IAJPS 2018, 05 (01), 706-711 Monika et al ISSN 2349-7750

CODEN [USA]: IAJPBB ISSN: 2349-7750

INDO AMERICAN JOURNAL OF

PHARMACEUTICAL SCIENCES
http://doi.org/10.5281/zenodo.1174182

Available online at: http://www.iajps.com Research Article

SYNTHESIS AND ANTIMICROBIAL ACTIVITIES OF NOVEL

BENZIMIDAZOLE DERIVATIVES
Monika1* , Ramanpreet Kaur1, Ravinder Sharma1 , Gunpreet Kaur2, Harjinder Singh3
1University Institute of Pharmaceutical Sciences and Research, Faridkot, India
2University Centre of Excellence in Research, BFUHS, Faridkot, India
3
Civil Hospital, Faridkot, India
Abstract:
Benzimidazole is a heterocyclic aromatic organic compound. Benzimidazole derivatives are of wide interest because
of their diverse biological activity and clinical applications, they are remarkably effective compounds both with
respect to their inhibitory activity and their favorable selectivity ratio. Looking at the importance of benzimidazole
and oxadiazole nucleus, it was thought that it would be worthwhile to design and synthesize new benzimidazole
derivatives. In recent years, attention has increasingly been given to the synthesis of benzimidazole derivatives.
Hence, there will always be a vital need to discover new chemotherapeutic agents to overcome the emergence of
resistance and ideally shorten the duration of therapy. Due to the structural similarity to purine, antibacterial
ability of benzimidazoles is explained by their competition with purines resulting in inhibition of the synthesis of
bacterial nucleic acids and proteins. Thus, from the present study it can be concluded that cyclohexyl derivatives of
benzimidazole ring have significant antibacterial activity.
Keywords: Benzimidazoles, anti-malarial, anti-fungal, anti-bacterial, anti-viral, anti biotics.
*Corresponding Author:
Monika, QR code
Assistant Professor,
Pharmaceutical Chemistry,
University Institute of Pharmaceutical Research,
Baba Farid University of Health Sciences,
Faridkot, India.
E-mail: [email protected]

Please cite this article in press as Monika et al., Synthesis and Antimicrobial Activities of Novel Benzimidazole
Derivatives, Indo Am. J. P. Sci, 2018; 05(01).

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 IAJPS 2018, 05 (01), 706-711 Monika et al ISSN 2349-7750

INTRODUCTION: Recrystallization was carried out using dry solvents.

