Phenoxyacetic acids as PPARδ partial agonists: Synthesis, optimization, and in vivo efficacy

Article abstract of DOI:10.1016/j.bmcl.2011.02.077

A series of phenoxyacetic acids as subtype selective and potent hPPARδ partial agonists is described. Many analogues were readily accessible via a single solution-phase synthetic route which resulted in the rapid identification of key structure-activity r

Full text of DOI:10.1016/j.bmcl.2011.02.077

Bioorganic & Medicinal Chemistry Letters 21 (2011) 2345–2350
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Phenoxyacetic acids as PPARd partial agonists: Synthesis, optimization,
and in vivo efficacy
Karen A. Evans a, , Barry G. Shearer b, David D. Wisnoski a, Dongchuan Shi a, Steven M. Sparks b,
⇑
Daniel D. Sternbach b, Deborah A. Winegar b, Andrew N. Billin b, Christy Britt b, James M. Way b,
Andrea H. Epperly b, Lisa M. Leesnitzer c, Raymond V. Merrihew c, Robert X. Xu c,
Millard H. Lambert c, Jian Jin a
a Discovery Medicinal Chemistry, Discovery Research, GlaxoSmithKline Pharmaceuticals, 1250 South Collegeville Road, PO Box 5089, Collegeville, PA 19426-0989, United States
b Metabolic Diseases Center of Excellence for Drug Discovery, GlaxoSmithKline Pharmaceuticals, Five Moore Drive, PO Box 13398, Research Triangle Park, NC 27709-3398,
United States
c Discovery Research, GlaxoSmithKline Pharmaceuticals, Five Moore Drive, PO Box 13398, Research Triangle Park, NC 27709-3398, United States
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of phenoxyacetic acids as subtype selective and potent hPPARd partial agonists is described.
Many analogues were readily accessible via a single solution-phase synthetic route which resulted in
the rapid identification of key structure–activity relationships (SAR), and the discovery of two potent
exemplars which were further evaluated in vivo. Details of the SAR, optimization, and in vivo efficacy
of this series are presented herein.
Received 29 November 2010
Revised 18 February 2011
Accepted 22 February 2011
Available online 15 March 2011
Ó 2011 Elsevier Ltd. All rights reserved.
Keywords:
PPARd
Partial agonist
The peroxisome proliferator activated receptors (PPARs) are
important members of the nuclear receptor superfamily. These
receptors are ligand activated transcription factors known to play
a key role in the catabolism and storage of dietary fats. Three
potential templates to exploit. The goal of this effort was to iden-
tify potent and selective partial PPARd agonists for testing across
a panel of in vivo models. Standard PPAR binding and cell-based re-
porter assays were used as primary screens to profile compounds.
receptor subtypes, PPAR
a, PPARc, and PPARd, exhibiting distinct
Phenoxyacetic acid 2 (hPPARd binding pIC50 = 5.9, hPPARd
tissue expressions have been identified, and represent attractive
therapeutic targets with promising clinical potential.1 PPARd re-
mains the least understood PPAR subtype and currently no mar-
keted PPARd agonists exist. However, evidence implicating PPARd
as a key regulator of lipid homeostasis and glucose disposal is
growing.2 For example, the full PPARd agonist GW5015163 (1)
has been shown to reduce serum triglycerides and prevent the de-
crease of HDL-c and apoA-1 levels observed in sedentary human
volunteers.2c These positive results suggest that PPARd is a promis-
ing target for the novel treatment of metabolic diseases.
pEC50 = 6.8, 75%max) was identified as a compound with modest
potency and submaximal efficacy that could serve as a promising
starting point for SAR exploration.5
A number of structural modifications of agonist 2 were initially
designed and implemented. This ultimately resulted in the prepa-
ration of compound 3 (Fig. 1) which was in fact a potent and selec-
tive partial agonist of PPARd (hPPARd binding pIC50 d = 7.3; hPPARd
functional pEC50 = 7.6, 56%max). Compound 3 also demonstrated
promising pharmacokinetic properties in the mouse via oral
administration (DNAUC0–24 h = 575 ng h/mL/mg/kg).6 This com-
pound became our new partial agonist lead compound in order
to more fully explore the structure–activity relationships of this
new series, as well as to further optimize for potency, oral expo-
sure, and in vivo efficacy.
