Potency assay (off target):
Selectivity of cpd 23 on a proteome-wide scale was evaluated using the isoTOP ABPP approach. Live MDA-MB-231 cells with cpd 23, its companion negative control cpd 28, or DMSO vehicle for 3 h at 1 μM concentration, followed by addition of 100 μM iodoacetamide-alkyne (IAA) as a clickable and promiscuous, cysteine-reactive probe.
Probe Selectivity in
Vitro:
For both cpd 23 and the negative control cpd 28, was found that, overall, less than 1% of the detected proteins were competed with a ratio of R > 4.5. For cpd 23, only two targets (RBM12B_C204, PLIN3_C39, and PLIN3_C60) were above the threshold out of a total of more than 3000 identified sites, suggesting good selectivity. In case of the KEAP1 negative control cpd 28, only RBM12B_C204 was competed with a ratio >4.5.
