4-Bicyclic heteroaryl-piperidine derivatives as potent, orally bioavailable Stearoyl-CoA desaturase-1 (SCD1) inhibitors. Part 1: Urea-based analogs

Article abstract of DOI:10.1016/j.bmcl.2013.09.096

A new series of urea-based, 4-bicyclic heteroaryl-piperidine derivatives as potent SCD1 inhibitors is described. The structure-activity relationships focused on bicyclic heteroarenes and aminothiazole-urea portions are discussed. A trend of dose-dependent decrease in body weight gain in diet-induced obese (DIO) mice is also demonstrated.

Full text of DOI:10.1016/j.bmcl.2013.09.096

Bioorganic & Medicinal Chemistry Letters 23 (2013) 6773–6776
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
4-Bicyclic heteroaryl-piperidine derivatives as potent, orally
bioavailable Stearoyl-CoA desaturase-1 (SCD1) inhibitors. Part 1:
Urea-based analogs
⇑
Shyh-Ming Yang , Yuting Tang, Rui Zhang, Huajun Lu, Gee-Hong Kuo, Michael D. Gaul, Yaxin Li,
George Ho, James G. Conway, Yin Liang, James M. Lenhard, Keith T. Demarest, William V. Murray
Cardiovascular and Metabolism Research, Janssen Research and Development, LLC, Welsh & McKean Roads, Spring House, PA 19477, USA
a r t i c l e i n f o
a b s t r a c t
Article history:
A new series of urea-based, 4-bicyclic heteroaryl-piperidine derivatives as potent SCD1 inhibitors is
described. The structure–activity relationships focused on bicyclic heteroarenes and aminothiazole–urea
portions are discussed. A trend of dose-dependent decrease in body weight gain in diet-induced obese
(DIO) mice is also demonstrated.
Received 9 August 2013
Revised 25 September 2013
Accepted 30 September 2013
Available online 9 October 2013
Ó 2013 Elsevier Ltd. All rights reserved.
Keywords:
Stearoyl-CoA desaturase
SCD1 inhibitors
Desaturation index
Obesity
H
2N
O
Stearoyl-CoA desaturase-1 (SCD1) is a critical microsomal en-
zyme that catalyzes the conversion of saturated fatty acid-CoAs
to mono-unsaturated fatty acid-CoAs at C-9 position.1 These
mono-unsaturated fatty acid-CoAs, such as palmitoleic (C16)
R
O
R1
NO2
R1
7
H
N
NH
a
N
R2
O
6
3, 4, 5
Cl
O
O
H
N
N
N
NHMe
H
b
N
N
5b−5e
5f−5i
or
N
O
F
CO2R
n
or c
O
S
1, mSCD1: 4 nM; hSCD1: 37 nM
R1
8, n= 0-1, R= Me or Et
H
N
N
R2
O
3−5
: Bicyclic heteroaryl
CF3
O
N
H
d
N
R1
N
N
N
5m−5n
F
N
O
or e
S
O
S
N
NH2
X
9, X= CH-OTBS or N-Boc
2 (MF-152), rSCD1: 106 nM
Scheme 1. R1 represents bicyclic heteroaryl, such as indole, indoline, benzoisox-
azole, etc. Reagents and conditions: (a) compound 7, Et3N, CH2Cl2, rt, 55–90%, or
R2NH2, triphosgene, Et3N, CH2Cl2, rt, 40–92%; (b) DIBAL-H, CH2Cl2, rt, 5bÀ5e (45–
78%); (c) MeNH2(aq) or NH3(aq), EtOH, 40–50 °C, 5f–5i (59–97%); (d) for X = N-Boc,
TFA, CH2Cl2, rt, 5m (90%); (e) for X = CH-OTBS, TBAF, THF, rt, 5n (92%).
Figure 1. Representative piperidine- and piperazine-derived SCD1 inhibitors.
E-mail address: shyhyang99@yahoo.com (S.-M. Yang).
0960-894X/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.bmcl.2013.09.096

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S.-M. Yang et al. / Bioorg. Med. Chem. Lett. 23 (2013) 6773–6776
acid-CoA and oleic (C18) acid-CoA, are major building blocks in
biosynthesis of lipids including phospholipids, triglycerides,
cholesterol esters and wax esters.2 Four SCD isoforms (SCD1–4)
in rodents and two human genes (SCD1 and SCD5, also known as
SCD2) have been identified and characterized. SCD1,3 with ca.
