Thiazolidine-2,4-diones derivatives as PPAR-γ agonists: Synthesis, molecular docking, in vitro and in vivo antidiabetic activity with hepatotoxicity risk evaluation and effect on PPAR-γ gene expression

Article abstract of DOI:10.1016/j.bmcl.2014.05.034

A library of conjugates of chromones and 2,4-thiazolidinedione has been synthesized by Knoevenagel condensation followed by reduction using hydrogen gas and Pd/C as a catalyst. Compounds 5c and 5e were most effective in lowering the blood glucose level comparable to standard drug pioglitazone. Compound 5e exhibited potent PPAR-γ transactivation of 48.72% in comparison to pioglitazone (62.48%). All the molecules showed good glide score against the PPAR-γ target in molecular docking study. PPAR-γ gene expression was significantly increased by compound 5e (2.56-fold) in comparison to standard drug pioglitazone. Compounds 5e and 5c did not cause any damage to the liver and may be considered as promising candidates for the development of new antidiabetic agents.

Full text of DOI:10.1016/j.bmcl.2014.05.034

Bioorganic & Medicinal Chemistry Letters 24 (2014) 3034–3042
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Thiazolidine-2,4-diones derivatives as PPAR-c agonists: Synthesis,
molecular docking, in vitro and in vivo antidiabetic activity with
hepatotoxicity risk evaluation and effect on PPAR-
c
gene expression
a
b
c
Syed Nazreen a, Mohammad Sarwar Alam a, , Hinna Hamid , Mohammad Shahar Yar , Abhijeet Dhulap ,
Perwez Alam d, M. A. Q. Pasha d, Sameena Bano a, Mohammad Mahboob Alam a, Saqlain Haider a,
Chetna Kharbanda a, Yakub Ali a, K. K. Pillai e
⇑
a Department of Chemistry, Faculty of Science, Jamia Hamdard (Hamdard University), New Delhi 110 062, India
b Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Jamia Hamdard (Hamdard University), New Delhi 110 062, India
c CSIR Unit for Research and Development of Information Products, Pune 411 038, India
d Functional Genomics Unit, CSIR-Institute of Genomics & Integrative Biology, New Delhi, India
e Department of Pharmacology, Faculty of Pharmacy, Jamia Hamdard (Hamdard University), New Delhi 110 062, India
a r t i c l e i n f o
a b s t r a c t
Article history:
A library of conjugates of chromones and 2,4-thiazolidinedione has been synthesized by Knoevenagel
condensation followed by reduction using hydrogen gas and Pd/C as a catalyst. Compounds 5c and 5e
were most effective in lowering the blood glucose level comparable to standard drug pioglitazone.
Received 2 December 2013
Revised 21 April 2014
Accepted 12 May 2014
Available online 22 May 2014
Compound 5e exhibited potent PPAR-
(62.48%). All the molecules showed good glide score against the PPAR-
study. PPAR- gene expression was significantly increased by compound 5e (2.56-fold) in comparison
c
transactivation of 48.72% in comparison to pioglitazone
c
target in molecular docking
c
Keywords:
2,4-Thiazolidinedione
Chromone
to standard drug pioglitazone. Compounds 5e and 5c did not cause any damage to the liver and may
be considered as promising candidates for the development of new antidiabetic agents.
Ó 2014 Elsevier Ltd. All rights reserved.
