, Volume 37, Issue 4, pp 995–1005 | Cite as

Tyrosine Kinase Inhibitor Tyrphostin AG490 Retards Chronic Joint Inflammation in Mice

  • Valeriya Gyurkovska
  • Tsvetanka Stefanova
  • Petya Dimitrova
  • Svetla Danova
  • Rositsa Tropcheva
  • Nina Ivanovska


Tyrphostin AG490 is a Janus kinase (JAK) 2 inhibitor that is clinically used as an anticancer agent and is also effective in various models of inflammatory and autoimmune diseases. In this study, we examined the effects of tyrphostin AG490 on the development of collagenase-induced osteoarthritis (CIOA). Our results showed that tyrphostin-ameliorated cartilage and bone destructions. This effect was associated with decreased expression of signal transducers and activators of transcription 3 (STAT3), phosphorylated JAK2, Dickkopf homolog 1, and receptor activator of nuclear factor κB ligand (RANKL) in the joints of arthritic mice. Tyrphostin AG490 suppressed STAT3 phosphorylation and the expression of tumor necrosis factor-related apoptosis-inducing ligand and RANKL by synovial fluid cells. The drug inhibited RANKL-induced osteoclast differentiation in vitro. Molecules, such as tyrphostin AG490 that limit bone erosion and influence osteoclast generation, might have therapeutic utility in joint degenerative disorders.


collagenase-induced osteoarthritis (CIOA) osteoclasts tyrphostin AG490 STAT3 TRAIL 



This work was supported by a Grant DFNI B01/6 from the National Science Fund, Ministry of Education and Science, Republic of Bulgaria.

