PROTEASE INHIBITOR DESIGN AGAINST MDR HIV VARIANTS

针对耐多药艾滋病毒变异体的蛋白酶抑制剂设计

• 批准号: 6947599
• 负责人: LADISLAU Christopher KOVARI
• 金额: $ 22万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-15 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6947599
• 关键词: AIDS therapy; HIV infections; X ray crystallography; antiAIDS agent; antiviral agents; calorimetry; chemical binding; drug design /synthesis /production; drug screening /evaluation; enzyme structure; gene mutation; human immunodeficiency virus 1; multidrug resistance; protease inhibitor; protein structure function; surface plasmon resonance

DESCRIPTION (provided by applicant): The development of HIV virus strains that are resistant to existing protease inhibitors is one of the major obstacles to successful long-term antiretroviral therapy. We are investigating the structural changes of HIV-1 proteases from naturally occurring clinical variants that have become resistant to licensed protease inhibitors. The long-term goal of this project is to study the structural basis of HIV protease drug resistance and to use that knowledge to devise strategies to overcome resistance. The hypothesis being tested is that HIV-1 protease multidrug resistance is associated with mutations that expand the active site cavity 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 the project is to perform structure-function studies with a set of four multidrug-resistant HIV-1 protease clinical isolates. To accomplish this we will: Aim 1. Define the structural basis for multi-drug resistance in MDR HIV-1 protease variants and perform functional analyses of MDR HIV-1 protease variants to develop three dimensional structure-activity relationships that explain, in terms of protein-ligand interactions, why the drugs inhibit these structures less well compared to the wild-type. Aim 2. Design, synthesize and test small molecule inhibitors against the MDR HIV-1 protease variants. The HIV-1 protease variants described in Aim 1 will be subjected to X-ray crystallographic and functional studies. Structural studies are focused at discovering the three-dimensional changes in the protease variants. Mutant protease-ligand structures will be compared to wild-type protease-ligand structures. Functional studies of the ligands to the altered protease variants will be performed using surface plasmon resonance (BIACORE), isothermal titration calorimetry (ITC), enzyme assays and virologic drug susceptibility assays. Small molecule ligands that represent lead compounds against the MDR HIV-1 protease will be identified, synthesized and tested for efficacy. Peptide derivatives and peptidomimetic ligands will be constructed in four categories: modified peptides, retro-inverso peptides, peptoids and amide isosteres. Solving the structural problem of why certain mutant HIV-1 proteases are not inhibited by the standard inhibitors and using this information to design new and effective compounds are of a great importance to human welfare.

描述（由申请人提供）： 对现有蛋白酶抑制剂具有抗性的HIV病毒菌株的发展是成功长期抗逆转录病毒疗法的主要障碍之一。我们正在研究自然存在的临床变异体的HIV-1蛋白酶的结构变化，这些变体对许可的蛋白酶抑制剂具有抗性。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。测试的假设是HIV-1蛋白酶多药耐药性与扩大蛋白酶活性位点腔的突变有关，因此抑制剂与酶变体形式的亲和力较低。 该项目的短期目标是使用一组四个耐多药HIV-1蛋白酶临床分离株进行结构功能研究。为此，我们将： 目的1。定义MDR HIV-1蛋白酶变体中多药物抗性的结构基础，并对MDR HIV-1蛋白酶变体进行功能分析，以发展三维结构 - 活性关系，这些关系可以用蛋白质 - 配体相互作用来解释，为什么与野生型相比，这些药物抑制这些结构的抑制作用较少。 AIM 2。设计，合成和测试针对MDR HIV-1蛋白酶变体的小分子抑制剂。 AIM 1中描述的HIV-1蛋白酶变体将经过X射线晶体学和功能研究。结构研究的重点是发现蛋白酶变体的三维变化。突变蛋白酶 - 配体结构将与野生型蛋白酶 - 配体结构进行比较。将使用表面等离子体共振（BIACORE），等温滴定量热法（ITC），酶测定和病毒药物敏感性测定法对改变的蛋白酶变体的配体的功能研究。将鉴定，合成并测试代表针对MDR HIV-1蛋白酶的铅化合物的小分子配体。肽衍生物和肽型配体将分为四类：改性肽，恢复互入的肽，肽和酰胺等等值剂。解决了为什么标准抑制剂不会抑制某些突变的HIV-1蛋白酶并使用这些信息设计新的有效化合物的结构问题，这对人类福利非常重要。