title: Enhanced antisense efficacy of oligonucleotides adsorbed to monomethylaminoethylmethacrylate methylmethacrylate copolymer nanoparticles.
abstract: The purpose of this study was the investigation of cationic nanoparticles as drug delivery systems for antisense oligonucleotides. Cationic monomethylaminoethylmethacrylate (MMAEMA) copolymer nanoparticles were prepared from N-monomethylaminoethylmethacrylate hydrochloride and methylmethacrylate. Oligonucleotides were adsorbed onto MMAEMA nanoparticles. Cell penetration was investigated in vitro with fluorescently labeled oligonucleotides and nanoparticles. Antisense effects of oligonucleotides adsorbed to MMAEMA nanoparticles were evaluated by sequence specific inhibition of ecto-5'-nucleotidase expression. The amount of enzyme expressed in PC12 cells was detected and quantified by immunocytochemistry using fluorescein isothiocyanate-labeled antibodies. Oligonucleotides were adsorbed to MMAEMA nanoparticles by the formation of ion-pairs between the positively charged secondary amino groups located on the particle surface and the anionic phosphodiester or phosphorothioate backbones of the oligonucleotides. Adsorption to nanoparticles led to an increased cellular uptake of oligonucleotides and to a significantly enhanced antisense efficacy of unmodified phosphodiester oligonucleotides as well as phosphorothioates. The results of the cell penetration and the antisense assay demonstrated that MMAEMA nanoparticles are promising carriers for oligonucleotide administration.
authors.affiliation: Institut für Pharmazeutische Technologie, Biozentrum, Johann Wolfgang Goethe-Universität, Frankfurt am Main, Germany.
authors.forename: H P
authors.initials: HP
authors.lastname: Zobel
authors.forename: M
authors.initials: M
authors.lastname: Junghans
authors.forename: V
authors.initials: V
authors.lastname: Maienschein
authors.forename: D
authors.initials: D
authors.lastname: Werner
authors.forename: M
authors.initials: M
authors.lastname: Gilbert
authors.forename: H
authors.initials: H
authors.lastname: Zimmermann
authors.forename: C
authors.initials: C
authors.lastname: Noe
authors.forename: J
authors.initials: J
authors.lastname: Kreuter
authors.forename: A
authors.initials: A
authors.lastname: Zimmer
mesh_terms: D015720:5'-Nucleotidase / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D000327:Adsorption; D000818:Animals; D007150:Immunohistochemistry; D008689:Methacrylates; D008863:Microspheres; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000378:metabolism / Q000494:pharmacology*; D016716:PC12 Cells; D011108:Polymers; D051381:Rats
chemical_list: D008689:Methacrylates; D016376:Oligonucleotides, Antisense; D011108:Polymers; D015720:5'-Nucleotidase