While the number of PPARd selective agonists described in the
literature has grown in recent years, additional chemical tool com-
pounds with a range of functional activity profiles are still needed
to further elucidate the biologic roles of PPARd and provide addi-
tional insights into potential therapeutic utilities of modulating
this receptor.
Systematic structural modifications of this template at five
positions of diversity were primarily achieved through a solu-
tion-phase synthesis as described in Scheme 1. The appropriate
As part of our effort to identify PPARd partial agonists,4 we
searched our internal database of PPAR program compounds for
phenol
4 was treated with substituted bromoacetate 5 and
potassium carbonate in N,N-dimethylformamide (DMF) under
microwave conditions to afford the phenoxyacetic acid ester 6.
Immediately following treatment of 6 with chlorosulfonic acid,
⇑
Corresponding author. Tel.: +1 610 917 7764; fax: +1 610 917 6151.
E-mail address: karen.a.evans@gsk.com (K.A. Evans).
0960-894X/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bmcl.2011.02.077

2346
K. A. Evans et al. / Bioorg. Med. Chem. Lett. 21 (2011) 2345–2350
O
O
O
CF3
CF3
O
HO
HO
S
N
S
S
N
O
O
1 (GW501516)
2
O
Cl
O
HO
N
S
O
O
3
hPPAR Binding pIC50
hPPAR δReporter
pEC50
Compound
%Max
α
γ
δ
5.7
4.8
4.7
5.2c
5.3
5.1
8.3
5.9
7.3
8.5c
98
75
56
1
2b
6.8
7.6c
3
Figure 1. Structures and assay results for lead phenoxyacetic acids. Values are means of at least two experiments, standard deviation (SD) <0.2 unless otherwise noted.
bValues are n = 1. cSD = 0.3.
1
MeO2C
(a)
R
R2
O
R2
R2
O
5
(b)
Br
O
HO
2N
R2
O
R2
R2
S
O
O
+
1
R
R1
Cl
4
6
7
O
O
O
R2
R4
O
R2
(c)
(d), (e)
H
Br
O
H
N
R4
R1
Br
S
O
O
8
9
O
R2
O
R2
R5
R2
S
OH
R5
O
R2
S
HO
O
HO
R3
N
B
R3
N
HO
R4
R4
(f)
R1
R1
Br
+
O
O
O
O
10
11
12-47
Scheme 1. Reagents and conditions: (a) K2CO3, DMF, microwave 170 °C, 10 min; (b) chlorosulfonic acid, 0 °C to rt, 4 h; (c) pyridine, 16 h; (d) R3Br, K2CO3, DMF, microwave
170 °C, 7 min; (e) LiOH, THF/H2O; (f) Pd(dppf)2Cl2, Cs2CO3, THF:H2O (1:1), microwave 160 °C, 10 min.
the resulting sulfonyl chloride 7 was combined with the appropri-
ate aniline 8 in pyridine to afford sulfonamide 9. Alkylation of the
sulfonamide nitrogen with an alkyl halide in DMF under micro-
wave conditions, followed by hydrolysis of the methyl ester affor-
ded the phenoxyacetic acid intermediate 10 which was then
coupled with a variety of boronic acids 11 in parallel to produce
the final products 12–47. This synthetic strategy proved to be a
highly efficient and robust method for rapid SAR identification
for this series, using one synthetic sequence to explore four points
of diversity simultaneously. All final compounds were purified by
reversed-phase HPLC to purities of >95% (LC–MS, UV 214 nm
detection). PPARd binding affinity and subtype selectivity were
then determined in an in vitro ligand displacement assay7 and
functional PPARd activity was evaluated in a standard Gal4 chi-
mera cell-based reporter assay.8 The results are summarized
below.
