Aggrecanase 1

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Yasunori Okada - One of the best experts on this subject based on the ideXlab platform.

  • development of human neutralizing antibody to adamts4 Aggrecanase 1 and adamts5 Aggrecanase 2
    Biochemical and Biophysical Research Communications, 2016
    Co-Authors: Aya Shiraishi, Kanehisa Kojoh, Akira Miyakoshi, Satsuki Mochizuki, Yasunori Okada
    Abstract:

    ADAMTS4 (Aggrecanase-1) and ADAMTS5 (Aggrecanase-2), members of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) gene family, are considered to play a key role in aggrecan degradation of articular cartilage in human osteoarthritis. Here, we developed a neutralizing antibody to these Aggrecanases by screening human combinatorial antibody library. Among the five candidate antibodies, one antibody was immunoreactive with both ADAMTS4 and ADAMTS5, showing no or negligible cross-reactivity with 10 different related metalloproteinases of the ADAMTS, ADAM (a disintegrin and metalloproteinase) and MMP (matrix metalloproteinase) gene families. This antibody almost completely and partially inhibited Aggrecanase activity of ADAMTS4 and ADAMTS5, respectively. It also suppressed the Aggrecanase activity derived from interleukin-1-stimulated osteoarthritic chondrocytes. These data demonstrate that the antibody is specific to ADAMTS4 and ADAMTS5 and inhibits their Aggrecanase activity at molecular and cellular levels, and suggest that this antibody may be useful for treatment of pathological conditions such as osteoarthritis.

  • ccn1 cyr61 is overexpressed in human osteoarthritic cartilage and inhibits adamts 4 Aggrecanase 1 activity
    Arthritis & Rheumatism, 2015
    Co-Authors: Miyuki Chijiiwa, Tokuhiro Kimura, Yutaka Fujii, Yoshiaki Toyama, Satsuki Mochizuki, Yukie Tanaka, Hidenori Shimizu, Hiroyuki Enomoto, Yasunori Okada
    Abstract:

    Objective ADAMTS-4, also called Aggrecanase 1, is considered to play a key role in aggrecan degradation in human osteoarthritic (OA) cartilage, but information about regulators of ADAMTS-4 Aggrecanase activity remains limited. We undertook this study to search for molecules that modulate ADAMTS-4 activity. Methods Molecules copurified with ADAMTS-4 from ADAMTS-4–transfected chondrocytic cells were sequenced by nanoscale liquid chromatography tandem mass spectrometry. Binding activity was determined by immunoprecipitation and solid-phase binding assay. Effects on ADAMTS-4 activity were examined by aggrecan digestion assay. Expression of the binding molecule in OA cartilage and chondrocytes was examined by immunohistochemistry and reverse transcription–polymerase chain reaction. Results We identified CCN1 (Cyr61) as an ADAMTS-4–binding protein and showed specific binding to the ADAMTS-4 cysteine-rich domain. Aggrecanase activity of ADAMTS-4 was inhibited by interaction with CCN1. Expression of messenger RNA for CCN1 was significantly higher in human OA cartilage than in normal cartilage. CCN1 was immunolocalized to chondrocytes in OA cartilage, showing direct correlations of immunoreactivity with the Mankin score of cartilage lesions and chondrocyte cloning. CCN1 and ADAMTS-4 were commonly coexpressed in clustered chondrocytes. CCN1 expression in OA chondrocytes was down-regulated by interleukin-1α (IL-1α) and up-regulated by transforming growth factor β (TGFβ). ADAMTS-4 expression was induced by treatment with IL-1α or TGFβ, but Aggrecanase activity was detected only under stimulation with IL-1α. TGFβ-treated chondrocytes exhibited Aggrecanase activity when CCN1 expression was knocked down. Conclusion Our findings provide the first evidence that CCN1 suppresses ADAMTS-4 activity and that CCN1 overexpression is directly correlated with chondrocyte cloning in OA cartilage. Our results suggest that the TGFβ/CCN1 axis plays a role in chondrocyte cluster formation through inhibition of ADAMTS-4.

  • CCN1 (Cyr61) Is Overexpressed in Human Osteoarthritic Cartilage and Inhibits ADAMTS‐4 (Aggrecanase 1) Activity
    Arthritis & Rheumatism, 2015
    Co-Authors: Miyuki Chijiiwa, Tokuhiro Kimura, Yutaka Fujii, Yoshiaki Toyama, Satsuki Mochizuki, Yukie Tanaka, Hidenori Shimizu, Hiroyuki Enomoto, Yasunori Okada
    Abstract:

    Objective ADAMTS-4, also called Aggrecanase 1, is considered to play a key role in aggrecan degradation in human osteoarthritic (OA) cartilage, but information about regulators of ADAMTS-4 Aggrecanase activity remains limited. We undertook this study to search for molecules that modulate ADAMTS-4 activity. Methods Molecules copurified with ADAMTS-4 from ADAMTS-4–transfected chondrocytic cells were sequenced by nanoscale liquid chromatography tandem mass spectrometry. Binding activity was determined by immunoprecipitation and solid-phase binding assay. Effects on ADAMTS-4 activity were examined by aggrecan digestion assay. Expression of the binding molecule in OA cartilage and chondrocytes was examined by immunohistochemistry and reverse transcription–polymerase chain reaction. Results We identified CCN1 (Cyr61) as an ADAMTS-4–binding protein and showed specific binding to the ADAMTS-4 cysteine-rich domain. Aggrecanase activity of ADAMTS-4 was inhibited by interaction with CCN1. Expression of messenger RNA for CCN1 was significantly higher in human OA cartilage than in normal cartilage. CCN1 was immunolocalized to chondrocytes in OA cartilage, showing direct correlations of immunoreactivity with the Mankin score of cartilage lesions and chondrocyte cloning. CCN1 and ADAMTS-4 were commonly coexpressed in clustered chondrocytes. CCN1 expression in OA chondrocytes was down-regulated by interleukin-1α (IL-1α) and up-regulated by transforming growth factor β (TGFβ). ADAMTS-4 expression was induced by treatment with IL-1α or TGFβ, but Aggrecanase activity was detected only under stimulation with IL-1α. TGFβ-treated chondrocytes exhibited Aggrecanase activity when CCN1 expression was knocked down. Conclusion Our findings provide the first evidence that CCN1 suppresses ADAMTS-4 activity and that CCN1 overexpression is directly correlated with chondrocyte cloning in OA cartilage. Our results suggest that the TGFβ/CCN1 axis plays a role in chondrocyte cluster formation through inhibition of ADAMTS-4.

  • hyaluronan inhibits expression of adamts4 Aggrecanase 1 in human osteoarthritic chondrocytes
    Annals of the Rheumatic Diseases, 2009
    Co-Authors: Taku Yatabe, Masayuki Takizawa, Aiko Okada, Miyuki Chijiiwa, Tokuhiro Kimura, Yoshinari Fujita, Yoshiaki Toyama, Hideo Matsumoto, Satsuki Mochizuki, Yasunori Okada
    Abstract:

    Background: Intra-articular injection of hyaluronan (HA) has been suggested to have a disease-modifying effect in osteoarthritis, but little is known about the possible mechanisms. Objective: To investigate the effects of HA species of different molecular mass, including 800 kDa (HA800) and 2700 kDa (HA2700), on the expression of Aggrecanases (ie, ADAMTS species), which play a key role in aggrecan degradation. Methods: The effects of HA species on the expression of ADAMTS1, 4, 5, 8, 9 and 15 in interleukin 1α (IL1α)-stimulated osteoarthritic chondrocytes were studied by reverse transcription PCR and real-time PCR. Expression of ADAMTS4 protein and Aggrecanase activity and signal transduction pathways of IL1, CD44 and intracellular adhesion molecule 1 (ICAM1) were examined by immunoblotting. Results: IL1α treatment of chondrocytes induced ADAMTS4, and HA800 and HA2700 significantly decreased IL1α-induced expression of ADAMTS4 mRNA and protein. IL1α-stimulated Aggrecanase activity in osteoarthritic chondrocytes was reduced by treatment with HA2700 or transfection of small interfering RNA for ADAMTS4. A similar result was obtained when HA2700 was added to explant cultures of osteoarthritic cartilage. HA2700 neither directly inhibited nor bound to ADAMTS4. Downregulation of ADAMTS4 expression by HA2700 was attenuated by treatment of IL1α-treated chondrocytes with antibodies to CD44 and/or ICAM1. The increased phosphorylation of IL1 receptor-associated kinase-1 and extracellular signal-regulated protein kinase1/2 induced by the IL1α treatment was downregulated by enhanced IRAK-M expression after HA2700 treatment. Conclusion: These data suggest that HA2700 suppresses aggrecan degradation by downregulating IL1α-induced ADAMTS4 expression through the CD44 and ICAM1 signalling pathways in osteoarthritic chondrocytes.

  • calcium pentosan polysulfate directly inhibits enzymatic activity of adamts4 Aggrecanase 1 in osteoarthritic chondrocytes
    FEBS Letters, 2008
    Co-Authors: Masayuki Takizawa, Taku Yatabe, Aiko Okada, Miyuki Chijiiwa, Satsuki Mochizuki, Peter Ghosh, Yasunori Okada
    Abstract:

    Aggrecanases that include ADAMTS1, 4, 5, 8, 9 and 15 are considered to play key roles in aggrecan degradation in osteoarthritic cartilage. Here we demonstrate that calcium pentosan polysulfate (CaPPS) directly inhibits the Aggrecanase activity of ADAMTS4 without affecting the mRNA expression of the ADAMTS species in interleukin-1α-stimulated osteoarthritic chondrocytes. Synthetic peptides corresponding to specific regions of the thrombospondin type 1 repeat, cysteine-rich or spacer domain of ADAMTS4 inhibit the binding to immobilized CaPPS. These data suggest that CaPPS could function as chondroprotective agent for the treatment of osteoarthritis by inhibition of ADAMTS4 through interaction with the C-terminal ancillary domain.

