Synthesis, characterization, and biological evaluation of thiazolidine-2,4-dione derivatives

Article abstract of DOI:10.1007/s00044-013-0705-2

As a part of our continuation studies in developing new derivatives as dual antimicrobial/antitumor agents we describe the synthesis of new (Z)-2-(5-arylidene-2,4-dioxothiazolidin-3-yl) acetic acid derivatives (3a-m). The chemical structures of the compound were elucidated by FTIR, ¹H NMR, ¹³C NMR, and elemental analysis data. The antimicrobial activity of all products was examined. All newly synthesized compounds were tested for their in vitro anticancer activity against four cancer cell lines. Among the synthesized compounds, 3a exhibited notable activity against HeLa, HT29, A549, and MCF-7 cell lines with IC₅₀ values of 55, 40, 38, and 50 μM, respectively. In order to predict the drug likeliness of the synthesized compounds on the guidelines of Lipinski rule of five studies was carried out using Pallas software.

Full text of DOI:10.1007/s00044-013-0705-2

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res (2014) 23:987–994
DOI 10.1007/s00044-013-0705-2
ORIGINAL RESEARCH
Synthesis, characterization, and biological evaluation
of thiazolidine-2,4-dione derivatives
•
Shankar G. Alegaon Kallanagouda R. Alagawadi
•
•
Sneha M. Pawar D. Vinod Udaysingh Rajput
•
Received: 26 April 2013 / Accepted: 8 August 2013 / Published online: 21 August 2013
Ó Springer Science+Business Media New York 2013
Abstract As a part of our continuation studies in devel-
oping new derivatives as dual antimicrobial/antitumor
agents we describe the synthesis of new (Z)-2-(5-arylidene-
2,4-dioxothiazolidin-3-yl) acetic acid derivatives (3a–m).
The chemical structures of the compound were elucidated
by FTIR, 1H NMR, 13C NMR, and elemental analysis data.
The antimicrobial activity of all products was examined.
All newly synthesized compounds were tested for their
in vitro anticancer activity against four cancer cell lines.
Among the synthesized compounds, 3a exhibited notable
activity against HeLa, HT29, A549, and MCF-7 cell lines
with IC50 values of 55, 40, 38, and 50 lM, respectively. In
order to predict the drug likeliness of the synthesized
compounds on the guidelines of Lipinski rule of five
studies was carried out using Pallas software.
Introduction
The development of novel antimicrobial and anticancer
therapeutic agents is one of the fundamental goals in
medicinal chemistry. Despite major breakthroughs in many
areas of modern medicine, significant rise in multi-drug
resistant microbial infections and cancer has become an
economic as well as a serious health care problem. In some
microbial infections, strains of microbes have become
resistant to all available drugs. There is an increasing need
for new medicinal organic agent because a dose of anticancer
drug sufficient to kill cancer cells is often lethal to the normal
tissue and leads to many side effects, which in turn, limits its
treatment efficacy (Aydemir and Bilaloglu, 2003). The thi-
azolidine-2,4-dione (TZDs) is an imperative scaffold that is
not only synthetically important but also possesses a wide
range of promising biological activities. The TZDs moiety is
extensively utilized as a carboxylic acid mimetic to improve
the metabolic stability and therapeutic profile of bioactive
agents (Boyd, 2007; McIntyre et al., 2007; Elte and Blickle,
2007). During recent years there has been a large investi-
gation on different classes of TZDs compounds, many of
these are known to possess antibacterial and antifungal
(Ayhan-Kilcigil and Altanlar, 2000; Heerding et al., 2003;
Bozdag-Dundar et al., 2007; Tuncbilek and Altanlar, 2006)
properties. In addition, this class of compounds has several
other potentially beneficial effects including on lipid profile,
blood pressure lowering, and anti-inflammatory effects
(Kalaitzidis et al., 2009). TZDs target vascular cells (Kure-
bayashi et al., 2005) and monocytes/macrophages (Jiang
et al., 1998; Ricote et al., 1998) to inhibit the production of
pro-inflammatory cytokines as well as the expression of
inducible nitric oxide synthase and cell adhesion molecules.
These drugs may also be beneficial in multiple sclerosis and
neurodegenerative diseases, including Alzheimer’s and
Keywords Antimicrobial activity Á Cytotoxic activity Á
Thiazolidine-2,4-dione Á Drug likeliness
Electronic supplementary material The online version of this
article (doi:10.1007/s00044-013-0705-2) contains supplementary
material, which is available to authorized users.
S. G. Alegaon (&) Á K. R. Alagawadi Á S. M. Pawar
Department of Pharmaceutical Chemistry, KLE University’s
College of Pharmacy, Nehrunagar, Belgaum 590010, Karnataka,
India
e-mail: sgalegaon@gmail.com; sgalegaon@klepharm.edu
D. Vinod
Department of Pharmaceutical Chemistry, College of Pharmacy,
Madras Medical College, Chennai, Tamil Nadu, India
U. Rajput
Department of Chemistry, R. L. Science Institute, College Road,
Belgaum 590001, Karnataka, India
123

988
Med Chem Res (2014) 23:987–994
Parkinson’s, at least partially due to their anti-inflammatory
activity (Heneka and Landreth, 2007). These observations
have encouraged us to synthesize some new compounds with
potential biological activity. Hence, herein, we report the
synthesis, characterization, and investigation of antimicro-
bial and cytotoxic properties of (Z)-2-(5-benzylidene-2,4-
dioxothiazolidin-3yl)acetic acid derivatives.
the 5-methylidene carbon and C-5 of the thiazolidinedione
ring resonated at d 113.46 and 128.02 ppm, respectively. In
the 1H NMR spectra of compounds (3a–m), a signal
attributable to N–CH2 resonance was diagnostic: in partic-
ular 1H NMR spectra showed a singlet at d 4.61–4.74 ppm
for acetic acid derivatives. In 13C NMR spectra, besides two
signals attributable to the resonances of 2- and 4-carbonylic
groups of the TZDs ring at 160.31–169.16 ppm, another
signal due to the resonances of the carboxylic carbon
appeared in the same range.
