Download Dimeric ligands for GPCRs involved in human reproduction:

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 (CWNWO) 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
General introduction
9
Chapter 2
Synthesis and evaluation of homodimeric GnRHR ligands
37
Chapter 3
Synthesis and evaluation of homodimeric GnRHR antagonists having a
rigid propargylated benzene core
59
Chapter 4
GnRHR binding and antagonism of dimeric systems appear dependent
on the nature of the parent pharmacophore
83
Chapter 5
Discovery of selective LHR agonists by the bivalent ligand method
101
Chapter 6
Hydroxylated prolines as spacers for dimeric LHR agonists
119
Chapter 7
139
Oligoproline helix as a scaffold for potent, selective and
structurally defined dimeric ligands for the LHR
Chapter 8
Synthesis and pharmacological evaluation of dimeric FSHR antagonists
159
Chapter 9
Synthesis and pharmacological evaluation of heterodimeric
FSHR and LHR ligands
183
Summary and future prospects
197
Samenvatting
205
List of Publications
209
Curriculum Vitae
211
Nawoord
213
List of Abbreviations
5-HT2R
[]
Å
Abu
AC
Ac
ADP
AIBN
Ahx
Anhyd
Ar
Aq
ATP
Azp
B2R
2AR
Bn
Boc
BOP
br
BRET
calcd
cAMP
CCR
CD
CDCl3
CDI
cDNA
CHO cells
cm-1
conc
CRE
CV
δ
d
6
serotonin receptor type 2
specific rotation
[(deg·mL)/(g·dm)]
angstrom(s)
aminobutyric acid
adenylyl cyclase
acetyl
adenosine diphosphate
2, 2’-azobis(isobutyronitrile)
aminohexanoic acid
anhydrous
aryl
aqueous
adenosine triphosphate
4-azidoproline
bradykinin receptor type 2
2-Adrenergic receptor
benzyl
tert-Butyloxycarbonyl
benzotriazol-1-yloxytri
(dimethylamino)phosphonium
hexafluorophosphate
broad (NMR)
bioluminescence resonance
energy transfer
calculated
cyclic adenosine monophosphate
chemokine receptors
circular dichroism
deuterated chloroform
carbonyldiimidazole
complementary
deoxyribonucleic acid
Chinese Hamster Ovary cells
wavenumber(s)
concentration
cAMP response element
column volume
chemical shift (NMR)
doublet (NMR)
d
DCE
DCM
dd
ddd
deg
DiPEA
DHQ
DMAP
DMEM
DMF
DMSO
EC
Eq
ER
ESI
Et
EtOAc
(E)YFP
Fmoc
FRET
FSH(R)
GABAB
GAP
GDP
GEF
GFP
GnRH(R)
GPCR
GpHR
GRK
GTP
day(s)
dichloroethane
dichloromethane
doublet of doublets (NMR)
double doublet of doublets
(NMR)
degrees
diisopropylethylamine
dihydroquinoline
4-(N,N-dimethylamino)pyridine
dulbecco’s modified Eagle’s
medium
N,N-dimethylformamide
dimethylsulfoxide
effective concentration
molar equivalents
endoplasmatic reticulum
electronspray ionization
ethyl
ethyl acetate
(enhanced) yellow
fluorescence protein
9-Fluorenylmethoxycarbonyl
fluorescence resonance
energy transfer
follicle stimulating hormone
(receptor)
-aminobutyric acid type B
GTPase accelerating proteins
guanosine diphosphate
guanine nucleotide exchange
factor
green fluorescent protein
gonadotropin-releasing
hormone (receptor)
G-protein coupled receptor
glycoprotein hormone
receptor
G-protein receptor kinases
guanosine triphospate
h
HA
hCG
HCTU
HPLC
HRMS
Hyp
IC
i
IP3
IR
J
LC-MS
LH(R)
LHA
LMW
m
M
Me
MeO
MeOH
MHz
min
MS
M2R
m/z
NBS
n.a.
n.d.
NMM
NMP
NMR
NOE(SY)
nu
obsd
on
PBS
hour
hemagglutinin
human chorionic
gonadotropin
2-(6-chloro-1H-benzo
triazole-1-yl)-1,1,3,3tetramethylaminium
hexafluorophosphate
high performance liquid
chromatography
high resolution mass
spectrometry
hydroxyproline
inhibitory concentration
iso
inositol 1,4,5-triphosphate
infra red
coupling constant
liquid chromatography –
mass spectrometry
luteinizing hormone
(receptor)
luteinizing hormone receptor
agonist
low molecular weight
multiplet (NMR)
molar (moles per liter)
methyl
methoxy
methanol
megahertz
minute(s)
mass spectrometry
muscarinic receptor type 2
mass-to-charge ratio
N-bromosuccinimide
not active
not determined
N-methylmorpholine
N-methylpyrrolidone
nuclear magnetic resonance
nuclear Overhauser effect
(spectroscopy)
nucleophile
observed
overnight
phosphate-buffered saline
PE
PEG
Pip
PKA
PLC
ppm
Pro
q
quant
Rf
Rluc
RP
rt
s
SAR
sat
SPPS
t
t
tert
THF
THQ
TIS
TLC
TFA
TM
tR
Trz
TSH(R)
v/v
VFTM
V2R
petroleum ether
polyethylene glycol
4-pipecolic acid
protein kinase A
phospholipase C
parts per million (NMR)
proline
quartet (NMR)
quantitative
retention factor (TLC)
renilla luciferase protein
reversed phase
room temperature
singulet (NMR)
structure activity
relationships
saturated
solid phase peptide synthesis
tertiary
triplet (NMR)
tertiary
molar ellipticity
(deg·cm2·dmol-1)
tetrahydrofuran
tetrahydroquinoline
triisopropylsilane
thin layer chromatography
trifluoroacetic acid
transmembrane
retention time (in
chromatography)
triazole
thyroid stimulating hormone
(receptor)
volume per unit volume
venus flytrap module
vasopressin receptors type 2
7