Benzimidazoles are regarded as a promising class of Charcoal treatment was given to purify compounds.
bioactive heterocyclic compounds that exhibit a Column Chromatography was carried out using silica
range of biological activities. Specifically, this gel (60-120 mesh) using Chloroform as solvent.
nucleus is a constituent of vitamin-B12 [1]. This ring
system is present in numerous antioxidant [2,3], Spectral characterization:
1
antiparasitic [4,5], antihelmintics [6], antiproliferative H-NMR and 13C-NMR Spectra were recorded on a
[7], anti- HIV [8], anti-inflammatory [9-10] and 300 MHz FT-NMR and 75.45 MHz respectively. In
antineoplastic [11,12] activities. Our research efforts 1
H-NMR chemical shifts were reported in δ values
are directed to find new chemical classes of using tetramethylsilane as internal standard with
antimycobacterially active agents. A lot of studies number of protons, multiplicities (s-singlet, d-
were carried out on heterocyclic systems bearing an doublet, t-triplet, q-quartet, m-multiplet, dd-double
alkylsulfanyl group as pharmacophore. Varied doublet) and coupling constants (J) in Hz (Hertz) in
bioactivities exhibited by benzimidazoles, efforts the solvent indicated.IR spectra were recorded as
have been made from time to time to generate KBr pellets on Shimadzu 8400 S
libraries of these compounds and screened them for FT-IR Spectrophotometer and intensities mentioned
potential biological activities. This ring system is as s-strong, m-medium and w-weak.
present in numerous antiparasitic, fungicidal,
anithelemintic and anti-inflammatory drugs. The Material
earliest report of their antibacterial activity appeared All the chemicals used for preparing reagents were
in 1964 [13], and more recently we have found two procured from Hi-media and were of analytical
groups of substituted benzimidazoles, namely the 5, grade. All the glassware’s like test tubes, beaker and
6-dinitro and 2- trifluoromethyl derivatives, to be Erlenmeyer’s flask were of Borosil grade.
promising candidates for antimicrobial drugs [14]. In
this paper author present new data on the Test Organism
antimicrobial activities of derivatives of Microbial strains used were procured from Institute
benzimidazole ring. Various functional groups, such of Microbial Technology, Chandigarh, India. The
as, chloro, fluoro, nitro, methoxy, and so on, have an stock cultures were maintained on nutrient agar
important significance in medicinal chemistry. slants.
Especially the dihalogen-like dichloro, as in
antifungal azoles, provides a potent antifungal Preparation of Test Substance: The tested
activity for drugs such as oxiconazole, ketoconazole, compounds and standard were dissolved in dimethyl
terconazole, itraconazole and so on. sulfoxide (DMSO) using 1000µg/ml concentration.
DMSO was used as negative control and ampicillin
MATERIAL AND METHODS: was used as positive control.
Chemistry: General procedures Preparation of Mc Farland Standard: 0.5 Mc
followed during synthesis are as equivalent turbidity was prepared by using standard
follow: method.
Reaction Monitoring: Antibacterial Activity Assay: The cup plate or
Synthetic procedures employed were monitored by cylindrical plate method was applied to determine the
Thin Layer Chromatography (TLC) employing 6'’ x growth inhibition of bacteria by compounds to be
2” plates coated with 0.25 mm thick layer of silica tested. In this method, petri plates were prepared by
gel G. Solvent systems of chloroform: methanol pouring about 25ml of Muller Hinton Agar in each
mixtures (5:1) were used to monitor the reactions. plate for Escherichia coli, Salmonella typhi,
The dried plates after development were visualized in Staphylococcus aureus and Bacillus subtilis
an iodine chamber. respectively. One plate each for standards DMSO and
ampicillin were prepared. Then, 100µl of
Product separation: standardized inoculum suspension was poured,
Desired products were extracted with suitable solvent spreaded uniformly on respective agar plates and
(ether). Crushed ice was used instead of cold water dried for 5 minutes. 1µl test substance was poured in
wherever required in very hot days. Solvents were the uniform wells created in the centre of the
dried using anhydrous magnesium sulphate, fused petriplates and then the plates were kept undisturbed
calcium chloride, or specifically reported material. for 5-10 minutes followed by incubation at 370C for
24hrs. After incubation the diameter of circular area
Purification procedures: around the well known as Zone of Inhibition (ZOI)

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 IAJPS 2018, 05 (01), 706-711 Monika et al ISSN 2349-7750

was measured which is directly proportional to the Bacteria were cultivated at 37°C in Muller Hinton
sensitivity of test organisms to the test substance. Broth (MHB) and Muller Hinton agar (MHA). The
tested compounds and standard were dissolved in
Minimum inhibitory concentration (MIC): MIC is dimethyl sulfoxide (DMSO) using 1000 µg /ml
the concentration required to inhibit bacterial cell concentration. DMSO was used as negative control
proliferation by 50% after exposure of cells to the test and ampicillin (standard) was used as positive
compounds. MIC values (ug/ml) were determined control.
using turbidimetry method. In this method, test and
standards both were serially diluted with different Antibacterial activity
concentration (500, 250, 125 67 ug/ml) in Muller The cup plate method was applied to determine the
Hinton Broth (MHA). After inoculation, the tubes growth inhibition of bacteria by compounds to be
were incubated at 37°C for 24 hrs. Cell growth was tested. This method depends upon the diffusion of
determined by visible turbidity, in order to evaluate test substance from vertical cavity through solidified
the results. The lowest compound concentration, agar layer on petriplate. The growth of test
showing no visible growths after incubation were microorganism is inhibited entirely around the
expressed as Minimum inhibitory concentration circular area or the cavity containing test substance
(MIC) in µg/ml. which is measured after the incubation for 24hrs at
37°C.
Biological activity
Bacteria and media Determination of MIC
In vitro antibacterial studies of various synthesized For minimum inhibitory concentration, tested and
compounds were carried out on gram negative and standard compounds were serially diluted with
gram positive bacterial strains. The gram negative different concentration in Muller Hinton broth
strains are Escherichia. Coli (MTCC 1678, aerobic), (MHB) at pH 7.4. The dilutions were inoculated with
Salmonella typhi (MTCC 733, aerobic) and gram a suspension of bacterial culture in Muller Hinton
positive strains are Staphylococcus aureus (MTCC Agar (MHA). After inoculation, tubes were
3160, aerobic), Bacillus subtillis (MTCC 441, p incubated at 37°C for 24 hrs. Cell growth was
aerobic), procured from Microbial Type Culture determined by visible turbidity, in order to evaluate
Collection and gene bank (MTCC ), IMTECH the results. The lowest compounds concentration,
Chandigarh. showing no visible growths after incubation were
expressed as minimum inhibitory concentration
(MIC) in µg/ml.
Synthetic Scheme
Cl NHR or Ar

