Targeting HIV transcription with RNA-binding small molecules

利用 RNA 结合小分子靶向 HIV 转录

• 批准号: 10323077
• 负责人: Nelson Delgado
• 金额: $ 30万
• 依托单位: VIRONIKA, LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-20 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10323077
• 关键词: Acquired Immunodeficiency Syndrome; Address; Animal Model; Anti-HIV Agents; Antiviral Agents; Apical; Area; Binding; Biochemical; Biological Assay; CD4 Positive T Lymphocytes; Cell Culture Techniques; Cells; Chemicals; Development; Disease; Drug Kinetics; Drug Targeting; Epstein-Barr Virus latency; Failure; Fluorescence; Fluorescence Resonance Energy Transfer; Foxes; Funding; Genetic Transcription; Goals; HIV; HIV Infections; HIV-1; HIV-associated neurocognitive disorder; Health; Human; Human Resources; Immunoassay; Individual; Integration Host Factors; Lead; Libraries; Ligands; Malignant Neoplasms; Methods; Modeling; Patients; Pharmaceutical Chemistry; Pharmacodynamics; Phase; Positive Transcriptional Elongation Factor B; Property; Proviruses; Quantitative Structure-Activity Relationship; RNA; RNA Binding; RNA Probes; Risk; Side; Simplexvirus; Small Business Innovation Research Grant; Small Molecule Chemical Library; Specificity; Structural Protein; Structure; Therapeutic; Time; Transactivation; United States National Institutes of Health; Validation; Viral; Viremia; Virus Activation; Virus Replication; antiretroviral therapy; base; biophysical properties; clinical practice; disorder control; efficacy testing; experience; high throughput screening; inhibitor/antagonist; latent HIV reservoir; latent infection; lead optimization; meetings; novel; novel therapeutics; pharmacophore; prevent; promoter; response; screening; small molecule; small molecule inhibitor; small molecule libraries; targeted agent; tat Protein; viral RNA; viral rebound; virology

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.

总结 目前使用的联合抗逆转录病毒疗法（cART）能够抑制HIV-1， 检测不到的水平（<50拷贝/mL），但不能消除潜伏的CD 4 + T细胞中的前病毒。 因此，患者必须无限期地接受cART治疗，否则如果停止治疗，就有病毒反弹的风险。 现有的抗HIV药物不能阻止前病毒的转录，也不能抑制病毒的释放 从细胞储库中。一种靶向转录的新型抗艾滋病毒药物可以支持 目前的cART由于其在潜伏感染的CD 4 + T细胞中阻断病毒再活化的潜力， 导致深度潜伏状态，随后是这种潜伏细胞池的持续衰减 随着时间消除潜伏的艾滋病毒库可以通过使用干扰剂， HIV的TAR二级结构，这将阻止达特蛋白和其他宿主的结合 转录所需的因子。我们建议鉴定出能特异性结合 并破坏HIV TAR发夹中的顶环或侧凸，从而抑制反式- 病毒启动子的激活和病毒复制。Vironika LLC开发了新的方法 以及鉴定与结构化病毒RNA相互作用的小分子的测定。比如说， Vironika开发了均匀时间分辨荧光（HTRF）的应用 免疫分析）、Alpha筛选（供体/受体珠）和基于热循环仪的荧光分析 共振能量转移（FRET），其使得能够高通量筛选（HTS）小的 分子来鉴定HSV和EBV潜伏感染的抑制剂。图书馆包含小说和 具有新药物化学潜力的专有小分子将使用 代表TAR发夹的RNA探针。基于细胞的检测将用于研究抗病毒药物 选定的命中化合物的活性。该提案最终产生的产品是 选择性结合并破坏HIV TAR二级结构的小分子， 抑制病毒表达所需达特和/或P-TEFb的结合。安全， 靶向HIV TAR/达特或TAR/P-TEFb相互作用的有效的小分子试剂将 不可避免地改变了目前临床实践，并可能使这种疾病得到全球控制。