Synthesis and anticancer activities of some new coumarin derivatives including the triazole ring and their in silico molecular docking studies

Article abstract of DOI:10.1002/ardp.202000284

The synthesis, docking study, and investigation of the anticancer activities of some coumarin derivatives containing the triazole ring are reported in this study. The newly synthesized compounds were screened for their in vitro anticancer activity against the cell lines CRL5807 (human bronchioalveolar carcinoma), CRL5826 (human squamous cell carcinoma), MDA-MB231 (human breast cancer cells), HTB177 (human lung cancer), PC-3 (human prostate adenocarcinoma), PANC-1 (human pancreatic cancer cells), used as cancer cells, and CCD34Lu (normal human lung fibroblasts), used as a healthy cell line. Cytotoxicity effects of the samples were determined by the MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay. In silico studies were also performed to explore the binding interactions of the molecules.

Full text of DOI:10.1002/ardp.202000284

Received: 4 August 2020
Revised: 21 September 2020
Accepted: 22 October 2020
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DOI: 10.1002/ardp.202000284
F U L L P A P E R
Synthesis and anticancer activities of some new coumarin
derivatives including the triazole ring and their in silico
molecular docking studies
Emre Menteşe1
| Adem Güner2 | Elifsu Polatlı3 | Mustafa Emirik1
Hakan Bektaş4 | Bahittin Kahveci5
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1Department of Chemistry, Faculty of Arts
and Sciences, Recep Tayyip Erdogan
University, Rize, Turkey
Abstract
The synthesis, docking study, and investigation of the anticancer activities of some
coumarin derivatives containing the triazole ring are reported in this study. The
newly synthesized compounds were screened for their in vitro anticancer activity
against the cell lines CRL5807 (human bronchioalveolar carcinoma), CRL5826
(human squamous cell carcinoma), MDA‐MB231 (human breast cancer cells),
HTB177 (human lung cancer), PC‐3 (human prostate adenocarcinoma), PANC‐1
(human pancreatic cancer cells), used as cancer cells, and CCD34Lu (normal human
lung fibroblasts), used as a healthy cell line. Cytotoxicity effects of the samples were
determined by the MTT (3‐(4,5‐dimethyl‐2‐thiazolyl)‐2,5‐diphenyl‐2H‐tetrazolium
bromide) assay. In silico studies were also performed to explore the binding
interactions of the molecules.
2Department of Biology, Faculty of Science
and Art, Giresun University, Giresun, Turkey
3Department of Bioengineering, Faculty of
Engineering, Ege University, İzmir, Turkey
4Department of Chemistry, Faculty of Science
and Art, Giresun University, Giresun, Turkey
5Department of Nutrition and Dietetics,
Faculty of Health Sciences, Karadeniz
Technical University, Trabzon, Turkey
Correspondence
Emre Menteşe, Department of Chemistry,
Faculty of Arts and Sciences, Recep Tayyip
Erdogan University, 53100 Rize, Turkey.
Email: emre.mentese@erdogan.edu.tr
K E Y W O R D S
anticancer, coumarin, cytotoxicity, molecular docking, triazole
1
INTRODUCTION
antifungal,[8–11] anti‐inflammatory,[12] anti‐HIV, and antihyperten-
sion.[13] Besides, they also act as α‐glucosidase and pancreatic lipase
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Cancer is a general definition of a broad group of diseases char-
acterized by the uncontrolled proliferation of abnormal cells and
metastasis to other organs of these cells. Moreover, cancer can af-
fect nearly any part of the body and has various cellular and mole-
cular subtypes, which can affect different molecular pathways.[1,2] It
is the second major reason for death worldwide, and it was predicted
to cause approximately 9.6 million deaths in 2018.[3] Recent reports
have shown that the most common types of cancer among men are
prostate, colorectal, lung, liver, and stomach cancer, whereas cervix,
lung, breast, colorectal, and thyroid cancer are the most widespread
cancer among women.[3]
inhibitors.[14–16] In addition to these, coumarins can be used as dyes
in the laser industry,[17] in the perfumery,[18] and as fluorescent laser
dyes.[19] The structure‐activity relationship in the previous studies
shows that the use of the coumarin ring in heterocyclic compounds
modulates inhibitory activities.[20,21] There are many new cancer
types resulting in a large number of deaths.[22] Biological investiga-
tions of coumarin derivatives showed the engrossment of innumer-
able pathways by which coumarins act as anticancer agents.[23]
Hybridization of coumarin and triazole is one of the most attractive
strategies for achieving new potent anticancer agents that have
better pharmacokinetic and pharmacodynamic properties.[24] Some
research works have demonstrated that coumarin–triazole hybrid
molecules have been investigated as potent anticancer agents
(Figure 1).[23–26] Recently, our research group has synthesized some
coumarin–triazole hybrid compounds as potent anticancer agents.[27]
Coumarins are very important compounds that widely occur in
natural products.[4] They are also named as chromones.[5] Natural
and synthetic derivatives of coumarins exhibit many biological
properties such as anticancer,[6] antimicrobial, antioxidant,[7]
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Arch Pharm. 2020;e2000284.
