Discovery of SAR184841, a potent and long-lasting inhibitor of 11β-hydroxysteroid dehydrogenase type 1, active in a physiopathological animal model of T2D

Article abstract of DOI:10.1016/j.bmcl.2013.02.018

Starting from 11β-HSD1 inhibitors that were active ex vivo but with Cyp 3A4 liability, we obtained a new series of adamantane ureas displaying potent inhibition of both human and rodent 11β-HSD1 enzymes, devoid of Cyp 3A4 interactions, and rationally designed to provide long-lasting inhibition in target tissues. Final optimizations lead to SAR184841 with good oral pharmacokinetic properties showing in vivo activity and improvement of metabolic parameters in a physiopathological model of type 2 diabetes.

Full text of DOI:10.1016/j.bmcl.2013.02.018

Bioorganic & Medicinal Chemistry Letters 23 (2013) 2414–2421
Contents lists available at SciVerse ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Discovery of SAR184841, a potent and long-lasting inhibitor
of 11b-hydroxysteroid dehydrogenase type 1, active in a
physiopathological animal model of T2D
a
a
a
a
a
Olivier Venier a, , Cécile Pascal , Alain Braun , Claudie Namane , Patrick Mougenot , Olivier Crespin ,
⇑
François Pacquet a, Cécile Mougenot a, Catherine Monseau a, Bénédicte Onofri a, Rommel Dadji-Faïhun a,
Céline Leger a, Majdi Ben-Hassine a, Thao Van-Pham a, Jean-Luc Ragot a, Christophe Philippo a,
Géraldine Farjot a, Lionel Noah a, Karima Maniani a, Asma Boutarfa a, Eric Nicolaï a, , Etienne Guillot ,
a
⇑
Marie-Pierre Pruniaux a, Stefan Güssregen b, Christian Engel b, Anne-Laure Coutant c, Beatriz de Miguel d,
Antonio Castro d
a Sanofi R&D, 1 Avenue Pierre Brossolette, 91385 Chilly-Mazarin, France
b Sanofi Deutschland GmbH, Industriepark Hoechst, 65926 Frankfurt am Main, Germany
c Sanofi R&D, 371 Rue du Professeur Joseph Blayac, 34184 Montpellier, France
d Sanofi R&D, Avenida de Leganés 62, 28925 Alcorcón, Madrid, Spain
a r t i c l e i n f o
a b s t r a c t
Article history:
Starting from 11b-HSD1 inhibitors that were active ex vivo but with Cyp 3A4 liability, we obtained a new
series of adamantane ureas displaying potent inhibition of both human and rodent 11b-HSD1 enzymes,
devoid of Cyp 3A4 interactions, and rationally designed to provide long-lasting inhibition in target tis-
sues. Final optimizations lead to SAR184841 with good oral pharmacokinetic properties showing
in vivo activity and improvement of metabolic parameters in a physiopathological model of type 2
diabetes.
Received 14 November 2012
Revised 31 January 2013
Accepted 3 February 2013
Available online 22 February 2013
Keywords:
Ó 2013 Elsevier Ltd. All rights reserved.
