Synthesis and in vitro antiproliferative and antibacterial activity of new thiazolidine-2,4-dione derivatives

Article abstract of DOI:10.1080/14756366.2017.1387543

In our present research, we synthesised new thiazolidine-2,4-diones (12–28). All the newly synthesised compounds were evaluated for antiproliferative and antibacterial activity. Antiproliferative evaluation was carried out using normal human skin fibroblasts and tumour cell lines: A549, HepG2, and MCF-7. The IC₅₀ values were determined for tested compounds revealing antiproliferative activity. Moreover, safety index (SI) was calculated. Among all tested derivatives, the compound 18 revealed the highest antiproliferative activity against human lung, breast, and liver cancer cells. More importantly, the derivative 18 showed meaningfully lower IC₅₀ values when compared to the reference substance, irinotecan, and relatively high SI values. Moreover, newly synthesised compounds were screened for the bacteria growth inhibition in vitro. According to our screening results, most active compound was the derivative 18 against Gram-positive bacteria. Therefore, it may be implied that the novel compound 18 appears to be a very promising agent for anticancer treatment.

Full text of DOI:10.1080/14756366.2017.1387543

Journal of Enzyme Inhibition and Medicinal Chemistry
ISSN: 1475-6366 (Print) 1475-6374 (Online) Journal homepage: http://www.tandfonline.com/loi/ienz20
Synthesis and in vitro antiproliferative and
antibacterial activity of new thiazolidine-2,4-dione
derivatives
Nazar Trotsko, Agata Przekora, Justyna Zalewska, Grażyna Ginalska, Agata
Paneth & Monika Wujec
To cite this article: Nazar Trotsko, Agata Przekora, Justyna Zalewska, Grażyna Ginalska, Agata
Paneth & Monika Wujec (2018) Synthesis and in vitro antiproliferative and antibacterial activity of
new thiazolidine-2,4-dione derivatives, Journal of Enzyme Inhibition and Medicinal Chemistry, 33:1,
17-24, DOI: 10.1080/14756366.2017.1387543
To link to this article: http://dx.doi.org/10.1080/14756366.2017.1387543
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Date: 06 November 2017, At: 01:24

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY, 2017
VOL. 33, NO. 1, 17–24
https://doi.org/10.1080/14756366.2017.1387543
RESEARCH PAPER
Synthesis and in vitro antiproliferative and antibacterial activity of new
thiazolidine-2,4-dione derivatives
a
b
b
b
a
a
_
Nazar Trotsko , Agata Przekora , Justyna Zalewska , Grazyna Ginalska , Agata Paneth and Monika Wujec
b
aDepartment of Organic Chemistry, Faculty of Pharmacy, Medical University of Lublin, Lublin, Poland; Department of Biochemistry and
Biotechnology, Faculty of Pharmacy, Medical University of Lublin, Lublin, Poland
ABSTRACT
ARTICLE HISTORY
Received 26 August 2017
Revised 18 September 2017
Accepted 24 September 2017
In our present research, we synthesised new thiazolidine-2,4-diones (12–28). All the newly synthesised
compounds were evaluated for antiproliferative and antibacterial activity. Antiproliferative evaluation was
carried out using normal human skin fibroblasts and tumour cell lines: A549, HepG2, and MCF-7. The IC50
values were determined for tested compounds revealing antiproliferative activity. Moreover, safety index
(SI) was calculated. Among all tested derivatives, the compound 18 revealed the highest antiproliferative
activity against human lung, breast, and liver cancer cells. More importantly, the derivative 18 showed
meaningfully lower IC50 values when compared to the reference substance, irinotecan, and relatively high
SI values. Moreover, newly synthesised compounds were screened for the bacteria growth inhibition in
vitro. According to our screening results, most active compound was the derivative 18 against Gram-posi-
tive bacteria. Therefore, it may be implied that the novel compound 18 appears to be a very promising
agent for anticancer treatment.
KEYWORDS
Antiproliferative activity;
IC50; thiazolidinediones;
thiosemicarbazones; anti-
bacterial activity
Introduction
found3–6. Researchers’ interest in the derivatives of thiazolidine-
2,4-dione has increased recently, the main reason being a wide
spectrum of biological properties shown by these derivatives. It
has been confirmed by numerous reviews on the activity and
Cancer is one of the most serious health problems in the world. It
is the second cause of death after heart diseases worldwide.
According to the World Cancer Report 2014, 8.2 million people
died from cancer in 20121.
Lung, liver, stomach, and bowel cancer are the most common
causes of human deaths worldwide, accounting for nearly a half of
all cancer deaths. The five most common types of disease diag-
nosed in 2012 were lung, prostate, colorectal, stomach, and liver
cancer among men; and breast, colorectal, lung, cervix, and stom-
ach cancer among women1.
Despite enormous efforts aimed at the implementation of new
treatment strategies of chemotherapeutic agents, treatment results
in most cases are unsatisfactory2. Therefore, there is an urgent need
to find new classes of substances with selective action against
tumour cells.
mechanisms of action of thiazolidine-2,4-diones2,7,8
.
Thiazolidine-2,4-diones are known as antidiabetic drugs and
include rosiglitazone, pioglitazone, and darglitazone. Moreover,
thiazolidine-2,4-dione derivatives possess biological activities such
as aldose reductase inhibitory9, antibacterial6,10–13, antifungal10,
antitubercular14, and anti-inflammatory activity15, etc.
Regulation of both cell proliferation and pathways of apoptosis
connected with cell death is important in understanding various
diseases including malignancies16. Therefore, identification of regu-
lators of the cell cycle and apoptosis stimulators is an attractive
strategy to explore potential anticancer agents17.
There are several mechanisms of anticancer activity of thiazoli-
dine-2,4-dione derivatives already discussed in the literature. The
Heterocyclic compounds play an important role in cancer ther-
apy. Among them, the derivatives of thiazolidine-2,4-dione are most known ones are induction of apoptosis, cell differentiation,
ꢀ
Department of Organic Chemistry, Faculty of Pharmacy, Medical University of Lublin, Chodzki 4A, 20-093
CONTACT Nazar Trotsko
Lublin, Poland
nazar.trotsko@umlub.pl
Supplemental data for this article can be accessed here.
ß 2017 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distri-
bution, and reproduction in any medium, provided the original work is properly cited.

18
N. TROTSKO ET AL.
and cell cycle arrest8. The thiazolidine-2,4-dione derivatives show- Germany). Elemental analyses were performed by AMZ 851 CHX
ing anticancer activity are mainly derivatives modified in the pos-
ition 5 of the thiazolidine-2,4-dione ring.
analyser and the results were within 0.4% of the theoretical
value.
