Discovery of CGS 27023A, a Non-Peptidic, Potent, and Orally Active Stromelysin Inhibitor That Blocks Cartilage Degradation in Rabbits

MacPherson, Lawrence; Bayburt, Erol K.; Capparelli, Michael; Carroll, Brian; Goldstein, Robert J.; Justice, Michael R.; Zhu, Lijuan; Hu, S; Melton, Richard; Fryer, L R; Goldberg, R.L.; Doughty, John; Spirito, S.; Blancuzzi, V.; Wilson, Doug; O’Byrne, E. M.; Ganu, Vishwas; Parker, David

doi:10.1021/jm960871c

Cited by 331 publications

(271 citation statements)

References 24 publications

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“…To evaluate the sustained expression of Wnt1 in our transfected clones, RT-PCR were performed using the following primers: 5'-CAT CGA GTC CTG CAC CTG-3'; 5'-TGG GCG ATT TCT CGA AGT AG-3'. 47 In some conditions, cells were cultured in the presence of the natural MMP inhibitor rTIMP-2 (10 µg/ ml), 48 the synthetic broad spectrum MMP inhibitors AG3340 (10 µg/ml) (Prinomastat, Agouron/Pfizer, Inc., San Diego, CA), or N-isobutyl-N-(4-methoxyphenylsulfonyl)glycyl hydroxamic acid (NNGH, 50 µM) 49,50 (Enzo Life Sciences, Inc., Plymouth Meeting, PA). Immortalized mouse fibroblast L cells and Wnt3a-expressing L cells (ATCC, Manassas, VA) were maintained in DMEM containing 10% FCS.…”

Section: Methodsmentioning

confidence: 99%

Stromelysin-1 (MMP-3) is a target and a regulator of Wnt1-induced epithelial-mesenchymal transition (EMT)

Blavier¹,

Lazaryev²,

Shi³

et al. 2010

Cancer Biology & Therapy

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Section: Methodsmentioning

confidence: 99%

Stromelysin-1 (MMP-3) is a target and a regulator of Wnt1-induced epithelial-mesenchymal transition (EMT)

Blavier¹,

Lazaryev²,

Shi³

et al. 2010

Cancer Biology & Therapy

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“…For inducible expression of wild-type or catalytically inactive MT1-MMP mutant (MT1 E/A) in A431 cells we used the RevTet-OffTM Gene Expression Systems (Clontech) according to the manufacturer's instructions. MMI270 (a synthetic hydroxamic MMP inhibitor, a kind gift of Novartis Pharma AG, Basel, Switzerland) (24) was added when the medium was changed 24 h after transfection.…”

Section: Methodsmentioning

confidence: 99%

Identification and Characterization of Lutheran Blood Group Glycoprotein as a New Substrate of Membrane-type 1 Matrix Metalloproteinase 1 (MT1-MMP)

Niiya

Egawa

Sakamoto

et al. 2009

Journal of Biological Chemistry

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Membrane-type 1 matrix metalloproteinase 1 (MT1-MMP) is a potent modulator of the pericellular microenvironment and regulates cellular functions in physiological and pathological settings in mammals. MT1-MMP mediates its biological effects through cleavage of specific substrate proteins. However, our knowledge of MT1-MMP substrates remains limited. To identify new substrates of MT1-MMP, we purified proteins associating with MT1-MMP in human epidermoid carcinoma A431 cells and analyzed them by mass spectrometry. We identified 163 proteins, including membrane proteins, cytoplasmic proteins, and functionally unknown proteins. Sixty-four membrane proteins were identified, and they included known MT1-MMP substrates. Of these, eighteen membrane proteins were selected, and we confirmed their association with MT1-MMP using an immunoprecipitation assay. Cells in tissues are surrounded by an extracellular cellular matrix that interacts with cells to regulate their activity (1, 2). Matrix metalloproteinases (MMPs) 3 are endopeptidases responsible for extracellular matrix degradation and thereby regulate turnover of the extracellular matrix. However, recent studies have demonstrated that substrates of MMPs are expanded to a variety of pericellular proteins. MT1-MMP/MMP14 is an integral membrane proteinase that cleaves multiple proteins in the pericellular milieu and thereby regulates various cell functions. Substrates of MT1-MMP identified to date include extracellular matrix proteins (type I collagen, fibronectin, vitronectin, laminin-1 and -5, and others), cell adhesion molecules (CD44, syndecan-1, and ␣v integrin), cytokines (SDF-1 and transforming growth factor-␤ and others), and latent forms of pro-MMPs (pro-MMP-2 and pro-MMP13) (3-5). Processing of these proteins by MT1-MMP alters their activities and thereby regulates a variety of cellular functions, such as motility, invasion, growth, differentiation, and apoptosis. Consistent with these functions, forced expression of MT1-MMP in tumor cells enhances behavior consistent with increased malignancy, such as rapid tumor growth, invasion, and metastasis (6). However, MT1-MMP is normally expressed in various types of cell and mice deficient in MT1-MMP expression (MT1 Ϫ/Ϫ ) display pleiotropic defects (7-10). However, we as yet have only limited knowledge of the physiological substrates of MT1-MMP that could explain such pleiotropic effects.Proteases interact with their substrates at least transiently, but in some cases such interaction is more stable. For instance, type I collagen binds MT1-MMP via a hemopexin-like domain and is cleaved (11, 12). Cleavage of collagen by MT1-MMP regulates cell growth and invasion in a collagen-rich environment (13). CD44, a hyaluronic acid receptor, also binds to the hemopexin of MT1-MMP and is cleaved (14). Expression of CD44 and MT1-MMP in tumor cells promotes cell migration, accompanied by the shedding of CD44 by MT1-MMP (14, 15). pro-MMP-2, which is cleaved by MT1-MMP for activation, forms a tri-molecular complex with MT1-MMP and TIM...

