Design, synthesis and in vitro antiproliferative activity of new thiazolidinedione-1,3,4-oxadiazole hybrids as thymidylate synthase inhibitors

Article abstract of DOI:10.1080/14756366.2020.1759581

Thymidylate synthase (TS) has been an attention-grabbing area of research for the treatment of cancers due to their role in DNA biosynthesis. In the present study, we have synthesised a library of thiazolidinedione-1,3,4-oxadiazole hybrids as TS inhibitors. All the synthesised hybrids followed Lipinski and Veber rules which indicated good drug likeness properties upon oral administration. Among the synthesised hybrids, compound 9 and 10 displayed 4.5 and 4.4 folds activity of 5-Fluorouracil, respectively against MCF-7 cell line whereas 3.1 and 2.5 folds cytotoxicity against HCT-116 cell line. Furthermore, compound 9 and 10 also inhibited TS enzyme with IC₅₀ = 1.67 and 2.21 μM, respectively. Finally, the docking studies of 9 and 10 were found to be consistent with in vitro TS results. From these studies, compound 9 and 10 has the potential to be developed as TS inhibitors.

Full text of DOI:10.1080/14756366.2020.1759581

Journal of Enzyme Inhibition and Medicinal Chemistry
ISSN: 1475-6366 (Print) 1475-6374 (Online) Journal homepage: https://www.tandfonline.com/loi/ienz20
Design, synthesis and in vitro antiproliferative
activity of new thiazolidinedione-1,3,4-oxadiazole
hybrids as thymidylate synthase inhibitors
Zohor Mohammad Mahdi Alzhrani, Mohammad Mahboob Alam, Thikryat
Neamatallah & Syed Nazreen
To cite this article: Zohor Mohammad Mahdi Alzhrani, Mohammad Mahboob Alam, Thikryat
Neamatallah & Syed Nazreen (2020) Design, synthesis and in vitro antiproliferative activity of new
thiazolidinedione-1,3,4-oxadiazole hybrids as thymidylate synthase inhibitors, Journal of Enzyme
Inhibition and Medicinal Chemistry, 35:1, 1116-1123
To link to this article: https://doi.org/10.1080/14756366.2020.1759581
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JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
2020, VOL. 35, NO. 1, 1116–1123
https://doi.org/10.1080/14756366.2020.1759581
BRIEF REPORT
Design, synthesis and in vitro antiproliferative activity of new thiazolidinedione-
1,3,4-oxadiazole hybrids as thymidylate synthase inhibitors
Zohor Mohammad Mahdi Alzhrania, Mohammad Mahboob Alama, Thikryat Neamatallahb and Syed Nazreena
b
aDepartment of Chemistry, Faculty of Science, Albaha University, Albaha, Saudi Arabia; Department of Pharmacology and Toxicology, Faculty
of Pharmacy, King Abdulaziz University, Jeddah, Saudi Arabia
ABSTRACT
ARTICLE HISTORY
Received 9 February 2020
Revised 11 April 2020
Accepted 18 April 2020
Thymidylate synthase (TS) has been an attention-grabbing area of research for the treatment of cancers
due to their role in DNA biosynthesis. In the present study, we have synthesised a library of thiazolidine-
dione-1,3,4-oxadiazole hybrids as TS inhibitors. All the synthesised hybrids followed Lipinski and Veber
rules which indicated good drug likeness properties upon oral administration. Among the synthesised
hybrids, compound 9 and 10 displayed 4.5 and 4.4 folds activity of 5-Fluorouracil, respectively against
MCF-7 cell line whereas 3.1 and 2.5 folds cytotoxicity against HCT-116 cell line. Furthermore, compound 9
and 10 also inhibited TS enzyme with IC50 ¼ 1.67 and 2.21 mM, respectively. Finally, the docking studies
of 9 and 10 were found to be consistent with in vitro TS results. From these studies, compound 9 and 10
has the potential to be developed as TS inhibitors.
KEYWORDS
2,4-thiazolidinediones; 1,3,4-
oxadiazoles; thymidylate
synthase; molecular
docking; pharmacokinetics
Introduction
medicinal chemistry due to its wide applications. Zibotentan
(ZD4054) containing 1,3,4-oxadiazole pharmacophore is an orally
available selective antagonist of the ET-A receptor with potential
antineoplastic effect for the treatment of various type of can-
cers.23,24 Recently, novel 1,3,4-oxadiazole bearing thioether deriva-
tives has been reported as potential TS inhibitors.25
Combination of the important bioactive pharmacophores under
one construct plays an important role in medicinal chemistry for
the development of biologically active molecules with novel
entity.26 To the best of our knowledge, anticancer effect of thiazo-
lidinedione derivatives as TS inhibitors has not been reported till
now. Therefore, we tried to conjugate thiazolidinedione and 1,3,4-
oxadiazole under one construct to develop potential TS inhibitors.
The present work describes the synthesis, in silico pharmacokinetic
studies, in vitro antiproliferative and TS inhibitory activities. The
docking studies have also been carried out for the most active
compounds to understand the possible underlying molecular
interactions.
Despite the availability of various chemotherapeutic agents, can-
cer is the second highest cause of deaths next to cardiovascular
diseases.1,2 Thymidylate synthase (TS) has become an area of
interest in cancer chemotherapy due to their important role in
DNA biosynthesis.3,4 TS is responsible for the formation of deoxy-
thymidine monophosphate (dTMP) from deoxyuridine monophos-
phate (dUMP) which is further phosphorylated to triphosphate
group (dTTP), a direct precursor for DNA synthesis.5–7 TS inhibition
causes inhibition of thymidylate biosynthesis which in turn causes
cessation of cell growth and proliferation.8,9 Research on TS has
been going on since many years, but still it is a challenge for
medicinal chemists to develop new, safe and effective chemother-
apeutic agents as TS inhibitor.
Thiazolidinediones (glitazones) are insulin sensitisers used for
type II diabetes treatment. They are high-affinity ligands of PPAR-
c, which alleviates insulin resistance and effectively improves
plasma glucose levels.10–11 Beside hypoglycaemic agents, thiazoli-
dinedione derivatives are reported as anti-inflammatory,12 anti-
microbial13 and anticancer agents.14–16 It has been mentioned
that compounds which activate PPAR-c might lead to differenti-
ation induction of cancer cells.17 For example, a TZD analogue,
efatutazone (CS-7017) is a potent PPAR-c full agonist as well as a
cancer differentiation-inducing agent.18 PPAR-c agonists are
reported to inhibit components of the insulin growth factor 1
(IGF1) pathway and modulate the activity of AMP-activated
protein kinase (AMPK) pathway to reduce cancer risk.19,20
Thiazolidinediones derivatives have exerted anticancer effects by
various mechanism of action viz. by inhibiting PI3K-a,21 blockade
Experimental
General
All the reagents and chemicals used in the present study were
procured from Sigma Aldrich (Germany) and Loba (India). IR was
recorded on Thermos scientific iS-50 by direct sampling method.
