Extended Data Fig. 5. Characterisation of small molecules with enhanced potency.
a) Inhibition (average ± s.e.m., n = 3) of the androgen-induced full-length AR transcriptional activity by compounds shown in Fig. 4a. b) Selected regions of Tau-5* 1H,15 N BEST-TROSY spectra in the absence (gray) and presence of 1 mol equivalent of EPI-001 (blue) and 1aa (red). c) Helical propensities of Tau-5R2_R3 in its apo form (black) and in bound conformations obtained from simulations run in the presence of EPI-002 (blue) and 1aa (red) with an indication of the positions of helical motifs and aromatic residues in the sequence. The data was obtained from the 300 K REST2 MD simulations. Values are presented as averages ± statistical errors from block averaging. d) Populations of aromatic stacking contacts between aromatic side chains of Tau-5R2_R3 and aromatic rings of EPI-002 (blue) and 1aa (red) with an indication of the positions of helical motifs and aromatic residues. The data was obtained from the 300 K REST2 MD simulations. Values are presented as averages ± statistical errors from block averaging. e) ChromLogD values of compounds developed from 1aa scaffold reporting their hydrophobicity (n = 3). f) Comparison of EPI-002 (35 µM) and enzalutamide (ENZA, 5 µM) with the most potent compounds (5 µM) to block AR-V7 transcriptional activity (average ± s.e.m., n = 3). g, h) Lack of a dose-dependent inhibition of AR-V7 transcriptional activity for 1ab (g) and 1bb (g). LNCaP cells that ectopically expressed AR-V7 were co-transfected with a V7BS3-luciferase reporter gene construct and incubated with the indicated concentrations of the compounds (average ±s.e.m., n = 3). i, j) 1ae blocked the proliferation of both LNCaP cells in response to androgen and AR-V-driven proliferation of LNCaP95 cells whereas Enzalutamide (ENZA) blocked androgen-induced proliferation driven by full-length AR in LNCaP cells but had poor potency against AR-V-driven proliferation of LNCaP95 (LN95) cells (average ± s.e.m., n = 3).