Synthesis, biological screening, in silico study and fingerprint applications of novel 1, 2, 4-triazole derivatives

Article abstract of DOI:10.1002/jhet.3929

A series of novel 1,2,4 triazole derivatives were synthesized by treating 4-bromo-2-(4H-1,2,4-triazole-3-yl)aniline (4) with different substituted benzene sulfonyl chlorides 5(a-f) and benzyl bromides 7(a-e). IR, ¹H-NMR, ¹³C-NMR, and

Full text of DOI:10.1002/jhet.3929

Received: 26 October 2019
DOI: 10.1002/jhet.3929
Revised: 23 January 2020
Accepted: 24 January 2020
A R T I C L E
Synthesis, biological screening, in silico study and
fingerprint applications of novel 1, 2, 4-triazole derivatives
Ghouse Khan1 | Swamy Sreenivasa1
| Shivaraja Govindaiah1
|
Vivek Chandramohan2 | Raghurama Shetty P1
1Department of Studies and Research in
Abstract
Organic Chemistry, Tumkur University,
Tumakuru, Karnataka, India
A series of novel 1,2,4 triazole derivatives were synthesized by treating
2Department of Biotechnology,
Siddaganga Institute of Technology,
Tumakuru, Karnataka, India
4-bromo-2-(4H-1,2,4-triazole-3-yl)aniline (4) with different substituted ben-
1
zene sulfonyl chlorides 5(a-f) and benzyl bromides 7(a-e). IR, H-NMR, 13C-
NMR, and mass analysis confirmed the structures of the newly synthesized
compounds. All derivatives were screened for their in vitro antibacterial activ-
ity against two bacterial strains viz Escherichia coli and Staphylococcus aureus,
antifungal activity against Aspergillus flavus and Candida albicans, anthelmin-
tic activity against Pheretima posthuma and also cytotoxicity activity against
MDA-MB 231 and A375 cancer cell lines. It was found that some of the deriva-
tives showed significant antibacterial, antifungal, anthelmintic, and cytotoxic
activities when compared to respective standard drugs. Molecular docking
studies have assisted the theoretical binding mode of the target molecules.
Compounds were also explored for fingerprint application.
Correspondence
Swamy Sreenivasa, Department of Studies
and Research in Organic Chemistry,
Tumkur University, Tumakuru,
Karnataka, India.
Email: dr.tumkuruniversity@gmail.com
1 | INTRODUCTION
attracting special interest because of their versatile phar-
macological properties due to their binding capacity to
People's migration from one location to another, climate
flexibility, air contamination, pollution of water, and so
forth can contribute to the spread of multiple microbial
diseases that pose a severe threat to human health. In the
late 1960s, there was a dramatic increase in the incidence
of fungal infections, which posed a global health risk.
The widespread use of azoles has resulted in serious resis-
tance, which has lowered their effectiveness considerably.
Recently, many drug-resistant human pathogenic
microbes have been reported.[1] To tackle the increasing
resistance of microorganisms, the scientific community
continues to be challenged to discover powerful and
selective antimicrobial agents. Similarly, anthelmintic
infections are becoming great cause of chronic illness in
tropical regions of the world. This disease became more
serious due to the resistance and accessibility of anthel-
mintics such as mebendazole, albendazole, and so forth.
In recent years, 1,2,4 triazole compounds have been
various target proteins.[2,3] This distinct triazole ring
structure made it easy to bind to various receptors and
enzymes in the biological system and exhibits vast biolog-
ical properties like antimicrobial, anti-inflammatory,
anticancer, anticonvulsant, antitumor, and anxiolytic.[4–7]
Nowadays, various triazole-based medicines have
been widely used in the clinic, such as antifungal
(Fluconazole, Voriconazole, and Itraconazole),[8] antican-
cer (Letrozole and Anastrozole)[9] as well as antiviral
(Ribavirin).[10] A large number of research articles have
been focused on synthesis and biological significance of
sulfonated and alkylated 1, 2, 4-triazole derivatives as
antibacterial,[11] anticancer,[11,12] carbonic anhydrase
inhibitor,[13]
analgesics,[14]
antihypertensive,[15–17]
in vitro antitrypanosomally and antileishmanial,[18] can-
nabinoid
receptor
binding,[19]
anticonvulsant
activity,[20,21] anticancer activity,[22,23] and so on. The ver-
satile utility of these derivatives prompted us to design
J Heterocyclic Chem. 2020;1–14.
wileyonlinelibrary.com/journal/jhet
© 2020 Wiley Periodicals, Inc.
1

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KHAN ET AL.
and synthesize novel derivatives of 1,2,4-triazole compris-
ing bromobenzene moiety and screened them for differ-
ent biological activities like antibacterial, antifungal,
anthelmintic and cytotoxic. The binding mode of the
active compounds was supported by molecular docking
study. We also report fingerprint applications of our
compounds.
which shows characteristic signals at 6.68 and 14.2 ppm
corresponding to NH2 and 1,2,4 triazole NH protons
respectively. Structure of this intermediate was also
established by 13C-NMR and Mass spectrum. Finally, the
intermediate (4) was treated with different substituted
benzene sulfonyl chlorides 5(a-f) and benzyl bromides 7
(a-e) to get two series of new compounds, sulfonamides 6
(a-f) and N-alkylated derivatives 8(a-e) containing 1,2,4
triazole moiety in good yield. These compounds were pre-
1
2 | RESULTS AND DISCUSSION
2.1 | Chemistry
pared as per the Scheme 1. The H NMR spectrum of 6
(a-f) series shows a characteristic singlet broad peak in
the range δ 5.1 to 6.8 ppm corresponds to NH proton of
sulfonamide linkage. NH proton signal of N-alkylated
derivatives 8(a-e) resonates in the range 4.41 to 6.6 ppm
whereas NH proton of 1,2,4 triazole nucleus of both the
series appeared as singlet in the range δ 8.7 to 9.45 ppm.
The aromatic protons of benzene and 1,2,4 triazole reso-
nated in the range δ 6.7 to 8.15 ppm. In 13C-NMR, ali-
phatic as well as aromatic carbons of both the series
appeared in the range 21 to 56 ppm and 106 to 160 ppm,
2-Amino-5-bromobenzoic acid on esterification afforded
ethyl 2-amino-5-bromobenzoate, which was converted to
ethyl 2-amino-5-bromobenzohydrazide (3) by treating
with hydrazine hydrate. Then the resulting compound
(3) was made to react with formamide to yield 4-bromo-
2-(1H-1,2,4-triazol-5-yl)aniline (4). The formation of this
compound was supported by the 1H-NMR spectrum
SCHEME 1 Reagents and conditions:
C2H5OH, H2SO4, 60ꢀC, 6 hours; NH2-NH2.2H2O,
EtOH, 60ꢀC, 8 hours; HCONH2, KOH, DMF,
200ꢀC, 3 hours; DCM, TEA, rt, 30 minutes;
MeCN, K2CO3, rt, 10 hours

