Identification of ALPK1 inhibitors

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。

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.