Design, Synthesis, and In Vitro Cytotoxic Activity of Certain 2-[3-Phenyl-4-(pyrimidin-4-yl)-1H-pyrazol1-yl]acetamide Derivatives

Article abstract of DOI:10.1134/S1070428020030239

Abstract: With a view to finding new anticancer agents, different aryloxy groups wereattached to C² of the pyrimidine ring in2-[3-phenyl-4-(pyrimidin-4-yl)-1H-pyrazol-1-yl]acetamide in five steps using methyl3-methoxy-5-methylbenzoate as the ke

Full text of DOI:10.1134/S1070428020030239

ISSN 1070-4280, Russian Journal of Organic Chemistry, 2020, Vol. 56, No. 3, pp. 514–520. © Pleiades Publishing, Ltd., 2020.
Design, Synthesis, and In Vitro Cytotoxic Activity
of Certain 2-[3-Phenyl-4-(pyrimidin-4-yl)-1H-pyrazol-
1-yl]acetamide Derivatives
M. M. Al-Saneaa, D. G. T. Parambia, M. E. Shakerb,c, H. A. M. Elsherifd, H. A. H. Elshemye,
R. B. Bakra,e, T. I. M. Al-Warhif, M. Gamala,g, and M. A. Abdelgawada,e,*
a Department of Pharmaceutical Chemistry, College of Pharmacy, Jouf University, Sakaka, Aljouf, 2014 Saudi Arabia
b Department of Pharmacology, College of Pharmacy, Jouf University, Sakaka, Aljouf, 2014 Saudi Arabia
c Department of Pharmacology and Toxicology, Faculty of Pharmacy, Mansoura University, Mansoura, 35516 Egypt
d Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Deraya University, Minya, 51619 Egypt
e Department of Pharmaceutical Organic Chemistry, Beni-Suef University, Beni-Suef, 62514 Egypt
f Department of Chemistry, College of Science, Princess Nourah bint Abdulrahman University,
Riyadh, 11671 Saudi Arabia
g Department of Pharmaceutical Analytical Chemistry, Beni-Suef University, Beni-Suef, 62514 Egypt
*e-mail: mohamedabdelwahab976@yahoo.com; mhmdgwd@ju.edu.sa
Received November 11, 2019; revised December 18, 2019; accepted December 28, 2019
Abstract—With a view to finding new anticancer agents, different aryloxy groups were attached to C2 of the
pyrimidine ring in 2-[3-phenyl-4-(pyrimidin-4-yl)-1H-pyrazol-1-yl]acetamide in five steps using methyl
3-methoxy-5-methylbenzoate as the key intermediate product. The anticancer activity of the synthesized
compounds was tested on 60 cancer cell lines at 10 μM. One compound showed an appreciable cancer cell
growth inhibition (about 20%) against eight cancer cell lines (HOP-92, NCI-H226, 79SNB-75, A498, SN12C,
UO-31, T-47D, and MDA-MB-468).
Keywords: 3-phenyl-4-pyrimidinyl-1H-pyrazole, acetamide analogs, anticancer activity
DOI: 10.1134/S1070428020030239
Cancer is a genetic disease acquired by the accu-
mulation of mutated genes responsible for cell prolif-
eration and survival. The incidence of new types of
human neoplasms has alarmingly increased, resulting
in a big public health crisis and the leading cause of
death. Because of a series of genetic defects, cancer
leads to the transformation of normal cells into
cancerous ones associated with abnormalities in the
regulation of cell proliferation, growth, and survival
processes [1]. Thus, there is a critical need to develop
new and innovative potent anticancer agents. Different
kinds of classical cancer therapies, such as chemo-
therapy, irradiation, and immunotherapy have achieved
significant improvements in the prevention and control
of different human malignancies; however, the emer-
gence of severe toxicity has restricted their application.
Due to the reported toxicity of classical anticancer
drugs, many selective less toxic anticancer drugs have
been extensively studied [2–5].
