Targeting HIV transcription with RNA-binding small molecules

利用 RNA 结合小分子靶向 HIV 转录

• 批准号: 10453741
• 负责人: Takahiro Yano
• 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 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10453741
• 关键词: Acquired Immunodeficiency Syndrome; Address; Animal Model; Anti-HIV 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; Persons; Pharmaceutical Chemistry; Pharmacodynamics; Phase; Positive Transcriptional Elongation Factor B; Property; Proteins; Proviruses; Quantitative Structure-Activity Relationship; RNA; RNA Binding; RNA Probes; Risk; Side; Simplexvirus; Small Business Innovation Research Grant; Small Molecule Chemical Library; Specificity; 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; 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

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.

概括 当前使用中的抗逆转录病毒疗法（CART）能够抑制HIV-1 无法检测到的水平（<50份/ml），但无法消除潜在的CD4+ T细胞中的病毒。 那就必须无限期地保持卡车，或者如果停产治疗，则可能会风险恢复病毒反弹。 可用的抗HIV药物不能阻止病毒转录或抑制病毒释放 来自蜂窝储层。一类新的抗HIV药物靶向转录可以支撑 目前的推车由于其潜力阻止了潜在感染的CD4+ T细胞中的病毒重新激活， 导致深层延迟状态，随后是这种潜在细胞池的连续衰变 随着时间的推移。可以通过使用破坏者来消除潜在的艾滋病毒水库 HIV的焦油二级结构，这将防止TAT蛋白和其他宿主的结合 转录所需的因素。我们建议鉴定特异性结合的小分子 并破坏艾滋病毒焦油发夹中的顶循环或侧面凸起，导致抑制trans- 病毒启动子和病毒复制的激活。 Vironika LLC开发了新方法 并鉴定与结构化病毒RNA相互作用的小分子的测定。例如， Vironika开发了均质时间分辨荧光（HTRF）的应用 免疫测定），α屏幕（供体/受体珠）和基于热循环的荧光 共振能量传递（FRET），可以实现小型筛选（HTS） 分子以鉴定HSV和EBV潜在感染的抑制剂。包含小说的图书馆和 专有的小分子具有新的医学化学可能性 RNA探针代表焦油发夹。基于细胞的测定将用于研究抗病毒 选定的命中化合物的活性。该提案最终产生的产品是 小分子有选择地结合并破坏艾滋病毒焦油的二级结构 抑制病毒表达所需的TAT和/或P-TEFB的结合。安全的， 靶向HIV TAR/TAT或TAR/P-TEFB相互作用的有效的小分子剂将 不可避免地会改变当前的临床实践，并可能使全球控制该疾病。