Patil et al. reported that 10 new derivatives of 5-benzylidene-
thiazolidine-2,4-dione had displayed a variable degree of antiproli-
ferative activity against seven tumour cell lines. 2-{4-[(2,4-dioxo-1,
3-thiazolidin-5-ylidene)methyl]phenoxy}-N-[3-(trifluoromethyl)phenyl]
acetamide showed the most significant effect against breast cancer
MCF-7 and leukaemia K562 cell lines3.
General procedure for the synthesis of hydrazones (12–21) or thio-
semicarbazones (22–28)
Anhydrous ethanol (5–10 ml) was added to the mixture of
0.001 mol hydrazide (3-chlorobenzhydrazide, 2,4-dichlorobenzhy-
New 5-substituted pyrazoline thiazolidine-4-one was tested for drazide), thiosemicarbazide or appropriate 4-substituted thiosemi-
carbazide, and 0.001 mol corresponding compounds5–11. Then, the
anticancer activity according to the NCI protocol. The most active
compounds were the derivatives of 2-(2,4-dioxo-1,3-thiazolidin-5-ylide-
ne)acetic acid, namely 5-{2-[5-(2-hydroxyphenyl)-3-(4-methoxyphenyl)-
4,5-dihydro-1H-pyrazol-1-yl]-2-oxoethylidene}-1,3-thiazolidine-2,4-dione
and 5-{2-[5-(2-hydroxyphenyl)-3-(naphthalen-2-yl)-4,5-dihydro-1H-
mixture was heated under reflux till the dissolving of the sub-
strates. The reflux lasted 5–15 min but in cases of compounds 13,
14, 17, 28 reflux lasted 1.5 h. After that, the mixture was cooled
and precipitate was filtered off, dried, and crystallised from buta-
nol or acetic acid.
pyrazol-1-yl]-2-oxoethylidene}-1,3-thiazolidine-2,4-dione.
These
derivatives displayed the highest inhibitory activity against leukae-
mia cells (HL-60 (TB)) and CNS cancer (SF-295)5.
3-{[2-(3-Chlorobenzoyl)hydrazinylidene]methyl}phenyl (2,4-dioxo-
1,3-thiazolidin-5-yl)acetate (12). Yield 87%, m.p. 244–245 ꢀC, 1H
NMR d ppm (DMSO-d6): 3.45–3.47 m (2H, CH–CH2); 4.86–4.90 m
(1H, CH–CH2); 7.20–7.23 m, 7.51–7.70 m, 7.89 d, 7.97 s (8H, Ar,
J ¼ 7.5 Hz); 8.45 s (1H, CH¼N); 12.03 s (1H, NHCO); 12.16 s (1H, NH,
thiazolidine), 13 C NMR d ppm (DMSO-d6): 36.4 (CH2CH), 46.8
(CH2CH), 119.9, 123.9, 126.0, 127.0, 127.8, 130.7, 131.0, 132.2,
133.8, 135.8, 136.4, 147.6 (12 ꢁ Car.), 151.0 (CH¼N), 162.3, 169.6
(2 ꢁ C¼O), 172.7, 175.9 (2 ꢁ C¼O, thiazolidine). MS (APCI) [M]þ m/
z ¼ 432.0441. Anal. calc. for C19H14ClN3O5S (431.85) (%): C 52.84; H
3.27; N 9.73. Found: C 52.80; H 3.24; N 9.70.
A number of derivatives of 2,4-dioxothiazolidine-5-acetic acid
with the ring of 5-substituted-2-amino-1,3,4-thiadiazole showed
cytotoxic effects in vitro against four human tumour cell lines
(cervical carcinoma – HeLa, colorectal cancer – HT29, lung cancer
– A549, and breast cancer – MCF-7). Among the 14 derivatives,
2-(2,4-dioxo-1,3-thiazolidin-5-yl)-N-[5-(3,4,5-trimethoxyphenyl)-1,3,4-
thiadiazol-2-yl]acetamide showed significant inhibitory activity
against the tested cell lines6.
Havrylyuk et al. described 2,4-dioxothiazolidine-5-acetic deriva-
tives with benzoxazole or benzothiazole substituents that showed
in vitro cytotoxic activity against some tumour cell lines: leukaemia
(HL-60(TB), K-562, MOLT-4), breast cancer (MDA-MB-231/ATCC),
CNS cancer (SF-268), melanoma (LOX IMVI), and prostate cancer,
all tested according to the NCI protocol. Among the tested com-
pounds, three were the most active: 3-methoxy-4-(2,4-thiazolidine-
dione-5-acetoxy)benzylidenehydrazone of benzoxazole-2-thioacetic
3-{[2-(2,4-Dichlorobenzoyl)hydrazinylidene]methyl}phenyl
(2,4-
dioxo-1,3-thiazolidin-5-yl)acetate (13). Yield 95%, m.p. 198–200 ꢀC,
1H NMR d ppm (DMSO-d6): 3.44–3.47 m (2H, CH–CH2); 4.85–4.90 m
(1H, CH–CH2); 7.20–7.24 m, 7.52–7.66 m, 7.79 d (7H, Ar, J ¼ 1.8 Hz);
8.27 s (1H, CH¼N); 12.06 s (1H, NHCO); 12.15 s (1H, NH, thiazoli-
dine), 13 C NMR d ppm (DMSO-d6): 36.4 (CH2CH), 46.9 (CH2CH),
120.1, 123.6, 123.9, 126.0, 128.0, 129.1, 129.8, 130.7, 131.2, 132.2,
136.1, 147.6 (12 ꢁ Car.), 151.0 (CH¼N), 162.2, 169.6 (2 ꢁ C¼O),
172.7, 176.0 (2 ꢁ C¼O, thiazolidine). MS (APCI) [M]þ m/
z ¼ 466.0046. Anal. calc. for C19H13Cl2N3O5S (466.29) (%): C 48.94;
H 2.81; N 9.01. Found: C 48.90; H 2.77; N 8.99.
acid,
3-methoxy-4-(2,4-thiazolidinedione-5-acetoxy)benzylidenehy-
drazone of benzothiazole-2-thioacetic acid, and 4-(2,4-thiazolidine-
dione-5-acetoxy)benzylidenehydrazone of benzothiazole-2-thioacetic
acid4.
The aim of the present research was to synthesise new thiazoli-
dine-2,4-dione derivatives and to evaluate in vitro their potential
as anticancer and antibacterial agents. As thiosemicarbazone18–20
and acylhydrazone derivatives20–24 present anticancer activity,
which is similar to the activity of the above mentioned thiazoli-
dine-2,4-diones, it was assumed that the structure modification of
the thiazolidine-2,4-dione ring in position 5 by thiosemicarbazide
and hydrazide derivatives can extend the biological activity of the
new compounds.
