Cationic host defence peptides constitute a promising class of therapeutic drug leads with a wide range of therapeutic applications, including anticancer therapy, immunomodulation, and antimicrobial activity. Although potent and efficacious, systemic toxicity and low chemical stability have hampered their commercial development. To overcome these challenges a novel nanogel-based drug delivery system was designed.
The peptide novicidin was self-assembled with an octenyl succinic anhydride-modified analogue of hyaluronic acid, and this formulation was optimized using a microfluidics-based quality-by-design approach.
By applying design-of-experiment it was demonstrated that the encapsulation efficiency of novicidin (15% to 71%) and the zeta potential (−24 to −57 mV) of the nanogels could be tailored by changing the preparation process parameters, with a maximum peptide loading of 36 ± 4%. The nanogels exhibited good colloidal stability under different ionic strength conditions and allowed complete release of the peptide over 14 days. Furthermore, self-assembly of novicidin with hyaluronic acid into nanogels significantly improved the safety profile at least five-fold and six-fold when tested in HUVECs and NIH 3T3 cells, respectively, whilst showing no loss of antimicrobial activity against Escherichia coli and Staphylococcus aureus.
Formulation in nanogels could be a viable approach to improve the safety profile of host defence peptides.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Price excludes VAT (USA)
Tax calculation will be finalised during checkout.
Dynamic light scattering
Dulbecco’s modified Eagle’s medium
Design of experiment
Fetal bovine serum
Hank’s balanced salt solution
Host defense peptides
Human umbilical vein endothelial cell
Minimum inhibitory concentration
Multiple linear regression
Nanoparticle tracking analysis
Octenyl succinic anhydride-modified hyaluronic acid
Transmission electron microscopy
Zasloff M. Antimicrobial peptides of multicellular organisms. Nature. 2002;415:389–95.
Nguyen LT, Haney EF, Vogel HJ. The expanding scope of antimicrobial peptide structures and their modes of action. Trends Biotechnol. 2011;29:464–72.
Huang W, Seo J, Willingham SB, Czyzewski AM, Gonzalgo ML, Weissman IL, et al. Learning from host-defense peptides: cationic, amphipathic peptoids with potent anticancer activity. PLoS ONE. 2014;9:e90397.
Gaspar D, Veiga AS, Castanho MARB. From antimicrobial to anticancer peptides. A review. Front Microbiol. 2013;4:294.
Mulder KCL, Lima LA, Miranda VJ, Dias SC, Franco OL. Current scenario of peptide-based drugs: the key roles of cationic antitumor and antiviral peptides. Front Microbiol. 2013;4:321.
Hilchie AL, Wuerth K, Hancock REW. Immune modulation by multifaceted cationic host defense (antimicrobial) peptides. Nat Chem Biol. 2013;9:761–8.
Mayer ML, Blohmke CJ, Falsafi R, Fjell CD, Madera L, Turvey SE, et al. Rescue of dysfunctional autophagy attenuates hyperinflammatory responses from cystic fibrosis cells. J Immunol. 2013;190:1227–38.
Hancock RE. Cationic peptides: effectors in innate immunity and novel antimicrobials. Lancet Infect Dis. 2001;1:156–64.
Giuliani A, Pirri G, Nicoletto SF. Antimicrobial peptides: an overview of a promising class of therapeutics. Cent Eur J Biol. 2007;2:1–33.
Hancock REW, Sahl H-G. Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies. Nat Biotechnol. 2006;24:1551–7.
Van’t Hof W, Veerman EC, Helmerhorst EJ, Amerongen AV. Antimicrobial peptides: properties and applicability. Biol Chem. 2001;382:597–619.
Nochi T, Yuki Y, Takahashi H, Sawada S, Mejima M, Kohda T, et al. Nanogel antigenic protein-delivery system for adjuvant-free intranasal vaccines. Nat Mater. 2010;9:572–8.
Yan M, Ge J, Liu Z, Ouyang P. Encapsulation of single enzyme in nanogel with enhanced biocatalytic activity and stability. J Am Chem Soc. 2006;128:11008–9.
Takahashi H, Sawada S, Akiyoshi K. Amphiphilic polysaccharide nanoballs: a new building block for nanogel biomedical engineering and artificial chaperones. ACS Nano. 2010;5:337–45.
Moghimi SM, Hunter AC, Murray JC. Long-circulating and target-specific nanoparticles: theory to practice. Pharmacol Rev. 2001;53:283–318.
Peppas NA, Bures P, Leobandung W, Ichikawa H. Hydrogels in pharmaceutical formulations. Eur J Pharm Biopharm. 2000;50:27–46.
Capretto L, Cheng W, Hill M, Zhang X. Micromixing within microfluidic devices. Top Curr Chem. 2011;304:27–68.
Jahn A, Reiner JE, Vreeland WN, DeVoe DL, Locascio LE, Gaitan M. Preparation of nanoparticles by continuous-flow microfluidics. J Nanoparticle Res. 2008;10:925–34.
De Smedt SC, Demeester J, Hennink WE. Cationic polymer based gene delivery systems. Pharm Res. 2000;17:113–26.
Jeong JH, Park TG, Kim SH. Self-assembled and nanostructured siRNA delivery systems. Pharm Res. 2011;28:2072–85.
