Synthesis and PGE2 production inhibition of s-triazine derivatives as a novel scaffold in RAW 264.7 macrophage cells

Article abstract of DOI:10.1016/j.bmcl.2014.10.031

We present the synthesis and biological evaluation of a collection of s-triazine derivatives as a novel scaffold of compounds with the capability to inhibit the PGE₂ production in LPS-induced RAW 264.7 macrophage cells. A total of 12 derivatives were synthesized and assayed for PGE₂ reduction at 10 μM concentration. Two compounds (7b and 7i) exhibiting >90% inhibition of PGE₂ production were found to have IC₅₀ values of 5.76 and 5.52 μM, respectively. They were counter screened for inhibition on COX-2 activity in a cell free assay. Specifically, compound 7i (R¹ = 4-Bn-Ph, R² = Cl, R³ = Ph, R⁵ = CO₂Me) was highly active in cells while maintaining little COX-2 inhibition (～0% at 10 μM). Molecular docking study provides the possibility that compound 7i could inhibit PGE₂ production by blocking the PGH₂ binding site of mPGES-1 instead of COX-2 enzyme. Based on this result, our synthetic efforts will focus on intensive structure-activity relationship (SAR) study of s-triazine scaffold to discovery a potential PGE₂ synthesis inhibitor.

Full text of DOI:10.1016/j.bmcl.2014.10.031

Bioorganic & Medicinal Chemistry Letters xxx (2014) xxx–xxx
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Synthesis and PGE2 production inhibition of s-triazine derivatives
as a novel scaffold in RAW 264.7 macrophage cells
Seoung Mook Kang a, , Jinsung Lee a, , Jae Ho Jin a, Minju Kim a, Sunhoe Lee a, Hwi Ho Lee b,
Ji-Sun Shin c, Kyung-Tae Lee b, , Jae Yeol Lee a,
⇑
⇑
a Research Institute for Basic Sciences and Department of Chemistry, College of Sciences, Kyung Hee University, Seoul 130-701, Republic of Korea
b Department of Life and Nanopharmaceutical Science, Kyung Hee University, 1 Hoegi-dong, Dongdaemun-gu, Seoul 130-701, Republic of Korea
c Reactive Oxygen Species Medical Research Center, School of Medicine, Kyung Hee University, Seoul 130-701, Republic of Korea
a r t i c l e i n f o
a b s t r a c t
Article history:
We present the synthesis and biological evaluation of a collection of s-triazine derivatives as a novel scaf-
Received 4 August 2014
Revised 4 October 2014
Accepted 9 October 2014
Available online xxxx
fold of compounds with the capability to inhibit the PGE2 production in LPS-induced RAW 264.7 macro-
phage cells. A total of 12 derivatives were synthesized and assayed for PGE2 reduction at 10
concentration. Two compounds (7b and 7i) exhibiting >90% inhibition of PGE2 production were found
to have IC50 values of 5.76 and 5.52 M, respectively. They were counter screened for inhibition on
COX-2 activity in a cell free assay. Specifically, compound 7i (R1 = 4-Bn-Ph, R2 = Cl, R3 = Ph, R5 = CO2Me)
M). Molecular docking
lM
l
Keywords:
was highly active in cells while maintaining little COX-2 inhibition (ꢀ0% at 10
l
Inflammation
Prostaglandin E2
Cyanuric chloride
s-Triazine
study provides the possibility that compound 7i could inhibit PGE2 production by blocking the PGH2
binding site of mPGES-1 instead of COX-2 enzyme. Based on this result, our synthetic efforts will focus
on intensive structure–activity relationship (SAR) study of s-triazine scaffold to discovery a potential
PGE2 synthesis inhibitor.
Isostere
Docking study
Ó 2014 Elsevier Ltd. All rights reserved.
PGE2 (prostaglandin E2) has long been considered the principal
prostaglandin of acute inflammation and of chronic diseases such
as rheumatoid arthritis1 and inflammatory bowel disease.2 Macro-
phages play particularly important roles in inflammation because
they produce many pro-inflammatory molecules such as PGE2.
