目的:通过中间体检测的控制策略,建立甲磺酸伏美替尼关键中间体N2-(2-(二甲胺基)乙基-N2-甲基-N5-(4-(1-甲基-1H-吲哚-3-基)嘧啶-2-基)-6-(2,2,2-三氟乙氧基)吡啶-2,3,5-三胺(AST 2815)质量标准中氯甲烷、氯乙烷、乙酰胺、4-氯-1-丁醇4个基因毒性杂质的检测方法,从而降低在终产品中的残留风险。方法:建立气相色谱-氢火焰离子化检测器(GC-FID)法,同时检测甲磺酸伏美替尼关键中间体AST 2815中氯甲烷、氯乙烷、乙酰胺3个基因毒性杂质;建立气相色谱-串联质谱(GC-MS/MS)法,检测AST 2815中的4-氯-1-丁醇。结果:氯甲烷、氯乙烷、乙酰胺、4-氯-1-丁醇分别在5.246~104.9、5.228~104.6、2.034~40.68、0.016 05~0.321 1 μg·mL-1浓度范围内,峰面积与质量浓度线性关系良好,r均大于0.999;方法定量限分别为0.016%、0.016%、0.041%、0.000 054%;方法检测限分别为0.005 2%、0.005 2%、0.020%、0.000 018%;平均回收率(n=9)分别为92.8%、95.0%、98.6%、95.1%,RSD (n=9)分别为2.9%、1.4%、3.3%、5.4%;6批样品中4个基因毒性杂质均未检出。结论:方法操作简单,准确可靠,专属性强,灵敏度满足拟定限度,可用于甲磺酸伏美替尼关键中间体AST 2815中4个基因毒性杂质的检测,也可为其他药物中该类基因毒性杂质的痕量分析提供参考。
Objective: To establish the methods for the determination of four genotoxic impurities namely methyl chloride, ethyl chloride, acetamide and 4-chloro-1-butanol in the specification for N2-(2-(dimethylamino) ethyl)-N2-methyl-N5-(4-(1-methyl-1H-indol-3-yl) pyrimidin-2-yl)-6-(2,2,2- trifluoroethoxy) pyridine-2,3,5-triamine (AST 2815), the key intermediate of furmonertinib mesilate, which reduced the residual risk in final product as an in-process control. Methods: The gas chromatography-hydrogen flame ionization detection (GC-FID) method was established for the simultaneous determination of methyl chloride, ethyl chloride and acetamide in the key intermediate AST 2815 of furmonertinib mesilate and gas chromatography-tandem mass spectrometry(GC-MS/MS) method for the determination of 4-chloro-1-butanol in AST 2815. Results: Four impurities indicated good linearity between peak areas and concentration in the range of 5.246-104.9, 5.228-104.6, 2.034-40.68 and 0.016 05-0.321 1 μg·mL-1, respectively, and the r values were all above 0.999. The LOQ of methyl chloride, ethyl chloride, acetamide and 4-chloro-1-butanol was calculated as 0.016%, 0.016%, 0.041%, 0.000 054% and the LOD was 0.005 2%, 0.005 2%, 0.020% and 0.000 018%, respectively. The average recoveries (n=9) were 92.8%, 95.0%, 98.6% and 95.1% and the RSD (n=9) was 2.9%, 1.4%, 3.3% and 5.4%, respectively. The four genotoxic impurities were all not detected in samples from six different batches. Conclusion: The established methods are both simple, rapid, sensitive and accurate, which are applicable for quantification of these four genotoxic impurities in the key intermediate AST 2815. The article can also be referenced for trace analysis of these four genotoxic impurities in other drug substances and drug products.
