Synthesis of Some New Pyrazoline-Based Thiazole Derivatives and Evaluation of Their Antimicrobial, Antifungal, and Anticancer Activities

Article abstract of DOI:10.1134/S1068162020030061

Abstract: 3-(2-Thienyl)-5-aryl-1-thiocarbamoyl-2-pyrazolines were reacted with chloroacetone derivatives and hydrazonyl chloride derivatives in ethanol to afford the corresponding thiazolylpyrazoline derivatives and thiophenylpyrazolyl-5-substituted aryl-

Full text of DOI:10.1134/S1068162020030061

ISSN 1068-1620, Russian Journal of Bioorganic Chemistry, 2020, Vol. 46, No. 3, pp. 382–392. © Pleiades Publishing, Ltd., 2020.
Synthesis of Some New Pyrazoline-Based Thiazole Derivatives
and Evaluation of Their Antimicrobial, Antifungal,
and Anticancer Activities
Safaa I. Elewaa, Eman Mansoura, Ibrahim F. Nassarb, 1, and Amal A. I. Mekaweyc
aDepartment of Chemistry, Faculty of Women for Arts, Science and Education, Egypt, Ain Shams University,
Cairo, 11566 Egypt
bFaculty of Specific Education, Ain Shams University, Abassia, Cairo, 11566 Egypt
cThe Regional Center of Mycology and Biotechnology, Al-Azhar University, Cairo, 11651 Egypt
Received May 27, 2019; revised July 3, 2019; accepted December 20, 2019
Abstract—3-(2-Thienyl)-5-aryl-1-thiocarbamoyl-2-pyrazolines were reacted with chloroacetone deriva-
tives and hydrazonyl chloride derivatives in ethanol to afford the corresponding thiazolylpyrazoline deriv-
atives and thiophenylpyrazolyl-5-substituted aryl-diazenylthiazole derivatives, respectively. The structures
of the newly synthesized compounds were elucidated by different elemental and spectral analyses (IR,
mass, 1H and 13C NMR). The antimicrobial and antifungal activities of the newly synthesized compounds
were evaluated against four bacterial species and five fungal strains. In addition, the antitumor activities of
two of the newly synthesized compounds 1-(2-(5-(4-chlorophenyl)-3-(thiophen-2-yl)-4,5-dihydropyra-
zol-1-yl)-4-methyl thiazol-5-yl)ethan-1-one and 2-(5-(4-chlorophenyl)-3-(thiophen-2-yl)-4,5-dihydro-
1H-pyrazol-1-yl)-4-methyl-5-(phenyl-diazenyl)thiazole against HEPG-2, HCT-116, MCF-7, BHK, and
CACO-2 were evaluated. From the obtained results, we found that these two compounds were the most
potent candidates towards all gram-positive and gram-negative bacteria, as well as the fungi studied. Also,
the same two compounds showed strong antitumor activities against two of the tumor cell lines (HCT-116
and CACO-2).
Keywords: thiophene, thiazole, pyrazoline, antimicrobial, antifungal, anticancer activity
DOI: 10.1134/S1068162020030061
INTRODUCTION
receptor tyrosine kinase (EGFR-TK) inhibitors [17],
aurora kinase inhibitors [18], COX-2/B-Raf inhibitors
[19], telomerase inhibitors [20], and tubulin assem-
bling inhibitors [21]. On the other hand, thiazole has
been recognized as a promising scaffold for the design
and development for central nervous system (CNS)
active agents. There are thiazole-based CNS drugs
currently used as therapeutic agents for the treatment
of various CNS disorders and a number of thiazole
derivatives are in clinical trials [22]. Diverse modifica-
tions of the thiazole ring at various positions have led
to a variety of thiazole-based CNS agents as AChE
and BuChE inhibitors, secretase inhibitors, mono-
amine oxidase (MAO) inhibitors, neuronal nitric
oxide synthase inhibitors, Ach receptor ligands, ade-
nosine receptor ligands, dopamine receptor ligands,
serotonin receptor ligands, glutamate receptor ligands,
γ-aminobutyric acid receptor ligands, opioid receptor
ligands, neuroprotective, and anticonvulsant agents
[22–25]. Acotiamide has been reported to be a prom-
The presence of pyrazole nucleus in different
structures leads to diversified applications in different
areas, such as technology, medicine, and agriculture.
In particular, they are described as inhibitors of pro-
tein glycation, antibacterial, antifungal, anti-tubercu-
losis, antioxidant, and antiviral agents [1, 2]. Com-
pounds including a pyrazole nucleus exhibit a wide
spectrum of biological activities, such as antimicro-
bial, anti-inflammatory, antidepressant, and antican-
cer effects. Among the reported activities, it is import-
ant to note that pyrazolines are not only useful in
treatment of various cancer types, including brain,
bone, mouth, esophagus, stomach, liver, bladder,
pancreas, cervix, lung, breast, colon, rectum, and
prostate cancers, but also some of them act as cancer
chemopreventive agents [3–21]. Also, pyrazoline
derivatives were reported as epidermal growth factor
1 Corresponding
sedu.asu.edu.eg.
author:
e-mail:
Dr.Ibrahim.Nassar@
382

SYNTHESIS OF SOME NEW PYRAZOLINE-BASED THIAZOLE DERIVATIVES
383
ising thiazole-based agent for the treatment of func- bands of C=O, C=N, and C=C groups at their
1
tional dyspepsia in clinical trials. Acotiamide
enhances ACh release in the enteric nervous system
through AChE inhibition and M1/M2 muscarinic
receptor antagonism [26]. Many studies were carried
out on heterocyclic systems bearing thiazole and pyr-
azoline [27–32] and pyrazolidinone [33] groups as
pharmacophores. Sulfur and/or nitrogen heterocy-
cles that possess pharmaceutical activities widely
occur in nature in the form of alkaloids, vitamins,
pigments, and as constituents of plant and animal
cells. Thiazolyl-pyrazoline derivatives have been
used as cholinesterase inhibitors [34]. From the
above facts and our interest in designing new hetero-
cyclic compounds that have the pharmaceutical
properties [35–39], we synthesized a new system that
combines these two bio-labile components, pyrazo-
lines and thiazoles, together in order to give a com-
pact structure like the titled compounds. Evaluation
of the antimicrobial and antifungal activities of the
newly synthesized compounds was investigated
against four bacterial species and/or five fungal
strains. In addition, the antitumor activities of two
synthesized compounds against HEPG-2, HCT-116,
MCF-7, BHK, and CACO-2 were also evaluated.
expected regions. H NMR spectra of the same
compounds are consistent with their assigned struc-
1
tures. The H NMR spectra of the compounds
(VIIa–i) showed CH2 signals of the pyrazoline ring
resonating as a pair of doublets of doublets at δ
3.15–3.70 (Ha) and 3.98–4.16 ppm (Hb). The (Hc)
proton appeared as a doublet of doublets at δ 5.59–
5.90 ppm due to vicinal coupling with the two mag-
netically non-equivalent protons of the methylene
group at position 4 of the pyrazoline ring (Ja–b
=
18.50–23.10 Hz, Ja–c = 6.0–6.80 Hz, Jb–c = 11.70–
12.10 Hz). The other aromatic and aliphatic protons
were found at their expected regions. Also,
13C NMR spectral data of the same compounds
gave another evidence for the assigned structures
(Experimental part and Scheme 1).