Results and discussion
Chemistry
Antimicrobial activity
The synthesis of (Z)-2-(5-arylidene-2,4-dioxothiazolidin-3-
yl)acetic acids (3a–m) was achieved through the versatile
and efficient synthetic route outlined in Scheme 1. The
starting material TZDs (1) was prepared according to ear-
lier reported method (Prashanth Kumar et al., 2006) with
minor modification. The condensation of compound (1)
with methylbromoacetate in the presence of potassium
carbonate as a base, followed by acidic hydrolysis pro-
vided compound (2,4-dioxothiazolidin-3-yl)acetic acid (2)
(Maccari et al., 2005). Further, the condensation of com-
pound (2) with the appropriate substituted aldehydes, using
piperidine as base in refluxing ethanol for 24 h, provided
compounds (3a–m). Structure of the synthesized com-
pounds was established on the basis of physicochemical,
elemental analysis, and spectral data. The (Z)-4-(5-arylid-
ene-2,4-dioxothizolidin-3-yl) methyl benzoic acid deriva-
tives were prepared and characterized previously (Maccari
et al., 2005; Bruno et al., 2002; Rakowitz et al., 2006). The
title compounds (Z)-4-(5-arylidene-2,4-dioxothizolidin-3-
yl) methyl benzoic acids (3a–m) were obtained only as
Z configuration, as expected from the literature and con-
The newly synthesized compounds were evaluated for their
in vitro antibacterial activity against two Gram-positive bac-
teria namely, Staphylococcus aureus (ATCC 25923) and
Enterococcus faecalis (ATCC 35550); two Gram-negative
bacteria namely, Escherichia coli (ATCC 35218) and Pseu-
domonas aeruginosa (ATCC 25619); and four fungal strains
of Candida albicans (ATCC 2091), Aspergillus flavus (NCIM
No. 524), Aspergillus niger (ATCC 6275), and Cryptococcus
neoformans (Clinical isolate) fungi. The MIC values were
determined by using the twofold serial dilution technique in
Mueller–Hinton broth and Sabouraud dextrose agar for the
antibacterial and antifungal assays, respectively (National
Committee for Clinical Laboratory Standards, 2000). Cipro-
floxacin was used as the reference standard for antibacterial
activitywhileketoconazolewasusedasthereferencestandard
for antifungal activity. Minimum Inhibitory Concentration
(MIC) was recorded as the lowest concentration of a com-
pound that inhibits the growth of tested microorganisms.
The MIC results of the in vitro antibacterial and antifungal
activity screening of a novel series of (Z)-(5-arylidene-2,4-
dioxothiazolidin-3-yl)acetic acids (3a–m) are summarized
in Table 1. All the reported compounds exhibited moderate
to substantial in vitro anti-microbial activity against the
tested bacterial strains. At the outset it can be stated that the
series of (Z)-(5-arylidene-2,4-dioxothiazolidin-3-yl)acetic
acids have displayed more promising and interesting anti-
bacterial and antifungal activities. Among the compounds
1
firmed by H NMR, 13C NMR spectra, and X-ray crystal-
lographic data (Tan et al., 1986; Albuquerque et al., 1995;
1
Vogeli et al., 1978; Bruno et al., 2002). The H NMR
spectra of the title compounds (3a–m) showed only one
signal attributable to the resonance of the 5-methylidene
proton in the range d 7.50–8.03 ppm. In 13C NMR spectra,
Scheme 1 Preparation of 2-(5-
arylidene-2,4-dioxothiazolidin-
O
O
O
O
H
N
H
O
Br
S
Cl
O
O
O
OH
N
3yl)acetic acid derivatives.
Reagents and conditions:
(a) (I) H2O, RT 3 h, (II) HCl,
110 °C for 15 h, (b) (I) K2CO3,
acetone, reflux, 24 h, (II)
AcOH, HCl, reflux, 2 h
H
H
S
N
H
N
H
S
(b)
(a)
O
(1)
(2)
(c) Piperidine, acetic acid,
ethanol reflux, 24 h
H
O
O
O
O
O
OH
OH
Ar
(c)
N
N
+
Ar
H
S
S
O
O
(2)
(3a-m)
123

Med Chem Res (2014) 23:987–994
989
Table 1 In vitro antimicrobial activity of (Z)-2-(5-arylidene-2,4-dioxothiazolidin-3yl)acetic acid derivatives 3a–m expressed as Minimum
inhibitory concentration (MIC) in lg/ml
Compounds
Ar
E.c
P.a
S.a
E.f
C.a
C.n
A.f
A.n
3a
2-ClC6H4
[128
[128
[128
[128
[128
[128
[128
[128
[128
[128
64
[128
64
64
16
64
64
64
64
64
32
16
[128
16
16
64
2
32
16
64
64
64
64
32
32
32
[128
16
16
16
2
64
16
16
32
16
64
3b
2,3-di-Cl C6H3
2-Cl-5-NO2 C6H3
4-Cl-3-NO2 C6H3
2,3-di-OH C6H3
2-OH-5-NO2 C6H3
3,5-di-Br-4-OH C6H2
4-Br-2-F C6H3
4-Br-3-F C6H3
2,4-di-NO2 C6H3
2-Furyl
32
16
3c
64
128
[128
[128
64
128
[128
64
16
16
3d
128
[128
[128
[128
64
128
64
128
[128
[128
32
3e
3f
[128
64
[128
32
3g
32
3h
32
16
32
32
3i
64
16
16
32
16
3j
[128
32
64
[128
64
[128
32
[128
16
3k
16
3l
2-Thienyl
32
16
16
16
32
32
3m
Cipro.