wileyonlinelibrary.com/journal/ardp © 2020 Deutsche Pharmazeutische Gesellschaft
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https://doi.org/10.1002/ardp.202000284

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FIGURE 1 Structures of recently reported coumarin–triazole derivatives with anticancer activity and the target compounds
SCHEME 1 The synthetic path of compounds 3a–f

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Cytotoxicity
Considering the abovementioned facts, the exploration of po-
2.2
tential and effective anticancer compounds is important for new
drug development in cancer research. In this context, a series of
novel coumarin derivatives containing triazole structures, which
exhibit very important biological properties such as antioxidant,
antimicrobial, antiviral, antitubercular, anti‐inflammatory, antic-
onvulsant, and anticancer activities,[28–31] was synthesized and their
anticancer activities were investigated.
According to test results (Table 1 and Figure S1), 3e and 3a showed
the lowest effect on all cell lines. 3a has no effect on PC‐3 cells. The
effect of 3e on CCD34Lu, HTB177, and PC‐3 cells was observed, but
IC50 could not be reached. Nevertheless, the most effective samples
were determined as 3b and 3d for all cells and their most affected
cells, respectively, MDA‐MB231 and CRL5807 cells. Compound 3a is
one of the less effective samples, with its IC50 value being more than
39 µg/ml. Compound 3b has the highest inhibitory effect on
MB‐MB231 cells, with its IC50 value being 0.86 0.04 µg/ml.
Molecule 3c also has the highest effect on MDA‐MB231 cells, with
its IC50 value being 4.10 0.4 µg/ml. Compound 3d has the highest
effect on CRL5807 cells, and its IC50 value is found to be
0.26 0.2 µg/ml. Compounds 3e and 3f have also affected MDA‐
MB231 cells more than other cells, and their IC50 values were found
to be 12.81 1.4 and 2.00 0.4 µg/ml, respectively.
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2
RESULTS AND DISCUSSION
Chemistry
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2.1
The synthesis of coumarin derivatives containing the triazole ring
(3a–f) was performed according to the pathways outlined in
Scheme 1. The starting compounds 1a,b and 2a–c were synthesized
according to a previously reported study.[27] Coumarin‐3‐carboxylic
acid and 6‐chloro‐coumarin‐3‐carboxylic acid were prepared by the
reaction of the respective salicylaldehydes and 2,2‐dimethyl‐1,
3‐dioxane‐4,6‐dione in ethanol containing pyridine, which was trea-
ted with SOCl2 to give starting materials 1a,b. Treatment of com-
pounds hydrazones with hydrazine monohydrate in water resulted in
the synthesis of 4‐amino‐1,2,4‐triazol‐3‐one derivatives 2a–c, which
is the second intermediate. Finally, coumarin–triazole derivatives
were obtained from the reaction of compounds 1a,b with molecules
2a–c under reflux in benzene. Spectral investigations of synthesized
compounds were carried out in accordance with the proposed
structures of target molecules.
The 1H nuclear magnetic resonance (NMR) spectra of synthe-
sized compounds showed characteristic peaks for NH (triazole N‐2)
and CONH at 11.67–11.78 and 10.90–10.97 ppm, respectively.
