Synthesis of some new S-alkylated 1,2,4-triazoles, their Mannich bases and their biological activities

Article abstract of DOI:10.1002/ardp.201100128

A series of 1-(4-methoxyphenyl)-2-[5-{(biphenyl-4-yloxy)methyl}4- (substituted phenyl)-3-mercapto-(4H)-1,2,4-triazol-3-ylthio)] ethanones (6a-6s) and 4-(substituted phenyl)-3-(morpholin/pyrrolidin-4-ylmethylthio)-5-(4- phenylphenoxymethyl)-4H-1,2,4-triazoles (7a-7e) were synthesized in order to obtain new compounds with potent anti-inflammatory and analgesic activity with insignificant ulceration. Among the synthesized compounds, (6c), (6e), (6g) and (6l) from triazole series and (7b) and (7e) from Mannich base series were found to exhibit significant anti-inflammatory activity with 59.69, 59.69, 64.69, 79.84, 54.54, 79.69% and 52.55, 57.50, 72.52, 83.03, 60.06, 84.08% inhibition of paw edema at 3 h and 5 h respectively, in comparison to the standard drug ibuprofen (78.93 and 82.58% at 3 h and 5 h). The active compounds were further tested for their analgesic activity and gastric ulceration study. Compounds 6g, 7b and 7e exhibited significant analgesic activity with reaction time (3.60, 3.22, 3.88 s) respectively at 60 min. without causing any gastric irritation. These compounds were also screened for their in vitro antimicrobial activity, Compounds 6f, 6g, 6h, 6l, 6o, 6p, 7a, 7b and 7c showed significant zone of inhibition against various antimicrobial stains. It is concluded that the compounds 6g, 7b and 7e possess a good spectrum of activities. Compound 7e may be considered potent for development of better anti-inflammatory agent. The antimicrobial activity revealed that most of the compounds showed moderate to significant activity. Compounds containing nitro, chloro, bromo and fluoro group showing better activity. All the compounds from 7a, 7b and 7e were active against gram positive bacteria (S. aureus).

Full text of DOI:10.1002/ardp.201100128

Arch. Pharm. Chem. Life Sci. 2012, 345, 203–214
203
Full Paper
Synthesis of Some New S-Alkylated 1,2,4-Triazoles,
Their Mannich Bases and Their Biological Activities
Mohammad Mahboob Alam1, Syed Nazreen1, Saqlain Haider1, Syed Shafi1,
Mohammad Shahar Yar2, Hinna Hamid1, and Mohammad Sarwar Alam1
1 Department of Chemistry, Faculty of Science, Jamia Hamdard, New Delhi, India
2 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Jamia Hamdard, New Delhi, India
A series of 1-(4-methoxyphenyl)-2-[5-{(biphenyl-4-yloxy)methyl}4-(substituted phenyl)-3-mercapto-(4H)-
1,2,4-triazol-3-ylthio)] ethanones (6a–6s) and 4-(substituted phenyl)-3-(morpholin/pyrrolidin-4-
ylmethylthio)-5-(4-phenylphenoxymethyl)-4H-1,2,4-triazoles (7a–7e) were synthesized in order to
obtain new compounds with potent anti-inflammatory and analgesic activity with insignificant
ulceration. Among the synthesized compounds, (6c), (6e), (6g) and (6l) from triazole series and (7b)
and (7e) from Mannich base series were found to exhibit significant anti-inflammatory activity with
59.69, 59.69, 64.69, 79.84, 54.54, 79.69% and 52.55, 57.50, 72.52, 83.03, 60.06, 84.08% inhibition of paw
edema at 3 h and 5 h respectively, in comparison to the standard drug ibuprofen (78.93 and 82.58% at
3 h and 5 h). The active compounds were further tested for their analgesic activity and gastric
ulceration study. Compounds 6g, 7b and 7e exhibited significant analgesic activity with reaction
time (3.60, 3.22, 3.88 s) respectively at 60 min. without causing any gastric irritation. These
compounds were also screened for their in vitro antimicrobial activity, Compounds 6f, 6g, 6h, 6l,
6o, 6p, 7a, 7b and 7c showed significant zone of inhibition against various antimicrobial stains. It is
concluded that the compounds 6g, 7b and 7e possess a good spectrum of activities. Compound 7e may
be considered potent for development of better anti-inflammatory agent. The antimicrobial activity
revealed that most of the compounds showed moderate to significant activity. Compounds containing
nitro, chloro, bromo and fluoro group showing better activity. All the compounds from 7a, 7b and 7e
were active against gram positive bacteria (S. aureus).
Keywords: Analgesic / Antimicrobial activity / Anti-inflammatory / Mannich base / Triazole / Ulcerogenic
Received: April 6, 2011; Revised: June 7, 2011; Accepted: June 20, 2011
DOI 10.1002/ardp.201100128
Introduction
analytical reagents, agriculture products, and dyes [1]. The
development of new approaches for the synthesis of novel
heterocycles substituted with unique functional groups
forms the basis of an extensive research activity in synthetic
organic chemistry. The 1,2,4-triazole derivatives and their
Mannich bases represent an important class of heterocyclic
compounds which are known for their broad spectrum of
biological activities including antimicrobial, anti-inflamma-
tory, analgesic and many other uses [2–8]. As resistance to
anti-inflammatory and antimicrobial drugs is widespread,
there is an increasing demand for the identification of novel
structure leads that may be of use in designing new, potent
and less toxic anti-inflammatory and antimicrobial agents.
Several 1,2,4-triazole and oxadiazole based antimicrobial
agents have been synthesized to develop new molecular
Heterocycles play an important role in all spheres of life
including pharmaceuticals, natural resources, veterinary,
Correspondence: Prof. Mohammad Sarwar Alam, Department of
Chemistry, Faculty of Science, Jamia Hamdard, New Delhi, India 110 062.
E-mail: msalam@jamiahamdard.ac.in
Fax: þ91-11- 26059663
Abbreviations: NSAIDs: non-steroidal anti-inflammatory drugs; HP-
PLA2: human platelet phospholipase A2; CDCl3: deuterated
chloroform; DMSO-d6: deuterated dimethylsulfoxide; ATCC: American
Type Culture Collection; MTCC: Microbial Type Culture Collection;
AIIMS: All India Institute of Medical Science; CDRI: Central Drug
Research Institute; SDA: Sabouraud dextrose agar; MH: Muller Hinton;
CMC: carboxymethyl cellulose.
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204
M. M. Alam et al.
Arch. Pharm. Chem. Life Sci. 2012, 345, 203–214
entities with high potential against various microbial strains.
Further several non-steroidal anti-inflammatory drugs
(NSAIDs) have been developed so far to heal the inflam-
mation. There are a number of NSAIDs present in the market
to heal the inflammation but majority of them causes adverse
side effects like gastric ulcer [9], kidney damage [10] and some
of the NSAIDs also cause hepatotoxicity [11]. During the last
few decades, a considerable attention has been devoted to the
synthesis of various 1,2,4-triazole derivatives [12–16]. Several
examples of NSAIDs having triazole moieties have been cited
in the literature. Among them, 1,2,4-triazole-3-thiol deriva-
tives are of particular interest and have been studied in
recent years [17–20]. Most of the NSAIDs having biphenyl
derivatives substituted with an aromatic or heteroaromatic
aryl nucleus show better anti-inflammatory activity. Some of
the biaryldiacid derivatives have been evaluated as potential
anti-inflammatory agents through the inhibition of 14 kDa
human platelet phospholipase A2 (HP-PLA2) [21]. Some of the
biphenylic compounds that contain free carboxyl groups
cause ulceration [22–23].
phenacyl bromides/benzyl bromide leads to the formation
of target molecules (6a–6s). The triazoles (5c, 5d and 5f) were
also converted into corresponding Mannich bases (7a–7e) by
reacting with different secondary amines and formaldehyde
in presence of alcohol.
Cyclization of compounds 4(a–f) to 5(a–f) were confirmed
–
from the IR (absorption bands for S–H, 2680–2690 and C N,
–
1594–1607 cmꢀ1 triazole ring stretching) and from
the 1H NMR spectra (singlet at d 8.71–13.61 for S–H).
S-Alkylation of compounds 5(a–e) to 6(a–s) was confirmed
–
from IR (absorption bands for C O, 1655–1687) and from
13
–
C NMR spectra (peaks at d 190–192 for C O).
–
–
1,2,4-Triazole ring in compound 5(a–f) will exist in two
tautomeric forms. Under the Mannich reaction conditions
the -NH proton of the triazole ring will participate in the
reaction and will form the final Mannich base 7(a–e).
Formation of Mannich base was confirmed from 1H NMR
(presence of a singlet at d 4.47–4.93 due to methylene proton
N–CH2–N), the 13C NMR spectra of 7(a–e) displayed peaks in
the range of d 68.76–68.82 for the carbon bonded with two
nitrogen (N–CH2–N) and the peaks in the range of d 170–172
for the carbon in the triazole ring attached with sulphur
In view of the importance of biaryl and 1,2,4-triazole moi-
eties as anti-inflammatory and antimicrobial agents, we aim
to develop some novel biologically interesting moieties
encompassing biaryl and 1,2,4-triazoles moieties conjugated
through an ether linkage. A series of novel S-substituted 1,2,4-
triazoles and Mannich bases of 1,2,4-triazoles were synthes-
ized starting from p-hydroxy biphenyl. The structures of new
compounds were confirmed by chemical and spectroscopic
methods like FT-IR, 1H NMR, 13C NMR spectroscopy and mass
spectrometry. All the synthesized compounds were screened
for their biological activities viz. anti-inflammatory, analge-
sic, ulcerogenic and antimicrobial activity. The synthetic
path of the target compounds is shown in Scheme 1.
–
(C S). All these peaks were absent in the compounds (5a–5e)
–
and the spectral data are in line with Barbuceanu et al. [25]
which confirms the formation of the compounds (7a–7e).
Anti-inflammatory activity
The results of anti-inflammatory activity are summarized in
Table 1. Among the tested compounds, 6c, 6e, 6g and 6l from
triazole series and 7b, 7e from Mannich base series were
found to be active and have significant activity (59.69,
59.69, 64.69, 79.84, 54.54, 79.69% and 52.55, 57.50, 72.52,
83.03, 60.06, 84.08% inhibition in paw oedema at 3 h and 5 h
respectively) when compared to the standard drug ibuprofen
(78.93 and 82.58% at 3 h and 5 h). Compounds 6l and 7e were
exhibited more active than the standard drug ibuprofen.
From the biological data, the structure activity relation-
ship (SAR) can be drawn as follows. Compounds having no
substitution on the aromatic ring attached to the 1,2,4-tri-
azollyl ring and the electron withdrawing group on the
phenacyl aromatic ring are showing better activity.