NMR analysis was performed on Bruker 300 and 850 MHz instru-
ments in either CDCl3 or DMSO-d6 solvents. Tetramethylsilane
(TMS) was used as internal reference. Chemical shift and coupling
constant are provided in Hertz and parts per million (ppm),
of the Raf/MEK/ERK and PI3K/Akt signalling pathways.22 On the respectively. Thermo scientific-LCQ Fleet (LCF10605) using electron
other hand, 1,3,4-oxadiazoles are promising candidates in spray ionisation method was used for recording the mass spectra
CONTACT Syed Nazreen
syed.nazreen@gmail.com
Department of Chemistry, Faculty of Science, Albaha University, Albaha, Saudi Arabia
Supplemental data for this article can be accessed here.
ß 2020 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,
distribution, and reproduction in any medium, provided the original work is properly cited.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
1117
and provided in m/z. Melting points were recorded on Stuart 50.86; H, 2.99; N, 8.90; O, 13.55; S, 6.79. Found: C, 50.84; H, 2.98; N,
SMP40. Elemental analyses were performed on LEECO Elementar 8.92; O, 13.57; S, 6.81.
(Z)-5-(4-methoxybenzylidene)-3-((5-p-tolyl-1,3,4-oxadiazol-2-
yl)methyl)thiazolidine-2,4-dione (11). Yield 72% mp
Elemental Analyser. The elemental analysis data were reported in
% standard and found to be within 0.4% of the calculated val-
ues. Purity of the compounds was checked on thin layer chroma-
tography using silica gel G plate (Merck Germany). The spectral
data and synthetic method of compounds 4–6 are provided in
the Supplementary Material.
197–198 ꢀC,1H NMR (300 MHz, DMSO-d6) d: 2.38 (s, 3H), 3.83 (s,
3H), 5.19 (s, 2H), 7.12 (d, 2H, J ¼ 8.7 Hz), 7.40 (d, 2H, J ¼ 7.8 Hz),
7.62 (d, 2H, J ¼ 8.7 Hz), 7.85 (d, 2H, J ¼ 8.1 Hz), 7.97 (s, 1H); 13C
NMR (75 MHz, DMSO-d6) d: 21.58, 36.21, 56.01, 115.50, 117.73,
120.77, 125.70, 127.01, 130.45, 132.91, 134.52, 142.81, 161.37,
161.86, 165.17, 165.32, 167.30; ESI þ ve MS (m/z): 408 (M þ H)þ.
Anal. Calc. for C21H17N3O4S: C, 61.90; H, 4.21; N, 10.31; O, 15.71; S,
7.87. Found: C, 61.89; H, 4.20; N, 10.29; O, 15.72; S, 7.85.
Chemistry
General procedure for the synthesis of thiazolidiene-2,4 dione-
(Z)-5-(4-methoxybenzylidene)-3-((5-(2-hydroxyphenyl)-1,3,4-
oxadiazol-2-yl)methyl)thiazol idine-2,4-dione (12). Yield 65%
mp 217–218 ꢀC, 1H NMR (850 MHz, CDCl3) d: 3.88 (s, 3H), 5.24 (s,
2H), 6.99–7.02 (m, 3H), 7.11 (d, 1H, J ¼ 8.5 Hz), 7.44–7.46 (m, 1H),
7.49 (d, 2H, J ¼ 8.5 Hz), 7.72 (dd, 1H, J ¼ 9.3, 1.7 Hz), 7.95 (s, 1H),
9.94 (s, 1H); 13C NMR (213 MHz, CDCl3) d: 35.55, 55.59, 107.60,
114.95, 117.11, 117.67, 120.01, 125.48, 126.76, 132.57, 134.09,
135.61, 157.63, 159.19, 161.92, 165.21, 165.29, 167.27; ESI þ ve MS
(m/z): 410 (M þ H)þ. Anal. Calc. for C20H15N3O5S: C, 58.67; H, 3.69;
N, 10.26; O, 19.54; S, 7.83. Found: C, 58.65; H, 3.68; N, 10.27; O,
19.55; S, 7.85.
(Z)-5-(4-methoxybenzylidene)-3-((5-o-tolyl-1,3,4-oxadiazol-2-
yl)methyl)thiazolidine-2,4-dione (13). Yield 57% mp 149–150 ꢀC,
1H NMR (300 MHz, DMSO-d6) d: 2.49 (s, 3H), 3.83 (s, 3H), 5.22 (s,
2H), 7.12 (d, 2H, J ¼ 8.7 Hz), 7.56–7.72 (m, 4H), 7.92–7.98 (m, 3H);
13C NMR (75 MHz, DMSO-d6) d: 21.55, 36.23, 56.01, 115.53, 117.72,
121.78, 125.79, 127.21, 128.38, 131.55, 132.94, 161.39, 161.90,
162.26, 165.38, 167.38; ESI þ ve MS (m/z): 408 (M þ H)þ. Anal. Calc.
for C21H17N3O4S: C, 61.90; H, 4.21; N, 10.31; O, 15.71; S, 7.87.
Found: C, 61.89; H, 4.20; N, 10.29; O, 15.72; S, 7.85.
(Z)-5-(4-methoxybenzylidene)-3-((5-(3-nitrophenyl)-1,3,4-
oxadiazol-2-yl)methyl)thiazolidin e-2,4-dione (14). Yield 75%
mp 178–179 ꢀC, 1H NMR (300 MHz, DMSO-d6) d: 3.84 (s, 3H), 5.24
(s, 2H), 7.12 (d, 2H, J ¼ 8.4 Hz), 7.63 (d, 2H, J ¼ 8.4 Hz), 7.88–8.07
(m, 2H), 8.34–8.64 (m, 2H), 8.75 (s, 1H); 13C NMR (213 MHz, DMSO-
d6) d: 36.16, 56.03, 114.87, 115.54, 117.77, 121.63, 124.98, 125.69,
127.07, 131.93, 132.96, 133.20, 134.54, 148.68, 161.87, 162.33,
163.69, 165.36, 167.38; ESI þ ve MS (m/z): 439 (M þ H)þ. Anal. Calc.
for C20H14N4O6S: C, 54.79; H, 3.22; N, 12.78; O, 21.90; S, 7.31.
Found: C, 54.80; H, 3.24; N, 12.75; O, 21.89; S, 7.32.
