Fragmentation pattern of certain isatin-indole antiproliferative conjugates with application to identify their in vitro metabolic profiles in rat liver microsomes by liquid chromatography tandem mass spectrometry

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-carbohyclrazide (1) in rat liver micro-somes (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 0-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.