Antidiabetic activity
PPAR-
c
Hepatotoxicity
Type 2 diabetes is a complex metabolic syndrome resulting in
adipose tissues and intestine. These are encoded by PPAR-
c gene
high blood glucose due to impaired insulin action, which in turn
stimulates glucose uptake in peripheral tissues such as muscle
and fat. In normal humans, up to 80% of insulin-stimulated glucose
disposal occurs in skeletal muscle, a major site of insulin resistance
in type 2 diabetes.1,2
in humans. Although TZDs are strong and specific activators of
PPAR-c capable of ameliorating diabetes mellitus by improving
insulin resistance without inducing hypoglycaemia,18 they are
associated with side effects viz. weight gain, hepatotoxicity and
fluid retention.19
Recently, chemistry of 2,4-thiazolidinediones (TZDs) has
attracted attention as they have been found to exhibit several bio-
logical activities,3 such as antihyperglycemic,4 anti-inflammatory,5
antimalarial,6 antioxidant,7 antitumor,8 cytotoxic,9 antimicrobial,10
and antiproliferative.11 Thiazolidinediones are high-affinity ligands
Chromones are a group of naturally occurring compounds found
in fruits and vegetables.20,21 They are safe and are associated with
low mammalian toxicity, making them excellent chemopreventive
agents.22 They are reported to exert an antihyperglycemic effect by
promoting peripheral utilization of glucose or enhancing insulin
sensitivity in diabetic animals.23 Considering the biological impor-
tance of thiazolidinediones and chromones as antidiabetic agents,
we have conjugated these two important ligands under one con-
struct through a methylene linkage. We herein report the synthesis
of thiazolidinedione and chromone based conjugates and their
molecular docking studies and evaluation of their in vivo antidia-
of peroxisome proliferator activated receptor-c 12
PPAR-c, a
.
member of a large family of ligand-activated nuclear hormone
receptors is an important drug target for regulating glucose
metabolism. PPAR-
muscle and hepatic tissues13,14 thereby improving plasma glucose
levels effectively.15–17 PPAR-
exists in two forms mainly PPAR- 1,
2, found in
c increases insulin sensitivity at adipose,
c
c
found in all the tissues except muscles and PPAR-
c
betic activity with hepatotoxicity risk evaluation, in vitro PPAR-
activity and also the effect on the PPAR- gene expression.
c
c
Villsmeyer Hack reaction of substituted o-hydroxy acetoph-
enones (1a–k) in presence of DMF and POCl3 yielded 3-formyl
⇑
Corresponding author. Tel.: +91-9717927759.
E-mail address: msalam5555@gmail.com (M.S. Alam).
http://dx.doi.org/10.1016/j.bmcl.2014.05.034
0960-894X/Ó 2014 Elsevier Ltd. All rights reserved.

S. Nazreen et al. / Bioorg. Med. Chem. Lett. 24 (2014) 3034–3042
3035
OH
O
S
a
O
H
N
H
O
R
R
O
(2a-k)
O
O
(3)
(1a-k)
b
O
O
O
S
NH
R
O
(4a-k)
c
O
O
O
O
O
O
S
S
NH
NH
R
R
O
O
By product (6a-j)
Desired product (5a-k)
Scheme 1. Reagents and conditions:(a) DMF, POCl3 , 0-(À5)°C;(b) sodium acetate, acetic acid, 120–140 °C; (c) Pd/C, H2 gas, 30 psi, rt, 10–12 h.
Table 1
Physical data of the synthesized compounds
S.No.
Reactant (4)
Desired product (5)
yield (%)
50
By product (6)
Yield (%)
O
O
O
O
O
O
S
S
S
NH
NH
NH
30
20
a
O
O
O
O
O
O
O
O
O
O
O
O
O
S
S
S
NH
NH
b
c
65
70
NH
O
O
O
O
O
O
O
O
O
O
O
S
S
S
NH
NH
NH
20
O
O
O
O
O
S
O
O
O
O
O
O
O
O
O
O
S
S
NH
NH
NH
d
e
f
65
60
65
25
20
21
Br
Cl
F
Br
Br
Cl
F
O
O
O
O
O
O
O
O
O
O
O
O
S
S
S
NH
NH
NH
Cl
O
O
O
O
O
O
O
O
O
O
O
O
S
S
S
NH
NH
NH
F
O
O
O
O
O
O
(continued on next page)

3036
S. Nazreen et al. / Bioorg. Med. Chem. Lett. 24 (2014) 3034–3042
Table 1 (continued)
S.No.
