Design, Synthesis, and Biological Evaluation of Coumarin–Triazole Hybrid Molecules as Potential Antitumor and Pancreatic Lipase Agents

Article abstract of DOI:10.1002/ardp.201600369

The design, synthesis, and investigation of antitumor and anti-lipase activities of some coumarin–triazole hybrid molecules are reported. The synthesis of these hybrid molecules was performed under microwave irradiation and conventional heating procedures

Full text of DOI:10.1002/ardp.201600369

ARC HH PHARM
Arch. Pharm. Chem. Life Sci. 2017, 350, e1600369
Archiv der Pharmazie
Full Paper
Design, Synthesis, and Biological Evaluation of Coumarin–Triazole
Hybrid Molecules as Potential Antitumor and Pancreatic Lipase
Agents
1
2
2
3
€
Bahittin Kahveci , Fatih Yılmaz , Emre Mente sS e
, and Serdar Ulker
1
Faculty of Health Sciences, Department of Nutrition and Diethetics, Karadeniz Technical University,
Trabzon, Turkey
Faculty of Arts and Sciences, Department of Chemistry, Recep Tayyip Erdogan University, Rize, Turkey
Faculty of Arts and Sciences, Department of Biology, Recep Tayyip Erdogan University, Rize, Turkey
2
3
The design, synthesis, and investigation of antitumor and anti-lipase activities of some coumarin–
triazole hybrid molecules are reported. The synthesis of these hybrid molecules was performed under
microwave irradiation and conventional heating procedures. The newly synthesized hybrid molecules
were investigated as inhibitors against four tumor cell lines (BT20 human breast carcinoma, SK-Mel
1
(
28 melanoma, DU-145 prostate carcinoma, and A549 lung carcinoma) and porcine pancreatic lipase
PPL). Most of these compounds showed notable antitumor activities against the tested tumor cell
lines, and compounds 8i and 8l showed the best anti-lipase activity of 99.30 ꢀ 0.56% and
9.85 ꢀ 1.21%, respectively, at a concentration of 10 mM.
9
Keywords: Antitumor / Coumarin / Hybrid molecule / Lipase inhibition / Triazol-3-one
Received: December 7, 2016; Revised: April 7, 2017; Accepted: April 17, 2017
DOI 10.1002/ardp.201600369
Additional supporting information may be found in the online version of this article at the publisher’s web-site.
:
Introduction
Overweight and obesity are globally the fifth leading risks for
death. Every year, about 2.8 million adults die from diseases
caused by obesity. Obesity and obesity-related diseases like
diabetes and coronary heart diseases are the major problem in
the world. An important way for obesity is treatment to
develop inhibitors of food digestion. Inhibition of pancreatic
lipase and the associated reduction of lipid absorption is an
attractive approach for the discovery of potent agents.
Orlistat is the only drug used as a pancreatic lipase inhibitor.
Unfortunately, it has some unpleasant side effects on human
body like oily stools, oily spotting and flatulence [4–6].
Coumarins (2H-chromen-2-one) are generally obtained
from plants and consists of a class of phenolic compounds.
Thirteen hundred coumarin derivatives have been identified
from plants, bacteria, and fungi. In the year of 1820, coumarin
was first obtained as a natural product and nearby 150 species
of coumarin were found in 30 diverse plant families such as
Umbelliferae, Oleaceae, Clusiaceae, Guttiferae, Rutaceae [7].
Coumarin and its derivatives show rare side effects on human
body [8]. Also, they are known to exhibit many pharmaco-
logical activities like antioxidant [9], anticoagulant [10],
Cancer comprises a group of diseases that include abnormal
cell growth over other parts of the human body. Cancer is a
big health problem and it is the second frequent cause of
death after heart diseases. It is characterized by uncontrolled
division and spread of abnormal cells in human body.
Although chemotherapy is widely used for various cancer
treatments, the toxicity and resistance of chemotherapeutic
drugs make it a necessity to design more selective drugs for
cancer therapy. For instance, the synthesis of effective novel
anticancer drugs is one of the most important targets of
modern medicinal chemistry [1–3].
Correspondence: Dr. Emre Mente sS e, Faculty of Arts and Sciences,
Department of Chemistry, Recep Tayyip Erdogan University, Rize
5
3100, Turkey.
E-mail: emre.mentese@erdogan.edu.tr
Fax: þ904642234019
ß 2017 Deutsche Pharmazeutische Gesellschaft
www.archpharm.com
(1 of 9) e1600369

Arch. Pharm. Chem. Life Sci. 2017, 350, e1600369
B. Kahveci et al.
ARC HH PHARM
Archiv der Pharmazie
antibacterial [11], antitubercular [12], antifungal [13], anti-
inflammatory [14], and anticancer [15]. In addition to these,
coumarin is regularly used as an important nucleus in
medicinal chemistry. Warfarin and acenocoumarol (Fig. 1)
are anticoagulant agents that are known as vitamin K
antagonists, and are normally used in the prevention of
thrombosis and thromboembolism [16].
Past researches have demonstrated that coumarin hydra-
zones have been investigated by researchers in recent years
due to the fact that they have pharmacologically powerful
properties [17, 18]. On the other hand, Nasr et al. [19]
reported in vitro anticancer activity of some coumarin
hydrazide-hydrazone derivatives and found that one of the
compounds could be a potent anticancer drug to overcome
drug resistance in cancer.
Triazole ring is an important five-membered heterocycle
structure for designing new bioactive molecules. This
electron-rich heterocycle easily binds to various types of
enzymes and receptors. Therefore, triazol containing
compounds show a broad spectrum of biological activi-
ties [20]. One of the isomers of this heterocycle is 1,2,4-
triazoles and they have been known to possess various
biological activities such as antimicrobial [21], anti-
inflammatory [22], and anticancer activities [23]. Some of
the examples of triazoles are in medical use, like ribavirin
Figure 2. Structures of some drugs bearing 1,2,4-triazole.
Results and discussion
Chemistry
0
The synthesis of target hybride molecules N -(1,2,4-triazol-1-
yl)acetyl coumarin-3-carbohydrazides (8) was performed by
reaction of 2-(3-alkyl/aryl-4-amino-5-oxo-4,5-dihydro-1H-
1,2,4-triazol-1-yl)acetohydrazides (5a–f) with 3-(1H-benzo-
triazol-1-ylcarbonyl)-2H-chromen-2-ones (7a–c). Firstly, com-
pounds 1a–f were prepared according to the related
literature [28]. Then, these compounds were reacted with
ethyl carbazate in ethanol to obtain corresponding hydra-
zone derivatives (2a–f). Next, compounds 2a–f were reacted
with hydrazine monohydrate in water to synthesize 4-amino-
1,2,4-triazol-3-one derivatives (3a–f). Ester derivatives of
compounds 3a–f were prepared by the reaction of com-
pounds 3a–f and ethyl bromoacetate in the presence of
sodium ethoxide (NaOEt) in ethanol. Afterwards, these
compounds were treated with hydrazine monohydrate in
n-butanol to obtain 2-(3-alkyl/aryl-4-amino-5-oxo-4,5-
dihydro-1H-1,2,4-triazol-1-yl)acetohydrazides (5a–f) [29],
which are the first intermediate for the synthesis of target
compounds.
(
antiviral) [24, 25], fluconazole (antifungal) [26], vorozole,
and letrozole (nonsteroidal competitive aromatase
inhibitor) [20] (Fig. 2).
Molecular modification is considered as an effective
method for developing new drug candidates [27]. Molecu-
lar hybridization is among the molecular modification
methods for achieving new molecules which have better
pharmacokinetic and pharmacodynamic properties. In
recent years, this method has become a powerful tool for
obtaining more bioactive compounds. Considering the
promising activity of coumarin and triazole moieties, the
use of these heterocycles in one molecule may result in the
formation of more bioactive compounds. Hence, we
decided to use the mentioned moieties in a molecular
hybridization approach to design new potential antitumor
and anti-lipase agents.
