Small molecule FAK activator promotes human intestinal epithelial monolayer wound closure and mouse ulcer healing

Sci Rep. 2019 Oct 11;9(1):14669. doi: 10.1038/s41598-019-51183-z.

Abstract

GI mucosal healing requires epithelial sheet migration. The non-receptor tyrosine kinase focal adhesion kinase (FAK) stimulates epithelial motility. A virtual screen identified the small drug-like FAK mimic ZINC40099027, which activates FAK. We assessed whether ZINC40099027 promotes FAK-Tyr-397 phosphorylation and wound healing in Caco-2 monolayers and two mouse intestinal injury models. Murine small bowel ulcers were generated by topical serosal acetic acid or subcutaneous indomethacin in C57BL/6J mice. One day later, we began treatment with ZINC40099027 or DMSO, staining the mucosa for phosphorylated FAK and Ki-67 and measuring mucosal ulcer area, serum creatinine, ALT, and body weight at day 4. ZINC40099027 (10-1000 nM) dose-dependently activated FAK phosphorylation, without activating Pyk2-Tyr-402 or Src-Tyr-419. ZINC40099027 did not stimulate proliferation, and stimulated wound closure independently of proliferation. The FAK inhibitor PF-573228 prevented ZINC40099027-stimulated wound closure. In both mouse ulcer models, ZINC40099027accelerated mucosal wound healing. FAK phosphorylation was increased in jejunal epithelium at the ulcer edge, and Ki-67 staining was unchanged in jejunal mucosa. ZINC40099027 serum concentration at sacrifice resembled the effective concentration in vitro. Weight, creatinine and ALT did not differ between groups. Small molecule FAK activators can specifically promote epithelial restitution and mucosal healing and may be useful to treat gut mucosal injury.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Caco-2 Cells
  • Cell Movement / drug effects
  • Epithelial Cells / drug effects*
  • Epithelial Cells / pathology
  • Focal Adhesion Protein-Tyrosine Kinases / antagonists & inhibitors
  • Focal Adhesion Protein-Tyrosine Kinases / genetics*
  • Humans
  • Intestinal Mucosa / drug effects*
  • Intestinal Mucosa / pathology
  • Jejunum / drug effects
  • Jejunum / pathology
  • Mice
  • Mice, Inbred C57BL
  • Phosphorylation / drug effects
  • Quinolones / pharmacology
  • Small Molecule Libraries / pharmacology
  • Sulfones / pharmacology
  • Ulcer / drug therapy*
  • Ulcer / genetics
  • Ulcer / pathology
  • Wound Closure Techniques
  • Wound Healing / drug effects
  • Wound Healing / genetics

Substances

  • 6-(4-(3-(methylsulfonyl)benzylamino)-5-(trifluoromethyl)pyrimidin-2-ylamino)-3,4-dihydroquinolin-2(1H)-one
  • Quinolones
  • Small Molecule Libraries
  • Sulfones
  • Focal Adhesion Protein-Tyrosine Kinases