摘要
目的评价布美他尼杂质N-亚硝基布美他尼的体内致突变风险和肝细胞DNA损伤风险。方法SD大鼠随机分为6组,分别为:溶媒对照组(溶媒为0.5%羧甲基纤维素钠),N-亚硝胺布美他尼100、300、1000 mg·kg^(-1)剂量组,阳性对照组1[N-乙基-N-亚硝基脲(ENU),40 mg·kg^(-1)]、阳性对照组2[甲磺酸乙酯(EMS),200 mg·kg^(-1)]。2个阳性对照组均为每组6只动物,其余每组12只动物。大鼠连续14 d经口灌胃给予受试物N-亚硝胺布美他尼。首次给药后约14 d和约28 d后采集外周血用于Pig-a基因突变率检测,末次给药后约3 h取肝细胞开展彗星试验。结果试验期间给予N-亚硝基布美他尼未导致异常临床症状和动物体重及摄食量改变。100、300、1000 mg·kg^(-1)剂量组动物肝细胞平均tail%DNA值(平均数/中位数)分别为2.90±0.38/2.34±0.46、3.58±0.27/2.87±0.51、3.45±0.59/2.21±1.44,与溶媒对照组相应数值相比未见差异,且无明显剂量效应相关性。首次给药后14 d,100、300、1000 mg·kg^(-1)剂量组动物平均RBC^(CD59-)和RET^(CD59-)百万分之发生率(RBC^(CD59-)百万分之发生率/RET^(CD59-)百万分之发生率)分别为2.9±1.6/1.2±0.8、2.8±2.4/1.3±1.5、2.4±1.0/1.3±1.5;首次给药后28 d,100、300、1000 mg·kg^(-1)剂量组动物RBC^(CD59-)百万分之发生率/RET^(CD59-)百万分之发生率分别为5.1±1.5/2.2±0.6、4.3±1.5/3.5±3.6、4.8±2.4/2.5±2.7。上述数值与相同时间点溶媒对照组相比均未见差异,且经统计学分析未见剂量效应相关性。结论连续经口给予N-亚硝基布美他尼14 d,SD大鼠的最大耐受量高于1000 mg·kg^(-1),未检出外周血基因突变风险和肝细胞DNA损伤风险。研究数据可为药品中遗传毒性杂质的合理监管和含N-亚硝基杂质药物的安全使用提供数据支持。
Objective To evaluate the mutagenicity risk and hepatocyte DNA damage risk of bumetanide impurity-N-nitrosobumetanil in vivo.Methods SD rats were randomly divided into six groups:control group(0.5%CMC-Na),100mg·kg^(-1)dose group,300 mg·kg^(-1)dose group,1000 mg·kg^(-1)dose group,positive control group 1(N-ethyln-nitrosourea,ENU,40 mg·kg^(-1))and positive control group 2(ethyl mesylate,EMS,200 mg·kg^(-1)).Both positive control groups consisted of six animals and the other groups had twelve animals in each.Rats were administered with N-nitrosobumetanil by oral intragastric administration for fourteen days.Peripheral blood was collected at about 14 days and 28 days after the initial administration for the detection of Pig-a gene mutation rates.Liver cells were collected about 3 hours after the last administration for comet test.Results During the study,N-nitrosobumetanil did not result in abnormal clinical symptoms and there was no change in animal body mass or food intake.The averaged tail%DNA values(mean/median)of animal hepatocytes in 100 mg·kg^(-1),300 mg·kg^(-1)and 1000 mg·kg^(-1)dosage groups were 2.90±0.38/2.34±0.46,3.58±0.27/2.87±0.51and 3.45±0.59/2.21±1.44,respectively,which showed no difference compared with the solvent control group,and no significant dose-effect correlation was observed.Fourteen days after the initial administration,the mean incidence of RBC^(CD59-)and RET^(CD59-)per million cells(RBC^(CD59-)/RET^(CD59-))in 100 mg·kg^(-1)dose group,300 mg·kg^(-1)dose group and 1000mg·kg^(-1)dose group were 2.9±1.6/1.2±0.8,2.8±2.4/1.3±1.5 and 2.4±1.0/1.3±1,respectively.Twenty-eight days after the initial administration,the mean incidence of RBC^(CD59-)and RET^(CD59-)per million cell(RBC^(CD59-)/RET^(CD59-))in the 100 mg·kg^(-1)dose group,300 mg·kg^(-1)dose group and 1000 mg·kg^(-1)dose group was 5.1±1.5/2.2±0.6,4.3±1.5/3.5±3.6,and 4.8±2.4/2.5±2.7,respectively.The above values were not different from those of the solvent control group at the same time point,and no correlation of dose effect was observed by statistical analysis.Conclusion After fourteen days of oral administration of N-nitrosobumetanilis,the maximum tolerance of SD rats exceeds1000 mg·kg^(-1),and the risk of peripheral blood gene mutation and hepatocyte DNA damage is not detected.The data can be used to back up the regulation of genotoxic impurities in drugs and the safe use of drugs containing N-nitroso impurities.
作者
文海若
黄勤
韩素芹
姜华
秦超
石皓琨
赵婷婷
耿兴超
汪祺
WEN Hairuo;HUANG Qin;HAN Suqin;JIANG Hua;QIN Chao;SHI Haokun;ZHAO Tingting;GENG Xingchao;WANG Qi(Beijing Key Laboratory,National Center for Safety Evaluation of Drugs,National Institutes for Food and Drug Control,Key Laboratory of Beijing for Nonclinical Safety Evaluation Research of Drugs,Beijing 100176,China;Guilin Pharmaceutical Co.,Ltd.,Guilin Guangxi 541004,China;Institute of Chinese Traditional Medicine and Ethnic Medicine,National Institutes for Food and Drug Control,Beijing 100050,China)
出处
《中国药物警戒》
2024年第3期307-312,318,共7页
Chinese Journal of Pharmacovigilance
基金
国家自然科学基金资助项目(82374033)。
