Design and synthesis of dual modulators of soluble epoxide hydrolase and peroxisome proliferator-activated receptors

Article abstract of DOI:10.1021/jm301194c

Metabolic syndrome is a complex condition which often requires the use of multiple medications as a treatment. The resulting problems of polypharmacy are increase in side effects, drug-drug interactions, and its high economic cost. Development of multitarget compounds is a promising strategy to avoid the complications arising from administration of multiple drugs. Modulators of peroxisome proliferator-activated receptors (PPARs) are established agents in the treatment of dyslipidaemia, hyperglycaemia, and insulin resistance. Inhibitors of soluble epoxide hydrolase (sEH) are under evaluation for their use in cardiovascular diseases. In the present study, a series of dual sEH/PPAR modulators containing a pyrrole acidic headgroup and a urea pharmacophore were designed, synthesized, and evaluated in vitro using recombinant enzyme and cell-based assays. Compounds with different activity profiles were obtained which could be used in the treatment of metabolic syndrome.

Full text of DOI:10.1021/jm301194c

Brief Article
pubs.acs.org/jmc
Design and Synthesis of Dual Modulators of Soluble Epoxide
Hydrolase and Peroxisome Proliferator-Activated Receptors
Estel.la Buscato,† Rene Blocher,† Christina Lamers, Franca-Maria Klingler, Steffen Hahn,
́
́
̈
Dieter Steinhilber, Manfred Schubert-Zsilavecz, and Ewgenij Proschak*
Institute of Pharmaceutical Chemistry, ZAFES/LiFF/OSF Goethe-University Frankfurt, Max-von-Laue-Strasse 9, D-60438 Frankfurt
am Main, Germany
S
* Supporting Information
ABSTRACT: Metabolic syndrome is a complex condition which often requires the use of multiple medications as a treatment.
The resulting problems of polypharmacy are increase in side effects, drug−drug interactions, and its high economic cost.
Development of multitarget compounds is a promising strategy to avoid the complications arising from administration of
multiple drugs. Modulators of peroxisome proliferator-activated receptors (PPARs) are established agents in the treatment of
dyslipidaemia, hyperglycaemia, and insulin resistance. Inhibitors of soluble epoxide hydrolase (sEH) are under evaluation for
their use in cardiovascular diseases. In the present study, a series of dual sEH/PPAR modulators containing a pyrrole acidic
headgroup and a urea pharmacophore were designed, synthesized, and evaluated in vitro using recombinant enzyme and cell-
based assays. Compounds with different activity profiles were obtained which could be used in the treatment of metabolic
syndrome.
linked to type 2 diabetes mellitus. The remaining subtype
PPARδ stimulates fatty acid oxidation in heart and skeletal
muscle and plays a role in cell differentiation and athero-
sclerosis. Reviewing recent patent literature implied that the
development of PPARδ agonists is a promising target toward
MetS associated pathologies.5
INTRODUCTION
■
The metabolic syndrome (MetS)1 is a clustering of factors
mainly consisting of the so-called “deadly quartet’’ of
hyperglycaemia, hypertriglyceridemia, hypertension, and obe-
sity. MetS results in an increased risk for atherosclerosis and
diabetes. Because of its complex nature, the current therapy
strategies of MetS require multiple treatments regulating lipid
and glucose homeostasis as well as blood pressure and
coagulation. Up-to-date treatment for MetS follows a lifestyle
change of the patient to increase the physical activity, initiation
of drug therapy with statins to reduce LDL, blood pressure
reducing agents, oral antidiabetics, and compounds to handle
obesity. Nevertheless, the combination of several drugs for
individual risk factors can decrease the efficacy and enhance the
toxicity of each drug. Thus, there is a strong unmet medical
need in reliable and efficient drugs targeting multiple symptoms
of MetS over the long term, thereby minimizing problems with
polypharmacy.2
Soluble epoxide hydrolase (sEH) metabolizes epoxyeicosa-
trienoic acids (EETs), previously produced by epoxygenases, to
the corresponding dihydroxyeicosatrienoic acids (DHETs).
Inhibition or deletion of soluble epoxide hydrolase prevents
hyperglycaemia, promotes insulin secretion, reduces islet
apoptosis, and decreases adipogenesis.6,7 On the basis of this
knowledge, sEH has attired the attention as possible target to
treat MetS8 in conjunction with selective PPAR activation. This
current study presents dual modulators that target sEH and
PPAR, an interesting therapeutic approach to treat MetS
related pathologies. PPARs and sEH are related through the
modulatory action of sEH substrates and metabolites on PPAR.
