PHARMACOLOGY OF THE HIV VIRAL DNA AND RETROVIRAL INTEGRASES

为了进一步扩大发展目标的数量 在抗逆转录病毒药物中，我们正在研究潜在的艾滋病毒整合酶 使用体外整合酶检测的抑制剂。活性化合物为 在NCI抗HIV药物中提交测试其抗病毒活性 屏幕上。相反，我们正在测试屏幕上的活性化合物 用于抑制HIV1整合酶。 药物与HIV1整合酶的分子相互作用如下 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HIV1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 目的：确定HIV-1整合酶的药物结合位点(S)。按顺序 来检验某些抑制物干扰的假设 选择性地与酶的特定功能结构域(锌 Finger，催化区，DNA结合域)，基因改变的HIV1 使用整合酶。HIV整合酶抑制剂的发现可能会提供 抗逆转录病毒治疗的新策略和基础知识 药物与酶的相互作用。 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Um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。 用于共结晶研究。我们还证明了鸟苷 四元系衍生物在纳摩尔浓度下是活性的，并表现出 抗病毒活性。在药物化学家的合作下，我们 继续我们对CAPE衍生品的结构活性研究 并从活性化合物中鉴定出新型的抑制剂 显示在NCI屏幕上。最后，开发了一种新的检测方法来监测 整合酶-DNA结合，可用于进一步研究 酶-DNA相互作用和抑制剂的影响。

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 their antiviral activity in the NCI Anti-HIV Drug Screen. Conversely, we are testing active compounds from the screen for HIV1 integrase inhibition. The molecular interactions of drugs with the HIV1 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 HIV1 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 HIV1 integrase. In order to test the hypothesis that some of the inhibitors interfere selectively with a specific functional domain of the enzyme (zinc finger, catalytic region, DNA binding domain), genetically altered HIV1 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 uM range and water-soluble. Nucleotides are attractive as lead compounds for co-crystallization studies. We have also shown that guanosine quartet derivatives are active at nanomolar concentrations and exhibit antiviral activity. 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. Finally, 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.