4-{[2-(3-Chlorobenzoyl)hydrazinylidene]methyl}phenyl (2,4-dioxo-
1,3-thiazolidin-5-yl)acetate (14). Yield 83%, m.p. 215–216 ꢀC, 1H
NMR d ppm (DMSO-d6): 3.44–3.47 m (2H, CH–CH2); 4.85–4.89 m
(1H, CH–CH2); 7.25 d, 7.81
d
(4H, 4-O–C6H4, J ¼ 8.7 Hz);
7.55–7.70 m, 7.89 d, 7.97 s (4H, Ar, J ¼ 7.5 Hz); 8.46 s (1H, CH¼N);
11.98 s (1H, NHCO); 12.16 s (1H, NH, thiazolidine), 13 C NMR d ppm
(DMSO-d6): 36.4 (CH2CH), 46.8 (CH2CH), 122.7, 127.0, 127.8, 128.9,
131.0, 132.1, 132.6, 133.8, 135.8, 147.8 (12 ꢁ Car.), 151.9 (CH¼N),
162.2, 169.4 (2 ꢁ C¼O), 172.7, 176.0 (2 ꢁ C¼O, thiazolidine). MS
(APCI) [M]þ m/z ¼ 432.0420. Anal. calc. for C19H14ClN3O5S (431.85)
(%): C 52.84; H 3.27; N 9.73. Found: C 52.79; H 3.23; N 9.72.
Experimental
Chemistry
Materials and methods
Melting points were determined using Fisher-Johns apparatus
(Fisher Scientific, Schwerte, Germany) and were not corrected. The
1H NMR and 13 C NMR spectra were recorded by a Bruker Avance
300 MHz instrument using DMSO-d6 as solvent and TMS as an
internal standard. Chemical shifts were expressed as d (ppm). MS
using atmospheric pressure chemical ionisation (APCI) was
recorded on a Bruker MicroTOF II mass spectrometer. APCI settings
were as follows: vaporiser temperature, 350 ꢀC; drying gas tem-
perature, 180 ꢀC; drying gas flow, 4 l/min; and nebuliser pressure,
4-{[2-(2,4-Dichlorobenzoyl)hydrazinylidene]methyl}phenyl
(2,4-
dioxo-1,3-thiazolidin-5-yl)acetate (15). Yield 61%, m.p. 196–198 ꢀC,
1H NMR d ppm (DMSO-d6): 3.44–3.47 m (2H, CH–CH2); 4.85–4.89 m
(1H, CH–CH2); 7.25 d (2H, 4-O–C6H4, J ¼ 8.7 Hz); 7.52–7.66 m, (2H,
Ar); 7.78–7.82 m (3H, Ar); 8.28 s (1H, CH¼N); 12.00 s (1H, NHCO);
12.16 s (1H, NH, thiazolidine), 13 C NMR d ppm (DMSO-d6): 36.4
(CH2CH), 46.8 (CH2CH), 122.7, 127.7, 128.0, 129.0, 131.2, 132.2,
2 bar. The purity of the compounds was checked by TLC on plates 132.4, 134.5, 135.7, 147.7 (12 ꢁ Car.), 151.9 (CH¼N), 162.1, 169.4
with silica gel Si 60 F254, produced by Merck Co. (Darmstadt, (2 ꢁ C¼O), 172.7, 175.9 (2 ꢁ C¼O, thiazolidine). MS (APCI) [M]þ m/

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
19
z ¼ 465.9945. Anal. calc. for C19H13Cl2N3O5S (466.29) (%): C 48.94; 4-{[2-(3-Chlorobenzoyl)hydrazinylidene]methyl}-2-methoxyphenyl
H 2.81; N 9.01. Found: C 48.92; H 2.75; N 9.00.
(2,4-dioxo-1,3-thiazolidin-5-ylidene)acetate (21). Yield 93%, m.p.
246–248 ꢀC, 1H NMR d ppm (DMSO-d6): 3.86 s (3H, OCH3); 7.09 s
(1H, CH¼); 7.30–7.38 m, 7.52–7.70 m, 7.88–7.90 m, 7.97 s (7H, Ar);
8.47 s (1H, CH¼N); 12.02 s (1H, NHCO); 12.98 s (1H, NH, thiazoli-
dine), 13 C NMR d ppm (DMSO-d6): 56.5 (OCH3), 115.8 (CH ¼ C),
110.5, 121.1, 123.6, 127.0, 127.8, 131.0, 132.1, 133.8, 134.2, 135.8,
140.6, 148.0 (12 ꢁ Car.), 146.2 (CH¼C), 151.4 (CH¼N), 162.2, 163.7
(2 ꢁ C¼O), 166.6, 169.4 (2 ꢁ C¼O, thiazolidine). MS (APCI) [M]þ m/
z ¼ 460.0306. Anal. calc. for C20H14ClN3O6S (459.86) (%): C 52.24; H
3.07; N 9.14. Found: C 52.22; H 3.02; N 9.10.
4-{[2-(3-Chlorobenzoyl)hydrazinylidene]methyl}-2-methoxyphenyl
(2,4-dioxo-1,3-thiazolidin-5-yl)acetate (16). Yield 92%, m.p.
220–222 ꢀC, 1H NMR d ppm (DMSO-d6): 3.42–3.44 m (2H, CH–CH2);
3.85 s (3H, OCH3); 4.84–4.88 m (1H, CH–CH2); 7.21 d, 7.31–7.34 m,
7.48–7.70 m, 7.87–7.90 m, 7.97 s (7H, Ar, J ¼ 8.1 Hz); 8.44 s (1H,
CH¼N); 12.00 s (1H, NHCO); 12.14 s (1H, NH, thiazolidine), 13 C NMR
d ppm (DMSO-d6): 36.1 (CH2CH), 46.9 (CH2CH), 56.4 (OCH3), 110.5,
121.1, 123.6, 127.0, 127.8, 131.0, 132.1, 133.8, 133.9, 135.9, 141.0,
148.1 (12 ꢁ Car.), 151.5 (CH¼N), 162.2, 168.8 (2 ꢁ C¼O), 172.7, 175.8
(2 ꢁ C¼O, thiazolidine). MS (APCI) [M]þ m/z ¼ 462.0488. Anal. calc. 3-{[2-(Phenylcarbamothioyl)hydrazinylidene]methyl}phenyl
(2,4-
dioxo-1,3-thiazolidin-5-yl)acetate (22). Yield 86%, m.p. 209–211 ꢀC,
1H NMR d ppm (DMSO-d6): 3.42–3.45 m (2H, CH–CH2); 4.84–4.88 m
(1H, CH–CH2); 7.17–7.25 m, 7.36–7.56 m, 7.74–7.78 m (9H, Ar); 8.16 s
(1H, CH¼N); 10.17 s, 11.89 s (NHCSNH); 12.16 s (1H, NH, thiazoli-
dine), 13 C NMR d ppm (DMSO-d6): 36.4 (CH2CH), 46.8 (CH2CH),
120.3, 123.8, 126.0, 126.4, 126.7, 128.6, 130.4, 136.3, 139.5, 142.2
(12 ꢁ Car.), 151.0 (CH¼N), 169.6 (C¼O), 172.7, 175.9 (2 ꢁ C¼O, thia-
zolidine), 176.7 (C¼S). MS (APCI), fragment ion (m/z): 355.0727;
336.0097; 277.0273; 179.0266; 157.9907; 136.0225; 94.0657. Anal.
calc. for C19H16N4O4S2 (428.48) (%): C 53.26; H 3.76; N 13.08.