+ R or ArNH2

CH3 NO2 CH3 NO2

+
Zn

NHR or Ar
NR or Ar

SH CS2 +

CH3 N CH3 NH2

R or ArX

NR or Ar

SR or Ar

CH3 N

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 IAJPS 2018, 05 (01), 706-711 Monika et al ISSN 2349-7750

Spectral analysis of compounds [2] C13H18N2S:- IR Specturm: 𝛿 max(KBr): 2850-

1] C16H16N2S,Yield M.P 82-83°C Solubility: - 2960 cm-1 (C- Hstr aliohatic) ,1487 (C−N), 788 cm-
1
Chloroform and Ethanol (C-S)
Spectral analysis: - IR Specturm (KBr):1450cm-1 NMR Specturm: 1H NMR (300 MHz, CDCl3): 𝛿H 7.4
(C−N), 790 cm-1(C-S) (s,-1H,-Ph) 𝛿H 7.0 (d,-1H,-Ph) 𝛿H 6.9 (d,-1H,-Ph) 𝛿H
NMR Specturm 1H NMR (300 MHz, CDCl3): 𝛿H2.46 3.6(s,-3H,-CH3) 𝛿H 3.3(t,-2H,-CH2) 𝛿H 2.4(s,-3H,-
(s,-3H,-CH3 ) 𝛿H3.45(s,-3H,-CH3 ) 𝛿H3.54(s,-3H,- CH3) 𝛿H 1.7(m,-2H,
CH3) 𝛿H4.56(s,-2H,-CH2) 𝛿H7.05(q,-1H,-Ph) CH2) 𝛿H 1.4 (m,-2H,-CH2) 𝛿H 0.9 (t,-3H,-CH3)
𝛿H7.33(m,-5H,-Ph) 7.38(dd,-1H,-Ph) : 𝛿H 7.50 (s,- 13
CNMR(75.45MHz,CDCl3):13.5,21.4,21.7,29.9,31.3
1H,-Ph)).13C NMR (75.45 MHz, CDCl3):24.34, ,32.3,107.8,117.9,123.1,131.4,134.5,143.1,152.0 and
31.26, 112.56, 115.46, 125.82, M+234
127.23,127.85,128.82,131.32,132.71, 138.83,139.77,
152.61 M+268

BIOLOGICAL EVALUATION
The synthesized compounds were evaluated for antibacterial and antimycobacterial activity. Ampicillin was used as
standard for antibacterial activity.
Serial No. Code X Y
1. HA-1 CH3 CH2-C6H5
2. HA-2 CH3 C4H9
3. HA-3 C2H5 CH2-C6H5
4. HA-4 C2H5 C4H9
5. HA-5 C3H7 CH2-C6H5
6. HA-6 C3H7 C4H9
7. HA-7 C4H9 CH2-C6H5
8. HA-8 C4H9 C4H9
9. HA-9 CH2-C6H5 CH2-C6H5
10. HA-10 CH2-C6H5 C4H9
11. HA-11 C6H11 CH2-C6H5
12. HA-12 C6H11 C4H9

N
S Y

CH3 N X- R or Ar
Y- R or Ar
ANTIBACTERIAL ACTIVITY
The compounds were screened for their antibacterial activity against different bacterial strains (E.coli, S.tyhpi,
B.subtilis, S.aureus) using concentration of 1000µg/ml in Dimethyl sulfoxide solution and evaluated by cup-plate or
cylindrical plate method. The compounds were assigned codes by HA-1-12. The results obtained are shown in Table
1. It has been found that good activity was exhibited by HA-8 and HA-12 against all bacterial strains.

