GPX4-IN-14 (compound 2c) acts as a GPX4 inhibitor, exhibiting both free radical scavenging activity (with a maximum scavenging rate of 72.52%) and anti-tumor proliferation activity in vitro. This compound targets GPX4 protein, elevating lipid peroxide and intracellular Reactive Oxygen Species (ROS) levels, which induces ferroptosis and contributes to its anti-tumor proliferation effects.

GPX4-IN-14 (100 μM, 30 min) exhibits potent radical scavenging activity, achieving a maximal scavenging rate of 72.52%. It displays antiproliferative effects against several cancer cell lines including SW480, HCT116, HepG2, and MCF-7 with IC50 values of 5.61 μM, 6.59 μM, 18.23 μM, and 9.73 μM respectively, and has an IC50 of 11.64 μM in LO2 cells. At concentrations of 3.3, 6.6, and 13.2 μM over 24 hours, GPX4-IN-14 inhibits the GPX4 pathway in HCT116 cells, inducing a dose-dependent accumulation of intracellular ROS and lipid ROS, which leads to ferroptosis and mitochondrial membrane potential suppression. Additionally, GPX4-IN-14 increases intracellular iron levels in a dose-dependent manner within HCT116 cells at the same concentrations and duration.