OXPHOS-IN-2 is an orally active inhibitor of OXPHOS, demonstrating potent inhibitory effects in PC9 cells (IC50= 12.3 nM) and Bxpc-3 cells (IC50= 250 nM in glucose medium, IC50= 17.5 nM in galactose medium). It modulates the NADH/NAD+ ratio and decreases ATP levels. OXPHOS-IN-2 induces apoptosis in tumor cells by activating reactive oxygen species (ROS) and downregulating Nrf2 levels. This compound is applicable for research in cancers such as non-small cell lung cancer and pancreatic cancer.

OXPHOS-IN-2 (Compound 28c) reduces the ratio of ATP to NAD+/NADH and inhibits AMPK phosphorylation in PC9 cells at concentrations of 0-1000 μM over 12 hours. It significantly impairs colony formation in PC9 cells at 0-20 nM over 14 days. At concentrations of 0-1 μM, OXPHOS-IN-2 induces apoptosis through a dose-dependent increase in reactive oxygen species (ROS) levels and a decrease in mitochondrial membrane potential in PC9 and Bxpc-3 cells. Additionally, at 0-50 nM, it causes DNA damage and elevates γ-H2AX expression in PC9 cells. OXPHOS-IN-2 inhibits the proliferation of various tumor cell lines in both glucose and galactose media, with IC50 values of 6.43 nM and 29.5 nM for H1975, 4.54 nM and 21.5 nM for H2228, 3.61 nM and 48.7 nM for KP-4, 5.78 nM and 29.8 nM for MKN45, 5.19 nM and 31.2 nM for HCT116, 1.31 nM and 7.5 nM for MDA-MB-231, 1.76 nM and 12.4 nM for MHCC-97H, and 5.10 nM and 35.1 nM for SiHA cells.

OXPHOS-IN-2 (Compound 28c) administered at 7.5 mg/kg orally once a day for 14 days significantly inhibits tumor growth in male BALB/c-nu PC9 tumor-bearing mice and demonstrates a favorable safety profile.