Identification of 2-aminooxazole amides as acyl-CoA: Diacylglycerol acyltransferase 1 (DGAT1) inhibitors through scaffold hopping strategy

Article abstract of DOI:10.1016/j.bmcl.2013.09.048

A scaffold hopping strategy was successfully applied in discovering 2-aminooxazole amides as potent DGAT1 inhibitors for the treatment of dyslipidemia. Further optimization in potency and PK properties resulted in a lead series with oral in vivo efficacy in a mouse postprandial triglyceridemia (PPTG) assay.

Full text of DOI:10.1016/j.bmcl.2013.09.048

Bioorganic & Medicinal Chemistry Letters 23 (2013) 6410–6414
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Identification of 2-aminooxazole amides as acyl-CoA: Diacylglycerol
acyltransferase 1 (DGAT1) inhibitors through scaffold hopping
strategy
Hyunjin M. Kim a, , Michelle D. Smith , Jae-Hun Kim , Mary Ann Caplen , Tin Yau Chan ,
⇑
b
a
a
a
a
b
b
b
Brian A. McKittrick , John A. Cook , Margaret van Heek , Jean Lachowicz
a
Discovery Chemistry, Merck Research Laboratories, 2000 Galloping Hill Road, Kenilworth, NJ 07033, USA
Discovery Biology, Merck Research Laboratories, 2000 Galloping Hill Road, Kenilworth, NJ 07033, USA
b
a r t i c l e i n f o
a b s t r a c t
Article history:
A scaffold hopping strategy was successfully applied in discovering 2-aminooxazole amides as potent
DGAT1 inhibitors for the treatment of dyslipidemia. Further optimization in potency and PK properties
resulted in a lead series with oral in vivo efficacy in a mouse postprandial triglyceridemia (PPTG) assay.
Ó 2013 Elsevier Ltd. All rights reserved.
Received 26 August 2013
Accepted 17 September 2013
Available online 25 September 2013
Keywords:
DGAT1 inhibitors
Treatment for dyslipidemia
Scaffold hopping
Triglycerides are one of the neutral lipids that are utilized to
store free long chain fatty acids. Excess accumulation of triglycer-
ides results in obesity and is associated with insulin resistance.
pocytes.4 Furthermore, on a high fat diet, DGAT1 knockout mice
were resistant to diet-induced obesity, despite normal food intake
and intestinal triglyceride resynthesis. Although these mice had
1
5
Triglycerides in the body are obtained by two major routes, one
through absorption from food sources and the other through bio-
synthesis. Therefore, limiting the absorption of dietary triglycer-
ides and the inhibition of their production in the body would be
effective methods to address the diseases caused by lipid imbal-
ance, such as obesity and type 2 diabetes. In the biosynthesis of tri-
glycerides, the final step of the process is carried out by the
enzyme called diacylglycerol acyltransferases (DGAT) with co-fac-
lower levels of triglycerides, diacylglycerol and fatty acid levels re-
mained similar to the wild type mice. These results strongly sug-
gested that inhibition of DGAT1 would be a beneficial point of
intervention for diseases caused by excessive lipids.
DGAT has been a target of great interest, and many medicinal
chemistry programs focusing on DGAT1 inhibition have been dis-
5
6
–15
closed.
Hoffmann-La Roche reported a class of heterocyclic
as potent DGAT1 inhibitors
amide compounds such as
1
2
16
tor acyl-CoA. DGAT is an enzyme widely expressed at the endo-
(Fig. 1). There are two features of 1 that caught our attention
when we initiated the DGAT1 inhibitor program. One, unlike sev-
eral other DGAT1 inhibitors reported in the literature such as
plasmic reticulum, and catalyzes the fatty acid esterification
reaction on diacylglycerol in the liver and adipose tissue during
the biosynthesis of triglycerides. Additionally, DGAT plays an
important role during triglyceride absorption in the small intestine
1
7–20
2,
1
does not possess the acid functionality, and the
2
where triglyceride resynthesis is the last step of its absorption.
