Synthesis and Characterization of TH-2-31

RSL3, IKE were synthesized by the Stockwell lab^{23 (link),29 (link)}. All other chemicals were purchased from qualified vendors: staurosporine, Selleck Chemicals S1421; Z-VAD-FMK, Abcam ab120382; LCL-161, ChemieTek CT-LCL161; TNFα, PeproTech 300-01 A; ferric ammonium citrate, Sigma-Aldrich F5879; hemoglobin, Sigma-Aldrich H7379; ferrostatin-1, Sigma-Aldrich SML0583; Liproxstatin-1, Selleck Chemicals S7699; necrostatin-1s, Cell Signaling Technology 17802 S. TH-2-31 (N²,N³-dicyclohexyl-5-(3-methyl-1,2,4-oxadiazol-5-yl)pyridine-2,3-diamine) was synthesized by Curia. The structure of Th-2-31 was confirmed by NMR: ¹H NMR (400 MHz, Chloroform-d) δ 7.81 (d, J = 1.7 Hz, 1H), 6.86 (d, J = 1.9 Hz, 1H), 4.04–3.78 (m, 1H), 3.19 (td, J = 10.0, 4.2 Hz, 1H), 2.44 (s, 3H), 2.01 (d, J = 10.6 Hz, 4H), 1.89 – 1.64 (m, 4H), 1.58 (d, J = 13.0 Hz, 1H), 1.47 – 1.01 (m, 9H).