Electrochemical simulation of metabolism for antitumor-active imidazoacridinone C-1311 and in silico prediction of drug metabolic reactions

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 169; pp. 269 - 278
• Main Authors: Potęga, Agnieszka, Żelaszczyk, Dorota, Mazerska, Zofia
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 30-05-2019
• Subjects: Aminoacridines - metabolism; Antineoplastic Agents - metabolism; Antitumor compound; Biochemical Phenomena - physiology; Computer Simulation; Cytochrome P-450 Enzyme System - metabolism; Cytochrome P450; Electrochemical Techniques - methods; Electrochemistry - methods; Electrodes; Humans; In silico site of metabolism prediction; In vitro drug metabolism; Inactivation, Metabolic - physiology; Metabolite electrosynthesis; Microsomes, Liver - metabolism; On-line electrochemistry-mass spectrometry; Oxidation-Reduction; Spectrometry, Mass, Electrospray Ionization - methods; Tandem Mass Spectrometry - methods; On-line electrochemistry-mass spectrometry; Q-TOF; MS; Cytochrome P450; In silico site of metabolism prediction; Antitumor compound; C-1311; MW; In vitro drug metabolism; MS/MS; Metabolite electrosynthesis; LC; m/z; ESI; P450; FA; EC
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/j.jpba.2019.03.017

•Phase I metabolism of C-1311 was successfully simulated in an electrochemical cell.•The products of N-dealkylation and aliphatic hydroxylation reactions were detected.•In silico analysis was used for the prediction of P450-mediated metabolites.•We observed a good accordance between electrochemical and in silico results.•Electrochemical and in silico methods are fast alternatives for enzymatic assays.
The metabolism of antitumor-active 5-diethylaminoethylamino-8-hydroxyimidazoacridinone (C-1311) has been investigated widely over the last decade but some aspects of molecular mechanisms of its metabolic transformation are still not explained. In the current work, we have reported a direct and rapid analytical tool for better prediction of C-1311 metabolism which is based on electrochemistry (EC) coupled on-line with electrospray ionization mass spectrometry (ESI-MS). Simulation of the oxidative phase I metabolism of the compound was achieved in a simple electrochemical thin-layer cell consisting of three electrodes (ROXY™, Antec Leyden, the Netherlands). We demonstrated that the formation of the products of N-dealkylation reactions can be easily simulated using purely instrumental approach. Newly reported products of oxidative transformations like hydroxylated or oxygenated derivatives become accessible. Structures of the electrochemically generated metabolites were elucidated on the basis of accurate mass ion data and tandem mass spectrometry experiments. In silico prediction of main sites of C-1311 metabolism was performed using MetaSite software. The compound was evaluated for cytochrome P450 1A2-, 3A4-, and 2D6-mediated reactions. The results obtained by EC were also compared and correlated with those of reported earlier for conventional in vitro enzymatic studies in the presence of liver microsomes and in the model peroxidase system. The in vitro experimental approach and the in silico metabolism findings showed a quite good agreement with the data from EC/ESI-MS analysis. Thus, we conclude here that the electrochemical technique provides the promising platform for the simple evaluation of drug metabolism and the reaction mechanism studies, giving first clues to the metabolic transformation of pharmaceuticals in the human body.