title: Discovery of novel PDE10 inhibitors by a robust homogeneous screening assay.
abstract: OBJECTIVE
  To develop a homogeneous assay for high-throughput screening (HTS) of inhibitors of phosphodiesterase 10 (PDE10).
  
  
  METHODS
  Purified human PDE10 enzyme derived from E coli, [(3)H]-cAMP and yttrium silicate microbeads were used to develop an HTS assay based on the scintillation proximity assay (SPA) technology. This method was applied to a large-scale screening campaign against a diverse compound library and subsequent confirmation studies. Preliminary structure-activity relationship (SAR) studies were initiated through limited structural modifications of the hits.
  
  
  RESULTS
  The IC50 value of the control compound (papaverine) assessed with the SPA approach was comparable and consistent with that reported in the literature. Signal to background (S/B) ratio and Z' factor of the assay system were evaluated to be 5.24 and 0.71, respectively. In an HTS campaign of 71 360 synthetic and natural compounds, 67 hits displayed reproducible PDE10 inhibition, of which, 8 were chosen as the scaffold for structural modifications and subsequent SAR analysis.
  
  
  CONCLUSIONS
  The homogeneous PDE10 SPA assay is an efficient and robust tool to screen potential PDE10 inhibitors. Preliminary SAR studies suggest that potent PDE10 inhibitors could be identified and developed through this strategy.
authors.affiliation: The National Center for Drug Screening and the State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Qun-yi
authors.initials: QY
authors.lastname: Li
authors.forename: Ming-kai
authors.initials: MK
authors.lastname: Xu
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
authors.forename: Claus Tornby
authors.initials: CT
authors.lastname: Christoffersen
authors.forename: Ming-wei
authors.initials: MW
authors.lastname: Wang
mesh_terms: D055808:Drug Discovery / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D010208:Papaverine / Q000737:chemistry / Q000494:pharmacology*; D010726:Phosphodiesterase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D010726:Phosphodiesterase Inhibitors; D010208:Papaverine; C119843:PDE10A protein, human; D010727:Phosphoric Diester Hydrolases