adjacent to the AF-2 helix where the phenoxyacetic acid head-
group binds and participates in a key hydrogen bonding network.9
Therefore, incorporating alkyl substituents adjacent to the carbox-
ylic acid to disrupt stabilization of the AF-2 helix could lead to par-
tial efficacy. A similar strategy was reported by researchers at Novo
Nordisk.10 We discovered that potency and efficacy could be mod-
ulated by changing the substituents at this position. The des-
methyl analogue 12 was the most potent, but displayed the highest
level of PPARd agonist activity (85%). Lengthening the R1 alkyl
group resulted in further erosion of binding affinity and functional
activity (13–15). Chiral separation of compound 3 generated com-
pounds (S)-3a and (R)-3b.11 Data from the cell-based reporter as-
say demonstrated a distinct difference in functional potency but
similar efficacy for these two compounds, with the R-enantiomer
(3b) displaying superior potency.
Efforts to crystallize PPARd in complex with partial agonists
such as 3 were unsuccessful. However, crystals were obtained with
more agonistic compounds such as 48 (hPPARd binding pIC50 = 7.5,
n = 3, hPPARd pEC50 = 8.1, 90%max).12
Initially the SAR at the alpha position of the phenoxyacetic acid
(R1) was investigated (Table 1). Ligand bound crystal structures
have shown that PPARd possesses a narrow lipophilic pocket

K. A. Evans et al. / Bioorg. Med. Chem. Lett. 21 (2011) 2345–2350
2347
Table 1
the compound or the AF2 helix to shift slightly in each case, possi-
bly attenuating the interactions that hold the AF2 helix in the ac-
tive position. This could in turn increase the mobility of the AF2
helix, while reducing the level of activation and the degree of agon-
ism. This mobility might also explain why we were unable to crys-
tallize other, less agonistic, partial agonists.
Investigations of substitution of the phenoxyacetic phenyl acid
ring at the R2 position revealed that replacement of the 2,3-
dimethyl substituents in compound 3 with a fused cyclohexyl ring
(16) provided an analogue which retained potency and partial effi-
cacy (hPPARd pIC50 = 7.8, pEC50 = 7.8, 62%). All other substitutions,
both electron-donating (18, 20, 22) and electron-withdrawing (17,
19, 21) resulted in less potent compounds (Table 2).
hPPAR binding and functional potency: alpha position (R1) variationsa
O
O
R1
Cl
H
O
R1
N
S
O
O
Compound
R1, R1
hPPAR binding pIC50
hPPARd reporter
a
c
d
pEC50
%Max
12
3
H, H
Me, H
Et, H
5.2b
4.7
4.8
5.0
5.0
4.8
4.7
5.6b
5.1
5.3
5.5
5.4
5.2
5.2
7.9
8.7
85
56
51
––
––
57
59
7.3
7.6b
6.9d
<5.0
<5.0
6.5
13
14
15
3a
3b
7.2c
6.8
Pr, H
Several alkyl variants at the sulfonamide nitrogen (R3) were
prepared to explore steric effects on potency and efficacy. Addition
as well as removal of a methylene unit from the linear butyl chain
led to a decrease in potency (23 and 24). Compounds with hetero-
atom substituted linear chains four atoms in length (27 and 28), as
well as small cycloalkyl-substitutions (25 and 26) were active, but
Bu, H
S-Me, H
R-Me, H
6.8c
6.2c
7.6c
7.9
a
Values are means of at least two experiments, standard deviation <0.2 unless
otherwise noted.
b
SD = 0.3.
SD = 0.4.
