Design and synthesis of N-[(4-methoxyphenoxy)carbonyl]-N-[[4-[2-(5-methyl- 2-phenyl-4-oxazolyl)ethoxy]phenyl]methyl]glycine [muraglitazar/BMS-298585], a novel peroxisome proliferator-activated receptor α/γ dual agonist with efficacious glucose and lipid-l

Article abstract of DOI:10.1021/jm0496436

Muraglitazar/BMS-298585 (2) has been identified as a non-thiazolidinedione PPAR α/γ dual agonist that shows potent activity in vitro at human PPARα (EC₅₀ = 320 nM) and PPARγ (EC₅₀ = 110 nM). Compound 2 shows excellent efficacy for lo

Full text of DOI:10.1021/jm0496436

2248
J. Med. Chem. 2005, 48, 2248-2250
Design and Synthesis of N-[(4-Methoxyphenoxy)carbonyl]-N-[[4-[2-(5-
methyl-2-phenyl-4-oxazolyl)ethoxy]phenyl]methyl]glycine [Muraglitazar/
BMS-298585], a Novel Peroxisome Proliferator-Activated Receptor r/γ Dual
Agonist with Efficacious Glucose and Lipid-Lowering Activities
Pratik V. Devasthale,*,† Sean Chen,† Yoon Jeon,† Fucheng Qu,† Chunning Shao,† Wei Wang,† Hao Zhang,†
Michael Cap,‡ Dennis Farrelly,‡ Rajasree Golla,‡ Gary Grover,‡ Thomas Harrity,‡ Zhengping Ma,‡ Lisa Moore,‡
Jimmy Ren,‡ Ramakrishna Seethala,‡ Lin Cheng,‡ Paul Sleph,‡ Wei Sun,‡ Aaron Tieman,‡ John R. Wetterau,‡
Arthur Doweyko,§ Gamini Chandrasena,# Shu Y. Chang,# W. Griffith Humphreys,# Vito G. Sasseville,|
Scott A. Biller,† Denis E. Ryono,† Fred Selan,| Narayanan Hariharan,‡ and Peter T. W. Cheng*,†
Metabolic Diseases Chemistry, Metabolic Diseases Biology, Macromolecular Structure, Metabolism and Pharmacokinetics,
Drug Safety Evaluation, Bristol-Myers Squibb Pharmaceutical Research Institute, Princeton, New Jersey 08543-5400
Received May 13, 2004
Muraglitazar/BMS-298585 (2) has been identified as a non-thiazolidinedione PPAR R/γ dual
agonist that shows potent activity in vitro at human PPARR (EC50 ) 320 nM) and PPARγ
(EC50 ) 110 nM). Compound 2 shows excellent efficacy for lowering glucose, insulin,
triglycerides, and free fatty acids in genetically obese, severely diabetic db/db mice and has a
favorable ADME profile. Compound 2 is currently in clinical development for the treatment of
type 2 diabetes and dyslipidemia.
Introduction
Type 2 diabetes mellitus is a chronic and devastating
disease characterized by hyperglycemia, insulin resis-
tance, and perturbations in fat, protein, and carbohy-
drate metabolism. According to the World Health Or-
ganization, approximately 150 million people are afflicted
with the disease worldwide with projections of 300
million by the year 2025.1 Risk factors for type 2
diabetes include obesity, genetic predisposition, physical
inactivity, history of gestational diabetes, impaired
glucose tolerance, race, and ethnicity. Patients with type
2 diabetes often also suffer from dyslipidemia in the
form of high plasma triglycerides and low HDL-
cholesterol levels, both considered risk factors for coro-
nary heart diseases.2 Current treatments for type 2
diabetes include biguanides, sulfonylureas, insulin for-
mulations, R-glucosidase inhibitors, insulin sensitizers,
and insulin secretagogues.3 Two thiazolidinediones,
Figure 1. Design of oxybenzylglycines.
rosiglitazone4 and pioglitazone,5 representing a novel
class of insulin-sensitizing peroxisome proliferator-
activated receptor γ (PPARγ) agonists have been in
clinical use as antidiabetic drugs. Fibrates (e.g., fenofi-
brate and gemfibrozil), which lower triglycerides and
elevate HDL levels, are weak PPARR agonists and have
been in clinical use for the treatment of dyslipidemia.
