Synthesis and anticancer activity of benzotriazole derivatives

Article abstract of DOI:10.1002/jhet.3859

A series of benzotriazole (BTA) derivatives were synthesized as tyrosine protein kinase inhibitors using fragment-based design strategy. All desired compounds were synthesized with the reaction of benzotriazole, chloroacetonitrile and aromatic aldehyde using Ultrasonic-Microwave method and characterized by IR, ¹H and ¹³C-NMR, mass spectrometry (MS) and elemental analysis. The anticancer activity of these compounds was evaluated by CCK-8 method against carcinoma VX2, lung cancer A549, stomach cancer cell lines MKN45 and MGC in vitro. The results showed that all compounds showed good antiproliferative activity. In particular, compound 2.1 showed the most prominent inhibition of VX2 cell lines with IC₅₀ of 3.80 ± 0.75 μM. Compound 2.2 exhibited highly potent anticancer activity of stomach MGC cell lines with IC₅₀ of 3.72 ± 0.11 μM. A549 and MKN45 cell lines were sensitive to compound 2.5 with IC₅₀ of 5.47 ± 1.11 and 3.04 ± 0.02 μM, respectively.

Full text of DOI:10.1002/jhet.3859

Received: 10 September 2019
DOI: 10.1002/jhet.3859
Revised: 19 November 2019
Accepted: 3 December 2019
A R T I C L E
Synthesis and anticancer activity of benzotriazole
derivatives
Qiujing Li1
| Guijun Liu2,3 | Ningning Wang2,3 | Huiyong Yin2,3 | Zhulai Li4
1Department of Pharmacy, Zhangzhou
Health Vocational College, Zhangzhou,
China
2CAS Key Laboratory of Nutrition,
Metabolism and Food Safety, Shanghai
Institute of Nutrition and Health, Chinese
Academy of Science (CAS), Shanghai,
China
3Institute of Nutrition and Health,
University of Chinese Academy of
Sciences, Beijing, China
4College of Pharmacy, Fujian Medical
University, Fuzhou, China
Abstract
A series of benzotriazole (BTA) derivatives were synthesized as tyrosine pro-
tein kinase inhibitors using fragment-based design strategy. All desired com-
pounds were synthesized with the reaction of benzotriazole, chloroacetonitrile
and aromatic aldehyde using Ultrasonic-Microwave method and characterized
1
by IR, H and 13C-NMR, mass spectrometry (MS) and elemental analysis. The
anticancer activity of these compounds was evaluated by CCK-8 method
against carcinoma VX2, lung cancer A549, stomach cancer cell lines MKN45
and MGC in vitro. The results showed that all compounds showed good antip-
roliferative activity. In particular, compound 2.1 showed the most prominent
inhibition of VX2 cell lines with IC50 of 3.80 0.75 μM. Compound 2.2
exhibited highly potent anticancer activity of stomach MGC cell lines with
IC50 of 3.72 0.11 μM. A549 and MKN45 cell lines were sensitive to com-
pound 2.5 with IC50 of 5.47 1.11 and 3.04 0.02 μM, respectively.
Correspondence
Qiujing Li, Department of Pharmacy,
Zhangzhou Health Vocational College,
Zhangzhou 363000, China.
