Topoisomerase I/II inhibitor 3 (compound 7) is a potent dual inhibitor of topoisomerase I (Topo I) and II (Topo II) . By inhibiting PI3K /Akt/mTOR signaling pathway, Topoisomerase I/II inhibitor 3 can inhibit cell proliferation, invasion and migration, and induce apoptosis . Topoisomerase I/II inhibitor 3 has research value in liver cancer.

Topoisomerase I/II inhibitor 3 (compound 7) (0-100 μM) can insert into DNA and change the topology of DNA, cause DNA damage to a certain extent [1]. Topoisomerase I/II inhibitor 3 (0-4 μM, 24 h) inhibits HCC (hepatocellular carcinoma) cells proliferation in a dose-dependent manner, inhibits migration and invasion of LM9 and HuH7 cells by inhibiting the expression of MMP-9 [1]. Topoisomerase I/II inhibitor 3 (0-14 μM, 48 h) significantly induces the apoptosis of LM9 and HuH7 cells, and induces mitochondrial dysfunction and ROS burst in a dose-dependent manner [1]. Topoisomerase I/II inhibitor 3 (0-7 μM, 48 h) reduces the expression of the inhibitory factor Bcl-2 and promotes the expression of mitochondria-dependent apoptosis-related proteins such as Bax, cytochrome C, cleaved-caspase-3 and cleaved-caspase-9; inhibits the phosphorylation of PI3K/Akt/mTOR signaling pathway [1]. Cell Proliferation Assay Cell Line: HCC cells (HuH7 and LM9) [1] Concentration: 0, 0.5, 1, 2, and 4 μM Incubation Time: 24 h Result: Inhibited HCC cells proliferation in a dose-dependent manner, with IC 50 values of 2.10 μM (LM9) and 1.93 μM (HuH7), respectively, and dose-dependently inhibited the formation of cell colonies. Apoptosis Analysis Cell Line: LM9 and HuH7 cells [1] Concentration: 0, 1.8, 3.5, 7, and 14 μM Incubation Time: 48 h Result: Significantly induced the apoptosis of LM9 and HuH7 cells in a concentration-dependent manner. Western Blot Analysis Cell Line: LM9 and HuH7 cells [1] Concentration: 0, 1.8, 3.5, and 7 μM Incubation Time: 48 h Result: Reduced the expression of the inhibitory factor Bcl-2 and promoted the expression of mitochondria-dependent apoptosis-related proteins such as Bax, cytochrome C, cleaved-caspase-3 and cleaved-caspase-9; inhibited the phosphorylation of PI3K/Akt/mTOR signaling pathway.

Topoisomerase I/II inhibitor 3 (compound 7) (male Kunming mice, 0-400 mg/kg, IP, once) causes the mice in the 400 mg/kg group die, with the LD 50 between 250 and 400 mg/kg [1]. Animal Model: Male Kunming mice (19-22 mg, 8 mice, 4 groups) [1] Dosage: 200, 250, 400 mg/kg (dissolved in 5% DMSO and castor oil) Administration: IP, once Result: Did not cause the mice in the 250 mg/kg and 200 mg/kg groups die after 2 weeks of administration, and caused the mice in the 400 mg/kg group die, with the LD 50 between 250 and 400 mg/kg.