Development of Selective Inhibitors and Substrate of Matrix Metalloproteinase-12

Abstract

The matrix metalloproteinases (MMPs)2 form a group of structurally related extracellular zinc endoproteases able to degrade at least one protein component of the extracellular matrix (1). Based on this property, MMPs are considered to be critical mediators of both normal and pathological tissue remodeling processes (2, 3). Their overexpression is observed in and associated with a variety of diseases, including cancer (4, 5), arthritis (6), multiple sclerosis (7,8), and atherosclerosis (9, 10). Therefore, there is substantial interest in developing MMP synthetic inhibitors for a variety of therapeutic indications (11)(12)(13)(14)(15). Results of the first clinical trials with broad spectrum MMP inhibitors in cancer therapy were disappointing, highlighting the need for better understanding of the exact role of each MMP during the different stages of tumor progression (16). Recent research in this field has focused on the development of inhibitors that fully differentiate one MMP from another (17). This is a particularly difficult task, since the topology and nature of the residues in the enzyme's active site are highly conserved among the different MMPs (18). Moreover, parts of the MMP catalytic domain, which play a critical role in enzyme specificity, seem to be highly flexible (19,20). This situation may explain why most previously reported MMP synthetic inhibitors preferentially inhibit some MMPs but are not exclusive inhibitors of a single MMP. A recent breakthrough in this field was achieved by identifying the first highly selective synthetic inhibitor of MMP-13 (21). Selective inhibitors for MMP-2 and MMP-9 have also been reported recently, but their degree of selectivity toward MMPs is less than that achieved for the MMP-13-selective inhibitor (22).To identify highly selective MMP inhibitors, libraries of phosphinic peptides were prepared. Phosphinic peptides are good transition state mimics and have been shown to behave as highly potent inhibitors of different zinc metalloproteinases (23). To probe the SЈ 1 cavity of MMPs, a chemical strategy that makes it possible to prepare phosphinic peptides harboring various substituents in their PЈ 1 position was used (see Scheme 1). This strategy relies on the use of a common precursor, which can be modified in one step, to prepare phosphinic peptides displaying substituted isoxazole side chains in their PЈ 1 position (24). In such phosphinic peptides, the isoxazole ring is used as a rigid scaffold to project in the right orientation various chemical groups able to interact with the SЈ 1 subsite of MMPs, which corresponds to a deep cavity. Based on this strategy, four libraries of phosphinic peptides, containing four different isoxazole side chains in their PЈ 1 position, were prepared by introducing additional chemical diversity in the PЈ 2 and PЈ 3 positions of the inhibitors (see Scheme 1). Screening of these libraries against 10 different MMPs allowed us to identify highly selective inhibitors of MMP-12. Based on the unique structure of these inhibitors, a highl...

Upregulation of elastase proteins results in aortic dilatation in mucopolysaccharidosis I mice

“…Indeed, MMPs contributed to joint disease in other MPS models [44] while cathepsin S was upregulated in the brain of MPS I mice [45]. If the genetic cross that is in progress shows that deficiency of one or both of these elastase proteins can reduce aortic disease in MPS I, it might be possible to treat MPS I patients with inhibitors of MMP-12 [46] and/or cathepsin S [47]. Evaluation of transcription factor mRNA in aorta.…”

Discovery of a New Class of Potent MMP Inhibitors by Structure-Based Optimization of the Arylsulfonamide Scaffold

“…Several potent inhibitors for MMP-12, such as phospinic peptides, sulfonamides, and bisphosphonic derivatives, have already been designed and tested. 5−10 For some of them a high selectivity toward MMP-12 has been observed, 5,6,8,10 although the molecular basis of this specificity is still a matter of research. Here, based on the classical arylsulfonamide scaffold, new MMP inhibitors were designed by introducing an ethylene linker between the sulfonamide moiety and the P1′ aromatic portion and by replacing the glycine residue with a D-proline within the zinc-binding group (ZBG).…”

Solution Structure of Inhibitor-Free Human Metalloelastase (MMP-12) Indicates an Internal Conformational Adjustment

“…15 N and 15 N, 13 C labelled proteins were expressed from a pGEMEX expression plasmid (Promega) in an E. coli BL21(DE3) RIL host using 15 NH 4 CI and 13 Cglucose-based M9 medium supplemented with Celtone (Spectral Stable Isotopes) at 20% (v/ v). The expressed protein is found in inclusion bodies.…”