Discovery and optimization of adamantyl carbamate inhibitors of 11β-HSD1

Article abstract of DOI:10.1016/j.bmcl.2010.08.142

Synthesis of 2-adamantyl carbamate derivatives of piperidines and pyrrolidines led to the discovery of 9a with an IC₅₀ of 15.2 nM against human 11β-HSD1 in adipocytes. Optimization for increased adipocyte potency, metabolic stability and select

Full text of DOI:10.1016/j.bmcl.2010.08.142

Bioorganic & Medicinal Chemistry Letters 20 (2010) 6725–6729
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Discovery and optimization of adamantyl carbamate inhibitors of 11b-HSD1
Colin M. Tice a, , Wei Zhao a, Paula M. Krosky a, Barbara A. Kruk a, Jennifer Berbaum a, Judith A. Johnson a,
⇑
Yuri Bukhtiyarov a, Reshma Panemangalore a, Boyd B. Scott a, Yi Zhao a, Joseph G. Bruno a, Lamont Howard a,
Jennifer Togias a, Yuan-Jie Ye a, Suresh B. Singh a, Brian M. McKeever a, Peter R. Lindblom a, Joan Guo a,
Rong Guo a, Herbert Nar b, Annette Schuler-Metz b, Richard E. Gregg a, Katerina Leftheris a,
Richard K. Harrison a, Gerard M. McGeehan a, Linghang Zhuang a, David A. Claremon a
a Vitae Pharmaceuticals, 502 W. Office Center Drive, Fort Washington, PA 19034, USA
b Boehringer Ingelheim Pharma GmbH & Co. KG, 88400 Biberach, Germany
a r t i c l e i n f o
a b s t r a c t
Article history:
Received 2 July 2010
Revised 28 August 2010
Accepted 31 August 2010
Available online 7 September 2010
Synthesis of 2-adamantyl carbamate derivatives of piperidines and pyrrolidines led to the discovery of 9a
with an IC50 of 15.2 nM against human 11b-HSD1 in adipocytes. Optimization for increased adipocyte
potency, metabolic stability and selectivity afforded 11k and 11l, both of which were >25% orally bio-
available in rat.
Ó 2010 Elsevier Ltd. All rights reserved.
Keywords:
11b-Hydroxysteroid dehydrogenase
Diabetes
11b-hydroxysteroid dehydrogenase (11b-HSD1) is a member of
the short chain dehydrogenase/reductase (SDR) superfamily.1,2
11b-HSD1 increases local tissue concentrations of the active gluco-
corticoid cortisol by NADPH-dependent reduction of the 11-keto
group of inactive cortisone to the (11S)-alcohol, cortisol. 11b-HSD1
is primarily expressed in liver and adipose tissue, and its elevated
expression in adipose tissue has been linked to obesity, insulin resis-
tance, metabolic syndrome, diabetes and cardiovascular disease in
humans.3–7 Moreover, the phenotype of transgenic mice over-
expressing 11b-HSD1 in adipose tissue includes visceral obesity,
insulin resistance and hypertension,8,9 while 11b-HSD1 knockout
mice are resistant to diet induced obesity and have increased insulin
sensitivity.10–12 Thus, a selective inhibitor of 11b-HSD1 may be use-
ful for the treatment of diabetes and has potential for positive effects
on multiple cardiovascular risk parameters.
Since 2002,13 several classes of selective, synthetic inhibitors of
11b-HSD1 have been reported (Fig. 1).14–16 Triazole 1 increased
insulin sensitivity and decreased fasting glucose, cholesterol, and
adipose tissue mass in mice.17 Oral dosing of thiazolone 2 in-
creased plasma adiponectin levels and decreased fasting glucose
Figure 1. Literature 11b-HSD1 inhibitors.
c
levels in KKA mice.18,19 Sulfonamide 3 reduced fed glucose and
fasted insulin in mice when incorporated into a high fat diet at
30 mg/kg/day.20 Racemic adamantyl amide 4 lowered plasma glu-
cose levels after 3 weeks of oral administration to ob/ob mice at
30 mg/kg b.i.d.21 The activity of these compounds in animal models
⇑
Corresponding author. Tel.: +1 215 461 2042; fax: +1 215 461 2006.
E-mail address: ctice@vitaerx.com (C.M. Tice).
Figure 2. Adamantyl urea and carbamate 11b-HSD1 inhibitors.
