Development and characterization of HIV-1 Tat degraders

HIV-1 Tat 降解剂的开发和表征

基本信息

批准号：
10483950
负责人：
Susana T Valente
金额：
$ 30万
依托单位：
THIMBLE THERAPEUTICS, INC.
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10483950
关键词：
Affinity Animal Model Anti-HIV Agents Anti-Retroviral Agents Apical Automobile Driving Binding Sites Biochemical Biological Assay CD4 Positive T Lymphocytes Calorimetry Cell Survival Cell model Cells Chemicals Chimeric Proteins Clinical Clinical Trials Collaborations Complement Complex Confocal Microscopy DNA Development Disease Progression Docking Electrophoretic Mobility Shift Assay Epigenetic Process Feedback Florida Future Gene Silencing Genetic Transcription Genome Goals HIV HIV tat Protein HIV-1 Hela Cells Human In Vitro Individual Interruption Laboratories Lead Libraries Luciferases Mediating Metabolic Modification Molecular Molecular Computations Names Nuclear Parents Pathway interactions Persons Pharmaceutical Preparations Pharmacology Production Property Proviruses RNA Research Research Institute Role Shock Specificity Structure Structure-Activity Relationship System TNF gene Techniques Testing Tetanus Helper Peptide Therapeutic Therapeutic Index Therapeutic Intervention Tissues Titrations Toxic effect Transactivation Transcript Validation Viral Viral reservoir Virus Virus Activation Virus Replication analog antiretroviral therapy base blood-brain barrier disruption chemical property chromatin immunoprecipitation clinical candidate commercialization cortistatin cost cost effective cytotoxicity detection limit disease transmission epigenetic silencing follow-up high throughput screening high-throughput drug screening humanized mouse improved inhibitor interest molecular dynamics mouse model mutant neuroinflammation neurotoxicity novel novel drug class novel strategies particle pre-clinical prevent promoter small molecule targeted treatment tat Protein viral rebound viral resistance

项目摘要

Abstract Despite effective antiretroviral therapy (ART), latent proviruses can reinitiate viral production upon cell stimulation or treatment interruption. The viral Tat protein enhances transcript elongation from the HIV-1 promoter, controlling the switch between latency and active viral production. The block-and-lock functional cure aims at the transcriptional silencing of the viral reservoir, rendering suppressed HIV promoters extremely difficult to reactivate from latency. The Tat inhibitor, didehydro-cortistatin A (dCA) was used to prove this concept. Combining dCA with ART inhibits transcription and blocks viral rebound upon treatment interruption, as the promoter becomes epigenetically repressed. dCA defines a novel class of drugs that can silence and maintain a transcriptionally inactive HIV promoter, offering a novel approach in the treatment of HIV. Tat is very attractive target for therapeutic intervention because: 1) is expressed early during virus replication; 2) no cellular homologs; 3) Tat inhibitors block the feedback loop necessary for viral amplification; 4) epigenetic modifications accumulate at the HIV promoter rendering reactivation less likely. Tat is also known for its role in neurotoxicity, blood-brain barrier disruption, and neuroinflammation. Thus, the immense interest in the development of Tat inhibitors to complement ART. The major hurdle towards advancing dCA into clinical trials is the cost of producing large quantities of this molecule due to its complex structure. Additional clinical candidates, structurally distinct from dCA, that embody equivalent bioactivity are needed in the pre-clinical pipeline. We optimized a cell-based Tat transactivation assay to use in high throughput screening (HTS), with dCA as control. We combined appropriate counter-screens and a wealth of techniques to quickly discard small molecules that are not Tat specific. The HTS of 369,203 compounds was completed by Southern Research (SR), yielding three compounds, TT-44951, TT-44881 and TT-44863, with therapeutic index (TI) varying from 51.2 to 181.1 and good chemical properties. In this application, we propose to perform hit validation and characterization of chemically synthesized analogs generated by Thimble Therapeutics, during the compound progression pathway (CPP), to expand our panel of Tat inhibitors to commercialize this novel class of compounds. We propose the following aims: Specific Aim 1. Validate Tat inhibitors based on disruption of Tat HIV-1 LTR transactivation. Specific Aim 2. Characterize the mechanism of action of selected hits. At the end of this study we expect to: (a) have identified small molecules that will specifically inhibit Tat in cell-based assays; (b) have adequate metabolic stability and PK properties for future pharmacological assessment in animal models and eventually in human clinical trials.

抽象的尽管有史刺激或治疗中断。病毒TAT蛋白增强了HIV-1的转录伸长启动子，控制潜伏期和活动病毒产生之间的切换。阻止锁定功能治疗目的是针对病毒储层的转录沉默，使抑制的艾滋病毒启动子极为困难从延迟重新激活。 TAT抑制剂Didehydro-Cortistatin A（DCA）用于证明这一概念。将DCA与ART结合抑制转录并在治疗中断时阻止病毒反弹，因为发起人在表观遗传上受到压抑。 DCA定义了一种可以使和维持的新型药物具有转录的无活性HIV启动子，为艾滋病毒治疗提供了一种新颖的方法。 TAT是治疗干预的非常有吸引力的靶标，因为：1）在病毒复制期间早期表达； 2）无细胞同源物； 3）TAT抑制剂阻止病毒扩增所需的反馈回路； 4）表观遗传学在HIV启动子渲染的重新激活下积累的修饰可能较小。塔特还以其在神经毒性，血脑屏障障碍和神经炎症。因此，对开发TAT抑制剂以补充艺术。将DCA推进临床试验的主要障碍是生产大量的成本分子由于其复杂的结构。在结构上与DCA不同的其他临床候选物体现在临床前管道中需要等效生物活性。我们优化了基于细胞的TAT反式激活测定法，以在高吞吐量筛选（HTS）中使用DCA作为控制。我们结合了适当的柜台和大量技术来快速丢弃小分子不是特定的。南方研究（SR）完成了369,203种化合物的HTS，产生三种化合物TT-44951，TT-44881和TT-44863，治疗指数（TI）从51.2到181.1不等和良好的化学特性。在此应用程序中，我们建议执行命中验证和表征在化合物进程途径期间由顶针治疗产生的化学合成类似物（CPP），扩展我们的TAT抑制剂小组，以将这种新型化合物类别商业化。我们建议以下目的：具体目标1。基于TAT HIV-1 LTR反式激活的破坏来验证TAT抑制剂。特定目的2。表征选定命中的作用机理。在这项研究结束时，我们期望：（a）已经确定了小分子，这些分子会特别抑制基于细胞的测定中的TAT；（b）具有足够的代谢稳定性和未来的PK特性动物模型和人类临床试验中的药理评估。