Found: C 53.29; H 3.77; N 13.09.
for C20H16ClN3O6S (461.88) (%): C 52.01; H 3.49; N 9.10. Found: C
52.00; H 3.47; N 9.06.
4-{[2-(2,4-Dichlorobenzoyl)hydrazinylidene]methyl}-2-methoxyphenyl
(2,4-dioxo-1,3-thiazolidin-5-yl)acetate (17). Yield 87%, m.p.
186–188 ꢀC, 1H NMR d ppm (DMSO-d6): 3.41–3.44 m (2H, CH–CH2);
3.84 s (3H, OCH3); 4.84–4.87 m (1H, CH–CH2); 7.20 d, 7.32 dd,
7.48–7.58 m, 7.64 d, 7.78 d (6H, Ar, J1¼8.4 Hz, J2 ¼ 1.8 Hz); 8.25 s
(1H, CH¼N); 12.01 s (1H, NHCO); 12.16 s (1H, NH, thiazolidine), 13 C
NMR d ppm (DMSO-d6): 36.0 (CH2CH), 46.9 (CH2CH), 56.4 (OCH3),
110.7, 121.1, 123.6, 128.0, 129.8, 131.2, 132.2, 133.6, 134.5, 135.7,
141.0, 148.0 (12 ꢁ Car.), 151.5 (CH¼N), 162.2, 168.8 (2 ꢁ C¼O),
172.7, 175.8 (2 ꢁ C¼O, thiazolidine). MS (APCI) [M]þ 3-[{2-[(4-Methylphenyl)carbamothioyl]hydrazinylidene}methyl]phenyl
m/m/z ¼ 496.0087. Anal. calc. for C20H15Cl2N3O6S (496.32) (%): C
(2,4-dioxo-1,3-thiazolidin-5-yl)acetate (23). Yield 80%, m.p.
194–196 ꢀC, 1H NMR
d ppm (DMSO-d6): 2.32 s (3H, CH3);
48.40; H 3.05; N 8.47. Found: C 48.37; H 3.03; N 8.44.
3.42–3.45 m (2H, CH–CH2); 4.83–4.87 m (1H, CH–CH2); 7.18 d,
7.39–7.51 m, 7.73–7.77 m (8H, Ar, J ¼ 8.4 Hz); 8.15 s (1H, CH¼N);
10.10 s, 11.84 s (NHCSNH); 12.16 s (1H, NH, thiazolidine), 13 C NMR d
ppm (DMSO-d6): 21.1 (CH3), 36.4 (CH2CH), 46.8 (CH2CH), 120.2,
123.7, 126.4, 126.6, 129.0, 130.4, 135.1, 136.3, 136.9, 142.0
(12 ꢁ Car.), 151.0 (CH¼N), 169.6 (C¼O), 172.7, 175.9 (2 ꢁ C¼O, thia-
zolidine), 176.8 (C¼S). MS (APCI) [M]þ m/z ¼ 443.0791. Anal. calc.
for C20H18N4O4S2 (442.51) (%): C 54.29; H 4.10; N 12.66. Found: C
54.23; H 3.95; N 12.63.
2-{[2-(3-Chlorobenzoyl)hydrazinylidene]methyl}phenyl (2,4-dioxo-
1,3-thiazolidin-5-ylidene)acetate (18). Yield 88%, m.p. 243–245 ꢀC,
1H NMR
d
ppm (DMSO-d6): 7.16 s (1H, CH¼); 7.34–7.46 m,
7.52–7.68 m, 7.84–7.99 m (8H, Ar); 8.51 s (1H, CH¼N); 11.94 s (1H,
NHCO); 12.95 s (1H, NH, thiazolidine), 13 C NMR d ppm (DMSO-d6):
116.5 (CH ¼ C), 123.5, 126.6, 127.0, 127.5, 127.8, 131.0, 131.8, 132.2,
133.8, 134.3, 135.7, 143.1 (12 ꢁ Car.), 145.7 (CH¼C), 148.9 (CH¼N),
162.1, 164.3 (2 ꢁ C¼O), 166.6, 169.3 (2 ꢁ C¼O, thiazolidine). MS
(APCI) [M]þ m/z ¼ 430.0207. Anal. calc. for C19H12ClN3O5S (429.83)
(%): C 53.09; H 2.81; N 9.78. Found: C 53.06; H 2.82; N 9.79.
4-[(2-Carbamothioylhydrazinylidene)methyl]phenyl (2,4-dioxo-1,3-
1
thiazolidin-5-yl)acetate (24). Yield 66%, m.p. 208–210 ꢀC, H NMR d
ppm (DMSO-d6): 3.42–3.45 m (2H, CH–CH2); 4.84–4.88 m (1H,
CH–CH2); 7.17 d, 7.88 d (4H, Ar, J ¼ 8.7 Hz); 8.05 s (2H, NH2); 8.22 s
(1H, CH¼N); 11.46 s (NHCS); 12.16 s (1H, NH, thiazolidine), 13 C NMR
d ppm (DMSO-d6): 36.4 (CH2CH), 46.8 (CH2CH), 122.4, 129.0, 132.7,
141.6 (6 ꢁ Car.), 151.5 (CH¼N), 169.4 (C¼O), 172.7, 175.9 (2 ꢁ C¼O,
thiazolidine), 178.5 (C¼S). MS (APCI) [M]þ m/z ¼ 353.0346. Anal.
calc. for C13H12N4O4S2 (352.39) (%): C 44.31; H 3.43; N 15.90.
Found: C 44.27; H 3.38; N 15.86.
3-{[2-(3-Chlorobenzoyl)hydrazinylidene]methyl}phenyl (2,4-dioxo-
1,3-thiazolidin-5-ylidene)acetate (19). Yield 91%, m.p. 282–283 ꢀC,
1H NMR
d
ppm (DMSO-d6): 7.08 s (1H, CH¼); 7.31–7.34 m,
7.54–7.70 m, 7.88–7.97 m (8H, Ar); 8.47 s (1H, CH¼N); 12.05 s (1H,
NHCO); 12.98 s (1H, NH, thiazolidine), 13 C NMR d ppm (DMSO-d6):
116.6 (CH ¼ C), 120.0, 123.7, 124.0, 126.0, 127.0, 127.9, 130.7, 131.0,
132.2, 133.8, 135.7, 136.4 (12 ꢁ Car.), 147.5 (CH¼C), 150.8 (CH¼N),
162.3, 164.3 (2 ꢁ C¼O), 166.9, 169.6 (2 ꢁ C¼O, thiazolidine). MS
(APCI) [M]þ m/z ¼ 430.0212. Anal. calc. for C19H12ClN3O5S (429.83)
(%): C 53.09; H 2.81; N 9.78. Found: C 53.05; H 2.77; N 9.74.
