Potent and Selective Mechanism-Based Inhibition of Gelatinases

Brown, Stephen H.M.; Bernardo, M. Margarida; Li, § Zhi-Hong; Kotra, Lakshmi P.; Tanaka, Yasuhiro; Fridman, and Rafael; Mobashery, Shahriar

doi:10.1021/ja001461n

Cited by 192 publications

(248 citation statements)

References 17 publications

Supporting

Mentioning

239

Contrasting

Unclassified

Order By: Relevance

“…Marimastat (BB-2516) was obtained from British Biotech (Annapolis, MD) (35). SB-3CT was produced and characterized as described (36). The anti-TIMP-2 monoclonal antibody (mAb) CA-101 (33), the rabbit polyclonal antibody (pAb) 437 to the hemopexin-like domain of MT1-MMP (17), and the rabbit pAb to human TIMP-1 have been previously described (37,38).…”

Section: Methodsmentioning

confidence: 99%

Complex Pattern of Membrane Type 1 Matrix Metalloproteinase Shedding

Tóth¹,

Hernandez-Barrantes²,

Osenkowski³

et al. 2002

Journal of Biological Chemistry

Self Cite

112

View full text Add to dashboard Cite

Membrane type 1 matrix metalloproteinase (MT1-MMP 255 and Ile 256 residues near the conserved methionine turn, a structural feature of the catalytic domain of all MMPs. Consistently, a recombinant 18-kDa fragment had no catalytic activity and did not bind TIMP-2. Thus, autocatalytic shedding evolved as a specific mechanism to terminate MT1-MMP activity on the cell surface by disrupting enzyme integrity at a vital structural site. In contrast, functional data suggest that the non-autocatalytic shedding generates soluble active MT1-MMP species capable of binding TIMP-2. These studies suggest that ectodomain shedding regulates the pericellular and extracellular activities of MT1-MMP through a delicate balance of active and inactive enzyme-soluble fragments.Release of the extracellular portion of type I transmembrane proteins, referred to as ectodomain shedding, has been established as a major regulatory mechanism to control the activity of a variety of membrane-bound proteins on the cell surface (1). Recent evidence suggests that ectodomain shedding is also characteristic of the membrane type matrix metalloproteinases (MT-MMPs), 1 a subfamily of membrane-anchored MMPs by means of a transmembrane domain or a glycosylphosphatidylinositol anchor (2, 3). The MT-MMPs are major mediators of proteolytic events on the cell surface, including turnover of extracellular matrix components (4, 5), cleavage of various surface adhesion receptors (6 -8), and initiation of zymogen activation cascades (9, 10). Uncontrolled MT-MMP activity contributes to abnormal development (11) and is a key determinant in cancer metastasis and tumor angiogenesis (12)(13)(14). To control the extent of pericellular activity, the MT-MMPs are inhibited by the tissue inhibitors of metalloproteinases (TIMPs), a family of natural protein MMP inhibitors. In addition, MT-MMPs have a unique regulatory mechanism in which the active enzyme undergoes a series of processing steps, either autocatalytic (15-17) or mediated by other proteases (18), that regulate the activity and nature of the enzyme species at the cell surface and at the pericellular space. Previous studies have shown that active MT1-MMP is autocatalytically processed on the cell surface to an inactive membrane-tethered ϳ44-kDa species lacking the entire catalytic domain (17). This processing is inhibited by TIMP-2, TIMP-4, and synthetic MMP inhibitors consistent with being an intermolecular autocatalytic event (19,20). Inhibition of MT1-MMP processing induces accumulation of the active enzyme on the cell surface, and, as a consequence, net MT1-MMP-dependent proteolysis is enhanced. Indeed, we have shown that, under certain conditions, inhibition of MT1-MMP autocatalysis by synthetic MMP inhibitors enhances pro-MMP-2 activation by MT1-MMP in the presence of TIMP-2 (20, 21). Thus, although the presence of inhibitors will stabilize MT1-MMP on the cell surface, the absence or reduced levels of inhibitors will facilitate autocatalysis. As a membrane-anchored protein, the autocatalytic processing of ac...

