1H-Pyrazolo[3,4-g]hexahydro-isoquinolines as potent GR antagonists with reduced hERG inhibition and an improved pharmacokinetic profile

Article abstract of DOI:10.1016/j.bmcl.2015.10.097

We report the further optimization of our series 1H-pyrazolo[3,4-g]hexahydro-isoquinoline sulfonamides as GR antagonists. By incorporating a heteroaryl ketone group at the ring junction, we have obtained compounds with excellent functional GR antagonism. Optimization of the sulfonamide substituent has provided compounds with a very desirable overall profile, including minimal hERG activity, good bioavailability and in vivo efficacy.

Full text of DOI:10.1016/j.bmcl.2015.10.097

Bioorganic & Medicinal Chemistry Letters 25 (2015) 5720–5725
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
1H-Pyrazolo[3,4-g]hexahydro-isoquinolines as potent GR antagonists
with reduced hERG inhibition and an improved pharmacokinetic
profile
a
b
b
b
b
Hazel J. Hunt a, , Joseph K. Belanoff , Emily Golding , Benoit Gourdet , Timothy Phillips , Denise Swift ,
⇑
Jennifer Thomas b, John F. Unitt b, Iain Walters b
a Corcept Therapeutics, 149 Commonwealth Drive, Menlo Park, CA 94025, USA
b Sygnature Discovery, BioCity, Pennyfoot Street, Nottingham NG1 1GF, UK
a r t i c l e i n f o
a b s t r a c t
Article history:
We report the further optimization of our series 1H-pyrazolo[3,4-g]hexahydro-isoquinoline sulfonamides
as GR antagonists. By incorporating a heteroaryl ketone group at the ring junction, we have obtained
compounds with excellent functional GR antagonism. Optimization of the sulfonamide substituent has
provided compounds with a very desirable overall profile, including minimal hERG activity, good
bioavailability and in vivo efficacy.
Received 18 August 2015
Revised 29 October 2015
Accepted 30 October 2015
Available online 31 October 2015
Ó 2015 Elsevier Ltd. All rights reserved.
Keywords:
Glucocorticoid receptor antagonists
1H-Pyrazolo[3,4-g]hexahydro-isoquinolines
Insulin resistance
hERG inhibition
Glucocorticoids (GCs) regulate a plethora of biological effects,
including metabolism, inflammation, immunity, skeletal growth,
cardiovascular function, cognition, and physiological homeostasis.
Excessive glucocorticoid activity leads to a large number of adverse
effects, including glucose intolerance, diabetes, abnormal fat distri-
bution, osteoporosis, skin atrophy, hypertension, impaired wound
healing, depression, psychosis and impaired cognition. GCs exert
their effects via the ubiquitously expressed glucocorticoid receptor
(GR), a member of the nuclear hormone receptor family. GR nor-
mally resides in the cytoplasm as part of a multimeric complex
that includes chaperone proteins such as HSP90. When a ligand
binds, GR undergoes a conformational change, dissociates from
the chaperone proteins, and the GR-ligand complex translocates
to the nucleus. GR can act either as a monomer or homodimer,
and has direct or indirect effects on a wide spectrum of transcrip-
tional activities. Direct effects include binding to glucocorticoid
response elements (GREs) in the promoter or intragenic region of
GR responsive genes, which results in activation (positive GRE) or
repression (negative GRE) of gene transcription. The interaction of
GR with positive GREs results in the recruitment of co-activators,
whilst binding to negative GREs results in the recruitment of
co-repressors. GR can also bind to other transcription factors, such
as NF-
B or AP-1, which themselves bind to DNA.1,2
j
Mifepristone (KorlymTM) (Fig. 1), a potent GR antagonist, has
been approved in the US for the treatment of patients suffering
from endogenous Cushing’s syndrome for whom surgery is not
an option, or for whom surgery has failed. In addition to exhibiting
potent GR antagonism, mifepristone is also a potent antagonist of
the progesterone receptor (PR) and its approved use as an abortifa-
cient drug depends on this activity. We are interested in the devel-
opment of selective GR antagonists, devoid of PR antagonism, for
the treatment of Cushing’s syndrome and other disorders in which
excess cortisol is implicated. We previously reported3 the identifi-
cation of a series of 1H-pyrazolo[3,4-g]hexahydro-isoquinolines 1,
shown in Figure 1. Various modifications of substituents Y and X
were investigated, and numerous potent and selective GR antago-
nists were identified. Representative compounds were tested for
oral bioavailability in rats and dogs, and most of them exhibited
low bioavailability in rats but improved bioavailability in dogs.