Elizabeth C. Arner - One of the best experts on this subject based on the ideXlab platform.

  • substrate dependent inhibition kinetics of an active site directed inhibitor of adamts 4 Aggrecanase 1
    Biochemistry, 2007
    Co-Authors: Arthur J Wittwer, Elizabeth C. Arner, Anne-marie Malfait, Robert L Hills, Grace E Munie, R H Keith, C P Anglin, Micky D. Tortorella
    Abstract:

    ADAMTS-4 (Aggrecanase-1) is implicated in the breakdown of articular cartilage and is an attractive target for therapeutic intervention in arthritis. Cleavage of the native substrate, aggrecan, occurs through exosite interactions and peptide sequence recognition. Although expected to be competitive with aggrecan, the hydroxamic acid, SC81956, demonstrated noncompetitive inhibition kinetics with a K i of 23 nM. The IC 50 of SC81956 did not change when aggrecan was varied from 12.8 to 200 nM (0.2-3.3 times the apparent aggrecan K m of 61 nM) but was shifted as expected for a competitive inhibitor when increasing levels of a low molecular weight peptide substrate were added to a fluorogenic peptide assay system. These observations are consistent with a model for aggrecan cleavage where substrate initially binds at an exosite, followed by binding of the appropriate peptide sequence at the active site. A peptide-competitive inhibitor could bind both free enzyme and initial substrate-enzyme exosite complex but would be excluded by the final Michaelis complex. Noncompetitive appearing kinetics for such inhibitors is predicted as long as the equilibrium between the two forms of enzyme-substrate complex significantly favors the initial exosite complex. In support, hydrolysis of a low molecular weight peptide substrate and its inhibition by SC81956 were unaffected by aggrecan concentrations substantially above the K m . These observations suggest that the apparent K m for aggrecan cleavage predominately reflects the exosite interaction. Consequently, the efficacy of active-site inhibitors of ADAMTS-4 will not be limited by competition with native substrate as predicted from the K m determined by traditional kinetic models.

  • adamts 4 Aggrecanase 1 n terminal activation mechanisms
    Archives of Biochemistry and Biophysics, 2005
    Co-Authors: Micky D. Tortorella, Elizabeth C. Arner, Robert L Hills, Jennifer A Gormley, Lyle E Pegg, Grace E Munie, Anne-marie Malfait
    Abstract:

    Abstract ADAMTS-4 (Aggrecanase 1) is synthesized as a latent precursor protein that may require activation through removal of its prodomain before it can exert catalytic activity. We examined various proteinases as well as auto-activation under a wide range of conditions for removal of the prodomain and induction of enzymatic activity. The proprotein convertases, furin, PACE4, and PC5/6 efficiently removed the prodomain through cleavage at Arg 212 /Phe 213 , generating an active enzyme. Of a broad range of proteases evaluated, only MMP-9 and trypsin were capable of removing the prodomain. In the presence of mercuric compounds, removal of the prodomain through autocatalysis was not observed, nor was it observed at temperatures from 22 to 65 °C, at ionic strengths from 0.1 to 1 M, or at acidic/neutral pH. At basic pH 8–10, removal of the prodomain by autocatalysis occurred, generating an active enzyme. In conclusion, the pro-form of ADAMTS-4 is not catalytically active and only a limited number of mechanisms mediate its N-terminal activation.

  • proprotein convertase furin interacts with and cleaves pro adamts4 Aggrecanase 1 in the trans golgi network
    Journal of Biological Chemistry, 2004
    Co-Authors: Ping Wang, Kristen England, Micky D. Tortorella, Anne-marie Malfait, Gary Thomas, Elizabeth C. Arner
    Abstract:

    Abstract A member of the A disintegrin and metalloproteinase domain with thrombospondin type-1 motifs (ADAMTS-4) protease family can efficiently cleave aggrecan at several sites detected in joints of osteoarthritic patients. Although recent studies have shown that removal of the prodomain of ADAMTS4 is critical for its ability to degrade aggrecan, the cellular mechanisms for its processing and trafficking remain unclear. In this study, by using both furin-specific inhibitor and RNA interference technique, we demonstrate that furin plays an important role in the intracellular removal of ADAMTS4 prodomain. Further, we demonstrate that proADAMTS4 can be processed by means of multiple furin recognition sites: 206RPRR209, 209RAKR212, or 211KR212. The processing of proADAMTS4 was completely blocked by brefeldin A treatment, suggesting that processing occurs in the trans-Golgi network. Indeed, ADAMTS4 is co-localized with furin in trans-Golgi network. Interestingly, the pro form of ADAMTS4, not its mature one, co-precipitates with furin, suggesting that furin physically interacts with the prodomain of ADAMTS-4. In addition, our evidence suggests that a furin-independent pathway may also contribute to the activation of ADAMTS4. These results indicate that the activation mechanism for ADAMTS4 can be targeted for therapeutical intervention against this enzyme.

  • Characterization of human Aggrecanase 2 (ADAM-TS5): substrate specificity studies and comparison with Aggrecanase 1 (ADAM-TS4).
    Matrix Biology, 2002
    Co-Authors: Micky D. Tortorella, Timothy Burn, Robert C Newton, Elizabeth C. Arner
    Abstract:

    ADAM-TS5 (Aggrecanase 2), one of two cartilage Aggrecanases is a member of the ADAM protein family. Like ADAM-TS4 (Aggrecanase 1) the enzyme cleaves cartilage aggrecan at the Glu373–Ala374 bond, a marker of Aggrecanase activity. In this study we have characterized the substrate specificity of ADAM-TS5 and compared it with that of ADAM-TS4. The recombinant human ADAM-TS5, like ADAM-TS4 cleaves aggrecan at Glu1480–Gly1481, Glu1667–Gly1668, Glu1771–Ala1772 and Glu1871–Leu1872 bonds more readily than at the Glu373–Ala374 bond. In addition, ADAM-TS5 exhibited an additional site of cleavage in the region spanning residues Gly1481 and Glu1667, representing a unique cleavage of ADAM-TS5. ADAM-TS5 cleaved aggrecan approximately 2-fold slower than ADAM-TS4. Neither ADAM-TS5 nor ADAM-TS4 was able to cleave the extracellular matrix proteins fibronectin, thrombospondin, type I collagen, type II collagen, gelatin or general protein substrates such as casein and transferrin. Finally, the zymogen of stromelysin (MMP-3) was not activated by either ADAM-TS4 or ADAM-TS5.

  • expression and regulation of Aggrecanase in arthritis the role of tgf β
    Journal of Immunology, 2002
    Co-Authors: Yuji Yamanishi, Micky D. Tortorella, Elizabeth C. Arner, Melody Clark, David L Boyle, Rich A Maki, Gary S Firestein
    Abstract:

    Aggrecanases are key matrix-degrading enzymes that act by cleaving aggrecan at the Glu373-Ala374 site. While these fragments have been detected in osteoarthritis (OA) and rheumatoid arthritis (RA) cartilage and synovial fluid, no information is available on the regulation or expression of the two key Aggrecanases (Aggrecanase-1 and Aggrecanase-2) in synovial tissue (ST) or fibroblast-like synoviocytes (FLS). The Aggrecanase-1 gene was constitutively expressed by both RA and OA FLS. Real-time PCR demonstrated that TGF-β significantly increased Aggrecanase-1 gene expression in FLS. Aggrecanase-1 induction peaked after 24 h of TGF-β stimulation. The expression of Aggrecanase-1 mRNA was significantly greater in RA ST than in OA or nonarthritis ST. Aggrecanase-2 mRNA and protein were constitutively produced by nonarthritis, OA, and RA FLS but were not increased by IL-1, TNF-α, or TGF-β. Furthermore, OA, RA, and nonarthritis ST contained similar amounts of immunoreactive Aggrecanase-2. The major form of the Aggrecanase-2 enzyme was 70 kDa in nonarthritis ST, whereas a processed 53-kDa form was abundant in RA ST. Therefore, Aggrecanase-1 and -2 are differentially regulated in FLS. Both are constitutively expressed, but Aggrecanase-1 is induced by cytokines, especially TGF-β. In contrast, Aggrecanase-2 protein may be regulated by a post-translational mechanism in OA and RA ST. Synovial and FLS production of Aggrecanase can contribute to cartilage degradation in RA and OA.

Micky D. Tortorella - One of the best experts on this subject based on the ideXlab platform.

  • substrate dependent inhibition kinetics of an active site directed inhibitor of adamts 4 Aggrecanase 1
    Biochemistry, 2007
    Co-Authors: Arthur J Wittwer, Elizabeth C. Arner, Anne-marie Malfait, Robert L Hills, Grace E Munie, R H Keith, C P Anglin, Micky D. Tortorella
    Abstract:

