Synthesis, antibacterial and antifungal activity of some novel 3,5-disubstituted-1H-1,2,4-triazoles

Article abstract of DOI:10.1002/ardp.200800005

A rapid and efficient one-pot condensation reaction of long-chain alkyl and alkenyl acid hydrazides and nitriles was carried out to afford 3,5-disubstituted-1H-1,2,4-triazoles. The compounds 5a-o were screened for in-vitro antibacterial activity against the representative panel of two Gram-positive and two Gram-negative bacteria. All the synthesized compounds were also tested for their inhibitory action against five strains of fungi. The various compounds show potent inhibitory action against test organisms. The compounds 5a-o were characterized on the basis of elemental analysis and spectral data.

Full text of DOI:10.1002/ardp.200800005

714
Arch. Pharm. Chem. Life Sci. 2008, 341, 714–720
Full Paper
Synthesis, Antibacterial and Antifungal Activity of Some Novel
3,5-Disubstituted-1H-1,2,4-triazoles
Shweta Sharma1, Saloni Gangal1, Abdul Rauf1, and Maryam Zahin2
1 Department of Chemistry, Aligarh Muslim University, Aligarh, India
2 Department of Agricultural Microbiology, Aligarh Muslim University, Aligarh, India
A rapid and efficient one-pot condensation reaction of long-chain alkyl and alkenyl acid hydra-
zides and nitriles was carried out to afford 3,5-disubstituted-1H-1,2,4-triazoles. The compounds
5a–o were screened for in-vitro antibacterial activity against the representative panel of two
Gram-positive and two Gram-negative bacteria. All the synthesized compounds were also tested
for their inhibitory action against five strains of fungi. The various compounds show potent
inhibitory action against test organisms. The compounds 5a–o were characterized on the basis
of elemental analysis and spectral data.
Keywords: 3,5-Disubstituted-1H-1,2,4-triazoles / Long-chain alkyl and alkenyl acid hydrazides / Nitriles /
Received: January 7, 2008; accepted: April 28, 2008
DOI 10.1002/ardp.200800005
Owing to its broad spectrum of biological activity [15–
Introduction
21], the 1,2,4-triazole ring system represents an attractive
target to invent new substrates for their synthesis and
production of combinatorial libraries. In several pharma-
cologically active compounds 3,5-disubstituted-1,2,4-tria-
zoles are found. Recent examples include selective adeno-
sine A2A receptor antagonist [22] and the phosphodiester-
ase V inhibitor [23]. Previously, 1,2,4-triazoles were syn-
thesized by hydrazides and nitriles either by Pinner reac-
tion and Pellizzari condensation which involve the cyclo-
dehydrative condensation between nitrile and hydra-
zide. These procedures (Scheme 1; path b–d) are usually
conducted at elevated temperature and involve the acti-
vation of nitrile to acylamidrazone intermediate 2 prior
to cyclization. These conventional procedures not only
involve high reaction temperature and long reaction
time but also results in low yields of the product [24–26].
Herein, we are reporting a simple and scaleable method-
ology for the one-pot synthesis of 3,5-disubstituted-1,2,4-
triazoles.
Heterocycles play an important role in all spheres of life
including pharmaceuticals, natural resources, veteri-
nary, agriculture products, analytical reagents, and dyes
[1]. The development of new approaches for the synthesis
of heterocycles decorated with unique functional groups
forms the basis of an extensive research activity in syn-
thetic organic chemistry. Justification of much of the
chemistry directed to the synthesis of the compounds,
possessing nitrogen at the ring fusion, is due to the appli-
cation of compounds having interesting biological prop-
erties in the field of medicinal chemistry. The 1,2,4-tria-
zole moiety is a structural element in certain anti-asth-
matic [2], antiviral (ribavirin) [3], antifungal (flucanoa-
zole) [4], antibacterial [5], hypotonic (triazolam) [6] drugs.
Certain compounds containing 1,2,4-triazole nucleus
have been reported to possess bactericidal [7], antiviral
[8], insecticidal [9], anticancer [10], anti-inflammatory
[11], anticonvulsant [12, 13] properties. Also, some tria-
zole derivatives have been synthesized as plant-growth
regulators [14].
To the best of our knowledge, 3,5-disubstituted-1H-
1,2,4-triazoles have not yet been reported from long-
chain saturated and olefinic carboxylic acids. The present
work is in continuation of our study on the synthesis of
heterocycles from such acids. Tetrazoles, [27, 28] pyrazo-
lines [29], tetrazines [30, 31], spiro [oxathiolane-2,29-dihy-
drotetrazoles] [32], aziridines [33], and triazines [34] have
Correspondence: Abdul Rauf, Department of Chemistry, Aligarh Muslim
University, Aligarh 202002, India.
E-mail: abduloafchem@gmail.com
Fax: +91 5712702885
i 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Arch. Pharm. Chem. Life Sci. 2008, 341, 714–720
3,5-Disubstituted-1H-1,2,4-triazoles
715
Scheme 1. Pathways for synthesis of 3,5-disubstituted-1H-
1,2,4-triazoles.
