Synthesis and antimicrobial activity of o- and p-hydroxybenzoic acid thiosemicarbazides

Article abstract of DOI:10.1134/S107036321204010X

The ortho- and para-hydroxybenzoic acid thiosemicarbazide derivatives which are potentially biologically active substances were obtained by the reaction of the corresponding hydrazides with various isothiocyanates. Their structures were determined using I

Full text of DOI:10.1134/S107036321204010X

ISSN 1070-3632, Russian Journal of General Chemistry, 2012, Vol. 82, No. 4, pp. 668–671. © Pleiades Publishing, Ltd., 2012.
Original Russian Text © O.A. Nurkenov, Zh.B. Satpaeva, I.V. Kulakov, S.B. Akhmetova, S.K. Zhaugasheva, 2012, published in Zhurnal Obshchei Khimii,
2012, Vol. 82, No. 4, pp. 582–585.
Synthesis and Antimicrobial Activity
of o- and p-Hydroxybenzoic Acid Thiosemicarbazides
O. A. Nurkenova, Zh. B. Satpaevaa, I. V. Kulakova,
S. B. Akhmetovab, and S. K. Zhaugashevab
a Institute of Organic Synthesis and Coal Chemistry of Republic Kazakhstan,
ul. Alikhanova 1, Karaganda, 100008 Kazakhstan
e-mail: kulakov_iv@mail.ru
b Karaganda State Medical University, ul. Gogolya 40, Karaganda, 100008 Kazakhstan
Received February 14, 2011
Abstract—The ortho- and para-hydroxybenzoic acid thiosemicarbazide derivatives which are potentially
biologically active substances were obtained by the reaction of the corresponding hydrazides with various
1
isothiocyanates. Their structures were determined using IR, H NMR spectroscopy, and mass spectrometry.
Pronounced antimicrobial activity of synthesized derivatives was revealed.
DOI: 10.1134/S107036321204010X
The retrieval of chemical substances possessing
antibacterial activity is carried out now on the ground
of certain scientific principles and quantitative
approaches to predicting the structure of compounds,
and in essence their purposeful synthesis is performed.
In the development of research in this area several
trends can be traced, one of them is the introduction of
pharmacophore fragments ino the structure of the
desired molecule. Such fragments include the
hydrazide and thiosemicarbazide groups and many of
their derivatives [1–3]. It is known also that thio-
semicarbazide derivatives possess a wide range of
biological effects: anticonvulsant, hypoglycemic, anti-
inflammatory and antibacterial [4, 5]. One of the first
antiviral drugs with the thiosemicarbazide fragment
was tibon (the para-acetaminobenzaldehyde thioaceta-
zone or thiosemicarbazone) [5]. Thiosemicarbazide
derivatives are also common among the tuberculostatic
preparations [6–8].
II were obtained by hydrazinolysis of methyl salicylate
and nipagin (methyl o- and p-hydroxybenzoates).
The hydrazides addition to various isothiocyanates
is one of the convenient methods of the synthesis of
substituted thiosemicarbazides, which are of great
interest not only in terms of a possible study of
biological activity, but also as the starting compounds
for the synthesis on this ground of the pharma-
cologically important nitrogen-containing hetero-
cycles. For example, we have described [9] the syn-
thesis of salicylic N-2-vinyloxyethylthiosemicarba-
zide and a possibility of its cyclization to 1,2,4-tri-
azole-3(4H)-thione.
In continuation of our research and to expand the
range of new biologically active substances, we
performed a reaction of nucleophilic addition of o- and
p-hydroxybenzhydrazides I and II to allyl isothio-
cyanate. The reaction was carried out in alcohol at the
equimolar ratio of reagents.
In this regard, we were interested in the synthesis of
new thiosemicarbazide derivatives based on the
salycilic and p-hydroxybenzoic acid hydrazides, and in
the study of their antimicrobial activity. Special
attention deserve in this respect the salicylic acid
hydrazide derivatives which are widely used as anti-
pyretic, antirheumatic, anti-inflammatory, analgesic
and tubercocidal drugs [4, 5]. Initial hydrazides I and
Synthesized compounds III and IV, respectively,
are white crystalline substances, readily soluble in
polar organic solvents. The IR spectra of the
synthesized compounds contain an absorption band at
1330–1310 cm–1 characteristic of –NH–CS group of
the thiosemicarbazide fragment, the absorption bands
of the amide group C(O)NH appear at 1690–1675 cm–1,
and of NH group, at 3390–3360 cm–1.
668