Coumarin H‐4 signals were observed between 8.38 and 8.83 ppm. In
the 13C NMR spectra of compounds 3a–f, triazole C‐3 and coumarin
C‐5 signals were observed between 153.13 and 154.68 and 146.52
and 147.23 ppm, respectively. Coumarin C‐2 and C‐4 signals were
shown at about 159.13–159.59 and 148.37–149.56 ppm, respec-
tively. In the mass spectral data, suitable molecular ion peaks were
observed according to the molecular formulae of compounds 3a–f.
It can be remarked that these compounds show higher inhibition
effects on PANC‐1 cells as compared with doxorubicin. Moreover,
their overall high inhibition ability against MDA‐MB231 cells leads to
more detailed research on breast cancer.
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2.3
Molecular docking study
The docking study revealed that the docking scores of coumarin
derivatives 3a–f are in the range of −8.285 to −9.673 kcal/mol as
compared with the docking score of doxorubicin, −15.354 kcal/mol,
at the same catalytic site of human topoisomerase IIα. These results
indicate that most of the studied compounds have a similar binding
pattern with binding site residues of doxorubicin. The significant
binding affinity toward human topoisomerase IIα implies that these
compounds are very active against cancer cells.
The detailed analysis of the docking study reveals that hydrogen
bonds and hydrophobic interactions predominate in protein–ligand
interactions, which are shown in Table 2 and Figure 2 with a two‐
dimensional diagram. The docking simulation suggests that com-
pound 3b was buried in the active site of the enzyme, forming hy-
drogen bond interactions with Asn91, Asp94, Ser149, and Asn150
TABLE 1 IC50 values of cell lines for different formulations (µg/ml)
CCD34Lu
48.48 2.6
5.47 0.5
28.96 7.6
2.43 0.9
<50
CRL5807
CRL5826
MDA‐MB231
<50
HTB177
<50
PC‐3
PANC‐1
3a
39.69 4.2
1.37 0.01
15.17 6.1
0.26 0.2
21.13 3.1
2.23 0.6
10.76 0.5
44.11 5.63
1.59 0.5
9.4435 1.9
0.97 0.1
27.4 11.7
2.41 0.8
6.85 0.3
–
43.80 11.8
4.56 1.2
15.86 0.02
10.25 3.8
44.93 1.8
7.57 0.7
<20
3b
0.86 0.04
4.10 0.4
0.60 0.3
12.81 1.4
2.00 0.4
16.01 0.8
4.48 1.5
<50
6.04 2.7
<50
3c
3d
8.32 3.1
<50
10.04 5.4
<50
3e
3f
36.61 21.4
18.87 0.9
9.20 3.0
2.90 0.2
11.40 5.5
17.6 0.9
Doxorubicin

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TABLE 2 The docking scores and interacting residues of studied compounds
Docking score
Title
3a
(kcal/mol)
H‐bond residue (distance, Å)
Hydrophobic interactions residue (distance, Å)
−8.285
Asn91 (2.01), Asp94 (2.40), Ser149 (3.45),
Asn150 (1.91)
Asn95 (3.86), Ile141 (3.67), Ile141 (3.41), Phe142 (3.81),
Thr215 (3.79)
3b
3c
3d
−8.882
−9.673
−8.622
Asn91 (1.98), Asp94 (2.14), Ser149 (2.35),
Asn150 (2.08), Ser149 (3.54)
Asn91 (3.83), Asn95 (3.89), Ile141 (3.98), Ile141 (3.66),
Ile141 (3.50), Phe142 (3.98), Thr215 (3.79)
Asn91 (1.78), Asp94 (2.32), Ser149 (2.12),
Asn150 (2.14), Ser149 (3.55), Ser149 (3.57)
Asn91 (3.64), Ile141 (3.40), Thr215 (3.71)
Asn91 (2.89), Asp94 (2.17), Ser149 (3.60),
Asn150 (2.04)
Ile141 (3.61), Ile141 (3.65), Ile141 (3.27), Thr215 (3.93)
3e
3f
−8.837
−9.212
Asn91 (2.08), Asp94 (2.19), Asn150 (1.81)
Ile141 (3.28), Phe142 (3.94), Thr215 (3.74), Ile217 (3.85)
Asn91 (3.94), Ile141 (3.29), Ile141 (3.68), Thr215 (3.56)
Asn91 (1.79), Asp94 (2.22), Ser149 (2.40),
Asn150 (2.00)
Doxorubicin −10.666
Asn91 (2.06), Asn95 (1.94), Arg98 (1.94), Asn120 Thr159 (3.24), Asn91 (3.73), Ala92 (3.89), Arg98 (3.91),
(2.65), Asn150 (1.80)
Ile125 (3.59), Phe142 (3.94), Ile217 (3.61)
residues. The same interactions were also observed with the other
compounds and doxorubicin.