Email: qjlqj2009@163.com
1 | INTRODUCTION
binding modes, thereby specifically inhibiting its phos-
phorylation and inducing apoptosis of various cancer
The benzotriazole (BTA) family has emerged as a group
of pharmacophoric molecules with a broad spectrum of
cells.[14] Then Makowska et al synthesized more active
compounds in 2011 by structural modification of benzene
rings and introducing side chains connecting three car-
bons into 4-methyl-5,6,7-tribromo-1H-benzotriazole and
4-ethyl-5,6,7-tribromo-1H-benzotriazole.[15] In 2013, a
series of 1,3,4-oxadiazole derivatives docking with BTA
groups and their anticancer activity were reported. Some
of them have strong inhibitory activity against FAK,
which is comparable to cisplatin in the treatment of
human breast cancer.[6] A series of 4,5,6,7-tetra- bromo-
2-(3-chloropropyl)-2H-benzotriazole derivatives were syn-
thesized in 2016. The effect of these compounds on
human recombinant casein kinase 2α subunit (rhCK2α)
and significant cytotoxicity against human T-cell lympho-
blast (CCRF-CEM) and breast adenocarcinoma (MCF-7)
cell lines had been reported.[16] Regioisomeric BTAs on
the different nitrogen atoms are difficult to be synthesized
and their anticancer properties remain poorly defined. In
this work, we aimed to design novel molecules that can
biological
properties,
including
antifungal,[1,2]
antibacterial,[3,4] antiviral,[5] and anticancer activities.[6,7]
Protein tyrosine kinases (PTKs) are the crucial class of
oncology drugs targets.[8,9] Most of PTKs inhibitors have
structures of heterocyclic ring with nitrogen-containing
moieties, like BTA as the bioisostere, which competes
with ATP for the adenine-binding region of the protein
tyrosine kinase to inhibit the growth of cancer cells by
blocking the signal transduction of phosphorylation and
inhibiting the abnormal expression of PTKs.[10–12] PTKs
catalyze phosphorylation of tyrosine in many proteins by
the transfer of the γ-phosphoryl group from ATP to regu-
late the fundamental cellular processes, such as prolifera-
tion, differentiation, migration, metabolism, and anti-
apoptotic signaling.[13] As early as 2001, Sarno et al
reported that 4,5,6,7-tetrabromo-1H-benzotriazole could
dock with the ATP-binding pocket of CK2 in different
J Heterocyclic Chem. 2019;1–8.
wileyonlinelibrary.com/journal/jhet
© 2019 Wiley Periodicals, Inc.
1

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LI ET AL.
be used as inhibitors of PTKs with anticancer properties
by fragment-based design strategy. We used BTA as
starting material to synthesize a series of intermediates
with different regioisomeric substituents under
ultrasound-microwave conditions before reacted with
chloro- or methoxylpyridine aldehydes. The antip-
roliferative activities of these target BTA compounds were
screened in epithelial cancer cell line VX2, human lung
cancer cell line A549, human gastric cancer cell lines
MGC and MKN45 in vitro.
condensation as depicted in Figure 1. BTAs were reacted
with chloroacetonitrile to obtain intermediates under
ultrasound-microwave conditions, and then 10 BTA
derivatives were synthesized with 6-chloronicotinaldehyde
and 5-formaldehyde-2-methoxypyridine, respectively. The
ultrasonic-microwave reaction was optimized and carried
out in this study with a ratio of BTA/chloroacetonitrile or
intermediates/aromatic aldehyde of 3:4, microwave 480 W,
and temperature of 75ꢀC. Newly synthesized compounds
were yellow crystalline powder that were in correspondent
with their spectral and elemental analysis data. The struc-
tures of compounds are presented in Figure 1. These com-
pounds are insoluble in water, methanol, or ethanol while
soluble in toluene, DMF, DMSO, or CHCl3. Physical charac-
terization and yields of the target compounds were summa-
rized in Table 1. According to the infrared spectrum of the
intermediate, it was confirmed that the benzotriazole ring
was successfully attached to the acetonitrile group with a
2 | RESULTS AND DISCUSSION
2.1 | Synthesis
Benzotriazole was used as starting material to synthesize
10 target compounds through two steps of substitution and
FIGURE 1 Synthesis of target
compounds

LI ET AL.
3
TABLE 1 Physical characterization and yields of benzotriazole derivatives
Compounds
Molecular formula
C14H8ClN5
Molecular weight
281.70
Color
MP (ꢀC)
144-146
149-151
233-234
138-140
166-167
177-179
209-210
133-136
166-167
177-179
% Yield
30.5
18.8
55.7
23.0
43.7
20.7
60.2
25.6
61.5
25.5
1.1
1.2
1.3
1.4
2.1
2.2
2.3
2.4
2.5
2.6
Yellow
Yellow
Yellow
Yellow
Yellow
Yellow
Yellow
Yellow
Yellow
Yellow
C15H11N5O
277.29
C14H8ClN5
281.70
C15H11N5O
277.29
C14H7Cl2N5
C15H10ClN5O
C14H7Cl2N5
C15H10ClN5O
C14H7Cl2N5
C15H10ClN5O
316.15
311.73
316.15
311.73
316.15
311.73
characteristic stretching around 2200 to 2400 cm−1. Further-
more, the regioisomeric BTA intermediates on the different
nitrogen atoms were separated and characterized before the
condensation reactions. The purification of these isomers
were achieved by flash column chromatography based on
polarity of these compounds. The region-selectivity of differ-
ent intermediates was analyzed based on the changes in dif-
ferent proton signals on the benzene ring in 1H-NMR
spectra.[17,18] When the aromatic hydrogen proton signals
all have signal peaks around 7.50 ppm and the influence of
methylene is small, these compounds are predicted to be a
2-substituted intermediate. In 1-substituted intermediates,
the chemical shifts of 7-position protons are moved to the
high field by the influence of the adjacent methylene group.