The synthesis of second intermediate 3-(1H-benzotriazol-1-
ylcarbonyl)-2H-chromen-2-ones (7a–c) was performed in
three steps. First, coumarin-3-carboxylic acid derivatives
(
6a–c) were prepared by the reaction of the respective
salicylic aldehydes and 2,2-dimethyl-1,3-dioxane-4,6-dione in
ethanol containing pyridine. Then, these compounds
were reacted with 1H-benzotriazole in the presence of SOCl
2
in dichloromethane to prepare 3-(1H-benzotriazol-1-
ylcarbonyl)-2H-chromen-2-ones (7a–c), which are the second
intermediate of the target compounds.
The synthesis of hybrid molecules resulted in the formation
of more bioactive compounds. This type of molecules can
offer the advantages of a combination therapy with improved
pharmacological properties [30, 31]. Our literature surveys
have revealed that benzotriazole is an easy leaving group and
Figure 1. Structures of warfarin and acenocoumarol.
ß 2017 Deutsche Pharmazeutische Gesellschaft
www.archpharm.com
(2 of 9) e1600369

Arch. Pharm. Chem. Life Sci. 2017, 350, e1600369
Coumarin–Triazole Hybrid Molecules
ARC HH PHARM
Archiv der Pharmazie
this group offers many advantages to synthesize new hybrid
molecules [29, 32]. Thus, we used this chemical transformation
method to synthesize the target compounds (8a–r) by the
reaction of compounds 5a–f and 7a–c (Scheme 1). This last
reaction step was performed under microwave irradiation
and using conventional heating procedure, and their results
were compared.
Compounds 8e, 8f, 8h, 8i, 8k, 8m, 8p, and 8r showed notable
anticancer activity against tested cell line. The compounds 8f,
8h, 8i, 8k, 8p, and 8r were found to be more potent against
human breast cancer cell line than cisplatin. Compound 8f
was found to be more potent against human prostate cancer
cell line than cisplatin. Also, compounds 8h and 8i were
found to be more potent against human lung cancer cell line
than cisplatin. The antitumor activity of the compounds
against human breast cancer cell line can be arranged in this
order compared to cisplatin 8k > 8i > 8h > 8p > 8f > 8r >
cisplatin > 8m > 8e. For prostate cell line this order was
determined as 8f > cisplatin > 8e. The antitumor activity of
the synthesized compounds against human lung cancer cell
line can be stated as 8h > 8i> cisplatin > 8r > 8m > 8e. Among
the tested compounds, compounds 8k and 8p inhibited
selectively (SI ¼ 5.2 and 2.7) human breast cancer cell line,
and compound 8m inhibited selectively human lung cancer
cell line (SI ¼ 2.6). All the tested compounds showed no
activity against melanoma.
The spectral results of the target compounds 8a–r corre-
1
spond to the proposed structures. In the H NMR spectra, two
NH signals (exchangeable with D
1.00 and 10.60 ppm and NH signal (exchangeable with D
was observed at about 5.30 ppm. The NCH signal was shown
2
O) were observed at about
1
2
2
O)
2
13
at about 4.45 ppm. In the C NMR spectra, C–O signals were
shown at about 165.0, 160.0, 159.0, and 153.0 ppm as two
hydrazides, coumarin C-2, triazole C-3, and the C–N signal was
observed at about 147.0 ppm. In addition, all compounds
showed the proposed molecular ion signals in their LC-MS
spectra.
Biological activity
Cytotoxicity
Lipase inhibition assay
The compounds 8a–r were screened for in vitro anticancer
activity against normal human fibroblasts and four human
tumor cell lines: BT20 human breast carcinoma, SK-Mel 128
melanoma, DU-145 prostate carcinoma, and A549 lung
carcinoma, in 24-h drug exposure assays using cisplatin as
reference. The synthesized compounds’ cytotoxicity proper-
ties were determined using tetrazolium dye reduction
assay [33–35]. The cytotoxic activities of compounds are
displayed in Table 1. As shown in Table 1, the tested
compounds showed moderate to good activity in the in vitro
antitumor screening expressed by the CC50 values.
All compounds were evaluated with regard to pancreatic
lipase activity and most of compounds showed anti-lipase
activities at various concentrations (Table 2). Among the
tested compounds 8i and 8l showed the best anti-lipase
activity. These compounds inhibited pancreatic lipase by
99.30 ꢀ 0.56% and 99.85 ꢀ 1.21% at concentration of 10 mM,
respectively (Table 2). Orlistat, known as a pancreatic lipase
inhibitor used as an anti-obesity drug, showed inhibitory
effect by 99.88 ꢀ 0.43% at concentration of 300 nM
(IC50 ¼ 0.41 ꢀ 0.01 nM). Compound 8i and 8l IC50 values were
calculated as 2.64 ꢀ 0.33 mM and 1.80 ꢀ 0.08 mM, respectively.
Scheme 1. Synthetic route of compounds 8a–r.
ß 2017 Deutsche Pharmazeutische Gesellschaft
www.archpharm.com
(3 of 9) e1600369

Arch. Pharm. Chem. Life Sci. 2017, 350, e1600369
B. Kahveci et al.
ARC HH PHARM
Archiv der Pharmazie
Table 1. The antitumor activity of synthesized compounds against different tumor and normal cell lines.
CC50 (Cytotoxicity 50: mg/mL); SI (Selectivity index)
Compd.
BT-20
SI
DU-145
SI
SK-MEL128
SI
A549
SI
HFC
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
8
a
b
c
d
e
f
g
h
i
j
k
l
m
n
o
p
q
r
32.6
11.6
15.6
10.2
27.6
13.9
23.1
10.0
7.3
31.7
6.4
12.8
11.1
19.0
10.6
3.6
0.2
1.0
0.9
0.8
1.2
2.6
1.1
3.1
3.6
1.1
5.2
0.7
1.4
0.9
0.3
2.7
1.3
2.4
2.0
30.1
16.8
3.9
14.8
4.1
0.2
0.7
3.5
0.5
8.2
9.9
0.7
1.8
4.4
1.4
3.0
0.7
4.9
1.0
0.5
1.0
1.9
3.9
8.4
6.8
10.5
18.4
8.6
18.7
12.3
12.5
32.7
20.6
29.4
36.7
12.1
7.5
18.9
4.1
6.1
10.1
18.8
0.7
1.0
1.1
0.7
0.9
1.8
3.0
2.1
1.0
1.3
1.2
0.9
0.8
2.1
0.9
0.8
1.6
0.9
1.4
15.4
15.9
14.7
25.1
6.4
13.8
28.1
23.0
7.5
0.4
0.8
0.5
1.2
2.4
1.3
1.1
4.2
3.3
0.9
2.6
0.8
2.6
1.1
0.6
1.5
0.8
2.9
3.0
6.5
12.0
13.7
7.7
33.3
36.6
26.0
31.1
26.3
35.0
33.3
9.3
15.4
17.8
3.3
9.9
8.9
3.7
36.0
17.5
6.1
24.3
10.9
12.9
3.1
17.4
6.4
10.1
4.6
6.7
8.1
37.9
12.7
11.3
6.0
16.7
5.2
6.8
11.6
9.2
3.8
6.9
10.9
5.8
26.2
11.5
Cisplatin
1.4
Synthesized compounds 8i and 8l have a potential to be
alternatives to orlistat.
yields and time for us. These compounds were screened for
antitumor and anti-lipase activities. The anti-lipase activity
results indicate that compounds 8i and 8l showed the best
anti-lipase activity by 99.30 ꢀ 0.56% and 99.85 ꢀ 1.21% at
concentration of 10 mM, respectively. Antitumor activity of
the synthesized compounds was investigated against four
tumor cell lines (BT20 human breast carcinoma, SK-Mel 128
melanoma, DU-145 prostate carcinoma, and A549 lung
carcinoma). The results of this activity show that compounds
8f, 8h, 8i, 8k, 8q, and 8r were found to be more active than
standard cisplatin against BT20 human breast carcinoma
tumor cells, compound 8f was found to be more active than
standard cisplatin against DU-145 prostate carcinoma and
compounds 8h and 8i were found to be more active than
standard cisplatin against A549 lung carcinoma. Most of the
hybrid molecules showed more selectivity index (SI) value
than cisplatin. It means that the hybrid molecules have more
selectivity to show effects on cancer cells than normal human
cells. Based on the biological activity, these results indicate
that coumarin–triazol hybrid molecules show more activity
than coumarin (6a–c, 7a–c) and triazol (3a–f, 4a–f, 5a–f)
derivatives. These results have shown that combination of
triazol (5a–f) and coumarin (7a–c) has a positive effect on
biological activity of this type of molecules.