85% identity across species,4 is abundantly expressed in liver and
adipose tissue and is regulated by several nutritional and hormonal
factors, such as insulin, cholesterol, and poly-unsaturated fatty
acids.5 Moreover, SCD1 deficiency in mice, either naturally
deficient Asebia mice6 or laboratory-created SCD1 knockout
(SCD1À/À) mice,7 has been shown to reduce body adiposity,
increase insulin sensitivity, and impart resistance to diet-induced
obesity.8 The SCD1À/À mice also have lower levels of hepatic
cholesterol esters and triglycerides.9 These observations suggest
inhibition of SCD1 activity may serve as a potential treatment for
obesity, type-II diabetes, and other related metabolic disorders.
In recent years, piperidine and piperazine derived analogs,
exemplified by 110c and 210d (MF-152), have been the major focus
in the design of SCD1 inhibitors10,11 (Fig. 1). These novel skeletons
consist of a 4-phenoxy or 4-phenylcarbonyl motif have been pro-
ven to be potent inhibitors in vitro and efficacious in chronic
in vivo studies. Furthermore, the ortho-substitution of phenyl ring,
such as Cl or CF3, is important to improving potency to low nano-
molar range. In our medicinal chemistry program toward novel
SCD1 inhibitors, we envisioned a ring constrained strategy by teth-
ering the key ortho-substituted carbon to either the ether oxygen
position or carbonyl group position (Fig. 1). The newly designed
4-bicyclic heteroaryl-piperidine templates open avenues to iden-
tify potential inhibitors by exploration of the right-hand portion
of these molecules. Herein we report the initial discovery of
urea-based 4-bicyclic heteroaryl-piperidine analogs (3–5) as po-
tent and orally bioavailable SCD1 inhibitors.
The synthesis of these urea-based analogs 3–5 is straightfor-
ward and shown in Scheme 1. Activated-nitrophenyl carbamate
intermediate 7 was rapidly available by treating the corresponding
R2NH2 with 4-nitrophenyl chloroformate. Treatment of 7 with 4-
bicyclic heteroaryl-piperidine (6)12 afforded target compounds 3–
5 in good yield. Alternatively, compounds 3–5 could be obtained
in a one-pot manner by activation of R2NH2 with triphosgene fol-
lowed by addition of piperidine 6. Elaboration of the ester group
of urea intermediate 8 provided analogs 5b–5e and 5f–5i with an
amide or hydroxyalkyl functionality. Further deprotection of 9
using TFA (X = N-Boc) or TBAF (X = CH-OTBS) gave 5m or 5n,
respectively.
Compound activity was initially measured against rat SCD1 en-
zyme (rSCD1).13 The results are compiled in Table 1 focusing on
structure–activity relationships (SAR) at the bicyclic heteroaryl
portion (R1). With N-methyl benzamide substitution at urea por-
tion, the 6,5-bicyclic rings, such as indole (3a) or indoline (3b),
showed good potency below 200 nM. However, the reversed 3-
substituted indole (3c) or 2-indolone (3d) resulted in a significant
decrease in potency. Substituting the right hand portion of the urea
with a 4-methylthiazole group led to a similar SAR that favored
6,5-ring system (4b vs 4c). A dramatic improvement in potency
to low nanomolar range was achieved by introducing a fluoro-sub-
stitution at the 6-position of the indoline (4d, 26 nM). Additionally,
a 5-fluorobenzoisoxazole substituent (4e) provided good inhibi-
tory activity at 140 nM. The cellular activity against human epithe-
lial carcinoma A431 cells (using 13C-palmitic acid and LC–MS
method)14 was added for further evaluation. In A431 assay, com-
pounds 4d and 4e also exhibited good inhibitory activity at 21
and 54 nM, respectively. These results clearly illustrate the bicyclic
Table 1
SAR of bicyclic heteroaryl substituent R1
R1
R1
O
H
N
H
N
N
N
Me
N
N
H
Me
O
S
O
3
4
a
Compds
R1
rSCD1 IC50 (nM)
3a
170
N
3b
3c
151
N
H
N
Table 2
SAR of urea portion R2
15%b
N
H
O
N
N
R2
3d
4a
4b
7%b
538
153
F
N
N
N
O
5
a
Compds
R2
rSCD1 IC50 (nM)
A431b (nM)
21
N
N
Me
4d
26
S
S
5a
5b
5c
5d
82
52
88
20
108
42
Me
4c
4d
4e
491
26
N
N
N
S
S
OH
N
O
F
F
154
18
OH
N
140
N
S
OH
a
Single experiment or means of at least two runs.
b
Inhibition percentage at 1 lM.

