Anthelmintic evaluation of some novel synthesized 1,2,4-triazole moiety clubbed with benzimidazole ring

Article abstract of DOI:10.13005/ojc/300125

A series of N-[3-{(1H-benzo[d]imidazol-2-ylthio) methyl}-5-mercapto-4H-1,2, 4-triazol-4-yl]-2-substituted phenoxy acetamide 6(a-g) were synthesized by the mixture of the compound of potassium 2-(2-(1H-benzo[d]imidazol-2-ylthio) acetyl) hydrazinecarbodithioate (4) and arytoxy acid hydrazide (5) in presence of hydrochloric acid. The predicted structures of the synthesized compounds were confirmed by different spectral analysis studies. The title compounds 6(a-g) were screened for anthelmintic activity against Pheretima posthumous. The entire compounds were exhibited good anthelmintic activity when compared with standard drugs such as Albendazole and Piperazine.

Full text of DOI:10.13005/ojc/300125

ISSN: 0970-020 X
CODEN: OJCHEG
2014, Vol. 30, No. (1):
Pg. 211-217
ORIENTAL JOURNAL OF CHEMISTRY
An International Open Free Access, Peer Reviewed Research Journal
www.orientjchem.org
Anthelmintic Evaluation of Some Novel Synthesized
1,2,4-Triazole Moiety Clubbed with Benzimidazole Ring
P. SUDHIR KUMAR1* and J. SAHOO2
1School of Pharmaceutical Sciences, Department of Pharmaceutical Chemistry,
Siksha ‘O’ Anusandhan University, Bhubaneswar, Odisha - 751 003, India.
2Research Scholar, School of Pharmaceutical Sciences, Siksha ‘O’ Anusandhan University,
Bhubaneswar, Odisha - 751 003, India.
*Corresponding author E-mail: sairampaidesetty@gmail.com
http://dx.doi.org/10.13005/ojc/300125
(Received: January 25, 2014; Accepted: March 03, 2014)
ABSTRACT
A series of N-[3-{(1H-benzo[d]imidazol-2-ylthio) methyl}-5-mercapto-4H-1, 2, 4-triazol-4-
yl]-2-substituted phenoxy acetamide 6(a-g) were synthesized by the mixture of the compound of
potassium 2-(2-(1H-benzo[d]imidazol-2-ylthio) acetyl) hydrazinecarbodithioate (4) and aryloxy acid
hydrazide (5) in presence of hydrochloric acid.The predicted structures of the synthesized compounds
were confirmed by different spectral analysis studies. The title compounds 6(a-g) were screened
for anthelmintic activity against Pheretima posthumous.The entire compounds were exhibited good
anthelmintic activity when compared with standard drugs such as Albendazole and Piperazine.
Key words: 5-mercapto-1, 2, 4-triazoles, Anthelmintics activity,
Aryloxy substituted phenoxy acetic acid, 2-merkapto benzimidazole.
INTRODUCTION
the compounds bearing heterocyclic nuclei have
received much more attention due to their medicinal
value in the development of novel anthelmintics
drugs. Benzimidazole derivatives have been a wide
range of biological activities, such as analgesic3,
antiinflammatory4, antiparasitic5, antitubercular6,
antiviral7, anticonvulsant8 and antimicrobial activities9-
10. Literature survey indicated that the compounds
consists of 1, 2, 4-triazole ring have been reported
to possess different biological activities such as
antimicrobial11, antifungal12, anti-inflammatory13,
antioxidant14-15, antiviral16, anticancer17 and
There are two important types of worm
infections, those in which the worms live in the
alimentary canal of the host and worms live in other
tissues of the host body. Common examples of
worms1 in the alimentary canal are the tapeworm,
intestinal round worm, trematodes or flukes, tissue
round worm and diseases caused by helminthes are
ascariasis, trichirians, hook worm and tape worm
infection, thread worm infection, schistosomiasis
and giardial infection2. During the last decades

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KUMAR & SAHOO, Orient. J. Chem., Vol. 30(1), 211-217 (2014)
anticonvulsant activity18 depending on the substituent then it was poured into the ice cold water. The solid
in the ring system. In view of such biological reports, obtained was filtered.Yield 63%, m.p .208°C.
the present work is aimed to design and develop the Preparation of 2-(benzimidazolylthio) acetic acid
synthesis of some new potential substituted aryloxy hydrazide (3)
triazolyl Benz[b] imidazole moiety and explores the
evaluation of their in-vitro anthelmintic activity.