Latest investigations demonstrated that EETs9 are PPARγ
and PPARα ligands. Competition as well as direct binding
assays revealed that 14,15-EET10 binds to the ligand-binding
domain of PPARα.
Furthermore, latest publications discovered that DHETs
have a high potency to activate PPARα,10 suggesting that
DHETs may have additional vascular actions as a result of their
effect on PPARα. Figure 1 summarizes the expected mode of
action of dual acting compounds which should lead to
increased EETs levels due to sEH inhibition and substitute
the beneficial activation of PPAR by DHETs. Fang et al.
discovered an activation of PPARα by substituted urea-derived
sEHIs after cellular metabolism.11
PPARs contribute to the regulation of glucose, lipid, and
cholesterol metabolism, therefore they seem to be a valuable
target to treat MetS.3 With the hypolipidemic fibrates and the
antidiabetic thiazolidindiones (TZD), two drug classes had
entered the market. PPARγ agonists like troglitazone,
rosiglitazone, and most recently pioglitazone were suspended
by some authorities due to severe adverse events. However, a
very promising finding had been made when Choi et al.4
showed that the antidiabetic effect of the TZDs is partially
mediated by the inhibition of the cyclin-dependent kinase 5
mediated phosphorylation of PPARγ. With this discovery, a
promising new development regarding PPARγ as a drug target
arises.
The fibrate-derived PPARα agonists (e.g., clofibrate,
bezafibrate) are used in the treatment of dyslipidaemia
associated with atherosclerosis and dyslipidaemia primarily
Received: August 28, 2012
Published: November 7, 2012
© 2012 American Chemical Society
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Journal of Medicinal Chemistry
Brief Article
(5a−i) as carboxylic acid, they were solved in a mixture of
THF/MeOH/water (1:1:2) and treated with KOH under
microwave radiation to yield (6a−i).
RESULTS AND DISCUSSION
■
Known PPARα, γ, and δ selective agonists (GW7647,17
pioglitazone,18 L-16504119) and the sEH inhibitor AUDA13
were evaluated and used as references. To obtain dual
modulators of sEH/PPAR, it was necessary to link both
pharmacophores via an aromatic spacer and maintain the
lipophilic core which is characteristic of PPAR agonists.
Adamantyl and cyclohexyl ureas have been shown to exhibit
high sEH inhibitory properties. In terms of sEH inhibition, the
ethyl ester derivatives with cyclohexyl moieties (5a−c) are
better tolerated than adamantyl (5d−f) or Ph-pOCF3 (5g−i),
reaching IC50 values in the range from 23 to 39 nM. In general,
the introduction of a Ph-pOCF3 substituent decreased the
inhibitory activity, yielding inactive (5h, 6g) or slightly active
compounds (5g, 5i, 6h, 6i) (Table 1). The comparison of
carboxylic acids with the corresponding esters in terms of sEH
inhibition shows that cyclohexyl substituted esters (5a−c) and
adamantyl (5d−f) are more potent than their carboxylic acids
counterparts (6a−c, 6d−f). Regarding the substitution pattern,
the inhibitory potency of cyclohexyl carrying esters remains
unaffected (5a−c, 23 nM < IC50 < 39 nM). This tendency
holds true for adamantyl substituted esters (5d−f, 43 nM <
IC50 < 87 nM). This behavior stands in contrast with the
inhibitory potency of the acids, possibly due to an alternative
binding mode which already has been described for sEH.19
Regarding cyclohexyl acid derivatives o- (6a, IC50 = 747 nM)
and p- (6c, IC50 = 252 nM), substituted compounds were more
potent than the m-substituted moiety (6b, IC50 = 1923 nM). In
the case of adamantyl, the p- (6f, IC50 = 73 nM) was more
potent than o-adamantyl (6d, IC50 = 178 nM) and m- (6e, IC50
= 304 nM) derivatives, possibly due to sterical hindrance.
The PPAR activation potency of the compounds showed a
wide diversity from modulatory effects at 10 μM to full
agonistic properties with certain EC50 values (Table 2).
Regarding the PPAR activities, only two ethyl ester derivatives
showed a partial activation of PPARγ at 10 μM (5b, 21%; 5c,
16%). As expected, the majority of carboxylic acids (6a−i) were
able to activate PPARs. We observed that adamantyl derivatives
(6d−f) were able to activate PPARγ selectively, probably due to
the larger left distal ligand binding pocket compared to the
other receptor subtypes.16 The selectivity was impaired when
testing Ph-pOCF3 derivatives that activated both PPARα and
PPARγ concerning o-substitution (6g, 38% (PPARα) and
128%, EC50 = 2 μM (PPARγ)). While m- and p-substituted
compounds (6h, 6i) yield a PPARα full agonism, only partial
agonism was observed for the o-substituted Ph-pOCF3
Figure 1. Mode of action of dual sEH/PPAR modulators. The sEH
inhibitor leads to a decreased DHETs level, which might lead to
decreased PPAR activation. This effect is compensated by the sEH/
PPAR dual modulation.
In the present work, we describe novel compounds that are
able to inhibit sEH and activate PPAR regardless their
metabolism. Further development of these compounds could
lead to agents with beneficial action on hyperglycaemia,
hypertriglyceridemia, and hypertension, which can serve as a
starting point for the development of polypharmacological
compounds as a treatment of MetS.12
CHEMISTRY
■
We prepared the compounds 5a−i, 6a−i that are connected
through a methylbenzyl linker between the pyrrole ring and the
urea pharmacophore. An adamantyl group has been used in
many sEH inhibitors13 with high in vitro efficacy (e.g., AUDA),
and Ph-pOCF3 substituted moiety showed improved pharma-
cokinetic properties.14 On the basis of this knowledge, we
developed derivatives carrying pyrrole structures linked to urea
containing compounds (sEH pharmacophore) that end in a
carboxylic group needed for activation of all PPAR subtypes.
We followed the previous strategy15 to synthetize potent PPAR
agonists: acidic headgroup−aromatic core−linker−hydropho-
bic tail.16 In the present study, we explored the effect of the
substitution pattern (o-, m-, p-) on the activity at the different
targets. We synthesized the compounds 6a−i, with an acidic
group and 5a−i as ethyl carboxylate esters, as seen in Scheme 1.
Starting from ethyl 1H-pyrrole-2-carboxylate (1) that reacted
with the o-, m-, or p-(bromomethyl) benzonitrile (2) via phase
transfer catalysis using TBAI in a mixture of DCM/aqueous
NaOH (1/1), we obtained the following N-alkylated products
(3a−c) that underwent reduction with Raney Ni, the
corresponding amines (4a−c) remaining which reacted with
three different isocyanates: cyclohexylisocyanate, adamantyliso-
cyanate, and p-OCF3-phenylisocyanate, yielding the corre-
sponding urea derivatives (5a−i). To obtain the compounds
a
Scheme 1. Synthesis of sEH/PPAR Dual Modulators 5a−i, 6a−i
a
Reagents and conditions: (a) tetrabutylammonium iodide, NaOH 50% aqueous, DCM, 0 °C to rt, 12 h; (b) Raney Ni, H2, MeOH; (c) R-NCO,
DIPEA, DCM, rt, 12 h; (d) THF/MeOH/H2O (1:2:2), KOH, MW, 90 °C, 15 min.
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Journal of Medicinal Chemistry
Brief Article
a
Table 1. Inhibition and Activation Values of dual sEH/PPAR Modulators 5a−i, 6a−i
name
R1
R2
subst IC50 (nM) sEH % activation @10 μM PPARα % activation @10 μM PPARγ % activation @10 μM PPARδ
GW7647
nt
nt
100
nt
nt
pioglitazone
nt
100
nt
L-165041
AUDA
5a
nt
nt
nt
100
107 12.8
39 1.4
27 8.0
23 1.0
43 10.0
46 4.2
87 8.4
2009 1165.6
ia
nt
nt
nt
Et
Et
Et
Et
Et
Et
Et
Et
Et
H
H
H
H
H
H
H
H
H
cyclohexyl
cyclohexyl
cyclohexyl
adamantyl
adamantyl
adamantyl
Ph-pOCF3
Ph-pOCF3
Ph-pOCF3
cyclohexyl
cyclohexyl
cyclohexyl
adamantyl
adamantyl
adamantyl
Ph-pOCF3
Ph-pOCF3
Ph-pOCF3
o-
ia
ia
ia
5b
m-
p-
o-
ia
21 4.1
ia
5c
ia
16 4.6
ia
5d
ia
ia
ia
5e
m-
p-
o-
ia
ia
ia
5f
ia
ia
ia
5g
ia
ia
ia
5h
m-
p-
o-
ia
ia
ia
5i
611 248.4
747 194.9
1923 724.7
252 17.1
178 55.4
304 78.3
73 4.5
ia
ia
ia
ia
6a
ia
ia
ia
6b
m-
p-
o-
ia
ia
ia
6c
ia
ia
ia
ia
6d
45 14.7
16 5.7
18 7.8
105 14.4
43 12.5
84 24.1
ia
6e
m-
p-
o-
ia
ia
6f
ia
ia
6g
38 1.9
58 5.7
67 13.4
ia
25 4.5
ia
6h
m-
p-
943 658.3
258 48.0
6i
a
nt = not tested, ia = inactive.
