Skip to main content

Advertisement

SpringerLink
Log in

A novel class of tubulin inhibitors that exhibit potent antiproliferation and in vitro vessel-disrupting activity

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

Abstract

Purpose

Since anticancer agents that interfere with microtubule function are in widespread use and have a broad spectrum of activity against both hematological malignancies and solid tumors, there is an urgent need to develop novel tubulin inhibitors with broader activities and avoiding drug resistance.

Methods and results

In this study, we describe the characterization of select lead compounds from a novel class of indazole-based tubulin inhibitors. Three lead compounds, TH-337, TH-482 and TH-494, exhibit potent antiproliferative activity against cell lines derived from human pancreatic carcinoma, human breast adenocarcinoma and human colorectal adenocarcinoma cells. The three compounds were also tested for cytotoxicity against a panel of clinically relevant drug resistant cancer cell lines that either overexpress the drug resistance pumps MDR-1, MRP-1 and BCRP-1 or have altered Topoisomerase II activity. TH-482 and -494 retained cytotoxic activities against all of the resistant cell lines tested; however, TH-337 exhibited decreased cytotoxicity in the cell line overexpressing BCRP-1, indicating that TH-337 is a substrate of that pump. We show that TH-482’s antiproliferative activity is due to cell cycle arrest at the G2/M phase. We demonstrate that TH-482 binds specifically to the colchicine site of tubulin and that it inhibits tubulin polymerization in vitro in a concentration-dependent manner. The in vitro anti-vascular activities of TH-482 were assessed using the HUVEC-C cell line. TH-482 inhibits in vitro neovessel formation and disrupts pre-established vessels using HUVEC-C cells. TH-482 also increases permeability of vascular endothelial cells in a concentration- and time-dependent manner.

Conclusions

TH-482 demonstrates potent in vitro efficacy as a novel tubulin-targeted anti-proliferative and anti-vascular agent and notably is more potent in antiproliferative assays than the benchmark compound combretastatin A-4. These results identify TH-482 as a potent tubulin inhibitor, and support the investigation of its in vivo efficacy and pharmacokinetic properties as the prototype of a new class of anti-tubulin agents.

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

Abbreviations

MDR-1:

Multi-drug resistant-1

MRP-1:

Multidrug resistance-associated protein-1

BCRP:

Breast cancer resistant protein

CA-4:

Combretastatin A-4

ITC:

Isothermal titration calorimetry

PEM:

Pipes, EGTA and MgCl2 (general tubulin buffer)

References

  1. Hadfield JA, Ducki S, Hirst N et al (2003) Tubulin and microtubules as targets for anticancer drugs. Prog Cell Cycle Res 5:309–325

    PubMed  Google Scholar 

  2. Jordan MA, Wilson L (2004) Microtubules as a target for anticancer drugs. Nat Rev Cancer 4:253–265

    Article  PubMed  CAS  Google Scholar 

  3. Yoshida D, Hoshino S, Shimura T et al (2000) Drug-induced apoptosis by anti-microtubule agent, estramustine phosphate on human malignant glioma cell line, U87MG; in vitro study. J Neurooncol 47:133–140

    Article  PubMed  CAS  Google Scholar 

  4. Mayer TU, Kapoor TM, Haggarty SJ et al (1999) Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen. Science 286:971–974

    Article  PubMed  CAS  Google Scholar 

  5. Tozer GM, Kanthou C, Baguley BC (2005) Disrupting tumour blood vessels. Nat Rev Cancer 5:423–435

    Article  PubMed  CAS  Google Scholar 

  6. Jain RK (2003) Molecular regulation of vessel maturation. Nat Med 9:685–693

    Article  PubMed  CAS  Google Scholar 

  7. Chang YS, di Tomaso E, McDonald DM et al (2000) Mosaic blood vessels in tumors: frequency of cancer cells in contact with flowing blood. Proc Natl Acad Sci USA 97:14608–14613

    Article  PubMed  CAS  Google Scholar 

  8. Lippert JW (2007) Vascular disrupting agents. Bioorg Med Chem 15:605–615

    Article  PubMed  CAS  Google Scholar 

  9. Nicholson B, Lloyd GK, Miller BR et al (2006) NPI-2358 is a tubulin-depolymerizing agent: in-vitro evidence for activity as a tumor vascular-disrupting agent. Anticancer Drugs 17:25–31

    Article  PubMed  CAS  Google Scholar 

  10. Peifer C, Stoiber T, Unger E et al (2006) Design, synthesis, and biological evaluation of 3,4-diarylmaleimides as angiogenesis inhibitors. J Med Chem 49:1271–1281

    Article  PubMed  CAS  Google Scholar 

  11. Davis PD, Dougherty GJ, Blakey DC et al (2002) ZD6126: a novel vascular-targeting agent that causes selective destruction of tumor vasculature. Cancer Res 62:7247–7253