Mercury spectrometer. The mass spectra were recorded on Agilent
1260 Infinity series Accurate‐Mass Time‐of‐Flight (TOF) LC/MS
spectrometer.
The InChI codes of the investigated compounds, together
with some biological activity data, are provided as Supporting
Information.
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3
CONCLUSION
In this study, a new series of novel coumarin derivatives containing a
triazole ring was designed and synthesized. The newly synthesized
molecules were screened for their in vitro anticancer activity against
CRL5807, CRL5826, MDA‐MB231, HTB177, PC‐3, PANC‐1 cell lines,
used as cancer cells, and CCD34Lu, as a healthy cell line. Among the
tested molecules, compounds 3b and 3d showed significant cyto-
toxicity against all the cancer cell lines and their most affected cells,
respectively, MDA‐MB231 and CRL5807 cells. Compounds 3b and
3d showed higher antitumor inhibitory activities against the
CCD34Lu cell line than the standard doxorubicin drug. Molecules
3b–f exhibited a significant antitumor inhibitory activity against the
MDA‐MB231 cell lines as compared with the reference drug dox-
orubicin. The docking study suggests that the compounds can be
considered as potential leads against cancer, with significant inter-
actions with human topoisomerase IIα. This study could be utilized
for the design of an effective drug for the treatment of cancer.
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4.1.2
Synthesis of compounds 3a–f
A solution of compounds 2a–c (0.01 mol) in dry benzene (10 ml) and
compounds 1a,b (0.01 mol) was taken in a round‐bottom flask. The
mixture was refluxed for 2 hr. After the completion of the reaction
(TLC, eluent ethylacetate/hexane, 3:1), the mixture was cooled to
room temperature; the product appeared as a white solid. It was
filtrated and washed with ethanol to obtain the pure product.
N‐[3‐(4‐Fluorobenzyl)‐5‐oxo‐1,5‐dihydro‐4H‐1,2,4‐triazol‐4‐yl]‐2‐
oxo‐2H‐chromene‐3‐carboxamide (3a)
Yield: 85%, m.p.: 311–312°C, 1H NMR (dimethyl sulfoxide [DMSO]‐
d6), δ, ppm: 3.79 (2H, s, CH2); 7.08 (2H, t, J = 8.0 Hz, Ar‐H); 7.27–7.30
(2H, m, Ar‐H); 7.47 (1H, t, J = 4.0 Hz, Ar‐H); 7.54 (1H, d, J = 8.0 Hz, Ar‐
H); 7.81 (1H, t, J = 8 Hz, Ar‐H); 8.00 (1H, d, J = 8 Hz, Ar‐H); 8.83 (1H,
s, CH, coumarin‐H4); 10.90 (1H, s, CONH); 11.67 (1H, s, NH). 13C
attached proton test (APT; DMSO‐d6), δ, ppm: 30.39 (CH2); 115.49
(d, JCF = 22 Hz); 116.76; 118.04; 118.56; 125.77; 131.02; 131.11;
131.37; 131.45; 135.31 (Ar‐C); 147.23 (C═N); 149.56 (CH, coumarin‐
C4); 152.85 (coumarin‐C3); 154.68 (C═O, triazole); 159.94 (C═O,
coumarin C2); 161.72 (C═O); 164.64 (C‐F, d, J = 241 Hz). LC–MS, m/
z: 381.1018 [M+H]+.