Unfortunately, the compounds with the highest bioavailability in
rats were not the most potent GR antagonists in our in vitro assays.
One of the most promising compounds included in our
previous publication was CORT108297, which was designated as
compound 47 in that report. As shown in Figure 1, CORT108297
possesses an ethyl ether substituent at the ring junction and a
trifluoromethylphenylsulfonamide. As reported previously, this
⇑
Corresponding author. Tel.: +44 (0)1903 742945.
E-mail address: hhunt@corcept.com (H.J. Hunt).
http://dx.doi.org/10.1016/j.bmcl.2015.10.097
0960-894X/Ó 2015 Elsevier Ltd. All rights reserved.

H. J. Hunt et al. / Bioorg. Med. Chem. Lett. 25 (2015) 5720–5725
5721
O
O
O
O
X
O
S
S
N
N
Y
N
N
N
OH
N
N
CF3
O
F
F
mifepristone
1
CORT108297
Figure 1. Glucocorticoid receptor antagonist structures.
compound exhibited excellent potency (Ki = 6.8 nM) in a reporter
gene assay. CORT108297 has demonstrated very encouraging
in vivo results in several animal models, including the olanzapine
inducedweight gain model in rats,4,5 thediet-inducedobesity model
in mice,6 a rapid onset diabetes model in rats,7 the 3xTg-AD mouse
model of Alzheimer’s disease,8 the forced swim test in rats,9 and
the Wobbler mouse model of amyotrophic lateral sclerosis (ALS).10
CORT108297 has also been shown to attenuate the deleterious
effects of electroconvulsive therapy in rats.11 A recent publication
by Zalachoras et al.12 describes differential targeting of brain stress
circuits by CORT108297, and provides evidence that the compound
acts as a GR agonist, rather than an antagonist, on some parameters.
We now wish to report further optimization of this series,
leading to the identification of several compounds that combine
excellent GR antagonism with an improved pharmacokinetic
profile in rats, a reduction in hERG inhibition, and efficacy in vivo.
The heteroaryl ketones were synthesized from the common
intermediate 2 as depicted in Scheme 1. Intermediate 2 was used
to synthesize CORT108297 and other analogs, and its synthesis
has been described previously.3 Compound 2 was reduced to pro-
vide aldehyde 3, and then addition of an appropriate heteroaryl
lithium reagent resulted in the formation of the alcohols 4, which
could be readily oxidized to the required ketones 5. Removal of the
BOC protecting group under acidic conditions furnished the amines
6, which were coupled with the appropriate sulfonyl chloride to
provide the desired compounds 7. Direct conversion of the ester
2 to the ketone 5 was accomplished for many analogs by using
excess aryl lithium reagent. For some analogs, it was more conve-
nient to install the sulfonamide before the ketone. In these cases,
compound 2 was deprotected under acidic conditions and the
resultant amine 8 was converted into the desired sulfonamide 9.
Addition of an appropriate heteroaryl lithium reagent then
provided the required compounds 7. This approach is depicted in
Scheme 2.
