Pharmaceutical Research

, Volume 29, Issue 11, pp 3033–3039 | Cite as

Novel Water-Soluble Substituted Pyrrolo[3,2-d]pyrimidines: Design, Synthesis, and Biological Evaluation as Antitubulin Antitumor Agents

  • Aleem Gangjee
  • Roheeth K. Pavana
  • Wei Li
  • Ernest Hamel
  • Cara Westbrook
  • Susan L. Mooberry
Research Paper



To study the effects of a regioisomeric change on the biological activities of previously reported water soluble, colchicine site binding, microtubule depolymerizing agents.


Nine pyrrolo[3,2-d]pyrimidines were designed and synthesized. The importance of various substituents was evaluated. Their abilities to cause cellular microtubule depolymerization, inhibit proliferation of MDA-MB-435 tumor cells and inhibit colchicine binding to tubulin were studied. One of the compounds was also evaluated in the National Cancer Institute preclinical 60 cell line panel.


Pyrrolo[3,2-d]pyrimidine analogs were more potent than their pyrrolo[2,3-d]pyrimidine regioisomers. We identified compounds with submicromolar potency against cellular proliferation. The structure-activity relationship study gave insight into substituents that were crucial for activity and those that improved activity. The compound tested in the NCI 60 cell line is a 2-digit nanomolar (GI50) inhibitor of 8 tumor cell lines.


We have identified substituted pyrrolo[3,2-d]pyrimidines that are water-soluble colchicine site microtubule depolymerizing agents. These compounds serve as leads for further optimization.


antitubulin colchicine-site binders drug design microtubule depolymerizer pyrrolo[3,2-d]pyrimidines 


Acknowledgments and Disclosures

National Cancer Institute for performing the in vitro antitumor evaluation in their 60 tumor preclinical screening program.

Grant from the National Institute of Health, National Cancer Institute, CA142868 (AG,SLM).

NSF equipment grant for the NMR (NMR: CHE 0614785) and the CTRC Cancer Center Support Grant, P30 CA054174