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4
EXPERIMENTAL
Chemistry
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4.1
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4.1.1
General
The reaction progress was monitored by thin‐layer chromatography
(TLC) on silica gel plates (Merck 60, F254, 0.2 mm). The melting points
were determined on capillary tubes using the Stuart SMP30 melting
point apparatus and were uncorrected. 1H and 13C NMR spectra
(400 and 100 MHz, respectively) were obtained using a Varian
6‐Chloro‐N‐[3‐(4‐fluorobenzyl)‐5‐oxo‐1,5‐dihydro‐4H‐1,2,4‐triazol‐
4‐yl]‐2‐oxo‐2H‐chromene‐3‐carboxamide (3b)
Yield: 83%, m.p.: 347°C (decomp.), 1H NMR (DMSO‐d6), δ, ppm: 3.78
(2H, s, CH2); 7.09 (2H, t, J = 4.0 Hz, Ar‐H); 7.27 (2H, t, J = 8 Hz, Ar‐H);

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FIGURE 2 Two‐dimensional interaction diagrams of compounds 3b, 3c, and doxorubicin at the catalytic site of human topoisomerase IIα
7.59 (1H, t, J = 8.0 Hz, Ar‐H); 7.84 (1H, d, J = 8.0 Hz, Ar‐H); 8.16 (1H, s,
Ar‐H); 8.81 (1H, s, CH, coumarin‐H4); 10.97 (1H, s, CONH); 11.74 (1H,
s, NH). 13C APT (DMSO‐d6), δ, ppm: 30.31 (CH2); 115.53 (d, JCF = 21
Hz); 118.83; 119.21; 119.95; 129.41; 129.83; 131.35; 131.41; 131.49;
134.68 (Ar‐C); 147.21 (C═N); 148.37 (CH, coumarin‐C4); 152.77
(coumarin‐C3); 153.31 (C═O, triazole); 159.13 (C═O, coumarin C2);
161.42 (C═O); 161.62 (C‐F, d, J = 241 Hz). LC–MS, m/z: 415.0627 [M
(Cl35)+H]+, 417.0598 [M (Cl37)+H]+.
116.76; 117.73; 118.57; 125.80; 128.79; 129.97; 130.06; 130.90;
131.10; 131.25; 131.61; 135.42; 136.41 (Ar‐C); 146.57 (C═N);
149.80 (CH, coumarin‐C4); 152.79 (coumarin‐C3); 154.68 (C═O,
triazole); 159.59 (C═O, coumarin C2); 161.58 (C═O). LC–MS, m/z:
415.0627 [M (Cl35)+H]+, 417.0598 [M (Cl37)+H]+.
6‐Chloro‐N‐[3‐(3,4‐dichlorobenzyl)‐5‐oxo‐1,5‐dihydro‐4H‐1,2,4‐
triazol‐4‐yl]‐2‐oxo‐2H‐chromene‐3‐carboxamide (3d)
Yield: 85%, m.p.: >320°C, 1H NMR (DMSO‐d6), δ, ppm: 3.83 (2H, s,
CH2); 7.24 (1H, d, J = 8 Hz, Ar‐H); 7.50 (1H, d, J = 8 Hz, Ar‐H); 7.53
(1H, s, Ar‐H); 7.58 (1H, d, J = 8 (Hz, Ar‐H); 7.82 (1H, d, J = 8 Hz, Ar‐H);
8.14 (1H, s, Ar‐H); 8.79 (1H, s, CH, coumarin‐H4); 10.94 (1H, s,
CONH); 11.74 (1H, s, NH). 13C APT (DMSO‐d6), δ, ppm: 30.20 (CH2);
118.81; 119.01; 119.93; 129.45; 129.85; 130.00; 130.03; 130.90;
131.28; 131.58; 134.73; 136.40 (Ar‐C); 146.52 (C═N); 148.49 (CH,
N‐[3‐(3,4‐Dichlorobenzyl)‐5‐oxo‐1,5‐dihydro‐4H‐1,2,4‐triazol‐4‐yl]‐
2‐oxo‐2H‐chromene‐3‐carboxamide (3c)
Yield: 86%, m.p.: 311–312°C, 1H NMR (DMSO‐d6), δ, ppm: 3.83 (2H,
s, CH2); 7.46–7.56 (4H, m, Ar‐H); 7.81 (2H, t, J = 8 Hz, Ar‐H); 8.01
(1H, d, J = 8.0 Hz, Ar‐H); 8.82 (1H, s, CH, coumarin‐H4); 10.94 (1H, s,
CONH); 11.78 (1H, s, NH). 13C APT (DMSO‐d6), δ, ppm: 30.23 (CH2);

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coumarin‐C4); 152.75 (coumarin‐C3); 153.31 (C═O, triazole); 159.13
(C═O, coumarin C2); 161.35 (C═O). LC–MS, m/z: 464.9804 [M
(Cl35Cl35Cl35)+H]+, 466.9942 [M (Cl35Cl35Cl37)+H]+, 468.9910 [M
(Cl35Cl37Cl37)+H]+, 486.9796 [M (Cl35Cl35Cl35)+Na].