a
Table 2. EC50 Values of Dual sEH/PPAR Modulators
name
IC50 (nM) sEH
EC50 (max activation) PPARα
EC50 (max activation) PPARγ
EC50 (max activation) PPARδ
GW7647
pioglitazone
L-165041
AUDA
6d
nt
nt
0.2 0.05 μM
nt
nt
nt
nt
nt
ia
0.2 0.05 μM
nt
nt
nt
nt
0.039 0.008 μM
107 12.8
178 55.4
ia
nt
ia
6
0.8 μM (60 1.3%)
6g
7
1.5 μM (55 9.5%)
0.5 μM (97 38.9%)
0.6 μM (90 11.4%)
2
0.4 μM (128 14.4%)
ia
6h
943 658.3
258 48.0
6
nd
nd
nd
ia
6i
5
a
nt = not tested; ia = inactive; nd = not determinable.
derivative (6g), which can be probably explained by the sterical
hindrance. A pan-agonist was obtained (6h) that activated all
classes of PPAR (58% on PPARα, 43% on PPARγ. and 25% on
PPARδ). When considering p-substituted compounds, another
dual PPARα/γ agonist was found (6i, 67% on PPARα and 84%
on PPARγ at 10 μM). Surprisingly, cyclohexyl moieties (6a−c)
did not lead to sustainable PPAR activation, indicating that this
building block is not suitable for incorporation into dual acting
sEH/PPAR modulators.
Regarding dual modulation of sEH/PPAR, we obtained two
compounds that partially activated PPARγ (6e, f) and inhibited
sEH with moderate potency (IC50 values of 304 and 73 nM,
respectively). Compound 6h inhibited sEH (IC50 = 943 nM)
and activated PPARα,γ,δ (EC50 = 6 μM on PPARα, 43% at 10
μM on PPARγ, 25% at 10 μM on PPARδ). Compound 6i
inhibited sEH (IC50 = 258 nM) and activated PPARα,γ (EC50 =
5 μM on PPARα, 84% at 10 μM on PPARγ), resulting in an
interesting compound to be evaluated in further experiments.
headgroup, known as a pharmacophore important for PPAR
dual agonistic activity, was combined with different hydro-
phobic urea derivatives in order to introduce an epoxide
mimetic.
The resulting compounds displayed high inhibition on sEH
and different patterns of PPAR agonistic activity.
This study demonstrates that the pharmacophores of PPAR
agonists and sEH inhibitors can be easily combined, resulting in
a simplified blueprint of a dual sEH/PPAR modulator. Further
in vivo pharmacological evaluation studies are needed in order
to evaluate which pattern of PPAR activation shows the most
promising profile for treatment of metabolic syndrome.
EXPERIMENTAL SECTION
■
General. All reagents and solvents were purchased from the
suppliers Alfa-Aesar GmbH & Co. KG (Karlsruhe, Germany) and
Sigma-Aldrich Chemistry GmbH (Hannover, Germany) and used
without further purification. Retention factors were determined by thin
layer chromatography with silica coated aluminum foil (particle size 60
μm) obtained from Merck KGaA (Darmstadt, Germany). Flash
chromatography was performed on packed silica columns (particle size
50 μm) from Varian Medical Systems GmbH (Darmstadt, Germany).