Scheme 2. Synthesis of 3,5-disubstituted-1H-1,2,4-triazoles
5a–o.
been previously prepared in our lab. Cyanoethoxy and let for two hydrogens was observed at d = 2.91 for the
morpholine derivatives of hydroxy long-chain acids [35] methylene protons, alpha to the triazole ring. The struc-
and fatty esters [36] showed significant antifungal and ture of 5g was further supported by its mass spectral stud-
antibacterial activity. In view of the above mentioned ies, which showed a molecular ion peak at m/z 299, con-
pharmacological applications of 1,2,4-triazoles, we con- sistent with its molecular formula C18H25ON3. The base
sidered undertaking the design and synthesis of hitherto peak appears at m/z 160. Detailed spectral of the titled
unknown 1,2,4-triazoles bearing a long alkyl and alkenyl compounds are given in Experimental.
chain.
Biological studies
All the newly synthesized compounds were evaluated in
vitro against an assortment of two Gram-positive bacteria
Results and discussion
Chemistry
Staphylococcus aureus MSSA 22 and Bacillus subtilis ATCC
The 3,5-disubstituted-1H-1,2,4-triazoles 5a–o were syn- 6051 and two Gram-negative bacteria Escherichia coli K 12
thesized by the condensation of long-chain saturated and and Salmonella typhimurium MTCC 98 at a concentration
olefinic carboxylic acid hyrazides 3a–e with nitriles 4 in of 100 lg/mL. Chloramphenicol was used as standard
presence of a catalytic amount of K2CO3 in n-BuOH drug for the comparison of the antibacterial results.
(Scheme 2). The use of a catalytic amount of K2CO3 pro- Screening results are summarized in Table 2. The newly
vided the product in reduced reaction time in appreci- generated compounds 5a–o have exerted significant
able yield as compared to methods reported by earlier inhibitory activity against the growth of the tested bacte-
workers. As can been seen from Table 1, the scope of the rial strains. The data pertaining to Table 3 reveal that
reaction using saturated, olefinic (internal and terminal), 5a–o have significant influence on the antibacterial pro-
and hydroxy acid hydrazides was found to be good. The file of S. typhimurium and S. aureus. The synthesized com-
yields of 3,5-disubstituted-1H-1,2,4-triazoles did not pounds showed good inhibitory results against B. subtilis
depend on the length of chain of the acid hydrazide and and E. coli. In another set of experiments, the above men-
were found to be appreciable. The synthesized com- tioned compounds 5a–o were also examined for their
pounds were identified on the basis of elemental anal- antifungal activity. Nystatin was used as standard drug
ysis, IR, 1H-NMR, 13C-NMR, and mass spectra. The 1H-NMR for the comparison of the antifungal results. The synthe-
spectrum of 3-(49-hydroxyphenyl)-5-(dec-9-enyl)-1H-1,2,4- sized compounds showed excellent inhibitory results for
triazole 5g showed characteristic signals of d = 10.97 of C. albicans IOA-109 and good results against Penicillium sp.
the -NH proton and multiplets at d = 7.57–7.53 for four (laboratory isolate) and Helminthosporum oryzae (2537 lab-
aromatic protons. The methine proton of C-10 showed oratory isolate). All compounds showed moderate activ-
signals at d = 5.82. C-11 methylene designated as HE ity against Trichoderma. viridae (laboratory isolate) and
and Hz displayed two distinct d values when coupled Aspergillus niger (laboratory isolate) (Table 3). The data
1
with the adjacent C-10 methine protons. Thus, the H also revealed that 5a–o has produced the marked
NMR spectrum showed two doublets of a doublet at d = enhancement in the potency of these analogues as anti-
5.02 and 4.90 for Hz and HE protons, respectively. A trip- fungal and antibacterial agents.
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Arch. Pharm. Chem. Life Sci. 2008, 341, 714–720
Table 1. 3,5-Disubstituted-1H-1,2,4-triazoles 5a–o.
Entry
Starting from
R3
R4
R5
Products
Yield (%)
1
2
3a, 4a
3b, 4a
H
H
H
H
5a
5b
89
85
3
4
3c, 4a
3d, 4a
H
H
H
H
5c
82
82
5d
5
3e, 4a
H
H
5e
80
6
7
8
9
3a, 4b
3b, 4b
3c, 4b
3d, 4b
H
H
H
H
OH
OH
OH
OH
5f
81
88
85
83
5g
5h
5i
10
3e, 4b
H
OH
5j
80
11
12
13
14
3a, 4c
3b, 4c
3c, 4c
3d, 4c
OH
OH
OH
OH
OH
OH
OH
OH
5k
5l
81
80
79
79
5m
5n
15
3e, 4c
OH
OH
5o
78
Conclusion
Experimental
Anhydrous conditions were achieved by drying flasks and other
equipments in the oven. Reactions were monitored by TLC on
silica gel G. Silica gel (60–80 mesh) was used for column chroma-
tography. All reagents and solvents were generally used as
received from commercial suppliers and when required, sol-
vents were dried and distilled before use. Undec-10-enoic, (Z)-
octadec-9-enoic, and octadecanoic acids were obtained commer-
cially from Fluka Chemicals (Switzerland). The eluent was a mix-
ture of petroleum ether / ethyl acetate in different proportions
for different compounds and was visualized in an iodine cham-
ber. (9Z, 12R)-12-Hydroxyoctadec-9-enoic (ricinoleic) and (9R,
12Z)-9-hydroxyoctadec-12-enoic (isoricinoleic) acids were iso-
lated from the natural sources, i.e. from Ricinus communis and
Wrightia tinctoria seed oils, respectively. The concentrate of pure
hydroxy acids were obtained by Gunstone's partitioning [37] of
freshly prepared fatty acids and further purified by column
chromatography. 1H-NMR spectra were recorded in CDCl3 on a
The preparation of 3,5-disubstituted-1H-1,2,4-triazole
derivatives of long-chain alkyl and alkenyl acid hydra-
zides is a valuable addition to the heterocyclic chemistry.