The in vitro assays used to characterize the biological activities
of the GR ligands included a fluorescence polarization (FP) GR
binding assay13 and a tyrosine amino transferase (TAT) induction
assay in human HepG2 cells.14 Further characterization included
TAT induction assays in rat H4IIEC3 cells, and in primary hepato-
cytes derived from humans, rats, dogs and cynomolgus monkeys.14
Dexamethasone induces TAT activity in various cells, including
HepG2 cells, H4IIEC3 cells and hepatocytes. Compounds were
tested for their ability to antagonize the effect of dexamethasone
(measure of GR antagonism), and were also tested in the absence
of dexamethasone for their ability to induce TAT activity (measure
of GR agonism). hERG inhibition was evaluated in a manual patch
clamp assay.
Our objectives in continuing the optimization of our series of GR
antagonists were two-fold: we wanted to identify compounds with
improved GR antagonism that were devoid of GR agonist effects,
and we wanted to reduce the propensity for hERG inhibition.
CORT108297 has an IC50 of 0.8 lM in our manual patch clamp
assay. In the first stage of our optimization we maintained the tri-
fluoromethylphenylsulfonamide present in CORT108297 and
focused on varying the substituent at the ring junction. Based on
assessment of our model of CORT108297 docked in the active site
of GR (Fig. 2), we believed that increasing the size of this sub-
stituent would lead to improved antagonism, and abolish agonism.
We believe that the substituent at the ring junction in our series of
1H-pyrazolo[3,4-g]hexahydro-isoquinolines performs the same
role as the dimethylaminophenyl substituent in mifepristone.
The substituted phenyl group in mifepristone has been reported
to cause a shift in the position of helix 1215 and thus prevent the
formation of the conformation of GR required for agonism.
O
O
O
Ar
O
O
Ar
O
tBu
tBu
H
N
O
N
O
N
N
N
f
d
N
N
N
N
2
5
6
a
F
F
c
e
F
Ar
O
O
O
H
O
O
Ar
OH O
tBu
tBu
S
N
Y
N
O
N
O
N
N
N
b
N
N
N
3
4
7
F
F
F
Scheme 1. Reagents and conditions: (a) DiBAL, CH2Cl2, À78 °C (32%); (b) ArBr, nBuLi, ether, À78° or ArH, nBuLi, ether, À78 °C (13–29%); (c) Dess–Martin periodinane, CH2Cl2,
i
rt (quant.); (d) HCl, dioxane, rt or TFA, CH2Cl2, rt (quant.); (e) YSO2Cl, Pr2EtN, CH2Cl2, rt (50–80%); (f) ArBr, nBuLi, ether, À78 °C (40–70%).

5722
H. J. Hunt et al. / Bioorg. Med. Chem. Lett. 25 (2015) 5720–5725
O
O
O
O
O
O
O
O
O
tBu
H
S
N
O
N
N
Y
N
N
a
b
N
N
N
N
9
2
8
c
F
F
F
Ar
O
O
O
S
N
Y
N
N
7
F
Scheme 2. Reagents and conditions: (a) HCl, dioxane, rt or TFA, CH2Cl2, rt (quant.); (b) YSO2Cl, iPr2EtN, CH2Cl2, rt (40–90%); (c) ArBr, nBuLi, ether, À78 °C, or ArH, nBuLi, ether,
À78 °C (40–80%).
Table 1
GR binding and activity in HepG2 TAT assay for selected compounds
Ar
O
O
O
S
N
N
N
CF3
F
Compound Ar
GR binding Ki
HepG2 TAT Ki
(nM)
(nM)
10
11
12
13
14
2-Pyridyl
2-Imidazolyl
3-Pyridyl
2-Thiazolyl
5-Methyl-2-
oxadiaxolyl
4-Oxazolyl
2-Furanyl
2-Thienyl
1-Methyl-2-triazolyl
4-Thiazolyl
Pyrazinyl
5-Thiazolyl
0.16
0.47
0.83
0.23
1.6
14
52
151
18
129
15
16
17
18
19
20
21
0.16
0.17
0.41
0.4
0.14
0.21
0.33
63
31
27
117
22
98
Figure 2. CORT108297 (blue) overlaid with mifepristone (pink) in the GR active
site. Helix 12 shown in red. The GR structure comes from PDB 3H52, chain c.