4.2.2
Cytotoxicity assay
All cell lines were inoculated in a 96‐well plate at 1 × 105 cells/ml
initial cell concentration. Cells were treated with increasing
concentrations of samples (0.5, 5, and 50 µg/ml) for 48 hr, and
doxorubicin was used as the positive control (20, 2, and
0.2 µg/ml; see Figure S2). Cell viabilities of increasing con-
centrations of doxorubicin for different cells (0.2, 2, 20 µg/ml)
were determined. At the end of the 48 hr, 20 μl MTT (from
2.5‐mg/ml stock) was added to each well and cells were
incubated for a further 4 hr at 37°C in a 95% humidified atmo-
sphere of 5% CO2. The medium was replaced with 150 μl DMSO
for dissolving formazan crystals. Optical densities of the
dissolved material were determined at 570 nm (reference filter,
620 nm) with UV–visible spectrophotometer. The percentage of
the viable cells was determined with the following formula:
N‐[3‐(3‐Chlorobenzyl)‐5‐oxo‐1,5‐dihydro‐4H‐1,2,4‐triazol‐4‐yl]‐2‐
oxo‐2H‐chromene‐3‐carboxamide (3e)
Yield: 87%, m.p.: 278–279°C, 1H NMR (DMSO‐d6), δ, ppm: 3.82 (2H,
s, CH2); 7.20–7.32 (4H, m, Ar‐H); 7,48 (1H, t, J = 8 Hz, Ar‐H); 7.55
(1H, d, J = 8.0 Hz, Ar‐H); 7.81 (1H, d, J = 8 Hz, Ar‐H); 8.02 (1H, d,
J = 8.0 Hz, Ar‐H); 8.38 (1H, s, CH, coumarin‐H4); 10.96 (1H, s,
CONH); 11.75 (1H, s, NH). 13C APT (DMSO‐d6), δ, ppm: 30.74 (CH2);
116.77; 117.85; 118.58; 125.79; 127.23; 128.34; 129.42; 130.65;
131.08; 133.33; 135.37; 137.73 (Ar‐C); 146.81 (C═N); 149.76 (CH,
coumarin‐C4); 152.79 (coumarin‐C3); 154.68 (C═O, triazole); 159.59
(C═O, coumarin C2); 161.59 (C═O). LC–MS, m/z: 397.0754 [M (Cl35
)
+H]+, 399.0728 [M (Cl37)+H]+.
% viable cells = (Asample/Acontrol) × 100,
6‐Chloro‐N‐[3‐(3‐chlorobenzyl)‐5‐oxo‐1,5‐dihydro‐4H‐1,2,4‐
triazol‐4‐yl]‐2‐oxo‐2H‐chromene‐3‐carboxamide (3f)
where Asample is the absorbance of the sample and Acontrol is the
absorbance of the control.
Yield: 86%, m.p.: 315–316°C, 1H NMR (DMSO‐d6), δ, ppm: 3.82
(2H, s, CH2); 7.20–7.30 (4H, m, Ar‐H); 7.59 (1H, d, J = 8 Hz, Ar‐H);
7.81 (1H, d, J = 8.0 Hz, Ar‐H); 8.15 (1H, s, Ar‐H); 8.78 (1H, s, CH,
coumarin‐H4); 10.96 (1H, s, CONH); 11.73 (1H, s, NH). 13C APT
(DMSO‐d6), δ, ppm: 30.71 (CH2); 118.82; 119.08; 119.94; 127.25;
128.33; 129.40; 129.43; 129.84; 130.66; 133.33; 134.71; 137.71
(Ar‐C); 146.76 (C═N); 148.47 (CH, coumarin‐C4); 152.75
(coumarin‐C3); 153.31 (C═O, triazole); 159.14 (C═O, coumarin
C2); 161.33 (C═O). LC–MS, m/z: 431.0354 [M (Cl35Cl35)+H]+,
433.0328 [M (Cl35Cl37)+H]+.
The half‐maximal inhibitory concentration (IC50) value is a con-
centration that inhibits cell viability by 50% under laboratory con-
ditions. The calculation of the IC50 was performed by using
GraphPad Prism 5 software.
