N-亚硝基布美他尼SD大鼠体内致突变性风险研究  

Mutagenicity risk of N-nitrosobumetanil in SD rats

作　　者：文海若 黄勤 韩素芹 姜华[1] 秦超 石皓琨 赵婷婷 耿兴超[1] 汪祺[3] 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)

机构地区：[1]中国食品药品检定研究院国家药物安全评价监测中心,药物非临床安全评价研究北京市重点实验室,北京100176 [2]桂林南药股份有限公司,广西桂林541004 [3]中国食品药品检定研究院中药民族药检定所,北京100050

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)。

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 correlatio

关 键 词：N-亚硝基布美他尼 致突变性 遗传毒性 SD大鼠 Pig-a基因突变试验 彗星试验 灌胃 

分 类 号：R965[医药卫生—药理学]