A dual PPARR/γ agonist that improves insulin sensitiv-
ity, lowers glucose, and corrects lipoprotein abnormali-
ties would be of great interest as a drug for the
treatment of type 2 diabetes. A number of PPAR
agonists (R- and γ-selective; R/γ dual) have been or
currently are in clinical development.6-9 AZ-2426 and
KRP-297/MK-7677 are PPARR/γ dual agonists, while GI-
262570/farglitazar9 is a selective PPARγ agonist and
GW-957810 is a PPARR-selective agonist. In this paper
we describe the design, synthesis, and in vivo charac-
terization of 2, a novel, non-thiazolidinedione PPARR/γ
dual agonist that shows promise for the treatment of
type 2 diabetes and the associated dyslipidemia.
Our initial lead, the oxybenzylglycine 1, was concep-
tually derived via homologation of a novel azaisostere
structure drawn from known R-alkoxy and R-aminoaryl-
propanoic acid PPAR ligands (Figure 1).6,9,11,12 Signifi-
cant advantages gained from this change include the
elimination of a chiral center, simplification of synthesis,
and opportunities for rapid generation of diversity from
the lead structure. Thus, exploration of the replacement
of the N-benzyl moiety of 1 was carried out in an effort
to optimize the relatively weak in vitro potency of 1. One
of the approaches involved the synthesis of a set of
carbamate acids, facilitated by iterative solution-phase
parallel synthesis. Several of these carbamate acids
showed promising oral in vivo activity, and 2 was
identified from this effort as an optimized candidate for
* To whom correspondence should be addressed. For P.V.D.:phone,
609-818-4974; fax, 609-818-3460; e-mail, pratik.devasthale@bms.com.
For P.T.W.C.: phone, 609-818-4952; fax, 609-818-3460; e-mail,
peter.t.cheng@bms.com.
† Metabolic Diseases Chemistry.
‡ Metabolic Diseases Biology.
§ Macromolecular Structure.
# Metabolism and Pharmacokinetics.
| Drug Safety Evaluation.
10.1021/jm0496436 CCC: $30.25 © 2005 American Chemical Society
Published on Web 11/13/2004

Brief Articles
Journal of Medicinal Chemistry, 2005, Vol. 48, No. 6 2249
Scheme 1. Synthesis of 2a
Table 2. In Vitro Transactivation Activity of 2 in 3T3L-1
Preadipocyte Cells (PPARγ)
triglycerides in
compds
cell lysate (mg/dL)a
vehicle
15 ( 1
46 ( 2
13 ( 2
48 ( 3
2 (R/γ)
WY-14643 (R)
rosiglitazone (γ)
a 3T3L-1 cells were incubated in the presence of 1.0 µM test
compound.
Table 3. Glycemic and Lipid Lowering Effects of 2 in Male
db/db Mice
2
vehicle
(n ) 10 mice/group)
(10 mg kg-1 day-1
14 days)
,
plasma parameters
glucose (mg/dL)
259.0 ( 24.0
124.0 ( 5.0
0.67 ( 0.07
3.9 ( 0.6
120.0 ( 8.0 (-54%a)
84.0 ( 7.0 (-33%a)
0.25 ( 0.06 (-62%a)
2.0 ( 0.2 (-48%a)
triglycerides (mg/dL)
free fatty acids (mequiv/L)
insulin (ng/mL)
a Difference between vehicle and drug treated groups; p e 0.05.
Table 4. Pharmacokinetic Profile of 2 in Male SD Ratsa
oral
10.0
intra-arterial
a (a) MeSO2Cl, Et3N, CH2Cl2, 25 °C, 1 h, >98%; (b) 4-hydroxy-
benzaldehyde, K2CO3, CH3CN, 95 °C, 24 h, 71%; (c) H2NCH2-
CO2Me‚HCl, Et3N, 4 Å molecular sieves, anhydrous MgSO4,
MeOH, 25 °C, 12 h; NaBH4, 25 °C, 3.5 h, 80%; (d) 4-methoxyphenyl
chloroformate, Et3N, CH2Cl2, 25 °C, 2 h, 98%; (e) LiOH‚H2O, THF/
H2O (1:1), 25 °C, 2.5 h, 94%.
dose (mg kg-1
)
5.0
27.7 ( 3.8
AUC∞ (µg h mL-1
)
49.0 ( 7.3
20.8 ( 2.4
0.4 ( 0.1
Cmax (µg mL-1
Tmax (h)
)
T1/2 (h)
7.3 ( 4.0
3.0 ( 0.4
0.6 ( 0.2
Cl (mL min-1 kg-1
)
Vss (L kg-1
F (%)
)
Table 1. In Vitro Receptor Binding and Transactivation
Activity Profile of 2a
88
a n ) 3.
