STRUCTURE BASED THERMODYNAMIC STUDIES OF HIV-1 PROTEASE

HIV-1 蛋白酶基于结构的热力学研究

基本信息

批准号：
6636242
负责人：
Ernesto Freire
金额：
$ 32.98万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
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/6636242
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

项目摘要

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：与治疗相关的最严重的副作用之一 HIV-1 感染的主要原因是出现了具有抗药性的病毒株至蛋白酶抑制剂。该项目的主要目标是了解 HIV-1蛋白酶的特定突变引起的分子机制抑制剂耐药性并将这些知识纳入开发药物设计的新策略。一个相关的目标是了解不同 HIV-1 蛋白酶分子抑制剂的有效性亚型。这些蛋白酶在多个位置含有氨基酸变异，包括一些与耐药性相关的因素。主要该项目将回答的问题是： 1）蛋白酶分子中的特定突变如何优先降低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) 目前针对不同 HIV-1 蛋白酶的抑制剂效果如何亚型？虽然 B 亚型在美国占主导地位，并且已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，占 HIV-1 感染的大部分全世界。 3) 是定义结合亲和力的力的热力学起源抑制剂与它们对引起耐药性的突变的易感性有关吗？是否有可能鉴定出具有显着不同的蛋白酶抑制剂？焓/熵平衡但相似的亲和力？他们是否回应以不同的方式引起耐药性突变？ 4) 是否有可能开发明确的分子设计策略考虑对引起突变的耐药性的敏感性吗？这些问题将通过实验热力学的结合来回答测量（高灵敏度等温滴定量热法和高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。