title: NMR-based platform for fragment-based lead discovery used in screening BRD4-targeted compounds.
abstract: OBJECTIVE
  Fragment-based lead discovery (FBLD) is a complementary approach in drug research and development. In this study, we established an NMR-based FBLD platform that was used to screen novel scaffolds targeting human bromodomain of BRD4, and investigated the binding interactions between hit compounds and the target protein.
  
  
  METHODS
  1D NMR techniques were primarily used to generate the fragment library and to screen compounds. The inhibitory activity of hits on the first bromodomain of BRD4 [BRD4(I)] was examined using fluorescence anisotropy binding assay. 2D NMR and X-ray crystallography were applied to characterize the binding interactions between hit compounds and the target protein.
  
  
  RESULTS
  An NMR-based fragment library containing 539 compounds was established, which were clustered into 56 groups (8-10 compounds in each group). Eight hits with new scaffolds were found to inhibit BRD4(I). Four out of the 8 hits (compounds 1, 2, 8 and 9) had IC50 values of 100-260 μmol/L, demonstrating their potential for further BRD4-targeted hit-to-lead optimization. Analysis of the binding interactions revealed that compounds 1 and 2 shared a common quinazolin core structure and bound to BRD4(I) in a non-acetylated lysine mimetic mode.
  
  
  CONCLUSIONS
  An NMR-based platform for FBLD was established and used in discovery of BRD4-targeted compounds. Four potential hit-to-lead optimization candidates have been found, two of them bound to BRD4(I) in a non-acetylated lysine mimetic mode, being selective BRD4(I) inhibitors.
authors.affiliation: Department of Analytical Chemistry, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; China & University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Jun-Lan
authors.initials: JL
authors.lastname: Yu
authors.affiliation: CAS Key Laboratory of Receptor Research, Drug Discovery and Design Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Tian-Tian
authors.initials: TT
authors.lastname: Chen
authors.affiliation: Department of Analytical Chemistry, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; China & University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Zhou
authors.affiliation: Department of Analytical Chemistry, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; China & University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Fu-Lin
authors.initials: FL
authors.lastname: Lian
authors.affiliation: Department of Analytical Chemistry, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; China & University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Xu-Long
authors.initials: XL
authors.lastname: Tang
authors.affiliation: Department of Analytical Chemistry, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; China & University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wen
authors.affiliation: State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Jing-Kang
authors.initials: JK
authors.lastname: Shen
authors.affiliation: CAS Key Laboratory of Receptor Research, Drug Discovery and Design Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Ye-Chun
authors.initials: YC
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
authors.forename: Bing
authors.initials: B
authors.lastname: Xiong
authors.affiliation: Department of Analytical Chemistry, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; China & University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Nai-Xia
authors.initials: NX
authors.lastname: Zhang
mesh_terms: D018797:Cell Cycle Proteins; D055808:Drug Discovery / Q000379:methods*; D005454:Fluorescence Polarization; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors*; D011485:Protein Binding; D054852:Small Molecule Libraries; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000037:antagonists & inhibitors*; D000092468:Bromodomain Containing Proteins
chemical_list: D018797:Cell Cycle Proteins; D009687:Nuclear Proteins; D054852:Small Molecule Libraries; D014157:Transcription Factors; C441289:BRD4 protein, human; D000092468:Bromodomain Containing Proteins