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Polyvalent dendrimer glucosamine conjugates prevent scar tissue formation

Nature Biotechnology volume 22, pages 977–984 (2004)Cite this article

Abstract

Dendrimers are hyperbranched macromolecules that can be chemically synthesized to have precise structural characteristics. We used anionic, polyamidoamine, generation 3.5 dendrimers to make novel water-soluble conjugates of D(+)-glucosamine and D(+)-glucosamine 6-sulfate with immuno-modulatory and antiangiogenic properties respectively. Dendrimer glucosamine inhibited Toll-like receptor 4–mediated lipopolysaccharide induced synthesis of pro-inflammatory chemokines (MIP-1α, MIP-1β, IL-8) and cytokines (TNF-α, IL-1β, IL-6) from human dendritic cells and macrophages but allowed upregulation of the costimulatory molecules CD25, CD80, CD83 and CD86. Dendrimer glucosamine 6-sulfate blocked fibroblast growth factor-2 mediated endothelial cell proliferation and neoangiogenesis in human Matrigel and placental angiogenesis assays. When dendrimer glucosamine and dendrimer glucosamine 6-sulfate were used together in a validated and clinically relevant rabbit model of scar tissue formation after glaucoma filtration surgery, they increased the long-term success of the surgery from 30% to 80% (P = 0.029). We conclude that synthetically engineered macromolecules such as the dendrimers described here can be tailored to have defined immuno-modulatory and antiangiogenic properties, and they can be used synergistically to prevent scar tissue formation.

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Figure 1: Polyvalent dendrimer glucosamine conjugates.
Figure 2: Inhibitory effect of dendrimer glucosamine on LPS-mediated chemokine and cytokine release (a–f) Dendrimer glucosamine-inhibited pro-inflammatory chemokine (MIP-1α, MIP-1β, IL-8) (a–c) and cytokine (TNF-α, IL-1β, IL-6) (d–f) release from human PBMCs when added 30 min before, or 2 h or 4 h after LPS.
Figure 3: Cell type specific effect of dendrimer glucosamine.
Figure 4: Effect of dendrimer glucosamine on dendritic cell–mediated functions.
Figure 5: The anti-angiogenic activity of dendrimer glucosamine 6-sulfate.
Figure 6: Histological cross-sections of rabbit eyes at day 30.

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Acknowledgements

This study was supported by grants to S.S. from the National Institutes of Health (1-R21-A144694-01), The Wellcome Trust (068309) and The Wolfson Foundation (PR/013217). P.K. is supported, in part, by Moorfields Trustees, The Michael & Ilse Katz Foundation and an Alcon Research Institute Award.

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Authors and Affiliations

  1. Faculty of Medicine, Imperial College London, Hammersmith Hospital, Ducane Road, London, W12 0NN, UK

    Sunil Shaunak, Sharyn Thomas, Elisabetta Gianasi & Ian Teo

  2. Biomedical Polymers Group, School of Pharmacy, 29-39 Brunswick Square, London, WC1N 1AX, UK

    Antony Godwin & Steve Brocchini

  3. Wound Healing Research Unit, Institute of Ophthalmology, 11-43 Bath Street, London, EC1V 9EL, UK

    Emma Jones, Kamiar Mireskandari & Peng Khaw

  4. Department of Histopathology, Institute of Ophthalmology, 11-43 Bath Street, London, EC1V 9EL, UK

    Philip Luthert

  5. Center for Polymer Therapeutics, Welsh School of Pharmacy, Cardiff University, Redwood Building, King Edward VII Avenue, Cardiff, CF10 3XF, UK

    Ruth Duncan

  6. Faculty of Medicine, Imperial College London, Chelsea & Westminster Hospital, 369 Fulham Road, London, SW10 9NH, UK

    Steve Patterson

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Correspondence to Sunil Shaunak.

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Competing interests

S.S., E.G. and R.D. are inventors on a patent in which the US National Institutes of Health have an interest. This patent has been assigned to a new university spinout company called Polytherics in which The Wellcome Trust, Imperial College London, University College London, School of Pharmacy London, S.B. and S.S. hold equity.

Supplementary information

Supplementary Notes

Detailed conjugation and characterisation procedures (PDF 105 kb)

Supplementary Table 1

Matrigel assay (PDF 84 kb)

Supplementary Fig. 1

Glaucoma filtration surgery (PDF 140 kb)

Supplementary Fig. 2

Multigene chemokine plasmid for RT-PCR (PDF 297 kb)

Supplementary Methods

Real-time mRNA PCR for human chemokines and cytokines (PDF 100 kb)

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Shaunak, S., Thomas, S., Gianasi, E. et al. Polyvalent dendrimer glucosamine conjugates prevent scar tissue formation. Nat Biotechnol 22, 977–984 (2004). https://doi.org/10.1038/nbt995

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