SD = 0.5.
c
d
Table 2
hPPAR binding and functional potency: R2 position variationsa
O
S
O
HO
O
2 R2
N
S
O
Cl
3
HO
O
O
N
S
48
O
O
Compound
R2
hPPAR binding pIC50
hPPARd reporter
a
c
d
pEC50
%Max
The crystal structure shows that 48 binds similarly to previous
agonists such as 1, and makes similar interactions with the key
acid-binding residues ( Fig. 2). The R- and S-methyl compounds
3a and 3b were overlaid into PPARd using the crystal structure of
48 as a template. In these overlays, the S-methyl group of 3a lies
2.1 Å from Thr289, whereas the R-methyl group of 3b lies 2.9 Å
from Leu469 in the AF2 helix.13 This repulsion is more severe in
3a than 3b, consistent with their potencies, and would force either
16
3
Cyclohexyl
2,3-diMe
2,3-diCl
2-OMe, 3-Cl
2-F, 3-Cl
2-F, 3-OMe
2,3-diF
2,3-diOMe
4.8
4.7
5.0
4.8
4.9
4.8
5.0
<4.6
5.1
5.1
5.4
5.2
5.2
5.3
5.5
5.4c
7.8c
7.3
7.1
6.8
6.6
6.3
6.2
5.3
7.8
62
56
70
70
81
53
73
7.6b
7.2
17
18
19
20
21
22
6.5
6.4b
6.3
6.5
<5.0
a
Values are means of at least two experiments, standard deviation <0.2 unless
otherwise noted.
b
SD = 0.3.
SD = 0.4.
c
Table 3
hPPAR binding and functional potency: R3 position variationsa
O
O
Cl
R3
N
H
O
S
O
O
Compound R3
hPPAR binding pIC50
hPPARd reporter
a
c
d
pEC50
%Max
3
n-Butyl
n-Propyl
n-Pentyl
Cyclobutylmethyl
Cyclopropylmethyl 4.8
2-OMe-ethyl
CF3propyl
4.7
4.7
4.8
4.9
5.1
5.1
5.3
5.2
5.1
4.9
5.3
7.3
6.9
6.1
6.0
6.1
6.3
5.9
7.6b
7.1b
6.4
56
55
50
59
53
48
56
23
24
25
26
27
28
6.3b
6.1
Figure 2. Crystal structure of 48 complexed with PPARd (green carbons) overlaid
onto structure of 1 (blue carbons) and the S- and R-methyl groups of 3a (pink) and
3b (purple), showing the AF2 helix (ribbon representation) and key side-chains
<4.7
4.7
6.3
5.7
(numbered by homology with PPARc). Hydrogen bonds with the acid headgroup
a
Values are means of at least two experiments, standard deviation <0.2 unless
shown with gray dots, and repulsive interactions with Thr289 and Leu469 shown
with black dots. Nitrogen, oxygen, sulfur and fluorine shown in blue, red, yellow
and gray, respectively.
otherwise noted.
b
SD = 0.3.

2348
K. A. Evans et al. / Bioorg. Med. Chem. Lett. 21 (2011) 2345–2350
none were as potent as the parent butyl group in compound 3
(Table 3).
30 retained excellent potency and partial efficacy comparable to
the original 2-methyl compound 3. The 2-chloro compound 29
was the most potent partial agonist analogue (hPPARd binding
pIC50 = 8.1, hPPARd pEC50 = 8.0, 53%max) in this set.
Substitution of the N-phenyl sulfonamide ring was also ex-
plored (Table 4). In general, substitutions made at the C4 (32 and
33) and/or C6 (36–38) position of the aryl ring led to decreased
binding affinity and reduction in functional efficacy with substitu-
tion at C6 (34 and 35) being more detrimental. These substitutions
may have steric effects on the adjacent para-chloro phenyl ring or
sulfonamide group that induce conformational changes that are
unfavorable to positive binding interactions. Substitution at C5
was acceptable as evidenced by the CF3 analogue 31 which main-
tained potent binding affinity. Substitution at C2 was well toler-
ated. For example, the 2-chloro and 2-methoxy analogues 29 and
Finally, R5 substitutions on the right-hand side phenyl ring were
investigated (Table 5). Substitution at the para-position of the aro-
matic ring was most preferred, with electron-withdrawing groups
exhibiting modest improvement in potency as compared to elec-
tron-donating groups (3, 42 vs 43–45). Chloro-substituted
analogues 3, 39, and 41 further illustrate substitution at the
para-position as optimal.