On the other hand, reaction of 3-(2-thienyl)-5-
aryl-1-thiocarbamoyl-2-pyrazoline derivative (IIIa)
(R = H) with hydrazonyl chloride derivatives (VIIIa–c)
(R2 = a; H, b; 4-Me and c; 4-Cl) afforded 3-(thio-
phen-2-yl)-4,5-dihydropyrazol-1-yl)-diazenylthiazol
derivatives (XIa–c), respectively. Also, reaction of
compound (IIIb) (R = Cl) with compounds (VIIIa–c)
afforded compounds (XId–f), respectively. Finely,
reaction of compound (IIIc) (R = OMe) with hydra-
zonyl chloride derivatives (VIIIa–c) afforded com-
pounds (XIg–i), respectively. All the reactions were
performed in absolute ethanol under reflux tempera-
ture. The reaction mechanism involves two intermedi-
ates (IXa–i) and (Xa–i) that played important roles in
thiazole formation step. The structures of compounds
(XIa–i) were confirmed by elemental and spectral
analyses, including GC-MS, IR, H, and C NMR.
Their 1H NMR spectra showed the appearance of the
characteristic signals due to the three pyrazoline ring
protons in the regions of δ 3.33–3.37, 3.90–4.15, and
5.73–5.97 ppm, respectively, due to vicinal coupling
with the two magnetically non-equivalent protons of
the methylene group at position 4 of the pyrazoline
RESULTS AND DISCUSSION
Chemistry
3-(2-Thienyl)-5-aryl-1-thiocarbamoyl-2-pyra-
zolines (IIIa–c) (R = a, H; b, Cl; c, OMe) were pre-
pared by the reaction of 1-(2-thienyl)-3-aryl-2-
propen-1-ones (Ia–c) (R = a, H; b, Cl; c, OMe)
with thiosemicarbazide and sodium hydroxide in
ethanol according to a reported procedure [40]. The
reaction mechanism involves the formation of
hydrazone followed by subsequent addition of N–H
on the olefinic bond of the propenone moiety [41].
Reaction of the thioamide derivative (IIIa) (R = H)
with chloroacetone derivatives (IVa–c) (R1 = a;
Me, b; OEt and c; NHPh) yielded thiophenyl-pyra-
zolyl-thiazoles (VIIa–c), respectively, as shown in
Scheme 1. The reactions were performed in reflux-
ing ethanol. Reaction of compound (IIIb) (R = Cl)
with compounds (IVa–c) under the same condi-
tions afforded compounds (VIId–f), respectively.
Finely, reaction of (IIIc) (R = OMe) with com-
pounds (IVa–c) at the same conditions afforded
compounds (VIIg–i), respectively. The mecha-
nisms of all reactions passed through two interme-
diates (Va–i) and (VIa–i), respectively. The struc-
tures of compounds (VIIa–i) were confirmed by
elemental and spectral analyses using methods of
GC-MS, IR, 1H, and 13C NMR. IR spectra of com-
1
13
ring (Ja–b = 18.50–23.10 Hz, Ja–c = 6.0–6.80 Hz, Jb–c
=
11.70–12.10 Hz). All other aromatic and aliphatic pro-
tons were found at their expected regions. The IR
spectra of compounds (XIa–i) revealed absorption
bands at about 1600–1605 cm–1 due to the C=N
groups. 13C NMR spectral data of the compounds give
us another proof for the same assigned structures (see
pounds (VIIa–i) showed stretching absorption the Experimental section and Scheme 2).
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 46 No. 3 2020

384
SAFAA I. ELEWA et al.
H2N
N
S
O
S
N
NaOH, EtOH
H2N
+
N
H
NH2
R
S
Ref.
R
S
(II)
(Ia−c); R = a, H
b, Cl
(IIIa−c); R = a, H
b, Cl
c, OMe
c, OMe
O O
R1
Cl
EtOH, Ref.
(IVa−c)
R1 = a, Me
b, OEt
c, NHPh
R1
R1
HO
N
O
HN
R1
N
N
O
O
H
S
S
O
S
N
N
N
Hc
N
N
R
R
R
Ha
Hc
S
S
S
(VIIa−i)
(VIa−i)
(Va−i)
(V), (VI), (VII)
R
R1
Me
OEt
NHPh
Me
Yield, %
a
b
c
d
e
f
g
h
i
H
H
H
Cl
Cl
Cl
56
76
67
77
56
68
65
78
72
OEt
NHPh
OMe Me
OMe OEt
OMe NHPh
Scheme 1. Synthesis of pyrazoline-based thiazoles (VIIa–i).
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY
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SYNTHESIS OF SOME NEW PYRAZOLINE-BASED THIAZOLE DERIVATIVES
385
HN
R2
H2N
O
HN
S
N
R2
N
O
N
S
EtOH
Ref.
N
+
N
N
N
R
H
Cl
S
R
S
(IIIa−c)
(VIIIa−c)
R2 = a, H
b, Me
(IXa−i)
c, Cl
R2
R2
N
N
N
N
H
HO
N
−H2O
N
S
S
N
N
N
N
R
R
S
S
(XIa−i)
(Xa−i)
R2
(IX), (X), (XI)
R
Yield, %
a
b
c
d
e
f
g
h
i
H
H
H
Cl
Cl
Cl
H
66
66
86
76
90
90
66
90
66
Me
Cl
H
Me
Cl
H
OMe
OMe Me
OMe Cl
Scheme 2. Synthesis of pyrazoline-based thiazoles (XIa–i).
Antimicrobial Activity
part of the results was those that were obtained from
antifungal activity screening. Most of the compounds
were effective against C. albicans and A. fumigatus
when compared with griseofulvin, especially com-
pounds (VIIb) and (XId), which showed strong activi-
ties. Meanwhile, compounds (VIIe), (VIIa), (VIId),
(VIIg), (VIIf), (Xia), (XId), and (VIIc) showed mod-
erate activities against the same strains compared with
griseofulvin. In the case of thiazole derivatives, potent
antifungal activities were observed comparable with a
commercially available drug, in particular against
The antimicrobial activity of the newly synthesized
compounds was evaluated against two gram-positive
bacteria, Staphylococcus aureus (S. a.) and Bacillus
cereus (B. c.), and two gram-negative bacteria, Klebsi-
ella pneumonia and Pseudomonas aeruginosa,
together with to five different fungus strains, namely,
Candida albicans, Syncephalastrum racemosum,
Aspergillus fumigatus, Penicillium expansum, and
Aspergillus flavus.
The results of the antimicrobial screening of the C. albicans and A. fumigatus. Antibacterial activity
newly synthesized pyrazole derivatives (VIIa–i) and assessment revealed that the compounds had moder-
(XIa–i) are shown in Table 1. The most important ate or slight activities.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 46 No. 3 2020

386
SAFAA I. ELEWA et al.
Table 1. Antibacterial and antifungal activities of the synthesized compounds
Inhibition zone, mm
Comp.