hPPARRb
hPPARγb
IC50
EC50
IC50
EC50
considerably more potent than the fibrates (e.g., fenofibric
acid) at PPARR. There was no cross-reactivity against
other nuclear hormone receptors such as PPARδ, RXRR,
RARs, ERR/â, AR, and PR. Furthermore, in a pre-
adipocyte differentiation assay (Table 2), which mea-
sures the extent of predominantly PPARγ-mediated
differentiation of preadipocytes into triglyceride-loaded
adipocytes, 2 induced a 3-fold increase in differentiation,
indicating potent activation of PPARγ. This level of
activity was comparable to that of rosiglitazone, a
known selective PPARγ agonist. WY-14643, a potent
and selective PPARR agonist, was used as a negative
control in this assay.
1
2
1.4
0.25 ( 0.08
0.02 (103%) 4.8
1.5 (111%)
0.11 ( 0.06
(82 ( 4%)b
0.14 (100%)
0.32 ( 0.1
(70 ( 4%)b
>32 µM
0.19 ( 0.07
rosiglita- >100 µM
0.25
zone
GW-2331 0.538 ( 0.165 <0.02
0.316 ( 0.126 0.425 ( 0.07
(64 ( 3%)
(102 ( 3%)
>10.0 (30%) >25 µM
fenofibric >10.0
>100 µM
acid
a IC50 and EC50 are in µM and are defined in the Experimental
Section. b Intrinsic activity at 1 µM relative to a primary standard
(at 1 µM), expressed as a percentage, is represented in parenthe-
ses. GW-233114 and rosiglitazone15 were used as primary stan-
dards for PPARR and PPARγ activity, respectively.
An in vivo study of 2 was carried out in genetically
obese male db/db mice (at a dose of 10 mpk/day for 14
days). Compound 2 was highly efficacious in this study,
reducing levels of glucose (-54%, representing normal-
ization), triglycerides (-33%, representing normaliza-
tion), nonesterified fatty acids (-62%), and insulin
(-48%) (Table 3). Glucose normalization with concomi-
tant reduction in insulin levels suggests insulin sensi-
tizing antidiabetic effects through PPARγ activation.
Presumably, the triglyceride lowering effect is predomi-
nantly mediated through PPARR activation.
Compound 2 has an excellent ADME profile that is
suitable for clinical development (Table 4). It has very
good oral bioavailability in rats (88%), with a reasonable
plasma t1/2 of 7.3 h and low systemic clearance of 3 mL
min-1 kg-1. In male beagle dogs and cynomolgus
monkeys, the oral bioavailability was 18% and 79%,
respectively. The oral absorption was rapid, with Tmax
occurring at 0.6 h in both species. The corresponding
Cmax values were 0.12 µg/mL in dogs (1 mg/kg oral dose)
and 15.7 µg/mL in monkeys (2 mg/kg oral dose).
In conclusion, 2 is a potent, novel non-thiazolidinedi-
one PPARR/γ dual agonist in vitro that demonstrates
in vivo efficacy and ADME profiling. Scheme 1 describes
a scalable preparation of 2.
Chemistry
Alkylation of 4-hydroxybenzaldehyde with the phen-
yloxazole mesylate 4, prepared readily from commer-
cially available alcohol 3, yielded aldehyde 5, which was
reacted with glycine methyl ester under standard
reductive amination conditions13 to provide secondary
amine 6 in excellent yield. Reaction of amine 6 with
4-methoxyphenyl chloroformate followed by hydrolysis
of the methyl ester afforded 2 in 94% yield.
Results and Discussion
As shown in Table 1, 2 binds with high affinity to
PPARR (IC50 ) 250 nM) and PPARγ (IC50 ) 190 nM).
In transactivation assays 2 shows potent and novel
functional activity at the full-length human receptors
for PPARR (EC50 ) 320 nM; intrinsic activity at 1 µM
is 70%) and PPARγ (EC50 ) 110 nM; intrinsic activity
at 1 µM is 82%). Thus, 2 shows functional activity
comparable to that of rosiglitazone at PPARγ and is

2250 Journal of Medicinal Chemistry, 2005, Vol. 48, No. 6
Brief Articles
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treatment of diabetes and associated dyslipidemia.
