New thiazolidine-2,4-diones as antimicrobial and cytotoxic agent

Article abstract of DOI:10.1007/s00044-011-9876-x

New (Z)-5-substituted-2,4-thiazolidinediones (3a-m) were easily prepared by the condensation of thiazolidine- 2,4-dione (1) with suitable aldehydes (2a-m) via microwave irradiation technique. The reaction between (Z)-5-substituted-2,4- thiazolidinediones and 4-(bromomethyl) benzoic acid, using potassium carbonate as base in refluxing acetone, followed by a workup in acidic medium provided 4-(((Z)-5-substituted-2,4-dioxothiazolidin-3- yl)methyl) benzoic acid derivatives (4a-m). The structures of the newly synthesized compounds were confirmed by IR, ¹H NMR, ¹³C NMR spectral studies, and elemental analysis. All compounds were evaluated for their in vitro antimicrobial and cytotoxic activities. Antibacterial and antifungal results revealed that most of the compounds showed significant activity where as compounds 4c and 4g are found to be broad spectrum antibacterial and antifungal properties, the MIC values were observed in the range of 2-4 and 2-8 μg/ ml, respectively. InMTTcytotoxicity studies, the compound 4g was found most potent. In HeLa, HT29, A549, and MCF-7 cells, the IC₅₀ values were observed in the range of 30-36 μM. Springer Science+Business Media, LLC 2011.

Full text of DOI:10.1007/s00044-011-9876-x

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res
DOI 10.1007/s00044-011-9876-x
ORIGINAL RESEARCH
New thiazolidine-2,4-diones as antimicrobial and cytotoxic agent
•
Shankar G. Alegaon Kallanagouda R. Alagawadi
Received: 6 May 2011 / Accepted: 4 November 2011
Ó Springer Science+Business Media, LLC 2011
Abstract New (Z)-5-substituted-2,4-thiazolidinediones
(3a–m) were easily prepared by the condensation of thia-
zolidine-2,4-dione (1) with suitable aldehydes (2a–m) via
microwave irradiation technique. The reaction between
(Z)-5-substituted-2,4-thiazolidinediones and 4-(bromo-
methyl) benzoic acid, using potassium carbonate as base in
refluxing acetone, followed by a workup in acidic medium
provided 4-(((Z)-5-substituted-2,4-dioxothiazolidin-3-
yl)methyl) benzoic acid derivatives (4a–m). The structures
of the newly synthesized compounds were confirmed by IR,
1H NMR, 13C NMR spectral studies, and elemental analysis.
All compounds were evaluated for their in vitro antimicro-
bial and cytotoxic activities. Antibacterial and antifungal
results revealed that most of the compounds showed signif-
icant activity where as compounds 4c and 4g are found to be
broad spectrum antibacterial and antifungal properties, the
MIC values were observed in the range of 2–4 and 2–8 lg/
ml, respectively. In MTT cytotoxicity studies, the compound
4g was found most potent. In HeLa, HT29, A549, and MCF-7
cells, the IC50 values were observed in the range of
30–36 lM.
Introduction
The development of new antimicrobial and anticancer
therapeutic agents is one of the fundamental goals in
medicinal chemistry. Cytotoxicity and genotoxicity of
anticancer drugs to the normal cells are major problems in
cancer therapy and engender the risk of inducing secondary
malignancy (Aydemir and Bilaloglu, 2003). A dose of
anticancer drug sufficient to kill tumor cells is often toxic
to the normal tissue and leads to many side effects, which
in turn, limits its treatment efficacy. In recent years, there
has been a concerned search for the discovery and devel-
opment of novel selective antimicrobial and anti-tumor
agents, devoid of many of the unpleasant side effects of
conventional antimicrobial and antitumor agents. Peroxi-
some proliferator-activated receptors (PPARs) are mem-
bers of the nuclear hormone receptor superfamily which
are ligand-activated transcription factors. So for, three
PPAR isotopes have been reported: PPARa, PPARb, and
PPARc. Originally, PPAR activity was thought to be lim-
ited to lipid metabolism and glucose homeostasis. Later
studies showed that PPAR activation regulates inflamma-
tory responses, cell proliferation and differentiation, as
well as apoptosis (Houseknecht et al., 2002; Sung et al.,
2004).
Keywords Thiazolidinedione Á Antibacterial activity Á
Antifungal activity Á Cytotoxic activity Á Drug likeliness
Thiazolidine-2,4-dione (TZDs) are a class of insulin
sensitizing drugs which include ciglitazone, pioglitazone,
and rosiglitazone. Apart from their known antidiabetic
activity, the ability of TZDs to contribute to cancer therapy
has been evidenced by numerous in vitro and in vivo studies
(Takashima et al., 2001; Galli et al., 2004; Yoshizumi et al.,
2004; Betz et al., 2005; Turturro et al., 2004; Shiau et al.,
2005; Han and Roman 2006; Kaminskyy et al., 2009;
Li et al., 2010; Patil et al., 2010), antimicrobial and anti-
cancer (Mohsen et al., 1985), antibacterial and antifungal
Electronic supplementary material The online version of this
article (doi:10.1007/s00044-011-9876-x) contains supplementary
material, which is available to authorized users.
