DFT方法辅助的DNA碱基加合物高通量筛选技术在杂质基因毒性评估确证中的应用基础研究

Drug genotoxic impurities (GTIs) can cause DNA damage directly or indirectly, and then result in the mutagenic or carcinogenic effects. Even trace GTIs can cause significant risk for patient’s health, so genotoxic evaluation of drug impurities is a hot topic in recent years. High screening accuracy and throughput are the research objectives, so a method to directly exlpore DNA base adducts which is foundmental genotoxic scaffold using aromatic amines as model would be developed in this research. First, possible DNA base adduct species and corresponding toxicity will be predicted by Density Functional Theory (DFT), and then qualitative identification and quantitative characterization of adducts will be executed with high resolution liquid chromatography-mass technology. With the data of possible DNA base adducts species, amounts, dose- and time-effect in hand, then we can draw a more accurate conclusion about genotoxicity, which is important for fully understanding the genetic toxicity of impurities. This study would provide a guideline to establish safe window of a new drug, and lay a foundation for setting up impurity-specific quality control standard.

药物基因毒性杂质（GTIs）可直接或间接损伤细胞DNA，从而产生致突变或致癌作用，极微量的GTIs即可对人体产生重大安全风险，因此药物杂质的基因毒性评估是当前研究热点。目前用于基因毒性筛选和验证的实验方法在结果准确性方面有待提高，本研究拟以芳香胺类杂质为例，基于反映化合物基因毒性的最小因子，创新性地提出了以DNA碱基加合物为判定指标的高分辨液相色谱-质谱的基因毒性评估确证策略。在密度泛函理论（DFT）计算方法预测结果的基础上，聚焦于生物体内形成的DNA碱基加合物的定性鉴定和定量表征，着力解决筛选的高通量和评估结果的高准确度问题。初步明确生物体内形成的DNA加合物种类和含量，研究不同加合物的量效、时效关系，据此评价化合物的基因毒性。该研究对于全面正确理解杂质的基因毒性具有重要意义，为针对具体杂质建立特异性质控标准提供技术支持，为准确评估GTIs的暴露和健康风险奠定基础。