11b-Hydroxysteroid-dehydrogenase type I
11b-HSD1 inhibitors
Metabolic syndrome
Adamantane urea
Pharmacodynamic ex vivo assay
DIO model
11b-Hydroxy steroid dehydrogenase type 1 (11b-HSD1) con-
verts the biologically inactive glucocorticoids, cortisone (in man)
and 11-dehydrocorticosterone (in rodents), into their biologically
active metabolites, respectively, cortisol and corticosterone, which
are known to act as functional antagonists of insulin in several tar-
get organs and tissues such as the liver, muscle, and adipose tissue
(Fig. 1).1 There is evidence for an excess of cortisol in tissues being
a primary driver of insulin resistance and a critical point for disease
intervention.2 Liver- or adipose tissue-specific overexpression of
11b-HSD1 in transgenic mice produces a phenotype closely resem-
bling human type 2 diabetes mellitus.3 Reduction of intracellular
corticosterone levels in rodents as a result of pharmacological inhi-
bition of 11b-HSD1 reverses manifestations of altered metabolic
parameters including ectopic fat storage, diabetes, dyslipidemia
and atherosclerosis.4 Both 11b-HSD1 expression and activity are
up-regulated in the adipose tissue of obese, insulin-resistant or
diabetic humans.5 Inhibition of 11b-HSD1 therefore offers poten-
tial as a novel therapy to lower intracellular cortisol concentrations
and thereby enhance insulin sensitivity and hepatic lipid catabo-
lism in type 2 diabetes, obesity, and hyperlipidemia. Two isoforms
of 11b-HSD produced from distinct genes are known: 11b-HSD1 is
primarily expressed in the liver and adipose tissue, the other iso-
form, 11b-HSD2, is located mainly in the kidney.6 11b-HSD2 con-
verts active glucocorticoids (such as cortisol and corticosterone)
into inactive cortisone or 11-dehydrocorticosterone. Any potential
drug aimed at inhibiting 11b-HSD1 should therefore be selective
for this isoform since inhibition of 11b-HSD2 may result in side ef-
fects such as sodium retention, hypokaliemia, and hypertension.7
In the last decade, extensive research by the pharmaceutical
industry and academic groups allowed the identification of several
classes of 11b-HSD1 inhibitors, which have been comprehensively
⇑
Corresponding authors.
E-mail address: olivier.venier@sanofi-aventis.com (O. Venier).
0960-894X/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.bmcl.2013.02.018

O. Venier et al. / Bioorg. Med. Chem. Lett. 23 (2013) 2414–2421
2415
NC
Cl
N
H
H
N
N
NH2
N
N
N
S
S
O
O
O
O
S
O
PF-915275
BVT.2733
O
HO2C
S
N
H
Figure 1. Interconversion of cortisone and cortisol by 11b-HSD1 and 11b-HSD2
N
N
enzymes.
N
S
N
H
O
AMG221
AZD4017
Figure 2. Representative 11b-HSD1 inhibitors.
reviewed.8 These included sulfonamides such as BVT.2733 or PF-
915275,9,10 thiazolones such as AMG221,11 and amide derivatives
such as AZD4017 (Fig. 2).12
Several compounds have reached clinical development phases,
the most advanced being INCB-13739 which completed phase 2
as add-on therapy on top of metformin, and showed positive ef-
fects on end point criteria at the dosage of 200 mg in a 12 week
study (HbA1C (À0.6%), fasting plasma glucose (À24 mg/dl) and
HOMA-IR (À24%)).13 Other companies, for example, Vitae pharma-
ceuticals, BMS, Roche, Pfizer and Lilly are also currently undergoing
early clinical development with 11b-HSD1 inhibitors. However,
some compounds have been discontinued: PF-915275 for formula-
tion issues,14 BVT-3498 and BVT-83370 for unknown reasons,15
MK-0916 and MK-0736 for insufficient activity in a 12 weeks clin-
ical trial.16 Potential explanations for these failures were proposed:
(i) an unsuitable tissue distribution with insufficient activity in fat
relative to liver and (ii) a short a duration of action in target tissues.
Furthermore, several classes of 11b-HSD1 inhibitors displayed lim-
ited activity on rodent enzymes,8,9,12 making it difficult to assess
them in appropriate animal models for optimal drug candidate
selection. In metabolic disease pathogenesis, 11b-HSD1 activity is
upregulated in adipose tissue and liver, thus inhibition in both tis-
sues is considered to be a prerequisite for producing significant
therapeutic benefits.8b Therefore, in order to identify new drug
candidates with an optimal PK/PD profile we focused our efforts
on compounds displaying inhibition of both human and rodent en-
zymes, thus allowing early ex vivo testing in rodents. Furthermore,
we optimized the SAR to obtain compounds with high levels of
inhibition and long durations of action in both liver and fat tissues,
to ensure sufficient reduction of local cortisol levels, with the
objective of a once daily oral administration.
particular, we focused our efforts on the adamantane motif. This
group has often been reported in 11b-HSD1 inhibitors as a good
pharmacophore to address the lipophilic cofactor pocket of the en-
zyme and appeared to be a good alternative to increase the dura-
tion of action by increasing the overall lipophilicity.18 Moreover
the substitution of an adamantyl moiety with a polar group (hy-
droxyl, amide, carboxylic acid, etc.) can provide additional interac-
tion points with the cofactor and enable modulation of metabolic
stability.19 To assess the relevance of this strategy in the quinoxa-
line urea series and determine which kind of polar group is fa-
voured, we prepared a first set of compounds (2–5) as described
in Scheme 1.