S. G. Alegaon (&) Á K. R. Alagawadi
Department of Pharmaceutical Chemistry, KLE University’s
College of Pharmacy, Belgaum, Karnataka 590 010, India
e-mail: shankar_alegaon@yahoo.co.in; sgalegaon@gmail.com
123

Med Chem Res
(Ayhan-Kilcigil and Altanlar, 2000; Heerding et al., 2003;
Tuncbilek and Altanlar, 2006; Bozdag-Dundar et al., 2007;
Mentese et al., 2009; Alegaon and Alagawadi, 2011)
activity. While TZDs are known to stimulate PPAR-c
receptor, they also have multiple PPAR-c independent
effects and the specific role of PPAR-c activation in the
anticancer effects of TZDs is still under investigation. It has
been reported that there exists of a 3 order-of-magnitude
discrepancy between the concentration required to mediate
antitumor effects and that for PPAR-c activation (Wei et al.,
2009). Thus, the dose required for anticancer activity of
thiazolidine-2,4-diones would be significantly lower
than that required to bring hypoglycemic activity. We
report herein the synthesis, characterization and investiga-
tion of antimicrobial and cytotoxic properties of 4-((-5-
benzylidene-2,4-dioxothiazolidin-3-yl)methyl)benzoic acid
derivatives.
confirmed by IR, 1H NMR, 13C NMR spectral studies, and
elemental analysis. The IR spectra of 4-((-5-benzylidene-
2,4-dioxothiazolidin-3-yl)methyl)benzoic acid derivatives
4a–m exhibited a very broad absorption band in the region
of 3,380–2,570 cm-1 attributable to the stretching vibra-
1
tion of carboxylic OH. The H NMR spectra revealed that
two singlets at the region of d 4.61–5.02 ppm and d
7.67–8.12 ppm were assigned to the methylene protons of
N-CH2 and 5-methylidene protons in all title compounds,
respectively. The 13C NMR spectral analyses were con-
sistent with the assigned structures. No large differences
were found in 13C chemical shift for methylene carbon of
N-CH2 (d 43.31–47.38 ppm) and 5-methylidene carbon
(d 130.20–137.77 ppm) in all title compounds. In addition,
in 13C NMR spectra, besides two signals due to the reso-
nances of 2-and 4-carbonylic groups of the thiazolidine-
2,4-dione ring at d 164.5–169.8 ppm, another singlet
attributable to the resonance of the carboxylic carbon was
present in the same range.
Results and discussion
Pharmacology
Chemistry
Antibacterial and antifungal activity
The synthesis of (Z)-4-(5-arylidene-2,4-dioxothizolidin-3-
yl) methyl benzoic acids 4a–m was achieved through the
versatile and efficient synthetic route outlined in Scheme 1.
The starting material thiazolidine-2,4-dione 1 was prepared
according to earlier reported method (Prashanth Kumar
et al., 2006) with minor modification. Further, the con-
densation of compound 1 with the appropriate substituted
aldehydes, using piperidine as base in refluxing toluene for
15 h, provided compounds 3a–m. The reaction time was
significantly reduced (10 min) by using microwave (MW)
irradiation (200 psi, 700 W maximum powers) at 140°C.
The introduction of 5-aryl or 5-heteroarylidene moieties
provided only Z isomers, as already demonstrated by X-ray
diffraction studies (Bruno et al., 2002; Ottana et al., 2005).
The reaction between (Z)-5-arylidene-2,4-thiazolidin-
ediones 3a–m and 4-(bromomethyl) benzoic acid, using
potassium carbonate as base, provided (Z)-4-(5-arylidene-
2,4-dioxothizolidin-3-yl) methyl benzoic acids 4a–m.
The structures of the newly synthesized compounds were
The newly synthesized compounds were evaluated for their
in vitro antibacterial activity against Escherichia coli
(ATCC 35218), Pseudomonas aeruginosa (ATCC 25619),
Staphylococcus aureus (ATCC 25923), Enterococcus fae-
calis (ATCC 35550), antifungal activity against Candida
albicans (ATCC 2091), Aspergillus flavus (NCIM No.
524), Aspergillus niger (ATCC 6275), and Cryptococcus
neoformans (Clinical isolate) by using the twofold serial
dilution technique (National Committee for Clinical Lab-
oratory Standards, 2006) and results are summarized in
Table 1. Ampicillin and ciprofloxacin were used as the
reference standard for antibacterial activity while keto-
conazole was used as the reference standard for antifungal
activity.