CORT108297 was placed onto mifepristone using an RMS fitting.
We chose to investigate the incorporation of a range of hetero-
aryl ketones, and the results are summarized in Table 1. Most of
the analogs prepared exhibited excellent affinity for GR in our FP
binding assay, but potency in the cell based TAT assay was more
variable. Relatively lipophilic heteroaryl groups such as pyridyl
(compound 10) and thiazolyl (compound 13) were superior to
more polar heteroaryl groups such as imidazolyl (11) and pyrazinyl
(20). The position of attachment of the heteroaryl group was also
important; 2-pyridyl (10) was better than 3-pyridyl (12), and
2-thiazolyl (13) or 4-thiazolyl (19) were better than 5-thiazolyl
(21). We selected the 2-pyridyl analog 10 for further profiling,
including testing in TAT assays in the rat cell line H4IIEC3
(Ki = 4.2 nM) and in primary hepatocytes from several species.
Compound 10 demonstrated full antagonism in human and
monkey hepatocytes, partial antagonism in rat hepatocytes, and
full agonism in dog hepatocytes. These results are summarized in
Table 2. We previously observed similar differences between
species for CORT10829716, so agonism in the dog appears to be a
feature of this series of compounds. As anticipated, based on
results with earlier compounds in this series, Compound 10 was
selective for GR over the related nuclear hormone receptors MR,
PR, AR and ER (data not shown).
260
Table 2
Cross species TAT assay results for compound 10
Species
Antagonist Ki (nM)
Max
Agonist EC50 (nM)
Emax (%)
Human
Rat
Dog
100
13
—
110
62
—
280
1,600
51
110
Monkey
765
100
of the pharmacokinetic profile of the 2-pyridyl analog 10 dispersed
this concern. We administered the compound orally, formulated in
10% DMSO in 0.5% aqueous methyl cellulose, at a dose of 5 mg/kg,
and intravenously formulated in 10% DMSO, 45% PEG400 and 45%
propylene glycol at a dose of 1 mg/kg. We were gratified to dis-
cover that this compound achieved good plasma levels following
oral dosing, and had acceptable bioavailability (31%). For our initial
in vivo study we used our previously described rat model of olan-
zapine-induced weight gain.4 As previously reported4, female rats
administered olanzapine (3 mg/kg, bid) for 21 days gained more
weight than rats given vehicle for the same period. When dosed
orally twice a day for 21 days at a dose of 20 mg/kg (formulated
We had some concern that the inclusion of the ketone carbonyl
group might lead to a poor pharmacokinetic profile, but evaluation

H. J. Hunt et al. / Bioorg. Med. Chem. Lett. 25 (2015) 5720–5725
5723
in 10% DMSO, 0.1% Tween 80, 89.9% hypromellose), compound 10
prevented the olanzapine-induced weight gain, as shown in
Figure 3.17 The body weight of the rats administered compound
10 and olanzapine differed significantly (p <0.05) from the body
weight of rats given olanzapine alone from day 4 onwards.
CORT108297 at the same dose in the same vehicle was included
as a positive control, and was also effective from day 4 onwards.
The role of GR activation in the weight gain associated with the
use of atypical antipsychotic drugs such as olanzapine has not been
fully delineated. We wanted to demonstrate efficacy in an in vivo
model that has been widely accepted as GR mediated. Repeated
administration of cortisone to rats for six days results in elevated
glucose and insulin levels, and provides a convenient short dura-
tion model in which to assess the efficacy of our GR antagonists.
Cortisone is converted into cortisol in vivo by 11b-HSD1, and
although cortisol is not the natural GR agonist in rats, it binds to
the rat GR and acts similarly to the natural ligand, corticosterone.
