Targeting HIV transcription with RNA-binding small molecules

利用 RNA 结合小分子靶向 HIV 转录

基本信息

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
项目类别：
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

项目摘要

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），但无法消除潜伏 CD4+ T 细胞中的原病毒。因此，患者必须无限期地接受 cART，否则如果停止治疗，就会面临病毒反弹的风险。现有的抗 HIV 药物不能阻止原病毒的转录，也不能抑制病毒的释放一种针对转录的新型抗艾滋病毒药物可以支持这种药物。目前的 cART 由于其有可能阻止潜伏感染的 CD4+ T 细胞中的病毒重新激活，导致深度潜伏状态，随后该潜伏细胞池不断衰减随着时间的推移，可以通过使用干扰素来消除潜在的艾滋病毒储存库。 HIV 的 TAR 二级结构，可防止 Tat 蛋白与其他宿主结合我们建议鉴定特异性结合的小分子。并破坏 HIV TAR 发夹的顶端环或侧面凸起，从而抑制反式 Vironika LLC 开发了病毒启动子激活和病毒复制的新方法。以及鉴定与结构化病毒 RNA 相互作用的小分子的测定。 Vironika 开发了均相时间分辨荧光 (HTRF) 的应用免疫分析）、Alpha Screen（供体/受体珠）和基于热循环仪的荧光共振能量转移 (FRET)，可实现小分子的高通量筛选 (HTS) 分子识别 HSV 和 EBV 潜伏感染的抑制剂。将使用以下方法筛选具有新药物化学潜力的小分子代表 TAR 发夹的 RNA 探针将用于研究抗病毒作用。该提案最终产生的产品是选定的热门化合物的活性。选择性结合并破坏 HIV TAR 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。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当前的临床实践，并可能实现对这种疾病的全球控制。