中国药物警戒 ›› 2026, Vol. 23 ›› Issue (5): 487-493.
DOI: 10.19803/j.1672-8629.20260219

• 亚硝胺类药物相关杂质安全性研究专栏 • 上一篇    下一篇

杂环类亚硝胺杂质致突变性风险预测与试验验证

文海若1,2, 靳龙龙3△, 寇小旋1,2, 蒋晨晨1,2, 叶潇1,2, 张瑞秋1,2, 田冶1,2#, 吴先富3,*   

  1. 1中国食品药品检定研究院安全评价研究所,北京 100176;
    2药品监管科学全国重点实验室,北京 102629;
    3中国食品药品检定研究院标准物质和标准化管理中心,北京 102629
  • 收稿日期:2026-03-17 发布日期:2026-05-20
  • 通讯作者: *吴先富,男,博士,研究员,标准物质研究与药品质量控制。E-mail: wuxf99@163.com #为共同通信作者。
  • 作者简介:文海若,女,博士,研究员,遗传毒理学。△为并列第一作者。
  • 基金资助:
    国家自然科学基金资助项目（82473889）; 药品监管科学全国重点实验室课题“药品杂质遗传毒性评价新技术和生物标志物研究”（2023SKLDRS0128）

Mutagenic Risk Prediction and Experimental Validation of Heterocyclic Nitroso Impurities

WEN Hairuo1,2, JIN Longlong3△, KOU Xiaoxuan1,2, JIANG Chenchen1,2, YE Xiao1,2, ZHANG Ruiqiu1,2, TIAN Ye1,2#, WU Xianfu3,*   

  1. 1Institute of Safety Evaluation, National Institutes for Food and Drug Control, Beijing 100176, China;
    2State Key Laboratory of Drug Regulatory Science, Beijing 102629, China;
    3National Institutes for Food and Drug Control, Center of Reference Standards and Standardization, Beijing 102629, China
  • Received:2026-03-17 Published:2026-05-20

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摘要： 目的 评估3个药物成分相关的杂环类亚硝胺杂质致突变性风险,为确定其人体可接受摄入量提供参考。方法 分别采用Lhasa、Leadscope和CASE Ultra 3个构效关系（Structure-Activity Relationship, SAR）预测软件对N-亚硝基帕罗西汀、N-亚硝基瑞波西汀和N-亚硝基氢溴酸伏硫西汀的致突变风险进行预测;在无代谢活化和30%仓鼠S9代谢活化条件下开展包括鼠伤寒沙门氏菌TA98、TA100、TA1535、TA1537和大肠杆菌 WP2 uvrA (pkM101) 的增强细菌回复突变试验,评价其致突变性风险;使用基于R语言程序包的PROAST软件比较杂环类亚硝胺杂质的基准剂量（Benchmark Dose,BMD）水平,比较致突变性风险强弱与结构修饰之间的关联。结果 构效预测显示,Lhasa和Leadscope基于结构特点认为3种杂环类亚硝胺杂质均存在致突变性风险;而CASE Ultra则基于企业内部研究数据,判断N-亚硝基帕罗西汀不存在致细菌回复突变风险。增强细菌回复突变试验中,在非代谢活化条件下,所有受试物结果均为阴性;在仓鼠S9代谢活化条件下,除N-亚硝基帕罗西汀外,其他杂环类亚硝胺杂质均可诱导细菌回复突变数增加,存在致突变性风险。基于基准剂量可信下限（Benchmark Dose Lower Confidence Limit, BMDL）数值认为N-亚硝基瑞波西汀对TA98的致突变效力最强、N-亚硝基氢溴酸伏硫西汀对TA100的致突变效力最强。结论 3种常见的杂环类亚硝胺杂质除N-亚硝基帕罗西汀外,其余均有细菌致突变性,并发现含六元环内亚硝基的亚硝胺杂质易导致碱基移码突变,上述研究结果为杂环类亚硝胺杂质致突变性风险评价和基于化学结构活性关系的亚硝胺杂质风险预测提供参考。

关键词: 杂环类亚硝胺杂质, 致突变性, 构效关系预测, 增强细菌回复突变试验, 基准剂量水平

Abstract: Objective To evaluate the mutagenic risk of three drug-related heterocyclic nitroso impurities, and to provide data for setting regulatory limits. Methods Three (quantitative) structure-activity relationship [(Q)SAR] prediction software packages, namely Lhasa, Leadscope, and CASE Ultra, were used to predict the mutagenic risk of N-nitroso-paroxetine, N-nitroso-reboxetine, and N-nitroso-vortioxetine hydrobromide. Enhanced bacterial reverse mutation tests were conducted, involving Salmonella typhimurium strains TA98, TA100, TA1535, TA1537 and Escherichia coli strain WP2 uvrA (pKM101), both in the absence and presence of a 30% hamster S9 metabolic activation kit to evaluate their mutagenic potential. The benchmark dose (BMD) levels of these heterocyclic nitroso impurities were compared using the PROAST software (an R-based package) to study the correlations between the strength of mutagenic risk and structural modifications. Results As was predicted by (Q)SAR, Lhasa and Leadscope, based on structural characteristics, suggested a mutagenic risk for all the three heterocyclic nitroso impurities. In contrast, CASE Ultra, which relied on internal research data alone, predicted that N-nitroso-paroxetine posed no risk of bacterial reverse mutation. In the enhanced bacterial reverse mutation tests, all the test substances yielded negative results under non-metabolic activation. However, under metabolic activation with hamster S9, all the heterocyclic N-nitroso impurities, except N-nitroso-paroxetine, induced an increase in the number of revertant colonies, indicating a mutagenic risk. Based on BMDL values, N-nitroso-reboxetine was believed to be the most mutagenically potent for strain TA98, and N-nitroso-vortioxetine hydrobromide for strain TA100. Conclusion The three common heterocyclic nitroso impurities, except N-nitroso-paroxetine, demonstrate bacterial mutagenicity. Nitrosamine impurities containing a nitroso group within a six-membered ring are likely to induce frameshift mutations. These findings are expected to provide data for regulatory control of impurities in heterocyclic drugs and for risk prediction of nitrosamine impurities based on chemical structure-activity relationships.

Key words: Heterocyclic Nitroso Impurities, Mutagenicity, Structure-Activity Relationship Prediction, Enhanced Bacterial Reverse Mutation Test, Benchmark Dose Level

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