Fig. 1: Zharp1-163 is a potent dual inhibitor of ferroptosis and necroptosis.
A Schematic representation of the screening process for dual inhibitors of ferroptosis (Survival exceeds 50%) and necroptosis (Survival exceeds 50%). B Chemical structure of Zharp1-163. C HT-1080 cells were pretreated with the indicated concentrations of Zharp1-163 for 2 h and then treated with erastin or RSL3 for 16 h. Cell viability was assessed by measuring ATP levels. D MEFs were pretreated with the indicated concentrations of Zharp1-163 for 2 h and then treated with erastin or RSL3 for 16 h. Cell viability was assessed by measuring ATP levels. E Dose–response curve and EC50 values for the effects of Zharp1-163 on TNF-induced necroptosis in HT-29 cells. The cells were pretreated with the indicated concentrations of Zharp1-163 for 2 h, followed by treatment with T (40 ng/ml), S (100 nM) and Z (20 μM) for 16 h. Cell viability was assessed by measuring ATP levels. F MEFs were pretreated with the indicated concentrations of Zharp1-163 for 2 h, followed by treatment with T (40 ng/ml), S (100 nM) and Z (20 μM) for 12 h. Cell viability was assessed by measuring ATP levels. G L929 cells were pretreated with the indicated concentrations of Zharp1-163 for 2 h, followed by treatment with T (40 ng/ml) and Z (20 μM) for 12 h. Cell viability was assessed by measuring ATP levels. H MEFs were pretreated with the indicated concentrations of Zharp1-163 for 2 h prior to treatment with T (40 ng/ml) and S (100 nM) for 24 h. Cell viability was assessed by measuring ATP levels. I THP-1 cells were pretreated with the indicated concentrations of Zharp1-163 or 5 μM disulfiram (positive control) for 2 h and then treated with LPS (1 µg/ml) for 4 h, followed by nigericin (10 μM) for 2 h. Cell viability was assessed by measuring ATP levels. T, TNF-α; S, Smac mimetic; Z, z-VAD; L, lipopolysaccharide. The data are presented as the mean ± SD of triplicate samples. n.s., not significant; ****P < 0.0001.
