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）和细胞辅因子LEDGF/p75之间的相互作用对有效的病毒复制很重要。我们进一步假设，这种相互作用的抑制剂具有抗病毒活性，诱导抗病毒耐药的可能性低，并且应该具有低毒性。针对必需的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破坏药物将显示与HAART鸡尾酒组分的协同相互作用。由于这些设计的药物靶向不同于活性位点的位点，因此它们也将与目前正在开发的靶向酶活性位点的其他IN抑制剂显示协同作用。长期接受HAART治疗的患者通常会产生耐药病毒株。特异性针对IN-LEDGF/p75相互作用的药物代表了完全不同类别的化合物，并且靶向病毒生命周期中的不同阶段。对逆转录酶、蛋白酶或甚至IN抑制剂表现出抗性的病毒株仍将对这类治疗剂敏感。此外，对潜在的IN-LEDGF/p75破坏药物具有抗性的病毒株的出现速率可能比传统的抗病毒治疗剂慢得多，因为界面包括具有低得多的遗传变异性的细胞蛋白。IN辅因子的研究和潜在的IN-辅因子破坏药物的设计是一个新兴的领域，具有重大发展的潜力。在此之前，我们建立了稳定表达IN的真核293 T细胞系，并将LEDGF/p75鉴定为IN的重要细胞辅因子。最近，我们开发了一种高通量的检测方法来筛选LEDGF/p75-IN相互作用的抑制剂，并鉴定了一系列新的抑制剂。在这个建议中，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。具体而言，我们建议：目标1。设计LEDGF/p75-IN相互作用和Aim 2的新型抑制剂。验证我们的前10种化合物作为IN-LEDGF/p75相互作用的真正抑制剂。这项研究的成功完成将表明，这种创新的策略为抗逆转录病毒治疗和进一步询问LEDG/p75通路提供了新的靶标和抑制剂。通过扩展，这种方法可以用于其他疾病，其中蛋白质-蛋白质相互作用可以被安全地抑制而不影响细胞功能。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。这些相互作用的选择性抑制提供了一种独特的策略，以设计新的和安全的抗病毒活性药物。

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.