Conflict of interest



  1. 1.
    Blom, A.B., P.L. van Lent, A.E. Holthuysen, P.M. van der Kraan, J. Roth, N. van Rooijen, and W.B. van den Berg. 2004. Synovial lining macrophages mediate osteophyte formation during experimental osteoarthritis. Osteoarthritis and Cartilage 12: 627–635.PubMedCrossRefGoogle Scholar
  2. 2.
    Levitzki, A., and E. Mishani. 2006. Tyrphostins and other tyrosine kinase inhibitors. Annual Review of Biochemistry 75: 93–109.PubMedCrossRefGoogle Scholar
  3. 3.
    Meydan, N., T. Grunberger, H. Dadi, M. Shahar, E. Arpaia, Z. Lapidot, J.S. Leeder, M. Freedman, A. Cohen, A. Gazit, A. Levitzki, and C.M. Roifman. 1996. Inhibition of acute lymphoblastic leukaemia by a Jak-2 inhibitor. Nature 379: 645–648.PubMedCrossRefGoogle Scholar
  4. 4.
    Darnell Jr., J.E. 1997. STATs and gene regulation. Science 277: 1630–1635.PubMedCrossRefGoogle Scholar
  5. 5.
    Heim, M.H. 1999. The Jak-STAT pathway: cytokine signalling from the receptor to the nucleus. Journal of Receptor Signal Transduction Research 19: 75–120.PubMedCrossRefGoogle Scholar
  6. 6.
    Igarashi, K., G. Garotta, L. Ozmen, A. Ziemiecki, A.F. Wilks, A.G. Harpur, A.C. Larner, and D.S. Finbloom. 1994. Interferon-gamma induces tyrosine phosphorylation of interferon-gamma receptor and regulated association of protein tyrosine kinases, Jak1 and Jak2, with its receptor. Journal of Biological Chemistry 269: 14333–14336.PubMedGoogle Scholar
  7. 7.
    Davoodi-Semiromi, A., C.H. Wasserfall, C.Q. Xia, R.M. Cooper-DeHoff, M. Wabitsch, M. Clare-Salzler, and M. Atkinson. 2012. The tyrphostin agent tyrphoatin AG490 prevents and reverses type 1 diabetes in NOD mice. PLoS One 7: e36079.PubMedCentralPubMedCrossRefGoogle Scholar
  8. 8.
    Bright, J.J., C. Du, and S. Sriram. 1999. Tyrphostin B42 inhibits IL-12-induced tyrosine phosphorylation and activation of Janus kinase-2 and prevents experimental allergic encephalomyelitis. Journal of Immunology 162: 6255–6262.Google Scholar
  9. 9.
    Constantin, G., S. Brocke, A. Izikson, C. Laudanna, and E.C. Butcher. 1998. Tyrphostin tyrphostin AG490, a tyrosine kinase inhibitor, blocks actively induced experimental autoimmune encephalomyelitis. Euopean Journal of Immunology 28: 3523–3529.CrossRefGoogle Scholar
  10. 10.
    Dimitrova, P., V. Gyurkovska, I. Shalova, L. Saso, and N. Ivanovska. 2009. Inhibition of zymosan-induced kidney dysfunction by tyrphostin AG-490. Journal of Inflammation (Lond) 6: 13.CrossRefGoogle Scholar
  11. 11.
    Pena, G., B. Cai, E.A. Deitch, and L. Ulloa. 2010. JAK2 inhibition prevents innate immune responses and rescues animals from sepsis. Journal of Molecular Medicine (Berlin) 88: 851–859.CrossRefGoogle Scholar
  12. 12.
    Liu, H., Y.M. Yao, Y. Yu, N. Dong, H.N. Yin, and Z.Y. Sheng. 2007. Role of Janus kinase/signal transducer and activator of transcription pathway in regulation of expression and inflammation-promoting activity of high mobility group box protein 1 in rat peritoneal macrophages. Shock 27: 55–60.PubMedCrossRefGoogle Scholar
  13. 13.
    Sareila, O., R. Korhonen, O. Karpanniemi, R. Nieminen, H. Kankaanranta, and E. Moilanen. 2006. JAK inhibitors AG-490 and WHI-P154 decrease IFN-gamma-induced iNOS expression and NO production in macrophages. Mediators of Inflammation 2006: 16161.PubMedCentralPubMedCrossRefGoogle Scholar
  14. 14.
    Nesterov, A., Y. Ivashchenko, and A.S. Kraft. 2002. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) triggers apoptosis in normal prostate epithelial cells. Oncogene 21: 1135–1140.PubMedCrossRefGoogle Scholar
  15. 15.
    Falschlehner, C., U. Schaefer, and H. Walczak. 2009. Following TRAIL’s path in the immune system. Immunology 127: 145–154.PubMedCentralPubMedCrossRefGoogle Scholar
  16. 16.
    Lamhamedi-Cherradi, S.E., S.J. Zheng, K.A. Maguschak, J. Peschon, and Y.H. Chen. 2003. Defective thymocyte apoptosis and accelerated autoimmune diseases in TRAIL-/- mice. Nature Immunology 4: 255–260.PubMedCrossRefGoogle Scholar
  17. 17.