In 3-substituted intermediates, the chemical shifts of the
6-position proton and the 7-position proton are on the same
signal peak, suggesting that the two sites are roughly
affected by the influence of chlorine and methylene. Inter-
estingly, region-selective substitution of BTAs at different
positions affected the yields of the final products. The yields
of 2H-1, 2, 3-benzotriazole derivatives or 3H-1,
2, 3-benzotriazole derivatives were generally higher than
the isomers of 1H-1, 2, 3-benzotriazole derivatives presum-
ably due to the less steric effects on 2 or 3-positions compar-
ing to 1 position. Furthermore, yields of BTA derivatives
bonded with aromatic aldehydes were impacted by the dif-
ferent substituents on aromatic rings: chlorine on pyridine
rings has weak electron-withdrawing effect, while methoxyl
group on pyridine rings has strong electron-donating effect.
Electron-withdrawing effect in aldehyde promotes the con-
densation reaction, while the electron-donating group slows
down the reaction. Thus the compounds a methoxyl
substituted pyridine had lower yields than those with a
chlorine substituent. For example, compound 2.5 has the
highest yield with 61.5% and compound 1.2 has lowest yield
with only 18.8%.
2.2 | Antiproliferative activity
Antiproliferative activities of the newly prepared com-
pounds were evaluated against VX2, A549, MGC, and
MKN45 compared with a well-studied PTK inhibitor,
Gefitinib, in vitro. The results were expressed as median
growth inhibitory concentration (IC50) values by using
CCK-8 assays. As showed in the Table 2, 10 target com-
pounds had good inhibition to the proliferation of VX2,
A549, MGC, and MKN45 cell lines, among which MKN45
were more sensitive to target compounds while less sensi-
tive of A549. Compound 2.5 had the better inhibitory effect
on the four cell lines than Gefitinib, while IC50 values of
5.47 0.41 μM on VX2, 5.47 1.11 μM on A549, 4.59
0.14 μM on MGC, 3.04 0.02 μM on MKN45, respec-
tively. Concerning the activity against VX2, compounds 1.1,
1.2, 1.4, 2.1, 2.5 all have better activity than Gefitinib. Com-
pound 2.1 (IC50 of 3.80 0.75 μM) emerged as the most
potent analogue as it was fourfolds more active than
Gefitinib (IC50 of 12.88 2.74 μM). Compounds 1.1, 1.2,
1.3, 2.1, 2.2, 2.5, 2.6 all have much better activity than
Gefitinib against MGC, while compound 2.2 displayed bet-
ter inhibitory effect on MGC with an IC50 value of 3.72
0.11 μM than that of Gefitinib (IC50: 8.14 0.60 μM).
The inhibitory activity of 5-chlorbenzotriazole-series was
slightly better than that of non-substituted BTAs. The inhib-
itory activity of chloronicotin-aldehyde series was better
than that of 6-methoxynicotinaldehyde-series. Furthermore,
the activity of target compounds substituted at 1 and 3 posi-
tions on triazole ring is generally better than that of target
compounds substituted at 2 position. The inhibitory activity
of compounds of 1.1 and 1.3 were stronger than that of tar-
get compounds of 1.2 and 1.4, suggesting that chloro-
substituent on pyridine ring is better than methoxyl
group.[19] On the other hand, the chlorine atom on the ben-
zene ring appears to exert limited effects on the inhibitory

4
LI ET AL.
