Complexation to cationic microspheres of double-stranded peptide nucleic acid-DNA chimeras exhibiting decoy activity(378 views) Mischiati C, Sereni A, Finotti A, Breda L, Cortesi R, Nastruzzi C, Romanelli A, Saviano M, Bianchi N, Pedone C, Borgatti M, Gambari R
Journal Of Biomedical Science (ISSN: 1021-7770), 2004 Sep; 11(5): 697-704.
Keywords: Gene Therapy, Microspheres, Peptide Nucleic Acid-Dna Chimeras, Cell Extract, Dimethyldioctadecylammonium Bromide, Double Stranded Dna, Eudragit, Immunoglobulin Enhancer Binding Protein, Phosphorus 32, Solvent, Surfactant, Transcription Factor, Unclassified Drug, Agar Gel Electrophoresis, Article, Binding Affinity, Body Fluid, Chemical Structure, Complex Formation, Controlled Study, Electron Microscopy, Enzymatic Degradation, Evaporation, Ex Vivo Study, Human, Human Cell, Hybrid, Isotope Labeling, Molecule, Particle Size, Pharmacogenomics, Priority Journal, Simulation, Toxicity Testing, Base Sequence, Binding Sites, K562 Cells, Oligodeoxyribonucleotides,
Affiliations: *** IBB - CNR ***
Dept. of Biochem. and Molec. Biology, University of Ferrara, Ferrara, Italy
Dept. of Pharmaceutical Sciences, University of Ferrara, Ferrara, Italy
Dept. Pharmaceutical Chem. Technol., University of Perugia, Perugia, Italy
Inst. of Biostructure and Bioimaging, CNR, Napoli, Italy
Biotechnology Center, University of Ferrara, Ferrara, Italy
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Complexation to cationic microspheres of double-stranded peptide nucleic acid-DNA chimeras exhibiting decoy activity
The major aim of this paper was to determine whether cationic microspheres (CM), consisting of the permeable polymer Eudragit(R) RS 100 plus the cationic surfactant dioctadecyl-dimethyl-ammonium bromide (DDAB(18)), could bind to double-stranded peptide nucleic acid PNA-DNA-PNA (PDP) chimeras exhibiting decoy activity against NF-kappaB transcription factors. Microspheres were produced by the 'solvent evaporation method' and centrifugation at 500, 1,000 and 3,000 rpm to obtain different-sized microparticles. Microsphere morphology, size and size distribution were determined by optical and electron microscopy observations. In order to determine their binding activity, double-stranded DNA-based and PDP-based decoy molecules were incubated with different amounts of microparticles in the presence of 100 ng of either P-32-labeled DNA-DNA or DNA-PDP hybrid molecules or cold PDP-PDP hybrids. The complexes were analyzed by agarose gel electrophoresis. The resistance of P-32-labeled DNA-DNA and DNA-PDP molecules in the presence of serum or cellular extracts was evaluated after binding to CM by gel electrophoresis analysis. DDAB18 Eudragit RS 100 microspheres are able to bind to DNA-PDP and PDP-PDP hybrids, to deliver these molecules to target cells and to protect DNA-PDP molecules from enzymatic degradation in simulated biological fluids. In addition, when assayed in ex vivo conditions, DDAB18 Eudragit RS 100 microspheres exhibited low toxicity. The results presented in this paper demonstrate that CM can be considered suitable formulations for pharmacogenomic therapy employing double-stranded PDP chimeras. Copyright (C) 2004 National Science Council, ROC and S. Karger AG, Basel.
Complexation to cationic microspheres of double-stranded peptide nucleic acid-DNA chimeras exhibiting decoy activity