KHAN ET AL.
3
respectively. More evidence has been established by
recording their mass spectra, which are in good agree-
ment with the calculated m/z values of respective
compounds.
6f ( NO2) with substituents at para position of phenyl
ring attached to sulfonamide linkage and 8a
(3,4-dichloro), 8b (2-Cl, 6-F), 8c ( CH3), 8d ( F), 8e
( CH3) with substituents of the benzyl ring exhibited
good activity. This activity may be due to electron-
withdrawing effect of NO2 group in case 6f and halogens
Cl, Br, F in compounds 6e, 8a, 8b, 8d, and 8e.[24] The
activity is pronounced in the case of 6f ( NO2) and 8b
(2-Cl, 6-F). The graphical representation of antibacterial
activity is given in Figure 1.
3 | BIOLOGICAL SCREENING
3.1 | Antibacterial activity
Synthesized compounds 6a-f and 8a-e were evaluated for
their in vitro antibacterial activity using disc diffusion
method against Staphylococcus aureus and Escherichia
coli bacterial strains. Ciprofloxacin was used as positive
control. The results of the antibacterial activity of com-
pounds 6a-f and 8a-e were reported in Table 1. The
results indicated that compounds show varying degrees
of inhibition against tested microbes. Overall, compounds
6e, 6f, 8a, 8b, 8d, and 8e showed the best antibacterial
activity with zone of inhibition ranging between 13.23
and 13.97 mm toward E coli. While the other compounds
showed less activity. Compounds 6f, 8b, 8c, and 8e pro-
duced good inhibitory potency against S aureus with the
inhibition zone >15 mm. Among these compounds 6f
(17.83 0.41 and 20.04 0.43 mm) in first series and 8b
(15.02 0.10 and 18.61 0.78 mm) in the second series
were more active against E coli and S aureus. Analysis of
the antibacterial results reveals that compounds 6e (-Br),
3.2 | Antifungal activity
Synthesized compounds 6a-f and 8a-e were also tested
for their in vitro antifungal activity using poisoned food
technique against C albicans and Aspergillus flavus. Com-
pound 8c with electron donating group CH3 at ortho
position of benzyl ring showed good inhibition with
inhitbition zone (24.17 0.32 &15.02 0.41 mm)
against C albicans and A flavus when compared to a stan-
dard antifungal drug, Nystatin. Whereas remaining com-
pounds showed less activity toward tested fungi. Results
of antifungal activity are given in the Table 2 and
Figure 2 (Graphically).
3.3 | Anthelmintic activity
Anthelmintic activity screening was done using
Pheretima posthuma (Indian earthworm) and the results
were given in the Table 3. Results revealed that, halogen
substituted at the para position of phenyl ring linked to
sulphonamide 6d showed good activity, paralysis starts at
37.45 0.27 minute and results in death at 40.03
1.23 minutes. This is may be due to Cl substituent a
key factor in pronouncing biological activity. Other
TABLE 1 Antibacterial activities of compounds (6a-f)
and (8a-e)
Inhibition zone (in mm)
(Mean SE)
Code
Control
6a
Conc (μg/10 μL) E. coli
S. aureus
–
–
–
10
10
10
10
10
10
10
10
10
10
10
10
12.02 0.76
11.89 0.63
10.21 0.72
12.83 0.23
13.97 0.31
17.83 0.41
13.56 0.62
15.02 0.10
12.07 0.55
13.89 0.22
13.23 0.62
20.00 0.50
11.67 0.25
11.19 0.25
11.01 0.17
9.86 0.41
12.97 0.32
20.04 0.43
11.37 0.10
18.61 0.78
16.13 0.30
11.84 0.16
15.89 0.15
25.00 0.50
6b
6c
6d
6e
6f
8a
8b
8c
8d
8e
CIPRO
FIGURE 1 Graphical representation of antibacterial activity
Note: Values are the mean SEM of the clear zone. Standard CIPRO,
Ciprofloxacin; Control, DMSO.
of compounds

4
KHAN ET AL.
TABLE 2 Antifungal activities of compounds 6(a-f) and 8(a-e)
TABLE 3 Anthelmintic activities of compounds 6(a-f) and
8(a-e)
Inhibition zone (in mm)
(Mean SE)
Conc
Time taken (min)
Treatment
Comp.
Control
6a
(μg/10 μL) C. albicans
A. flavus
code
(μg/μL)
Paralysis
Death
–
–
–
Control
6a
–
–
–
10
10
10
10
10
10
10
10
10
10
10
10
10.65 0.19
10.92 0.42
9.01 0.22
11.54 0.22
11.77 0.89
14.23 0.27
14.91 0.25
13.07 0.36
24.17 0.32
14.71 0.29
14.12 0.31
29.33 0.13
06.14 0.46
06.56 0.52
5.97 0.42
07.81 0.13
07.27 0.18
12.26 0.12
12.84 0.42
11.13 0.26
15.02 0.41
11.78 0.21
12.26 0.80
21.33 0.23
50
50
50
50
50
50
50
50
50
50
50
50
59.98 1.46
60.45 0.27
59.12 2.63
37.45 0.27
60.97 0.43
37.79 0.29
64.51 2.32
65.86 0.21
63.03 0.27
62.73 0.24
65.03 2.25
47.00 0.58
62.91 1.26
61.68 1.23
62.03 0.60
40.03 1.23
62.09 1.02
38.83 0.41
65.07 1.16
66.77 0.60
64.76 0.28
64.98 0.73
66.95 1.22
49.17 0.48
6b
6b
6c
6c
6d
6d
6e
6e
6f
6f
8a
8a
8b
8b
8c
8c
8d
8d
8e
8e
NYS
PC
Note: Values are the mean SEM of the clear zone. Standard NYS, Nystatin;
Note: Values are the mean SEM of three earthworms. Standard PC,
Control, DMSO.
Piperazine citrate. Control: Distilled water.
FIGURE 3 Graphical representation of antihelmintic activity
of compounds
FIGURE 2 Graphical representation of antifungal activity of
compounds
compound 6f with a nitro substituted at para position of
phenyl ring showed similar activity, where paralysis
starts at 37.79 0.29 minute and death of earth worm at
38.83 0.41 minute as shown in Figure 3. Compounds of
second series showed moderate activity against P post-
huma with respect to the reference drug, Piperazine Cit-
rate. The results of the anthelmintic activity are given
graphically in Figure 3.
500, 250, 125, and 62.5 μg/mL) against A375, MDA-MB
231 cancer cell line and normal HEK293 cells. The
CTC50 values for all tested compounds in the above cell
lines are shown in Table 4 and graphically in Figure 4.
Among all, compound 8a with dichloro substitution in
the benzyl ring showed higher toxicity of CTC50 of 22.18
1.3, 31.38 1.2 and 95.22 1.8 μg/mL, respectively
against A375 (melanoma cancer) and MDA-MB 231
(breast cancer) and normal HEK293 cell lines. While the
compound 8b showed good cytotoxic activity (CTC50
26.63 0.69, 39.12 1.5 μg/mL, respectively) on A375
and MDA-MB 231 cancer cell lines and no toxicity
(CTC50 140.22 0.15 μg/mL) against normal HEK293
cells. All other compounds showed less or moderate
3.4 | Cytotoxic activity
In vitro cytotoxic activity of compounds 6(a-f) and 8(a-e)
were evaluated at different concentrations (1000,