(Z)-5-(4-methoxybenzylidene)-3-((5-m-tolyl-1,3,4-oxadiazol-
2-yl)methyl)thiazolidine-2,4-dione (15). Yield 65% mp
184–185 ꢀC, 1H NMR (300 MHz, DMSO-d6) d: 2.40 (s, 3H), 3.84 (s,
3H), 5.13 (s, 2H), 7.13 (d, 2H, J ¼ 8.1 Hz), 7.38–7.51 (m, 2H),
7.62–7.69 (m, 2H), 7.76–7.96 (m, 2H), 7.98 (s, 1H); 13C NMR
(75 MHz, DMSO-d6) d: 21.27, 36.23, 56.02, 115.52, 117.75, 124.28,
125.71, 127.31, 129.85, 132.92, 133.31, 134.54, 139.44, 161.60,
161.87, 165.16, 167.35, 167.82; ESI þ ve MS (m/z): 408 (M þ H)þ.
Anal. Calc. for C21H17N3O4S: C, 61.90; H, 4.21; N, 10.31; O, 15.71; S,
7.87. Found: C, 61.89; H, 4.20; N, 10.29; O, 15.72; S, 7.85.
1,3,4-oxadiazole hybrids (7–21)
A mixture of compound 6 (0.01 mol) and different substituted aro-
matic hydrazides (0.01 mmol) in POCl3 (20 ml) was stirred and
refluxed for 10–12 h. After reaction completion, the reaction mix-
ture was poured onto crushed ice and neutralised with NaHCO3
solution. The resulting precipitate 7–21 was filtered, washed with
excess cold water and dried. Purification of compounds 7–21 was
done either by recrystallization in suitable solvents or by column
chromatography using n-hexane and ethylacetate as eluents. The
1H NMR, 13 C NMR and mass spectra of the compounds are pro-
vided in Supplementary Material.
(Z)-5-(4-methoxybenzylidene)-3-((5-phenyl-1,3,4-oxadiazol-
2-yl)methyl)thiazoli dine-2,4-dione (7). Yield 70% mp
212–216 ꢀC, 1H NMR (850 MHz, DMSO-d6) d: 3.84 (s, 3H), 4.48 (s,
2H), 7.13 (d, 2H, J ¼ 8.5 Hz), 7.44–7.47 (m, 2H), 7.61–7.73 (m, 5H),
7.96 (s, 1H); 13C NMR (213 MHz, DMSO-d6) d: 42.63, 56.03, 115.53,
117.60, 125.16, 125.64, 127.08, 129.10, 129.98, 132.98, 134.62,
161.53, 161.89, 165.53, 167.26, 167.46; ESI þ ve MS (m/z): 394
(M þ H)þ. Anal. Calc. for C20H15N3O4S: C, 61.06; H, 3.84; N, 10.68;
O, 16.27; S, 8.15. Found: C, 61.05; H, 3.85; N, 10.66; O, 16.29;
S, 8.14.
(Z)-5-(4-methoxybenzylidene)-3-((5-(3-chlorophenyl)-1,3,4-
oxadiazol-2-yl)methyl)thiazoli dine-2,4-dione (8). Yield 60%
mp 155–156 ꢀC, 1H NMR (850 MHz, DMSO-d6) d: 3.82 (s, 3H), 5.19
(s, 2H), 7.11 (d, 2H, J ¼ 9.3 Hz), 7.61–7.64 (m, 3H), 7.70–7.71 (m,
1H), 7.92–7.95 (m, 2H), 7.96 (s, 1H); 13C NMR (213 MHz, DMSO-d6)
d: 35.78, 55.63, 115.15, 117.38, 125.05, 125.31, 125.45, 126.17,
131.67, 132.13, 132.56, 134.11, 134.13, 161.47, 161.65, 163.62,
164.97, 166.99; ESI þ ve MS (m/z): 428 (M þ H)þ, 430 (M þ 2 þ H)þ.
Anal. Calc. for C20H14ClN3O4S: C, 56.14; H, 3.30; N, 9.82; O, 14.96; S,
7.49. Found: C, 56.11; H, 3.32; N, 9.83; O, 14.95; S, 7.50.
(Z)-5-(4-methoxybenzylidene)-3-((5-(2-chlorophenyl)-1,3,4-
oxadiazol-2-yl)methyl)thiazolid ine-2,4-dione (9). Yield 65%
mp 276–277, 1H NMR (300 MHz, DMSO-d6) d: 3.84 (s, 3H), 4.40 (s,
2H), 7.13 (d, 2H, J ¼ 8.7 Hz), 7.42–7.57 (m, 4H), 7.63 (d, 2H,
J ¼ 8.7 Hz), 7.95 (s, 1H); 13C NMR (75 MHz, DMSO-d6) d: 40.80,
56.01, 115.51, 118.19, 125.93, 127.59, 129.71, 130.29, 132.83,
134.02, 161.32, 161.78, 164.93, 165.68, 167.52; ESI þ ve MS (m/z):
428 (M þ H)þ, 430 (M þ 2 þ H)þ.Anal. Calc. for C20H14ClN3O4S: C,
56.14; H, 3.30; N, 9.82; O, 14.96; S, 7.49. Found: C, 56.11; H, 3.32; N,
9.83; O, 14.95; S, 7.50.
(Z)-5-(4-methoxybenzylidene)-3-((5-(phenoxymethyl)-1,3,4-
oxadiazol-2-yl)methyl)thiazolidine-2,4-dione (16). Yield 70%
mp 152–153 ꢀC, 1H NMR (300 MHz, DMSO-d6) d: 3.85 (s, 3H), 5.17
(s, 2H), 5.39 (s, 2H), 7.0–7.06 (m, 3H), 7.13 (d, 2H, J ¼ 9.0 Hz), 7.32
(t, 2H, J ¼ 6.9 Hz), 7.63 (d, 2H, J ¼ 8.7 Hz), 7.97 (s, 1H); 13C NMR
(75 MHz, DMSO-d6) d: 36.14, 56.02, 59.87, 115.34, 115.54, 117.58,
122.28, 125.67, 130.09, 132.94, 134.71, 157.70, 161.92, 162.44,
(Z)-5-(4-methoxybenzylidene)-3-((5-(4-bromophenyl)-1,3,4-
oxadiazol-2-yl)methyl)thiazolid ine-2,4-dione (10). Yield 65%
mp 263–264 ꢀC, 1H NMR (300 MHz, DMSO-d6) d: 3.83 (s, 3H), 4.41
(s, 2H), 7.12 (d, 2H, J ¼ 8.7 Hz), 7.63 (d, 2H, J ¼ 8.4 Hz), 7.71 (d, 2H,
J ¼ 8.7 Hz), 7.80 (d, 2H, J ¼ 8.4 Hz), 7.94 (s, 1H); 13C NMR (75 MHz,
DMSO-d6) d: 36.50, 56.01, 115.51, 120.59, 125.78, 129.98, 132.06, 164.17, 167.23; ESI þ ve MS (m/z): 424 (M þ H)þ. Anal. Calc. for
132.83, 155.22, 161.98, 165.76, 166.42, 166.76; ESI þ ve MS (m/z): C21H17N3O5S: C, 59.57; H, 4.05; N, 9.92; O, 18.89; S, 7.57. Found: C,
472 (M þ H)þ, 474 (M þ 2 þ H)þ.Anal. Calc. for C20H14BrN3O4S: C, 59.59; H, 4.04; N, 9.91; O, 18.90; S, 7.53.