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 IAJPS 2018, 05 (01), 706-711 Monika et al ISSN 2349-7750

Table 1: In vitro zone of inhibition of various bacterial strains by benzimidazole (HA1-HA12) derivatives
Growth inhibition zone diameter (mm)
Compound E.coli S.tyhpi B.subtilis S.aureus
code (1678) (733) (441) (3160)
HA-1 - - 3 6
HA-2 - - 5 6
HA-3 - - 4 5
HA-4 - - 4 7
HA-5 - - 3 6
HA-6 - - - 4
HA-7 - 3 - 5
HA-8 5 8 5 8
HA-9 - 4 - -
HA-10 - - - -
HA-11 - 3 - -
HA-12 6 7 4 9
Ampicillin 11 14 10 12

RESULTS AND DISCUSSION: HA-9 and HA-11 having aromatic group at 1-position
The in vitro evaluation of the synthesized compound and benzyl at 2-position shows inhibition against
against four different bacterial strains i.e E.coli, only one gram negative strain i.e S.typhi and no
S.typhi, B.subtilis, S.aureus have shown moderate to inhibition against all other strains.
significant activity. Compound HA-1,2,3,4 and 5
shows less zone of inhibition ranges between 3mm Further MIC values of these two compounds lies
to 7mm against gram positive strains i.e. B.subtilis within the range of less than 125µg/ml and greater
and S.aureus as compared to Ampicillin (standard) than 500µg/ml. Thus, it can be found that mostly
but no activity against gram negative strains i.e. compounds having aliphatic group at 1-position of
E.coli and S.typhi. Compound HA-6 shows mild benzimidazole ring shows activity against gram
activity against S.aureus but no activity against all positive strains i.e. B.subtilis and S.aureus.
other strains. Compound HA-7 shows zone of Compound HA-12 shows MIC value less than
inhibition against one gram negative bacteria i.e. 125µg/ml against E.coli strain and less than 250
S.typhi and one gram positive bacteria i.e. S.aureus µg/ml against S.aureus and less than 500µg/ml
and no activity against E.coli and B.subtilis. against S.typhi [16]. Compound HA-8 shows
Compound HA-8 and HA-12 shows good zone of moderate MIC value greater than 500µg/ml against
inhibition against all strains [15]. Compound HA-10 all strains (Table 2).
shows no inhibition against all strains. Compound

Table 2: In vitro antibacterial activity of compound HA-8 and HA-12 against E.coli, S.typhi, B.subtilis,
S.aureus (MIC expressed in µg/ml)
Concentrati 67 µg/ml 125 µg/ml 250 µg/ml 500 µg/ml
on
Strains HA HA Ampicill HA HA- Ampicill HA HA- Ampicill HA- HA- Ampicill
-8 -12 in -8 12 in -8 12 in 8 12 in
E.coli - - >67 - >12 - - + - <50 + -
5 0
S.typhi - - >67 - - - - - - <50 >50 -
0 0
B.subtilis - - >67 - - - - - - <50 <50 -
0 0
S.aureus - - >67 - - - - >25 - <50 + -
0 0

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CONCLUSION: 8.Rao A, Chimmiri A, De Clercq E, Monforte AM,

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concluded that cyclohexyl derivatives of 10.Lackner TE, Clissold SP. A review of its
benzimidazole ring have significant antibacterial antimicrobial activity and therapeutic use in
activity. superficial mycoses. Drugs, 1989; 38: 204-225.
11.Monem A, Hafez A. Benzimidazole condensed
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