Research article Rapports De Pharmacie Vol.

2 (4), 2016, 315-318

ISSN: 2455-0507
SYNTHESIS ANTIBACTERIAL AND ANTIFUNGAL EVALUATION OF
NOVEL ARYLAZO PYRAZOLE DERIVATIVES
B. C. Revanasiddappa*, M. Vijay Kumar, Prashanth Nayak, Neethu Jose,
Ajmal Roshan Ali, Apoorva, Yashaswini
*Department of Pharmaceutical Chemistry, NGSM Institute of Pharmaceutical
Sciences of Nitte University, Paneer, Deralakatte, Mangalore-575 018,
Karnataka, India
ABSTRACT
A novel series of substituted aryl Azopyrazoles (3a-j) were synthesized by reacting various oxobutyrates (2a-
j) with 2-hydrazino benzothiazole (1) in glacial acetic acid medium. The key intermediate compounds (2a-j)
were prepared by diazotization of appropriately substituted anilines followed by condensation with acetyl
acetone in alcohol medium. The newly synthesized compounds were assigned on the basis of IR, 1H-NMR
and Mass spectral data. All the newly synthesized compounds were evaluated for their In-Vitro antibacterial
and antifungal activities
Keywords: Pyrazoles, Arylazo pyrazoles, Diazotization, Antibacterial activity, Antifungal activity
INTRODUCTION
Since the discovery of Antipyrine as analgesic and dyeing of textile fibers, coloring of different
antipyretic drug, Pyrazole ring has attracted much materials, biomedical studies, and advanced
attention and a large number of Pyrazole derivatives applications in organic synthesis [7-10].
have been synthesized as potential antibiotics, In the present work, we have planned to synthesize a
antidiabetic, antineoplastics. Pyrazole chemically new series of aryl azo Pyrazoles, since the later
known as 1, 2-diazole has become a popular topic compounds is found to be an one of the important
due to its manifold uses. The Pyrazole ring is a class of heterocyclic compounds, because of their
prominent structure motif found in numerous varied biological and pharmacological exhibited by
pharmaceutically active compounds. This mainly these compounds.
due to the easy preparation and important biological
MATERIALS AND METHODS
activity. Pyrazole framework plays an essential role
Experimental:
in biologically active compounds and therefore
IR spectra were recorded by using Alpha Bruker IR
represents an interesting template for combinatorial
Spectrometer in KBr disc (cm-1).1H-NMR spectra
as well as medicinal chemistry.
were recorded on Bruker Avance II 400 NMR
Pyrazoles are associated with various biological
Spectrometer using TMS as internal standard. All
activities like antimicrobial[1], antineoplastic[2],
the melting points were determined by open
analgesic[3], anti-inflammatory[4], antifungal[5],
6 capillary method and are uncorrected. Thin layer
MAP kinase inhibitory , monoamine oxidase
chromatography was used for monitoring the
inhibitory activity[6], anti-HIV[6] etc.,
reaction and to check the purity. (Silica gel G plates
Similarly compounds having azo or hydrazano (Merck), Solvent system: ethyl acetate: pet. ether).
groups exhibit a wide varieties of biological Chemical shift values were reported as values in
activities. Literature survey reveals the importance ppm relative to TMS (=0) as internal standard.
of diazo substitution on various heterocyclic Mass spectra were recorded on ESI.
moieties in enhancing the biological activity. Azo Antimicrobial activity:
compounds are of great importance due to their All the newly synthesized compounds (3a-j) were
versatile application in various fields, such as the screened for their antifungal and antibacterial
activities. Four bacterial species were chosen for the
Address for correspondence: study of antibacterial activity, two Gram Negative
Dr. B.C. Revanasiddappa strains (E.coli, P.aeruginosa) and two Gram Positive
Dept. of Pharmaceutical Chemistry strains (B.subtilis, S.aureus). The antibacterial
NGSM Institute of Pharmaceutical Sciences activity was conducted using Cup-Plate agar
Paneer, Deralakatte, Mangalore-575 018, diffusion method[11]. The fungicidal activity of the
Karnataka. compounds was evaluated against A.flavus and
Email: evergreen_revan@rediffmail.com A.fumigatus. All the synthesized compounds were

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 Revanasiddappa et al,: Synthesis antibacterial and antifungal evaluation of arylazo pyrazole derivatives