Conclusion
In this work, a new series of novel coumarin–triazol hybrid
molecules was designed and synthesized by using microwave
irradiation and conventional heating techniques. It was seen
that the synthesis of these compounds by microwave
irradiation method has supplied advantages on reaction
Table 2. Inhibitory effects of selected synthesized
compounds (synthesized compounds at final concentra-
tion of 10 mm).
Compounds
% Inhibition
IC50 (mM)
8
8
8
8
8
8
8
8
8
8
8
b
c
d
f
h
i
j
k
l
80.74 ꢀ 5.60
96.19 ꢀ 1.37
81.96 ꢀ 3.96
95.07 ꢀ 2.73
99.38 ꢀ 0.18
97.81 ꢀ 2.03
99.30 ꢀ 0.56
99.04 ꢀ 2.29
99.85 ꢀ 1.21
82.93 ꢀ 6.45
90.63 ꢀ 1.27
99.88 ꢀ 0.43
5.78 ꢀ 3.08
3.94 ꢀ 1.43
3.47 ꢀ 0.22
3.29 ꢀ 0.03
4.11 ꢀ 1.10
3.45 ꢀ 0.21
2.64 ꢀ 0.33
6.91 ꢀ 0.51
1.80 ꢀ 0.08
4.96 ꢀ 1.81
4.36 ꢀ 0.10
0.41 ꢀ 0.01 (nM)
Experimental
n
p
Orlistat
Chemistry
General
Orlistat (at final concentration of 0.3 mm) was used as positive
All reaction progress was monitored by TLC on silica gel plates
control.
(Merck 60, F254, 0.2 mm). The melting points were determined
ß 2017 Deutsche Pharmazeutische Gesellschaft
www.archpharm.com
(4 of 9) e1600369

Arch. Pharm. Chem. Life Sci. 2017, 350, e1600369
Coumarin–Triazole Hybrid Molecules
ARC HH PHARM
Archiv der Pharmazie
on capillary tubes on Stuart SMP30 melting point apparatus
mixture was cooled to room temperature and the product
appeared as a white solid. It was filtrated and washed with
ethanol to obtain the pure product.
1
13
and uncorrected. H and C NMR spectra (400 and 100 MHz,
respectively) were obtained using a Varian-Mercury (tetra-
methylsilane as internal standard): chemical shifts are
expressed in d values (ppm). The mass spectra were recorded
on Agilent 1260 Infinity series accurate-mass time-of-flight
Microwave method: Compounds 5a–f (0.01 mol) and com-
pounds 7a–c (0.011 mol) were taken in a microwave process
vial and dry ethanol (5 mL) was added. Then, mixture was
irradiated in microwave at 135°C for 30 min at 300 W
maximum power. After the completion of the reaction, the
mixture was taken in the beaker with hot ethanol, and a
product appeared as a white solid. It was filtrated and washed
with ethanol to obtain the pure product.
(
TOF) LC/MS spectrometer. All experiments were carried out in
microwave process vials (30 mL) with control of the tempera-
ture by infrared detection temperature sensor. It was
monitored by a computer and maintained constant at a
constant value by
a discrete modulation of delivered
microwave power. After completion of the reaction, the vial
was cooled to 60 °C via air jet cooling. The compounds 5a–f
and 7a–c were prepared according to the literature
procedure [29].
The H NMR, C APT, and LC-MS spectra and the InChI
codes of the investigated compounds together with some
biological activity data are provided as Supporting
Information.
0
N -[(4-Amino-3-methyl-5-oxo-4,5-dihydro-1H-1,2,4-
triazol-1-yl)acetyl]-6-chloro-2-oxo-2H-chromen-3-
carbohydrazide (8a)
Yield: 2.37 g (60% for microwave), 1.50 g (38% for
conventional), m.p.: 265–266°C, H NMR (DMSO-d
1
13
1
6
), d, ppm:
11.04 (1H, s, NH); 10.60 (1H, s, NH); 8.78 (1H, s, H-4); 8.08 (1H, d,
J ¼ 2.4 Hz, Ar-H); 7.74 (1H, dd, J ¼ 8.8 Hz, J ¼ 2.4 Hz, Ar-H); 7.26
(
1H, d, J ¼ 8.8 Hz, Ar-H); 5.25 (2H, s, NH
2 2
); 4.42 (2H, s, NCH );
1
3
Synthesis of compounds 7a–c
2 6
2.10 (2H, s, CH ). C APT (DMSO-d
), d, ppm: 164.7 (C–O);
These compounds were synthesized according to the litera-
ture procedure [29].
159.8 (C–O); 158.7 (CO coumarin); 153.7 (coumarin C-3); 152.9
(C–O triazole); 147.1 (coumarin C-4); 145.6 (C–N); 134.2;
1
29.5; 120.2; 119.6; 118.7 (Ar-C); 46.4 (NCH
2
); 11.1 (CH
3
). LC-
3
5
þ
37
þ
3
-(1H-Benzotriazol-1-ylcarbonyl)-6-chloro-2H-chromen-2-
on (7a)
Yield: 2.20 g (63%), m.p.: 248–249°C, H NMR (DMSO-d
MS, m/z: 393.0325 [M (Cl )þH] , 395.0692 [M (Cl )þH] ,
3
5
þ
37
þ
415.0596 [M (Cl )þNa] , 417.0529 [M (Cl )þNa] .
1
6
), d,
0
ppm: 8.74 (1H, s, H-4); 8.29–8.05 (2H, m, Ar-H); 8.05 (1H, d,
J ¼ 2.4 Hz, Ar-H); 7.98–7.79 (2H, m, Ar-H); 7.68–7.64 (1H, m, Ar-
N -{[4-Amino-3-(4-chlorobenzyl)-5-oxo-4,5-dihydro-1H-
1,2,4-triazol-1-yl]acetyl}-6-chloro-2-oxo-2H-chromen-3-
carbohydrazide (8b)
Yield: 3.18 g (63% for microwave), 2.27 g (45% for
conventional), m.p.: 259–260°C, H NMR (DMSO-d
13
H); 7.64 (1H, d, J ¼ 8.0 Hz, Ar-H). C APT (DMSO-d
6
), d, ppm:
1
1
1
62.8 (C–O); 157.3 (C–O coumarin C-2); 153.2 (coumarin C-3);
46.9 (coumarin C-4); 145.9; 134.4; 131.7; 130.9; 129.5; 127.5;
1
6
), d, ppm:
22.6; 120.7; 119.3; 119.1; 114.2 (Ar-C). LC-MS, m/z: 348.0203
11.02 (1H, s, NH); 10.62 (1H, s, NH); 8.81 (1H, s, H-4); 8.11 (1H, d,
35
37
[
M (Cl )þNa], 350.0156 [M (Cl )þNa].
J ¼ 2.4 Hz, Ar-H); 7.77 (1H, dd, J ¼ 9.2 Hz, J ¼ 2.4 Hz, Ar-H); 7.54
(
1H, d, J ¼ 9.2 Hz, Ar-H); 7.35 (2H,m, Ar-H); 7.29 (2H, m, Ar-H);
13
3
-(1H-Benzotriazol-1-ylcarbonyl)-6,8-dichloro-2H-
2 2 2
5.30 (2H, s, NH ); 4.45 (2H, s, NCH ); 3.88 (2H, s, CH ). C APT
chromen-2-on (7b)
Yield: 2.34 g (65%), m.p.: 263–264°C, H NMR (DMSO-d
ppm: 8.76 þ 8.67 (1H, s, coumarin C-4); 8.29 (1H, s, coumarin H-
), 8.27 (1H, s, coumarin H-5), 8.11–7.99 (2H, m, Ar-H), 7.85
(DMSO-d
6
), d, ppm: 164.6 (C–O); 159.8 (CO); 158.8 (C–O);
153.8 (coumarin C-3); 152.9 (CO triazole); 147.2 (CN); 147.0
(coumarin C-4); 135.2; 134.2; 313.8; 131.1; 129.6; 129.3; 128.7;
1
6
), d,
7
2 2
120.1; 119.7; 118.7 (Ar-C); 46.5 (NCH ); 30.2 (CH ). LC-MS, m/z:
1
3
35
þ
37
þ
(
(
1
1
1H, t, J ¼ 7.2 Hz, Ar-H), 7.67 (1H, t, J ¼ 7.2 Hz, Ar-H). C APT
DMSO-d
), d, ppm: 163.9 (C–O); 158.1 (C–O coumarin C-2);
52.4 (coumarin C-3); 148.7 (coumarin C-4); 146.8; 138.4;
33.1; 132.4; 129.9; 128.4; 123.1; 121.9; 116.0; 1114.1; 112.0
503.0605 [M (Cl )þH] , 505.0577 [M (Cl )þH] , 525.0418 [M
35 þ 37 þ
6
(Cl )þNa] , 527.0392 [M (Cl )þNa] .
