Cystic fibrosis; Electrospraying; PEGylation; Recombinant human deoxyribonuclease I (rhDNase); Thermodynamic stability; Vibrating mesh nebulization; dornase alfa; Polyethylene Glycols; Humans; Animals; Mice; Feasibility Studies; Nebulizers and Vaporizers; Administration, Inhalation; Polyethylene Glycols/therapeutic use; Respiratory Aerosols and Droplets; Cystic Fibrosis/drug therapy; Pharmaceutical Science
Abstract :
[en] Recombinant human deoxyribonuclease I (rhDNase, Pulmozyme®) is the most frequently used mucolytic agent for the symptomatic treatment of cystic fibrosis (CF) lung disease. Conjugation of rhDNase to polyethylene glycol (PEG) has been shown to greatly prolong its residence time in the lungs and improve its therapeutic efficacy in mice. To present an added value over current rhDNase treatment, PEGylated rhDNase needs to be efficiently and less frequently administrated by aerosolization and possibly at higher concentrations than existing rhDNase. In this study, the effects of PEGylation on the thermodynamic stability of rhDNase was investigated using linear 20 kDa, linear 30 kDa and 2-armed 40 kDa PEGs. The suitability of PEG30-rhDNase to electrohydrodynamic atomization (electrospraying) as well as the feasibility of using two vibrating mesh nebulizers, the optimized eFlow® Technology nebulizer (eFlow) and Innospire Go, at varying protein concentrations were investigated. PEGylation was shown to destabilize rhDNase upon chemical-induced denaturation and ethanol exposure. Yet, PEG30-rhDNase was stable enough to withstand aerosolization stresses using the eFlow and Innospire Go nebulizers even at higher concentrations (5 mg of protein per ml) than conventional rhDNase formulation (1 mg/ml). High aerosol output (up to 1.5 ml per min) and excellent aerosol characteristics (up to 83% fine particle fraction) were achieved while preserving protein integrity and enzymatic activity. This work demonstrates the technical feasibility of PEG-rhDNase nebulization with advanced vibrating membrane nebulizers, encouraging further pharmaceutical and clinical developments of a long-acting PEGylated alternative to rhDNase for treating patients with CF.
Disciplines :
Pharmacy, pharmacology & toxicology
Author, co-author :
Mahri, Sohaib; Université catholique de Louvain (UCLouvain), Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials, Brussels, Belgium
Wilms, Tobias; Université catholique de Louvain (UCLouvain), Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials, Brussels, Belgium
Hagedorm, Paul; University of Groningen, Groningen Research Institute of Pharmacy, Pharmaceutical Technology and Biopharmacy, Groningen, the Netherlands
Guichard, Marie-Julie; Université catholique de Louvain (UCLouvain), Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials, Brussels, Belgium
Vanvarenberg, Kevin; Université catholique de Louvain (UCLouvain), Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials, Brussels, Belgium
Frijlink, Henderik; University of Groningen, Groningen Research Institute of Pharmacy, Pharmaceutical Technology and Biopharmacy, Groningen, the Netherlands
Vanbever, Rita ; Université catholique de Louvain (UCLouvain), Louvain Drug Research Institute, Advanced Drug Delivery and Biomaterials, Brussels, Belgium. Electronic address: rita.vanbever@uclouvain.be
Language :
English
Title :
Nebulization of PEGylated recombinant human deoxyribonuclease I using vibrating membrane nebulizers: A technical feasibility study.
This work was supported by research grants from the First Spin-off program of the Belgian Walloon Region (Grant 1910026) and Laboratoires SMB (Brussels, Belgium). We thank PARI for providing the optimized eFlow nebulizer device. Mireille Dumoulin and Rita Vanbever are respectively Research Associate and Research Director of the Fonds National de la Recherche Scientifique (F.R.S.-FNRS, Belgium).This work was supported by research grants from the First Spin-off program of the Belgian Walloon Region (Grant 1910026 ) and Laboratoires SMB (Brussels, Belgium). We thank PARI for providing the optimized eFlow nebulizer device. Mireille Dumoulin and Rita Vanbever are respectively Research Associate and Research Director of the Fonds National de la Recherche Scientifique (F.R.S.-FNRS, Belgium).
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