Non-viral delivery of nucleic acids and image-guided assessment of in vivo performance of multifunctional nanomedicines [Elektronische Ressource] / vorgelegt von Olivia Monika Merkel
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2009
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278
pages
Ebook
2009
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Publié par
Publié le
01 janvier 2009
Nombre de lectures
50
Poids de l'ouvrage
10 Mo
Publié par
Publié le
01 janvier 2009
Nombre de lectures
50
Poids de l'ouvrage
10 Mo
Non-Viral Delivery of Nucleic Acids and
Image-Guided Assessment of in vivo
Performance of Multifunctional
Nanomedicines
Dissertation
zur Erlangung des Doktorgrades der Naturwissenschaften (Dr. rer. nat.)
dem
Fachbereich Pharmazie der Philipps-Universität Marburg
vorgelegt von
Olivia Monika Merkel
aus Villingen-Schwenningen
Marburg/Lahn 2009
Vom Fachbereich Pharmazie der Philipps-Universität Marburg als Dissertation am
24.08.2009 angenommen.
Erstgutachter: Prof. Dr. Thomas Kissel
Zweitgutachter: Prof. Dr. Carsten Culmsee
Drittgutachter: Prof. Dr. Werner Seeger
Tag der mündlichen Prüfung: 25.08.2009
Die vorliegende Arbeit entstand auf Anregung und unter Leitung von
Herrn Prof. Dr. Thomas Kissel
am Institut für Pharmazeutische Technologie und Biopharmazie
der Philipps-Universität Marburg.
Gewidmet meinen Eltern
in Liebe und Dankbarkeit
TABLE OF CONTENTS
1 INTRODUCTION ............................................................................................... 1
1.1 Nanomedicine ............................................................................................................. 2
1.2 Non-Viral Delivery of Nucleic Acids........................................................................ 4
1.3 Targeted Gene Delivery Using Cell Specific Ligands............................................ 7
1.4 Dendrimers in Gene Delivery ................................................................................... 8
1.5 Concepts of siRNA Mediated Post-Transcriptional Gene Silencing ................. 11
1.6 Pulmonary siRNA Delivery..................................................................................... 15
1.7 Nuclear Imaging of siRNA Delivery ...................................................................... 17
1.8 Polymer Genomics and Nanotoxicology................................................................ 20
1.9 Structure of the Thesis: Aims and Outline ........................................................... 22
1.10 References ................................................................................................................. 24
PART I: NON-VIRAL IN VITRO
DELIVERY OF pDNA
2 INTEGRIN ALPHA-ν-BETA-3 TARGETED GENE DELIVERY USING RGD
PEPTIDOMIMETIC CONJUGATES WITH COPOLYMERS OF PEGYLATED
POLY(ETHYLENE IMINE) ..................................................................................... 44
2.1 Abstract ..................................................................................................................... 45
2.2 Introduction .............................................................................................................. 45
2.3 Materials and Methods............................................................................................ 46
2.4 Results and Discussion............................................................................................. 52
i
2.5 Conclusion................................................................................................................. 70
2.6 Acknowledgements................................................................................................... 70
2.7 References ................................................................................................................. 71
3 POLYCATIONIC TRIAZINE-BASED DENDRIMERS: EFFECT OF
PERIPHERAL GROUPS ON TRANSFECTION EFFICIENCY ............................. 77
3.1 Abstract ..................................................................................................................... 78
3.2 Introduction .............................................................................................................. 78
3.3 Results and Discussion............................................................................................. 80
3.4 Conclusion................................................................................................................. 87
3.5 Experimental............................................................................................................. 88
3.6 Acknowledgements................................................................................................... 95
3.7 Notes and references ................................................................................................ 95
4 TRIAZINE DENDRIMERS AS NON-VIRAL GENE DELIVERY SYSTEMS:
EFFECTS OF MOLECULAR STRUCTURE ON BIOLOGICAL ACTIVITY.......... 99
4.1 Abstract ................................................................................................................... 100
4.2 Introduction ............................................................................................................ 100
4.3 Materials and Methods.......................................................................................... 101
4.4 Results and Discussion........................................................................................... 105
4.5 Conclusion............................................................................................................... 118
4.6 Acknowledgements................................................................................................. 119
4.7 References ............................................................................................................... 119
ii
PART II: NON-VIRAL IN VITRO AND
IN VIVO DELIVERY OF siRNA
5 IN VIVO SPECT AND REAL-TIME GAMMA CAMERA IMAGING OF
BIODISTRIBUTION AND PHARMACOKINETICS OF SIRNA DELIVERY USING
AN OPTIMIZED RADIOLABELING AND PURIFICATION PROCEDURE......... 125
5.1 Abstract ................................................................................................................... 126
5.2 Introduction ............................................................................................................ 126
5.3 Materials and Methods.......................................................................................... 128
5.4 Results and Discussion........................................................................................... 130
5.5 Conclusion............................................................................................................... 141
5.6 Acknowledgment.................................................................................................... 142
5.7 Supporting Information ........................................................................................ 142
5.8 References ............................................................................................................... 142
6 STABILITY OF SIRNA POLYPLEXES FROM POLY(ETHYLENIMINE) AND
POLY(ETHYLENIMINE)-G-POLY(ETHYLENE GLYCOL) UNDER IN VIVO
CONDITIONS: EFFECTS ON PHARMACOKINETICS AND BIODISTRIBUTION
MEASURED BY FLUORESCENCE FLUCTUATION SPECTROSCOPY AND
SINGLE PHOTON EMISSION COMPUTED TOMOGRAPHY (SPECT) IMAGING
148
6.1 Abstract ................................................................................................................... 149
6.2 Introduction ............................................................................................................ 149
6.3 Materials and Methods.......................................................................................... 151
6.4 Results and Discussion........................................................................................... 157
6.5 Conclusion............................................................................................................... 177
ii i
6.6 Acknowledgements................................................................................................. 178
6.7 References ............................................................................................................... 178
7 NON-VIRAL SIRNA DELIVERY TO THE LUNG – INVESTIGATION OF PEG-
PEI POLYPLEXES AND THEIR IN VIVO PERFORMANCE .............................. 184
7.1 Abstract ................................................................................................................... 185
7.2 Introduction ............................................................................................................ 185
7.3 Experimental Section............................................................................................. 187
7.4 Results and Discussion........................................................................................... 192
7.5 Conclusion............................................................................................................... 212
7.6 Acknowledgements................................................................................................. 213
7.7 References ............................................................................................................... 213
8 OFF-TARGET EFFECTS IN NON-VIRAL SIRNA DELIVERY – A STUDY ON
THE EFFECT OF POLYMER GENOMICS ON IN VITRO CELL CULTURE
MODELS....................................................................................................