In the light, interesting antimicrobial activities of (Z)-4-
(5-arylidene-2,4-dioxothizolidin-3-yl) methyl benzoic acid
derivatives inhibited the growth of bacteria with MIC
values ranging between 2 and 128 lg/ml and showed
COOH
O
O
O
N
NH
NH
H
(a)
(b)
Ar CHO
2 a-m
S
+
S
S
O
O
O
Ar
H
Ar
1
4a-m
3a-m
Scheme 1 Preparation of 4-(((Z)-5-substituted-2,4-dioxothiazolidin-3-yl)methyl)benzoic acid derivatives 4a–m: reagents: a piperidine, acetic
acid, toluene, 110°C for 15 h or MW irradiation 700 W, 10 min, 75–83%; b BrCH2C6H4COOH, K2CO3, acetone, reflux, 48–72 h, 50–62%
123

Med Chem Res
Table 1 Results of antibacterial and antifungal activities of compounds 4a–m [minimum inhibitory concentration (MIC) values in lg/ml]
COOH
O
N
S
O
Ar
H
4a-m
Compound
Ar
E. c
P. a
S. a
E. f
C. a
C. n
A. f
A. n
4a
4
8
128
64
16
16
32
16
4b
4
4
128
128
16
16
16
32
Br
4c
4
4
2
4
2
4
4
4
F
4d
16
16
32
32
64
64
32
64
H3C
4e
8
8
16
16
32
16
32
32
OCH3
4f
32
32
16
16
16
16
32
32
H3CO
4g
H3CO
H3CO
8
8
8
8
2
4
4
4
OCH3
4h
16
16
64
64
32
16
16
32
N
4i
4j
32
16
32
16
128
32
128
16
32
16
32
16
16
32
16
32
N
N
123

Med Chem Res
Table 1 continued
Compound
Ar
E. c
P. a
S. a
E. f
C. a
C. n
A. f
A. n
4k
16
16
32
64
64
64
32
32
O
S
4l
8
16
16
64
32
64
32
32
8
8
4m
16
16
32
32
16
16
N
Amp.
Cip.
NT
2
NT
2
2
2
NT
NT
2
NT
NT
2
NT
NT
1
NT
NT
2
NT
NT
NT
NT
Ket.
NT
NT
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. g Aspergillus niger; NT not tested; Amp ampicillin; Cip ciprofloxacin; Ket ketoconazole
antifungal activity with MICs between 2 and 64 lg/ml.
According to the antimicrobial studies, most of the com-
pounds showed such activity, albeit lower than their anti-
fungal efficacy. This difference may be due to the differ-
ences between the cell structure of bacteria and yeast.
While the cell wall of fungi contains chitin, the cell wall of
bacteria contains murein (Eweis et al., 2006). In addition,
fungi contain ergosterol in their cell membranes instead of
the cholesterol found in the cell membranes of animals
(Kitamura et al., 1999). According to antibacterial studies,
the efficacy against Gram-negative is higher than Gram-
positive bacteria. Compounds 4a, 4b, 4c, 4e, 4g, and 4l
showed high activity against E. coli and 4a, 4b, 4c, 4e, and
4g showed good activity against P. aeruginosa as com-
pared to other compounds. Out of 13, 11 compounds
showed moderate activity against Gram-positive bacteria.
In addition, 4c and 4g exhibited broad spectrum activity
against all bacterial strains, with MIC values 2–8 lg/ml.
Antifungal results indicated that compounds 4c and 4g
have showed good activity against C. albicans, C. neo-
formans, A. flavus, and A. niger. Compound 4l showed
significant activity against A. flavus and A. niger while
other compounds exhibit moderate activity against all
tested fungal strains.
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. Among the thirteen 4-(((Z)-5-substituted-
2,4-dioxothiazolidin-3-yl)methyl)benzoic acid derivatives,
compound 4g, which was 3,4,5-trimethoxyphenyl, exhib-
ited the good inhibitory activity against HeLa, HT29,
A549, and MCF-7 cell lines, with the inhibitory concen-
tration (IC50) values of 30, 32, 36, and 30 lM, respectively.
Compounds 4a, 4d, 4h, 4i, 4j, 4k, 4l, and 4m showed weak
cytotoxic activities against all tested human tumor cell
lines with IC50 values of 60–100 lM. As shown in Table 2,
compounds 4b, 4c, 4e, and 4f showed moderate cytotoxic
activities against all tested human tumor cell lines with
IC50 values of 30–48 lM. In general, structures with
electron-donating substituents showed marginal activity,
the position of substituents appears to play an important
role in activity too.