    PubMed  CAS  Google Scholar 

  12. Gottesman MM, Fojo T, Bates SE (2002) Multidrug resistance in cancer: role of ATP-dependent transporters. Nat Rev Cancer 2:48–58

    Article  PubMed  CAS  Google Scholar 

  13. Szakacs G, Annereau JP, Lababidi S et al (2004) Predicting drug sensitivity and resistance: profiling ABC transporter genes in cancer cells. Cancer Cell 6:129–137

    Article  PubMed  CAS  Google Scholar 

  14. Szakacs G, Paterson JK, Ludwig JA et al (2006) Targeting multidrug resistance in cancer. Nat Rev Drug Discov 5:219–234

    Article  PubMed  CAS  Google Scholar 

  15. Lee JS, Paull K, Alvarez M et al (1994) Rhodamine efflux patterns predict P-glycoprotein substrates in the National Cancer Institute drug screen. Mol Pharmacol 46:627–638

    PubMed  CAS  Google Scholar 

  16. Shoemaker RH (2000) Genetic and epigenetic factors in anticancer drug resistance. J Natl Cancer Inst 92:4–5

    Article  PubMed  CAS  Google Scholar 

  17. Goncalves A, Braguer D, Kamath K et al (2001) Resistance to Taxol in lung cancer cells associated with increased microtubule dynamics. Proc Natl Acad Sci USA 98:11737–11742

    Article  PubMed  CAS  Google Scholar 

  18. Giannakakou P, Sackett DL, Kang YK et al (1997) Paclitaxel-resistant human ovarian cancer cells have mutant beta-tubulins that exhibit impaired paclitaxel-driven polymerization. J Biol Chem 272:17118–17125

    Article  PubMed  CAS  Google Scholar 

  19. Zhang CC, Yang JM, White E et al (1998) The role of MAP4 expression in the sensitivity to paclitaxel and resistance to vinca alkaloids in p53 mutant cells. Oncogene 16:1617–1624

    Article  PubMed  CAS  Google Scholar 

  20. Kavallaris M, Kuo DY, Burkhart CA et al (1997) Taxol-resistant epithelial ovarian tumors are associated with altered expression of specific beta-tubulin isotypes. J Clin Invest 100:1282–1293

    Article  PubMed  CAS  Google Scholar 

  21. Duan JX, Cai X, Meng F et al (2007) Potent antitubulin tumor cell cytotoxins based on 3-aroyl indazoles. J Med Chem 50:1001–1006

    Article  PubMed  CAS  Google Scholar 

  22. Robey RW, Honjo Y, Morisaki K et al (2003) Mutations at amino-acid 482 in the ABCG2 gene affect substrate and antagonist specificity. Br J Cancer 89:1971–1978

    Article  PubMed  CAS  Google Scholar 

  23. Meng F, Nguyen XT, Cai X et al (2007) ARC-111 inhibits hypoxia-mediated hypoxia-inducible factor-1 alpha accumulation. Anticancer Drugs 18:435–445

    Article  PubMed  CAS  Google Scholar 

  24. Bonne D, Heusele C, Simon C et al (1985) 4′,6-Diamidino-2-phenylindole, a fluorescent probe for tubulin and microtubules. J Biol Chem 260:2819–2825

    PubMed  CAS  Google Scholar 

  25. Cushman M, Nagarathnam D, Gopal D et al (1992) Synthesis and evaluation of analogues of (Z)-1-(4-methoxyphenyl)-2-(3,4,5-trimethoxyphenyl)ethene as potential cytotoxic and antimitotic agents. J Med Chem 35:2293–2306

    Article  PubMed  CAS  Google Scholar 

  26. Harker WG, Sikic BI (1985) Multidrug (pleiotropic) resistance in doxorubicin-selected variants of the human sarcoma cell line MES-SA. Cancer Res 45:4091–4096

    PubMed  CAS  Google Scholar 

  27. Hipfner DR, Gauldie SD, Deeley RG et al (1994) Detection of the M(r) 190,000 multidrug resistance protein, MRP, with monoclonal antibodies. Cancer Res 54:5788–5792

    PubMed  CAS  Google Scholar 

  28. Harker WG, Slade DL, Drake FH et al (1991) Mitoxantrone resistance in HL-60 leukemia cells: reduced nuclear topoisomerase II catalytic activity and drug-induced DNA cleavage in association with reduced expression of the topoisomerase II beta isoform. Biochemistry 30:9953–9961

    Article  PubMed  CAS  Google Scholar 

  29. Ruben AJ, Kiso Y, Freire E (2006) Overcoming roadblocks in lead optimization: a thermodynamic perspective. Chem Biol Drug Des 67:2–4