Phenylpyridinyl and phenylpyrimidinyl pyrazole
scaffold-based compounds are promising anti-prolif-
erative agents targeting different types of human
cancers [5–12]. The purpose of this study was to dis-
cover newer and efficient anti-proliferative agents for
further developments [12–15]. Herein, we report the
synthesis of a series of 2-[3-phenyl-4-(pyrimidin-4-yl-
1H-pyrazol-1-yl]acetamide derivatives with different
aryloxy substituents at the 2-position of the pyrimidine
ring, which were designed in coherence with the lead
compound A [14] (Scheme 1). The synthesized com-
pounds were subjected to primary screening over
a panel of 60 cancer cell lines at a single dose con-
centration of 10 μM.
The target diarylpyrazole-based compounds were
synthesized through methyl 3-methoxy-5-methylben-
zoate (4) as the key intermediate product, which
was prepared according to the reported procedures
[12, 15–19] (Scheme 2). The first step was condensa-
514

DESIGN, SYNTHESIS, AND IN VITRO CYTOTOXIC ACTIVITY
515
Scheme 1.
OH
OMe
O
Me
N
Me
N
NH2
N
N
CN
N
N
N
N
N
OAr
Lead compound A
Designed compounds
tion of diethyl oxalate with acetone in the presence of
NaOEt in anhydrous ethanol, which produced ethyl
2-hydroxy-4-oxopent-2-enoate sodium salt (1) [20].
their antineoplastic activities. The compounds were
subjected to in vitro anticancer assay against a panel of
60 different human tumor cell lines representing nine
different tissues (leukemia, melanoma, and lung, colon,
brain, ovarian, breast, prostate, and renal cancers). The
compounds were tested at a single dose concentration
of 10 μM, and the cultures were incubated for 48 h.
The results for each compound were represented as
mean graphs of the percent growth of the treated cells
relative to the untreated cells (control). The percent cell
growth of 19 most sensitive cell lines treated with
compounds 9a–9h at a concentration of 10 мM is
illustrated by Table 1.
Salt 1 was then heated in acetic acid, and subsequent
acidification with sulfuric acid gave tetrahydrofuran-
2,3-dione derivative 2. Compound 2 underwent rear-
rangement and aromatization into 3-hydroxy-5-methyl-
benzoic acid (3) on heating with magnesium oxide in
boiling water for 45 min, followed by acidification
with hydrochloric acid [21]. Acid 3 was treated with
methyl iodide in boiling acetone in the presence of
excess potassium carbonate and a catalytic amount of
DMAP to obtain methyl 3-methoxy-5-methylbenzoate
(4) in 94% yield (cf. [22]). Ester 4 was reacted with
lithium salt derived from 4-methyl-2-(methylsulfanyl)-
pyrimidine by the action of lithium bis(trimethylsilyl)-
amide (LiHMDS) in anhydrous THF at room tempera-
ture. The resulting tautomeric α,β-unsaturated ketone 5
was converted to pyrazole derivative 6 by heating with
excess N,N-dimethylformamide dimethyl acetal for
12 h, followed by cyclization with hydrazine hydrate in
anhydrous ethanol. The reaction of 6 with iodoacet-
amide in the presence of excess potassium carbonate
produced pyrazolylacetamide 7 as a single isomer.
2-{4-[2-(Methanesulfonyl)pyrimidin-4-yl]-3-(3-me-
thoxy-5-methylphenyl)-1H-pyrazol-1-yl}acetamide (8)
was prepared in a good yield (70%) by oxidation of 7
with m-chloroperoxybenzoic acid and magnesium sul-
fate in anhydrous methylene chloride for 2 h. Finally,
compound 8 was subjected to nucleophilic substitution
reaction with different substituted aromatic hydroxy
compounds on heating at 80°C in anhydrous dimethyl-
formamide for 3 h.
All the compounds tested displayed a growth inhib-
itory profile of <27% in 19 most sensitive tumor cell
lines. Compound 9h showed 25.4 and 20.8% growth
inhibition toward SNB-75 and A498 cell lines, respec-
tively. Compound 9a with a trimethoxyphenyl moiety
exhibited 26% growth inhibition against breast cancer
cell line MDA-MB-468. Compound 9b exhibited 21%
inhibition on NCI-H226 and MDA-MB-468 cell lines
and 22.2% growth inhibition on SNB-75. Compound
9c restricted the growth of renal A498 cells by 26.7%.