Keto.
1-Methylpyrrol
64
32
16
32
32
16
2
2
NT
2
NT
1
NT
2
NT
1
NT
NT
NT
NT
NT not tested, E.c Escherichia coli, P.a Pseudomonas aeruginosa, S.a Staphylococcus aureus, E.f Enterococcus faecalis, C.a Candida albicans,
C.n Cryptococcus neoformans, A.f Aspergillus flavus, A.n Aspergillus niger, Cipro Ciprofloxacin, Keto Ketoconazole
tested, compounds 3b, 3k, and 3l exhibited excellent anti-
bacterial activity against Gram-positive bacteria like
S. aureus and E. faecalis with a MIC value of 16 lg/ml.
Compounds 3i, 3l, and 3m exhibited significant inhibitory
activity against S. aureus, P. aeruginosa, and E. faecalis with
a MIC value of 16 lg/ml, respectively. However, all other
compounds in the series were found to have least or poor
activity against both Gram-positive and Gram-negative
bacteria as compared to standard. From the antifungal data
through MIC studies, it is evident that most of the tested
compounds exhibited excellent antifungal activity, com-
pounds 3b, 3i, 3k, 3l, and 3m displayed good antifungal
activity against C. albicans, and compounds 3b and 3c
demonstrated significant inhibitory activity against A. flavus,
A. niger with a MIC value of 16 lg/ml, also compounds 3i,
3k, and 3m showed good activity against A. niger with a MIC
value of 16 lg/ml.
(cervical carcinoma), HT29 (colorectal cancer), A549
(lung cancer), MCF-7 (breast adenocarcinoma) cell lines
(Mosmann, 1983). The inhibition of the cell proliferation
was determined 24 h after cells were exposed to the tested
compounds. The IC50 (the concentration that causes 50 %
growth inhibition) values were determined and summa-
rized in Table 2. Results showed that most of the com-
pounds exhibited a considerable effect against A549 (lung
cancer) and MCF-7 (breast adenocarcinoma) cell lines.
Compounds 3a, 3b, 3g, 3k, and 3l showed good cytotox-
icity against A549 with IC50 values of 38, 37, 45, 36, and
30 lM, respectively, Compounds 3a, 3c, 3e, 3f, 3g, 3j, 3l,
and 3m displayed significant inhibitory activity with an
IC50 values ranging from 40–50 lM. In this work we also
performed a computational study for prediction of physi-
cal–chemical properties of all derivatives and the results
are summarized in Table 3. In an effort to study the
hydrophobic pattern, we analyzed the theoretical parame-
ters related to the oral bioavailability according to Lipinski
‘‘rule of 5.’’ This rule theoretically determines if a chemical
compound presents absorption and permeation across the
membranes properties that would make it a likely orally
active drug in humans. Interestingly our results pointed that
most of the compounds obey the Lipinski ‘‘rule of 5.’’ We
Cytotoxic activity
The MTT [3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyl-tetra-
zolium bromide] cell proliferation assay was used to
evaluate cytotoxic activity of the synthesized compounds
against four human cancer cell lines including HeLa
123

990
Med Chem Res (2014) 23:987–994
Table 2 In vitro cytotoxicity profile of (Z)-2-(5-arylidene-2,4-dioxothiazolidin-3yl)acetic acid derivatives 3a–m against selected human cancer
cell lines IC50 (lM)
Compound
Ar
HeLa
HT29
A549
MCF7
3a
2-ClC6H4
55
65
40
57
38
37
65
70
50
54
45
60
65
60
36
30
60
2
50
55
48
60
50
50
50
70
80
40
54
40
40
3
3b
2,3-di-Cl C6H3
2-Cl-5-NO2 C6H3
4-Cl-3-NO2 C6H3
2,3-di-OH C6H3
2-OH-5-NO2 C6H3
3,5-di-Br-4-OH C6H2
4-Br-2-F C6H3
4-Br-3-F C6H3
2,4-di-NO2 C6H3
2-Furyl
3c
77
82
3d
59
70
3e
[100
94
[100
100
79
3f
3g
75
3h
62
83
3i
79
81
3j
65
76
3k
50
40
3l
2-Thienyl
60
50
3m
Cisplatin
1-Methylpyrrol
–
50
60
7
4
Cell lines include cervical carcinoma (HeLa); colorectal cancer (HT29); lung cancer (A549); breast adenocarcinoma (MCF-7)
Table 3 Drug likeliness of the (Z)-2-(5-arylidene-2,4-dioxothiazolidin-3yl)acetic acid derivatives 3a–m
Compound
Ar
MW
Log P
HBD
HBA
Rule-5 violation
3a
3b
3c
3d
3e
3f
2-ClC6H4
297
332
342
342
295
324
437
360
360
353
253
269
266
1.42
2.05
1.36
1.36
0.03
0.67
2.01
1.69
1.69
0.64
0.05
0.69
0.18
1
1
1
1
3
2
2
1
1
1
1
1
1
5
5
0
0
0
0
0
0
0
0
0
1
0
0
0
2,3-di-Cl C6H3
2-Cl-5-NO2 C6H3
4-Cl-3-NO2 C6H3
2,3-di-OH C6H3
2-OH-5-NO2 C6H3
3,5-di-Br-4-OH C6H2
4-Br-2-F C6H3
4-Br-3-F C6H3
2,4-di-NO2 C6H3
2-Furyl
8
8
7
9
3g
3h
3i
6
5
5
3j
11
6
3k
3l
2-Thienyl
6
3m
1-Methylpyrrol
5
MW molecular weight, Log P calculated using software, HBD hydrogen bond donors, HBA hydrogen bond acceptor; Lipinski rule of 5:
(Molecular weight B500, Log P B5, HBD B5 and HBA B10) study was carried out using Pallas and molinspiration software
123

Med Chem Res (2014) 23:987–994
991
used Molinspiration (Ertl et al., 2000) and Pallas (Pallas,
2010) software for calculations of physical–chemical
properties.