(5 mg/mL)
Antibacterial activity
Antifungal activity
S. aureus B. subtilis K. pneumonia P. aeruginosa A. fumigatus A. flavus S. racemosum P. expansum C. albicans
(VIIb)
(VIIe)
(VIIh)
(VIIa)
(VIId)
(VIIg)
(VIIc)
(VIIf)
(VIIi)
(Xia)
2.2
1.1
–
–
1.5
–
–
1.5
–
–
1.8
–
1
0.5
–
0.9
–
–
–
0.5
–
–
0.8
–
0.2
1.1
–
0.9
–
–
–
–
–
–
–
–
1
–
–
0.9
–
1
20
–
–
–
–
0.5
0.5
–
0.5
1.1
–
–
–
–
0.5
2
1.8
–
–
–
–
–
–
–
–
0.8
1.2
–
0.6
–
1.2
0.9
–
–
0.5
–
–
–
–
–
0.9
1
–
0.8
–
1.2
–
–
0.8
0.5
–
1.2
–
1
1
–
0.8
–
1
–
1.4
–
–
1.9
–
–
–
0.5
2.2
–
1.1
–
(XId)
(XIg)
(XIb)
(XIe)
1.5
–
–
1.5
–
0.6
–
1
–
–
–
Std.a
2.3
2.4
2.8
3.5
2.2
3.3
2.4
2.1
3
a Antibacterial reference drug is amoxicilline and antifungal reference drug is griseofulvin.
Minimum inhibitory concentration (MIC). By cell line. Compound (VIIb) did not show any cytotoxic
applying ethyl acetate solvent of the synthesized com- effect on the BHK normal fibroblast cell line, so com-
pounds started with a maximum concentration of pound (VIIb) may be considered as a better candidate
500 mg/mL and then reduced it by successive two fold antitumor agent than compound (XId).
dilutions of the stock solution using a calibrated
micropipette. MIC of the sample determination was
Structure–Activity Relationship (SAR)
carried out by inoculation of their serial dilutions with
test organisms and measurement of inhibition zones
using diffusion agar technique. MIC was expressed as
the lowest concentration inhibiting test. The results of
minimum inhibitory concentration showed that, pyr-
azole derivatives (VIIb) and (XId) (MIC 1.25 mg/ mL)
showed better results than the reference drug (MIC
2.5 mg/mL).
From the results of the antibacterial and antifungal
activities we concluded that compounds (VIIb) and
(XId) were the most potent towards almost all the
gram-positive and gram-negative bacteria, as well as
all fungi, studied. Also, the two compounds, (VIIb)
and (XId), showed strong antitumor activities against
two tumor cell lines (HCT-116 and CACO-2). This
may be due to the C=S, C=N, C=O, and N=N
groups found in the heterocyclic rings, which gave
good binding energies between the compounds and
the active sites found in the proteins, in addition to the
aromatic rings and the ester group found in compound
(VIIb) and the halide atom (–Cl) found in compound
(XId), which increased the activity of such com-
pounds. The other compounds showed moderate
effects toward the studied bacteria and fungus.
Antitumor Activity
The antitumor activity of the two synthesized com-
pounds (VIIb) and (XId) was evaluated against five
tumor cell lines (HEPG-2 liver carcinoma cell line,
MCF-7 breast carcinoma cell line, HCT-116 colon
carcinoma cell line, CACO-2 colon carcinoma cell
line, and BHK normal fibroblast cell line). The results
of the antitumor activity of the newly synthesized
compounds against the five tumor cell lines (survival, %)
are summarized in Table 2, which shows the IC50 val-
ues, i.e. the doses that reduces survival to 50%. From
the results we conclude that compounds (VIIb) and tives (VIIa–i) and (XIa–i) were synthesized and eval-
(XId) have strong activities against the HCT-116 and uated for their antibacterial, antifungal, and antitumor
CACO-2 lines while only compound (XId) shows high activities. A significant level of activity was illustrated.
cytotoxic affect against the BHK normal fibroblast Compounds (VIIb) and (XId) were the most potent
CONCLUSION
Thiazolyl-3-(2-thienyl)-5-aryl-2-pyrazoline deriva-
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SYNTHESIS OF SOME NEW PYRAZOLINE-BASED THIAZOLE DERIVATIVES
387
Table 2. Antitumor activities of compounds (VIIb) and (XId) against five tumor cell lines (survival, %, and IC50, µg/mL)
Cell survival, %
Concentration, µg/mL
(VIIb)
(XId)
Concentration, µg/mL
(VIIb)
(XId)
0.0
100
70
100
96
92
87
83
–
0.0
100
86
77
62
48
8.9
100
95
83
70
59
–
100
80
71
1.0
1.0
2.5
55
2.5
5.0
43
5.0
47
10.0
30
10.0
34
IC50, µg/mL
IC50, µg/mL
100
100
100
95
4.0
0.0
100
98
94
86
77
–
0.0
100
87
63
58
38
1.0
1.0
2.5
2.5
5.0
84
5.0
10.0
70
10.0
IC50, µg/mL
–
IC50, µg/mL
5.9
0.0
1.0
2.5
5.0
100
89
100
90
85
58
44
6.0
75
58
10.0
IC50, µg/mL
40
7.5
towards almost all the gram-positive and gram-nega- d, doublet; q, quartet; m, multiplet. The mass spectra
tive bacteria, as well as all fungi, studied. Also, the two were recorded on a Shimadzu GC-MS QP-1000 EX
compounds (VIIb) and (XId) showed a strong antitu- mass spectrometer operating at 70 eV. The progress of
mor activity against two tumor cell lines (HCT-116 all reactions was monitored by thin layer chromatogra-
and CACO-2). Only compound (XId) showed high phy using Merck Kiesel gel 60-F254 aluminum backed
cytotoxic affect against the BHK cells. Compound plates. Unless otherwise noted, all solvents and
(VIIb) did not show any cytotoxic effect on the BHK reagents were commercially available and used with-
normal fibroblast cell line, so we concluded that com- out further purification. Compounds (IIIa–c) were
pound (VIIb) may be considered as a better candidate synthesized according to a reported procedure [40].
antitumor drug than compound (XId).
For compound (IIIa), mp 198–200°; (IIIb), mp 198–
200°C; (IIIc), mp 144–146°C. Hydrazonoyl halides
were prepared as reported in the literature [45].
EXPERIMENTAL
Chemistry
General Procedure for Synthesis of Compounds (VIIa–i)
and (XIa–i)
Melting points were measured with a Gallenkamp
melting point apparatus and were uncorrected. Infra-
red (IR) spectra (ν, cm–1), were recorded as KBr disks
using the Perkin Elmer FT-IR Spectrum BX appara-
tus. Mass spectral data are given as m/z (intensity, %).
NMR spectra were recorded on a Bruker NMR spec-
trometer. 1H NMR spectra were recorded at 400 MHz
and 13C spectra, at 100 MHz, in deuterated dimethyl-
sulfoxide (DMSO-d6). Chemical shifts are expressed
To a suspension of compounds (IIIa–c) (10 mmol;
(IIIa), 2.87 g; (IIIb), 3.21 g; (IIIc), 3.17 g) in absolute
ethanol (30 mL), compounds (IVa–c) or (VIIIa–c)
(10 mmol; α-chloro acetyl acetone (IVa), 1.29 g; α-
chloroethyl acetoacetate (IVb), 1.64 g; α-chloroaceto-
acetanilide (IVc), 2.11 g; α-chloroaceto phenylhydra-
zono, (Z)-1-chloro-1-(2-phenylhydrazono)propan-
2-one (VIIIa), 1.96 g, (Z)-1-chloro-1-(2-p-tolylhy-
in δ values (ppm) using the solvent peak as internal drazono)propan-2-one (VIIIb), 2.10 g; and (Z)-1-
standard. All coupling constants (J) values are given in chloro-1-(2-(4-chlorophenyl) hydrazono)propan-2-
hertz. The abbreviations used are as follows: s, singlet; one (VIIIc), 2.30 g) were added and heated at reflux
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 46 No. 3 2020

388
SAFAA I. ELEWA et al.
1-(2-(5-(4-Chlorophenyl)-3-(thiophen-2-yl)-4,5-
temperature for 8 h. After cooling, the precipitates
were collected. The products were crystallized from dihydropyrazol-1-yl)-4-methylthiazol-5-yl)ethan-1-one
ethanol to afford the titled compounds (VIIa–i) and (VIId). Yield 3.09 g (77%); dark green crystals; mp
(XIa–i), respectively.
219–220°C; IR: 1660 (C=O), 1606 (C=N), 1575
1
(C=C); H NMR: 2.37 (s, 3H, CH3), 2.39 (s, 3H,
1-(4-Methyl-2-(5-phenyl-3-(thiophen-2-yl)-4,5-
COCH3), 3.35 (m, 1H, pyrazoline Ha), 4.05 (dd, J =
12.4, J = 11.2 Hz, 1H, pyrazoline Hb), 5.77 (dd, J =
12.4, J = 5.2 Hz, 1H, pyrazole Hc), 7.16–7.49 (m, 4H,
thiophene H4, H5, Ar-H),7.49–7.50 (d, J = 1.2 Hz,
1H, thiophene H3),7.79–7.80 (m, 2H, Ar-H); MS (m/z):
dihydropyrazol-1-yl)thiazol-5-yl)ethanone
(VIIa).
Yield 2.05 g (56%); dark green crystals; mp 202–
204°C; IR: 3084 (Ar-CH), 2929 (aliphatic CH), 1625
1
(C=O); H NMR: 2.38 (s, 3H, CH3), 2.52 (s, 3H,
COCH3 ), 3.33 (dd, 1H, J = 11.6, J = 5.2 Hz, pyrazo-
line Ha), 4.04–4.12 (dd, 1H, J = 18.4, J = 11.6 Hz,
pyrazoline Hb), 5.64–5.70 (dd, 1H, J = 9.6, J =
5.2 Hz, pyrazol Hc), 7.16–7.30 (m, 3H, Ar-H),7.34–
7.38 (m, 3H, thiophene-H4, Ar-H ),7.49–7.50 (d, 1H,
J = 2.8 Hz, thiophene-H5), 7.78–7.79 (d, 1H, J = 0.8
402 ([M+ + 1], 5); analysis calcd. for C19H16ClN3OS2:
C, 56.78; H, 4.01; N, 10.45; found: C, 56.63; H, 4.22;
N, 10.34%.
Ethyl
2-(5-(4-chlorophenyl)-3-(thiophen-2-yl)-
4,5-dihydropyrazol-1-yl)-4-methylthiazole-5-carbox-
ylate (VIIe). Yield 2.41 g (56%); pale green crystals;
mp 160–162°C; IR: 1678 (C=O), 1579 (C=N);
1H NMR: 1.26 (t, J = 6, 3H, OCH2CH3), 3.35 (s, 3H,
CH3), 3.34 (m, 1H, pyrazoline Ha), 4.03–4.11 (dd, J =
17.6, J = 10.4 Hz, 1H, pyrazoline Hb), 4.16–4.22 (q,
J = 8.4 Hz, 2H, OCH2CH3), 5.73–5.77 (dd, J = 12,
J = 6.4 Hz, 1H, pyrazol Hc),7.17–7.31 (m, 3H, thio-
phene H4, Ar-H),7.40–7.49 (m, 3H, thiophene H5,
Ar-H), 7.79–7.80 (d, J = 3.2 Hz, 1H, thiophene H3);
Hz, thiophene-H3); 13C NMR: 16.0, 26.4 (2CH3),
44.66 (CH2), 63.73 (CH), 124.4, 128.10, 128.98,
129.28, 130.36, 130.98, 134.07, 136.0, 138.3 139.55,
141.66, 150.79, 155.27, 160.87 (Ar-C, thiophene-C,
thiazole-C, pyrazole-C), 196.35 (CO); MS (m/z): 369
([M+ + 2], 5), 367 ([M+], 12); analysis calcd. for
C19H17N3OS2: C, 62.10; H, 4.66; N, 11.43; found: C,
62.21; H, 4.11; N, 10.50%.
Ethyl-4-methyl-2-(5-phenyl-3-(thiophen-2-yl)-4,5-
dihydropyrazol-1-yl)thiazole-5-carboxylate
13C NMR: 14.73, 17.88 (2CH3), 44.49, 60.69 (2CH2),
63,07 (CH), 111.03, 125.6, 126.0, 127.6, 128.69,
129.28, 130.74, 131.35, 132.69, 133.74, 140.40, 151.66,
159.57, (Ar-C, thiophene-C, thiazole-C, pyrazole-
C), 162.23 (CO),164.92 (CN); MS (m/z): 432 ([M+ +
1], 6); analysis calcd. for C20H18ClN3O2S2: C, 55.61;
H, 4.20; N, 9.73; found: C, 55.41; H, 4.33; N, 10.0%.
(VIIb).
Yield 3.02 g (76%); bright green crystals; mp 144–
145°C; IR: 1735 (C=O), 1618 (C=N), 1595 (C=C);
1H NMR: 1.28 (t, 3H, J = 7.2 Hz, OCH2CH3), 2.35 (s,
3H, CH3), 3.39 (m, 1H, pyrazoline Ha), 4.04–4.12
(dd, J = 18.4, J =12 Hz, 1H, pyrazoline Hb), 4.16–
4.42 (q, J = 7.2 Hz, 2H, OCH2CH3), 5.72–5.77 (dd,
J = 13.2, J = 6 Hz, 1H, pyrazol Hc), 7.16–7.38 (m, 6H,
H4 thiophene, Ar-H),7.49–7.50 (d, J = 3.6 Hz, 1H,
H5 thiophene), 7.78–7.79 (d, J = 2.8 Hz, 1H, H3-thio-
2-(5-(4-Chlorophenyl)-3-(thiophen-2-yl)-4,5-
dihydropyrazol-1-yl)-4-methyl-N-phenyl-thiazole-5-
carboxamide (VIIf). Yield 3.25 g (68%); bright green
powder; mp 218–220°C; IR: 1641 (C=O), 1596
phene); MS (m/z): 397 ([M+], 5); analysis calcd. for
C20H19N3O2S2: C, 60.43; H, 4.82; N, 10.57; found: C,
60.11; H, 4.77; N, 10.40.
1
(C=N); H NMR: 2.34 (s, 3H, CH3), 3.37 (m, 1H,
pyrazoline Ha), 4.04–4.12 (dd, J = 18.8, J = 14 Hz,
1H, pyrazoline Hb), 5.73–5.78 (dd, J = 14.8, J =
5.6 Hz, 1H, pyrazol Hc), 7.05–7.32 (m, 8H, thio-
phene H4, Ar-H),7.43–7.48 (m, 2H, Ar-H), 7.63–
7.65 (d, J = 6.8 Hz, 1H, thiophene H5), 7.76–7.78 (d,
J = 5.2 Hz, 1H, thiophene H3), 9.66 (s, 1H, NH
4-Methyl-N-phenyl-2-(5-phenyl-3-(thiophen-2-yl)-
4,5-dihydropyrazol-1-yl)thiazole-5-carboxamide (VIIc).
