Summary
A new semisynthetic anti-tumour bis-indol compound, KAR-2 [3′-(β-chloroethyl)-2′,4′-dioxo-3,5′-spiro-oxazolidino-4-deacetoxy-vinblastine] with lower toxicity than vinca alkaloids used in chemotherapy binds to calmodulin but, in contrast to vinblastine, does not exhibit anti-calmodulin activity. To investigate whether the modest chemical modification of bis-indol structure is responsible for the lack of anti-calmodulin potency and for the different pharmacological effects, new derivatives have been synthesized for comparative studies. The synthesis of the KAR derivatives are presented. The comparative studies showed that the spiro-oxazolidino ring and the substitution of a formyl group to a methyl one were responsible for the lack of anti-calmodulin activities. The new derivatives, similar to the mother compounds, inhibited the tubulin assembly in polymerization tests in vitro, however their inhibitory effect was highly dependent on the organization state of microtubules; bundled microtubules appeared to be resistant against the drugs. The maximal cytotoxic activities of KAR derivatives in in vivo mice hosting leukaemia P388 or Ehrlich ascites tumour cells appeared similar to that of vinblastine or vincristine, however significant prolongation of life span could be reached with KAR derivatives only after the administration of a single dose. These studies plus data obtained using a cultured human neuroblastoma cell line showed that KAR compounds displayed their cytotoxic activities at significantly higher concentrations than the mother compounds, although their antimicrotubular activities were similar in vitro. These data suggest that vinblastine/vincristine damage additional crucial cell functions, one of which could be related to calmodulin-mediated processes.
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Alley, M. A., Scudiero, D. A., Monks, A., Hursey, M. L., Czerwinski, M. J., Fine, D. L., Abbott, B. J., Mayo, J. G., Shoemaker, R. H. & Boyd, M. R. (1988). Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay. Cancer Res 48: 589–601.
Codaccioni, F., Dell’Amico, M., Boudeaux, M., Briand, C. & Lux, B. (1988). Influence of the guanine nucleotide phosphorylation state and of Mg2+ ions on the interaction of vinzolidine/tubulin 6S. Arch Biochem Biophys 267: 236–244.
Cros, S., Wright, M., Morimoto, M., Lataste, H., Couzinier, J. P. & Krikorian, A. (1989). Experimental antitumor activity of navelbine. Semin Oncol 16 (suppl 4): 15–20.
De Brabander, M., Nuydens, R., Geuens, G., Moeremans, M., De Mey, J. & Hopkins, C. (1980). Nanovid ultramicroscopy: a new non-destructive approach providing new insights in subcellular motility. In Microtubules and Microtubule Inhibitors, De Brabander M and De Mey J (eds), pp. 187–196. Elsevier: Amsterdam
De Bruyn, A., De Taeye, L., Simonds, R., Verzele, M. & De Pauw, C. (1982). Alkaloids from Catharanthus roseus. Isolation and identification of 17-desacetoxyvinblastine and 17-desacetoxyleurosine. Bull Soc Chim Belg 91: 75–85.
Durrieu, C., Bernier-Valentin, F. & Rousset, B. (1987). Binding of glyceraldehyde-3-phosphate dehydrogenase to microtubules. Mol Cell Biochem 74: 55–65.
Dustin, P. (1984). Microtubules, Springer-Verlag: Berlin
Finlay, G. J., Wilson, W. R. & Baguley, B. C. (1986). Comparison of in vitro activity of cytotoxic drugs towards human carcinoma and leukaemia cell lines. Eur J Cancer Clin Oncol 22: 655–662.
Gopalakrishna, R. & Anderson, W. B. (1982). Ca2+-induced hydrophobic site on calmodulin: application for purification of calmodulin by phenyl-sepharose affinity chromatography. Biochem Biophys Res Commun 104: 830–836.
Hesterberg, L. K. & Lee, J. C. (1982). Self-association of rabbit muscle phosphofructokinase: effects of ligands. Biochemistry 21: 216–222.
Laemmli, U. K. (1970). Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 680–688.
Lathan, B., Clark, G. M. & Von Hoff, D. D. (1985). In vitro comparison of vinzolidine and vinblastine: a model for methods of evaluation of analogues in a human tumor cloning system. Cancer Res 45: 6286–6289.
Lehotzky, A., Telegdi, M., Liliom, K. & Ovádi, J. (1993). Interaction of phosphofructokinase with tubulin and microtubule: quantitative evaluation of the mutual effects. J Biol Chem 268: 10888–10894.