There are two isoforms of DGAT that are involved in triglyceride
synthesis. DGAT2 has been found in a wide range of tissues and is
present in especially high levels in the liver. Knockout of the
DGAT2 gene in mice was lethal due to severe essential fatty acid
O
OH
region A
O
CF
3
N
H
N
O
CF
3
O
N
3
O
O
deficiency. On the other hand, DGAT1 knockout mice were shown
N
N
Me
Me
S
N
H
F
to be viable. Compared to the wild-type mice on a chow diet,
DGAT1 knockout mice had 50% less adipose mass and smaller adi-
1
2
region B
hDGAT1 IC50 85 nM
mDGAT1 IC50 2849 nM
⇑
Corresponding author. Tel.: +1 908 740 3198.
E-mail address: hyunjin.kim@merck.com (H.M. Kim).
Figure 1. Examples of DGAT1 inhibitors.
0
960-894X/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.bmcl.2013.09.048

H. M. Kim et al. / Bioorg. Med. Chem. Lett. 23 (2013) 6410–6414
6411
Table 1
Table 3
Oxazole amide replacements
Saturated heterocycles on the oxazole
H
N
CF
3
CF3
O
R
N
N
H
N
H
N
O
O
R
O
R
O
N
N
O
N
N
N
N
3
OEt
N
OEt
Compd
R
hDGAT-1 IC50 (nM)
1712
5
6
O
O
Compd
R
hDGAT-1 IC50 (nM)
1708
3
3
a
N
5a
N
3
F C
N
S
S
5b
6870
66
b
1616
O
N
5c
O
3
c
1560
2837
5d
N
93
3
d
N
5e
F
4749
F
N
N
Table 2
Region B SAR study of 2-phenyloxazole amides
6a
1508
938
6b
CF
3
N
H
N
O
N
N
6
6
c
79
O
N
R
4
d
228
Compd
R
hDGAT-1 IC50 (nM)
84
N
4a
4b
4c
OEt
O
CF
3
CF3
O
O
O
S
a
N
N
b
F
3
C
OEt
OEt
OEt
H
2
N
O
Br
O
9
N
N
Cl
7
179
474
O
8
O
OEt
CF
3
CF
3
N
N
c
OEt
d
OH
N
O
N
O
N
O
O
1
0
11
O
O
Scheme 1. Reagents and conditions: (a) urea, DMF, 120 °C; (b) t-BuONO, CuBr
CH CN; (c) piperidine,
O.
2
,
N
N
4
4
d
e
N
N
OEt
116
103
3
a,a,a-trifluorotoluene, microwave, 120 °C; (d) NaOH, THF,
H
2
O
O
Cl
H
N
2
O N
2
H N
2
O N
a
b
N
N
N
N
N
Cl
NBoc
NBoc
13
14
1
2
Scheme 2. Reagents and conditions: (a) 1-Boc-piperazine, Hunig’s base, EtOH,
microwave, 140 °C; (b) H , Pd/C, EtOH, EtOAc.
pharmacokinetic properties of 1 were expected to be different
from other acid containing inhibitors. Two, its relatively low
molecular weight (MW 420) would make it a suitable starting
point for further functionalizations in future SAR studies. In our
hands, the compound showed human DGAT1 IC50 of 85 nM, and
mouse DGAT1 IC50 of 2849 nM with tractable PK exemplified by
2
pharmacokinetic properties. In addition, the confirmation of
in vivo efficacy of the new lead series was deemed to be important
prior to the promotion of the program to the lead optimization
stage.
