Antimycobacterial evaluation of novel hybrid arylidene thiazolidine-2,4- diones

Article abstract of DOI:10.1016/j.bmcl.2014.01.007

A series of novel hybrid heterocycles comprising arylidene thiazolidine-2,4-dione and 1-cyclopropyl-2-(2-fluorophenyl)ethanone were synthesized. These compounds were evaluated for their antimycobacterial activity against Mycobacterium tuberculosis H₃₇Rv in High Throughput Screen. Most of the hybrid arylidene thiazolidine-2,4-diones displayed moderate to good activity with MIC of less than 50 μM. Compound 1m exhibited maximum potency being 5.87 fold more active at EC₅₀ and 6.26 fold more active at EC₉₀ than the standard drug pyrimethamine.

Full text of DOI:10.1016/j.bmcl.2014.01.007

Bioorganic & Medicinal Chemistry Letters 24 (2014) 1089–1093
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Antimycobacterial evaluation of novel hybrid arylidene
thiazolidine-2,4-diones
c,d,e
Singanan Ponnuchamy a, Selvaraj Kanchithalaivan b, Raju Ranjith Kumar b, , Mohamed Ashraf Ali
,
⇑
Tan Soo Choon c
a Orchid Chemicals & Pharmaceuticals Ltd, R&D Center, Chennai 600119, India
b Department of Organic Chemistry, School of Chemistry, Madurai Kamaraj University, Madurai 625021, Tamil Nadu, India
c Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Minden 11800, Penang, Malaysia
d New Drug Discovery Research, Department of Medicinal Chemistry, Alwar Pharmacy College, Alwar, Rajasthan 301030, India
e New Drug Discovery Research, Department of Medicinal Chemistry, Sunrise University, Alwar, Rajasthan 301030, India
a r t i c l e i n f o
a b s t r a c t
Article history:
A series of novel hybrid heterocycles comprising arylidene thiazolidine-2,4-dione and 1-cyclopropyl-2-
(2-fluorophenyl)ethanone were synthesized. These compounds were evaluated for their antimycobacte-
rial activity against Mycobacterium tuberculosis H37Rv in High Throughput Screen. Most of the hybrid
arylidene thiazolidine-2,4-diones displayed moderate to good activity with MIC of less than 50 lM.
Compound 1m exhibited maximum potency being 5.87 fold more active at EC50 and 6.26 fold more active
Received 2 October 2013
Revised 31 December 2013
Accepted 6 January 2014
Available online 11 January 2014
at EC90 than the standard drug pyrimethamine.
Keywords:
Ó 2014 Elsevier Ltd. All rights reserved.
Thiazolidine-2,4-dione
Knoevenagel condensation
Antimycobacterial activity
Mycobacterium tuberculosis
High Throughput Screen
Tuberculosis (TB) is a chronic bacterial infection caused by
Mycobacterium Tuberculosis bacteria (MTB), which spreads through
air and usually infects the lungs. TB can be treated by taking sev-
eral drugs for a period of 6–9 months. Of the approved drugs, the
first-line anti-TB agents that form the core of treatment regimen
include isoniazid (INH), rifampin (RIF), ethambutol (EMB) and pyr-
azinamide (PZA). Regimens for treating TB disease have an initial
phase of 2 months, followed by a choice of several options for
the continuation phase of either 4 or 7 months. The initial empiric
treatment of TB starts with a 4-drug regimen: isoniazid, rifampin,
pyrazinamide and either ethambutol or streptomycin. Once the TB
isolate is known to be fully susceptible, ethambutol (or streptomy-
cin, if it is used as a fourth drug) can be discontinued.1 After
2 months of therapy (for a fully susceptible isolate), pyrazinamide
can be stopped. Isoniazid and rifampin are continued as daily or
intermittent therapy for 4 more months. If isolated isoniazid resis-
tance is documented, isoniazid is discontinued and treatment with
rifampin, pyrazinamide and ethambutol is continued for the entire
6 months. The Directly Observed Therapy (DOT) is the most recom-
mended for all patients. With DOT, patients on the above regimens
can be switched to 2–3 times per week dosing after an initial
2 weeks of daily dosing.2
One of the most significant methods for the development of
highly active compounds is the combination of active pharmaco-
phores into a single unit.3 The biological screening of such
compounds may result in new lead compounds with better activity
