那些含有基因毒性杂质的药物

对于药物研发而言，基因毒性杂质(又称遗传毒性杂质)已经不算是新鲜事物了。这类能直接或间接损伤细胞DNA，产生致突变和致癌作用的物质属于杂质研究中对于特定种类杂质要求的一个分支。

我国加入ICH以后，药品研发技术要求势必与国际接轨，对于基因毒性杂质的评估要求也必然与ICH成员国要求统一甚至趋于更加严格。这方面的体现之一便是2017年12月CFDA发布的《已上市化学仿制药(注射剂)一致性评价技术要求(征求意见稿)》。征求意见稿针对遗传毒性杂质指出：“根据相关文献、参比制剂的情况，通过对生产工艺、产品降解途径的分析，判断是否可能产生潜在的遗传毒性杂质，必要时进行针对性的研究，根据研究结果按照相关技术指导原则进行控制。”不仅对于注射剂再评价，在目前开展的口服固体制剂一致性评价、新药和仿制药申报的研究中，遗传毒性杂质的研究也越来越受到重视。换个层面来想，在药学研究中，对于杂质的研究和控制已经厮杀成红海，而作为杂质研究分支的基因毒性杂质研究可能会成为未来重要的竞争战场，对基因毒性杂质的轻视势必要付出惨重的代价。

鉴于此，笔者利用SciFinder Scholar数据库，检索并整理了国外针对特定药物品种基因毒性杂质的研究性英文文献(详见表1)。结果发现，基因毒性杂质的英文文献出现于2006年，并且在2006-2008年期间发表的文献主要集中为基因毒性杂质综述和探讨性文献。经过笔者深入了解，发现这主要是由于针对基因毒性杂质研究指南诞生背景所致。2006年EMA首先颁布了《基因毒性杂质限度指南》(Guideline on the Limits of Genotoxic Impurities)，并自2007年1月1日起正式实施。该指南为限制新活性物质中的基因毒性杂质提供了解决问题的框架和具体做法。EMEA之所以要发布这一指南，主要是因为在当时的指南中，对EMEA批准上市的新药中所含基因毒性杂质的可接受水平缺乏相应的监管条款。该指南是第一个直接针对基因毒性杂质的监管规定，重点关注的是在新药合成、纯化和储存运输过程中，最有可能产生的实际潜在性的基因毒性杂质。但该指南并不适用于当时之前已经获批上市的产品，除非当时某些产品存在让人担忧的特殊原因。基于EMEA上述指南实施两年后和ICH相关指南，FDA于2008年12月也正式签发了类似指南《Guidance for Industry-Genotoxic and Carcinogenic Impurities in Drug Substances and Products：Recommended Approaches》。对指南发布的背景、原料药和制剂中的基因毒性杂质生成的预防办法、基因毒性杂质的分析方法、处理方法和减少方法，上市申请和临床研究申请的可接受限度作了详细阐述和说明。但于2015年5月28日撤销了该指南，又陆续发布了M7《Assessment and Control of DNA Reactive (Mutagenic) Impurities in Pharmaceuticals to Limit Potential Carcinogenic Risk》和M7 (R1)《Addendum to ICH M7: Assessment and Controlof DNA Reactive (Mutagenic) Impurities in Pharmaceuticals to Limit Potential Carcinogenic Risk》。此外FDA还发布了草药及其制剂中基因毒性杂质评估指南。

综上可见，2006-2008年为国际上药品基因毒性杂质研究指南出台和认知接受阶段，所以这段时间主要是基因毒性杂质综述类文献较多，但这些文献可以更好的帮助我们对于背景加深认知和了解。2008开始有了针对特定药物品种基因毒性杂质的研究文献，而且直到2016年，呈现逐年递增的趋势。目前已有50余个特定药物品种的基因毒性杂质的研究文献(详见表1)。

此次CFDA推行的注射剂再评价中，要重新评估过去上市品种中的基因毒性杂质，这一举措已经超越了FDA和EMA的监管要求层次，同时这也为基因毒性杂质的考察带来了一定的困难，毕竟之前上市的老原研品种在上市时可能并未对基因毒性杂质进行过系统的研究，存在着无鉴可借的情况。因此本文从发表的国外英文文献入手，对相关基因毒性杂质研究性文献进行了简要的汇总，旨在提纲挈领，以鸟瞰的方式初步领略已有药物品种基因毒性杂质的研究进展。

最后，想强调一点，文中汇总的各个药物的基因毒性杂质是根据文中给出的杂质结构名称翻译而成，具体结构还需从对应的参考文献中去了解。笔者之所以还是给出中文翻译，而不是直接copy英文名称，主要是考虑给予大多数母语并不是英语的读者们一个直接的感性认识，这也是本文的目的所在。如果需要了解更为详细的具体信息，则可到对应的参考文献中去探寻。

对应的参考文献

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20. Trace level quantification of the(-)2-(2-amino-5-chlorophenyl)-4-cyclopropyl-1,1,1-trifluoro-3-butyn-2-ol genotoxic impurity in efavirenz drug substance and drug product using LC-MS/MS. Jaishetty, Nagadeep; Palanisamy, Kamaraj; Maruthapillai, Arthanareeswari; Jaishetty, Rajamanohar. Scientia Pharmaceutica (2016), 84(3), 456-466.