4-{[2-(Phenylcarbamothioyl)hydrazinylidene]methyl}phenyl
(2,4-
dioxo-1,3-thiazolidin-5-yl)acetate (25). Yield 84%, m.p. 159–162 ꢀC,
1H NMR d ppm (DMSO-d6): 3.43–3.46 m (2H, CH–CH2); 4.85–4.89 m
(1H, CH–CH2); 7.19–7.24 m, 7.38 t, 7.56 d (7H, Ar, J ¼ 7.5 Hz); 7.99 d
(2H, 4-O–C6H4, J ¼ 8.7 Hz); 8.17 s (1H, CH¼N); 10.15 s, 11.86 s
(NHCSNH); 12.16 s (1H, NH, thiazolidine), 13 C NMR d ppm (DMSO-
d6): 36.5 (CH2CH), 46.8 (CH2CH), 122.4, 125.9, 126.5, 128.5, 129.4,
132.5, 139.5, 142.3 (12 ꢁ Car.), 151.7 (CH¼N), 169.4 (C¼O), 172.7,
175.9 (2 ꢁ C¼O, thiazolidine), 176.5 (C¼S). MS (APCI), fragment ion
(m/z): 338.3411; 284.2929; 257.2465; 149.0230. Anal. calc. for
C19H16N4O4S2 (428.48) (%): C 53.26; H 3.76; N 13.08. Found: C
53.22; H 3.77; N 13.08.
4-{[2-(3-Chlorobenzoyl)hydrazinylidene]methyl}phenyl (2,4-dioxo-
1,3-thiazolidin-5-ylidene)acetate (20). Yield 93%, m.p. 278–279 ꢀC,
1H NMR d ppm (DMSO-d6): 7.08 s (1H, CH¼); 7.37 d, 7.55–7.69 m,
7.82–7.91 m, 7.97 s (8H, Ar, J ¼ 8.4 Hz); 8.48 s (1H, CH¼N); 12.00 s
(1H, NHCO); 12.98 s (1H, NH, thiazolidine), 13 C NMR d ppm (DMSO-
d6): 116.7 (CH ¼ C), 122.6, 127.0, 127.8, 128.9, 131.0, 132.1, 132.9,
133.8, 135.8, 147.7 (12 ꢁ Car.), 145.4 (CH¼C), 151.7 (CH¼N), 162.2,
164.1 (2 ꢁ C¼O), 166.5, 169.4 (2 ꢁ C¼O, thiazolidine). MS (APCI)
[M]þ m/z ¼ 430.0210. Anal. calc. for C19H12ClN3O5S (429.83) (%): C
53.09; H 2.81; N 9.78. Found: C 53.13; H 2.84; N 9.76.

20
N. TROTSKO ET AL.
2-Methoxy-4-{[2-(phenylcarbamothioyl)hydrazinylidene]methyl}-
of incubation at 37 ꢀC – the cells are then in the log (logarithmic)
phase, during a period of active proliferation, but still have an
unoccupied space to divide. Afterwards, the culture medium was
discarded and replaced with 100 ml of different concentrations of
new tested compounds and reference antiproliferative agent: irino-
tecan. The stock solutions of the compounds were prepared in
DMSO, and solvent control was being tested concurrently to
exclude toxicity attributed to DMSO. Ten serial two-fold dilutions
using culture medium were performed to prepare different con-
centrations of the tested compounds, and the highest tested con-
centration was 200 mg/ml. Untreated cells (maintained in the
complete culture medium) served as a negative control. After
exposing cells for 48 h to the tested derivatives, the MTT assay
was performed as described earlier25. Three independent experi-
ments (n ¼ 3) were performed in a quadruplicate. The obtained
data were analysed using GraphPad Prism 5, version 5.03 software
(La Jolla, CA). The IC50 values were determined for tested com-
pounds revealing antiproliferative activity. Moreover, safety index
(SI) was calculated using the following formula:
phenyl (2,4-dioxo-1,3-thiazolidin-5-yl)acetate (26). Yield 88%, m.p.
1
182–184 ꢀC, H NMR d ppm (DMSO-d6): 3.40–3.43 m (2H, CH–CH2);
3.85 s (3H, OCH3); 4.83–4.87 m (1H, CH–CH2); 7.15–7.25 m,
7.36–7.47 m, 7.54–7.58 m, 7.67 d (8H, Ar, J ¼ 1.8 Hz); 8.14 s (1H,
CH¼N); 10.12 s, 11.90 s (NHCSNH); 12.13 s (1H, NH, thiazolidine),
13 C NMR d ppm (DMSO-d6): 36.1 (CH2CH), 46.9 (CH2CH), 56.7
(OCH3), 111.5, 121.4, 123.3, 126.0, 126.7, 128.6, 133.7, 139.6, 140.8,
142.6 (12 ꢁ Car.), 151.4 (CH¼N), 168.8 (C¼O), 172.7, 175.8
(2 ꢁ C¼O, thiazolidine), 176.6 (C¼S). MS (APCI) [M]þ m/
z ¼ 458.0958. Anal. calc. for C20H18N4O5S2 (458.51) (%): C 52.39; H
3.96; N 12.22. Found: C 52.40; H 3.95; N 12.21.
4-{[2-(Phenylcarbamothioyl)hydrazinylidene]methyl}phenyl
dioxo-1,3-thiazolidin-5-ylidene)acetate (27). Yield 84%, m.p.
199–202 ꢀC, 1H NMR
ppm (DMSO-d6): 7.06 s (1H, CH¼);
(2,4-
d
7.19–7.41 m, 7.56 d (7H, Ar, J ¼ 8.1 Hz); 8.02 d (2H, 4-O–C6H4,
J ¼ 8.7 Hz); 8.18 s (1H, CH¼N); 10.16 s, 11.88 s (NHCSNH); 12.72 s
(1H, NH, thiazolidine), 13 C NMR d ppm (DMSO-d6): 116.6 (CH ¼ C),
122.3, 125.9, 126.5, 128.5, 129.4, 132.8, 139.5, 142.2 (12 ꢁ Car.),
145.6 (CH¼C), 151.6 (CH¼N), 164.2 (C¼O), 166.8, 169.5 (2 ꢁ C¼O,
thiazolidine), 176.6 (C¼S). MS (APCI), fragment ion (m/z): 353.0611;
333.9962; 275.0140; 155.9762; 136.0231; 94.0658. Anal. calc. for
C19H14N4O4S2 (426.47) (%): C 53.51; H 3.31; N 13.14. Found: C
53.47; H 3.25; N 13.11.
