DEVELOPMENT OF NEW APPROACHES TO INHIBIT GROWTH OF HIV

开发抑制艾滋病病毒生长的新方法

• 批准号: 2073417
• 负责人: PATRICK O. BROWN
• 金额: $ 19.24万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-03-01 至 1998-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2073417
• 关键词: DNA footprinting; RNA directed DNA polymerase; antiviral agents; drug design /synthesis /production; drug screening /evaluation; enzyme inhibitors; enzyme mechanism; enzyme structure; enzyme substrate; genetic library; human immunodeficiency virus 1; integrase; murine leukemia virus; site directed mutagenesis; tissue /cell culture; transposon /insertion element; virus DNA; virus infection mechanism; virus protein; virus replication; yeasts

The early intracellular events in human immunodeficiency virus (HIV) infection, leading to the establishment of the integrated provirus, remain inadequately understood. We hypothesize that many steps in this process, presently unrecognized or unexplored, could provide new targets against which antiviral drugs might be developed. One of the better understood steps in early infection is integration. Integration has clearly been shown to be required for HIV replication, and the biochemistry of integration is understood in broad outline. Yet we are still far from a detailed understanding of integration, and consequently the current approach to developing drugs targeted at integrase may be naive. Indeed, consideration of the place integration occupies in the sequence of events leading to integration in vivo suggests that the most straightforward approach to blocking integration, based on inhibitors of the catalytic activities of integrase, may not be the most effective strategy for preventing its occurrence in an actual infection. The goal of the proposed work is simply to explore and develop the most promising avenues toward discovery of new antiviral agents directed in integration and other early steps in the infection process. Four projects are planned: 1. Continued basic investigations of the biochemistry of integration, aimed at developing a complete description of the regulation, specificity, biochemical mechanism and structure of integrase, and other possible actors in the process. As an integral part of this basic work we will continue to develop assays to test each activity that we can isolate, for possible use in primary or secondary screens of candidate drugs. 2. Exploring new strategies for antiviral agents directed at integrase and testing their feasibility by using mutations in integrase as surrogates for inhibitors. 3. Uncovering new targets for antiviral agents in early infection, using two genetic strategies. We will use a powerful new genetic approach to screen for new mutations in the gag and pol coding regions that impair specific steps in early infection. We will also continue work in progress, using the yeast two-hybrid system to screen for cellular proteins that interact with the Gag or Pol proteins of HIV or murine leukemia virus (MLV). 4. We will define the mechanism of action of integrase inhibitors identified by the ongoing random screens being conducted by the Parke-Davis group.

人类免疫缺陷病毒（HIV）的早期细胞内事件 感染，导致整合的前病毒的建立， 仍然没有被充分理解。 我们假设这其中的许多步骤 目前尚未认识或探索的过程可以提供新的目标 抗病毒药物可能会被开发出来。 一个更好的 早期感染的步骤是整合。 一体化 显然是HIV复制所必需的， 生物化学的整合是广义上理解的。 然而我们 对集成的详细了解还很远，因此 目前开发针对整合酶的药物的方法可能 天真了 事实上，考虑到一体化在 导致体内整合的事件序列表明， 直接的方法来阻止整合，基于抑制剂， 整合酶的催化活性可能不是最有效的 预防其在实际感染中发生的策略。 所提议的工作的目标仅仅是探索和发展最 发现新的抗病毒药物的有希望的途径， 整合和感染过程中的其他早期步骤。 四 计划项目：1. 继续进行基本调查， 生物化学的整合，旨在开发一个完整的描述 的调节，特异性，生化机制和结构， 整合酶，以及该过程中其他可能的参与者。 作为整体部分 在这项基础工作中，我们将继续开发检测方法， 我们可以隔离的活动，可能用于主要或次要 筛选候选药物。 2. 探索抗病毒的新策略 针对整合酶的试剂，并通过使用 整合酶的突变作为抑制剂的替代物。 3. 发现新 抗病毒药物在早期感染中的靶点，使用两种基因 战略布局 我们将使用一种强大的新遗传方法来筛选 gag和pol编码区的新突变， 早期感染的步骤。 我们还将继续进行工作， 酵母双杂交系统来筛选相互作用的细胞蛋白 与HIV或鼠白血病病毒（MLV）的Gag或Pol蛋白结合。 4. 我们将确定整合酶抑制剂的作用机制， 帕克-戴维斯小组正在进行的随机筛选。