In conclusion, we have developed a useful procedure for
the synthesis of 3,5-disubstituted-1H-1,2,4-triazole from
fatty acid hydazides which could be scaled up to large
quantities. Further biological evaluation of these deriv-
atives may prove potential usefulness.
We are thankful to the SAIF, Panjab University, Chandigarh,
and the Central Drug Research Institute, Lucknow, for provid-
ing spectral data.
The authors have declared no conflict of interest.
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Arch. Pharm. Chem. Life Sci. 2008, 341, 714–720
3,5-Disubstituted-1H-1,2,4-triazoles
717
Table 2. Antibacterial screening data for 3,5-disubstituted-1H-1,2,4-triazoles 5a–o.
Compound
Diameter of zone of inhibition (mm) at 100 lg/mL
Gram-negative
Gram-positive
ECa)
STb)
BSc)
SAd)
5a
5b
5c
5d
5e
5f
5g
5h
5i
5j
5k
5l
5m
5n
5o
14.6 l 0.61
14.2 l 0.64
13.6 l 0.53
13.2 l 0.53
13.2 l 0.50
16.1 l 0.98
15.9 l 0.36
15.4 l 0.61
13.3 l 0.75
13.1 l 0.65
16.6 l 0.59
16.6 l 0.60
15.9 l 0.65
14.0 l 0.91
14.2 l 0.53
25
14.6 l 0.42
19.1 l 0.66
18.2 l 0.51
17.1 l 0.23
16.6 l 0.53
16.5 l 0.50
19.1 l 0.25
18.5 l 0.46
18.1 l 0.31
17.1 l 0.63
16.3 l 0.50
20.3 l 0.42
20.2 l 0.53
19.7 l 0.31
19.4 l 0.40
19.2 l 0.35
24
15.4 l 0.51
15.5 l 0.50
16.7 l 0.31
15.8 l 0.60
16.6 l 0.53
16.9 l 0.83
16.9 l 0.34
17.8 l 0.72
18.5 l 0.42
18.4 l 0.53
20.3 l 0.46
20.9 l 0.90
21.3 l 0.64
22.5 l 0.50
22.2 l 0.81
26
15.3 l 0.42
15.6 l 0.53
16.5 l 0.42
16.3 l 0.46
17.1 l 0.23
16.7 l 0.61
16.2 l 0.29
17.1 l 0.42
17.4 l 0.41
18.2 l 0.53
18.5 l 0.31
17.1 l 0.12
16.7 l 0.61
17.2 l 0.35
20
Chlor amphenicol
Control DMSO
–
–
–
–
a)
EC: Escherichia coli.
ST: Salmonella typhimurium.
BS: Bacillus subtilis.
b)
c)
d)
SA: Staphylococcus aureus.
Table 3. Antifungal screening data for 3,5-disubstituted-1H-1,2,4-triazoles 5a–o.
Compound
Diameter of zone of inhibition (mm) at 100 lg/mL
CAa)
HOb)
ANc)
TVd)
PNe)
5a
5b
5c
5d
5e
5f
5g
5h
5i
5j
5k
5l
5m
5n
5o
18.37 l 0.31
18.13 l 0.32
18.3 l 0.55
17.2 l 0.49
17.2 l 0.35
19.03 l 0.44
18.9 l 0.55
18.03 l 0.68
17.6 l 0.38
18.03 l 0.85
18.9 l 0.40
18.2 l 0.43
17.7 l 0.44
18.4 l 0.40
17.9 l 0.57
20
12.3 l 0.36
12.5 l 0.50
11.2 l 0.49
10.7 l 0.61
10.8 l 0.71
13.1 l 0.57
13.2 l 0.46
14.0 l 0.47
14.2 l 0.47
14.2 l 0.60
13.6 l 0.40
13.9 l 0.75
13.9 l 0.46
14.1 l 0.60
14.7 l 0.55
18
15.0 l 0.25
14.7 l 0.86
15.1 l 0.42
14.4 l 0.40
14.8 l 0.80
15.1 l 0.42
14.8 l 0.91
15.1 l 0.55
15.7 l 0.42
15.5 l 0.50
16.1 l 0.40
16.0 l 0.50
15.7 l 0.80
16.1 l 0.48
16.0 l 0.30
18
5.1 l 0.42
5.4 l 0.46
5.6 l 0.60
5.5 l 0.50
5.4 l 0.58
6.5 l 0.50
6.7 l 0.42
6.7 l 0.61
6.8 l 0.50
6.9 l 0.59
8.1 l 0.50
8.2 l 0.43
8.8 l 0.25
8.7 l 0.50
9.2 l 0.62
15
12.5 l 0.50
12.7 l 0.61
12.9 l 0.80
13.2 l 0.53
13.2 l 0.50
14.6 l 0.55
14.7 l 0.56
15.2 l 0.43
15.2 l 0.58
15.6 l 0.40
16.0 l 0.30
16.5 l 0.55
17.2 l 0.50
17.0 l 0.36
17.4 l 0.43
20
Nystatin
Control DMSO
–
–
–
–
–
a)
CA: Candida albicans.
b)
HO: Helminthosporum oryza.