TABLE 2 Antiproliferation activity
of benzotriazole derivatives on VX2,
A549, MGC, MKN45 cell lines
Corresponding effect on cell lines
Compounds
VX2
A549
MGC
MKN45
measured by CCK-8 assay (unit, μM)
1.1
5.36 1.40
8.23 0.96
56.55 0.21
12.31 1.51
3.80 0.75
30.69 0.59
26.61 2.28
39.94 7.98
5.47 0.41
19.87 1.87
12.88 2.74
10.52 0.25
10.94 3.06
59.41 0.16
27.15 0.53
6.99 0.33
18.20 0.33
21.58 0.99
45.91 0.52
5.47 1.11
12.10 2.14
10.15 0.09
4.65 0.02
5.20 0.15
5.79 0.18
21.77 0.92
6.04 0.21
3.72 0.11
9.78 0.49
18.72 0.85
4.59 0.14
7.29 0.34
8.14 0.60
11.72 0.23
9.28 0.16
5.75 0.05
11.77 0.25
9.27 0.22
3.63 0.08
11.58 0.75
12.55 0.43
3.04 0.02
3.34 0.10
6.94 0.28
1.2
1.3
1.4
2.1
2.2
2.3
2.4
2.5
2.6
Gefitinib
FIGURE 2 Inhibition of cell proliferation of target compounds
activity with a chlrorine on the pyridine (comparing com-
pound 1.1 to 2.1, 1.3 to 2.3). In addition, the inhibitory
activity of compounds 2.5, 2.6 were greater than that of
compound 2.1, 2.2 (Figures 2 and 3). Overall, compound
2.5 showed the strongest inhibitory activities across four cell
lines, which can be further optimized in future studies.
2.3 | Structure activity discussions
The 10 BTA derivatives have stood out as privileged mole-
cules with good antiproliferative properties that were
designed according to the structure of the tyrosine kinase
inhibitor, such as Gefitinib. The BTA and pyridine ring in

LI ET AL.
5
chromatography (TLC) on silica gel G was employed rou-
tinely to follow the course of reactions and check the
purity of products. All melting points of recrystallized
products were measured with a Stuart melting point appa-
1
ratus. All compounds were characterized by IR, H and
13C-NMR, mass spectrometry (MS) and elemental analysis.
IR spectra were recorded in KBr disks on Thermo Nicolet
FIGURE 3 Structure of 2-(5-chloro-1H-benzo[d] [1,2,3]
triazol-1-yl)-3-(6-chloropyridin-3-yl) acrylonitrile, compound 2.5
1
model Avatar 330 FT-IR spectrometer. H and 13C-NMR
spectra were recorded by Avance III, Bruker Biospin at
500/400 MHz in deuterated chloroform or dimethyl sulfox-
ide (CDCl3/DMSO). Mass spectra were measured on
AB3200 QTRAP mass spectrometer.
the target compounds are separated by two carbons. The
phenyl ring of BTA and pyridine ring form π-π interaction
with tyrosine kinase with increasing the degree of conjuga-
tion. The literature survey found that the length of the
spacer carbon atoms has an effect on the cancer activity,
and two to three units are more favorable in preliminary
structure-activity relationship.[20,21] Among them, the tar-
get products at 1 or 3 position substitution on BTA ring
leave two exposed N atoms easier to interact with hydro-
gen bond donors on tyrosine protease.[7] The chlorine
atoms on BTA and pyridine ring are slightly electron-
withdrawing groups, which affect the charge distribution
and space morphology of the target compounds. The bind-
ing mode of target compounds enhances the affinity
between target compound and receptor, which leads to bet-
ter action. The steric hindrance of substituents is smaller,
which is more conducive to the target compound entering
the ATP binding region of tyrosine protein kinase and
competing with ATP to bind receptor to exhibit a higher
inhibition rate. Based on these considerations of structure–
activity relationship, the inhibition of compound 2.5 was
much higher compared with other compounds. Nonethe-
less, further structural optimization is needed in the future,
in addition to investigate its toxicity toward normal cells.
