Development of small molecule cGAS inhibitors for repression of dsDNA-triggered interferon expression

开发用于抑制 dsDNA 触发的干扰素表达的小分子 cGAS 抑制剂

基本信息

批准号：
10176388
负责人：
THOMAS TUSCHL
金额：
$ 50.56万
依托单位：
ROCKEFELLER UNIVERSITY
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：
2019-06-10 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10176388
关键词：
Animals Antiviral Agents Autoimmune Autoimmune Diseases Autoimmune Responses Binding Biochemical Biological Assay Biological Process Biology Cancer cell line Cancerous Cell Aging Cell Lineage Cell physiology Cells Cellular Assay Cellular Stress Characteristics Chemicals Chromosomal Instability Collaborations Communities Complex Crystallization Cyclic GMP Cytosol DNA Development Dinucleoside Phosphates Disease Disease model Double-Stranded RNA Drug Industry Drug Screening Enzymes Event Extravasation Genes Genetic Goals Human Immune Industry Standard Inflammatory Innate Immune Response Institutes Institution Intellectual Property Interferon Type I Interferons Invaded Joints Knock-out Laboratories Lead Libraries Mass Spectrum Analysis Mediating Membrane Methods Mitochondria Modeling Mus Myelogenous Myeloid Cells Names Natural Immunity Neoplasm Metastasis Neurodegenerative Disorders Nuclear Nucleic Acids Obesity Parkinson Disease Pathway interactions Patients Periodicity Peripheral Blood Mononuclear Cell Pharmaceutical Chemistry Pharmaceutical Preparations Production Repression Research Personnel Resources Role Rupture STING1 gene Second Messenger Systems Specificity Stimulator of Interferon Genes Structure Systemic Lupus Erythematosus TREX1 gene Testing Therapeutic Toll-like receptors Validation autoinflammatory base biomaterial compatibility cell type cheminformatics cost cost effective cytokine design ds-DNA efficacy testing exodeoxyribonuclease high throughput screening human disease improved inhibitor/antagonist insight interest loss of function mutation luminescence lupus-like macrophage novel novel therapeutics pathogen phosphodiester prevent programs pseudotoxoplasmosis syndrome response scaffold screening sensor small molecule small molecule inhibitor structural biology tool tumorigenesis

项目摘要

Cyclic GMP-AMP synthase (cGAS) is the primary sensor for cytosolic dsDNA, producing the cyclic dinucleotide cGAMP, a second messenger initiating cytokine production in subsets of myeloid-lineage cell types and responsible for providing innate immunity. Aberrant cytosolic dsDNA contributes to inflammatory diseases, suggesting that inhibition of cGAS may be therapeutically beneficial. We recently developed a mass- spectrometry-based high throughput screen (HTS) using mouse cGAS and identified small-molecule inhibitors that yielded the first active and specific inhibitors of cGAS in mouse cellular assays, albeit with no inhibitory activity in human cells. We now developed a faster and more cost effective HTS for identification of human cGAS inhibitors using chemiluminescence and screened 300,000 compounds. Subsequent medicinal chemistry optimization identified potent and specific inhibitors for human cGAS active in major interferon- producing cell types including primary macrophages and PBMCs. We have also solved co-crystal structures of inhibitors with human cGAS, suggesting structure- and computational-guided optimization path for the lead inhibitors. We propose to identify new hit compounds to access additional drug scaffolds through extended drug library screening to continue supporting our medicinal chemistry program. Using the most potent and specific inhibitors, we wish to characterize their inhibitory effect and pathway specificity in a more diverse range of cells. Our long-term goal is to develop therapeutics for cGAS-dysregulation associated diseases. This proposal is organized in 3 aims: (1) Identify new chemical scaffolds by HTS and apply cheminformatic analysis and medicinal chemistry hit optimization. We will use our newly established and validated HTS method for identification of human cGAS inhibitors to screen a newly available library of 100,000 compounds. New hit compounds will be prioritized for further derivatization to optimize potency and drug-like characteristics. (2) Structure/computation-guided design for optimization of new scaffolds and lead human cGAS inhibitors. We will use the insights gained from inhibitor-cGAS co-crystal structures to design, synthesize, and test derivatives to probe for the most potent, and promising drug-like cGAS inhibitor(s). (3) Develop new cellular validation assays for the identification of potent and highly selective cGAS inhibitors. We propose to evaluate and validate new and recently identified inhibitors by assessing their efficacy, selectivity, and biocompatibility using established and new cellular assays including non-myeloid cancer cell lines with chromosomal instability, primary myeloid cells, and patient-derived myeloid cells. Although initially discovered as central dsDNA sensor for antiviral activity, cGAS is also emerging as a player in obesity, neurodegenerative diseases, tumorigenesis, and cancer metastasis. Well-characterized, validated, potent, and specific cGAS inhibitors are needed in the community studying cGAS-STING biology as well as for treating cGAS-related inflammatory diseases!

环状GMP-AMP合酶（CGA）是胞质DsDNA的主要传感器，产生环状二核苷酸 CGAMP，第二iSsenger，在髓样系细胞类型的子集中启动细胞因子的产生和负责提供先天的免疫力。异常的胞质DSDNA导致炎症性疾病，表明对CGA的抑制可能在治疗上是有益的。我们最近开发了一个质量 - 使用小鼠CGA的基于光谱的高吞吐量屏幕（HTS）并鉴定出小分子抑制剂这产生了小鼠细胞测定中CGA的第一个活性和特异性抑制剂，尽管没有抑制人类细胞的活性。现在，我们开发了更快，更具成本效益的HTS来识别人类使用化学发光并筛选300,000种化合物的CGA抑制剂。随后的药用化学优化确定了活跃在主要干扰素中的人CGA的有效抑制剂和特异性抑制剂产生包括原发性巨噬细胞和PBMC在内的细胞类型。我们还解决了共晶结构具有人CGA的抑制剂，暗示了铅的结构和计算引导优化路径抑制剂。我们建议识别新的命中化合物，以通过扩展访问其他药物脚手架药物库筛查以继续支持我们的药物化学计划。使用最有效的特定的抑制剂，我们希望表征它们在更多样化的抑制作用和途径特异性细胞范围。我们的长期目标是开发针对CGAS调节相关疾病的治疗剂。该提案在3个目标中组织：（1）确定HTS的新化学支架并申请化学物质分析和药物化学命中优化。我们将使用我们新建立的经过验证的HTS方法，用于识别人类CGA抑制剂以筛选新的100,000库库化合物。将优先考虑新的命中化合物，以进一步衍生化，以优化效力和类似药物特征。（2）用于优化新支架和铅的结构/计算指导设计人CGA抑制剂。我们将使用从抑制剂-CGAS共晶结构中获得的见解来设计，合成和测试衍生物，以探测最有效和有希望的类似药物的CGA抑制剂。（3）开发新的细胞验证测定法，以识别有效和高度选择性CGA 抑制剂。我们建议通过评估其评估并验证新的和最近确定的抑制剂使用已建立和新的细胞测定法（包括非乳突）的疗效，选择性和生物相容性具有染色体不稳定性，原发性髓样细胞和患者衍生的髓样细胞的癌细胞系。尽管最初被发现是用于抗病毒活性的中央DSDNA传感器，但CGA也在肥胖，神经退行性疾病，肿瘤发生和癌症转移。特殊的，经过验证，在研究CGAS-Sting Biology以及治疗与CGAS相关的炎症性疾病！