Identification of ALPK1 inhibitors

National Center For Advancing Translational Sciences

Investigators

Abstract

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.