Inhibition screening method of microsomal UGTs using the cocktail approach

Article Article scientifique Article Inhibition screening method of microsomal UGTs using the cocktail approach eng 2015 author Gradinaru Julieta 767692 author Romand Stéphanie Anne 630412 author Geiser Laurent 87588 author Carrupt Pierre-Alain 135654 author Spaggiari Dany 648593 author Rudaz Serge 0000-0002-4180-5417 28978 35-45 10.1016/j.ejps.2015.02.001 25684194 0928-0987 A rapid method for the simultaneous determination of the in vitro activity of the 10 major human liver UDP-glucuronosyltransferase (UGT) enzymes was developed based on the cocktail approach. Specific substrates were first selected for each UGT: etoposide for UGT1A1, chenodeoxycholic acid for UGT1A3, trifluoperazine for UGT1A4, serotonin for UGT 1A6, isoferulic acid for UGT1A9, codeine for UGT2B4, azidothymidine for UGT2B7, levomedetomidine for UGT2B10, 4-hydroxy-3-methoxymethamphetamine for UGT2B15 and testosterone for UGT2B17. Optimal incubation conditions, including time-based experiments on cocktail metabolism in pooled HLMs that had been performed, were then investigated. A 45- min incubation period was found to be a favorable compromise for all the substrates in the cocktail. Ultra-high pressure liquid chromatography coupled to an electrospray ionization time-of-flight mass spectrometer was used to separate the 10 substrates and their UGT-specific glucuronides in less than 6 min. The ability of the cocktail to highlight the UGT inhibitory potential of xenobiotics was initially proven by using well-known UGT inhibitors (selective and nonselective) and then by relating some of the screening results obtained by using the cocktail approach with morphine glucuronidation (substrate highly glucuronidated by UGT 2B7). All the results were in agreement with both the literature and with the expected effect on morphine glucuronidation. Cocktail approach Phase II metabolism UGT inhibition Human liver microsomes UHPLC–TOF Dewey 615 31 Section des sciences pharmaceutiques 17 false 881 Sciences analytiques European journal of pharmaceutical sciences 71 50314a1c-6341-4a4e-92b1-5b6ba8d3f07b Article (Published version) Restricted 2015(2).pdf 1247486 application/pdf restrictedAccess Faculté des sciences / Section des sciences pharmaceutiques

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Inhibition screening method of microsomal UGTs using the cocktail approach	European journal of pharmaceutical sciences			2015	644	0

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Gradinaru, Julieta