Molinspiration calculations and ‘‘Rule of 5’’ properties
The structures of thiazolidine-2,4-diones differ in the sub-
stitution pattern. Therefore, they may be considered analo-
gous allowing consideration of structure–activity
relationships. As can be observed in Table 3, we calculated
clogP and topological polar surface area (TPSA) for all
newly synthesized compounds. cLogP is the partition coef-
ficient between water and octanol as a factor of the lipo-
philicity of molecules (Jensen et al., 2005; Veber et al.,
2002). PSA is defined as the surface sum over all polar atoms;
in particular, TPSA is based on the summation of tabulated
surface contributions of polar fragments (Ertl et al., 2000).
These two properties are commonly used functions for the
determination of cell permeability in transport across
membranes (Irwin and Shoichet, 2005). We used software of
Cytotoxic activity
The MTT [3-(4,5-dimethylthiazo-2-yl)-2,5-diphenyl-tetra-
zolium bromide] cell proliferation assay (Mosmann, 1983)
was used to evaluate cytotoxic activity of the synthesized
compounds against four human cancer cell lines including
HeLa (cervical carcinoma), HT29 (colorectal cancer),
A549 (lung cancer), MCF-7 (breast adenocarcinoma)
cell lines (National Centre for Cell Science NCCS,
Pune, India). The inhibition of the cell proliferation was
123

Med Chem Res
Table 2 Results of cytotoxic activities of compounds 4a–m against
four different human cancer cell lines (IC50, lM)
Table 2 continued
Compound Ar
HeLa HT29 A549 MCF-7
COOH
O
4l
76
88
80
76
N
S
S
O
4m
90
88
80
70
Ar
H
N
4a-m
Cell lines include cervical carcinoma (HeLa), colorectal cancer (HT29),
lung cancer (A549), and breast adenocarcinoma (MCF-7)
Compound Ar
HeLa HT29 A549 MCF-7
Table 3 Molinspiration calculations of the 4-(((Z)-5-substituted-2,4-
dioxothiazolidin-3-yl)methyl)benzoic acid derivatives 4a–m
4a
72
88
30
32
72
90
42
36
78
100
COOH
O
4b
38
34
68
48
N
S
O
Br
4c
38
Ar
H
F
4a-m
4d
100
Compound
LogP
MW
MV
TPSA
H3C
4a
4b
4c
4d
4e
4f
3.05
3.86
3.22
3.50
3.08
3.11
2.68
1.88
1.81
1.71
2.31
2.95
2.27
339
418
357
353
369
369
429
340
340
340
329
345
342
284
302
289
301
310
310
361
280
280
280
266
275
286
87.22
76.37
76.37
76.37
85.60
85.60
104.07
89.26
89.26
89.26
89.51
76.37
81.30
4e
48
48
55
58
OCH3
4f
40
30
62
44
32
78
44
36
80
48
30
90
4g
4h
4i
H3CO
4j
H3CO
H3CO
4g
4h
4k
4l
4m
OCH3
Molinspiration for calculations of clogP, TPSA, and
molecular volume (Ertl et al., 2000). In Table 3, clogP of the
compounds 4a–g are higher than the compounds like 4h–
k and 4m. Whereas, there is no large difference in TPSA.
Therefore, the lipophilicity of 4a–g is higher than those of
4h–k and 4m. The low lipophilicity is known as one of the
common cause of poor cell permeability.
N
4i
4j
88
60
90
80
75
72
100
84
N
O
N
To predict the drug likeliness of the synthesized com-
pounds on the guidelines of Lipinski rule of 5 (Molecular
weight B500, logP B5, HBD B 5 and HBA B 10) study
was carried out using Pallas software (Pallas, 2010); the
result are given in Table 4. The relevance of the synthesized
4k
80
94
76
90
123

Med Chem Res
Conclusions
Table 4 Drug likeliness of the 4-(((Z)-5-substituted-2,4-dioxothiaz-
olidin-3-yl)methyl)benzoic acid derivatives 4a–m
We report the synthesis of 4-((2,4-dioxothiazolidin-3-
yl)methyl)benzoic acid derivatives. All compounds were
evaluated for their in vitro antimicrobial and cytotoxic
activities. Antibacterial and antifungal results revealed that
most of the compounds showed significant activity whereas
compounds 4c and 4g are found to be broad spectrum
antibacterial and antifungal properties, the MIC values were
observed in the range of 2–4 and 2–8 lg/ml, respectively. In
MTT cytotoxicity studies, the compound 4g was found most
potent. In HeLa, HT29, A549, and MCF-7 cells, the IC50
values were observed in the range of 30–36 lM. The results
indicates that some of functional groups such as 4-bromo-
phenyl, 4-fluorophenyl, and 3,4,5-trimethoxyphenyl group
present in these compounds displayed role of biological
activity even most of the compounds have similar physico-
chemical parameters. It can be concluded that a combina-
tion of 4-((2,4-dioxothiazolidin-3-yl) methyl) benzoic acid
and substituted phenyl has caused an enhanced antimicro-
bial and cytotoxic effect and hence they are ideally suited
for further modification to obtain more efficacious antimi-
crobial and cytotoxic compounds.