AN: Aspergillus niger.
TV: Trichoderma viridae.
PN: Penicillium sp.
c)
d)
e)
Bruker DRX-400 instrument Bruker Bioscience, Billerica, MA,
USA). The chemical shifts (d) were measured relative to TMS as
an internal standard. Coupling constants (J) are expressed in Hz.
Mass spectra were obtained on a Jeol SX-102 (FAB) spectrometer
(JEOL, Tokyo, Japan). IR spectra were obtained on Shimadzu
8201 PC FT-IR using KBr pellet with absorption given in cm– 1
(Shimadzu, Tokyo, Japan).
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Arch. Pharm. Chem. Life Sci. 2008, 341, 714–720
Synthesis
3-Pheny-5-[(8R, 11Z)-8-Hydroxyheptadec-11-enyl]-1H-
1,2,4-triazole 5e
General procedure for preparation of 3,5-disubstituted-
1H-1,2,4-triazoles from fatty acid hydrazides
IR (KBr): 3387 (N-H), 1594 (C=N), 1124 (C-N) cm– 1 1H-NMR
;
(400 MHz, CDCl3) d (ppm): 10.98 (s, 1H, -NH-), 7.69–7.65 (m, 2H,
Ar-H), 7.62–7.55 (m, 1H, Ar-H), 7.49–7.45 (m, 2H, Ar-H), 5.36 (m,
2H, -CH=CH-), 3.88 (m, 1H, CHOH), 3.11 (t, 2H, J = 7.6 Hz, -CH2 a to
ring), 2.42 (m, 1H, CHOH), 2.03 (m, 4H, CH2-CH=CH-CH2), 1.98 (m,
2H, -CH2 b to ring), 1.29 (brs, 18H, chain CH2), 0.88 (dist.t, 3H,
CH3). 13C-NMR (100 MHz, CDCl3) d (ppm): 157.2, 146.8, 134.7,
131.5, 129.5, 127.2, 67.3, 39.1, 34.6, 31.8, 29.1, 28.7, 27.1, 22.5,
14.7. MS, m/z (%): [M + 1]+ 398 (5.8), [M]+ 397 (31.2), 368 (33.2), 326
(13.1), 272 (26.1), 228 (19.9), 214 (100), 172 (30.0).
The hydrazides 3 of corresponding long-chain acids were pre-
pared as reported earlier [34]. A mixture of nitrile (3 mmol) 4,
acid hydrazide 3a-e (1 mmol), and K2CO3 (0.5 mmol) in n-BuOH
(2 mL) was stirred and refluxed at 1508C for 4 hours. The prog-
ress of the reaction was monitored on TLC. After completion of
the reaction, the solvent was removed under reduced pressure
and the compounds were adsorbed on silica gel and purified by
column chromatography. All the compounds 5a–o were
obtained as oily liquids.
3-(49-Hydroxyphenyl)-5-heptadecyl-1H-1,2,4-triazole 5f
3-Phenyl-5-heptadecyl-1H-1,2,4-triazole 5a
IR (KBr): 3387 (N-H), 1590 (C=N), 1132 (C-N) cm– 1 1H-NMR
;
IR (KBr): 3382 (N-H), 1607 (C=N), 1123 (C-N) cm– 1 1H-NMR
;
(400 MHz, CDCl3) d (ppm): 10.97 (s, 1H, -NH-), 7.57–7.53 (m, 4H,
Ar-H), 6.95 (Ar-OH), 2.78 (t, 2H, J = 7.6 Hz, -CH2 a to ring), 1.92 (m,
2H, -CH2 b to ring), 1.23 (brs, 28H, chain CH2), 0.88 (dist.t, 3H,
CH3). 13C-NMR (100 MHz, CDCl3) d (ppm): 159.1, 151.8, 148.7,
138.9, 129.7, 117.3, 32.5, 30.7, 22.8, 21.7, 14.5. MS, m/z (%): [M +
1]+ 400 (13.8), [M]+ 399 (41.6), 286 (16.6), 272 (83.3), 258 (16.6) 188
(8.3), 174 (44.4), 160 (100).
(400 MHz, CDCl3) d (ppm): 10.96 (s, 1H, -NH-), 7.69–7.67 (m, 2H,
Ar-H), 7.65–7.60 (m, 1H, Ar-H), 7.49–7.42 (m, 2H, Ar-H), 2.96 (t, J =
7.6 Hz, 2H, -CH2 a to ring), 2.05 (m, 2H, -CH2 b to ring), 1.72 (brs,
28H, chain CH2), 0.86 (dist. t, 3H, CH3). 13C-NMR (100 MHz, CDCl3)
d (ppm): 159.1, 147.9, 133.8, 129.1, 127.3, 32.1, 30.3, 29.9, 28.6,
23.1, 14.8. MS, m/z (%): [M + 1]+ 384 (10.5), [M]+ 383 (36.11), 354
(25.3), 270 (27.7), 214 (63.8), 186 (16.6), 58 (100).