Using the aforementioned SAR data, compounds 46 and 47 were
subsequently designed, prepared and evaluated. The in vitro data
for these compounds is shown in Table 6. As expected, both com-
pounds showed increased hPPARd potency (pEC50 = 8.6), yet still
retained the desired partial agonist profile.
Table 4
hPPAR binding and functional potency: R4 position variationsa
Table 6
hPPAR binding and functional potency for compounds 46 and 47
O
O
O
Cl
HO
R5
O
2
HO
N
Cl
S
N
O
O
6
S
4
5 R4
O
O
R4
hPPAR binding pIC50
hPPARd reporter
Compound
R5
hPPAR binding pIC50
hPPARd reporter
a
Compound
a
c
d
pEC50
%Max
a
c
d
pEC50
%Max
3
2-Me
2-Cl
2-OMe
5-CF3
4-F
4-Me
6-OCF3
6-Me
4,6-diF
4-F, 6-Me
2,4,6-triMe
4.7
4.9
4.9
4.9
4.7
4.7b
4.9
5.0d
5.0
5.0
4.7b
5.1
5.3
5.1
5.3
5.7
5.5
5.0
5.2
5.8
5.1
5.1
7.3
7.6c
8.0
7.7
7.0
7.4
6.9
6.0
6.1
5.9
5.5
<5.0
56
53
66
69
48
41
30
36
27
25
46
47
OCF3
CF3
5.0
5.1
5.2
5.2
7.8
8.6
51
59
8.1e
7.6d
7.2
8.2c
8.6b
29
30
31
32
33
34
35
36
37
38
a
Values are means of at least six experiments, standard deviation <0.2 unless
otherwise noted.
6.7
b
6.4e
5.5d
5.8
SD = 0.3.
SD = 0.4.
c
5.5
5.3
5.0
CPT1a
a
80
70
60
50
40
30
20
10
0
Values are means of at least two experiments, standard deviation <0.2 unless
otherwise noted.
b
Values are n = 1.
SD = 0.3.
SD = 0.4.
SD = 0.5.
c
pEC50 = 8.4
d
e
compound 46
compound 47
pEC50 = 8.0
Table 5
hPPAR binding and functional potency: R5 position variationsa
0.0001 0.001
0.01
0.1
1
O
compound concentration (μM)
O
HO
R5
N
S
PDK4
100
90
80
70
60
50
40
30
20
10
0
O
O
Compound
R5
hPPAR binding pIC50
hPPARd reporter
a
c
d
pEC50
%Max
pEC50 = 8.0
pEC50 = 7.6
3
4-Cl
3-Cl
4.7
5.1
4.7
4.9
4.8
5.0
4.8
4.8
5.1
5.4
5.3
5.3
5.2
5.3
5.3
5.2
7.3
6.2
6.9
5.8
7.8
7.1
7.1
7.0
7.6c
6.3
56
42
47
34b
64
49
59
56
39
40
41
42
43
44
45
compound 46
compound 47
3,4-diCl
2-Cl
4-CF3
4-OCF3
4-Me
4-OMe
6.9
5.7b
8.2c
7.4d
7.5
0.0001
0.001
0.01
0.1
1
7.4
a
Values are means of at least two experiments, standard deviation <0.2 unless
compound concentration (uM)
otherwise noted.
b
Values are n = 1.
SD = 0.3.
SD = 0.4.
