The development of sustained release formulation for pharmaceutical proteins based on vesicular phospholipid gels [Elektronische Ressource] / vorgelegt von Weiwei Tian

Weiwei Tian

Informations

• Publié par: ludwig-maximilians-universitat_munchen
• Publié le: 01 janvier 2010
• Nombre de lectures: 14
• Langue: English
• Poids de l'ouvrage: 3 Mo

Extrait

The Development of Sustained Release Formulation
for Pharmaceutical Proteins based
on Vesicular Phospholipid Gels




Dissertation



zur Erlangung des Doktorgrades der
Fakultät für Chemie und Pharmazie der
Ludwig-Maximilians-Universität München





vorgelegt von
Weiwei Tian
aus Tianjin, China

München 2010 Dissertation zur Erlangung des
Doktorgrades
der Fakultät für Chemie und Pharmazie
der Ludwig-Maximilians-Universität
München




The Development of Sustained Release Formulation
for Pharmaceutical Proteins based
on Vesicular Phospholipid Gels





Weiwei Tian

aus Tianjin, China




München 2010





Erklärung

Diese Dissertation wurde im Sinne von § 13 Abs. 3 bzw. 4 der Promotionsordnung
vom 29. Januar 1998 von Herrn Prof. Dr. G. Winter betreut.

Ehrenwörtliche Versicherung

Diese Dissertation wurde selbständig, ohne unerlaubte Hilfe erarbeitet.




München, den 01.12.2009







……………………………………
Weiwi Tan





Dissertation eingereicht am: 02.02.2010

1. Gutachter: Prof. Dr. G. Winter
2. Gutachter: Prof. Dr. M. Brandl

Mündliche Prüfung am: 24.02.2010 ACKNOWLEDGEMENTS

The present work was carried out from Oct. 2006 to Dec. 2009 under the supervision
of Prof. Dr. Gerhard Winter at the Department of Pharmacy, Pharmaceutical
Technology and Biopharmaceutics, Ludwig-Maximilians-University, Munich.

Foremost, I would express my deepest gratitude to my supervisor, Prof. Dr. Gerhard
Winter offering me the possibility to join his research group. I am extremely grateful
for his inspiring encouragement, professional guidance and scientific support.
Especially, I want to thank him for giving me chances to present the work in the
international conferences.

I am also deeply grateful to Prof. Dr. Martin Brandl, my second supervisor, who
introduced me the technological education and granted scientific advices and
solutions. Thanks for his help I could always rely on during the last years.

Especially, I would like to thank the Chinese Scholarship Counsel for the financial
support.

Moreover, I especially thank Dr. Sandra Schulze for the guidance over the second
year.

Many thanks to all the past and present colleagues in the research group of Prof. Dr.
Winter and Prof. Dr. Frieß for creating an outstanding working climate. I would
especially thank for the relief they have granted me at the beginning of my study.

Furthermore, I am indebted to Dr. Matthias Hadesbeck, Ingrid Schmidt, Monique-
Claudine Esnouf from the International Affairs of the university for supporting me so
much with solving the problems outside the laboratory.
.
I would like to acknowledge Dr. Stefan Buchmann, from the Sport orthopaedic, TUM,
Germany, for performing the animal experiments. Thanks are extended to his clinic
research team. Thanks are extended to the University of Tromso, Norway for offering the dual
asymmetric centrifuge as well as to Lipoid GmbH, Ludwigshafen, Germany and
PHOSPHOLIPID GmbH, Cologne, Germany, for providing various lipids.

Thanks are also extended to Prof. Dr. W. Frieß, Prof. Dr. F. Boeckler, and Prof. Dr. F.
Bracher for serving as members of my thesis advisor committee.

Finally I would like to thank my parents and my friend Shen for their enduring love
and constant encouragement. Thank you very much!
TABLE OF CONTENT
Chapter I: Introduction............................................................................................. 1
1. General consideration on pharmaceutical proteins......................................................2
2. Administration routes and existing problems ...............................................................3
2.1. Parenteral delivery .......................................................................................................4
2.2. Oral delivery.................................................................................................................4
2.3. Alternative routes.........................................................................................................6
2.4. Controlled drug delivery system...................................................................................7
3. Polymeric system .............................................................................................................8
3.1. Poly (Lactic-co-glycolic acid) system ...........................................................................9
3.2. Hydrogel.....................................................................................................................10
4. Lipidic systems...............................................................................................................11
4.1. Lipid implants12
4.2. Multivesicular liposomes ............................................................................................14
4.2.1. Preparation method................................................................................................14
4.2.2. Influence factors on the release .............................................................................15
4.2.3. Mvls for controlled drug delivery............................................................................17
4.3. Vesicular phospholipid gels .......................................................................................21
4.3.1. Preparation methods..............................................................................................21
4.3.2. Characterization of vpgs.........................................................................................24
4.3.3. Application of vpgs for drug delivery ......................................................................26
Chapter II: Aim of the thesis.................................................................................. 30
Chapter III: Materials and methods ....................................................................... 32
1. Materials ..........................................................................................................................32
1.1. Fluorescein isothiocyanate-dextran (FITC-dextran)...................................................32
1.2. Erythropoetin (EPO)...................................................................................................32
1.3. Granulocyte colony-stimulating factor (G-CSF) .........................................................33
1.4. Monoclonal antibody immunoglobulin G (mAb)33
1.5. Phospholipids.............................................................................................................34
1.6. Chemicals and reagents ...........................................................................................34
2. Methods ...........................................................................................................................35
2.1. Up-Concentration of proteins .....................................................................................35
2.2. Preparation of vesicular phospholipid gels ................................................................35
2.2.1. Preparation by high pressure homogenization.......................................................35
2.2.2. Preparation by dual asymmetric centrifugation ......................................................36
2.3. Microscopic studies....................................................................................................37
2.4. Rheology....................................................................................................................37 2.5. Texture analysis.........................................................................................................37
2.6. Dispersions of VPGs..................................................................................................37
2.7. Particle size analysis and zeta potential measurement .............................................37
2.8. Turbidity .....................................................................................................................38
2.9. Extraction of proteins from the VPG matrix................................................................38
2.9.1. Extraction of EPO...................................................................................................38
2.9.2. Extraction of G-CSF ...............................................................................................38
2.9.3. Extraction of mAb IgG ............................................................................................39
2.10. Sodium dodecyl sulphate polyacrylamide gel electrophoresis...................................39
2.11. In vitro release tests39
2.12. Quantification of released drugs ................................................................................40
2.12.1. Determination of fitc-dextran concentration by fluoresc