    ADAMTS-4 (Aggrecanase-1) is implicated in the breakdown of articular cartilage and is an attractive target for therapeutic intervention in arthritis. Cleavage of the native substrate, aggrecan, occurs through exosite interactions and peptide sequence recognition. Although expected to be competitive with aggrecan, the hydroxamic acid, SC81956, demonstrated noncompetitive inhibition kinetics with a K i of 23 nM. The IC 50 of SC81956 did not change when aggrecan was varied from 12.8 to 200 nM (0.2-3.3 times the apparent aggrecan K m of 61 nM) but was shifted as expected for a competitive inhibitor when increasing levels of a low molecular weight peptide substrate were added to a fluorogenic peptide assay system. These observations are consistent with a model for aggrecan cleavage where substrate initially binds at an exosite, followed by binding of the appropriate peptide sequence at the active site. A peptide-competitive inhibitor could bind both free enzyme and initial substrate-enzyme exosite complex but would be excluded by the final Michaelis complex. Noncompetitive appearing kinetics for such inhibitors is predicted as long as the equilibrium between the two forms of enzyme-substrate complex significantly favors the initial exosite complex. In support, hydrolysis of a low molecular weight peptide substrate and its inhibition by SC81956 were unaffected by aggrecan concentrations substantially above the K m . These observations suggest that the apparent K m for aggrecan cleavage predominately reflects the exosite interaction. Consequently, the efficacy of active-site inhibitors of ADAMTS-4 will not be limited by competition with native substrate as predicted from the K m determined by traditional kinetic models.

  • adamts 4 Aggrecanase 1 n terminal activation mechanisms
    Archives of Biochemistry and Biophysics, 2005
    Co-Authors: Micky D. Tortorella, Elizabeth C. Arner, Robert L Hills, Jennifer A Gormley, Lyle E Pegg, Grace E Munie, Anne-marie Malfait
    Abstract:

    Abstract ADAMTS-4 (Aggrecanase 1) is synthesized as a latent precursor protein that may require activation through removal of its prodomain before it can exert catalytic activity. We examined various proteinases as well as auto-activation under a wide range of conditions for removal of the prodomain and induction of enzymatic activity. The proprotein convertases, furin, PACE4, and PC5/6 efficiently removed the prodomain through cleavage at Arg 212 /Phe 213 , generating an active enzyme. Of a broad range of proteases evaluated, only MMP-9 and trypsin were capable of removing the prodomain. In the presence of mercuric compounds, removal of the prodomain through autocatalysis was not observed, nor was it observed at temperatures from 22 to 65 °C, at ionic strengths from 0.1 to 1 M, or at acidic/neutral pH. At basic pH 8–10, removal of the prodomain by autocatalysis occurred, generating an active enzyme. In conclusion, the pro-form of ADAMTS-4 is not catalytically active and only a limited number of mechanisms mediate its N-terminal activation.

  • proprotein convertase furin interacts with and cleaves pro adamts4 Aggrecanase 1 in the trans golgi network
    Journal of Biological Chemistry, 2004
    Co-Authors: Ping Wang, Kristen England, Micky D. Tortorella, Anne-marie Malfait, Gary Thomas, Elizabeth C. Arner
    Abstract:

    Abstract A member of the A disintegrin and metalloproteinase domain with thrombospondin type-1 motifs (ADAMTS-4) protease family can efficiently cleave aggrecan at several sites detected in joints of osteoarthritic patients. Although recent studies have shown that removal of the prodomain of ADAMTS4 is critical for its ability to degrade aggrecan, the cellular mechanisms for its processing and trafficking remain unclear. In this study, by using both furin-specific inhibitor and RNA interference technique, we demonstrate that furin plays an important role in the intracellular removal of ADAMTS4 prodomain. Further, we demonstrate that proADAMTS4 can be processed by means of multiple furin recognition sites: 206RPRR209, 209RAKR212, or 211KR212. The processing of proADAMTS4 was completely blocked by brefeldin A treatment, suggesting that processing occurs in the trans-Golgi network. Indeed, ADAMTS4 is co-localized with furin in trans-Golgi network. Interestingly, the pro form of ADAMTS4, not its mature one, co-precipitates with furin, suggesting that furin physically interacts with the prodomain of ADAMTS-4. In addition, our evidence suggests that a furin-independent pathway may also contribute to the activation of ADAMTS4. These results indicate that the activation mechanism for ADAMTS4 can be targeted for therapeutical intervention against this enzyme.

  • Characterization of human Aggrecanase 2 (ADAM-TS5): substrate specificity studies and comparison with Aggrecanase 1 (ADAM-TS4).
    Matrix Biology, 2002
    Co-Authors: Micky D. Tortorella, Timothy Burn, Robert C Newton, Elizabeth C. Arner
    Abstract:

    ADAM-TS5 (Aggrecanase 2), one of two cartilage Aggrecanases is a member of the ADAM protein family. Like ADAM-TS4 (Aggrecanase 1) the enzyme cleaves cartilage aggrecan at the Glu373–Ala374 bond, a marker of Aggrecanase activity. In this study we have characterized the substrate specificity of ADAM-TS5 and compared it with that of ADAM-TS4. The recombinant human ADAM-TS5, like ADAM-TS4 cleaves aggrecan at Glu1480–Gly1481, Glu1667–Gly1668, Glu1771–Ala1772 and Glu1871–Leu1872 bonds more readily than at the Glu373–Ala374 bond. In addition, ADAM-TS5 exhibited an additional site of cleavage in the region spanning residues Gly1481 and Glu1667, representing a unique cleavage of ADAM-TS5. ADAM-TS5 cleaved aggrecan approximately 2-fold slower than ADAM-TS4. Neither ADAM-TS5 nor ADAM-TS4 was able to cleave the extracellular matrix proteins fibronectin, thrombospondin, type I collagen, type II collagen, gelatin or general protein substrates such as casein and transferrin. Finally, the zymogen of stromelysin (MMP-3) was not activated by either ADAM-TS4 or ADAM-TS5.

