Proteoglycan- and Collagen-Degrading Enzymes from Human Interleukin 1-Stimulated Chondrocytes from Several Species: Proteoglycanase and Collagenase Inhibitors as Potentially New Disease-Modifying Antiarthritic Agents

DiPasquale, G.; Caccese, Robert G.; Pasternak, R D; Conaty, J.; Hubbs, Stephen J.; Perry, K.

doi:10.3181/00379727-183-42416

Cited by 35 publications

(7 citation statements)

References 5 publications

(5 reference statements)

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“…This drug markedly inhibited these MMPs when tested at concentra- tions ranging from 10 -5 to 10 -9 M. Our findings are further supported by studies done by Caputo et al [9] in which rabbit articular cartilage derived proteoglycanase was also blocked in vitro by this drug at concentrations ranging from 10 -6 to 10 -8 M. Our IC 50 value of 1.8 × 10 -9 M for inhibition of MMPproteoglycanases was at least one order of magnitude lower than the previously reported IC 50 values of 6.0 × 10 -8 M [9] and 4.2 × 10 -6 M [16] of the same drug. In addition, studies from DiPasquale et al [16] have shown that U-24522 is approximately 10 times more potent against the proteoglycan-than the collagen-degrading enzymes derived from articular chondrocytes. Current theory suggests that the hydroxamic acid functional group chelates the zinc molecule at the active site of the MMPs thereby inactivating the enzyme [25].…”

Section: Discussioncontrasting

confidence: 73%

“…Hydroxamic acid derivates seem to be the most potent inhibitors of chondrocyte derived proteoglycanases and collagenases compared with carboxyalkyl peptides or thiol peptides [9,16]. We therefore investigated the effect of the hydroxamic acid derivate U-24522 on three aspects of chondrocyte metabolism which were in part modified by the drug: PG synthesis, PG release and activ- Table 3.…”

Section: Discussionmentioning

confidence: 98%

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The proteoglycan metabolism, morphology and viability of articular cartilage treated with a synthetic matrix metalloproteinase inhibitor

1997

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Matrix metalloproteinases (MMP) are among the key enzymes responsible for the proteolytic destruction of articular cartilage during chronic rheumatic diseases. Articular cartilage is one potential target for drugs designed to inhibit the activity of MMPs in order to stop or to slow down the proteolytic destruction of the extracellular matrix of cartilage. The purpose of this study was to investigate the effect of the synthetic inhibitor of MMPs U-24522 for its ability (1) to inhibit in vitro the activity of MMP-proteoglycanases; (2) to modulate the morphology and viability of cartilage explants; and (3) to modify the biosynthesis and release of proteoglycans from articular cartilage explants. U-24522 dose-dependently inhibited the activity of MMP-proteoglycanases and significantly reduced the release of proteoglycans from interleukin-1 treated bovine articular cartilage explants when tested at concentrations ranging from 10(-4) to 10(-9) M. This hydroxamic acid derivative proved not to be harmful to chondrocyte viability and cartilage morphology. In addition, U-24522 had no effect on the rate of proteoglycan biosynthesis of interleukin-1 treated cartilage explants and increased the percentage of newly synthesized proteoglycans to form macromolecular aggregates. Thus U-24522 combines direct inhibitory potential on the activity of MMP-proteoglycanases with the inhibition of interleukin-1 stimulated proteoglycan loss from articular cartilage explants without affecting the morphology, viability and biosynthesis of proteoglycans of bovine articular cartilage explants.

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Section: Discussioncontrasting

confidence: 73%

Section: Discussionmentioning

confidence: 98%

The proteoglycan metabolism, morphology and viability of articular cartilage treated with a synthetic matrix metalloproteinase inhibitor

1997

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“…Preparation of the labeled samples, typically about 1 mM, and of the buffers (containing I O rnM Tris-dll-HCI, pH 7.0, 20 mM CaCI,, 15% acetonitrile43 and 8% D,O) was as described by . The hydroxamatesubstituted inhibitor (IC1 U24522; DiPasquale et al, 1986) was present at a ratio of 1: 1 with the protein for the half-filtered experiments (cf. Ikura .…”

Section: N M R Data Collection and Handlingmentioning

confidence: 99%

“…We present here the solution structure of human stromelysin's catalytic domain under conditions of higher calcium affinity (pH 7.0), and concentration, complexed with a hydroxamate inhibitor of more hydrophobic character ((R,S)-N-[2-[2-(hydroxamino)-2-oxoethyI]-4-methyl-l-oxopentyl]-~-leucyl-~-phenylalaninamide; DiPasquale et al, 1986) and in the presence of acetonitrile as a co-solvent. Our solution structure of the catalytic domain of human stromelysin reveals the binding mode of this peptide substrate analogue in the hydrophobic pockets of the active site (Kinemages 1 and 2) (Protein Data Bank access codes lUMT and IUMS).…”

mentioning

confidence: 99%

Solution structure of the catalytic domain of human stromelysin complexed with a hydrophobic inhibitor