g
Reactant (4)
Desired product (5)
yield (%)
70
By product (6)
Yield (%)
O
O
F
O
O
F
O
S
S
NH
NH
—
O
O
O
S
O
O
O
O
O
O
O
O
S
NH
NH
h
75
64
—
O
O
O
Cl
Cl
Cl
O
O
O
O
O
O
S
S
S
i
21
25
NH
NH
NH
Cl
Cl
Cl
O
O
O
O
O
O
O
O
O
O
O
S
S
S
NH
NH
NH
j
55
75
O
O
O
O
O
O
O
O
O
O
NH
O
NH
O
O
O
S
S
k
—
Cl
Cl
O
O
O
1H NMR, 13C NMR and mass spectral data were found to be
Table 2
Docking score and ADME prediction of the compounds (5a–k) and (6a–j)
in agreement with the proposed structures of all the newly
synthesized compounds (Supplementary data). Formation of
chromonyl-2,4-thiazolidinediones (4a–k) by the Knoevenagel
condensation was confirmed by the appearance of exocyclic ole-
finic proton as a singlet in the range of d 7.57–7.79 in the 1H
NMR spectra of these compounds. Reduction of compounds
(4a–k) with Pd/C resulted in the hydrogenation of the double bond
between TZD and chromone ring leading to the formation of
compounds (5a–k) as well as hydrogenation of both the double
bonds, that is double bond between TZD and chromone ring and
olefinic bond of chromone ring gave compounds (6a–j). Formation
of compounds (5a–k) was confirmed by the disappearance of the
singlet of exocyclic olefinic proton as seen in the 1H NMR spectra
of (4a–k) and the appearance of a characteristic pattern of three
double doublets at d 4.81–4.86, 3.28–3.32 and 2.70–2.85 in the
1H NMR spectra of (5a–k) due to the newly generated methine
and methylene protons, respectively. The formation of compounds
(6a–j) was confirmed by the absence of singlet of the olefinic
proton of chromone ring at d 8.31–8.48 and appearance of double
doublet of methylene protons of chromone ring at d 4.30–4.66,
double doublet of methine protons of TZD ring at d 4.16–4.37
and a multiplet in the region d 2.50–2.62 in the 1H NMR spectra.
The double doublets of methylene protons that appeared in the
1H NMR spectra of compounds (5a–k) were not seen in the
1H NMR spectra of (6a–j). Instead, a multiplet in the region d
1.34–2.52 was observed for these protons confirming reduction
of both the double bonds in these compounds (Table 1).
S.No.
Compd
G-Score
G-energy
LogPo/w
PSA
LogS
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17.
18
19
Std.
5a
5b
5c
5d
5e
5f
5g
5h
5i
À7.33
À7.29
À7.57
À7.41
À7.76
À7.42
À6.91
À6.69
À7.69
À7.18
À6.64
À5.20
À7.20
À7.11
À7.04
À7.34
À7.21
À7.18
À7.03
À5.77
À40.52
À40.47
À58.16
À52.80
À50.96
À51.44
À37.80
À42.66
À40.13
À43.18
À36.21
À38.83
À41.01
À41.06
À41.39
À40.80
À39.54
À34.94
À39.62
À71.55
0.929
1.382
1.274
1.669
1.684
1.405
1.395
1.402
2.218
1.472
1.967
0.944
1.321
1.2
108.199
107.495
116.677
106.747
106.759
107.749
107.691
117.276
106.201
113.399
108.344
105.035
107.229
115.614
105.684
107.042
107.563
105.389
112.844
94.375
À2.40
À2.99
À2.58
À3.29
À3.37
À3.01
À2.98
À2.81
À4.17
À2.91
À3.58
À2.59
À2.88
À2.53
À2.68
À3.05
À3.35
À3.91
À2.38
À4.49
5j
5k
6a
6b
6c
6d
6e
6f
6i
6j
Rosi
1.57
1.513
1.285
2.001
1.181
3.475
The bold values signify the glide score of those compounds are higher than that of
the standard drug Rosiglitazone.
chromones (2a–k) which upon Knoevenagel condensation with
2,4-thiazolidinedione (3) yielded chromonyl-2,4 thiazolidinedi-
ones (4a–k). The catalytic hydrogenation of chromonyl-2,
4-thiazolidinediones (4a–k) using Pd/C in DMF at 30 psi
(H2 gas) for 10–12 h at ambient temperature afforded the
desired products (5a–k) together with compounds (6a–j) as
the by-products as shown in Scheme 1. A library of nineteen
chromone and 2,4-thiazolidinedione conjugates (5a–k) and
(6a–j) have been synthesized in good yields (Supplementary
data).