0
N -{[4-Amino-3-(4-bromobenzyl)-5-oxo-4,5-dihydro-1H-
35
35
35
(
(
Ar-C). LC-MS, m/z: 359.0207 [M (Cl )(Cl )], 361.0153 [M (Cl
)
1,2,4-triazol-1-yl]acetyl}-6-chloro-2-oxo-2H-chromen-3-
carbohydrazide (8c)
Yield: 3.57 g (65% for microwave), 2.47 g (45% for
conventional), m.p.: 281–282°C, H NMR (DMSO-d
37
Cl )].
-(1H-Benzotriazol-1-ylcarbonyl)-7-diethylamino-2H-
1
3
6
), d, ppm:
chromen-2-on (7c)
11.02 (1H, s, NH); 10.62 (1H, s, NH); 8.81 (1H, s, H-4); 8.10 (1H, d,
Yield: 2.46 g (68%), m.p.: 210–211°C (lit. 212–214°C [36]).
J¼ 2.4 Hz, Ar-H); 7.77 (1H, dd, J ¼ 9.2 Hz, J¼ 2.4 Hz, Ar-H); 7.53
(
1H, d, J ¼ 9.2 Hz, Ar-H); 7.35 (2H,d, J¼ 8.4 Hz, Ar-H); 7.23 (2H, d,
Synthesis of compounds 8a–r
J¼ 8.4 Hz, Ar-H); 5.30 (2H, s, NH
2
); 4.45 (2H, s, NCH
), d, ppm: 164.6 (C –– O); 159.8 (C –– O); 158.7
2
); 3.68 (2H, s,
1
3
Conventional method:
(
(
A
solution of compounds 5a–f
0.01 mol) in ethanol (15 mL) and compounds 7a–c
0.011 mol) was taken in a round-bottom flask. The mixture
CH ). C APT (DMSO-d
2 6
(C –– Ocoumarin);153.8(coumarinC-3);152.9(C –– Otriazole);147.2
(CN); 147.0 (coumarin C-4); 135.7; 134.2; 131.6; 131.5; 129.6;
was refluxed for 8 h. After the completion of the reaction, the
2 2
129.3;120.3;120.0;119.7;118.7(Ar-C);46.5(NCH );30.2(CH ). LC-
ß 2017 Deutsche Pharmazeutische Gesellschaft
www.archpharm.com
(5 of 9) e1600369

Arch. Pharm. Chem. Life Sci. 2017, 350, e1600369
B. Kahveci et al.
ARC HH PHARM
Archiv der Pharmazie
3
5
79
þ
35
81
MS, m/z: 547.0603 [M (Cl )(Br )þH] , 549.0147 [M (Cl )(Br )þ
10.98 (1H, s, NH); 10.55 (1H, s, NH); 8.79 (1H, s, H-4); 8.11 (1H, d,
þ
37
81
þ
H] , 551.3242 [M (Cl )(Br )þH] .
J ¼ 2.4 Hz, Ar-H); 8.07 (1H, d, J ¼2.4 Hz, Ar-H); 5.25 (2H, s, NH
2
);
1
3
4
.40 (2H, s, NCH ); 2.07 (3H, s, CH ). C APT (DMSO-d ), d, ppm:
2 3 6
0
N -{[4-Amino-3-(3-bromobenzyl)-5-oxo-4,5-dihydro-1H-
164.8 (CO); 158.9 (C–O); 158.6 (C–O coumarin); 153.8 (C–O
1
,2,4-triazol-1-yl]acetyl}-6-chloro-2-oxo-2H-chromen-3-
triazole); 148.8 (CN); 146.6 (coumarin C-4); 145.6 (coumarin C-
carbohydrazide (8d)
Yield: 3.40 g (62% for microwave), 2.63 g (48% for
conventional), m.p.: 279–280°C, H NMR (DMSO-d
3); 133.4; 129.2; 128.7; 121.3; 121.1; 120.7 (Ar-C); 46.4 (NCH
2
);
3
5
35
35
þ
11.1 (CH
3
). LC-MS, m/z: 427.0419 [M (Cl )(Cl )þH] , 429.0391
1
37
37
þ
35
þ
6
), d, ppm:
[M (Cl )(Cl )þH] , 449.0258 [M (Cl )(Cl )þNa] , 451.0230
3
5
37
þ
37
37
þ
1
1.02 (1H, s, NH); 10.62 (1H, s, NH); 8.81 (1H, s, H-4); 8.11 (1H, d,
J¼ 2.4 Hz, Ar-H); 7.77 (1H, dd, J ¼ 8.8 Hz, J¼ 2.4 Hz, Ar-H); 7.53
1H, d, J ¼ 8.8 Hz, Ar-H); 7.46–7,40 (2H,m, Ar-H); 7.28–7.23 (2H, m,
[M (Cl )(Cl )þNa] , 453.3526 [M (Cl )(Cl )þNa] .
0
(
N -{[4-Amino-3-(4-chlorobenzyl)-5-oxo-4,5-dihydro-1H-
1
3
Ar-H);5.32(2H,s,NH
DMSO-d
), d, ppm: 164.6 (C –– O); 159.8 (CO); 158.8 (C –– O
coumarin); 153.8 (coumarin C-3); 152.9 (C –– O triazole); 147.1
coumarin C-3); 147.0 (C –– N); 139.0; 134.2; 131.9; 130.9; 130.0;
2
);4.45(2H, s, NCH
2
);3.90(2H, s, CH
2
). CAPT
1,2,4-triazol-1-yl]acetyl}-6,8-dichloro-2-oxo-2H-chromen-
3-carbohydrazide (8h)
(
6
Yield: 3.45 g (64% for microwave), 2.59 g (48% for
1
(
conventional), m.p.: 303–304°C, H NMR (DMSO-d
6
), d, ppm:
1
29.5; 129.3; 128.3; 122.0; 120.0; 119.7; 118.7 (Ar-C); 46.5 (NCH
2
);
10.99 (1H, s, NH); 10.59 (1H, s, NH); 8.79 (1H, s, H-4); 8.11 (1H, d,
35
79
þ
30.3(CH
2
). LC-MS, m/z:547.0187[M(Cl )(Br )þH] ,549.0171[M
J ¼ 2.4 Hz, Ar-H); 8.07 (1H, d, J ¼ 2.4 Hz, Ar-H); 7.30 (2H, d,
35
81
þ
37
81
þ
(Cl )(Br )þH] , 551.0141 [M (Cl )(Br )þH] .
J ¼ 6.4 Hz, Ar-H); 7.27 (2H, d, J ¼ 6.4 Hz, Ar-H); 5.30 (2H, s, NH
2
);
1
3
4
.44 (2H, s, NCH ); 3.88 (2H, s, CH ). C APT (DMSO-d ), d, ppm:
2 2 6
0
N -{[4-Amino-3-(2-bromobenzyl)-5-oxo-4,5-dihydro-1H-
,2,4-triazol-1-yl]acetyl}-6-chloro-2-oxo-2H-chromen-3-
164.7 (C–O); 158.8 (CO); 158.6 (C–O coumarin); 153.8 (C–O
triazole); 148.8 (C–N); 147.2 (coumarin C-3); 146.6 (coumarin
C-4); 135.2; 133.4; 131.8; 131.1; 129.2; 128.7; 128.6; 121.3;
1
carbohydrazide (8e)
Yield: 3.18 g (58% for microwave), 2.30 g (42% for
121.1; 120.7 (Ar-C); 46.5 (NCH
2
); 30.1 (CH
2
). LC-MS, m/z:
1
35
35
35
þ
37
35
conventional), m.p.: 258–259°C, H NMR (DMSO-d
6
), d, ppm:
537.0244 [M (Cl )(Cl )(Cl )þH] , 539.0221 [M (Cl )(Cl
)
3
5
þ
37
37
35
þ
1
0.99 (1H, s, NH); 10.60 (1H, s, NH); 8.80 (1H, s, H-4); 8.12 (1H, d,
(Cl )þH] , 541.0188 [M (Cl )(Cl )(Cl )þH] .
