A non-cytotoxic dendrimer with innate and potent anticancer and anti-metastatic activities

  • Nature Biomedical Engineering 1745757 (2017)
  • doi:10.1038/s41551-017-0130-9
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The structural perfection and multivalency of dendrimers have made them useful for biodelivery and bioactivity via peripheral functionalization and the modulation of core-forming structures and dendrimer generations. Yet only few dendrimers have shown inherent therapeutic activity arising from their inner repeating units. Here, we report the synthesis and characterization of a polyacylthiourea dendrimer with inherent potent anticancer activity and the absence of cytotoxicity in mice. The poly(ethylene glycol)-functionalized fourth generation of the dendrimer, which can be efficiently synthesized from sequential click reactions of orthogonal monomers, displays low in vivo acute and subacute toxicities yet potently inhibits tumour growth and metastasis. The dendrimer’s in vivo anticancer activity arises from the depletion of bioavailable copper and the subsequent inhibition of angiogenesis and cellular proliferation. When compared with some clinically used cytotoxin drugs, the dendrimer exerts inherent anticancer activity via non-cytotoxic pathways and leads to higher therapeutic efficacy, yet without cytotoxin-induced side effects.

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  1. 1.

    Kolate, A. et al. PEG—a versatile conjugating ligand for drugs and drug delivery systems. J. Control. Release 192, 67–81 (2014).

  2. 2.

    Kopeček, J. Polymer–drug conjugates: origins, progress to date and future directions. Adv. Drug Del. Rev. 65, 49–59 (2013).

  3. 3.

    Zelikin, A. N., Ehrhardt, C. & Healy, A. M. Materials and methods for delivery of biological drugs. Nat. Chem. 8, 997–1007 (2016).

  4. 4.

    Cabral, H. & Kataoka, K. Progress of drug-loaded polymeric micelles into clinical studies. J. Control. Release 190, 465–476 (2014).

  5. 5.

    Shi, J. J., Kantoff, P. W., Wooster, R. & Farokhzad, O. C. Cancer nanomedicine: progress, challenges and opportunities. Nat. Rev. Cancer 17, 20–37 (2017).

  6. 6.

    Werle, M. Natural and synthetic polymers as inhibitors of drug efflux pumps. Pharm. Res. 25, 500–511 (2008).

  7. 7.

    Sosnik, A. Reversal of multidrug resistance by the inhibition of ATP-binding cassette pumps employing “Generally Recognized As Safe” (GRAS) nanopharmaceuticals: a review. Adv. Drug Del. Rev. 65, 1828–1851 (2013).

  8. 8.

    Alakhova, D. Y. & Kabanov, A. V. Pluronics and MDR reversal: an update. Mol. Pharm. 11, 2566–2578 (2014).

  9. 9.

    Thota, B. N. S., Urner, L. H. & Haag, R. Supramolecular architectures of dendritic amphiphiles in water. Chem. Rev. 116, 2079–2102 (2016).

  10. 10.

    Hsu, H.-J., Bugno, J., Lee, S.-R. & Hong, S. Dendrimer-based nanocarriers: a versatile platform for drug delivery. Wiley. Interdiscip. Rev. Nanomed. Nanobiotechnol. 9, e1409 (2017).

  11. 11.

    Khandare, J., Calderon, M., Dagia, N. M. & Haag, R. Multifunctional dendritic polymers in nanomedicine: opportunities and challenges. Chem. Soc. Rev. 41, 2824–2848 (2012).

  12. 12.

    Wei, T. et al. Anticancer drug nanomicelles formed by self-assembling amphiphilic dendrimer to combat cancer drug resistance. Proc. Natl Acad. Sci. USA 112, 2978–2983 (2015).

  13. 13.

    Chauhan, A. S., Diwan, P. V., Jain, N. K. & Tomalia, D. A. Unexpected in vivo anti-inflammatory activity observed for simple, surface functionalized poly(amidoamine) dendrimers. Biomacromolecules 10, 1195–1202 (2009).

  14. 14.

    Dernedde, J. et al. Dendritic polyglycerol sulfates as multivalent inhibitors of inflammation. Proc. Natl Acad. Sci. USA 107, 19679–19684 (2010).

  15. 15.

    Rele, S. M. et al. Dendrimer-like PEO glycopolymers exhibit anti-inflammatory properties. J. Am. Chem. Soc. 127, 10132–10133 (2005).