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23. Assessment and quantification of genotoxic impurities of Ziprasidone an antipsychoticdrug. Katta, Krishna; Kandula, Venkata Ramana; Rao, T. Siva; Jain, Kirti Kumar;Rao, D. Venugopala; Babu, J. Moses. Journal of Chemical and PharmaceuticalResearch (2015), 7(6), 739-750.

24. Determination of glycidol as a genotoxic impurity in linezolid drug substanceby gas chromatography. Kalgutkar, Ravi; Ramakrishna, K.; Shenoy, Shobha; Srinivasarao, V.. Analytical Chemistry: An Indian Journal (2015), 15(12),505-508.

25.Development and validation of RP-HPLC method for determination of genotoxicimpurity DSN-V in dasatinib. Rao, Kamala Govinda; Kumar, Konidala Sathish;Kumar, Mugada Vinod; Koralla, Sravani; Bommareddy, Nithin. European Journal of Biomedical and Pharmaceutical Sciences (2015), 2(4), 445-454.

26.Forced degradation study of lamivudine under the scope of genotoxic impurity. Prakash, Anuj; Teotia, Anil K.; Farooqi, Javed A.; Singh, G. N.. World Journal of Pharmacy and Pharmaceutical Sciences (2015), 4(6), 1032-1041.

27. Genotoxic impurity method development and validation by GC-MS for the analysis of methyl methane sulfonate (MMS) in Zidovudine drug substance. Kakasaheb,Nalavade Atul; Ramakrishna, K.; Srinivasarao, V.. Analytical Chemistry: AnIndian Journal (2015), 15(5), 184-189.

28. Method development and validation for quantitative determination of para chloroaniline, an impurity, in efavirenz active pharmaceutical ingredient by liquid chromatography - electrospray ionization - mass spectrometry/mass spectrometry. Rao, Dokku Raghava; Sreenivasulu, Vudagandla; Maheswari, B. N.Uma; Reddy, M. Veera Narayana; Kumar,Nadavala Siva; Krishnaiah, Abburi. Indian Journal of Advances in Chemical Science (2015), 3(3), 267-273.

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38. Trace level quantification of 1-(3-chloropropyl)-4-(3-chlorophenyl) piperazine HCl genotoxic impurity in trazodone using LC-MS/MS. Venugopal, Nandigam; Vijaya Bhaskar Reddy, Ambavaram; Madhavi, Gajulapalle; Jaafar, Jafariah; Madhavi, Vemula;Gangadhara Reddy, Kalluru. Arabian Journal of Chemistry (2014), Ahead of Print.

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49.Quantification of genotoxic impurity 4-Chloro-1-Hydroxy Butane Sulfonic Acid Sodium Salt by LCMS/MS in Sumatriptan Succinate. Narayana,M. B. V.; Chandrasekhar, K. B.; Rao, B. M.. Journal of Bioanalysis &Biomedicine (2012), 4(6), 104-107.

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51.Quantification of 4-Oxiranylmethoxy-9H-Carbazole agenotoxic impurity in Carvedilol drug substances by LC-MS. Rao, M. Srinivasa; Rao, Sumathi V.; Ray, U. K.; Kumar, G. Sri Siva; Sharma, Hemant Kumar;Mukkanti, K.. Journal of Bioanalysis & Biomedicine (2010), 2(4), 091-095.

52. Analytical control of genotoxic impurities in the pazopanib hydrochloride manufacturing process. Liu, David Q.; Chen, Ted K.;McGuire, Michael A.; Kord, Alireza S.. Journal of Pharmaceutical and BiomedicalAnalysis (2009), 50(2), 144-150.

53. Preparative Chromatography Technique in the Removal of Isostructural Genotoxic Impurity in Rizatriptan:Use of Physicochemical Descriptors of Solute and Adsorbent. Maddula, Srinivasula Reddy; Kharkar,Manoj; Manudhane, Kushal; Kale, Sandeep; Bhori, Abijar; Lali, Arvind; Dubey, P.K.; Sarma, K. R. Janardana; Bhattacharya, Apurba; Bandichhor, Rakeshwar.Organic Process Research & Development (2009), 13(4), 683-689.

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