than the standard drugs. In this context, we envisage to investigate
the antimycobacterial activity of novel hybrid thiazolidine-2,4-
diones 1 (Fig. 1) that comprise 5-arylidene-thiazolidine-2,4-dione
and 1-cyclopropyl-2-(2-fluorophenyl)ethanone unit. The latter is
the key component of the platelet inhibitor prasugrel 2.4 Inciden-
tally, thiazolidine-2,4-dione derivatives have been identified as
the privileged class of organic heterocycles with profound biologi-
cal activities. The well known drugs used for the treatment of type
II diabetes mellitus troglitazone 35 and pioglitazone 46 comprise
thiazolidine-2,4-dione moiety (Fig. 1). Troglitazone 37 has been
shown to exhibit anticancer activities through the inhibition of
the Raf/MEK/ERK signaling pathway. Further, the 5-arylidene thia-
zolidine-2,4-dione derivatives have been identified as potent and
highly selective inhibitors of the PIM kinase.8 However, the
antimycobacterial activity of these heterocyclic derivatives are still
unknown. The above importance of 5-arylidene thiazolidine-2,4-
diones and in view of our continued interest in the synthesis and
biological evaluation of novel heterocycles,9 we herein report the
⇑
Corresponding author. Tel.: +91 9655591445.
E-mail address: raju.ranjithkumar@gmail.com (R. Ranjith Kumar).
0960-894X/$ - see front matter Ó 2014 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.bmcl.2014.01.007

1090
S. Ponnuchamy et al. / Bioorg. Med. Chem. Lett. 24 (2014) 1089–1093
O
F
O
N
S
R
O
1
O
F
O
HN
S
S
O
N
O
OAc
O
S
OH
2
3
, Troglitazone
, Prasugrel
O
O
OEt
OH
HN
HN
S
O
, Pioglitazone
N
O
O
4
5, Inhibitor of PIM-2 kinase
Figure 1. Design of the target molecule.
preliminary results of the antimycobacterial activity of novel 3-
[2-cyclopropyl-1-(2-fluorophenyl)-2-oxoethyl]-5-[(Z)-(aryl)meth-
ylidene]-1,3-thiazolane-2,4-diones 1.
In the present investigation, the starting material 2-bromo-1-
cyclopropyl-2-(2-fluorophenyl)ethanone 7 was synthesized from
the reaction of 2-bromo-2-(2-fluorophenyl)acetonitrile and cyclo-
yields. Further, the presence of sterically hindered groups in the
aldehyde and bulky indoline, quinoxaline and benzo[d][1,3]dioxaz-
ole too had no adverse effects in the yield of the product. However,
the reaction failed to occur with aliphatic aldehydes. The structure
of all the arylidene thiazolidine-2,4-dione hybrid heterocycles 1a–
q were elucidated with the help of elemental analysis, mass, IR and
NMR spectroscopic techniques. Moreover, the NMR spectra of 1
and thiazolidine-2,4-dione 8 were similar except for the presence
of new signals in the 1H and 13C NMR spectra of 1 due to the aro-
matic ring and benzylidene protons and carbons. Further, the
DEPT-135 spectrum of 1 reveals the absence of one ‘CH2’ carbon
and appearance of new signals due to aromatic ‘C’ and ‘CH’
carbons. The structure of 1 assigned from NMR spectroscopy was
further solved from single crystal X-ray studies. The ORTEP dia-
gram of 1f reveals (Z)-configuration for the exocyclic alkene
(Fig. 2).13
propyl magnesium bromide solution, following
a literature
report.10 Further, the reaction of 7 with thiazolidine-2,4-dione 6
in DMF in the presence of NaHCO3 at ambient temperature affor-
ded novel 3-(2-cyclopropyl-1-(2-fluorophenyl)-2-oxoethyl)thia-
zolidine-2,4-dione 8 in 93% yield (Scheme 1).11 The structure of 8
is in complete agreement with its elemental analysis, mass, IR
and NMR spectroscopy. The mass spectrum of 8 has a characteristic
molecular ion peak at 292.0 (M+). The IR spectrum of 8 shows
strong absorptions at 1765, 1703 and 1687 cmÀ1 which are due
to the three carbonyl groups. Further, in the 1H NMR spectrum of
8 the diastereotopic 5-CH2 protons appeared as doublets at 3.94
and 4.01 ppm with J value 17.7 Hz. The N–CH appeared as a singlet
at 6.40 ppm whereas the cyclopropyl ring protons were observed
as multiplets between 0.96 and 1.92 ppm. In the 13C NMR of 8,
the carbonyls of the thiazolidine ring appeared at 170.4 and
170.5 ppm whilst the signal at 200.8 ppm is due to the carbonyl
of the 1-cyclopropyl-2-(2-fluorophenyl)ethanone moiety.
