Non-viral delivery of nucleic acids and image-guided assessment of in vivo performance of multifunctional nanomedicines [Elektronische Ressource] / vorgelegt von Olivia Monika Merkel

Olivia Merkel

Sujets

• Naturwissenschaften

Informations

• Publié par: philipps-universitat_marburg
• Publié le: 01 janvier 2009
• Nombre de lectures: 50
• Poids de l'ouvrage: 10 Mo

Extrait

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....................................................................................................