The article discusses the role of DNA topoisomerase II (Top2) in cancer chemotherapy and the development of anti-cancer drugs targeting this enzyme. Top2 plays crucial roles in DNA replication, transcription, and chromosome segregation, and its inhibition can generate DNA damage, making it an effective strategy for cancer treatment. The review highlights the distinction between Top2 poisons, which increase the levels of Top2:DNA covalent complexes, and catalytic inhibitors, which do not. Top2 poisons, such as etoposide and doxorubicin, generate DNA strand breaks and protein covalently bound to DNA, leading to cell death. Catalytic inhibitors, like bisdioxopiperazines, block Top2 catalytic activity without increasing covalent complexes. The review also explores the mechanisms of DNA damage repair by Top2, including nucleolytic excision and proteolytic degradation, and the potential of using these pathways to enhance therapeutic efficacy. Additionally, it discusses the challenges and future directions in Top2 targeting therapy, including the need for new drugs, the importance of isotype-specific targeting, and the potential of catalytic inhibitors as anti-cancer agents.The article discusses the role of DNA topoisomerase II (Top2) in cancer chemotherapy and the development of anti-cancer drugs targeting this enzyme. Top2 plays crucial roles in DNA replication, transcription, and chromosome segregation, and its inhibition can generate DNA damage, making it an effective strategy for cancer treatment. The review highlights the distinction between Top2 poisons, which increase the levels of Top2:DNA covalent complexes, and catalytic inhibitors, which do not. Top2 poisons, such as etoposide and doxorubicin, generate DNA strand breaks and protein covalently bound to DNA, leading to cell death. Catalytic inhibitors, like bisdioxopiperazines, block Top2 catalytic activity without increasing covalent complexes. The review also explores the mechanisms of DNA damage repair by Top2, including nucleolytic excision and proteolytic degradation, and the potential of using these pathways to enhance therapeutic efficacy. Additionally, it discusses the challenges and future directions in Top2 targeting therapy, including the need for new drugs, the importance of isotype-specific targeting, and the potential of catalytic inhibitors as anti-cancer agents.