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试验，以评估ALPK 1在其配体ADP-庚糖刺激下的磷酸化。该试验使用表达ALPK 1的市售HEK 293 NF-kB报告细胞系。将在存在或不存在ADP-庚糖的情况下评价候选化合物的报告子活化。随后将在TNF-α存在下评价化合物的靶向活性，TNF-α独立于ALPK 1激活NF-κ B报告基因。正在针对小分子文库筛选该测定法，并已鉴定出潜在的命中。将开发一项生化试验，以确认靶标活性。

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.