Yield 2.98 (67%); pale yellow crystals; mp 232–
233°C; IR: 1570–1595 (C=C); 1H NMR: 2.37 (s, 3H,
CH3), 3.34–3.39 (m, 1H, pyrazoline Ha), 4.05–4.13
(dd, J = 19.2, J = 9.6 Hz, 1H, pyrazoline Hb), 5.74–
5.78 (dd, J = 12.4, J =7. 6 Hz, 1H, pyrazol Hc),7.05–
7.48 (m, 11H, thiophene H4, Ar-H),7.76–7.78 (d, J =
4.8 Hz, 1H, thiophene H5),7.63–7.65 (d, J = 2.8 Hz,
1H, thiophene H3), 9.67 (s, 1H, NH exchangeable);
exchangeable); MS (m/z): 480 ([M+ + 1], 11); analysis
calcd. for C24H19ClN4OS2: C, 60.18; H, 4.00; N, 11.70;
found: C, 60.11; H, 4.07; N, 11.77%.
1-(2-(5-(4-Methoxyphenyl)-3-(thiophen-2-yl)-4,5-
dihydropyrazol-1-yl)-4-methylthiazol-5-yl)ethanone (VIIg).
Yield 2.58 g (65%); greenish silver crystals; mp 198–
200°C; IR: 1640 (C=O), 1579 (C=C); 1H NMR: 2.38
(s, 3H, thiazole CH3), 2.50 (s, 3H, COCH3), 3.40 (m,
1H, pyrazoline Ha), 3.72 (s, 3H, OCH3), 4.02 (dd, J =
21, J = 12.4 Hz, 1H, pyrazoline Hb), 5.69 (dd, J = 11.6,
J = 4.8 Hz, 1H, pyrazole Hc), 6.87–6.92 (d, J = 8 Hz,
2H, Ar-H), 7.17–7.19 (m, 3H, thiophene H4, Ar-H),
7.30 (d, J = 5.6 Hz, 1H, thiophene H5), 7.77 (d, J = 6 Hz,
13C NMR: 18.14 (CH3), 44.66 (CH2), 63.73 (CH),
115.55, 120.83, 121.0, 123.90, 124.4, 125.0, 126.28,
128.10, 128.98, 129.28, 130.36, 130.98, 134.07, 136.0,
138.3 139.55, 141.66, 150.79, 155.27, 160.87 (Ar-C,
thiophene-C, thiazole-C, pyrazole-C), 163.35 (CO);
MS (m/z): 444 ([M+], 4); analysis calcd. for
C24H20N4OS2: C, 64.84; H, 4.53; N, 12.60; found: C,
64.61; H, 4.73; N, 12.90%.
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SYNTHESIS OF SOME NEW PYRAZOLINE-BASED THIAZOLE DERIVATIVES
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1H, thiophene H3); 13C NMR: 19.07, 30.08 (2CH3),
40.40 (CH2), 55.5 (CH3), 63.2 (CH), 114.5, 115.55,
128.10, 128.98, 129.28, 130.98, 134.07, 136.0, 138.3
139.55, 141.66, 150.79, 155.27, 160.87 (Ar-C, thio-
phene-C, thiazole-C, pyrazole-C), 189.4 (CO); MS
(m/z): 497 ([M+], 15); analysis calcd. for
C20H19N3O2S2: C, 60.43; H, 4.82; N, 10.57; found: C,
60.11; H, 4.75; N, 10.90%.
4-Methyl-2-(5-phenyl-3-(thiophen-2-yl)-4,5-dihydro-
1H-pyrazol-1-yl)-5-(p-tolyldiazenyl)-thiazole (XIb).
Yield 2.927 g (66%); scarlet red crystals; mp 194–
196°C; IR: 1572 (C=N), 1595 (C=C); 1H NMR: 2.36
(s, 3H, Ar-CH3), 2.48 (s, 3H, thiazole CH3), 3.91–
3.98 (dd, J = 18.8, J = 13.2 Hz, 1H, pyrazoline Ha),
4.08–4.13 (dd, J = 21.2, J = 12.8 Hz, 1H, pyrazoline
Hb), 5.85–5.89 (dd, J = 13.6, J = 9.2 Hz, 1H, pyrazol
Hc), 7.14–7.16 (m, 6H, Ar-H, thiophene H4,
H5),7.14–7.83 (m, 6H Ar-H, thiophene H3); MS
Ethyl 2-(5-(4-methoxyphenyl)-3-(thiophen-2-yl)-
4,5-dihydropyrazol-1-yl)-4-methylthiazole-5-carbox-
ylate (VIIh). Yield 3.33 g (78%); pale yellow crystals;
mp 152–153°C; IR: 1735 (C=O), 1578 (C=C);
1H NMR: 1.26 (t, J = 6.4 Hz, 3H, OCH2CH3 ), 2.35
(s, 3H, thiazole CH3), 3.29–3.30 (m, 1H, pyrazoline
Ha), 3.72 (s, 3H, OCH3), 4.02 (dd, J = 19.6, J =
12.8 Hz, 1H, pyrazoline Hb), 4.19 (q, J = 9.6 Hz, 2H,
OCH2CH3), 5.66 (dd, J = 13.2, J = 6.0 Hz, 1H, pyra-
zole Hc), 6.90 (m, 2H, Ar-H), 7.12–7.19 (m, 3H, thio-
phene H4, Ar-H) , 7.48 (d, J = 2.8 Hz, 1H, thiophene
H5), 7.78 (d, J = 6.8 Hz, 1H, thiophene H3); MS
(m/z): 443 ([M+], 12); analysis calcd. for C24H21N5S2:
C, 64.98; H, 4.77; N, 15.79; found: C, 64.81; H, 4.67;
N, 15.90%.
5-((4-Chlorophenyl)diazenyl)-4-methyl-2-(5-phe-
nyl-3-(thiophen-2-yl)-4,5-dihydro-1H-pyrazol-1-yl)-
thiazole (XIc). Yield 3.99 g (86%); reddish brown crys-
tals; mp 190–192°C; IR: 1572 (C=N); 1H NMR: 2.48
(s, 3H, thiazole CH3), 3.38–3.44 (m, 1H, pyrazoline
Ha), 3.89–3.98 (dd, J = 21.2, J = 12.8 Hz, 1H, pyra-
zoline Hb), 5.87–5.91 (dd, J = 12.8, J = 2.8 Hz, 1H,
pyrazole Hc),7.16–7.85 (m, 12H, Ar-H, thiophene H3,
(m/z): 427 ([M+], 22); analysis calcd. for
C21H21N3O3S2: C, 59.00; H, 4.95; N, 9.83; found C,
58.95; H, 4.90; N, 9.80%.
H4, H5); MS (m/z): 465 ([M+ + 1], 22); analysis calcd.
for C23H18ClN5S2: C, 59.54; H, 3.91; N, 15.09; found:
C, 59.85; H, 3.55; N, 15.10%.