This was the maximum inhibitory effect observed
among the synthesized molecules against the tested cell
lines. It also showed 17.3% growth inhibition in breast
cancer cell line MDA-MB-468, and compound 9f
prevented the growth of NSCLC-HOP-92 and UO-31
cells by 25.9 and 20.3%, respectively. On the other
hand, compounds 9d and 9e did not exert any appre-
ciable effect. Similarly, compound 9g specifically
retarded the growth of renal cancer cell lines A498 and
UO-31 by 23.7 and 20.2%, respectively.
The structures of 9a–9h were selected by the
National Cancer Institute (NCI, Bethesda, Maryland,
USA) on the basis of the degree of structural variation
and computer modeling techniques for evaluation of
Based on the threshold inhibition criteria, com-
pounds 9a–9h displayed no any significant cytotoxic
activity. However, 3,4-dimethoxyphenyl derivative 9b
was found to be effective as it showed up to 22.2%
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AL-SANEA et al.
516
Scheme 2.
O
O
H
Me
O
Me
O
O
O
O
O
O
i
ii
OEt
Me
+
EtO
EtO
O
O
EtO
EtO
Me
Me
O
O
O
ONa
O
O
O
Me
Me
1
2
OMe
OMe
SMe
N
N
, v
COOH
COOMe
O
OH
Me
Me
iii
iv
Me
HO
Me
MeO
Me
N
N
N
N
SMe
SMe
3
4
5
OMe
OMe
OMe
N
N
N
Me
Me
Me
vi
vii
viii
NH
N
N
NH2
NH2
O
O
N
N
N
N
N
N
SMe
SMe
SO2Me
6
7
8
OMe
3'
4'
5'
2'
1'
3
N
Me
ix
6'
N
4
5"
6"
NH2
5
4"
O
N
N
2"
OAr
9a–9h
9, Ar = 3,4,5-(MeO)3C6H2 (a), 3,4-(MeO)2C6H3 (b), pyridin-3-yl (c), pyridin-4-yl (d), 1-oxidopyridin-1-ium-4-yl (e), 4-MeOC6H4 (f),
2-EtOC6H4 (g), pyridin-2-yl (h). Reagents and conditions: i: NaOEt, EtOH, r.t.,4 h, yield 87%; ii: AcOH–H2O (1:1), r.t., 2 h, 50%;
iii: MgO, H2O, reflux, 45 min, 42%; iv: K2CO3, MeI, DMAP, acetone, 65°C, 12 h, 94%; v: LiHMDS, THF, N2, r.t., 24 h; vi:
(1) DMFDMA, 90°C, 20 h; (2) N2H4·H2O, EtOH, r.t., 12 h, 79%; vii: K2CO3, ICH2CONH2, DMF, reflux, 2 h, 43%; viii: m-CPBA,
MgSO4, CH2Cl2, r.t., 2 h ; ix: ArOH, DMF, 80°C, 3 h.
growth inhibition in 19 sensitive cell lines. The reduced
cytotoxic activity may be due to hindrances in the
structural orientation to adopt a conformation/pose
favoring the required activity profile. Taking compound
9b as a template, further modifications are in progress
to achieve a better antiproliferative activity.
In summary, we have designed, synthesized, and
characterized a series of 2-[3-phenyl-4-(pyrimidin-4-
yl)-1H-pyrazole]acetamide derivatives, and their in
vitro antiproliferative activity has been assessed against
60 NCI cell lines at a single dose concentration of
10 μM. One of the synthesized compounds, namely
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DESIGN, SYNTHESIS, AND IN VITRO CYTOTOXIC ACTIVITY
517
Table 1. Percent growth of cells treated with compounds 9a–9h (10 мM) for 19 most sensitive tumor cell linesa
Cancer cell line
K-562
9a
9b
9c
9d
9e
9f
9g
9h
NI
NI
89
87
95
97
88
98
NI
97
99
93
NI
95
92
96
NI
NI
74
NI
88
78
79
87
88
78
99
92
95
87
83
89
82
88
93
NI
80
80
NI
NI
82
97
99
96
93
96
93
99
NI
73
99
96
93
89
NI
NI
83
NI
NI
95
99
92
98
98
99
96
95
NI
96
97
NI
90
NI
NI
94
NI
NI
99
96
NI
NI
NI
96
NI
95
NI
NI
92
NI
92
95
98
89
NI
NI
88
NI
74
NI
92
94
94
89
88
NI
NI
98
95
93
80
91
NI
92
92
NI
93
78
93
88
90
83
NI
87
89
93
76
87
94
80
94
NI
83
97
NI
NI
89
NI
NI
93
74
93
98
90
NI
79
94
NI
92
96
97
98
93
Leukemia
NSCLC
SR
HOP-92
NCI-H226
SF-295
CNS cancer
SF-539
SNB-75
LOX IMVI
UACC-62
OVCAR-4
OVCAR-5
A498
CAKI-1
SN12C
UO-31
Melanoma
Ovarian cancer
Renal cancer
Prostate cancer
Breast cancer
PC-3
HS 578T
T-47D
MDA-MB-468
a
NI stands for no inhibition or increase of cell growth.