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“…Inhibitor 444225 was designed to be a potent deep S 1 Ј pocket inhibitor of MMP-3 (K i ϭ 130 nM; 47). The 4-methoxybenzenesulfonyl group of these inhibitors binds at the deep S 1 Ј pocket according to the crystallographic structure (45) and the structure-activity relationship of several derivatives (47) (19). Thus, homology modeling and protein sequence alignment may be useful tools to predict key residues involved in forming the S 1 Ј pocket of MMP-26.…”

Section: Inhibition Of Mmps With Mercaptosulfidementioning

confidence: 99%

The Intermediate S1′ Pocket of the Endometase/Matrilysin-2 Active Site Revealed by Enzyme Inhibition Kinetic Studies, Protein Sequence Analyses, and Homology Modeling

Park

Jin

Hurst

et al. 2003

Journal of Biological Chemistry

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Matrix metalloproteinases (MMPs, 1 matrixins) are believed to participate in angiogenesis, embryonic development, morphogenesis, reproduction, tissue resorption and remodeling, and tumor growth, progression, invasion, and metastasis through breakdown of the extracellular matrix, cell surface proteins, and processing growth factors, cytokines, and chemokines (1-3). Recently, human MMP-26 (endometase/matrilysin 2) was identified and its mRNA expression was detected in normal tissues of the human uterus and placenta, and in many types of malignant tumors (4 -7). Characterization of the MMP-26 promoter suggests that this proteinase may be expressed in cancer cells of epithelial origin (8). MMP-26 may play an important role in human prostate and breast cancer invasion (9 -10).MMP-26 cleaves type I gelatin, ␣ 1 -proteinase inhibitor, fibrinogen, fibronectin, vitronectin, type IV collagen, and insulin-like growth factor binding protein-1 (4, 7, 11). Studies of MMP-26 indicate that it has substrate specificity similar to other MMPs, with the exception of a preference for Ile at the P 2 and P 2 Ј positions, for small residues at the P 3 Ј and P 4 Ј positions, and Lys at the P 4 position (11). MMP-26 also hydrolyzes several synthetic fluorogenic peptide substrates designed for stromelysin-1, gelatinases, collagenases, and tumor necrosis factor-␣ converting enzyme (4, 11). According to these peptide substrate studies, MMP-26 may be capable of cleaving a broad range of substrates, although it has less catalytic efficiency than other MMPs.X-ray crystal structures of MMPs illustrate that overall topology and secondary structures are conserved (12-18). The S 1 Ј pocket, a hydrophobic pocket of variable depth, is a well defined substrate P 1 Ј-binding site in MMPs. Three types of S 1 Ј pockets can be distinguished from the available structures of . One type is a shallow pocket, as found in MMP-1 (human fibroblast collagenase; 13) and 16), where the pockets are limited by the side chains of Arg and Tyr, respectively, crossing the pockets. Many of the structurally known MMPs possess Leu at the corresponding site, and its side chain forms the top of the pocket rather than crossing the pocket. These Leu-containing MMPs may be further classified as deep and intermediate S 1 Ј pocket MMPs. A deep, tunnel-like pocket is found in MMP-3 (stromelysin-1; 12), MMP-12 (metalloelastase; 17), and MMP-14 (MT1-MMP; 21), whereas MMP-2 (gelatinase A; 22), MMP-8 (human neutrophil collagenase; 15), and MMP-9 (gelatinase B; 23) possess an intermedi-

show abstract

Discovery of CGS 27023A, a Non-Peptidic, Potent, and Orally Active Stromelysin Inhibitor That Blocks Cartilage Degradation in Rabbits

Cited by 331 publications

References 24 publications

Stromelysin-1 (MMP-3) is a target and a regulator of Wnt1-induced epithelial-mesenchymal transition (EMT)

Stromelysin-1 (MMP-3) is a target and a regulator of Wnt1-induced epithelial-mesenchymal transition (EMT)

Identification and Characterization of Lutheran Blood Group Glycoprotein as a New Substrate of Membrane-type 1 Matrix Metalloproteinase 1 (MT1-MMP)

The Intermediate S1′ Pocket of the Endometase/Matrilysin-2 Active Site Revealed by Enzyme Inhibition Kinetic Studies, Protein Sequence Analyses, and Homology Modeling

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