Substitution of electron donating groups on triazolyl
attached aromatic ring decreases the activity but is compar-
able with the standard. Whereas when the same aromatic
ring was substituted with electron withdrawing groups the
activity significantly decreases. Compounds having bulkier
substitutions like -Br and –NO2 groups on the phenacyl aro-
matic ring are showing potential activity. Whereas the
simple benzylation on the 3-mercapto 1,2,4-triazoles signifi-
cantly diminishes the activity. These data suggests that the
carbonyl functionality of the phenacyl group is playing an
important role in the enzymatic interactions responsible for
Results and discussion
Chemistry
Twenty-four new compounds (6a–6s) and (7a–7e) were syn-
thesized starting from p-hydroxybiphenyl as outlined in
Scheme 1. Reaction of p-hydroxybiphenyl (1), with ethyl-
chloroacetate in presence of potassium carbonate in anhy-
drous acetone afforded ethyl 2-(biphenyl-4-yloxy) acetate (2)
which readily yielded 2-(biphenyl-4-yloxy) acetohydrazide (3)
by refluxing with hydrazine monohydrate in absolute alco-
hol. Compound (3) then was converted into corresponding
thiosemicarbazide (4a–4f) by reacting with different substi-
tuted aryl isothiocyanate in absolute alcohol using the
known protocol [24]. The thiosemicarbazides (4a–4f) were
further converted into respective 5-[(biphenyl-4-yloxy)-
methyl]4-(substituted phenyl)-3-mercapto-(4H)-1,2,4-triazoles
(5a–5f) using triethyl amine in absolute alcohol. Finally
S-alkylation of 3-mercapto-1,2,4-triazoles with substituted
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Arch. Pharm. Chem. Life Sci. 2012, 345, 203–214
1,2,4-Triazole Derivatives as Potent Bio-Active Agents
205
a
OH
O
CH2COOC2H5
1
2
b
O
3
CH2CONHNH2
c
4a. R1=4 Cl
4b. R1=4 NO2
4c. R1=4 OMe
4d. R1= H
H
N
O
CH2CONHNH
4a-4f
4e. R1= F
S
4f R= 2Me
R1
NH
N
d
S
N
N
O
N
SH
O
N
5a-5f
R1
R1
f
e
R3
S
N
N
N
N
S
R2
O
N
O
N
6a-6s
7a-7e
R1
R1
R2
R3
R1
Comp.
7a
Comp.
6a
R2
Comp.
R1
R1
O
O
O
H3C
4-Cl
6b
6d
6f
4-Cl
N
O
N
4-Cl
O
O
O
6c
6e
4-Cl
4-Cl
4-OMe
4-OMe
O2N
7b
7c
Br
N
CH3
O
4-NO2
4-NO2
N
O
O
O
H
6g
Cl
7d
7e
4-OMe
N
N
O
O
Br
Br
6h
6j
4-OMe
4-OMe
O
O
O
6i
2-Me
4-OMe
H
H3C
O
6k
6m
O2N
6l
H
H
6n
O2N
Br
H
O
O
6o
4-F
4-F
6p
6r
O
O
6q
6s
4-Cl
4-F
4-F
H3C
O2N
Reagents and conditions: (a) Dry acetone, anhydrous K2CO3, ethylchloroacetate, reflux 10 h; (b) NH2NH2.H2O, Abs.
alcohol, reflux 3 h; (c) Abs. alcohol, ArNCS, reflux 4−6 h; (d) Abs. alcohol, TEA, reflux 6−8 h (e) Abs. alcohol, Na metal,
substituted phenacylbromide, reflux 4−6 h (f) Abs. alcohol, HCHO, amines, stirring at room temperature, 24 h.
Scheme 1. Synthetic route employed for the synthesis of target molecules.
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206
M. M. Alam et al.
Arch. Pharm. Chem. Life Sci. 2012, 345, 203–214
Table 1. Results of anti-inflammatory activity of azole derivatives against carrageenan induced rat paw edema model.
Compounds
Change in paw edema volume (mL)
After drug treatment
Antiinflamatory activity
% Inhibition
3 h
5 h
3 h
5 h
6a
6b
6c
6d
6e
6f
6g
6h
6i
6j
6k
6l
6m
6n
6o
6p
6q
6r
6s
7a
7b
7c
7d
0.319 ꢁ 0.067ꢂꢂꢂ
0.341 ꢁ 0.056ꢂꢂꢂ
0.266 ꢁ 0.071ꢂꢂꢂ
0.566 ꢁ 0.049ns
0.266 ꢁ 0.042ꢂꢂꢂ
0.583 ꢁ 0.065 ns
0.233 ꢁ 0.049ꢂꢂꢂ
0.423 ꢁ 0.058ꢂꢂ
0.533 ꢁ 0.042 ns
0.330 ꢁ 0.049ꢂꢂꢂ
0.300 ꢁ 0.036ꢂꢂꢂ
0.133 ꢁ 0.021ꢂꢂꢂ
0.583 ꢁ 0.060ꢂns
0.466 ꢁ 0.061ꢂꢂꢂ
0.300 ꢁ 0.051ꢂꢂꢂ
0.298 ꢁ 0.057ꢂꢂꢂ
0.295 ꢁ 0.046ꢂꢂ
0.345 ꢁ 0.036
0.321 ꢁ 0.075ꢂꢂꢂ
0.338 ꢁ 0.068ꢂꢂꢂ
0.316 ꢁ 0.090ꢂꢂꢂ
0.450 ꢁ 0.125 ns
0.283 ꢁ 0.040ꢂꢂꢂ
0.600 ꢁ 0.089 ns
0.183 ꢁ 0.040ꢂꢂꢂ
0.413 ꢁ 0.057 ns
0.483 ꢁ 0.118 ns
0.366 ꢁ 0.055ꢂꢂ
0.283 ꢁ 0.040ꢂꢂꢂ
0.113 ꢁ 0.033ꢂꢂꢂ
0.383 ꢁ 0.047ꢂ
0.466 ꢁ 0.033 ns
0.366 ꢁ 0.076ꢂꢂ
0.300 ꢁ 0.057ꢂꢂꢂ
0.279 ꢁ 0.036ꢂꢂꢂ
0.331 ꢁ 0.039ꢂꢂ
0.233 ꢁ 0.091ꢂꢂꢂ
0.367 ꢁ 0.047ꢂꢂ
0.266 ꢁ 0.088ꢂꢂꢂ
0.436 ꢁ 0.074 ns
0.516 ꢁ 0.119 ns
0.106 ꢁ 0.047ꢂꢂꢂ
0.116 ꢁ 0.051ꢂꢂꢂ
0.666 ꢁ 0.088
51.66
48.33
59.69
14.24
59.69
11.66
64.69
35.90
19.24
50.00
54.54
79.84
11.66
28.64
54.54
54.69
55.30
47.72
46.96
41.36
54.54
37.87
14.24
79.69
78.93
–
51.36
49.24
52.55
32.43
57.50
09.00
72.52
37.98
27.24
45.04
57.50
83.03
42.49
29.39
45.04
54.95
58.10
50.30
65.01
44.89
60.06
34.53
22.52
84.08
82.58
–
0.350 ꢁ 0.042ꢂꢂꢂ
0.387 ꢁ 0.043ꢂꢂ
0.300 ꢁ 0.066ꢂꢂꢂ
0.410 ꢁ 0.072ꢂꢂꢂ
0.566 ꢁ 0.042ns
0.134 ꢁ 0.055ꢂꢂꢂ
0.139 ꢁ 0.021ꢂꢂꢂ
0.660 ꢁ 0.042
7e
Ibuprofen
Control
Data analyzed by one-way ANOVA followed by Dunnett’s ‘t’ test (n ¼ 6).
ꢂ
ꢂꢂ
ꢂꢂꢂ
p <0.05, p <0.01 & p <0.001 significantly different from standard; ns, not significant.
anti-inflammatory activity. As aryl substituted 1,2,4-triazoles
and some of the other azole derivatives (celocoxib) are
reported to be selective inhibitors of COX-2 [26], the possible
mode of action of above synthesized compounds might be
similar to other azole drugs. Overall the compound 6l sub-
stituted with nitro group at the para position on the aromatic
ring of the phenacyl group is showing significant activity
compared to standard drug ibuprofen.
From the Mannich base series, compounds having morpho-
line ring and the substitution of electron donating groups on
Table 2. Results of analgesic activity of the synthesized compounds.
Compounds
Mean value of tail flick latency (s) W SEM
0 min
30 min
60 min
120 min
6c
6e
6l
6g
7b
7e
1.93 ꢁ 0.33
2.07 ꢁ 0.368
2.16 ꢁ 0.298
1.88 ꢁ 0.324
2.08 ꢁ 0.32
1.93 ꢁ 0.303
2.03 ꢁ 0.330
1.98 ꢁ 0.307
3.04 ꢁ 0.284
3.81 ꢁ 0.257
2.85 ꢁ 0.367
3.23 ꢁ 0.370
3.80 ꢁ 0.233
3.86 ꢁ 0.346
1.94 ꢁ 0.236
3.716 ꢁ 0.388
2.84 ꢁ 0.302
3.96 ꢁ 0.377
3.91 ꢁ 0.529
3.603 ꢁ 0.341
3.225 ꢁ 0.264
3.88 ꢁ 0.243
2.02 ꢁ 0.192
3.956 ꢁ 0.473
2.85 ꢁ 0.372
3.08 ꢁ 0.499
2.516 ꢁ 0.323
2.56 ꢁ 0.227
2.94 ꢁ 0.232
2.85 ꢁ 0.237
2.85 ꢁ 0.237
2.76 ꢁ 0.212
Control
Ibuprofen
Data analyzed by one-way ANOVA followed by Dunnett’s ‘t’ test (n ¼ 6), ꢂp <0.05, ꢂꢂp <0.01 & ꢂꢂꢂp <0.001 significantly different from
standard; ns, not significant.
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Arch. Pharm. Chem. Life Sci. 2012, 345, 203–214
1,2,4-Triazole Derivatives as Potent Bio-Active Agents
207
the aromatic ring attached to triazollyl ring are showing
better activity. Compound 7e with methyl substitution at
the ortho position and with morpholine ring is showing
significant activity compared to the standard ibuprofen.
All these data were analyzed by one-way ANOVA test
followed by Dunnet’s test in carrageenan induced rat paw
oedema model.
phenacyl group are showing better activity than the simple
benzylated ones.
Compounds 6l, 6p and 6q showed moderate activity
against bacterial strain Pseudomonas aeruginosa whereas com-
pounds 7a, 7b and 7e from the Mannich base series showed
good activity against Staphylococcus aureus (Gram possitive
bacteria). By closely observing the results it can be concluded
that most of the halogen substituted compounds (6g, 6h, 6o,
6q, 6p) displayed good activity against various fungal strains.
Gastric ulceration study
When compared with ibuprofen, compounds 6g, 7b and 7e
did not cause any gastric ulceration and disruption of gastric Experimental
epithelial cells at 3 times the dose used for anti-inflammatory
activity (Experimental). Hence gastric tolerance to these com-
pounds was better than that of ibuprofen indicating that
carboxylic group present in the ibuprofen is responsible for
ulceration [27]. Stomach wall of ibuprofen treated group
at low power (10x) photomicrograph showed damage of
the mucosa and the submucosa. Stomach wall of the same
section at high power (40x) photomicrograph showed
desquamated epithelial cells in the lumen whereas in tested
compounds treated (6c), (6e) and (6l) animals, surface
epithelial damage was significant, slight submucosal damage
was seen, however there was lesser damage in comparison to
the ibuprofen. Stomach wall of 6g, 7b and 7e treated animal
showed no damage of any layer. The results are shown in
Table 3 and Figure 1.
Chemistry
Melting points were determined on an Electro thermal appar-
atus and are uncorrected. 1H NMR spectra were recorded on a
Bruker spectrometer (300, 400 MHz), chemical shifts are
reported in parts per million (ppm) relative to tetramethylsilane
(TMS) as an internal standard. Coupling constants J (H–H) are in
Hertz (Hz). FT-IR spectra were recorded on a Bruker spectrometer,
Mass spectra were recorded on a Jeol JMS-D 300 instrument fitted
with a JMS 2000 data system at 70 eV. Elemental analyses were
recorded on CHN analyzer model and found within range
(ꢁ0.4%) of theoretical value.