As shown in Table 1, replacement of pyrrolidine pyrazole by
trans 4-amino adamantan-1-ol is well tolerated leading to potent
activities on both human and mouse enzymes and lower potency
on the rat enzyme.20 Moreover, compound 2 does not inhibit Cyp
3A4, indicating that pyrrolidine pyrazole was the main contribu-
tor to the Cyp interaction in compound 1. ADME data for com-
pound 2 are also favourable, with moderate metabolic lability
and good estimated intestinal permeability. Moving from trans
to cis 4-amino adamantan-1-ol in compound 3 leads to a dra-
matic loss of potency. Replacement of the hydroxyl group by a
primary amide gave rise to compound 4 with equipotent inhibi-
tion in human and mouse enzymes and a similar ADME profile.
On the other hand, transformation of the alcohol function into a
primary carbamate, while also well tolerated, lead to a decrease
in metabolic stability.
The Cyp 3A4 issue solved, we evaluated compound 2 in the
ex vivo assay which is pivotal in our search for long-lasting 11b-
HSD1 inhibitors.23 At 10 mg/kg per os, compound 2 displayed only
moderate inhibition after 2 h in the liver (inh. = 40%) and subcuta-
neous fat (inh. = 30%).
NH
N
O
N
N
N
MeO
N
N
1
Other modulations were evaluated to improve the ex vivo activ-
ity, and the replacement of 4-methoxyphenyl group by a methyl
sulphonylpiperazine moiety led to a family of compounds showing
increased inhibition levels and duration, up to 16 h post dosing
(see Table 2). The synthetic routes for preparing compounds 6–9
are shown in Schemes 2–4.
Ex-vivo inh. at 30mpk 2h post dosing
Liver 90%
Fat 70%
h11βHSD1 IC50= 43nM
m11βHSD1 IC50= 49nM
r11βHSD1 IC50= 600nM
Cyp 3A4 Inh. IC50< 300nM
h11βHSD2 IC50>10.000nM
The direct replacement of the 4-methoxyphenyl in compound 2
by 4-methylsulfonylpiperazine gave derivative 6 with a good bal-
ance in activity between human and rodent enzymes. Its ADME
profile was favourable for in vivo testing with acceptable metabolic
stability and good estimated intestinal permeability when in
presence of the P-gp inhibitor elacridar. The fact of being a P-gp
substrate was not considered detrimental for in vivo activity,25
since compound 6 showed some activity in the liver after 16 h in
the ex vivo assay. The pyridine derivative 7 was as active as its
pyrimidine analog in the enzymatic assay, but displayed a better
activity in the liver ex vivo assay and interestingly also a significant
We have previously described a new class of human and mouse
11b-HSD1 inhibitors containing the quinoxaline urea core. In our
prior report17 compound 1 was shown to be a potent and selective
inhibitor of 11b-HSD1, which inhibited 11b-HSD1 activity ex vivo
in liver and adipose tissue. However, further profiling in drug
metabolism studies showed that compound 1 was a potent inhib-
itor of Cyp 3A4 metabolic activity, exhibiting an IC50 below
300 nM. Our strategy to reduce the interaction with Cyp 3A4 con-
sisted in replacing the pyrrolidine pyrazole moiety with different
groups reported recognize the 11b-HSD1 enzymatic cleft. In

2416
O. Venier et al. / Bioorg. Med. Chem. Lett. 23 (2013) 2414–2421
O
N
N
a
MeO
N
N
MeO
N
H
N
N
H
R
N
N
3
2 R= OH
3 R= OH
b
4 R= CONH2
5 R=
OCONH2
Scheme 1. Synthesis of inhibitors (2–5). Reagents and conditions: (a) trisphosgene,21 DIPEA, CH2Cl2 then 4-amino adamantan-1-ol (2, 19% & 3, 31% or trans 4-amino
adamantan-1-carboxamide (4, 59%); (b) (1) Trichloroacetyl isocyanate, CH2Cl2, (2) K2CO3, MeOH, 50 °C, (5, 55%).22
Table 1
Replacement of the pyrrolidine pyrazole moiety: compounds 2–5
O
R
N
N
MeO
N
N
Compd
R
11b-HSD1 IC50 (nM)
h/m/r
% Metabolic lability
h/m/r
Caco2 permeability
CYP 3A4 inh.