Figure 3. Expression of PPARd target genes for compounds 46 (pEC50 = 8.0, 7.6) and
47 (pEC50 = 8.4, 8.0) against CPT1a and PDK4 respectively. Each curve is expressed
as a percent of the full agonist GW610742X3 at 10 nM.
c
d

K. A. Evans et al. / Bioorg. Med. Chem. Lett. 21 (2011) 2345–2350
2349
Compounds 46 and 47 were studied for their effects on the
expression of the PPARd-regulated genes CPT1a and PDK4 in hu-
man skeletal muscle cells as previously described.14 The target
gene CPT1a is an important regulator of fatty acid b-oxidation in
skeletal muscle cells, and PDK4 plays a key role in skeletal muscle
metabolism by contributing to the regulation of glucose metabo-
lism. Both analogues 46 and 47 profiled as exceptionally potent
partial agonists, inducing endogenous PPARd target gene expres-
sion consistent with their profiles in the cell-based reporter assay
(Fig. 3).
Compounds 46 and 47 also had good physicochemical proper-
ties (aqueous solubility = 140 M, artificial membrane permeabil-
l
ity = 250 nm/s) and no significant activity against the common
cytochrome P450 (CYP450) isoforms (1A2, 2C19, 2D6, 3A4
pIC50 <5.5) except 2C9 (pIC50 = 6.1). In vitro studies using human li-
ver microsomes showed that both compounds had low clearance
(0.028 and 0.018 mL/min/mg, respectively). In oral mouse pharma-
cokinetic (PK) studies, both compounds showed good oral expo-
sure and half-life (Table 7). Collectively, this data provided an
excellent basis for further in vivo testing of these compounds for
efficacy.
Table 7
Compounds 46 and 47 were therefore chosen for in vivo evalu-
ation based on their partial induction of PPARd-responsive genes in
human muscle cell cultures and favorable PK profile.15 At the
100 mg/kg dose, both compounds decreased body weight gain
( Fig. 4), and at the 30 and 100 mg/kg dose, both compounds
showed a significant improvement in glucose sensitivity by OGTT
in a dose-dependent manner in the obese, insulin-resistant ob/ob
mouse (Fig. 5).16 These effects were similar to those induced by
In vitro and in vivo mouse DMPK results for compounds 46 and 47
Clint (mL/min/mg)a
AUC0–inf (ng h/mL)
T1/2 (h)
b
b
Compound
46
47
0.028
0.018
5870
2475
5.4
4.3
a
Intrinsic clearance in human liver microsomes.
In vivo mouse DMPK parameters: n = 2, two male CD rats dosed at 10 mg/kg po,
b
vehicle formulation 0.5%HPMC/0.1% Tween 80.
12
10
8
Vehicle
GW610742X(30 mg/kg)
47 (10 mg/kg)
47 (30 mg/kg)
47 (100 mg/kg)
46 (10 mg/kg)
46 (30 mg/kg)
46 (100 mg/kg)
6
4
2
0
0
2
5
7
9
11
13
15
Day
Figure 4. Effects of compounds 46 and 47 on body weight. Vehicle formulation 0.5% HPMC/0.1% Tween80 (n = 8/dose group).
350
300
Vehicle
250
GW610742X
(30 mg/kg)
47 (10 mg/kg)
47 (30 mg/kg)
47 (100 mg/kg)
46 (10 mg/kg)
46 (30 mg/kg)
46 (100 mg/kg)
200
150
100
50
0
0
15
30
60
90
-50
Timepoint (min)
Figure 5. Effects of compounds 46 and 47 in oral glucose tolerance test (OGTT). Vehicle formulation 0.5% HPMC/0.1% Tween80 (n = 8/dose group).

2350
K. A. Evans et al. / Bioorg. Med. Chem. Lett. 21 (2011) 2345–2350
Merrihew, R. V.; Huet, S.; Willson, T. M. Bioorg. Med. Chem. Lett. 2008, 18, 5018;
(b) Sauerberg, P.; Olsen, G. S.; Jeppesen, L.; Mogensen, J. P.; Pettersson, I.;
Jeppesen, C. B.; Daugaard, J. R.; Galsgaard, E. D.; Ynddal, L.; Fleckner, J.;
Panajotova, V.; Polivka, Z.; Pihera, P.; Havranek, M.; Wulff, E. M. J. Med. Chem.