to provide (2,4-dioxothiazolidin-3-yl)acetic acid methyl
ester as oil; glacial acetic acid (17 ml) and 2 N HCl
(4.2 ml) were refluxed for 2 h. After evaporation to dryness
in vacuo, the crude oil was washed with water and then
ethanol to provide pure (2,4-dioxothiazolidin-3-yl)acetic
acid as oil (2). Piperidine and suitable substituted alde-
hydes were added to a solution of (2,4-dioxothiazolidin-3-
yl)acetic acid in ethanol and the mixture was refluxed for
24 h. The reaction mixture was then poured into water and
acidified with acetic acid to give as crude solid (3a–m),
which was recrystallized from appropriate solvents.
Conclusion
In conclusion, we report the synthesis of (Z)-(5-arylidene-
2,4-dioxothiazolidin-3-yl) acetic acid derivatives and
evaluated for their in vitro antimicrobial and cytotoxic
activities. The biological activities of (Z)-(5-arylidene-2,4-
dioxothiazolidin-3-yl) acetic acid depended upon the sub-
stituent, compound 3a exhibited notable in vitro anticancer
activity against HeLa, HT29, A549, and MCF-7 cell lines
with IC50 values of 55, 40, 38, and 50 lM, respectively.
Moreover, most of the synthesized compounds obey the
Lipinski ‘‘rule of 5.’’ Thus the compounds containing 2,4-
(dioxothiazolidin-3-yl)acetic acid could constitute an
interesting lead for the evaluation of antimicrobial and
cytotoxic activity and may be helpful for the design of new
therapeutic agents or for further lead optimization.
(Z)-2-(5-(2-chlorobenzylidene)-2,4-dioxothizolidin-3-
yl)acetic acid (3a) This compound was prepared
according to general procedure and it was obtained as a
yellow solid, mp 215–217 °C. IR (KBr, cm-1) 3364 (OH
of COOH), 1710 (C=O), 1680 (C=O). 1H NMR (400 MHz,
d, ppm, DMSO-d6) 7.95 (s, 1H, –CH=), 7.52–7.68 (m, 4H,
Ar–H), 4.72 (s, 2H, CH2). 13C NMR (400 MHz, d, ppm,
DMSO-d6) 167.06 (C=O, TZD), 165.19 (C=O, –CH2C=O),
164.33 (C=O, TZD), 147.38, 141.20, 137.77, 133.75,
130.24, 119.66, 114 (ArC), 43.31(CH2). Elemental Anal.
Calcd for (C12H8ClNO4S) C, 48.41; H, 2.71; N, 4.70.
Found: C, 48.40; H, 2.69; N, 4.73.
Experimental protocols
General conditions
(Z)-2-(5-(2,3-dichlorobenzylidene)-2,4-dioxothizolidin-3-
yl)acetic acid (3b) This compound was prepared
according to general procedure and it was obtained as a
yellow solid, mp 210–213 °C. IR (KBr, cm-1) 3385 (OH
of COOH), 1705 (C=O), 1683 (C=O). 1H NMR (400 MHz,
d, ppm, DMSO-d6) 7.94 (s, 1H, –CH=), 7.80 (d, 1H, Ar–
H). 7.51–7.60 (m, 2H, Ar–H), 4.74 (s, 2H, CH2). 13C NMR
(400 MHz, d, ppm, DMSO-d6) 167.36 (C=O, TZD),
166.90 (C=O, –CH2C=O), 165.50 (C=O, TZD), 141.11,
140.30, 133.95, 127.50, 125.53, 123.95, 113.46 (ArC),
44.46 (CH2). Elemental Anal. Calcd for (C12H7Cl2NO4S)
C, 43,39; H, 2.21; N, 4.22. Found: C, 43.36; H, 2.09; N,
4.26.
