STRUCTURE BASED THERMODYNAMIC STUDIES OF HIV-1 PROTEASE

描述：与治疗相关的最严重的副作用之一 HIV-1感染的一个重要原因是病毒株的出现， 蛋白酶抑制剂。本项目的主要目标是了解 HIV- 1蛋白酶中的特定突变引起 抑制剂耐药性，并将这些知识纳入 药物设计的新策略。一个相关的目标是了解 抑制剂对不同HIV- 1蛋白酶分子的有效性 亚型这些蛋白酶在几个位置含有氨基酸变异， 包括一些与耐药性有关的病毒。主要 该项目将回答的问题是： 1)蛋白酶分子中的特定突变是如何优先降低蛋白酶活性的？ 抑制剂相对于底物的结合亲和力？有哪些 阻力的热力学和结构基础 2)目前的抑制剂对不同HIV- 1蛋白酶的有效性如何 亚型？虽然亚型B在美国占主导地位， 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，并占大多数的艾滋病毒感染 国际吧 3)是热力学的起源，这些力决定了结合亲和力 与耐药突变易感性相关的抑制剂？ 是否有可能鉴别出蛋白酶抑制剂与 焓/熵平衡，但相似的亲和力？他们是否对 以不同的方式产生抗药性突变 4)有没有可能开发出明确的分子设计策略， 考虑到对耐药性的敏感性导致突变？ 这些问题将由实验热力学和 测量（高灵敏度等温滴定量热法和高灵敏度恒温滴定量热法） 灵敏度差示扫描量热法），结构测定（X射线 晶体学）和基于结构的热力学计算。

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.