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2003
Le téléchargement nécessite un accès à la bibliothèque YouScribe Tout savoir sur nos offres
191
pages
English
Documents
2003
Le téléchargement nécessite un accès à la bibliothèque YouScribe Tout savoir sur nos offres
Publié par
Publié le
01 janvier 2003
Nombre de lectures
42
Langue
English
Poids de l'ouvrage
4 Mo
Publié par
Publié le
01 janvier 2003
Langue
English
Poids de l'ouvrage
4 Mo
Biotechnologie-Gesellschaft
Mittelhessen mbH
HESylation – a new technology for polymer conjugation to
biologically active molecules
Modification of proteins and low molecular weight
substances with hydroxyethyl starch (HES)
Inauguraldissertation
Michele Orlando – Gießen, 2003
HESylation – a new technology for polymer conjugation to
biologically active molecules
Modification of proteins and low molecular weight
substances with hydroxyethyl starch (HES)
INAUGURALDISSERTATION
zur Erlangung des Grades
Doktor der Naturwissenschaften
- Dr. rer. nat. –
des Fachbereichs Biologie, Chemie und Geowissenschaften, FB08
der Justus Liebig Universität Giessen
vorgelegt von
Michele Orlando
Geboren am 12. Dezember 1975 in Potenza (Italien)
Gießen, 2003
Alla mia famiglia. The experiments for the present work were performed from September 2000 until July 2003
in the laboratories of the Biotechnologie-Gesellschaft Mittelhessen mbH (Bim GmbH) in
Giessen under the supervision of Prof. Dr. Alfred M. Pingoud.
Dekan: Prof. Dr. Jürgen Mayer
Institut für Biologiedidaktik, FB 08
der Justus Liebig Universität Gießen
Karl-Glöckner-Straße 21c
D-35394 Gießen
Referent: Prof. Dr. Alfred M. Pingoud
Institut für Biologie, Chemie und Geowissenschaften, FB 08
der Justus Liebig Universität Gießen
Heinrich-Buff-Ring 58
D-35392 Gießen
Korreferent: Prof. Dr. Jürgen Hemberger
Fachbereich KMUB - Biotechnologie
der Fachhochschule Gießen-Friedberg
Wiesenstraße 14
D-35390 Gießen
"L’alto disio che mo t’infiamma e urge,
d’aver notizia di ciò che tu vei,
tanto mi piace più quanto più turge;
ma di quest’acqua convien che tu bei
prima che tanta sete in te si sazi"
(Dante Alighieri – Paradiso XXX, 70-74) Thanks to...
… everyone who stood by me (even if they were distant) during these years of hard
work in the conclusion of my Ph. D. thesis.
… BIM GmbH, especially to Professor J. Hemberger and Dr. J. Bille (or simply Jürgen
and Achim) for having given me the opportunity to carry out this research and to use the
results for my thesis.
… the staff of BIM (including those who don’t belong to the company anymore):
Jürgen, Achim, Alex, Heiko, Antje F., Kai, Antje K., Dirk, Uwe, Claudia, Dagmar,
Manuela, Jeanne, Eva, Andreas, Jean-Charles, Nicolas, Josè, Paolo, Valentino either for
their scientific contribution to the production of the results, or for their friendship and
support.
… Professor Dr. A. Pingoud for having “adopted” me when I was searching for a
“Doktorvater” and for the endless patience, cordiality and availability that he always
demonstrated.
… Dr. P. Czodrowsky for the advices on molecular modelling.
… my family that, despite the distance, has always given me support, strength and
motivation. Thanks for their worries, for their thoughts and for the priceless sacrifices
that they had to face. Thanks for all the love and affection as well as all the phone calls!
… all my friends for the good moments we had together and for having compensated
with their warmth the severe climate of this country.