3.2 | 2-(1H-benzo[d] [1,2,3] triazol-1-yl)-
3-(6-chloropyridin-3-yl) acrylonitrile (1.1)
IR (KBr, cm−1): 3046, 2230, 1649, 1608, 1579, 1491, 1454,
1
831, 694; H-NMR(500 MHz, CDCl3, δ): 8.70(1H, d, CH
aromatic), 8.57(1H, dd, CH aromatic), 8.38(1H, d,
CH─), 7.91(2H, m, CH aromatic), 7.62(1H, d, CH aro-
matic), 7.53(2 H, m, CH aromatic); 13C-NMR (400 MHz,
DMSO, δ): 109.7, 111.9, 113.9, 120.6, 125.2, 125.3, 126.2,
127.2, 130.1, 131.7, 125.2, 146.0, 151.6, 152.7 ppm; ESI-
MS: 282.05 (C14H8ClN5, [M + H]+); Anal. Calcd for
C14H8ClN5: C, 59.68; H, 2.86; Cl, 12.58; N, 24.86, found
C59.62; H, 2.85; N, 24.82.
3.3 | 2-(1H-benzo[d] [1,2,3] triazol-1-yl)-
3-(6-methoxypyridin-3-yl) acrylonitrile (1.2)
IR (KBr, cm−1): 2956, 2362, 1610, 1574, 1495, 1452, 1290,
1
1076, 819; H-NMR(500 MHz, CDCl3, δ): 8.52(1H, d, CH
aromatic), 8.44 (1H, dd, CH aromatic), 8.15 (1H, d, CH aro-
matic), 7.91 (1H, d, CH aromatic), 7.88(1 H, s, CH─), 7.65
(1H, m, CH aromatic), 7.50 (1H, m, CH aromatic), 6.93
(1H, d, CH aromatic), 4.04 (3H, s, ─CH3); 13C-NMR
(400 MHz, DMSO, δ): 54.4, 105.5, 111.6, 111.9, 114.7, 120.5,
121.1, 125.9, 129.8, 132.0, 138.6 (two overlapping signals),
145.8, 151.1, 165.8 ppm; ESI-MS: 278.10 (C15H11N5O, [M
+ H]+); Anal. Calcd for C15H12N5O: C, 64.97; H, 4.00; N,
25.26; O, 5.77, found C, 64.92; H, 4.03; N, 25.22.
3 | EXPERIMENTAL
3.1 | Synthesis and characterization
Benzotriazole was used as starting material to synthesize
10 target compounds. The reaction initiated with
27.5 mmol benzotriazole/5-chlorobenzotriazole and 3 mL
chloroacetonitrile in an ultrasonic-microwave reactor
under 75 mL refluxing ethyl acetate at 480 W to afford
intermediates. The BTA intermediates were separated with
different ratios of ethyl acetate/petroleum ether as eluent
by flash column chromatography. Then 3 mmol interme-
diates reacted with 4 mmol 6-chloronicotinaldehyde or
6-methoxynicotinaldehyde in 20 mL absolute toluene by
catalyzing with 1.2 mL triethylamine to get the desired
compounds. The target compounds were separated with
different ratios of ethyl acetate/petroleum ether as eluent
by flash column chromatography. Analytical thin layer
3.4 | 2-(2H-benzo[d] [1,2,3] triazol-2-yl)-
3-(6-chloropyridin-3-yl) acrylonitrile (1.3)
IR (KBr, cm−1): 3017, 2360, 1607, 1572, 1496, 1446,
1
807, 670; H-NMR (500 MHz, CDCl3, δ): 8.74 (1H, d, CH
aromatic), 8.51(1H, dd, CH aromatic), 8.48(1H, d,
CH─), 7.91(2H, m, CH aromatic), 7.52(1H, d, CH aro-
matic), 7.49(2H, m, CH aromatic); 13C-NMR (400 MHz,

6
LI ET AL.
DMSO, δ): 113.1, 114.3, 118.6 (two overlapping signals),
125.3, 126.8 (two overlapping signals), 129.4, 133.6, 139.5,
145.0 (two overlapping signals), 152.1, 153.0 ppm; ESI-
MS: 282.05 (C14H8ClN5, [M + H]+); Anal. Calcd for
C14H8ClN5: C, 59.69; H, 2.86; Cl, 12.58; N, 24.86,
found C, 59.73; H, 2.81; N, 24.82.
for C15H10ClN5O: C, 57.80; H, 3.23; Cl, 11.37; N, 22.47; O,
5.13, found C, 57.85; H, 3.27; N, 22.51.
