Figure 5 | Scientific Reports

Figure 5

From: Discovering cell-active BCL6 inhibitors: effectively combining biochemical HTS with multiple biophysical techniques, X-ray crystallography and cell-based assays

Figure 5The alternative text for this image may have been generated using AI.

TR-FRET assay development and correlation with the FP assay. (A) Schematic representation of the BCL6 BTB – BCOR peptide interaction as measured by TR-FRET assay format. (B) Unlabelled BCOR peptide competed with the Alexa-633 conjugated BCOR peptide in the TR-FRET reaction with an IC50 of 8 ± 0.8 µM, n = 20. (C) The two most potent HTS hits compound 17 and 21 showed respective IC50 values of 54 and 70 µM in the TR-FRET assay. (D) Correlation between the TR-FRET (x-axis) and the FP (y-axis) assays for the benzimidazolone series of BCL6 inhibitors. The BCL6 BTB concentration was 3 µM in the FP and 10 nM in the TR-FRET assay. The two biochemical assays correlated with a R2 of 0.82, but most compounds displayed higher potencies in the TR-FRET assay compared to the FP assay (red dotted trendline above the black dotted unity line). As compound potencies approached the tight binding limit of the FP assay (IC50 < 5 µM), the correlation between the two assays decreased.

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