Abstract: Cancer is one of the major diseases that seriously threaten human life and health. Traditional cancer treatment methods include chemotherapy, radiotherapy, and various other methods, but they all have their own side effects or limitations. In particular, the development of multidrug resistance to chemotherapeutic agents is a major obstacle for cancer treatment. The rise of peptide therapy provides a new approach to cancer treatment, as peptides have become promising anticancer agents due to their targeting specificity, low toxicity, and high selectivity. This article provides a comprehensive review of the anticancer mechanisms, clinical applications, and research and development progress of anticancer peptides (ACPs), as well as the computer-aided design of ACPs, including peptide-protein interaction-based design and artificial intelligence-based design, aiming to provide some reference for the design and development of novel ACP drugs.