1118
Z. M. M. ALZHRANI ET AL.
(Z)-3-((5-((3-chlorophenoxy)methyl)-1,3,4-oxadiazol-2-yl)
methyl)-5-(4-methoxybenzylidene)thiazolidine-2,4-dione (17).
Yield 72% mp 142–143 ꢀC, 1H NMR (400 MHz, CDCl3) d: 3.86 (s,
3H), 5.14 (s, 2H), 5.21 (s, 2H), 6.86–6.89 (m, 1H), 6.98–7.0 (m, 3H),
7.19–7.25 (m, 2H), 7.47 (d, 2H, J ¼ 8.8 Hz), 7.91 (s, 1H); 13C NMR
(100 MHz, CDCl3) d: 35.63, 55.56, 59.90, 113.08, 114.87, 114.95,
115.69, 122.62, 125.54, 130.53, 132.38, 132.52, 135.20, 135.49,
158.07, 161.93, 162.87, 165.23, 167.19; ESI þ ve MS (m/z): 458
(M þ H)þ, 460 (M þ 2 þ H)þ. Anal. Calc. for C21H16ClN3O5S: C, 55.08;
H, 3.52; N, 9.18; O, 17.47; S, 7.00. Found: C, 55.07; H, 3.54; N, 9.19;
O, 17.45; S, 7.01.
(Z)-3-((5-((2,3-dichlorophenoxy)methyl)-1,3,4-oxadiazol-2-yl)
methyl)-5-(4-methoxybenzylidene)thiazolidine-2,4-dione (18).
Yield 72% mp 209–210 ꢀC, 1H NMR (300 MHz, CDCl3) d: 3.87 (s,
3H), 5.15 (s, 2H), 5.31 (s, 2H), 6.98–7.01 (m, 3H), 7.12–7.15 (m, 2H),
7.48 (d, 2H, J ¼ 6.6 Hz), 7.91 (s, 1H); 13C NMR (100 MHz, CDCl3) d:
35.64, 55.54, 61.23, 112.90, 114.96, 124.42, 125.53, 127.54, 132.52,
135.50, 155.69, 161.94, 162.58, 167.15, 168.32; ESI þ ve MS (m/z):
492 (M þ H)þ, 494 (M þ 2 þ H)þ. Anal. Calc. for C21H15Cl2N3O5S: C,
51.23; H, 3.07; N, 8.53; O, 16.25; S, 6.51. Found: C, 51.21; H, 3.06; N,
8.51; O, 16.27; S, 6.50.
(Z)-5-(4-methoxybenzylidene)-3-((5-((naphthalen-1-yloxy)-
methyl)-1,3,4-oxadiazol-2-yl)methyl) thiazolidine-2,4-dione
(19). Yield 50% mp 220–221 ꢀC, 1H NMR (300 MHz, CDCl3) d: 3.86
(s, 3H), 5.15 (s, 2H), 5.43 (s, 2H), 6.93–7.01 (m, 3H), 7.33–7.51 (m,
7H), 7.76–7.79 (m, 1H), 7.90 (s, 1H); 13C NMR (213 MHz, CDCl3) d:
41.43, 55.52, 62.18, 115.12, 116.65, 117.90, 124.26, 125.59, 125.65,
128.12, 128.38, 132.40, 132.49, 134.80, 135.37, 161.52, 164.29,
165.56, 167.35, 167.19; ESI þ ve MS (m/z): 474 (M þ H)þ. Anal. Calc.
for C25H19N3O5S: C, 63.41; H, 4.04; N, 8.87; O, 16.89; S, 6.77. Found:
C, 63.42; H, 4.03; N, 8.90; O, 16.86; S, 6.78.
Molecular docking
Docking studies were performed at Intel(R) Core(TM) i3
CPU(2.3 GHz) with XP-based operating system (Windows 2007). 2 D
structures of the compounds were drawn by Marvin Sketch and
then converted into 3 D structures and saved in pdb file format.
Ligand preparation was done by assigning Gastegier charges,
merging non-polar hydrogen’s, and saving in PDBQT file format
using AutoDock Tools (ADT) 1.5.4. X-ray crystal structure of DNA
(PDB ID: 6QXG) was obtained from the Protein Data Bank (http://
www.rcsb.org/pdb). Gastegier charges were assigned to DNA and
saved in PDBQT file format using ADT. Preparation of parameter
files for grid and docking was done using ADT. Docking was per-
formed on AutoDock 4.0 (Scripps Research Institute, USA) consid-
ering all the rotatable bonds of the ligands as rotatable and DNA
as rigid.29 The grid centre was established by centring the grid
box on whole DNA. Grid boxsize of 60 ꢁ 80 ꢁ 110 Å with 0.375 Å
spacing was used. Macromolecule docking was performed using
an empirical-free energy function and Lamarckian Genetic
Algorithm, with an initial population of 150 randomly placed indi-
viduals, a maximum number of 2,500,000 energy evaluations, a
mutation rate of 0.02, and cross-over rate of 0.80. Fifty independ-
ent docking runs were performed of each ligand and DNA–ligand
complex for lowest free energy of binding conformation from the
largest cluster, which was written and saved in PDBQT format. The
results are summarised in Table 4 and Figure 1.
Results and discussion
Chemistry
Thiazolidinediones 3 was prepared by refluxing thiourea and
chloroacetic acid in hydrochloric acid and water which on
Knovenagel condensation with anisaldehyde in ethanol and aque-
ous sodium hydroxide gives intermediate benzylidene thiazolidi-
nediones 4. Alkylation of intermediate 4 with ethyl chloroacetate
in the presence of anhydrous potassium carbonate and dry acet-
one followed by acidic hydrolysis using glacial acetic acid and
hydrochloric acid gives the key intermediate 6. The key intermedi-
ate 6 was then refluxed with different aromatic acid hydrazides
and substituted phenoxy acetic acid hydrazides in the presence of
phosphorus oxychloride to give the target compounds 7–15 and
16–21, respectively in moderate to good yield (Scheme 1). The
structures of the synthesised hybrids were confirmed by different
analytical techniques such as 1H NMR, 13C NMR, IR spectroscopy,
elemental analyser and mass spectrometry. Formation of the con-
densed product 4 was supported by the presence of two doublets
at d 7.09 (J ¼ 8.7 Hz) and d 7.56 (J ¼ 8.7 Hz) which was assigned to
four aromatic protons, a singlet at d 7.75 for olefinic protons
bridging phenyl and thiazolidinedione rings and d 11.92 for NH of
TZD ring in the 1H NMR. This structure was supported by 13 C
NMR which revealed the downfield signals at d 161.45 for olefinic
C¼C and two signals at d 167.88 and d 168.40 for carbonyl groups
of TZD ring. The NH and C¼O bands of compound 4 were
observed at 3215 cmꢂ1(broad) and 1728 and 1690 cmꢂ1, respect-
ively in the IR spectrum. The conversion of compound 4 to com-
pound 5 was observed by the disappearance of NH signal at d
11.92 and appearance of a singlet at d 4.47 for N–CH2, quartette
at d 4.17 (J ¼ 7.2, 14.1 Hz) for O–CH2 and a triplet at d 1.21
(Z)-5-(4-methoxybenzylidene)-3-((5-((naphthalen-3-yloxy)-
methyl)-1,3,4-oxadiazol-2-yl)met hyl) thiazolidine-2,4-dione
(20). Yield 58% mp 210–211 ꢀC, 1H NMR (400 MHz, CDCl3) d: 3.86
(s, 3H), 5.15 (s, 2H), 5.35 (s, 2H), 6.98–7.0 (m, 3H), 7.18–7.20 (m,
2H), 7.46–7.48 (m, 4H), 7.74–7.76 (m, 2H), 7.89 (s, 1H); 13C NMR
(213 MHz, CDCl3) d: 41.45, 55.55, 62.21, 114.92, 116.64, 117.70,
123.93, 125.45, 125.65, 128.12, 128.38, 132.44, 132.56, 134.89,
135.46, 155.51, 161.71, 165.12, 166.19, 167.23, 167.64; ESI þ ve MS
(m/z): 474 (M þ H)þ. Anal. Calc. for C25H19N3O5S: C, 63.41; H, 4.04;
N, 8.87; O, 16.89; S, 6.77. Found: C, 63.42; H, 4.03; N, 8.90; O, 16.86;
S, 6.78.
(Z)-5-(4-methoxybenzylidene)-3-((5-((quinolin-8-yloxy)methyl)-
1,3,4-oxadiazol-2-yl)methyl) thiazolidine-2,4-dione (21). Yield
70% mp 161–162 ꢀC, 1H NMR (300 MHz, DMSO-d6) d: 3.84 (s, 3H),
5.19 (s, 2H), 5.62 (s, 2H), 7.11–7.14 (m, 2H), 7.36 (d, 1H, J ¼ 7.8 Hz),
7.46–7.64 (m, 5H), 7.96 (s, 1H); 8.33 (d, 1H, J ¼ 9.3 Hz), 8.85–8.86
(m, 1H); 13C NMR (75 MHz, DMSO-d6) d: 35.69, 58.53, 60.48, 111.46,
115.04, 117.05, 121.07, 121.25, 125.80, 128.53, 129.42, 132.45,
134.23, 138.45, 138.87, 149.32, 152.58, 160.47, 162.44, 163.32,
163.73, 166.72; ESI þ ve MS (m/z): 475 (M þ H)þ. Anal. Calc. for
C24H18N4O5S: C, 60.75; H, 3.82; N, 11.81; O, 16.86; S, 6.76. Found:
C, 60.74; H, 3.81; N, 11.80; O, 16.87; S, 6.77.
Cytotoxicity activity
The methods are provided in Supplementary material.
1
(J ¼ 6.9 Hz) for terminal methyl protons of the ester in the H NMR
In vitro thymidylate synthase assay
whereas in 13C NMR these signals appeared at d 42.60, d 62.15
TS activity was performed according to the method as described and d 14.38 for N–CH2, O–CH2 and terminal CH3, respectively. The
by Wahab and Friedkin27 with a slight modification according to absence of quartet and triplet and presence of broad signal at d
Santi.28 (Refer to Supplementary material for the method.)
13.44 in the 1H NMR of key intermediate 6 supported its

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
1119
Figure 1. Molecular docking of the active compounds 9, 10 and 5FU against Thimidylate synthase (TS) protein 6QXG. A: Binding mode of 9, 10 and 5FU at TS binding
site 3D plot. B: Binding mode of 9, 10 and 5FU at TS binding site 2D plot. 5FU: -5-fluorouracil.
formation. This data was further supported by 13 C NMR exhibiting formation. Further structural confirmation of the target com-
one additional downfield signal for new carbonyl group of the pounds 7–21 was provided by the presence of absorption bands
acid and IR spectrum exhibited broad signal for hydroxyl proton in the range of 1509–1589 cmꢂ1 for C¼N stretching of oxadiazole
at 3014 cmꢂ1. Compound 6 was finally confirmed by the appear- ring in the IR spectra and the signals in the range of d
ance of molecular ion peak at 292 (M ꢂ H)þ in mass spectrometry. 159.19–166.19 for C¼N of oxadiazole carbons in the 13 C NMR
The formation of the target compounds 7–21 was confirmed by spectra of these compounds. The target compounds 16–21 exhib-
the presence of additional aromatic protons in the range of d ited additional signal as singlet in the range of d 5.21–5.62 for
1
6.99–8.86 in their H NMR spectra. The absence of carboxyl proton O–CH2 bridged methylene aromatic ring and 1,3,4-oxadiazole ring.
1
signal in the target compounds 7–21 which was present in the H Finally, confirmatory evidence was done by their mass spec-
NMR spectrum of the key intermediate 6 also support their tral data.

1120
Z. M. M. ALZHRANI ET AL.
O
S
S
O
2
Anisaldehyde
HCl,
H2O
O
Cl
NH
H2N
NH2
N
H
3
H3C
OH
EtOH, 40%NaOH
S
O
O
1
O
4
O
O
O
Ethylchloroacetate
dry acetone, K2CO3
Gl. AcOH
N
O
N
H3C
S
OH
H3C
S
O
O
HCl
O
O
O
5
6
O
NH2
O
N
H
N
R
H3C
S
O
N
O
Ar
O
N
O
7-15
6
POCl3
O
N
H3C
S
O
O
Ar
O
N
O
O
NH2
N
N
H
16-21
R1
Cl
HO
17
7
8
12
Cl
Cl
Cl
18
13
14
Ar
19
20
9
Cl
NO2
Br
15
16
10
11
N
21
Scheme 1. Synthesis of thiazolidinedione-1,3,4-oxadiazole hybrids.
Pharmacokinetics studies/absorption, distribution, metabolism
elimination (ADME) predictions
should not be more than 5. Violation of any of these criteria
would result in problems of bioavailability upon oral administra-
tion. According to Veber et al.,31 number of rotatable bonds
should be ꢃ10 which is an indicator for good bioavailability. In
the present study, we calculated several parameters for predicting
drug-likeness properties of synthesised compounds. The thiazolidi-
nedione linked 1,3,4-oxadiazole hybrids (7–21) were subjected to
following in silico physicochemical studies: number of rotatable
bonds (nROTB), hydrogen bond acceptor (HBA), hydrogen bond
donor (HBD), lipophilicity (iLogP) and topological polar surface
area (TPSA). In silico %age absorption were calculated using the
reported formula [(%ABS ¼ 109–(0.345 X TPSA)]. From the results,
The capability of a drug to exhibit a pharmacological or thera-
peutic effect depends upon various physiochemical properties of
the drug. Nowadays, ADME predictions of molecules are studied
in initial phases of drug development by in silico approaches to
generate potential lead molecules. Drug-likeness in drug develop-
ment gives an idea whether the particular synthesised compound
is comparable to already existing drug molecules in the market.
The synthesised molecule expected to be an orally active drug
should obey Lipinski rule30 which states the following four criteria:
molecular weight should not be more than 500, hydrogen bond
acceptor should not be more than 10, hydrogen bond donor it was observed that % absorption was found to be in the range
should not be more than 5 and partition coefficient (Clog P) of 54.98–70.76%. Compound 7, 8, 9, 10, 11, and 13 showed

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
1121
Table 2. The IC50 (mM) of the synthesised compounds (7–21)
Table 1. Pharmacokinetics/ADME predictions of the target compounds 7–21.
against tested human cancer cell line (MCF-7 and HCT-116)a.
Lipinski parameters
Compound
MCF-7b
HCT-116c
No. MWa HBAb HBDc iLogPd Violations nROTBe TPSAf % ABSg BBBh GI ABSi
7
8
9
53.50
56.0
7.74
57.34
56.76
12.84
16.14
68.13
58.64
58.64
42.21
39.60
67.10
55.5
>100
69.50
>100
63.15
40.51
7
8
9
394
430
430
6
6
6
6
6
7
6
8
6
7
7
7
7
7
8
0
0
0
0
0
1
0
0
0
0
0
0
0
0
0
3.3
3.62
3.5
3.69
3.6
3.21
3.46
2.87
3.79
3.39
3.81
3.17
3.74
3.84
3.22
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
5
5
5
5
5
5
5
6
5
7
7
7
7
7
7
110.83 70.76 No High
110.83 70.76 No High
110.83 70.76 No High
110.83 70.76 No High
110.83 70.76 No High
131.06 63.78 No High
110.83 70.76 No High
10
11
12
13
14
15
16
17
18
19
20
21
5-FUd
7.87
10 474
11 408
12 410
13 408
14 439
15 408
16 424
17 460
18 494
19 474
20 474
21 475
58.37
56.28
56.3
43.37
47.8
156.65 54.98 No
Low
110.83 70.76 No High
120.06 67.60 No High
120.06 67.60 No High
120.06 67.60 No High
120.06 67.60 No High
120.06 67.60 No High
58.4
65.17
>100
61.6
>100
61.95
34.82
132.95 63.13 No
Low
aMolecular weight.
aIC50 values are the concentrations that cause 50% inhibition of
cancer cell growth. Data represent the mean values standard
deviation of three independent experiments performed in triplicate.
bBreast cancer (MCF-7); colorectal cancer (HCT-116).
bHydrogen bond acceptor.
cHydrogen bond donor.
d
Partition coefficient.
eNumber of rotatable bonds.
fTopological polar surface area.
gAbsorption %.
c5-FU: 5-florouracil was used as
tive control).
a reference drug (posi-
Bold values: significant cytotoxicity compared to standard drug
5-FU.
hBlood brain barrier.
iGastro-intestinal absorption.
cytotoxicity was observed by compounds 7, 8, 11, 12, 13, 16, 17,
19 and 21 with IC50 in the range 43.37–65.17 mM and
55.5–69.50 mM against MCF-7 and HCT-116, respectively while
compound 18 and 20 were found to be inactive (IC50 > 100)
against the tested cell lines. Among the synthesised compounds,
compound 9 and 10 were the most potent against both MCF-7
and HCT-116 cell line. From the cytotoxicity results, it was noted
that the 1,3,4-oxadiazole-thiazolidienedione hybrids derived from
substituted aromatic acids hydrazides (7–15) were found to
exhibit significant cytotoxicity (IC50 < 50 mM) than 1,3,4-oxadiazole-
thiazolidienedione hybrids obtained from substituted phenoxy
acetic acids hydrazides (16–21) against both the tested cell line.
Among the synthesised hybrids (7–15), compound 9 and 10 bear-
ing halogen at ortho and para position (o-Cl, p-Br) showed highest
cytotoxicity.
highest in silico % absorption 70.76% whereas compound 14
revealed 54.98 in silico % absorption. All the synthesised com-
pounds follow Lipinski rule of 5. Molecular weight ranges from
394 to 492 (<500), HBA range 6–8 (ꢃ10), HBD range 0–1 (ꢃ5),
iLogP (lipophilicity) was found to be in the range 2.87–3.84 (ꢃ5)
which suggest that these compounds upon administration possess
good drug likeness properties (Table 1). All the compounds except
14 and 21 exhibited high gastro-intestinal absorption and they
are impermeable to brain. Furthermore, all the compounds
showed nROTB in the range 5–7 (<10) suggesting good bioavail-
ability. All the tested compounds except 14 revealed TPSA range
110.83–132.95 Å2 (<140 Å2), which indicates good intestinal
absorption. From these parameters, these synthesised compounds
exhibited good drug likeness properties.
Biological evaluation
In vitro thymidylate synthase activity
Cytotoxicity assay
Compounds (9, 10, 14 and 15) which showed better cytotoxicity
in MTT assay were further tested for their in vitro TS activity to
confirm its mechanism of action. The TS inhibition results are pre-
sented in Table 3. Interestingly, these compounds exhibited
remarkable inhibition on the TS enzyme. Compound 9 and 10
revealed TS inhibition with IC50 ¼ 1.67 and 2.21 mM, respectively
compared to Pemetrexed (IC50 ¼ 8.2 mM,). The results of TS inhib-
ition were found to be in agreement with the cytotoxicity results.
