Identification of ALPK1 inhibitors

ALPK1 抑制剂的鉴定

• 批准号: 10919704
• 负责人: Alexey Zakharov
• 金额: $ 10万
• 依托单位: NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10919704
• 关键词: Biochemical; Biological Assay; Blindness; Cell Line; Disease; Fever; Goals; Gout; Headache; Helicobacter pylori; Immune; Inflammation; Inflammatory Bowel Diseases; Inherited; Inhibition of NF-KB activation; Lead; Ligands; Link; Malignant Neoplasms; Mutation; NF-kappa B; Periodicals; Phosphorylation; Phosphotransferases; Reporter; Reporting; Role; Signal Transduction; TNF gene; autoinflammatory diseases; high throughput screening; inhibitor; reduce symptoms; sensor; small molecule; small molecule libraries

The goal of the project during the reporting period has been to establish a high-throughput screening assay to assess phosphorylation by ALPK1 upon stimulation by its ligand ADP-heptose. The assay utilizes a commercially available HEK293 NF-kB reporter cell line expressing ALPK1. Candidate compounds will be evaluated for reporter activation in the presence or absence of ADP-heptose. Compounds will be subsequently evaluated for on target activity in the presence of TNF-alpha, which activates the NF-kB reporter independently of ALPK1. The assay is being screened against small molecule libraries and potential hits have been identified. A biochemical assay will be developed to confirm on target activity.

在报告期内，该项目的目标是建立一种高通量筛选方法，以评估ALPK1在其配体adp -庚糖刺激下的磷酸化作用。该试验使用市售的HEK293 NF-kB报告细胞系表达ALPK1。候选化合物将在adp -庚糖存在或不存在的情况下评估报告基因激活。随后将评估化合物在tnf - α存在下的靶活性，tnf - α独立于ALPK1激活NF-kB报告基因。该检测方法正在针对小分子文库进行筛选，并已确定潜在的命中点。将开发生化试验来确认目标活性。