General procedure for the synthesis of ethyl-2-biphenyl-4-
yloxy-acetate (2)
To a mixture of p-hydroxybiphenyl (10 mM) and ethylchloroace-
tate (10 mM) in 50 mL of anhydrous acetone was added K2CO3
(36 mM). The suspension was refluxed for 20 h, after completion
of reaction, monitored by TLC, the reaction mixture was filtered
in hot condition, the filtrate was concentrated and finally the
crude product was crystallized from methanol in cold condition.
Yield 75%; white flakes; m.p. 40–428C; IR (KBr) cmꢀ1: 3055
Antimicrobial activity
All the newly synthesized compounds were tested against
various microbial strains for their anti-microbial activity
(Table 4). Some of the compounds from both the series have
shown moderate to good activity against certain strains.
Compound 6f showed moderate activity against all the tested
fungal strains whereas compounds 6g and 6h were active
against Aspergillus flavus. Compounds 6g, 6j, 6o, 6q and 7d
showed moderate activity against Candida albicans. From the
results it can be depicted that the compounds containing
(Ar-CH), 2921 (CH2), 1674 (C O), 1108 (C–O); 1H NMR
–
–
(300 MHz, CDCl3) d 7.54–7.53 (m, 4H), 7.42–7.36 (d, J ¼ 7.6 Hz,
2H), 7.31–7.24 (m, 1H, Ar–H), 7.04–7.01 (d, J ¼ 8.6 Hz, 2H), 5.29
(s, 2H, O–CH2), 3.92 (q, J ¼ 5.8 Hz, 2H), 1.85 (t, J ¼ 5.6 Hz, 3H);
FAB-MS (m/z): 256 (Mþ), 257 (Mþ1); Anal. Calcd. for C16H16O3:
C, 74.98; H, 6.29. Found C, 74.68; H, 6.21.
General procedure for synthesis of hydrazide (3)
The ethanolic solution of ethyl-2-biphenyloxy-acetate (1) (10 mM)
and of hydrazine monohydrate (10 mM) was refluxed for 4 h.
After the completion of reaction, the reaction mixture was
cooled and the white flakes so obtained were filtered and recrys-
tallized from alcohol.
Table 3. Results of gastric ulcerogenic study.
Group
Surface
epithelial
damage
Submucosal
damage
Deep
mucosal
damage
Muscular
layer
damage
Yield 90%; white flakes; m.p. 165–1678C; IR (KBr) cmꢀ1: 3350
1
(N–H), 3055 (Ar–H), 2921 (CH ), 1674 (C O), 1108 (C–O); H NMR
(300 MHz, CDCl3)
–
–
d 7.52–7.51 (m, 4 H, Ar–H), 7.43–7.37
2
6c
6e
6l
6g
7b
7e
þþ
þþ
þþ
ꢀ
þþ
þþ
þþ
ꢀ
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
–
(d, J ¼ 7.8 Hz, 2H), 7.32–7.25 (m, 1H, Ar–H), 7.05–7.02
(d, J ¼ 8.5 Hz, 2H), 6.29 (s, 1H, NH), 5.21 (s, 2H, O–CH2), 3.2
(s, 2 H, NH2); FAB-MS (m/z): 242 (Mþ), 243 (Mþ1); Anal. Calcd.
for C14H14N2O2: C, 69.41; H, 5.82; N, 11.56. Found: C, 69.13; H,
5.84; N, 11.51%.
ꢀ
ꢀ
ꢀ
ꢀ
Control
Ibuprofen
ꢀ
ꢀ
General procedure for synthesis of thiosemicarbazides
(4a–4f)
þþþ
þþþ
To a solution of hydrazide (3) (1 mM) in absolute alcohol (50 mL)
was added arylisothiocyanate (1 mM) and the reaction mixture
[ꢀ], No damage; [þ], [þþ], [þþþ], indicates increasing degree of
damage
ß 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

208
M. M. Alam et al.
Arch. Pharm. Chem. Life Sci. 2012, 345, 203–214
Table 4. Results of antimicrobial activity.
Compounds
Anti-fungal activity zone of inhibition (mm) 200 (100 mg)
Anti-bacterial activity zone of inhibition (mm) 200 (100 mg)
A.f
A.n
C.a
C.c
K.p
P.a
E.c
S.a
6a
19 ꢁ 1.6 (8 ꢁ .6)
12 ꢁ 1.4 (6 ꢁ .4)
12 ꢁ 1.6 (6 ꢁ .4)
14 ꢁ 1.2 (8 ꢁ 1)
10 ꢁ 1.3 (8 ꢁ 1.6)
14 ꢁ 1.4 (2 ꢁ .7) 8 ꢁ .5 (6 ꢁ .3)
10 ꢁ 1.5 (6 ꢁ .5) 8 ꢁ1.6 (6 ꢁ .6)
6 ꢁ .3 (ꢀ)
–
6 ꢁ .3 (ꢀ)
–
10 ꢁ 1.2 (6 ꢁ .5)
8 ꢁ .8 (ꢀ)
9 ꢁ .8 (ꢀ)
8 ꢁ .5 (ꢀ)
10 ꢁ 1.1 (8 ꢁ .5)
8 ꢁ .8 (ꢀ))
10 ꢁ 1.1 (ꢀ)
–
–
6b
–
6c
10 ꢁ 1.4 (6 ꢁ .3)
12 ꢁ 1.4 (8 ꢁ .4)
12 ꢁ 1.6 (10)
8 ꢁ 3 (6 ꢁ .4)
10 ꢁ 1.4 (6)
8 ꢁ .9 (ꢀ)
6 ꢁ .4 (ꢀ)
8 ꢁ .5 (ꢀ)
12 ꢁ 1.3 (6)
12 ꢁ 1.2 (6 ꢁ .4)
9 ꢁ .8 (ꢀ)
8 ꢁ .4 (ꢀ)
6d
8 ꢁ .6 (6 ꢁ 1.6)
8 ꢁ 1.2 (6 ꢁ .7)
10 W.8 (8 W .6)
10 W .8 (6 W .4)
6 ꢁ .5 (ꢀ)
10 ꢁ .9 (8 ꢁ .7)
14 ꢁ 1.6(10 ꢁ .6)
10 ꢁ 1.2 (6 ꢁ .4)
10 ꢁ 1.6 (6 ꢁ .6)
11 ꢁ1.2 (6 ꢁ .4)
12 ꢁ 1.1 (6 ꢁ .6)
14 ꢁ 1.6 (10 ꢁ 1.2)
14 ꢁ 1.2 (6 ꢁ .6)
8 ꢁ.5 (6 ꢁ .4)
12 ꢁ1.6 (8 ꢁ .6)
10 ꢁ 1.2 (6 ꢁ .6)
12 ꢁ1.2 (8 ꢁ .7)
8 ꢁ .6 (6 ꢁ .3)
8 ꢁ.6 (6 ꢁ .4)
8 ꢁ .6 (6 ꢁ .4)
12 ꢁ 1.2 (8 ꢁ .6)
8 ꢁ .4 (ꢀ)
10 ꢁ 1.2 (8 ꢁ .6)
6e
–
6f
22 W 1.8 (12 W 1.6) 18 W .8 (10 W .4) 12 W 1.1(8 W .5)
6 ꢁ .5 (ꢀ)
–
6g
22 W1.9 (10 W 1.2)
20 W 1.4 (10 W 1.2)
18 ꢁ1.2 (8 ꢁ 1.6)
14 ꢁ .6 (7 ꢁ .5) 10 ꢁ 1.1(6 ꢁ .4)
8 ꢁ 1 (ꢀ)
–
–
6h
12 ꢁ.9 (6 ꢁ .5)
10 ꢁ.8 (8 ꢁ .7)
8 ꢁ.8 (6 ꢁ .4)
10 ꢁ.9 (6 ꢁ .4)
8 ꢁ.8 (6 ꢁ .5)
8 ꢁ.6 (6 ꢁ .4)
6 ꢁ 1.6 (5 ꢁ .4)
8 ꢁ .9 (6 ꢁ .5)
6 ꢁ .5 (ꢀ)
9 ꢁ 1 (ꢀ)
–
–
6i
8 W 9 (6 ꢁ 4)
10 W.9 (6 W 5)
8 ꢁ .8 (6 ꢁ .6)
8 ꢁ .6 (ꢀ)
–
10 ꢁ 1.2 (ꢀ)
10 ꢁ 1.2 (5 ꢁ .4)
8 ꢁ .7 (6 ꢁ .6)
–
–
6j
18 ꢁ 1.8 (10 ꢁ 1.1) 12 ꢁ 1.2 (6 ꢁ .4)
11 ꢁ .9 (ꢀ)
12 ꢁ 1.2 (8 ꢁ .6)
6k
18 ꢁ 1.5 (10 ꢁ .8)
18 ꢁ 1.3 (10 ꢁ 1.2)
18 ꢁ1.6 (8 ꢁ .6)
6 ꢁ .5 (–)
10 ꢁ1 (6 ꢁ .5)
10 ꢁ1.2(6 ꢁ .4)
8 ꢁ.6 (ꢀ)
–
6l
14 W 1.7 (10 W 1) 10 ꢁ 1.2 (6 ꢁ .6)
8 ꢁ.7 (ꢀ)
8 ꢁ.6 (ꢀ)
–
6m
8 ꢁ.9 (6 ꢁ .6)
9 ꢁ 1.2 (6 ꢁ .8)
10 W .6 (6 W .4)
8 ꢁ 1.2 (6 ꢁ .4)
10 W.8 (8 W .6)
8 ꢁ .8 (6 ꢁ .4)
9 W.8 (7 W .6)
9 ꢁ.6 (6 ꢁ .4)
–
12 ꢁ1.2 (8 ꢁ .7)
10 ꢁ .9 (6 ꢁ .6)
6n
14 ꢁ1.6 (10 ꢁ .9)
8 ꢁ .6(ꢀ)
–
60
18 ꢁ 1.8 (10 ꢁ 1.2) 12 ꢁ 9 (6 ꢁ .4)
6 ꢁ.6 (ꢀ)
12 ꢁ 1.2 (9 ꢁ .6)
10 ꢁ 1.2 (ꢀ)
8 ꢁ 1.2 (6 ꢁ .6)
6 ꢁ .6 (ꢀ)
6 ꢁ.4 (ꢀ)
6 ꢁ .3 (ꢀ)
–
6p
14 ꢁ1.2 (8 ꢁ .7)
12 ꢁ1.2 (6 ꢁ .4)
8 ꢁ.6 (ꢀ)
12 ꢁ 1.1 (8 ꢁ .4) 8 ꢁ .8 (6 ꢁ .3)
14 W 1.2 (10 W .8)
8 ꢁ .6 (ꢀ)
6q
10 ꢁ 1.2 (8 ꢁ .6)
10 ꢁ .6 (6 ꢁ .4)
10 ꢁ 1.2 (6 ꢁ .6)
8 ꢁ .6 (ꢀ)
8 ꢁ 6 (ꢀ)
8 ꢁ 7 (ꢀ)
14 W 1.2 (10 W .7)
8 ꢁ .6 (ꢀ)
6r
10 ꢁ .8 (6 ꢁ .6)
6 ꢁ .3 (ꢀ)
6s
14 ꢁ1.2 (6 ꢁ .3)
8 ꢁ.6 (6 ꢁ .4)
6 ꢁ.6 (ꢀ)
8 ꢁ 6 (5 ꢁ .3)
6 ꢁ .4 (ꢀ)
–
–
–
–
–
7a
–
12 W 1.2 (6 W .3)
12 W 1.2 (6 W .6)
10 ꢁ 8 (8 ꢁ .7)
7b
8 ꢁ 7 (ꢀ)
10 ꢁ .8 (8 ꢁ .6)
10 ꢁ 1.2(8 ꢁ .5)
9 ꢁ .4 (ꢀ)
–
8 ꢁ .6 (ꢀ)
8 ꢁ 1.6 (4)
8 ꢁ .4 (ꢀ)
Nt
7c
6 ꢁ .4 (ꢀ)
8 ꢁ 8 (ꢀ)
8 ꢁ 1.2 (8 ꢁ .6)
10 ꢁ.9 (6 ꢁ .4)
7d
12 ꢁ 1.6 (6 ꢁ .4)
10 ꢁ 1.2 (6 ꢁ .4)
10 ꢁ 1.6 ꢁ1.6)
11 ꢁ 1 (6 ꢁ .3)
10 ꢁ 1.6(6 ꢁ .8) 1 0 W 1.6(8 W 1.2) 10 ꢁ 1.6 (5 ꢁ 1.6)
8 ꢁ 1.6 (6 ꢁ 1.6) 10 ꢁ 1.6 (6 ꢁ 1.6)
7e
8 ꢁ .6 (ꢀ)
8 ꢁ .6 (6 ꢁ .6)
10 ꢁ 1.2 (6 ꢁ .4)
Nt
10 ꢁ 1.2 (6 ꢁ .6)
Nt
12 W .9 (6 W .5)
FLuconazole
Amoxicillin
Control
22 ꢁ 1.8 (12 ꢁ .9) 16 ꢁ 1 (10 ꢁ .6) 14 ꢁ 1.2(10 ꢁ .) 10 W 1.2 (8 W .7)
nt
Nt
nt
nt
nt
16 ꢁ 1 (12 ꢁ .8)
14 ꢁ .8 (10 ꢁ .5)
–
12 ꢁ .7 (8 ꢁ .4)
12 ꢁ .6 (6 ꢁ .3)
–
–
–
–
–
–
–
A.n.: Aspergillus niger [MTCC 8189], A.f.: Aspergillus flavus [MTTC 277], K.p.: Klebsiella pneumoniae [ATCC 700603] P.a.: Pseudomonas
aeruginosa [ATCC-27853], Escherichia coli [ATCC 25922], Staphylococcus aureus [ATCC 25923], DMSO [dimethylsulfoxide], Flu.: fluconazole,
Amp.: ampicillin, [nt] not tested, [–] no zone of inhibition.
was refluxed for 4–6 h. After the completion of reaction, moni-
tored by TLC, the reaction mixture was cooled to room tempera-
ture and the white crystals so formed were filtered and
crystallized from alcohol to yield the pure hydrazide (4a–4g).
1-[2-(4-Biphenyloxy)acetyl]-4(4-chlorophenyl)thiosemicarbazide (4a) Yield
74%; white crystalss; m.p. 183–1858C; IR (KBr) cmꢀ1: 3322 (N–H),
(300 MHz, DMSO-d6) d 9.21 (brs, 1H, CONH), 9.12 (brs, 1H,
CSNH), 8.92 (brs, 1H, ArNH), 7.61–7.47 (m, 6H), 7.46
(t, J ¼ 5.7 Hz, 2H), 7.33 (t, J ¼ 7.5 Hz, 1H), 7.08–7.01 (m, 4H),
4.68 (s, 2H, O–CH2), 3.88 (s, 3H, O–CH3); FAB-MS (m/z): 407 (Mþ),
408 (Mþ1); Anal. Calcd. for C22H21N3O3S: C, 64.85; H, 5.19;
N, 10.31. Found: C, 65.08; H, 5.22; N, 10.26.
2958 (Ar–H), 1714 (C O), 1195 (C S), 1108 (C–O); 1H NMR
1-[2-(4-Biphenyloxy)acetyl]-4-phenylthiosemicarbazide (4d) Yield 77%;
–
–
–
–
(300 MHz, DMSO-d6) d 10.34 (brs, 1H, CONH), 9.70 (brs, 1H,
CSNH), 9.21 (brs, 1H, ArNH), 7.52–7.47 (m, 6H), 7.43–7.36
(m, 4H), 7.30 (t, J ¼ 5.7 Hz, 1H), 6.94 (d, J ¼ 6.4 Hz, 2H), 4.69
(s, 2H, O–CH2); FAB-MS (m/z): 411 (Mþ), 412 (Mþ1); Anal. Calcd.
for C21H18ClN3O2S: C, 61.23; H, 4.40; N, 10.20. Found: C, 61.41;
H, 4.48; N, 10.18%.
white crystals; m.p. 138–1408C; IR (KBr) cmꢀ1: 3324 (N–H), 2945
(CH ), 1701 (C O), 1212 (C S), 1109 (C–O); 1H NMR (300 MHz,
–
–
–
–
2
DMSO-d6) d 10.14 (brs, 1H, CONH), 9.21 (brs, 1H, CSNH), 8.92
(brs, 1H, ArNH), 7.54–7.44 (m, 4H), 7.42–7.25 (m, 8H), 7.03
(d, J ¼ 8.6 Hz, 2H), 4.68 (s, 2H, O–CH2); FAB-MS (m/z): 377 (Mþ),
378 (Mþ1); Anal. Calcd. for C21H19N3O2S: C, 66.82; H, 5.07;
N, 11.13. Found: C, 66.48; H, 5.15; N 11.25.
1-[2-(4-Biphenyloxy)acetyl]-4(4-nitrophenyl)thiosemicarbazide (4b) Yield
69%; light yellow crystalss; m.p. 198–2008C; IR (KBr) cmꢀ1: 3246
1-[2-(4-Biphenyloxy)acetyl]-4(4-fluorophenyl)thiosemicarbazide (4e) Yield
1
(N–H), 2958 (CH ), 1693 (C O), 1208 (C S), 1113 (C–O); H NMR
76%; white crystals; m.p. 186–1888C; IR (KBr) cmꢀ1: 3244 (N–H),
–
–
–
–
2
– –
3070 (Ar–H), 2950 (CH ), 1683 (C O), 1202 (C S), 1106 (C–O);
– –
2
(300 MHz, DMSO-d6): d 9.58 (brs, 1H, CONH), 8.81 (brs, 1H, CSNH),
8.02 (brs, 1H, ArNH), 8.38 (d, J ¼ 8.6 Hz, 2H), 7.94 (d, J ¼ 8.6 Hz,
2H), 7.52–7.26 (m, 7H), 6.94 (d, J ¼ 6.6 Hz, 2H), 4.69 (s, 2H,
O–CH2). FAB-MS (m/z): 422 (Mþ), 423 (Mþ1); Anal. Calcd.
for C21H18N4O4S: C, 59.70; H, 4.29; N, 13.26. Found: C, 59.92;
H, 4.32; N, 13.20.
1H NMR (300 MHz, DMSO-d6) d 9.56 (brs, 1H, CONH), 8.96 (brs,
1H, CSNH), 8.67 (brs, 1H, ArNH), 7.88 (d, J ¼ 6.4 Hz, 2H), 7.51–7.46
(m, 4H), 7.42–7.37 (m, 4H), 7.31–7.19 (m, 3H), 4.66 (s, 2H, O–CH2);
FAB-MS (m/z): 395 (Mþ), 396 (Mþ1); Anal. Calcd. for C21H18FN3O2S: C,
63.78; H, 4.59; N, 10.63. Found: C, 63.98; H, 4.53; N, 10.58.
1-[2-(4-Biphenyloxy)acetyl]-4(2-methylphenyl)thiosemicarbazide (4f) Yield
71%; white crystals; m.p. 138–1408C; IR (KBr) cmꢀ1: 3320 (N–H),
1-[2-(4-Biphenyloxy)acetyl]-4(4-methoxyphenyl)thiosemicarbazide (4c)
Yield 72%; white crystalss; m.p. 176–1788C; IR (KBr) cmꢀ1: 3246
1
(N–H), 2953 (CH ), 1702 (C O), 1205 (C S), 1109 (C–O); H NMR
–
–
–
–
–
–
–
–
2948 (CH ), 1704 (C O), 1215 (C S), 1254 (C C), 1204 (C C), 1102
–
–
–
–
2
2
ß 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2012, 345, 203–214
1,2,4-Triazole Derivatives as Potent Bio-Active Agents
209
(C–O); 1H NMR (200 MHz, DMSO-d6) d 10.12 (brs, 1H, CONH), 9.21
(brs, 1H, CSNH), 8.92 (brs, 1H, ArNH), 7.56–7.30 (m, 11H), 6.86 (2H,
d, J ¼ 6.4 Hz), 4.67 (s, 2H, O–CH2), 2.38 (s, 3 H, CH3); Maldi-MS ( m/z):
391 (Mþ), 392 (Mþ1); Anal. Calcd. for C22H21N3O2S: C, 67.50; H,
5.07; N, 10.73. Found: C, 67.85; H, 5.11; N 10.68.
7.54–7.51 (m, 4H), 7.43 (m, 7H), 6.89 (2H, d, J ¼ 6.6 Hz), 4.68
(s, 2H, O–CH2), 2.54 (3H, s, CH3); Maldi-MS (m/z): 373 (Mþ), 374
(Mþ1). Anal. Calcd. for C22H19N3OS: C 70.75; H 5.13; N 11.25.
Found: C 70.82; H 5.10; N 11.30%.
General method for synthesis of 1(aryl)-2-[5-{(biphenyl-4-
yloxy)methyl}4-aryl-3-mercapto-(4H)-1,2,4-triazol-3-
ylthio)] ethanone/ethane (6a–6s)
General method for synthesis of 5-[(biphenyl-4-yloxy)-
methyl]4-aryl-3-mercapto-(4H)-1,2,4-triazole (5a–5f)
To a solution of thiosemicarbazide (4a–4f) (5 mM) in absolute
alcohol (50 mL) was added triethyl amine (1 mL) and the reaction
mixture was refluxed for 4–8 h. After the completion of reaction,
monitored by TLC, the reaction mixture was cooled to room
temperature and concentrated under reduced pressure
(25 mL), the concentrated solution was poured on crushed ice,
the precipitate obtained was filtered off and washed with
cold water, dried and crystallized in ethanol to yield the pure
3-mercapto-1,2,4-triazoles (5a–5f).
To a solution of 3-mercapto-1,2,4-triazole (5a–5e, 0.5 mM) in
absolute ethanol (50 mL) was added sodium metal (0.5 mM)
and different substituted phenacyl bromide (0.5 mM) and the
reaction mixture was refluxed for 1–6 h; after completion of
reaction (monitored by TLC), the reaction mixture was concen-
trated under reduced pressure (25 mL), the concentrated
solution was cooled and the crystals obtained were filtered,
washed with water, dried and recrystallized from dichlorome-
thane (DCM) and methanol to yield the pure compound (6a–6s).
2-{5-[(4-Biphenyloxy)methyl]-4-(4-chlorophenyl)-4H-1,2,4-triazol-3-ylthio}-1-
(phenyl)ethanone (6a) Yield 68%; white crystals; m.p. 192–1948C; IR
5-[(Biphenyl-4-yloxy)methyl]4-(4-chlorophenyl)-3-mercapto-(4H)-1,2,4-triazole
(5a) Yield 85%; white crystals; m.p. 228–2308C; IR (KBr) cmꢀ1: 3056
(Ar–H), 2922 (CH ), 2690 (S–H), 1606 (C N); 1H NMR (300 MHz,
–
(KBr) cmꢀ1: 2924 (CH ), 1680 (C O), 1584 (C N), 1107 (C–O–C);
–
–
–
–
–
2
2
CDCl3): d 8.71 (brs, 1H, SH), 7.57–7.54 (t, J ¼ 8.4 Hz, 2H), 7.54–7.52
(d, J ¼ 6.9 Hz, 2H), 7.44–7.39 (m, 6H), 7.32–7.29 (t, J ¼ 8.1 Hz, 1H),
7.06–7.03 (d, J ¼ 8.7 Hz, 2H), 4.52 (s, 2H, O–CH2). Maldi-MS (m/z):
393 (Mþ), 394 (Mþ1); Anal. Calcd. for C21H16ClN3OS: C, 64.03; H,
4.09; N, 10.67. Found: C, 64.08; H, 4.12; N, 10.63%.
1H NMR (400 MHz, CDCl3) d 7.90–7.88 (d, J ¼ 6.2 Hz, 2H), 7.58–7.60
(d, J ¼ 7.8 Hz, 2H), 7.50–7.45 (m, 6H, Ar–H), 7.39–7.28 (m, 6H,
Ar–H), 6.94–6.92 (d, J ¼ 6.4 Hz, 2H), 5.08 (s, 2H, O–CH2), 4.51
(s, 2H, S–CH2); 13C NMR (100 MHz, DMSO-d6) d 191.85, 156.34,
148.47, 145.75, 136.46, 135.18, 132.76, 131.05, 130.27, 129.67,
129.18, 128.98, 128.99, 127.85, 128.67, 127.24, 126.88, 114.98,
60.07, 36.96; Maldi-MS (m/z): 512 (Mþ), 513 (Mþ1); Anal. Calcd.
for C29H22ClN3O2S: C, 68.03; H, 4.33; N, 8.21. Found: C, 68.09; H,
4.30; N, 8.19.
5-[(Biphenyl-4-yloxy)methyl]4-(4-nitrophenyl)-3-mercapto-(4H)-1,2,4-triazole
(5b) Yield 89%; light yellow crystals; m.p. 221–2228C; IR (KBr) cmꢀ1
:
2912 (CH ), 2690 (S–H), 1604 (C N), 1102 (C–O). H NMR (300 MHz,
1
–
–
2
CDCl3) d 8.93 (brs, 1H, S-H), 8.39 (d, J ¼ 8.6 Hz, 2H,), 7.65
(d, J ¼ 8.8 Hz, 2H), 7.52–7.32 (m, 6H), 7.28 (t, J ¼ 6.9 Hz, 1H),
6.96 (d, J ¼ 6.6 Hz, 2H), 4.81 (s, 2H, O–CH2). Maldi-MS (m/z): 404
(Mþ), 405 (Mþ1). Anal. Calcd. for C21H16N4O3S: C, 62.36; H, 3.99; N,
13.85. Found: C, 62.39; H, 4.01; N, 12.79%.
2-{5-[(4-Biphenyloxy)methyl]-4-(4-chlorophenyl)-4H-1,2,4-triazol-3-ylthio}-1-
(4-methoxyphenyl) ethanone (6b) Yield 75%; white crystals; m.p.
178–1808C; IR (KBr) cmꢀ1: 2912 (CH ), 1681 (C O), 1607 (C N),
–
–
–
–
2
1093 (C–O–C); 1H NMR (400 MHz, CDCl3) d 7.86–7.84 (d, J ¼ 6.4 Hz,
2H), 7.62–7.60 (d, J ¼ 6.4 Hz, 2H), 7.46–7.51 (m, 6H), 7.33–7.27 (m,
3H), 6.96–6.91 (dd, J ¼ 6.6 Hz, 4H), 5.01 (s, 2H, O–CH2), 4.53 (s, 2H,
S–CH2), 3.83 (s, 3H, O–CH3); 13C NMR (100 MHz, DMSO-d6) d 192.94,
156.78, 156.34, 148.47, 145.75, 135.16, 134.78, 131.05, 130.26,
130.16, 129.57, 128.96, 126.88, 129.86, 128.95, 127.85, 127.24,
117.56, 114.98, 60.07, 36.96; Maldi-MS (m/z): 542 (Mþ), 543
(Mþ1); Anal. Calcd. for C30H24ClN3O3S: C, 66.47; H, 4.46; N,
7.75. Found: C, 66.50; H, 4.47; N, 7.72.
5-[(Biphenyl-4-yloxy)methyl]4-(4-methoxyphenyl)-3-mercapto-(4H)-1,2,4-
triazole (5c) Yield 68%; white crystals; m.p. 168–1708C; IR (KBr)
1
cmꢀ1: 3058 (Ar–H), 2688 (S–H), 1603 (C N), 1107 (C–O); H NMR
–
–
(300 MHz, CDCl3): d 8.71 (brs, 1H, SH), 7.52 (d, J ¼ 5.4 Hz, 2H),
7.49 (d, J ¼ 6.9 Hz, 2H), 7.42 (d, J ¼ 5.7 Hz, 2H), 7.36–7.30
(m, 3H), 7.01 (d, J ¼ 6.6 Hz, 2H), 6.91 (d, J ¼ 6.6 Hz, 2H), 4.93
(s, 2H, O-CH2), 3.88 (s, 3H, O–CH3); Maldi-MS (m/z): 389 (Mþ), 390
(Mþ1); Anal. Calcd. for C22H19N3O2S: C, 67.84; H, 4.92; N, 10.79.
Found: C 67.76; H 4.89; N 10.75.
2-{5-[(4-Biphenyloxy)methyl]-4-(4-chlorophenyl)-4H-1,2,4-triazol-3-ylthio}-1-
(4-bromophenyl)ethanone (6c) Yield 68%; white crystals; m.p.
5-[(Biphenyl-4-yloxy)methyl]4-(phenyl)-3-mercapto-(4H)-1,2,4-triazole (5d)
Yield 78%; white crystals; m.p. 234–2368C; IR (KBr) cmꢀ1: 2912
154–1568C; IR (cmꢀ1): 2912 (CH ), 1683 (C O), 1607, 1584
–
–
2
1
(CH ), 2690 (S–H), 1607 (C N), 1110 (C–O–C); H NMR (300 MHz,
(C N), 1093 (C–O–C); 1H NMR (400 MHz, CDCl3)
d
7.88
–
–
–
–
2
CDCl3) d 9.57 (brs, 1H, S–H), 7.57–7.52 (m, 4H), 7.44–7.25 (m, 8H),
7.04 (2H, d, J ¼ 8.7 Hz), 4.69 (s, 2H, O–CH2); Maldi- MS (m/z): 359
(Mþ), 360 (Mþ1); Anal. Calcd. for C21H17N3OS: C, 70.17; H, 4.77; N,
11.69. Found: C, 70.09; H, 4.64; N, 11.58.
(d, J ¼ 6 Hz, 2H), 7.62 (d, J ¼ 6 Hz, 2H), 7.51–7.46 (m, 6H), 7.39
(t, J ¼ 5.7 Hz, 2H), 7.34 (d, J ¼ 6.3 Hz, 2H), 7.29 (t, J ¼ 5.7 Hz, 1H),
6.94 (d, J ¼ 6.3 Hz, 2H), 5.12 (s, 2H, O-CH2), 4.92 (s, 2H, S–CH2);
13C NMR (100 MHz, CDCl3) d 192.64, 156.95, 148.24, 145.25,
142.00, 135.17, 134.76, 130.85, 130.26, 130.06, 129.35, 129.27,
128.87, 128.82, 128.76, 127.65, 126.92, 126.88, 115.18, 59.96,
35.87; Maldi-MS (m/z): 590 (Mþ), 591 (Mþ1); Anal. Calcd.
for C29H21ClBrN3O2S: C, 58.94; H, 3.58; N, 7.11. Found: C, 59.02
H, 3.59; N, 7.08.
5-[(Biphenyl-4-yloxy)methyl]4-(4-fluorophenyl)-3-mercapto-(4H)-1,2,4-triazole
(5e) Yield 76%; white crystals; m.p. 186–1888C; IR (KBr) cmꢀ1: 3049
–
(Ar–H), 2929 (CH ), 2680 (S–H), 1603, 1514 (C N), 1105 (C–O);
–
2
1H NMR (300 MHz, CDCl3) d 8.73 (brs, 1H, SH), 7.51–7.46
(m, 4H), 7.41–7.37 (m, 4H), 7.28 (t, J ¼ 5.4 Hz, 1H), 7.21
(t, J ¼ 8.3 Hz, 2H), 6.94 (d, J ¼ 6.3 Hz, 2H), 4.73 (s, 2H, O–CH2);
Maldi-MS (m/z): 377 (Mþ), 378 (Mþ1); Anal. Calcd. for C21H16FN3OS:
C, 66.83; H, 4.27; N, 11.13. Found: C, 66.92; H, 4.25; N, 11.06.
5-[(Biphenyl-4-yloxy)methyl]4-(2-methylphenyl)-3-mercapto-(4H)-1,2,4-
triazole (5f) Yield 72%; white crystals; m.p. 186–1888C; IR
2-{5-[(4-Biphenyloxy)methyl]-4-(4-chlorophenyl)-4H-1,2,4-triazol-3-ylthio}-1-
(4-nitrophenyl)ethanone (6d) Yield 72%; light yellow crystals; m.p.
183–1848C; IR (KBr) cmꢀ1: 2905 (C–H), 1684 (C O), 1605, 1525
–
–
(C N), 1320 (N O), 1093 (C–O–C); H NMR (400 MHz, CDCl ) d 8.34
1
–
–
–
–
3
(d, J ¼ 6.6 Hz, 2H), 8.20 (d, J ¼ 6.6 Hz, 2H), 7.53–7.47 (m, 6H), 7.41
(t, J ¼ 5.7Hz; 2H), 7.37 (d, J ¼ 6.6 Hz, 2H), 7.31 (t, J ¼ 5.7 Hz, 1H),
6.95 (d, J ¼ 6.3 Hz, 2H), 5.10 (s, 2H, O–CH2), 4.91 (s, 2H, S–CH2);
(cmꢀ1): 2934 (CH ), 2686 (S–H), 1604, 1594 (C N), 1108
–
–
2
(C–O–C). 1H NMR (300 MHz, CDCl3): d 8.31 (brs, 1H, S–H),
ß 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
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210
M. M. Alam et al.
Arch. Pharm. Chem. Life Sci. 2012, 345, 203–214
13C NMR (100 MHz, DMSO-d6) d 191.65, 156.81, 150.00, 147.00,
140.41, 139.72, 136, 135.17, 131.12, 130.92, 130.28, 128.78, 128.31,
126.97, 126.79, 124.09, 115.10, 59.87, 36.75; Maldi-MS (m/z) 557
(Mþ), 558 (Mþ1); Anal. Calcd. for C29H21ClN4O4S: C, 62.53; H, 3.80;
N, 10.06. Found: C, 62.62; H, 3.82; N, 9.96.
147.37, 137.26, 135.87, 134.83, 133.20, 132.27, 130.76, 129.05,
128.92, 128.56, 128.09, 127.12, 126.79, 115.12, 115.12, 114.94,
60.24, 56.14, 36.13; Maldi-MS (m/z) 507 (Mþ), 508 (Mþ1); Anal.
Calcd. for C30H25N3O3S: C, 70.98; H, 4.96; N, 8.28. Found: C,
71.02; H, 4.95; N, 8.25.
2-{5-[(4-Biphenyloxy)methyl]-4-(4-nitrophenyl)-4H-1,2,4-triazol-3-ylthio}-1-
(phenyl)ethanone (6e) Yield 66%; light yellow crystals; m.p.
2-{5-[(4-Biphenyloxy)methyl]-4-(4-methoxyphenyl)-4H-1,2,4-triazol-3-ylthio}-
1-(4-methoxyphenyl)ethanone (6j) Yield 78%; white crystals; m.p.
168–1708C; IR (KBr) cmꢀ1: 2907 (CH ), 1686 (C O), 1515, 1524
157–1598C; IR (KBr) cmꢀ1: 2907 (CH ), 1687 (C O), 1586 (C N),
–
–
–
–
–
–
2
2
(C N), 1093 (C–O–C); 1H NMR (400 MHz, CDCl3)
d
8.41
1087 (C–O–C); 1H NMR (400 MHz, CDCl3) d 7.99 (d, J ¼ 9.0 Hz, 2H),
7.67 (d, J ¼ 8.9 Hz, 2H), 7.52–7.46 (m, 4H), 7.41 (t, J ¼ 7.5 Hz, 2H),
7.31 (t, J ¼ 5.8 Hz, 1H), 7.08–6.88 (m, 6 H), 5.15 (s, 2H, O–CH2),
4.92 (s, 2H, S–CH2), 3.87 (s, 6H, O–CH3); 13C NMR (100 MHz,
–
–
(d, J ¼ 8.7 Hz, 2H), 7.98 (d, J ¼ 9 Hz, 2H), 7.66 (d, J ¼ 8.9 Hz,
2H), 7.52–7.28 (m, 10H), 6.96 (d, J ¼ 6.6 Hz, 2H), 5.14 (s, 2H,
O–CH2), 4.91 (s, 2H, S–CH2); 13C NMR (100 MHz, CDCl3) d
191.24, 156.88, 150.12, 147.34, 141.46, 138.56, 135.86, 135.76,
130.68, 130.27, 128.65, 128.32, 128.31, 126.79, 124.09, 122.34,
115.17, 59.86, 35.79; MS (m/z): 522 (Mþ), 523 (Mþ1); Anal. Calcd.
for C29H22N4O4S: C, 66.65; H, 4.24; N, 10.72. Found: C, 66.72; H,
4.26; N, 10.68.
CDCl3)
d 191.94, 164.56, 156.82, 160.68, 150.19, 147.38,
135.14, 134.54, 130.96, 129.96, 129.07, 128.92, 128.56, 127.13,
127.10, 126.80, 115.13, 114.94, 114.92, 60.25, 55.63, 37.10;
Maldi-MS (m/z): 537 (Mþ), 538 (Mþ1); Anal. Calcd. for
C31H27N3O4S: C, 69.25; H, 5.06; N, 7.82. Found: C, 69.30; H, 5.02;
N, 7.79.
2-{5-[(4-Biphenyloxy)methyl]-4-(4-nitrophenyl)-4H-1,2,4-triazol-3-ylthio}-
1-(4-phenylphenyl)ethanone (6f) Yield 70%, light yellow crystals, m.p.
2-{5-[(4-Biphenyloxy)methyl]-4-(phenyl)-4H-1,2,4-triazol-3-ylthio}-1-(4-
bromophenyl) ethanone (6k) Yield 72%; white crystals; m.p.
175–1778C; IR (KBr) cmꢀ1: 2907 (C–H), 1671 (C O), 1601, 1558
–
–
(C N), 1093 (C–O–C); 1H NMR (300 MHz, CDCl3)
d
8.41
164–1668C; IR (cmꢀ1): 2907 (CH ), 1686 (C O), 1570 (C N),
–
–
–
–
–
–
7.83
2
(d, J ¼ 8.4 Hz, 2H), 8.10–8.07 (d, J ¼ 7.8 Hz, 2H), 7.73–7.63
(m, 6H), 7.52–7.39 (m, 8H), 7.31 (t, J ¼ 5.7 Hz, 1H), 6.93
(d, J ¼ 7.8 Hz, 2H), 5.15 (s, 2H, O–CH2), 5.00 (s, 2H, S–CH2);
13C NMR (100 MHz, CDCl3) d 191.34, 191.28, 156.98, 150.14,
147.33, 141.47, 138.86, 138.57, 135.84, 135.38, 135.12, 130.73,
130.70, 130.69, 130.29, 128.37, 128.32, 127.34, 126.87, 126.80,
124.09, 122.36, 115.16, 61.09, 37.06; Maldi-MS (m/z): 598 (Mþ) 599
(Mþ1); Anal. Calcd. for C35H26N4O4S: C, 70.22; H, 4.38; N, 9.36.
Found: C, 70.14; H, 4.35; N, 9.34.
1093 (C–O–C); 1H NMR (400 MHz, CDCl3)
d
(d, J ¼ 6.0 Hz, 2H), 7.56 (d, J ¼ 6.3 Hz, 2H), 7.41–7.34
(m, 11H), 7.23 (t, J ¼ 5.4 Hz, 1H), 6.87 (d, J ¼ 6.3 Hz, 2H),
5.02 (s, 2H, O–CH2), 4.97 (s, 2H, S–CH2); 13C NMR (100 MHz,
CDCl3) d 192.19, 156.99, 150.17, 147.37, 135.14, 134.91, 132.52,
132.24, 130.07, 129.98, 128.78, 128.26, 126.95, 126.91, 126.81,
126.79, 115.18, 63.35, 41.01; Maldi-MS (m/z): 556 (Mþ), 557
(Mþ1); Anal. Calcd. for C29H22BrN3O2S: C, 62.59; H, 3.98; N,
7.55. Found: C, 62.64; H, 4.01; N, 7.52.
2-{5-[(4-Biphenyloxy)methyl]-4-(4-methoxyphenyl)-4H-1,2,4-triazol-3-ylthio}-
1-(4-bromophenyl)ethanone (6g) Yield 70%; white crystals; m.p.
2-{5-[(4-Biphenyloxy)methyl]-4-(phenyl)-4H-1,2,4-triazol-3-ylthio}-1-(4-nitro-
phenyl)ethanone (6l) Yield 67%; light yellow crystals; m.p.
156–1588C; IR (KBr) cmꢀ1: 2907 (C–H), 1686 (C O), 1605, 1536
170–1728C; IR (cmꢀ1): 2907 (CH ), 1684 (C O), 1541 (C N),
–
–
–
–
–
–
2
(C N), 1105 (C–O–C); 1H NMR (CDCl3) d 7.53–7.48 (m, 4H), 7.51–
1107 (C–O–C); 1H NMR (400 MHz, CDCl3) d 8.34 (d, J ¼ 6.6Hz,
2H), 8.22 (d, J ¼ 6.6 Hz, 2H), 7.56–7.54 (m, 3H), 7.51 (d, J ¼ 6.3Hz,
2H), 7.47 (d, J ¼ 6.6 Hz, 2H), 7.42–7.39 (m, 4H), 7.30 (t, J ¼ 5.4 Hz,
1H), 6.95 (d, J ¼ 6.3 Hz, 2H), 5.10 (s, 2H, O–CH2), 4.92 (s, 2H,
S–CH2); 13C NMR (100 MHz, CDCl3) d 191.88, 156.95, 152.22,
150.73, 140.46, 139.75, 134.97, 132.43, 130.53, 130.03, 129.72,
128.78, 128.27, 126.92, 126.78, 124.09, 115.17, 59.92, 39.86;
TOF-MS (m/z): 522 (Mþ), 523 (Mþ1); Anal. Calcd. for
C29H22N4O4S: C, 66.65; H, 4.24; N, 10.72. Found: C, 66.69; H,
4.23; N, 10.6.
–
–
7.37 (m, 8H), 7.28 (t, J ¼ 5.5 Hz, 1H), 7.24–7.18 (m, 2H), 7.01
(d, J ¼ 6.7 Hz, 2H), 6.90 (d, J ¼ 6.6 Hz, 2H), 5.15 (s, 2H, O–CH2),
4.85 (s, 2H, S–CH2), 3.88 (s, 3H, O–CH3); 13C NMR (CDCl3) d 191.45,
160.65, 156.84, 150.25, 147.34, 140.34, 135.89, 134.81, 132.24,
130.65, 129.08, 128.96, 128.84, 128.74, 128.56, 127.22, 126.98,
126.80, 122.48, 114.94, 60.09, 37.58; Maldi-MS (m/z): 586 (Mþ),
587 (Mþ1); Anal. Calcd. for C30H24BrN3O3S: C, 61.44; H, 4.12; N,
7.16. Found: C, 61.48; H, 4.09; N, 7.13.
2-{5-[(4-Biphenyloxy)methyl)-4-(4-methoxyphenyl)-4H-1,2,4-triazol-3-ylthio}-
1-(4-chlorophenyl)ethanone (6h) Yield 79%; white crystals; m.p.
3-(4-Nitrophenylethylthio)-5-[(4-biphenyloxy)methyl]-4-phenyl-4H-1,2,4-
triazole (6m) Yield 74%; light yellow crystals; m.p. 138–1408C; IR
154–1568C; IR (KBr) cmꢀ1: 2907 (C–H ), 1655 (C O), 1605 (C N),
–
–
–
–
2
1239 (C C), 1093 (C–O–C); 1H NMR (400 MHz, CDCl3) d 7.90
(KBr) cmꢀ1: 2907 (CH ), 1525 (C N), 1128 (C–O–C); 1H NMR
–
–
–
–
2
(d, J ¼ 8.4 Hz, 2H), 7.52–7.46 (m, 4H), 7.40 (t, J ¼ 7.5 Hz, 2H),
7.32–7.25 (m, 5H), 7.00 (d, J ¼ 9 Hz, 2H), 6.96 (d, J ¼ 8.4 Hz, 2H),
5.12 (s, 2H, O–CH2), 4.91 (s, 2H,S–CH2), 3.85 (s, 3H,O–CH3); 13C NMR
(100 MHz, CDCl3) d 190.24, 160.69, 156.84, 150.15, 147.35, 140.21,
137.87, 135.87, 134.81, 132.24, 130.73, 129.03, 128.99, 128.57,
128.08, 127.12, 126.81, 115.27, 114.94, 60.12, 38.03, 55.35;
Maldi-MS (m/z): 542 (Mþ), 543 (Mþ1); Anal. Calcd. for
C30H24ClN3O3S: C, 66.47; H, 4.46; N, 7.75. Found: C, 66.53; H,
4.44; N, 7.69.