    Song, K., Y. Chen, R. Goke, A. Wilmen, C. Seidel, A. Goke, and B. Hilliard. 2000. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is an inhibitor of autoimmune inflammation and cell cycle progression. Journal of Experimental Medicine 191: 1095–1104.PubMedCentralPubMedCrossRefGoogle Scholar
  18. 18.
    Miranda-Carus, M.E., A. Balsa, M. Benito-Miguel, C.P. De Ayala, and E. Martin-Mola. 2004. Rheumatoid arthritis synovial fluid fibroblasts express TRAIL-R2 (DR5) that is functionally active. Arthritis & Rheumatism 50: 2786–2793.CrossRefGoogle Scholar
  19. 19.
    Weir, E.C., C.W. Lowik, I. Paliwal, and K.L. Insogna. 1996. Colony stimulating factor-1 plays a role in osteoclast formation and function in bone resorption induced by parathyroid hormone and parathyroid hormone-related protein. Journal of Bone and Mineral Research 11: 1474–1481.PubMedCrossRefGoogle Scholar
  20. 20.
    Boyle, W.J., W.S. Simonet, and D.L. Lacey. 2003. Osteoclast differentiation and activation. Nature 423: 337–342.PubMedCrossRefGoogle Scholar
  21. 21.
    Wittrant, Y., S. Theoleyre, S. Couillaud, C. Dunstan, D. Heymann, and F. Redini. 2004. Relevance of an in vitro osteoclastogenesis system to study receptor activator of NF-kB ligand and osteoprotegerin biological activities. Experimental Cell Research 293: 292–301.PubMedCrossRefGoogle Scholar
  22. 22.
    Dimitrova, P., and N. Ivanovska. 2008. Tyrphostin AG-490 inhibited the acute phase of zymosan-induced inflammation. International Immunopharmacology 8: 1567–1577.PubMedCrossRefGoogle Scholar
  23. 23.
    Takahashi, N., N. Udagawa, Y. Kobayashi, and T. Suda. 2007. Generation of osteoclasts in vitro, and assay of osteoclast activity. Methods Molecular Medicine 135: 285–301.CrossRefGoogle Scholar
  24. 24.
    Pelletier, J.P., J. Martel-Pelletier, and S.B. Abramson. 2001. Osteoarthritis, an inflammatory disease: potential implication for the selection of new therapeutic targets. Arthritis & Rheumatism 44: 1237–1247.CrossRefGoogle Scholar
  25. 25.
    Shouda, T., T. Yoshida, T. Hanada, T. Wakioka, M. Oishi, K. Miyoshi, S. Komiya, K. Kosai, Y. Hanakawa, K. Hashimoto, K. Nagata, and A. Yoshimura. 2001. Induction of the cytokine signal regulator SOCS3/CIS3 as a therapeutic strategy for treating inflammatory arthritis. Journal of Clinical Investigations 108: 1781–1788.CrossRefGoogle Scholar
  26. 26.
    Glass 2nd, D.A., P. Bialek, J.D. Ahn, M. Starbuck, M.S. Patel, H. Clevers, M.M. Taketo, F. Long, A.P. McMahon, R.A. Lang, and G. Karsenty. 2005. Canonical Wnt signaling in differentiated osteoblasts controls osteoclast differentiation. Developmental Cell 8: 751–764.PubMedCrossRefGoogle Scholar
  27. 27.
    Krishnan, V., H.U. Bryant, and O.A. Macdougald. 2006. Regulation of bone mass by Wnt signaling. Journal of Clinical Investigations 116: 1202–1209.CrossRefGoogle Scholar
  28. 28.
    Morvan, F., K. Boulukos, P. Clement-Lacroix, S. Roman Roman, I. Suc-Royer, B. Vayssiere, P. Ammann, P. Martin, S. Pinho, P. Pognonec, P. Mollat, C. Niehrs, R. Baron, and G. Rawadi. 2006. Deletion of a single allele of the Dkk1 gene leads to an increase in bone formation and bone mass. Journal of Bone and Mineral Research 21: 934–945.PubMedCrossRefGoogle Scholar
  29. 29.
    Diarra, D., M. Stolina, K. Polzer, J. Zwerina, M.S. Ominsky, D. Dwyer, A. Korb, J. Smolen, M. Hoffmann, C. Scheinecker, D. van der Heide, R. Landewe, D. Lacey, W.G. Richards, and G. Schett. 2007. Dickkopf-1 is a master regulator of joint remodeling. Nature Medicine 13: 156–163.PubMedCrossRefGoogle Scholar
  30. 30.
    Goldring, S.R. and M.B. Goldring. 2004. The role of cytokines in cartilage matrix degeneration in osteoarthritis. Clinical Orthopedics and Related Research (427 Suppl): S27–S36.Google Scholar
  31. 31.
    Livshits, G., G. Zhai, D.J. Hart, B.S. Kato, H. Wang, F.M. Williams, and T.D. Spector. 2009. Interleukin-6 is a significant predictor of radiographic knee osteoarthritis: the Chingford study. Arthritis & Rheumatism 60: 2037–2045.CrossRefGoogle Scholar
