caa a22 4500 445313463 CHVBK 20180317142638.0 cr unu---uuuuu 170323e20110401xx s 000 0 eng 10.1007/s11095-010-0225-4 doi (NATIONALLICENCE)springer-10.1007/s11095-010-0225-4 Poly(I:C)-Mediated Tumor Growth Suppression in EGF-Receptor Overexpressing Tumors Using EGF-Polyethylene Glycol-Linear Polyethylenimine as Carrier [Elektronische Daten] [David Schaffert, Melinda Kiss, Wolfgang Rödl, Alexei Shir, Alexander Levitzki, Manfred Ogris, Ernst Wagner] ABSTRACT: Purpose: To develop a novel polyethylenimine (PEI)-based polymeric carrier for tumor-targeted delivery of cytotoxic double-stranded RNA polyinosinic:polycytidylic acid, poly(I:C). The novel carrier should be chemically less complex but at least as effective as a previously developed tetra-conjugate containing epidermal growth factor (EGF) as targeting ligand, polyethylene glycol (PEG) as shielding spacer, 25kDa branched PEI as RNA binding and endosomal buffering agent, and melittin as endosomal escape agent. Methods: Novel conjugates were designed employing a simplified synthetic strategy based on 22kDa linear polyethylenimine (LPEI), PEG spacers, and recombinant EGF. The efficacy of various conjugates (different PEG spacers, with and without targeting EGF) in poly(I:C)-mediated cell killing was evaluated in vitro using two human U87MG glioma cell lines. The most effective polyplex was tested for in vivo activity in A431 tumor xenografts. Results: Targeting conjugate LPEI-PEG2kDa-EGF was found as most effective in poly(I:C)-triggered killing of tumor cells in vitro. The efficacy correlated with glioma cell EGFR levels. Repeated intravenous administration of poly(I:C) polypexes strongly retarded growth of A431 human tumor xenograft in mice. Conclusions: The optimized LPEI-PEG2kDa-EGF conjugate displays reduced chemical complexity and efficient poly(I:C)-mediated killing of EGFR overexpressing tumors in vitro and in vivo. Springer Science+Business Media, LLC, 2010 epidermal growth factor nationallicence polyplex nationallicence receptor-mediated delivery nationallicence RNA delivery nationallicence tumor targeting nationallicence HBG : Hepes buffered glucose (5% (w/v) glucose, 20mM Hepes, pH7.4) nationallicence HEPES : N-2-hydroxyethylpiperazine-N′-2-ethane sulfonic acid nationallicence IEX : ion-exchange chromatography nationallicence LPEI : linear polyethylenimine with an average molecular weight of 22kDa nationallicence OPSS : orthopyridyl dithio nationallicence PEG : polyethylene glycole nationallicence p(I) : poly inosinic acid nationallicence poly(Glu) : polyglutamic acid nationallicence poly(I:C) : poly inosinic-cytidylic acid nationallicence SEC : size exclusion chromatography nationallicence Schaffert David Pharmaceutical Biotechnology, Department of Pharmacy Center for Drug Research, Ludwig-Maximilians-Universität Munich, Butenandtstr. 5-13, 81377, Munich, Germany aut Kiss Melinda Pharmaceutical Biotechnology, Department of Pharmacy Center for Drug Research, Ludwig-Maximilians-Universität Munich, Butenandtstr. 5-13, 81377, Munich, Germany aut Rödl Wolfgang Pharmaceutical Biotechnology, Department of Pharmacy Center for Drug Research, Ludwig-Maximilians-Universität Munich, Butenandtstr. 5-13, 81377, Munich, Germany aut Shir Alexei Unit of Cellular Signaling, Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel aut Levitzki Alexander Unit of Cellular Signaling, Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel aut Ogris Manfred Pharmaceutical Biotechnology, Department of Pharmacy Center for Drug Research, Ludwig-Maximilians-Universität Munich, Butenandtstr. 5-13, 81377, Munich, Germany aut Wagner Ernst Pharmaceutical Biotechnology, Department of Pharmacy Center for Drug Research, Ludwig-Maximilians-Universität Munich, Butenandtstr. 5-13, 81377, Munich, Germany aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/4(2011-04-01), 731-741 0724-8741 28:4<731 2011 28 11095 https://doi.org/10.1007/s11095-010-0225-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-010-0225-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Schaffert David Pharmaceutical Biotechnology, Department of Pharmacy Center for Drug Research, Ludwig-Maximilians-Universität Munich, Butenandtstr. 5-13, 81377, Munich, Germany aut NATIONALLICENCE 700 1- Kiss Melinda Pharmaceutical Biotechnology, Department of Pharmacy Center for Drug Research, Ludwig-Maximilians-Universität Munich, Butenandtstr. 5-13, 81377, Munich, Germany aut NATIONALLICENCE 700 1- Rödl Wolfgang Pharmaceutical Biotechnology, Department of Pharmacy Center for Drug Research, Ludwig-Maximilians-Universität Munich, Butenandtstr. 5-13, 81377, Munich, Germany aut NATIONALLICENCE 700 1- Shir Alexei Unit of Cellular Signaling, Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel aut NATIONALLICENCE 700 1- Levitzki Alexander Unit of Cellular Signaling, Department of Biological Chemistry, The Hebrew University of Jerusalem, Jerusalem, Israel aut NATIONALLICENCE 700 1- Ogris Manfred Pharmaceutical Biotechnology, Department of Pharmacy Center for Drug Research, Ludwig-Maximilians-Universität Munich, Butenandtstr. 5-13, 81377, Munich, Germany aut NATIONALLICENCE 700 1- Wagner Ernst Pharmaceutical Biotechnology, Department of Pharmacy Center for Drug Research, Ludwig-Maximilians-Universität Munich, Butenandtstr. 5-13, 81377, Munich, Germany aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/4(2011-04-01), 731-741 0724-8741 28:4<731 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer