Synthesis, molecular modeling, and biological evaluation of 1,2,4-triazole derivatives containing pyridine as potential anti-tumor agents

Article abstract of DOI:10.1007/s00044-012-0306-5

There is an accumulating body of experimental evidences validating focal adhesion kinase (FAK) as a therapeutic target and offering opportunities for anti-tumor drug development. In present study, we sought to synthesize twenty-eight potential FAK inhibit

Full text of DOI:10.1007/s00044-012-0306-5

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res (2013) 22:3193–3203
DOI 10.1007/s00044-012-0306-5
ORIGINAL RESEARCH
Synthesis, molecular modeling, and biological evaluation
of 1,2,4-triazole derivatives containing pyridine as potential
anti-tumor agents
•
Yan-Bin Zhang Wen Liu Yu-Shun Yang
•
•
•
Xiao-Liang Wang Hai-Liang Zhu
•
•
Li-Fei Bai Xiao-Yang Qiu
Received: 27 July 2012 / Accepted: 24 October 2012 / Published online: 15 November 2012
Ó Springer Science+Business Media New York 2012
Abstract There is an accumulating body of experimental
evidences validating focal adhesion kinase (FAK) as a
therapeutic target and offering opportunities for anti-tumor
drug development. In present study, we sought to synthesize
twenty-eight potential FAK inhibitors as anti-tumor agents
based on 1,2,4-triazole skeleton. The bioassay assays dem-
onstrated that compounds 3e and 6j showed the most potent
activity, 3e inhibited the growth of HCT116 and HepG2 cell
lines with IC50 values of 8.17 and 7.04 lM, while compound
6j showed the most potent biological activity against
HCT116 cell line (IC50 = 1.99 lM). Besides, compound 6j
also exhibited significant FAK inhibitory activity
(IC50 = 2.41 lM). The results of flow cytometry and wes-
tern-blot assay demonstrated that compounds 3e and 6j
possessed good anti-proliferative activity. Docking simula-
tions were performed to position compounds 3e and 6j into
the active site of FAK to determine the probable binding
model.
Introduction
Cancer, the second leading cause of death in the world, is
continuing to be a major health problem in developing as
well as undeveloped countries (EI-Azab et al., 2010).
Cancer chemotherapy targeting tumor progression repre-
sents one of the most relevant challenges of chemists and
oncologist. In order to gain new insights into the com-
plexity of the disease, robust screening methods for eval-
uating different natural or synthetic drugs have been
carried out from the science community (Stanton et al.,
2008). Therefore, there is an increasing need for new
therapies, especially those that based on current knowledge
of cancer biology as well as that taking advantage of the
cancer cells phenotype, described by Hanahan and Wein-
berg (2000).
The non-receptor protein tyrosine kinase focal adhesion
kinase (FAK) was discovered almost 15 years ago (Van
Nimwegen and van de Water, 2007). Most studies show
an enhanced expression of FAK mRNA and/or protein in
a variety of human cancers, including squamous cell
carcinoma of the larynx (Aronsohn et al., 2003), invasive
colon and breast tumors (Owens et al., 1995), and
malignant melanoma (Kahana et al., 2002). Activation of
FAK leads to a number of cell biological processes,
including cell attachment, migration, invasion, prolifera-
tion, and survival (Maroesja and Bob, 2007). The human
FAK core promoter contains binding sites for many
transcription factors, including but not limited to NF-jB,
p53, AP-1, AP-2, PU.1, TCF-1, and EGR-1 (Golubovs-
kaya et al., 2004). Inhibition of FAK by anti-sense tech-
niques or antibody-methods results in the onset of
apoptosis and numerous studies have clearly confirmed a
protective role for FAK in apoptosis (Sonoda et al., 2000).
Given the role of FAK in processes important in
Keywords FAK ꢀ 1,2,4-triazole ꢀ Anti-tumor activity ꢀ
Molecular docking
Y.-B. Zhang ꢀ W. Liu ꢀ Y.-S. Yang ꢀ X.-L. Wang ꢀ
H.-L. Zhu (&)
State Key Laboratory of Pharmaceutical Biotechnology, Nanjing
University, 210093 Nanjing, People’s Republic of China
e-mail: zhuhl@nju.edu.cn
L.-F. Bai (&)
School of Life Sciences and Chemistry, Jiangsu Institute
of Education, 210013 Nanjing, People’s Republic of China
X.-Y. Qiu (&)
Department of Chemistry, Shangqiu Normal University,
476000 Shangqiu, People’s Republic of China
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Med Chem Res (2013) 22:3193–3203
tumorigenesis, metastasis, and the link to prominent
oncogenes, FAK might be a promising target in the
ongoing search for an anti-cancer drug (Maroesja and
Bob, 2007).
was refluxed for 2 h in the presence of K2CO3. The solu-
tion was cooled and compound 4 appeared. Then, com-
pound 5 was prepared by treatment of 4 with hydrazine
hydrate (85 %) in ethanol. Finally, fourteen 1,2,4-triazole
derivatives (6a–6n) were collected by stirring in ethanol of
5 with different benzaldehyde compounds as shown in
Scheme 1.
The chemistry of N-bridged heterocycles derived from
1,2,4-triazole has received considerable attention in recent
years due to their usefulness in different areas of biological
activities and as industrial intermediates (Bhat et al., 2009).
1,2,4-triazole derivatives are known to exhibit anti-micro-
bial (Ashok and Holla, 2007; Prasad et al., 2009), anti-
tubercular (Walczak et al., 2004), anti-cancer (Sztanke and
Tuzimski, 2008; Romagnoli and Baraldi, 2010), anti-con-
vulsant (Amir and Shikha, 2004), anti-inflammatory, and
analgesic properties (Almasirad et al., 2004). 1,2,4-triazole
nucleus has been incorporated into a wide variety of ther-
apeutically interesting drug candidates including H1/H2
histamine receptor blockers, cholinesterase active agents,
CNS stimulants, anti-anxiety, and sedatives (Schreier and
Helv, 1976), anti-mycotic activity such as fluconazole,
itraconazole, and voriconazole (Budavari et al., 1996;
Haber, 2001). Some 1,3,4-thiadiazole derivatives have
been reported as potential FAK inhibitors (Juan et al.,
2011). Pyridine was used to replace benzo[b]dioxin mainly
according to the result of CADD (computer assistant drug
design) method. The result indicated that a smaller moiety
with higher electronic density might improve the bioac-
tivity. Herein, in continuation to extend our research on
anti-tumor compounds with FAK structure inhibitory
activity, in the present study we sought to synthesis two
series of 1,2,4-triazole derivatives containing pyridine as
anti-tumor agents. Biological evaluation indicated that
some of the synthesized compounds were potent inhibitors
of FAK structure.
All of the synthetic compounds gave satisfactory ana-
lytical and spectroscopic data, which were in full accor-
dance with their depicted structures. Furthermore, the
crystal data, data collection, and refinement parameter for
compound are listed in Table 1, and Fig. 1 gives a per-
spective view of this compound together with the atomic
labeling system. The structure was solved by direct meth-
ods and refined on F2 by full-matrix least-squares methods
using SHELX-97 (Sheldrick, 1997).
Biological activity
All the synthesized derivatives 3a–3n and 6a–6n were
evaluated for their anti-proliferative activity against MCF7
(human breast cancer cells), HCT116 (human colorectal
cancer cells), and HepG2 (human hepatoma cells) cell
lines. The results were summarized in Table 2. As illus-
trated in Table 2, compounds 3a–3n and 6a–6n showed all
certain activities against the three tumor cells as compared
with staurosporine. For the studied compounds, we
observed that compounds (3e, 6j) showed potent anti-
cancer activities against MCF7, HCT116, and HepG2 cell
lines. It was obvious that compound 6j exhibited best
activity against HCT116 cells with the IC50 value of
1.99 lM, which was even better than the reference drug
staurosporine (IC50 = 13.20 lM).
Structure–activity relationships (SARs) in these 1,2,
4-triazole derivatives demonstrated that compounds having
halogen atom substituent exhibited high activity. Mean-
while, a comparison of the substitution on benzene ring
was demonstrated as follows (3a–3n): when the com-
pounds were bromine-substituted derivatives, the potency
order was meta [ para [ ortho (3c, 3e, 3h), and when the
compounds were chlorine-substituted derivatives, the
potency order was also meta [ para [ ortho (3b, 3d, 3g).
Interestingly, among the electron-withdrawing substituents
compounds, the potency order was almost bro-
mine [ chlorine [ fluorine [ nitro. Another comparison
of the substitution on benzene ring was demonstrated as
follows (6a–6n): compounds with electron-donating sub-
stituents at para-position at A-ring had better inhibitory
activity than that with electron-withdrawing substituents at
the same position. When the compounds were fluorine-
substituted derivatives, the potency order was para [
ortho [ meta (6g, 6h, 6i), among the electron-withdrawing
substituents compounds, the potency order was
Results and discussion
Chemistry
In this study, twenty-eight triazole derivatives containing
pyridine were synthesized. The synthetic route of com-
pounds 3a–3n and 6a–6n was shown in Scheme 1. It was
prepared in six steps. First, a mixture of corresponding
isoniazid and phenyl isothiocyanate was refluxed in ethanol
for 30 min. The solution was cooled and compound 1 was
precipitated. A solution of NaOH (2 N) containing 1 was
stirred under refluxing for 30 min, yielding the desired
compound 2 according to the reported procedure. Then,
fourteen 1,2,4-triazole derivatives (3a–3n) were prepared
by refluxing in anhydrous acetonitrile of 2 with different
benzyl bromide compounds as shown in Scheme 1
according to the literature. To a solution of compound 2 in
acetone, ethyl bromoacetate was added and the mixture
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Med Chem Res (2013) 22:3193–3203
3195
O
C
O
C
Scheme 1 General synthesis of
compounds (3a–3n and 6a–6n).
Reagents and conditions:
a Phenyl isothiocyanate,
ethanol, reflux, 30 min;
H
N
H
N
NH2
N
H
a
b
N
H
N
S
N
1
b NaOH (2 N), reflux, 30 min.
c Reflux, acetonitrile.
N
R1
N
N
N
R4
d Bromoacetate, K2CO3,
acetone, reflux, 2 h. e Hydrazine
hydrate (85 %), ethanol, rt,
8–12 h; f Ethanol, rt
SH
S
R2
R3
c
Br
N
N
+
R3
N
N
R4
R1
R2
2
3
d
O
O
N
N
R5
N
N
S
S
OHC
R6
R7
OCH3
HN
NH2
N
e
N
+
N
N
R8
4
5
O
N
N
R8
S
HN
N
f
N
A
N
R7
R5
R6
6
R1=F, R2=R3=R4=H
R1=Cl, R2=R3=R4=H
R1=Br, R2=R3=R4=H
R2=Cl, R1=R3=R4=H
R2=Br, R1=R3=R4=H
R3=F, R1=R2=R4=H
R1=R2=R3=R4=H
R7=Cl, R5=R6=R8=H
R7=F, R5=R6=R8=H
R5=F, R6=R7=R8=H
R6=F, R5=R7=R8=H
3a
3b
3c
3d
3e
3f
3k
3l
6f
R1=NO2, R2=R3=R4=H
R2= NO2, R1=R3=R4=H
R3= NO2, R1=R2=R4=H
6g
6h
6i
3m
3n
R5=OH, R6=R7=H, R8=Br
R5=OH, R6=R7=R8=H
R5=OH,R6=R8=Br, R7=H
R5=R8=H, R6=R7=OH
A=C6H5-C=C-
6j
R5=R6=R7=R8=H
6a
6k
3g R3=Cl, R1=R2=R4=H
6b R7=OCH3, R5=R6=R8=H 6l
R3=Br, R1=R2=R4=H
R1=R3=F, R2=R4=H
R1=R4=F, R2=R3=H
R7=NO2, R5=R6=R8=H
R7=OH, R5=R6=R8=H
R7=Br, R5=R6=R8=H
3h
3i
6c
6d
6e
6m
6n
3j
bromine [ chlorine [ fluorine [ nitro. Compounds with
hydroxy at ortho-position had better inhibitory activity (6j,
6k, 6l). Overall, series 6a–6n had better activities than
series 3a–3n. That might be caused by the additional
hydrazone structure, and it will also be mentioned in the
SARs study part.
compounds were probably correlated to their FAK inhibi-
tory activities.
Apoptosis assay
Apoptosis is an essential mechanism used to eliminate
activated HCT116 cells during the shutdown process of
excess immune responses and maintain proper immune
homeostasis, while deficient apoptosis of the activated
HCT116 cell is associated with a wide variety of immune
disorders. We detected the mechanism of compounds 3e
and 6j (Fig. 2) inhibition effects by flow cytometry, and
found that the compounds could induce the apoptosis of
activated HCT116 cells in a dose- dependent manner. The
result indicated that compounds 3e and 6j induced apop-
tosis of anti-tumor stimulated HCT116 cells.
In addition, we also selected the top 7 compounds which
had better anti-proliferative activity to test their FAK
inhibitory activity against HepG2 cell line. The results
were summarized in Table 3. Most of the tested com-
pounds displayed potent FAK inhibitory. Among them,
compound 6j showed the most potent inhibitory with IC50
of 2.41 lM. The results of FAK inhibitory activity of the
tested compounds were corresponding to the structure
relationships of their anti-tumor activities. This demon-
strated that the potent anti-tumor activities of the synthetic
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Table 1 Crystallographical and experimental data for compound 3g
Table 2 Anti-proliferative activity of the synthesized compounds
(3a–3n and 6a–6n)
Crystal parameters
Compound 3g
Compounds
IC50 (lM)
Formula
C20H15ClN4S
378.07
MCF7
HCT116
HepG2
Formula weight
Crystal size (mm)
a (°)
b (°)
c (°)
0.2 9 0.1 9 0.1
108.585(3)
94.382(4)
91.66(4)
5.765(2)
12.280(5)
13.427(5)
896.8(6)
4
3a
[50
11.97
13.85
20.15
9.82
[50
3b
18.07
16.06
10.54
10.04
21.05
13.27
12.55
14.10
[50
15.70
17.11
11.17
7.04
3c
3d
˚
a (A)
3e
8.17
˚
b (A)
3f
12.80
10.70
11.09
11.39
22.10
16.91
[50
15.62
16.94
12.88
10.15
[50
˚
c (A)
3g
˚
V (A)
3h
Z
3i
Calculated density
(Mg cm-3
1.751
3j
)
3k
22.38
[50
12.91
[50
Absorption coefficient
(mm-1
1.385
3l
)
3m
18.29
22.84
23.64
23.73
15.28
[50
19.81
23.82
14.68
19.23
10.18
[50
21.96
[50
F (0 0 0)
h range (°)
hkl limits
470
3n
2.73-26.00
6a
18.60
27.07
15.89
[50
-7 B h B 7, -15 B k B 15,
-16 B l B 16
6b
6c
Reflections collected/unique
Data/restraints/parameters
R1/wR2 [I [ 2r(I)]
R1/wR2 (all data)
6,642/3,452 [Rint = 0.0256]
3,452/0/235
6d
6e
7.04
4.50
5.99
0.0565/0.1762
0.0829/0.1982
0.834
6f
9.93
5.74
10.95
9.33
6g
12.28
22.43
[50
7.04
GOF
6h
18.04
[50
15.17
24.36
4.39
6i
6j
6.46
1.99
6k
10.04
8.44
7.06
11.26
7.59
6l
4.50
6m
19.73
11.17
16.06
14.68
6.15
16.89
8.28
6n
Staurosporine
13.20
9.04
Molecular docking study
To gain better understanding on the potency of the syn-
thesized compounds and guide further SARs studies. The
FAK-7PY protein–ligand complex crystal structure (PDB
ID: 2ETM) was chosen as the template to compare the
docking mode between our compounds and FAK. The
molecular docking was performed by embedding potent
inhibitors 3e and 6j into binding site of FAK. All docking
runs were applied using Discovery Studio3.1 (DS. 3.1). In
the binding mode, compound 3e (Fig. 4) is bound to FAK
via two p-cation interactions. One p-cation interaction
Fig. 1 Molecule structure of compound 3g
Western-blot assay
In an effort to study the preliminary mechanism of the
compounds with potent inhibitory activity, the western-blot
experiment was performed to explore the effect of com-
pounds 3e and 6j. The western-blot results were summa-
rized in Fig. 3a and b, confirming compounds 3e and 6j’s
inhibitory activities. The result indicated that compound 6j
showed excellent inhibitory activity.
˚
(4.76 A) is formed between LYS 454 and benzene ring
˚
while the other (6.19 A) is formed between LYS 454 and
triazole ring. In addition, compound 6j (Fig. 4) is bound to
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Med Chem Res (2013) 22:3193–3203
3197
Table 3 FAK inhibitory activity of selected compounds
Compound
FAK (IC50, lM)
3e
14.64
17.38
9.17
3i
6e
6f
11.98
17.40
2.41
6g
6j
6n
19.38
3.12
Staurosporine
FAK via one hydrogen bond and two p-cation interactions.
Imino group on the acylhydrazone group and oxygen atom
of CYS 502 form the hydrogen bond (distance: N–HꢀꢀꢀO:
˚
2.35 A, angle: 119.98°). Analogously, one p-cation inter-
˚
action (4.79 A) is formed between LYS 454 and benzene
˚
ring while the other (6.61 A) is formed between LYS 454
Fig. 3 a Compounds 3e and 6j were examined by western blotting.
Data are representative of three independent experiments. b Com-
pounds 6j were examined by western blotting. Data are representative
of three independent experiments
and triazole ring. These results, along with the data of
inhibitory activity assay indicated that compounds 3e and
6j would be a kind of potential inhibitors.
As the combinations shown in Fig. 5, Compound 2p
(Juan et al., 2011) is nicely bound to FAK with two
interactions. The amino hydrogen atom of LYS 454 forms
a hydrogen bond with the oxygen atoms of 1,4-dioxane
group. The nitrogen atom of amide group forms another
hydrogen bond with the oxygen atom of CYS 502.
¨
Fig. 2 HCT116 cells isolated from naıve mice were cultured with anti-cancer and various concentrations of 3e and 6j for 24 h. Cells were
stained by Annexin VeFITC/PI and apoptosis was analyzed by flow cytometry
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Med Chem Res (2013) 22:3193–3203
Fig. 4 Molecular docking modeling of compounds 3e and 6j with
FAK: for clarity, only interacting residues are displayed. Left 3D model
of the interaction between compounds 3e, 6j, and the 2ETM binding
site. The H-bond (green lines) is displayed as dotted lines, and the
p-cation interactions are shown as orange lines. Right 2D model of the
interaction between compound 3e, 6j, and the 2ETM binding site. The
H-bond (blue arrows) is displayed as dotted arrows, and the p-cation
interactions are shown as orange lines (Color figure online)
Fig. 5 2D model of the interaction between compounds 2p, 7PY and the 2ETM binding site. The H-bond is displayed as dotted arrows, and the
p-cation interactions are shown as orange lines (Color figure online)
Compound 7YP is also nicely bound to FAK with three
interactions. The amino hydrogen atom of LYS 454 forms
a hydrogen bond with the nitrogen atom of pyridine group.
Also the pyridine ring of compound 7PY forms a p-cation
interaction with LYS 454. Besides, the pyrrole ring of
compound 7PY forms a p-cation interaction with ARG
550. This insures the binding affinity and results in an
increased FAK inhibitory activity.
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Med Chem Res (2013) 22:3193–3203
3199
In summary, the chosen 1,2,4-triazole derivatives con-
taining pyridine are nicely combined to the FAK. LYS 454
and CYS 502 play an important part in the combination of
the receptor and ligand.
of drug as 100, 30, 10, 3 lM. The remaining ATP in
solution was then quantified utilizing the Kinase-Glo-
luminescence kit (Promega).
Apoptosis assay
Conclusions
HCT116 cells were treated with various concentrations of
compounds 3e and 6j for 24 h and then stained with both
Annexin V-FITC (fluorescein isothiocyanate) and propidi-
um iodide (PI). Then samples were analyzed by FACSCal-
ibur flow cytometer (Becton–Dickinson, SanJose, CA).
Two series of 1,2,4-triazole derivatives containing pyridine
have been synthesized and evaluated for their anti-tumor
activities. Compound 3e and 6j demonstrated the most potent
inhibitory activity. 3e inhibited the growth of the three cell
lines with IC50 values range from 7.04 to 10.04 lM, while 6j
inhibited the growth of the three cell lines with IC50 values
range from 1.99 to 6.46 lM. Besides 6j also inhibited the
activity of FAK with IC50 of 2.41 lM, which was compa-
rable to the positive control staurosporine. In order to gain
deeper understanding of the SARs observed at the FAK,
molecular docking of the most potent inhibitor 3e and 6j into
the binding site of FAK was performed on the binding model
based on the FAK complex structure. Analysis of the com-
pound 6j’s binding conformation demonstrated that com-
pound 6j was stabilized by hydrogen bonding interaction
with CYS502. Apoptosis assay and western-blot results
showed the compound 6j was a potential anti-tumor agent.
Western-blot analysis
After incubation, cells were washed with PBS and lysed
using lysis buffer (30 mm Tris, pH 7.5, 150 mm NaCl, 1 mm
phenylmethylsulfonyl fluoride, 1 mm Na3VO4, 1 % Nonidet
P-40, 10 % glycerol, and phosphatase and protease inhibi-
tors). After centrifugation at 10,0009g for 10 min, the pro-
tein content of the supernatant was determined by a BCATM
protein assay kit (Pierce, Rockford, IL, USA). The protein
lysates were separated by 10 % SDS-PAGE and subse-
quently electrotransferred onto a polyvinylidene difluoride
membrane (Millipore, Bedford, MA, USA). The membrane
was blocked with 5 % non-fat milk for 2 h at room tem-
perature. The blocked membrane was probed with the indi-
cated primary antibodies overnight at 4 °C, and then
incubated with a horse radish peroxidase-coupled secondary
antibody. Detection was performed using a LumiGLO
chemiluminescent substrate system (KPL, Guildford, UK).
Experimental
Cell proliferation assay
The anti-tumor activities of compounds 3a–3n and 6a–6n
were determined using a standard (MTT)-based colori-
metric assay (Sigma). Seed 104 cells per well into 96-well
plates, incubate at 37 °C, 5 % CO2 for 24 h. Then add
100 lL a series concentration of drug-containing medium
into wells to maintain the final concentration of drug as
100, 30, 10, 3 lM. One concentration should be triplicated.
And staurosporine was used for the positive control. After
48 h, cell survival was determined by the addition of an
MTT solution (25 lL of 5 mg mL-1 MTT in PBS). After
4 h, discard the medium and add 100 lL DMSO; the plates
were votexed for 10 min to make complete dissolution.
Optical absorbance was measured at 490 nm.
Molecular docking modeling
Molecular docking of compounds into the 3D FAK com-
plex structure (PDB code: 2ETM) was carried out using the
Discovery Studio (version 3.1) as implemented through the
graphical user interface DS-CDocker protocol.
The three-dimensional structures of the aforementioned
compounds were constructed using Chem 3D ultra 12.0
software [Chemical Structure Drawing Standard; Cam-
bridge Soft corporation, USA (2010)], then they were ener-
getically minimized using MMFF94 with 5000 iterations and
minimum RMS gradient of 0.10. The crystal structures of
FAK complex were retrieved from the RCSB Protein Data
Bank (http://www.rcsb.org/pdb/home/home.do). All bound
water and ligands were eliminated from the protein and the
polar hydrogen was added. The whole FAK complex was
defined as a receptor and the site sphere was selected based
on the ligand binding location of 7PY, then the 7PY mole-
cule was removed and 3e and 6j were placed during the
molecular docking procedure. Types of interactions of the
docked protein with ligand were analyzed after the end of
molecular docking.
FAK inhibitory assay
Seven 1,2,4-triazole derivatives containing pyridine were
tested in a search for small molecule inhibitors of FAK. In
a typical study, human recombinant full-length FAK was
incubated in kinase buffer containing ATP and the sub-
strate for 4 h at room temperature with or without the
presence of the triazole derivatives, the final concentration
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Chemistry
4-[5-(2-Bromo-benzylsulfanyl)-4-phenyl-4H-[1,2,4]triazol-
3-yl]-pyridine (3c)
All the NMR spectra were recorded on a Bruker DPX 300
1
model Spectrometer in CDCl3. Chemical shifts (d) for H
Yellow solid, yield 79 %, mp: 145–146 °C. 1H NMR
(300 MHz, CDCl3): 4.64 (s, 2H); 7.20–7.31 (m, 2H);
7.61–7.70 (m, 5H); 7.80 (s, 2H); 8.18 (d, J = 8.76 Hz,
2H); 8.60 (s, 2H). ESI–MS 423.0 (C20H16BrN4S [M?H]?).
Anal. Calcd for C20H15BrN4S: C, 56.74; H, 3.57; N, 13.23.
Found: C, 56.54; H, 3.56; N, 13.20.
NMR spectra were reported in parts per million to residual
solvent protons. Melting points were measured on a Boe-
tius micro melting point apparatus. The ESI–MS spectra
were recorded on a Mariner System 5304 Mass spec-
trometer. All chemicals and reagents used in current study
were of analytical grade. TLC was run on the silica gel
coated aluminum sheets (Silica Gel 60 GF254, E. Merk,
Germany) and visualized in UV light (254 nm).
4-[5-(3-Chloro-benzylsulfanyl)-4-phenyl-4H-
[1,2,4]triazol-3-yl]-pyridine (3d)
General procedure for synthesis of the target
Yellow solid, yield 81 %, mp: 149–150 °C. 1H NMR
(300 MHz, CDCl3): 4.47 (s, 2H); 7.13 (d, J = 6.30 Hz,
2H); 7.18–7.28 (m, 3H); 7.33 (d, J = 6.76 Hz, 3H);
7.49–7.60 (m, 3H); 8.54 (d, J = 6.06 Hz, 2H). ESI–MS
379.1 (C20H16ClN4S [M?H]?). Anal. Calcd for
C20H15ClN4S: C, 63.40; H, 3.99; N, 14.79. Found: C,
63.21; H, 4.00; N, 14.84.
compounds (3a–3n) and (6a–6n)
To a solution of compound 2 (1 mmol) in acetonitrile, the
corresponding benzyl bromide compounds (1 mmol) were
addedandthemixturewasstirredunderrefluxfor4–8 hinthe
presence of NaOH (2 mmol). Then, the solvent was removed
under reduced pressure and a solid obtained. The solid was
recrystallized from acetonitrile to afford compounds 3a–3n.
To a stirred solution of compound 5 (1 mmol) and the
corresponding benzaldehyde compounds (1 mmol) in eth-
anol (15 mL), water (1 mL) was added followed by drop-
wise addition of glacial acetic acid (0.2 mL). The resulting
mixture was stirred at room temperature until the target
product precipitate from the solvent, which was collected
using suction filtration and dried. The solid was recrystal-
lized from acetonitrile to afford compounds 6a–6n.
4-[5-(3-Bromo-benzylsulfanyl)-4-phenyl-4H-[1,2,4]triazol-
3-yl]-pyridine (3e)
White solid, yield 85 %, mp: 155–156 °C. 1H NMR
(300 MHz, CDCl3): 4.51 (s, 2H); 7.14–7.26 (m, 3H); 7.37
(dd, J1 = 7.68 Hz, J2 = 11.70 Hz, 2H); 7.54 (s, 1H);
7.59–7.68 (m, 3H); 7.86 (d, J = 6.76 Hz, 2H); 8.75 (d,
J = 6.76 Hz, 2H). ESI–MS 423.0 (C20H16BrN4S
[M?H]?). Anal. Calcd for C20H15BrN4S: C, 56.74; H,
3.57; N, 13.23. Found: C, 56.56; H, 3.58; N, 13.27.
4-(5-((2-Fluorobenzyl)thio)-4-phenyl-4H-1,2,4-triazol-3-
yl)pyridine (3a)
4-(5-((4-Fluorobenzyl)thio)-4-phenyl-4H-1,2,4-triazol-3-
yl)pyridine (3f)
Yellow solid, yield 83 %, mp: 173–174 °C. 1H NMR
(300 MHz, CDCl3): 4.62 (s, 2H); 7.02–7.12 (m, 2H); 7.24 (d,
J = 11.52 Hz, 1H); 7.31 (d, J = 5.28 Hz, 1H); 7.51 (t,
J = 7.78 Hz, 1H); 7.61–7.70 (m, 4H); 7.92 (d, J = 6.24 Hz,
2H); 8.64 (d, J = 6.22 Hz, 2H). ESI–MS 363.1
(C20H16FN4S [M?H]?). Anal. Calcd for C20H15FN4S: C,
66.28; H, 4.17; N, 15.46. Found: C, 66.09; H, 4.16; N, 15.50.
Yellow solid, yield 77 %, mp: 142–143 °C. 1H NMR
(300 MHz, CDCl3): 4.53 (s, 2H); 6.91–7.00 (m, 2H); 7.14
(m, 2H); 7.35–7.40 (m, 4H); 7.50–7.61 (m, 3H); 8.55 (d,
J = 6 Hz, 2H). ESI–MS 363.1 (C20H16FN4S [M?H]?).
Anal. Calcd for C20H15FN4S: C, 66.28; H, 4.17; N, 15.46.
Found: C, 66.06; H, 4.18; N, 15.51.
4-[5-(2-Chloro-benzylsulfanyl)-4-phenyl-4H-
[1,2,4]triazol-3-yl]-pyridine (3b)
4-[5-(4-Chloro-benzylsulfanyl)-4-phenyl-4H-
[1,2,4]triazol-3-yl]-pyridine (3g)
Yellow solid, yield 87 %, mp: 139–140 °C. 1H NMR
(300 MHz, CDCl3): 4.63 (s, 2H); 7.12 (d, J = 8.26 Hz, 2H);
7.19–7.23 (m, 2H); 7.28 (t, J = 3.78 Hz, 2H); 7.33–7.37 (m,
1H); 7.49 (t, J = 6.96 Hz, 3H); 7.55–7.59 (m, 1H); 8.53 (d,
J = 6.02 Hz, 2H). ESI–MS 379.1 (C20H16ClN4S [M?H]?).
Anal. Calcd for C20H15ClN4S: C, 63.40; H, 3.99; N, 14.79.
Found: C, 63.61; H, 4.00; N, 14.75.
White solid, yield 84 %, mp: 143–144 °C. 1H NMR
(300 MHz, CDCl3): 4.47 (s, 2H); 7.14 (d, J = 7.22 Hz,
2H); 7.26 (d, J = 9.34 Hz, 2H); 7.31 (t, J = 7.01 Hz, 4H);
7.49–7.56 (m, 3H); 8.54 (d, J = 6.04 Hz, 2H). ESI–MS
379.1 (C20H16ClN4S [M?H]?). Anal. Calcd for
C20H15ClN4S: C, 63.40; H, 3.99; N, 14.79. Found: C,
63.24; H, 3.98; N, 14.74.
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Med Chem Res (2013) 22:3193–3203
3201
4-[5-(4-Bromo-benzylsulfanyl)-4-phenyl-4H-[1,2,4]triazol-
3-yl]-pyridine (3h)
4-[5-(3-Nitro-benzylsulfanyl)-4-phenyl-4H-[1,2,4]triazol-
3-yl]-pyridine (3m)
Yellow solid, yield 78 %, mp: 144–145 °C. 1H NMR
(300 MHz, CDCl3): 4.46 (s, 2H); 7.14 (d, J = 8.14 Hz,
2H); 7.24 (s, 1H); 7.27 (s, 1H); 7.37 (s, 1H); 7.40 (d,
J = 9.50 Hz, 3H); 7.49–7.60 (m, 3H); 8.55 (s, 2H). ESI–
MS 423.0 (C20H16BrN4S [M?H]?). Anal. Calcd for
C20H15BrN4S: C, 56.74; H, 3.57; N, 13.23. Found: C,
56.55; H, 3.58; N, 13.28.
Yellow solid, yield 80 %, mp: 154–155 °C. 1H NMR
(300 MHz, CDCl3): 4.59 (s, 2H); 7.17 (d, J = 7.50 Hz,
2H); 7.28 (t, J = 5.86 Hz, 2H); 7.46–7.58 (m, 4H); 7.81 (d,
J = 7.68, 1H); 8.12 (d, J = 7.32 Hz, 1H); 8.25 (s, 1H);
8.53 (d, J = 5.68 Hz, 2H). ESI–MS 390.1 (C20H16N5O2S
[M?H]?). Anal. Calcd for C20H15N5O2S: C, 61.68; H,
3.88; N, 17.98. Found: C, 61.82; H, 3.89; N, 18.04.
4-(5-((2,4-Difluorobenzyl)thio)-4-phenyl-4H-1,2,4-triazol-
3-yl)pyridine (3i)
4-[5-(4-Nitro-benzylsulfanyl)-4-phenyl-4H-[1,2,4]triazol-
3-yl]-pyridine (3n)
Yellow solid, yield 80 %, mp: 172–173 °C. 1H NMR
(300 MHz, CDCl3): 4.55 (s, 2H); 6.80–6.83 (m, 2H);
7.20–7.23 (m, 2H); 7.58–7.64 (m, 6H); 8.57 (d,
J = 6.42 Hz, 2H). ESI–MS 381.1 (C20H15F2N4S
[M?H]?). Anal. Calcd for C20H14F2N4S: C, 63.15; H,
3.71; N, 14.73. Found: C, 63.33; H, 3.72; N, 14.78.