All the hybrid thiazolidine-2,4-diones 1a–q and
screened for their in vitro antimycobacterial activity against
Mycobacterium tuberculosis 37RV (MTB-H37Rv) in High
8 were
H
a
Throughput Screen (HTS) using an assay adapted from the micro-
dilution alamar Blue (AB) broth assay reported by Collins and
Franzblau14 and additionally an alternative method for end-point
detection using the Promega reagent BacTiter-Glo™ Microbial
Cell Viability (BTG). Five standard drugs were used as references
together with the synthesized compound for the assay. Data were
analyzed using the IDBS Activity Base software and the dose
response result was analyzed using a four parameter logistic fit
to the data (Excel Fit equation 205) with the maximum and
minimum locked at 100 and 0. From these curves, EC50 and
EC90 values were calculated. The MIC is the minimum concentra-
tion of the compound required to inhibit 90% of bacterial growth
and MIC’s of the compounds are given in Table 2 along with
standard drugs for comparison.
In the next step, the synthesis of novel (Z)-5-benzylidene-
3-(2-cyclopropyl-1-(2-fluorophenyl)-2-oxoethyl)thiazolidine-2,4-
diones 1 via the Knoevenagel condensation of 8 with various
aromatic aldehydes was performed (Scheme 1).12 The reaction
proceeded in isopropyl alcohol in the presence of piperidine as
base under reflux condition affording 1a–q in excellent yields
(73–88%). A total of seventeen novel arylidene thiazolidine-2,4-
dione hybrid heterocycles were synthesized (Table 1). The reaction
occurred smoothly with aromatic aldehydes bearing electron-
withdrawing or electron-donating group affording 1 in excellent
Ar
1
5
S
1
2
Br
F
S
S
5
O
O
4
2
N3
ArCHO
DMF
NaHCO3, rt
6-7 h
O
O
O
O
4
N
H
N3
F
iPrOH
O
piperidine
reflux, 4-5h
6
O
F
7
O
8
1
Scheme 1. Synthesis of 1,3-thiazolane-2,4-diones 1.