The synthesized compounds (5a–k) and (6a–j) were docked
for in silico studies against PPAR-
Molecular docking studies were done to gain insights of molecular
binding modes of the ligands with the large pocket of PPAR-
receptors. The compounds were docked against grid generated by
c target (Supplementary data).
c

S. Nazreen et al. / Bioorg. Med. Chem. Lett. 24 (2014) 3034–3042
3037
Schrodinger glide software. In order to analyze the binding pattern
and energies of new molecules & reference ligand (Rosiglitazone),
they were docked individually against the generated grid. Initial
docking of reference ligand (Rosiglitazone) against generated grid
helped in validating the grid generated by software. Rosiglitazone
is reported to have H-bonding with TYR 473, HIS 449 & CYS 285.
Docking of Rosiglitazone against generated grid showed similar
docking mode and H-bonds with RMSD value of 2.8 and hence val-
idated the generated grid. The synthesized molecules docked
against the grid showed good binding energies ranging from
À58.16 to À34.9 kcal/mol. All the molecules showed good glide
score but the most promising molecules were 5c, 5d, 5e, 5f, 5i, 5j
and 6e with best Glide score of À7.57, À7.41, À7.76, À7.42,
À7.69, À7.18 and À7.34, respectively whereas glide score of
Rosiglitazone was À5.77 kcal/mol with respect to same grid. All
the molecules were predicted to bind with large binding site of
bonding with SER 289 residue of the target protein whereas
compounds 5c and 5d were found to be aligned perfectly with the
hydrophobic pocket of the protein. The in silico ADME (Absorption,
Distribution, Metabolism and Excretion) prediction of the
synthesized library were found to be within the acceptable range
(Table 2 and Fig. 1).
In order to validate the molecular docking results, all the syn-
thesized compounds were screened for the in vivo antidiabetic
activity in STZ induced diabetic rat model. The protocol for the
in vitro and in vivo antidiabetic activity has been provided in the
Supplementary data. As observed from the data of Figure 2 among
the synthesized library, five compounds 5c, 5d, 5e, 5f and 5i
showed significant blood glucose lowering effect after 15 days of
study. Compounds 5c and 5e were found to be more potent than
standard drug Rosiglitazone but comparable to another standard
drug Pioglitazone in lowering blood glucose level. Compounds
5d, 5f and 5i also showed significant activity comparable to both
the standards. Compounds 5c, 5d, 5e, 5f, 5i, 5j and 6e with best
PPAR-c receptor with good binding scores with respect to Rosiglit-
azone. Compounds 5e, 5f and 5i were found to form hydrogen
Figure 1. Molecular docking of the active compounds showing interactions with the hydrophobic pocket of the protein. Comparative binding orientation of crystallographic
(green) & docked (purple) structure of Rosiglitazone as docked by Schrodinger software. Hydrogen bonding with Tyr473, His449 & Cys285 is highlighted in red colour.
Compounds 5e, 5i and 5f showing hydrogen bonding with SER 289 (yellow dotted line). Compounds 5c and 5d deeply buried into hydrophobic pocket of PPAR-c.

3038
S. Nazreen et al. / Bioorg. Med. Chem. Lett. 24 (2014) 3034–3042
Fig. 1 (continued)
Glide scores showed significant blood glucose lowering activity.
Compounds 5a, 5b, 6b, 6c, 6d, 6f, 6i and 6j although showing glide
score more than the standard rosiglitazone showed moderate
in vivo activity. Further from oral glucose tolerance test (Fig. 3),
the administration of compound 5e causes significant decrease in
the blood glucose levels of diabetic rats at 120 min when compared
to diabetic control rats indicating that glucose tolerance was
improved by administration of compound 5e. Insulin tolerance test
(Fig. 3) showed that the blood glucose levels of 5e treated diabetic
rats were significantly lowered after 90 min of insulin administra-
tion as compared to diabetic rats indicating insulin resistance was
improved by compound 5e. These results suggest that the antihy-
perglycemic activity of 5e may result from enhanced insulin and
compounds have therefore been assayed for AST, ALT and ALP lev-
els and hepatotoxicity. It has been reported that the levels of these
enzymes are significantly increased in STZ rats indicating the toxic
effect of STZ on liver.24 The elevation of these enzymes might be
due to increased protein catabolism followed by gluconeogenesis
as ALT and AST are directly involved in the amino acid conversion
to ketoacids. It was observed that the levels of serum AST, ALT and
ALP significantly increased in STZ treated rats were significantly
decreased to near normal level after treatment with the active
compounds 5c, 5d, 5e, 5f and 5i as shown in Figure 5. Compounds
5c, 5d, 5e, and 5f were found to be more potent in lowering the
AST, ALT and ALP levels than the standard Pioglitazone whereas
compound 5i was equally potent to Pioglitazone thereby these
compounds have protective effect on liver. Histopathological study
of the liver of the treated animals also showed that two
compounds 5e and 5c did not cause any damage to the liver.
