STRUCTURE-FUNCTION STUDY OF HIV PROTEASE DRUG RESISTANCE

HIV蛋白酶耐药性的结构功能研究

• 批准号: 6526035
• 负责人: LADISLAU Christopher KOVARI
• 金额: $ 14.42万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-08-01 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6526035
• 关键词: X ray crystallography; active sites; aspartic endopeptidases; drug resistance; enzyme activity; enzyme structure; enzyme substrate; gene mutation; human immunodeficiency virus 1; ligands; molecular dynamics; protease inhibitor; virus protein

The long-term goal of this project is to investigate the structural basis of HIV protease drug resistance and to use that knowledge to devise strategies to overcome resistance. The novel hypothesis being tested is that HIV-1 protease drug resistance is caused by mutations that expand the active site of the protease and as a result the inhibitors bind with lower affinity to the variant forms of the enzyme. The short-term goal of this project is to study the structure and function of four multiple drug resistant HIV protease clinical isolates by using a combined structural, virological and biochemical approach. To accomplish this we will: Aim 1. Analyze the structures of inhibitor complexes of four multiple drug resistant "signature mutants" (807, 3761, 1385, 769) by using X-ray crystallographic and molecular modeling techniques. 30 modeled structures of protease variant-ligand complexes will be systematically compared to determine changes in inhibitor or substrate binding. One-third of the modeled structures will be verified by X-ray crystallography. Aim 2. Measure the virological (IC50) and enzymatic (Ki, Km, Vmax) indicators of drug resistance for a series of four multiple drug resistant HIV- 1 proteases (807, 3761, 1385, 769) and correlate the functional results with structural information derived from Aim 1. The structure-function data from the four multiple drug resistant clinical isolates will be used to examine drug resistance hypotheses including the novel HIV protease active site expansion hypothesis. If the active site expansion hypothesis is confirmed, it will suggest a logical approach to overcome drug resistance. Solving the structural problem of why some mutant HIV-1 proteases cannot be inhibited by the standard inhibitors and using this information to design new and effective compounds is of great importance to human welfare.

该项目的长期目标是研究HIV蛋白酶耐药性的结构性基础，并利用这些知识来制定策略来克服抵抗力。正在测试的新型假设是，HIV-1蛋白酶耐药性是由扩大蛋白酶活性位点的突变引起的，因此抑制剂与较低亲和力与酶变体形式结合。该项目的短期目标是通过使用联合结构，病毒学和生化方法研究四个多种耐药性HIV蛋白酶临床分离株的结构和功能。为此，我们将：AIM 1。通过使用X射线晶体学和分子建模技术，分析四种耐多种药物“签名突变体”（807，3761，1385，769）的抑制剂复合物的结构。将30个模型的蛋白酶变体配体配合物的结构进行系统的比较，以确定抑制剂或底物结合的变化。三分之一的建模结构将通过X射线晶体学验证。目的2。测量一系列四种耐药性HIV-1蛋白酶（807，3761，1385，769）的一系列四种耐药性HIV-1蛋白酶的药物耐药性指标（KI，KM，VMAX）指标，并将功能性结果与来自AIM 1的结构性抗药性相关联。蛋白酶主动位点扩展假设。如果确认主动部位扩展假设，它将提出一种克服耐药性的逻辑方法。解决为什么标准抑制剂不能抑制某些突变的HIV-1蛋白酶并使用这些信息来设计新的有效化合物的结构问题对人类福利至关重要。