J ¼ 2.0 Hz, Ar-H); 7.77 (1H, dd, J ¼ 8.8 Hz, J ¼ 2.4 Hz, Ar-H);
0
7
NH
.60–7.53 (2H, m, Ar-H); 7.34–7.17 (3H,m, Ar-H); 5.34 (2H, s,
N -{[4-Amino-3-(4-bromobenzyl)-5-oxo-4,5-dihydro-1H-
13
2
); 4.43 (2H, s, NCH
2
); 3.99 (2H, s, CH
2
). C APT (DMSO-d
6
),
1,2,4-triazol-1-yl]acetyl}-6,8-dichloro-2-oxo-2H-chromen-
d, ppm: 164.6 (C–O); 159.8 (C–O); 158.8 (CO coumarin); 153.9
coumarin C-3); 153.0 (C–O triazole); 147.0 (coumarin C-4);
3-carbohydrazide (8i)
Yield: 3.71 g (64% for microwave), 1.86 g (32% for
conventional), m.p.: 299–300°C, H NMR (DMSO-d ), d, ppm:
6
(
1
1
1
46.4 (CN); 135.5; 134.2; 132.9; 131.4; 129.6; 129.4; 129.3;
28.3; 124.4; 120.1; 119.8; 118.7 (Ar-C); 46.5 (NCH ); 31.3 (CH ).
2
2
35
10.99 (1H, s, NH); 10.55 (1H, s, NH); 8.79 (1H, s, H-4); 8.11 (1H, d,
J ¼ 2.0 Hz, Ar-H); 8.07 (1H, d, J ¼ 2.0 Hz, Ar-H); 7.48 (2H, dd,
J ¼ 8.8 Hz, J ¼ 2.8 Hz, Ar-H); 7.20 (2H, dd, J ¼ 8.8 Hz, J ¼ 2.8 Hz,
35
79
þ
LC-MS, m/z: 547.0180 [M (Cl )(Br )þH] , 549.0161 [M (Cl
)
81
þ
37
81
þ
(
Br )þH] , 551.0127 [M (Cl )(Br )þH] .
2 2 2
Ar-H); 5.29 (2H, s, NH ); 4.44 (2H, s, NCH ); 3.86 (2H, s, CH ).
0
13
N -{[4-Amino-3-(3,4-dichlorobenzyl)-5-oxo-4,5-dihydro-
C APT (DMSO-d
6
), d, ppm: 165.7 (C–O); 164.7 (C–O); 158.9
1
3
H-1,2,4-triazol-1-yl]acetyl}-6-chloro-2-oxo-2H-chromen-
-carbohydrazide (8f)
(C–O coumarin); 153.9 (C–O triazole); 148.8 (C–N); 147.2
(coumarin C-3); 146.6 (coumarin C-4); 135.7; 133.4; 131.7;
131.5; 129.2; 128.7; 121.3; 121.2; 120.7; 120.3 (Ar-C); 46.5
Yield: 3.39 g (63% for microwave), 2.64g (49% for
1
79
35
35
conventional), m.p.: 289–290°C, H NMR (DMSO-d
1.02 (1H, s, NH); 10.61 (1H, s, NH); 8.80 (1H, s, H-4); 8.11 (1H, d,
J ¼ 2.8 Hz, Ar-H); 7.75 (1H, dd, J ¼ 8.8 Hz, J ¼ 2.4 Hz, Ar-H); 7.54–
6
), d, ppm:
(NCH
2
); 30.2 (CH
2
). LC-MS, m/z: 580.9700 [M (Br )(Cl )(Cl )þ
þ
81 35 35 þ 81
1
H] , 582.9681 [M (Br )(Cl )(Cl )þH] , 584.9653 [M (Br
)
3
7
35
þ
(Cl )(Cl )þH] .
7
.49 (3H, m, Ar-H); 7.24 (1H, dd, J ¼ 8.4 Hz, J ¼ 2,0 Hz, Ar-H); 5.32
13
0
(
2H, s, NH
2
); 4.46 (2H, s, NCH
2
); 3.91 (2H, s, CH
2
). C APT (DMSO-
N -{[4-Amino-3-(3-bromobenzyl)-5-oxo-4,5-dihydro-1H-
d
6
), d, ppm: 164.6 (C –– O); 159.8 (C –– O); 158.8 (C –– O coumarin);
1,2,4-triazol-1-yl]acetyl}-6,8-dichloro-2-oxo-2H-chromen-
1
1
1
53.9 (coumarin C-3); 152.9 (CO triazole); 147.0 (coumarin C-4);
46.8 (CN); 137.4; 134.2; 131.3; 131.2; 130.9; 129.8; 129.7; 129.5;
3-carbohydrazide (8j)
Yield: 3.89 g (67% for microwave), 3.19 g (55% for
conventional), m.p.: 302–303°C, H NMR (DMSO-d ), d, ppm:
6
11.00 (1H, s, NH); 10.56 (1H, s, NH); 8.79 (1H, s, H-4); 8.10 (1H, s,
1
29.3; 120.1; 119.6; 118.7 (Ar-C); 46.5 (NCH
2
); 29.9 (CH
2
). LC-MS,
3
5
35
35
þ
37
35
m/z: 537.0319 [M (Cl )(Cl )(Cl )þH] , 539.0292 [M (Cl )(Cl
)
35
þ
37
37
þ
(
Cl )þH] , 541.0263 [M (Cl )(Cl )þH] .
Ar-H); 8.06 (1H, s, Ar-H); 7.46–7.40 (2H, m, Ar-H); 7.27–7.20
(
2H, m, Ar-H); 5.32 (2H, s, NH
2
); 4.46 (2H, s, NCH
), d, ppm: 164.7 (C–O); 158.8 (CO);
2
); 3.88 (2H, s,
0
13
N -[(4-Amino-3-methyl-5-oxo-4,5-dihydro-1H-1,2,4-
triazol-1-yl)acetyl]-6,8-dichloro-2-oxo-2H-chromen-3-
carbohydrazide (8g)
Yield: 3.05 g (68% for microwave), 1.66 g (37% for
conventional), m.p.: 290–291°C, H NMR (DMSO-d
CH ). C APT (DMSO-d
2 6
158.6 (C–O coumarin); 153.8 (C–O triazole); 148.8 (C–N);
147.1 (coumarin C-3); 146.6 (coumarin C-4); 139.0; 133.5;
131.9; 130.9; 130.0; 129.2; 128.7; 128.4; 122.1; 121.3; 121.1;
1
6
), d, ppm:
120.7 (Ar-C); 46.6 (NCH
2
); 30.3 (CH
2
). LC-MS, m/z: 580.9766
ß 2017 Deutsche Pharmazeutische Gesellschaft
www.archpharm.com
(6 of 9) e1600369

Arch. Pharm. Chem. Life Sci. 2017, 350, e1600369
Coumarin–Triazole Hybrid Molecules
ARC HH PHARM
Archiv der Pharmazie
7
9
35
35
þ
81
35
[
5
M (Br )(Cl )(Cl )þH] , 582.9742 [M (Br )(Cl )(Cl35)þH]þ,
10.93 (1H, s, NH); 10.52 (1H, s, NH); 8.66 (1H, s, H-4); 7.70 (1H, d,
J ¼ 9.2 Hz, Ar-H); 7.35–7.27 (4H, m, Ar-H); 6.80 (1H, dd,
J ¼ 9.2 Hz, J ¼ 2.8 Hz, Ar-H); 6.60 (1H, d, J ¼ 2.4 Hz, Ar-H);
81
37
35
þ
84.9613 [M (Br )(Cl )(Cl )þH] .
