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; 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.

该财政年度该项目的目的是建立高通量筛选（HTS）测定法，以评估Alpk1在其配体ADP-Heptose刺激后通过ALPK1评估磷酸化。具体而言，该测定法利用了表达ALPK1的市售HEK293 NF-KB报告细胞系。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，该活性独立于AlPK1激活NF-KB报告基因。此外，将使用表达相同报告基因的ALPK1 KO细胞系来确认途径活性的化合物。除其他参数外，该测定法已针对化合物和配体孵育时间进行优化。