2-{4-[2-(3,4-dimethoxyphenoxy)pyrimidin-4-yl]-3-(3-
methoxy-5-methylphenyl)-1H-pyrazol-1-yl}acetamide
showed an appreciable cytotoxicity (up to 22.2% cell
growth inhibition) against eight cancer cell lines
covering four human malignancies (NSCLC, CNS,
renal, and breast cancers). It could be considered as
a promising hit compound and is currently subjected to
further minor structural modifications to improve its
antiproliferative potency.
added to a solution of 4-[3-(3-methoxy-5-methyl-
phenyl)-1H-pyrazol-4-yl]-2-(methylsulfanyl)pyrimi-
dine (6, 5 g, 16 mmol) in DMF (65 mL). The mixture
was heated at 50°C on an oil bath for 16 h, cooled, and
treated with brine and ethyl acetate. The organic layer
was separated, and the aqueous phase was extracted
with ethyl acetate. The combined organic layers were
washed with brine, dried over MgSO4, and concen-
trated under reduced pressure. The residue was purified
by column chromatography using hexane–ethyl acetate
EXPERIMENTAL
1
(4:1). Yield 43%, mp 265–267°C. H NMR spectrum
(DMSO-d6), δ, ppm: 2.31 s (3H, CH3), 2.37 s (3H,
SCH3), 3.73 s (3H, OCH3), 4.88 s (2H, CH2CO), 6.81–
6.83 m (2H, 2′-H, 4′-H), 6.90 s (1H, 6′-H), 6.98 d (J =
5.6 Hz, 1H, 5″-H), 7.36 s (1H, 5-H), 8.44 s (2H, NH2),
8.45 d (1H, 6″-H, J = 5.6 Hz). 13C NMR spectrum
(DEPT135, DMSO-d6), δC, ppm: 13.58 (SCH3),
21.52 (CH3), 54.46 (OCH3), 55.51 (CH2), 111.71 (C4),
113.42 (C2′), 115.11 (C4′), 121.15 (C6′), 122.4 (C5″),
131.76 (C5), 135.36 (C1′), 139.51 (C5′), 155.98 (C3),
157.79 (C6″), 162.43 (C3′), 164.54 (C4″), 168.24
(C2″), 173.52 (C=O). Found, %: C 58.70; H 5.10;
N 18.90. C18H19N5O2S. Calculated, %: C 58.52;
H 5.18; N 18.96.
1
The H and 13C NMR spectra were recorded on
a Bruker Avance 400 spectrometer at 400 and
100 MHz, respectively. The melting points were meas-
ured with a Thomas Hoover capillary melting apparatus
and are uncorrected. Column chromatography was
performed on Merck silica gel 60 (230–400 mesh).
Thin-layer chromatography was carried out using glass
sheets pre-coated with silica gel 60 F254 (Merck). All
commercial reagents were obtained from Aldrich and
Tokyo Kasei Chemicals and generally used without
further purification. Compounds 1–6 were prepared
according to the reported methods [16–18].
2-{3-(3-Methoxy-5-methylphenyl)-4-[2-(methyl-
sulfanyl)pyrimidin-4-yl]-1H-pyrazol-1-yl}acetamide
(7). Iodoacetamide (2.96 g, 40 mmol) and freshly
ground potassium carbonate (5.52 g, 40 mmol) were
2-{4-[2-(Methanesulfonyl)pyrimidin-4-yl]-3-(3-
methoxy-5-methylphenyl)-1H-pyrazol-1-yl}acet-
amide (8). A mixture of m-chloroperoxybenzoic acid
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AL-SANEA et al.