(400 MHz, CDCl3)
d
8.12 (d, J ¼ 6.0 Hz, 2H), 7.56–7.47
(m, 8H), 7.41 (d, J ¼ 7.2 Hz, 2H), 7.30 (t, J ¼ 5.4 Hz, 1H), 7.23
(d, J ¼ 6.6 Hz, 2H), 6.94 (d, J ¼ 5.7 Hz, 2H), 5.09 (s, 2H, O–CH2),
4.53 (s, 2H, S–CH2), 3.74 (t, J ¼ 8.4 Hz, 2H), 3.12 (t, J ¼ 8.0 Hz,
2H); 13C NMR (100 MHz, CDCl3) d 156.90, 148.45, 147.43, 144.24,
140.45, 135.92, 130.43, 130.11, 130.45, 129.45, 129.45, 128.77,
126.88, 123.82, 115.17, 59.87, 35.87; Maldi-MS (m/z): 508 (Mþ),
509 (Mþ1); Anal. Calcd. for C29H24N4O3S: C, 68.49; H, 4.76; N,
11.02. Found: C, 68.55; H, 4.74; N, 11.05.
2-{5-[(4-Biphenyloxy)methyl]-4-(4-methoxyphenyl)-4H-1,2,4-triazol-3-ylthio}-
1-(phenyl)ethanone (6i) Yield 82%, white crystals, m.p. 150–1528C; IR
3-(Phenylethylthio)-5-[(4-biphenyloxy)methyl]-4-phenyl-4H-1,2,4-triazole (6n)
Yield 70%; m.p. 138–1408C; IR (KBr) cmꢀ1: 2907 (CH2), 1541, 1523
1
(C N), 1107 (C–O–C); H NMR (400 MHz, CDCl ) d 7.52–7.49 (6H, m),
(KBr) cmꢀ1: 2907 (CH ), 1686 (C O), 1541 (C N), 1102 (C–O–C);
–
–
–
–
–
–
2
3
1H NMR (400 MHz, CDCl3) d 7.91 (d, J ¼ 8.4 Hz, 2H,), 7.46–7.40
(m, 6H), 7.40 (t, J ¼ 7.5 Hz, 2H), 7.32–7.25 (m, 6H), 6.96
(d, J ¼ 8.4 Hz, 2H), 5.10 (s, 2H, O–CH2), 4.92 (s, 2H, S–CH2), 3.85
(s, 3H, O–CH3); 13C NMR (CDCl3) d 190.20, 160.69, 156.81, 150.17,
7.40 (2H, t, J ¼ 7.2 Hz), 7.34–7.31 (m, 3H), 7.25–7.18 (m, 6H), 6.96
(2H, d, J ¼ 9Hz), 5.10 (2H, s, OCH2), 3.50 (2H, t, J ¼ 7.8 Hz, S–CH2),
3.08 (2H, t, J ¼ 7.2 Hz); 13C NMR (100 MHz, CDCl3) d 156.88, 148.79,
148.34, 145.56, 142.43, 134.91, 130.45, 130.43, 130.34, 130.04,
ß 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
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Arch. Pharm. Chem. Life Sci. 2012, 345, 203–214
1,2,4-Triazole Derivatives as Potent Bio-Active Agents
211
129.98, 128.79, 128.76, 128.77, 126.88, 124.24, 123.89, 115.16,
59.84, 36.74, 35.86; Maldi-MS (m/z): 522 (Mþ), 523 (Mþ1); Anal.
Calcd. for C29H22N4O4S: C, 66.65; H, 4.24; N, 10.72. Found: C,
66.60; H, 4.23; N, 10.69.
General procedure for synthesis of 4-(substituted phenyl)-
3-(amine-4-ylmethylthio)-5-(4-biphenyloxymethyl)-4H-
1,2,4-triazole (7a–7e)
To 50 mL absolute alcohol was added appropriate triazole (5b, 5f
and 5g) (5 mM), formaldehyde (5 mM) and appropriate amine
(5 mM), the reaction mixture was stirred for 24 h, and allowed to
stand overnight in refrigerator. The precipitate so obtained was
filtered, washed with cold alcohol and recrystallized from
alcohol.
2-{5-[(4-Biphenyloxy)methyl]-4-(4-fluorophenyl)-4H-1,2,4-triazol-3-ylthio}-
1-(4-bromophenyl)ethanone (6o) Yield 72%; white crystals; m.p. 160–
1628C; IR (cmꢀ1): 2907 (CH ), 1684 (C O), 1605 (C N), 1108
–
–
–
–
2
(C–O–C); 1H NMR (400 MHz, CDCl3) d 6.92–7.90 (m, 17 H,
Ar–H), 5.08 (s, 2H, O–CH2), 4.92 (s, 2H, S–CH2); 13C NMR
(100 MHz, CDCl3) d 192.34, 162.98, 156.87, 145.69, 145.58,
137.24, 134.89, 131.29, 130.87, 130.76, 130.76, 130.46, 130.41,
130.09, 128.78, 126.87, 126.79, 126.23, 115.15, 114.97, 59.85,
36.79; Maldi-MS (m/z): 574 (Mþ), 575 (Mþ1); Anal. Calcd.
for C29H21BrFN3O2S: C, 60.63; H, 3.68; N, 7.31. Found: C, 60.67;
H, 3.69; N, 7.32.
5-[(4-Biphenyloxy)methyl]-4-(4-chlorophenyl)-2-(morpholinomethyl)-2H-1,2,4-
triazole-3(4H)-thione (7a) Yield 58%; white crystals; m.p. 180–1828C; IR
(KBr) cmꢀ1: 2909 (C–H), 1608 (C N), 1381 (C S), 1115 (C–O–C);
–
–
–
–
7.86–7.29 (m, 11H), 6.89
1H NMR (300 MHz, CDCl3)
d
(d, J ¼ 8.7 Hz, 2H), 5.18 (s, 2H, O–CH2), 4.93 (s, 2H, N–CH2–N),
3.72 (t, J ¼ 3.9 Hz, 4H), 2.86 (t, J ¼ 4.5 Hz 4H, N-(CH2)2);
13C NMR (100 MHz, CDCl3) d 170.72, 156.41, 146.12, 140.22,
136.29, 135.51, 132.08, 129.95, 129.26, 128.34, 129.64, 127.03,
126.76, 115.11, 69.86, 66.81, 59.99, 50.74; Maldi-MS (m/z): 493
(Mþ), 495 (Mþ2); Anal. Calcd. for C26H25N4O2S: C, 63.34; H, 5.11;
N, 11.36. Found: C, 63.52; H, 5.18; N, 11.41.
2-{5-[(4-Phenylphenoxy)methyl]-4-(4-fluorophenyl)-4H-1,2,4-triazol-3-ylthio}-
1-(phenyl)ethanone (6p) Yield 74%; white crystals; m.p. 156–1588C; IR
1
(cmꢀ1): 2933 (CH ), 1698 (C O), 1515 (C N), 1105 (C–O–C); H NMR
–
–
–
–
2
(300 MHz, CDCl3) d 7.89 (d, J ¼ 6.3 Hz, 2H), 7.63 (d, J ¼ 6 Hz, 2H),
7.51–7.46 (m, 5H), 7.41–7.37 (m, 4H), 7.29 (t, J ¼ 5.4 Hz, 1H), 7.41
(t, J ¼ 8.4Hz, 2H), 6.93 (d, J ¼ 6.3Hz, 2H), 5.08 (s, 2H, O–CH2), 4.92
(s, 2H, S–CH2); 13C NMR (100 MHz, CDCl3) d 191.36, 162.97, 156.78,
145.63, 145.27, 137.15, 136.15, 134.34, 133.89, 130.88, 130.87,
130.67, 130.45, 130.12, 130.10, 129.89, 128.77, 126.25, 115.13,
114.79, 58.97, 37.03; Maldi-MS (m/z): 495 (Mþ), 496 (Mþ1); Anal.
Calcd. for C29H22ClFN3O2S: C, 70.29; H, 4.47; N, 8.48. Found: C,
70.28; H, 4.43; N, 8.50.
5-[(4-Biphenyloxy)methyl]-(4-methoxyphenyl)-3-(N-methyl-piperazine-4-
ylmethylthio)-4H-1,2,4-triazole (7b) Yield 54%; white crystals; m.p.
184–1868C; IR (cmꢀ1): 2907 (C–H), 1557 (C N), 1326 (C S), 1109
–
–
–
–
(C–O–C); 1H NMR (200 MHz, CDCl3) d 7.60–7.36 (m, 9H), 7.11–
6.96 (m, 4H), 5.07 (s, 2H O–CH2), 4.49 (s, 2H, N–CH2–N), 3.81
(s, 3H, O–CH3), 2.71 (t, J ¼ 7.8 Hz 4H), 2.51 (t, J ¼ 9 Hz, 4H), 2.25
(s, 3H, N–CH3); 13C NMR (100 MHz, CDCl3) d 171.82, 163.49,
155.64, 156.68, 149.66, 135.69, 130.29, 129.64, 129.61, 128.69,
127.89, 127.62, 126.92, 115.45, 68.82, 68.45, 56.24, 49.26, 44.14,
57.15; Maldi-MS (m/z) 501 (Mþ), 502 (Mþ1); Anal. Calcd.
for C28H31N5O2S: C, 67.04; H, 6.23; N, 13.96. Found: C, 67.01;
H, 6.21; N, 13.93.
2-{5-[(4-Biphenyloxy)methyl]-4-(4-fluorophenyl)-4H-1,2,4-triazol-3-ylthio}-
1-(4-methoxyphenyl) ethanone (6q) Yield 85%, white crystals, m.p.
172–1748C; IR (cmꢀ1): 2907 (CH ), 1686 (C O), 1605 (C N),
–
–
–
–
2
1093 (C–O–C); 1H NMR (400 MHz, CDCl3) d 6.87–7.89 (m, 18 H),
5.14 (s, O–CH2, 2H), 4.91 (s, S–CH2, 2H), 3.87 (s, 3H, O–CH3);
13C NMR (100 MHz, CDCl3) d 192.05, 162.97, 161.67, 156.87,
146.00, 145.27, 133.91, 130.89, 130.26, 130.15, 130.12, 130.11,
129.87, 128.00, 126.26, 115.14, 114.96, 114.78, 114.15, 58.98,
37.47, 55.87; Maldi-MS (m/z): 525 (Mþ), 526 (Mþ1); Anal. Calcd.
for C30H24FN3O3S: C, 68.56; H, 4.60; N, 7.99. Found: C, 68.60; H,
4.58; N, 8.01.
5-[(4-Biphenyloxy)methyl]-4-(4-methoxyphenyl)-2-[(pyrrolidin-1-yl)methyl]-
2H-1,2,4-triazole-3(4H)-thione (7c) Yield 80%; white crystals; m.p. 174–
1768C; IR (KBr) cmꢀ1: 2907 (C–H), 1607, 1516 (C N), 1316 (C S),
–
–
–
–
1104 (C–O–C); 1H NMR (DMSO-d6) d 6.91–7.54 (m, 14H, Ar–H), 5.22
(s, 2H, O–CH2), 4.47 (s, 2H, N–CH2–N), 3.82 (s, 3H, O–CH3), 2.24
(t, J ¼ 8 Hz, 4H, N–CH2), 1.53 (t, J ¼ 7.5 Hz, 4H, CH2–CH2);
13C NMR (100 MHz, DMSO-d6) d 172.86, 163.54, 155.76, 149.68,
135.65, 133.89, 130.27, 129.67, 129.58, 128.89, 128.69, 127.79,
126.89, 126.75, 115.42, 68.76, 68.45, 52.42, 26.41; Maldi-MS (m/z):
472 (Mþ), 473 (Mþ1); Anal. Calcd. for C27H28N4O2S: C, 68.62; H,
5.97; N, 11.85. Found: C, 68.68; H, 5.99; N, 11.87.
