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Inhibition of MDR1 activity and induction of apoptosis by analogues of nifedipine and diltiazem: an in vitro analysis

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Summary

We report herein the reversal of multidrug resistance-1 (MDR1) in A2780/DX3 cells by the two nifedipine-like compounds 1 and 2 that are part of a library of 1,4-dihydropyridines (1,4-DHPs) calcium-channel modulators bearing in C-4 a different substituted imidazo[2,1-b]thiazole system. By methylthiazol tetrazolium (MTT) assay, cytofluorimetry, and fluorescence microscopy we evaluated their ability to reverse MDR in our cell system. Moreover, together with compound 3 (the diltiazem-like 8-(4-chlorophenyl)-5-methyl-8-[(2Z)-pent-2-en-1-yloxy]-8H-[1,2,4]oxadiazolo[3,4-c][1,4]thiazin-3-one) we analyzed their ability to potentiate the triggering of apoptosis after exposure to doxorubicin, through the nuclear morphological analysis after 4′,6-diamidino-2-phenylindole (DAPI), the fluorescein isothiocyanate (FITC)-Annexin-V/propidium iodide (PI) staining and the caspase activity determination. Our results demonstrate that compounds 1 and 2, at concentrations showing a very low (5%) or absent inhibition of cell proliferation, in combination with doxorubicin enhance its antiproliferative activity (from 30% to 54% IC50 reduction) in A2780/DX3 cells through an increase of doxorubicin intracellular accumulation. These compounds together with compound 3, which has already been demonstrated to act as a potent inhibitor of MDR1 function, were also able to significantly potentiate the activation of the apoptosis machinery triggered by the exposure to doxorubicin. In conclusion, our results identify two new molecules structurally related to the calcium-channel blocker nifedipine, but characterized by a very low LTCC blockers activity, able to potentiate the antiproliferative and apoptotic activities of doxorubicin through an increase of its intracellular concentration likely caused by the inhibition of MDR1 function.

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Acknowledgements

This research was supported by funds from “Ministero dell’Istruzione, dell’Università e della Ricerca” (FIRB 2001, PRIN-2005034305_001, PRIN-20078J9L2A_005) and from the University of Bologna. Dr. M. Croce is recipient of a fellowship by “Fondazione italiana per la lotta al neuroblastoma”.

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Authors and Affiliations

  1. Istituto Nazionale per la Ricerca sul Cancro, S.C. Terapia Immunologica, L.go R. Benzi 10, 16132, Genova, Italy

    Maurizio Viale, Daniela de Totero, Cinzia Aiello & Michela Croce

  2. Dipartimento di Chimica Organica “A. Mangini”, Università degli Studi di Bologna, Via San Giacomo 11, 40126, Bologna, Italy

    Cinzia Cordazzo

  3. Dipartimento di Scienze Farmaceutiche, Università degli Studi di Bologna, Via Belmeloro 6, 40126, Bologna, Italy

    Roberta Budriesi, Alberto Leoni, Pierfranco Ioan, Aldo Andreani, Mirella Rambaldi & Alberto Chiarini

  4. Istituto Nazionale per la Ricerca sul Cancro, S.C. Nanobiotecnologie, L.go R. Benzi 10, 16132, Genova, Italy

    Camillo Rosano

  5. Istituto Nazionale per la Ricerca sul Cancro, S.S. Tumori Polmonari, L.go R. Benzi 10, 16132, Genova, Italy

    Patrizia Russo

  6. Dipartimento di Chimica “G. Ciamician”, Università degli Studi di Bologna, Via Selmi 2, 40126, Bologna, Italy

    Domenico Spinelli

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  1. Maurizio Viale
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Correspondence to Maurizio Viale or Domenico Spinelli.

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Viale, M., Cordazzo, C., de Totero, D. et al. Inhibition of MDR1 activity and induction of apoptosis by analogues of nifedipine and diltiazem: an in vitro analysis. Invest New Drugs 29, 98–109 (2011). https://doi.org/10.1007/s10637-009-9340-7

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  • DOI: https://doi.org/10.1007/s10637-009-9340-7

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