S. Ponnuchamy et al. / Bioorg. Med. Chem. Lett. 24 (2014) 1089–1093
1091
Table 1
Yield and melting point of hybrid arylidene thiazolidine-2,4-diones 1 and 8
Entry
Compd
Aldehyde
Product
Yield (%)
82
Mp (°C)
O
O
CHO
1
1a
134–135
F
O
N
S
S
O
Cl
CHO
F
O
2
3
4
5
6
7
8
9
1b
1c
1d
1e
1f
79
82
79
84
84
74
87
87
181–183
154–156
165–168
127–128
142–143
199–202
75–79
N
O
Cl
F
CHO
O
O
O
O
O
O
O
F
O
N
S
S
S
S
S
S
S
O
F
HO
MeO
MeS
CHO
F
O
N
O
OH
OMe
SMe
CHO
F
O
N
O
CHO
F
O
N
O
HO
HO
CHO
F
O
1g
1h
1i
N
OH
OH
O
MeO
MeO
MeO
CHO
F
O
N
OMe
OMe
O
OMe
CHO
OMe
F
O
OMe
142–143
N
MeO
O
OMe
MeO
MeO
MeO
O
CHO
F
O
N
10
1j
88
121–123
OMe
OMe
S
O
MeO
(continued on next page)

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S. Ponnuchamy et al. / Bioorg. Med. Chem. Lett. 24 (2014) 1089–1093
Table 1 (continued)
Entry
Compd
Aldehyde
HO
Product
Yield (%)
Mp (°C)
O
O
MeO
CHO
CHO
F
O
Br
N
11
12
1k
82
76–79
Br
S
S
O
OMe
HO
F
O
N
N
1l
80
159–163
O
N
S
CHO
O
O
F
O
13
14
1m
1n
81
73
94–98
N
S
S
S
O
S
CHO
F
O
Br
N
120–121
S
O
Br
CHO
O
F
N
15
1o
81
161–164
N
O
H
N
S
O
N
H
CHO
O
O
F
O
16
17
18
1p
1q
8
78
86
93
194–196
156–158
94–95
N
N
S
S
O
OHC
O
O
F
O
N
O
O
O
O
F
O
NA
N
S
O
All the hybrid thiazolidine-2,4-diones 1a–q and 8 showed EC50
<25 and were more potent than the standard drug pyrimeth-
amine (EC50 = 37.35). Seven compounds 1c–e, 1h, 1m, 1q and 8
antimycobacterial activity when compared to the other
substituents.
From the data in Table 2 it is evident that twelve compounds
with EC50 value 11, 10, 11, 8, 6, 9 and 6
found to be more potent than the standard drug cycloserine
(EC50 = 12.47 M). Two compounds 1m and were nearly
equipotent of which 1m comprising a thiophene ring was the
most active with EC50 of 6 M and was 1.95 and 5.85 times more
l
M, respectively, were
1c–h, 1l, 1m, 1o–q and 8 with MIC of 25 lM were found to be
equipotent to cycloserine and 4 times more potent than pyrimeth-
amine. However, none of these compounds were found to be active
than other standard drugs. The hybrid thiazolidine-2,4-diones
1a–q and 8 were also tested for cytotoxicity in VERO cells at
l
8
l
potent than the standard drugs cycloserine and pyrimethamine,
concentrations of 62.5 lg/mL. After 72 h of exposure, viability
respectively. Further, three compounds 1d, 1h and 1m with
EC90 value 19, 16 and 11 lM, respectively, were found to be more
active than pyrimethamine. These observations clearly showed
that the presence of unsubstituted thiophene ring in the hybrid
thiazolidine-2,4-diones makes remarkable improvement in
was assessed on the basis of cellular conversion of MTT into a for-
mazan product using the Promega CellTiter 96 non-radioactive cell
proliferation assay. All the compounds were found to be non-toxic
up to 62.5
lg/mL. The compounds were bactericidal and the
cytotoxic profile was within the acceptable range.

S. Ponnuchamy et al. / Bioorg. Med. Chem. Lett. 24 (2014) 1089–1093
1093
Instrument Facility, Indian Institute of Technology Madras for
single crystal X-ray diffraction studies.
Supplementary data
Supplementary data associated with this article can be found,
in the online version, at http://dx.doi.org/10.1016/j.bmcl.2014.
01.007.
References and notes
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Suhadev, M.; Sakthivel, R.; Narayanan, P. R. Am. J. Respir. Crit. Care Med. 2010,
181, 743.
3. (a) Liu, A.; Huang, L.; Wang, Z.; Luo, Z.; Mao, F.; Shan, W.; Xie, J.; Lai, K.; Li, X.
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2011, 54, 6704; (c) Solomon, V. R.; Hu, C.; Lee, H. Bioorg. Med. Chem. 2009, 17,
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Gottlieb, S.; Neumann, F.-J.; Ardissino, D.; De Servi, S.; Murphy, A. A.;
Riesmeyer, J.; Weerakkody, G.; Gibson, C. M.; Antman, E. M. N. Eng. J. Med.