KHAN ET AL.
5
TABLE 4 Cytotoxicity of 6(a-f) and 8(a-e) against MDA–MB
231 A375 and HEK293 cell lines CTC50 in (μg/ml)
and fluoro substituent at the sixth position of benzyl ring
exhibited similar good specific cytotoxocity activity
toward cancerous cell lines A375 and MDA-MB 231 with
CTC50 value of 26.63 0.69 and 39.12 1.5 μg/mL,
respectively. Remaining compounds showed moderate
cytotoxic activity.
Code
6a
A375
MDA–MB 231
97.12 1.2
108.47 1.0
96.55 1.8
101.18 2.0
102.10 1.6
123.01 2.5
31.38 1.20
39.12 1.5
71.53 2.3
74.77 10
Normal HEK293
123.14 1.25
154.11 1.05
125.13 0.35
164.10 0.75
172.20 1.24
135.22 1.5
95.22 1.8
83.06 2.3
84.09 3.1
89.18 1.2
99.32 1.4
81.41 2.3
91.96 0.26
22.18 1.30
26.63 0.69
66.19 5.1
67.32 9.7
59.47 4.2
08.28 0.4
6b
6c
6d
6e
6f
3.5 | Molecular docking
8a
8b
140.22 0.15
142.23 1.32
186.32 1.56
125.25 1.7
39.63 1.35
The molecular docking studies were carried out using
Lead IT software. Docking energy of the synthesized
molecules along with standards were tabulated in
Table 5. All the molecules were docked effectively in
the active sites of respective target proteins. The Molec-
ular docking against 3ACX protein shows that com-
pounds 6f and 8b have good docking energy of −8.09
and − 7.733 kcal/mol respectively compared with
antibacterial standard ciprofloxacin. Compound 8c has
moderate docking score of −10.511 kcal/mol compared
to antifungal standard Nystatin. Docking energy of the
anthelmintic target shows that compounds 6d and 6f
with high docking score of −12.121 and − 12.511 kcal/
mol emerged as active compounds compared with stan-
dard Piperazine citrate whose docking energy is
−9.254 kcal/mol. Docking on B-Raf Kinase V600E onco-
genic mutant shows that compounds like 6f and 8a
show good docking energy. Since conventional hydro-
gen bonding is important, our molecular docking dis-
cussion is restricted to hydrogen bonding even though
compounds show interactions like Hydrophobic, Pi-Pi
stacked Pi-alkyl interactions.
8c
8d
8e
63.10 1.21
24.57 1.2
Cisplatin
FIGURE 4 Graphical representation of cytotoxicity activity of
compounds
activity when they are exposed to melanoma and breast
cancer cell lines. The graphical representation of the
cytotoxicity results of the compounds were given in
Figure 4.
3.6 | Binding mode of potent
antibacterial compounds
Two compounds 6f and 8b were involved in H-bonding
with the Asp48 of carotenoid dehydrosqualene synthase
with high docking score indicating that those are directly
involved in inhibiting the target protein. These two com-
pounds were involved in other interactions like van der
Waals and pi-pi. The fitting of these potent compounds
against bacteria protein 3ACX was shown in Figure 5.
1,2,4-Triazole, being an active pharmacophore (which
allows its derivatives to readily interact with the diversity
of enzymes and receptors in organisms) when linked
with aromatic ring substituted with electron donating
and electron withdrawing groups produced various
degrees of results. Our final compounds possessing these
substituents CH3, OCH3 (electron donating), F,
Cl, and NO2 (electron withdrawing) attached to ben-
zene moiety produced some good results. Compound 8a
with dichloro substituents at the third and fourth posi-
tion of benzyl ring showed good cytotoxicity effect
against cancerous A375, MDA-MB 231 and normal
HEK293 cell lines with CTC50 value of 22.18 1.30,
31.38 1.20, and 95.22 1.8 μg/mL respectively.
Whereas compound 8b with chloro substituent at second
3.7 | Binding mode of potent antifungal
compound
Compound 8c forms two hydrogen bonds with amino
acid residue Asp412 and also it shows some hydrophobic
interactions with other residues. The fitting of the potent
8c with fungi protein 1IYK was shown in Figure 6.

6
KHAN ET AL.
TABLE 5 Docking energy in kcal/
mol of target molecules
Antibacterial
Antifungal
Anthelmintic
Cytotoxic
Code
6a
PDBID: 3ACX
−7.701
−7.505
−7.665
−6.18
PDBID: 1IYK
−7.115
−8.212
−7.532
−7.176
−7.773
−7.306
−8.921
−9.121
−10.511
−6.248
−9.463
–
PDBID: 1SA0
−6.115
−6.712
−7.176
−12.121
−7.273
−12.511
−7.921
−7.532
−7.801
−10.248
−10.463
–
PDB ID: 3OG7
−11.251
−7.158
−11.172
−11.558
−9.164
−12.032
−13.897
−11.596
−8.271
−11.482
−11.558
–
6b
6c
6d
6e
−6.181
−8.09
−5.272
−7.733
−5.194
−5.712
−5.217
−9.18
6f
8a
8b
8c
8d
8e
Cipro
NYS
P C
–
−23.499
–
–
–
–
−9.254
–
FIGURE 5 Binding pattern of
compounds 6f and 8b with
protein 3ACX
3.8 | Binding mode of potent
anthelmintic compounds
group with COOH group of Thr145. Two hydrogen inter-
actions were observed between residue Gly146 and NH
group of sulfonamide linkage and one N1 atom of tri-
azole nucleus. There is another hydrogen bond
established between N2 atom of triazole nucleus with
Ser147. Glu183 residue accepting hydrogen from N4 atom
Molecular docking of the compound 6d and 6f into the
active site of target Tubulin shows each compound shows
hydrogen interaction between oxygen atom of its SO2

KHAN ET AL.
7
FIGURE 6 Binding pattern of
compounds 8c with protein 1IYK
FIGURE 7 Binding pattern of
compounds 6d and 6f with protein 1SA0
of triazole nucleus. These two compounds show similar
kind of interactions. The fitting of these compounds in
the pocket of protein 1SA0 are shown in Figure 7.
of Leu352. The binding pattern of this compound in the
active site of target protein PDB ID 3OG7 was shown in
Figure 8.
3.9 | Binding mode of potent cytotoxic
3.10 | Fingerprint analysis
compound
Fingerprints analysis has given more than a century of
credible proof for criminal inquiries due to their unique-
ness and persistence. Fingerprints that are commonly
used and invisible to the naked eye are Latent finger-
prints (LFPs). Extraordinary fluorescent powders were
Molecular docking of the compound 8a into the active
site of cancer protein reveals that there is one conven-
tional hydrogen bond formed with Leu352 by the dona-
tion of hydrogen from triazole nucleus to carboxyl group

