The study developed humanized anti-CD33 CAR-T cells and antibody-drug conjugates (ADCs) for the targeted treatment of acute myeloid leukemia (AML), where CD33 is a highly expressed protein on AML cells. Antibodies were generated by immunizing mice and humanized, then used to construct CAR-T cells and ADCs with cytotoxic MMAE. In vitro experiments showed strong cytotoxic effects of clones Clone2HM, Clone3HM, Clone5HM, Clone6HM-Lu and Clone7 against Molm1 cells, with Clone7 outperforming the Gemtuzumab-MMAE control. In in vivo mouse models with Molm13-luciferase tumor cells, Clone3HM-MMAE treatment significantly reduced tumor signals to almost zero and prolonged survival compared to other groups. The body weight of mice remained stable during treatment, indicating a good safety profile. The results support further development of these therapies for AML.