title: An electrochemical microfluidic platform for human P450 drug metabolism profiling.
abstract: This paper is the first report of a P450-electrode in a microfluidic format. A 30 μL microfluidic cell was made in poly(methyl methacrylate) containing the inlet, outlet, and reaction chamber with two electrode strips, one of which contains the human cytochrome P450 3A4 covalently bound to gold via a 6-hexanethiol and 7-mercaptoheptanoic acid (1:1) self-assembled monolayer. The electrochemical response of the P450-electrode in the microfluidic cell was tested using four drugs that are known substrates of P450 3A4: quinidine, nifedipine, alosetron and ondansetron. Titration experiments allowed the electrochemical measurements of K(M) for the four drugs, with values of 2.9, 29.1, 113.4, and 114.1 mM, respectively. The K(M) values are found to be in good agreement and correctly ranked with respect to the published literature on human liver microsomes and baculosomes: [ondansetron ≈ alosetron > nifedipine > quinidine]. The results presented in this paper represent a step forward for a rapid evaluation of the interaction of P450 and drug, requiring small volumes of new chemical entities to be tested.
authors.affiliation: Division of Molecular Biosciences, Imperial College London, Biochemistry Building, South Kensington, London, SW7 2AY, United Kingdom.
authors.forename: Andrea
authors.initials: A
authors.lastname: Fantuzzi
authors.forename: Ennio
authors.initials: E
authors.lastname: Capria
authors.forename: Lok Hang
authors.initials: LH
authors.lastname: Mak
authors.forename: Vikash R
authors.initials: VR
authors.lastname: Dodhia
authors.forename: Sheila J
authors.initials: SJ
authors.lastname: Sadeghi
authors.forename: Stephen
authors.initials: S
authors.lastname: Collins
authors.forename: Graham
authors.initials: G
authors.lastname: Somers
authors.forename: Ejaz
authors.initials: E
authors.lastname: Huq
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Gilardi
mesh_terms: D002243:Carbolines / Q000032:analysis; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes; D006801:Humans; D044085:Microfluidics / Q000379:methods*; D008862:Microsomes, Liver / Q000378:metabolism; D009543:Nifedipine / Q000032:analysis; D017294:Ondansetron / Q000032:analysis; D011802:Quinidine / Q000032:analysis; D013379:Substrate Specificity
chemical_list: D002243:Carbolines; C090840:alosetron; D017294:Ondansetron; D003577:Cytochrome P-450 Enzyme System; D009543:Nifedipine; D011802:Quinidine