show abstract

Section: Methodsmentioning

confidence: 99%

Complex Pattern of Membrane Type 1 Matrix Metalloproteinase Shedding

Tóth¹,

Hernandez-Barrantes²,

Osenkowski³

et al. 2002

Journal of Biological Chemistry

Self Cite

112

View full text Add to dashboard Cite

show abstract

“…We reported the first mechanism-based inhibitor (inhibitor 3) for MMPs, and initially we had targeted gelatinases due to their well-established role in cancer metastasis and angiogenesis. The dithiol inhibitors were designed based on the knowledge gained on the binding mode of the thiirane mechanism-based inhibitor 3 (40). The biphenyl moieties of 1, 2, and 3 are identical and this entity is designed to fit the S1Ј subsite of MMP-2.…”

Section: Fitmentioning

confidence: 99%

Structural Basis for Potent Slow Binding Inhibition of Human Matrix Metalloproteinase-2 (MMP-2)

Rosenblum

Meroueh

Kleifeld

et al. 2003

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

The zinc-dependent gelatinases belong to the family of matrix metalloproteinases (MMPs), enzymes that have been shown to play a key role in angiogenesis and tumor metastasis. These enzymes are capable of hydrolyzing extracellular matrix (ECM) components under physiological conditions. Specific and selective inhibitors aimed at blocking their activity are highly sought for use as potential therapeutic agents. We report herein on a novel mode of inhibition of gelatinase A (MMP-2) by the recently characterized inhibitors 4-(4-phenoxphenylsulfonyl)butane-1,2-dithiol (inhibitor 1) and 5-(4-phenoxphenylsulfonyl) pentane-1,2-dithiol (inhibitor 2). These synthetic inhibitors are selective for MMP-2 and MMP-9. We show that the dithiolate moiety of these inhibitors chelates the catalytic zinc ion of MMP-2 via two sulfur atoms. This mode of binding results in alternation of the coordination number of the metal ion and the induction of conformational changes at the microenvironment of the catalytic zinc ion; a set of events that is likely to be at the root of the potent slow binding inhibition behavior exhibited by these inhibitors. This study demonstrates a distinct approach for the understanding of the structural mechanism governing the molecular interactions between potent inhibitors and catalytic sites of MMPs, which may aid in the design of effective inhibitors.Matrix metalloproteinases (MMPs) 1 are major players in degradation of the extracellular matrix (ECM) components, and therefore they play key roles in normal and pathological conditions involving remodeling and turnover of ECM (1). MMPs constitute a multigene family of at least 26 secreted and membrane-tethered zinc-dependent endopeptidases, which can be classified according to their structures and substrate specificities (2, 3). Members of the family include: collagenases (MMP-1, -8, -13), gelatinases (MMP-2, -9), stromelysins , and membrane-type (MT)-MMPs (MT1-MMP to MT6-MMP). The catalytic domains of the MMPs have an ellipsoid shape with a small active site cleft. This cleft contains the catalytic zinc ion, which is essential for catalysis (4).Gelatinases, MMP-2 (gelatinase A) and MMP-9 (gelatinase B), constitute a distinct subgroup of the MMPs family due to the incorporation of three repeats of the fibronectin-type II module in the catalytic domain. These enzymes have been shown to play a key role in tumor cell invasion, metastasis, and angiogenesis by promoting degradation of ECM and the processing of cytokines, growth factors, hormones, and cell receptors (5, 6). As with other MMPs, MMP-2 and MMP-9 are expressed as zymogenic latent enzymes (pro-MMPs) requiring activation. Both proteolytic and non-proteolytic mechanisms have been described for zymogen activation. However, the "cysteine-switch" hypothesis (7) proposes that upon activation, the zinc ion in the latent active site is converted to a catalytic zinc ion by the dissociation of the conserved cysteine thiolate within the active site. The cleavage or the dissociation of the propeptide makes available ...

show abstract

“…NMR spectra were recorded in CDCl 3 on a Varian Inova 500 MHz, 400 MHz, or Varian Mercury 300 MHz spectrometer. 13 C NMR was recorded using broad band proton decoupling. Chemical shifts are reported in relative to TMS and coupling constants in Hz.…”