Therefore, the pharmacological interference of PGE2 production
has been postulated as a means of alleviating a number of disease
states mediated by excessive and/or protracted macrophage acti-
vation. As PGE2 is produced in relative abundance by COX-2 (cyclo-
oxygenase-2) action, selective COX-2 inhibitors such as celecoxib
and rofecoxib cause PGE2 levels to drop both absolutely and rela-
tive to the other prostaglandins. However, coxib drugs such as Bex-
traÒ and VioxxÒ were withdrawn from the market in 2004 and
2005, respectively, because they excessively increased the risk of
heart attacks and strokes with high doses and chronic usage.3
Celecoxib (Celebrex)Ò is the only COX-2 inhibitor available in the
United States. Therefore, the exploration of alternative pharmaco-
logical approaches leading to safer anti-inflammatory drugs
that do not alter COX-2 activity is of urgent need. One promis-
ing approach to circumvent COX-related side effects while
maintaining anti-inflammatory efficacy is the interference
with the microsomal prostaglandin E2 synthase (mPGES)-1. This
mPGES-1 enzyme, a downstream PG synthase, specifically cata-
lyzes the biosynthesis of COX-2-derived PGE2 from PGH2 (prosta-
glandin H2)4 and thus, its inhibition would ideally not affect the
formation of house-keeping PGs. Therefore, the small molecule
inhibitors of mPGES-1 would serve as a beneficial anti-inflamma-
tory therapy with reduced side effects.5–11
Cyanuric chloride, the alternate name 2,4,6-trichloro-s-triazine,
is the trimer of cyanogen chloride (ClCN). It is the main precursor
to the popular triazine-class herbicides and polymer photostabilis-
ers12 and has been widely used in organic reactions.13–15 Such
diverse organic reactions simply rely on the easy displacement of
the chloride with diverse nucleophiles. In addition, the substituted
s-triazine class demonstrates wide biological applications—includ-
ing antimicrobial, antituberculosis, anticancer, antiviral and
antimalarial.16,17
Based on this privileged s-triazine scaffold in the field of medic-
inal chemistry, we decided to embark on the synthesis of novel s-
triazine derivatives as isosteric analogs of pirinixic acids, which
are recently reported as dual mPGES-1/5-LO (5-lipoxygenase)
inhibitors,10,18–20 for the discovery of novel anti-inflammation
agents as shown in Figure 1. In this letter, we report the synthesis
of a series of s-triazine derivatives and the inhibition of both the
⇑
Corresponding authors. Tel.: +82 2 961 0726; fax: +82 2 966 3701.
E-mail addresses: ktlee@khu.ac.kr (K.-T. Lee), ljy@khu.ac.kr (J.Y. Lee).
The contributions of the authors are equal.
http://dx.doi.org/10.1016/j.bmcl.2014.10.031
0960-894X/Ó 2014 Elsevier Ltd. All rights reserved.
Please cite this article in press as: Kang, S. M.; et al. Bioorg. Med. Chem. Lett. (2014), http://dx.doi.org/10.1016/j.bmcl.2014.10.031

2
S.M. Kang et al. / Bioorg. Med. Chem. Lett. xxx (2014) xxx–xxx
Figure 1. The replacement of pyrimidine ring with s-triazine ring as a privileged scaffold.
PGE2 production in LPS (lipopolysaccharides)-induced RAW 264.7
macrophage cells and the COX-2 enzyme in cell-free assay. Addi-
tionally, molecular docking simulation was performed to assess vir-
tually whether the activity of potent compound would be related
with the binding interaction with mPGES-1 enzyme or not.
