title: Production and characterization of a PEGylated derivative of recombinant human deoxyribonuclease I for cystic fibrosis therapy.
abstract: Recombinant human deoxyribonuclease I (rhDNase) is the mucolytic agent most widely used for the treatment of respiratory disease in cystic fibrosis. However, rhDNase is rapidly cleared from the lungs which implies a high dosing frequency and limited patient adherence. The aim of this study was to produce a long-acting PEGylated derivative of rhDNase presenting a preserved enzymatic activity. Site-specific PEGylation on the N-terminal (N-ter) leucine residue of rhDNase was achieved by reductive alkylation at acidic pH using linear 20kDa, linear 30kDa or two-arm 40kDa polyethylene glycol (PEG) propionaldehydes. Yields of mono-PEGylated products ranged between 45% and 61%. Conjugation to PEG fully preserved the secondary structure and the in vitro enzymatic activity of the native protein. These properties offer interesting perspectives for in vivo inhalation studies of the PEGylated enzyme.
authors.affiliation: Advanced Drug Delivery & Biomaterials, Louvain Drug Research Institute (LDRI), Université catholique de Louvain, Brussels 1200, Belgium.
authors.forename: Marie-Julie
authors.initials: MJ
authors.lastname: Guichard
authors.affiliation: Advanced Drug Delivery & Biomaterials, Louvain Drug Research Institute (LDRI), Université catholique de Louvain, Brussels 1200, Belgium.
authors.forename: Harshad P
authors.initials: HP
authors.lastname: Patil
authors.affiliation: Advanced Drug Delivery & Biomaterials, Louvain Drug Research Institute (LDRI), Université catholique de Louvain, Brussels 1200, Belgium.
authors.forename: Salome Juliette
authors.initials: SJ
authors.lastname: Koussoroplis
authors.affiliation: Department of Proteomic and Microbiology, Research Institute for Biosciences, University of Mons, Mons 7000, Belgium.
authors.forename: Ruddy
authors.initials: R
authors.lastname: Wattiez
authors.affiliation: Louvain Center for Toxicology and Applied Pharmacology, Institute of Experimental and Clinical Research (IREC), Université catholique de Louvain, Brussels 1200, Belgium.
authors.forename: Teresinha
authors.initials: T
authors.lastname: Leal
authors.affiliation: Advanced Drug Delivery & Biomaterials, Louvain Drug Research Institute (LDRI), Université catholique de Louvain, Brussels 1200, Belgium. Electronic address: rita.vanbever@uclouvain.be.
authors.forename: Rita
authors.initials: R
authors.lastname: Vanbever
mesh_terms: D002626:Chemistry, Pharmaceutical; D003550:Cystic Fibrosis / Q000188:drug therapy*; D003850:Deoxyribonuclease I / Q000737:chemistry* / Q000627:therapeutic use; D004339:Drug Compounding; D005100:Expectorants / Q000737:chemistry* / Q000627:therapeutic use; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry / Q000627:therapeutic use
chemical_list: D005100:Expectorants; D011994:Recombinant Proteins; D003850:Deoxyribonuclease I; C568813:dornase alfa
keywords: Cystic fibrosis; PEGylation; Polyethylene glycol (PubChem CID: 174); Recombinant human deoxyribonuclease I