0960-894X/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2010.08.142

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C. M. Tice et al. / Bioorg. Med. Chem. Lett. 20 (2010) 6725–6729
Figure 3. Model of 6 in the 11b -HSD1 binding site. The surface of the dimeric enzyme shown with residues 213–233 excluded to provide a clear view of the active site. Tyr
177 is highlighted to show the hydrophobic interaction with the piperidine ring. The solvent accessible surface due to monomer A is colored according to the electrostatic
surface properties and that of monomer B is shown in magenta. The carbonyl of compound 6 interacts with the catalytic residues Tyr183 and Ser170.
further validates the potential for selective inhibition of 11b-HSD1
as a treatment for diabetes.
carbamates of readily available pyrrolidines and piperidines which
included 9a (Scheme 1).
We previously described the discovery of N-(2-adamantyl)ureas
such as 5 (Fig. 2).22 Our model of 5 bound to 11b-HSD1, based on the
ligand in PDB code 2IRW,23 indicated that the urea NH did not par-
ticipate in a hydrogen bond with the protein. Replacement of the
urea NH with O gave carbamate 6 resulting in a 3Â improvement
in enzyme potency but a 1.5Â decrease in cell potency. Our model
of 6 bound to 11b-HSD1 is depicted in Figure 3. Whereas the ada-
mantyl group filled the large, mainly hydrophobic pocket adjacent
to the cofactor (Pocket I), the piperidine ring of 6 only partially filled
Pocket II, suggesting that larger groups would be tolerated in this
region and might increase potency. The low MW of 6 (263 da) also
permitted additional substitution on the molecule, although the
high clog D7.0 (calculated log D at pH 7.0) and low PSA (polar surface
area) of the molecule dictated that polar substituents would be
required to improve the solubility and drug-likeness of the
compounds synthesized.24–26 The medicinal chemistry program
commenced with the synthesis of a small library of 2-adamantyl
The synthesis of analogs 9a–i is depicted in Scheme 1. 2-Ada-
mantyl alcohol 7 was converted to its chloroformate by treatment
with triphosgene in the presence of pyridine and reacted with
(3R)-(3-t-butoxycarbonylamino)pyrrolidine to afford carbamate
9a. Removal of the Boc group, followed by acylation of the resulting
amine, gave analogs 9b–g. Alternatively, the BocNH group of 9a
was alkylated with MeI or BnBr to afford 9h and 9i, respectively.
Similar chemistry was employed to prepare analogs of general
structure 10 in which the adamantane ring system was substituted
at the 5-position (Scheme 2). The E and Z adamantane isomers
were separated by preparative HPLC to afford 10a–c. In all cases,
the desired E isomer had a longer retention time on reverse phase
HPLC. The chemical shift of the proton on the oxygen-substituted
carbon of the adamantane in the E isomer was downfield by
ꢀ0.03 ppm compared to the Z isomer. The assignment of the E
geometry was subsequently confirmed when X-ray structures of
11k and 11l bound to 11b-HSD1 were obtained (data not shown).
Scheme 1. Synthesis of compounds 9 and 10. For complete substituent definitions see Tables 1 and 3. Reagents and conditions: (a) triphosgene, pyridine, CH2Cl2, 0–5 °C, 3 h;
(b) (3R)-(3-t-butoxycarbonylamino)pyrrolidine, i-Pr2NEt, CH2Cl2, 0 °C?rt, 16 h; (c) preparative HPLC (X – H); (d) 4 M HCl in dioxane, CH2Cl2, rt, 1 h or 1:4 TFA/CH2Cl2, rt, 1 h;
(e) R1COCl, i-Pr2NEt, CH2Cl2, 0 °C?rt; (f) MeI or BnBr, NaH, DMF; (g) LiBH4 to give 10d; (h) MeMgBr to give 10e; (i) LiOH, H2O, MeOH followed by NH3 in dioxane, EDC, HOBt, i-
Pr2NEt, CH2Cl2, 0 °C?rt, 16 h to give 10f; (j) (CF3CO)2O, pyridine, CH2Cl2, 0 °C to rt.