  16. 16.

    Portevin, D. et al. Regulatory activity of azabisphosphonate-capped dendrimers on human CD4+ T cell proliferation enhances ex-vivo expansion of NK cells from PBMCs for immunotherapy. J. Transl. Med. 7, 82 (2009).

  17. 17.

    Hayder, M. et al. A phosphorus-based dendrimer targets inflammation and osteoclastogenesis in experimental arthritis. Sci. Transl. Med. 3, 81ra35 (2011).

  18. 18.

    Price, C. F. et al. SPL7013 Gel (VivaGel®) retains potent HIV-1 and HSV-2 inhibitory activity following vaginal administration in humans. PLoS ONE 6, e24095 (2011).

  19. 19.

    Dufès, C. et al. Synthetic anticancer gene medicine exploits intrinsic antitumor activity of cationic vector to cure established tumors. Cancer Res. 65, 8079–8084 (2005).

  20. 20.

    Al-Jamal, K. T. et al. Systemic antiangiogenic activity of cationic poly-l-lysine dendrimer delays tumor growth. Proc. Natl Acad. Sci. USA 107, 3966–3971 (2010).

  21. 21.

    Ciepluch, K. et al. Biological properties of new viologen-phosphorus dendrimers. Mol. Pharm. 9, 448–457 (2012).

  22. 22.

    Abdel-Rahman, M. A. & Al-Abd, A. M. Thermoresponsive dendrimers based on oligoethylene glycols: design, synthesis and cytotoxic activity against MCF-7 breast cancer cells. Eur. J. Med. Chem. 69, 848–854 (2013).

  23. 23.

    Sliwkowski, M. X. & Mellman, I. Antibody therapeutics in cancer. Science 341, 1192–1198 (2013).

  24. 24.

    Scott, A. M., Wolchok, J. D. & Old, L. J. Antibody therapy of cancer. Nat. Rev. Cancer 12, 278–287 (2012).

  25. 25.

    Duncan, R. Polymer therapeutics: top 10 selling pharmaceuticals—What next? J. Control. Release 190, 371–380 (2014).

  26. 26.

    Maeda, H., Nakamura, H. & Fang, J. The EPR effect for macromolecular drug delivery to solid tumors: improvement of tumor uptake, lowering of systemic toxicity, and distinct tumor imaging in vivo. Adv. Drug Del. Rev. 65, 71–79 (2013).

  27. 27.

    Dubacheva, G. V. et al. Superselective targeting using multivalent polymers. J. Am. Chem. Soc. 136, 1722–1725 (2014).

  28. 28.

    Tito, N. B. & Frenkel, D. Optimizing the selectivity of surface-adsorbing multivalent polymers. Macromolecules 47, 7496–7509 (2014).

  29. 29.

    Zhang, W. et al. Redox-hypersensitive organic nanoparticles for selective treatment of cancer cells. Chem. Mater. 28, 4440–4446 (2016).

  30. 30.

    Barner-Kowollik, C. et al. “Clicking” polymers or just efficient linking: What is the difference? Angew. Chem. Int. Ed. 50, 60–62 (2011).

  31. 31.

    Xiao, S., Turkyilmaz, S. & Smith, B. D. Convenient synthesis of multivalent zinc(II)–dipicolylamine complexes for molecular recognition. Tetrahedron Lett. 54, 861–864 (2013).

  32. 32.

    Nair, D. P. et al. The thiol-Michael addition click reaction: a powerful and widely used tool in materials chemistry. Chem. Mater. 26, 724–744 (2014).

  33. 33.

    Restani, R. B. et al. Biocompatible polyurea dendrimers with pH-dependent fluorescence. Angew. Chem. Int. Ed. 51, 5162–5165 (2012).

  34. 34.

    Wang, D., Imae, T. & Miki, M. Fluorescence emission from PAMAM and PPI dendrimers. J. Colloid Interface Sci. 306, 222–227 (2007).

  35. 35.

    Wang, H., Zhao, E., Lam, J. W. Y. & Tang, B. Z. AIE luminogens: emission brightened by aggregation. Mater. Today 18, 365–377 (2015).

  36. 36.

    Zhang Yuan, W. & Zhang, Y. Nonconventional macromolecular luminogens with aggregation-induced emission characteristics. J. Polym. Sci. A Polym. Chem. 55, 560–574 (2017).

  37. 37.

