逃离关注人群:体外实验证据支持 N-亚硝基-氢氯噻嗪的非突变性

IF 4.8 2区 医学 Q1 TOXICOLOGY
R. D. Gandhi, S. Hickert, Y. Hoevelmann, C. D. Mee, J. Schlingemann, A. Adams, A. Blanazs, S. Simon, J. Elloway, L. Rigger, A. Teasdale, C. V. Beaumont, L. Wright, A. Doherty
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引用次数: 0

摘要

近年来,药品中的亚硝胺杂质受到了监管部门的严格审查,亚硝胺药物物质相关杂质(NDSRI)被视为一组关注杂质,而不管预测的致突变潜力如何。在这里,我们描述了一项关于 NDSRI N-亚硝基-氢氯噻嗪(NO-HCTZ)的案例研究,该物质在细菌反向突变(艾姆斯)试验中呈阳性,但在试验条件下不稳定,会产生甲醛等产物。NO-HCTZ 的致突变特征与亚硝胺的致突变特征不一致,在没有新陈代谢活化的情况下表现出致突变性,而与甲醛的致突变特征非常一致。为了进一步评估,开发了一种改进的阿姆斯系统,其中包括谷胱甘肽(3.3 毫克/板)来清除甲醛。所用菌株为伤寒杆菌 TA98、TA100、TA1535 和 TA1537 以及大肠杆菌 WP2 uvrA/pKM101。在这一系统中,甲醛含量大大降低,S-(羟甲基)谷胱甘肽(谷胱甘肽与甲醛之间形成的加合物)的含量也随之增加。在改良系统(1.6-5000 微克/板)中重新测试 NO-HCTZ 后,在原始系统中 NO-HCTZ 具有诱变作用的菌株(TA98、TA100 和 WP2 uvrA/pKM101)中观察到诱变反应明显降低,这表明是甲醛而不是 NO-HCTZ 驱动了诱变反应。在菌株 TA1535 中,改良系统中观察到返祖菌落增加,这可能是由于 NO-HCTZ 在艾姆斯试验条件下形成了噻嗪降解产物。总之,这些数据支持NO-HCTZ的非致畸性命名,并证明了当阿姆斯试验的阳性结果与预期结果不一致时,进一步调查的价值。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Escaping the cohort of concern: in vitro experimental evidence supports non-mutagenicity of N-nitroso-hydrochlorothiazide

Escaping the cohort of concern: in vitro experimental evidence supports non-mutagenicity of N-nitroso-hydrochlorothiazide

In recent years, nitrosamine impurities in pharmaceuticals have been subject to intense regulatory scrutiny, with nitrosamine drug substance-related impurities (NDSRIs) treated as cohort of concern impurities, regardless of predicted mutagenic potential. Here, we describe a case study of the NDSRI N-nitroso-hydrochlorothiazide (NO-HCTZ), which was positive in the bacterial reverse mutation (Ames) test but is unstable under the test conditions, generating formaldehyde among other products. The mutagenic profile of NO-HCTZ was inconsistent with that expected of a mutagenic nitrosamine, exhibiting mutagenicity in the absence of metabolic activation, and instead aligned well with that of formaldehyde. To assess further, a modified Ames system including glutathione (3.3 mg/plate) to remove formaldehyde was developed. Strains used were S. typhimurium TA98, TA100, TA1535, and TA1537, and E. coli WP2 uvrA/pKM101. In this system, formaldehyde levels were considerably lower, with a concomitant increase in levels of S-(hydroxymethyl)glutathione (the adduct formed between glutathione and formaldehyde). Upon retesting NO-HCTZ in the modified system (1.6–5000 µg/plate), a clear decrease in the mutagenic response was observed in the strains in which NO-HCTZ was mutagenic in the original system (TA98, TA100, and WP2 uvrA/pKM101), indicating that formaldehyde drives the response, not NO-HCTZ. In strain TA1535, an increase in revertant colonies was observed in the modified system, likely due to a thiatriazine degradation product formed from NO-HCTZ under Ames test conditions. Overall, these data support a non-mutagenic designation for NO-HCTZ and demonstrate the value of further investigation when a positive Ames result does not align with the expected profile.

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来源期刊
Archives of Toxicology
Archives of Toxicology 医学-毒理学
CiteScore
11.60
自引率
4.90%
发文量
218
审稿时长
1.5 months
期刊介绍: Archives of Toxicology provides up-to-date information on the latest advances in toxicology. The journal places particular emphasis on studies relating to defined effects of chemicals and mechanisms of toxicity, including toxic activities at the molecular level, in humans and experimental animals. Coverage includes new insights into analysis and toxicokinetics and into forensic toxicology. Review articles of general interest to toxicologists are an additional important feature of the journal.
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