Inhibitory effects of caspase inhibitors on the activity of matrix metalloproteinase-2.

Abstract

Matrix metalloproteinase (MMP)-2, a zinc-dependent endopeptidase, plays a detrimental role in several diseases including ischemia and reperfusion (I/R) injury of the heart. Caspases are a group of cysteine-dependent, aspartate-directed proteases which regulate cellular apoptosis. Interestingly, protective effects of caspase inhibitors independent of apoptosis have been shown in I/R injury of the heart. The cardioprotective actions of both these classes of protease inhibitors led us to hypothesize that caspase inhibitors may also reduce MMP-2 activity. Five known caspase inhibitors (Z-IE(OMe)TD(OMe)-fmk, Ac-DEVD-CHO, Ac-LEHD-cmk, Z-VAD-fmk and Ac-YVAD-cmk) were tested for their possible inhibitory effects on MMP-2 activity in comparison to the MMP inhibitors ONO-4817 and ARP-100 (which themselves were unable to inhibit caspase-3 activity). MMP-2 activity was assessed by an in vitro troponin I (TnI) proteolysis assay and a quantitative kinetic fluorescence assay using a fluorogenic peptide substrate (OmniMMP). TnI proteolysis was also measured by western blot in neonatal cardiomyocytes subjected to hypoxia-reoxygenation injury. Using human recombinant MMP-2 and TnI as its substrate, the caspase inhibitors, in comparison with ONO-4817, significantly inhibited MMP-2-mediated TnI degradation in a concentration-dependent manner. The kinetic assay using OmniMMP revealed that these caspase inhibitors blocked MMP-2 activity in a concentration-dependent manner with similar IC50 values. TnI degradation in neonatal cardiomyocytes was enhanced following hypoxia-reoxygenation and this was blocked by ARP-100 and Ac-LEHD-cmk. Inhibition of MMP-2 activity is an additional pharmacological action which contributes to the protective effects of some caspase inhibitors.

• Authors: M M Castro, J Fuah, M Ali, M Sung, J Schulz, M Y Kondo, X Fan, A Holt, R Schulz
• Journal: Biochemical pharmacology (Biochem Pharmacol) Vol. 86 Issue 4 Pg. 469-75 (Aug 15 2013) ISSN: 1873-2968 [Electronic] England
• PMID: 23774623 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2013 Elsevier Inc. All rights reserved.

Chemical References

• Caspase Inhibitors
• Hydroxamic Acids
• Matrix Metalloproteinase Inhibitors
• N-hydroxy-2-((4-phenylphenyl)sulfonylpropan-2-yloxyamino)acetamide
• N-hydroxy-5-ethoxymethyloxy-2-methyl-4-(4-phenoxybenzoyl)aminopentanamide
• Oligopeptides
• Phenyl Ethers
• Sulfones
• Troponin I
• Caspase 3
• Matrix Metalloproteinase 2

Topics

• Animals
• Animals, Newborn
• Caspase 3 (metabolism)
• Caspase Inhibitors (pharmacology)
• Cell Hypoxia
• Cells, Cultured
• Humans
• Hydroxamic Acids (pharmacology)
• Matrix Metalloproteinase 2 (metabolism)
• Matrix Metalloproteinase Inhibitors (pharmacology)
• Myocytes, Cardiac (drug effects, enzymology)
• Oligopeptides (pharmacology)
• Phenyl Ethers (pharmacology)
• Rats
• Rats, Sprague-Dawley
• Sulfones (pharmacology)
• Troponin I (metabolism)