Download PDF
Planta Med 2012; 78(10): 951-956
DOI: 10.1055/s-0031-1298625
Biological and Pharmacological Activity
Original Papers
Georg Thieme Verlag KG Stuttgart · New York

Library-based Discovery of DYRK1A/CLK1 Inhibitors from Natural Product Extracts

Patrick Grabher
1   Division of Pharmaceutical Biology, University of Basel, Basel, Switzerland
,
Emilie Durieu
2   Protein Phosphorylation & Human Disease Group, Station Biologique de Roscoff, CNRS, Roscoff, France
,
Eirini Kouloura
3   Laboratory of Pharmacognosy and Natural Products Chemistry, School of Pharmacy, University of Athens, Athens, Greece
,
Maria Halabalaki
3   Laboratory of Pharmacognosy and Natural Products Chemistry, School of Pharmacy, University of Athens, Athens, Greece
,
Leandros A. Skaltsounis
3   Laboratory of Pharmacognosy and Natural Products Chemistry, School of Pharmacy, University of Athens, Athens, Greece
,
Laurent Meijer
2   Protein Phosphorylation & Human Disease Group, Station Biologique de Roscoff, CNRS, Roscoff, France
4   ManRos Therapeutics, Centre de Perharidy, Roscoff, France
,
Matthias Hamburger
1   Division of Pharmaceutical Biology, University of Basel, Basel, Switzerland
,
Olivier Potterat
1   Division of Pharmaceutical Biology, University of Basel, Basel, Switzerland
› Author Affiliations
Further Information

Publication History

received 02 March 2012
revised 07 May 2012

accepted 08 May 2012

Publication Date:
06 June 2012 (online)

    Permissions and Reprints

Abstract

The dual specificity tyrosine-phosphorylation-regulated kinase DYRK1A possesses diverse roles in neuronal development and adult brain physiology, and increased activity has been linked to neurodegenerative diseases. Very few inhibitors of this kinase have been reported up to now. Screening of a library of > 900 plant and fungal extracts afforded 25 extracts with IC50s < 10 µg/mL against DYRK1A. To identify the active constituents, the extracts were submitted to a process integrating physicochemical data with biological information, referred to as HPLC-based activity profiling. Follow-up investigation of four extracts led to the targeted isolation of harmine (1, IC50 0.022 µM) from Peganum harmala, emodin (3, IC50 4.2 µM) from Cassia nigricans, kaempferol (4, IC50 0.91 µM) from Cuscuta chinensis, and 3,8-di-O-methylherbacetin (11, IC50 8.6 µM), 3,3′,4′-tri-O-methylmyricetin (12, IC50 7.1 µM) and ombuin (15, IC50 1.7 µM) from Larrea tridentata as the active constituents. Active extracts and compounds were also tested on the closely related cdc2-like kinase CLK1. Finally, the selectivity profile of compounds was evaluated by including other members of the DYRKs and CLKs families. While the flavonoids and emodin did not show significant differences in the potency of their activities, harmine (1) was most active against DYRK1A, CLK1, and CLK4, and less potent against the other kinases, with selectivity ranging from 2- to 20-fold.

Key words

library-based natural product screening - DYRK1A - CLK1 - HPLC-based activity profiling - harmine - emodin - flavonoids - Peganum harmala - Zygophyllaceae - Cuscuta chinensis - Convolvulaceae - Larrea tridentate - Zygophyllaceae - Cassia nigricans - Caesalpiniaceae

Supporting Information

 