  32. 32.
    Choy, E.H., D.A. Isenberg, T. Garrood, S. Farrow, Y. Ioannou, H. Bird, N. Cheung, B. Williams, B. Hazleman, R. Price, K. Yoshizaki, N. Nishimoto, T. Kishimoto, and G.S. Panayi. 2002. Therapeutic benefit of blocking interleukin-6 activity with an anti-interleukin-6 receptor monoclonal antibody in rheumatoid arthritis: a randomized, double-blind, placebo-controlled, dose-escalation TRAIL. Arthritis & Rheumatism 46: 3143–3150.CrossRefGoogle Scholar
  33. 33.
    Yoshizaki, K., N. Nishimoto, M. Mihara, and T. Kishimoto. 1998. Therapy of rheumatoid arthritis by blocking IL-6 signal transduction with a humanized anti-IL-6 receptor antibody. Springer Seminars in Immunopathology 20: 247–259.PubMedCrossRefGoogle Scholar
  34. 34.
    Koenders, M.I., E. Lubberts, B. Oppers-Walgreen, L. van den Bersselaar, M.M. Helsen, F.E. Di Padova, A.M. Boots, H. Gram, L.A. Joosten, and W.B. van den Berg. 2005. Blocking of interleukin-17 during reactivation of experimental arthritis prevents joint inflammation and bone erosion by decreasing RANKL and interleukin-1. American Journal of Pathology 167: 141–149.PubMedCentralPubMedCrossRefGoogle Scholar
  35. 35.
    Van Bezooijen, R.L., L. Van Der Wee-Pals, S.E. Papapoulos, and C.W. Lowik. 2002. Interleukin 17 synergises with tumour necrosis factor alpha to induce cartilage destruction in vitro. Annals of the Rheumatic Diseases 61: 870–876.PubMedCentralPubMedCrossRefGoogle Scholar
  36. 36.
    Catrina, A.I., A.K. Ulfgren, S. Lindblad, L. Grondal, and L. Klareskog. 2002. Low levels of apoptosis and high FLIP expression in early rheumatoid arthritis synovium. Annals of the Rheumatic Diseases 61: 934–936.PubMedCentralPubMedCrossRefGoogle Scholar
  37. 37.
    Jungel, A., C. Ospelt, M. Lesch, M. Thiel, T. Sunyer, O. Schorr, B.A. Michel, R.E. Gay, C. Kolling, C. Flory, S. Gay, and M. Neidhart. 2010. Effect of the oral application of a highly selective MMP-13 inhibitor in three different animal models of rheumatoid arthritis. Annals of the Rheumatic Diseases 69: 898–902.PubMedCentralPubMedCrossRefGoogle Scholar
  38. 38.
    Wu, G., J. Chai, T.L. Suber, J.W. Wu, C. Du, X. Wang, and Y. Shi. 2000. Structural basis of IAP recognition by Smac/DIABLO. Nature 408: 1008–1012.PubMedCrossRefGoogle Scholar
  39. 39.
    Lacey, D.L., E. Timms, H.L. Tan, M.J. Kelley, C.R. Dunstan, T. Burgess, R. Elliott, A. Colombero, G. Elliott, S. Scully, H. Hsu, J. Sullivan, N. Hawkins, E. Davy, C. Capparelli, A. Eli, Y.X. Qian, S. Kaufman, I. Sarosi, V. Shalhoub, G. Senaldi, J. Guo, J. Delaney, and W.J. Boyle. 1998. Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation. Cell 93: 165–176.PubMedCrossRefGoogle Scholar
  40. 40.
    Hsu, H., D.L. Lacey, C.R. Dunstan, I. Solovyev, A. Colombero, E. Timms, H.L. Tan, G. Elliott, M.J. Kelley, I. Sarosi, L. Wang, X.Z. Xia, R. Elliott, L. Chiu, T. Black, S. Scully, C. Capparelli, S. Morony, G. Shimamoto, M.B. Bass, and W.J. Boyle. 1999. Tumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegerin ligand. Proceedings of the National Academy of Sciences U S A 96: 3540–3545.CrossRefGoogle Scholar
  41. 41.
    Ivanovska, N., and P. Dimitrova. 2011. Bone resorption and remodeling in murine collagenase-induced osteoarthritis after administration of glucosamine. Arthritis Research and Therapy 13: R44.PubMedCentralPubMedCrossRefGoogle Scholar
  42. 42.
    Cutolo, M. 2013. The kinase inhibitor tofacitinib in patients with rheumatoid arthritis: latest findings and clinical potential. Therapeutic Advances in Musculoskelet Disease 5: 3–11.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Valeriya Gyurkovska
    • 1
  • Tsvetanka Stefanova
    • 1
  • Petya Dimitrova
    • 1
  • Svetla Danova
    • 2
  • Rositsa Tropcheva
    • 2
  • Nina Ivanovska
    • 1
  1. 1.Department of ImmunologyInstitute of MicrobiologySofiaBulgaria
  2. 2.Section of Microbial GeneticsInstitute of MicrobiologySofiaBulgaria

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