Development of Matrix Metalloproteinase-2 Inhibitors for Cardioprotection

Abstract

The objective of our present study is to develop novel inhibitors for MMP-2 for acute cardioprotection. In a series of pilot studies, novel substituted carboxylic acid derivatives were synthesized based on imidazole and thiazole scaffolds and then tested in a screeening cascade for MMP inhibition. We found that the MMP-inhibiting effects of imidazole and thiazole carboxylic acid-based compounds are superior in efficacy in comparison to the conventional hydroxamic acid derivatives of the same molecules. Based on these results, a 568-membered focused library of imidazole and thiazole compounds was generated in silico and then the library members were docked to the 3D model of MMP-2 followed by an in vitro medium throughput screening (MTS) based on a fluorescent assay employing MMP-2 catalytic domain. Altogether 45 compounds showed a docking score of >70, from which 30 compounds were successfully synthesized. Based on the MMP-2 inhibitory tests using gelatin zymography, 7 compounds were then selected and tested in neonatal rat cardiac myocytes subjected to simulated I/R injury. Six compounds showed significant cardio-cytoprotecion and the most effective compound (MMPI-1154) significantly decreased infarct size when applied at 1 μM in an ex vivo model for acute myocardial infarction. This is the first demonstration that imidazole and thiazole carboxylic acid-based compounds are more efficacious MMP-2 inhibitor than their hydroxamic acid derivatives. MMPI-1154 is a promising novel cardio-cytoprotective imidazole-carboxylic acid MMP-2 inhibitor lead candidate for the treatment of acute myocardial infarction.

Cardioprotective Effect of Novel Matrix Metalloproteinase Inhibitors

“…MMPI-1154, -1260, and -1248 inhibitors were shown to be cardiocytoprotective in in vitro cell culture experiments, as we previously published [ 27 ], and here we tested their efficacy in vivo.…”

“…According to Jacobsen et al, to achieve highly specific MMP inhibitors, hydroxamate and carboxylate zinc-binding groups could be developed by careful consideration of inhibitor backbones for targeting the different substrate pockets of individual MMPs [ 20 ]. Our attempts to increase the selectivity for MMP-2 against MMP-1 and other MMPs were embedded in the pyridine moiety instead of the phenyl ring at the end of the S1′ pocket, occupying a longer side chain of the novel inhibitor candidates (MMPI-1260, 1248) [ 27 ]. Another important aspect during inhibitor screening was to inhibit MMP-2 activity moderately, which may be accompanied by limited side effects [ 35 , 38 ].…”

“…Our molecules were developed through a screening cascade from the in silico Albany Molecular Ressearch Inc. (AMRI) Chemical Library to find zinc-binding motif-holding molecules similar to hydroxamic acids. Thiazole and isosteric imidazole carboxylic acids were chosen and screened through different in vitro assays in order to select the most potent MMP inhibitors for in vivo experiments aiming for cardioprotection (for details, see [ 27 ]).…”

“…In the present study, three selected imidazole (MMPI-1154) and thiazole (MMPI-1248 and -1260) carboxylic acid-type MMP-2 inhibitors were tested in an in vivo rat model of AMI. MMPI-1154, -1260 and -1248 were tested in three different doses based on previous half maximal inhibitory concentration (IC 50 ) values (IC 50 for MMP-2 MMPI-1154: 2.5 µM, MMPI-1260: 2.6 µM and MMPI-1248: 9 µM) [ 27 ]. MMPI-1154 at 1 µmol/kg and MMPI-1260 at 3 µmol/kg showed cardioprotection by significantly lowering infarct size compared to the vehicle-treated group in normocholesterolemic rats subjected to AMI.…”

“…These imidazole and thiazole carboxylic acid-based compounds showed superior MMP-inhibiting effects when compared to the hydroxamic acid-type derivatives in vitro in gelatin zymography assays. Six compounds, including MMP inhibitors (MMPI) 1154, MMPI-1260 and MMPI-1248 were found to be protective against simulated ischemia/reperfusion induced injury in cardiac myocytes [ 27 ]. Here, we further test these lead molecular compounds in vivo in relation to acute myocardial infarction in normo- and hypercholesterolemic animals, as hypercholesterolemia is one of the most frequent co-morbidities of acute myocardial infarction.…”