title: Cell-based multi-substrate assay coupled to UHPLC-ESI-MS/MS for a quick identification of class-specific HDAC inhibitors.
abstract: Histone deacetylases (HDAC) are involved in several diseases including cancer, cardiovascular and neurodegenerative disorders, and the search for inhibitors is a current topic in drug discovery. Four HDAC inhibitors have already been approved by the FDA for cancer therapy and others are under clinical studies. However, the clinical utility of some of them is limited because of unfavorable toxicities associated with their broad range of HDAC inhibitory effects. Toxicity could be decreased by using HDAC inhibitors with improved specificity. To date, the most popular screening assays are based on fluorescence-labeled substrates incubated with an enzymatic source (cells extracts or recombinant isoforms). Here, we describe a high-throughput cell-based UHPLC-ESI-MS/MS assay able to rapidly predict activity against HDAC1 and HDAC6 in a cell environment. This method is predicted to be a useful tool to accelerate the search for class-selective HDAC inhibitors in drug discovery.
authors.affiliation: a School of Pharmaceutical Sciences, University of Geneva, University of Lausanne , Geneva , Switzerland.
authors.forename: Vincent
authors.initials: V
authors.lastname: Zwick
authors.affiliation: a School of Pharmaceutical Sciences, University of Geneva, University of Lausanne , Geneva , Switzerland.
authors.forename: Claudia
authors.initials: C
authors.lastname: Simões-Pires
authors.affiliation: a School of Pharmaceutical Sciences, University of Geneva, University of Lausanne , Geneva , Switzerland.
authors.forename: Muriel
authors.initials: M
authors.lastname: Cuendet
mesh_terms: D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006367:HeLa Cells; D056284:Histone Deacetylase 1 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000073864:Histone Deacetylase 6; D056572:Histone Deacetylase Inhibitors / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology*; D006655:Histone Deacetylases / Q000378:metabolism*; D006801:Humans; D008968:Molecular Conformation; D021241:Spectrometry, Mass, Electrospray Ionization; D013329:Structure-Activity Relationship
chemical_list: D056572:Histone Deacetylase Inhibitors; C110550:HDAC1 protein, human; C433811:HDAC6 protein, human; D056284:Histone Deacetylase 1; D000073864:Histone Deacetylase 6; D006655:Histone Deacetylases
keywords: Class I HDAC; HDAC6; high-throughput screening; histone deacetylase; isoform selectivity