Skip to main content

Advertisement

SpringerLink
Log in

Novel microtubule-targeting agents, pyrrolo-1,5-benzoxazepines, induce apoptosis in multi-drug-resistant cancer cells

  • Original Article
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

Purpose

The development of multi-drug resistance (MDR) due to the expression of members of the ATP binding cassette (ABC) transporter family is a major obstacle in cancer treatment. The broad range of substrate specificities associated with these transporters leads to the efflux of many anti-cancer drugs from tumour cells. Therefore, the development of new chemotherapeutic agents that are not substrates of these transporters is important. We have recently demonstrated that some members of a novel series of pyrrolo-1,5-benzoxazepine (PBOX) compounds are microtubule-depolymerising agents that potently induce apoptosis in several cancer cell lines and impair growth of mouse breast tumours. The aim of this current study was to establish whether PBOXs were capable of inducing apoptosis in cancer cells expressing either P-glycoprotein or breast cancer resistance protein (BCRP), two of the main ABC transporters associated with MDR.

Methods

We performed in vitro studies to assess the effects of PBOXs on cell proliferation, cell cycle and apoptosis in human cancer cell lines and their drug-resistant substrains expressing either P-glycoprotein or BCRP. In addition, we performed a preliminary molecular docking study to examine interactions between PBOXs and P-glycoprotein.

Results

We established that three representative PBOXs, PBOX-6, -15 and -16 were capable of inducing apoptosis in drug-resistant HL60-MDR1 cells (expressing P-glycoprotein) and HL60-ABCG2 cells (expressing BCRP) with similar potencies as in parental human promyelocytic leukaemia HL60 cells. Likewise, resistance to PBOX-6 and -16 was not evident in P-glycoprotein-expressing A2780-ADR cells in comparison with parent human ovarian carcinoma A2780 cells. Finally, we deduced by molecular docking that PBOX-6 is not likely to form favourable interactions with the substrate binding site of P-glycoprotein.

Conclusion

Our results suggest that pro-apoptotic PBOX compounds may be potential candidates for the treatment of P-glycoprotein- or BCRP-associated MDR cancers.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Mulligan J, Greene LM, Cloonan S et al (2006) Identification of tubulin as the molecular target of pro-apoptotic pyrrolo-1, 5-benzoxazepines. Mol Pharmacol 70:60–70

    CAS  PubMed  Google Scholar 

  2. Zisterer DM, Campiani G, Nacci V et al (2000) Pyrrolo-1, 5-benzoxazepines induce apoptosis in HL-60, Jurkat, and Hut-78 cells: a new class of apoptotic agents. J Pharmacol Exp Ther 293:48–59

    CAS  PubMed  Google Scholar 

  3. Mc Gee MM, Campiani G, Ramunno A et al (2001) Pyrrolo-1, 5-benzoxazepines induce apoptosis in chronic myelogenous leukemia (CML) cells by bypassing the apoptotic suppressor bcr-abl. J Pharmacol Exp Ther 296:31–40

    CAS  PubMed  Google Scholar 

  4. Mc Gee MM, Hyland E, Campiani G et al (2002) Caspase-3 is not essential for DNA fragmentation in MCF-7 cells during apoptosis induced by the pyrrolo-1, 5-benzoxazepine, PBOX-6. FEBS Lett 515:66–70

    Article  CAS  PubMed  Google Scholar 

  5. Mc Gee MM, Campiani G, Ramunno A et al (2002) Activation of the c-Jun N-terminal kinase (JNK) signaling pathway is essential during PBOX-6-induced apoptosis in chronic myelogenous leukemia (CML) cells. J Biol Chem 277:18383–18389

    Article  CAS  PubMed  Google Scholar 

  6. Mc Gee MM, Greene LM, Ledwidge S et al (2004) Selective induction of apoptosis by the pyrrolo-1, 5-benzoxazepine 7-[[dimethylcarbamoyl]oxy]-6-(2-naphthyl)pyrrolo-[2, 1-d] (1, 5)-benzoxazepine (PBOX-6) in Leukemia cells occurs via the c-Jun NH2-terminal kinase-dependent phosphorylation and inactivation of Bcl-2 and Bcl-XL. J Pharmacol Exp Ther 310:1084–1095

    Article  CAS  PubMed  Google Scholar 

  7. McElligott AM, Maginn EN, Greene LM et al (2009) The novel tubulin targeting agent, pyrrolo-1, 5-benzoxazepine-15, induces apoptosis in poor prognostic subgroups of Chronic Lymphocytic Leukemia. Cancer Res 69:8366–8375

    Article  CAS  PubMed  Google Scholar 

  8. Nathwani SM, Butler S, Meegan MJ et al (2010) Dual targeting of tumour cells and host endothelial cells by novel microtubule-targeting agents, pyrrolo-1, 5-benzoxazepines. Cancer Chemother Pharmacol 65:289–300

