Targeted PEG-poly(glutamic acid) complexes for inhalation protein delivery to the lung

Nieto-Orellana, A.; Li, H.; Rosiere, R.; Wauthoz, N.; Williams, H.; Monteiro, C.J.; Bosquillon, C.; Childerhouse, N.; Keegan, G.; Coghlan, D.; Mantovani, G.; Stolnik, S.

Abstract

Pulmonary delivery is increasingly seen as an attractive, non-invasive route for the delivery of forthcoming protein therapeutics. In this context, here we describe protein complexes with a new ‘complexing excipient’ - vitamin B12-targeted poly(ethylene glycol)-block-poly(glutamic acid) copolymers. These form complexes in sub-200 nm size with a model protein, suitable for cellular targeting and intracellular delivery. Initially we confirmed expression of vitamin B12-internalization receptor (CD320) by Calu-3 cells of the in vitro lung epithelial model used, and demonstrated enhanced B12 receptor-mediated cellular internalization of B12-targeted complexes, relative to non-targeted counterparts or protein alone. To develop an inhalation formulation, the protein complexes were spray dried adopting a standard protocol into powders with aerodynamic diameter within the suitable range for lower airway deposition. The cellular internalization of targeted complexes from dry powders applied directly to Calu-3 model was found to be 2–3 fold higher compared to non-targeted complexes. The copolymer complexes show no complement activation, and in vivo lung tolerance studies demonstrated that repeated administration of formulated dry powders over a 3 week period in healthy BALB/c mice induced no significant toxicity or indications of lung inflammation, as assessed by cell population count and quantification of IL-1β, IL-6, and TNF-α pro-inflammatory markers. Importantly, the in vivo data appear to suggest that B12-targeted polymer complexes administered as dry powder enhance lung retention of their protein payload, relative to protein alone and non-targeted counterparts. Taken together, our data illustrate the potential developability of novel B12-targeted poly(ethylene glycol)-poly(glutamic acid) copolymers as excipients suitable to be formulated into a dry powder product for the inhalation delivery of proteins, with no significant lung toxicity, and with enhanced protein retention at their in vivo target tissue.

Citation

Nieto-Orellana, A., Li, H., Rosiere, R., Wauthoz, N., Williams, H., Monteiro, C., …Stolnik, S. (2019). Targeted PEG-poly(glutamic acid) complexes for inhalation protein delivery to the lung. Journal of Controlled Release, 316, 250-262. https://doi.org/10.1016/j.jconrel.2019.10.012

• Journal Article Type: Article
• Acceptance Date: Oct 6, 2019
• Online Publication Date: Oct 31, 2019
• Publication Date: Dec 28, 2019
• Deposit Date: Nov 15, 2019
• Publicly Available Date: Nov 1, 2020
• Journal: Journal of Controlled Release
• Print ISSN: 0168-3659
• Electronic ISSN: 1873-4995
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 316
• Pages: 250-262
• DOI: https://doi.org/10.1016/j.jconrel.2019.10.012
• Keywords: Polymer-protein complexes; Inhalation delivery; Spray-dried inhalation powder; Protein delivery; Targeted complexes
• Public URL: https://nottingham-repository.worktribe.com/output/3032652
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0168365919305747
• Contract Date: Nov 15, 2019