  • References

  • 1 Wegiel J, Gong CX, Hwang YW. The role of DYRK1A in neurodegenerative diseases. FEBS J 2011; 278: 236-245
    CrossrefPubMedGoogle Scholar
  • 2 Koo KA, Kim ND, Chon YS, Jung MS, Lee BJ, Kim JH, Song WJ. QSAR analysis of pyrazolidine-3,5-diones derivatives as Dyrk1A inhibitors. Bioorg Med Chem Lett 2009; 19: 2321-2328
    PubMedGoogle Scholar
  • 3 Rosenthal AS, Tanega C, Shen M, Mott BT, Bougie JM, Nguyen DT, Mistelli T, Auld DS, Maloney DJ, Thomas CJ. Potent and selective small molecule inhibitors of specific isoforms of Cdc2-like kinases (Clk) and dual specificity tyrosine-phosphorylation-regulated kinases (Dyrk). Bioorg Med Chem Lett 2011; 21: 3152-3158
    CrossrefPubMedGoogle Scholar
  • 4 Bain J, Plater L, Elliott M, Shpiro N, Hastie CJ, McLauchlan H, Klevernic I, Arthur JSC, Alessi DR, Cohen P. The selectivity of protein kinase inhibitors: a further update. Biochem J 2007; 408: 297-315
    CrossrefPubMedGoogle Scholar
  • 5 Adayev T, Chen-Hwang MC, Murakami N, Wegiel J, Hwang YW. Kinetic properties of a MNB/DYRK1A mutant suitable for the elucidation of biochemical pathways. Biochemistry 2006; 45: 12011-12019
    CrossrefPubMedGoogle Scholar
  • 6 Guedj F, Sébrié C, Rivals I, Ledru A, Paly E, Bizot JC, Smith D, Rubin E, Gillet B, Arbones M, Delabar JM. Green tea polyphenols rescue of brain defects induced by overexpression of DYRK1A. PLoS One 2009; 4: e4606
    CrossrefPubMedGoogle Scholar
  • 7 Ogawa Y, Nonaka Y, Goto T, Ohnishi E, Hiramatsu T, Kii I, Yoshida M, Ikura T, Onogi H, Shibuya H, Hosoya T, Ito N, Hagiwara M. Development of a novel selective inhibitor of the Down syndrome-related kinase Dyrk1A. Nat Commun 2010; 1: 86
    PubMedGoogle Scholar
  • 8 Fedorov O, Huber K, Eisenreich A, Filippakopoulos P, King O, Bullock AN, Szklarczyk D, Jensen LJ, Fabbro D, Trappe J, Rauch U, Bracher F, Knapp S. Specific CLK inhibitors from a novel chemotype for regulation of alternative splicing. Chem Biol 2011; 18: 67-76
    CrossrefPubMedGoogle Scholar
  • 9 Debdab M, Carreaux F, Renault S, Soundararajan M, Fedorov O, Filippakopoulos P, Lozach O, Babault L, Tatouh T, Baratte B, Ogawa Y, Hagiwara M, Eisenreich A, Rauch U, Knapp S, Meijer L, Bazureau JP. Leucettines, a class of potent inhibitors of cdc2-like kinases and dual specificity, tyrosine phosphorylation regulated kinases derived from the marine sponge leucettamine B. Modulation of alternative pre-RNA splicing. J Med Chem 2011; 54: 4172-4186
    CrossrefPubMedGoogle Scholar
  • 10 Hagiwara M. Alternative splicing: a new drug target of the post-genome era. Biochim Biophys Acta 2005; 1754: 324-331
    CrossrefPubMedGoogle Scholar
  • 11 Markham KR. Techniques of flavonoid identification. London: Academic Press; 1982: 39
    Google Scholar
  • 12 Potterat O, Hamburger M. Natural products in drug discovery – Concepts and approaches for tracking bioactivity. Curr Org Chem 2006; 10: 899-920
    CrossrefPubMedGoogle Scholar
  • 13 Joshi BS, Bai Y, Puar MS, Dubose KK, Pelletier SW. 1H- and 13C-NMR assignments for some pyrrolo[2,1b]-quinazoline alkaloids of Adhatoda vasica . J Nat Prod 1994; 57: 953-962
    CrossrefPubMedGoogle Scholar
  • 14 Ye M, Yan Y, Qiao L, Ni X. Studies on chemical constituents of Cuscuta chinensis . China J Chin Mater Med 2002; 27: 115-117