(10À7 cm sÀ1
)
IC50
(lM)
N
N
N
N
OH
2
3
4
5
5/15/110
1400/ND/ND
18/36/70
9/5/100
41/21/33
ND
248
>10
H
OH
ND
182
ND
ND
H
H
H
CONH2
49/27/32
89/96/70
>10
>10
CONH2
O
Table 2
Modulation of aryl/heterocycle group: compounds 6–9
O
Cy2
Cy1
N
N
N
R
H
Compd Cy1
Cy2
R
11b-HSD1 IC50
(nM)
% Metabolic
lability
Caco2 permeability
CYP 3A4
inh.
Mice ex vivo (30 mg/kg)
%inh.
(10À7 cm sÀ1
)
h/m/r
h/m/r
IC50
(l
M)
16 h post dosing
O
N
Liver 30%
Sc Fat 0%
S
N
N
N
N
N
N
N
N
6
7
8
9
OH
OH
OH
14/1/28
22/2/16
50/1/114
9/18/24
12/18/19
2/9/17
7/109a
79
>10
N
N
O
O
Liver 55%
Sc Fat 40%
S
>30
>10
>10
O
O
Liver 80%
Sc Fat 55%
S
194
30
O
O
Liver 55%
Sc Fat 30%
S
CONH2 6/5/24
3/22/26
O
ND: not determined.
a
Plus elacridar.
inhibition of 11b-HSD1 activity in fat tissues for the first time in
the series. Replacement of the pyridine by a phenyl ring gave com-
pound 8 without losing the inhibitory activity on the enzyme in
human and mouse assays, but decreased the inhibitory effect on
the rat enzyme. However the replacement of the hydroxyl moiety
by a carboxamide group, for example, compound 9 enabled a

O. Venier et al. / Bioorg. Med. Chem. Lett. 23 (2013) 2414–2421
2417
H
N
O
N
O
O
O
N
O
N
R
N
N
N
(d)
(b)
N
N
N
(a)
OH
N
N
H
N
N
N
O
10
N
N
Br
11
N
S
N
S
O
O
O
R=Boc
R=H
12
13
(c)
6
Scheme 2. Synthesis of inhibitor 6. Reagents and conditions: (a) BuLi, THF, 2-chloro-5-bromo-pyrimidine 14%; (b) 15, NaOtBu, Pd2dba3, S-Phos, toluene, 100 °C, 70%;24 (c) HCl
4 M dioxane, 84%; (d) trisphosgene, TEA, DMF then trans 4-aminoadamantan-1-ol, 50%.
H
O
N
Cl
R
N
Cl
N
H
N
N
N
(a)
N
N
O
(b)
(d)
OH
+
N
S
N
O
O
Br
N
O
N
14
15
N
S
N
O
N
16
N
S
N
S
O
O
O
7
R=Boc 17
18
(c)
R=H
Scheme 3. Synthesis of inhibitor 7. Reagents and conditions: (a) NaOtBu, Pd2dba3, S-Phos, toluene, 100 °C, 76%; (b) Boc-quinoxaline, NaOtBu, Pd2dba3, S-Phos, toluene, 100 °C,
77%; (c) HCl 4 M dioxane, 80%; (d) trisphosgene, TEA, CH2Cl2 then trans 4-aminoadamantan-1-ol, 44%.