To above ester dissolved in appropriate
quantity of ethanol (25ml) hydrazine hydrate
(0.1mole, 5ml)was added and refluxed for 15hrs.
The mixture was kept for 10-12 hours at room
temperature, solid obtained was filtered.Yield 62%,
EXPERIMENTAL
The chemicals and solvents used for the m. p. 236°C.
experimental work were procured from E. Merck,
India, and Qualigens, India. Silica gel G used for Preparation of potassium 2-[2-(1H-benzo[d]
analytical chromatography (TLC) was obtained from imidazol-2-ylthio)acetyl]hydrazinecarbodithioate
E.Merck.Melting points were determined in an open (4)
glass capillary using a Kjeldahl flask containing
A mixture of 2-(benzimidazolylthio) acetic
paraffin and are uncorrected. The proton magnetic acid hydrazide (0.01mole) and (0.01mole,1.60g) of
resonance spectra (1H NMR) were recorded on a potassium salt was taken in a round bottom flask and
Bruker 300 MHz instrument (Bruker, Germany) in 50ml ethanol was added and refluxed for 8-10 hours
dimethylsulfoxide-d6/CDCl3 using tetramethylsilane as and further diluted with ether (30ml).The separated
internal standard. Chemical shifts (δ) are expressed solid was filtered, washed with ether, dried and used
in ppm. The infrared spectra of compounds were directly for the next step without further purification.
recorded in KBr on infrared spectrophotometer
Jasco, Japan.Purity of the compounds was checked Synthesis of Substituted Aryloxy hydrazide
by TLC using silica gel G.
5(a-g)
The substituted acid (0.1mole) and ethanol
(50 ml) were taken with a few drops of concentrated
Preparation of 2-mercapto Benzimidazole (1)
A mixture of o-phenylenediamine 32.4gm sulfuric acid and it was refluxed for 6 hours. The
(0.3mole) potassium ethyl xanthate 52.8gm reaction mixture was concentrated by distilling
(0.33mole), 95% ethanol 300ml and 45ml of water of the excess of ethanol under reduced pressure
in a 1-1, flask was heated under reflux for 3 hours. and treated with a saturated solution of sodium
Noritz (12gm) was then added cautiously and the bicarbonate. The ester obtained used for the
mixture was heated at the reflux temperature for 10 preparation of hydrazide directly. The ester (0.1mole)
minutes. Then Noritz was removed by filtration, the was dissolved in appropriate quantity of ethanol and
filtrate was heated to 60-70 °C, 300ml of warm tap to this hydrazine hydrate (0.1mole) was added. The
water (60- 70 °C) was added followed by 25ml of reaction mixture was taken in a round bottomed flask
acetic acid in 50ml of water with stirring.The product and refluxed for a period of 12-18 hours. Excess of
separated out as glistening white crystal and the ethanol was distilled off under reduced pressure.
mixture was placed in a refrigerator for 3 hours to It was then poured into ice cold water and the
complete the crystallization.The product is collected solid obtained was filtered. It was crystallized from
on a Buchner funnel dried overnight at 40°C. Yield ethanol.
86%, m.p 303-304°C.
