Design, synthesis and characterization of novel exogenous smart, bioresponsive contrast agents for magnetic resonance and optical imaging [Elektronische Ressource] = Design, Synthese und Charakterisierung neuartiger exogener intelligenter, bioreaktiver Kontrastmittel für die Magnetresonanz und optische Bildgebung / vorgelegt von Anurag Mishra

Anuragrk

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• Publié par: eberhard_karls_universitat_tubingen
• Publié le: 01 janvier 2008
• Nombre de lectures: 55
• Langue: English
• Poids de l'ouvrage: 2 Mo

Extrait

Design, Synthesis and Characterization of Novel
Exogenous Smart/Bioresponsive Contrast Agents
for Magnetic Resonance and Optical Imaging

Design, Synthese und Charakterisierung neuartiger
exogener intelligenter/bioreaktiver Kontrastmittel
für die Magnetresonanz und optische Bildgebung



DISSERTATION



der Fakultät für Chemie und Pharmazie
der Eberhard-Karls-Universität Tübingen
zur Erlangung des Grades eines Doktors
der Naturwissenschaften



2008


vorgelegt von

Anurag Mishra










































Tag der mündlichen Prüfung: 19. Dezember 2007


Dekan: Prof. Dr. Lars Wesemann


1. Berichterstatter Prof. Dr. Klaus Albert

2. Berichterstatter Prof. Dr. Martin E. Maier
This PhD thesis was prepared at the Max-Planck Institute for Biological Cybernetics
(chemistry laboratory of the Department Physiology of Cognitive Processes, Prof. Dr.
Nikos K. Logothetis) in collaboration with the Institute for Organic Chemistry,
Eberhard-Karls-University, Tübingen under the guidance of Prof. Dr. Klaus Albert
during the period from December 2003 to December 2007.



















Hereby I declare the fact that I am writing this work and no different than the
indicated aids have been used.


Tübingen, December, 2007Abbreviations
Abbreviations
Boc tert. Butoxy carbonyl
tBu tert butyl
c Concentration
δ Chemical shift in ppm (NMR)
CBz Chlorobenzoformate
CCl Carbon tetrachloride 4
CDCl Chloroform-deuterated 3
CH Cl Dichlormethane 2 2
CLIO Cross-linked iron oxid
DCC Dicyclohexylcarbodiimide
DMAP 4-Dimethylaminopyridine
DMF N,N-Dimethylformamide
DO3AMP DO3A-methyl phosphonic acid
DO Deuteriumoxid 2
EDC 1-Ethyl-3-(3’-dimethylaminopropyl) carbodiimide
EDTA Ethylenediaminetetraacetic acid
EGTA Ethylene glycol bis(2-aminoethylether)-N,N,N’,N’-
tetraacetic acid
Equ Equation
ESI-MS Electron Spray Ionization-Mass Spectrometry
Et Ethyl
EtO Diethyl ether 2
EtOAc Ethyl acetate
EuCl .6H O Europium trichloride hexahydrate 3 2
Fig. Figure
FITC Fluorescein isothiocyanate
g gram (s)
GABA gamma-amino butyric acid
GdCl .6HO Gadolinium trichloride hexahydrate 3 2
h hour (s)
H Hydrogen 2Abbreviations
HBr Hydrobromic acid
HCl drochloric acid
HOBt Hydroxybenzotriazole
HPLC high performance liquid chromatography
HSA human serum albumin
ESI-HRMS Electron spray ionization-high resolution mass
spectrometry
Hz hertz
i-Pr isopropyl
J coupling constant (NMR)
K CO Potassiumcarbonate2 3
λ lambda
LiOH Lithium hydroxide
Ln lanthanide
Me Methyl
MeCN acetonitrile
MeOHthanol
MeODthanol-deuterated
mg milligram
m/z Mass to charge ratio (MS)
mL milliliter (s)
mmol millimole
mMillimolar
MoAb Monoclonal antibody
Na CO Sodiumcarbonate 2 3
NaHCOodium bi3
NaOHodiumhydroxide
Na SO Sodiumsulphate 2 4
NMM N-Methylmorpholine
NH/MeOH methanolic ammonia 3
p para
Pd-C Palladiumcarbon
PPH triphenylphosphine 3
Ph Phenyl Abbreviations
PP Pyridoxyl-5-phosphate 5
PyBroP Bromo-tris-pyrrolidino-phosphonium
hexafluorophosphate
Ra-Ni Raney nickle
R Retention factor (TLC) f
RT Retention time (HPLC)
RP-HPLC Reverse phase- high performance liquid
chromatography
SPIO superparamagnetic iron oxide
TEA triethylamine
TFA Trifluroacetic acid
THF Tetrahydrofuran
UV Ultraviolet