Initially, we focused our efforts on replacing oxazole amides
with different heterocyclic amides in region A. The compounds ob-
tained from this work turned out to be much weaker DGAT1
Rat PO AUC of 12.7
a novel lead scaffold based on this molecule, two regions of 1
region A and B) were explored to obtain suitable potency and
l
M h (6 h).21 For our initial efforts to identify
(

6
412
H. M. Kim et al. / Bioorg. Med. Chem. Lett. 23 (2013) 6410–6414
CF
3
CF
3
N
H
2
N
H
N
N
a
OH
+
N
O
N
O
N
N
O
O
NBoc
N
N
15
1
1
14
NBoc
OEt
CF
3
CF
3
N
N
H
H
b
N
c
N
N
O
N
O
O
O
N
N
N
N
1
6
NH
6a
N
O
2 2
Scheme 3. Reagents and conditions: (a) HATU, Hunig’s base, DMAP, DMF; (b) 4 N HCl/dioxane; (c) ethyl chloroformate, Hunig’s base, CH Cl .
Table 4
Region B SAR study of 2-aminooxazole amides
CF
3
N
H
N
N
O
O
N
R
17
Compd
R
hDGAT1 IC50 (nM)
1508
mDGAT1 IC50 (nM)
N
6
1
1
1
1
1
1
1
1
1
a
N
N
N
N
N
N
N
N
N
N
O
O
O
O
O
O
O
O
O
O
O
O
O
O
N
N
7a
7b
7c
7d
7e
7f
7g
7h
7i
41
79
1205
N
N
N
N
Cl
631
5837
311
1381
731
788
41
1387
F
N
N
N
H
N
H
N
Cl
H
N
26

H. M. Kim et al. / Bioorg. Med. Chem. Lett. 23 (2013) 6410–6414
6413
Table 4 (continued)
Compd
R
hDGAT1 IC50 (nM)
mDGAT1 IC50 (nM)
134
O Cl
N
N
1
7j
N
46
HN
1
7k
3870
180
67
N
N
1
7l
N
N
Cl
1
7m
2269
N
inhibitors (Table 1), suggesting that oxazole amide was the best
moiety in region A to maintain the inhibitory activity.
Then, we shifted our focus on developing SAR in region B. We
discovered that 4-substituted piperidine and piperazine produced
quite potent compounds (Table 2). Ethyl piperidine-4-carboxylate
derivative 4a possessed similar IC50 as 1 while homologated ester
(Scheme 3). Further functionalization of 15 was carried out
by deprotection of the Boc group followed by carbamate forma-
tion to yield 6a. Again, several other reactions including urea
formations and sulfonamide formations were also carried out
on 16 in the preparation of other compounds described in this
communication.
4
b was slightly less potent. Methyl sulfonamide 4c displayed even
To further probe the SAR of this new scaffold, we decided to
have region A fixed as piperidinyloxazole as in 6a and began re-
exploring region B (Table 4). Among the piperazinyl carbamate
compounds (6a, 17a, 17b, and 17c), branched alkyl carbamates
17a and 17b turned out to be more potent in the series (hDGAT1
IC50 41 nM and 79 nM, respectively). In general, less inhibitory
activities were obtained for piperazinyl amide compounds (17d,
17e, and 17f), although SAR trends similar to the piperazinyl carba-
mate series were observed. Contrary to the carbamate and amide
cases, piperazinyl alkyl ureas did not follow a similar trend in
SAR. Thus, branched alkyl ureas 17g and 17h were less active,
while aryl urea 17i was one of the most potent in the series
(hDGAT1 IC50 41 nM). Homopiperazinyl urea 17j was also quite ac-
tive and demonstrated flexibility of the region to accommodate
some small changes. In addition, several aryl piperidine and aryl
piperazine compounds were prepared (17k, 17l, and 17m). In gen-
eral, aryl piperazine compounds exhibited better potencies than
aryl piperidine compounds and aryl piperazine 17m showed the
best potency (hDGAT1 IC50 67 nM) among these types. For selected
cases, mouse DGAT1 potency was measured as well and 17a and
17m showed some selective inhibition favoring human DGAT1
over mouse DGAT1.
less inhibitory activity. However, ethyl carbamate 4d and urea ana-
log 4e showed potent inhibition of human DGAT1 enzymatic
activities.