C. M. Tice et al. / Bioorg. Med. Chem. Lett. 20 (2010) 6725–6729
6727
Scheme 2. Methods for the synthesis of compounds of general structure 11. See Table 4 for definitions of Het. Reagents and conditions: (a) HCl, dioxane, CH2Cl2, rt, 1 h; (b)
Het-X, i-Pr2NEt, n-PrOH, , microwave; (c) 5-(methoxycarbonyl)-2-adamantyl chloroformate, i-Pr2NEt, CH2Cl2, 0 °C to rt; (d) LiOH, H2O, MeOH, rt, 16 h; (e) NH3 in dioxane,
D
EDC, HOBt, i-Pr2NEt, CH2Cl2, 0 °C?rt, 16 h; (f) preparative HPLC.
Reduction of ester 10c with LiBH4 led to primary alcohol 10d, while
treatment with excess MeMgBr gave tertiary alcohol 10e. Hydroly-
sis of the ester, followed by EDC coupling with ammonia, gave car-
boxamide 10f. Analog 10g, the (S) isomer of 10f, was prepared in
the same fashion employing (3S)-(3-t-butoxycarbonylamino)pyr-
rolidine. Dehydration of 10f afforded nitrile 10h.
Table 2
Calculated physical properties and biological data for selected compounds
PSAa (Å2)
Plasma
shiftb
RLM t1/2
(min)
a
c
Compound
MW (da)
clogD7.0
6
9a
10f
11k
11l
263
365
408
409
409
3.4
3.2
1.9
1.4
1.4
30
68
111
121
121
28.4
20.9
1.2
2.7
2.2
2
2
30
>60
46
Analogs of general structure 11 were prepared by two closely
related methods (Scheme 2). Removal of the Boc group from 10f,
followed by SNAr reaction of the primary amine with electron defi-
cient haloheterocycles gave 11 (Method A, Scheme 2). Alterna-
tively, Boc protected 3-aminopyrrolidine 12 was reacted with a
haloheterocycle under SNAr conditions and deprotected to afford
a 3-(heterocyclylamino)pyrrolidine 13 which was reacted with 1-
(methoxycarbonyl)-4-adamantyl chloroformate. Ester hydrolysis
and carboxamide preparation, followed by preparative HPLC to
separate the E and Z adamantane isomers, gave target compounds
11 (Method B, Scheme 2). Analogs with the (S) stereochemistry at
the 3-position of the pyrrolidine ring were prepared from 10g fol-
lowing Method A or from the (S) isomer of 12 following Method B.
The first phase of the medicinal chemistry program led to the
identification of 9a (Table 1) which had superior enzyme and cell
potency to the prototype compound 6. The presence of additional
polar atoms in 9a was considered to be an attractive feature
although the plasma shift was only marginally reduced compared
to 6, and metabolic stability remained poor (Table 2).
a
Calculated using Pipeline Pilot, Accelrys, San Diego, CA.
Ratio of enzyme IC50 measured in the presence and absence of 50% human
b
plasma.
c
RLM t1/2 is rat liver microsome half life. See Ref. 33.
was less active. A more substantial reduction in activity occurred
when the ether oxygen of the t-butoxy group in 9a was replaced
with a CH2 group to give amide 9g. These results suggested that
the t-butoxy group is optimal at R1 and that the ether oxygen plays
an important role. N-methylation (9h) was well tolerated, while N-
benzylation (9i) reduced potency.
Changes to the substituent on the pyrrolidine ring initially
failed to identify analogs with improved potency over 9a and rapid
metabolism of 9a was shown to occur predominantly on the ada-
mantyl ring system. Therefore, we turned our attention to modifi-
cations of the adamantane ring. Amides of 2-aminoadamantane
have previously been studied as inhibitors of 11b-HSD1.23,27–30 In
these molecules, polar substituents at the trans-5-position of the
adamantane ring were reported to be compatible with potency
while improving metabolic stability.29 Hydroxyadamantyl analog
10a retained comparable enzyme and cell potency to 9a, while
acetamido analog 10b was 6Â less active (Table 3). Ester 10c was
100Â less potent than 9a. Primary alcohol 10d regained consider-
able enzyme potency; however, tertiary alcohol 10e had little
activity. The carboxamide group in 10f improved cell potency to
7.5 nM. Compound 10g, the (S) isomer of 10f, was 2–3Â less potent
in cells than 10f. Nitrile 10h was also 3Â less potent in cells than
10f from which it was derived.
Compound 10f exhibited a reduction in clog D7.0 and an increase
in PSA compared to both 9a and the original carbamate 6 (Table 2).
