Abstract The fragmentation pattern of certain isatin-based compounds was carried out using collision-induced dissociation inside the triple quadrupole mass analyzer. These data were used as a clue for the identification of metabolites of the recently reported isatin-based antiproliferative agent, namely, N′-[5-bromo-1-methyl-2-oxo-1,2-dihydro-3H-indol-3-ylidene]-5-methoxy-1H-indole-2-carbohydrazide (1) in rat liver microsomes (RLMs) using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Prediction of the vulnerable sites for metabolic pathways in compound 1 was performed by WhichP450 module of StarDrop software. In vitro metabolites for compound 1 were identified with the aid of rat liver microsomes. The in silico data were utilized as a guide for the practical work. Compound 1 was metabolized into three (hydroxylated, reduced and O-demethylated) metabolites in RLMs in the presence of NADPH. The chemical structures of those metabolites were elucidated, and the metabolic pathways were proposed by comparing the fragmentation pattern of the isatin–indole conjugates 1–7. The data presented in this paper provided useful information on the effect of different substituents on the ionization/fragmentation processes and can be used in the characterization of isatin derivatives. In silico toxicity assessments for the title compounds 1–7 and for the metabolites of compound 1 were conducted utilizing the deductive estimation of risk from existing knowledge (DEREK) module of StarDrop software.

中文翻译：

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某些靛红-吲哚抗增殖偶联物的碎片模式，用于通过液相色谱串联质谱法鉴定其在大鼠肝微粒体中的体外代谢谱

摘要 在三重四极杆质量分析器内使用碰撞诱导解离对某些靛红基化合物的裂解模式进行了分析。这些数据被用作鉴定最近报道的基于靛红的抗增殖剂的代谢物的线索，即 N'-[5-bromo-1-methyl-2-oxo-1,2-dihydro-3H-indol-使用液相色谱-串联质谱 (LC-MS/MS) 分析大鼠肝微粒体 (RLM) 中的 3-亚基]-5-甲氧基-1H-吲哚-2-碳酰肼 (1)。StarDrop 软件的WhichP450 模块对化合物1 中代谢途径的脆弱位点进行了预测。在大鼠肝微粒体的帮助下鉴定了化合物 1 的体外代谢物。计算机数据被用作实际工作的指南。化合物 1 被代谢为三种（羟基化、在 NADPH 存在下，RLM 中还原和 O-去甲基化的）代谢物。阐明了这些代谢物的化学结构，并通过比较靛红-吲哚缀合物 1-7 的碎片模式提出了代谢途径。本文中提供的数据提供了有关不同取代基对电离/断裂过程的影响的有用信息，可用于表征靛红衍生物。利用 StarDrop 软件的现有知识 (DEREK) 模块的风险推断性估计对标题化合物 1-7 和化合物 1 的代谢物进行了计算机毒性评估。代谢途径是通过比较靛红-吲哚缀合物 1-7 的碎片模式提出的。本文中提供的数据提供了有关不同取代基对电离/断裂过程影响的有用信息，可用于表征靛红衍生物。利用 StarDrop 软件的现有知识 (DEREK) 模块的风险推断性估计对标题化合物 1-7 和化合物 1 的代谢物进行了计算机毒性评估。代谢途径是通过比较靛红-吲哚缀合物 1-7 的碎片模式提出的。本文中提供的数据提供了有关不同取代基对电离/断裂过程的影响的有用信息，可用于表征靛红衍生物。利用 StarDrop 软件的现有知识 (DEREK) 模块的风险推断性估计对标题化合物 1-7 和化合物 1 的代谢物进行了计算机毒性评估。