Fig. 2: Preliminary cPLA2 inhibition screen of 1st Generation virtual screening hits. | npj Drug Discovery

Fig. 2: Preliminary cPLA2 inhibition screen of 1st Generation virtual screening hits.

From: Development of potent, selective cPLA2 inhibitors for targeting neuroinflammation in Alzheimer’s disease and other neurodegenerative disorders

Fig. 2

A A fluorescence-based assay was used to determine the activity of recombinant human cPLA2 incubated with 10 μM test compound relative to a control (substrate, enzyme, no compound) in our initial screen. ASB14780 was also included as an inhibitor positive control. Compounds BRI-50006, BRI-50049, BRI-50054, BRI-50077, and BRI-50125 were among the hits found through this initial screen. B The compounds (10 µM) were mixed with stimuli (TIA: TNFa+rIFN+A23187) and added to astrocytes for 30 minutes. The cell lysate was harvested and used to test the protein levels of phosphorylated and total cPLA2 by Western blot. ASB14780 was used as a positive cPLA2 inhibitor control. C BV-2 microglia cells were pre-treated with different concentrations of BRI-50054, BRI-50077, and BRI-50125 for 30 minutes, then treated with LPS (10 ng/mL) for 16 hours. PGE2 levels in the medium were measured using a PGE2 ELISA assay kit.

Back to article page