    PubMedGoogle Scholar
  • 15 Jung MJ, Chung HY, Choi JH, Choi JS. Antioxidant principles from the needles of red pine Pinus densiflora . Phytother Res 2003; 17: 1064-1068
    CrossrefPubMedGoogle Scholar
  • 16 Qiu SX, Lu ZZ, Luyengi L, Lee SK, Pezzuto JM, Farnsworth NR, Thompson LU, Fong HHS. Isolation and characterization of flaxseed (Linum usitatissimum) constituents. Pharm Biol 1999; 37: 1-7
    CrossrefPubMedGoogle Scholar
  • 17 Gedara S, Abdel-Halim OB, El-Sharkawy SH, Salama OM, Shier TW, Halim AF. New erythroxane type diterpenoids from Fagonia boveana (Hadidi) Hadidi & Graf. Z Naturforsch 1995; 58: 23-32
    PubMedGoogle Scholar
  • 18 Sarmento da Silva TM, Geraldo de Carvalho M, Braz-Filho R. Estudo espectroscopico em elucidaçao estructural de flavonoides de Solanum jabrense Agra & Nee e S. paludosum Moric. Quim Nova 2009; 32: 1119-1128
    PubMedGoogle Scholar
  • 19 Wollenweber E, Fischer R, Doerr M, Irvine K, Pereira C, Stevens JF. Chemodiversity of exudate flavonoids in Cassinia and Ozothamnus (Asteraceae, Gnaphalieae). Z Naturforsch C 2008; 63: 731-739
    PubMedGoogle Scholar
  • 20 Chavez MH, Roque NF, Ayres MCC. Steroids and flavonoids of Porcelia macrocarpa . J Braz Chem Soc 2004; 15: 608-613
    CrossrefPubMedGoogle Scholar
  • 21 Bernhard H, Thiele K. Additional flavonoids from the leaves of Larrea tridentata . Planta Med 1981; 41: 100-103
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 Sakakibara M, Di Feo Jr. D, Nakatani N, Timmermann B, Mabry TJ. Flavonoid methyl ethers on the external leaf surface of Larrea tridentata and L. divaricata . Phytochemistry 1976; 15: 727-731
    CrossrefPubMedGoogle Scholar
  • 23 Sakakibara M, Timmermann BN, Nakatani N, Waldrum H, Mabry TJ. New 8-hydroxyflavonols from Larrea tridentata . Phytochemistry 1975; 14: 849-851
    CrossrefPubMedGoogle Scholar
  • 24 Abou-Gazar H, Bedir E, Takamatsu S, Ferreira D, Khan IA. Antioxidant lignans from Larrea tridentata . Phytochemistry 2004; 65: 2499-2505
    CrossrefPubMedGoogle Scholar
  • 25 Adayev T, Wegiel J, Hwang YW. Harmine is an ATP-competitive inhibitor for dual-specificity tyrosine phosphorylation-regulated kinase 1A (Dyrk1A). Arch Biochem Biophys 2011; 507: 212-218
    CrossrefPubMedGoogle Scholar
  • 26 Frost D, Meechoovet B, Wang T, Gately S, Giorgetti M, Shcherbakova I, Dunckley T. β-Carboline compounds, including harmine, inhibit DYRK1A and tau phosphorylation at multiple Alzheimerʼs disease-related sites. PLoS One 2011; 6: e19264
    CrossrefPubMedGoogle Scholar
  • 27 Göckler N, Jofre G, Papadopoulos C, Soppa U, Tejedor FJ, Becker W. Harmine specifically inhibits protein kinase DYRK1A and interferes with neurite formation. FEBS J 2009; 2276: 6324-6337
    PubMedGoogle Scholar
  • 28 Seifert A, Allan LA, Clarke PR. DYRK1A phosphorylates caspase 9 at an inhibitory site and is potently inhibited in human cells by harmine. FEBS J 2008; 275: 6268-6280
    CrossrefPubMedGoogle Scholar
  • 29 Ahmadu A, Abdulkarim A, Grougnet R, Myrianthopoulos V, Tillequin F, Magiatis P, Skaltsounis AL. Two new peltogynoids from Acacia nilotica Delile with kinase inhibitory activity. Planta Med 2010; 76: 458-460
    Article in Thieme ConnectPubMedGoogle Scholar