The presented compounds (7a–j, 8 and 9) were generally syn-
thesized based on the modified method described by Werz’s group
as shown in Scheme 1.18–20 First, cyanuric chloride (1) was treated
with the respective primary amine building block (R1ANH2,
1.2 equiv) in acetone to provide the corresponding 2-amino-4,6-
dichloro-s-triazine (3b–f) via a nucleophilic substitution of one
chlorine. In the case of 2-amino-4-chloro-6-methyl-s-triazine
(3a), cyanuric chloride (1) was treated with CH3MgBr (1.2 equiv)
in Et2O–CH2Cl2 solution to provide 2,4-dichloro-6-methyl-s-tri-
azine (2),21 which was reacted with 4-isopropylphenylamine
(1.2 equiv) to afford 2-amino-4-chloro-6-methyl-s-triazine 3a. Sec-
ondly, commercially available 2-phenylacetate 4a (or 2-naphthyl
Initially, cell viability was assessed with MTT assay based on the
experimental procedures described previously.24 Most synthetic
compounds except for compound 8 did not exhibit any cytotoxic
effects against RAW 264.7 cells at concentrations above 10 lM in
the presence or absence of LPS, indicating that their suppressive
effects on PGE2 production could not be attributable to non-spe-
cific cytotoxicity. Therefore, most compounds were screened for
their ability to inhibit PGE2 production in LPS-induced RAW
264.7 cells at 10
lM concentrations (@ 1
lM for compound 8
because of cytotoxicity) using NS398 (@ 3
lM) as a positive con-
trol. PGE2 concentration in the medium was measured using EIA
kit for PGE2 according to the manufacturer’s recommendation.25
Active compounds exhibiting >90% inhibition of PGE2 production
were pushed forward for IC50 determinations. All experiments
were carried out at least twice and they gave similar results. Their
activities are summarized as both percentage inhibitions and IC50
values of PGE2 production in Table 1 with NS398 incorporated as
a positive control in both PGE2 and COX-2 assays.
Among the synthetic s-triazine derivatives, at a glance, most
compounds (7a–j except 7d) which have ester groups as the R5
substituent (i.e., R4AC@O in Scheme 1) generally showed strong
inhibitory activities (72–91% inhibition) of PGE2 production as
shown in Table 1. The compound 7d showed moderate activity
(43% inhibition) despite of having the same ester group. This result
may imply that the character of R1 substituent is related with the
bioactivity of s-triazine derivative, which effect is discussed below.
While s-triazine derivatives (8 and 9) with amide or free carboxylic
for 4b–c) was
dition.22 The resulting
with KSAc as a nucleophile in acetone, and then followed by hydro-
lysis with NaOEt/MeOH to yield
-mercaptoacetate 6a (or 6b–c).23
Finally, the nucleophilic substitution of 6-chloro-s-triazine 3a–f
with the corresponding -mercaptoacetate 6a–c in K2CO3 in DMF
a-brominated with AIBN and NBS under reflux con-
a
-bromoacetate 5a (or 5b–c) was treated
a
a
resulted in the formation of target compounds 7a–j. The treatment
of ester 7a with NH4OH in MeOH was converted into the corre-
sponding primary amide 8. The ester 7f was hydrolyzed with
KOH in THF–H2O to give the desired carboxylic acid 9.
O
H
N
N
S
NH2
R1-NH2 (1.2 equiv), acetone, rt, 1-2 h
N
N
N
N
CH3
R1-NH2
(1.2 equiv)
H
N
Cl
N
Cl
Cl
N
Cl
N
Cl
8
(51%)
R1
MeMgBr (1.2 equiv)
N
N
N
N
N
N
Et2O-CH2Cl2,
0 oC to rt, 3 h
acetone, rt, 1 h
NH4OH
MeOH, rt, 48 h
7a
Cl
CH3
R2
for
: R1 = 4-isopropylphenyl, R2 = Me (85%)
1
2
3a
(51%)
: R1 = 4-isopropylphenyl, R2 = Cl (87%)
: R1 = 4-cyclohexylphenyl, R2 = Cl (92%)
3b
3c
3d
O
H
N
K2CO3
: R1 = 4-biphenylyl,
R2 = Cl (87%)
N
S
R1
OR4
: R1 = 4-phenoxyphenyl, R2 = Cl (80%)
3e
DMF, rt,
1-4 h
R3
N
: R1 = 4-benzylphenyl,
R2 = Cl (80%)
3f
R2
7a-j (30-62%)
i) KSAc (1.1 equiv)
acetone, rt,
20-30 min
O
O
O
Br
HS
OR4
OR4
OR4
NBS (1.2 equiv), AIBN
CHCl3, reflux, 3-5 h
R3
R3
R3
KOH
THF-H2O, rt, 24 h
for 7f
ii) NaOR4, R4OH, rt,
20-30 min
4a: R3 = 1-phenyl, R4 = Me
4b: R3 = 1-naphthyl, R4 = Me
4c: R3 = 1-naphthyl, R4 = Et
5a (90%)
5b (81%)
5c (75%)
6a (83%)
6b (80%)
6c (86%)
O
H
N
N
S
OH
N
Cl
9
(35%)
Scheme 1. Synthesis of s-triazine derivatives.