Yellow solid, yield 77 %, mp: 90–91 °C. 1H NMR
(300 MHz, CDCl3): 4.59 (s, 2H); 7.18 (d, J = 6.96 Hz,
2H); 7.38 (d, J = 5.86 Hz, 2H); 7.53–7.63 (m, 5H); 8.16
(d, J = 8.80 Hz, 2H); 8.55 (d, J = 6.04 Hz, 2H). ESI–MS
390.1 (C20H16N5O2S [M?H]?). Anal. Calcd for
C20H15N5O2S: C, 61.68; H, 3.88; N, 17.98. Found: C,
61.51; H, 3.89; N, 18.03.
4-[5-(2,6-Difluoro-benzylsulfanyl)-4-phenyl-4H-
[1,2,4]triazol-3-yl]-pyridine (3j)
(E)-N0-benzylidene-2-((4-phenyl-5-(pyridin-4-yl)-4H-1,2,4-
triazol-3-yl)thio)acetohydrazide (6a)
Yellow solid, yield 82 %, mp: 142–143 °C. 1H NMR
(300 MHz, CDCl3): 4.62 (s, 2H); 7.01–7.11 (m, 2H);
7.21–7.29 (m, 2H); 7.51 (t, J = 7.68 Hz, 2H); 7.59–7.68
(m, 2H); 7.87 (s, 2H); 8.62 (d, J = 6.76 Hz, 2H). ESI–MS
381.1 (C20H15F2N4S [M?H]?). Anal. Calcd for
C20H14F2N4S: C, 63.15; H, 3.71; N, 14.73. Found: C,
63.37; H, 3.70; N, 14.78.
White powder, yield 87 %; mp: 252–253 °C. 1H NMR
(300 MHz, DMSO): 4.52 (s, 2H); 7.30 (s, 2H); 7.45 (s,
1H); 7.50–7.52 (m, 4H); 7.61–7.62 (m, 3H); 7.69–7.73 (m,
2H); 8.01 (s, 1H); 8.57 (s, 2H); 11.73 (s, 1H). ESI–MS:
415.1 (C22H19N6OS, [M?H]?). Anal. Calcd for
C22H18N6OS: C, 63.75; H, 4.38; N, 20.28 %. Found: C,
63.58; H, 4.39; N, 20.22 %.
4-(5-(Benzylthio)-4-phenyl-4H-1,2,4-triazol-3-yl)pyridine
(3k)
(E)-N0-(4-methoxybenzylidene)-2-((4-phenyl-5-(pyridin-4-
yl)-4H-1,2,4-triazol-3-yl)thio)acetohydrazide (6b)
White solid, yield 86 %, mp: 155–156 °C. 1H NMR
(300 MHz, CDCl3): 4.53 (s, 2H); 7.12–7.14 (m, 2H);
7.27–7.30 (m, 3H); 7.40–7.42 (m, 4H); 7.49–7.57 (m, 3H);
8.54 (d, J = 6.18 Hz, 2H). ESI–MS 345.1 (C20H17N4S
[M?H]?). Anal. Calcd for C20H16N4S: C, 69.74; H, 4.68;
N, 16.27. Found: C, 69.50; H, 4.69; N, 16.33.
White powder, yield 90 %; mp: 228–229 °C. 1H NMR
(300 MHz, DMSO): 3.79 (s, 3H); 4.50 (s, 2H); 6.97–6.99
(m, 2H); 7.26–7.28 (m, 2H); 7.48 (s, 2H); 7.59–7.64 (m,
5H); 7.94 (s, 1H); 8.54–8.55 (m, 2H); 11.56 (s, 1H). ESI–
MS: 445.1 (C23H21N6O2S, [M?H]?). Anal. Calcd for
C23H20N6O2S: C, 62.15; H, 4.54; N, 18.91. Found: C,
62.35; H, 4.53; N, 18.85.
4-[5-(2-Nitro-benzylsulfanyl)-4-phenyl-4H-[1,2,4]triazol-
3-yl]-pyridine (3l)
(E)-N0-(4-nitrobenzylidene)-2-((4-phenyl-5-(pyridin-4-yl)-
4H-1,2,4-triazol-3-yl)thio)acetohydrazide (6c)
Yellow solid, yield 82 %, mp: 141 °C. 1H NMR
(300 MHz, CDCl3): 4.89 (s, 2H); 7.14 (d, J = 6.78 Hz,
2H); 7.33 (d, J = 4.66 Hz, 2H); 7.43–7.62 (m, 5H); 7.94
(d, J = 7.68 Hz, 1H); 8.08 (d, J = 8.22 Hz, 1H); 8.53 (d,
J = 6.24 Hz, 2H). ESI–MS 390.1 (C20H16N2O2S
[M?H]?). Anal. Calcd for C20H15N2O2S: C, 61.68; H,
3.88; N, 17.98. Found: C, 61.48, H, 3.89; N, 18.04.
White powder, yield 91 %; mp: 248–249 °C. 1H NMR
(300 MHz, DMSO): 4.56 (s, 2H); 7.26 (s, 2H); 7.48 (s,
2H); 7.59–7.60 (m, 3H); 7.92–7.97 (m, 2H); 8.11–8.31 (m,
3H); 8.54 (s, 2H); 11.95 (s, 1H). ESI–MS: 460.1
(C22H18N7O3S, [M?H]?). Anal. Calcd for C22H17N7O3S:
123

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Med Chem Res (2013) 22:3193–3203
C, 57.51; H, 3.73; N, 21.34. Found: C, 57.35; H, 3.72; N,
21.41.
Calcd for C22H17FN6OS: C, 61.10; H, 3.96; N, 19.43.
Found: C, 61.27; H, 3.95; N, 19.37.
(E)-N0-(4-hydroxybenzylidene)-2-((4-phenyl-5-(pyridin-4-
yl)-4H-1,2,4-triazol-3-yl)thio)acetohydrazide (6d)
(E)-N0-(3-fluorobenzylidene)-2-((4-phenyl-5-(pyridin-4-yl)-
4H-1,2,4-triazol-3-yl)thio)acetohydrazide (6i)
White powder, yield 93 %; mp: 284–285 °C. 1H NMR
(300 MHz, DMSO): 4.48 (s, 2H); 6.79 (d, J = 4.95 Hz,
2H); 7.25–7.26 (m, 2H); 7.47–7.51 (m, 4H); 7.59 (s, 3H);
7.89 (s, 1H); 8.53 (s, 2H); 9.88 (s, 1H); 11.45 (s, 1H). ESI–
MS: 431.1 (C22H19N6O2S, [M?H]?). Anal. Calcd for
C22H18N6O2S: C, 61.38; H, 4.21; N, 19.52. Found: C,
61.58; H, 4.22; N, 19.45.