1H (250/400 MHz) and 13C (64 MHz) spectra were measured on AV
250 and AMX 400 nuclear magnetic resonance spectrometers from
CONCLUSION
■
This work describes the synthesis of dual sEH/PPAR
modulators as potential agents for the treatment of metabolic
syndrome. Following a combinatorial approach, an acidic
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Journal of Medicinal Chemistry
Brief Article
Bruker. Mass spectra were measured using electrospray ionization (+)
with a VG Plattform II spectrometer from Fisons. High resolution
mass spectra were measured by a MALDI LTQ Orbitrap XL
spectrometer from Thermo Scientific. All compounds were charac-
terized by NMR and MS. All final compounds had a purity of ≥95% as
determined by HPLC (LC2020, Shimadzu, Duisburg, Germany),
except for 6f (92%). Data are expressed as mean values with SE. All
IC50 and EC50 values are means with SE of the IC50 or EC50 values
obtained from measurements at five different concentrations of the
compounds in 3−5 independent experiments. IC50 and EC50 values
were determined using a sigmoidal dose response (variable slope)
equation from GraphPad Prism (GraphPad Software, LaJolla, CA)
software.
General Procedure for the Preparation of Compounds 3a−c
(o-, m-, p-). A solution of ethyl 1H-pyrrole-2-carboxylate (1) (7.2
mmol) in DCM (50 mL) was cooled to 0 °C. Tetrabutylammonium
iodide (0.22 mmol) and 50 mL of sodium hydroxide solution (50%)
were added. The mixture was stirred for 30 min at 0 °C before
(bromomethyl)benzonitrile (2) (7.9 mmol) was added. The mixture
was allowed to warm to room temperature and stirred vigorously
overnight. The reaction was quenched by adding concentrated
hydrochloric acid until a pH of 1 was reached. The aqueous layer
was extracted three times with 20 mL DCM. The collected organic
layers were washed twice with 10 mL of brine, dried over MgSO4, and
concentrated under reduced pressure. After purification by flash
chromatography (Hex:EE 0−15%), white crystals remained.
General Procedure for the Preparation of the Compounds
4a−c. A mixture of ethyl (cyanobenzyl)-1H-pyrrole-2-carboxylate
(3a−c) (5.9 mmol, 1 equiv) and Raney nickel (5.9 mmol) in 100 mL
of dry ammoniacal methanol was stirred overnight at room
temperature at H2 atmosphere under a pressure of 6 bar. The catalyst
was filtered off through Celite, and the solvent was removed under
reduced pressure to give a yellow oil.
PPAR Transactivation Assay. COS7 cells were grown in DMEM
supplemented with 10% FCS, sodium pyruvate, and penicillin/
streptomycin at 37 °C and 5% CO2. The day before transfection,
cells were seeded in 96-well plates at a density of 30000 cells per well.
Transient transfection was carried out by Lipofectamine LTX reagent
(Invitrogen) according to the manufacturer’s protocol with pFR-Luc
(Stratagene), pRL-SV40 (Promega), and the Gal4-fusion receptor
plasmids (pFA-CMV-hPPAR-LBD) of the respective subtype. Five h
after transfection, the medium was changed to DMEM without phenol
red and 10% FCS, containing 0.1% DMSO and the respective
concentrations of the test compounds.
Following overnight incubation with the test compounds, cells were
assayed for reporter gene activity using Dual-Glo luciferase assay
system (Promega, Mannheim, Germany) according to the manufac-
turer’s protocol. Luminescence was measured with a GENios Pro
luminometer (Tecan Deutschland GmbH, Crailsheim, Germany).
Each concentration of the compounds was tested in triplicate wells,
and each experiment was repeated independently at least three times.
Normalization for transfection efficacy and cell growth was done by
division of the firefly luciferase data by renilla luciferase data, resulting
in relative light units. Activation factors were obtained by dividing by
DMSO control. EC50 and standard deviation values were calculated by
mean values of at least three determinations by SigmaPlot 2001 (Systat
Software GmbH, Erkrath, Germany) using a four-parameter logistic
regression. All compounds were evaluated by comparison of the
achieved maximum effect to that of the reference compound (GW
7647 for PPARα, pioglitazone for PPARγ, and L165,041 for PPARδ
each with 1 μM).
ASSOCIATED CONTENT
■
S
* Supporting Information
Assay protocols, purity determination method, and NMR and
mass analysis data of compounds 5a−i and 6b-i. This material
is available free of charge via the Internet at http://pubs.acs.
org..
General Procedure for the Preparation of the Compounds
5a−i. A solution of ethyl 1-(aminomethyl)benzyl)-1H-pyrrole-2-
carboxylate (4a−c) (0.97 mmol) and N,N-diisopropylethylamine
(2.9 mmol) in 10 mL of dry DCM was stirred at room temperature
under argon. An isocyanate (0.97 mmol) was added, and the mixture
was stirred overnight. The solvent was evaporated under reduced
pressure. The crude product was purified by hot filtration in hexane
and recrystallized from ethanol.