3-(Phenylethylthio)-5-[(4-biphenyloxy)methyl]-4-(4-fluorophenyl)-4H-1,2,4-
triazole (6r) Yield 62%; white crystals; m.p. 144–1468C; IR (KBr)
cmꢀ1: 2907 (C–H), 1520, 1517 (C N), 1108 (C–O–C); 1H NMR
–
–
(300 MHz, CDCl3): d 7.52–7.49 (m, 6H), 7.40 (t, J ¼ 7.8 Hz, 2H),
7.34–7.18 (m, 9H), 6.96 (d, J ¼ 9.0 Hz, 2H), 5.08 (s, 2H, O–CH2),
3.50 (t, J ¼ 7.5 Hz, 2H, S–CH2), 3.08 (t, J ¼ 8.1 Hz, 2H,); 13C NMR
(100 MHz, CDCl3) d 161.69, 156.98, 149.00, 147.18, 145.14,
135.14, 134.79, 130.98, 130.45, 130.25, 130.08, 130.03, 129.85,
129.85, 129.17, 127.99, 123.07, 114.78, 114.56, 59.12, 37.49,
55.58; Maldi-MS (m/z): 481 (Mþ), 482 (Mþ1); Anal. Calcd.
for C29H24FN3OS: C, 72.33; H, 5.02; N, 8.73. Found: C, 72.29; H,
5.04; N, 8.69.
5-[(4-Biphenyloxy)methyl]-4-(phenyl)-2-(morpholinomethyl)-2H-1,2,4-
triazole-3(4H)-thione (7d) Yield 64%; white crystals; m.p. 132–1348C;
IR (cmꢀ1): 2912 (CH ), 1607 (C N), 1324 (C S), 1103 (C–O–C);
–
–
–
–
2
1H NMR (200 MHz, DMSO-d6) d 7.54–7.40 (m, 12H), 6.93 (d,
J ¼ 8.8 Hz, 2H), 5.22 (s, 2H, O–CH2), 4.97 (s, 2H, N–CH2–N),
4.76 (t, J ¼ 6.7 Hz, 4H), 2.90 (t, J ¼ 7.0 Hz, 4H); 13C NMR
(100 MHz, DMSO-d6) d 170.56, 163.49, 155.76, 149.68, 135.65,
133.89, 130.27, 129.67, 129.58, 128.89, 128.69, 127.79, 126.89,
115.42, 115.42, 64.86, 68.76, 68.45, 64.86, 49.23; Maldi-MS (m/z):
458 (Mþ), 459 (Mþ1); Anal. Calcd. for C26H26N4O2S: C, 68.10; H,
5.71; N, 12.22. Found: C, 68.07; H, 5.69; N, 12.20.
3-(4-Nitrophenylmethylthio)-5[{(4-biphenyloxy)methyl]-4-(4-fluorophenyl)-
4H-1,2,4-triazole (6s) Yield 68%; light yellow crystals; m.p. 168–
1728C; IR (KBr) cmꢀ1: 2916 (C–H), 1524 (C N), 1106 (C–O–C);
–
–
1H NMR (400 MHz, CDCl3) d 8.04 (2H, d, J ¼ 8.4 Hz), 7.49–7.40
(m, 6H), 7.33 (t, J ¼ 7.6 Hz, 2H), 7.23 (t, J ¼ 5.7 Hz, 1H), 7.16–7.08
(m, 4H), 6.87 (d, J ¼ 8.4 Hz, 2H), 5.01 (s, 2H, O–CH2), 4.45 (s, 2H,
S–CH2); 13C NMR (100 MHz, CDCl3) d 162.97, 156.89, 155.76,
146.57, 146.23, 137.73, 134.91, 131.27, 130.89, 130.48, 130.27,
130.12, 129.89, 128.79, 126.89, 126.27, 123.27, 115.14, 115.14,
114.97, 59.89, 36.86; Maldi-MS (m/z): 512 (Mþ), 513 (Mþ1); Anal.
Calcd. for C28H21FN4O3S: C, 65.61; H, 4.13; N, 10.93. Found:
C, 65.67; H, 4.09; N, 10.90.
5-[(4-Biphenyloxy)methyl]-4-(2-methylphenyl)-2-(morpholinomethyl)-2H-
1,2,4-triazole-3(4H)-thione (7e) Yield 52%; white crystals; m.p. 126–
1288C; IR (cmꢀ1): 2917 (C–H), 1516 (C N), 1324 (C S), 1108
–
–
–
–
(C–O–C); 1H NMR (300 MHz, CDCl3) d 7.58–7.30 (m, 11H), 6.90
(d, J ¼ 8.7 Hz, 2H), 5.23 (s, 2H, O–CH2), 4.93 (s, 2H, N–CH2–N),
3.57 (t, J ¼ 6.7 Hz, 4H), 2.72 (t, J ¼ 6.9 Hz, 4H), 2.50 (s, 3H,
Ar–CH3); 13C NMR (100 MHz, DMSO-d6) d 170.58, 163.51,
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M. M. Alam et al.
Arch. Pharm. Chem. Life Sci. 2012, 345, 203–214
155.86, 149.66, 135.66, 130.26, 136.84, 134.43, 129.63, 129.57,
128.86, 128.81, 126.88, 126.98, 126.97, 115.43, 68.79, 68.56,
64.87, 49.24; Maldi-MS (m/z) 472 (Mþ), 473 (Mþ1); Anal. Calcd.
for C27H28N4O2S: C, 68.62; H, 5.97; N, 11.85. Found: C, 68.65; H,
5.95; N, 11.83.
Histopathological studies
For the histopathological study, rats were sacrificed under light
anesthesia after 4 h of the doses (3 times to the dose used for anti-
inflammatory) and their stomach specimens were removed and
put into 10% formalin solution. A longitudinal section of
stomach along the greater curvature, which included the ulcer
base and both sides of the ulcer margin, was taken and fixed in
10% formalin for 24 h at 48C and embedded in white solid
paraffin. Morphological examination was performed with
haematoxylin and eosin staining to analyze histological changes
and examined under electron microscope. The results are given
in Table 3 and Figure 1.
Biological screening
Albino Wistar rats of either sex (150–200 g) were obtained
from Central Animal House, Hamdard University, New Delhi.
The animals were kept in cages at room temperature and fed
with food and water ad libitum. Fourteen hours before the start of
the experiment the animals were sent to lab and fed only with
water ad libitum. The experiments were performed in accordance
with the rules of Institutional Animals Ethics Committee (regis-
tration number 173-CPCSEA). The animals were divided into six
groups with six animals in each group. Bacterial and fungal
cultures were obtained from Microbiology lab Batra Hospital,
New Delhi, India and the screening tests were carried out at the
Microbiology lab, Majidia hospital, Jamia Hamdard, New Delhi,
India.
In vitro antimicrobial activity studies
All bacterial and yeast strains were obtained from the Batra
Hospital, New Delhi, India. and Department of Biochemistry,
Jamia Hamdnd, New Delhi, India as follows: Pseudomonas
aeruginosa ATCC 27853, Klebsiella pneumoniae ATCC 700603,
Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923
and Candida albicans ATCC 10231, Candida krusei ATCC 6258,
Aspergillus niger MTCC 8189, Aspergillus flavus MTCC 277. All
the newly synthesized compounds were dissolved in dimethyl
sulfoxide (DMSO) to prepare chemicals of stock solution of
2 mg/1 mL.
Anti-inflammatory activity
All the synthesized compounds were evaluated for their anti-
inflammatory activity against carrageenan-induced acute paw
oedema in albino rats (Wistar strain) weighing 150–200 g [28–
29]. The animals were weighed and divided into control, stand-
ard, and test groups and each group contained six rats. The first
group of rats was treated with 0.1 mL of 0.5% CMC suspension
orally (control), second group was administered with a dose of
10 mg/kg of the suspension of ibuprofen (standard) and the test
group was treated with equimolar dose of the suspension of test
compounds relative to standard drug. After 30 min the animals
were injected with 0.1 mL of 1% carrageenan in normal saline
subcutaneously to the sub-plantar region of right hind paw. The
paw volume was measured at 0 h, 1 h, 3 h and 5 h, by using
plethysmometer. The amount of oedema in the drug-treated
groups was compared in relation to the control group with
the corresponding time intervals. The percent oedema inhibition
was calculated from the mean effect in the control and treated
animals according to the following equation: Percentage inhib-
ition ¼ 100 (1 – Vt/Vc), where Vt ¼ mean increase in paw volume
of test and Vc ¼ mean increase in paw volume of control group
of rats.
Agar-well diffusion method
Simple susceptibility screening test using agar-well diffusion
method [31] was used. Each microorganism was suspended in
Mueller Hinton (MH) (Difco, Detroit, MI) broth and diluted
approximately to 106 colony forming unit (cfu)/mL. They were
‘‘flood-inoculated’’ onto the surface of MH agar and Sabouraud
Dextrose Agar (SDA) and then dried. For Candida albicans, Candida
krusei, Aspergillus niger and Aspergillus flavus, SDA were used,
and for Pseudomonas aeruginosa, Klebsiella pneumoniae, Macconcy
agar was used and for Escherichia coli and Staphylococcus aureus
Muller Hinton agar was used. Six-millimeter diameter wells
were cut from the agar using a sterile cork-borer, and 200 mg
and 100 mg of the test compounds were delivered into the wells.
The plates were incubated for 18 h at 358C for bacteria and 48 h.
for fungal strains. Antimicrobial activity was evaluated by
measuring the zone of inhibition against the test organism.
Ampicillin (200, 100 mg) and fluconazole (200, 100 mg) were used
as standard drugs. Dimethyl sulfoxide was used as solvent
controls.
Analgesic activity
The analgesic activity was determined using tail flick method in
albino rats (Wistar strain) weighing 150–200 g [30]. The animals
were weighed and divided into control, standard, and test groups
and each group contained six rats. The first group of rats was
treated with 0.1 mL of 0.5% CMC suspension orally (control),
second group was administered with a dose of 10 mg/kg of
the suspension of ibuprofen (standard) and the test group was
treated with equimolar dose of the suspension of test compounds
relative to standard drug. After administration of drug reaction
time was measured at 0 min, 30 min, 60 min and 120 min by
immersing the tail in hot water (55 ꢁ 0.58C). Analgesic activity
was expressed as reaction time of tail flicking, compounds hav-
ing analgesic activity will show more reaction time than the
control (Table 2).
Statistical analysis
Statistical analysis was performed, followed by t test, to compare
different groups. P values less than 0.05 were considered to be
statistically significant.
The authors are thankful to Mr. A. Mueed, President, Hamdard National
Foundation (HNF) for providing financial support to Mahboob Alam
and Syed Nazreen. Authors also thank Vice-Chancellor, Jamia Hamdard
Dr. G. N. Qazi for providing necessary facilities, Regional Sophisticated
Instrumentation Centre, CDRI for spectral analysis of the compounds and
Dr. A. Mukherji, AIIMS, New Delhi for histopathological studies.
The authors have declared no conflict of interest.
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Arch. Pharm. Chem. Life Sci. 2012, 345, 203–214
1,2,4-Triazole Derivatives as Potent Bio-Active Agents
213
Figure 1. Haemtoxylin and eosin immunohistochemical staining of gastric ulcers after ulcer induction in rats.
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M. A. Alsheikh, H. S. Neau, Int. J. Med. Sci. 2010, 7, 232–
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