White powder, yield 89 %; mp: 227–228 °C. 1H NMR
(300 MHz, DMSO): 4.55 (s, 2H); 7.30 (s, 2H); 7.50–7.52
(m, 4H); 7.60–7.62 (m, 3H); 7.70–7.73 (m, 2H); 8.01 (s,
1H); 8.55 (s, 2H); 11.71 (s, 1H). ESI–MS: 433.1
(C22H18FN6OS, [M?H]?). Anal. Calcd for C22H17FN6OS:
C, 61.10; H, 3.96; N, 19.43. Found: C, 61.28; H, 3.95; N,
19.47.
(E)-N0-(4-bromobenzylidene)-2-((4-phenyl-5-(pyridin-4-
yl)-4H-1,2,4-triazol-3-yl)thio)acetohydrazide (6e)
(E)-N0-(2-hydroxybenzylidene)-2-((4-phenyl-5-(pyridin-4-
yl)-4H-1,2,4-triazol-3-yl)thio)acetohydrazide (6j)
White powder, yield 89 %; mp: 232–233 °C. 1H NMR
(300 MHz, DMSO): 4.53 (s, 2H); 7.27–7.29 (m, 2H); 7.50
(s, 2H); 7.62–7.65 (m, 7H); 8.00 (s, 1H); 8.55 (d, J = 3 Hz,
2H); 11.75 (s, 1H). ESI–MS: 493.0 (C22H18BrN6OS,
[M?H]?). Anal. Calcd for C22H17BrN6OS: C, 53.56; H,
3.47; N, 17.03. Found: C, 53.41; H, 3.46; N, 17.07.
White powder, yield 88 %; mp: 242–243 °C. 1H NMR
(300 MHz, DMSO): 4.52 (s, 2H); 6.85–6.92 (m, 2H);
7.23–7.30 (m, 3H); 7.50–7.66 (m, 6H); 8.42 (s, 1H); 8.50 (s,
2H); 10.95 (s, 1H); 11.99 (s, 1H). ESI–MS: 331.1
(C22H19N6O2S, [M?H]?). Anal. Calcd for C22H18N6O2S: C,
61.38; H, 4.21; N, 19.52. Found: C, 61.57; H, 4.20; N, 19.46.
(E)-N0-(4-chlorobenzylidene)-2-((4-phenyl-5-(pyridin-
4-yl)-4H-1,2,4-triazol-3-yl)thio)acetohydrazide (6f)
(E)-N0-(5-bromo-2-hydroxybenzylidene)-2-((4-phenyl-5-
(pyridin-4-yl)-4H-1,2,4-triazol-3-yl)thio)acetohydrazide
(6j)
White powder, yield 92 %; mp: 218–219 °C. 1H NMR
(300 MHz, DMSO): 4.53 (s, 2H); 7.28 (t, J = 5.1 Hz, 2H);
7.50–7.51 (m, 4H); 7.62 (s, 3H); 7.69–7.73 (m, 2H); 8.01
(s, 1H); 8.55 (s, 2H); 11.73 (s, 1H). ESI–MS: 449.1
White powder, yield 90 %; mp: 290–291 °C. 1H NMR
(300 MHz, DMSO): 4.50 (s, 2H); 6.86–6.91 (m, 2H);
7.25–7.30 (m, 2H); 7.53–7.67 (m, 6H); 8.46 (s, 1H); 8.51 (s,
2H); 10.97 (s, 1H); 11.97 (s, 1H). ESI–MS: 509.0 (C22H18
BrN6O2S, [M?H]?). Anal. Calcd for C22H17BrN6O2S: C,
51.87; H, 3.36; N, 16.50. Found: C, 51.68; H, 3.37; N, 16.55.
(C22H18ClN6OS,
[M?H]?).
Anal.
Calcd
for
C22H17ClN6OS: C, 58.86; H, 3.82; N, 18.72. Found: C,
58.69; H, 3.83; N, 18.77.
(E)-N0-(4-fluorobenzylidene)-2-((4-phenyl-5-(pyridin-4-yl)-
4H-1,2,4-triazol-3-yl)thio)acetohydrazide (6g)
(E)-N0-(3,5-dibromo-2-hydroxybenzylidene)-2-((4-phenyl-
5-(pyridin-4-yl)-4H-1,2,4-triazol-3-yl)thio)acetohydrazide
(6k)
White powder, yield 90 %; mp: 227–228 °C. 1H NMR
(300 MHz, DMSO): 4.54 (s, 2H); 7.30 (s, 2H); 7.50–7.51 (m,
4H); 7.61–7.62 (m, 3H); 7.69–7.73 (m, 2H); 8.01 (s, 1H);
8.57 (s, 2H); 11.75 (s, 1H). ESI–MS: 433.1 (C22H18FN6OS,
[M?H]?). Anal. Calcd for C22H17FN6OS: C, 61.10; H, 3.96;
N, 19.43. Found: C, 61.29; H, 3.95; N, 19.49.
White powder, yield 88 %; mp: 270–271 °C. 1H NMR
(300 MHz, DMSO): 4.53 (s, 2H); 6.83–6.87 (m, 2H);
7.26–7.30 (m, 2H); 7.55–7.65 (m, 5H); 8.43 (s, 1H); 8.52
(s, 2H); 10.99 (s, 1H); 12.01 (s, 1H). ESI–MS: 586.9
(C22H17Br2N6O2S,
[M?H]?).
Anal.
Calcd
for
C22H16Br2N6O2S: C, 44.92; H, 2.74; N, 14.29. Found: C,
44.78; H, 2.73; N, 14.34.
(E)-N0-(2-fluorobenzylidene)-2-((4-phenyl-5-(pyridin-4-yl)-
4H-1,2,4-triazol-3-yl)thio)acetohydrazide (6h)
(E)-N0-(3,4-dihydroxybenzylidene)-2-((4-phenyl-5-
(pyridin-4-yl)-4H-1,2,4-triazol-3-yl)thio)acetohydrazide
(6m)
White powder, yield 91 %; mp: 235–236 °C. 1H NMR
(300 MHz, DMSO): 4.41 (s, 2H); 6.95–7.11 (m, 2H);
7.29–7.32 (m, 3H); 7.37–7.39 (m, 2H); 7.51 (s, 2H);
7.60–7.62 (m, 2H); 7.82 (s, 1H); 8.54 (s, 2H); 11.59 (s,
1H). ESI–MS: 433.1 (C22H18FN6OS, [M?H]?). Anal.