COOH
O
N
S
O
Ar
H
4a-m
Compound
MW
LogP
HBD
HBA
4a
4b
4c
4d
4e
4f
339
418
357
353
369
369
429
340
340
340
329
345
342
3.06
3.96
3.19
3.45
3.05
2.92
2.77
2.05
1.88
1.88
2.54
2.90
2.32
1
1
1
1
1
1
1
1
1
1
1
1
1
5
5
5
5
6
6
8
6
6
6
6
5
6
4g
4h
4i
4j
4k
4l
Experimental protocols
4m
General conditions
molecules with respect to Lipinski rule of five is as follows.
Molecular weight of the compound is important in drug
action, if the molecular weight increases beyond a limit, the
bulkiness of the compounds also increases, which will
affect the drug action (affect the drug receptor/DNA inter-
actions). Molecular weight of compounds lies between 329
and 429 show that these compounds follows Lipinski rule of
5. So the bulkiness of the compounds is in optimum limit for
the action. Pharmacokinetic property optimization is a
rather complex undertaking that is likely to require changes
in those molecular determinants that are responsible for
binding affinity and specificity like hydrogen bonds.
Hydrogen bond acceptor (HBA) and hydrogen bond donor
(HBD) groups in the compound optimize the drug receptor
interaction. Number of HBA (B10) and HBD (B5) in the
proposed compounds obeys the Lipinski rule of 5, so it may
have some of the compounds good absorption or perme-
ability properties through the biological membrane. Disso-
lution is highly interdependent influences of aqueous
solubility, ionizability (pKa) and lipophilicity (logP). Fur-
thermore, logP is a crucial factor governing passive mem-
brane partitioning, influencing permeability opposite to its
effect on solubility. The logP values of the synthesized
compounds lies in between 2 and 4.
All the chemicals used in this study were purchased from
E. Merck, Fluka and Aldrich. Melting point was determined
by electrothermal melting point apparatus and is uncorrected.
TLC controls were carried out on precoated silica gel plates
(F 254 Merck). The IR spectra were recorded on Nicolet
Impact410FTIR spectrophotometer usingKBr pellets.1H and
13C NMR spectra were recorded on AMX-400, Bruker-400
liquid-stateNMRspectrometerusingtetramethylsilane(TMS)
astheinternalstandardandDMSO-d6 asthesolvent. Chemical
shifts were recorded as d (ppm). Elemental analysis was car-
ried out using a Perkin Elmer 2400-CHN Analyzer. MW
experiments were carried out in a domestic oven (200 psi,
700 W maximum powers). Spectra facilities were carried out
by Sophisticated Analytical Instruments Facility (SAIF)
division of Indian Institute of Science, Bangalore, India.
Chemistry
General method for the synthesis of 4-(5-arylidene-2,4-
dioxothiazolidin-3-yl) methylbenzoic acid (4a–m)
A mixture of (Z)-5-substituted-2,4-thiazolidinediones
(3a–m) (10 mmol) and potassium carbonate (20 mmol) in
123

Med Chem Res
4-[((Z)-5-(4-Methylbenzylidene)-2,4-dioxothiazolidin-
3yl)methyl]benzoic acid (4d)
acetone (100 ml) was refluxed for 45 min. Then 4-(bro-
momethyl) benzoic acid (20 mmol) was slowly added and
the mixture was refluxed for 48–72 h. After evaporation of
the solvent under reduced pressure, the crude solid residue
was dissolved in methanol; the solution was acidified (pH
3) with HCl and was stirred at room temperature for
30 min. After evaporation to dryness in vacuum, the crude
solid was washed with H2O and recrystallized from
methanol/CHCl3 providing pure acid 4a–m.
Yield 58%, mp 262–264°C, IR (KBr, cm-1): 3262 (OH),
1
1723, 1703 (C=O); H NMR (400 MHz, d, ppm, DMSO-
d6): 2.36 (s, 3H, CH3), 4.61 (s, 2H, CH2), 7.25(d, 2H, Ar–
H, J = 7.6 Hz), 7.36 (d, 2H, Ar–H, J = 8.1 Hz), 7.48 (d,
2H, Ar–H, J = 6.8 Hz), 7.56 (d, 2H, Ar–H, J = 6.5 Hz),
7.95 (s, 1H, –CH=), 12.57 (s, 1H, OH); 13C NMR
(100 MHz, d, ppm, DMSO-d6): 21.07, 43.31, 114.00,
119.66, 130.01, 130.24, 133.75, 137.77, 141.20, 157.38,
164.33, 165.19, 167.06; Elemental Anal. Calcd for
C19H15NO4S (353): C, 64.58; H, 4.28; N, 3.96. Found: C,
64.55; H, 4.25; N, 3.97.
