Identification of ALPK1 inhibitors

ALPK1 抑制剂的鉴定

• 批准号: 10691144
• 负责人: Alexey Zakharov
• 金额: $ 9万
• 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/10691144
• 关键词: Biological Assay; Blindness; Cell Line; Disease; Fever; Goals; Gout; Headache; Helicobacter pylori; Heptoses; Immune; Inflammation; Inflammatory Bowel Diseases; Inherited; Lead; Ligands; Link; Malignant Neoplasms; Mutation; NF-kappa B; Pathway interactions; Periodicity; Phosphorylation; Phosphotransferases; Reporter; Role; Signal Transduction; TNF gene; Time; autoinflammatory; high throughput screening; inhibitor; reduce symptoms; sensor; small molecule

The goal of the project during this fiscal year has been to establish a high-throughput screening (HTS) assay to assess phosphorylation by ALPK1 upon stimulation by its ligand ADP-heptose. Specifically, 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. Furthermore, ALPK1 KO cell lines expressing the same reporter, will be utilized to confirm compound on pathway activity. The assay is being optimized for compound and ligand incubation time, among other parameters.

本财年该项目的目标是建立一种高通量筛选(HTS)试验，以评估ALPK1在其配体ADP-庚糖刺激下的磷酸化。具体地说，该检测利用了商业上可获得的表达ALPK1的HEK293 NF-kB报告细胞系。候选化合物将在存在或不存在ADP-庚糖的情况下进行报告活化评估。随后将在肿瘤坏死因子-α存在的情况下评估化合物的“靶向”活性，肿瘤坏死因子-α独立于ALPK1激活核因子-kB报告。此外，表达相同报告的ALPK1 KO细胞株将被用来证实化合物在通路上的活性。在其他参数中，该分析正在对化合物和配体孵育时间进行优化。