    Hu, R., Leung, N. L. & Tang, B. Z. AIE macromolecules: syntheses, structures and functionalities. Chem. Soc. Rev. 43, 4494–4562 (2014).

  38. 38.

    Shen, Y. et al. Multifunctioning pH-responsive nanoparticles from hierarchical self-assembly of polymer brush for cancer drug delivery. AIChE J. 54, 2979–2989 (2008).

  39. 39.

    Krzewska, S., Pajdowski, L. & Podsiadły, H. Studies on the reaction of copper (II) with thiourea-II: the modification of bjerrum’s method. The determination of equilibrium in simultaneous redox and complexation reactions. J. Inorg. Nucl. Chem. 42, 87–88 (1980).

  40. 40.

    Campos, C., Guzmán, R., López-Fernández, E. & Casado, Á. Evaluation of the copper(II) reduction assay using bathocuproinedisulfonic acid disodium salt for the total antioxidant capacity assessment: the CUPRAC–BCS assay. Anal. Biochem. 392, 37–44 (2009).

  41. 41.

    El Brahmi, N. et al. Original multivalent copper(II)-conjugated phosphorus dendrimers and corresponding mononuclear copper(II) complexes with antitumoral activities. Mol. Pharm. 10, 1459–1464 (2013).

  42. 42.

    Jain, S. et al. Tetrathiomolybdate-associated copper depletion decreases circulating endothelial progenitor cells in women with breast cancer at high risk of relapse. Ann. Oncol. 24, 1491–1498 (2013).

  43. 43.

    Sun, Q. et al. Integration of nanoassembly functions for an effective delivery cascade for cancer drugs. Adv. Mater. 26, 7615–7621 (2014).

  44. 44.

    Chen, L. et al. Rejection of metastatic 4T1 breast cancer by attenuation of Treg cells in combination with immune stimulation. Mol. Ther. 15, 2194–2202 (2007).

  45. 45.

    Larive, R. M. et al. Contribution of the R-Ras2 GTP-binding protein to primary breast tumorigenesis and late-stage metastatic disease. Nat. Commun. 5, 3881 (2014).

  46. 46.

    Hua, K.-T. et al. N-α-Acetyltransferase 10 protein suppresses cancer cell metastasis by binding PIX proteins and inhibiting Cdc42/Rac1 activity. Cancer Cell 19, 218–231 (2011).

  47. 47.

    Gao, D. et al. Endothelial progenitor cells control the angiogenic switch in mouse lung metastasis. Science 319, 195–198 (2008).

  48. 48.

    Longmire, M., Choyke, P. L. & Kobayashi, H. Clearance properties of nano-sized particles and molecules as imaging agents: considerations and caveats. Nanomedicine 3, 703–717 (2008).

  49. 49.

    Olivares, M., Méndez, M. A., Astudillo, P. A. & Pizarro, F. Present situation of biomarkers for copper status. Am. J. Clin. Nutr. 88, 859S–862S (2008).

  50. 50.

    Kalliolias, G. D. & Ivashkiv, L. B. TNF biology, pathogenic mechanisms and emerging therapeutic strategies. Nat. Rev. Rheumatol. 12, 49–62 (2016).

  51. 51.

    Solinas, A. et al. Acylthiourea, acylurea, and acylguanidine derivatives with potent Hedgehog inhibiting activity. J. Med. Chem. 55, 1559–1571 (2012).

  52. 52.

    Antoni, P. et al. Pushing the limits for thiol-ene and CuAAC reactions: synthesis of a 6th generation dendrimer in a single day. Macromolecules 43, 6625–6631 (2010).

  53. 53.

    Lee, C. C., MacKay, J. A., Frechet, J. M. J. & Szoka, F. C. Designing dendrimers for biological applications. Nat. Biotechnol. 23, 1517–1526 (2005).

  54. 54.

    Kesharwani, P., Jain, K. & Jain, N. K. Dendrimer as nanocarrier for drug delivery. Prog. Polym. Sci. 39, 268–307 (2014).

  55. 55.

    Liu, X. X. et al. Adaptive amphiphilic dendrimer-based nanoassemblies as robust and versatile siRNA delivery systems. Angew. Chem. Int. Ed. 53, 11822–11827 (2014).

  56. 56.

    Ornelas, C. Brief timelapse on dendrimer chemistry: advances, limitations, and expectations. Macromol. Chem. Phys. 217, 149–174 (2016).

  57. 57.