Compound 5d and 5f caused mild damage to the liver that is they
caused mild dilation in sinusoidal space and compound 5i having
disubstituted halogen and standard drug Pioglitazone caused
significant damage to the liver (Fig. 6). TZDs are associated with
cardiovascular risks and are the leading cause for the withdrawal
glucose resistance. As the PPAR-
c agonists are associated with
body weight gain, 5e was also tested for body weight gain study.
Compound 5e treated normal rats did not show any significant
change in body weight as compared to normal control rats. How-
ever, oral administration of 5e to diabetic rats for 15 days caused
a significant improvement in body weight of diabetic rats (Fig. 4).
TZDs are reported to cause hepatotoxicity which is the major
drawback encountered with this class of drug. The synthesized

S. Nazreen et al. / Bioorg. Med. Chem. Lett. 24 (2014) 3034–3042
3039
Figure 2. Antidiabetic activity of the synthesized compounds (5a–k) and (6a–j) in STZ induced diabetic rats. Data is analyzed by one way ANOVA followed by Dunnett’s ‘t’
test and expressed as mean SEM from five observations; ⁄⁄indicates p <0.01 & ⁄indicates p <0.05 versus diabetic control.
Figure 3. Effect of compound 5e on Oral Glucose Tolerance test and Insulin Tolerance test. Data is analyzed by one way ANOVA followed by Dunnett’s ‘t’ test and expressed as
mean SEM from five observations; ⁄indicates p <0.001 versus diabetic control. #indicates p <0.05 versus diabetic control. NC: Normal control; NC + 5e: normal control + 5e;
STZ: diabetic control; STZ + 5e: 5e treated diabetic rats.
of these drugs from the market. We therefore tested compound 5e
for hERG inhibition. It has been reported that compounds with an
reasons. For example, the test compounds 5c–5f and 5i are
administered orally and therefore absorption, metabolism, excre-
tion, etc. of the test compounds might have contributed in exert-
ing the significant lowering in blood glucose level (Reddy et al.
1999).25 Also, compounds 5c–5f and 5i might be exhibiting their
antihyperglycemic activity through other mechanisms, in addition
IC50 >10
cardiotoxicity. Since compound 5e was found to have an IC50 of
135 M, indicating that compound 5e would not be associated
lM do not inhibit hERG significantly and hence have no
l
with cardiotoxicity.
In order to confirm the mode of action of the active com-
to binding to PPAR-c. Similar kind of results have been reported
pounds, the active compounds were tested for in vitro PPAR-
transactivation study. It was observed from the Figure 7 that
these compounds were found to be PPAR- active. Compound
c
by other groups working with TZDs,25 which showed superior
euglycemic and hypolipidemic profiles in db/db mice but not sig-
c
nificant in vitro PPAR-
drug troglitazone.
