Abstract
Non-steroidal anti-inflammatory drugs (NSAIDs) injure the proximal and distal gut by different mechanisms. While many drugs reduce gastrointestinal injury, no drug directly stimulates mucosal wound healing. Focal adhesion kinase (FAK), a non-receptor tyrosine kinase, induces epithelial sheet migration. We synthesized and evaluated a water-soluble FAK-activating small molecule, M64HCl, with drug-like properties. Monolayer wound closure and Western blots measured migration and FAK phosphorylation in Caco-2 cells, in vitro kinase assays established FAK activation, and pharmacologic tests assessed drug-like properties. 30 mg/kg/day M64HCl was administered in two murine small intestine injury models for 4 days. M64HCl (0.1–1000 nM) dose-dependently increased Caco-2 FAK-Tyr 397 phosphorylation, without activating Pyk2 and accelerated Caco-2 monolayer wound closure. M64HCl dose-responsively activates the FAK kinase domain vs. the non-salt M64, increasing the Vmax of ATP-binding. Pharmacologic tests suggested M64HCl has drug-like properties and is enterally absorbed. M64HCl 25 mg/kg/day continuous infusion promoted healing of ischemic jejunal ulcers and indomethacin-induced small intestinal injury in C57Bl/6 mice. M64HCl-treated mice exhibited smaller ulcers 4 days after ischemic ulcer induction or indomethacin injury. Renal histology and plasma creatinine were normal. Mild hepatic inflammatory changes and ALT elevation were similar among M64HCl-treated mice and controls. M64HCl was concentrated in kidney and gastrointestinal mucosa and functional nephrectomy studies suggested predominantly urinary excretion. Little toxicity was observed in vitro or in single-dose mouse toxicity studies until >1000x higher than effective concentrations. M64HCl, a water-soluble FAK activator, promotes epithelial restitution and intestinal mucosal healing and may be useful to treat gut mucosal injury.
Original language | English (US) |
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Article number | 100147 |
Journal | Current Research in Pharmacology and Drug Discovery |
Volume | 4 |
DOIs | |
State | Published - Jan 2023 |
Bibliographical note
Funding Information:Histological services were provided by the UND Histology Core Facility supported by NIH / NIGMS awards P20GM113123 , U54GM128729 , and UND SMHS funds. Supported in part by NIH 3UTGM13-175-02S1 and NIH U01HL152410 .
Funding Information:
The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:The University of North Dakota and the University of Minnesota have filed two patent applications addressing the use of small molecule FAK activators in promoting mucosal healing. Drs. Basson and Gurvich and Gallardo-Macias are named as inventors in at least one of each of these patents. The authors have no other potential competing interests to declare.Histological services were provided by the UND Histology Core Facility supported by NIH/NIGMS awards P20GM113123, U54GM128729, and UND SMHS funds. Supported in part by NIH 3UTGM13-175-02S1 and NIH U01HL152410. Marc D. Basson and Vadim J Gurvich are co-inventors of two patent applications currently pending that address the use of small molecule FAK activators to promote mucosal healing.
Publisher Copyright:
© 2022 The Authors
Keywords
- Focal adhesion kinase
- Mucosal healing
- Non-steroidal anti-inflammatory drugs
- Small intestine
- Ulcer