Asian Journal of Chemistry Vol. 21, No.

5 (2009), 3367-3370

Synthesis and Antifungal Activity of Isatin-3-semicarbazone

S.N. PANDEYA†, KISHU TRIPATHI* and SHOBHA KULSHRESHTHA‡

Department of Pharmaceutical Chemistry, Surya College of Pharmacy, Lucknow-227 305, India
E-mail: kishutripathi@rediffmail.com

Isatin-3-semicarbazone and substituted isatin-3-semicarbazone were

synthesized and their antifungal activity was screened. It was observed
that the semicarbazones showed inhibitory activity at 100-500 µg/mL.

Key Words: Isatin-3-semicarbazone, Antibacterial activity.

INTRODUCTION
Since the 1970's, there has been a steady increase in the incidence of serious
secondary systemic fungal infections. One of the factor aiding the spread of fungal
disease has been the wide spread use of broad-spectrum antibiotics, which elimi-
nate or decrease the non-pathogenic bacterial populations that normally compete
with fungi. Another has been the increased number of individuals with reduced
immune responses caused by the acquired immuno-deficiency syndrome (AIDS)
or by the action of immunosuppresant drugs or cancer chemotherapy agents. This
has led to an increased prevalance of opportunistic infections i.e., infections with
fungi that rarely cause disease in healthy individuals.
Majority in the World, the commonest systemic fungal infections are blasto-
mycosis, histoplasmosis, coccidiomycosis and para-coccidiomycosis; these are
primary infections i.e., they are not secondary to reduced immunological function
or altered commensal microorganisms1-3.
The need for better clinical agents has emerged because of the increasing de-
tection of systemic mycosis in patients suffering from debilitating diseases such as
in neoplasias. Systemic mycosis has been found in 61 % of patient dying with
acute leukemia, in 45 % deaths in renal transplant recipients and 75 % of AIDS
patients4. The compounds synthesized in this study with isatin moiety could very
well be new chemical entities as antifungal agents and so the compounds were
tested for antifungal activity5,6.
Isatin, also named indoline-2, 3-dione, is a bright coloured compound with a
long history and a broad range of pharmacological actions. Chemically, isatin may
be characterised as lactam of o-amino-benzoylformic acid. It possesses both amide
group and a keto group.

†Saroj Institute of Technology & Management, Lucknow-226 021, India.

‡Department of Pharmacolgy, S.N. Medical College, Agra-282 002, India.
3368 Pandeya et al. Asian J. Chem.

EXPERIMENTAL
Melting points were taken by Open Capillary method and are uncorrected. UV
Spectra were recorded on a Jasco 7800UV/Visible spectrophotometer and IR spectra
on a Jasco 5800 FT-IR using KBr discs.The 1H NMR spectra were recorded on a
Jeol FX-90Q spectrophotometer in DMSO-d6 or CDCl3 with TMS as an internal
standard. Elemental analysis were determined with Perkin-Elmer model 240
analyzer. The purity of the compounds was confirmed by thin layer chromatography
using silica gel plates and different solvent systems. Isatin was purchased from
Ward, Blenkinsop & Co. Ltd.
A mixture of semicarbazide hydrochloride (0.1 mol, 11.1 g) and sodium
acetate (0.15 mol, 12.3 g) was dissolved in 100 mL of water. To this isatin (0.1 mol,
14.7 g) dissolved in 150 mL of alcohol was added and the whole mixture stirred for
10-15 min. It was warmed on a water-bath to dissolve the contents and then cooled
in ice when a yellow precipitate was obtained (Scheme-I). It was filtered, dried and
recrystallized with alcohol, m.p. 270 °C, yield 61.0 %. Similarly, 5-subsituted isatin-
3-semicarbazones were prepared from corresponding isatin (Table-1).
Spectral data: UV (λmax MeOH): 400; IR (KBr, νmax, cm-1): 3550-3400s (N-H),
1730s (CONH2), 1710m (C=O), 1690m (amide C=O), 1680s (C=N); 1H NMR
(DMSO-d6, δ ppm): 7.0 (s, 2H, CONH2) 7.2-7.5 (m, 4H, aromatic), 8.2 (d, 1H, NH,
indole), 10.5 (s, 1H, =NNH D2O exchangeable).
R
R NH2OH.HCl
OH
Cl3CCH(OH)2
NH2 N
Na2SO4
NH
Substituted Aniline
O

