NCA029 is a potent activator of human caseinolytic protease P (HsClpP) with an EC50 of 0.15 μM. It targets HsClpPP and triggers an ATF3-dependent integrated stress response, resulting in the death of colon cancer cells.

NCA029 inhibits human colorectal adenocarcinoma cell lines (COAD), with IC50 values for HCT116, SW620, HCT15, SW480, and DLD-1 recorded at 1.1 μM, 1.5 μM, 3.5 μM, 5.4 μM, and 3.1 μM, respectively. It increases reactive oxygen species (ROS) levels in HCT116 cells in a dose-dependent manner, indicating impaired respiratory chain activity. At concentrations of 0.5-2 μM for 48 hours, NCA029 suppresses the proliferation and metastasis of tumor cellsand induces apoptosis in a dose-dependent fashion while arresting HCT116 cells at the G2/M phase. Additionally, NCA029 (0.5-2 μM) reduces the expression of SDHB and increases the pro-apoptotic protein BAX while decreasing the expression of the anti-apoptotic protein Bcl-2 in a dose-dependent manner.

NCA029 (1.25-5 mg/kg; intravenous injection; twice weekly; for 21 days) significantly suppresses tumor growth in HCT116 xenografts.