Identification of metabolic pathways involved in the biotransformation of eslicarbazepine acetate using UPLC-MS/MS, human microsomal enzymes and in silico studies

In the last few decades, metabolite identification and detection have become an integral component of drug discovery process. The detection and depiction of an identified metabolite is significant for the development of safe and effective therapeutic agents as these metabolites may be pharmacologically active or toxic in nature. A (UPLC-MS/MS) method was developed for the quantification of eslicarbazepine acetate metabolite (M3), cis-10,11-dihydro-5H-dibenz[b,f]azepine-10,11-diol, in rat plasma. Eslicarbazepine acetate was extracted from rat plasma by precipitation technique using acetonitrile as a precipitating solvent. Chromatographic separation was accomplished using acetonitrile-0.01 M potassium dihydrogen phosphate (60:40, v/v) as mobile phase at a flow rate of 0.2 ml/min on Waters Acquity BEH 150 x 2.1 mm, 1.7 mm, C18 column at 30'C. The linearity of the calibration curve was 5-100 ng/ml for metabolite M3. The overall intra-and inter-day precision did not exceed 3%. In silico, in vitro and in vivo approaches have been used to facilitate the detection and depiction of eslicarbazepine acetate metabolites. MetabolExpert was used to predict the probable metabolites of the compounds. Metabolite of eslicarbazepine acetate was identified using human liver microsomes.

Automated summary

Metabolite identification and detection have become crucial in drug discovery in the last few decades. A method using UPLC-MS/MS was developed to quantify a specific metabolite in rat plasma, and the metabolite of eslicarbazepine acetate was successfully identified. Various approaches were used to aid in the detection and depiction of metabolites in this study.