Mifepristone was included as a positive control, and the results
are shown in Figure 4.18 As anticipated, rats administered cortisone
(30 mg/kg sc) once a day for six days exhibited higher glucose and
insulin levels than control rats that did not receive cortisone.
Mifepristone given orally twice a day at a dose of 30 mg/kg pre-
vented the cortisone-induced elevation in glucose and insulin
levels, and compound 10 at the same dose attenuated the increase
in glucose, but did not have a significant effect on insulin levels.
Compound 10 (CORT113176) has also been tested by Beaudry
et al.7 in their rapid onset diabetes model in rats, and encouraging
results were obtained. Compound 10 improved fasting glycaemia
and acute insulin response and attenuated peripheral insulin
resistance. Vendruscolo et al.19 have recently reported that
CORT113176 reduces alcohol intake in rats made dependent on
alcohol.
Encouraged by these results, we next undertook optimization of
the phenyl sulfonamide. For our initial exploration, we maintained
the 2-pyridyl ketone substituent present in compound 10. One of
our objectives in this phase of the project was the reduction of
inhibition of hERG activity. In a manual patch clamp assay, an
IC50 of 5 lM was determined for compound 10. Conscious that
compounds such as 10 have a relatively high molecular weight,
we focused predominantly on the inclusion of substituents that
would not contribute excessive additional weight. We incorpo-
rated several small substituents, including fluoro, chloro, methyl,
methoxy and cyano and also investigated the effect of ortho, meta
or para substitution. Most of the compounds prepared possessed
excellent affinity in the FP binding assay, and good GR antagonism
in the dexamethasone induced TAT assay in human HepG2 cells.
Details of selected analogs are provided in Table 3. We anticipated
that reducing the lipophilicity of the compound, by suitable
manipulation of the substituent R, would provide a reduction in
hERG inhibition. We were surprised to discover that replacing
the para trifluoromethyl substituent present in compound 10 with
a para fluoro substituent resulted in increased hERG inhibition—
compound 22 had an IC50 of 0.98 lM in a manual patch clamp
assay. Another surprising discovery was the effect of the position
of substitution on hERG inhibition. Moving the trifluoromethyl
substituent from the para position to the meta position achieved
a substantial reduction of hERG inhibition with compound 23 hav-
ing an IC50 > 20
hERG assay precluded testing at a higher concentration. Compound
23 achieved only 29% inhibition at a concentration of 10 M and
11% inhibition at a concentration of 1 M. In order to determine
lM. Limited solubility in the medium used for the
l
l
whether this effect was general, we investigated other meta substi-
tuted compounds. Analogs incorporating meta methyl (compound
24), meta chloro (compound 25) or meta methoxy (compound 26)
Weight
Olanzapine
Body weight (g)
Vehicle
CORT108297
CORT113176
290
280
270
260
250
240
230
220
0
5
10
Day of study
15
20
Figure 3. Effects of compound 10 (CORT113176) in the olanzapine-induced weight gain model.
Figure 4. Effects of compounds 10 and 33 in the cortisone induced insulin elevation model. A = vehicle + vehicle; B = cortisone + vehicle; C = cortisone + mifepristone;
D = cortisone + compound 10; E = cortisone + compound 33. ***p <0.001 compared to group A; ###p <0.001 compared to group B, #p <0.05 compared to group B.

5724
H. J. Hunt et al. / Bioorg. Med. Chem. Lett. 25 (2015) 5720–5725
Table 3
on the PK profile, we synthesized several disubstituted compounds
Variation of the phenyl sulfonamide
(see Table 3). We were pleased to discover that by adding a meta
substituent to a para substituted compound we were able to miti-
gate hERG inhibition. For example, the para fluoro, meta trifluo-
romethyl analog 30 had significantly reduced hERG inhibition
compared with compound 22 which lacked the meta substituent.
