Integration of virtual screening with high-throughput flow cytometry to identify novel small molecule formylpeptide receptor antagonists. Academic Article uri icon

start page

  • 1301

end page

  • 1310

abstract

  • The formylpeptide receptor (FPR) family of G-protein-coupled receptors contributes to the localization and activation of tissue-damaging leukocytes at sites of chronic inflammation. We developed a FPR homology model and pharmacophore (based on the bovine rhodopsin crystal structure and known FPR ligands, respectively) for in silico screening of approximately 480,000 drug-like small molecules. A subset of 4324 compounds that matched the pharmacophore was then physically screened with the HyperCyt flow cytometry platform in high-throughput, no-wash assays that directly measure human FPR binding, with samples (each approximately 2500 cells in 2 microl) analyzed at 40/min. From 52 confirmed hits (1.2% hit rate), we identified 30 potential lead compounds (inhibition constant, Ki= 1-32 microM) representing nine distinct chemical families. Four compounds in one family were weak partial agonists. All others were antagonists. This virtual screening approach improved the physical screening hit rate by 12-fold (versus 0.1% hit-rate in a random compound collection), providing an efficient process for identifying small molecule antagonists.

date/time value

  • 2005

Digital Object Identifier (DOI)

  • 10.1124/mol.105.014068

PubMed Identifier

  • 16118363

volume

  • 68

number

  • 5

keywords

  • Amino Acid Sequence
  • Animals
  • Cattle
  • Drug Evaluation, Preclinical
  • Flow Cytometry
  • Models, Molecular
  • Molecular Sequence Data
  • Receptors, Formyl Peptide