8
KHAN ET AL.
FIGURE 8 Binding pattern of
compound 8a with protein PDB
ID 3OG7
FIGURE 9 Fingerprint
visualization of samples 6(a-d) and 8(a-
d) on the surface of aluminum foil under
normal light
used to improve LFPs in crime scene. Rare-earth
materials,[25] carbon dots,[26] nanoparticles and
nanotubes,[27,28] Tetraphene derivatives,[29] 1,4-disub
stituted 1,2,3-triazoles,[30] and so forth, have been
reported for imaging of LFPs. The powder used in this
technique still has many problems, such as high back-
ground interference, low contrast, high autofluorescence
interactions, and low sensitivity.[31,32] Therefore, it is very
essential to develop novel materials to accomplish simple
and fast LFPs imaging. Synthesized compounds 6(a-d)
and 8(a-d) were subjected to eccrine LFPs development
on the surface of aluminum foil to determine their label-
ing efficacy in the forensic analysis are shown in
Figure 9.
Sample 6d shows the clear ridge details among
all samples, with enhanced ridge is further explored
to visualize the fingerprint on various nonporous sur-
faces such as TV remote, glass, metal scale, goggle,
compact disc, and marble under normal light as
shown in the Figure 10. The obtained results clearly
show high sensitivity of the prepared samples for
LFPs imaging.

KHAN ET AL.
9
FIGURE 10 Fingerprint visualization of sample 6d on nonporous surfaces, A and B, magazine cover, c, glass slide, D, spray bottle
cap, E, compact disc, F, black plastic bottle, G, coin, H, granite, I, spoon, and, j, heighlater pen surface using optimized sample under normal
light
4 | CONCLUSION
4.1 | Experimental section
In this article, a series of novel compounds were
synthesized and screened for their antibacterial, anti-
fungal, anthelmintic, and cytotoxic activities. Out of
all tested compounds, 6f in the sulfonamide series
and 8b in the second series showed relatively good
antibacterial activity against S aureus and E coli
and 8c emerged as a prominent antifungal agent
against C albicans and A flavus, while the com-
pounds 6d and 6f in the first series exhibited
remarkable anthelmintic activity. Compound 8a
emerged as potent anticancer agent against A375
(melanoma cancer) and MDA-MB 231 (breast can-
cer) cell lines. Docking studies were also performed
for all synthesized compounds in order to examine
the binding pattern of active molecules. Fingerprint
application study reveals that compound 6d has
good adhesive property which is useful for finger-
print applications.
All solvents and chemicals were bought from Sigma-Aldrich
and Spectrochem in appropriate grade. Open capillaries
were used to assess the melting points of the compounds.
Pre-coated TLC plates were used to ascertain the purity of
compounds and solvent system used in this experiment was
Ethyl acetate/Petroleum ether (3:7). Column chromatogra-
phy was carried out using Merck silica gel 60. UV light
(254 nm) or iodine chamber visualization was performed.
4.2 | Chemistry
4.2.1 | Procedure for the synthesis of
ethyl 2-amino-5-bromobenzohydrazide (3)
Ethyl 2-amino-5-bromobenzohydrazide (3) was prepared
from ethyl 2-amino-5-bromobenzoate according to the lit-
erature method.[33]