Acknowledgements

Acknowledgements

Knowledge in itself is a continuous process. At this moment of my substantial
enhancement, I have found this rare opportunity to evince a word of thanks to all
those who have played a key role in the successful completion of my PhD thesis.
My heartfelt thanks to Prof. Dr. Klaus Albert, Institute for Organic Chemistry,
Tübingen University, for providing his unconditional support, guidance and optimal
resources allocation that helped me in the successful completion of my doctoral
projects.
I wish to express my deepest reverence to Prof. Dr. Nikos K. Logothetis,
Director, Department Physiology of Cognitive Processes, Max Planck Institute for
Biological Cybernetics, for his unrestricted support, suggestions, valuable guidance
and providing me the facilities during the course of the investigation.
I owe a sincere thanks to my esteemed advisors Dr. Josef Pfeuffer, Head of
MR Imaging and Spectroscopy, Max Planck Institute for Biological Cybernetics
(currently Siemens AG, Medical Solutions, Boston, USA) and Dr. Anil Mishra,
INMAS, Delhi, India for their sincere guidance, co-operation, constant
encouragement, patience and belief in me.
I would like to thank Prof. Dr. Martin E. Maier, Institute for Organic
Chemistry, Tübingen University, for his helpful reviewing the doctoral thesis and
giving valuable comments and suggestions.
I am very thankful to Prof. Dr. Almut Schuez and Dr. Santiago Canals for
their help in performing and designing in vivo histological experiments of
neuroanatomical tracers and their advices in writing of the some biological part of
thesis.
I express my special thanks to our biology group, MPI for Biological
Cybernetics, Dept. Ugurbil: Dr. Joern Engelmann, my lovely wife Ritu Mishra and
Hildegard Schulz for performing cell studies and tremendous guidance. The work in
this thesis would not have been possible without their collaboration.
I express my cordial gratitude to Dipl.-Ing. Michael Beyerlein and Dr. Rolf
Pohmann for performing the MR experiments at 1.5T, 3T and 7T. Acknowledgements

I also thank Dr. Eva Jakab Toth, Dr. Petra Fouskova, Centre de Biophysique
17Moléculaire, CNRS, Orleans, France, for performing NMRD, O-NMR and HR-
UVis measurements of some of the calcium sensitive contrast agents and Prof. Dr.
David Parker, Department of Chemistry, Durham University, Durham, U.K. for
luminescence studies of pH sensitive contrast agents.
I am thankful to the all members I worked with in the group of Prof. Dr. Klaus
Albert and Prof. Nikos K. Logothetis for their timely help and support.
Thanks to all members of our bioconjugate group: Dr. Ilgar Mammadov, Kirti
Dhingra, Deepti Jha, Aneta Brud, Dr. Wu Su, Dr. Xiao-zhe Zhang and Dr. Goran
Angelovski for your kind support and friendly atmosphere during my work. Thanks to
Conchy Moya and Rainer Hirt for all the help on the administrative arrangements.
Many good friends of mine, such as Suryadeep Dash, Vikram Alva, Dr.
Srinivasa Mariamganti, Deepti Jha, Kirti Dhingra, Rajendar Joshi and others have
enriched my academic and social activity in Tübingen. I will never forget your help,
optimism and energy.
Finally, I would like to pay my tribute to the constant support of my Parents
and brother their love, understanding and encouragement throughout my life. I don’t
have words to explain my wife Ritu and my daughter Arshiya. I could say without
them I might have not finished my PhD in time.
I also thank all those who could not find a separate name but have helped
directly and indirectly.














Table of Contents


Table of Contents

1. CHAPTER 1. Literature Review………………………………………………..…1
1.1. Introduction....………………………………………..…………………..….2
1.1.1. Advantages and Disadvantages of MR Techniques over non MR
Techniques…...……………......................................................................2
1.1.2. Associated issues when working in a High-Magnetic-Field
Environments…………………………………………….….……..…….4
1.2. Background and Significance………..………………………….……..........5
1.2.1. Relaxation Parameters………………………………….………….…..5
1.2.2. Endogenous and Exogenous Contrast Agents………………….……..7
1.2.3. Lanthanide Metal Chelates…………………………………………....7
1.2.4. Evolution of Contrast Agents………………………………………....8
1.2.5. Smart/Bioactivated Contrast Agents as Functional Biochemical
Markers………………………………………………………………....11
1.2.5.1. pH-Activated Contrast Agent……………..…….……………11
1.2.5.2. Metal Ion Concentration Activated Contrast Agents……...…13
1.2.5.3. Enzyme-Activated Contrast Agents.….………..……………..14
1.3. Project Conceptulations……………………………………………………15
1.3.1. Objective…………………………………………..…15
1.3.2. Specific Aim…………………..……………………………………...15
2. CHAPTER 2. New Class of Gd-based Targeted Contrast Agents for MR and
Optical Imaging Derived from DO3A-ethylamine………………...…………….16
2.1. Introduction……………………………………………………………...…17
2.2. Experimental Section………………………………………………………19
2.2.1. In v