S.-M. Yang et al. / Bioorg. Med. Chem. Lett. 23 (2013) 6773–6776
6775
3
Table 2 (continued)
+
2.06 0.33
a
Compds
R2
rSCD1 IC50 (nM)
A431b (nM)
6
2
1
0
N
+
0.00 0.36
*
5e
8
S
+
0.01 0.24
*
OH
O
+
-0.69 0.38
*
N
N
5f
682
—
S
S
NH2
-
1
30 mpk
bid
10 mpk
qd
30 mpk
qd
-
2
Veh
5g
5h
5i
22
49
—
NH2
O
N
Figure 2. Dose-dependent effect of 4e on BW change in DIO C57BL/6 mice
(mean se, ⁄P <0.05 vs vehicle-treated group, results were summarized from three
experiments).
O
20%c
93
S
S
NHMe
N
With suitable bicyclic heteroaryl moieties identified, we turned
our attention to explore the SAR around the urea portion (R2) by
focusing the substituent effects on the thiazole ring. In general, a
variety of substitutions such as methyl (4d and 5a) or hydroxyalkyl
(5b–5e) at the 4- or 5-position of the thiazole were well tolerated
without significant loss of potency in both rSCD1 and A431 assay
(Table 2). Specifically, the 4-methyl- and 4-hydroxymethyl-substi-
tuted thiazoles were about two to three fold more potent than the
5-substituted thiazole analogs (4d and 5b vs 5a and 5c). The elon-
gated hydroxyalkyl analogs (5d–5e) appeared to have slightly bet-
ter inhibitory activity. However, attaching a primary amide at the
4-position (5f, 682 nM) was particularly disfavored that caused a
31-fold loss of potency as compared to the 5-substituted primary
amide analog 5g (22 nM). A similar trend that favored 5-substitu-
tion on the thiazole ring was observed in the methyl amide-substi-
tuted analogs 5h–5i, though they were less potent than the
corresponding primary amides 5f–5g.
206
NHMe
O
N
5j
347
204
76
17
S
S
N
N
5k
O
N
S
5l
46%c
—
Me
O
N
N
5m
5n
29
14
16
1
S
S
Based on the encouraging results observed with compounds 4d
and 5a–5e that tolerated well for substitution at both 4- and 5-po-
sition, a series of di-substituted bicyclic thiazoles were next syn-
thesized and evaluated (Table 2). Compounds 5j–5l achieved
NH
modest potency (204 nM to ca. 1 lM) presumably due to the loss
of hydrogen bonding donor capability. In fact, adding NH (5m)
and OH (5n) groups gained the potency back to 14–29 nM with im-
proved inhibitory activity against A431 cell (1–16 nM). A compar-
OH
N
S
ison between analogs 5l (ca. 1 lM) and 5o (96 nM) clearly
5o
5p
96
18
113
30
N
OH
demonstrated again the critical H-bonding interaction for SCD1 en-
zyme potency. Finally, a bicyclic pyrazole amide (5p) was found to
be very potent against both rSCD1 enzyme and A431 cell (18 nM
and 30 nM, respectively).
Among the compounds tested in vitro, analog 4e was first se-
lected for further evaluation. The pharmacokinetic (PK) profiles
in mouse and rat are summarized in Table 3. The results reveal that
4e has a low clearance level (CL: 0.5 mL/min/kg), high plasma con-
centration, and good bioavailability in rat (F%: 36.9%). An excellent
O
N
N
NH
O
a
b
c
Single experiment or means of at least two runs.
Using 13C-palmitic acid and LC–MS method, see Ref. 14.
Inhibition percentage at 1 lM.
exposure of 4e in diet-induced obese (DIO) C57BL/6 mice (Cmax
:
14.9 g/mL) was also obtained. With
l
g/mL; AUC24 h: 169.3 h ꢀ
l
the favorable PK profile, 4e was further evaluated for in vivo effi-
heteroaryl-substituted piperidine is a suitable alternative to the
original phenoxy-piperidine design.
cacy in DIO C57BL/6 mice at 10 mg/kg q.d. (once-daily), 30 mg/kg
Table 3
The PK profiles of 4e in DIO mice and rats
b
Species
Administrationa
t1/2 (h)
Cmax
(l
g/mL)
AUC24 h (h ꢀ
49.8
118.5
169.3
lg/mL)
Vss (L/kg)
CL (mL/min/kg)
0.5
F%
Rat
Rat
DIO mice
iv (2 mg/kg)
po (10 mg/kg)
po (30 mg/kg)
3.8
6.3
3.1
9.2
8.8
14.9
0.19
36.9
—
a
n = 4 for rat; n = 3 for DIO mice.