O
S
O
S
a
b
HN
N R
N
N
N
Br
N
N
N
Br
O
O
21 R=Boc
22 R=H
20
19
c
d
O
S
O
N
N
N
N
8 R=OH
9 R=CONH2
N
H
R
O
Scheme 4. Synthesis of inhibitors 8 & 9. Reagents and conditions: (a) MeSO2Cl, TEA, CH2Cl2, 94%; (b) Boc-quinoxaline, NaOtBu, Pd(OAc)2, P(tBu)3, o-xylene, 150 °C, 70%; (c)
HCl 4 M dioxane, 100%; (d) trisphosgene, TEA, DMF then trans 4-aminoadamantan-1-ol (8, 71%) or phosgene, TEA, DMF trans 4-aminoadamantan-1-carboxamide (9, 74%).
return to good inhibition of rat 11b-HSD1. Both compounds 8 and 9
displayed good ADME profiles and long-lasting activity in the
ex vivo assay with a level of inhibition similar to pyridine 7. Finally,
no Cyp 3A4 inhibition was observed within this set of molecules.
Compound 7 was considered as a good lead for further optimi-
zation due to its overall favourable profile with well balanced
enzymatic activity on human and rodent enzymes, acceptable
ADME parameters and significant ex vivo activity in target tissues
16 h after treatment. Moreover, the pyridine ring could give oppor-
tunities to improve the intrinsic solubility or to form solubilising
salts.26 In consequence, the final modulations shown in Table 3
were made on the N-pyridylquinoxaline scaffold.
of the physicochemical properties, in order to obtain a well bal-
anced inhibition profile in liver and fat tissues. All compounds
were assayed ex vivo for enzymatic inhibition in the liver and fat
tissues 16 h after single oral administration at lower dose, for
example, 3 mg/kg, in order to identify candidates for in vivo eval-
uation in a physiopathological model.
Replacement of the methylsulfonyl group by a t-butyl group led
to t-butylpiperazine 23, which displayed similar in vitro enzymatic
inhibition profile, ADME parameters and ex vivo activity to com-
pound 7. This result reflects the fact that the piperazine substitu-
tion is very tolerant in terms of chemical structure modulation
and supports the hypothesis of the absence of critical interactions
between this part of the molecule and the 11b-HSD1 protein (see
below). The acetamide 24 was also found to be potent in enzymatic
assays, but despite an acceptable ADME profile, did not show any
The synthesis of compounds 23–28 is depicted in Schemes 5–7.
In the final optimization step, we focused the chemical modula-
tions on the two far ends of the scaffold. This allowed a fine tuning

2418
O. Venier et al. / Bioorg. Med. Chem. Lett. 23 (2013) 2414–2421
Table 3
Modulation of piperazine & adamantyl substitution: compounds 23–28
O
N
N
R1
N
N
N
R2
N
H
Compd
R1
R2
11b-HSD1 IC50 (nM)
h/m/r
% Metabolic lability
h/m/r
Caco2 permeability (10À7 cm sÀ1
)
CYP 3A4 inh.
Mice ex vivo (30 mg/kg) %inh.
16 h post dosing
IC50
(l
M)
O
S
Liver 47%
Sc Fat 20%
7
OH
19/9/25
18/19/12
79
>30
O
Liver 55%
Sc Fat 20%
23
24
OH
OH
9/8/18
14/6/20
5/14/18
34
ND
O
Liver 0%
Sc Fat 0%
11/11/65
10/141a
>30
O
S
Liver 65%
Sc Fat 40%
25
26
27
OH
9/21/21
4/6/7
9/22/23
2/13/15
1/18/31
98
>10
>30
>30
O
Liver 72%
Sc Fat 60%
CONH2
CONH2
3/97a
4/144a
O
S
Liver 75%
Sc Fat 50%
4/8/9
O
ND: not determined.
a
Plus elacridar.