Isonitrosoacetanilide

conc. H2SO 4

H2N
O
O
N NH H2NNHCONH2.HCl R
R
O
O N
N Alcohol,CH3COONa H
H
Isatin
Isatin-3-semicarbazone
R =H, 5-CH3, 5-Cl, 5-Br, 5-NO2, 5-OCH3
(II-1- II-6)

Scheme-I: Synthesis of isatin-3-semicarbazone (II-1) and substituted

isatin-3-semicarbazones (II-2-6)
Vol. 21, No. 5 (2009) Synthesis and Antifungal Activity of Isatin-3-semicarbazone 3369

TABLE-1
CHARACTERIZATION DATA OF 5-SUBSTITUTED ISATIN-3-SEMICARBAZONES
Compd. No. m.p. (ºC) / Analysis %
R Rf value IR (KBr, νmax, cm-1)
(m.f.) Yield (%) Calcd. Found
C: 45.29 C: 45.35 3550-3400m (NH stretch),
II-2 210
5-Cl H: 2.95 H: 2.68 0.71 1720s (C=O), 1680s (C=N),
(C9H7ClN4O2) (62)
N: 23.48 N: 23.32 550m (C-Cl)
C: 56.03 C: 55.42
II-3 245 3400s (NH stretch), 1720s
5-CH3 H: 4.61 H: 4.83 0.73
(C10H10N4O2) (69) (C=O), 1685m (C=N)
N: 25.68 N: 25.74
C: 38.18 C: 38.23 3450-3400m (NH stretch),
II-4 205
5-Br H: 2.49 H: 2.31 0.68 1750s (C=O), 1680s (C=N),
(C9H7BrN4O2) (68)
N: 19.79 N: 19.83 1400m (aromatic)
C: 43.37 C: 43.62 3420, 3340, 1725m (C=O),
II-5 160
5-NO2 H: 2.83 H: 2.57 0.38 1680s (C=N), 1500s
(C9H7N5O4) (67)
N: 28.11 N: 28.28 (aromatic)
C: 51.27 C: 51.53 3400s (NH stretch), 1720s
II-6 215
5-OCH3 H: 4.30 H: 4.51 0.62 (C=O), 1670m (C=N),
(C10H10N4O3) (54)
N: 23.92 N: 23.85 1570s (aromatic)

Antifungal activity6-8
Culture media: Sabouraud glucose agar (SGA) medium was used. Glucose
(4 g), agar agar (2.0 g) and peptone (1.0 g) were dissolved in 100 mL of distilled
water and pH was adjusted to 6-8 using acid or alkali. The medium was sterilized in
an autoclave at 15 lbs per sq. inch for 15 min.
Preparation of test samples: Each test sample (0.5 mg) was dissolved in 5 mL
of DMF (100 mg/mL) with warming to form the stock solution. From each stock
sample, 1 mL was poured into each sterile test tube.
Tube slant culture method: Sabouraud glucose agar (SGA) medium was
melted, cooled to 45-50 ºC and poured 1 mL each into test tubes containing 1 mL
of test samples to make the final concentration of test samples 100 mg/mL. The
contents were mixed well without formation of any air bubbles. The cotton plugged
test tubes were then kept at an angle of 30 ºC and media were allowed to solidify to
form slants.
In the aseptic UV chamber, the slants were inoculated with a platinum loopful
of the fungi from stock culture so that fungi were just embedded into the slant and
also spread over the surface of the slant. The inoculated slants were then in incubated
at 37 ºC in an incubator and growth of fungi studied after 45 h and 7 d. The growth
of fungi in the slant was compared for the test samples against the control group7,8.
RESULTS AND DISCUSSION
The antifungal activity of isatin-3-semicarbazones (II-1 to II-6) were investigated
against Candida albicans and Aspergillus niger at different concentrations and the
semicarbazones showed inhibitory activity at 100-500 µg/mL. The antifungal activity
3370 Pandeya et al. Asian J. Chem.

exhibited by these compounds is not very encouraging. The isatin-3-semicarbazone

(II-1) was found to be active at the concentration used. Among the semicarbazones,
the 5-chloro (II-2) and 5-bromo (II-4) showed a zone of inhibition of 1.6 and 2.2 cm
(Table-2).
TABLE-2
ANTIFUNGAL ACTIVITY OF ISATIN-3-SEMICARBAZONES
Bacteria II-I II-2 II-3 II-4 II-5 II-6
Candida albican + + – + – –
Aspergillus niger + – – – – –
Concentration = 100 mg/mL for II-1; 500 mg/mL for II-2 to II-6
+ = Inhibition; – = no inhibition; ± = Weak inhibition.

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(Received: 11 February 2008; Accepted: 5 February 2009) AJC-7200

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