10
KHAN ET AL.
4.2.2 | Procedure for the synthesis
4.3 | Spectral characterization
4-bromo-2-(4H-1,2,4-triazole-3-yl)aniline (4)
N-(4-Bromo-2-[4H-1,2,4-triazol-3-yl]phenyl)-
4-methylbenzenesulfonamide (6a).
A mixture of ethyl 2-amino-5-bromobenzohydrazide (3)
(0.1 mol), formamide (0.15 mol) in DMF were refluxed at
200ꢀC in presence of pulverized potassium hydroxide for
about 3 hours. Progress of the reaction was observed by
TLC. After completion of the reaction, the reaction mix-
ture was diluted with excess of water to remove DMF,
then compound was extracted with ethylacetate
(50 mL × 3). The ethylacetate layer was washed with
water and brine. The final organic layer was dried over
anhydrous Na2SO4. Then ethylacetate was distilled off
under reduced pressure to get compound (4) as a yellow
powder. The product was recrystallized from methanol
and dried. It was confirmed by 1H-NMR (400 MHz,
DMSO-d6) δ ppm: 6.68 (s, 2H, N H), 6.74 to 6.76 (d,
J = 8 Hz, 1H, Ar H), 7.20 to 7.22 (d, J = 8 Hz, 1H,
Ar H), 7.99 (s, 1H, Ar H), 8.49 (s, 1H, Ar H), 14.2 (s,
1H, triazole-N H); Mass: cal 238; found m/z: 239 [M
+ 1] and 241 [M + 2].
Color: yellow powder; yield: 91%; m.p: 154ꢀC. IR
(KBr, ν cm−1): 3462 (N-H), 3360 (H NSO2), 3131
(Ar H), 2921 (C H), 1389 (S O asymmetric), 1194
(S O symmetric). 1H-NMR (400 MHz, DMSO-d6), δ,
ppm: 2.37 (s, 3H, CH3), 6.2-6.8 (b, s, 1H, H NSO2), 6.9
(s, 1H, Ar H) 6.74-6.76 (d, J = 8, 1H, Ar H), 7.25 to 7.26
(d, J = 4 Hz, 1H, Ar H), 7.4 to 7.5 (d, J = 4, 2H, Ar H),
7.92 (s, 1H, Ar H), 8.01 to 8.03 (d, J = 8, 2H, Ar H),
9.45 (s, 1H, triazole-N H). 13C-NMR (100 MHz, DMSO-
d6), δ, ppm: 21.68, 106.1, 111.6, 118.6, 128.77, 130.6,
131.17, 132.5, 134.1, 146.7, 146.8, 147.8, 163.4. Mass: m/z
calculated for C14H11BrN4O2S: 391.99, Found: 393.0 [M
+ 1] and 395.0 [M + 2].
N-(4-Bromo-2-[4H-1,2,4-triazol-3-yl]phenyl)-
4-methoxybenzene sulfonamide (6b). Color: light yellow
powder; m.p: 180ꢀC; yield: 75%. IR (KBr, ν cm−1): 3450
(N H), 3356 (H-NSO2), 3137 (Ar H), 2971 (C H),
1382.2 (S O asymmetric), 1194 (S O symmetric).
1H NMR (400 MHz, DMSO-d6), δ, ppm: 3.83 (s, 3H,
OCH3), 5.5-6.7 (b, s, 1H, H NSO2), 6.8 (s, 1H, Ar H),
6.74 to 6.77 (d, J = 12 Hz, 1H, Ar H), 7.17 to 7.19 (d,
J = 8 Hz, 2H, Ar H), 7.24 (S, 1H, Ar H), 7.92 to 7.93 (d,
J = 4 Hz, 1H, Ar H), 8.06 to 8.086 (d, J = 8 Hz, 2H,
Ar H), 9.42 (s, 1H, triazole-N H). 13C NMR (100 MHz,
DMSO-d6), δ, ppm: 56.56, 106.24, 111.89, 118.7, 115.9,
126.3, 130.64, 131.4, 134.02, 146.6, 146.7, 163.3, 165.4.
Mass: m/z calculated for C15H13BrN4O3S: 407.99, Found:
409.0 [M + 1] and 411.0 [M + 2].
4.2.3 | General procedure for the
preparation of compounds 6(a-f)
Equimolar quantities of 4-bromo-2-(1H-1,2,4-triazol-5-yl)
aniline (4) (0.001 mol), different substituted benzene sul-
fonyl chlorides 5(a-f) (0.001 mol) and triethylamine were
taken in methylene dichloride. Reaction mixture was
stirred for 30 minutes at room temperature till solid sepa-
rates out and completion of the reaction was checked by
TLC. Obtained solid product was separated by filtration.
Then it was dried and purified by recrystallization using
20% ethanol to get 4-bromo-bezenesulfonamide deriva-
tives bearing 1,2,4-triazole in high yield 6(a-f).
5-Bromo-N-(4-bromo-2-[4H-1,2,4-triazol-3-yl]phenyl)
thiophene-2-sulfonamide (6c). Color: light yellow powder;
m.p: 196ꢀC; yield: 79%. IR (KBr, ν cm−1): 3469 (N H),
3368 (H-NSO2), 2923.5 (Ar H), 1390 (S O asymmetric),
1
1121 (S O symmetric). H-NMR (400 MHz, DMSO-d6),
δ, ppm: 6.53 (s, 1H, H-NSO2), 6.77 (s, 1H, Ar H), 6.79 (s,
1H, Ar H), 7.26-7.27 (d, J = 4 Hz, 1H, Ar H), 7.51 to
7.52 (d, J = 4 Hz, 1H, Ar H), 7.95 to 7.96 (d, J = 4 Hz,
1H, Ar H), 8.05 to 8.06 (d, J = 8 Hz, 1H, Ar-H), 9.46 (s,
1H, triazole-N-H). 13C NMR (100 MHz, DMSO-d6) δ
ppm: 106.21, 118.7, 111.5, 126.3130.7, 133.3, 134.2, 134.7,
139.2, 146.84146.96163.5. Mass: m/z calculated for
C12H8Br2N4O2S2: 461.85, Found: 462.88 [M + 1] and
464.88 [M + 2].
4.2.4 | General procedure for the
preparation of compounds 8(a-e)
Equimolar quantities of 4-bromo-2-(1H-1,2,4-triazol-5-yl)
aniline, different substituted benzyl bromide 5(a-e)
(0.001 mol) in acetonitrile were stirred for 10 hours at
30ꢀC in presence of K2CO3 (0.003 mol). The completion
of the reaction was checked by TLC. After the comple-
tion, it was quenched by pouring onto ice-cold water then
the product was extracted with ethyl acetate. After drying
over anhydrous sodium sulfate, ethylacetate was concen-
trated under reduced pressure to get alkylated
1,2,4-triazole derivative 8(a-e) as solids in good yield.
Compounds 8(a-e) were purified by recrystallization
using ethylacetate: n-hexane (9:1).
N-(4-Bromo-2-[4H-1,2,4-triazol-3-yl]phenyl)-
4-chlorobenzenesulfonamide (6d). Color: light yellow pow-
der; m.p: 186ꢀC; yield: 77%. IR (KBr, ν cm−1): 3465
(N H), 3364 (H-NSO2), 2923.6 (Ar H), 1393 (S O asym-
metric), 1186 (S O symmetric). 1H NMR (400 MHz,
DMSO-d6), δ, ppm: 5.1 to 6.7 (b, s, 1H, H-NSO2), 7.25 to
7.28 (d d, J = 4 Hz, 2H, Ar H), 7.77 to 7.79 (d, J = 8 Hz,

