New Insights from a Focused Library Approach Aiming at Development of Inhibitors of Dual-Specificity Protein Phosphatases

  • Go Hirai
  • Ayako Tsuchiya
  • Mikiko Sodeoka
Conference paper


Protein phosphorylations are important biological reactions participating in intracellular signal transductions [1], and they control a diverse set of cellular events including cell proliferation, cell-cycle regulation, and differentiation. Key players in protein phosphorylations are protein kinases and protein phosphatases, which dynamically control the levels of the phosphorylated proteins in cells. Phosphorylation mainly occurs at the hydroxyl groups of serine, threonine, and tyrosine residues in eukaryotic cells [2]. Protein kinases and phosphatases are classified according to their substrate specificity of phosphorylation and dephosphorylation. Human genome analysis indicates that there are 518 protein kinases [3], which can be divided into 90 tyrosine kinases and 428 serine/threonine kinases [4]. Dysfunction of protein kinases is well established to be implicated in the disruption of normal signal transduction pathways and in tumor formation. Therefore, kinase inhibitors are expected to be useful as biological tools and therapeutic agents, so there is both academic and commercial interest in developing specific inhibitors for certain protein kinases [5]. A number of such inhibitors, such as Gleevec, Iressa, Tarceva, and Sutent, have been approved as therapeutic agents by the Food and Drug Administration in the United States.


Protein Phosphatase Core Structure Potent Inhibitory Activity Hydrophobic Alkyl Chain Enzyme Selectivity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was supported in part by a grant from the Uehara Memorial Foundation (Japan) and project funding from RIKEN (Japan). We would like to thank Prof. Hiroyuki Osada, Prof. Siro Simizu, Prof. Takeo Usui, Dr. Keisuke Ishida, and Dr. Ngit Shin Lai (RIKEN, Japan) for their collaboration in this work. We are also grateful to our co-workers at RIKEN, Ms. Kana Oonuma, Mr. Yusuke Koyama, Dr. Yuko Otani, and other coworkers and collaborators listed in the references, for their contributions.