H
O
N
R
boc
N
F
N
N
N
R
N
(a)
(d)
(b)
N
N
N
Br
N
N
28
N
N
N
N
Br
29
23 R=OH
27 R=CONH2
30 R=Boc
31 R=H
(c)
Scheme 5. Synthesis of inhibitors 23 & 27. Reagents and conditions: (a) BocQuinoxaline, tBuOK, NMP, 90%; (b) NaOtBu, Pd2dba3, S-Phos, toluene, 110 °C, 74%; (c) HCl 4 M
dioxane, 100%; (d) phosgene, TEA, CH2Cl2 then trans 4-aminoadamantan-1-ol, (23, 52%) or phosgene, TEA, CH2Cl2 then trans 4-aminoadamantan-1-carboxamide (27, 52%).
H
Boc
N
R
N
Boc
N
O
N
N
N
N
(c)
(e)
(a)
OH
N
N
N
N
N
N
N
N
N
Br
N
N
29
N
R
O
O
32 R=Cbz
33 R=H
34 R=Boc
24
(b)
(d)
35 R=H
Scheme 6. Synthesis of inhibitors 24. Reagents and conditions: (a) CBz-piperazine, NaOtBu, Pd2dba3, S-Phos, toluene, 100 °C, 74%; (b) H2, Pd/C, MeOH, 94%; (c) acetyl chloride,
TEA, CH2Cl2, 100%; (d) HCl 4 M dioxane, 100%; (e) phosgene, TEA, CH2Cl2 then trans 4-aminoadamantan-1-ol, 53%.

O. Venier et al. / Bioorg. Med. Chem. Lett. 23 (2013) 2414–2421
2419
Table 4
Profile of SAR184841
H
N
O
Boc
N
R
N
N
N
h
h-Erg
Cyp 3A4
inhibition induction (Cerep
profile)
Cyp
Selectivity
In vitro
R
(c)
(a)
11bHSD2 IC50
IC50 (nM) (nM)
clearance in h.
Hep
N
N
(mL hÀ1 10À6hep
N
N
O
N
4000
>10000 None
None
Clean
0.035
N
N
N
N
H
N
S
N
S
Table 5
O
O
O
Pharmacokinetic profile for SAR184841 in rat (3 mg/kg iv & 10 mg/kg po)
33
Cmax
(ng/mL)
in
plasma
(po)
AUC0-inf
(ng h/mL)
in plasma
(po)
Tissue/plasma
AUC0-inf ratio
liver/adipose/
brain (po)
Plasma
clearance
(L/h/kg)
Vdss
(L/
kg)
t1/2 in
plasma
(po)
F%
36 R=Boc
37 R=H
25 R=OH
(b)
27 R=CONH2
Scheme 7. Synthesis of inhibitors 25 & 27. Reagents and conditions: (a) cyclo-
propylsulfonyl chloride, TEA, CH2Cl2, 100%; (b) HCl 4 M dioxane, 97%; (c) triphos-
gene, TEA, CH2Cl2 then trans 4-aminoadamantan-1-ol, (25, 50%), or trisphosgene,
TEA, CH2Cl2/DMF then trans 4-aminoadamantan-1-carboxamide (27, 56%).
556
580
224/2.2/0.14
5.2
14
4.4
25
35000
30000
25000
20000
15000
10000
5000
0
inhibition of 11b-HSD1 in the ex vivo assay after 16 h post-dosing.
Changing the methyl sulphonamide into cyclopropylsulfonamide
led to compound 25 which displayed a nanomolar activity in the
in vitro assays, but also a good level of activity ex vivo.
Control (lean), vehicle p.o.
DIO (obese), vehicle p.o.
SAR184841 DIO, 1 mg/kg p.o.
SAR184841 DIO, 3 mg/kg p.o.
SAR184841 DIO, 10 mg/kg p.o.
It is worth noting that the two most potent inhibitors in the
ex vivo assay, compounds 26 and 27, bear a primary amide group
instead of the hydroxyl group at position 1 of the adamantane ring.
