ATOX1-IN-1

ATOX1-IN-1 is an inhibitor of ATOX1 (a copper chaperone protein) with a Kd of 12.5 μM. It induces intracellular copper accumulation, increases DNA methylation in the NOTCH1 promoter region, and inhibits the NOTCH1/HES1 signaling pathway. ATOX1-IN-1 enhances the sensitivity of liver cancer cells to Cisplatin and is applicable to liver cancer-related research.

ATOX1-IN-1 (Compound 8) effectively inhibits the viability of HepG2 and Huh7 liver cancer cells, with IC50 values of 16.5 μM and 22.0 μM respectively, showing higher selectivity for cancer cells over immortalized liver cells THLE-2. When combined with Cisplatin at a concentration of 10 μM, ATOX1-IN-1 (10 μM) displays enhanced antiproliferative and pro-apoptotic effects on HepG2 and Huh7 cells. Additionally, ATOX1-IN-1 at concentrations ranging from 5-15 μM can inhibit the NOTCH1/HES1 signaling pathway in these cells in vitro, leading to reduced mRNA and protein expression levels of NOTCH1 and HES1 in a concentration-dependent manner. This compound also increases intracellular copper levels in a dose-dependent fashion and enhances DNA methylation of the NOTCH1 promoter in HepG2 and Huh7 cells.

ATOX1-IN-1, administered intraperitoneally at 10 mg/kg once daily for 10 days, demonstrates antitumor activity in a HepG2 liver cancer xenograft model. When used in combination with Cisplatin, it results in a synergistic and well-tolerated antitumor effect.