c transactivation activity than the standard
5e was most potent among the synthesized compounds followed
by 5c and 5d. From these results, it was clear that the compounds
may have exerted the blood glucose lowering effect by activating
From the biological data, the structure activity relationship
(SAR) can be drawn as follows. Compounds (6a–j) having reduced
olefinic bond of chromone ring did not show promising blood glu-
cose lowering effect as compared to compounds (5a–k) wherein
the olefinic bonds remained intact in the chromone ring. Therefore,
olefinic bond of the chromone ring is necessary for the blood
glucose lowering effect. The activity is also dependent on the nat-
ure and position of the substituents on chromone ring as well. Halo
PPAR-
exerted similar or lower blood glucose levels than Pioglitazone
and Rosiglitazone but lower PPAR- transactivation activity than
c receptors. The synthesized compounds 5c–5f and 5i
c
Pioglitazone and Rosiglitazone. The absence of linear correlation
between in vivo pharmacological profile in albino rats and
in vitro PPARc transactivation assays may be attributed to several

3040
S. Nazreen et al. / Bioorg. Med. Chem. Lett. 24 (2014) 3034–3042
Since compound 5e was found to be most PPAR-
was further subjected to PPAR- gene expression study. The
PPAR- gene expression study was done to know the impact of
compound 5e on PPAR- gene. The protocol for the gene expres-
sion study has been provided in the Supplementary data. It was
observed that the PPAR- expression was significantly increased
in presence of compound 5e (2.56-fold) in comparison to stan-
dard drug Pioglitazone (Fig. 8). The increase in PPAR- expression
supports the results of in vitro PPAR- transactivation study. It is
thus clear that the in vitro PPAR- transactivation and in vivo
blood glucose lowering activity of compound may be due to
increase in the PPAR- gene expression. It has reported that TZDs
c active, it
c
c
c
c
c
c
c
c
improve insulin action by effects on gene transcription in the fat
cell that lead to diminished plasma levels of free fatty acids
(FFAs) and an increase in the level of adiponectin, an adipokine
that activates AMPK.26–28 The increase in gene expression by
compound 5e might be due to the activation of AMPK. The over-
expression of PPAR-
c in mature 3T3-L1 adipocytes increases the
Figure 4. Effect of compound 5e on body weight in albino wistar rats. Data is
analyzed by one way ANOVA followed by Dunnett’s ‘t’ test and expressed as
mean SEM from five observations; ⁄indicates p <0.01 versus diabetic control. NC:
Normal control; NC + 5e: normal control + 5e; STZ: diabetic control; STZ + 5e: 5e
treated diabetic rats.
amount of the mRNA for the ubiquitous GLUT1, whose expression
is reported to be downregulated during adipocyte differentia-
tion.29 The reduction of insulin-stimulated glucose transport in
3T3-L1 adipocytes overexpressing PPAR-
c may be due to the
reduced expression of IR, IRS1, IRS2, and GLUT4. Thus, compound
5e increase the gene expression by maintaining insulin sensitivity
in mature 3T3-L1 adipocytes by regulating the expression of
genes that encode components of the insulin signaling pathway
as well as by increasing the expression levels of GLUT1 and
GLUT4 in these cells.
In summary, a library of nineteen compounds have been
synthesized out of which two compounds 5[(6-methoxy-4-
oxo-4H-chromen-3yl) methyl] thia zolidine-2,4-dione (5c) and
5[(6-chloro-4-oxo-4H-chromen-3yl) methyl]thiazolidine-2,4-dione
(5e) exhibited significant in vivo blood glucose lowering effect as
substituted compounds exhibited potent activity whereas meth-
oxy substitution on aromatic ring exhibited good activity. Among
the halogens, the activity was observed in the following order-
Cl > Br > F. Substitution at position 6 of the chromone ring was
more favourable for the activity. Methoxy and halogen substitution
at position 6 exhibited significant blood glucose lowering and
PPAR-
6,8-dichloro substituted compound 5i shows good blood glucose
lowering effect and PPAR- activity but caused significant
c activity than substitution at position 5 and 7. While
c
hepatotoxicity. This may be due to the steric hindrance on the
well as in vitro PPAR-c transactivation which is comparable with
aromatic ring by the two halogens.
the standard drug Pioglitazone and Rosiglitazone. Compounds 5c
Figure 5. Effect of compounds on serum AST, ALT and ALP activities. Values are given as mean SD.

S. Nazreen et al. / Bioorg. Med. Chem. Lett. 24 (2014) 3034–3042
3041
Figure 6. Haematoxylin and eosin immunohistochemical staining of liver after administration of synthesized drugs. Histopathology report of rat liver. As illustrated in above
figure, Low and high power photomicrograph of liver from animal treated groups 5c, 5d, 5e, 5f, 5i and standard 10Â. Low power photomicrograph of liver from corresponding
animal 5c, 5d, 5e, 5f, 5i and standard treated groups showing normal arrangement of cells in the liver lobule. PT = Portal Triad and CV = Central vein 40Â. High power
photomicrograph of liver from animal treated groups 5c, 5d, 5e, 5f, 5i and standard showing normal arrangement of hepatocytes in the centrizonal area. CV = Central Vein.