Introduction of this carboxamide motif generated single digit
nanomolar inhibitors in the enzymatic assays with a very well bal-
anced activity on human and rodent enzymes. These two com-
pounds showed favourable overall ADME profiles and despite
being P-gp substrates (with low permeability when tested alone
but high permeability in the presence of elacridar) both inhibited
11b-HSD1 activity by more than 50% in target tissues 16 h after
3 mg/kg oral administration making them valid candidates for fur-
ther in vivo evaluation.27
# #
-23%
*
Finally compound 26 (SAR184841) was selected as the best po-
tential candidate for the following reasons: (i) a better ex vivo
activity in fat, (ii) a lower molecular weight, and (iii) the presence
of a basic group (pKa = 8.5 for the t-butylpiperazine) amenable to
salt formation to improve solubility (solubility >2 mg/mL in aque-
ous buffer up to pH 4.5).
Figure 4. Oral glucose tolerance test (OGTT) performed after the 3rd week of
treatment in DIO mice (18 h-fasted animals), treated with SAR184841 (1, 3 or
10 mg/kg/day). AUC: area under the glucose curve (0–60 min post 2 g/kg oral
glucose challenge); n = 9–10 animals per group; ⁄p <0.05, versus DIO obese group
(Dunnett test); ##p <0.01, versus control lean group (Student t test).
Figure 3. Crystal structure of human 11b-HSD1 in complex with compound 26.28

2420
O. Venier et al. / Bioorg. Med. Chem. Lett. 23 (2013) 2414–2421
Table 6
Effect of SAR184841 treatment on metabolic parameters in DIO mice model
Biochemical parameters
Plasma glucose
(mg/dL)
Plasma insulin
(ng/mL)
Plasma leptin
(ng/mL)
Plasma triglycerides
(mg/mL)
Plasma cholesterol
(mg/dL)
Liver triglycerides
(mg/g)
Control lean
vehicle
DIO obese
vehicle
SAR184841,
1 mg/kg
SAR184841,
3 mg/kg
226
7
0.6 0.1
1.0 0.4
27.1 1.3
36.2 2.1##
33.8 2.9
38.4 1.9
35.0 1.8
82.7 3.9
154.3 8.6##
127.7 5.8*
125.1 7.1*
123.6 6.4*
10.6 0.9
297 15##
3.6 0.2##
2.8 0.4
22.5 2.4##
14.9 1.4**
12.5 0.8**
12.5 1.2**
210.5 15.4##
167.4 15.0
142.8 26.7
129.1 19.4*
263
9
251 19
2.3 0.4*
1.9 0.2**
SAR184841,
10 mg/kg
224.6 10*
*
p <0.05, versus DIO obese group (Dunnett test).
p <0.01, versus DIO obese group (Dunnett test).
p <0.01, versus control lean group (Student test).
**
##
We obtained a crystal structure of SAR184841 in complex with
human 11b-HSD1 (PDB Code 4HX5). Figure 3 shows a view of
SAR184841 bound in the active site.
10 mg/kg/day, SAR184841 improved several key metabolic param-
eters monitored in the study (Table 6). Notably, dose-dependent
reductions of plasma glucose and plasma insulin were observed.
These effects were associated with an improvement of glucose tol-
erance measured after 3 weeks (Fig. 4, significant decrease of À23%
in OGTT AUC at 10 mg/kg). Plasma leptin, and plasma cholesterol
were also significantly decreased by SAR184841 at all doses. Liver
triglyceride levels, but not plasma triglycerides, were significantly
reduced at 10 mg/kg. Whatever the dose, SAR148841 had neither
an effect on food intake, nor on body weight of the DIO animals.
Plasma levels of SAR18484 at the end of the study (about 22 h after
the last administration) were quite low (0 0, 3.8 9.3, and
2.6 1.8 nM, respectively, for dose 1, 3 and 10 mg/kg), reflecting
the good clearance of the compound in the body, even after a
month-treatment. Finally, liver function biomarkers were not al-
tered by any of the treatment doses after 4-weeks treatment, and
HPA axis biomarkers (corticosterone) were not modified by
SAR184841 treatment (data not shown).