All the chemicals used in this study were purchased from
Aldrich, Himedia, and SD fine chemicals. Melting points
were determined in open capillary tubes and are uncor-
rected. The progress of the reactions was monitored by
TLC F 254 Merck precoated silica gel plates. The IR
spectra were recorded on Nicolet Impact 410 FTIR spec-
1
trophotometer using KBr pellets. H NMR spectra were
recorded on AMX-400, Bruker-400 liquid-state NMR
spectrometer using tetramethylsilane (TMS) as the internal
standard. Chemical shifts were recorded as d (ppm). Ele-
mental analysis was carried out using a Perkin Elmer
2400-CHN Analyzer. Microwave (MW) experiments were
carried out in a domestic oven operating at a frequency of
2450 MHz. Spectra facilities were carried out by Sophis-
ticated Analytical Instruments Facility (SAIF) division of
Indian Institute of Science, Bangalore, India.
(Z)-2-(5-(2-chloro-5-nitrobenzylidene)-2,4-dioxothizolidin-
3-yl)acetic acid (3c) This compound was prepared
according to general procedure and it was obtained as a
yellow solid, mp 221–222 °C. IR (KBr, cm-1) 3360 (OH
of COOH), 1705 (C=O), 1680 (C=O). 1H NMR (400 MHz,
d, ppm, DMSO-d6) 8.31–8.34 (m, 1H, Ar–H), 7.95–7.97
Chemistry
(m, 2H, Ar–H), 7.89 (s, 1H, –CH=), 4.69 (s, 2H, CH2). 13
C
General procedure for the preparation of (Z)-2-(5-
NMR (400 MHz, d, ppm, DMSO-d6) 167.09 (C=O, TZD),
165.23 (C=O, –CH2C=O), 164.28 (C=O, TZD), 138.43,
137.65, 133.01, 129.12, 128.02, 125.56, 119.18, 107.74
(ArC), 43.29 (CH2). Elemental Anal. Calcd for
(C12H7ClN2O6S) C, 42.06; H, 2.06; N, 8.17. Found: C,
42.02; H, 2.02; N, 8.19.
arylidene-2,4-dioxothiazolidin-3-yl)acetic acids (3a–m)
A mixture of TZDs 1 (10 mmol), methylbromoacetate
(20 mmol), and potassium carbonate (20 mmol) was
refluxed for 24 h; then the solvent was evaporated under
reduced pressure. The residue was washed with methanol
123

992
Med Chem Res (2014) 23:987–994
(Z)-2-(5-(4-chloro-3-nitrobenzylidene)-2,4-dioxothizolidin-
3-yl)acetic acid (3d) This compound was prepared
according to general procedure and it was obtained as a
yellow solid, mp 230–231 °C. IR (KBr, cm-1) 3374 (OH
of COOH), 1715 (C=O), 1685 (C=O). 1H NMR (400 MHz,
d, ppm, DMSO-d6) 8.39 (d, 2H, Ar–H), 7.93 (s, 1H,
–CH=), 7.89–7.92(m, 1H, Ar–H), 4.67 (s, 2H, CH2). 13C
NMR (400 MHz, d, ppm, DMSO-d6). 167.01 (C=O, TZD),
165.08 (C=O, –CH2C=O), 164.33 (C=O, TZD), 139.75,
137.79, 133.93, 128.27, 119.84, 107.74 (ArC), 43.31
(CH2). Elemental Anal. Calcd for (C12H7ClN2O6S) C,
42.06; H, 2.06; N, 8.17. Found: C, 42.03; H, 2.02; N, 8.19.
(Z)-2-(5-(4-bromo-2-fluorobenzylidene)-2,4-dioxothizolidin-
3-yl)acetic acid (3h) This compound was prepared
according to general procedure and it was obtained as a
orange solid, mp 214–216 °C. IR (KBr, cm-1) 3365 (OH
of COOH), 1705 (C=O), 1680 (C=O). 1H NMR (400 MHz,
d, ppm, DMSO-d6) 7.75–7.78 (dd, 1H, Ar–H), 7.50 (s, 1H,
–CH=), 7.59–7.61 (dd, 1H, Ar–H), 7.49–7.53 (t, 1H, Ar–
H), 4.66 (s, 2H, CH2). 13C NMR (400 MHz, d, ppm,
DMSO-d6) 169.16 (C=O, TZD), 166.12 (C=O, –CH2C=O),
162.32 (C=O, TZD), 147.60, 130.66, 128.43, 123.67,
121.07, 119.08, 119.69 (ArC), 39.87 (CH2). Elemental
Anal. Calcd for (C12H7BrFNO4S) C, 40.02; H, 1.96; N,
3.89. Found: C, 40.01; H, 1.92; N, 3.88.
(Z)-2-(5-(2-3-dihydroxybenzylidene)-2,4-dioxothizolidin-3-
yl)acetic acid (3e) This compound was prepared
according to general procedure and it was obtained as a
yellow solid, mp 212–213 °C. IR (KBr, cm-1) 3360 (OH
of COOH), 1710 (C=O), 1685 (C=O). 1H NMR (400 MHz,
d, ppm, DMSO-d6) 9.84 (s, 1H, Ar–OH), 9.65 (s, 1H, Ar–
OH), 8.03 (s, 1H, –CH=), 6.91–6.93 (m, 1H, Ar–H),
6.75–6.84 (m, 2H, Ar–H), 4.61 (s, 2H, CH2). 13C NMR
(400 MHz, d, ppm, DMSO-d6) 166.02 (C=O, TZD),
165.41 (C=O, –CH2C=O), 160.49 (C=O, TZD), 142.21,
141.46, 126.53, 122.72, 121.31, 118.59, 117.65 (ArC),
41.69 (CH2). Elemental Anal. Calcd for (C12H9NO6S) C,
48.81; H, 3.07; N, 4.74. Found: C, 48.78; H, 3.05; N, 4.72.
