Topoisomerase I/II inhibitor 2 (compound 1a) can significantly reduce the xenograft tumor growth in mice model which can be used in treating liver cancer. Topoisomerase I/II inhibitor 2 is a DNA topoisomerase I/II dual inhibitor. Topoisomerase I/II inhibitor 2 is a potent Topoisomerase inhibitor with IC50 values of 6.83 μM and 9.82 μM for LM9 cells and Huh7 cells [1].

Topoisomerase I/II inhibitor 2 (compound 1a) demonstrates a range of beneficial activities against cancer cell lines, showing strong anti-proliferative effects at concentrations between 0-150 μM over 72 hours, particularly against two human hepatocellular carcinoma cell lines, HuH7 and LM9. At a lower dose of 20 μM for 24 hours, this compound does not cause DNA damage in HuH7 cells, though some damage is observed in LM9 cells. Its ability to inhibit cell proliferation in both LM9 and HuH7 cells is concentration-dependent, evident from treatments lasting 1-2 weeks with 1.25-8 μM and from blocking the cell cycle at the G2/M phase at 0-20 μM for 48 hours. Additionally, this inhibitor reduces the migration and invasion of these cells and downregulates the expression of matrix metalloproteinases-9 (MMP-9) in a concentration-dependent manner, with significant effects noticeable at dosages ranging from 0-20 μM over 24 to 48 hours. Notably, concentrations of 3.5-20 μM over 48 hours can damage mitochondrial function and induce apoptosis in these cells. Investigations into its anti-proliferative activity across various cell lines (including LM9, HuH7, SK-hep-1, HepG2, HT-29, HCT-116, RKO, SW480, MCF-7, MDA-B-231, HGC-27, SGC-7901, BGC-823, A549, U251, HL-60, LO2) confirmed its efficacy, especially in HuH7 and LM9. Further analyses through Western Blot and cell cycle assessments corroborate its effectiveness in inhibiting MMP-9 expression and arresting the cell cycle at the G2/M phase, respectively, thus highlighting its potential as a targeted therapy in human hepatocellular carcinoma treatment.