KHAN ET AL.
11
2H, Ar H), 7.93 (s, 1H, Ar H), 7.57 (s, 1H, Ar H), 8.15
to 8.17 (d, J = 8 Hz, 2H, Ar H), 9.48 (s, 1H, triazole-
N H).13C NMR (100 MHz, DMSO-d6), δ, ppm: 106.32,
118.8, 127.9, 128.0, 129.9, 130.2, 132.8, 141.8, 145.7, 146.7,
147.15, 163.6. Mass: m/z calculated for C14H10BrClN4O2S:
411.94, Found: [M + 1] and 414.97 [M + 2].
4-Bromo-N-(4-bromo-2-[4H-1,2,4-triazol-3-yl]phenyl)-
4-benzenesulfonamide (6e). Color: Brown solid; m.p:
142ꢀC to 145ꢀC; yield: 90%. IR (KBr, ν cm−1): 3437
(N H), 3312 (H NSO2), 2901 (Ar H), 1343.2 (S O
asymmetric), 1181.5 (S O symmetric). 1H NMR
(400 MHz, DMSO-d6), δ, ppm: 5.6-5.7 (b, s 1H, H NSO2),
6.81 to 6.83 (d, J = 8 Hz, 2H, Ar H), 7.9 (s, 1H, Ar H),
7.91 (s, 1H, Ar H), 8.06 to 8.08 (d, J = 8 Hz, 1H, Ar H),
7.283 to 7.289 (d, J = 2.4 Hz, 2H, Ar H), 8.61 (s, 1H,
Ar H), 9.48 (s, 1H, triazole-N H). 13C NMR (100 MHz,
DMSO-d6), δ, ppm: 119.82, 122.07, 128.2, 129.1, 130.3,
131.05, 132.9, 133.35, 145.6, 143.7, 148.06, 157.6. Mass:
m/z calculated for C14H10Br2N4O2S: 455.89; Found:
457 [M + 1] and 459 [M + 2].
(400 MHz, DMSO-d6), δ, ppm: 5.58 (s, 2H, CH2), 6.48
(b, s, 1H, N H), 6.70 (s, 1H, Ar H), 7.16 to 7.18 (m, 1H,
Ar H), 7.183 to 7.188 (d, J = 2 Hz, 1H, Ar H), 7.30 to
7.32 (d, J = 8 Hz, 1H, Ar H), 7.37 to 7.39 (d, J = 8 Hz,
1H, Ar H), 7.48 (s, 1H, Ar H), 7.91 to 7.92 (d, J = 4 Hz,
1H, Ar H), 8.76 (s, 1H, triazole-N H).13C NMR
(100 MHz, DMSO-d6), δ, ppm: 44.3, 106.1, 114.06, 115.4,
118.2, 121.5, 126.2, 129.9, 132.1, 135.4, 145.1146.07, 160.6,
161.08, 163.09. Mass: m/z calculated for C15H11BrClFN4:
379.98, Found: 380.9 [M + 1] and 382.9 [M + 2].
4-Bromo-N-(2-methylbenzyl)-2-(4H-1,2,4-triazol-3-yl)
aniline (8c). Color: light yellow powder; m.p: 119ꢀC;
yield: 91%. IR (KBr, ν cm−1): 3420 (N H), 3298
(Ar NH), 3078 (Ar H), 2913 (C H). 1H NMR
(400 MHz, DMSO-d6), δ, ppm: 2.34 (s, 3H, CH3), 5.47 (s,
2H, CH2), 6.2-6.6 (b, s, 1H, N H), 6.73 to 6.75 (d,
J = 8 Hz, 1H, Ar H), 7.07 to 7.08 (d, J = 7.6 Hz, 1H,
Ar H), 7.14 (s, 1H, Ar H), 7.16 to 7.20 (m, 4H, Ar H),
7.99 (s, 1H, Ar H), 8.7 (s, 1H, triazole-N H). 13C NMR
(100 MHz, DMSO-d6), δ, ppm: 19.15, 50.94, 106.22,
114.47, 118.32, 126.67, 128.60, 128.94, 130.03, 130.81,
132.42, 134.61, 136.58, 144.91, 146.11, 161.03. Mass: m/z
calculated for C16H15BrN4: 342.05, Found: 343.0 [M + 1]
and 345.0 [M + 2].
4-Bromo-N-(4-fluorobenzyl)-2-(4H-1,2,4-triazol-3-yl)
aniline (8d). Color: light yellow powder; m.p: 168ꢀC;
yield: 92%. IR (KBr, ν cm−1): 3305 (N H), 3274
(Ar NH), 1690.2 (C C, Ar). 1H NMR (400 MHz,
DMSO-d6), δ, ppm: 4.41 (s, 1H, N H), 5.44 (s, 2H,
CH2), 6.62 to 6.64 (d, 1H, J = 8 Hz, Ar H), 6.75 to 6.77
(d, 1H, J = 8 Hz, Ar H), 7.10 to 7.25 (m, 4H, Ar H),
7.20 (s, 1H, Ar H), 7.9 (s, 1H, Ar H), 8.8 (s, 1H,
triazole-N H). 13C NMR (100 MHz, DMSO-d6), δ, ppm:
46.14, 106.67, 113.9, 114.7, 115.58, 116.08, 118.64, 129.5,
130.7, 132.70, 135.6, 144.7, 145.6. Mass: m/z calculated
for C15H12BrFN4: 346.02, Found: 347 [M + 1] and
349 [M + 2].
4-Bromo-N-(4-methylbenzyl)-2-(4H-1,2,4-triazol-3-yl)
aniline (8e). Color: Dark yellow powder, m.p: 159ꢀC;
yield: 90%. IR (KBr, ν cm−1): 3310 (N H), 3274
(Ar NH), 3109 (Ar H), 2942 (C H). 1H-NMR
(400 MHz, DMSO-d6), δ, ppm: 2.4 (s, 3H, CH3), 5.3 (s,
2H, CH2), 6.20 to 5.91 (b, s, 1H, N H), 6.34 to 6.36 (d,
J = 8 Hz, 1H, Ar H), 6.92 (s, 1H, Ar H), 7.12 to 7.13 (d,
J = 4 Hz, 2H, Ar H), 7.14 to 7.16 (d, J = 8 Hz, 1H,
Ar H), 7.40 to 7.60 (d, J = 8 Hz, 2H, Ar H), 7.99 (s, 1H,
Ar H) 8.9 (s, 1H, triazole-N H). 13C NMR (100 MHz,
DMSO-d6), δ, ppm: 18.68, 51.24, 105.62, 113.47, 119.02,
125.23, 127.35, 129.60, 131.05, 133.33, 135.24, 143.24,
145.21, 162.12. Mass: m/z calculated for C16H15BrN4:
342.05; Found: 343.0 [M + 1] and 345.0 [M + 2].
N-(4-Bromo-2-[4H-1,2,4-triazol-3-yl]phenyl)-
4-nitrobenzenesulfonamide (6f). Color: White solid; m.p:
178ꢀC; yield: 82%. IR (KBr, ν cm−1): 3441 (N H), 3291
(H NSO2), 2971 (Ar H), 1323.8 (S O asymmetric),
1
1165.2 (S O symmetric). H-NMR (400 MHz, DMSO-d6)
δ ppm: 6.66 (b, s, 1H, H NSO2), 7.27 to 7.28 (d, J = 4 Hz,
2H, Ar H), 7.61 (s, 1H, Ar H), 8.02 to 8.04 (d, J = 8 Hz,
2H, Ar H), 8.22 (s, 1H, Ar H), 8.41 to 8.40 (d, J = 4 Hz,
2H, Ar H), 8.80 (s, 1H, triazole-N H). 13C NMR
(100 MHz, DMSO-d6), δ, ppm: 119.82, 121.9, 122.3, 124.6,
129.27, 131.9, 132.17, 133.19, 142.3, 144.2, 155.70, 156.9.
Mass: m/z calculated for C14H10BrN5O4S: 422.96, Found:
424 [M + 1] and 426 [M + 1].
4-Bromo-N-(3.4-dichlorobenzyl)-2-(4H-1,2,4-triazol-
3-yl)aniline (8a). Color: light yellow powder; m.p: 138ꢀC;
yield: 91%. IR (KBr, ν cm−1): 3433 (N H), 3334
(Ar NH), 3090 (Ar H), 2926 (C H). 1H-NMR
(400 MHz, DMSO-d6), δ, ppm: 5.48 (s, 2H, CH2), 5.8 to
6.40 (b, s, 1H, N H), 6.74 (s, 1H, Ar H), 7.178 to 7.185
(d, J = 2.8 Hz, 1H, Ar H), 7.29 (s, 1H, Ar H), 7.31 to
7.32 (d, J = 4 Hz, 1H, Ar H), 7.60 to 7.65 (d d, J = 1.6 Hz,
8 Hz, 2H, Ar H), 7.96 (s, 1H, Ar H), 8.79 (s, 1H,
triazole-N H). 13C NMR (100 MHz, DMSO-d6), δ, ppm:
51.4, 106.28, 114.2, 118.41, 128.81, 130.02, 130.5, 131.26,
131.3, 131.7, 132.5, 137.4, 145.09, 146.00, 161.3. Mass: m/z
calculated for C15H11BrCl2N4: 395.95, Found: 396.8 [M
+ 1] and 398.87 [M + 2].
4-Bromo-N-(2-chloro-6-fluorobenzyl)-2-(4H-1,-
2,4-triazol-3-yl)aniline (8b). Color: light yellow powder;
m.p: 169ꢀC; yield: 90%. IR (KBr, ν cm−1): 3441 (N H),
3342 (Ar NH), 3082 (Ar H), 2958 (C H). 1H NMR

