步进行结构研究的革命进步的方法将显示Intist与其自然药物靶标的HIV Intasome（WT和突变体）的相互作用，并阐明出现对这些药物的抗性的机制。有三个具体目标将：（1）扩展并基于目前的努力，以提供对为什么和如何选择耐药性的机械理解响应于临床使用的药物DoluteGravir（DTG）或最有效的变体（VRV）出现 4D的发育内部化合物，目前正在临床前评估中；（2）广泛识别并分析新的机制和耐药性途径，这些机制是响应第二种治疗而产生的一代药物，强调主要和补偿性突变，并提供预测的策略未来变体；（3）选择针对新的第三代处理而产生的残留抗性变体根据底物模仿的概念合成的研究所，其中许多有效抑制病毒响应第二代临床使用的Insti药物治疗而产生的抗性变体，并解释新型化合物效力的基础机制。这项工作将提高我们对一类重要的药物，用于治疗艾滋病毒感染者，识别机制，途径和模式临床上相关的抗药性，并为其合理的改进提供了具体指南。