2-(5-(4-Methoxyphenyl)-3-(thiophen-2-yl)-4,5-
dihydropyrazol-1-yl)-4-methyl-N-phenyl-thiazole-5-
carboxamide (VIIi). Yield 3.42 g (72%); pale yellow
crystals; mp 224–225°C; IR: 1634 (C=O), 1615
(C=N), 1593 (C=C); 1H NMR: 2.36 (s, 3H, thiazole
CH3), 3.77 (m, 1H, pyrazoline Ha), 3.72 (s, 3H,
OCH3), 4.06 (dd, J = 23.6, J = 14 Hz, 1H, pyrazoline
Hb), 5.68 (dd, J = 13.2, J = 6 Hz, 1H, pyrazole Hc),
6.91 (d, J = 8 Hz, 2H, Ar-H), 7.05–7.50 (m, 8H, Ar-H,
thiophene H4), 7.63–7.65 (d, J = 6.8 Hz, 1H, thio-
phene H3), 7.76–7.74 (d, J = 6.8 Hz, 1H, thiophene
H5), 9.6 (s, 1H, NH exchangeable); MS (m/z): 476
2-(5-(4-Chlorophenyl)-3-(thiophen-2-yl)-4,5-dihydro-
1H-pyrazol-1-yl)-4-methyl-5-(phenyldiazenyl)thiazole
(XId). Yield 3.526 g (76%); orange crystals; mp 170–
1
172°C; IR: 1595 (C=N); H NMR: 2.51 (s, 3H, thi-
azole CH3), 3.13–3.18 (dd, 1H, J = 20, J = 4.0 Hz,
pyrazoline Ha), 3.91–3.98 (dd, J = 20, J = 12.0 Hz,
1H, pyrazoline Hb), 5.93–5.97 (dd, J =12, J = 4 Hz,
1H, pyrazole Hc), 7.16–7.55 (m, 7H, Ar-H, thiophene
H4, H5), 7.69–7.71 (m, 5H, thiophene H3, Ar-H); MS
(m/z): 465 ([M+ + 1], 27); analysis calcd. for C23H18-
ClN5S2: C, 59.54; H, 3.91; N, 15.09; found: C, 59.61;
H, 4.01; N, 15.0%.
([M+ + 2], 8); analysis calcd. for C25H22N4O2S2: C,
63.27; H, 4.67; N, 11.81; found: C, 63.41; H, 4.53; N,
11.90%.
2-(5-(4-Chlorophenyl)-3-(thiophen-2-yl)-4,5-dihydro-
1H-pyrazol-1-yl)-4-methyl-5-(p-tolyldiazenyl)thiazole
(XIe). Yield 4.30 g (90%); scarlet red crystals; mp
1
148–150°C; IR: 1618 (CN), 1595 (C=C); H NMR:
4-Methyl-2-(5-phenyl-3-(thiophen-2-yl)-4,5-dihydro-
1H-pyrazol-1-yl)-5-(phenyldiazenyl)-thiazole (XIa).
Yield 2.40 g (66%); orange crystals; mp 160–162°C;
IR: 1572 (C=N); 1H NMR: 2.51 (s, 3H, CH3), 3.91–
3.97 (dd, J = 16.4, J =10.8 Hz 1H, pyrazoline Ha),
4.01–4.17 (dd, J = 16, J =10 Hz, 1H, pyrazoline Hb),
2.27, 2.49 (2s, 6H, 2CH3), 3.14–3.29 (dd, 1H, J = 16,
J = 12 Hz, pyrazoline Ha), 3.91–3.98 (dd, J = 16,
J =12. Hz, 1H, pyrazoline Hb), 5.94–5.97 (dd, J =
11.6, J = 4. 8 Hz, 1H, pyrazole Hc), 7.15–7.17 (m, 6H,
Ar-H, H4, thiophene H5) 7.34–7.5 (m, 5H, thiophene
5.86–5.91 (dd, J = 14.8, J = 9.6 Hz, 1H, pyrazol Hc), H3, Ar-H); 13C NMR: 10.07, 21.0 (2CH3), 40.40
7.16–7.58 (m, 2H, H4, thiophene H5), 7.68–7.84 (m, (CH2), 60.2 (CH), 115.55, 120.83, 121.0, 123.90,
11H, Ar-H, thiophene H3); 13C NMR: 10.07 (CH3),
40.40 (CH2), 60.2 (CH), 124.60, 125.0, 126.28, 128.10,
128.98, 129.28, 130.36, 134.07, 135.2, 136.0, 138.3
139.55, 141.66, 147, 150.79, 155.27, 160.87, 163.35
164.2, 165.8 (Ar-C, thiophene-C, thiazole-C, pyra-
zole-C); MS (m/z): 429 ([M+], 10); analysis calcd. for
C23H19N5S2: C, 64.31; H, 4.46; N, 16.30; found: C,
64.11; H, 4.55; N, 16.50%.
124.4, 125.0, 126.28, 128.10, 128.98, 129.28, 130.36,
134.07, 136.0, 138.3 139.55, 141.66, 150.79, 155.27,
160.87, 163.35 (Ar-C, thiophene-C, thiazole-C, pyra-
zole-C), MS (m/z): 479 ([M+ + 1], 23); analysis calcd.
for C24H20ClN5S2: C, 60.30; H, 4.22; N, 14.65; found:
C, 60.11; H, 4.33; N, 14.50%.
2-(5-(4-Chlorophenyl)-3-(thiophen-2-yl)-4,5-dihydro-
1H-pyrazol-1-yl)-5-((4-chlorophenyl)diazenyl)-4-
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SAFAA I. ELEWA et al.
methylthiazole (XIf). Yield 4.486 g (90%); brownish
red crystals; mp 260–262°C; IR: 1625–1685 (C=N),
Antimicrobial Activities
Microorganisms. Nine clinical strains employed
for this investigation include four filamentous fungi
(Aspergillus fumigatus, Aspergillus flavus, Syncephlasta-
rum racemosum, and Penicillium expansum), one yeast
(Candida albicans), two gram-positive (Staphylococcus
aureus and Bacillus subtilis) and two gram-negative
(Klebsilla pnumonia and Pseudomonas aeruginosa)
bacteria. All strains were provided from culture collec-
tion of Regional Center for Mycology and Biotechnol-
ogy (RCMB), Al-Azhar University, Cairo, Egypt.
Antimicrobial assays. By diffusion agar technique,
the antifungal and antibacterial potentialities against
the tested species were expressed as the measurement
of diameter of their inhibition zone. Hole-plate diffu-
sion method was used; six equidistant (1 cm diameter)
holes were made using sterile cork borer in malt
extract agar and nutrient agar sterile plates (10 × 10
cm), which had previously been seeded with tested
fungal and bacterial isolates, respectively. Holes were
filled with 100 mL of three concentrations 5, 2.5, and
1 mg/mL of each of the synthesized compounds after
completely dissolving in ethyl acetate. Control holes
were filled with ethyl acetate solvent.
1
1570–1595 (C=C); H NMR: 2.5 (s, 3H, thiazole
CH3), 3.13–3.19 (dd, 1H, J = 16, J = 5.2 Hz, pyrazo-
line Ha), 3.90–3.98 (dd, J = 17.6, J = 10.8Hz, 1H,
pyrazoline Hb), 5.92–5.96 (dd, J = 16.8, J = 5.2 Hz,
1H, pyrazole Hc), 7.15–7.61 (m, 10H, Ar-H, thio-
phene H4, H5), 7.78–7.79 (d, J = 4.4 Hz, 1H, thio-
phene H3); MS (m/z): 499 ([M+ + 1], 12); analysis
calcd. for C23H17Cl2N5S2: C, 55.42; H, 3.44; N, 14.05;
found: C, 55.61; H, 3.77; N, 14.10%.
