STRUCTURE BASED THERMODYNAMIC STUDIES OF HIV-1 PROTEASE

HIV-1 蛋白酶基于结构的热力学研究

• 批准号: 6487551
• 负责人: Ernesto Freire
• 金额: $ 4.29万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-01 至 2005-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6487551
• 关键词: X ray crystallography; antiAIDS agent; aspartic endopeptidases; calorimetry; chemical binding; drug design /synthesis /production; drug resistance; drug screening /evaluation; enzyme activity; gene mutation; genetic strain; human immunodeficiency virus 1; protease inhibitor; protein structure; spectrometry; thermodynamics

DESCRIPTION: One of the most serious side effects associated with the therapy of HIV-1 infection is the appearance of viral strains that exhibit resistance to protease inhibitors. The main goal of this project is to understand the molecular mechanisms by which specific mutations in the HIV- 1 protease elicit inhibitor resistance and to incorporate that knowledge in the development of new strategies for drug design. A related goal is to understand the effectiveness of inhibitors against protease molecules from different HIV- 1 subtypes. These proteases contain amino acid variations at several locations, including some that have been associated with drug resistance. The main questions that this project will answer are: 1) How do specific mutations in the protease molecule preferentially lower the binding affinity of inhibitors relative to the substrate? What are the thermodynamic and structural basis of resistance? 2) How effective are current inhibitors against proteases from different HIV- 1 subtypes? While subtype B is the predominant in the United States and has been the target for drug design, other subtypes are prevalent in Africa, Asia and some European countries, and account for most of the HIV-l infections worldwide. 3) Is the thermodynamic origin of the forces that define the binding affinity of inhibitors related to their susceptibility to resistance-causing mutations? Is it possible to identify protease inhibitors with drastically different enthalpy/entropy balances but similar affinities? Do they respond to resistance-causing mutations in a different way? 4) Is it possible to develop molecular design strategies that explicitly consider the susceptibility to resistance causing mutations? These questions will be answered by a combination of experimental thermodynamic measurements (high sensitivity isothermal titration calorimetry and high sensitivity differential scanning calorimetry), structure determination (x-ray crystallography) and structure-based thermodynamic calculations.

描述：与该疗法相关的最严重的副作用之一 HIV-1感染的一个特征是出现了具有抗药性的病毒株 到蛋白水解酶抑制剂。本项目的主要目标是了解 HIV-1蛋白水解酶特异性突变诱导的分子机制 并将这一知识纳入到开发缓蚀剂 药物设计的新策略。一个相关的目标是理解 抑制剂对不同HIV-1来源的蛋白酶分子的抑制作用 子类型。这些蛋白水解酶在几个位置含有氨基酸变异， 包括一些与抗药性有关的基因。主 本项目将回答的问题包括： 1)蛋白水解酶分子中的特定突变如何优先降低 抑制剂与底物的结合亲和力？什么是 阻力的热力学和结构基础？ 2)目前的抑制剂对不同HIV-1的蛋白水解酶的效果如何 亚型？而B亚型在美国占主导地位，一直是 作为药物设计的目标，其他亚型在非洲、亚洲和 一些欧洲国家，并占了L感染艾滋病毒的大部分 全世界。 3)是定义结合亲和力的力的热力学起源 与它们对引起耐药性的突变的易感性有关的抑制剂？ 有没有可能鉴定出截然不同的蛋白水解酶抑制剂？ 焓/熵平衡，但相似的亲和力？他们有没有回应 以一种不同的方式导致耐药突变？ 4)有没有可能开发出明确的分子设计策略 考虑到对引起突变的耐药性的易感性？ 这些问题将通过实验热力学的组合来回答 测量(高灵敏度等温滴定量热法和高灵敏度 灵敏度差示扫描量热法)、结构测定(x射线 结晶学)和基于结构的热力学计算。