3-(49-Hydroxyphenyl)-5-(dec-9-enyl)-1H-1,2,4-triazole 5g
IR (KBr): 3390 (N-H), 1590 (C=N), 1129 (C-N) cm– 1 1H-NMR
(400 MHz, CDCl3) d (ppm): 10.97 (s, 1H, -NH-), 7.57–7.53 (m, 4H,
3-Phenyl-5-(dec-9-enyl)-1H-1,2,4-triazole 5b
IR (KBr): 3392 (N-H), 1594 (C=N), 1122 (C-N) cm– 1 1H-NMR
;
(400 MHz, CDCl3) d (ppm): 10.98 (s, 1H, -NH-), 7.66–7.64 (m, 2H,
Ar-H), 7.62–7.58 (m, 1H, Ar-H), 7.49–7.45 (m, 2H, Ar-H), 5.82 (tdd,
8
Ar-H), 6.93 (Ar-OH), 5.82 (tdd, 1H, JHÀ CH = 6.6 Hz, JHÀH = 10.2 Hz,
Z
2
JHÀH = 17.1 Hz, CH2=CH-), 5.02 (dd, 1H, JH ÀH= 10.2 Hz, JH ÀH
=
E
E
E
Z
8
1H, JHÀ CH = 6.6 Hz, JHÀH = 10.2 Hz, JHÀH = 17.1 Hz, CH2=CH-), 5.02
Z
E
2
1.2 Hz, HZ C=CH-), 4.90 (dd, 1H, J
= 17.1 Hz, JH ÀH = 1.2 Hz, HE
HEÀH
E
Z
(dd, 1H, JH ÀH= 10.2 Hz, JH ÀH = 1.2 Hz, HZ C=CH-), 4.90 (dd, 1H,
JHÀH = 17.1 Hz, JH ÀH = 1.2 Hz, HE C=CH-), 3.10 (t, 2H, J = 7.6 Hz,
Z
z
E
C=CH-), 2.91 (t, 2H, J = 7.9 Hz, -CH2 a to ring), 2.02 (m, 2H, -CH2-
CH=CH2), 1.93 (m, 2H, -CH2 b to ring), 1.38 (brs, 10H, chain CH2).
E
E
Z
-CH2 a to ring), 2.05 (m, 2H, CH2-CH=CH2), 1.99 (m, 2H, -CH2 b to
ring), 1.38 (10H, brs, chain CH2). 13C-NMR (100 MHz, CDCl3) d
(ppm): 158.9, 147.2, 139.8, 133.9, 129.6, 127.6, 114.6, 32.8, 30.8,
29.2, 28.3, 22.9, 14.2. MS, m/z (%): [M + 1]+ 284 (14.7), [M]+ 283
(10.8), 270 (28.3), 242 (29.9), 228 (11.8), 214 (18.3), 186 (26.2), 144
(100).
13C-NMR (100 MHz, CDCl3) JH ÀH (ppm): 158.7, 151.2, 148.1, 139.7,
E
129.1, 116.2, 114.4, 34.2, 33.7, 29.2, 24.7. MS, m/z (%): [M + 1]+ 300
(19.9), [M]+ 299 (12.3), 272 (12.5), 244 (17.3), 230 (3.9), 216 (2.2),
188 (3.9), 174 (4.2), 160 (100).
3-(49-Hydroxyphenyl)-5-(heptadec-8-enyl)-1H-1,2,4-
triazole 5h
3-Phenyl-5-(heptadec-8-enyl)-1H-1,2,4-triazole 5c
IR (KBr): 3387(N-H), 1594 (C=N), 1130 (C-N) cm– 1 1H-NMR
;
IR (KBr): 3382 (N-H), 1597 (C=N), 1133 (C-N) cm– 1 1H-NMR
;
(400 MHz, CDCl3) d (ppm): 10.97 (s, 1H, -NH-), 7.57–7.56 (m, 4H,
Ar-H), 6.93 (Ar-OH), 5.39 (2H, m, -CH=CH-), 2.79 (t, 2H, J = 5.7 Hz,
-CH2 a to ring), 2.02 (m, 4H, -CH2-CH=CH-CH2-), 1.96 (m, 2H, -CH2 b
to ring), 1.23 (brs, 20H, chain CH2), 0.88 (dist.t, 3H, CH3). 13C-NMR
(100 MHz, CDCl3) d (ppm): 157.7, 151.2, 148.7, 130.8, 129.1,
118.4, 34.2, 31.8, 29.6, 29.1, 27.0, 24.7, 22.5, 14.6. MS, m/z (%): [M
+ 1]+ 398 (13.8), [M]+ 397 (41.6), 284 (16.6), 258 (16.6) 188 (8.3), 174
(100), 160 (44.4).