ð
Þ
SI ¼ IC50 normal cells BJ =IC50 cancer cells
Microbiological tests
Newly synthesised compounds’ antibacterial activities were tested
in vitro against six aerobic bacteria reference strains: three Gram-
positive (Staphylococcus aureus ATCC 25923, methicillin-resistant S.
aureus MIKROBANK 14.001 reference strain from National
Medicines Institute in Warsaw, and Staphylococcus epidermidis
ATCC 12228) and three Gram-negative (Escherichia coli ATCC
25922, Proteus mirabilis ATCC 12453, and Pseudomonas aeruginosa
ATCC 9027).
2-Methoxy-4-{[2-(phenylcarbamothioyl)hydrazinylidene]methyl}-
phenyl (2,4-dioxo-1,3-thiazolidin-5-ylidene)acetate (28). Yield 93%,
m.p. 186–187 ꢀC, 1H NMR d ppm (DMSO-d6): 3.86 s (3H, OCH3);
7.08 s (1H, CH¼); 7.20–7.28 m, 7.39 t, 7.48–7.58 m, 7.71 d (8H, Ar,
J1¼7.5 Hz, J2¼1.5 Hz); 8.17 s (1H, CH¼N); 10.14 s, 11.92 s (NHCSNH);
12.94 s (1H, NH, thiazolidine), 13 C NMR d ppm (DMSO-d6): 56.7
(OCH3), 115.9 (CH ¼ C), 111.5, 121.4, 123.4, 126.0, 126.7, 128.6,
134.1, 139.6, 140.5, 142.5 (12 ꢁ Car.), 146.0 (CH¼C), 151.4 (CH¼N),
163.7(C¼O), 166.5, 169.4 (2 ꢁ C¼O, thiazolidine), 176.6 (C¼S). MS
(APCI), fragment ion (m/z): 383.0639; 302.0948; 150.0544; 136.0216;
94.0655. Anal. calc. for C20H16N4O5S2 (456.49) (%): C 52.62; H 3.53;
N 12.27. Found: C 52.63; H 3.54; N 12.30.
Bacterial cell suspensions equivalent to 0.5 McFarland (1.5 ꢁ 108
CFU/ml) were prepared in 0.85% sterile saline. Tested compounds’
stock solutions (50 mg/ml) were dissolved in dimethyl sulphoxide
(DMSO). DMSO final concentration did not inhibit bacteria growth.
Bacteria strain susceptibility to tested agents was determined
by broth microdilution method. Broth microdilution method was
carried out in accordance with Clinical and Laboratory Standards
Institute recommendations and was used as reference method26.
Mueller–Hinton broth containing two-fold dilutions of the tested
substances (at concentrations ranging from 3.91 to 1000 mg/ml)
was used to determine their minimal inhibitory concentration
(MIC) which is the lowest concentration of an antibacterial that
will inhibit the visible growth of a microorganism after 18 h of
incubation). Mueller–Hinton broth is recommended as the medium
of choice for susceptibility testing of commonly isolated, rapidly
growing aerobic, or facultative organisms. Gentamicin (KRKA,
Slovenia), an aminoglycoside antibiotic active against a wide range
of human bacterial infections, mostly Gram-negative bacteria
including Pseudomonas, Proteus, Serratia, and the Gram-positive
Staphylococcus, was used as a control antibacterial agent (at con-
centration from 0.12 mg to 1000 mg).
Bioassays
Antiproliferative activity in vitro
The experiment was carried out using normal human skin fibro-
blasts (BJ) and tumour cell lines: human lung carcinoma (A549),
human hepatocellular carcinoma (HepG2), and human breast
adenocarcinoma (MCF-7). All cell lines were obtained from ATCC
(American Type Culture Collection, England, UK) and cultured in
the appropriate, recommended by ATCC, culture medium, add-
itionally supplemented with 10% foetal bovine serum and antibi-
otics: 100 U/ml penicillin and 100 mg/ml streptomycin. The cells
were maintained at 37 ꢀC in a humidified atmosphere of 5% CO2
and 95% air. Antiproliferative activity of the new compounds was
assessed by, commonly used for this purpose, colorimetric MTT
assay. Cells with active mitochondrial dehydrogenases reduced
intracellularly the tetrazolium dye into purple formazan, which is
proportional to the number of viable cells. The cells were seeded
in 96-multiwell plates in 100 ll of the appropriate complete cul-
ture medium at a concentration of 7 ꢁ 103 cells/well in the case of
BJ cells (normal fibroblasts), 1 ꢁ 104 cells/well in the case of A549
cells (lung cancer), 1.5 ꢁ 104 cells/well in the case of HepG2 cells
Tested compounds two-fold dilutions in Mueller–Hinton broth
without bacteria were blank control wells (incubated on equal
conditions). Polystyrene trays containing 96 wells were filled with
serial two-fold Mueller–Hinton broth dilutions of tested agents.
Next, the broth was inoculated with 20 ll of 0.5 McFarland micro-
bial suspension (previously diluted 1:10 by Mueller–Hinton broth).
Total volume per each well was 200 ll. After incubation (at 35 ꢀC
for 18 h), bacterial cultures’ optical density was measured by
(liver cancer), and 2 ꢁ 104 cells/well in the case of MCF-7 cells microplate reader (BioTek Synergy H4 Hybrid Reader) at 600 nm.
(breast cancer) so as to obtain 60–70% confluent culture after 24 h The MIC values were determined by the comparison of broth

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
21
bacterial culture OD600 containing tested concentrations of exam- were prepared from acetic acid chlorides3,4 and a series of
hydroxybenzaldehydes (namely salicylaldehyde, 3-hydroxybenzal-
dehyde, 4-hydroxybenzaldehyde, and vanillin). The reaction was
provided in anhydrous 1,4-dioxane medium in the presence of
anhydrous pyridine by previously described method4,29. Then,
compounds 5–11 reacted with benzhydrazide derivatives (3-
chlorobenzhydrazide and 2,4-dichlorobenzhydrazide), and thiose-
micarbazide or 4-substituted-3-thiosemicarbazide derivatives
(4-phenyl-3-thiosemicarbazide and 4-(4-methylphenyl)-3-thiosemi-
carbazide) in the presence of anhydrous ethanol. The result of
this reaction was obtaining target compounds: hydrazones12–21
ined substances to blank control wells.