Please cite this article in press as: Kang, S. M.; et al. Bioorg. Med. Chem. Lett. (2014), http://dx.doi.org/10.1016/j.bmcl.2014.10.031

S.M. Kang et al. / Bioorg. Med. Chem. Lett. xxx (2014) xxx–xxx
3
Table 1
Inhibition of PGE2 production and COX-2 inhibition of s-triazine derivatives
H
N
N
S
R5
R1
N
N
R3
R2
Entry
R1
R2
R3
R5
Yielda
(%)
Viability
M)
PGE2
%inhibition
@10
COX-2
(
l
b
IC50
M)
%inhibition
l
M
(
l
@ 10
lM
7a
7b
7c
7d
7e
7f
7g
7h
7i
4-Isopropylphenyl
4-Isopropylphenyl
4-Isopropylphenyl
4-Cyclohexylphenyl
4-Biphenylyl
Me
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Cl
Ph
Ph
CO2Me
CO2Me
CO2Me
CO2Et
CO2Me
CO2Et
CO2Me
CO2Me
CO2Me
CO2Et
CONH2
CO2H
51
51
42
62
55
33
30
53
37
55
51
35
>10
>10
>10
>10
>10
>10
>10
>10
>10
>10
<10
>10
85.65
91.81
88.69
43.68
82.67
78.56
85.77
72.46
90.81
72.60
5.76
31.8
1-Naphthyl
1-Naphthyl
Ph
1-Naphthyl
Ph
1-Naphthyl
Ph
1-Naphthyl
Ph
4-Biphenylyl
4-Phenoxyphenyl
4-Phenoxyphenyl
4-Benzylphenyl
4-Benzylphenyl
4-Isopropylphenyl
4-Biphenylyl
5.52
n.i.c
7j
8
Cl
Me
Cl
26.50 (1)d
7.77
n.d.
n.d.
7.0 nM
9
1-Naphthyl
NS398e
99.42 (3)f
1.8 lM (IC50)
a
b
c
d
e
f
Isolated yield of final step for each compound.
IC50 value of PGE2 production inhibition for active compounds and NS398 as a positive control in LPS-induced RAW 264.7 macrophage cells.
n.i. = no inhibition (ꢀ0%).
% inhibition at 1 lM concentration due to its cytotoxicity at 10 lM concentration.
Positive control compound for assay of PGE2 production inhibition.
% inhibition at 3 M concentration.
l
acid group as R5 substituent showed the poor reduction of cellular
PGE2 levels when compared to the activities of the parent com-
pounds (7a and 7f). In particular, s-triazine derivative 9 exhibited
much reduced bioactivity (7%), possibly due to its reduced ability
to permeate the cellular membrane. Generally, the compounds
(7b, 7e, 7g and 7i) with phenyl ring as R3 substituent were slightly
more active than the corresponding compounds with 1-naphthyl
ring (7c, 7f, 7h and 7j). The introduction of methyl group as the
R2 substituent of s-triazine ring was almost without effect (7a vs
7b). Finally, the relative profile with respect to R1 substituent
was found to be 4-isopropylphenyl (7b) P 4-benzylphenyl
(7i) > 4-phenoxyphenyl (7g) P 4-biphenylyl (7e) >> 4-cyclohexyl-
phenyl (7d). Overall, the optimal substituents in s-triazine deriva-
tive for inhibitory activity of PGE2 production in LPS-induced RAW
264.7 cells are found to be 4-isopropylphenyl (or 4-benzylphenyl)
ring at R1, chloro atom at R2, phenyl ring at R3 and ester group at R5
substituent in this series. Consequentially, compounds 7b and 7i
with these optimal substituent profiles exhibited >90% inhibition
of PGE2 production, which is comparable to the result of the posi-
tive inhibitor, NS398. Further test showed that 7b and 7i sup-
pressed the production of PGE2 with IC50 values of 5.76 and
Figure 2. Inhibitory effects of compounds 7b and 7i on LPS-induced PGE2
production in RAW 264.7 macrophages. NS398 as a positive control was tested at
3 lM concentration. Data are presented as the means SD of three independent
experiments.
