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在宿主体内复制和繁殖的三种必需酶）的药物是可用的。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。这三个靶点中的一个可能被证明具有进一步的“可药物”域。尽管体外筛选抗HIV整合酶（INT）的抑制药物一直很困难，因为它的低水溶性和倾向于蛋白质聚集，我们已经发现了一种抗INT的抑制肽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。这种抑制肽与酵母双杂交技术的结合，可能会为发现新的INT抑制化合物提供一个筛选平台。提出的研究目标是获得一个小分子，模仿的机制作用的肽抑制剂的HIV INT。简而言之，具体目的是1)优化靶肽置换法；2)在高通量条件下筛选酵母中破坏肽- hiv INT相互作用的化合物的化学文库；3)筛选最佳候选人进行筛选。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整合酶是这种破坏性病毒复制周期中的一种必需酶。分离出的具有理想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.