  1. 1.
    Gangjee A, Zhao Y, Lin L, Raghavan S, Roberts EG, Risinger AL, et al. Synthesis and discovery of water-soluble microtubule targeting agents that bind to the colchicine site on tubulin and circumvent Pgp mediated resistance. J Med Chem. 2010;53(22):8116–28.PubMedCrossRefGoogle Scholar
  2. 2.
    Gangjee A, Zhao Y, Lin L, Raghavan S, Roberts EG, Risinger AL, et al. Corrections to synthesis and discovery of water-soluble microtubule targeting agents that bind to the colchicine site on tubulin and circumvent Pgp mediated resistance. J Med Chem 2011;54(3):913.Google Scholar
  3. 3.
    Leonard GD, Fojo T, Bates SE. The role of ABC transporters in clinical practice. Oncologist. 2003;8:411–24.PubMedCrossRefGoogle Scholar
  4. 4.
    Ling V. Multidrug resistance: molecular mechanisms and clinical relevance. Cancer Chemother Pharmacol. 1997;40:S3–8.PubMedCrossRefGoogle Scholar
  5. 5.
    Ferrandina G, Zannoni GF, Martinelli E, Paglia A, Gallotta V, Mozzetti S, et al. Class III β-tubulin overexpression is a marker of poor clinical outcome in advanced ovarian cancer patients. Clin Cancer Res. 2006;12:2774–9.PubMedCrossRefGoogle Scholar
  6. 6.
    Mozzetti S, Ferlini C, Concolino P, Filippetti F, Raspaglio G, Prislei S, et al. Class III β-tubulin overexpression is a prominent mechanism of paclitaxel resistance in ovarian cancer patients. Clin Cancer Res. 2005;11:298–305.PubMedGoogle Scholar
  7. 7.
    Rosell R, Scagliotti G, Danenberg KD, Lord RVN, Bepler G, Novello S, et al. Transcripts in pretreatment biopsies from a three-arm randomized trial in metastatic non-small-cell lung cancer. Oncogene. 2003;22:3548–53.PubMedCrossRefGoogle Scholar
  8. 8.
    Seve P, Isaac S, Tredan O, Souquet P-J, Pacheco Y, Perol M, et al. Expression of class III β-tubulin is predictive of patient outcome in patients with non-small cell lung cancer receiving vinorelbine-based chemotherapy. Clin Cancer Res. 2005;11:5481–6.PubMedCrossRefGoogle Scholar
  9. 9.
    Tommasi S, Mangia A, Lacalamita R, Bellizzi A, Fedele V, Chiriatti A, et al. Cytoskeleton and paclitaxel sensitivity in breast cancer: the role of β-tubulins. Int J Cancer. 2007;120:2078–85.PubMedCrossRefGoogle Scholar
  10. 10.
    Sizova OS, Modnikova GA, Glushkov RG, Solov'eva NP, Ryabokon NA, Chernov VA, et al. Synthesis and biological activity of 4,7-substituted pyrrolo[3,2-d]pyrimidines. Khim-Farm Zh. 1984;18:958–62.Google Scholar
  11. 11.
    Gangjee A, Li W, Yang J, Kisliuk RL. Design, synthesis, and biological evaluation of classical and nonclassical 2-amino-4-oxo-5-substituted-6-methylpyrrolo[3,2-d]pyrimidines as dual thymidylate synthase and dihydrofolate reductase inhibitors. J Med Chem. 2008;51(1):68–76.PubMedCrossRefGoogle Scholar
  12. 12.
    Haraguchi K, Horii C, Yoshimura Y, Ariga F, Tadokoro A, Tanaka H. An access to the β-anomer of 4'-thio-c-ribonucleosides: Hydroboration of 1-C-aryl- or 1-C-heteroaryl-4-thiofuranoid glycals and its regiochemical outcome. J Org Chem. 2011;76:8658–69.PubMedCrossRefGoogle Scholar
  13. 13.
    Stadlwieser J, Schmidt B, Bernsmann H, Dunkern T, Benediktus E, Pahl A, et al. inventors; Nycomed GmbH, Germany. assignee. Methylpyrrolopyrimidinecarboxamides as phosphodiesterase type 5 inhibitors and their preparation and use in the treatment of diseases. Patent WO2011023693A1. 2011.Google Scholar
  14. 14.
    Lee L, Robb LM, Lee M, Davis R, Mackay H, Chavda S, et al. Design, synthesis, and biological evaluations of 2,5-diaryl-2,3-dihydro-1,3,4-oxadiazoline analogs of combretastatin-A4. J Med Chem. 2010;53(1):325–34.PubMedCrossRefGoogle Scholar
  15. 15.
    Skehan P, Storeng R, Scudiero D, Monks A, McMahon J, Vistica D, et al. New colorimetric cytotoxicity assay for anticancer-drug screening. J Natl Cancer Inst. 1990;82(13):1107–12.PubMedCrossRefGoogle Scholar
  16. 16.
    Boyd MR, Paull KD. Some practical considerations and applications of the national cancer institute in vitro anticancer drug discovery screen. Drug Develop Res. 1995;34(2):91–109.CrossRefGoogle Scholar
  17. 17.
    Risinger AL, Jackson EM, Polin LA, Helms GL, LeBoeuf DA, Joe PA, et al. The taccalonolides: microtubule stabilizers that circumvent clinically relevant taxane resistance mechanisms. Cancer Res. 2008;68(21):8881–8.PubMedCrossRefGoogle Scholar
  18. 18.
    Hamel E. Evaluation of antimitotic agents by quantitative comparisons of their effects on the polymerization of purified tubulin. Cell Biochem Biophys. 2003;38(1):1–21.PubMedCrossRefGoogle Scholar
  19. 19.
    Hamel E, Lin CM. Separation of active tubulin and microtubule-associated proteins by ultracentrifugation and isolation of a component causing the formation of microtubule bundles. Biochemistry. 1984;23(18):4173–84.PubMedCrossRefGoogle Scholar
  20. 20.
    Lin CM, Ho HH, Pettit GR, Hamel E. Antimitotic natural products combretastatin A-4 and combretastatin A-2: Studies on the mechanism of their inhibition of the binding of colchicine to tubulin. Biochemistry. 1989;28(17):6984–91.PubMedCrossRefGoogle Scholar
  21. 21.
    Borisy GG. A rapid method for quantitative determination of microtubule protein using deae-cellulose filters. Anal Biochem. 1972;50(2):373–85.PubMedCrossRefGoogle Scholar
  22. 22.
    Verdier-Pinard P, Lai J-Y, Yoo H-D, Yu J, Marquez B, Nagle DG, et al. Structure-activity analysis of the interaction of curacin A, the potent colchicine site antimitotic agent, with tubulin and effects of analogs on the growth of MCF-7 breast cancer cells. Mol Pharmacol. 1998;53(1):62–76.PubMedGoogle Scholar
  23. 23.
    Hamel E, Lin CM. Stabilization of the colchicine-binding activity of tubulin by organic acids. Biochim Biophys Acta. 1981;675(2):226–31.PubMedCrossRefGoogle Scholar
  24. 24.
    Shoemaker RH. The NCI60 human tumour cell line anticancer drug screen. Nat Rev Cancer. 2006;6(10):813–23.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Aleem Gangjee
    • 1
  • Roheeth K. Pavana
    • 1
  • Wei Li
    • 1
  • Ernest Hamel
    • 2
  • Cara Westbrook
    • 3
  • Susan L. Mooberry
    • 3
    • 4
  1. 1.Division of Medicinal ChemistryGraduate School of Pharmaceutical Sciences, Duquesne UniversityPittsburghUSA
  2. 2.Screening Technologies Branch Developmental Therapeutics Program Division of Cancer Treatment & Diagnosis Frederick National Laboratory for Cancer Research National Cancer InstituteFrederickUSA
  3. 3.Department of PharmacologyUniversity of Texas Health Science Center at San AntonioSan AntonioUSA
  4. 4.Cancer Therapy & Research Center University of Texas Health Science Center at San AntonioSan AntonioUSA

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