TrxR2-IN-1

TrxR2-IN-1 is an inhibitor of thioredoxin reductase 2 (TrxR2) with an IC50 value of 0.83 μM. It accumulates in mitochondria, disrupting their function and membrane potential, elevating reactive oxygen species (ROS) levels, and activating ASK1-mediated caspase-dependent apoptosis. TrxR2-IN-1 induces G2/M phase cell cycle arrest and inhibits cancer cell migration, making it applicable for hepatocellular carcinoma research.

TrxR2-IN-1 (Compound 23d) demonstrates potent inhibition of A549, HCT116, HeLa, Huh7, and MDA-MB-231 cancer cell viability, while showing high selectivity for normal HHL5 liver cells. In Huh7 cells, it exhibits an IC50 of 0.19 μM with a selectivity index (SI) of 15.24. TrxR2-IN-1 (0.8-1.6 μM; 24 h) induces dose-dependent morphological changes in Huh7 cells, indicating inhibited growth and cell death, and also hinders their migration. A dose-dependent G2/M phase arrest is observed in Huh7 cells with TrxR2-IN-1 (0.4-1.6 μM; 24 h). Antitumor agent-215 (0.4-1.6 μM; 24 h) induces apoptosis in a dose-dependent manner, disrupts mitochondrial membrane potential, and increases total reactive oxygen species (ROS) and mitochondrial superoxide levels in Huh7 cells. TrxR2-IN-1 (1.6 μM; 24 h) accumulates in the mitochondria of Huh7 cells at levels 2.6 times higher than its non-targeted precursor, 4d. Additionally, TrxR2-IN-1 (0.4-1.6 μM; 24 h) activates the ASK1-mediated caspase apoptotic pathway, enhancing levels of phosphorylated ASK1, cleaved caspase-9, and cleaved caspase-3 in a dose-dependent fashion.

TrxR2-IN-1 (Compound 23d) (1.25-5 mg/kg; subcutaneous injection; administered every 2 days for 14 days) exhibits potent and dose-dependent antitumor activity in the Huh7 liver cancer xenograft model.