Abstract: Aim ：To evaluate the safety of glufosfamide in Beagle dogs. Methods ：The safety of glufosfamide in Beagle dogs following single iv and multiple iv administration for 3 months was observed. Results ：The minimum lethal dose of glufosfamide was 100 mg/kg.The maximum non-lethal dose of glufosfamide was 50 mg/kg.The approximate lethal dose of glufosfamide was 75-100 mg/kg.No serious adverse reactions was observed in Beagle dogs following multiple iv administration once a week for 3 months at the dose of 20 mg/kg，40 mg/kg，and 60 mg/kg.However，organ injury was observed extensively in the high dose-group and mediumt-dose group.The low-dose group showed a light change in individual organs. Conclusion ：Under the experimental conditions，40 mg/kg and 60 mg/kg iv administration once a week for 3 months can lead to extensive organ injury in Beagle dogs，so the dose shouldn′t be higher than 20 mg/kg.