(400 MHz, CDCl3) d (ppm): 10.96 (s,1H, -NH-), 7.67–7.63 (m, 2H,
Ar-H), 7.60-7.56 (m, 1H, Ar-H), 7.49–7.44 (m, 2H, Ar-H), 5.34 (2H,
m, -CH=CH-), 2.97 (t, 2H, J = 7.6 Hz, -CH2 a to ring), 2.05 (m, 4H,
CH2-CH=CH-CH2), 1.96 (m, 2H, -CH2 b to ring), 1.73 (brs, 20H,
chain CH2), 0.86 (dist.t, 3H, CH3). 13C-NMR (100 MHz, CDCl3) d
(ppm): 158.2, 146.9, 134.1, 131.1, 129.0, 128.1, 33.8, 30.8, 29.9,
28.6, 22.6, 14.6. MS, m/z (%): [M + 1]+ 382 (40.5), [M]+ 381 (30.11),
338 (38.3), 310 (27.7), 268 (63.8), 242 (16.6), 158 (100), 144 (10.9).
3-Phenyl-5-[(8Z, 11R)-11-hydroxyheptadec-8-enyl]-1H-
3-(49-Hydroxyphenyl)-5-[(8Z, 11R)-11-hydroxyheptadec-
8-enyl]-1H-1,2,4-triazole 5i
1,2,4-triazole 5d
IR (KBr): 3386 (N-H), 1590 (C=N), 1119 (C-N) cm– 1
;
1H-NMR
IR (KBr): 3390 (N-H), 1590 (C=N), 1123 (C-N) cm– 1 1H-NMR
;
(400 MHz, CDCl3) d (ppm): 10.99 (s, 1H, -NH-), 7.66–7.64 (m, 2H,
Ar-H), 7.60–7.57 (m, 1H, Ar-H), 7.49–7.46 (m, 2H, Ar-H), 5.34 (m,
2H, -CH=CH-), 3.88 (m, 1H, CHOH), 3.11 (t, 2H, J = 7.6 Hz, -CH2 a to
ring), 2.43 (m, 1H, CHOH), 2.05 (m, 4H, CH2-CH=CH-CH2), 1.89 (m,
2H, -CH2 b to ring), 1.73 (brs, 18H, chain CH2), 0.89 (dist.t, 3H,
CH3). 13C-NMR (100 MHz, CDCl3) d (ppm): 157.9, 146.8, 134.0,
131.6, 129.2, 127.9, 67.8, 39.2, 34.6, 31.2, 29.4, 28.7, 27.1, 22.7,
14.9. MS, m/z (%): [M + 1]+ 398 (9.8), [M]+ 397 (21.2), 340 (13.2), 312
(13.1), 282 (26.8), 200 (39.9), 186 (100), 172 (30.0).
(400 MHz, CDCl3) ( (ppm): 10.98 (s, 1H, -NH-), 7.58–7.52 (m, 4H,
Ar-H), 6.98 (Ar-OH), 5.46 (m, 2H, -CH=CH-), 3.88 (m, 1H, -CH-OH),
3.44 (t, 2H, J = 7.5 Hz, -CH2 a to ring), 2.31 (m, 1H, -CH-OH), 2.04
(m, 4H, -CH2-CH=CH-CH2-), 1.91 (m, 2H, -CH2 b to ring), 1.33 (brs,
18H, chain CH2), 0.86 (dist.t, 3H, CH3). 13C-NMR (100 MHz, CDCl3)
d (ppm): 157.9, 151.6, 148.5, 130.8, 129.3, 118.7, 67.8, 39.2, 34.1,
31.8, 29.6, 28.9, 27.2, 24.7, 22.5, 14.3. MS, m/z (%): [M + 1]+ 414
(12.2), [M]+ 413 (26.13), 370 (75.8), 356 (60.9), 328 (33.4), 202 (69.8),
174 (14.2), 160 (100).
i 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

Arch. Pharm. Chem. Life Sci. 2008, 341, 714–720
3,5-Disubstituted-1H-1,2,4-triazoles
719
-CH-OH), 3.44 (t, 2H, J = 7.5 Hz, -CH2 a to ring), 2.31 (m, 1H, -CH-
OH), 2.04 (m, 4H, -CH2-CH=CH-CH2-), 1.91 (m, 2H, -CH2 b to ring),
1.33 (brs, 18H, chain CH2), 0.86 (dist.t, 3H, CH3). 13C-NMR
(100 MHz, CDCl3) (ppm): 163.9, 147.8, 145.1, 131.8, 121.1, 119.2,
68.2, 39.8, 37.2, 33.1, 29.9, 27.6, 24.1, 22.7, 14.3. MS, m/z (%): [M +
1]+ 430 (6.6), [M]+ 429 (28.3), 344 (43.3), 260 (13.8), 253 (41.6), 208
(8.3), 190 (47.2), 176 (100).
3-(49-Hydroxyphenyl)-5-[(8R, 11Z)-8-Hydroxyheptadec-
11-enyl]-1H-1,2,4-triazole 5j
IR (KBr): 3379 (N-H), 1592 (C=N), 1119 (C-N) cm– 1 1H-NMR
;
(400 MHz, CDCl3) d (ppm): 10.99 (s, 1H, -NH-), 7.57-7.53 (m, 4H, Ar-
H), 6.93 (Ar-OH), 5.39 (m, 2H, -CH=CH-), 3.88 (m, 1H, -CH-OH), 3.23
(t, 2H, J = 6.4 Hz, -CH2 a to ring), 2.28 (m, 1H, -CH-OH), 2.04 (m, 4H,
-CH2-CH=CH-CH2-), 1.89 (m, 2H, -CH2 b to ring), 1.27 (brs, 18H,
chain CH2), 0.86 (dist.t, 3H, CH3). 13C-NMR (100 MHz, CDCl3) d
(ppm): 158.9, 151.6, 148.7, 131.1, 129.4, 118.6, 67.3, 39.6, 34.1,
31.8, 29.5, 28.7, 27.2, 24.5, 22.1, 14.4. MS, m/z (%): [M + 1]+ 414
(12.5), [M]+ 413 (29.1), 342 (51.2), 316 (33.3), 288 (9.8), 188 (100),
174 (18.8), 160 (80.2).