Cmax = Co (t = 0) for iv administration.
b

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S.-M. Yang et al. / Bioorg. Med. Chem. Lett. 23 (2013) 6773–6776
Xu, L.; Zhang, L.; Oballa, R. Bioorg. Med. Chem. Lett. 2011, 21, 479; For human
Table 4
SCD1-specific SCD inhibitors, see: (h) Powell, D. A.; Ramtohul, Y.
Lebrun, M.-E.; Oballa, R.; Bhat, S.; Falgueyret, J.-P.; Guiral, S.; Huang, Z.;
Skorey, K.; Tawa, P.; Zhang, L. Bioorg. Med. Chem. Lett. 2010, 20, 6366.
11. For liver-targeted SCD1 inhibitors, see: (a) Oballa, R. M.; Belair, L.; Black, W. C.;
Bleasby, K.; Chan, C. C.; Desroches, C.; Du, X.; Gordon, R.; Guay, J.; Guiral, S.;
Hafey, M. J.; Hamelin, E.; Huang, Z.; Kennedy, B.; Lachance, N.; Landry, F.; Li, C.
S.; Mancini, J.; Normandin, D.; Pocai, A.; Powell, D. A.; Ramtohul, Y. K.; Skorey,
K.; Sorensen, D.; Sturkenboom, W.; Styhler, A.; Waddleton, D. M.; Wang, H.;
Wong, S.; Xu, L.; Zhang, L. J. Med. Chem. 2011, 54, 5082; (b) Koltun, D. O.;
Zilbershtein, T. M.; Migulin, V. A.; Vasilevich, N. I.; Parkhill, E. Q.; Glushkov, A.
I.; McGregor, M. J.; Brunn, S. A.; Chu, N.; Hao, J.; Mollova, N.; Leung, K.;
Chisholm, J. W.; Zablocki, J. Bioorg. Med. Chem. Lett. 2009, 19, 4070; (c) Koltun,
D. O.; Vasilevich, N. I.; Parkhill, E. Q.; Glushkov, A. I.; Zilbershtein, T. M.;
Mayboroda, E. I.; Boze, M. A.; Cole, A. G.; Henderson, I.; Zautke, N. A.; Brunn, S.
A.; Chu, N.; Hao, J.; Mollova, N.; Leung, K.; Chisholm, J. W.; Zablocki, J. Bioorg.
Med. Chem. Lett. 2009, 19, 3050; (d) Leclerc, J. P.; Falgueyret, J.-P.; Girardin, M.;
Guay, J.; Guiral, S.; Huang, Z.; Li, C. S.; Oballa, R.; Ramtohul, Y. K.; Skorey, K.;
Tawa, P.; Wang, H.; Zhang, L. Bioorg. Med. Chem. Lett. 2011, 21, 6505; (e) Powell,
D. A.; Black, W. C.; Bleasby, K.; Chan, C.-C.; Deschenes, D.; Gagnon, M.; Gordon,
R.; Guay, J.; Guiral, S.; Hafey, M. J.; Huang, Z.; Isabel, E.; Leblanc, Y.; Styhler, A.;
Xu, L.-J.; Zhang, L.; Oballa, R. M. Bioorg. Med. Chem. Lett. 2011, 21, 7281; (f)
Ramtohul, Y. K.; Powell, D.; Leclerc, J.-P.; Leger, S.; Oballa, R.; Black, C.; Isabel,
E.; Li, C. S.; Crane, S.; Robichaud, J.; Guay, J.; Guiral, S.; Zhang, L.; Huang, Z.
Bioorg. Med. Chem. Lett. 2011, 21, 5692; (g) Lachance, N.; Guiral, S.; Huang, Z.;
Leclerc, J.-P.; Li, C. S.; Oballa, R. M.; Ramtohul, Y. K.; Wang, H.; Wu, J.; Zhang, L.
Bioorg. Med. Chem. Lett. 2012, 22, 623; (h) Lachance, N.; Gareau, Y.; Guiral, S.;
Huang, Z.; Isabel, E.; Leclerc, J.-P.; Léger, S.; Martins, E.; Nadeau, C.; Oballa, R.
