Technology for isolation of Integrase Inhibitors for Therapeutics against Human I

描述（由申请人提供）：目前全球估计有4000万人感染艾滋病毒/艾滋病;自20世纪80年代发现以来，已有2200万人死亡。抑制逆转录酶（RT）、蛋白酶和整合酶（HIV在宿主中复制和繁殖的三种基本酶）的药物是可用的。然而，病毒的耐药性和药物的副作用需要开发艾滋病毒/艾滋病的新疗法。这三个目标中的一个可能被证明具有进一步的“可用药”领域。虽然在体外筛选针对HIV整合酶（INT）的抑制性药物一直是困难的，由于其低的水溶性和蛋白质聚集的倾向，我们已经发现了一个抑制肽对INT使用酵母双杂交试验。与目前的链转移抑制剂相比，该肽具有独特的作用模式。该抑制肽与酵母双杂交技术相结合可能有助于筛选平台，从而发现新的INT抑制化合物。拟议研究的目标是获得模拟HIV INT肽抑制剂的机制作用的小分子。简而言之，具体目标是：1）优化靶肽置换测定法; 2）在高通量条件下筛选化学文库中破坏酵母中肽-HIV INT相互作用的化合物;和3）选择用于筛选的最佳候选物。公共卫生相关性：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必需酶。将优化分离的具有所需抑制特性的化合物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.