4-[((Z)-5-Benzylidene-2,4-dioxothiazolidin-
3yl)methyl]benzoic acid (4a)
Yield 62%, mp 275–277°C, IR (KBr, cm-1): 3276 (OH),
1
1720, 1695 (C=O); H NMR (400 MHz, d, ppm, DMSO-
4-[((Z)-5-(3-Methoxybenzylidene)-2,4-dioxothiazolidin-
3yl)methyl]benzoic acid (4e)
d6): 4.90 (s, 2H, CH2), 7.42 (d, 2H, Ar–H, J = 8.2 Hz),
7.49–7.56 (m, 3H, Ar–H), 7.62 (d, 2H, Ar–H, J = 7.4 Hz),
7.92 (d, 2H, Ar–H, J = 8 Hz), 7.97 (s, 1H, –CH=), 12.94
(s, 1H, OH); 13C NMR (100 MHz, d, ppm, DMSO-d6):
44.37, 121.07, 127.59, 129.34, 129.64, 130.12, 130.21,
130.70, 132.86, 133.51, 140.16, 165.43, 166.90, 167.29;
Elemental Anal. Calcd for C18H13NO4S (339): C, 63.71; H,
3.86; N, 4.13. Found: C, 63.67; H, 3.85; N, 4.11.
Yield 55%, mp 256–258°C, IR (KBr, cm-1): 3330 (OH),
1
1720, 1700 (C=O); H NMR (400 MHz, d, ppm, DMSO-
d6): 3.97 (s, 3H, OCH3), 4.89 (s, 2H, CH2), 7.05–7.19 (m,
2H, Ar–H), 7.40–7.47 (m, 2H, Ar–H), 7.55 (d, 2H, Ar–H,
J = 8.1 Hz), 7.90 (s, 1H, –CH=),7.93 (d, 2H, Ar–H,
J = 8.3 Hz), 12.99 (s, 1H, OH); 13C NMR (100 MHz, d,
ppm, DMSO-d6): 44.36, 55.27, 115.44, 116.60, 121.95,
127.56, 129.63, 130.46, 133.48, 134.22, 140.16, 159.62,
165.38, 166.89, 167.24; Elemental Anal. Calcd for
C19H15NO5S (369): C, 61.78; H, 4.09; N, 3.79. Found: C,
61.73; H, 4.04; N, 3.75.
4-[((Z)-5-(4-Bromobenzylidene)-2,4-dioxothiazolidin-
3yl)methyl]benzoic acid (4b)
Yield 52%, mp 262–263°C, IR (KBr, cm-1): 3320 (OH),
1
1720, 1700 (C=O); H NMR (400 MHz, d, ppm, DMSO-
4-[((Z)-5-(4-Methoxybenzylidene)-2,4-dioxothiazolidin-
3yl)methyl]benzoic acid (4f)
d6): 5.02 (s, 2H, CH2), 7.47 (d, 2H, Ar–H, J = 8.1 Hz),
7.55 (d, 2H, Ar–H, J = 8.3 Hz), 7.65 (d, 2H, Ar–H,
J = 8.2 Hz), 7.81 (d, 2H, Ar–H, J = 8.3 Hz), 8.03(s, 1H,
–CH=), 13.09 (s, 1H, OH); 13C NMR (100 MHz, d, ppm,
DMSO-d6): 46.44, 116.34, 125.37, 126.53, 127.32, 128.02,
129.66, 130.98, 132.34, 149.67, 154.55, 165.76, 167.46,
168.33; Elemental Anal. Calcd for C18H12BrNO4S (418):
C, 51.69; H, 2.89; N, 3.35. Found: C, 51.65; H, 2.86; N,
3.33.
Yield 58%, mp 265–267°C, IR (KBr, cm-1): 3325 (OH),
1
1725, 1700 (C=O); H NMR (400 MHz, d, ppm, DMSO-
d6): 3.84 (s, 3H, OCH3), 4.91 (s, 2H, CH2), 7.11 (d, 2H,
Ar–H, J = 7.3 Hz), 7.41(d, 2H, Ar–H, J = 8.1 Hz), 7.60
(d, 2H, Ar–H, J = 6.7 Hz), 7.92 (d, 2H, Ar–H,
J = 6.5 Hz), 7.94 (s, 1H, –CH=), 12.95 (s, 1H, OH); 13C
NMR (100 MHz, d, ppm, DMSO-d6): 44.26, 55.48, 114.96,
117.69, 125.32, 127.52, 129.62, 132.02, 132.30, 133.57,
140.28, 151.24, 165.53, 166.88, 167.34; Elemental Anal.
Calcd for C19H15NO5S (369): C, 61.78; H, 4.09; N, 3.79.