2007, 50, 1495.
the full agonist GW610742X. These data suggest that PPARd partial
agonists such as 46 and 47 (pEC50 = 8.6, 51% and 59%max, respec-
tively) can deliver anti-diabetic efficacy in mice.
In summary, a series of phenoxyacetic acids as subtype selective
and potent hPPARd partial agonists has been developed. Systematic
structural modifications of this template at five positions of diver-
sity were achieved through one primary solution-phase synthesis
which resulted in the rapid identification of key structure–activity
relationships (SAR), and the discovery of two potent exemplars.
Compounds 46 and 47 were further evaluated in vivo, and at both
the 30 and 100 mg/kg dose, both compounds decreased body
weight gain and significantly improved insulin sensitivity by OGTT
in the obese, insulin-resistant ob/ob mouse suggesting that PPARd
partial agonists can deliver anti-diabetic efficacy in mice.
5. Examples of the phenoxyacetic acid template have also been established by
others as full agonist ligands for PPARd, for example: (a) Luckhurst, C. A.;
Ratcliffe, M.; Stein, L.; Furber, M.; Botterell, S.; Laughton, D.; Tomlinson, W.;
Weaver, R.; Chohan, K.; Walding, A. Bioorg. Med. Chem. Lett. 2011, 21, 531; (b)
Luckhurst, C. A.; Stein, L. A.; Furber, M.; Webb, N.; Ratcliffe, M. J.; Allenby, G.;
Botterell, S.; Tomlinson, W.; Martin, B.; Walding, A. Bioorg. Med. Chem. Lett.
2011, 21, 492; (c) Connors, R. V.; Wang, Z.; Harrison, M.; Zhang, A.; Wanska, M.;
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Acknowledgments
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The authors thank Walter Johnson and Chad Quinn for their
assistance with the LC–MS spectra, Ann McFarland and Doug
Minick for their assistance with chiral separation and analysis,
Jane Blankenhagen for her assistance with in vitro DMPK, and
Lisa M. Shewchuk for additional refinement of the crystal
structure.
Supplementary data
10. Pettersson, I.; Ebdrup, S.; Havranek, M.; Pihera, P.; Korinek, M.; Mogensen, J. P.;
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11. The enantiomers were purified and separated by SFC using 10% MeOH/0.1%
TFA, 90% CO2, 140 bar, 40 °C, 2 mL/min on an AS column monitored at 215 nm.
(S)-3a, 95% ee, 87% pure and (R)-3b, 98% ee, 93% pure.
Supplementary data (synthetic procedures and characterization
data for compounds 46 and 47) associated with this article can be
found, in the online version, at doi:10.1016/j.bmcl.2011.02.077.
References and notes
12. The crystal structure of 48 was solved at 2.4 Å resolution, R = 20.0%,
Rfree = 23.6%, PDB code 3PEQ.
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13. Distances reported are the average distances from the residues in the crystal
structure of 48 bound to PPARd which has two subunits. Compounds 3a and 3b
were overlaid into both subunits, with the (S)-methyl of 3a lying 2.01 and
2.26 Å from Thr289 in the two subunits, and with the (R)-methyl of 3b lying
2.78 and 2.93 Å from Leu469 in the two subunits.
14. Shearer, B. G.; Steger, D. J.; Way, J. M.; Stanley, T. B.; Lobe, D. C.; Grillot, D. A.;
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15. All animal studies were conducted after review by the GSK Institutional Animal
Care and Use Committee and in accordance with the GSK Policy on the Care,
Welfare and Treatment of Laboratory Animals.
16. ob/ob mice, 6 weeks of age. Positive control: GW610742X (30 mg/kg). Vehicle:
HPMC/Tween. Compound dosed orally (10, 30, 100 mg/kg) once daily for
15 days. OGTT performed on fasted animals at day 12 of dosing.