Even the addition of a meta fluoro substituent was sufficient to
reduce hERG inhibition, as shown by compound 31. We were dis-
appointed to discover that compound 30 did not exhibit an
improved PK profile compared to compound 23. There was no
improvement in Cmax after oral dosing (112 ng/ml for compound
30 compared to 155 ng/ml for compound 23) and compound 30
actually suffered from increased clearance (103 ml/min/kg) after
iv administration. The PK profile of compound 31 was more
encouraging, with Cmax 398 ng/ml after an oral dose of 5 mg/kg
and clearance 44 ml/min/kg after an iv dose of 1 mg/kg. We discov-
ered that one of the best combinations was a meta fluoro sub-
stituent and a para trifluoromethyl group, compound 32. This
combination provided minimal hERG inhibition and a good PK pro-
file, with Cmax 322 ng/ml after oral dosing and clearance 28.7 ml/
min/kg after i.v. administration. The meta, para dichloro compound
33 also provided an excellent combination of minimal hERG inhi-
O
O
O
N
S
N
R
N
N
F
Compound
R
Substitution GR
HepG2
hERG
inhibition
binding Ki TAT Ki
(nM)
(nM)
10
22
23
CF3
F
CF3
p
p
0.16
0.13
0.1
14
16
14
IC50 = 5
IC50 = 0.98 lM
IC50 >20
11% @ 1
29% @ 10
17% @ 1
30% @ 10
14% @ 1
35% @ 10
IC50 >19
24% @ 1
36% @ 10
34% @ 1
59% @ 10
27% @ 1
42% @ 10
26% @ 1
45% @ 10
lM
m
lM
l
M
l
M
24
25
26
CH3
Cl
m
m
m
0.14
0.25
0.19
6.8
11
l
M
l
M
l
M
l
M
OCH3
11.4
lM
l
M
bition (IC50 > 10 lM) and good oral bioavailability in rats (Cmax
l
M
437 ng/ml after a 5 mg/kg dose). GR antagonism in the rat cell line
H4IIEC3 (Ki = 8.5 nM) and primary rat hepatocytes (Ki 150 nM, 74%
efficacy) was confirmed. Compound 33 was tested in the cortisone
induced insulin resistance model in rats (30 mg/kg orally, twice a
day). This compound achieved a statistically significant reduction
in both glucose and insulin levels, and the results are included in
Figure 3.
In summary, we have improved our original series of 1H-pyra-
zolo[3,4-g]hexahydro-isoquinoline GR antagonists by the incorpo-
ration of a heteroaryl ketone substituent, and the modification of
the sulfonamide portion. Structure activity relationship studies
have revealed that the inclusion of a meta substitution the phenyl
sulfonamide provides compounds with a substantial reduction in
hERG liability. The combination of meta and para substituents pro-
vides the optimum balance of hERG activity and oral bioavailabil-
ity. Compound 33 has demonstrated promising in vivo activity in a
rat model of insulin resistance. Further optimization of our series
of 1H-pyrazolo[3,4-g]hexahydro-isoquinolines and the identifica-
tion of our clinical candidate CORT125134 will be reported in
due course.
27
28
29
30
31
32
33
CH3
CN
p
0.22
0.14
0.21
0.73
0.22
0.29
0.64
4.5
15
l
M
l
M
m
o
l
M
l
M
CH3
18.5
9.7
7.5
11
l
M
l
M
CF3
F
F
F
CF3
F
m
p
m
p
16% @1
31% @10 lM
10% @1 lM
35% @10 lM
8% @1 lM
17% @10 lM
lM
p
m
m
p
Cl
Cl
14
IC50 ꢀ10
l
M
7% @ 1
lM
16% @ 10 lM
substitution also exhibited reduced hERG inhibition, with 30%, 35%
and 36% inhibition at 10 M, respectively. In comparison, com-
pound 27 with the methyl substituent in the para position showed
59% inhibition at 10 M and 34% inhibition at 1 M. The more
polar cyano substituent did not provide much benefit, with meta-
substituted compound 28 achieving 42% inhibition at 10 M. Ortho
l
l
l
l
Acknowledgements
substitution generally led to reduced GR antagonist potency, for
example, compare compound 29 with an ortho methyl substituent
to the corresponding meta and para substituted analogs 24 and 27.
