Peptide inhibitors of MK2 show promise for inhibition of abdominal adhesions

J Surg Res. 2011 Jul;169(1):e27-36. doi: 10.1016/j.jss.2011.01.043. Epub 2011 Feb 23.

Abstract

Background: Abdominal adhesions are a common side effect of surgical procedures with complications including infertility, chronic pain, and bowel obstruction, which may lead to the need for surgical lyses of the adhesions. Mitogen-activated protein kinase-activated protein kinase 2 (MK2) has been implicated in several diseases, involving inflammation and fibrosis. Thus, the development of a cell-penetrating peptide (CPP) that modulates MK2 activity may confer therapeutic benefit after abdominal surgery in general and more specifically after bowel anastomosis.

Methods: This study evaluated the function of a CPP inhibitor of MK2 in human mesothelial cells and in a rat bowel anastomosis model. To determine IC50 and basic specificity, kinase inhibition was performed using a radiometric assay. Enzyme-linked immunoassay (ELISA) was used to evaluate interleukin-6 (IL-6) expression in response to IL-1β and tumor necrosis factor-α (TNF-α) stimulation in vitro to validate MK2 kinase inhibition. Following bowel anastomosis (10 rats for each control and treatment at 4 and 10 d), the rats were evaluated for weight loss, normal healing (colonic burst strength and hydroxyproline content at the anastomosis), and number and density of adhesions.

Results: The IC50 of the MK2 inhibitor peptide (22 μM) was similar to that of the nonspecific small molecule rottlerin (IC50 = 5 μM). The MK2 inhibitor peptide was effective at suppressing IL-1β and TNF-α stimulated IL-6 expression in mesothelial cells. In vivo, the MK2 inhibitor peptide was effective at suppressing both the density and number of adhesions formed as a result of bowel an anastamosis. Importantly, the peptide had no negative effect on normal healing.

Conclusions: In conclusion, the peptide inhibitor of MK2, MMI-0100, has the potential to significantly reduce inflammation through suppression of inflammatory cytokine expression and showed promise as a therapeutic for abdominal adhesions.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Abdomen / surgery*
  • Animals
  • Cell-Penetrating Peptides / pharmacology
  • Cell-Penetrating Peptides / therapeutic use*
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Humans
  • Hydroxyproline / metabolism
  • Inhibitory Concentration 50
  • Interleukin-1beta / pharmacology
  • Interleukin-6 / metabolism
  • Intracellular Signaling Peptides and Proteins / antagonists & inhibitors*
  • Intracellular Signaling Peptides and Proteins / physiology
  • Models, Animal
  • Protein Kinase Inhibitors / pharmacology
  • Protein Kinase Inhibitors / therapeutic use*
  • Protein Serine-Threonine Kinases / antagonists & inhibitors*
  • Protein Serine-Threonine Kinases / physiology
  • Rats
  • Time Factors
  • Tissue Adhesions / metabolism
  • Tissue Adhesions / physiopathology
  • Tissue Adhesions / prevention & control*
  • Tumor Necrosis Factor-alpha / pharmacology

Substances

  • Cell-Penetrating Peptides
  • Interleukin-1beta
  • Interleukin-6
  • Intracellular Signaling Peptides and Proteins
  • Protein Kinase Inhibitors
  • Tumor Necrosis Factor-alpha
  • MAP-kinase-activated kinase 2
  • Protein Serine-Threonine Kinases
  • Hydroxyproline