5.52 lM, respectively, as shown in Figure 2.
In addition, we further investigated whether the inhibitory
activity of active compounds 7b and 7i on LPS-induced PGE2 pro-
duction is related with COX-2 inhibitory activity or not. Two com-
pounds were tested for the inhibitory activities against human
5-lipoxygenase, and PGE2 synthases (PGES),27 we decided to virtu-
ally investigate the possibility of compounds 7b and 7i to inhibit
microsomal prostaglandin
molecular docking study because it could be regarded as an bioiso-
stere of pirinixic acid derivatives, as dual PPAR activators and
E synthase-1 (mPGES-1) by using
recombinant COX-2 inhibition at each 10
lM concentration using
a/c
enzyme immunoassay (EIA) kit (kit catalog number 560101, Cay-
man Chemical, Ann Arbor, MI, USA) in a cell free assay. Interest-
ingly, 7b exhibited 31% inhibition on COX-2 activity, whereas 7i
dual 5-LO/mPGES-1 inhibitors as shown in Figure 1.20 The docking
studies were performed using Molegro Virtual Docker (MVD)
2013.6.0.1 for Windows28 and the recent human mPGES-1 X-ray
structure (PDB code: 4AL0).29 First, we identified one cavity with
a volume 59 Å located in close proximity to GSH (glutathione) as
a cofactor by the same software, which can automatically detect
cavities from protein surfaces. The putative active site of the
enzyme was defined to include residues within a 15 Å radius to
this cavity. The docking wizard of 2013.6.0.1 was used to dock all
compounds as shown in Figure 3 for both docking validation and
data comparison. The most stable docking pose was selected
did not show any inhibitory activity at 10 lM concentration. These
overall results implicate that compound 7i may have particularly
the capacity to reduce cellular PGE2 levels by inhibiting other
PGE2 biosynthesis-related enzymes instead of COX-2 enzyme
when compared to our assay data of NS398 (selective COX-2 inhib-
itor; IC50 = 1.8/220 lM against COX-2/1 enzymes; IC50 = 7.0 nM
against PGE2 production) as a positive control.26 Among other
enzymes involved in PGE2 biosynthesis such as phospholipases,
Please cite this article in press as: Kang, S. M.; et al. Bioorg. Med. Chem. Lett. (2014), http://dx.doi.org/10.1016/j.bmcl.2014.10.031

4
S.M. Kang et al. / Bioorg. Med. Chem. Lett. xxx (2014) xxx–xxx
O
O
Before starting with our active compounds 7b and 7i, molecular
docking study with three reported mPGES-1 inhibitors (MK886,
and two compounds arbitrarily named JMC-1 and JMC-2 for our
convenience)20 was performed in the presence or absence of gluta-
thione (GSH) as a cofactor in order to investigate the effect of GSH
on this docking study. As shown in Table 2, the appropriate corre-
lation between real IC50 values and MolDock Scores was found only
in the presence of GSH when considered for the data of JMC-1, 2
H
N
H
N
N
S
N
S
OCH3
OCH3
N
N
N
N
Cl
Cl
7b
7i
O
O
H
N
H
N
N
S
N
S
OH
OC2H5
Hexyl
and MK886 (IC50 = 5.1, 5.6 and 2.1
l
M vs À140, À131 and À166,
N
N
N
respectively). In addition, NS398 as a selective COX-2 inhibitor
showed the least MolDock Score of À95 as predicted. Under the
same condition, our active compound 7i was found to dock into
the active site of mPGES-1 with high MolDock Score of À145 com-
parable to that of JMC-1 and also be slightly higher than 7b. Based
Cl
Cl
JMC-1
JMC-2
O
O
S
S
N
on little effect of 7i on COX-2 activity at 10 lM concentration, this
HN
docking result provides the possibility that our active compound 7i
could inhibit the PGE2 production by blocking the active site (i.e.,
the binding site for PGH2) of mPGES-1 enzyme instead of COX-2
enzyme. Figure 4 shows the docked structure of active compound
7i binding with mPGES-1. Two oxygen atoms of ester group in
compound 7i form two hydrogen bonds with Gly35 of chain A
and Tyr130 of chain B, and three nitrogen atoms of s-triazine ring
form strongly four hydrogen bonds with the cofactor (GSH). This
strong interaction of s-triazine ring of 7i with GSH mainly contrib-
utes to its high MolDock Score in this docking study. Due to its high
O
CO2H
Cl
NO2
MK886
NS398
Figure 3. Chemical structures of selected compounds for molecular docking study.