12
KHAN ET AL.
4.4 | Biological screening
cells using MTT assay.[39,40] Cell lines were obtained
from National Centre For Cell Science (NCCS Pune,
India), cultured in Dulbecco's modified Eagle's medium
(DMEM) supplemented with 10% heat-inactivated fetal
bovine serum, 2 mmoL/L glutamine, 1% antibiotics
solution (100 U/mL penicillin G and 100 mg/mL strep-
tomycin). Cells were grown in 25 cm2 tissue culture
flasks and maintained at 37ꢀC in a humidified incuba-
tor with 5% CO2. Cytotoxic activity results were
expressed in the form of CTC50 (Concentration of test
drug needed to inhibit cell growth by 50%) and Cis-
platin was used as a standard drug. All the series of
compounds were screened for their cytotoxicity activity
against three different cell lines at different concentra-
tions of 1000, 500, 250, 125, and 62.5 μg/mL in
dimethylsulfoxide (DMSO).
4.4.1 | Antibacterial activity
Antibacterial activity of synthesized compounds were
assessed by Disc-diffusion method[34–36] against two bacte-
rial strain viz S aureus and E coli. Bacterial strains were
maintained on Muller-Hinton agar medium. Ciprofloxacin
was used as reference antibacterial drug. Agar plates were
inoculated with inoculum of S aureus and E coli. Then fil-
ter paper discs of about 6 mm diameter containing differ-
ent concentrations of 2.5, 5, 10, 15, and 20 μg in dimethyl
sulfoxide were placed on the agar medium. The petri dis-
hes were incubated at 37ꢀC. After incubation, the diameter
of the zone of inhibition (ZOI) was measured in mm. Most
of the compounds showed ZOI at 10 μg/10 μL as Mini-
mum inhibitory concentration (MIC).
4.5 | Statistical analysis
4.4.2 | Antifungal activity
Cytotoxicity was assessed using the CTC50 value (50%
cell growth inhibition) as a response parameter, which
was calculated with the GraphPad Prism 8.0 program.
The poisoned food technique method was used against C
albicans and A flavus to assess the antifungal activity of
the synthesized compounds.[37] Different concentrations
of 2.5, 5, 10, 15, and 20 μg in dimethyl sulfoxide was
incorporated into the molten agar and mixed well. The
medium was poured into petri dishes and kept for incu-
bation at 37ꢀC overnight. Inoculation with fungal strains
was done and petri plates were further incubated at 37ꢀC.
After incubation zone of inhibition was measured in
mm. 10 μg/10 μL was noticed as a MIC since all com-
pounds showed inhibition at this concentration. Nystatin
was the standard antifungal drug used.
4.6 | Molecular docking
Structure of synthesized molecules was drawn by using
Chem Draw.[41,42] Energy minimization protocol and
macro molecular force field was used to perform Ligand
optimization.[39] Different ligand conformations were
produced on the basis of their dihedral energy, bond
energy, initial potential energy, CHARM energy values.
Using the Lipinski rule of five via Lipinski drug filter
protocol, drug likeliness was determined.[43,44] Discov-
ery studio 3.5 (Accelrys) was used to carried out these
studies.
4.4.3 | Anthelmintic activity
Anthelmintic activity of the compounds was evaluated
using P posthuma (Indian Earthworm) using standard
method.[38] Test samples of compounds at 10, 20, and
50 mg/mL in distilled water and P posthuma were placed
in each Petri dish containing 25 mL of test solution.
Piperazine citrate was used as standard and distilled
water as control. Observations were made for the time
taken for paralysis was noted when no movement was
observed. Time for death of worms were recorded after
confirming that worms neither moved when shaken vig-
orously nor when dipped in warm water (50ꢀC).
4.7 | Fingerprint application
Synthesized compounds 6(a-f) and 8(a-e) were exam-
ined for their labeling efficacy in forensic applications
on aluminum foil surface. All eccrine LFPs were col-
lected by a donor using cleaned and dried hands. Later,
the fingers were pressed at low pressure on various sur-
faces with infiltrating and noninfiltrating materials at
37ꢀC temperature to obtain LFP. Each compound is
carefully applied to the eccrine LFPs in order to
improve the LFP and the excess compound powder was
eliminated by soft brushing method. The obtained fin-
gerprints are photographed with normal white light
with a digital camera Nikon D3100/AF_S Nikkor50 mm
f/2.8G ED.[45]
4.4.4 | Cytotoxic activity
Cytotoxic activity of all compounds was studied against
A375, MDA-MB 231 cancer cells and HEK293 normal