tested at a conc. of 100 g/ml. Ciprofloxacin and
Flucanazole were used as standard drugs for
comparison of antibacterial and antifungal activity
respectively. The activities were determined by
measuring the diameter for the inhibition of zone in
mm. The antimicrobial data of the compounds (3a-j)
is given in table 2.
In the antibacterial activity all the tested compounds
showed very weak to moderate activity against all
the four pathogenic micro-organisms. None of the
tested compounds was found to potent when
compared to the standard Ciprofloxacin.
In the antifungal activity, all the tested compounds
showed weak to moderate activity against both
fungal organisms. Most of the tested compounds
showed very weak activity against A.fumigatus.
Synthesis of oxobutyrates (2a-j):[12]
Aniline (9.3 ml, 0.1 mol) was dissolved in aqueous
hydrochloric acid (80 ml, 1:1). The contents were
stirred, cooled (0-2C) and cold solution of sodium
nitrite (12.0 g in 30 ml water) was slowly added
maintaining the temperature between 0-2C. The
cold diazotized solution was added drop wise with
stirring to a well cooled mixture of acetyl acetone
(0.1 mol, 10 ml) and sodium acetate (12 g dissolved
in 10 ml of 50% aqueous ethanol). Stirring was
further continued for forty five minutes, when
yellow crystals separated. The product was filtered
under suction, washed with water and recrystallized
from aqueous ethanol.
Synthesis of Aryl azo Pyrazole Derivatives (3a-j):
A mixture Oxobutyrates (2a-j) and 2-hydrazino
benzothiazole (0.01 mol) (1) were dissolved in 30 ml
of glacial acetic acid. The reaction mixture was
refluxed for about 16-22 hrs and it is cooled. The
contents of the reaction mixture were poured into the
crushed ice. The resulted precipitated compound is
filtered and recrystallized from suitable solvents.
The physical data of compounds (3a-j) is given in
table 1.

3a:IR(KBr,cm-1): 1530 (N=N), 1588(C=C), 1631(C=N), 3128(C-H).1H-NMR (CDCl3,): 2.49 (s,

CH3, 3H), 2.69 (s, CH3, 3H), 7.16-7.95 (m, Ar-H, 9H). MS(m/z): 333(M+).
3d:IR(KBr,cm-1): 1535 (N=N), 1550(C=C), 1593(C=N), 2921(C-H).1H-NMR (CDCl3,): 2.58 (s,
CH3, 3H), 3.19 (s, CH3, 3H), 7.36-7.95 (m, Ar-H, 8H). MS(m/z): 351(M+)
3f:IR(KBr,cm-1): 1537 (N=N), 1565 (C=C), 1598(C=N), 2950(C-H).1H-NMR (CDCl3, ): 2.62 (s,
CH3, 3H), 3.17 (s, CH3, 3H), 7.15-7.94 (m, Ar-H, 8H). MS (m/z): 369(M+1).
Table-1: Physical data of Arylazo Pyrazoles (3a-j)

Comp R-NH2 Mol M.P Yield

Wt (0C) (%)

3a C6H5 333 108-110 61

3b 4-NO2 378 124-126 60
3c 4-Br 412 115-117 60
3d 2-F 351 146-148 58
3e 2-Cl 368 92-94 57
3f 4-Cl 368 172-174 67
3g 2-CH3 347 188-190 68
3h 4-CH3 347 157-159 67
3i 4-(CH3)2 361 131-133 65
3j 3-NO2 378 137-139 68

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 Revanasiddappa et al,: Synthesis antibacterial and antifungal evaluation of arylazo pyrazole derivatives

Table 2: Antimicrobial activity of Compounds (3a-j)

Diameter of zone of inhibition (mm)

Comp.
S.aureus B.subtilis E.coli P.aeruginosa A.flavus A.fumigatus

3a 07 11 11 09 11 11
3b 08 10 12 08 10 10
3c 09 11 10 07 11 08
3d 10 10 10 08 12 08
3e 10 12 11 09 11 10
3f 09 10 12 10 10 09
3g 08 09 10 09 11 08
3h 09 08 11 10 10 08
3i 10 09 09 11 12 08
3j 10 10 08 10 10 08
Ciprofloxacin 25 23 24 24 - -
Fluconazole - - - - 23 23

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 Revanasiddappa et al,: Synthesis antibacterial and antifungal evaluation of arylazo pyrazole derivatives
RESULTS AND DISCUSSION
In the present work, we report herein a novel series
of arylazo pyrazole derivatives were synthesized and
evaluated for their antibacterial activities.
Oxobutyrates (2a-j) were in turn prepared by
starting from different substituent anilines. The
suitable anilines were converted into diazonium salts
and then treated with acetyl acetone in presence of
sodium acetate to get corresponding Oxobutyrates
(2a-j). 2-hydrazino benzothiazole (1) reacts with
Oxobutyrates (2a-j) to yield the title compounds
substituted aryl azo pyrazoles derivatives (3a-j) in
excellent yield.
In compound 3a, C-H stretching was observed at
3218 cm-1. The C=N and N=N stretching was
observed at 1631cm-1 and 1530 cm-1 respectively. In
the 1H-NMR spectrum of compound 3a, the two
methyl protons resonated as singlet at 2.49 and
2.69 integrating for three protons respectively. The
aromatic protons were observed as multiplets in the
region of 7.16 to 7.95 integrating for nine protons.
Further evidence for the proposed structure was
obtained by recording mass spectra of the
synthesized compounds. The mass spectrum of the
compound 3a, showed the M+ peak at m/z 333
which is consistence with the molecular formula.
ACKNOWLEDGEMENTS
The authors are thankful to authorities of NGSM
Institute of Pharmaceutical Sciences, Nitte
University, Mangalore for providing all the
necessary facilities. The authors are thankful to
SAIF, Vellore Institute of Technology, Vellore for
providing the spectral data.
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