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.

报告期间该项目的目的是建立高通量筛选测定法，以评估ALPK1在其配体ADP-Heptose刺激后通过ALPK1评估磷酸化。该测定法利用了表达ALPK1的市售HEK293 NF-KB记者细胞系。在存在或不存在ADP-Heptose的情况下，将评估候选化合物的报告。随后将在TNF-Alpha的存在下评估化合物的目标活性，该目标活性独立于ALPK1激活NF-KB记者。该测定正在针对小分子库进行筛选，并确定了潜在的命中。将开发生化测定以确认目标活动。