DEVELOPMENT OF NEW APPROACHES TO INHIBIT GROWTH OF HIV

人类免疫缺陷病毒(HIV)的早期细胞内事件 感染，导致了整合前病毒的建立， 仍然没有得到充分的理解。我们假设这个过程中的许多步骤 目前未被认识到或未被探索的进程可能提供新的目标 抗病毒药物可能会被开发出来。更好的一种 早期感染的理解步骤是整合。集成具有 显然被证明是复制艾滋病毒所必需的，而且 对整合的生物化学有大致的理解。然而，我们是 对整合的详细理解还远远不够，因此 目前开发针对整合酶的药物的方法可能是 天真的。事实上，考虑到整合在 导致体内整合的一系列事件表明，大多数 基于抑制因素的阻止整合的直接方法 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。 拟议工作的目标很简单，就是探索和发展 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 计划开展的项目：1.继续开展基础调查 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 整合酶和其他可能的参与者在这一过程中。作为不可分割的一部分 在这项基本工作中，我们将继续开发化验方法，以检测每一项 我们可以隔离的活动，以便在主要或次要位置使用 筛选候选药物。2.探索抗病毒新策略 针对整合酶的药物，并通过使用 作为抑制剂替代品的整合酶突变。3.发现新的 抗病毒药物在早期感染中的靶点，使用两种基因 战略。我们将使用一种强大的新基因方法来筛选 GAG和POL编码区的新突变会损害特定的 进入早期感染阶段。我们还将继续进行中的工作，使用 酵母双杂交系统筛选相互作用的细胞蛋白 与HIV或小鼠白血病病毒(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查。

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.