Compounds 5c and 5e treated groups showing normal arrangement of cells in the liver lobule and normal arrangement of hepatocytes in the centrizonal area. 5d, 5f and
pioglitazone treated groups showing a mild dilatation of sinusoidal spaces around the central vein, 5i treated group showing evident centrizonal sinusoidal dilatation and
thinning of liver cell plates.
Figure 7. PPAR-c transactivation assay of active synthesized compounds. Values are expressed as mean SE from three experiments conducted in triplicate at 10 lM.

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S. Nazreen et al. / Bioorg. Med. Chem. Lett. 24 (2014) 3034–3042
3. Avupati, V. R.; Yejella, R. P.; Akula, A.; Guntuku, G. S.; Doddi, B. R.; Vutla, V. R.;
Anagani, S. R.; Adimulam, L. S.; Vyricharla, A. K. Bioorg. Med. Chem. Lett. 2012,
22, 6442.
4. Cantello, B. C. C.; Cawthorne, M. A.; Cottam, G. P.; Duff, P. T.; Haigh, D.; Hindley,
R. M.; Lister, C. A.; Smith, S. A.; Thurlby, P. L. J. Med. Chem. 1994, 37, 3977.
5. Prabhakar, C.; Madhusudhan, G.; Sahadev, K.; Maheedhara Reddy, C.; Sarma, M.
R.; Om Reddy, G.; Chakrabarti, R.; Seshagiri Rao, C.; Dileep Kumar, T.;
Rajagopalan, R. Bioorg. Med. Chem. Lett. 1998, 8, 2725.
6. Sunduru, N.; Srivastava, K.; Rajakumar, S.; Puri, S. K.; Saxena, J. K.; Chauhan, P.
M. S. Bioorg. Med. Chem. Lett. 2009, 19, 2570.
7. Reddy, K. A.; Lohray, B. B.; Bhushan, V.; Reddy, A. S.; Kishore, P. H.; Rao, V. V.;
Saibaba, V.; Bajji, A. C.; Rajesh, B. M.; Reddy, K. V.; Chakrabarti, R.; Rajagopalan,
R. Bioorg. Med. Chem. Lett. 1998, 8, 999.
8. Hafez, H. N.; ElGazzar, A. R. B. A. Bioorg. Med. Chem. Lett. 2009, 19, 4143.
9. Patil, V.; Tilekar, K.; Munj, S. M.; Mohan, R.; Ramaa, C. S. Eur. J. Med. Chem. 2010,
45, 4539.
10. Bonde, C. G.; Gaikwad, N. J. Bioorg. Med. Chem. 2004, 12, 2151.
11. Fan, Y. H.; Chen, H.; Natarajan, A.; Guo, Y.; Harbinski, F.; Iyasere, J.; Christ, W.;
Aktasa, H.; Halperina, J. A. Bioorg. Med. Chem. Lett. 2004, 14, 2547.
12. Jeon, R.; Park, S. Y. Arch. Pharm. Res. 2004, 27, 1099.
Figure 8. Effect of compound 5e on PPAR-c gene expression.
13. Rikimaru, K.; Wakabayashi, T.; Abe, H.; Imoto, H.; Maekawa, T.; Ujikawa, O.;
Murase, K.; Matsuo, T.; Matsumoto, M.; Nomura, C.; Tsuge, H.; Arimura, N.;
Kawakami, K.; Sakamoto, J.; Funami, M.; Mol, C. D.; Snell, G. P.; Bragstad, K. A.;
Sang, B. C.; Dougan, D. R.; Tanaka, T.; Katayama, N.; Horiguchi, Y.; Momose, Y.
Bioorg. Med. Chem. 2012, 20, 714.
14. Willson, T. M.; Lambert, M. H.; Kliewer, S. A. Annu. Rev. Biochem. 2001, 70, 341.
15. Hong, W. L.; Joong, B. A.; Sung, K. K.; Soon, K. A.; Deok, C. H. Org. Process Res.
Dev. 2007, 11, 190.
