Cancer Chemotherapy and Pharmacology

, Volume 59, Issue 2, pp 157–164 | Cite as

Inhibitors of kinesin Eg5: antiproliferative activity of monastrol analogues against human glioblastoma cells

  • Christine Müller
  • Dietmar Gross
  • Vasiliki Sarli
  • Michael Gartner
  • Athanassios Giannis
  • Günther Bernhardt
  • Armin Buschauer
Original Article


The inhibition of kinesin Eg5 by small molecules such as monastrol is currently evaluated as an approach to develop a novel class of antiproliferative drugs for the treatment of malignant tumours. Therefore, we studied the effects of the new monastrol analogues enastron, dimethylenastron and vasastrol VS-83 on the proliferation of human glioblastoma cells in the kinetic crystal violet assay. Compared to monastrol, the new cell cycle specific compounds showed an at least one order of magnitude higher anti proliferative activity against U-87 MG, U-118 MG, and U-373 MG glioblastoma cells. The compounds were neither inactivated by hydrolysis nor by binding to serum proteins. Moreover, we demonstrated the characteristic monoaster formation after incubation of cells with the new compounds by confocal laser scanning microscopy. We also showed that the arrangement of β-actin and tubulin, vital components of the cyto-skeleton of mitotic and quiescent cells, were not affected by the new compounds. Due to the necessity of overcoming the blood–brain barrier in the treatment of brain tumours, we investigated if the new monastrol analogues are modulators or substrates of the p-glycoprotein (p-gp) 170 by a flow cytometric calcein-AM efflux assay. The tested compounds showed no modulating effects on the p-gp function. With respect to the treatment of primary and secondary CNS tumours, the results of our experiments suggest that the new monastrol analogues represent an interesting class of potential anticancer drugs, predicted to be less neurotoxic in comparison to classical tubulin inhibitors.


Kinesin inhibitors Monastrol analogues Human glioblastoma cells Chemosensitivity Confocal microscopy 



This work was supported by Grant MRTN CT-2004-512348 (Spindle Dynamics) from the European Commission. We also thank the Fonds der Chemischen Industrie for financial support.


