GPX4-IN-20 is a derivative of Arctigenin and acts as a molecular glue degrader of GPX4, inducing ferroptosis by increasing lipid ROS levels and inhibiting GSH levels. It reduces the protein expression and enzyme activity of GPX4 in a dose-dependent manner without affecting other ferroptosis-related proteins. GPX4-IN-20 promotes ubiquitin-dependent proteasomal degradation of GPX4 and elevates malondialdehyde (MDA) levels in HCT-116 cells. This compound is applicable in colorectal cancer research.

GPX4-IN-20 (compound W25) exhibits cytotoxic activity in HCT-116 cells at concentrations of 2.5-10 μM over periods ranging from 24 to 72 hours. It suppresses both expression and activity of GPX4 protein, facilitating ubiquitin-dependent proteasomal degradation, without significantly affecting other ferroptosis regulators such as FSP1, FTH1, NCOA4, and SLC7A11. The compound does not alter GPX4 mRNA levels but inhibits clonogenic formation in HCT-116 cells in a dose-dependent manner when applied at 2.5-10 μM for 6 hours. In HT-29 cells, GPX4-IN-20 impedes growth with an IC50 of 4.60 μM after 24 hours. It significantly elevates lipid ROS levels in a dose- and time-dependent fashion and decreases GPX4 levels, while inducing a dose-dependent rise in intracellular MDA content in HCT-116 cells. Finally, GPX4-IN-20 binds to GPX4 at 10 μM for 24 hours, altering its thermostability.