Compound 10f was selected for further evaluation in vitro. The
compound suffered <2Â loss in enzyme potency in the presence
of 50% human plasma, had an improved RLM t1/2, and exhibited a
selective profile with IC50 values for the hydroxysteroid dehydro-
genases 11b-HSD2, 3b-HSD2 and 17b-HSD1 of >10,000 nM. Its
EC50 values against FXR and GR were >10,000 nM, and its IC50 val-
ues on CYP3A4, CYP2C9 and CYP2D6 were all >30,000 nM. Despite
these desirable features, 10f was deemed to have inadequate adipo-
cyte potency.
Replacement of the t-butoxy group in 9a with smaller alkoxy
groups (9b–d) reduced the enzyme potency. The isobutoxy
compound 9e was equiactive, while the benzyloxy compound 9f
Table 1
Carbamates 9
O
R1
N
O
N
R2
O
a
a
Compound
R1
R2
Enzyme IC50 (nM)
Adipocyte IC50 (nM)
6
5
53
9a
9b
9c
9d
9e
9f
9g
9h
9i
Ot-Bu
OEt
Oi-Pr
On-Pr
Oi-Bu
OBn
CH2t-Bu
Ot-Bu
Ot-Bu
H
H
H
H
H
H
H
Me
Bn
2.7
12.0
6.4
5.4
2.0
9.0
68
15.2
65.5
36.3
40.2
19.2
84.8
—
2.9
15.3
24.7
290
a
See Ref. 32.

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C. M. Tice et al. / Bioorg. Med. Chem. Lett. 20 (2010) 6725–6729
Table 3
Table 5
Substituted adamantyl compounds 10
Metabolic stability and CYP inhibition of heteroaryl compounds 11
a
Compound
RLM t1/2
(min)
CYP3A4 IC50
(nM)
CYP2C9 IC50
(nM)
CYP2D6 IC50
(nM)
NHBoc
O
N
7
9a
2
2
>30,000
>30,000
>30,000
3300
7200
4800
22,000
11,400
>30,000
9400
4100
2400
8000
2400
>30,000
23,100
>30,000
20,000
>30,000
>30,000
>30,000
—
11,600
5900
>30,000
11,600
>30,000
25,000
>30,000
16,600
O
X
10f
11e
11g
11h
11i
11j
11k
11l
11m
11n
30
30
47
7
11
>60
>60
46
25
25
a
a
Compound
X
Enzyme IC50 (nM)
Adipocyte IC50 (nM)
9a
H
OH
2.7
3.8
15.2
15.7
66.9
>100
26.5
>100
7.5
10a
10b
10cb
10d
10e
10f
8500
NHAc
15.7
289
5.8
98.0
3.4
10,300
17,100
14,200
18,600
26,600
CO2Me
CH2OH
CMe2OH
CONH2
CONH2
CN
10gc
10h
4.4
7.4
17.6
24.1
a
See Ref. 33.
a
b
c
See Ref. 32.
1:1 E/Z mixture.
(S) isomer of 10f.
as the only trifluoromethylpyridyl compound with t1/2 >60 min. All
the trifluoromethylpyridyl compounds profiled had unacceptable
IC50 values of <10 lM against CYP3A4 with the exception of 11j.
Replacement of the trifluoromethyl substituent in 11e and 11g
with the more polar, but still electron withdrawing, cyano group
gave 11k and 11l, respectively. The cyano group improved adipo-
cyte potency (Table 4), maintained good metabolic stability, and
reduced CYP inhibition (Table 5). The (R) isomers 11k and 11l were
more potent and stable in RLM than the corresponding (S) isomers
11m and 11n (Table 5); thus, the (R) isomers were selected for fur-
ther characterization.
Compounds 11k and 11l had drug-like clog D7.0 and PSA values
(Table 2), and desirable in vitro and in vivo profiles. They experi-
enced <3Â loss in potency when assayed in the presence of 50% hu-
man plasma and exhibited >1000Â selectivity over three other
hydroxysteroid dehydrogenases, including 11b-HSD2 (Table 6).
Despite their stability in RLM, both compounds experienced mod-
In a further effort to improve the cellular potency of 10f, we
reinvestigated substituents on the pyrrolidine ring by replacing
the t-butoxycarbonyl group with heterocycles. A 2-pyrimidinyl
group was initially selected since it recapitulates many of the
hydrogen bond accepting features of the t-butoxycarbonyl group.
