Off target protein and potency: BRD9: 1,000 nM
Potency assay (off target):
NI-57 was screened for binding by DSF against a panel of 47 Bromodomains, and it showed excellent selectivity. All activity was confined to the class IV family of BRDs with ∆Tm < 1 oC stabilization for all non-class IV BRDs except TRIM24 (2.0), CREBBP (1.1) and EP300 (1.1). NI-57 was also tested for Bromodomain selectivity in the BROMOscan panel of 46 binding assays, which showed all potent activity was confined to the class IV family. Once again, TRIM24 was identified as the non-class IV BRD with most affinity (IC50=1,600 nM), but with a 500-fold window of selectivity.
Selectivity against non-family targets:
NI-57 was screened for off-target pharmacology against a panel of 55 representative GPCRs, ion channels and enzymes (Eurofins, ExpressProfile) and was found to have minimal inhibition (<40 % at 10 uM).