An Electrochemical Microfluidic Platform for Human P450 Drug Metabolism Profiling

摘要

This paper is the first report of a P450-electrode in anmicrofluidic format. A 30 μL microfluidic cell was madenin poly(methyl methacrylate) containing the inlet, outlet,nand reaction chamber with two electrode strips, one ofnwhich contains the human cytochrome P450 3A4 covalentlynbound to gold via a 6-hexanethiol and 7-mercaptoheptanoicnacid (1:1) self-assembled monolayer. Thenelectrochemical response of the P450-electrode in thenmicrofluidic cell was tested using four drugs that arenknown substrates of P450 3A4: quinidine, nifedipine,nalosetron and ondansetron. Titration experiments allowednthe electrochemical measurements of KM for the fourndrugs, with values of 2.9, 29.1, 113.4, and 114.1nmM, respectively. The KM values are found to be inngood agreement and correctly ranked with respect tonthe published literature on human liver microsomesnand baculosomes: [ondansetron ≈ alosetron > nifedipinen> quinidine]. The results presented in thisnpaper represent a step forward for a rapid evaluationnof the interaction of P450 and drug, requiring smallnvolumes of new chemical entities to be tested.