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4.3
Protocol of the docking simulation
Protein coordinates of the crystal structure of human topoisomerase
IIα (PDB ID: 1ZXM) were obtained from the Protein Data Bank
(www.rcsb.org) with a resolution of 1.87 Å.[33] The docking simu-
lations were performed with the Maestro Molecular Modeling
platform (version 10.5) using Induced Fit Docking (IFD) protocol
combined with the Glide/XP method.[34] The protein preparation
protocol was performed to fill missing side chains, remove water
molecules, except within 5 Å in the active site, update missing
loop regions, and protonate residues at physiological pH.
The LigPrep tool was used to obtain the lowest energy
three‐dimensional structures of ligand molecules at neutral pH
and under the OPLS 2005 force field. All the conformations of
studied molecules were docked into the receptor grid of radii of
20 Å. The binding site of the receptor structure was centered on
the bonded ligand molecule.
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4.2
Cytotoxic activity assay
Cell culture conditions
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4.2.1
CRL5807 (human bronchioalveolar carcinoma), CRL5826 (human
squamous cell carcinoma), MDA‐MB231 (human breast cancer
cells), HTB177 (human lung cancer), PC‐3 (human prostate ade-
nocarcinoma), PANC‐1 (human pancreatic cancer cells) cell lines,
used as cancer cells, and CCD34Lu (normal human lung fibro-
blasts; American Type Culture Collection), used as a healthy cell
line. All cell lines were cultured in Dulbecco's modified Eagle's
medium: nutrient mixture F‐12 with 10% fetal bovine serum and
0.1% penicillin/streptomycin (Serox GmbH). Cells were incubated
at 37°C in a 95% humidified atmosphere of 5% CO2. Cytotoxicity
effects of samples were determined by the MTT (3‐(4,5‐dimethyl‐
2‐thiazolyl)‐2,5‐diphenyl‐2H‐tetrazolium bromide) assay, which
colorimetrically determines the activity of mitochondrial
reductase of viable cells.[1,32]
CONFLICTS OF INTERESTS
The authors declare that there are no conflicts of interests.
ORCID
Emre Menteşe
http://orcid.org/0000-0002-3408-8202

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REFERENCES
[20] M. J. Matos, D. Viña, C. Picciau, F. Orallo, L. Santana, E. Uriarte,
Bioorg. Med. Chem. Lett. 2009, 19, 5053.
[1] A. Güner, A. Nalbantsoy, A. Sukatar, N. Ü. Karabay Yavaşoğlu,
Cytotechnology 2019, 73, 687.
[21] D. Viña, M. J. Matos, M. Yáñez, L. Santana, E. Uriarte, Med. Chem.
Commun. 2012, 3, 213.
[2] A. Güner, E. Polatlı, T. Akkan, H. Bektaş, C. Albay, Turk. J. Chem.
2019, 43, 1270.
[22] S. K. Chauthe, S. Mahajan, M. Rachamalla, K. Tikoo, I. P. Singh, Med.
Chem. Res. 2015, 24, 2476.
[3] World Health Organization report, 2018. https://www.who.int/
health-topics/cancer#tab=tab_1. Accessed September 12, 2018.
[4] G. Feuer, Progress in Medicinal Chemistry, Vol. 10 (Eds: G. P. Ellis, G.
B. West), North Holland, New York, NY 1974, p. 86.
[5] F. M. Awadallah, T. A. El‐Waei, M. M. Hanna, S. E. Abbas, M. Ceruso,
B. E. Oz, O. O. Guler, C. T. Supuran, Eur. J. Med. Chem. 2015, 96, 425.
[6] H. M. T. Albuquerque, C. M. M. Santos, C. F. R. A. C. Lima,
L. M. N. B. F. Santos, J. A. S. Cavaleiro, A. M. S. Silva, Eur. J. Org.
Chem. 2017, 2017, 87.
[23] A. Thakur, R. Singla, V. Jaitak, Eur. J. Med. Chem. 2015, 101, 476.
[24] Y. L. Fan, X. Ke, M. Liu, J. Heterocycl. Chem. 2018, 55, 791.
[25] S. A. Morsy, A. A. Farahat, M. N. A. Nasr, A. S. Tantawy, Saudi Pharm.