518
(4.71 g, 27.3 mmol) and MgSO4 (6.57 g, 54.6 mmol) in
methylene chloride (100 mL) was stirred for 1 h at
room temperature. 2-{3-(3-Methoxy-5-methylphenyl)-
4-[2-(methylsulfanyl)pyrimidin-4-yl]-1H-pyrazol-1-
yl}acetamide (7, 1.6 g, 4.55 mmol) was then added,
and the mixture was stirred at room temperature for
2 h. The mixture was filtered, the precipitate was
washed with methylene chloride, the filtrate was com-
bined with the washings, washed with water and brine,
dried over MgSO4, and concentrated under reduced
pressure. The residue was purified by column chroma-
tography using hexane–ethyl acetate (1:1) as eluent.
Yield 70%, white solid, mp 280–282°C. 1H NMR spec-
trum (CDCl3), δ, ppm: 2.39 s (3H, CH3), 3.31 s (3H,
SCH3), 3.83 s (3H, OCH3), 4.92 s (2H, CH2CO), 6.86 s
(1H, 2′-H), 6.93 s (1H, 4′-H), 6.99 s (1H, 2′-H), 7.31 d
(1H, 5″-H, J = 5.6 Hz), 8.39 s (1H, 5-H), 8.56 s (2H,
NH2), 8.63 d (1H, 6″-H, J = 5.6 Hz). 13C NMR
spectrum (DEPT135, CDCl3), δC, ppm: 21.53 (CH3),
38.95 (SO2CH3), 55.39 (OCH3), 55.50 (CH2), 111.42
(C4), 115.85 (C2′), 119.56 (C4′), 122.02 (C6′), 132.32
(C5″), 131.52 (C5), 134.88 (C1′), 140.53 (C5′), 154.32
(C3), 157.96 (C6″), 162.27 (C3′), 164.44 (C4″), 168.05
(C2″), 173.11 (C=O). Found, %: C 53.80; H 4.90;
N 17.50. C18H19N5O2S. Calculated, %: C 53.85;
H 4.77; N 17.45.
161. 83 (C2″), 168.26 (C=O). Found, %: C 61.70;
H 5.5; N 14.00. C26H27N5O6. Calculated, %: C 61.77;
H 5.38; N 13.85.
2-{4-[2-(3,4-Dimethoxyphenoxy)pyrimidin-4-yl]-
3-(3-methoxy-5-methylphenyl)-1H-pyrazol-1-yl}-
acetamide (9b). Yield 2.68 g (70%), mp ˃300°C.
1H NMR spectrum (CDCl3), δ, ppm: 2.38 s (3H, CH3),
3.80 s (3H, OCH3), 3.81 s (3H, OCH3), 3.92 s (3H,
OCH3), 4.85 s (2H, CH2CO), 6.82–6.86 m (5H, 2′-H,
2′′′-H, 4′-H, 5′′′-H, 6′′′-H), 6.90 d (1H, 5″-H, (J =
5.2 Hz), 6.99 s (2H, NH2), 7.09 s (1H, 6′-H), 8.16 s
(1H, 5-H), 8.29 d (J = 5.2 Hz, 1H, 6″-H). 13C NMR
spectrum (CDCl3), δC, ppm: 21.48 (CH3), 55.02
(OCH3), 55.32 (CH2), 56.07 (OCH3), 56.21 (OCH3),
122.34 (C4), 109.73 (C2′′′), 111.47 (C2′), 112.75 (C4′),
115.21 (C5″), 115.71 (C6′′′), 119.36 (C6′), 122.15 (C4′′′),
123.61 (C5′′′), 133.38 (C5), 134.27 (C1′), 140.04 (C5′),
146.52 (C3′′′), 147.43 (C1′′′), 152.68 (C3), 153.87 (C6″),
159.12 (C3′), 159.77 (C4″), 161.60 (C2″), 168.59 (C=O).
Found, %: C 63.10; H 5.4; N 14.70. C25H25N5O6.
Calculated, %: C 63.15; H 5.30; N 14.73.
