Dimeric ligands for GPCRs involved in human reproduction: synthesis and biological evaluation
Bonger, K.M.
Citation
Bonger, K. M. (2008, December 19). Dimeric ligands for GPCRs involved in human reproduction: synthesis and biological evaluation. Retrieved from
https://hdl.handle.net/1887/13368
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Dimeric ligands for GPCRs involved in human reproduction:
synthesis and biological evaluation
PROEFSCHRIFT
ter verkrijging van
de graad van Doctor aan de Universiteit Leiden,
op gezag van Rector Magnificus prof.mr. P.F. van der Heijden, volgens besluit van het College voor Promoties
te verdedigen op vrijdag 19 december 2008 klokke 12.30 uur
door
Kimberly Michelle Bonger
geboren te Toronto, Canada in 1980
Promotiecommissie
Promotores : Prof. Dr. H. S. Overkleeft : Prof. Dr. G. A. van der Marel
Co-promotor : Dr. C. M. Timmers (Schering-Plough research institute)
Overige leden : Dr. R. J. B. H. N. van den Berg
Prof. Dr. A. P. IJzerman
Dr. C. J. van Koppen (Schering-Plough research institute)
Prof. Dr. J. Lugtenburg
Prof. Dr. R. V. A. Orru (vrije Universiteit amsterdam)
This research was carried out in collaboration with Schering-Plough Research Institute (Oss), as part of the NWO-Combichem program. The project has been financially supported by the Council for Chemical Sciences of the Netherlands Organization for Scientific Research (CW- NWO) and the Schering-Plough Research Institute (Oss).
Cover: Topograph obtained by atomic-force microscopy showing the regular rearrangement of rhodopsin in the native disc membrane. Picture adapted from Fotiadis, D.; Liang, Y.;
Filipek, S.; Saperstein, D. A.; Engel, A.; Palczewski, K. Nature 2003, 421, 127-128.
Table of Contents
List of Abbreviations 6
Chapter 1 9
General introduction
Chapter 2 37
Synthesis and evaluation of homodimeric GnRHR ligands
Chapter 3 59
Synthesis and evaluation of homodimeric GnRHR antagonists having a rigid propargylated benzene core
Chapter 4 83
GnRHR binding and antagonism of dimeric systems appear dependent on the nature of the parent pharmacophore
Chapter 5 101
Discovery of selective LHR agonists by the bivalent ligand method
Chapter 6 119
Hydroxylated prolines as spacers for dimeric LHR agonists
Chapter 7 139 Oligoproline helix as a scaffold for potent, selective and
structurally defined dimeric ligands for the LHR
Chapter 8 159
Synthesis and pharmacological evaluation of dimeric FSHR antagonists
Chapter 9 183
Synthesis and pharmacological evaluation of heterodimeric FSHR and LHR ligands
Summary and future prospects 197
Samenvatting 205
List of Publications 209
Curriculum Vitae 211
Nawoord 213
6
List of Abbreviations
5-HT2R serotonin receptor type 2 [] specific rotation
[(deg·mL)/(g·dm)]
Å angstrom(s) Abu aminobutyric acid AC adenylyl cyclase Ac acetyl
ADP adenosine diphosphate AIBN 2, 2’-azobis(isobutyronitrile) Ahx aminohexanoic acid Anhyd anhydrous
Ar aryl
Aq aqueous
ATP adenosine triphosphate Azp 4-azidoproline
B2R bradykinin receptor type 2
2AR 2-Adrenergic receptor Bn benzyl
Boc tert-Butyloxycarbonyl
BOP benzotriazol-1-yloxytri (dimethylamino)phosphonium hexafluorophosphate
br broad (NMR)
BRET bioluminescence resonance energy transfer
calcd calculated cAMP cyclic adenosine mono-
phosphate
CCR chemokine receptors CD circular dichroism CDCl3 deuterated chloroform CDI carbonyldiimidazole cDNA complementary
deoxyribonucleic acid CHO cells Chinese Hamster Ovary cells cm-1 wavenumber(s)
conc concentration CRE cAMP response element
CV column volume
δ chemical shift (NMR) d doublet (NMR)
d day(s) DCE dichloroethane DCM dichloromethane
dd doublet of doublets (NMR) ddd double doublet of doublets
(NMR) deg degrees
DiPEA diisopropylethylamine DHQ dihydroquinoline DMAP 4-(N,N-dimethylamino)-
pyridine
DMEM dulbecco’s modified Eagle’s medium
DMF N,N-dimethylformamide DMSO dimethylsulfoxide
EC effective concentration Eq molar equivalents ER endoplasmatic reticulum ESI electronspray ionization
Et ethyl
EtOAc ethyl acetate
(E)YFP (enhanced) yellow fluorescence protein
Fmoc 9-Fluorenylmethoxycarbonyl FRET fluorescence resonance
energy transfer
FSH(R) follicle stimulating hormone (receptor)
GABAB -aminobutyric acid type B GAP GTPase accelerating proteins GDP guanosine diphosphate GEF guanine nucleotide exchange
factor
GFP green fluorescent protein GnRH(R) gonadotropin-releasing
hormone (receptor)
GPCR G-protein coupled receptor GpHR glycoprotein hormone
receptor
GRK G-protein receptor kinases GTP guanosine triphospate
7 h hour
HA hemagglutinin
hCG human chorionic gonadotropin
HCTU 2-(6-chloro-1H-benzo triazole-1-yl)-1,1,3,3-
tetramethylaminium hexafluorophosphate HPLC high performance liquid
chromatography HRMS high resolution mass
spectrometry
Hyp hydroxyproline
IC inhibitory concentration i iso
IP3 inositol 1,4,5-triphosphate IR infra red
J coupling constant LC-MS liquid chromatography –
mass spectrometry
LH(R) luteinizing hormone (receptor)
LHA luteinizing hormone receptor agonist
LMW low molecular weight m multiplet (NMR) M molar (moles per liter) Me methyl
MeO methoxy MeOH methanol MHz megahertz min minute(s)
MS mass spectrometry M2R muscarinic receptor type 2 m/z mass-to-charge ratio NBS N-bromosuccinimide n.a. not active
n.d. not determined NMM N-methylmorpholine NMP N-methylpyrrolidone NMR nuclear magnetic resonance NOE(SY) nuclear Overhauser effect
(spectroscopy)
nu nucleophile obsd observed on overnight
PBS phosphate-buffered saline
PE petroleum ether PEG polyethylene glycol Pip 4-pipecolic acid PKA protein kinase A PLC phospholipase C ppm parts per million (NMR) Pro proline
q quartet (NMR) quant quantitative
Rf retention factor (TLC) Rluc renilla luciferase protein RP reversed phase rt room temperature s singulet (NMR)
SAR structure activity relationships
sat saturated
SPPS solid phase peptide synthesis t tertiary
t triplet (NMR) tert tertiary
molar ellipticity (deg·cm2·dmol-1)
THF tetrahydrofuran THQ tetrahydroquinoline TIS triisopropylsilane TLC thin layer chromatography TFA trifluoroacetic acid TM transmembrane
tR retention time (in chromatography)
Trz triazole
TSH(R) thyroid stimulating hormone (receptor)
v/v volume per unit volume VFTM venus flytrap module V2R vasopressin receptors type 2