  1. 1.
    Wood KW, Cornwell WD, Jackson JR (2001) Past and future of the mitotic spindle as an oncology target. Curr Opin Pharmacol 1:370–377PubMedCrossRefGoogle Scholar
  2. 2.
    Blangy A, Lane HA, d´Herin P, Harper M, Kress M, Nigg EA (1995) Phosphorylation by p34cdc2 regulates spindle association of human Eg5, a kinesin-related motor essential for bipolar spindle formation in vivo. Cell 83:1159–1169PubMedCrossRefGoogle Scholar
  3. 3.
    Quasthoff S, Hartung HP (2002) Chemotherapy-induced peripheral neuropathy. J Neurol 249:9–17PubMedCrossRefGoogle Scholar
  4. 4.
    Mayer TU, Kapoor TM, Haggarty SJ, King RW, Schreiber SL, Mitchison TJ (1999) Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen. Science 286:971–974PubMedCrossRefGoogle Scholar
  5. 5.
    Gartner M, Sunder-Plassmann N, Seiler J, Utz M, Vernos I, Surrey T, Giannis A (2005) Development and biological evaluation of potent and specific inhibitors of mitotic Kinesin Eg5. Chembiochem 6:1173–1177PubMedCrossRefGoogle Scholar
  6. 6.
    Maliga Z, Kapoor TM, Mitchison TJ (2002) Evidence that monastrol is an allosteric inhibitor of the mitotic kinesin Eg5. Chem Biol 9:989–996PubMedCrossRefGoogle Scholar
  7. 7.
    Sarli V, Huemmer S, Sunder-Plassmann N, Mayer TU, Giannis A (2005) Synthesis and biological evaluation of novel Eg5 inhibitors. Chembiochem 6:2005–2013PubMedCrossRefGoogle Scholar
  8. 8.
    Bernhardt G, Reile H, Birnbock H, Spruss T, Schonenberger H (1992) Standardized kinetic microassay to quantify differential chemosensitivity on the basis of proliferative activity. J Cancer Res Clin Oncol 118:35–43PubMedCrossRefGoogle Scholar
  9. 9.
    Brier S, Lemaire D, Debonis S, Forest E, Kozielski F (2004) Identification of the protein binding region of S-trityl-l-cysteine, a new potent inhibitor of the mitotic kinesin Eg5. Biochemistry 43:13072–13082PubMedCrossRefGoogle Scholar
  10. 10.
    Hollo Z, Homolya L, Davis CW, Sarkadi B (1994) Calcein accumulation as a fluorometric functional assay of the multidrug transporter. Biochim Biophys Acta 1191:384–388PubMedCrossRefGoogle Scholar
  11. 11.
    Homolya L, Hollo Z, Germann UA, Pastan I, Gottesman MM, Sarkadi B (1993) Fluorescent cellular indicators are extruded by the multidrug resistance protein. J Biol Chem 268:21493–21496PubMedGoogle Scholar
  12. 12.
    Homolya L, Hollo M, Muller M, Mechetner EB, Sarkadi B (1996) A new method for a quantitative assessment of P-glycoprotein-related multidrug resistance in tumour cells. Br J Cancer 73:849–855PubMedGoogle Scholar
  13. 13.
    Hay RJ (1988) The seed stock concept and quality control for cell lines, Anal Biochem 171:225–237PubMedCrossRefGoogle Scholar
  14. 14.
    Hubensack M (2005) Approaches to overcome the blood–brain barrier in the chemotherapy of primary and secondary brain tumors: modulation of P-glycoprotein 170 and targeting of the transferrin receptor. Doctoral thesis, University of Regensburg,
  15. 15.
    Kemper EM, van Zandbergen AE, Cleypool C, Mos HA, Boogerd W, Beijnen JH, van Tellingen O (2003) Increased penetration of paclitaxel into the brain by inhibition of P-glycoprotein. Clin Cancer Res 9:2849–2855PubMedGoogle Scholar
  16. 16.
    Fellner S, Bauer B, Miller DS, Schaffrik M, Fankhänel M, Spruß T, Bernhardt G, Graeff C, Färber L, Gschaidmeier H, Buschauer A, Fricker G (2002) Transport of paclitaxel (Taxol) across the blood–brain barrier in vitro and in vivo. J Clin Invest 110:1309–1318PubMedCrossRefGoogle Scholar
  17. 17.
    Peters T, Lindenmaier H, Haefeli WE, Weiss J (2006) Interaction of the mitotic kinesin Eg5 inhibitor monastrol with P-glycoprotein. Naunyn Schmiedebergs Arch Pharmacol 372:291–299PubMedCrossRefGoogle Scholar
  18. 18.
    Tarby CM, Kaltenbach RF, III, Huynh T, Pudzianowski A, Shen H, Ortega-Nanos M, Sheriff S, Newitt JA, McDonnel PA, Burford N, Fairchild CR, Vaccaro W, Chen Z, Borzilleri RM, Naglich J, Lombardo LJ, Gottardis M, Trainor GL, Roussel DL (2006) Inhibitors of human mitotic kinesin Eg5: characterization of the 4-phenyl-tetrahydroisoquinoline lead series. Bioorg Med Chem Lett 16:2095–2100PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Christine Müller
    • 1
  • Dietmar Gross
    • 1
  • Vasiliki Sarli
    • 2
  • Michael Gartner
    • 2
  • Athanassios Giannis
    • 2
  • Günther Bernhardt
    • 1
  • Armin Buschauer
    • 1
  1. 1.Institute of PharmacyUniversity of RegensburgRegensburgGermany
  2. 2.Institute of Organic ChemistryUniversity of LeipzigLeipzigGermany

Personalised recommendations