With newly gained insights on SAR trends in region B, we began
to explore possible modifications in region A to further improve
potency and PK. After extensive efforts to find suitable modifica-
tions, we discovered that the C2-phenyl on the oxazole could be re-
placed with saturated heterocycles such as piperidine without loss
of inhibitory activity (Table 3). Replacement of the phenyl group
with pyrrolidine (compound 5a) or morpholine (compound 5b)
showed significant loss of potency (hDGAT1 IC50 1708 nM and
6
870 nM, respectively). However, piperidine replacement was
more successful, producing potent compounds 5c and 5d (hDGAT1
IC50 66 nM and 93 nM, respectively). As noted in the case of 5b and
5
e, not all six member saturated ring compounds were active and
future SAR optimization potentials were observed. For substituted
piperidines, 2-, 3-, and 4-methylpiperidine analogs 6b, 6c, and 6d
were prepared and compared with unsubstituted piperidine analog
6
a. Parallel to the case of 5c and 5d, 6a and 6b were similar in po-
tency (hDGAT1 IC50 1508 nM and 938 nM, respectively). Further
improvement of the potency was observed for 6c and 6d with 3-
methylpiperidinyloxazole 6c being the most potent (hDGAT1 IC50
As shown for the representative compounds 17a and 17i
(Table 5), PK properties of this series of compounds were quite
acceptable at the hit to lead stage. These compounds have high
Rat AUC and good solubility. As for other properties, their clogP
7
9 nM).
The synthesis of the representative compound 6a commenced
from the combination of ethyl 4,4,4-trifluoro-2-chloroacetoacetate
2
2
23
(7) and urea, which resulted in the formation of 8 (Scheme 1).
values were in the acceptable range (clogP < 5) , even though
Modified Sandmeyer conditions were utilized to convert 8 to 9,
which was a key intermediate for the syntheses of several
molecular weights are slightly higher than desired. The Cyp P450
profiles of the series were also acceptable and so were their hERG
2
-aminooxazole compounds. In the particular case of 6a, 9 was
inhibitions. On the other hand, human hepatocyte clearance for
6
treated with piperidine to provide 10, which was successfully con-
verted to 11 upon basic hydrolysis.
these compounds were high (24.2
l
L/min/10 cells for 17a and
6
16.9 lL/min/10 cells for 17i). Another potential area for improve-
The preparation of the other key intermediate 14 was initiated
by treatment of 2-chloro-5-nitropyridine (12) with 1-Boc
piperazine to provide 13 (Scheme 2). Alternatively, other amines
in place of 1-Boc piperazine were successfully employed for the
syntheses of other compounds in this study. The nitro group of
ment was plasma protein binding as both compounds exhibited
greater than 99% binding. Overall, the 2-aminooxazole amide
series has been shown to possess suitable PK properties as a lead
series.
Selected compounds from this series were tested in a mouse
postprandial triglyceridemia (PPTG) assay to determine in vivo
1
3 was reduced to give aminopyridine 14.
2
4,25
HATU mediated coupling of acid 11 and aminopyridine 14
activity (Table 6).