White powder, yield 92 %; mp: 255–256 °C. 1H NMR
(300 MHz, DMSO): 3.41–3.45 (m, 2H); 4.54 (s, 2H); 6.77
123

Med Chem Res (2013) 22:3193–3203
3203
EI-Azab AS, Al-Omar MA, Abdel-Aziz AM, Abdel-Aziz NI, El-
Sayed AA, Aleisa AM, Sayed-Ahmed MM, Abdel-Hamide SG
(2010) Design, synthesis and biological evaluation of novel
quinazoline derivatives as potential antitumor agents: molecular
docking study. Eur J Med Chem 45:4188–4198
Golubovskaya V, Kaur A, Cance W (2004) Cloning and character-
ization of the promoter region of human focal adhesion kinase
gene: nuclear factor kappaB and p53 binding sites. Biochim
Biophys Acta 1678:111–125
(d, J = 4.83 Hz, 1H); 6.89 (t, J = 5.88 Hz, 1H); 7.16 (d,
J = 10.44 Hz, 1H); 7.49–7.50 (m, 2H); 7.53–7.54 (m, 2H);
7.62–7.64 (m, 3H); 7.84 (s, 1H); 8.67–8.69 (m, 2H); 11.47
(s, 1H). ESI–MS: 447.1 (C22H19N6O3S, [M?H]?). Anal.
Calcd for C22H18N6O3S: C, 59.18; H, 4.06; N, 18.82.
Found: C, 59.01; H, 4.05; N, 18.77.
(E)-2-((4-phenyl-5-(pyridin-4-yl)-4H-1,2,4-triazol-3-
yl)thio)-N0-((E)-3-phenylallylidene)acetohydrazide (6n)
Haber J (2001) Present status and perspectives on antimycotics with
systemic effects. Cas Lek Cesk 140:596–604
Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell
100:57–70
Juan S, Yu-Shun Y, Wei L, Yan-bin Z, Xiao-Liang W, Jian-Feng T,
Hai-Liang Z (2011) Synthesis, biological evaluation and molec-
ular docking studies of 1,3,4-thiadiazole derivatives containing
1,4-benzodioxan as potential antitumor agents. Bioorg Med
Chem 21:6116–6121
Kahana O, Micksche M, Witz IP, Yron I (2002) The focal adhesion
kinase (P125FAK) is constitutively active in human malignant
melanoma. Oncogene 21:3969–3977
White powder, yield 92 %; mp: 235–236 °C. 1H NMR
(300 MHz, DMSO): 4.45 (s, 2H); 6.93–7.10 (m, 2H);
7.29–7.33 (m, 3H); 7.36–7.39 (m, 2H); 7.51 (s, 2H);
7.61–7.62 (m, 5H); 7.83 (d, J = 5.88 Hz, 1H); 8.54 (s,
2H); 11.59 (s, 1H). ESI–MS: 441.1 (C24H21N6OS,
[M?H]?). Anal. Calcd for C24H20N6OS: C, 65.44; H, 4.58;
N, 19.08. Found: C, 65.59; H, 4.57; N, 19.14.
Owens LV, Xu L, Craven RJ, Dent GA, Weiner TM, Kornberg L, Liu
ET, Cance WG (1995) Overexpression of the focal adhesion
kinase (p125FAK) in invasive human tumors. Cancer Res 55:2752
Prasad DJ, Ashok M, Karegoudar P, Poojary B, Holla BS, Suche-
taKumari N (2009) Synthesis and antimicrobial activities of
some new triazolothiadiazoles bearing 4-methylthiobenzyl moi-
ety. Eur J Med Chem 44:551–557
Acknowledgments The study was financed by National Natural
Science Foundation of China (No. J1103512) and Dr. Qiu thanks
Henan Research Program of Foundation and Advanced Technology
(112300410324, 122300410386).
Romagnoli R, Baraldi PG (2010) Synthesis and antitumor activity of
1,5-disubstituted 1,2,4-triazoles as cis-restricted combretastatin
analogues. J Med Chem 53:4248–4258
References
Schreier E, Helv (1976) Human plasma levels of some anti-migraine
drugs. Chim Acta 59:585–606
Sheldrick GM (1997) SHELX-97. Program for X-ray crystal structure
solution and refinement
Sonoda Y, Matsumoto Y, Funakoshi M, Yamamoto D, Hanks SK,
Kasahara T (2000) Anti-apoptotic role of focal adhesion kinase
(FAK). Induction of inhibitor-of-apoptosis proteins and apopto-
sis suppression by the overexpression of FAK in a human
leukemic cell line, HL-60. J Biol Chem 275:16309–16315
Stanton HLK, Robeto G, Chung HC, Marcus CWY, Eva L (2008)
Synthesis and anti-cancer activity of benzothiazole containing
phthalimide on human carcinoma cell lines. Bioorg Med Chem
16:3626–3631
Almasirad A, Tabatabai SA, Faizi M, Kebriaeezadeh A, Mehrabi N,
Dalvandi A, Shafiee A (2004) Synthesis and anticonvulsant
activity of new 2-substituted-5-[2-(2-fluorophenoxy)phenyl]-
1,3,4-oxadiazoles and 1,2,4-triazoles. Bioorg Med Chem Lett
14:6057–6059
Amir M, Shikha K (2004) Synthesis and anti-inflammatory, analgesic,
ulcerogenic and lipid peroxidation activities of some new
2-[(2,6-dichloroanilino) phenyl]acetic acid derivatives. Eur J
Med Chem 39:535–545
Aronsohn MS, Brown HM, Hauptman G, Kornberg LJ (2003)
Expression of focal adhesion kinase and phosphorylated focal
adhesion kinase in squamous cell carcinoma of the larynx.
Laryngoscope 113:1944–1948
Sztanke K, Tuzimski T (2008) Synthesis, determination of the
lipophilicity, anticancer and antimicrobial properties of some
fused 1,2,4-triazole derivatives. Eur J Med Chem 43:404–419
Van Nimwegen MJ, van de Water B (2007) Focal adhesion kinase: a
potential target in cancer therapy. Biochem Pharmacol
73:597–609
Walczak K, Gondela A, Suwinski J (2004) Synthesis and anti-
tuberculosis activity of N-aryl-C-nitroazoles. Eur J Med Chem
39:849–853
Ashok M, Holla BS (2007) Convenient synthesis of some triazolo-
thiadiazoles and triazolothiadiazines carrying 4-methylthioben-
zyl moiety as possible antimicrobial agents. J Pharmacol Toxicol
2:256–263
Bhat KS, Poojary B, Prasad DJ, Nalk B, Holla BS (2009) Synthesis
and antitumor activity studies of some new fused 1,2,4-triazole
derivatives carrying 2,4-dichloro-5-fluorophenyl moiety. Eur J
Med Chem 44:5066–5070
Budavari S, O’Neil MJ, Smith A, Heckelman PE, Kinneary JF (1996)
The Merck index, an encyclopedia of chemicals, drugs, and
biologicals, 12th edn. Merck & Co., Inc., Whitehouse Station, NJ
123