Biochemical Mechanism of HIV DNA Integration

项目总结/摘要 艾滋病毒/艾滋病仍然是全球范围内的一种使人衰弱的疾病，近年来在美国的年轻人中感染 增加。尽管广泛的努力致力于HIV疫苗和治疗研究，但这些方法 还没有产生用于常规临床使用的候选物。相比之下，联合抗逆转录病毒疗法（cART）已被 自20世纪90年代中期引入临床以来，用于减少疾病负担和死亡率。 推荐的cART制剂含有整合酶抑制剂，该抑制剂抑制酶活性位点及其代谢。 链转移活性（整合酶链转移抑制剂或TBI）。尽管他们取得了巨大的成功， 对第二代INSTI耐药的发生率正在增加，并且随着 这些药物已在全球推广使用。证实了活性位点和变构位点抑制剂的成功， 逆转录酶，临床将大大受益于第二类整合酶的加入 抑制剂，如变构整合酶抑制剂（ALLINI）。本供资周期的赠款 对理解临床前ALLINI化合物的作用机制做出了开创性贡献， 化合物目前正在制药公司开发中。在这份资助申请中，我们将继续 对ALLINI的作用机制进行分类，这是临床研究之前所需的关键基础信息。 推广和临床耐药性。这项研究将部分集中在行动的机制， 整合酶结合蛋白透镜上皮衍生生长因子（LEDGF）/p75，有助于引导病毒 整合的活性基因。特别是，一些研究得最好的ALLINI化学型是有效的抑制剂 LEDGF/p75-整合酶结合相互作用。受LEDGF/75结合位点ALLINI成功的启发 化合物，我们现在将详细描述其他宿主因子的相互作用， 整合酶正如引起多效性复制灾难的大量突变所证明的那样， 整合酶对变化极其敏感。新型宿主因子-整合酶复合物的表征将 为未来抗逆转录病毒抑制剂的开发确定新的靶点。

PROJECT SUMMARY/ABSTRACT HIV/AIDS remains a debilitating disease globally, with infections among young adults in the US in recent years increasing. Although extensive efforts are dedicated to HIV vaccine and cure research, these approaches have yet to yield candidates for routine clinical use. By contrast, combination antiretroviral therapy (cART) has been used to reduce disease burden and mortality since its introduction into the clinic in the mid-1990s. Recommended cART formulations contain an integrase inhibitor that inhibits the enzyme active site and its strand transfer activity (integrase strand transfer inhibitor or INSTI). Despite their resounding success, incidence of resistance to second-generation INSTIs is increasing, and will predictably increase further as these drugs are rolled out for global usage. Paralleling the success of active site and allosteric site inhibitors of the reverse transcriptase enzyme, the clinic will benefit greatly from the addition of a second class of integrase inhibitor, such as allosteric integrase inhibitors (ALLINIs). This grant over the current funding cycle made seminal contributions to understanding the mechanism of action of pre-clinical ALLINI compounds, and such compounds are today in development at pharmaceutical companies. In this grant application we will continue to categorize the mechanism of ALLINI action, which is critical basic information required in advance of clinical rollout and clinical drug resistance. This research will in part be focused on the mechanism of action of the integrase binding protein lens epithelium-derived growth factor (LEDGF)/p75, which helps to guide the virus to active genes for integration. In particular, some of the best-studied ALLINI chemotypes are effective inhibitors of the LEDGF/p75-integrase binding interaction. Inspired by the success of LEDGF/75 binding site ALLINI compounds, we will now characterize in detail interactions of additional host factors that are shown to bind integrase. As evidenced by the large variety of mutations that cause pleiotropic replication catastrophe, HIV-1 integrase is extremely sensitive to change. Characterization of novel host factor-integrase complexes will define new targets for future antiretroviral inhibitor development.