Ropinirole HCl, β‐nicotinamide adenine dinucleotide 2′‐phosphate (NADPH) and fluvoxamine maleate were purchased from Sigma‐Aldrich (St. Louis, MO, USA). Despropyl ropinirole HCl and 7‐hydroxy ropinirole HBr were purchased as reference standards from TLC Pharmaceutical Standards LTD (Aurora, ON, Canada). All solvents used were of analytical grade. The stock solutions of ropinirole and fluvoxamine were prepared in purified (MilliQ) water and the metabolite standards in methanol.
Human and dog hepatocytes, as well as Krebs–Henseleit buffer (KHB), were purchased from BioIVT (Westbury, NY, USA). The dog hepatocytes were a pool of four male beagles, whereas human hepatocytes were a mixed gender pool of 50 donors. Canine recombinant CYP (rCYP) Bactosomes (co‐expressed with dog CYP reductase in Escherichia coli) were obtained from Cypex LTD (Dundee, Scotland). Ropinirole was tested in all available canine CYP isoforms (CYP1A1, CYP1A2, CYP1B1, CYP2C11, CYP2C21, CYP2C41, CYP2D15, CYP3A12 and CYP3A26).
Human and dog hepatocytes, as well as Krebs–Henseleit buffer (KHB), were purchased from BioIVT (Westbury, NY, USA). The dog hepatocytes were a pool of four male beagles, whereas human hepatocytes were a mixed gender pool of 50 donors. Canine recombinant CYP (rCYP) Bactosomes (co‐expressed with dog CYP reductase in Escherichia coli) were obtained from Cypex LTD (Dundee, Scotland). Ropinirole was tested in all available canine CYP isoforms (CYP1A1, CYP1A2, CYP1B1, CYP2C11, CYP2C21, CYP2C41, CYP2D15, CYP3A12 and CYP3A26).