Balakrishnan VS, Vad BS, Otzen DE. Novicidin’s membrane permeabilizing activity is driven by membrane partitioning but not by helicity: a biophysical study of the impact of lipid charge and cholesterol. Biochim Biophys Acta. 2013;1834:996–1002.
Dorosz J, Gofman Y, Kolusheva S, Otzen D, Ben-Tal N, Nielsen NC, et al. Membrane interactions of novicidin, a novel antimicrobial peptide: phosphatidylglycerol promotes bilayer insertion. J Phys Chem B. 2010;114:11053–60.
Gottlieb CT, Thomsen LE, Ingmer H, Mygind PH, Kristensen H-H, Gram L. Antimicrobial peptides effectively kill a broad spectrum of Listeria monocytogenes and Staphylococcus aureus strains independently of origin, sub-type, or virulence factor expression. BMC Microbiol. 2008;8:205.
Hoskin DW, Ramamoorthy A. Studies on anticancer activities of antimicrobial peptides. Biochim Biophys Acta. 2008;1778:357–75.
Vandamme D, Landuyt B, Luyten W, Schoofs L. A comprehensive summary of LL-37, the factotum human cathelicidin peptide. Cell Immunol. 2012;280:22–35.
Eenschooten C, Guillaumie F, Kontogeorgis GM, Stenby EH, Schwach-Abdellaoui K. Preparation and structural characterisation of novel and versatile amphiphilic octenyl succinic anhydride–modified hyaluronic acid derivatives. Carbohydr Polym. 2010;79:597–605.
Ossipov DA. Nanostructured hyaluronic acid-based materials for active delivery to cancer. Expert Opin Drug Deliv. 2010;7:681–703.
Kim Y, Lee Chung B, Ma M, Mulder WJM, Fayad ZA, Farokhzad OC, et al. Mass production and size control of lipid-polymer hybrid nanoparticles through controlled microvortices. Nano Lett. 2012;12:3587–91.
Cory AH, Owen TC, Barltrop JA, Cory JG. Use of an aqueous soluble tetrazolium/formazan assay for cell growth assays in culture. Cancer Commun. 1991;3:207–12.
CLSI. Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically ; approved standard—ninth edition. CLSI document M07-A9. Wayne: Clinical and Laboratory Standards Institute; 2012.
Kim Y, Fay F, Cormode DP, Sanchez-Gaytan BL, Tang J, Hennessy EJ, et al. Single step reconstitution of multifunctional high-density lipoprotein-derived nanomaterials using microfluidics. ACS Nano. 2013;7:9975–83.
Karayianni M, Pispas S, Chryssikos GD, Gionis V, Giatrellis S, Nounesis G. Complexation of lysozyme with poly(sodium(sulfamate-carboxylate)isoprene). Biomacromolecules. 2011;12:1697–706.
Slaninová J, Mlsová V, Kroupová H, Alán L, Tůmová T, Monincová L, et al. Toxicity study of antimicrobial peptides from wild bee venom and their analogs toward mammalian normal and cancer cells. Peptides. 2012;33:18–26.
Park JH, Cho HJ, Yoon HY, Yoon IS, Ko SH, Shim JS, et al. Hyaluronic acid derivative-coated nanohybrid liposomes for cancer imaging and drug delivery. J Control Release. 2014;174:98–108.
He M, Zhao Z, Yin L, Tang C, Yin C. Hyaluronic acid coated poly(butyl cyanoacrylate) nanoparticles as anticancer drug carriers. Int J Pharm. 2009;373:165–73.
Zhang W, Cheng Q, Guo S, Lin D, Huang P, Liu J, et al. Gene transfection efficacy and biocompatibility of polycation/DNA complexes coated with enzyme degradable PEGylated hyaluronic acid. Biomaterials. 2013;34:6495–503.
Yang XY, Li YX, Li M, Zhang L, Feng LX, Zhang N. Hyaluronic acid-coated nanostructured lipid carriers for targeting paclitaxel to cancer. Cancer Lett. 2013;334:338–45.
Tossi A, Sandri L, Giangaspero A. Amphipathic, α-helical antimicrobial peptides. Biopolymers. 2000;55:4–30.
ACKNOWLEDGMENTS AND DISCLOSURES
The authors acknowledge Prof. Robert Langer at MIT for his generous support and discussion on the use of microfluidic devices. We also acknowledge Karina Juul Vissing, Thara Qais Hussein and Maria Læssøe Pedersen for their technical support. Pall Thor Ingvarsson, PhD, is acknowledged for assistance with the experimental design; The Danish Agency for Science and Technology and Innovation (DanCARD, grant no. 06-097075) for financial support, The Core Facility for Integrated Microscopy, Faculty of Health and Medical Sciences, University of Copenhagen for providing access to imaging facilities and Adam Bohr, PhD, for his assistance. Lastly, Innovation Fund Denmark (041-2010-3) is acknowledged for co-financing the HPLC system.
About this article
Cite this article
Water, J.J., Kim, Y., Maltesen, M.J. et al. Hyaluronic Acid-Based Nanogels Produced by Microfluidics-Facilitated Self-Assembly Improves the Safety Profile of the Cationic Host Defense Peptide Novicidin. Pharm Res 32, 2727–2735 (2015). https://doi.org/10.1007/s11095-015-1658-6