Identification of REV-ERB inverse agonists for cancer immunotherapy

用于癌症免疫治疗的 REV-ERB 反向激动剂的鉴定

基本信息

批准号：
9923624
负责人：
Louis Daniel Scampavia
金额：
$ 64.3万
依托单位：
SCRIPPS FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-01 至 2023-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9923624
关键词：
Agonist Automation Binding Binding Proteins Bioinformatics Biological Assay Biological Response Modifiers Cells Collection Consensus Sequence Crystallography DNA Data Development Disease Equilibrium Exhibits Fluorescence Resonance Energy Transfer Frequencies Genes Genetic Genetic Transcription Goals Immune checkpoint inhibitor Immunologic Surveillance Immunotherapy In Vitro Inflammatory Inflammatory Bowel Diseases Interleukin-17 Lead Libraries Ligand Binding Ligands Malignant Neoplasms Mediating Multiple Sclerosis Nuclear Receptors Pharmaceutical Chemistry Pharmacology Phase I/II Clinical Trial Plasma Proteins Play Positioning Attribute Property Proteins Regimen Regulation Reproducibility Research Resistance Response Elements Role Running Series Solubility Specificity Structure Structure-Activity Relationship T-Lymphocyte Testing Therapeutic Tissues Toxic effect Transcription Coactivator Transcription Repressor Tumor Immunity Validation anti-PD-1/PD-L1 base cancer immunotherapy chemical synthesis cheminformatics counterscreen cytokine cytotoxicity design drug discovery drug metabolism fight against follow-up genetic approach high throughput screening in vivo lead optimization meetings member miniaturize mouse model neoplasm immunotherapy novel novel strategies overexpression patient subsets pharmacophore receptor response scaffold screening small molecule therapeutic target tool transcription factor tumor

项目摘要

PROJECT SUMMARY While immunotherapy is a breakthrough in our fight against cancer, only a subset of patients display robust and long-lasting responses highlighting the critical need for novel approaches to be used alone or in combination with current therapeutic regimens. TH17 cells, which express the lineage defining transcription factor RORgt, have emerged as targets to enhance antitumor immunity, with RORg agonists currently in Phase 1/2 clinical trials. The transcriptional repressors REV-ERBa and REV-ERBb are often co-expressed in the same tissues as RORg(t) and bind the same DNA response elements, which suggests mutual cross talk and co-regulation of their target genes. Our preliminary data demonstrates that the REV-ERBs are ligand-regulated transcription factors and play critical roles in TH17 responses, including regulation of IL-17A expression through competition with RORgt at their shared DNA consensus sequence(s), and regulation of RORgt expression itself. We have generated small molecule ligands that enhance the REV-ERBs repressive function and inhibit TH17 cell development in vitro and in vivo. Thus, we hypothesize that identification of ligands that inhibit the REV-ERBs repressive activity will enhance TH17 responses and antitumor immunity. In order to identify ligands that inhibit REV-ERB activity, we have designed a high-throughput screening (HTS) compatible primary assay that detects direct ligand binding to each receptor. Our goal is to implement a full HTS-campaign using the Scripps Institutional Drug Discovery Library (SDDL) to identify, validate, and characterize potent small molecule inverse agonists of REV-ERB activity. To achieve our goal, we will miniaturize our assay into a 1,536-well plate format and upon meeting HTS automation criteria, carry out a “full-deck” HTS campaign to screen the >640,000 compounds in the SDDL (Aim 1). Cheminformatic analysis of “hits” will help identify the most promising leads by structural clustering, bioinformatics analysis of compounds to determine promiscuity, and scaffold analysis to determine ease of chemical synthesis and tractability for further medicinal chemistry efforts to perform structure- activity relationship studies. In Aim 2, we will use a cascade of follow up assays to validate screening hits, determine specificity, and begin to understand mechanism of action of REV-ERB-mediated transcriptional activity. Finally, in Aim 3, validated HTS hits will be advanced to early medicinal chemistry for lead optimization and characterization of novel negative regulators of REV-ERBa/b. We expect that completion of this application will deliver multiple structurally distinct REV-ERB inverse agonists that exhibit suitable levels of cellular activity, potency, and selectivity. Our collaborative research team has a strong track record of performing high-throughput screens, selective optimization of scaffolds, and in vitro and in vivo characterization of compounds. Collectively, our screening approach puts us in a unique position to identify, validate, and characterize novel REV-ERB- selective small molecule inverse agonists for the study of the REV-ERB’s function in enhancing TH17 responses and to determine whether targeting these receptors is a viable option for immunotherapy.

项目摘要虽然免疫疗法是我们与癌症斗争的突破，但只有一部分患者表现出强大以及持久的反应突出了对单独使用或组合使用新颖方法的关键需求具有当前的治疗方案。表达谱系定义转录因子rorgt的Th17细胞，已成为增强抗肿瘤免疫学的靶标，目前有Rorg脚踝处于1/2阶段试验。转录表示Rev-Erba和Rev-erbB通常在同一组织中共表达如rorg（t）并结合相同的DNA响应元素，这表明相互交叉谈话和共同调节他们的靶基因。我们的初步数据表明Rev-ERB是配体调节的转录因素并在TH17响应中起关键作用，包括通过竞争调节IL-17A表达在RORGT共享的DNA共有序列（S）和RORGT表达本身的调节中。我们有产生的小分子配体增强了Rev-ERB的反射功能并抑制Th17细胞体外和体内发育。这是我们假设抑制Rev-erbs的配体的识别抑制活性将增强TH17反应和抗肿瘤免疫力。为了识别抑制的配体 Rev-erb活动，我们设计了一个高通量筛选（HTS）兼容的主要测定法直接配体与每个接收器结合。我们的目标是使用Scripps实施完整的HTS竞争机构药物发现库（SDDL）以识别，验证和表征有效的小分子倒数 Rev-ERB活动的激动剂。为了实现我们的目标，我们将使我们的评估小型化成1,536孔板格式并在符合HTS自动化标准后，进行“全甲板” HTS运动，以筛选> 640,000 SDDL中的化合物（AIM 1）。 “命中”的化学信息分析将有助于确定最有希望的潜在客户通过结构聚类，化合物的生物信息学分析以确定滥交，并分析脚手架分析确定化学合成的便利性和易处理性，以进行进一步的药物化学努力，以执行结构 - 活动关系研究。在AIM 2中，我们将使用一系列后续评估来验证筛选命中，确定特异性，并开始了解Rev-ERB介导的转录的作用机理活动。最后，在AIM 3中，经过验证的HTS命中将推进早期的医学化学以进行铅优化 Rev-Erba/b的新型负调节剂的表征和表征。我们希望完成此申请将提供多种结构上不同的Rev-ERB反向激动剂，表现出适当水平的细胞活性，效力和选择性。我们的协作研究团队在执行高通量方面具有良好的记录筛选，脚手架的选择性优化以及化合物的体外和体内表征。共同我们的筛选方法使我们处于独特的位置，以识别，验证和表征新颖的Rev-erb- 选择性小分子反激动剂研究Rev-ERB在增强Th17响应方面的功能并确定靶向这些受体是否是免疫疗法的可行选择。