  • expression and regulation of Aggrecanase in arthritis the role of tgf β
    Journal of Immunology, 2002
    Co-Authors: Yuji Yamanishi, Micky D. Tortorella, Elizabeth C. Arner, Melody Clark, David L Boyle, Rich A Maki, Gary S Firestein
    Abstract:

    Aggrecanases are key matrix-degrading enzymes that act by cleaving aggrecan at the Glu373-Ala374 site. While these fragments have been detected in osteoarthritis (OA) and rheumatoid arthritis (RA) cartilage and synovial fluid, no information is available on the regulation or expression of the two key Aggrecanases (Aggrecanase-1 and Aggrecanase-2) in synovial tissue (ST) or fibroblast-like synoviocytes (FLS). The Aggrecanase-1 gene was constitutively expressed by both RA and OA FLS. Real-time PCR demonstrated that TGF-β significantly increased Aggrecanase-1 gene expression in FLS. Aggrecanase-1 induction peaked after 24 h of TGF-β stimulation. The expression of Aggrecanase-1 mRNA was significantly greater in RA ST than in OA or nonarthritis ST. Aggrecanase-2 mRNA and protein were constitutively produced by nonarthritis, OA, and RA FLS but were not increased by IL-1, TNF-α, or TGF-β. Furthermore, OA, RA, and nonarthritis ST contained similar amounts of immunoreactive Aggrecanase-2. The major form of the Aggrecanase-2 enzyme was 70 kDa in nonarthritis ST, whereas a processed 53-kDa form was abundant in RA ST. Therefore, Aggrecanase-1 and -2 are differentially regulated in FLS. Both are constitutively expressed, but Aggrecanase-1 is induced by cytokines, especially TGF-β. In contrast, Aggrecanase-2 protein may be regulated by a post-translational mechanism in OA and RA ST. Synovial and FLS production of Aggrecanase can contribute to cartilage degradation in RA and OA.

Katy E Georgiadis - One of the best experts on this subject based on the ideXlab platform.

  • SP0019 AGG-523: Aggrecanase-selective inhibitor for the treatment of osteoarthritis
    Annals of the Rheumatic Diseases, 2013
    Co-Authors: Katy E Georgiadis
    Abstract:

    Osteoarthritis (OA) is the most common form of arthritis affecting nearly 27 million people in the United States. OA is caused by the loss of articular cartilage due to increased production of proteolytic enzymes such as matrix metalloproteinases (MMPs) and Aggrecanases. Depletion of aggrecan is one of the earliest changes observed in osteoarthritis. Aggrecan is a chondroitin sulfate and keratan sulfate-bearing proteoglycan. Two cartilage Aggrecanases, Aggrecanase 1 (ADAMTS4) and Aggrecanase 2 (ADAMTS5) cleave aggrecan within the interglobular domain between residues Glu373 and Ala 374. There are no disease-modifying therapies for the treatment of OA. The current study describes the chondroprotective effect of an oral Aggrecanase-selective inhibitor (AGG-523) in a rat model of osteoarthritis and demonstrates that the significant protection, and lack of phenotypic alteration, observed with gene-deleted ADAMTS-5 and ADAMTS-4/5 mice, can also be observed with the use of a selective ADAMTS-4/5 inhibitor in the rat meniscal tear model of OA. This paradigm of selective ADAMTS-4/5 inhibition avoids the side-effects observed clinically and pre-clinically with broad-spectrum MMP inhibition. Disclosure of Interest K. Georgiadis Shareholder of: Pfizer, Employee of: Pfizer

  • synthesis and biological evaluation of 4 keto phenoxy methyl biphenyl 4 sulfonamides a class of potent Aggrecanase 1 inhibitors
    Bioorganic & Medicinal Chemistry Letters, 2009
    Co-Authors: Darrin William Hopper, Jason Shaoyun Xiang, Yonghan Hu, Jennifer R Thomason, Joshua James Sabatini, Manus Ipek, Matthew D Vera, Eric Feyfant, Qin Wang, Katy E Georgiadis
    Abstract:

    Abstract The prevention of aggrecan (a key component of cartilage) cleavage via the inhibition of Aggrecanase-1 may provide a unique opportunity to stop the progression of cartilage degradation in osteoarthritis. The evaluation of a series of biphenylsulfonamides resulted in the identification of the ((4-keto)-phenoxy)methyl biphenyl-4-sulfonamides analogs (19–21 and 24) with improved Agg-1 inhibition and MMP-2, MMP-13 activity.