2-(5-(4-Methoxyphenyl)-3-(thiophen-2-yl)-4,5-
dihydro-1H-pyrazol-1-yl)-4-methyl-5-(phenyldiaze-
nyl)thiazole (XIg). Yield 3.033 g (66%); reddish brown
crystals; mp 90–92°C; IR: 1605 (C=N),1578 (C=C);
1H NMR: 2.5 (s, 3H, thiazole CH3), 3.74 (s,
3H,OCH3), 3.34–3.72 (m, 1H, pyrazoline Ha), 4.05–
4.12 (dd, J = 16, J = 8 Hz, 1H, pyrazoline Hb), 5.78–
5.87 (dd, J = 13.6, J = 11.6 Hz, 1H, pyrazole Hc),
6.87–7.84 (m, 12H, Ar-H, thiophene H3, H4, H5);
MS (m/z): 459 ([M+], 32); analysis calcd. for
C24H21N5OS2: C, 62.72; H, 4.61; N, 15.24; found: C,
62.51; H, 4.77; N, 15.40%.
Plates were left in a cooled incubator at 4 2°C for
1 h, then incubated at 37 2°C for bacterial isolates
and incubated at 28 2°C for fungal isolates. Inhibi-
2-(5-(4-Methoxyphenyl)-3-(thiophen-2-yl)-4,5-
dihydro-1H-pyrazol-1-yl)-4-methyl-5-(p-tolyldiaze- tion zones developed due to active ingredients were mea-
sured after 24–48 h of incubation time. Griseofulvin was
used as a standard antifungal agent while, amoxicilline
was used as a standard antibacterial agent [42].
nyl)thiazole (XIh). Yield 4.262 g (90%); brown crystals;
mp 200–202°C; IR: 1612 (C=N), 1580 (C=C);
1H NMR: 2.36, 2.49 (2s, 6H, 2CH3), 3.35 (m, 1H,
pyrazoline Ha), 3.73 (s, 3H, OCH3), 4.04–4.11 (dd,
J = 19.2, J = 10.8 Hz, 1H, pyrazoline Hb), 5.79–5.83
(dd, J = 11.6, J = 3.6 Hz, 1H, pyrazole Hc), 6.92–6.94
(m, 2H, Ar-H), 7.18–7.58 (m, 8H, thiophene H4, H5,
Ar-H), 7.83–7.82 (d, J = 5.6 Hz, 1H, thiophene H3);
Minimum inhibitory concentration (MIC) assays.
Determination of MIC was performed by a serial dilu-
tion technique [43]. Appling ethyl acetate solvent of
the synthesized compounds starting with the maxi-
mum concentration of 500 mg/mL, which then was
reduced by successive two-fold dilutions of the stock
solution using a calibrated micropipette. MIC of the
sample was determined by inoculation of the serial
dilutions with tested organisms and measurement of
inhibition zones using diffusion agar technique. MIC
was expressed as the lowest concentration inhibiting
test organism’s growth [44].
MS (m/z): 474 ([M+ + 1], 42); analysis calcd. for
C25H23N5OS2: C, 63.40; H, 4.90; N, 14.79; found: C,
63.61; H, 4.65; N, 14.90%.
5-((4-Chlorophenyl)diazenyl)-2-(5-(4-methoxy-
phenyl)-3-(thiophen-2-yl)-4,5-dihydro-1H-pyrazol-1-
yl)-4-methylthiazole (XIi). Yield 3.26 g (66%); reddish
brown crystals; mp 193–195°C; IR: 1610 (C=N), 1588
Antitumor Activity
1
(C=C); H NMR: 2.51 (s, 3H, CH3), 3.74 (s, 3H,
Five tumor cell lines (the HEPG2 liver carcinoma
cell line, MCF7 breast carcinoma cell line; HCT116
colon carcinoma cell line, and CACO colon carci-
noma cell line) and one normal cell line (BHK normal
fibroblast cell line) were provided from the National
Cancer Institute (NCI), Cairo University.
Cytotoxicity by SRB assay. Potential antitumor
activity and cytotoxicity of the compounds were tested
using the SRB technique. Tumor cells were plated in
96-well plate (104 cells/well) for 24 h before treatment
with compounds to allow attachment of cell to the wall
of the plate. Then, different concentrations of each
OCH3), 3.68–3.81 (m, 1H, pyrazoline Ha), 4.22–4.32
(m, 1H, pyrazoline Hb), 5.40–5.50 (m, 1H, pyrazol
Hc), 6.91–7.77 (m, 11H, Ar-H, thiophene H-3,4,5);
13C NMR: 10.07 (CH3), 40.40 (CH2), 55.0 (OCH3),
60.2 (CH), 115.55, 120.83, 121.0, 123.90, 124.4, 125.0,
126.28, 128.10, 128.98, 129.28, 130.98, 134.07, 136.0,
138.3 139.55, 141.66, 150.79, 155.27, 160.87, 163.35
(Ar-C, thiophene-C, thiazole-C, pyrazole-C); MS
(m/z): 494 ([M+], 34); analysis calcd. for C24H20-
ClN5OS2: C, 58.35; H, 4.08; N, 14.18; found: C,
58.21; H, 4.07; N, 14.20%.
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14. Montoya, A., Quiroga, J., Abonia, R., Nogueras, M.,
Cobo, J., and Insuasty, B., Molecules, 2014, vol. 19,
pp. 18 656–18 675.
compound (0, 1, 2.5, 5, and 10 µg/mL) were added to
the cell monolayer triplicate wells. Monolayer cells
were incubated with the compound for 48 h at 37°C
and in atmosphere of 5% CO2. After 48 h, cells were
fixed, washed, and stained with sulfo-rhodamine B
stain (SRB). Excess of stain was washed with acetic
acid and attached stain was recovered with Tris–
EDTA buffer. The color intensity was measured in an
ELISA reader. The relation between surviving fraction
and drug concentration is plotted to get the survival
curve of each tumor cell line.
https://doi.org/10.3390/ molecules191118656
15. Insuasty, B., Montoya, A., Becerra, D., Quiroga, J.,
Abonia, R., Robledo, S., Darío Vélez, I., Upegui, Y.,
Nogueras, M., and Cobo, J., Eur. J. Med. Chem., 2013,
vol. 67, pp. 252–262.
16. Rathore, P., Yaseen, S., Ovais, S., Bashir, R., Yaseen, R.,
and Hameed, A.D., Samim, M., Gupta, R., Hussain, F.,
and Javed, K., Bioorg. Med. Chem. Lett., 2014, vol. 24,
pp. 1685–1691.
17. Lv, P.-C., Li, D.-D., Li, Q.-S., Lu, X., Xiao, Z.-P., and
Zhu, H.-L., Bioorg. Med. Chem. Lett., 2011, vol. 21,
pp. 5374–5377.
18. Shin, S.Y., Yoon, H., Hwang, D.; Ahn, S., Kim, D.-W.,
Koh, D., Lee, Y.H., and Lim, Y., Bioorg. Med. Chem.,
2013, vol. 21, pp. 7018–7024.
COMPLIANCE WITH ETHICAL STANDARDS
The work has no studies involving humans or animals as
subjects of the study.
19. Yu, M., Yang, H., Wu, K., Ji, Y., Ju, L., and Lu, X.,
Conflict of Interests
Bioorg. Med. Chem., 2014, vol. 22, pp. 4109–4118.
Authors declare they have no conflicts of interest.
20. Amin, K.M., Abou-Seri, S.M., Awadallah, F.M., Eis-
sa, A.A.M., Hassan, G.S., and Abdulla, M.M., Eur. J.
Med. Chem., 2015, vol. 90, pp. 221–231.
21. Qin, Y.-J., Li, Y.-J., Jiang, A.-Q., Yang, M.-R.,
Zhu, Q.-Z., Dong, H., and Zhu, H.-L., Eur. J. Med.