PPARR/γ dual agonists such as 2 may also be of utility
in establishing a therapeutic modality for the treatment
of metabolic syndrome (which is characterized by im-
paired glucose tolerance, hyperinsulinemia, dyslipi-
demia, and hypertension).
2
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[(4-Methoxyphenoxycarbonyl)-{4-[2-(5-methyl-2-phen-
yloxazol-4-yl)ethoxy]benzyl}amino]acetic Acid (2). To a
10 °C solution of amine 6 (16.6 g, 43.7 mmol) in 200 mL of
CH2Cl2 were successively added Et3N (9.13 mL, 65.5 mmol)
and 4-methoxyphenyl chloroformate (7.65 mL, 50.2 mmol)
dropwise while maintaining the reaction temperature at e12
°C. The mixture was allowed to warm to 25 °C and stirred for
2 h. Volatiles were removed in vacuo, and the residue was
chromatographed (SiO2; 4:1 to 1:1 hexanes/EtOAc) to afford 2
methyl ester (22.8 g, 98%) as a colorless, viscous oil.
To a solution of the above methyl ester (18.75 g, 35.4 mmol)
in THF (275 mL) was added a solution of LiOH‚H2O (4.44 g,
105.9 mmol) in 275 mL of H2O. The resulting mixture was
stirred at 25 °C for 2.5 h, after which the pH was adjusted to
∼4 with 1 N aqueous HCl. The THF was removed in vacuo,
the residue was diluted with 800 mL of EtOAc, and the
mixture was stirred for 0.5 h at 25 °C. The organic phase was
separated, dried (MgSO4), and concentrated in vacuo to afford
a solid, which was recrystallized from EtOH to give 2 (8.41 g)
as a colorless solid. An additional 8.76 g of product was
recovered in two subsequent recrystallizations from the mother
liquor to give 2 (17.2 g total) in 94% yield. Mp: 139.1-140.2
°C. 1H NMR (400 MHz, CDCl3): rotamers, δ 2.38/2.39 (s, 3H,
oxazole-CH3), 3.00/3.01 (s, 2H, oxazole-CH2), 3.77/3.78 (s, 3H,
ArOCH3), 4.02 (s, 2H, -CH2COOH), 4.20-4.24 (m, 2H,
-CH2CH2OAr), 4.55 (s, 1H, ArCHaHbN), 4.65 (s, 1H, Ar-
CHaHbN), 6.8-7.0 (m, 4H), 7.0-7.1 (m, 2H), 7.2-7.3 (m, 2H),
7.40-7.45 (m, 3H), 7.96-7.98 (m, 2H). 13C NMR (100 MHz,
CDCl3): rotamers, δ 10.2 (oxazole-CH3), 26.0 (oxazole-CH2-),
47.3/47.5 (CH2COOH), 51.0/51.2 (ArCH2N), 55.6 (ArOCH3),
66.7 (oxazole-CH2CH2OAr), 114.3, 114.8, 122.5, 126.0, 127.3,
128.4, 128.7, 129.2, 129.9/130.1, 132.4, 144.8, 145.3, 155.4/
155.5, 157.1, 158.5, 159.7, 173.0. Anal. Calcd for C29H28N2O7:
C, 67.43; H, 5.46; N, 5.42. Found: C, 67.45; H, 5.47; N, 5.29.
IR (KBr): 2800-3200 (w, br), 1724 (vs), 1511 (s), 1197 (s), 1171
(s) cm-1. UV (MeOH, 13.9 mg/L): λmax 224, 277, 282 (sh), 290
(sh), 302 (sh) nm. LRMS (M + H+): 517.19. HRMS Calcd for
C29H29N2O7: 517.1975. Found: 517.1964.
Acknowledgment. The authors thank Dr. Carl
Decicco and Dr. Robert Zahler for proof-reading the
manuscript.
Supporting Information Available: Descriptions of
PPARR and -γ binding and transactivation assays, preadipo-
cyte differentiation assay, in vivo pharmacology; detailed
experimental procedures, physical data, elemental analyses,
IR, UV, 1H, 13C NMR, LRMS, and HRMS data for 5 and 6;
and synthesis procedure, 1H NMR, and LRMS data for 4. This
material is available free of charge via the Internet at http://
pubs.acs.org.
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Note Added after ASAP Publication. This manu-
script was released ASAP on 11/13/2004 with an incom-
plete author byline, with errors in the EC50 values in
the abstract, and with a labeling error in Scheme 1. The
correct version was posted on 12/6/2004.
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JM0496436