Cell experiment:

Cells are seeded at 1,600 cells per well in 384-well white clear-bottom plates. Twenty-four hours after seeding, cells are treated with NMS-859 (eight dilution points, in duplicate) and incubated for an additional 72 h at 37℃ under a 5% CO2 atmosphere. Cells are then lysed, and the ATP content in each well is determined using a thermostable firefly luciferase-based assay as a measure of cell viability. IC50 values are calculated using the percentage of growth of treated cells versus the untreated control[1].

NMS-859 is a potent, covalent VCP (p97) inhibitor, with IC50s of 0.37 and 0.36 μM for wild-type VCP in the presence of 60 μM and 1 mM ATP in cells, respectively.

NMS-859 is a potent VCP inhibitor, with IC50s of 0.37 and 0.36 μM for wild-type VCP in the presence of 60 μM and 1 mM ATP in cells, respectively. NMS-859 shows very weak inhibitory activity against VCPC522T. NMS-859 also suppresses the proliferation of cells, with IC50s of 3.5 μM and 3.0 μM in HCT116 and HeLa cell lines, respectively[1].

[1]. Magnaghi P, et al. Covalent and allosteric inhibitors of the ATPase VCP/p97 induce cancer cell death. Nat Chem Biol. 2013 Sep;9(9):548-56.