    Shaunak, S. et al. Polyvalent dendrimer glucosamine conjugates prevent scar tissue formation. Nat. Biotechnol. 22, 977–984 (2004).

  58. 58.

    Lipshultz, S. E., Cochran, T. R., Franco, V. I. & Miller, T. L. Treatment-related cardiotoxicity in survivors of childhood cancer. Nat. Rev. Clin. Oncol. 10, 697–710 (2013).

  59. 59.

    Sahni, V., Choudhury, D. & Ahmed, Z. Chemotherapy-associated renal dysfunction. Nat. Rev. Nephrol. 5, 450–462 (2009).

  60. 60.

    Schiff, D., Wen, P. Y. & van den Bent, M. J. Neurological adverse effects caused by cytotoxic and targeted therapies. Nat. Rev. Clin. Oncol. 6, 596–603 (2009).

  61. 61.

    Gupte, A. & Mumper, R. J. Elevated copper and oxidative stress in cancer cells as a target for cancer treatment. Cancer Treat. Rev. 35, 32–46 (2009).

  62. 62.

    Brady, D. C. et al. Copper is required for oncogenic BRAF signalling and tumorigenesis. Nature 509, 492–496 (2014).

  63. 63.

    MacDonald, G. et al. Memo is a copper-dependent redox protein with an essential role in migration and metastasis. Sci. Signal. 7, ra56 (2014).

  64. 64.

    Martin, F. et al. Copper-dependent activation of hypoxia-inducible factor (HIF)-1: implications for ceruloplasmin regulation. Blood 105, 4613–4619 (2005).

  65. 65.

    Lazarchick, J. Update on anemia and neutropenia in copper deficiency. Curr. Opin. Hematol. 19, 58–60 (2012).

  66. 66.

    Madsen, E. & Gitlin, J. D. Copper deficiency. Curr. Opin. Gastroenterol. 23, 187–192 (2007).

  67. 67.

    Klevay, L. M. Cardiovascular disease from copper deficiency—a history. J. Nutr. 130, 489S–492S (2000).

  68. 68.

    Olivares, M. & Uauy, R. Copper as an essential nutrient. Am. J. Clin. Nutr. 63, 791S–796S (1996).

  69. 69.

    Volm, M., Koomägi, R. & Mattern, J. Prognostic value of vascular endothelial growth factor and its receptor Flt-1 in squamous cell lung cancer. Int. J. Cancer 74, 64–68 (1997).

  70. 70.

    Weidner, N., Semple, J. P., Welch, W. R. & Folkman, J. Tumor angiogenesis and metastasis—correlation in invasive breast carcinoma. N. Engl. J. Med. 324, 1–8 (1991).

  71. 71.

    Hassouneh, B. et al. Tetrathiomolybdate promotes tumor necrosis and prevents distant metastases by suppressing angiogenesis in head and neck cancer. Mol. Cancer Ther. 6, 1039–1045 (2007).

  72. 72.

    Schosinsky, K. H., Lehmann, H. P. & Beeler, M. F. Measurement of ceruloplasmin from its oxidase activity in serum by use of o-dianisidine dihydrochloride. Clin. Chem. 20, 1556–1563 (1974).

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We thank the National Basic Research Program of China (2014CB931900), the National Natural Science Foundation of China (51390481, U1501243, 51522304, 21090352 and 50888001) and the Doctoral Fund of Ministry of Education of China (20110101130007) for financial support.

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Author notes

  1. Shiqun Shao and Quan Zhou contributed equally to this work.


  1. Center for Bionanoengineering and Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, 310027, Hangzhou, China

    • Shiqun Shao
    • , Quan Zhou
    • , Jingxing Si
    • , Jianbin Tang
    • , Xiangrui Liu
    •  & Youqing Shen
  2. The Second Affiliated Hospital of School of Medicine, Zhejiang University, 310009, Hangzhou, China

    • Jingxing Si
    • , Kai Wang
    •  & Rongzhen Xu
  3. College of Pharmaceutical Science, Zhejiang University, 310058, Hangzhou, China

    • Meng Wang
    •  & Jianqing Gao


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Y.S. designed and supervised the project and wrote the manuscript with S.S.; S.S. Q.Z. and J.S. carried out all the experiments; M.W. checked the anticancer activity; X.L., J.G., R.X. and K.W. instructed the bioassays; J.T. instructed the synthesis.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Youqing Shen.

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