The newly synthesised compounds 7–21 were tested for their
cytotoxicity against human cancer cell line viz. breast and colorec-
tal using MTT assay. The IC50 value of the synthesised compounds
is presented in Table 2. From the results, it has been observed
that the synthesised compounds showed significant to moderate
growth inhibition compared to the reference drug, 5-fluorouracil
(5FU). The cytotoxicity results revealed that compound 9, 10 and
15 showed promising activity. Compound 9 and 10 showed sig-
nificant cytotoxicity with IC50 ¼ 7.47 mM and IC50 ¼ 7.87 mM,
respectively against MCF-7 whereas reference drug 5-FU exhibited
Molecular docking studies
IC50 ¼ 34.82 mM. Compound 9 (IC50 ¼ 7.47 mM) and 10 (IC50
¼
Molecular docking is an intriguing technique to delineate the
mechanism of drug-receptor binding in a time and cost-effective
way. Among a number of software’s, AutoDock is known for
fast speed, easy availability, and high accuracy (rmsd ¼ 2 Å)
results.32–33 To support our human TS inhibition results and to get
an idea about the binding modes, we carried out shape-based
molecular docking studies of compounds 9 and 10 against TS pro-
tein (PDB ¼ 6QXG) using Autodock tools. The results of the find-
7.87 mM) displayed 4.5 and 4.4 folds activity of the 5-FU, respect-
ively against MCF-7 cell line. Against HCT-116 cell line, four com-
pounds 9, 10, 14 and 15 showed excellent cytotoxicity with IC50
value of 12.84, 16.14, 42.21 and 39.60 mM, respectively whereas
reference drug 5-FU exhibited IC50 ¼ 40.51 mM. It was noticed that
the same compound 9 and 10 showed 3.1 and 2.5 folds cytotox-
icity compared to5-FU against HCT-116 while compound 14 and
15 was found to be equipotent to the reference drug with IC50
42.2 mM and IC50
¼
¼
39.60 mM, respectively. The moderate ings are depicted in Figure 1 and Table 4.

1122
Z. M. M. ALZHRANI ET AL.
Table 3. In vitro thymidylate synthase (TS) activity of
the active compounds 9, 10, 14, 15 and PTX.
Acknowledgement
The authors thanks Chemistry department, Albaha university for
providing the necessary facilities to carry out this work.
Compounds
IC50 (mM)
9
1.67
2.21
9.4
7.9
8.2
10
14
15
PTX
Disclosure statement
No potential conflict of interest was reported by the author(s).
IC50 values are the mean SD of three separate
experiments. PTX: Pemetrexed.
References
Table 4. Docking scores of active compounds 9 and 10 against human thymidy-
late synthase protein 6QXG.
1. Goud NS, Ghouse SM, Arifuddin M, et al. Synthesis and bio-
logical evaluation of coumarin-1,3,4-oxadiazole hybrids as
selective carbonic anhydrase IX and XII inhibitors. Bioorg
Chem 2019;87:765–72.
2. Patel K, Karthikeyan C, Solomon VR, et al. Synthesis of some
coumarinyl chalcones and their antiproliferative activity
against breast cancer cell lines. Lett Drug Design Disc 2011;
8:308–11.
Compound
Docking score
Amino acid residue
9
10
5-FU
ꢂ9.09
ꢂ8.67
ꢂ4.22
Cys 195, Tyr 135 and Asn 226
Leu 221, Asn 226 and Tyr 230
Phe 80, Arg 78, Thy306 and Ile307
It has been reported that TS-inhibitors act by interacting with
the active sites of the receptor through multiple amino acid resi-
dues (viz. Arg50, Ser216, Asn 226, Asp218, His256, Tyr258, Arg50
and Arg215).34–35 Interestingly, our docking results are completely
corroborated the literature. We found that the compounds
showed good binding affinity towards TS protein (ꢂ9.09 and
ꢂ8.67 kcal/mol for 9 and 10, respectively) and interacted through
multiple H-bonds. Oxygen atom of 4-methoxy group was involved
in hydrogen bonding interaction with Cys 195, Tyr 135 and oxa-
diazole ring oxygen atom with Asn 226 in compound 9.Whereas
in compound 10, oxygen atom of methoxy group showed hydro-
gen bonding interaction with Tyr 230, thiazolidinedione C¼O
group at 4-position showed hydrogen bonding interaction with
Asn 226 and nitrogen atom of oxadiazole ring at 4-position with
Leu 221. Contrary to this, reference drug 5-fluorouracil (5-FU)
exhibited lower affinity (ꢂ4.22 kcal/mol) and interacted with Thy
306, Phe 80, Ile307 and Arg 78 via H-bonding interaction with
C¼O group at 2- and 4-position, N–H group at 1- and 3-position
respectively. Compounds 9 and 10 have different substituent in
the phenyl ring attached to the oxadiazole ring, bearing chlorine
at ortho position and bromine at para position, respectively but
due to flexibility of the molecule, compound 10 bend and interact
with amino residue Tyr 230, Asn 226 and Leu 221. Whereas, 5 flu-
orouracil is a small ligand in which all the electronegative atoms
interact with the amino acid residues except fluorine. These
results were in agreement with the in vitro results of compound 9
(IC50 ¼ 1.67) and 10 (IC50 ¼ 2.21 mM).
3. Lambratu R, Donna V, Monzur R, et al. Thymidylate synthase
as an oncogene: a novel role for an essential DNA synthesis
enzyme. Cancer Cell 2004;5:341–51.
4. Young DW. Studies on thymidylate synthase and dihydrofo-
late reductase-two enzymes involved in the synthesis of thy-
midine. Chem Soc Rev 1994; 23:119–28.
5. Kumar VP, Cisneros JA, Frey KM, et al. Structural studies pro-
vide clues for analog design of specific inhibitors of
Cryptosporidium hominis thymidylate synthase-dihydrofolate
reductase. Bioorg Med Chem Lett 2014;24:4158–61.
6. Catalano A, Luciani R, Carocci A, et al. X-ray crystal struc-
tures of Enterococcus faecalis thymidylate synthase with fol-
ate binding site inhibitors. Eur J Med Chem 2016;123:
649–64.
7. Onen FE, Boum Y, Jacquement C, et al. Design, synthesis
and evaluation of potent thymidylate synthase X inhibitors.
Bioorg Med Chem Lett 2008;18:3628–31.
8. Friedkin M, Kornberg A. The enzymatic conversion of deoxy-
uridylic acid to thymidylic acid and the participation of tet-
rahydrofolic acid. In: Mc Elroy WD and Glass B, eds. The
Chemical Basis of Heredity. Baltimore: Johns Hopkins Press;
1957:609–614.