Found: C, 61.75; H, 4.05; N, 3.74.
4-[((Z)-5-(4-Fluorobenzylidene)-2,4-dioxothiazolidin-
3yl)methyl]benzoic acid (4c)
Yield 60%, mp 250–252°C, IR (KBr, cm-1): 3255 (OH),
1
1725, 1700 (C=O); H NMR (400 MHz, d, ppm, DMSO-
4-[((Z)-5-(3,4,5-Trimethoxybenzylidene)-2,4-
dioxothiazolidin-3yl)methyl]benzoic acid (4g)
d6): 4.90 (s, 2H, CH2), 7.36–7.42 (m, 4H, Ar–H), 7.70 (d,
2H, Ar–H, J = 8.3 Hz), 7.90 (d, 2H, Ar–H, J = 6.5 Hz),
7.98 (s, 1H, –CH=), 12.94 (s, 1H, OH); 13C NMR
(100 MHz, d, ppm, DMSO-d6): 47.38, 115.58, 124.36,
125.55, 127.32, 127.98, 128.79, 130.53, 131.88, 148.37,
155.16, 165.37, 166.27, 168.65; Elemental Anal. Calcd for
C18H12FNO4S (357): C, 60.50; H, 3.38; N, 3.92. Found: C,
60.47; H, 3.36; N, 3.90.
Yield 50%, mp 260–262°C, IR (KBr, cm-1): 3360 (OH),
1722, 1704 (C=O); 1H NMR (400 MHz, d, ppm, DMSO-d6):
3.72 (s, 3H, OCH3), 3.82 (s, 6H, OCH3), 4.90 (s, 2H, CH2),
6.95 (s, 2H, Ar–H), 7.42 (d, 2H, Ar–H, J = 8.3 Hz), 7.91 (s,
1H, –CH=), 7.92 (d, 2H, Ar–H, J = 6.7 Hz), 12.94 (s, 1H,
123

Med Chem Res
OH); 13C NMR (100 MHz, d, ppm, DMSO-d6): 44.60,
54.56, 54.98, 115.34, 118.65, 126.48, 127.34, 128.21,
131.62, 132.16, 133.20, 141.32, 152.45, 165.02, 166.43,
168.18; Elemental Anal. Calcd for C21H19NO7S (429): C,
58.73; H, 4.46; N, 3.26. Found: C, 58.70; H, 4.44; N, 3.23.
d6): 4.87 (s, 2H, CH2), 6.75 (q, 1H, J = 1.7, J = 1.6 Hz,
furan), 7.15 (d, 1H, J = 3.4 Hz, furan), 7.40 (d, 2H, Ar–H,
J = 8.2 Hz), 7.78 (s, 1H, –CH=), 7.91 (d, 2H, Ar–H,
J = 8.2 Hz), 8.07 (d, 1H, J = 1.4 Hz, furan), 12.93 (s, 1H,
OH); 13C NMR (100 MHz, d, ppm, DMSO-d6): 44.50,
120.21, 123.33, 125.42, 127.33, 128.13, 129.69, 131.55,
140.72, 157.64, 162.44, 165.28, 166.23, 168.65; Elemental
Anal. Calcd for C16H11NO5S (329): C, 58.35; H, 3.37; N,
4.25. Found: C, 58.32; H, 3.33; N, 4.26.
4-[((Z)-2,4-Dioxo-5-((pyridine-2-yl)methylene)thiazolidin-
3yl)methyl]benzoic acid (4h)
Yield 58%, mp 275–276°C, IR (KBr, cm-1): 3355 (OH),
1
1725, 1700 (C=O); H NMR (400 MHz, d, ppm, DMSO-
4-[((Z)-2,4-Dioxo-5-((thiophen-2-yl)methylene)thiazolidin-
3yl)methyl]benzoic acid (4l)
d6): 4.90 (s, 2H, CH2), 7.42 (d, 2H, Ar–H, J = 8.1 Hz),
7.53–7.67 (m, 4H, pyridine), 7.89 (d, 2H, Ar–H,
J = 6.5 Hz), 7.97 (s, 1H, –CH=), 12.95 (s, 1H, OH); 13C
NMR (100 MHz, d, ppm, DMSO-d6): 45.32, 119.25,
120.24, 124.63, 125.34, 128.46, 129.08, 130.20, 132.12,
140.30, 151.86, 164.36, 166.02, 167.45; Elemental Anal.
Calcd for C17H12N2O4S (340): C, 59.99; H, 3.55; N, 8.23.
Found: C, 59.96; H, 3.52; N, 8.25.
