title: Drug Oxidation by Cytochrome P450BM3 : Metabolite Synthesis and Discovering New P450 Reaction Types.
abstract: There is intense interest in late-stage catalytic C-H bond functionalization as an integral part of synthesis. Effective catalysts must have a broad substrate range and tolerate diverse functional groups. Drug molecules provide a good test of these attributes of a catalyst. A library of P450BM3 mutants developed from four base mutants with high activity for hydrocarbon oxidation produced human metabolites of a panel of drugs that included neutral (chlorzoxazone, testosterone), cationic (amitriptyline, lidocaine) and anionic (diclofenac, naproxen) compounds. No single mutant was active for all the tested drugs but multiple variants in the library showed high activity with each compound. The high conversions enabled full product characterization that led to the discovery of the new P450 reaction type of oxidative decarboxylation of an α-hydroxy carboxylic acid and the formation a protected imine from an amine, offering a novel route to α-functionalization of amines. The substrate range and varied product profiles suggest that this library of enzymes is a good basis for developing late-stage C-H activation catalysts.
authors: Department of Chemistry, University of Oxford, Inorganic Chemistry Laboratory, South Parks Rd., Oxford OX1 3QR (UK).
authors: Xinkun
authors: X
authors: Ren
authors: Department of Chemistry, University of Oxford, Inorganic Chemistry Laboratory, South Parks Rd., Oxford OX1 3QR (UK).
authors: Jake A
authors: JA
authors: Yorke
authors: Department of Chemistry, University of Oxford, Inorganic Chemistry Laboratory, South Parks Rd., Oxford OX1 3QR (UK).
authors: Emily
authors: E
authors: Taylor
authors: College of Life Sciences and The State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin 300071 (P. R. China).
authors: Ting
authors: T
authors: Zhang
authors: College of Life Sciences and The State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin 300071 (P. R. China). zhouwh@nankai.edu.cn.
authors: Weihong
authors: W
authors: Zhou
authors: Department of Chemistry, University of Oxford, Inorganic Chemistry Laboratory, South Parks Rd., Oxford OX1 3QR (UK). luet.wong@chem.ox.ac.uk.
authors: Luet Lok
authors: LL
authors: Wong
mesh_terms: D002384:Catalysis; D002753:Chlorzoxazone / Q000737:chemistry* / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000378:metabolism*; D004008:Diclofenac / Q000737:chemistry*; D006801:Humans; D006860:Hydrogen Bonding; D007700:Kinetics; D009288:Naproxen / Q000737:chemistry*; D010084:Oxidation-Reduction; D015202:Protein Engineering; D013739:Testosterone / Q000737:chemistry* / Q000378:metabolism
chemical_list: D004008:Diclofenac; D013739:Testosterone; D009288:Naproxen; D003577:Cytochrome P-450 Enzyme System; D002753:Chlorzoxazone
keywords: CH activation; cytochrome P450; decarboxylation; drug metabolism; protein engineering