Fig. 7.
On-target inhibition of FASN by compound 41. (A) Treatment of BT-474 lipogenic, HER2+ BC cells with compound 41 (4 hours, 50 μM) directly inhibited FASN activity, as shown by significant accumulation of the committed substrate for de novo lipogenesis, malonyl-CoA (*P < 0.05, one-way ANOVA; biologic duplicate and technical triplicate samples). (B) The ability of compounds 1 and 41 to specifically target the thioesterase domain of FASN was assessed using a FASN-TE substrate hydrolysis assay (N = 3 experiments). Under these conditions, the FASN-TE IC50 value of compound 1 was 1.47 ± 0.17 μM with a Hill slope of 0.773, whereas that of compound 41 was 14.58 ± 1.16 μM with a Hill slope of 0.824.