3.8 | 2-(5-Chloro-2H-benzo[d] [1,2,3]
triazol-2-yl)-3-(6-chloropyridin-3-yl)
acrylonitrile (2.3)
3.5 | 2-(2H-benzo[d] [1,2,3] triazol-2-yl)-
3-(6-methoxypyridin-3-yl) acrylonitrile (1.4)
IR (KBr, cm−1): 2341, 1636, 1558, 1508, 1457, 818, 669;
1H-NMR(500 MHz, DMSO, δ): 8.89 (1H, d, CH aro-
matic), 8.81(1H, d, CH aromatic), 8.51 (1H, dd, CH aro-
matic), 8.26 (1H, d, CH─), 8.13 (1H, dd, CH aromatic),
7.82(1H, d, CH aromatic), 7.61(1H, dd, CH aromatic);
13C-NMR (400 MHz, DMSO, δ): 112.9, 114.1, 117.7,
120.6, 125.4, 126.7, 130.5, 133.9, 134.2, 139.5, 143.5,
145.2, 152.2, 153.2 ppm; ESI-MS: 317.00 (C14H7Cl2N5,
[M + H]+); Anal. Calcd for C14H7N5Cl2: C, 53.19; H,
2.23; Cl, 22.43; N, 22.15; found C, 53.22; H,
2.27; N, 22.13.
IR (KBr, cm−1): 3009, 2258, 1615, 1574, 1496, 1449, 1255,
1
1091, 841; H-NMR(500 MHz, CDCl3, δ): 8.20(1H, d, CH
aromatic), 7.90(2 H, m, CH aromatic), 7.80(1 H, s,
CH─), 7.48(2 H, m, CH aromatic), 6.80(1H, dd,
CHaromatic), 5.80 (1H, d, CH aromatic), 3.93(3H, s,
-CH3); 13C-NMR(400 MHz, DMSO, δ): 53.7, 110.8, 115.1,
115.2, 118.6 (two overlapping signals), 127.8, 128.0 (two
overlapping signals), 138.0, 138.5, 144.4 (two overlapping
signals), 146.5, 164.3 ppm; ESI-MS: 278.10(C15H11N5O,[-
M + H]+); Anal. Calcd for C15H11N5O: C, 64.97; H,
4.00; N, 25.26; O, 5.77, found C, 64.92; H, 3.95; N, 25.22.
3.9 | 2-(5-Chloro-2H-benzo[d] [1,2,3]
triazol-2-yl)-3-(6-methoxypyridin-3-yl)
acrylonitrile (2.4)
3.6 | 2-(6-Chloro-1H-benzo[d] [1,2,3]
triazol-1-yl)-3-(6-chloropyridin-3-yl)
acrylonitrile (2.1)
IR (KBr, cm−1): 3017, 2342, 1607, 1572, 1497, 1446, 1260,
1049, 807, 671; H-NMR (500 MHz, DMSO, δ): 8.20(1H, d,
1
CH aromatic), 8.73(1H, s, CH─), 8,45(1H, dd, CH aro-
matic), 8.22 (1H, d, CH aromatic), 8.10 (1H, d, CH aro-
matic), 7.58 (1H, dd, CH aromatic), 7.09 (1H, d, CH
aromatic), 3.96 (3H, s, -CH3); 13C-NMR(400 MHz, DMSO,
δ): 54.4, 107.5, 112.0, 113.4, 114.8, 125.4, 125.6, 126.8, 131.0,
134.3, 134.5, 144.9, 145.2, 152.1, 164.9 ppm; ESI-MS: 312.05
IR (KBr, cm−1): 3028, 2226, 1652, 1608, 1578, 1507, 1461,
1
814, 651; H-NMR (500 MHz, DMSO, δ): 8.91(1H, d, CH
aromatic), 8.47(1H, dd, CH aromatic), 8.35 (2H, m, CH
aromatic), 8.29 (1H, d, CH─), 7.83(1H, d, CH aromatic),
7.64(1H, dd, CH aromatic); 13C-NMR (400 MHz, DMSO,
δ): 109.3, 113.9, 122.0, 125.3, 126.9, 127.2, 132.6, 135.1,
136.2, 139.5, 139.9, 144.7, 151.7, 152.8 ppm; ESI-MS:
317.00 (C14H7Cl2N5, [M + H]+); Anal. Calcd for
C14H7Cl2N5: C, 53.19; H, 2.23; Cl, 22.43; N, 22.15, found C,
53.22; H, 2.19; N, 22.12.