2-{3-(3-Methoxy-5-methylphenyl)-4-[2-(pyridin-
3-yloxy)pyrimidin-4-yl]-1H-pyrazol-1-yl}acetamide
1
(9c). Yield 2.11 g (65%), mp ˃300°C. H NMR
spectrum (CDCl3), δ, ppm: 2.39 s (3H, CH3), 3.82 s
(3H, OCH3), 4.88 s (2H, CH2CO), 6.85 s (2H, NH2),
6.92 s (1H, 2′-H),6.93 s (1H, 4′-H), 7.38–7.42 m (1H,
5′′′-H), 7.59–7.62 m (2H, 4′′′-H, 6′′′-H), 8.13 s (1H,
6′-H), 8.34 d (1H, 5″-H, J = 5.6 Hz), 8.16 s (1H, 5-H),
8.54 d (1H, 6″-H, J = 5.6 Hz), 8.62 s (1H, 2′′′-H).
13C NMR spectrum (DEPT135, CDCl3), δC, ppm:
21.53 (CH3), 55.06 (OCH3), 55.35 (CH2), 99.69 (C4),
111.54 (C2′), 113.34 (C4′), 115.68 (C5″), 119.34 (C6′),
122.10 (C5′′′), 129.26 (C5), 133.26 (C3′′′), 133.85 (C4′′′),
134.11 (C1′), 139.62 (C5′), 143.90 (C6′′′), 146.21 (C2′′′),
152.23 (C3), 153.12 (C6″), 159.58 (C3′), 159.41 (C4″),
161.15 (C2″), 168.29 (C=O). Found, %: C 63.50;
H 4.80; N 20.20. C22H20N6O3. Calculated, %: C 63.45;
H 4.84; N 20.18.
General procedure for the synthesis of
2-[4-(2-R-oxypyrimidin-4-yl)-3-(3-methoxy-5-
methylphenyl)-1H-pyrazol-1-yl]acetamides 9a–9h.
A mixture of compound 8 (7.8 mmol) and aromatic
hydroxy compound (7.8 mmol) in anhydrous DMF
(5 mL) was heated under reflux at 80°C in an oil bath
for 3 h. The mixture was cooled, diluted with ethyl
acetate, and washed with saturated aqueous ammonium
chloride and brine. The organic layer was dried over
MgSO4 and concentrated under reduced pressure, and
the residue was purified by column chromatography.
2-{3-(3-Methoxy-5-methylphenyl)-4-[2-(3,4,5-tri-
methoxyphenoxy)pyrimidin-4-yl]-1H-pyrazol-1-yl}-
acetamide (9a). Yield 2.36 g (60%), mp ˃300°C.
1H NMR spectrum (CDCl3), δ, ppm: 2.38 s (3H, CH3),
3.79 s (3H, OCH3), 3.82 s (6H, OCH3), 3.83 s (3H,
OCH3), 4.87 s (2H, CH2CO), 6.13 s (2H, C6H2), 6.49–
6.51 m (2H, 2′-H, 4′-H), 6.87 d (1H, 5″-H, J = 5.6 Hz),
6.88 s (2H, NH2), 6.96 s (1H, 6′-H), 8.19 s (1H, 5-H),
8.31 d (1H, 6″-H, J = 5.2 Hz). 13C NMR spectrum
(CDCl3), δC, ppm: 21.39 (CH3), 55.03 (OCH3), 55.30
(CH2), 56.06 (2C, OCH3), 56.22 (OCH3), 93.51 (C2′′′,
C6′′′), 99.64 (C4), 111.79 (C2′), 112.86 (C4′), 115.62
(C5″), 119.36 (C6′), 122.18 (C4′′′), 133.43 (C5), 134.06
(C1′), 139.99 (C5′), 147.22 (C1′′′), 149.09 (C3′′′, C5′′′),
152.65 (C3), 153.67 (C6″), 159.21 (C3′), 159.85 (C4″),
2-{3-(3-Methoxy-5-methylphenyl)-4-[2-(pyridin-
4-yloxy)pyrimidin-4-yl]-1H-pyrazol-1-yl}acetamide
1