0
N -{[4-Amino-3-(2-bromobenzyl)-5-oxo-4,5-dihydro-1H-
,2,4-triazol-1-yl]acetyl}-6,8-dichloro-2-oxo-2H-chromen-
-carbohydrazide (8k)
Yield: 2.79 g (48% for microwave), 1.80 g (31% for
5.30 (2H, s, NH
2
); 4.43 (2H, s, NCH
2
); 3.87 (2H, s, CH
2
); 3.47 (4H,
1
3
1
3
q, 2CH ); 1.10 (6H, t, 2CH
2
3
). C APT (DMSO-d ), d, ppm: 164.3
6
(C–O); 161.8 (C–O); 160.1 (CO coumarin); 157.8 (Ar-C); 153.8
(coumarin C-3); 153.2 (C–O triazole); 148.7 (coumarin C-4);
147.2 (CN); 135.3; 132.3; 131.8; 131.1 (2C); 128.8 (2C); 110.8;
1
conventional), m.p.: 286–287°C, H NMR (DMSO-d
6
), d, ppm:
1
0.99 (1H, s, NH); 10.56 (1H, s, NH); 8.78 (1H, s, H-4); 8.10 (1H, d,
2 2 2
108.1; 96.3 (Ar-C); 46.5 (NCH ); 44.8 (2CH ); 30.2 (CH ); 12.8
3
5
þ
35
J ¼ 2.0 Hz, Ar-H); 8.04 (1H, d, J ¼ 2.0 Hz, Ar-H); 7.58 (1H, d,
(2CH
3
). LC-MS, m/z: 540.35 [M (Cl )þH] , 562.38 [M (Cl )þ
þ
37 þ
J ¼ 7.6 Hz, Ar-H); 7.31–7.28 (2H, m, Ar-H); 7.20–7.17 (1H, m, Ar-
Na] ,564.33 [M (Cl )þNa] .
2 2 2
H); 5.32 (2H, s, NH ); 4.46 (2H, s, NCH ); 3.88 (2H, s, CH ).
13
0
C APT (DMSO-d
6
), d, ppm: 164.7 (C–O); 158.8 (C–O); 158.6
N -{[4-Amino-3-(4-bromobenzyl)-5-oxo-4,5-dihydro-1H-
1,2,4-triazol-1-yl]acetyl}-7-diethylamino-2-oxo-2H-
chromen-3-carbohydrazide (8o)
(
(
C–O coumarin); 153.9 (C–O triazole); 148.7 (C–N); 146.7
coumarin C-3); 146.5 (coumarin C-4); 135.5; 133.4; 132.9;
1
31.4; 129.4; 129.2; 128.6; 128.2; 124.4; 121.4; 121.2; 120.6 (Ar-
Yield: 3.27 g (56% for microwave), 1.93 g (33% for conven-
7
9
35
1
C); 46.6 (NCH
2
); 31.3 (CH
2
). LC-MS, m/z: 580.9760 [M (Br )(Cl
)
tional), m.p.: 259–260°C, H NMR (DMSO-d
6
), d, ppm: 10.93
35
þ
81
35
35
þ
(
(
Cl )þH] , 582.9745 [M (Br )(Cl )(Cl )þH] , 584.9715 [M
(1H, s, NH); 10.52 (1H, s, NH); 8.67 (1H, s, H-4); 7.70 (1H, d,
J ¼ 7.2 Hz, Ar-H); 7.48 (2H, d, J ¼ 8.8 Hz, Ar-H); 7.23 (2H, d,
J ¼ 8.8 Hz, Ar-H); 6.80 (1H, dd, J ¼ 8.0 Hz, J ¼ 2.0 Hz, Ar-H); 6.61
81
37
35
þ
Br )(Cl )(Cl )þH] .
0
N -{[4-Amino-3-(3,4-dichlorobenzyl)-5-oxo-4,5-dihydro-
1
chromen-3-carbohydrazide (8l)
Yield: 2.74 g (48% for microwave), 2.63g (46% for
(1H, d, J ¼ 2.0 Hz, Ar-H); 5.29 (2H, s, NH
2
); 4.43 (2H, s, NCH
); 3.47 (4H, q, 2CH ); 1.14 (6H, t, 2CH
2 2 3
2
);
). C APT
), d, ppm: 164.3 (C–O); 161.8 (C–O); 160.1 (C–O
6
13
H-1,2,4-triazol-1-yl]acetyl}-6,8-dichloro-2-oxo-2H-
3.85 (2H, s, CH
(DMSO-d
coumarin); 157.8 (Ar-C); 153.9 (coumarin C-3); 153.2 (C–O
triazole); 148.7 (coumarin C-4); 147.1 (CN); 135.7; 132.3; 131.7;
1
conventional), m.p.: 310–311°C, H NMR (DMSO-d
6
), d, ppm:
1
1.01 (1H, s, NH); 10.56 (1H, s, NH); 8.79 (1H, s, H-4); 8.10 (1H, d,
J ¼ 2.4 Hz, Ar-H); 8.04 (1H, d, J ¼ 2.0 Hz, Ar-H); 7.54–7.51 (2H, m,
Ar-H); 7.26 (1H, d, J ¼ 9.4 Hz, Ar-H); 5.32 (2H, s, NH ); 4.46 (2H, s,
), d, ppm: 164.6 (CO);
58.8 (C –– O); 158.6 (C –– O coumarin); 153.9 (CO triazole); 148.7
C –– N); 146.8 (coumarin C-3); 146.6 (coumarin C-4); 137.4; 133.4;
31.3;131.2;131.1;130.9;129.8;129.2;128.7;121.3;121.1;120.7
2 2
131.5; 120.3; 110.8; 108.1; 96.3 (Ar-C); 46.5 (NCH ); 44.8 (2CH );
79
þ
30.3 (CH
586.1257 [M (Br )þH] .
2
); 12.8 (2CH
3
). LC-MS, m/z: 584.1273 [M (Br )þH] ,
8
1
þ
2
13
2 2 6
NCH ); 3.91 (2H, s, CH ). C APT (DMSO-d
1
(
1
0
N -{[4-Amino-3-(3-bromobenzyl)-5-oxo-4,5-dihydro-1H-
1,2,4-triazol-1-yl]acetyl}-7-diethylamino-2-oxo-2H-
chromen-3-carbohydrazide (8p)
3
5
(
(
5
Ar-C); 46.6 (NCH
2
); 29.9 (CH
2
). LC-MS, m/z: 570.9900 [M (Cl
)
Yield: 3.79 g (65% for microwave), 1.87 g (32% for conven-
Cl )(Cl )(Cl )þH] , 572.9872 [M (Cl )(Cl )(Cl )(Cl )þH]þ,
35
35
35
þ
37
35
35
35
tional), m.p.: 257–258°C, H NMR (DMSO-d
1
6
), d, ppm: 11.00
37
37
35
35
þ
74.9841 [M (Cl )(Cl )(Cl )(Cl )þH] .
(1H, s, NH); 10.59 (1H, s, NH); 8.74 (1H, s, H-4); 7.77 (1H, d,
J ¼ 6.8 Hz, Ar-H); 7.53 (1H, s, Ar-H); 7.49–7.47 (1H, m, Ar-H);
7.33–7.31 (2H, m, Ar-H); 6.88 (1H, d, J ¼ 6.8 Hz, Ar-H); 6.74 (1H,
0
N -[(4-Amino-3-methyl-5-oxo-4,5-dihydro-1H-1,2,4-
triazol-1-yl)acetyl]-7-diethylamino-2-oxo-2H-chromen-3-
carbohydrazide (8m)
s, Ar-H); 5.39 (2H, s, NH
3.54 (4H, q, 2CH
2
); 4.45 (2H, s, NCH
2
); 3.96 (2H, s, CH
2
);
1
3
2
); 1.19 (6H, t, 2CH
3
). C APT (DMSO-d ), d,
6
Yield: 2.23 g (52% for microwave), 1.50 g (35% for
ppm: 164.3 (C–O); 161.8 (C–O); 160.1 (C–O coumarin); 157.8
(Ar-C); 153.9 (coumarin C-3); 153.2 (CO triazole); 148.7 (C–N);
147.0 (coumarin C-4); 139.0; 132.3; 131.9; 130.9; 130.0; 128.3;
1
conventional), m.p.: 246–247°C, H NMR (DMSO-d
6
), d, ppm:
1
0.91 (1H, s, NH); 10.51 (1H, s, NH); 8.67 (1H, s, H-4); 7.71 (1H, d,
J ¼ 8.8 Hz, Ar-H); 6.81 (1H, d, J ¼ 8.0 Hz, Ar-H); 6.61 (1H, s, Ar-
H); 5.25 (2H, s, NH ); 4.39 (2H, s, NCH ); 3.84 (4H, q, 2CH ); 2.09
3H, s, CH ); 1.12 (6H, t, 2CH ). C APT (DMSO-d ), d, ppm:
64.4 (C–O); 161.8 (C–O); 160.1 (CO coumarin); 157.8 (Ar-C);
120.0; 110.8; 108.1; 96.4 (Ar-C); 46.5 (NCH
2
); 44.8 (2CH
2
); 30.3
7
9
þ
2
2
2
(CH
2
); 12.8 (2CH
3
). LC-MS, m/z: 584.1319 [M (Br )þH] ,
13
81
þ
(
3
3
6
586.1305 [M (Br )þH] .