(Z)-2-(5-(4-bromo-3-fluorobenzylidene)-2,4-dioxothizolidin-
3-yl)acetic acid (3i) This compound was prepared
according to general procedure and it was obtained as
orange solid, mp 224–226 °C. IR (KBr, cm-1) 3370 (OH
of COOH), 1704 (C=O), 1685 (C=O). 1H NMR (400 MHz,
d, ppm, DMSO-d6) 8.05–8.07 (dd, 1H, Ar–H), 7.86 (s, 1H,
–CH=), 7.67–7.70 (m, 1H, Ar–H), 7.54–7.58 (t, 1H, Ar–H),
4.62 (s, 2H, CH2). 13C NMR (400 MHz, d, ppm, DMSO-
d6) 165.11 (C=O, TZD), 164.11 (C=O, –CH2C=O),
160.31(C=O, TZD), 141.08, 134.54, 130.16, 129.09,
128.48, 125.61, 116.04 (ArC), 42.09 (CH2). Elemental
Anal. Calcd for (C12H7BrFNO4S) C, 40.02; H, 1.96; N,
3.89. Found: C, 40.00; H, 1.95; N, 3.88.
(Z)-2-(5-(2-hydroxy-5-nitrobenzylidene)-2,4-dioxothizolidin-
3-yl)acetic acid (3f) This compound was prepared accord-
ing to general procedure and it was obtained as a yellow solid,
mp 205–206 °C. IR (KBr, cm-1) 3370 (OH of COOH), 1705
(C=O) 1680 (C=O). 1H NMR (400 MHz, d, ppm, DMSO-d6)
8.22 (d, 1H, Ar–H), 8.11 (d, 1H, Ar–H), 7.87 (s, 1H, –CH=),
6.87(d, 1H, Ar–H), 4.72(s, 2H, CH2). 13C NMR (400 MHz, d,
ppm, DMSO-d6) 167.11 (C=O, TZD), 165.52 (C=O,
–CH2C=O), 164.13 (C=O, TZD), 138.06, 137.16, 133.31,
129.16, 128.14, 124.56, 119.32, 108.48 (ArC), 43.31(CH2).
Elemental Anal. Calcd for (C12H8N2O7S) C, 44.45; H, 2.49;
N, 8.64. Found: C, 44.40; H, 2.46; N, 8.69.
(Z)-2-(5-(2,4-dinitrobenzylidene)-2,4-dioxothizolidin-3-yl)-
acetic acid (3j) This compound was prepared according to
general procedure and it was obtained as yellow solid, mp
209–210 °C. IR (KBr, cm-1) 3350 (OH of COOH), 1700
(C=O), 1680 (C=O). 1H NMR (400 MHz, d, ppm, DMSO-
d6) 8.16 (s, 1H, –CH=), 8.13–8.14 (dd, 1H, Ar–H), 7.78–7.79
(dd, 1H, Ar–H), 7.31–7.34(m, 1H, Ar–H), 4.68 (s, 2H, CH2).
13C NMR (400 MHz, d, ppm, DMSO-d6) 166.95 (C=O,
TZD), 165.09 (C=O, –CH2C=O), 165.23 (C=O, TZD),
138.34, 131.95, 129.12, 124.32, 109.67 (ArC), 41.09 (CH2).
Elemental Anal. Calcd for (C12H7N3O8S) C, 40.80; H, 2.00;
N, 11.89. Found: C, 40.75; H, 1.98; N, 11.93.
(Z)-2-(5-(3,5-dibromo-4-hydroxybenzylidene)-2,4-dioxo-
thizolidin-3-yl)acetic acid (3g) This compound was pre-
pared according to general procedure and it was obtained
as a yellow solid, mp 231–233 °C. IR (KBr, cm-1) 3355
(OH of COOH), 1710 (C=O) 1690 (C=O). 1H NMR
(400 MHz, d, ppm, DMSO-d6) 8.47 (s, 1H, Ar–OH), 7.74
(s, 2H, Ar–H), 7.68 (s, 1H, –CH=), 4.70 (s, 2H, CH2). 13C
NMR (400 MHz, d, ppm, DMSO-d6) 166.81(C=O, TZD),
164.34 (C=O, –CH2C=O), 160.78 (C=O, TZD), 141.02,
133.13, 128.36, 127.69, 124.43, 115.29 (ArC), 42.13(CH2).
Elemental Anal. Calcd for (C12H7Br2NO5S) C, 32.98; H,
1.61; N, 3.20. Found: C, 32.93; H, 1.60; N, 3.22.
(Z)-2-(5-(furan-2-ylmethylene)-2,4-dioxothizolidin-3-yl)ace-
tic acid (3k) This compound was prepared according to
general procedure and it was obtained as light yellow solid,
mp195–196 °C. IR (KBr, cm-1) 3250 (OH of COOH),
1
1705 (C=O), 1690 (C=O). H NMR (400 MHz, d, ppm,
DMSO-d6) 8.14 (d, 1H, furan), 7.70 (s, 1H, –CH=), 7.25 (d,
1H, furan), 7.75–7.76 (dd, 1H, furan), 4.63 (s, 2H, CH2).
13C NMR (400 MHz, d, ppm, DMSO-d6) 169.30 (C=O,
TZD), 167.27 (C=O, –CH2C=O), 166.12 (C=O, TZD),
149.47, 148.82, 120.83, 119.36, 118.68, 114.06, 45.43
(CH2). Elemental Anal. Calcd for (C10H7NO5S) C, 47.43;
H, 2.79; N, 5.53. Found: C, 47.40; H, 2.77; N, 5.50.