(9d). Yield 1.78 g (55%), mp ˃300°C. H NMR spec-
trum (CDCl3), δ, ppm: 2.39 s (3H, CH3), 3.82 s (3H,
OCH3), 4.94 s (2H, CH2CO), 6.44 d (2H, 3′′′-H, 5′′′-H,
J = 8 Hz), 6.88 s (2H, NH2), 6.95 s (1H, 2′-H), 7.09 s
(1H, 4′-H), 7.13 d (1H, 5″-H, J = 5.2 Hz), 8.19 s (1H,
6′-H), 8.28 s (1H, 5-H), 8.51 d (1H, 6″-H, J = 5.6 Hz),
8.66 d (2H, 2′′′-H, 6′′′-6, J = 8 Hz). 13C NMR spectrum
(DEPT135, CDCl3), δC, ppm: 21.53 (CH3), 55.16
(OCH3), 55.34 (CH2), 112.27 (C2′), 114.64 (C4′),
115.22 (C5″), 111.35 (C4), 119.67 (C6′), 122.25 (C3′′′,
C5′′′), 129.77 (C5), 133.57 (C2′′′, C6′′′), 134.72 (C1′),
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DESIGN, SYNTHESIS, AND IN VITRO CYTOTOXIC ACTIVITY
519
139.53 (C5′), 143.90 (C4′′′), 152.75 (C3), 153.46 (C6″),
159.43 (C3′), 159.41 (C4″), 161.15 (C2″), 168.29 (C=O).
Found, %: C 63.40; H 4.90; N 20.10. C22H20N6O3.
Calculated, %: C 63.45; H 4.84; N 20.18.
21.23 (CH3), 55.17 (OCH3), 60.64 (CH2), 65.11 (CH2),
104.23 (C4), 111.23 (C2′), 112.78 (C4′), 115.23 (C5″),
115.88 (C3′′′), 120.45 (C6′), 122.65 (C6′′′), 123.45 (C2′′′),
125.55 (C4′′′), 131.34 (C3′′′), 133.55 (C5), 134.19 (C1′),
140.24 (C5′), 146.32 (C1′′′), 152.46 (C3), 153.34 (C6″),
159.69 (C3′), 159.21 (C4″), 161.08 (C2″), 168.99 (C=O).
Found, %: C 65.60; H 5.20; N 15.30. C25H25N5O4.
Calculated, %: C 65.35; H 5.48; N 15.24.
3-({[4-(1-(2-Amino-2-oxoethyl)-3-(3-methoxy-5-
methylphenyl)-1H-pyrazol-4-yl]pyrimidin-2-yl}-
oxy)pyridine 1-oxide (9e). Yield 2.01 g (60%),
1
mp ˃300°C. H NMR spectrum (DMSO-d6), δ, ppm:
2-{3-(3-Methoxy-5-methylphenyl)-4-[2-(pyridin-
2.30 s (3H, CH3), 3.74 s (3H, OCH3), 4.88 s (2H,
CH2CO), 6.79 s (2H, NH2), 6.85 s (1H, 2′-H), 7.07 d
(1H, 5″-H, J = 5.2 Hz), 7.25 d (1H, 4′′′-H, J = 8.8 Hz),
7. 37 s (1H, 2′′′-H), 7.39 d (1H, 6′′′-H, J = 6.8 Hz),
7.67 s (1H, 4′-H), 8.33 s (1H, 6′-H), 8.34 t (1H, 5′′′-H,
J = 2 Hz), 8.37 s (1H, 5-H), 8.59 d (1H, 6″-H, J =
5.2 Hz). 13C NMR spectrum (APT, DMSO-d6), δC,
ppm: 21.51 (CH3), 54.45 (OCH3), 55.52 (CH2), 99.84
(C4), 111.76 (C2′), 114.03 (C4′), 115.16 (C5″), 117.41
(C4′′′), 119.93 (C6′), 122.27 (C5′′′), 126.47 (C5), 134.13
(C2′′′), 134.42 (C1′), 135.48 (C5′), 136.43 (C6′′′), 139.41
(C3′′′), 150.59 (C3), 151.75 (C6″), 159.46 (C3′), 160.15
(C4″), 162.54 (C2″), 168.47 (C=O). Found, %: C 61.10;
H 4.60; N 19.40. C22H20N6O4. Calculated, %: C 61.10;
H 4.66; N 19.43.