3-(49,59-Dihydroxyphenyl)-5-[(8R, 11Z)-8-
hydroxyheptadec-11-enyl]-1H-1,2,4-triazole 5o
IR (KBr): 3392 (N-H), 1600 (C=N), 1123 (C-N) cm– 1 1H-NMR
;
(400 MHz, CDCl3) d (ppm): 10.98 (s, 1H, -NH-), 7.11 (d, 1H, J =
1.6 Hz, Ar-H), 7.04 (d, 1H, J = 2 Hz, Ar-H), 7.02 (d, 1H, J = 2 Hz, Ar-
H), 6.89 (Ar-OH), 6.87 (Ar-OH), 5.39 (m, 2H, -CH=CH-), 3.88 (m, 1H,
-CH-OH), 3.23 (t, 2H, J = 6.4 Hz, -CH2 a to ring), 2.28 (m, 1H, J =
7.2 Hz, -CH-OH), 2.04 (m, 4H, -CH2-CH=CH-CH2-), 1.89 (m, 2H, -CH2
b to ring), 1.27 (brs, 18H, chain CH2), 0.86 (dist.t, 3H, CH3). 13C-
NMR (100 MHz, CDCl3) d (ppm): 163.6, 147.5, 145.1, 131.7, 121.2,
119.6, 68.4, 39.8, 37.4, 33.1, 29.5, 27.3, 24.2, 22.4, 14.2. MS, m/z
(%): [M + 1]+ 430 (55.5), 372 (55.5), [M]+ 429 (8.3), 332 (41.6), 301
(22.2), 280 (13.6), 208 (8.3), 176 (100).
3-(49,59-Dihydroxyphenyl)-5-(heptadecyl)-1H-1,2,4-
triazole 5k
IR (KBr): 3349 (N-H), 1604 (C=N), 1134 (C-N) cm– 1 1H-NMR
;
(400 MHz, CDCl3) d (ppm): 10.97 (s, 1H, -NH-), 7.16 (d, 1H, J =
1.6 Hz, Ar-H), 7.07 (d, 1H, J = 2 Hz, Ar-H), 7.03 (d, 1H, J = 2 Hz, Ar-
H), 6.89 (Ar-OH), 6.87 (Ar-OH), 2.78 (t, 2H, J = 7.6 Hz, -CH2 a to
ring), 1.92 (m, 2H, -CH2 b to ring), 1.23 (brs, 28H, chain CH2), 0.88
(dist.t, 3H, CH3). 13C-NMR (100 MHz, CDCl3): 164.5, 145.1, 144.2,
121.9, 115.9, 32.1, 30.3, 28.9, 22.3, 14.5. MS, m/z (%): [M + 1]+ 416
(17.0), [M]+ 415 (66.0), 356 (10.4), 314 (2.1), 300 (31.2), 208 (19.2),
190 (12.5), 176 (100).
Biological evaluation
Antibacterial activity
The newly synthesized compounds were screened in vitro against
an assortment of two Gram-positive bacteria Staphylococcus aur-
eus MSSA 22 and Bacillus subtilis ATCC 6051 and two Gram-nega-
tive bacteria Escherichia coli K12 and Salmonella typhimurium
MTCC 98. Screening results are summarized in Table 2. All the
synthesized compounds were dissolved in DMSO. The antibacte-
rial activity of test compounds and standard chloramphenicol
was done by filter paper disc method [38]. Media with DMSO was
set up as control. All cultures were routinely maintained on NA
(nutrient agar) and incubated at 378C. The inoculums of bacteria
were performed by growing the culture in NA broth at 378C over-
night. The culture was centrifuged at 1000 rpm, pellets were
resuspended, and diluted in sterile NSS to obtain viable count
105 CFU/mL. With the help of spreader, 0.1 mL of approximately
diluted bacterial culture suspension was spread on NA plates
uniformly. Sterile 8-mm discs (Hi-media Pvt. Ltd.) were impreg-
nated with 100 lg/mL concentration of the test compounds.
Antibiotic disc, chloramphenicol (30 lg/disc, Hi-Media) was used
as control. The disc was placed onto the plate. Each plate had
one control disc impregnated with solvent. The plates were then
incubated for 24 h at 378C, and the resulting zones of inhibition
(in mm) were measured. Diameters of the zone of inhibition
(mm) were measured and the average diameters for the test sam-
ples were calculated in triplicate sets.