  1. 1.
    Hunter T (2000) Signaling-2000 and beyond. Cell 100:113–127CrossRefPubMedGoogle Scholar
  2. 2.
    Olsen JV, Blagoev B, Gnad F, Macek B, Kumar C, Mortensen P, Mann M (2006) Global, in vivo, and site-specific phosphorylation dynamics in signaling networks. Cell 127:635–648CrossRefPubMedGoogle Scholar
  3. 3.
    Manning G, Whyte DB, Martinez R, Hunter T, Sudarsanam S (2002) The protein kinase complement of the human genome. Science 298:1912–1934CrossRefPubMedGoogle Scholar
  4. 4.
    Johnson SA, Hunter T (2005) Kinomics: methods for deciphering the kinome. Nat Methods 2:17–25CrossRefPubMedGoogle Scholar
  5. 5.
    Grant SK (2009) Therapeutic protein kinase inhibitors. Cell Mol Life Sci 66:1163–1177CrossRefPubMedGoogle Scholar
  6. 6.
    Alonso A, Sasin J, Bottini N, Friedberg I, Osterman A, Godzik A, Hunter T, Dixon J, Mustelin T (2004) Protein tyrosine phosphatases in the human genome. Cell 117:699–711CrossRefPubMedGoogle Scholar
  7. 7.
    Shi Y (2009) Serine/threonine phosphatases: mechanism through structure. Cell 139:468–484CrossRefPubMedGoogle Scholar
  8. 8.
    Patterson KI, Brummer T, O’Brien PM, Daly RJ (2009) Dual-specificity phosphatases: critical regulators with diverse cellular targets. Biochem J 418:475–489PubMedGoogle Scholar
  9. 9.
    Rudolph J (2007) Cdc25 phosphatases: structure, specificity, and mechanism. Biochemistry 46:3595–3604CrossRefPubMedGoogle Scholar
  10. 10.
    Boutros R, Dozier C, Ducommun B (2006) The when and wheres of CDC25 phosphatases. Curr Opin Cell Biol 18:185–191CrossRefPubMedGoogle Scholar
  11. 11.
    Boutros T, Chevet E, Metrakos P (2008) Mitogen-activated protein (MAP) kinase/MAP kinase phosphatase regulation: roles in cell growth, death, and cancer. Pharmacol Rev 60:261–310CrossRefPubMedGoogle Scholar
  12. 12.
    Malumbres M, Barbacid M (2009) Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer 9:153–166CrossRefPubMedGoogle Scholar
  13. 13.
    Hamaguchi T, Sudo T, Osada H (1995) RK-682, a potent inhibitor of tyrosine phosphatase, arrested the mammalian cell cycle progression at G1phase. FEBS Lett 372:54–58CrossRefPubMedGoogle Scholar
  14. 14.
    Cerignoli F, Rahmouni S, Ronai Z, Mustelin T (2006) Regulation of MAP kinases by the VHR dual-specific phosphatase: implications for cell growth and differentiation. Cell Cycle 5:2210–2215CrossRefPubMedGoogle Scholar
  15. 15.
    Shinagawa S, Muroi M, Ito T, Hida T (1993) Preparation of tetronic acid derivatives as phospholipase A2 inhibitors. Application: JP Patent 1991-190103, 05043568Google Scholar
  16. 16.
    Sodeoka M, Sampe R, Kagamizono T, Osada H (1996) Asymmetric synthesis of RK-682 and its analogs, and evaluation of their protein phosphatase inhibitory activities. Tetrahedron Lett 37:8775–8778CrossRefGoogle Scholar
  17. 17.
    Ishibashi T, Bottaro DP, Chan A, Miki T, Aaronson SA (1992) Expression cloning of a human dual-specificity phosphatase. Proc Natl Acad Sci USA 89:12170–12174CrossRefPubMedGoogle Scholar
  18. 18.
    Todd JL, Tanner KG, Denu JM (1999) Extracellular regulated kinases (ERK) 1 and ERK2 are authentic substrates for the dual-specificity protein-tyrosine phosphatase VHR. A novel role in down-regulating the ERK pathway. J Biol Chem 274:13271–13280CrossRefPubMedGoogle Scholar
  19. 19.
    Alonso A, Saxena M, Williams S, Mustelin T (2001) Inhibitory role for dual specificity phosphatase VHR in T cell antigen receptor and CD28-induced Erk and Jnk activation. J Biol Chem 276:4766–4771CrossRefPubMedGoogle Scholar
  20. 20.
    Miller JL (2006) Recent developments in focused library design: targeting gene-families. Curr Top Med Chem 6:19–29CrossRefPubMedGoogle Scholar
  21. 21.
    Stockwell BR (2004) Exploring biology with small organic molecules. Nature (Lond) 432:846–854CrossRefGoogle Scholar
  22. 22.
    Yuvaniyama J, Denu JM, Dixon JE, Saper MA (1996) Crystal structure of the dual specificity protein phosphatase VHR. Science 272:1328–1331CrossRefPubMedGoogle Scholar
  23. 23.
    Fauman EB, Cogswell JP, Lovejoy B, Rocque WJ, Holmes W, Montana VG, Piwnica-Worms H, Rink MJ, Saper MA (1998) Crystal structure of the catalytic domain of the human cell cycle control phosphatase, Cdc25A. Cell 93:617–625CrossRefPubMedGoogle Scholar
  24. 24.
    Sodeoka M, Sampe R, Kojima S, Baba Y, Usui T, Ueda K, Osada H (2001) Synthesis of a tetronic acid library focused on inhibitors of tyrosine and dual-specificity protein phosphatases and its evaluation regarding VHR and cdc25B inhibition. J Med Chem 44:3216–3222CrossRefPubMedGoogle Scholar
