阿伐那非潜在遗传毒性杂质E的合成及致突变性风险研究

Synthesis and mutagenicity risk of Avanafil potential genotoxic impurity E

关键词:  阿伐那非,酰肼杂质,Ames试验,遗传毒性,定量构效关系

Key words:  Avanafil, hydrazide  impurity, Ames  test, genotoxicity, QSAR

摘要:

目的：对阿伐那非合成工艺中具有警示结构的潜在有关物质进行致突变性评价研究，为阿伐那非有关物质的分类分级提供指导和依据。方法：通过化学方法定向合成阿伐那非含警示结构的潜在遗传毒性杂质E，按照ICH M7指导原则对其进行评价和分类。分别采用基于专家规则和统计学原理的2种互补的定量构效关系(QSAR)评价系统(Derek和Sarah)对杂质E的遗传毒性进行初步预测，并展开Ames试验对杂质E进一步验证。结果：杂质E的Derek预测结果为阳性，依据 ICH M7 指导原则的分类为第 3 类。在Ames验证试验中，受试物在62.5～1000 μg/皿剂量范围内，无论S9存在与否的情况下，回复突变菌落数均未超过溶剂对照组菌落数的2倍且不呈剂量-反应关系，试验结果显阴性。结论：杂质E的致突变性为阴性，推翻了基于警示结构的疑虑，依据ICH M7中的第5类即非突变性杂质进行控制。

Abstract:

OBJECTIVE: To evaluate the mutagenicity of potential process impurities with warning structure in the synthetic process of avanafil, so as to provide guidance and basis for the classification and control of related substances of avanafil. METHODS: The potentially genotoxic impurity E (Imp-E) of Avanafil with warning hydrazide-structure was synthesized by chemical method and preliminary predictions were made according to ICH M7 guidelines. Ames test was then used to further verify the Imp-E. RESULTS: Derek prediction results for Imp-E were all positive, and were classified into Class 3 according to the ICH M7 guidelines. In Ames test, the number of revertant colonies of Imp-E in the presence or absence of metabolic activation system S9 in the range of 62.5～1000 μg/per plate of test substance was less than 2 times of that in the solvent control group, and the test result was negative. CONCLUSION: The bacterial mutation test results of Imp-E are all negative, which can overturn the doubts based on the structure, and these impurities can be controlled as Class 5 in ICH M7, which namely non mutagenic impurity.