IEidesstattliche Erklärung
Hiermit versichere ich, dass ich die vorliegende Arbeit selbständig angefertigt
habe und keine anderen als die angegebenen Hilfsmittel verwendet habe. Die
Arbeit wurde so oder in ähnlicher Form noch keinem anderen Prüfungsausschuss
vorgelegt.
Gießen, 9. Oktober 2003
Michele Orlando
IICurriculum vitae
Born: 12. December 1975
in: Potenza (Italy)
Education:
1989-1994 High school at Liceo Classico “Q. O. Flacco” Potenza (Italy) graduated
with the note 60/60.
1994-1999 Study of Pharmaceutical Chemistry and Technology (CTF) at the
“Federico II” University in Naples (Italy).
1997-1999 24 months of laboratory works in the department of Pharmaceutical
Chemistry in collaboration with prof. G. Caliendo reseach group. Main
topics of the researches were organic synthesis of pharmacologically
active molecules and peptide synthesis.
23.07.1999 Master degree obtained discussing the thesis: “Synthesis and
characterisation of new 1,2,3-benzotriazin-4-one-arylpiperazine
derivatives as 5-HT serotonin receptor ligands”. Final note 110/110 1A
summa cum laude.
since 05.2001 Ph.D. student in biochemistry at the “Justus Liebig” University of
Giessen (Germany). The research works are performed in the laboratories
of Biotechnologie Gesellshaft Mittelhessen mbH under the supervision of
prof. dr. A. Pingoud.
Professional experience:
09.1999 – 12.1999 Training period at Biotechnologie Gesellschaft Mittehessen mbH
(Giessen, Germany) supported by the Leonardo EU program.
01.2000 – today Working at Biotechnologie Gesellshaft Mittelhessen mbH as researcher.
III Curriculum vitae
---------------------------------------------------------------------------------------------------------------------
Patent applications:
03.2002 Kopplung von Proteinen an ein modifiziertes Polysaccharid (Coupling of
proteins to a modified polysaccharide)
International publication N°: WO03074087
03.2002 Kopplung niedermolecularer Substanzen an ein modifiziertes
Polysaccharid (Coupling of low molecular weight substances to a
modified polysaccharide).
International publication N°: WO03074088
11.2002 Water-soluble prodrugs of Propofol.
PCT N°: PCT18421
03.2003 Pharmaceutically active oligosaccharide conjugates.
European patent application N°: EP27268
Publications:
2003 Highly water soluble derivative of Amphotericin B having the same
antimycotic potential of the original drug. (in preparation)
2003 Increase in selectivity of Amphotericin B after conjugation with a
biocompatible polysaccharide. (in preparation)
IVIndex
Page
THANKS TO... I
EIDESTATTLICHE ERKLÄRUNG II
CURRICULUM VITAE III
INDEX V
INDEX OF THE FIGURES IX
INDEX OF THE TABLES XIII
INDEX OF THE ABBREV IATIONS XIV
ZUSAMMENFASSUNG XVII
ABSTRACT XIX
Chapter 1: INTRODUCTION 1
1.1 Polymer protein conjugation 3
1.1.1 Dextran-protein conjugates 4
1.1.2Polyethylene glycol-proteinconjugates 5
1.2 Benefits of polymer modification for protein pharmaceuticals 5
1.2.1 Increasd plasm half-ie 5
1.2.2 Reducerenalclearance 7
1.2.3 d cellular clearance 7
1.2.4 Reduced proteolysis 8
1.2.5 Reducedimmunogenicity and antigenicity 10
1.2.6 Increased solubility 12
1.3 Problems and solutions in polymer-protein conjugation 13
1.4 Polymer modification of small drugs 15
1.5 Advantages in preparation of bioconjugates with low molecular weight drugs 17
1.6 Objectives of the present work 18
Chapter 2: HYDROXYETHYL STARCH 20
2.1 Structural unit 20
2.1.1 Amylose 21 1.2Amylopectin
2.2 Preparation of hydroxyethyl starch 23
V