Metabolite profiling of IMID‐2, a novel anticancer molecule of piperazine derivative: In silico prediction, in vitro and in vivo metabolite characterization using UPLC–QTOF–MS/MS

Abstract

IMID‐2, a newly identified piperazine‐based anticancer molecule, has been shown to be cytotoxic against various cancer cell lines. The primary aim of this research was to identify and characterize possible metabolites of the molecule formed during biotransformation. A metabolite identification study was first executed using an in silico tool to predict the possible metabolism sites of IMID‐2. Thereafter, metabolites generated in vitro (rat liver microsomes, rat S9 fractions and human liver microsomes) and in vivo (rat plasma, urine and feces) were identified and characterized employing UPLC–QTOF–MS/MS. A total of eight metabolites, among which were six in phase I and two in phase II reactions, were recognized. The plausible structure of the metabolites and probable metabolic pathway have been established based on the mass fragmentation pattern, mass ppm error, ring double bond calculation and nitrogen rule. The majority of phase I metabolites were generated by N‐oxidation, hydroxylation, oxidative deamination followed by reduction, oxidative dechlorination, N‐dearylation, and N‐dealkylation. Glucuronidation played a significant role in the formation of phase II metabolites of the molecule.

High resolution mass spectrometry‐driven metabolite profiling of baricitinib to report its unknown metabolites and step‐by‐step reaction mechanism of metabolism

“…The supernatant was collected from each sample and subjected to evaporation in a nitrogen evaporator until dry. The dried residue was then reconstituted with the mobile phase and samples were analyzed by U‐HPLC‐Q/TOF and U‐HPLC/LTQ‐Orbitrap‐MS/MS 12–16 …”

Principles and Applications of Ultra-High-Performance Liquid Chromatography

“…It has a vital role in the development of a new chemical entity (NCE). It aids in identifying the active metabolites in so doing, monitoring the possibility of reactive metabolite formations, augmenting pharmacokinetic and pharmacodynamics properties, comparing preclinical with clinical metabolism profile, understanding clearance, and predicting drug-drug or food-drug interactions at early stages of drug discovery [47]. By the time a NCE reaches the development stage, its metabolites' characterization becomes a critical process.…”

“…including in rat liver microsomes (RLM), human liver microsomes (HLM) and rat S9 fraction (RS9), rat plasma, urine, and feces [47,49]. Untargeted metabolomics in diseased plants may offer a fresh viewpoint and advance our comprehension of plant defense mechanisms.…”

Assessment of metabolic stability and pharmacokinetics by LC–MS/MS and establishment of the safe dose of IMID‐2, a novel anticancer molecule under drug discovery

“…Eight metabolites including six phase I and two phase II were identified. Moreover, plausible structures of all the metabolites and the metabolic pathways have already been reported (Panday et al, 2021). In the present study, the molecule was further investigated for its pharmacokinetic behavior, which will help in understanding its complete absorption, distribution, metabolism, excretion (ADME) process.…”