(C15H10ClN5O,
[M + H]+);
Anal.
Calcd
for
C15H10ClN5O: C, 57.80; H, 3.23; Cl, 11.37; N, 22.47; O, 5.13,
found C, 57.85; H, 3.21; N, 22.52.
3.10 | 2-(5-Chloro-1H-benzo[d] [1,2,3]
triazol-1-yl)-3-(6-chloropyridin-3-yl)
acrylonitrile (2.5)
3.7 | 2-(6-Chloro-1H-benzo[d] [1,2,3]
triazol-1-yl)-3-(6-methoxypyridin-3-yl)
acrylonitrile (2.2)
IR (KBr, cm−1): 2340, 1614, 1573, 1519, 1455, 806, 668;
1H-NMR (500 MHz, DMSO, δ): 8.91 (1H, d, CH aro-
matic), 8.49 (1H, dd, CH aromatic), 8.44 (1H, m, CH aro-
matic), 8.36 (1H, s, CH aromatic), 8.20 (1H, d, CH─),
7.83(2H, d, CH aromatic); 13C-NMR (400 MHz, DMSO,
δ): 109.4, 113.6, 113.8, 119.9, 125.3, 127.1, 130.4, 130.6,
130.7, 135.9, 139.4, 146.7, 151.7, 152.8 ppm; ESI-MS:
317.00 (C14H7Cl2N5, [M + H]+); Anal. Calcd for
C14H7Cl2N5: C, 53.19; H, 2.23; Cl, 22.43; N, 22.15,
found C, 53.15; H, 2.25; N, 22.12.
IR (KBr, cm−1): 3008, 2221, 1683, 1602, 1558, 1507, 1456,
1233, 1048, 796, 668; 1H-NMR (500 MHz, DMSO, δ): 8.73
(1H, d, CH─), 8.42(1H, dd, CH aromatic), 8.27(3H, m,
CH aromatic), 7.62 (1H, dd, CH aromatic), 7.12 (1H, d,
CH aromatic), 3.97 (3H, s, ─CH3); 13C-NMR (400 MHz,
DMSO, δ): 54.4, 106.5, 112.2, 114.2, 114.5，121.4, 127.7，
129.8, 130.7, 131.7, 136.8, 138.3, 145.2, 148.8, 164.8 ppm;
ESI-MS: 312.06 (C15H10ClN5O, [M + H]+); Anal. Calcd

LI ET AL.
7
3.11 | 2-(5-Chloro-1H-benzo[d] [1,2,3]
triazol-1-yl)-3-(6-methoxypyridin-3-yl)
acrylonitrile (2.6)
ACKNOWLEDGMENTS
The authors wish to thank Dr. Zheng Y.X, Dr. Wang
J. and Dr. Xu X.Z for their assistance in cck-8 analysis
and characterization of 1H and 13C-NMR.
IR (KBr, cm−1): 2341, 1597, 1558, 1506, 1457, 1288, 1048,
822, 668; 1H-NMR (500 MHz, DMSO, δ): 8.73 (1H, d,
CH─), 8.43(2H, m, CH aromatic), 8.27 (1 H, s, CH aro-
matic), 8.12 (1H, d, CH aromatic), 7.80(1H, dd, CH aro-
matic), 7.13 (1H, d, CH aromatic), 3.97 (3H, s, -CH3); 13C-
NMR (400 MHz, DMSO, δ): 54.4, 105.3, 112.0, 113.4, 114.6,
119.8, 121.0, 130.2, 130.4, 131.0, 138.6, 139.1, 146.5, 151.2,
165.8 ppm; ESI-MS: 312.06 (C15H10ClN5O, [M + H]+);
Anal. Calcd for C15H10ClN5O: C, 57.80; H, 3.23; Cl,
11.37; N, 22.47; O, 5.13; found C, 57.78; H, 3.20; N, 22.42.
ORCID
Qiujing Li https://orcid.org/0000-0002-6689-9210
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8
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How to cite this article: Li Q, Liu G, Wang N,
Yin H, Li Z. Synthesis and anticancer activity of
benzotriazole derivatives. J Heterocyclic Chem.
2019;1–8. https://doi.org/10.1002/jhet.3859
SUPPORTING INFORMATION
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