Compound 11a, the first analog prepared, suffered a 2Â loss in cel-
lular potency compared to 10f (Table 4); however, the regioiso-
meric 4-pyrimidinyl compound 11b demonstrated potency
equivalent to 10f. Replacement of the electron withdrawing CF3
group in 11b with dimethyl substituents (11c) reduced enzyme
potency by 10Â. These results led us to focus on CF3 substituted
heterocycles.
Replacement of the pyrimidine ring with a CF3 substituted
1,3,4-thiadiazole (11d) reduced potency; however, three of the
four isomeric trifluoromethylpyridyl analogs (11e, 11g and 11h)
had enzyme potency <1 nM and cellular potency <5 nM. Com-
pounds with the (S) stereochemistry on the pyrrolidine ring were
also investigated: 11i and 11j had comparable enzyme and cellular
potency to epimers 11e and 11g. Analogs with cellular IC50 <5 nM
were further profiled for metabolic stability in rat liver microsomes
(RLM) and for inhibition of selected CYP isozymes (Table 5). Half
lives in the presence of RLM varied considerably with 11j emerging
Table 6
In vitro characterization of 11k and 11l
NH
Het
O
N
H2N
O
O
Het
NC
CN
Table 4
Heteroaryl compounds 11
N
N
NH
Het
11k
11l
O
N
Enzyme + plasma IC50
11b-HSD2 IC50
3b-HSD2 IC50
17b-HSD1 IC50
Rat hepatocyte CL
(nM)
2.4
7350
>10,000
>10,000
41
3.2
8650
>10,000
>10,000
33
H2N
O
(nM)
(nM)
(nM)
O
(mL/min/kg)
Compound Het
Stereo Enzyme
Adipocyte
a
a
IC50 (nM) IC50 (nM)
11a
11b
11c
11d
11e
11f
11g
11h
11i
11j
11k
11l
11m
11n
4-CF3-2-pyrimidinyl
2-CF3-4-pyrimidinyl
2,6-diMe-4-pyrimidinyl
R
R
R
R
R
R
R
R
S
5.2
1.9
20.7
21.3
0.55
3.1
0.82
0.67
0.63
0.74
0.87
1.5
16.4
7.0
>100
>100
1.1
14. 7
5.0
1.4
0.5
2.7
0.4
2.7
Table 7
Rat pharmacokinetics of 11k and 11la
5-CF3-2-(1,3,4-thiadiazolyl)
3-CF3-2-pyridyl
4-CF3-2-pyridyl
5-CF3-2-pyridyl
6-CF3-2-pyridyl
3-CF3-2-pyridyl
5-CF3-2-pyridyl
3-CN-2-pyridyl
5-CN-2-pyridyl
3-CN-2-pyridyl
5-CN-2-pyridyl
Het
NC
CN
N
N
11k
11l
S
IV AUC
Vss
MRT disp
IV CL
F
(nM h)
(L/kg)
(h)
(mL/min/kg)
(%)
398
0.70
0.3
42
209
2.2
0.5
80
R
R
S
2.6
1.7
7.6
5.2
S
42
27
a
a
See Ref. 32.
IV dose = 1
lmol/kg, PO dose = 5 lmol/kg.

C. M. Tice et al. / Bioorg. Med. Chem. Lett. 20 (2010) 6725–6729
6729
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Wehave describedthe optimization of 6, a hydrophobicmolecule
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cortisol, which was quantified using SPA beads and a microscintillation plate
reader.32 Biochemical assays used recombinant 11b-HSD1 isolated as
a
microsomal preparation from transfected CHO cells. Assays were performed
in 25 mM HEPES, pH 7.4, 50 mM KCl, 2.5 mM NaCl, 1 mM MgCl2, 1 mM NADPH
and 80 nM [3H]-cortisone at room temperature for 1 h. Cell-based potency was
assessed in differentiated human adipocytes by the addition of cortisone
(80 nM [3H]-cortisone) followed by incubation at 37 °C for 2 h. IC50 values
represent the mean of at least duplicate assays and were generated from an 8-
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33. Rat liver microsome half life (RLM t1/2) was determined as follows. Compounds
were incubated in phosphate buffer with rat liver microsomes containing a
total concentration of CYP isozymes of 250 pmol/mL. At selected time points,
aliquots were quenched and analyzed by LC–MS.