There did not appear to be any advantage in terms of hERG inhibi-
The authors would like to thank Argenta Discovery (now part of
Charles River) for the original synthesis of a few of the compounds
described in the manuscript. The authors would also like to thank
ICBC for conduction of the manual patch clamp hERG assays, and
Saretius for carrying out the rat PK studies. In addition, the authors
would like to thank Dr Colin Sambrook Smith for providing
Figure 2.
tion, with compound 29 exhibiting 45% inhibition at 10 lM.
Encouraged by the reduced hERG inhibition exhibited by these
compounds, we evaluated several meta substituted analogs in PK
studies in rats. We were disappointed to discover that moving
the trifluoromethyl substituent from the para (compound 10) to
the meta (compound 23) position had a detrimental impact on
the PK profile. The Cmax achieved after administering a 5 mg/kg oral
dose was decreased from 375 ng/ml to 155 ng/ml, and clearance
after iv dosing was increased from 11 ml/min/kg to 40 ml/min/
kg. Similarly, the meta chloro analog 25 suffered from a low Cmax
(112 ng/ml) after oral dosing and a high clearance (70 ml/min/kg)
after iv dosing. Although we did not carry out any studies to deter-
mine the metabolic fate of these meta substituted compounds, we
reasoned that the lack of a substituent at the para position con-
tributed to the high clearance observed.
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resulting in a decrease in the measured polarisation value. The change in
polarisation value in the presence of test compounds is used to calculate the
binding affinity of the compound for GR.
14. TAT activity was measured as described by Ali, A.; Thompson, C. F.; Balkovec, J.
M.; Graham, D. W.; Hammond, M. L.; Quraishi, N.; Tata, J. R.; Einstein, M.; Ge,
L.; Harris, G.; Kelly, T. M.; Mazur, P.; Pandit, S.; Santoro, J.; Sitlani, A.; Wang, C.;
Williamson, J.; Miller, D. K.; Thompson, C. M.; Zaller, D. M.; Forrest, M. J.;
Carballo-Jane, E.; Luell, S. J. Med. Chem. 2004, 47, 2441.
11. Andrade, C.; Shaikh, S. A.; Naravan, L.; Blasey, C.; Belanoff, J. J. Neural Transm.
2012, 119, 337.
15. Schoch, G. A.; D’Arcy, B.; Stihle, M.; Burger, D.; Bar, D.; Benz, J.; Thoma, R.; Ruf,
A. J. Mol. Biol. 2010, 395, 568.
12. Zalachoras, I.; Houtman, R.; Atucha, E.; Devos, R.; Tijssen, A. M. I.; Hu, P.;
Lockey, P. M.; Datson, N. A.; Belanoff, J. K.; Lucassen, P. J.; Joels, M.; de Kloet, E.
R.; Roozendaal, B.; Hunt, H.; Meijer, O. C. P.N.A.S. 2013, 110, 7910.
13. The binding affinity of test compounds was determined using a FP binding
16. Hunt, H. J. Unpublished observations.
17. The rat olanzapine induced weight gain model was carried out by Saretius, UK.
18. The cortisone induced insulin resistance model was carried out by RenaSci, UK.
19. Vendruscolo, L. F.; Estey, D.; Goodell, V.; Macshane, L. G.; logrip, M. L.;
Schlosburg, J. E.; McGinn, M. A.; Zamora-Martinez, E. R.; Belanoff, J. K.; Hunt, H.
J.; Sanna, P. P.; George, O.; Koob, G. F.; Edwards, S.; Mason, B. J. J. Clin. Invest.
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assay using human recombinant GR (PanVera P2812) and
labelled glucocorticoid ligand (Fluorome GS Red) (PanVera P2894). The
presence of inhibitors prevents the formation of GS Red/GR complex
a fluorescent
a