AUTHOR INFORMATION
■
Corresponding Author
*E-mail: proschak@pharmchem.uni-frankfurt.de.
General Procedure to Obtain Carboxylic Acid Compounds
6a−i. The ethyl ester (5a−i) was treated with potassium hydroxide
(0.652 mmol) in a solvent mixture of THF/MeOH/H2O (1:2:2) and
accomplished under microwave irradiation at 90 °C (35 W) during 15
min. The solvent was removed under reduced pressure and the residue
solved in water. For precipitation, 1 M hydrochloric acid was added.
After filtration, the white solid was lyophilized. 1-(2-((3-
Cyclohexylureido)methyl)benzyl)-1H-pyrrole-2-carboxylic acid (6a)
was obtained from ethyl 1-(((ureido)methyl)benzyl)-1H-pyrrole-2-
carboxylate 5a (46 mg, 0.12 mmol) and yielded 25 mg (50% yield). 1H
NMR (CH3OH-d4): δ 7.19 (d, J = 7.2 Hz, 1H), 7.08−7.04 (m, 2H),
6.9 (m, 1H), 6.77 (m, 1H), 6.39 (d, J = 7.2 Hz, 1H), 6.09 (m, 1H),
5.54 (s, 2H), 4.29 (s, 2H), 3.39 (m 1H), 1.79 (m, 2H), 1.64 (m, 2H),
1.51 (m, 1H) and 1.25 (m, 5H) ppm. 13C NMR (CH3OH-d4): δ
160.0, 162.6, 137.7, 135.0, 130.1, 127.3, 126.9, 125.5, 119.9, 117.3,
110.1, 55.9, 52.6, 42.5, 34.3, 31.4, 26.2, 25.3, and 24.8 ppm. HPLC
(98% purity). HRMS: measured m/z [M + H+] 356.1967 (theoretical,
356.1969).
Author Contributions
†These authors contributed equally. The manuscript was
written through contributions of all authors. All authors have
given approval to the final version of the manuscript.
Notes
The authors declare no competing financial interest.
ACKNOWLEDGMENTS
■
This work was supported by the Deutsche Forschungsgemein-
schaft (DFG, Sachbeihilfe PR1405/2-1), LOEWE Lipid
Signaling Forschungszentrum Frankfurt (LiFF), the Oncogenic
Signaling Frankfurt (OSF), and Fonds der Chemischen
Industrie. E.B. thanks DAAD-La Caixa and R.B. the Else-
Kroner-Fresenius Foundation Graduiertenkolleg TRIP for a
̈
fellowship.
Activity Assays. sEH activity assay. For the recombinant affinity
purified sEH, we used a fluorescent-based assay19 that uses PHOME
(3-phenyl-cyano(6-methoxy-2-naphthalenyl)methyl ester-2-oxirane-
acetic acid) in a 96-well format assay to determine IC50 values.
Recombinant sEH (2 μg/well) was incubated with inhibitors for 10
min at room temperature in 25 mM Bis-Tris/HCl and 0.1 mg/mL
BSA buffer (110 μL, pH 7.0) before substrate (PHOME) was added
([S]final = 50 μM). Activity was evaluated by measuring the appearance
of the fluorescent product 6-methoxynaphthaldehyde (λem = 330 nm,
λex = 465 nm).
ABBREVIATIONS USED
■
AUDA, 12-(3-adamantan-1-yl-ureido)dodecanoic acid;
DHETs, dihydroxyeicosatrienoic acids; EETs, epoxyeicosatrie-
noic acids; FCS, fetal calf serum; LDL, low-densitiy lipoprotein;
MetS, metabolic syndrome; PPAR, peroxisome proliferator-
activated receptor; sEH, soluble epoxide hydrolase; sEHI,
soluble epoxide hydrolase inhibitor; TBAI, tetrabutyl ammo-
nium iodide; TZD, thiazolidinedione
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Journal of Medicinal Chemistry
Brief Article
(18) Pirard, B. Peroxisome Proliferator-Activated Receptors target
family landscape: a chemometrical approach to ligand selectivity based
on protein binding site analysis. J. Comput.-Aided Mol. Des. 2003, 17,
785−796.
(19) Gomez, G. A.; Morisseau, C.; Hammock, B. D.; Christianson, D.
W. Human soluble epoxide hydrolase: structural basis of inhibition by
4-(3-cyclohexylureido)-carboxylic acids. Protein Sci. 2006, 15, 58−64.
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