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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: To determine the effect of novel non-nucleoside reverse transcriptase inhibitors (NNRTIs) using ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS / MS) combined with metabolic data acquisition and processing software The leader DAAN-10341 was rapidly screened and identified in the metabolites of rat liver microsomes and the metabolic loci in its structure were analyzed. Methods The data of 0 h and 2 h incubated samples were compared by UPLC-QTOF-MS / MS with mass loss filter technique. Sixteen metabolites of DAAN-10341 were screened in rat liver microsomal incubation solution. The collision energy gradient function (MSE) of the instrument was used to perform MS2 analysis on some products to obtain the fragments of the metabolites and deduce the fragmentation pathway and product structure. Results The phase Ⅰ metabolic reaction of DAAN-10341 in rat liver microsomes was mainly oxidative (hydroxylation). The phase Ⅱ metabolism mainly consisted of glucuronic acid binding reaction and glutathione binding reaction. Conclusion The structure and metabolic transformation of DAAN-10341, a novel anti-HIV leader, were found in this paper to find sites and structural features that are prone to be metabolized and to provide scientific guidance for structural optimization and subsequent preclinical evaluation of such precursors.