Targeting HIV transcription with RNA-binding small molecules

摘要 目前使用的联合抗逆转录病毒疗法(CART)能够抑制HIV-1到 无法检测到的水平(&lt；50拷贝/毫升)，但无法消除潜伏的CD4+T细胞中的前病毒。 因此，患者必须无限期地接受治疗，否则如果停止治疗，病毒可能会反弹。 现有的抗艾滋病毒药物既不能阻止前病毒转录，也不能抑制病毒释放。 来自蜂窝储藏室。一类针对转录的新型抗艾滋病毒药物可能会支持 目前的CART由于有可能阻止潜伏感染的CD4+T细胞中的病毒重新激活， 导致深度潜伏期的状态，随之而来的是这种潜伏池的持续衰退 随着时间的推移。根除潜伏的艾滋病毒蓄积物可以通过使用干扰物 HIV的TAR二级结构，这将阻止Tat蛋白与其他宿主的结合 转录所需的因子。我们建议识别特定结合的小分子 并破坏HIV焦油发夹的顶环或侧凸，导致抑制反式- 病毒启动子的激活和病毒的复制。Vironika LLC已经开发出新的方法 以及鉴定与结构化病毒RNA相互作用的小分子的方法。例如, Vironika开发了均相时间分辨荧光(HTRF)的应用 免疫分析)、α筛选(供体/受体微珠)和基于热循环仪的荧光 共振能量转移(FRET)，使高通量筛选(HTS)成为可能 用于识别单纯疱疹病毒和EB病毒潜伏感染抑制物的分子。包含新奇和 具有新药物化学潜力的专利小分子将使用 代表焦油发夹的RNA探针。基于细胞的分析将被用于研究抗病毒 选定的HIT化合物的活性。这一提议最终产生的产品是 选择性结合并破坏HIV焦油二级结构的小分子，从而 抑制病毒表达所需的TAT和/或P-TEFb的结合。安全， 针对HIV TAR/TAT或TAR/P-TEFb相互作用的有效小分子药物将 不可避免地改变目前的临床实践，并可能实现对这种疾病的全球控制。

Summary Combination antiretroviral therapy (cART) in current use is able to suppress HIV-1 to undetectable levels (<50 copies/mL), but unable to eliminate the provirus in latent CD4+ T cells. Thus, patients must remain under cART indefinitely or risk viral rebound if therapy is discontinued. The available anti-HIV drugs do not prevent transcription from provirus nor inhibit viral release from cellular reservoirs. A new class of anti-HIV drugs targeting transcription could buttress current cART due to its potential to block viral reactivation in latently infected CD4+ T cells, resulting in a state of deep-latency, followed by the continuous decay of this latent pool of cells over time. Eradication of the latent HIV reservoir could be achieved by employing disruptors of HIV’s TAR secondary structure, which would prevent binding of the Tat protein and other host factors required for transcription. We propose to identify small molecules that specifically bind and disrupt the apical loop or side bulge in HIV’s TAR hairpin that result in inhibition of the trans- activation of the viral promoter and virus replication. Vironika LLC has developed new methods and assays to identify small molecules which interact with structured viral RNA. For example, Vironika has developed applications of Homogeneous Time-Resolved Fluorescence (HTRF immunoassay), Alpha Screen (Donor/Acceptor beads) and thermocycler-based Fluorescence Resonance Energy Transfer (FRET), which enable high-throughput screening (HTS) of small molecules to identify inhibitors of HSV and EBV latent infection. Libraries containing novel and proprietary small molecules with potential for new medicinal chemistry will be screened using an RNA probe representing the TAR hairpin. Cell-based assays will be used to investigate the antiviral activity of selected hit compounds. The product that ultimately results from this proposal is a small molecule that selectively binds and disrupts the secondary structure of HIV’s TAR, thereby inhibiting the binding of TAT and/or P-TEFb which are required for viral expression. Safe, efficacious, small molecule agents targeting HIV TAR/Tat or TAR/P-TEFb interaction would inevitably change current clinical practice and possibly enable global control of this disease.