9. Navalgund LG, Rossana C, Muench AJ, et al. Cell cycle regu-
lation of thymidylate synthetase gene expression in cultured
mouse fibroblasts. J Biol Chem 1980;255:7386–90.
10. Nazreen S, Alam MS, Hamid H, et al. Design, synthesis, in sil-
ico molecular docking and biological evaluation of novel
oxadiazole based thiazolidine-2,4-diones bis-heterocycles as
PPAR-c agonists. Eur J Med Chem 2014;87:175–85.
11. Unlusoy MC, Verspohl EJ, Ertan R. Synthesis and antidiabetic
activity of some new chromonyl-2,4-thiazolidinediones.
J EzyInhib Med Chem 2010; 25:784–9.
Conclusions
A library of synthesised thiazolidinedione-1,3,4-oxadiazole hybrids
have been screened for in vitro antiproliferative as well as TS activ-
ities. All the synthesised hybrids follow Lipinski and Veber rules
indicating good drug likeness properties upon oral administration.
Among the synthesised hybrids, compound 9 and 10 showed
excellent cytotoxicity against MCF-7 and HCT-116 cells. From the
in vitro TS activity, Compound 9 and 10 were found to be potent
inhibitors against TS protein with IC50 ¼ 1.67 and 2.21 mM,
respectively. The docking studies of compounds 9 and 10 were
found to be consistent with in vitro cytotoxicity and TS results.
From these studies, it can be concluded that compounds 9 and
10 have the potential to be developed as TS inhibitors.
12. Chen R, Yan J, Liu P, et al. Effects of thiazolidinedione ther-
apy on inflammatory markers of type 2 diabetes: a meta-
analysis of randomized controlled trials. PLoS ONE 2015;10:
e0123703.
13. Hu CF, Zhang PL, Sui YF, et al. Ethylenic conjugated couma-
rin thiazolidinediones as new efficient antimicrobial modula-
tors against clinical methicillin-resistant Staphylococcus
aureus. Bioorg Chem 2020; 94:103434.
14. Shankar S, Vuppu S. In vitro drug metabolism and pharma-
cokinetics of a novel thiazolidinedione derivative, a potential
anticancer compound. J Pharma Biomed Anal 2020;179:
113000.

JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
1123
15. Tokala R, Thatikonda S, Sana S, et al. Synthesis and in vitro
cytotoxicity evaluation of b-carboline-linked 2,4-thiazolidine-
anticancer/antimicrobial agents. Bioorg Med Chem 2013;21:
2286–97.
dione hybrids: potential DNA intercalation and apoptosis- 26. Abbot V, Sharma P, Dhiman S, et al. Small hybrid heteroaro-
inducing studies. New J Chem 2018;42:16226–36.
matics: resourceful biological tools in cancer research. RSC
16. Trotsko N, Przekora A, Zalewska Z, et al. Synthesis and
Adv 2017;7:28313–49.
in vitro antiproliferative and antibacterial activity of new 27. WahbaAJ FM. The enzymatic synthesis of thymidylate: i.
thiazolidine-2,4-dione derivatives. J EzyInhib Med Chem
early steps in the purification of thymidylate synthetase of
2018;33:17–24.
Escherichia coli. JBiol Chem 1962;237:3794.
17. Shimazaki N, Togashi N, Hanai M, et al. Anti-tumour activity 28. Davisson VJ, Sirawaraporn W, Santi DV. Expression of human
of CS-7017, a selective peroxisome proliferator-activated
receptor gamma agonist of thiazolidinedione class, in
thymidylate synthase in Escherichia coli. J Biol Chem 1989;
264:9145–8.
human tumour xenografts and a syngeneic tumour implant 29. Morris GM, Goodsell GS, Halliday RS, et al. Automated dock-
model. Eur J Cancer 2008;44:1734–43.
18. Smallridge RC, Copland JA, Brose MS, et al. Efatutazone, an
oral PPAR-c agonist, in combination with paclitaxel in ana-
ing using a lamarckian genetic algorithm and an empirical
binding free energy function. J Comput Chem 1998;19:
1639–62.
plastic thyroid cancer: results of a multicenter phase 1 trial. 30. Lipinski CA, Lombardo F, Dominy BW, et al. Experimental
J Clin Endocrin Metab 2013;98:2392–400.
and computational approaches to estimate solubility and
permeability in drug discovery and development settings.
Adv Drug Del Rev 2001;46:3–26.
19. Belfiore A, Genua M, Malaguarnera R. PPAR-c agonists and
their effects on IGF-i receptor signaling: implications for can-
cer. PPAR Res 2009;2009:1–18.
20. Lee WH, Kim SG. AMPK-dependent metabolic regulation by
PPAR agonists. PPAR Res 2010;2010:1–10.
31. Veber DF, Johnson SR, Cheng HY, et al. Molecular properties
that influence the oral bioavailability of drug candidates.
J Med Chem 2002;45:2615–23.
21. Knight SD, Adams ND, Burgess JL, et al. Discovery of 32. Patrick AH, Jonathan BC, John OT. Molecular docking of
GSK2126458, a highly potent inhibitor of pi3k and the mam-
intercalators and groove- binders to nucleic acids using
malian target of rapamycin. Med Chem Lett 2010;1:39–43.
autodock and surflex. J Chem Inf Model 2008;48:1602–15.
22. Liu K, Rao W, Parikh H, et al. 3,5-Disubstituted-thiazolidine- 33. Oleg T, Arthur JO. AutoDock Vina: improving the speed and
2,4-dione analogs as anticancer agents: design, synthesis
and biological characterization. Eur J Med Chem 2012;47:
125–36.
accuracy of docking with a new scoring function, efficient
optimization and multithreading. J Comput Chem 2010;31:
455–61.
23. James ND, Growcott JW. Zibotentan. Drugs Future 2009;34: 34. Lamia HTA, Taghreed ZS, Abeer MN. Design, synthesis, anti-
624–33.
cancer evaluation and docking studies of new pyrimidine
derivatives as potent thymidylate synthase inhibitors. Bioorg
Chem 2019;91:103159.
24. Shen W, Xi H, Li C, et al. Endothelin-A receptor in gastric
cancer and enhanced antitumor activity of trastuzumab in
combination with the endothelin—A receptor antagonist 35. Ahmed FE, Qasem MAA, Emad S. Design, synthesis, molecu-
ZD4054. Ann N Y Acad Sci 2019;1448:30–41.
25. Du QR, Li DD, Pi YZ, et al. Novel 1,3,4-oxadiazole thioether
derivatives targeting thymidylate synthase as dual
lar docking of new thiopyrimidine-5-carbonitrile derivatives
and their cytotoxic activity against HepG2 cell line. J Appl
Pharm Sci 2014;4:102–11.