1
1
4
0
53.8 (coumarin C-3); 153.3 (CO triazole); 148.7 (coumarin C-
); 145.6 (CN); 132.3; 110.8; 108.1; 108.0; 96.4 (Ar-C); 46.3; 44.8
N -{[4-Amino-3-(2-bromobenzyl)-5-oxo-4,5-dihydro-1H-
1,2,4-triazol-1-yl]acetyl}-7-diethylamino-2-oxo-2H-
chromen-3-carbohydrazide (8q)
þ
(
2CH
2
); 12.8 (2CH
3
); 11.1. LC-MS, m/z: 430.1878 [MþH] ,
þ
4
52.1728 [MþNa] .
Yield: 2.80 g (48% for microwave), 1.63 g (28% for conven-
1
tional), m.p.: 260–261°C, H NMR (DMSO-d
6
), d, ppm: 11.91
0
N -{[4-Amino-3-(4-chlorobenzyl)-5-oxo-4,5-dihydro-1H-
(1H, s, NH); 10.52 (1H, s, NH); 8.66 (1H, s, H-4); 7.69 (1H, d,
J ¼ 8.8 Hz, Ar-H); 7.60 (1H, d, J ¼ 8.8 Hz, Ar-H); 7.29–7.18 (4H,
m, Ar-H); 6.80 (1H, d, J ¼ 7.6 Hz, Ar-H); 6.59 (1H, s, Ar-H); 5.33
1
,2,4-triazol-1-yl]acetyl}-7-diethylamino-2-oxo-2H-
chromen-3-carbohydrazide (8n)
Yield: 3.45 g (64% for microwave), 1.99 g (37% for
(2H, s, NH
2 2 2
); 4.43 (2H, s, NCH ); 3.99 (2H, s, CH ); 3.46 (4H, q,
1
13
conventional), m.p.: 263–264°C, H NMR (DMSO-d
6
), d, ppm:
2CH ); 1.11 (6H, t, 2CH
2
3
). C APT (DMSO-d
6
), d, ppm: 164.2
ß 2017 Deutsche Pharmazeutische Gesellschaft
www.archpharm.com
(7 of 9) e1600369

Arch. Pharm. Chem. Life Sci. 2017, 350, e1600369
B. Kahveci et al.
ARC HH PHARM
Archiv der Pharmazie
(
(
(
C–O); 161.8 (C–O); 160.1 (C–O coumarin); 157.8 (Ar-C); 153.9
coumarin C-3); 153.2 (C–O triazole); 148.7 (C–N); 146.4
coumarin C-4); 135.5; 132.9; 132.2; 131.4; 129.3; 128.2; 124.3;
Lipase inhibition assay
The inhibitory effects of those compounds were evaluated
against porcine pancreatic lipase (Applichem, Germany)
(15 ng/mL). Lipase activity assay was done according to
Kurihara et al. [39]. The lipase activity was measured using
4-methylumbelliferyl oleate (4-MU oleate) as a substrate.
Briefly, compounds were mixed with porcine pancreatic
lipase (PPL) 1:3 (v/v) and incubated for 30 min. The
microtiter plates containing 50 mL 0.1 mM 4-MU oleate,
1
1
10.8; 108.1; 96.4 (Ar-C); 46.5 (NCH
2
); 44.8 (2CH
2
); 31.3 (CH
2
);
). LC-MS, m/z: 584.1324 [M (Br )þH] , 586.1309 [M
þ
79
þ
2.8 (2CH
3
81
(
Br )þH] .
0
N -{[4-Amino-3-(3,4-dichlorobenzyl)-5-oxo-4,5-dihydro-
1
H-1,2,4-triazol-1-yl]acetyl}-7-diethylamino-2-oxo-2H-
chromen-3-carbohydrazide (8r)
25 mL diluted compound-lipase solution, 25 mL dH
assay buffer (13 mM Tris-HCl, 150 mM NaCl, and 1.3 mM
CaCl , pH 8.0) were incubated at 37°C for 20 min. After
2
O and
Yield: 3.40 g (59% for microwave), 2.19 g (38% for
1
conventional), m.p.: 286–287°C, H NMR (DMSO-d
6
), d, ppm:
2
1
1.92 (1H, s, NH); 10.50 (1H, s, NH); 8.67 (1H, s, H-4); 7.71 (1H, d,
incubation, in order to stop the reaction, 0.1 mL 0.1 M pH
4.2 citrate buffer was added to the reaction mixture. The
amount of 4-methylumbelliferone released by the lipase
was measured by using a spectrofluorometer (SpectraMax
M5, Molecular Devices) at an excitation wavelength of
355 nm and an emission wavelength of 460 nm. The
inhibitory activity of those compounds and orlistat
(Xenical, Hoffman, La Roche, Segrate, Italy), an inhibitor
control of pancreatic lipase, were measured at various
concentrations. Residual activities were calculated by
comparing to control without inhibitor. The assays were
done in triplicate. The IC50 value was determined as the
concentration of compound that gives 50% inhibition of
maximal activity.
J ¼ 9.2 Hz, Ar-H); 7.56–7.51 (2H, d, Ar-H); 7.25 (1H, dd,
J ¼ 8.0 Hz, J ¼ 1.6 Hz, Ar-H); 6.82 (1H, d, J ¼ 8.8 Hz, Ar-H);
6
.61 (1H, s, Ar-H); 5.29 (2H, s, NH
2
); 4.43 (2H, s, NCH
2
); 3.91 (2H,
). C APT (DMSO-
), d, ppm: 164.3 (C–O); 161.8 (CO); 160.2 (C–O coumarin);
57.9 (Ar-C); 153.8 (coumarin C-3); 153.3 (CO triazole); 148.7
coumarin C-4); 146.8 (C–N); 137.4; 132.3; 131.4; 131.2; 130.9;
13
2 2 3
s, CH ); 3.48 (4H, q, 2CH ); 1.11 (6H, t, 2CH
d
1
(
6
1
2
2 2
29.8; 129.7; 110.8; 108.1; 96.4 (Ar-C); 46.5 (NCH ); 44.8 (2CH );
35
35
9.8 (CH
2
); 12.8 (2CH
3
). LC-MS, m/z: 574.1347 [M (Cl )(Cl )þ
þ
37 35 þ
H] , 576.1320 [M (Cl )(Cl )þH] .
Biological activity
Cytotoxic activity assay
Antitumor activities of compounds 8a–r in vitro were
determined colorimetrically using the tetrazolium salt
This work was supported by The Scientific and Technological
Research Council of Turkey (TUBITAK) under the project
number of 115Z047. Authors thank to TUBITAK for this
support.
(
MTT) assay. Tumor cells (BT20 human breast carcinoma,
SK-Mel 128 melanoma, DU-145 prostate carcinoma, and A549
lung carcinoma) were grown in RPMI 1640 (Lonza, USA)
medium supplemented with 10% fetal calf serum (FCS) and
gentamycin/fungizon. Human fibroblast cells were grown in
The authors have declared no conflicts of interest.
