Inhibition of HIV-1 Integrase-LEDGF/P75 Interactions

描述(由申请人提供)：我们的中心假设是HIV-1整合酶(IN)和细胞辅助因子LEDGF/p75之间的相互作用对于有效的病毒复制是重要的。我们进一步假设，这种相互作用的抑制剂具有抗病毒活性，诱导抗病毒耐药性的机会很低，并且应该具有低毒性。针对重要的IN-辅因子热点设计的药物将对当前的治疗方案产生重大影响。IN-LEDGF/p75干扰药物将与HAART鸡尾酒的成分显示出协同作用。由于这些设计的药物靶点不同于活性部位，它们也将显示出与目前正在开发的其他IN抑制剂的协同作用，这些抑制剂针对的是酶的活性部位。长期接受HAART治疗的患者往往会出现抗药性病毒株。针对IN-LEDGF/p75相互作用的药物代表了一类完全不同的化合物，并针对病毒生命周期的不同阶段。对逆转录酶、蛋白酶甚至IN抑制剂表现出抗药性的病毒株仍然对这种疗法敏感。此外，对潜在的IN-LEDGF/p75干扰药物产生抗药性的病毒株的出现速度可能会比传统抗病毒药物的出现速度慢得多，因为界面包括一种遗传变异性低得多的细胞蛋白。IN辅因子的研究和潜在的IN辅因子干扰药物的设计是一个具有重大发展潜力的新兴领域。此前，我们建立了稳定表达IN的真核293T细胞系，并确定LEDGF/p75是IN的一个重要的细胞辅助因子。最近，我们发展了一种高通量的方法来筛选LEDGF/p75-IN相互作用的抑制剂，并发现了一系列新的抑制剂。在这项建议中，我们的目标是表征这些抑制剂，特别是它们与IN的相互作用，抗病毒活性，并设计优化的抑制剂，选择性地阻断它们与IN的相互作用。更具体地说，我们建议：目的1.设计新型的LEDGF/p75-IN相互作用抑制剂和目标2.验证我们的前10个化合物是真正的IN-LEDGF/p75相互作用抑制剂。这项研究的成功完成将表明，这一创新策略为抗逆转录病毒治疗和进一步询问ledg/p75途径提供了新的靶点和抑制物。此外，这种方法也可以用于其他疾病，在这些疾病中，蛋白质之间的相互作用可以被安全地抑制，而不会影响细胞功能。公共卫生相关性：HIV-1整合酶与一系列细胞辅助因子相互作用，其中许多相互作用，如与LEDGF/p75的相互作用，对于有效的病毒复制是重要的。选择性地抑制这些相互作用为设计具有抗病毒活性的新型安全药物提供了一种独特的策略。

DESCRIPTION (provided by applicant): Our central hypothesis is that the interaction between HIV-1 integrase (IN) and the cellular co-factor LEDGF/p75 is important for efficient viral replication. We further hypothesize that inhibitors of this interaction have antiviral activity, a low chance of inducing antiviral resistance, and should have low toxicity. Designed drugs targeting an essential IN-cofactor hotspot will have a major impact on current treatment regimens. The IN- LEDGF/p75 disrupting drugs would display synergistic interactions with components of HAART cocktails. Because these designed drugs target spots different from the active site, they would also display synergy with other IN inhibitors presently in development, which target the active site of the enzyme. Patients receiving HAART therapy over long periods often develop drug resistant viral strains. Drugs specific for IN-LEDGF/p75 interactions represent a completely different class of compounds, and target different stages in the viral life cycle. Viral strains exhibiting resistance to reverse transcriptase, protease, or even IN inhibitors would still be susceptible to this class of therapeutics. Additionally, the emergence rate of viral strains resistant to potential IN-LEDGF/p75 disrupting drugs would presumably be considerably slower than that to traditional antiviral therapeutics since the interface includes a cellular protein with much lower genetic variability. The study of IN cofactors and the design of potential IN-cofactor disrupting drugs is an emerging field with the potential for major developments. Previously, we created eukaryotic 293T cell lines stably expressing IN and identified LEDGF/p75 as an important cellular co-factor of IN. Recently, we have developed a high throughput assay to screen for inhibitors of the LEDGF/p75-IN interaction and have identified a series of novel inhibitors. In this proposal our goal is to characterize these inhibitors, especially their interaction with IN, antiviral activity, and design optimized inhibitors selectively blocking their interactions with IN. More specifically we propose: Aim 1. To design novel inhibitors of the LEDGF/p75-IN interaction and Aim 2. To validate our top 10 compounds as bona fide inhibitors of IN-LEDGF/p75 interaction. Successful completion of this study will show that this innovative strategy provides a new target and inhibitor for antiretroviral therapy and for further interrogating the LEDG/p75 pathway. By extension this approach can be exploited in other diseases where protein-protein interactions can be safely inhibited without affecting cellular function. PUBLIC HEALTH RELEVANCE: HIV-1 integrase interacts with a series of cellular co-factors and many of these interactions such as with LEDGF/p75 are important for efficient viral replication. Selective inhibition of these interactions provides a unique strategy to design novel and safe drugs with antiviral activity.