2007, 357, 2001; (b) Baker, W. L.; White, C. M. Am. J. Cardiovasc. Drugs 2009, 9,
213.
5. Minamikawa, J.; Tanaka, S.; Yamauchi, M.; Inoue, D.; Koshiyama, H. J. Clin.
Endocrinol. Metab. 1998, 83, 1818.
6. Momose, Y.; Mequro, K.; Ikeda, H.; Hatanaka, C.; Oi, S.; Sohda, T. Chem. Pharm.
Bull. 1991, 39, 1440.
7. (a) Yao, C.-J.; Lai, G.-M.; Chan, C.-F.; Cheng, A.-L.; Yang, Y.-Y.; Chuang, S.-E. Int. J.
Cancer 2006, 118, 773; (b) Yan, K.-H.; Yao, C.-J.; Chang, H.-Y.; Lai, G.-M.; Cheng,
A.-L.; Chuang, S.-E. Mol. Carcinog. 2010, 49, 235.
8. Dakin, L. A.; Block, M. H.; Chen, H.; Code, E.; Dowling, J. E.; Feng, X.; Ferguson, A.
D.; Green, I.; Hird, A. W.; Howard, T.; Keeton, E. K.; Lamb, M. L.; Lyne, P. D.;
Pollard, H.; Read, J.; Wua, A. J.; Zhang, T.; Zheng, X. Bioorg. Med. Chem. Lett.
2012, 22, 4599.
9. (a) Jeyachandran, V.; Ranjith Kumar, R.; Ali, M. A.; Choon, T. S. Bioorg. Med.
Chem. Lett. 2013, 23, 2101; (b) Sivakumar, S.; Ranjith Kumar, R.; Ali, M. A.;
Choon, T. S. Eur. J. Med. Chem. 2013, 65, 240.
10. Pan, X.; Huang, R.; Zhang, J.; Ding, L.; Li, W.; Zhang, Q.; Liu, F. Tetrahedron Lett.
2012, 53, 5364.
11. Procedure for the synthesis of 8: A mixture of 1,3-thiazolidine-2,4-dione (6,
1 mmol), 2-bromo-1-cyclopropyl-2-(2-fluorophenyl)ethanone (7, 1 mmol) and
sodium bicarbonate (2 equiv) were dissolved in DMF and stirred at ambient
temperature (27–28 °C) for 7 h. After completion of the reaction as evidenced
by TLC (eluent 7:3/hexane–ethyl acetate) the reaction mixture was poured into
water (25 mL), extracted with ethyl aceate and the solvent distilled under
Figure 2. ORTEP diagram of 1f.
Table 2
HTS Antimycobacterial activity of 1 and 8
Compd
EC50
(lM)
EC90
(lM)
MIC (lM)
1a
1b
1c
1d
1e
1f
1g
1h
1i
1j
1k
1l
1m
1n
1o
1p
17
25
11
10
11
16
21
8
13
19
15
24
6
18
18
14
9
6
0.07
36
55
25
19
27
25
55
16
27
53
54
36
11
27
27
68
21
26
0.08
50
50
25
25
25
25
25
25
50
50
50
25
25
50
25
25
1q
8
25
25
0.16
25.00
12.50
0.31
Amikacin
Cycloserine
Ethambutol
Isoniazid
Pyrimethamine
12.47
<1.56
0.18
13.49
32.79
0.29
37.35
74.96
100.00
vacuum
to
obtain
pure
8.
3-(2-Cyclopropyl-1-(2-fluorophenyl)-2-
oxoethyl)thiazolidine-2,4-dione 8: Isolated as white crystalline powder. Yield:
93%; mp 94–95 °C; 1H NMR (300 MHz, CDCl3) dH: 0.96–1.03 (m, 2H), 1.16–1.30
(m, 2H), 1.85–1.9 (m, 1H), 3.98–4.00 (s, 2H), 6.39 (s, 1H), 7.10–7.61 (m, 4H); 13
C
In conclusion, the present work reports a simple synthesis of
novel hybrid heterocycles comprising biologically active compo-
nents viz. arylidene thiazolidine-2,4-dione and 1-cyclopropyl-2-
(2-fluorophenyl)ethanone. The structural features of these
compounds were established with the help of NMR and single
crystal X-ray studies. In addition, all these hybrid thiazolidine-
2,4-diones were screened for their antimycobacterial activity
against MTB H37Rv in High Throughput Screen wherein compound
1m was found to be the most active.
