Molecular Determinants of a Selective Matrix Metalloprotease-12 Inhibitor: Insights from Crystallography and Thermodynamic Studies

Czarny, Bertrand; Stura, E.A.; Devel, Laurent; Vera, Laura; Cassar-Lajeunesse, E.; Beau, Fabrice; Calderone, V.; Fragai, Marco; Luchinat, Claudio; Dive, Vincent

doi:10.1021/jm301574d

Cited by 34 publications

(53 citation statements)

References 54 publications

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“…The phosphinic pseudo peptide part of 1 binds to MMP-12 in a comparable manner to that observed for other phosphinic derivatives in interaction with this protease 30 . In this respect, the two oxygen atoms of the phosphoryl function are not placed symmetrically around the catalytic zinc ion and only one oxygen binds to this atom (Figure 2A and 2C).…”

Section: Resultssupporting

confidence: 62%

“…The probe’s design was based on the crystallographic structure of RXP470.1 in complex with MMP-12 30 . The essential roles played by the RXP470.1 hydrophobic P 1 ′ side chain as well as the two glutamate residues at the P 2 ′ and P 3 ′ positions in preserving the strong affinity and selectivity in favor of MMP-12 were taken into account 30 . We reasoned that a chemical elongation at the C -terminal position, should have a negligible impact on the binding of RXP470.1 motif within the active site and that steric clashes or long-distance electrostatic effects could be prevented by moving away the fluorophore from the MMP-12 targeting moiety using a polyethylene glycol (PEG) spacer.…”

Section: Resultsmentioning

confidence: 99%

“…Several X-ray diffraction data sets were collected at the synchrotron Soleil beamlines Proxima 1 and 2A to determine the positioning of the fluorophore moiety within the crystal lattice of MMP-12 molecules. In the crystallization, the MMP-12 mutant F171D reported for the crystallization of RXP470.1 30 and a second mutant with the additional K241A mutation were used. Both gave good quality diffracting crystals in two different polymorphs (Supporting Information, Table S1), a new monoclinic, P2 1 crystal form with two molecules in the asymmetric unit, never obtained before 38 , and the classical orthorhombic, P2 1 2 1 2 crystal form 30,39,40 with one molecule in the asymmetric unit (Supporting Information, Table S1).…”

Section: Resultsmentioning

confidence: 99%

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Synthesis and in Vitro and in Vivo Evaluation of MMP-12 Selective Optical Probes

et al. 2016

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In designing new tracers consisting of a small-peptide conjugated to a reporter of comparable size, particular attention needs to be paid to the selection of the reporter group which can dictate both the in vitro and in vivo performances of the whole conjugate. In the case of fluorescent tracers, this is particularly true given the large numbers of available dye moieties differing in their structures and properties. Here, we have investigated the in vitro and in vivo properties of a novel series of MMP-12 selective probes composed of cyanine dyes varying in their structure, net charge and hydrophilic character, tethered through a linker to a potent and specific MMP-12 phosphinic pseudo peptide inhibitor. The impact of linker length has been also explored. The crystallographic structure of one tracer in complex with MMP-12 has been obtained, providing the first crystal structure of a Cy5.5-derived probe and confirming that the binding of the targeting moiety is unaffected. MMP-12 remains the tracers privileged target as attested by their affinity selectivity profile evaluated in solution towards a panel of twelve metalloproteases. In vivo assessment of four selected probes has highlighted the impact of the dye structure, but also that of the linker length on the probes blood clearance rates and their biodistributions. These experiments have also provided valuable data on the stability of the dyes moieties in vivo. This has permitted the identification of one probe, which combines favorable binding to MMP-12 in solution and on cells with optimized in vivo performance including blood clearance rate suitable for short-time imaging. Through this series of tracers, we have identified various critical factors modulating the tracers’ in vivo behavior, both useful for the development and optimization of MMP-12 selective radiolabeled tracers and informative for the design of fluorescent probes in general.