  • n 8 hydroxy 5 substituted quinolin 7 yl phenyl methyl 2 phenyloxy amino acetamide inhibitors of adamts 5 Aggrecanase 2
    Bioorganic & Medicinal Chemistry Letters, 2008
    Co-Authors: Adam M Gilbert, Erica Reifenberg, Katy E Georgiadis, Carl R Flannery, Matthew Gregory Bursavich, Sabrina Lombardi, Elisabeth A Morris
    Abstract:

    Abstract N-((8-Hydroxy-5-substituted-quinolin-7-yl)(phenyl)methyl)-2-phenyloxy/amino-acetamide inhibitors of ADAMTS-5 (Aggrecanase-2) have been prepared. Selected compounds 10, 14, 25, and 53 show sub-μM ADAMTS-5 potency and good selectivity over the related metalloproteases ADAMTS-4 (Aggrecanase-1), MMP-13, and MMP-12. Compound 53 shows a good balance of potent ADAMTS-5 inhibition, moderate CYP3A4 inhibition and good rat liver microsome stability. This series of compounds represents progress towards selective ADAMTS-5 inhibitors as disease modifying osteoarthritis agents.

  • N-((8-hydroxy-5-substituted-quinolin-7-yl)(phenyl)methyl)-2-phenyloxy/amino-acetamide inhibitors of ADAMTS-5 (Aggrecanase-2).
    Bioorganic & medicinal chemistry letters, 2008
    Co-Authors: Adam M Gilbert, Erica Reifenberg, Katy E Georgiadis, Carl R Flannery, Matthew Gregory Bursavich, Sabrina Lombardi, Elisabeth A Morris
    Abstract:

    Abstract N-((8-Hydroxy-5-substituted-quinolin-7-yl)(phenyl)methyl)-2-phenyloxy/amino-acetamide inhibitors of ADAMTS-5 (Aggrecanase-2) have been prepared. Selected compounds 10, 14, 25, and 53 show sub-μM ADAMTS-5 potency and good selectivity over the related metalloproteases ADAMTS-4 (Aggrecanase-1), MMP-13, and MMP-12. Compound 53 shows a good balance of potent ADAMTS-5 inhibition, moderate CYP3A4 inhibition and good rat liver microsome stability. This series of compounds represents progress towards selective ADAMTS-5 inhibitors as disease modifying osteoarthritis agents.

  • 5 phenyl 3 h spiro indoline 3 2 1 3 4 thiadiazol 2 one inhibitors of adamts 5 Aggrecanase 2
    Bioorganic & Medicinal Chemistry Letters, 2007
    Co-Authors: Matthew Gregory Bursavich, Erica Reifenberg, Katy E Georgiadis, Carl R Flannery, Adam M Gilbert, Sabrina Lombardi, Elisabeth A Morris
    Abstract:

    Abstract 5′-Phenyl-3′ H -spiro[indoline-3,2′-[1,3,4]thiadiazol]-2-one inhibitors of ADAMTS-5 (Aggrecanase-2) have been prepared via commercially available starting materials. Selected compounds 23 , 33 – 35 show sub-micromolar ADAMTS-5 potency and strong SAR trends with selectivity over the related metalloproteases ADAMTS-4 (Aggrecanase-1), MMP12, and MMP13. This series of compounds represents progress toward a selective ADAMTS-5 inhibitor as a disease modifying osteoarthritis drug.

Elisabeth A Morris - One of the best experts on this subject based on the ideXlab platform.

  • n 8 hydroxy 5 substituted quinolin 7 yl phenyl methyl 2 phenyloxy amino acetamide inhibitors of adamts 5 Aggrecanase 2
    Bioorganic & Medicinal Chemistry Letters, 2008
    Co-Authors: Adam M Gilbert, Erica Reifenberg, Katy E Georgiadis, Carl R Flannery, Matthew Gregory Bursavich, Sabrina Lombardi, Elisabeth A Morris
    Abstract:

    Abstract N-((8-Hydroxy-5-substituted-quinolin-7-yl)(phenyl)methyl)-2-phenyloxy/amino-acetamide inhibitors of ADAMTS-5 (Aggrecanase-2) have been prepared. Selected compounds 10, 14, 25, and 53 show sub-μM ADAMTS-5 potency and good selectivity over the related metalloproteases ADAMTS-4 (Aggrecanase-1), MMP-13, and MMP-12. Compound 53 shows a good balance of potent ADAMTS-5 inhibition, moderate CYP3A4 inhibition and good rat liver microsome stability. This series of compounds represents progress towards selective ADAMTS-5 inhibitors as disease modifying osteoarthritis agents.