123

Med Chem Res (2014) 23:987–994
993
(Z)-2-(2,4-dioxo-5-(thiophen-2-ylmethylene)thizolidin-3-yl)-
acetic acid (3l) This compound was prepared according
to general procedure and it was obtained as light orange
solid, mp190–192 °C. IR (KBr, cm-1) 3340 (OH of
COOH), 1725 (C=O), 1695 (C=O). 1H NMR (400 MHz, d,
ppm, DMSO-d6) 8.17 (s, 1H, –CH=), 8.14 (d, 1H, thio-
phene), 7.80 (d, 1H, thiophene), 7.33 (dd, 1H, thiophene),
4.68 (s, 2H, CH2) 13C NMR (400 MHz, d, ppm, DMSO-d6)
169.43 (C=O, TZD), 167.19 (C=O, –CH2C=O), 166.05
(C=O, TZD), 137.22, 136.23, 135.13, 129.43, 126.82,
119.11, 45.37 (CH2). Elemental Anal. Calcd for
(C10H7NO4S2) C, 44.60; H, 2.62; N, 5.20. Found: C, 44.61;
H, 2.60; N, 5.19.
antifungal assay. A set of tubes containing only inoculated
broth was used as controls. For the antibacterial assay after
incubation for 24 h at 37 1 °C and after incubation for
48 h at 25 1 °C for antifungal assay, the tube with no
growth of microorganism was recorded to represent the
MIC expressed in lg/ml. Every experiment in the anti-
bacterial and antifungal assays was replicated twice.
In vitro assay for cytotoxic activity
In vitro cytotoxicity was determined using a standard MTT
assay (Mosmann, 1983) with protocol appropriate for the
individual test system. Test compounds were prepared
prior to the experiment by dissolving in 0.1 % DMSO and
diluted with medium. The cells were then exposed to dif-
ferent concentrations of the drugs. Cells in the control
wells received the same volume of medium containing
0.1 % DMSO. After 24 h, the medium was removed and
cell cultures were incubated with 100 lM MTT reagent
(1 mg/ml) for 5 h at 37 °C. The suspension was placed on
microvibrator for 10 min and absorbance was recorded by
the ELISA reader. The experiment was performed in
triplicate.
(Z)-2-(5-((1-methyl-1H-pyrrol-2-yl)methylene-2,4-dioxo-
thizolidin-3-yl)acetic acid (3m) This compound was
prepared according to general procedure and it was
obtained as orange solid, mp185–186 °C. IR (KBr, cm-1
)
1
3370 (OH of COOH), 1720 (C=O), 1680 (C=O). H NMR
(400 MHz, d, ppm, DMSO-d6) 7.65 (s, 1H, –CH=), 7.29 (d,
1H, pyrrol), 6.61 (d, 1H, pyrrol), 6.36 (dd, 1H, pyrrol), 4.59
(s, 2H, CH2), 3.80 (s, 3H, CH3); 13C NMR (400 MHz, d,
ppm, DMSO-d6): 168.63 (C=O, TZD), 167.34 (C=O,
–CH2C=O), 166.25 (C=O, TZD), 130.88, 127.86, 121.09,
116.71, 113.97, 111.36, 45.81(CH2), 38.87 (N–CH3). Ele-
mental Anal. Calcd for (C11H10N2O4S) C, 49.62; H, 3.79;
N, 10.52. Found: C, 49.60; H, 3.78; N, 10.50.
Acknowledgments Authors are grateful to Dr. A. D. Taranalli,
Principal, for providing necessary facilities. We wish to express our
sincere gratitude to Dr. F. V. Manvi, Dean Faculty of pharmacy and
Dr. C. K. Kokate, Vice-Chancellor for their motivation. Authors are
also grateful to the NMR Research center, IISC, Bangalore, India, for
providing the spectral data.
Microbiology
For the antibacterial and antifungal activity, the com-
pounds were dissolved in dimethylsulfoxide (DMSO).
Further dilutions of the compounds and standard drugs in
the test medium were prepared at the required quantities of
128, 64, 32, 16, 8, 4, 2, and 1 lg/ml concentrations with
Mueller–Hinton broth and Sabouraud dextrose broth. The
MICs were determined using the twofold serial dilution
technique (National Committee for Clinical Laboratory
Standards, 2000). A control test was also performed con-
taining inoculated broth supplemented with only DMSO at
the same dilutions used in our experiments and found
inactive in the culture medium.
References
Albuquerque JFC, Albuquerque A, Azevedo CC, Thomasson F,
Galdino SL, Cantegrel J, Catanho MTJ, Pitta IR, Luu-Duc C
(1995) Substituted thiazolidinediones and thio-imidazolidinon-
es:synthesis, structural study and pharmacological activity.
Pharmazie 50:387–389.
Aydemir N, Bilaloglu R (2003) Genotoxicity of two anticancer drugs,
gemcitabine and topotecan, in mouse bone marrow in vivo.