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Section: Resultssupporting

confidence: 62%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Synthesis and in Vitro and in Vivo Evaluation of MMP-12 Selective Optical Probes

et al. 2016

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“…147,148 Pseudopeptides with the general formula X-l-Glu-NH 2 that affected zinc ion binding were recognized as MMP12 inhibitors. 149 More recently, two monoclonal antibodies have been designed to inhibit substrate activation by binding to the catalytic domain without affecting the catalytic zinc region. 38 The first, DX2400, is a specific inhibitor of MMP14, known to activate pro-MMP2, and promotes angiogenesis, cell invasion, and metastasis in breast cancer cells.…”

mentioning

confidence: 99%

Matrix metalloproteinases in head and neck cancer: current perspectives

Gkouveris¹,

Nikitakis²,

Aseervatham³

et al. 2017

MNM

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Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases encoded by 24 distinct genes. Their functions have been implicated in numerous normal and pathologic processes, including uterine involution and organogenesis, inflammation and wound healing, vascular and autoimmune disease progression. Pertinent to this review, the role of MMPs in cancer biology is fairly well researched and documented, and remains a subject of continuing intense investigation. Not only are several MMPs overexpressed in head and neck squamous cell carcinomas (HNSCCs), expression has been correlated with salient tumorigenic hallmarks, such as cell proliferation, angiogenesis, invasion, and metastasis. The utility of changes in the expression profile, as well as various MMP polymorphisms as potential prognostic markers in oral cancers and oral premalignant lesions, have been investigated. Furthermore, the potential therapeutic utility of targeting MMPs in cancer remains attractive, although outcomes in this respect appear so far to be less encouraging with respect to HNSCCs. Because of the disappointing results observed in clinical trials where MMP-targeting regimens for HNSCCs utilized broad-spectrum small MMP catalytic site inhibitors, investigators now envision new strategies for MMP-specific targeting based on the recognition of new noncatalytic MMP domains with distinct functions. This review provides an overview of MMP activities in general and in cancers, and an update of their activities in HNSCC. Specifically, their role in the development and progression of HNSCC and their function as signaling molecules is discussed. Finally, their role as potential prognostic biomarkers and therapeutic targets in HNSCC is revisited.

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“…For MMP-12 in particular, enhanced understanding of the molecular determinants of drug affinity have been aided by numerous very high resolution crystal structures (134, 135). Novel and selective inhibitors of MMP-12 have resulted from further optimization of S1′ pocket fit for this enzyme (136), as well as from incorporating P2′ glutamate into pseudo-dipeptides, which in the absence of a traditional zinc-chelating group, can take on a non-canonical binding conformation in which it interacts with the catalytic zinc (137).…”

Section: Therapeutic Approaches Targeting Mmpsmentioning

confidence: 99%

Matrix metalloproteinases as drivers and therapeutic targets in breast cancer

2015

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Members of the matrix metalloproteinase (MMP) family have been identified as poor prognosis markers for breast cancer patients and as drivers of many facets of the tumor phenotype in experimental models. Early enthusiasm for MMPs as therapeutic targets was tempered following disappointing clinical trials that utilized broad spectrum, small molecule catalytic site inhibitors. However, subsequent research has continued to define key roles for MMPs as breast cancer promoters, to elucidate the complex roles that that these proteins play in breast cancer development and progression, and to identify how these roles are linked to specific and unique biochemical features of individual members of the MMP family. Here, we provide an overview of the structural features of the MMPs, then discuss clinical studies identifying which MMP family members are linked with breast cancer development and new experimental studies that reveal how these specific MMPs may play unique roles in the breast cancer microenvironment. We conclude with a discussion of the most promising avenues for development of therapeutic agents capable of targeting the tumor-promoting properties of MMPs.

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Molecular Determinants of a Selective Matrix Metalloprotease-12 Inhibitor: Insights from Crystallography and Thermodynamic Studies

Cited by 34 publications

References 54 publications

Synthesis and in Vitro and in Vivo Evaluation of MMP-12 Selective Optical Probes

Synthesis and in Vitro and in Vivo Evaluation of MMP-12 Selective Optical Probes

Matrix metalloproteinases in head and neck cancer: current perspectives

Matrix metalloproteinases as drivers and therapeutic targets in breast cancer

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