[1] SHI YK, ZHANG SC, HU XS, et al. Safety, clinical activity, and pharmacokinetics of alflutinib (AST 2818) in patients with advanced NSCLC with EGFR T 790M mutation[J].J Thorac Oncol, 2020, 15(6): 1015
[2] SHI YK, HU XS, ZHANG SC, et al. Efficacy and safety of alflutinib (AST 2818) in patients with T790M mutation-positive NSCLC: a phase Ⅱb multicenter single-arm study[J].J Clin Oncol, 2020, 38(Suppl 15): S9602
[3] LIU XY, LI W, ZHANG YF, et al. Simultaneous determination of alflutinib and its active metabolite in human plasma using liquid chromatography-tandem mass spectrometry[J].J Pharm Biomed Anal, 2019, 176: 112735
[4] 国家药监局. 国家药监局附条件批准甲磺酸伏美替尼片上市[EB/OL].[2021-03-03].https://www.nmpa.gov.cn/yaowen/ypjgyw/20210303144734196.html
National Medical Products Administration. NMPA Granted Conditional Market Approval for Furmonertinib Mesilate Tables[EB/OL].[2021-03-03].https://www.nmpa.gov.cn/yaowen/ypjgyw/20210303144734196.html
[5] 张强, 罗会兵. 吡啶胺基嘧啶衍生物的制备方法及其中间体:中国,201810627166.4[P].2019-12-24
ZHANG Q, LUO HB. Preparation Method of Pyridylaminopyrimidine Derivative and Intermediate Thereof: China, 201810627166.4[P].2019-12-24
[6] ICH. ICH Guideline-Assessment and Control of DNA Reactive (Mutagenic) Impurities in Pharmaceuticals to Limit Potential Carcinogenic Risk M7(R1)[S].2017
[7] SCHULE A, ATES C, PALACIO M, et al. Monitoring and control of genotoxic impurity acetamide in the synthesis of zaurategrast sulfate[J].Org Process Res Dev, 2010, 14(4): 1008
[8] CPDB. All results on each chemical in the carcinogenic potency database(CPDB) [DB/OL].1980 [2021-06-26]. https://files.toxplanet.com/cpdb/chempages/ACETAMIDE.html
[9] 谢含仪, 林云良, 张瑞凌, 等. 基因毒性杂质分析方法和前处理技术的研究进展 [J].药物分析杂志, 2018, 38(10): 1668
XIE HY, LIN YL, ZHANG RL, et al. Advances in analytical methods and pre-treatment techniques for genotoxic impurities[J].Chin J Pharm Anal, 2018, 38(10): 1668
[10] 石富江, 王全军. 基因毒性杂质的毒理学评估策略[J].中国新药杂志, 2021, 30(3): 232
SHI FJ, WANG QJ. Toxicological evaluation strategy of genotoxic impurities[J].Chin J New Drugs, 2021, 30(3): 232
[11] 郑梅, 郑枫, 柳文媛. GC-MS法测定盐酸右美托咪定中氯甲烷和氯乙烷的残留量[J].中南药学, 2018, 16(5): 683
ZHENG M, ZHENG F, LIU WY. Determination of methyl chloride and ethyl chloride in dexmedetomidine hydrochloride by GC-MS[J].Cent South Pharm, 2018, 16(5): 683
[12] 薛泳, 韩洁. 气相色谱电子捕获法测定盐酸氨基葡萄糖中一氯甲烷和一氯乙烷 [J].中国药师, 2018, 21(11): 2082
XUE Y, HAN J. Determination of chloromethane and chloroethane in D-glucosamine hydrochloride by gas chromatography coupled with electron capture detector[J].China Pharm, 2018, 21(11): 2082
[13] 朱峰, 黄静, 王勇军, 等. 气相色谱法测定盐酸贝西沙星中氯乙烷残留量 [J].常州大学学报(自然科学版), 2016, 28(6): 44
ZHU F, HUANG J, WANG YJ, et al. Determination of chloroethane in besifloxacin hydrochloride by gas chromatography[J].J Changzhou Univ(Nat Sci Ed), 2016, 28(6): 44
[14] HARIGAYA K, YAMADA H, YAKU K, et al. Novel sensitive determination method for a genotoxic alkylating agent, 4-chloro-1-butanol, in active pharmaceutical ingredients by LC-ICP-MS employing iodo derivatization[J].Anal Sci, 2014, 30(3): 377
[15] HARIGAYA K, YAMADA H, YAKU K, et al. Development and validation of a sensitive GC-MS method for the determination of alkylating agent, 4-chloro-1-butanol, in active pharmaceutical ingredients[J].Chem Pharm Bull, 2014, 62(4): 395