Preparation of 3, 4-disubstituted-5-mercapto-1,
Preparation of Ethyl 2(Benzimidazolylthio) 2, 4-triazoles 6(a-g)
acetate (2)
A mixture of the compound potassium
A mixture of 2-mercaptobenzimidazole 2-(2-(1H-benzo[d]imidazol-2-ylthio) acetyl)
(0.1mole), ethylchloroacetate (0.1mole) and hydrazinecarbodithioate (4) and aryloxy acid
potassium carbonate (0.1mole) and acetone hydrazide 5(a-g) were heated in equimolar proportion
(distilled) was taken in a round bottom flask and in an oil bath at 160-1800C.for 5-6 hours when profuse
refluxed for 6-7 hrs. After the completion of reaction, evolution of hydrogen sulphide was observed. The
the mixture was cooled at room temperature and reaction mixture was cooled and diluted with ice

KUMAR & SAHOO, Orient. J. Chem., Vol. 30(1), 211-217 (2014)
213
cold water. On acidification with hydrochloric acid,
the compound was obtained. It was filtered, washed
with water and crystallized from aqueous ethanol
(80%).
0.2, 0.5 and 1mg/mL of each compound in form
suspension. Normal saline served as a control.The
time taken for complete paralysis and death was
noted.The mean paralysis time and mean lethal time
were calculated for each compound (each reading
was taken in triplicate). Paralysis was considered
when no movement was observed even after shaken
vigorously. Death was considered when the worm
loss their motility followed by change in body color.
Statistically analysis was performed using ANOVA
to find the significance of the test.
Anthelmintic activity
The newly synthesized compounds were
tested for anthelmintic activity19 against Pheretima
posthumous (earth worms collected from Ekahabaor
Botanical Garden, Near to Regional plant Resource
Centre, Bhubaneswar, Odisha, India) of nearly equal
size (6 1 cm) were selected randomly for the
experimental study. The worms were acclimatized
to laboratory conditions before experimentation.The
earthworms were divided into four groups of six earth
worms each. Albendazole and Piperazine citrate
diluted with normal saline solution to obtain 0.1, 0.2,
0.5 and 1 mg/mL act as standard and were poured
into Petri dishes.The synthesized compounds were
dissolved in water and add minimal quantity ofTween
80 and make to prepare four concentrations of 0.1,
6a.N-[3-{(1H-benzo[d]imidazol-2-ylthio)methyl}-
5-mercapto-4H-1,2,4-triazol-4-yl]-2-phenoxy
acetamide
IR (KBr) cm-1: 3315 (NH), 3050-2910 (C-H
str., aromatic and aliphatic), 1150 C=S, 1660 (CO of
CONH), 1605 (C=N), 1500 (C-N), 845 (Substituted
1
benzene), H- NMR (CDCl3) δ: 6.95-7.59 (m, 9H,
Ar-H), 10. 4(s, 1H,N-H),8.22 (1H,CONH), 4.62
(d,2H,S-CH2).

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KUMAR & SAHOO, Orient. J. Chem., Vol. 30(1), 211-217 (2014)
6b. N-[3-{(1H-benzo[d]imidazol-2-ylthio) 6e. N-[3-{(1H-benzo[d]imidazol-2-ylthio)
methyl)-5-mercapto-4H-1, 2, 4-triazol-4-yl}-2-(2- methyl)-5-mercapto-4H-1, 2 ,4-triazol-4-yl}-2-(4-
chlorophenoxy)] acetamide
IR (KBr) cm-1:3310(NH), 2950-2910 (C-H
nitrophenoxy)]acetamide
IR (KBr) cm-1:3340(NH), 3050-2910 (C-H
str., aromatic and aliphatic), 2550 (SH) or 1175 str., aromatic and aliphatic), 2535 (SH) or 1173
C=S, 1664 (CO of CONH), 1615 (C=N), 1500 (C-N), C=S, 1662 (CO of CONH), 1615 (C=N), 1500 (C-N),
1
650(C-Cl), 835(o-disubstituted benzene), H- NMR 1515, 1355 (Ar-NO2), 840(p-disubstituted benzene),
(CDCl3) δ: 6.95-7.59 (m, 8H, Ar-H), 10. 35 (s, 1H, 1HNMR (CDCl3) δ:7.15-8.16 (m, 8H, Ar-H), 10.45 (s,
N-H), 8. 12(1H, CONH), 4.65 (d, 2H, S-CH2).