Results and discussions
Chemistry
2-(2,4-Dioxothiazolidin-5-yl)acetic chloride3 and 2-(2,4-dioxothiazo-
lidin-5-ylidene)acetic chloride4 were used as a starting material for
the synthesis of target compounds. Those acetic chlorides3,4 were
synthesised by the reaction of 2-(2,4-dioxothiazolidin-5-yl)acetic
acid or 2-(2,4-dioxothiazolidin-5-ylidene)acetic acid with thionyl
chloride in anhydrous 1,4-dioxane medium. In turn, 2-(2,4-dioxo-
thiazolidin-5-yl)acetic acid1 was prepared from thiourea and maleic
and thiosemicarbazones22–28
.
The synthesis of these com-
pounds12–28 was achieved through an efficient synthetic route
outlined in Figure 2.
anhydride in the presence of concentrated hydrochloric acid27
.
The structure of the new derivatives was confirmed by an
elemental analysis, 1H NMR and 13 C NMR and MS spectra. In 1H
NMR spectra, the protons of CH¼N group of all the newly synthes-
ised compounds showed a singlet signal at d ꢂ 8.14–8.51 ppm. The
protons of NHCSNH group resonated as two singlets in the range
of 10.10–10.17 ppm and 11.84–11.92 ppm correspondingly. The
2-(2,4-dioxothiazolidin-5-ylidene)acetic acid2 was obtained by the
reaction of compound 1 with bromine in the presence of acetic
acid28. The reactions are shown in Figure 1.
The next step of the synthesis was obtaining corresponding
formylphenyl 2-(2,4-dioxothiazolidin-5-yl)acetate and formylphenyl
2-(2,4-dioxothiazolidin-5-ylidene)acetate5–11
.
Compounds 5–11 protons of NH group of the thiazolidine ring appeared in the
O
O
O
NH2
i
NH
ii
NH
+
O
S
2
S
NH2
HO
S
O
HO
O
O
1
O
O
iii
iii
O
O
NH
NH
S
Cl
O
S
Cl
O
O
4
O
3
Figure 1. Synthesis of 2-(2,4-dioxothiazolidin-5-yl)acetyl chloride and 2-(2,4-dioxothiazolidin-5-ylidene)acetyl chloride. Reagent and conditions: (i) HCl, reflux; (ii) Br2,
CH3COOH, reflux; (iii) SOCl2, DMF, 1,4-dioxane, reflux 1 h.
O
O
OHC
i
NH
CHO
Z
NH
+
Z
S
HO
Cl
O
S
O
R
O
O
R
3, 4
O
5-11
ii
iii
H
O
S
N
H
H
N
O
N
N
NH
R1
R2
N
NH
Z
O
Z
S
O
R
O
S
O
R
O
O
12-21
O
22-28
R=H, OCH3, R1=3-Cl, 2,4-diCl, R2=H, C6H5, 4-CH3-C6H4
Z - single or double bond
Figure 2. Synthesis of target compounds (12–28) 2-(2,4-dioxothiazolidin-5-yl)acetic and 2-(2,4-dioxothiazolidin-5-ylidene)acetic acid derivatives. Reagent and conditions:
(i) pyridine, 1,4-dioxane, rt, after 2 h acidified of solution of hydrochloric acid; (ii) 3-chlorobenzhydrazide or 2,4-dichlorobenzhydrazide, anhydrous ethanol, reflux; (iii) cor-
responding 4-substituted thiosemicarbazide derivatives, anhydrous ethanol, reflux.

22
N. TROTSKO ET AL.
12.13–12.16 ppm region for compounds (12–17 and 22–26) and revealed the highest antiproliferative activity against human breast
12.72–12.98 ppm region for compounds as singlets18–21,27,28
adenocarcinoma cells (MCF-7). In the case of MCF-7 cells, the IC50
.
The presence of all carbon atoms for compounds12–28 was con- value of compound 18 was 13-fold lower (1.59 mg/ml) than IC50
firmed by 13 C NMR spectra. For compounds 12–17 and 22–26 value of irinotecan (21.08 mg/ml), whereas the SI value of com-
that are the derivatives of 2-(2,4-dioxothiazolidin-5-yl)acetic acid, pound 18 was three-fold higher when compared to the reference
carbon signal of two C¼O group of the thiazolidine ring appeared substance. In the case of human hepatocellular carcinoma (HepG2)
in the 172.7 and 175.8–176.0 ppm regions. Signals of two C¼O and human lung carcinoma (A549) cells, the IC50 values of com-
group of the thiazolidine ring for the 2-(2,4-dioxothiazolidin-5-ylili- pound 18 were also very low: 3.86 mg/ml (four-fold lower com-
dene)acetic acid derivatives 18–21, 27, and 28 were visible at dꢂ pared to irinotecan) and 8.08 mg/ml (five-fold lower compared to
166.5–166.8 ppm and 169.3–169.6 ppm ranges, respectively. The irinotecan), respectively. Comparing the activity of compound 18
detailed results of 1H NMR and 13 C NMR spectra and MS are pre-
to the activity of structurally similar 2,4-dioxothiazolidine-5-acetic
acid derivatives synthesised by Alegaon et al.6, it was observed
sented in the experimental part.
Molecular weight of compounds 12–21, 23, 24, and 26 were that compound 18 showed higher activity against MCF-7 and
confirmed by the mass spectra. In cases of compounds 22, 25, 27, A549 tumour cells. In their research, IC50 of tumour cell lines MCF-
and 28, a molecular ion was not observed in the mass spectra. 7 and A549 for the most active derivative 2-(2,4-dioxo-1,3-thiazoli-
The fragmentation was presented in the experimental part and din-5-yl)-N-[5-(3,4,5-trimethoxyphenyl)-1,3,4-thiadiazol-2-yl]aceta-
mide was 15.28 and 12.7 mg/ml, respectively.
ways of fragmentation were depicted in the supplementary
material.
The derivatives of 3-formylphenyl 2-(2,4-dioxo-1,3-thiazolidin-5-
yl)acetate with the 4-phenyl-3-thiosemicarbazide (compound 22)
and 4-(4-methylphenyl)-3-thiosemicarbazide structure (compound
23) also exhibited antiproliferative activity against all three tumour
cell lines (A549, HepG2, and MCF-7), but only IC50 of compound
23, determined using A549 cells (lung cancer), was lower com-
pared to the reference substance. However, the derivative 23
showed antiproliferative activity against liver cancer cells (HepG2)
comparable to irinotecan and with relatively high SI value (1.66),
indicating that this new compound has a great potential to be
used as an anticancer drug. The IC50 values of compound 22,
determined using all three cell lines, were relatively high.
Nevertheless, the SI values of compound 22 were >1, indicating
its high safety.
