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

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，以下简称 HIV）目前感染约 4000 万人在没有治愈方法的情况下，抗逆转录病毒药物的感染人数持续上升。治疗是主要的治疗选择，因为它可以减缓疾病进展并减少新的整合酶 (IN) 链转移抑制剂 (INSTI) 是一类阻断感染的抗逆转录病毒疗法。病毒 DNA 整合到宿主染色体中，这是由病毒 IN 酶介导的过程，在病毒 DNA 末端组装成寡聚核蛋白复合物，称为“INSTI”。选择性靶向整合体并代表临床一线疗法然而，IN变异体的出现。对 INSTI 的耐药性正在成为一个更大的临床问题，结构生物学方法可以揭示这一问题。药物作用和耐药机制，为合理改善药物耐药性提供有用信息当前的 INSTI 与辅助技术相辅相成，例如生化活性测定、生物物理学。热力学和动力学测量、细胞病毒学和多种计算方法，包括自由能计算，该结构精确地详细描述了针对特定 INSTI 的抵抗机制为设计和开发新型第三代 INSTI 来对抗感染提供指导。围绕使用冷冻电子显微镜的革命性进展进行结构研究的方法将显示 INSTI 如何与其天然药物靶标 HIV 嵌体（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以下三个具体目标：(1) 扩大耐药性。并以当前的努力为基础，提供对为什么以及如何选择病毒抗性的机械理解变异体（VRV）是对临床使用的药物多替拉韦（DTG）或最有效的药物的反应而出现的 (2) 广泛确定正在开发的内部化合物 4d，目前正在进行临床前评估；并分析第二类药物治疗产生的耐药性的新机制和途径一代，强调原发突变和药物补偿突变，并提供预测策略 (3)选择对新型第三代治疗产生反应而产生的残留耐药变体基于底物拟态概念合成的 INSTI，其中许多能有效抑制病毒第二代临床使用的 INSTI 药物治疗产生的耐药变异，并解释这项工作将增进我们对新型化合物卓越功效的理解。用于治疗艾滋病毒感染者的一类重要药物，确定感染的机制、途径和模式临床相关的 INSTI 耐药性，并为其合理改进提供具体指南。