Yield 62%, mp 258–260°C, IR (KBr, cm-1): 3320 (OH),
1
1720, 1695 (C=O); H NMR (400 MHz, d, ppm, DMSO-
d6): 4.90 (s, 2H, CH2), 7.41–7.43 (d, 1H, J = 3.7 Hz, thi-
ophene), 7.60 (d, 1H, J = 1.6 Hz, thiophene), 7.76 (d, 2H,
J = 8.2 Hz, Ar–H), 7.79–7.91 (m, 2H, Ar–H and 1H, thi-
ophene), 7.92 (s, 1H, –CH=), 12.75 (s, 1H, OH); 13C NMR
(100 MHz, d, ppm, DMSO-d6): 44.55, 119.65, 123.60,
124.63, 127.63, 128.84, 129.61, 130.29, 139.98, 158.83,
161.38, 165.11, 166.87, 167.25; Elemental Anal. Calcd for
C16H11NO4S2 (345): C, 55.64; H, 3.21; N, 4.06. Found: C,
55.63; H, 3.20; N, 4.02.
4-[((Z)-2,4-Dioxo-5-((pyridine-3-yl)methylene)thiazolidin-
3yl)methyl]benzoic acid (4i)
Yield 56%, mp 258–260°C, IR (KBr, cm-1): 3370 (OH),
1723, 1700 (C=O); 1H NMR (400 MHz, d, ppm, DMSO-d6):
4.89 (s, 2H, CH2), 7.12 (d, 2H, Ar–H, J = 8.1 Hz), 7.21–7.24
(m, 2H, pyridine), 7.39 (d, 2H, pyridine, J = 5.1 Hz), 7.90 (d,
2H, Ar–H, J = 6.5 Hz), 7.92 (s, 1H, –CH=), 12.93 (s, 1H,
OH); 13C NMR (100 MHz, d, ppm, DMSO-d6): 45.54,
118.86, 121.56, 123.54, 125.87, 127.48, 129.77, 130.43,
133.54, 135.86, 145.22, 151.33, 165.40, 166.28, 168.06;
Elemental Anal. Calcd for C17H12N2O4S (340): C, 59.99; H,
3.55; N, 8.23. Found: C, 59.95; H, 3.53; N, 8.25.
4-[((Z)-5-((1-Methyl-1H-pyrrol-2-yl)methylene)-2,4-
dioxothiazolidin-3yl)methyl]benzoic acid (4m)
Yield 55%, mp 255–256°C, IR (KBr, cm-1): 3340 (OH),
1
1725, 1699 (C=O); H NMR (400 MHz, d, ppm, DMSO-
d6): 3.80 (s, 3H, CH3), 4.99 (s, 2H, CH2), 6.35 (d, 1H,
J = 3.1 Hz, pyrrol), 7.28 (m, 2H, Ar–H and 1H, pyrrol),
7.60 (m, 2H, Ar–H and 1H, pyrrol), 7.65 (s, 1H, –CH=),
14.30 (s, 1H, OH); 13C NMR (100 MHz, d, ppm, DMSO-
d6): 32.76, 44.82, 120.92, 122.76, 125.82, 127.44, 128.02,
129.55, 131.56, 137.53, 157.44, 162.43, 165.43, 167.35,
168.63; Elemental Anal. Calcd for C17H14N2O4S (342): C,
59.64; H, 4.12; N, 8.18. Found: C, 59.62; H, 4.10; N, 8.19.
4-[((Z)-2,4-Dioxo-5-((pyridine-4-yl)methylene)thiazolidin-
3yl)methyl]benzoic acid (4j)
Yield 50%, mp 270–272°C, IR (KBr, cm-1): 3325 (OH),
1
1726, 1704 (C=O); H NMR (400 MHz, d, ppm, DMSO-
d6): 4.71 (s, 2H, CH2), 7.20–7.28 (m, 4H, pyridine), 7.50
(d, 2H, Ar–H, J = 8.1 Hz), 7.92 (d, 2H, Ar–H,
J = 6.8 Hz), 8.12 (s, 1H, –CH=), 12.42 (s, 1H, OH); 13C
NMR (100 MHz, d, ppm, DMSO-d6): 44.85, 118.43,
120.98, 122.87, 125.64, 127.55, 129.70, 131.11, 135.80,
149.45 150.01, 165.40, 166.24 167.87; Elemental Anal.
Calcd for C17H12N2O4S (340): C, 59.99; H, 3.55; N, 8.23.
Found: C, 59.95; H, 3.53; N, 8.25.
Antimicrobial activity
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 inoculums size was 105 CFU/
ml for the antibacterial assay and 104 CFU/ml for the
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
4-[((Z)-5-((Furan-2-yl)methylene)-2,4-dioxothiazolidin-
3yl)methyl]benzoic acid (4k)
Yield 62%, mp 263–265°C, IR (KBr, cm-1): 3280 (OH),
1
1720, 1700 (C=O); H NMR (400 MHz, d, ppm, DMSO-
123

Med Chem Res
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The experiment was performed in triplicate. The IC50
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