Table 3:

Panel of small molecules tested in PrOF NMR, SPR, and AlphaScreen assays.

Ligand	PrOF NMR Kd (μM)	BPTF SPR Kd (μM)	19F BPTF SPR Kd (μM)	BPTF AlphaScreen IC50 (μM)
4	175	230 ± 50	280 ± 70	200a
5	*	NSB	NSB	36a
6	7†	41 ± 9	26 ± 10	19b
7	*	25 ± 10	19 ± 13	31b
8	20†	6 ± 2	3 ± 1	10 ± 2b
9	330†	Bound to reference	Bound to reference	NB

^*indicates molecules entered intermediate exchange and K_d values could not be determined

^†represents molecules with broadened resonances or a small dynamic range making quantification less accurate.

^‡indicates compound was insoluble at high concentration. PrOF NMR K_d values were determined from a single titration. SPR K_d values were based on data collected in triplicate using His₉-tagged BPTF bromodomain. NSB indicates that molecules bound non-specifically and 1:1 binding isotherms could not be generated. Bound to reference indicates molecule bound to chip surface at high ligand concentrations and K_d could not be determined. AlphaScreen IC₅₀ values were reported as the average of technical duplicates except for 8 which is an average of three experimental replicates.

^aMolecules were tested in the presence of 0.5% DMSO.

^bMolecules were tested at 0.25% DMSO. NB indicates non-binding up to 250 μM of compound.