3-(49,59-Dihydroxyphenyl)-5-(dec-9-enyl)-1H-1,2,4-
triazole 5l
IR (KBr): 3387 (N-H), 1596 (C=N), 1130 (C-N) cm– 1 1H-NMR
;
(400 MHz, CDCl3) d (ppm): 10.98 (s, 1H, -NH-), 7.11 (d, 1H, J =
1.6 Hz, Ar-H), 7.04 (d, 1H, J = 2 Hz, Ar-H), 7.02 (d, 1H, J = 2 Hz, Ar-
8
H), 6.88 (Ar-OH), 6.87 (Ar-OH), 5.82 (tdd, 1H, JHÀ CH = 6.6 Hz,
2
JHÀH = 10.2 Hz, JHÀH = 17.1 Hz, CH2=CH-), 5.02 (dd, 1H, J
=
HZÀH
Z
E
10.2 Hz, JH ÀH = 1.2 Hz, HZ C=CH-), 4.90 (dd, 1H, J
= 17.1 Hz,
HEÀH
Z
E
JH ÀH = 1.2 Hz, HE C=CH), 3.13 (t, 2H, J = 8 Hz, -CH2 a to ring), 2.02
E
Z
(m, 2H, -CH2-CH=CH2), 1.93 (m, 2H, -CH2 b to ring), 1.38 (brs, 10H,
chain CH2). 13C-NMR (100 MHz, CDCl3) d (ppm): 164.9, 145.8,
144.8, 139.9, 121.9, 116.7, 114.6, 34.2, 33.7, 29.2, 28.4, 24.2. MS,
m/z (%): [M + 1]+ 316 (25.7), [M]+ 315 (23.8), 288 (44.2), 274 (66.6),
243 (17.1), 208 (36.3), 198 (14.6), 176 (100).
3-(49,59-Dihydroxyphenyl)-5-(heptadec-8-enyl)-1H-1,2,4-
triazole 5m
IR (KBr): 3384 (N-H), 1594 (C=N), 1127 (C-N) cm– 1 1H-NMR
;
(400 MHz, CDCl3) d (ppm): 10.97 (s, 1H, -NH-), 7.11 (d, 1H, J =
1.6 Hz, Ar-H), 7.04 (d, 1H, J = 2 Hz, Ar-H), 7.02 (d, 1H, J = 2 Hz, Ar-
H), 6.93 (Ar-OH), 6.89 (Ar-OH), 5.34 (m, 2H, -CH=CH-), 2.78 (t, 2H, J
= 7.6 Hz, -CH2 a to ring), 2.02 (m, 4H, -CH2-CH=CH-CH2-), 1.92 (m,
2H, -CH2 b to ring), 1.23 (brs, 20H, chain CH2), 0.88 (dist.t, 3H,
CH3). 13C-NMR (100 MHz, CDCl3) d (ppm): 163.4, 146.8, 144.8,
131.9, 121.6, 119.8, 33.1, 29.9, 25.6, 22.7, 14.9. MS, m/z (%): [M +
1]+ 414 (17.0), [M]+ 413 (66.0), 356 (10.4), 314 (12.1), 300 (31.2), 208
(19.2), 190 (100), 176 (57.5).
Antifungal activity
The standard agar disc diffusion method [38] was performed to
evaluate the antifungal property of the test compounds and
standard nystatin. The newly synthesized compounds were
screened for Aspergillus niger (laboratory isolate), Candida albi-
cans IOA-109, Penicillium sp. (laboratory isolate), Trichoderma viri-
dae (laboratory isolate), Helminthosporum oryzae (2537 ICAR, Jai-
pur); see Table 3. The synthesized compounds were dissolved in
DMSO. Media with DMSO was set up as control. All cultures were
routinely maintained on SDA and incubated at 288C. Spore for-
mation of filamentous fungi was prepared from seven-day old
culture in sterile normal solution (8% NaCl) and diluted to
3-(49,59-Dihydroxyphenyl)-5-[(8Z, 11R)-11-
hydroxyheptadec-8-enyl]-1H-1,2,4-triazole 5n
IR (KBr): 3386 (N-H), 1597 (C=N), 1123 (C-N) cm– 1 1H-NMR
;
(400 MHz, CDCl3) d (ppm): 10.98 (s, 1H, -NH-), 7.13 (d, 1H, J =
1.6 Hz, Ar-H), 7.04 (d, 1H, J = 2 Hz, Ar-H), 7.01 (d, 1H, J = 2 Hz, Ar-
H), 6.89 (Ar-OH), 6.86 (Ar-OH), 5.46 (m, 2H, -CH=CH-), 3.88 (m, 1H,
i 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com

720
S. Sharma et al.
Arch. Pharm. Chem. Life Sci. 2008, 341, 714–720
obtain approximately 105 CFU/mL. The inoculum of non-sporing
fungi C. albicans was performed by growing the culture in SD
broth at 378C overnight. The culture was centrifuged at
1000 rpm, pellets was resuspended, and diluted in sterile NSS to
obtain a viable count 105 CFU/ml. With the help of spreader,
0.1 mL of the approximately diluted fungal culture suspension
was spread on SDA plates uniformly. Sterile 8-mm discs (Hi-
media Pvt. Ltd., Mumbai, India) were impregnated with test com-
pounds. Antibiotic disc, nystatin (30 lg/disc Hi-Media) was used
as control. The disc was placed onto the plate. Each plate had
one control disc impregnated with solvent. The plates were incu-
bated at 288C for filamentous fungi for 72 h or more, while for C.
albicans plates were incubated at 378C for 18–48 h. Antifungal
activity was determined by measuring the diameters of the
inhibition zone (mm) in triplicate sets.
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i 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
www.archpharm.com