Mutat Res 537:43–51
Ayhan-Kilcigil G, Altanlar N (2000) Synthesis of 3-substituted
phenacyl-5[2-phenyl-4H-4-oxo-1-benzopyran-6-ylmethyl]-thia-
zolidine-2,4-diones and evaluation of their antimicrobial activ-
ity. Arzneimittelforschung 50:154–157
In vitro assay for antibacterial and antifungal activity
Boyd AS (2007) Thiazolidinedione in dermatology. Int J Derm
46:557–563
Bozdag-Dundar O, Ozgen O, Mentese A, Altanlar N, Atli O, Kendi E,
Ertan R (2007) Synthesis and antimicrobial activity of some new
thiazolyl thiazolidine-2,4-dione derivatives. Bioorg Med Chem
15:6012–6017
Bruno G, Costantino L, Curinga C, Maccari R, Monforte F, Nicolo F,
Ottana R, Vigorita MG (2002) Synthesis and aldose reductase
inhibitory activity of 5-arylidine-2,4-thiazolidinediones. Bioorg
Med Chem 10:1077–1084
The cultures were obtained from Mueller–Hinton broth for
all the bacterial strains after 24 h of incubation at
37 1 °C. Fungi were maintained in Sabouraud dextrose
broth after incubation for 24 h at 25 1 °C. Testing was
carried out in Mueller–Hinton broth and Sabouraud dex-
trose broth at pH 7.4 and the twofold serial dilution tech-
nique was applied. The final inoculum size was 105 CFU/
ml for the antibacterial assay and 104 CFU/ml for the
Elte JWF, Blickle JF (2007) Thiazolidinediones for the treatment of
type 2 diabetes. Eur J Intern Med 18:18–25
123

994
Med Chem Res (2014) 23:987–994
Ertl P, Rohde B, Selzer P (2000) Fast calculation of molecular polar
surface area as a sum of fragment-based contributions and its
application to the prediction of drug transport properties. J Med
Chem 43:3714–3717. (http://www.molinspiration.com/services)
Heerding DA, Christmann LT, Clark TJ, Holmes DJ, Rittenhouse SF,
Takata DT, Venslavsky JW (2003) New benzylidenethiazolidine-
dione as antibacterial agents. Bioorg Med Chem Lett 13:3771–3773
Heneka MT, Landreth GE (2007) Biochim Biophys Acta 1771:
1031–1045
Jiang C, Ting AT, Seed B (1998) PPAR-c agonists inhibits production
monocyte inflammatory cytokines. Nature 391:82–86
Kalaitzidis RG, Sarafidis PA, Bakeis GL (2009) Effects of thiazolid-
inediones beyond glycaemic control. Curr Pharm Des 15:
529–536
Kurebayashi S, Xu X, Ishii S, Shiraishi M, Kouhara H, Kasayama S
(2005) A novel thiazolidinedione MCC-555 down-regulates
tumor necrosis factor-a-induced expression of vascular cell
adhesion molecule-1 in vascular endothelial cells. Atherosclero-
sis 182:71–77
Maccari R, Ottana R, Curinga C, Vigorita MG, Rakowitz D, Steindl
T, Langer T (2005) Structure-activity relationships and molec-
ular modelling of 5-arylidene-2,4-thiazolidinedione active as
aldose reductase inhibitors. Bioorg Med Chem 13:2809–2823
McIntyre RS, Soczynska JK, Woldeyohannes HO, Lewis GF, Leiter
LA, MacQueen GM, Miranda A, Fulgosi D, Konarski JZ,
Kennedy SH (2007) Thiazolidinedione: novel treatments for
cognitive deficits in mood disorders. Expert Opin Pharmacother
8:1615–1628
National Committee for Clinical Laboratory Standards (2000) Method
for dilution antimicrobial susceptibility tests for bacteria that
grow aerobically. Approved standard document M-7: A5.
National Committee for Clinical Laboratory Standards
Pallas 3.7.1.2 (2010) ADME-Tox software, CompuDrug International
Inc. U.S.A.
Prashanth Kumar BR, Karvekar MD, Adhikary L, Nanjan NJ, Suresh
B (2006) Microwave induced synthesis of the thiazolidine-2,4-
dione motif and the efficient solvent free-solid phase parallel
syntheses of 5-benzylidene-thiazolidine-2,4-dione and 5-benzy-
lidene-2-thioxo-thiazolidine-4-one compounds.
Chem 43:897–903
J Heterocycl
Rakowitz D, Maccari R, Ottana R, Vigorita MG (2006) In vitro aldose
reductase inhibitory activity of 5-benzyl-2,4-thiazolidinediones.
Bioorg Med Chem 14:567–574
Ricote M, Li AC, Willson TM, Kelly CJ, Glass CK (1998) The
peroxisome proliferator-activated receptor c is a negative
regulator of macrophage activation. Nature 391:79–82
Tan SF, Ang KP, Fong YF (1986) (Z)-and (E)-arylmethylenehydantoins:
spectroscopic properties and configuration assignment. J Chem Soc
Perkin Trans-2 1941–1944. doi:10.1039/P29860001941
Tuncbilek M, Altanlar N (2006) Synthesis of new 3-(substituted
phenacyl)-5-[3-(4H-4-oxo-1-benzopyran-2-yl)-benzylidene]-2,4-
thiazolidinediones and their antimicrobial activity. Arch Pharm
Chem Life Sci 339:213–216
Vogeli U, Philipsborn W, Nagarajan K, Nair MD (1978) Structures of
addition products of acetylenedicarboxylic acid esters with
various dinucleophiles. An application of C, H-spin-coupling
constants. Helv Chim Acta 61:607–617
Mosmann T (1983) Rapid colorimetric assay for cellular growth and
survrvial. Application to proliferation and cytotoxicity assays.
J Immunol Methods 65:55–63
123