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产生耐药性的 HIV 病毒株的发展是长期抗逆转录病毒治疗取得成功的主要障碍之一。我们正在研究自然发生的临床变体中 HIV-1 蛋白酶的结构变化，这些变体对许可的蛋白酶抑制剂产生了抗药性。该项目的长期目标是研究艾滋病毒蛋白酶耐药性的结构基础，并利用这些知识制定克服耐药性的策略。正在测试的假设是，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，因此抑制剂与酶的变体形式的结合亲和力较低。 该项目的短期目标是对四种多重耐药 HIV-1 蛋白酶临床分离株进行结构功能研究。为了实现这一目标，我们将： 目标 1. 定义 MDR HIV-1 蛋白酶变体中多药耐药性的结构基础，并对 MDR HIV-1 蛋白酶变体进行功能分析，以建立三维结构-活性关系，从蛋白质-配体相互作用的角度解释为什么与野生型相比，药物对这些结构的抑制效果较差。 目标 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。 目标 1 中描述的 HIV-1 蛋白酶变体将接受 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。将突变型蛋白酶-配体结构与野生型蛋白酶-配体结构进行比较。将使用表面等离子共振 (BIACORE)、等温滴定量热法 (ITC)、酶测定和病毒学药物敏感性测定对改变的蛋白酶变体的配体进行功能研究。将鉴定、合成代表针对 MDR HIV-1 蛋白酶的先导化合物的小分子配体并测试其功效。肽衍生物和拟肽配体将分为四类：修饰肽、逆反肽、类肽和酰胺电子等排体。解决为什么某些突变型 HIV-1 蛋白酶不受标准抑制剂抑制的结构问题，并利用这些信息设计新的有效化合物，对人类福祉非常重要。