Lehotzky, A., Pálfia, Z., Kovács, J., Molnár, A. & Ovádi, J. (1994). Ligand-modulated cross-bridging of microtubules by phosphofructokinase. Biochem Biophys Res Commun 204: 585–591.
Liliom, K., Lehotzky, A., Molnár, A. & Ovádi, J. (1995). Characterization of tubulin–drug interactions by ELISA. Anal Biochem 228: 18–27.
Lin, C. M., Singh, S. B., Chu, P. S., Dempcy, R. O., Schmidt, J. M., Pettit, G. R. & Hamel, E. (1988). Interactions of tubulin with potent natural and synthetic analogs of the antimitotic agent combretastin: a structure-activity study. Mol Pharmacol 34: 200–208.
Mayr, G. W. (1987). Interaction of calmodulin with phosphofructokinase: binding studies and evaluation of enzymatic and physicochemical changes. Methods Enzymol 139: 745–763.
Miller, J. C. & Antowsk, G. E. (1984). BE 861 417. Drugs Future 9: 913
Molnár, A., Liliom, K., Orosz, F., Vértessy, B. G. & Ovádi, J. (1995). Anti-calmodulin potency of indol alkaloids in in vitro systems. Eur J Pharmacol 291: 73–82.
Na, C. N. & Timasheff, S. N. (1986). Interaction of vinblastine with calf brain tubulin: multiple equilibria. Biochemistry 25: 6214–6222.
Orosz, F., Christova, T. Y. & Ovádi, J. (1988). Functional in vitro test of calmodulin antagonism: effect of drugs on interaction between calmodulin and glycolytic enzymes. Mol Pharmacol 33: 678–682.
Orosz, F., Telegdi, M., Liliom, K., Solti, M., Korbonits, D. & Ovádi, J. (1990). Dissimilar mechanisms of action of anti-calmodulin drugs: quantitative analysis. Mol Pharmacol 38: 910–916.
Orosz, F., Kovács, J., Löw, P., Vértessy, B. G., Urbányi, Z., Ács, T., Keve, T. & Ovádi, J. (1997a). Interaction of a new bis-indol derivative, KAR-2 with tubulin and its antimitotic activity. Br J Pharmacol 21: 947–954.
Orosz, F., Vértessy, B. G., Salerno, C., Crifo, C., Capuozzo, E. & Ovádi, J. (1997b). The interaction of a new anti-tumor drug, KAR-2 with calmodulin. Br J Pharmacol 21: 955–962.
Ovádi, J. (1989). Effects of drugs on calmodulin-mediated enzymatic actions. A review. Prog Drug Res 33: 353–395.
Schiff, P. B., Fant, J. & Horwitz, S. B. (1979). Promotion of microtubule assembly in vitro by taxol. Nature 277: 665–667.
Schneider, F. (1981). The aerobic glycolysis of tumor cells. Naturwissenschaften 68: 20–27.
Solti, M., Dévay, P., Kiss, I., Londesborough, J. & Friedrich, P. (1983). Cyclic nucleotide phosphodiesterases in larval and brain of wild type and dunce mutant strains of Drosophila melanogaster: isozyme pattern and activation by Ca2+/calmodulin. Biochem Biophys Res Commun 111: 652–658.
Vértessy, B. G., Kovács, J. & Ovádi, J. (1996). Specific characteristics of phosphofructokinase–microtubule interaction. FEBS Lett 379: 191–195.
Vértessy, B. G., Kovács, J., Löw, P., Lehotzky, A., Molnár, A., Orosz, F. & Ovádi, J. (1997). Characterization of microtubule-phosphofructokinase complex: specific effects of MgATP and vinblastine. Biochemistry 36: 2051–2062.
Watterson, D. M., Harrelson, W. G., Keller, P. M., Sharief, F. & Vanaman, T. C. (1976). Structural similarities between the Ca2+-dependent regulatory proteins of 3′:5′-cyclic nucleotide phosphodiesterase and actomyosin ATPase. J Biol Chem 251: 4501–4513.
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*1H and 13C chemical shift data for the sulphuric acid salts of compounds KAR-2, KAR-3 and KAR-4 will be published elsewhere.
ActivityPFK – Activity PFK–CaM
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Orosz, F., Comin, B., Raïs, B. et al. New semisynthetic vinca alkaloids: chemical, biochemical and cellular studies. Br J Cancer 79, 1356–1365 (1999). https://doi.org/10.1038/sj.bjc.6690218
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DOI: https://doi.org/10.1038/sj.bjc.6690218