Section: General Experimental Procedures 16mentioning

confidence: 99%

“…Fluorine substitution on the o-position (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15) showed moderate inhibition for MMPs. Fluorine substitution on both m- (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16) and p-positions (3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17) showed good inhibition for MMP-1 (28, 23 µM). We also tested other fluorinated compounds and in particular, 3-21 displayed the best inhibition toward MMP-1 among all of inhibitors tested (13 µM).…”

mentioning

confidence: 99%

Synthesis and evaluation of novel heterocyclic MMP inhibitors

Hayashi

Jin

Cook

2007

Bioorganic & Medicinal Chemistry Letters

View full text Add to dashboard Cite

A variety of novel heterocyclic compounds were synthesized and evaluated for MMP inhibition. Broad spectrum inhibition of MMPs 1, 2, 9 and 12 was found with pyridinone-based compounds while N-heterocyclic triazoles and tetrazoles were largely ineffective. A highly selective tetrazole inhibitor for MMP-2 was discovered.Extracellular proteolysis plays a key role in many biological processes. Angiogenesis, wound healing, inflammatory reactions, management of the blood brain barrier, general maintenance of joints, organ development, ovulation, fetus implantation in the uterus, embryogenesis, and many others all depend on enzymes which remodel connective tissues in the extracellular matrix. A family of enzymes known as the matrix metalloproteinases (MMPs) are, at least in part, responsible for these functions. The important role of MMPs in extracellular degradation has been clearly demonstrated in many diseases including arthritis, multiple sclerosis, osteoporosis, Alzheimer's disease, cancer growth and metastasis, and periodontal disease. 1 For example, MMPs mediate extracellular matrix and basement-membrane degradation during the early stages of tumor growth particularly the gelatinases. 2 Abnormal levels of MMPs are observed for patients of these diseases and thus inhibition of MMPs offers a potential for new therapeutics. There are more than 24 known MMPs. The MMP family of enzymes is structurally related with a zinc-containing catalytic site. There are metalloproteinases that utilize a zinc (II) metal for catalysis of the proteolysis, and therefore, zinc binding groups (ZBGs) are crucial for designing MMP inhibitors (MMPIs). As a consequence, there have been a great deal of effort in recent years to design and prepare inhibitors of MMPs, mostly targeted at the prime side of the active site, which contains a hydrophobic S1' pocket (figure 1a). Key to the activity of almost all MMPIs is the functional group that binds the zinc atom in the sctive site and the P1' substituent. 3,4 To date, the only medically approved MMPI is a tetracycline derivatives with µM range inhibitions used in the treatment of periodontal disease. In the last few decades, hydroxamate based MMPIs has been mainly studied and fair amount of structureactivity data is available on these compound. Many small molecule MMPIs with hydroxamic acid as a ZBG have shown outstanding inhibition in preclinical tests. Moreover, MMPIs with other ZBGS such as thiols, carboxylates, mercaptoalcohols, 5 and dithiols, have shown good inhibition in vitro. 6 However, clinical tests for these compounds have been disappointing. Some of the reasons for this include low oral availability, poor in vivo stability, pharmacokinetic problems, 7 and undesirable side effects associated with hydroxamates. Therefore the discovery of new MMPIs with better properties and non-toxic ZBGs is critical. Recent reports demonstrate a continued optimism that MMPI therapy might be beneficial. 8-Gregory.Cook@ndsu.edu. Publisher's Disclaimer: This is a PDF file of an unedited manuscr...

show abstract

Potent and Selective Mechanism-Based Inhibition of Gelatinases

Cited by 192 publications

References 17 publications

Complex Pattern of Membrane Type 1 Matrix Metalloproteinase Shedding

Complex Pattern of Membrane Type 1 Matrix Metalloproteinase Shedding

Structural Basis for Potent Slow Binding Inhibition of Human Matrix Metalloproteinase-2 (MMP-2)

Synthesis and evaluation of novel heterocyclic MMP inhibitors

Contact Info

Product

Resources

About