KHAN ET AL.
13
[20] I. Kuçukguzel, S. G. Kuçukguzel, S. Rollas II., Farmaco 2004,
59(11), 893.
[21] J. J. Tomasz Plech, M. W. Luszczki, J. Flieger, M. Pizon, Eur.
J. Med. Chem. 2013, 60, 208.
[22] M. M. Ghorab, M. S. Alsaid, M. Al-Dosari, Acta Pol. Pharm.
2016, 73(5), 1147.
ACKNOWLEDGMENTS
Authors thank the Tumkur University and Siddaganga
Institute of Technology for laboratory facilities. Authors
also thank JSS College of Pharmacy, Mysuru for biologi-
cal activity studies.
[23] D.
Chowrasiaa,
C.
Karthikeyana,
L.
Chourea,
ORCID
M. G. Sahabjadab, M. Arshadb, P. Trivedia, Arab. J. Chem.
2017, 10, S2424.
Swamy Sreenivasa https://orcid.org/0000-0003-1096-
[24] F. Gao, T. Wang, J. Xiao, G. Huang, Eur. J. Med. Chem. 2019,
173, 274.
5837
[25] M. Wang, M. Li, A.-Y. Yu, J. Wu, C.-B. Mao, ACS Appl. Mater.
Interfaces 2015, 7, 28110.
[26] J. Chen, J.-S. Wei, P. Zhang, X.-Q. Niu, W. Zhao, Z.-Y. Zhu,
H. Ding, H.-M. Xiong, ACS Appl. Mater. Interfaces 2017, 9,
18429.
REFERENCES AND NOTES
[1] D. Marchaim, F. Perez, J. Lee, S. Bheemreddy, A. M. Hujer,
S. Rudin, K. Hayakawa, P. R. Lephart, C. Blunden, M. Shango,
M. L. Campbell, J. Varkey, P. Manickam, D. Patel,
J. M. Pogue, T. Chopra, E. T. Martin, S. Dhar, R. A. Bonomo,
K. S. Kaye, Am. J. Infect. Control 2012, 40(9), 830.
[2] Y. J. Mange, A. M. Isloor, S. Malladi, S. Isloor, H. K. Fun,
Arab. J. Chem. 2013, 6, 177.
[3] S. Pattan, P. Gadhave, V. Tambe, S. Dengale, D. Thakur,
S. V. Hiremath, R. V. Shete, P. Deotarse, Indian J. Chem. 2012,
51B, 297.
[4] O. Bekircan, B. Kahveci, M. Kucuk, Turk. J. Chem. 2006,
30, 29.
[5] V. K. Kamboj, P. K. Verma, A. Dhanda, S. Ranjan, Med. Chem.
2015, 15, 17.
[6] X. Li, X. Q. Li, H. M. Liu, X. Z. Zhou, Z. H. Shao, Org Med
Chem Lett 2012, 2, 26.
[27] J. Wang, T. Wei, X.-Y. Li, B.-H. Zhang, J.-X. Wang, C. Huang,
Q. Yuan, Angew. Chem. Int. Ed. 2014, 53, 1616.
[28] M. Wang, L. Guo, D. Cao, Sci. China Chem. 2017, 60, 1090.
[29] H. Huang, Y. Zhou, M. Wang, J. Zhang, X. Cao, S. Wang,
D. Cao, C. Cui, Angew. Chem. Int. Ed. 2019, 58, 10132.
[30] S. Govindaiah, S. Sreenivasa, R. A. Ramakrishna, T. M. Chakrapani
Rao, H. Nagabhushana, Chem. Select. 2018, 3, 8111.
[31] C. Suresh, H. Nagabhushana, R. B. Basavaraj, G. P. Darshan,
D. B. Kavyashree, S. C. Daruka Prasad, R. Sharma,
J. Vanithamani, Colloid Interface Sci. 2018, 518, 200.
[32] K. N. Venkatachalaiah, H. Nagabhushana, G. P. Darshan,
R. B. Basavaraj, B. Daruka Prasad, Sensors Actuators B 2017,
251, 310.
[7] A. Cetin, I. H. Gecibesler, J Appl Pharm Sci. 2015, 5, 120.
[8] C. H. Zhou, Y. Wang, Curr. Med. Chem. 2012, 19, 239.
[9] A. Mojaddami, A. Sakhteman, M. Fereidoonnezhad,
Z. Faghih, A. Najdian, S. Khabnadideh, H. Sadeghpour,
Z. Rezaei, Res Pharm Sci. 2017, 12, 21.
[10] A. Krajczyk, K. Kulinska, T. Kulinski, B. L. Hurst, C. W. Day,
D. F. Smee, T. Ostrowski, P. Januszczyk, J. Zeidler, Antivir.
Chem. Chemother. 2014, 23, 161.
[11] V. Sumangala, B. Poojary, N. Chidananda, T. Arulmoli,
S. Shenoy, Eur. J. Med. Chem. 2012, 54, 59.
[12] D.-J. Fua, Y.-C. Liua, J.-J. Yanga, Z. Ji, C.-D. Xionga, Z.-
S. Caoa, X.-X. Yina, W. Weia, Y.-B. Zhanga, J. Chem. Res.
2017, 41, 523.
[13] M. G. El-Gazzar, N. H. Nafie, A. Nocentini, M. M. Ghorab,
H. I. Heiba, C. T. Supuran, J. Enzyme Inhib. Med. Chem. 2018,
33(1), 1565.
[14] S. R. Desai, U. Laddi, R. S. Bennur, P. A. Patil, S. Bennur, Ind.
J.Pharm. Sci. 2011, 73, 115.
[15] T. Okazaki, A. Suga, T. Watanabe, K. Kikuchi, H. Kurihara,
M. Shibasaki, A. Fujimori, O. Inagaki, U. Yanagisawa, Chem.
Pharm. Bull. 1998, 46, 69.
[16] R. Kharb, P. C. Sharma, M. S. Yar, J. Enzyme Inhib. Med.
Chem. 2010, 26(1), 1.
[33] X. Zheng, Z. Li, Y. Wang, W. Chen, J. Fluor. Chem. 2003,
123, 163.
[34] H. W. Seeley, P. J. Van Denmark, Laboratory manual of Micro-
biology, 2nd ed., Taraporewala D. B. Sons and Co, Microbes in
action, Bombay 1975, p. 55.
[35] N. R. Mohan, S. Sreenivasa, K. E. Manojkumar, T. M. C. Rao,
B. S. Thippeswamy, P. A. Suchetan, J. Braz. Chem. Soc. 2014,
25, 1012.
[36] N. R. Mohan, S. Sreenivasa, K. E. Manoj Kumar, H. C. Anitha,
T. Madhu Chakrapani Rao, B. S. Thippeswamy, V. K. Gowda,
Indo Am. J. Pharm. Res. 2013, 3, 1513.
[37] M. S. Balouiri, M. adiki, S. K. Ibnsouda, J. Pharma. Anal. 2016,
6, 71.
[38] B. Satish, Kosalge, A. Ravindra, Fursule. Asian J. Pharm. Clin.
Res. 2009, 2, 69.
[39] R. Anantacharya, N. D. Satyanarayan, B. S. Kalal, V. R. Pai,
Open Med. Chem. J. 2018, 12, 60.
[40] N. Kumar, S. Sreenivasa, K.S. Bhuvanesh, V. Kumar, B.
S. Holla, V. R. Pai, N. R. Mohan, G. Shivaraj, Lett. Drug Des.
Discovery,
2019,
16,
994,
Benzo[d]imidazol-5-yl)-
5-(substituted)-1,3,4-Oxadiazoles: Synthesis, Anticancer, Anti-
microbial and In Silico Studies, 1005.
[41] K. Wingen, J. S. Schwed, K. Isensee, L. Weizel, A. Ivkovi,
D. Odazic, H. Stark, Bioorg. Med. Chem. Lett. 2014, 24, 2236.
[42] Ghouse Khan, S. Sreenivasa, G. Shivaraja, V. Chandramohan,
Orient. J. Chem. 2019, 35, 157.
[43] G. Wu, D. H. Robertson, C. L. Brooks III., M. Vieth, J. Comput.
Chem. 2003, 24, 1549.
[17] J. Liu, Q. Liu, Y. Xue, S. Xu, H. Zhang, R. Bai, H. Yao, J. Jiang,
M. Shen, X. Wua, J. Xu, Med. Chem. 2013, 21, 7742.
[18] M. V. Papadopoulou, W. D. Bloomer, H. S. Rosenzweig,
E. Chatelain, M. Kaiser, S. R. Wilkinson, C. McKenzie, J.-
R. Ioset, J. Med. Chem. 2012, 55, 5554.
[19] K. S. Song, S.-H. Lee, H. J. Chun, J. Y. Kim, M. E. Jung, Bioorg.
Med. Chem. Lett. 2008, 16, 4035.
[44] C. A. Lipinski, Revolut. Drug Discov. Today Technol. 2004,
1, 337.

14
KHAN ET AL.
[45] B. S. Rohini, G. P. Darshan, H. B. Premkumar, D. Kavyashree,
S. C. Sharma, S. Sreenivasa, H. Nagabhushana, Colloids Surf.
A Physicochem. Eng. Asp. 2019, 123749, 581.
How to cite this article: Khan G, Sreenivasa S,
Govindaiah S, Chandramohan V, Shetty P R.
Synthesis, biological screening, in silico study and
fingerprint applications of novel 1, 2, 4-triazole
derivatives. J Heterocyclic Chem. 2020;1–14. https://
doi.org/10.1002/jhet.3929
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