Chem., 2015, vol. 94, pp. 447–457.
REFERENCES
1. Fustero, S., Sánchez-Roselló, M., Barrio, P., and
Simón-Fuentes, A., Chem. Rev., 2011, vol. 111,
pp. 6984–7034.
22. Mishra, C.B., Kumari, S., and Tiwari, M., Eur. J. Med.
https://doi.org/10.1021/cr2000459
Chem., 2015, vol. 92, pp. 1–34.
2. Ansari, A., Ali, A. and Asif, M., New J. Chem., 2017,
23. Ayati, A., Emami, S., Asadipour, A., Shafiee, A., and
Foroumadi, A., Eur. J. Med. Chem., 2015, vol. 97,
pp. 699–718.
vol. 41, pp. 16–41.
https://doi.org/10.1039/C6NJ03181A
3. Kumar, S., Bawa, S., Drabu, S., Kumar, R., and Gup-
ta, H., Recent Pat. Anti-Infect. Drug Discov., 2009,
vol. 4, pp. 154–163.
24. Sun, Z.Q., Tu, L.X., Zhuo, F.J., and Liu, S.X., Bioorg.
Med. Chem. Lett., 2016, vol. 26, pp. 747–750.
25. D’Ascenzio, M., Chimenti, P., Gidaro, M.C., De
Monte, C., De Vita, D., Granese, A., Scipione, L., Di
Santo, R., Costa, G., Alcaro, S., Yáñez, M., and Car-
radori, S., J. Enzyme Inhib. Med. Chem., 2015, vol. 30,
pp. 908–919.
4. Shaaban, M.R., Mayhoub, A.S., and Farag, A.M., Ex-
pert Opin. Ther. Pat., 2012, vol. 22, pp. 253–291.
5. Marella, A., Ali, R., Alam, T., Saha, R., Tanwar, O.,
Akhter, M., Shaquiquzzaman, M., and Alam, M.M.,
Mini-Rev. Med. Chem., 2013, vol. 13, pp. 921–931.
26. Zala, A.V., Walker, M.M., and Talley N.J., Expert
6. Alex, J.M. and Kumar, R., J. Enzyme Inhib. Med.
Opin. Emerg. Drugs, 2015, vol. 20, pp. 221–233.
Chem., 2014, vol. 29, pp. 427–442.
27. Logu, A.D., Sadd, M.I., Cardia, M.C., Borgna, R.,
Sanna, C., Saddi, B., and Elias, M., J. Antimi-
crob.Chemother., 2005, vol. 55, pp. 692–698.
7. Havrylyuk, D., Zimenkovsky, B., Vasylenko, O.,
Zaprutko, L., Gzella, A., and Lesyk, R., Eur. J. Med.
Chem., 2009, vol. 44, pp. 1396–1404.
8. Shaharyar, M., Abdullah, M.M., Bakht, M.A., and
Majeed, J., Eur. J. Med. Chem., 2010, vol. 45, pp. 114–
119.
9. Congiu, C., Onnis, V., Vesci, L., Castorina, M., and
Pisano, C., Bioorg. Med. Chem., 2010, vol. 18,
pp. 6238–6248.
28. Liu, C.L., Li, Z.M., and Zhong, B., J. Fluorine Chem.,
2004, vol. 125, pp. 1287–1290.
29. Kumar, R.V.K. and Kumar, V.S.R.S., J. Heterocycl.
Chem., 2005, vol. 42, pp. 1191–1193.
30. D’Andrea, S., Zheng, Z.B., DenBleyker, K., Fung-
Tomc, J.C., Yang, H., Clark, J., Taylor, D., and Bron-
son, J., Bioorg. Med. Chem. Lett., 2005, vol. 15,
pp. 2834–2839.
10. Al-Abdullah, E.S., Molecules, 2011, vol. 16, p. 3410–
3419.
31. Azarifar, D. and Shaebanzadeh, M., Molecules, 2002,
11. Bano, S., Javed, K., Ahmad, S., Rathish, I.G.,
Singh, S., and Alam, M.S., Eur. J. Med. Chem., 2011,
vol. 46, pp. 5763–5768.
12. Bashir, R., Ovais, S., Yaseen, S., Hamid, H., Alam, M.S.,
Samim, M., Singh, S., and Javed, K., Bioorg. Med.
Chem. Lett., 2011, vol. 21, pp. 4301–4305.
vol. 7, pp. 885–895.
32. Matysiak, J. and Niewiadom A.Y., Bioorg. Med. Chem.,
2003, vol. 11, pp. 2285–2287.
33. Jungheim, L.N., Sigmund, S.K., and Fisher, J.W., Tet-
rahedron Lett., 1987, vol. 28, pp. 285–288.
13. Amin, K.M., Eissa, A.A.M., Abou-Seri, S.M., 34. Temel, H.E., Altintop, M.D., and Özdemir, A., Turk.
Awadallah, F.M., and Hassan, G.S., Eur. J. Med.
Chem., 2013, vol. 60, pp. 187–198.
J. Pharm. Sci., 2018, vol. 15, pp. 333–338.
https://doi.org/10.4274/tjps.20982
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 46 No. 3 2020

392
SAFAA I. ELEWA et al.
35. Nassar, I.F., Atta-Allah, S.R., and Elgazwy, A-S.S.H.,
J. Enz. Inhib. Med. Chem., 2015, vol. 30, pp. 396–405.
https://doi.org/10.3109/14756366.2014.940936
pp. 1086–1100.
https://doi.org/10.1002/jhet.3496
40. Kabli, R.A., Khalaf, A.A., Zimaity, M.T., Khalil, A.M.,
Kaddah, A.M., and Al-Rifaie, H.A., J. Indian Chem.
Soc., 1991, vol. 68, pp. 47–51.
41. Wiley, R.H., Jarboe, C.H., Hayes, F.N., Hansbury, E.,
Nielsen, J.T., and Callahan, P.X., J. Org. Chem., 1958,
Vol. 23, pp. 732–738.
36. Nassar, I.F., El Farargy, A.F., Abdelrazek, F.M., and
Ismail, N.S.M., Nucleosides, Nucleotides Nucleic Acids,
2017, vol. 36, pp. 275–291.
https://doi.org/10.1080/15257770.2016.1276290
37. Nassar, I.F., El Farargy, A.F., and Abdelrazek, F.M., J.
Heterocycl. Chem., 2018, vol. 55, pp. 1709–1718.
https://doi.org/10.1002/jhet.3208
42. Abdel-Kader, H.A. and Seddkey, S.R., Assiut Vet. Med.
J., 1995, vol. 34, pp. 67.
43. Fagbemi, J., Ferdinand, L., and Adenipekun, T., Afr. J.
Biotech., 2009, vol. 8, pp. 1176–1182.
38. Nassar, I.F., Atta-Allah, S.R., and Hemdan M.M.,
Phosphorus Sulphur, 2018, vol. 193, pp. 630–636.
https://doi.org/10.1080/10426507.2018.1487435
44. Skehan, P. and Storeng, R., J. Natl. Cancer Inst., 1990,
vol. 82, pp. 1107–1112.
39. Nassar, I.F., EL-Kady D.S., Awad H.M., and El- 45. Eweiss, N.F. and Osman, A., J. Heterocycl. Chem.,
Sayed W.A., J. Heterocycl. Chem., 2019, vol. 56, 1980, vol. 17, pp. 1713–1717.
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY
Vol. 46
No. 3
2020