抗HIV先导物DAAN-10341的体外代谢产物筛查

Metabolite Profiling of Anti-HIV Lead Compound DAAN-10341 in vitro

目的应用超高效液相色谱-四极杆飞行时间质谱联用技术(UPLC-QTOF-MS/MS)并辅以代谢数据采集和处理软件,对新型非核苷类逆转录酶抑制剂(NNRTIs)的先导物DAAN-10341在大鼠肝微粒体的代谢产物进行快速筛查和鉴定,并对其结构中的代谢位点进行分析。方法应用质量亏损过滤技术在UPLC-QTOF-MS/MS上比较分析0 h与2 h孵育样品的数据,在大鼠肝微粒体孵育液中筛查得到了DAAN-10341的16个代谢产物。应用仪器的碰撞能量梯度功能(MSE)对部分产物进行MS2分析,获得代谢产物的碎片,推断碎裂途径及产物结构。结果先导物DAAN-10341在大鼠肝微粒体中的Ⅰ相代谢反应以氧化(羟基化)为主;Ⅱ相代谢主要以葡糖醛酸结合反应与谷胱甘肽结合反应为主。结论本文通过分析新型抗HIV先导物DAAN-10341的结构和代谢转化,发现易于发生代谢的位点和结构特征,为该类先导物结构优化和后续的临床前评价提供科学指导。

Objective The metabolite profiling of DAAN-10341, the lead compound from new classes of non-nucleoside reverse transcriptase inhibitors （NNRTIs） ,was performed using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry （LC-QTOF-MS/MS）, with the assistance of a metabolite data processing software. Methods Using the mass defect filter （MDF） technique, the data acquired from the 0 h-incubate and the 2 h-incubate were compared and analyzed with the software. A total of 16 metabolites were discovered for DAAN-10341 in the 2 h-incubate. The MS2 spectra for some metabolites were obtained using the MSE technique to get fragment ions for structural elucidation. Results Major metabolic pathways were identified. The compound could undergo extensive metabolism in rat liver rnicrosomes. The major phase Ⅰ reaction was oxidation/hydroxylation. The major reactions of phase Ⅱ were glucuronide conjugation and S-glutathione conjugation. Conclusion The structure-metabolic pathway relationship was analyzed to reveal metabolic soft spots and the structure characteristics of the compound, which may facilitate structural optimization of lead compounds,as well as preclinical evaluation of new chemical entities.