title: Cell-based assay for screening 11beta-hydroxysteroid dehydrogenase inhibitors using liquid chromatography/tandem mass spectrometry detection.
abstract: Cortisol is an important glucocorticoid that regulates many physiological pathways by activating various intracellular receptors. The type 1 isozyme of 11beta-hydroxysteroid dehydrogenase (11beta-HSD1) functions in vivo predominantly as a reductase by converting cortisone into cortisol. A high-throughput liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed to screen for inhibitors of 11beta-HSD1 by monitoring cortisol and cortisone simultaneously. The injection cycle time can be as fast as 1 min/sample, making it amenable to the analysis of large numbers of the cell-assay samples in the screening of 11beta-HSD inhibitors. The reductase and dehydrogenase activities of 11beta-HSD1 are assessed separately.
authors.affiliation: Takeda San Diego, Inc., 10410 Science Center Dr., San Diego, CA 92121, USA. ron.xu@takedasd.com
authors.forename: Rongda
authors.initials: R
authors.lastname: Xu
authors.forename: Bi-Ching
authors.initials: BC
authors.lastname: Sang
authors.forename: Marc
authors.initials: M
authors.lastname: Navre
authors.forename: Daniel B
authors.initials: DB
authors.lastname: Kassel
mesh_terms: D042842:11-beta-Hydroxysteroid Dehydrogenases / Q000037:antagonists & inhibitors* / Q000235:genetics; D000818:Animals; D002138:Calibration; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003348:Cortisone / Q000032:analysis*; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D006854:Hydrocortisone / Q000032:analysis*; D051379:Mice; D051381:Rats; D053719:Tandem Mass Spectrometry
chemical_list: D004791:Enzyme Inhibitors; D042842:11-beta-Hydroxysteroid Dehydrogenases; D003348:Cortisone; D006854:Hydrocortisone