according to the best MolDock Score conformation predicted by
the MVD scoring function in Table 2.
Table 2
Docking results of compounds 7b, 7i, reported mPGES-1 inhibitors (JMC-1 and 2) and NS398 into the active sites of mPGES-1
Compound
Human COX-2
(% inhibition @ 10 lM, or IC50)
Human mPGES-1
MolDock Scorea (PDB code: 4AL0)
(IC50: l
M)b
In the presence GSH
In the absence of GSH
7b
7i
31.8
ꢀ0
n.d.c
n.d.
5.1
5.6
2.1
À132
À145
À140
À131
À166
À95
À117
À138
À156
À122
À153
À83
JMC-1b
JMC-2b
MK886b,d
NS398f
67.5
38.3
58
l
Me (IC50
M (IC50
)
)
1.8
l
n.d.
a
b
c
d
e
f
MolDock Score in 2013.6.0.1 for Windows.
Selected compounds and their data from the reported literature: Koeberle, A., et al. J. Med. Chem. 2008, 51, 8068.
n.d. = not determined.
mPGES-1 inhibitor was docked for the relative comparison.
Reported data: Koeberle, A., et al. Eur. J. Pharmacol. 2009, 608, 84.
Selective COX-2 inhibitor was docked for the relative comparison.
Figure 4. Three dimensional mode of compound 7i in the active site of mPGES-1 (PDB code: 4ALO). 3D docking mode of compound 7i into the active site of mPGES-1. GSH
(glutathione) as a cofactor is depicted as ball and stick model. The key amino acids of chain A and B participating in hydrogen bonds are depicted as yellow stick model.
Hydrogen bonds are shown in blue dotted line. The only polar hydrogens are shown for the simplicity.
Please cite this article in press as: Kang, S. M.; et al. Bioorg. Med. Chem. Lett. (2014), http://dx.doi.org/10.1016/j.bmcl.2014.10.031

S.M. Kang et al. / Bioorg. Med. Chem. Lett. xxx (2014) xxx–xxx
5
10. Hanke, T.; Lamers, C.; Gomez, R. C.; Schneider, G.; Werz, O.; Schubert-Zsilavecz,
M. Bioorg. Med. Chem. Lett. 2014, 24, 3757.
11. Bahia, M. S.; Katare, Y. K.; Silakari, O.; Vyas, B.; Silakari, P. Med. Res. Rev. 2014,
34, 825.
12. Hollink, E.; Simanek, E. E.; Bergbreiter, D. E. Tetrahedron Lett. 2005, 46, 2005.
13. Li, W. L.; Luo, Q. Y.; Yan, F. L. Chin. Chem. Lett. 2011, 22, 811.
14. Venkataraman, K.; Wagle, D. R. Tetrahedron Lett. 1979, 20, 3037.
15. Encyclopedia of Reagents for Organic Synthesis; Paquette, L. A., Crich, D., Fuchs, P.
L., Molander, G. A., Eds.; John Wiley & Sons, 2004; Vol. 4, pp 2874–2878.