    Article  PubMed  CAS  Google Scholar 

  30. Franklin TJ, Jacobs V, Jones G et al (1996) Glucuronidation associated with intrinsic resistance to mycophenolic acid in human colorectal carcinoma cells. Cancer Res 56:984–987

    PubMed  CAS  Google Scholar 

  31. Lichtner RB, Rotgeri A, Bunte T et al (2001) Subcellular distribution of epothilones in human tumor cells. Proc Natl Acad Sci USA 98:11743–11748

    Article  PubMed  CAS  Google Scholar 

  32. Jordan MA, Wendell K, Gardiner S et al (1996) Mitotic block induced in HeLa cells by low concentrations of paclitaxel (Taxol) results in abnormal mitotic exit and apoptotic cell death. Cancer Res 56:816–825

    PubMed  CAS  Google Scholar 

  33. Gwaltney SL 2nd, Imade HM, Barr KJ et al (2001) Novel sulfonate analogues of combretastatin A-4: potent antimitotic agents. Bioorg Med Chem Lett 11:871–874

    Article  PubMed  CAS  Google Scholar 

  34. Romagnoli R, Baraldi PG, Remusat V et al (2006) Synthesis and biological evaluation of 2-(3′,4′,5′-trimethoxybenzoyl)-3-amino 5-aryl thiophenes as a new class of tubulin inhibitors. J Med Chem 49:6425–6428

    Article  PubMed  CAS  Google Scholar 

  35. Romagnoli R, Baraldi PG, Pavani MG et al (2006) Synthesis and biological evaluation of 2-amino-3-(3′,4′,5′-trimethoxybenzoyl)-5-aryl thiophenes as a new class of potent antitubulin agents. J Med Chem 49:3906–3915

    Article  PubMed  CAS  Google Scholar 

  36. Shan B, Medina JC, Santha E et al (1999) Selective, covalent modification of beta-tubulin residue Cys-239 by T138067, an antitumor agent with in vivo efficacy against multidrug-resistant tumors. Proc Natl Acad Sci USA 96:5686–5691

    Article  PubMed  CAS  Google Scholar 

  37. Tron GC, Pirali T, Sorba G et al (2006) Medicinal chemistry of combretastatin A4: present and future directions. J Med Chem 49:3033–3044

    Article  PubMed  CAS  Google Scholar 

  38. Helmlinger G, Netti PA, Lichtenbeld HC et al (1997) Solid stress inhibits the growth of multicellular tumor spheroids. Nat Biotechnol 15:778–783

    Article  PubMed  CAS  Google Scholar 

  39. Tozer GM, Prise VE, Wilson J et al (1999) Combretastatin A-4 phosphate as a tumor vascular-targeting agent: early effects in tumors and normal tissues. Cancer Res 59: 1626–1634

    PubMed  CAS  Google Scholar 

  40. Baguley BC, Holdaway KM, Thomsen LL et al (1991) Inhibition of growth of colon 38 adenocarcinoma by vinblastine and colchicine: evidence for a vascular mechanism. Eur J Cancer 27:482–487

    Article  PubMed  CAS  Google Scholar 

  41. Hill SA, Lonergan SJ, Denekamp J et al (1993) Vinca alkaloids: anti-vascular effects in a murine tumour. Eur J Cancer 29A:1320–1324

    Article  PubMed  CAS  Google Scholar 

  42. Tozer GM, Prise VE, Wilson J et al (2001) Mechanisms associated with tumor vascular shut-down induced by combretastatin A-4 phosphate: intravital microscopy and measurement of vascular permeability. Cancer Res 61:6413–6422

    PubMed  CAS  Google Scholar 

  43. Gifford SM, Grummer MA, Pierre SA et al (2004) Functional characterization of HUVEC-CS: Ca2+ signaling, ERK 1/2 activation, mitogenesis and vasodilator production. J Endocrinol 182:485–499

    Article  PubMed  CAS  Google Scholar 

  44. Vincent L, Kermani P, Young LM et al (2005) Combretastatin A4 phosphate induces rapid regression of tumor neovessels and growth through interference with vascular endothelial-cadherin signaling. J Clin Invest 115:2992–3006

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Threshold Pharmaceuticals, Inc., 1300 Seaport Blvd, Redwood City, CA, 94063, USA

    Fanying Meng, Xiaohong Cai, Jianxin Duan, Mark G. Matteucci & Charles P. Hart

Authors
  1. Fanying Meng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaohong Cai
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jianxin Duan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mark G. Matteucci
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Charles P. Hart
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Fanying Meng.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Meng, F., Cai, X., Duan, J. et al. A novel class of tubulin inhibitors that exhibit potent antiproliferation and in vitro vessel-disrupting activity. Cancer Chemother Pharmacol 61, 953–963 (2008). https://doi.org/10.1007/s00280-007-0549-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00280-007-0549-x

Keywords

Search

Navigation