The compounds were orally dosed (3 mg/
in the presence of catalytic amount DMAP provided 15
kg or 10 mg/kg) to mice fasted overnight and treated with an oral

6
414
H. M. Kim et al. / Bioorg. Med. Chem. Lett. 23 (2013) 6410–6414
Table 5
PK for representative compounds
1
7a
511
3.87
17i
Mol Wt
clogP
578
3.95
Solubility
100
69
24.2
l
M
10
25
16.9
l
M
Rat AUC (
hHepatocyte Cl (
P450 IC50 (pre/co
lM h)
l
l
L/min/106 cells)
M)
3A4 (>20/>20), 2D6 (>20/>20), 2C9 (>20/>20)
3A4 (5.0/>20), 2D6 (>20/>20), 2C9 (9.7/6.4)
hERG ionwork (% inhib. at 10
Plasma protein binding
lM)
40%
99.6%
99.9%
Judd, A. S.; King, A. J.; Klix, R. C.; Larson, K. J.; Lau, Y. Y.; Marsh, K. C.;
Mittelstadt, S. W.; Plata, D.; Rozema, M. J.; Segreti, J. A.; Stoner, E. J.; Voorbach,
M. J.; Wang, X.; Xin, X.; Zhao, G.; Collins, C. A.; Cox, B. F.; Reilly, R. M.; Kym, P.
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Table 6
Reduction of plasma triglyceride level
%
1
0 mg/kg
3 mg/kg
1
1
2. Bali, U.; Barba, O.; Dawson, G.; Gattrell, W. T.; Horswill, J. G.; Pan, D. A.; Procter,
M. J.; Rasamison, C. M.; Sambrook-Smith, C. P.; Taylor-Warne, A.; Wong-Kai-In,
P. Bioorg. Med. Chem. Lett. 2012, 22, 824.
1
1
1
7a
7b
7i
À12
À27
À63
3. Mougenot, P.; Namane, C.; Fett, E.; Camy, F.; Dadji-Faihun, R.; Langot, G.;
Monseau, C.; Onofri, B.; Pacquet, F.; Pascal, C.; Crespin, O.; Ben-Hassine, M.;
Ragot, J.-L.; Van-Pham, T.; Philippo, C.; Chatelain-Egger, F.; Peron, P.; Le Bail, J.-
C.; Guillot, E.; Chamiot-Clerc, P.; Chabanaud, M.-A.; Pruniaux, M.-P.; Schmidt,
F.; Venier, O.; Nicolai, E.; Viviani, F. Bioorg. Med. Chem. Lett. 2012, 22, 2497.
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Jenner, C.; Kemmitt, P. D.; Leach, A. G.; Moody, G. C.; Gutierrez, P. M.;
Newcombe, N. J.; Nowak, T.; Packer, M. J.; Plowright, A. T.; Revill, J.; Schofield,
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À28
bolus of corn oil 15 min after compound dosing. After 2 h of the
corn oil bolus, blood samples were obtained and plasma triglycer-
ide levels were measured. Then the values were compared with
those of the control mice. Three compounds showed activity at
1
1
3
0 mg/kg. And the most potent 17i also showed good activity at
mg/kg. Having demonstrated significant reductions in plasma tri-
1
5. Goldberg, F. W.; Birch, A. M.; Leach, A. G.; Groombridge, S. D.; Snelson, W. L.;
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glyceride levels, the 2-aminooxazole amide series provided a suit-
able lead series, which warrant further lead optimization studies.
In conclusion, we successfully identified the 2-aminooxazole
amide series as the lead for a DGAT1 inhibitor program for the
treatment of diabetes and obesity. From the SAR studies, we were
able to achieve the desired level of in vitro potency. Several com-
pounds in the series have been shown to possess desirable PK
properties. Furthermore, we were able to demonstrate in vivo
activity in reducing plasma triglyceride levels in mice challenged
with a triglyceride load. Thus, with identification of the 2-amino-
oxazole amide series as the lead, the DGAT1 inhibitor hit-to-lead
program has been successfully concluded and transferred to lead
optimization.
2
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2
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1
8
9
.
.
2
011, 54, 2433.
1326). Absorbance values were determined in an automated microplate reader
1
1
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at a wavelength of 600 nm.
2
5. All animal studies were reviewed and approved by the Merck IACUC. The Guide
for the Care and Use of Laboratory Animals was followed in the conduct of the
animal studies. Veterinary care was given to any animals requiring medical
attention.