BDP5290 is a potent inhibitor of both ROCK and MRCK, with IC50 values of 5 nM for ROCK1, 50 nM for ROCK2, 10 nM for MRCKα, and 100 nM for MRCKβ.

MRCKβ:100 nM, MRCKα:10 nM , ROCK1:5 nM , ROCK2:50 nM

BDP5290 as a potent MRCK inhibitor.?X-ray crystallography of the MRCKβ kinase domain in complex with BDP5290 revealed how this ligand interacts with the nucleotide binding pocket.?BDP5290 demonstrated marked selectivity for MRCKβ over ROCK1 or ROCK2 for inhibition of myosin II light chain (MLC) phosphorylation in cells.?While BDP5290 was able to block MLC phosphorylation at both cytoplasmic actin stress fibres and peripheral cortical actin bundles, the ROCK selective inhibitor Y27632 primarily reduced MLC phosphorylation on stress fibres.?BDP5290 was also more effective at reducing MDA-MB-231 breast cancer cell invasion through Matrigel than Y27632.

MDA MB 231 or SCC12 cells were plated in a 96 well plate and cultured for 24 hours.?Cells were then cultured for 24 hours in SCC12 medium with DMSO vehicle, 2 μM Y27632 or indicated concentrations of BDP5290 in an IncuCyte ZOOM.?Pictures were taken every 3 hours and confluence was measured using the IncuCyte analysis software.?AlamarBlue was added to the medium and the cells were cultured for an additional day.?Absorbances at 570 nm and at 600 nm were measured to assess cell health.