A cell-based fluorescent assay for FAP inhibitor discovery

Bioorg Med Chem Lett. 2020 Jul 15;30(14):127253. doi: 10.1016/j.bmcl.2020.127253. Epub 2020 May 11.

Abstract

To facilitate the discovery of FAP inhibitors, a convenient cell-based fluorescent assay was developed by using a commonly available U87MG cell line and a FAP-specific substrate Suc-Gly-Pro-AMC. The assay enabled the fast determination of multiple IC50s by simply incubating a solution of phosphate-buffered saline in a 96-well plate within 30 min. The substrate specificity, cross-reaction and other related conditions were systematically optimized. This method was successfully applied to determine the IC50s of seven known inhibitors. The results are in consistence with the trend reported, which indicating that this practical assay is a valuable method to accelerate the discovery of FAP inhibitor.

Keywords: FAP inhibitor; Fibroblast activation protein; Fluorescent assay; Suc-Gly-Pro-AMC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Dose-Response Relationship, Drug
  • Drug Discovery*
  • Endopeptidases
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / pharmacology*
  • Gelatinases / antagonists & inhibitors*
  • Gelatinases / metabolism
  • Humans
  • Membrane Proteins / antagonists & inhibitors*
  • Membrane Proteins / metabolism
  • Microscopy, Confocal
  • Molecular Structure
  • Optical Imaging*
  • Serine Endopeptidases / metabolism
  • Structure-Activity Relationship

Substances

  • Fluorescent Dyes
  • Membrane Proteins
  • Endopeptidases
  • Serine Endopeptidases
  • fibroblast activation protein alpha
  • Gelatinases