16. Packiavathy, A. S. C.; Ramalingam, M.; Devi, C. A. Orient. J. Chem. 2013, 3, 7.
17. Jawale, D. V.; Pratap, U. R.; Mane, R. A. Bioorg. Med. Chem. Lett. 2012, 22, 924.
18. Iqbal, A. K. M.; Khan, A. Y.; Kalashetti, M. B.; Belavagi, N. S.; Gong, Y. D.; Khazi, I.
A. M. Eur. J. Med. Chem. 2012, 53, 308.
and 5e recovered the activity of serum AST, ALT and ALP level and
did not cause any damage to the liver. Compound 5e has significant
effect on PPAR-c gene expression as it increases the PPAR-c expres-
sion by 2.56-fold in comparison to Pioglitazone. Compounds 5c and
5e may be considered as promising candidate for development of
new antidiabetic agents.
Acknowledgements
19. Nesto, R. W.; Bell, D.; Bonow, R. O.; Fonseca, V.; Grundy, S. M.; Horton, E. S.;
Winter, M. L.; Porte, D.; Semenkovich, C. F.; Smith, S.; Young, L. H.; Kahn, R.
Diabetes Care 2004, 27, 256.
20. Hollman, P. C.; Katan, M. B. Food Chem. Toxicol. 1999, 37, 937.
21. Beecher, G. R. J. Nutr. 2003, 133, 3248S.
22. Moon, Y. J.; Wang, X.; Morris, M. E. Toxicology 2006, 20, 187.
23. Xie, W.; Wang, W.; Su, H.; Xing, D.; Pan, Y.; Du, L. Comp. Biochem. Physiol. C:
Toxicol. Pharmacol. 2006, 143, 429.
The author thanks Vice Chancellor, Dr. G. N. Qazi for providing
necessary facilities to the Department of Chemistry. Syed Nazreen
acknowledges University Grants Commission, New Delhi, for finan-
cial assistance.
24. Shinde, U. A.; Goyal, R. K. J. Cell. Mol. Med. 2003, 7, 332.
Supplementary data
25. Reddy, K. A.; Lohray, B. B.; Bhushan, V.; Reddy, A. S.; Rao Mamidi, N. V.; Reddy,
P. P.; Saibaba, V.; Reddy, N. J.; Suryaprakash, A.; Misra, P.; Vikramadithyan, R.
K.; Rajagopalan, R. J. Med. Chem. 1999, 42, 3265.
26. Yamauchi, T.; Kamon, J.; Waki, H.; Murakami, K.; Motojima, K.; Komeda, K.; Ide,
T.; Kubota, N.; Terauchi, Y.; Tobe, K.; Miki, H.; Tsuchida, A.; Akanuma, Y.; Nagai,
R.; Kimura, S.; Kadowaki, T. J. Biol. Chem. 2001, 276, 41245.
Supplementary data associated with this article can be found,
in the online version, at http://dx.doi.org/10.1016/j.bmcl.2014.
05.034.
27. Maeda, N.; Takahashi, M.; Funahashi, T.; Kihara, S.; Nishizawa, H.; Kishida, K.;
Nagaretani, H.; Matsuda, M.; Komuro, R.; Ouchi, N.; Kuriyama, H.; Hotta, K.;
Nakamura, T.; Shimomura, I.; Matsuzawa, Y. Diabetes 2001, 50, 2094.
28. Nathan, K. L.; Kelly, M.; 1 Tsao, T.; Farmer, S. R.; Saha, A. K.; Ruderman, N. B.;
Tomas, E. Am. J. Physiol. Endocrinol. Metab. 2006, 291, E175.
References and notes
1. Rotella, D. P. J. Med. Chem. 2004, 47, 4111.
29. Wu, Z.; Rosen, E. D.; Brun, R.; Hauser, S.; Adelmant, G.; Troy, A. E.; McKeon, C.;
Darlington, G. J.; Spiegelman, B. M. Mol. Cell 1999, 3, 151.
2. Liu, K.; Xu, L.; Szalkowski, D.; Li, Z.; Ding, V.; Kwei, G.; Huskey, S.; Moller, D. E.;
Heck, J. V.; Zhang, B. B.; Jones, A. B. J. Med. Chem. 2000, 43, 3487.