NMR (75 MHz, CDCl3) dC: 12.4, 13.0, 18.9, 33.3, 59.3, 115.6 (2JC,F = 22 Hz), 119.7
(2JC,F = 13 Hz), 123.9 (4JC,F = 4 Hz), 131.1 (3JC,F = 8 Hz), 132.7, 161.3
(1JC,F = 248 Hz), 170.4, 170.8, 200.8; IR (KBr)
m
cmÀ1 1687, 1703, 1764; MS
(m/z) [MÀ1] 292. Anal. Calcd for C14H12FNO3S: C, 57.33; H, 4.12; N, 4.78.
Found: C, 57.38; H, 4.15; N, 4.77.
12. Synthesis of (Z)-5-benzylidene-3-(2-cyclopropyl-1-(2-fluorophenyl)-2-oxoethyl)-
thiazolidine-2,4-diones 1. General procedure: A mixture of 8 (1 mmol), aromatic
aldehyde (1 mmol) and piperdine (2 equiv.) in isopropyl alcohol was refluxed
for 4–5 h. After completion of the reaction as seen from the TLC, (eluent 7:3/
hexane–ethyl acetate) the mixture was allowed to cool to room temperature.
Then the reaction mixture was poured onto ice-water and neutralized with
dilute HCl. The precipitated solid was filtered, washed with water and dried
under
vacuum.
(Z)-3-(2-Cyclopropyl-1-(2-fluorophenyl)-2-oxoethyl)-5-(4-
Acknowledgments
(methylthio)benzylidene)thiazolidine-2,4-dione (1f): Isolated as yellow powder.
Yield: 84%; mp 142–143 °C; 1H NMR (400 MHz, DMSO-d6) dH: 0.95–0.99 (m,
2H), 1.00–1.03 (m, 2H), 2.17–2.23 (m, 1H), 2.50 (s, 3H), 6.48 (s, 1H), 7.23–7.30
(m, 2H), 7.39–7.42 (m, 2H), 7.44–7.49 (m, 2H), 7.57 (d, J = 6.3 Hz, 2H), 7.95 (s,
1H); 13C NMR (100 MHz, DMSO-d6) dC: 12.3, 12.4, 14.0, 18.6, 59.5, 115.5
(2JC,F = 21 Hz), 118.4, 120.0 (2JC,F = 14.0 Hz), 124.2 (4JC,F = 2 Hz), 125.7, 128.8,
130.7, 131.2 (3JC,F = 9 Hz), 131.7, 134.1, 160.6 (1JC,F = 246 Hz), 164.8, 166.4,
R.R.K., S.P. and S.K. thank the University Grants Commission,
New Delhi for funds through Major Research Project F. No. 42-
242/2013 (S.R.) and Department of Science and Technology, New
Delhi for funds under (i) DST-SERC Fast Track Scheme No. SR/FT/
CS-073/2009 and (ii) IRHPA program for the high resolution NMR
facility in the Department. S.P. thanks the management, Orchid
Chemicals and Pharmaceuticals Ltd, Chennai, for providing labora-
tory facilities. R.R.K. and S.P. thank the Sophisticated Analytical
201.0; IR (KBr)
22H18FNO3S2: C, 61.81; H, 4.24; N, 3.28. Found: C, 61.84; H, 4.24; N, 3.32.
m
cmÀ1 1688, 1710, 1737; MS (m/z) [M+1] 428. Anal. Calcd for
C
13. Suresh, J.; Venkateshan, M.; Ponnuchamy, S.; Ranjith Kumar, R.; Lakshman, P. L.
N. Acta Crystallogr. 2013, E69, 188.
14. Colins, L.; Franzblau, S. G. Antimicrob. Agents Chemother. 1997, 41, 1004.