MDMX/MDM2-IN-2 is a potent dual inhibitor of p53-MDM2/MDMX, with dissociation constants (Kis) of 0.23 μM for MDM2 and 2.45 μM for MDMX. It disrupts the interaction between p53 and MDM2 proteins, reinstating p53 functionality to induce cell cycle arrest and apoptosis. Additionally, this compound impedes cell migration and invasion, exhibiting notable antitumor activity [1].

MDMX/MDM2-IN-2 exhibits moderate antiproliferative activity in HCT116 and SH-SY5Y cells with IC50 values of 0.68 μM and 0.54 μM, respectively, and demonstrates low cytotoxicity towards normal human bronchial epithelial BEAS-2B cells and LO2 hepatocytes with IC50 values of 17.96 μM and 15.93 μM, respectively [1]. The compound, at concentrations of 0.6-2.4 μM for 48 hours, induces apoptosis and causes cell cycle arrest at the G1 phase in both HCT116 and SH-SY5Y cells [1]. Additionally, MDMX/MDM2-IN-2 increases levels of p53 and its downstream targets MDM2, MDMX, p21, and cleaved-caspase3 [1]. At 0.4-0.8 μM, it significantly inhibits colony formation, migration, and invasion in these cell lines [1].