title: Biosynthesis of 6beta-hydroxymethyltestosterone using bovine hepatocyte cultures.
abstract: Usually performed to investigate biotransformations of xenobiotics, in vitro liver models could become useful tools for the synthesis of not commercially available compounds. In this study, bovine hepatocyte cultures were used to biosynthesise, on the laboratory scale, one major metabolite of methyltestosterone: 6beta-hydroxymethyltestosterone. After incubation of bovine hepatocytes with methyltestosterone for 24 h, culture medium was removed and stored at -20 degrees C until analysis. The sample was extracted and purified on a reversed-phase HPLC system. The metabolite of interest was then analysed in LC-MS and GC-MS for structural identification. The purity and the isomery of the 6 and 17 positions were confirmed by NMR analyses. This first success in producing purified 6beta-hydroxymethyltestosterone from bovine hepatocyte cultures allowed us to consider that in vitro liver models could be reliable tools for standard biosynthesis.
authors.affiliation: LDH-LNR, Ecole Nationale Vétérinaire (Ministère de l'Agriculture), BP 50707, F-44307, Cedex 3, Nantes, France.
authors.forename: A
authors.initials: A
authors.lastname: Clouet-Dumas
authors.forename: B
authors.initials: B
authors.lastname: Le Bizec
authors.forename: M
authors.initials: M
authors.lastname: Le Pape
authors.forename: D
authors.initials: D
authors.lastname: Maume
authors.forename: F
authors.initials: F
authors.lastname: Monteau
authors.forename: F
authors.initials: F
authors.lastname: Andre
mesh_terms: D000818:Animals; D001711:Biotransformation; D002417:Cattle; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D022781:Hepatocytes / Q000378:metabolism*; D007536:Isomerism; D009682:Magnetic Resonance Spectroscopy; D008777:Methyltestosterone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D013739:Testosterone / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism*; D015262:Xenobiotics / Q000378:metabolism
chemical_list: C416844:6-hydroxymethyltestosterone; D017077:Culture Media, Conditioned; D015262:Xenobiotics; D013739:Testosterone; D008777:Methyltestosterone