Zhu, YingYingZhuKe, Kun-BinKun-BinKeXia, Zhong-KunZhong-KunXiaLi, Hong-JianHong-JianLiSu, RongRongSuDong, ChaoChaoDongZhou, Feng-MeiFeng-MeiZhouWang, LinLinWangChen, RongRongChenWu, Shi-guoShi-guoWuZhao, HuiHuiZhaoGu, PengPengGuProf. LEUNG Kwong SakWong, Man-HonMan-HonWongLu, ganggangLuZhang, Jian-yingJian-yingZhangJiang, Bing- HuaBing- HuaJiangQiu, Jian-GeJian-GeQiuShi, Xi-NanXi-NanShiLin, Marie, Chia-miMarie, Chia-miLin2023-02-202023-02-202021Molecular Medicine 27, 2021, vol. 151528-3658http://hdl.handle.net/20.500.11861/7388Open accessBackground Cyclin-dependent kinases 2/4/6 (CDK2/4/6) play critical roles in cell cycle progression, and their deregulations are hallmarks of hepatocellular carcinoma (HCC). Methods We used the combination of computational and experimental approaches to discover a CDK2/4/6 triple-inhibitor from FDA approved small-molecule drugs for the treatment of HCC. Results We identified vanoxerine dihydrochloride as a new CDK2/4/6 inhibitor, and a strong cytotoxicdrugin human HCC QGY7703 and Huh7 cells (IC50: 3.79 μM for QGY7703and 4.04 μM for Huh7 cells). In QGY7703 and Huh7 cells, vanoxerine dihydrochloride treatment caused G1-arrest, induced apoptosis, and reduced the expressions of CDK2/4/6, cyclin D/E, retinoblastoma protein (Rb), as well as the phosphorylation of CDK2/4/6 and Rb. Drug combination study indicated that vanoxerine dihydrochloride and 5-Fu produced synergistic cytotoxicity in vitro in Huh7 cells. Finally, in vivo study in BALB/C nude mice subcutaneously xenografted with Huh7 cells, vanoxerine dihydrochloride (40 mg/kg, i.p.) injection for 21 days produced significant anti-tumor activity (p < 0.05), which was comparable to that achieved by 5-Fu (10 mg/kg, i.p.), with the combination treatment resulted in synergistic effect. Immunohistochemistry staining of the tumor tissues also revealed significantly reduced expressions of Rb and CDK2/4/6in vanoxerinedihydrochloride treatment group. Conclusions The present study isthe first report identifying a new CDK2/4/6 triple inhibitor vanoxerine dihydrochloride, and demonstrated that this drug represents a novel therapeutic strategy for HCC treatment.enCyclin-Dependent Kinases 2/4/6Hepatocellular CarcinomaVanoxerine DihydrochlorideTriple InhibitorDrug CombinationPeer Reviewed Journal Article10.1186/s10020-021-00269-4