In conclusion, we found in our series that a substituted adaman-
tyl group is a good bioisoster for a pyrolidine pyrazole moiety, and
thus avoids Cyp 3A4 inhibition. Replacement of a pyrimidine ring
by a pyridine and introduction of a piperazine group led to in-
creased activity in the ex vivo assay. Finally, fine modulation of
piperazine and adamantyl substitution resulted in the discovery
of SAR184841. This compound is a potent human/rodent mixed
inhibitor of 11b-HSD1, with long lasting activity in target tissues.
It showed an improvement of key metabolic endpoints in a phys-
iopathological mouse model of obesity and diabetes after 4 weeks
of oral treatment. On the basis of its overall profile, SAR184841 was
selected as candidate for clinical development. Further details of
these investigations will be reported in due course.
The inhibitor binds with its central amide carbonyl group to the
hydroxyl side chains of the key residues Tyr183 and Ser170
responsible for substrate ketone reduction. The adamantyl amide
binds in the upper binding pocket formed by the protein stretches
Ile121-Leu126 and Thr222-Val227, the Tyr183 side chain and the
nicotine amide end of the co-factor NADP(H). The terminal amide
group forms hydrogen bonds to the oxygen atom at the pyrophos-
phate of the co-factor and a water-mediated hydrogen bond to the
main chain nitrogen of Thr124. The tetrahydroquinoxaline points
toward the solvent access channel. In the monomers with com-
pletely resolved ligand the attached pyridyl-piperazine points
stick-like into this channel. The pyridyl nitrogen forms a hydrogen
bond to the main chain nitrogen of Leu217. We did not observe
well defined contacts of the terminal t-butylpiperazine to the pro-
tein, because the C-terminus of the adjacent monomer forming
part of the solvent access channel is not well resolved in these
monomers. Residues of mouse 11b-HSD1 with differences in se-
quence to the human enzyme are not in direct contact with the
inhibitor or are located in regions that are typically highly flexible.
We postulate that the mixed human/rodent activity can be ex-
plained by minor protein adaptations accommodating the ligand.
Further profiling of SAR184841 revealed that the properties of
this compound made it suitable for preclinical development. As
seen in Table 4, SAR184841 exhibited about 1000-fold selectivity
for 11b-HSD1 versus human 11b-HSD2 and presented an IC50 over
10
l
M
for the h-Erg channel. The compound was inactive
M) in Cyp 3A4 inhibition assays either in a competitive
(IC50 >50
l
or a non competitive manner. No Cyp inductions were observed
and the selectivity profile versus a panel of 120 receptors and en-
zymes (including glucocorticoid and mineralocorticoid receptors)
was excellent.29 SAR184841 has a low in vitro metabolic clearance
in human hepatocytes and was also found negative in an in vitro
teratogenicity assay (Fetax) and mutagenicity assay (AmesII).
The pharmacokinetic profile of SAR184841 in rat is presented in
Table 5. The compound is characterized by extensive plasma clear-
ance, a very large volume of distribution, a long apparent terminal
half-life value and an acceptable oral bioavailability.
The potent and long-lasting effect of SAR184841 was demon-
strated in a pharmacodynamic study in mice over 24 h after single
oral administration. A 3 mg/kg dosage inhibited over 90% of 11b-
HSD1 activity in both liver and fat until 7 h post-dosing, and then
60% of enzyme activity in the liver and 40% of enzyme activity in
fat up to 24 h after administration (not shown). The in vivo efficacy
of SAR184841 was then investigated in a diet induced obese (DIO)
mice model.30 When administered once daily for 4 weeks at 1, 3 or
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SunFire R column C18 3.5
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intertwined dimer where the C-terminus of one monomer forms part of the
binding pocket and of a solvent access channel of the other monomer. The
tetramer is formed by two loosely packed dimers. The solvent access channel
connects the ligand binding site with the open solvent at the tetramer
interface. In each dimer the ligand is completely resolved in one of the two
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29. Receptor binding assays performed by Cerep (www.cerep.com).
30. C57Bl/6J mice, 20 week-old, fed a high-fat/high-fructose diet, were treated po
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during 4 weeks; n = 9–10 animals per group.