16. Patel, R. V.; Kumari, P.; Rajani, D. P.; Pannecouque, C.; De Clercq, E.; Chikhalia,
K. H. Future Med. Chem. 2012, 4, 1053.
reactivity and binding characteristic towards various enzymes, the
s-triazine unit has attracted attention for the construction of chem-
ical libraries by combinatorial approaches.30 In addition, this bind-
ing mode of 7i is also in agreement with some of the key
interactions reported for the putative mPGES-1 inhibitor.29
In conclusion, a series of novel s-triazine compounds as isoster-
es of pirinixic acid derivatives have been synthesized via the
replacement strategy of pyrimidine ring of with s-triazine ring
and their effects on PGE2 production in LPS-induced RAW 264.7
cells were evaluated. Among them, compound 7i exhibited the
17. Shah, D. R.; Modh, R. P.; Chikhalia, K. H. Future Med. Chem. 2014, 6, 463. and
references cited therein.
18. Hanke, T.; Dehm, F.; Liening, S.; Popella, S. D.; Maczewsky, J.; Pillong, M.;
Kunze, J.; Weinigel, C.; Barz, D.; Kaiser, A.; Wurglics, M.; Lämmerhofer, M.;
Schneider, G.; Sautebin, L.; Schubert-Zsilavecz, M.; Werz, O. J. Med. Chem. 2013,
56, 9031.
19. Hieke, M.; Greiner, C.; Thieme, T. M.; Schubert-Zsilavecz, M.; Werz, O.; Zettl, H.
Bioorg. Med. Chem. Lett. 2011, 21, 1329.
high inhibition on PGE2 production with an IC50 value of 5.52 lM
and was non-cytotoxic at the tested concentration. Docking study
revealed that 7i could bind favorably to mPGES-1 enzyme and will
serve as an important template for future chemical modification.
Future studies will address on the intensive structure–activity rela-
tionship (SAR) study of 7i for the discovery of a potent PGE2 syn-
thesis inhibitor with no effect on COX-2 enzyme.
20. Koeberle, A.; Zettl, H.; Greiner, C.; Wurglics, M.; Schubert-Zsilavecz, M.; Werz,
O. J. Med. Chem. 2008, 51, 8068.
21. Smith, A. L.; D’Angelo, N. D.; Bo, Y. Y.; Booker, S. K.; Cee, V. J.; Herberich, B.;
Hong, F. T.; Jackson, C. L.; Lanman, B. A.; Liu, L.; Nishimura, N.; Pettus, L. H.;
Reed, A. B.; Tadesse, S.; Tamayo, N. A.; Wurz, R. P.; Yang, K.; Andrews, K. L.;
Whittington, D. A.; McCarter, J. D.; Miguel, T. S.; Zalameda, L.; Jiang, J.;
Subramanian, R.; Mullady, E. L.; Caenepeel, S.; Freeman, D. J.; Wang, L.; Zhang,
N.; Wu, T.; Hughes, P. E.; Norman, M. H. J. Med. Chem. 2012, 55, 5188.
22. Neto, B. A. D.; Lapis, A. A. M.; Bernd, A. B.; Russowsky, D. Tetrahedron 2009, 65,
2484.
Acknowledgments
This research was supported by Basic Science Research Program
through the National Research Foundation of Korea (NRF) funded
by the Ministry of Education (NRF-2013R1A1A2058915).
23. Ayscough, A.; Owen, D. R.; Meo, P.; Pearson, D. J.; Walker, Y.; Boyce, R. J.;
Zuccotto, F. WO 2007036730 A1, 2007.
24. Won, J. H.; Im, H. T.; Kim, Y. H.; Yun, K. J.; Park, H. J.; Choi, J. W.; Lee, K. T. Br. J.
Pharmacol. 2006, 148, 216.
References and notes
25. Mossman, T. J. Immunol. Methods 1983, 65, 55. The RAW 264.7 macrophage cell
line was obtained from the Korea Cell Line Bank (Seoul, Korea). Cells were
grown at 37 °C in DMEM supplemented with 10% FBS, penicillin (100 units/ml),
1. Akaogi, J.; Nozaki, T.; Satoh, M.; Yamada, H. Endocr. Metab. Immun. Disord. Drug
Targets 2006, 6, 383.