H, N-H), 8. 12 (1H, CONH), 4. 42 (d, 2H, S-CH2).
6c. N-[3-{(1H-benzo[d]imidazol-2-ylthio) 6f.N-(3-((1H-benzo[d]imidazol-2-ylthio)methyl)-
methyl)-5-mercapto-4H-1, 2, 4-triazol-4-yl}-2-(2, 5-mercapto-4H-1,2,4-triazol-4-yl)-2-(o-tolyloxy)
4-dichlorophenoxy)] acetamide
IR (KBr) cm-1:3310(NH), 3030-2910 (C-H
acetamide
IR (KBr) cm-1:3335(NH), 3050-2910 (C-H
str., aromatic and aliphatic), 2550 (SH) or 1165 str., aromatic and aliphatic), 2535 (SH) or 1165 C=S,
C=S, 1664 (CO of CONH), 1615 (C=N), 1520 (C-N), 1662 (CO of CONH), 1615 (C=N), 1500 (C-N), 840
1
680(C- Cl), 855(trisubstituted benzene), HNMR (o-disubstituted benzene), 1HNMR (CDCl3) δ: 6.88-
(CDCl3) δ: 7.15-7.59 (m, 7H, Ar-H), 10.43 (s, 1H, 7.56 (m, 8H, Ar-H), 10. 35 (s,1H, N-H), 8. 12 (1H,
N-H), 8. 12(1H, CONH), 4.64 (d, 2H, S-CH2).
CONH), 4.4 (d, 2H, SCH2), 2.52 (s, 3H, CH3).
6d N-[3-{(1H-benzo[d]imidazol-2-ylthio) 6g.N-(3-((1H-benzo[d]imidazol-2-ylthio) methyl)-
methyl)-5-mercapto-4H-1, 2, 4-triazol-4-yl}-2-(4- 5-mercapto-4H-1, 2, 4-triazol-4-yl)-2-(p-tolyloxy)
chlorophenoxy)] acetamide
IR (KBr) cm-1:3330(NH), 3050-2910 (C-H
acetamide
IR (KBr) cm-1: 3335(NH), 3050-2910 (C-H
str., aromatic and aliphatic), 2535 (SH) or 1173 str., aromatic and aliphatic), 1165 C=S, 1662 (CO of
C=S, 1662 (CO of CONH), 1615 (C=N), 1500 (C-N), CONH), 1615 (C=N), 1500 (C-N), 845(p-disubstituted
1
650(C- Cl), 835(p-disubstituted benzene), HNMR benzene), 1HNMR (CDCl3) ´:6.88-7.56 (m, 8H, Ar-H),
(CDCl3) δ: 6.95-7.59 (m, 8H, Ar-H), 10.35 (1H, N-H), 10. 55 (s, 1H, N-H),8. 12 (1H, CONH), 4.64 (2H,
8. 12 (1H, CONH), 4.65 (2H, S-CH2).
S-CH2), 2.52 (s,3H,CH3).
Table 1: Characteristic data of Synthesized compounds
Comp. m.p.