Doren

Kurochkin

et al. 1995

Protein Science

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Stromelysin, a representative matrix metalloproteinase and target of drug development efforts, plays a prominent role in the pathological proteolysis associated with arthritis and secondarily in that of cancer metastasis and invasion. To provide a structural template to aid the development of therapeutic inhibitors, we have determined a medium-resolution structure of a 20-kDa complex of human stromelysin's catalytic domain with a hydrophobic peptidic inhibitor using multinuclear, multidimensional NMR spectroscopy. This domain of this zinc hydrolase contains a mixed 0-sheet comprising one antiparallel strand and four parallel strands, three helices, and a methionine-containing turn near the catalytic center. The ensemble of 20 structures was calculated using, on average, 8 interresidue NOE restraints per residue for the 166-residue protein fragment complexed with a 4-residue substrate analogue. The mean RMS deviation (RMSD) to the average structure for backbone heavy atoms is 0.91 A and for all heavy atoms is 1.42 A. The structure has good stereochemical properties, including its backbone torsion angles. The &sheet and a-helices of the catalytic domains of human stromelysin (NMR model) and human fibroblast collagenase (X-ray crystallographic model of Lovejoy B et al., 1994b, Biochemisfry 33:8207-8217) superimpose well, having a pairwise RMSD for backbone heavy atoms of 2.28 A when three loop segments are disregarded. The hydroxamate-substituted inhibitor binds across the hydrophobic active site of stromelysin in an extended conformation. The first hydrophobic side chain is deeply buried in the principal Si subsite, the second hydrophobic side chain is located on the opposite side of the inhibitor backbone in the hydrophobic Si surface subsite, and a third hydrophobic side chain (Pi) lies at the surface.Keywords: catalytic domain; hydroxamate; matrix metalloproteinase 3; multidimensional NMR; protein structure; stromelysinAs a member of the zinc-and calcium-dependent family of matrix metalloproteinases (MMPs), which hydrolyze the extracellular matrix, stromelysin participates in the tissue remodeling of health and disease. Regarding the shared domain structure of the MMPs, their subfamilies, specificity, transcriptional regulation, activation, and posttranslational regulation, see reviews of Woessner (1991), Matrisian (1992), Birkedal-Hansen et al. (1993), andNagase (1995). The involvement of MMPs in tissue remodeling includes that of development, tissue resorption, reproduction, angiogenesis, and wound healing, and that of sev-

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“…Most interestingly, the interaction of cells (cell products) associated with the inflammatory lesion (monocytes and T-cells) with synovial cells or chondrocytes can induce collagenase synthesis in these cells [12][13][14]. Interleukin-1 (IL-1), a macrophage or monocyte product, has been identified as a potent stimulator of collagenase synthesis [15][16][17][18][19]. Other agents such as T-cell products [14], uric acid crystals [20] and phorbol esters [21,22] have also been shown to stimulate connective tissue degrading activity by synovial cells, chondrocytes or cartilage.…”

Section: Introductionmentioning

confidence: 99%

The effect of calcium channel blockers and calmodulin inhibitors on the macrophage factor-stimulated synthesis of collagenase by rabbit chondrocytes

Nolan¹,

Gathright²,

Wagner³

1988

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Macrophages and monocytes secrete a factor(s) which can stimulate the synthesis of collagenase in synovial cells and in chondrocytes. Incubation of rabbit chondrocytes with macrophage conditioned medium (MCM) and with the calcium channel blockers, nifedipine, verapamil or diltiazem (up to 200 microM) had no effect on collagenase synthesis. However, TMB-8 (8-[N,N-diethylamino]-octyl 3,4,5-trimethoxybenzoate hydrochloride), an inhibitor of internal calcium movement, did inhibit the process with an IC50 of approximately 130 microM. The calmodulin antagonists, trifluoperazine, chlorpromazine and calmidazolium (R-24571) were effective inhibitors of the process with IC50's of 40 microM, 18 microM and 3.5 microM, respectively. Collagenase activity itself was not affected by these agents. The data suggests that calmodulin and/or internal calcium movement may play a role in the macrophage factor-stimulated synthesis of collagenase in rabbit chondrocytes.

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Proteoglycan- and Collagen-Degrading Enzymes from Human Interleukin 1-Stimulated Chondrocytes from Several Species: Proteoglycanase and Collagenase Inhibitors as Potentially New Disease-Modifying Antiarthritic Agents

Cited by 35 publications

References 5 publications

The proteoglycan metabolism, morphology and viability of articular cartilage treated with a synthetic matrix metalloproteinase inhibitor

The proteoglycan metabolism, morphology and viability of articular cartilage treated with a synthetic matrix metalloproteinase inhibitor

Solution structure of the catalytic domain of human stromelysin complexed with a hydrophobic inhibitor

The effect of calcium channel blockers and calmodulin inhibitors on the macrophage factor-stimulated synthesis of collagenase by rabbit chondrocytes

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