M.; Ouellet, S. G.; Powell, D. A.; Ramtohul, Y. K.; Tranmer, G. K.; Trinh, T.; Wang,
H.; Zhang, L. Bioorg. Med. Chem. Lett. 2012, 22, 980; (i) Deng, Y.; Yang, Z.; Shipps,
G. W., Jr.; Lo, S.-M.; West, R.; Hwa, J.; Zheng, S.; Farley, C.; Lachowicz, J.; Heek,
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Lett. 2013, 23, 791.
Plasma liver enzyme levels after 10 days treatment in DIO mice
Treatment
ALT (U/L)
AST (U/L)
Vehicle
58
57
68 12
41
5
7
99
89
93
9
7
7
4e, 10 mg/kg q.d.
4e, 30 mg/kg q.d.
4e, 30 mg/kg b.i.d.
3
102 17
q.d., and 30 mg/kg b.i.d. (twice-daily) for 10 days (vehicle, 0.5%
hypromellose).15 As shown in Figure 2, 4e displayed a trend of
dose-dependent decrease in body weight (BW) gain in DIO
C57BL/6 mice. In addition, plasma liver enzyme levels (ALT and
AST) measured at the end of the study were not elevated in drug
treated groups, indicating no potential overt liver toxicity (Table 4).
In summary, a new series of piperidine-urea analogs as potent
SCD1 inhibitors have been discovered. Structurally, the bicyclic
heteroaryl, such as 6-fluoroindoline, substituted piperidine dem-
onstrated a viable alternative to the original phenoxy-piperidine
motif. The selected analog 4e with a favorable PK profile demon-
strated a trend of dose-dependent in vivo efficacy in DIO C57BL/6
mice. Further exploration using the bicyclic heteroaryl scaffold to
identify analogs with improved potency and efficacy will be re-
ported in due course.
Acknowledgment
12. Compound 6 are either commercially available or can be prepared according to
literature procedures. Representative synthetic procedures for 3–5 are
available in Supplementary data.
The authors thank the ADME team for bioanalysis support.
13. Rat liver microsome preparation and SCD1 enzymatic assay: Sprague Dawley rats
from Charles River (200–225 g) underwent 2-cycle of fast-re-feeding (24-h fast
followed by 24-h feeding with fat-free high carbohydrate diet, Research Diet,
D00042802). Liver were removed and homogenized 1:4 (w/v) with pre-chilled
homogenizing buffer (10 mM Tris–HCl, 0.25 M Sucrose, 1 mM EDTA, pH 7.4,
supplemented with protease inhibitor cocktail). The homogenate was first
spun at 12,000g for 15 min at 4 °C, followed by re-centrifugation of the
supernatant at 100,000g for 60 min at 4 °C. The pellet was re-suspended in
2 mL pre-chilled 0.2 mM potassium phosphate buffer, pH 7.2, 10 mM EDTA at
concentration of 20 mg/mL. The RLM preparation was stored at À80 °C. SCD1
Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.bmcl.2013.
09.096.
enzymatic assay was done in a volume of 50 lL with 10 lg RLM in 96-well
polypropylene plate (enzyme reaction buffer contains 0.1 M K-Phosphate
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[3H2O] generated was used as indicator of SCD1 activity.
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20
M U-13C16 palmitic acid followed by addition of 5
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extract fatty acids followed by centrifugation at 3000 rpm for 10 min. Transfer
l
L of Assay Buffer was then added to each well including
l
lL of vehicle or Scd1
l
l
l
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l
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S.; Gordon, R.; Guay, J.; Guiral, S.; Huang, Z.; Robichaud, J.; Skorey, K.; Tawa, P.;
l
l
75 lL to new glass vial in 96-well cluster plate and inject 5 lL for LC/MS
analysis for detection of 13C-palmitic acid and 13C-palmitoleic acid. The ratio of
13C-palmitoleic acid to 13C-palmitic acid was determined as index of SCD1
activity.
15. DIO C57BL/6 mice were orally dosed with either vehicle (0.5%
hypromellose) or compound 4e at 8 AM and 6 PM daily at 10 mL/kg.
Dosing was performed as following: vehicle-treated group (8 AM and 6 PM
with vehicle (0.5% hypromellose)); 10 mg/kg-treated q.d. group (8 AM with
vehicle (0.5% hypromellose) and 6 PM with compound 4e (dosing solution:
1 mg/mL)); 30 mg/kg-treated q.d. group (8 AM with vehicle (0.5%
hypromellose) and 6 PM with compound 4e (dosing solution: 3 mg/mL));
30 mg/kg-treated b.i.d. group (8 AM and 6 PM with compound 4e (dosing
solution: 3 mg/mL)).