°C
YIELD
(%)
M.F
M. Wt
Elemental Analysis (%)
C
H
N
6a
6b
6c
6d
6e
6f
172
278
276
269
278
273
265
88
72
70
69
80
72
70
C18H16O2N6S2
C18H15O2N6S2Cl
C18H14O2N6S2Cl2
C18H15O2N6S2Cl
C18H15N7O4S2
C19H18O2N6S2
C19H18O2N6S2
412
446
480
446
457
426
426
52.14
(52.09)
48.43
(48.25)
44.91
(44.52)
48.43
(48.25)
47.26
(47.12)
53.52
(53.35)
53.52
3.89
(3.75)
3.36
(3.25)
2.93
(2.75)
3.36
(3.22)
3.22
(3.15)
4.22
(4.17)
4.22
20.37
(20.15)
18.83
(18.74)
17.47
(17.25)
18.83
(18.45)
21.43
(21.25)
19.71
(19.63)
19.71
6g
(53.32)
(4.15)
(19.65)

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KUMAR & SAHOO, Orient. J. Chem., Vol. 30(1), 211-217 (2014)
RESULTS AND DISCUSSION
elemental analyses of the compound is in conformity
with the assigned structure.In similar manner seven
3, 4-diarylsubstituted-5-mercapto-1, 2, 4-triazoles
(5) have been characterized in (Table-1). The
anthelmintics evaluation of compounds (6a-6g) in
earth worms at four concentrations, i.e., 0.1, 0.2,
0.5 and 1 mg/ mL, is showed in (Table-2) along
with experimental data on the drugs used as
standards, albendazole and Piperazine citrate. A
closer report of the data from the table indicates
that compounds 6b, 6c, 6d, and 6e have higher
activity than other compounds, which was confirmed
by their paralysis time of 0.46, 0.24, 0.56 and 0.26
min, respectively, and death time of 1.57, 0.39, 1.28
and 1.07 min, respectively, at a concentration of 1
mg/ mL. Compounds with presence of chlorine and
nitro substituent at the ortho and para position of
the phenyl ring 6c and 6e led to increase in activity.
Chlorine at ortho position of the phenyl ring provided
activity to compound 6b.The triazolyl ring clubbed
with benzimidazole moiety in same molecule frame
led to response in anthelmintic activity.
The starting product 2-mercapto
Benzimidazole prepared from o-phenylenediamine
and potassium ethyl xanthate. The 2-mercapto
benzimidazole was converted into the corresponding
ehylchloroacetate and potassium carbonate which
on esterification afforded ethyl 2-(1H-benzo[d]
imidazol-2-ylthio)acetate (2). This in turn on
treatment with hydrazine hydrate gave 2-(1H-
benzo[d]imidazol-2-ylthio) acetohydrazide (3)
and was reacted with carbon disulphide in the
presence of alcoholic potassium hydroxide to
give of potassium 2-(2-(1H-benzo[d]imidazol-2-
ylthio) acetyl) hydrazinecarbodithioate (4). The
finally product 4 and different aryloxyacetic acid
hydrazides (5) were subjected to reflux and followed
by acidification gave the product, which was purified
by recrystallization from ethanol and the compounds
were characterized as N-[3-{(1H-benzo[d]imidazol-2-
ylthio) methyl}-5-mercapto-4H-1, 2, 4-triazol-4-yl]-2-
substituted phenoxy acetamide (6a-g).In IR Spectra
6a-6g has been found to exhibit characteristic
absorption bands (in cm-1) at :3150(NH) 3050-2910
(C-H str., aromatic and aliphatic) 2550 (SH) or 1150
C=S, 1660 (CO of CONH), 1605 (C=N), 1500 (C-N),
845(Substituted benzene),1H NMR spectrum of the
compound (in CDCl3) has showed characteristic
proton signals (in δ, ppm) at : 2.3(s.3H,CH3),
4.6(s,2H,(OCH2)), 4.2(d,2H,SCH2),8.4(d,1H, NH),
7.0-7.4(m,9H,Ar-H),10.35(s,1H,NH),peak for SH
not observed. Based on the spectral data, the
compound 6g was characterized as N-[3-{(1H-
benzo[d]imidazol-2-ylthio) methyl}-5-mercapto-4H-1,
2, 4-triazol-4-yl]-2-(p-tolyloxy) acetamide and the
CONCLUSION
Present research work involves synthesis
of some benzimidazole derivatives clubbed with
triazolyl moiety and to explore their anthelminthetic
activity.The compounds (6d, 6e) have been exhibited
good anhelminthetic activity against earth worm due
to presence triazolyl moiety with phenyl substituted
electron withdrawing groups and conjugated with
pharmacophore group such as benzimidazole ring
in the same molecular structural frame.
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