  25. 25.
    Usui T, Kojima S, Kidokoro S, Ueda K, Osada H, Sodeoka M (2001) Design and synthesis of a dimeric derivative of RK-682 with increased inhibitory activity against VHR, a dual-specificity ERK phosphatase: implications for the molecular mechanism of the inhibition. Chem Biol 8:1209–1220CrossRefPubMedGoogle Scholar
  26. 26.
    Sodeoka M, Baba Y (2001) Development of protein phosphatases inhibitors: a focused library approach. J Synth Org Chem Jpn 59:1095–1102CrossRefGoogle Scholar
  27. 27.
    Ishida K, Teruya T, Simizu S, Osada H (2004) Exploitation of heparanase inhibitors from microbial metabolites using an efficient visual screening system. J Antibiot (Tokyo) 57:136–142CrossRefGoogle Scholar
  28. 28.
    Ishida K, Hirai G, Murakami K, Teruya T, Simizu S, Sodeoka M, Osada H (2004) Structure-based design of a selective heparanase inhibitor as an antimetastatic agent. Mol Cancer Ther 3:1069–1077CrossRefPubMedGoogle Scholar
  29. 29.
    Saito K, Yamagichu T (1978) NMR spectroscopic studies of the tautomerism in tetramic acid analogs and their anilides. III. Polar solvent effects on the tautomeric populations. Bull Chem Soc Jpn 51:651–652CrossRefGoogle Scholar
  30. 30.
    Hirai G, Tsuchiya A, Koyama Y, Otani Y, Oonuma K, Dodo K, Simizu S, Osada H, Sodeoka M (2011) Development of a Vaccinia H1-related (VHR) phosphatase inhibitor with a nonacidic phosphate-mimicking core structure. ChemMedChem 6:617–622CrossRefPubMedGoogle Scholar
  31. 31.
    Rahmouni S, Cerignoli F, Alonso A, Tsutji T, Henkens R, Zhu C, Louis-dit-Sully C, Moutschen M, Jiang W, Mustelin T (2006) Loss of the VHR dual-specific phosphatase causes cell-cycle arrest and senescence. Nat Cell Biol 8:524–531CrossRefPubMedGoogle Scholar
  32. 32.
    Kang TH, Kim KT (2006) Negative regulation of ERK activity by VRK3-mediated activation of VHR phosphatase. Nat Cell Biol 8:863–869CrossRefPubMedGoogle Scholar
  33. 33.
    Henkens R, Delvenne P, Arafa M, Moutschen M, Zeddou M, Tautz L, Boniver J, Mustelin T, Rahmouni S (2008) Cervix carcinoma is associated with an up-regulation and nuclear localization of the dual-specificity protein phosphatase VHR. BMC Cancer 8:147–155CrossRefPubMedGoogle Scholar
  34. 34.
    Arnoldussen YJ, Lorenzo PI, Pretorius ME, Waehre H, Risberg B, Maelandsmo GM, Danielsen HE, Saatcioglu F (2008) The mitogen-activated protein kinase phosphatase vaccinia H1-related protein inhibits apoptosis in prostate cancer cells and is overexpressed in prostate cancer. Cancer Res 68:9255–9264CrossRefPubMedGoogle Scholar
  35. 35.
    Hamaguchi T, Masuda A, Morino T, Osada H (1997) Stevastelins, a novel group of immunosuppressants, inhibit dual-specificity protein phosphatases. Chem Biol 4:279–286CrossRefPubMedGoogle Scholar
  36. 36.
    Cebula RE, Blanchard JL, Boisclair MD, Pal K, Bockovich NJ (1997) Synthesis and phosphatase inhibitory activity of analogs of sulfircin. Bioorg Med Chem Lett 7:2015–2020CrossRefGoogle Scholar
  37. 37.
    Bergnes G, Gilliam CL, Boisclair MD, Blanchard JL, Blake KV, Epstein DM, Pal K (1999) Generation of an Ugi library of phosphate mimic-containing compounds and identification of novel dual specific phosphatase inhibitors. Bioorg Med Chem Lett 9:2849–2854CrossRefPubMedGoogle Scholar
  38. 38.
    Ueda K, Usui T, Nakayama H, Ueki M, Takio K, Ubukata M, Osada H (2002) 4-Isoavenaciolide covalently binds and inhibits VHR, a dual-specificity phosphatase. FEBS Lett 525:48–52CrossRefPubMedGoogle Scholar
  39. 39.
    Wu S, Vossius S, Rahmouni S, Miletic AV, Vang T, Vazquez-Rodriguez J, Cerignoli F, Arimura Y, Williams S, Hayes T, Moutschen M, Vasile S, Pellecchia M, Mustelin T, Tautz L (2009) Multidentate small-molecule inhibitors of vaccinia H1-related (VHR) phosphatase decrease proliferation of cervix cancer cells. J Med Chem 52:6716–6723CrossRefPubMedGoogle Scholar
  40. 40.
    Wang Z, Zhang B, Wang M, Carr BI (2005) Cdc25A and ERK interaction: EGFR-independent ERK activation by a protein phosphatase Cdc25A inhibitor, compound 5. J Cell Physiol 204:437–444CrossRefPubMedGoogle Scholar
  41. 41.
    Xia K, Lee RS, Narsimhan RP, Mukhopadhyay NK, Neel BG, Roberts TM (1999) Tyrosine phosphorylation of the proto-oncoprotein Raf-1 is regulated by Raf-1 itself and the phosphatase Cdc25A. Mol Cell Biol 19:4819–4824PubMedGoogle Scholar

Copyright information

© Springer 2012

Authors and Affiliations

  1. 1.Synthetic Organic Chemistry Laboratory, RIKEN Advanced Science InstituteWako, SaitamaJapan

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