title: Novel KCNQ2 channel activators discovered using fluorescence-based and automated patch-clamp-based high-throughput screening techniques.
abstract: OBJECTIVE
  To establish an improved, high-throughput screening techniques for identifying novel KCNQ2 channel activators.
  
  
  METHODS
  KCNQ2 channels were stably expressed in CHO cells (KCNQ2 cells). Thallium flux assay was used for primary screening, and 384-well automated patch-clamp IonWorks Barracuda was used for hit validation. Two validated activators were characterized using a conventional patch-clamp recording technique.
  
  
  RESULTS
  From a collection of 80 000 compounds, the primary screening revealed a total of 565 compounds that potentiated the fluorescence signals in thallium flux assay by more than 150%. When the 565 hits were examined in IonWorks Barracuda, 38 compounds significantly enhanced the outward currents recorded in KCNQ2 cells, and were confirmed as KCNQ2 activators. In the conventional patch-clamp recordings, two validated activators ZG1732 and ZG2083 enhanced KCNQ2 currents with EC50 values of 1.04±0.18 μmol/L and 1.37±0.06 μmol/L, respectively.
  
  
  CONCLUSIONS
  The combination of thallium flux assay and IonWorks Barracuda assay is an efficient high-throughput screening (HTS) route for discovering KCNQ2 activators.
authors.affiliation: CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, State Key Laboratory of Drug Research, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Jin-feng
authors.initials: JF
authors.lastname: Yue
authors.affiliation: CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, State Key Laboratory of Drug Research, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Guan-hua
authors.initials: GH
authors.lastname: Qiao
authors.affiliation: The National Center for Drug Screening, Shanghai Institute of Materia Medica, State Key Laboratory of Drug Research, Chinese Academy of Sciences, Shanghai 201203, China.|Department of Chemistry and Institute of Medicinal Chemistry, East China Normal University, Shanghai 200062, China.
authors.forename: Ni
authors.initials: N
authors.lastname: Liu
authors.affiliation: The National Center for Drug Screening, Shanghai Institute of Materia Medica, State Key Laboratory of Drug Research, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Fa-jun
authors.initials: FJ
authors.lastname: Nan
authors.affiliation: CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, State Key Laboratory of Drug Research, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Zhao-bing
authors.initials: ZB
authors.lastname: Gao
mesh_terms: D000818:Animals; D016466:CHO Cells; D003412:Cricetulus; D062126:Databases, Chemical; D005453:Fluorescence; D057166:High-Throughput Screening Assays; D015640:Ion Channel Gating; D051658:KCNQ2 Potassium Channel / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D013793:Thallium / Q000378:metabolism
chemical_list: D051658:KCNQ2 Potassium Channel; D013793:Thallium