Biology
Antiproliferative evaluation
Among all the new compounds tested, only 16, 18, 20, 21, 22,
and 23 derivatives revealed antiproliferative activity against
tumour cell lines (Table 1). Compound 16 exhibited slight anti-
cancer activity only against human breast adenocarcinoma cells
(MCF-7). However, IC50 (concentration causing inhibition of cell
proliferation by 50%) of compound 16 determined using normal
human skin fibroblasts (BJ) was higher than IC50 of compound 16
estimated using MCF-7 cells, and the SI was relatively high and
>1. Note that the higher SI (SI > 1) is, the safer compound is (less
cytotoxic against normal cells). Also, agents revealing SI < 1 are
considered unsafe and highly cytotoxic. The slight modification of
the structure of compound 16 (changing a single bond in the pos-
ition 5 of the thiazolidine-2,4-dione to the double bond) resulted
in a new compound 21 that revealed broader antiproliferative
activity, also against human hepatocellular carcinoma cells
(HepG2). However, the SI value of compound 21 was low, indicat-
ing its high cytotoxicity against normal cells. Compound 20, an
analogue of the derivative 21 characterised by the lack of a
methoxy group in a 4-formylphenyl, showed antiproliferative activ-
ity only against HepG2 cells. However, similar to compound 21,
the derivative 20 revealed low SI value. Compound 18, character-
ised by the change in the configuration of formylphenyl (from
4-formyl-phenyl to 2-formyl-phenyl), appeared to be the most
promising antiproliferative agent among all tested compounds.
Compound 18 exhibited high antiproliferative activity against all
tumour cell lines used in the experiment (lung, breast, and liver
cancer) and high SI values (>1). Moreover, the derivative 18
showed meaningfully lower IC50 values when compared to the ref-
erence agent, irinotecan, which is an inhibitor of topoisomerase I
and is commonly used in cancer treatment. Compound 18
Antibacterial evaluation
Antibacterial activity of the newly synthesised compounds12–28 in
vitro was screened for the bacteria growth inhibition using broth
microdilution method (Table 2). Reference strains of Gram-nega-
tive and Gram-positive bacterial species were used. According to
Table 2. Newly synthesised compounds’ influence on the Gram-positive bacterial
strains growth – MIC (lg/ml) values.
Minimal inhibitory concentration MIC (lg/ml)
Staphylococcus
Staphylococcus aureus
Staphylococcus
Compound
aureus 25923
MIKROBANK 14.001
epidermidis 12228
12
13
15
17
18
23
25
27
28
500
500
500
250
62.5
125
250
125
125
500
500
500
250
125
125
250
125
125
500
500
500
500
125
62.5
250
125
125
Table 1. Antiproliferative activity of novel thiazolidine-2,4-dione derivatives and irinotecan used as a reference substance.
A549
HepG2
MCF-7
BJ
Cell line compound
IC50 [mg/ml] SEM SI [IC50BJ/IC50
]
IC50 [mg/ml] SEM SI [IC50BJ/IC50
]
IC50 [mg/ml] SEM SI [IC50BJ/IC50
]
IC50[mg/ml] SEM
16
18
20
21
22
23
117.2 1.23
1.59 1.06
1.39
6.32
163.3 1.34
10.05 1.18
11.35 1.05
31.42 1.08
81.62 1.11
25.16 1.52
38.69 1.37
8.08 1.05
1.24
3.86 1.07
26.96 1.02
48.39 1.02
48.33 1.07
15.18 1.25
14.79 1.24
2.60
0.42
0.65
1.69
1.66
2.62
124 1.63
72.02 1.10
34.18 1.12
21.08 1.23
0.25
1.13
0.74
1.84
57.27 1.19
31.31 1.66
39.69 1.20
1.43
0.80
0.97
Irinotecan – reference drug

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
23
our screening results, some of the tested compounds showed
potential antibacterial activity against Gram-positive bacteria.
Compounds 12, 13, 15, 17, and 25 exhibited similar MIC values
for S. aureus ATCC 25923, methicillin-resistant S. aureus
MIKROBANK 14.001, and S. epidermidis ATCC 12228 strains. The
MIC values ranged from 250 mg/ml to 500 mg/ml. Compound 27
inhibited all the tested staphylococci reference strains in the con-
centration of 125 mg/ml. Compound 23 showed antibacterial activ-
ity in the concentration of 125 mg/ml for S. aureus ATCC 25923,
methicillin-resistant S. aureus MIKROBANK 14.001 strains and
exhibited better antibacterial activity against S. epidermidis ATCC
12228 strain (MIC value was 62.5 mg/ml). The same MIC value
(62.5 mg/ml) was estimated for compound 18 towards S. aureus
ATCC 25923, while MIC value for methicillin-resistant S. aureus
MIKROBANK 14.001 and S. epidermidis ATCC 12228 strains was
125 mg/ml which indicates a lower antibacterial activity of com-
pound 18 in comparison with the activity in case of S. aureus
ATCC 25923.
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of the modifications introduced to the thiazolidine-2,4-dione may
result in the synthesis of new compounds with a broad biological
activity spectrum inter alia anticancer and antibacterial properties.
Such outcome was observed especially in the case of the change
in the configuration of formylphenyl (from 4-formyl-phenyl to 2-
formyl-phenyl) in compound 18 and introduction of the 4-(4-
methylphenyl)-3-thiosemicarbazide structure into compound 23.
The above-mentioned compounds revealed anticancer as well as
antibacterial activity. However, they showed relatively high MIC
values; thus, their application just as antibacterial agents is limited.
However, it should be noted that compounds 18 and 23 proved
to possess very promising anticancer properties, so the extension
of their activity towards antibacterial properties may be an add-
itional advantage as cancer patients have a reduced immunity and
are particularly sensitive to bacterial infections.
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zolyl-2, 4-thiazolidinediones as antibacterial and antifungal
agents. Org Med Chem Lett 2011;1:15.
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Conclusion
Conducted research resulted in the synthesis of new thiazolidine-
2,4-dione derivatives exhibiting antiproliferative activity. Among all
tested derivatives, compound 18, characterised by the change in
the configuration of formylphenyl from 4-formylphenyl to 2-for-
mylphenyl, revealed the highest antiproliferative activity against
human lung, breast, and liver cancer cells. More importantly, the
derivative 18 showed meaningfully lower IC50 values when com-
pared to the reference substance, irinotecan, and relatively high SI
values. Moreover, compound 18 is characterised by antibacterial
activity against some Gram-positive and Gram-negative strains. It
is a very important quality as cancer patients have reduced
immunity and are particularly sensitive to all kinds of infections,
bacterial infections in particular. Therefore, it may be implied that
the novel compound 18 appears to be a very promising agent for
anticancer treatment.
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zones. Bioorg Med Chem Lett 2008;18:6248–50.
20. Patel DH, Divatia SM, Clercq E. Synthesis of some novel thio-
semicarbazone derivatives having anticancer, anti-HIV as
well as antibacterial activity. Indian J Chem 2013;44:535–45.
Disclosure statement
The authors declare that there are no conflicts of interest.

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