2-yloxy)pyrimidin-4-yl]-1H-pyrazol-1-yl}acetamide
1
(9h). Yield 1.62 g (50%), mp ˃300°C. H NMR spec-
trum (CDCl3), δ, ppm: 2.39 s (3H, CH3), 3.82 s (3H,
OCH3), 4.88 s (2H, CH2CO), 5.91 s (1H, 5′′′-H), 6.54 s
(1H, 2′-H), 6.68 d (1H, 6′′′-H, J = 8.8 Hz), 6.85 s (1H,
4′-H), 6.88 s (2H, NH2), 6.96 s (1H, 6′-H), 7.18 d (1H,
5″-H, J = 5.2 Hz), 7.41–7.43 m (1H, 4′′′-H), 7.61 d
(1H, 3′′′-H, J = 6.4 Hz), 8.31 s (1H, 5-H), 8.63 d (1H,
6″-H, J = 5.6 Hz). 13C NMR spectrum (APT,
DMSO-d6), δC, ppm: 21.52 (CH3), 55.00 (OCH3),
55.38 (CH2), 105.79 (C4), 111.52 (C2′), 115.73 (C4′),
116.53 (C5″), 118.77 (C6′), 122.14 (C4′′′), 122.63 (C5′′′),
133.24 (C1′), 134.55 (C5′), 135.92 (C5), 140.25 (C2′′′),
140.37 (C3), 152.65 (C6″), 158.77 (C3′′′), 159.33 (C3′),
159.87 (C4″), 161.32 (C1′′′), 162.00 (C2″), 168.37
(C=O). Found, %: C 63.60; H 4.70; N 20.30.
C22H20N6O3. Calculated, %: C 63.45; H 4.84; N 20.18.
2-{3-(3-Methoxy-5-methylphenyl)-4-[2-(4-
methoxyphenoxy)pyrimidin-4-yl]-1H-pyrazol-1-yl}-
acetamide (9f). Yield 2.25 g (65%), mp ˃300°C.
1H NMR spectrum (CDCl3), δ, ppm: 2.39 s (3H, CH3),
3.83 s (3H, OCH3), 3.87 s (3H, OCH3), 4.89 s (2H,
CH2CO), 6.11 d (2H, 2′′′-H, 6′′′-H, J = 7.2 Hz), 6.39 d
(2H, 3′′′-H, 5′′′-H, J = 7.2 Hz), 6.86 s (2H, NH2), 6.91 s
(1H, 2′-H), 6.95-6.98 m (2H, 4′-H, 6′-H), 7.98–8.17 m
(2H, 5″-H, 5-H), 8.17 d (1H, 6″-H, J = 5.6 Hz).
13C NMR spectrum (CDCl3), δC, ppm: 21.23 (CH3),
55.17 (OCH3), 56.03 (OCH3), 60.64 (CH2), 103.45
(C4), 111.67 (C2′), 112.65 (C4′), 115.73 (C5″), 115.88
(C3′′′, C5′′′), 119.19 (C6′), 123.61 (C2′′′, C6′′′), 133.73
(C5), 134.15 (C1′), 140.09 (C5′), 146.32 (C1′′′), 152.68
(C3), 153.65 (C6″), 154.87 (C4′′′), 159.12 (C3′), 159.74
(C4″), 161.45 (C2″), 168.77 (C=O). Found, %: C 64.60;
H 5.10; N 15.50. C24H23N5O4. Calculated, %: C 64.70;
H 5.20; N 15.72.
Cytotoxicity assay. The cell line screening was
performed at the National Cancer Institute (NCI,
Bethesda, Maryland, USA; www.dtp.nci.nih.gov) by
MTT assay according to the reported procedure [23].
FUNDING
This research was supported by the Jouf University
(grant no. 40/150) and by Korea Institute of Science and
Technology (2019 KIST School partnership project).
CONFLICT OF INTEREST
The authors declare the absence of conflict of interest.
REFERENCES
2-{4-[2-(2-Ethoxyphenoxy)pyrimidin-4-yl]-3-(3-
methoxy-5-methylphenyl)-1H-pyrazol-1-yl}acet-
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13C NMR spectrum (CDCl3), δC, ppm: 14.32 (CH3),
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