    Article  CAS  Google Scholar 

  9. Greene LM, Fleeton M, Mulligan J et al (2005) The pyrrolo-1, 5-benzoxazepine, PBOX-6, inhibits the growth of breast cancer cells in vitro independent of estrogen receptor status, and inhibits breast tumour growth in vivo. Oncol Rep 5:1357–1363

    Google Scholar 

  10. Mashima T, Seimiya H, Chen Z et al (1998) Apoptosis resistance in tumour cells. Cytotechnology 27:293–308

    Article  CAS  PubMed  Google Scholar 

  11. Cowan KH, Batist G, Tulpule A et al (1986) Similar biochemical changes associated with multidrug resistance in human breast cancer cells and carcinogen-induced resistance to xenobiotics in rats. Proc Natl Acad Sci USA 83:9328–9332

    Article  CAS  PubMed  Google Scholar 

  12. Redmond SM, Joncourt F, Buser K et al (1991) Assessment of P-glycoprotein, gluthathione-based detoxifying enzymes and O6-alkylguanine-DNA alkyltransferase as potential indicators of constitutive drug resistance in human colorectal tumours. Cancer Res 51:2092–2097

    CAS  PubMed  Google Scholar 

  13. Eijdems EW, de Haas M, Timmerman AJ et al (1995) Reduced topoisomerase II activity in multidrug-resistant human non-small cell lung cancer cell lines. Br J Cancer 71:40–47

    CAS  PubMed  Google Scholar 

  14. Sarkadi B, Homolya L, Szakacs G et al (2006) Human multidrug resistance ABCB and ABCG transporters: participation in a chemoimmunity defence system. Physiol Rev 86:1179–1236

    Article  CAS  PubMed  Google Scholar 

  15. Maliepaard M, Scheffer GL, Faneyte IF et al (2001) Subcellular localisation and distribution of the breast cancer resistance protein transporter in normal human tissues. Cancer Res 61:3458–3464

    CAS  PubMed  Google Scholar 

  16. Scheffer GL, Maliepaard M, Pijnenborg AC (2000) Breast cancer resistance protein is localized at the plasma membrane in mitoxantrone- and topotecan-resistant cell lines. Cancer Res 60:2589–2593

    CAS  PubMed  Google Scholar 

  17. Juliano RL, Ling V (1976) A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. Biochim Biophys Acta 455:152–162

    Article  CAS  PubMed  Google Scholar 

  18. Zaman GJR, Flens MJ, van Leusden MR et al (1994) The human multidrug resistance-associated protein MRP is a plasma membrane drug-efflux pump. Proc Nat Acad Sci 91:8822–8826

    Article  CAS  PubMed  Google Scholar 

  19. Doyle LA, Yang W, Abruzzo LV et al (1998) A multidrug resistance transporter from human MCF-7 breast cancer cells. Proc Natl Acad Sci USA 95:15665–15670

    Article  CAS  PubMed  Google Scholar 

  20. Sarkadi B, Ozvegy-Laczka C, Nemet K et al (2004) ABCG2—a transporter for all seasons. FEBS Lett 567:116–120

    Article  CAS  PubMed  Google Scholar 

  21. Nakatomi K, Yoshikawa M, Oka M (2001) Transport of 7-ethyl-10-hydroxycamptothecin (SN-38) by breast cancer resistance protein ABCG2 in human lung cancer cells. Biochem Biophys Res Commun 288:827–832

    Article  CAS  PubMed  Google Scholar 

  22. Schellens JH, Maliepaard M, Scheper RJ et al (2000) Transport of topoisomerase I inhibitors by the breast cancer resistance protein. Potential clinical implications. Ann NY Acad Sci 922:188–194

    Article  CAS  PubMed  Google Scholar 

  23. Volk EL, Schneider E (2003) Wild-type breast cancer resistance protein (BCRP/ABCG2) is a methotrexate polyglutamate transporter. Cancer Res 63:5538–5543

    CAS  PubMed  Google Scholar 

  24. Borowski E, Bontemps-Gracz MM, Piwkowska A (2005) Strategies for overcoming ABC-transporters-mediated multidrug resistance (MDR) of tumour cells. Acta Biochim Pol 52:609–627

    CAS  PubMed  Google Scholar 

  25. Ozvegy-Laczka C, Hegedus T, Varady G et al (2004) High-affinity interaction of tyrosine kinase inhibitors with ABCG2 multidrug transporter. Mol Pharmacol 65:1485–1495

    Article  PubMed  Google Scholar 

  26. Campiani G, Nacci V, Fiorini I et al (1996) Synthesis, biological activity, and SARs of pyrrolobenzoxazepine derivatives, a new class of specific “peripheral-type” benzodiazepine receptor ligands. J Med Chem 39:3435–3450

    Article  CAS  PubMed  Google Scholar 

  27. Penzotti JE, Lamb ML, Evensen E et al (2002) A computational ensemble pharmacophore model for identifying substrates of P-glycoprotein. J Med Chem 45:1737–1740

    Article  CAS  PubMed  Google Scholar 

  28. Aller SG, Yu J, Ward A et al (2009) Structure of P-glycoprotein reveals a molecular basis for poly-specific drug binding. Science 323:1718–1722