5
a-MEM (Lonza, USA). Cells (1 ꢁ 10 per mL) were seeded into a
9
6-well plate, then the compounds studied were added in a
concentration gradient, and the final concentrations were
maintained at 100, 31.6, 10, 3.16, and 1 mg/mL, respectively.
The plate was maintained at 37°C in a humidified atmosphere
References
[1] R. Shah, J. Singh, D. Singh, A. S. Jaggi, N. Singh, Eur. J.
Med. Chem. 2016, 114, 170–190.
2
of 5% CO and incubated for 48 h. Antineoplastic cisplatin
was used as positive control. After incubation, media were
[2] L. Jia, S. T. Ma, Eur. J. Med. Chem. 2016, 121, 209–220.
replaced on 100 mL fresh RPMI-1640 1% FCS and antibiotics for
[3] National
breast
cancer
reports.
http://www.
3
0 min at 37°C in a humidified atmosphere of 5% CO
2
. Then,
nationalbreastcancer.org/what-is-cancer, 2016 (Accessed
11.08.2016).
1
0 mL MTT (3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyltetrazo-
lium bromide) solution of an 5 mg/mL concentration was
added to each well. After incubation for 3 h at 37°C, media
were removed and 100 mL DMSO was added to all wells for
solubilization of formazan crystal. Measurement of the
absorbance of the solutions, related to the number of live
cells, was carried out using a microplate reader (Tecan,
Sunrise) at 570 nm. Cell cytotoxicity (CC50) values were
calculated by GraphPad Prism software using dose–response
curves. The SI is defined as the ratio of the CC50 obtained from
the experiments on normal cells to that obtained from cancer
cells. As the SI demonstrates the differential activity of the
compound, the greater the SI value is, the more selective the
compound is [37, 38].
[4] World Health Organization report. http://www.who.int/
topics/obesity/en/, 2016 (Accessed 11.08.2016).
[5] M. Al-Bachir, I. Othman, Soc. Sci. J. 2016, 53, 184–189.
[6] R. Sahani, S. Chakrabarty, P. Bharati, Obes. Res. Clin.
Pract. 2010, 4, E119–E125.
[7] J. Dandriyal, R. Singla, M. Kumar, V. Jaitak, Eur. J. Med.
Chem. 2016, 119, 141–168.
[8] J. Bronikowska, E. Szliszka, D. Jaworska, Z. P. Czuba,
W. Krol, Molecules 2012, 17, 6449–6464.
[9] A. A. H. Kadhum, A. A. Al-Amiery, A. Y. Musa,
A. B. Mohamad, Int. J. Mol. Sci. 2011, 12, 5747–5761.
[10] S. Weigt, N. Huebler, R. Strecker, T. Braunbeck,
T. H. Broschard, Reprod. Toxicol. 2012, 33, 133–141.
ß 2017 Deutsche Pharmazeutische Gesellschaft
www.archpharm.com
(8 of 9) e1600369

Arch. Pharm. Chem. Life Sci. 2017, 350, e1600369
Coumarin–Triazole Hybrid Molecules
ARC HH PHARM
Archiv der Pharmazie
[
[
[
[
[
[
11] S. M. de Souza, F. Delle Monache, A. Smania, Z.
[26] P. L. Zhao, W. F. Ma, A. N. Duan, M. Zou, Y. C. Yan,
W. W. You, S. G. Wu, Eur. J. Med. Chem. 2012, 54,
813–822.
[27] F. T. Silva, C. H. Franco, D. C. Favaro, L. H. Freitas,
C. B. Moraes, E. I. Ferreira, Eur. J. Med. Chem. 2016, 121,
553–560.
[28] B. Kahveci, Molecules 2005, 10, 376–382.
[29] B. Kahveci, F. Yilmaz, E. Mentese, S. Ulker, Chem.
Heterocycl. Com. 2015, 51, 447–456.
[30] V. V. Kouznetsov, A. Gomez-Barrio, Eur. J. Med. Chem.
2009, 44, 3091–3113.
Naturforsch. C 2005, 60, 693–700.
12] R. V. Patel, P. Kumari, D. P. Rajani, K. H. Chikhalia, Med.
Chem. Res. 2013, 22, 195–210.
13] S. Sardari, Y. Mori, K. Horita, R. G. Micetich, S. Nishibe,
M. Daneshtalab, Bioorgan. Med. Chem.1999,7,1933–1940.
14] Y. Bansal, P. Sethi, G. Bansal, Med. Chem. Res. 2013, 22,
3
049–3060.
15] A. Thakur, R. Singla, V. Jaitak, Eur. J. Med. Chem. 2015,
01, 476–495.
1
16] R. R. Zhang, J. Liu, Y. Zhang, M. Q. Hou, M. Z. Zhang,
F. E. Zhou, W. H. Zhang, Eur. J. Med. Chem. 2016, 116,
[31] M. H. Shaikh, D. D. Subhedar, V. M. Khedkar, P. C. Jha,
F. A. K. Khan, J. N. Sangshetti, B. B. Shingate, Chinese
Chem. Lett. 2016, 27, 1058–1063.
76–83.
[
17] R. Nagamallu, B. Srinivasan, M. B. Ningappa,
A. K. Kariyappa, Bioorg. Med. Chem. Lett. 2016, 26,
[32] E. Mentese, F. Yilmaz, F. Mutlu, B. Kahveci, J. Chem. Res.
2015, 39, 645–648.
[33] M. C. Alley, D. A. Scudiero, A. Monks, M. L. Hursey,
M. J. Czerwinski, D. L. Fine, B. J. Abbott, J. G. Mayo,
R. H. Shoemaker, M. R. Boyd, Cancer Res. 1988, 48,
589–601.
6
90–694.
18] F. Wen, H. Jin, K. Tao, T. P. Hou, Eur. J. Med. Chem. 2016,
20, 244–251.
19] T. Nasr, S. Bondock, M. Youns, Eur. J. Med. Chem. 2014,
6, 539–548.
[
[
[
1
7
20] N. Kulabas, E. Tatar, O. B. Ozakpinar, D. Ozsavci,
C. Pannecouque, E. De Clercq, I. Kucukguzel, Eur. J.
Med. Chem. 2016, 121, 58–70.
[34] A. A. Vandeloosdrecht, R. H. J. Beelen, G. J. Ossenkoppele,
M. G. Broekhoven, M. M. A. C. Langenhuijsen, J. Immunol.
Methods 1994, 174, 311–320.
[
21] D. Gupta, D. K. Jain, J. Adv. Pharm. Technol. 2015, 6,
[35] D. A. Scudiero, R. H. Shoemaker, K. D. Paull, A. Monks,
S. Tierney, T. H. Nofziger, M. J. Currens, D. Seniff,
M. R. Boyd, Cancer Res. 1988, 48, 4827–4833.
[36] A. R. Katritzky, A. Abdelmajeid, S. R. Tala, M. S. Amine,
P. J. Steel, Synthesis-Stuttgart 2011, 1, 83–90.
[37] P. Prayong, S. Barusrux, N. Weerapreeyakul, Fitoterapia
2008, 79, 598–601.
[38] A. Koch, P. Tamez, J. Pezzuto, D. Soejarto, J. Ethno-
pharmacol. 2005, 101, 95–99.
[39] H. Kurihara, S. Asami, H. Shibata, H. Fukami, T. Tanaka,
Biol. Pharm. Bull. 2003, 26, 383–385.
141–146.
[
22] R. Paprocka, M. Wiese, A. Eljaszewicz, A. Helmin-Basa,
A. Gzella, B. Modzelewska-Banachiewicz, J. Michalkiewicz,
Bioorg. Med. Chem. Lett. 2015, 25, 2664–2667.
23] R. Kaur, A. R. Dwivedi, B. Kumar, V. Kumar, Anti-Cancer
Agent Med. Chem. 2016, 16, 465–489.
24] S. G. Kucukguzel, P. Cikla-Suzgun, Eur. J. Med. Chem.
[
[
[
2015, 97, 830–870.
25] K. Sztanke, T. Tuzimski, J. Rzymowska, K. Pasternak,
M. Kandefer-Szerszen, Eur. J. Med. Chem. 2008, 43, 404–419.
ß 2017 Deutsche Pharmazeutische Gesellschaft
www.archpharm.com
(9 of 9) e1600369