Technology for isolation of Integrase Inhibitors for Therapeutics against Human I

描述(申请人提供)：目前，全球估计有4,000万人感染艾滋病毒/艾滋病；自1980年S发现以来，已有2,200万人死亡。抑制逆转录酶(RT)、蛋白酶和整合酶--艾滋病毒在宿主中复制和繁殖的三种关键酶--的药物已可用。然而，病毒耐药性和药物副作用要求开发新的艾滋病毒/艾滋病治疗方法。这三个目标中的一个可能会被证明有更多的“可用药”领域。虽然由于HIV整合酶(INT)的低水溶性和蛋白质聚集倾向，在体外筛选抑制HIV整合酶(INT)的药物一直是困难的，但我们使用酵母双杂交试验发现了一种抗INT的抑制肽。与目前的链转移抑制剂相比，该多肽具有独特的作用模式。这种抑制肽与酵母双杂交技术相结合，可能成为一个筛选平台，从而发现新的INT抑制化合物。这项拟议的研究的目标是获得一种小分子，它模仿HIV INT的多肽抑制剂的作用机制。简而言之，具体目标是1)优化靶肽置换分析；2)在高通量条件下筛选破坏酵母中肽-HIV int相互作用的化合物的化学文库；以及3)选择最佳候选者进行筛选。与公共卫生相关：拟议的研究描述了分离抑制HIV整合酶的新药的方法，HIV整合酶是这种毁灭性病毒复制周期中的一种关键酶。分离出的具有良好抑制特性的化合物将被优化用于进一步的临床前研究。

DESCRIPTION (provided by applicant): Currently there are 40 million people estimated to be infected with HIV/AIDS worldwide; 22 million people have died since its discovery in 1980's. Drugs inhibiting the reverse transcriptase (RT), protease, and integrase - three essential enzymes for the replication and propagation of HIV in the host - are available. However, viral resistance and drug side effects necessitate development of new treatments for HIV/AIDS. One target among these three may prove to have further "drugable" domains. Although in vitro screens for inhibitory drugs against HIV integrase (INT) have been difficult due to its low aqueous solubility and propensity toward protein aggregation, we have discovered an inhibitory peptide against INT using a yeast two hybrid assay. The peptide has a unique mode of action than the current strand transfer inhibitors. This inhibitory peptide in conjunction with yeast two hybrid technology may lend itself to a screening platform resulting in the discovery of novel inhibitory compounds of INT. The goal of the proposed research is to obtain a small molecule that mimics the mechanistic effects of a peptide inhibitor of HIV INT. Briefly, the specific aims are 1) to optimize target-peptide displacement assays; 2) to screen chemical libraries under high throughput conditions for compounds that disrupt peptide-HIV INT interaction in yeast; and 3) to select the best candidates for screening. PUBLIC HEALTH RELEVANCE: The proposed research describes methods to isolate novel drugs that inhibit HIV integrase, an essential enzyme in the replication cycle of this devastating virus. Compounds isolated that possess desirable inhibitory properties will be optimized for further pre-clinical studies.