    Article  CAS  PubMed  Google Scholar 

  29. Greene LM, Campiani G, Lawler M et al (2008) BubR1 is required for a sustained mitotic spindle checkpoint arrest in human cancer cells treated with tubulin-targeting pyrrolo-1, 5-benzoxazepines. Mol Pharmacol 73:419–430

    Article  CAS  PubMed  Google Scholar 

  30. Verma NK, Dempsey E, Conroy J et al (2008) A new microtubule-targeting compound PBOX-15 inhibits T-cell migration via post-translational modifications of tubulin. J Mol Med 86:457–469

    Article  CAS  PubMed  Google Scholar 

  31. Greene LM, Kelly L, Onnis V et al (2007) STI-571 enhances the apoptotic efficacy of PBOX-6, a novel microtubule targeting agent, in both STI-571-sensitive and -resistant Bcr-Abl-positive human chronic myeloid leukemia cells. J Pharmacol Exp Ther 321:288–297

    Article  CAS  PubMed  Google Scholar 

  32. Mollinedo F, Gajate C (2003) Microtubules, microtubule-interfering agents and apoptosis. Apoptosis 8:413–450

    Article  CAS  PubMed  Google Scholar 

  33. Toffoli G, Simone F, Corona G et al (1995) Structure activity relationship of verapamil analogues and reversal of multidrug resistance. Biochem Pharmacol 50:1245–1255

    Article  CAS  PubMed  Google Scholar 

  34. Rabindran SK, He H, Singh M et al (1998) Reversal of a novel multidrug resistance mechanism in human colon carcinoma cells by fumitremorgan C. Cancer Res 58:5850–5858

    CAS  PubMed  Google Scholar 

  35. Rabindran SK, Ross DD, Doyle LA et al (2000) Fumitremorgan C reverses drug resistance in cells transfected with breast cancer resistance protein. Cancer Res 60:47–50

    CAS  PubMed  Google Scholar 

  36. Seelig A, Landwojtowicz E (2000) Structure-activity relationship of P-glycoprotein substrates and modifiers. Eur J Pharm Sci 12:31–40

    Article  CAS  PubMed  Google Scholar 

  37. Morris PG, Fornier MN (2008) Microtubule active agents: beyond the frontier. Clin Cancer Res 14:7167–7172

    Article  CAS  PubMed  Google Scholar 

  38. Bacher G, Nickel B, Emig P et al (2001) D-24851, a novel synthetic microtubule inhibitor, exerts curative antitumoral activity in vivo, shows efficacy toward multidrug-resistant tumour cells, and lacks neurotoxicity. Cancer Res 61:392–399

    CAS  PubMed  Google Scholar 

  39. Wienecke A, Bacher G (2009) Indibulin, a novel microtubule inhibitor, discriminates between mature neuronal and nonneuronal tubulin. Cancer Res 69:171–177

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This project was kindly funded by Science Foundation Ireland. In addition, the Molecular Design Group gratefully acknowledges the generous academic support afforded by the Chemical Computing Group (MOE software), OpenEye Scientific (OpenEye product suite), The Trinity Centre for High Performance Computing and Accelrys (Pipeline Pilot) in the delivery of this work.

Author information

Authors and Affiliations

  1. School of Biochemistry and Immunology, Trinity College, Dublin 2, Ireland

    Seema-Maria Nathwani, Naomi N. McGovern, D. Clive Williams & Daniela M. Zisterer

  2. School of Pharmacy and Pharmaceutical Sciences, Trinity College, Dublin 2, Ireland

    Stephen Butler & Mary J. Meegan

  3. Molecular Design Group, School of Biochemistry and Immunology, Trinity College, Dublin 2, Ireland

    Darren Fayne & David G. Lloyd

  4. Membrane Research Group of Hungarian Academy of Sciences, Semmelweis University and National Blood Center, Budapest, Hungary

    Balazs Sarkadi

  5. European Research Centre for Drug Discovery and Development, Banchi di Sotto 55 and Dipartimento Farmaco Chimico Tecnologico, Universita’ di Siena, via Aldo Moro 2, 53100, Siena, Italy

    Giuseppe Campiani

  6. Institute of Molecular Medicine, St. James’s Hospital and Trinity College, Dublin, Ireland

    Mark Lawler

Authors
  1. Seema-Maria Nathwani
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stephen Butler
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Darren Fayne
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Naomi N. McGovern
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Balazs Sarkadi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mary J. Meegan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. David G. Lloyd
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Giuseppe Campiani
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Mark Lawler
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. D. Clive Williams
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Daniela M. Zisterer
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Seema-Maria Nathwani.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nathwani, SM., Butler, S., Fayne, D. et al. Novel microtubule-targeting agents, pyrrolo-1,5-benzoxazepines, induce apoptosis in multi-drug-resistant cancer cells. Cancer Chemother Pharmacol 66, 585–596 (2010). https://doi.org/10.1007/s00280-009-1200-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00280-009-1200-9

Keywords

Search

Navigation