CHNQD-01522 is a microtubule inhibitor that targets the colchicine binding site on β-tubulin. It disrupts microtubule polymerization and can evade P-glycoprotein transport in cancer cells. This compound inhibits cancer cell proliferation, prevents tumor cell colony formation, arrests the cell cycle at the G2/M phase, and induces apoptosis. CHNQD-01522 upregulates Bax and activates caspase-9 and caspase-3. In xenograft models of subcutaneous and orthotopic liver cancer, it exhibits antitumor activity and is applicable for hepatocellular carcinoma research.