Fig.9

Biological activity validation for WDR5 hit compounds from Pocket Crafter. A Chemical structures of the three compounds showing IC50 < 50 μM in HTRF assay, named PC-1, PC-2 and PC-3, B Dose response curves fitted from HTRF assay. IC50 = 35.6 ± 0.6, 27.5 ± 3.9 and 28.5 ± 0.2 μM respectively for compounds PC-1, PC-2, and PC-3 in the binding assay and all IC50s > 75 μM in the counter assay, C Melting peaks in WDR5 DSF by compound treatment. Thermal (Tm) shift: -4.2 °C by PC-1 and − 1.2 °C by PC-2 compared to the DMSO control. WM-662 was tested together as an assay control showing Tm shift of 3.2 °C, D Binding modes proposed by Pocket Crafter for the three hit compounds in WDR5 (yellow ribbon). PC-1 (Magenta), PC-2 (Orange) and PC3 (Cyan), E The comparison of binding modes among PC-1 (Magenta), WM-662 (Yellow), Compound 1 (red) and the Fesik lab’s Compound 12 (blue)