2. Blázovics, A.; Hagymási, K.; Prónai, L. Orv. Hetil. 2004, 145, 2523.
3. McGettigan, P.; Henry, D. JAMA 2006, 296, 1633.
4. Koeberle, A.; Rossi, A.; Zettl, H.; Pergola, C.; Dehm, F.; Bauer, J.; Greiner, C.;
Reckel, S.; Hoernig, C.; Northoff, H.; Bernhard, F.; Dötsch, V.; Sautebin, L.;
Schubert-Zsilavecz, M.; Werz, O. J. Pharmacol. Exp. Ther. 2010, 332, 840.
5. Riendeau, D.; Aspiotis, R.; Ethier, D.; Gareau, Y.; Grimm, E. L.; Guay, J.; Guiral, S.;
Juteau, H.; Mancini, J. A.; Methot, N.; Rubin, J.; Friesen, W. Bioorg. Med. Chem.
Lett. 2005, 15, 3352.
and streptomycin sulfate (100 lg/ml) in a humidified 5% CO2 atmosphere. Cells
were incubated with the tested samples at increasing concentrations or
positive chemical (NS398) and then stimulated with LPS 10 ng/ml for the
indicated time. PGE2 concentration in the medium was quantified using EIA
kits (R&D Systems, Minneapolis, MN, USA).
26. Kim, K. J.; Choi, M. J.; Shin, J. S.; Kim, M.; Choi, H. E.; Kang, S. M.; Jin, J. H.; Lee,
K.-T.; Lee, J. Y. Bioorg. Med. Chem. Lett. 2014, 24, 1958.
27. Smith, W. L. Biochem. J. 1989, 259, 315.
6. Cote, B.; Boulet, L.; Brideau, C.; Claveau, D.; Ethier, D.; Frenette, R.; Gagnon, M.;
Giroux, A.; Guay, J.; Guiral, S.; Mancini, J.; Martins, E.; Masse, F.; Methot, N.;
Riendeau, D.; Rubin, J.; Xu, D.; Yu, H.; Ducharme, Y.; Friesen, R. W. Bioorg. Med.
Chem. Lett. 2007, 17, 6816.
7. Giroux, A.; Boulet, L.; Brideau, C.; Chau, A.; Claveau, D.; Cote, B.; Ethier, D.;
Frenette, R.; Gagnon, M.; Guay, J.; Gurial, S.; Mancini, J. A.; Martins, E.; Masse,
F.; Methot, N.; Riendeau, D.; Rubin, J.; Xu, D.; Yu, H.; Ducharme, Y.; Friesen, R.
W. Bioorg. Med. Chem. Lett. 2009, 19, 5837.
28. MVD 2013.6.0.1 for Windows in Molegro ApS: With this purpose, crystal
structures of human mPGES-1 (PDB code: 4AL0) was obtained from the Protein
Data Bank in order to prepare the protein for docking studies. Docking
procedure was followed using the standard protocol implemented in MVD
2013.6.0.1 and the geometry of resulting complexes was studied using the
MVD’s Pose Viewer utility.
29. Sjögren, T.; Nord, J.; Ek, M.; Johansson, P.; Liu, G.; Geschwindner, S. Proc. Natl.
Acad. Sci. U.S.A. 2013, 110, 3806.
8. Hamza, A.; Zhao, X.; Tong, M.; Tai, H. H.; Zhan, C. G. Bioorg. Med. Chem. 2011, 19,
6077.
9. Lee, K.; Pham, V. C.; Choi, M. J.; Kim, K. J.; Lee, K.-T.; Han, S. G.; Yu, Y. G.; Lee, J. Y.
Bioorg. Med. Chem. Lett. 2013, 23, 75.
30. Khersonsky, S. M.; Jung, D.-W.; Kang, T.-W.; Walsh, D. P.; Moon, H.-S.; Jo, H.;
Jacobsen, E. M.; Shetty, V.; Neubert, T. A.; Chang, Y.-T. J. Am. Chem. Soc. 2003,
125, 11804.
Please cite this article in press as: Kang, S. M.; et al. Bioorg. Med. Chem. Lett. (2014), http://dx.doi.org/10.1016/j.bmcl.2014.10.031