PHARMACOLOGY OF THE HIV VIRAL DNA AND RETROVIRAL INTEGRASES

为了进一步扩大可持续发展的目标 抗逆转录病毒药物，我们正在研究潜在的HIV整合酶 使用体外整合酶检测的抑制剂。活性化合物为 提交NCI抗HIV药物的抗病毒活性测试 屏幕上。相反，我们正在测试屏幕上的活性化合物 HIV-1整合酶抑制。 药物与HIV-1整合酶的分子相互作用 通过使用几种分析方法比较药物效果来进行调查 探索整合反应的不同步骤：1)DNA结合， 2)二核苷酸的裂解和攻击亲核分子的区别使用 (水，3‘-DNA末端，甘油)，3)链转移(整合)， 4)野生型去整合(整合反应的反向) 和截短的HIV-1整合酶。我们的目标是发现新的抗病毒药物 代理商，以确定活性物质的药物筛选是否会导致 到整合酶抑制剂的发现，以评估哪一步 整合反应可被药物(酶齐聚， DNA结合、3‘-加工、DNA链转移、解体)，以及 确定HIV-1整合酶的药物结合位点(S)。按顺序 为了检验某些抑制物选择性干扰的假设 具有酶的特定功能结构域(锌指，催化 区域，DNA结合域)，基因改变的HIV-1整合酶 使用。HIV整合酶抑制剂的发现可能提供新的策略 抗逆转录病毒治疗和药物酶基础知识 互动。 在过去的一年里，我们继续对核苷酸进行研究 抑制剂并鉴定了一系列活性在|M中的二核苷酸 种类繁多，水溶性好。核苷酸作为先导化合物很有吸引力。 用于共结晶和光亲和标记研究。我们还有 表明鸟苷四元衍生物在纳摩尔分子上是活性的。 并显示出抗病毒活性。一种新的化验方法 开发用于监控整合酶与DNA的结合，可用于进一步 研究酶与DNA的相互作用和抑制剂的作用。 在与药物化学家的合作下，我们继续 CAPE衍生物的构效关系研究及鉴定的新化合物 从NCI筛选的活性化合物中筛选出类抑制剂。一个 通过对药效团的搜索，发现了新的酰肼类化合物。 人们正在进行类似的研究。目前的目标是获得具有以下特性的结晶体 最活跃的抑制剂。

In an effort to further extend the number of targets for development of antiretroviral agents, we are investigating potential HIV integrase inhibitors using in vitro integrase assays. The active compounds are submitted for testing antiviral activity in the NCI Anti-HIV Drug Screen. Conversely, we are testing active compounds from the screen for HIV-1 integrase inhibition. The molecular interactions of drugs with HIV-1 integrase are investigated by comparing the drugs effects using several assays exploring different steps of the integration reaction: 1) DNA binding, 2) dinucleotide cleavage and differential use of attacking nucleophiles (water, 3'-DNA terminus, glycerol), 3) strand transfer (integration), 4) dis- integration (reverse of the integration reaction) by wild-type and truncated HIV-1 integrases. Our goals are to discover new antiviral agents, to determine whether drug screening of active agents will lead to the discovery of integrase inhibitors, to evaluate which step of the integration reaction can be inhibited by drugs (enzyme oligomerization, DNA binding, 3'-processing, DNA strand transfer, disintegration), and to determine the drug binding site(s) in the HIV-1 integrase. In order to test the hypothesis that some of the inhibiors interfere selectively with a specific functional domain of the enzyme (zinc finger, catalytic region, DNA binding domain), genetically altered HIV-1 integrases are used. Discovery of HIV integrase inhibitors may provide new strategies for antiretroviral therapy and basic knowledge for drug-enzyme interactions. During the past year, we have continued our studies with nucleotide inhibitors and identified a series of dinucleotides active in the |M range and water-soluble. Nucleotides are attractive as lead compounds for co-crystallization and photoaffinity labeling studies. We have also shown that guanosine quartet derivatives are active at nanomolar concentrations and exhibit antiviral activity. A novel assay has been developed to monitor integrase-DNA binding, that can be used to further investigate the enzyme-DNA interactions and the effects of inhibitors. In collaboration with the medicinal chemists, we have continued our structure- activity studies from CAPE derivatives and identified novel classes of inhibitors from the active compounds of the NCI screen. A novel hydrazide derivative was discovered from pharmacophore searches. Analogs are being made. An immediate goal is to obtain cocrystals with the most active inhibitors.