  • N-((8-hydroxy-5-substituted-quinolin-7-yl)(phenyl)methyl)-2-phenyloxy/amino-acetamide inhibitors of ADAMTS-5 (Aggrecanase-2).
    Bioorganic & medicinal chemistry letters, 2008
    Co-Authors: Adam M Gilbert, Erica Reifenberg, Katy E Georgiadis, Carl R Flannery, Matthew Gregory Bursavich, Sabrina Lombardi, Elisabeth A Morris
    Abstract:

    Abstract N-((8-Hydroxy-5-substituted-quinolin-7-yl)(phenyl)methyl)-2-phenyloxy/amino-acetamide inhibitors of ADAMTS-5 (Aggrecanase-2) have been prepared. Selected compounds 10, 14, 25, and 53 show sub-μM ADAMTS-5 potency and good selectivity over the related metalloproteases ADAMTS-4 (Aggrecanase-1), MMP-13, and MMP-12. Compound 53 shows a good balance of potent ADAMTS-5 inhibition, moderate CYP3A4 inhibition and good rat liver microsome stability. This series of compounds represents progress towards selective ADAMTS-5 inhibitors as disease modifying osteoarthritis agents.

  • 5 phenyl 3 h spiro indoline 3 2 1 3 4 thiadiazol 2 one inhibitors of adamts 5 Aggrecanase 2
    Bioorganic & Medicinal Chemistry Letters, 2007
    Co-Authors: Matthew Gregory Bursavich, Erica Reifenberg, Katy E Georgiadis, Carl R Flannery, Adam M Gilbert, Sabrina Lombardi, Elisabeth A Morris
    Abstract:

    Abstract 5′-Phenyl-3′ H -spiro[indoline-3,2′-[1,3,4]thiadiazol]-2-one inhibitors of ADAMTS-5 (Aggrecanase-2) have been prepared via commercially available starting materials. Selected compounds 23 , 33 – 35 show sub-micromolar ADAMTS-5 potency and strong SAR trends with selectivity over the related metalloproteases ADAMTS-4 (Aggrecanase-1), MMP12, and MMP13. This series of compounds represents progress toward a selective ADAMTS-5 inhibitor as a disease modifying osteoarthritis drug.

  • 5-((1H-pyrazol-4-yl)methylene)-2-thioxothiazolidin-4-one inhibitors of ADAMTS-5.
    Bioorganic & medicinal chemistry letters, 2006
    Co-Authors: Adam M Gilbert, Erica Reifenberg, Katy E Georgiadis, Carl R Flannery, Matthew Gregory Bursavich, Sabrina Lombardi, Elisabeth A Morris
    Abstract:

    A series of 5-((1H-pyrazol-4-yl)methylene)-2-thioxothiazolidin-4-one inhibitors of ADAMTS-5 (Aggrecanase-2) is described. These compounds show microM functional inhibition of ADAMTS-5, and represent a new class of agents with the potential of inhibiting degradation of aggrecan, a major component of cartilage which is lost in osteoporosis. Compound 12 is noteworthy in that it has an ADAMTS-5 IC50: 1.1 microM and shows >40-fold functional selectivity over ADAMTS-4 (Aggrecanase-1).

  • release of hyaluronan and hyaladherins aggrecan g1 domain and link proteins from articular cartilage exposed to adamts 4 Aggrecanase 1 or adamts 5 Aggrecanase 2
    Arthritis & Rheumatism, 2004
    Co-Authors: Priya S Chockalingam, Weilan Zeng, Elisabeth A Morris, Carl R Flannery
    Abstract:

    Objective To determine whether Aggrecanase (ADAMTS) activities in articular cartilage can directly lead to the release of hyaluronan (HA) and hyaladherins (aggrecan G1 domain and link proteins), as may occur ex vivo during stimulation of cartilage explants with interleukin-1 (IL-1) or retinoic acid or in vivo in synovial joints during aging and joint pathology. Methods Bovine articular cartilage discs (live or freeze-killed) were cultured in the presence of IL-1 or were incubated in digestion buffer containing recombinant human ADAMTS-4 (rHuADAMTS-4; Aggrecanase 1) or rHuADAMTS-5 (Aggrecanase 2). Culture media, digestion supernatants, and tissue extracts were assayed for sulfated glycosaminoglycan (sGAG) content and analyzed by Western blotting to detect Aggrecanase-generated G1 domain (using neoepitope monoclonal antibody AGG-C1/anti-NITEGE373) and link proteins (using monoclonal antibody 8-A-4), as well as by quantitative enzyme-linked immunosorbent assays to detect Aggrecanase-generated G1 domain (G1-NITEGE373) and HA. Results IL-1 treatment of live cartilage explants induced a time-dependent release of sGAG, Aggrecanase-generated G1 domain (G1-NITEGE373), and HA into the culture media. Exposure of live or freeze-killed articular cartilage discs to rHuADAMTS-4 or rHuADAMTS-5 resulted in a dose- and time-dependent release of sGAG and hyaluronan from the tissue, accompanied by a concomitant release of functionally intact hyaladherins (aggrecan G1-NITEGE373 and link proteins). Conclusion Coincident with aggrecanolysis, Aggrecanase activities in articular cartilage may actuate the release of HA and associated hyaladherins, thereby further compromising the integrity of the cartilage matrix during degenerative joint diseases such as osteoarthritis.