J. 2017, 25, 873.
[26] C. F. Liu, Q. K. Shen, J. J. Li, Y. S. Thian, Z. Quan, J. Enzyme Inhhib.
Med. Chem. 2017, 32, 1111.
[27] B. Kahveci, F. Yılmaz, E. Menteşe, S. Ülker, Arch. Pharm. Chem. Life
Sci. 2017, 350, e1600369.
[28] R. Kaur, A. R. Dwivedi, B. Kumar, V. Kumar, Anti‐Cancer Agent Med.
Chem. 2016, 16, 465.
[7] M. E. Riveiro, A. Moglioni, R. Vazquez, N. Gomez, C. Davio, Bioorg.
Med. Chem. 2008, 16, 2665.
[29] D. Gupta, D. K. Jain, J. Adv. Pharm. Technol. 2015, 6, 141.
[30] R. Paprocka, M. Wiese, A. Eljaszewicz, A. Helmin‐Basa, A. Gzella,
B. Modzelewska‐Banachiewicz, J. Michalkiewicz, Bioorg. Med. Chem.
Lett. 2015, 25, 2664.
[8] O. Prakash, R. Kumar, V. Parkash, Eur. J. Med. Chem. 2008, 43, 435.
[9] R. R. Zhang, J. Liu, Y. Zhang, M. Q. Hou, M. Z. Zhang, F. Zhou,
W. H. Zhang, Eur. J. Med. Chem. 2016, 116, 76.
[10] M. Z. Zhang, R. R. Zhang, J. Q. Wang, X. Yu, Y. L. Zhang, Q. Q. Wang,
W. H. Zhang, Eur. J. Med. Chem. 2016, 124, 10.
[31] S. Maddila, R. Pagadala, S. B. Jonnalagadda, Lett. Org. Chem. 2013,
10, 693.
[11] P. Dai, X. Yu, P. Teng, W. H. Zhang, C. Deng, Org. Lett. 2018,
20, 6901.
[32] T. Mosmann, J. Immunol. Methods 1983, 65, 55.
[33] H. Wei, A. Ruthenburg, S. Bechis, G. Verdine, J. Biol. Chem. 2005,
280(44), 37041.
[12] A. Gomes, E. Fernandes, A. M. S. Silva, D. C. G. A. Pinto,
C. M. M. Santos, J. A. S. Cavaleiro, J. L. F. C. Lima, Biochem.
Pharmacol. 2009, 78, 171.
[34] Schrödinger Release, 1: Maestro, version 10.1, Schrödinger, LLC,
New York, NY, 2015.
[13] M. T. Flavin, J. D. Rizzo, A. Khilevich, A. Kucherenko, A. K. Sheinkman,
V. Vilaychack, L. Lin, W. Chen, E. M. Greenwood, T. Pengsuparp,
J. M. Pezzuto, S. H. Hughes, T. M. Flavin, M. Cibulski, W. A. Boulanger,
R. L. Shone, Z. Q. Xu, J. Med. Chem. 1996, 39, 1303.
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[14] E. Menteşe, N. Baltaş, O. Bekircan, Arch. Pharm. Chem. Life Sci. 2019,
352, e1900227.
[15] X. T. Xu, X. Y. Deng, J. Chen, Q. M. Liang, K. Zhang, D. L. Li, P. P. Wu,
X. Zheng, R. P. Zhou, Z. Y. Jiang, A. J. Ma, W. H. Chen, S. H. Wang,
Eur. J. Med. Chem. 2020, 189, 112013.
How to cite this article: Menteşe E, Güner A, Polatlı E, Emirik
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2020;e2000284. https://doi.org/10.1002/ardp.202000284
[16] E. Menteşe, N. Karaali, G. Akyüz, F. Yılmaz, S. Ülker, B. Kahveci,
Chem. Heterocycl. Chem. 2016, 52(12), 1017.
[17] G. Bakhtiari, S. Moradi, S. Soltanali, Arabian J. Chem. 2014, 7, 972.
[18] X. Luo, J. Song, L. Cheng, D. Huang, Sci. China (Series B) Chem. 2001,
44(5), 532.
[19] M. P. Brun, L. Bischoff, C. Garbay, Angew. Chem., Int. Ed. 2004,
43, 3432.