Figure 2.

(A) Temperature shift data measured on synthesized inhibitors as well as clinical BzDs. Synthesized compounds have been initially screened at 100 μM compound concentration and interacting compounds were re-evaluated at 10 μM compound concentration. Temperature shift data are color coded as indicated in the figure. (B) Example of a raw data trace of a temperature shift experiment. Shown are data measured on BRD4(1) alone (black line) and BRD4(1) in the presence of 10 mM alprazolam (red line). (C) Isothermal titration calorimetry data. Shown are data measured on alprazolam (red) and BzD-7 (black). The panel shows raw binding heats of 8 μL injections of BRD4(1) into a solution of each of the two inhibitors. The first injection (2 μL) was not included into the data analysis. The insert shows normalized binding heats and non-linear least squares fit to the experimental data (solid lines). The derived thermodynamic data are compiled in Table 1.