SYNTHESIS AND ANTIMICROBIAL ACTIVITY
669
R2
R1
C
R2
R1
C
H2C=CHCH2N=C=S
O
O
S
β
2
4
1
5
3
NHNHCNHCH2CH=CH26a,6b
α
NHNH2
III, IV
I, II
R1 = OH, R2 = H (I, III); R1 = H, R2 = OH (II, IV).
1
In the H NMR spectrum of the p-hydroxybenzoic
acid allylthiosemicarbazide IV the signals of α and β
protons of the aromatic ring are in a weak field: Hα
doublet at 7.78 ppm [J(HαHβ) = 8.7 Hz] and Hβ doublet
at 6.81 ppm. Aromatic hydroxyl proton gives rise to a
singlet at 9.10 ppm. Amide and thioamide N–H
protons also give downfield signals: three singlets, at
6.10 (H1), 9.25 (H2) and 8.2 ppm (H3). The methine
proton H5 of vinyl fragment appears as a complex
multiplet at 5.82 ppm. The methylene protons H6a and
H6b of the same vinyl fragment are manifested by two
doublets at 4.5 and 5.14 ppm with the spin-spin coupl-
ing constant J(H6αH6β) = 17.27 Hz. Methylene protons
of the NCH2 resonate at 4.9 ppm as a broad triplet.
The reaction was performed in dilute hydrochloric
acid at 95°C for 4 h. Basic physical and chemical
characteristics and the elemental analysis data of
synthesized compounds III–VIII are listed in Table 1.
The IR spectra of o- and p-hydroxybenzoic acid
thiosemicarbazides V and VI contain absorption bands
of stretching vibrations of NH2 groups in the region of
3305–3240 cm–1. In the regions of 1660 cm–1 and
1270 cm–1 there are absorption bands of carbonyl
(C=O) and thiocarbonyl (C=S) groups, respectively.
1
The analysis of H NMR spectrum of compound V
reveals characteristic signals of the aromatic ring.
Thus, in a weak field the signals are present of
aromatic protons H1–H4: H1 doublet at 6.89 ppm, H2
triplet at 7.42 ppm, H3 doublet at 6.93 ppm, and H4
doublet at 7.81 ppm. Aromatic hydroxyl proton
resonates as a singlet at 9.42 ppm. Amide and
thioamide N–H protons give three singlets at 11.89
ppm (H5), 10.52 ppm (H6), and 7.9 ppm (H7).
In order to obtain new thiosemicarbazide deriva-
tives we synthesized monosubstituted thiosemicar-
bazide derivatives by the reaction of o- and p-hyd-
roxybenzhydrazides I and II with potassium thio-
cyanate.
1
R2
2
R1
As known, glucosyl isothiocyanates play an
important role in carbohydrate chemistry as synthons
for producing various biologically active compounds
[10]. The introduction of carbohydrate residues in the
structure of biologically active substances increases the
solubility of the latter in water and leads to a sharp
decrease in the toxicity [11, 12]. Therefore by the
HCl
O
I, II + KSCN
3
95oC
S
C
7
4
5
6
NHNHCNH2
VI, VI
R1 = OH, R2 = H (I, V); R1 = H, R2 = OH (II, VI).
Table 1. Physicochemical constants and elemental analysis data of the synthesized compounds III–VIII
Found, %
Calculated, %
H
Comp.
no.
Yield, %
mp, °С
Formula
C
H
N
C
N
85
53
84
52
96
57
213–215
215–216
217–218
219–220
136–137
145–146
52.84
52.90
45.67
45.86
48.95
49.11
5.72
5.80
4.55
4.49
5.43
5.45
16.96
17.12
20.12
19.98
7.95
C11H13N3O2S
C11H13N3O2S
C8H9N3O2S
52.57
52.57
45.49
45.49
48.79
48.79
5.21
16.72
16.72
19.89
19.89
7.76
III
5.21
IV
4.29
V
C8H9N3O2S
4.29
VI
C22H27N3O11S
C22H27N3O11S
5.03
VII
VIII
8.10
5.03
7.76
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 82 No. 4 2012

670
NURKENOV et al.
condensation of hydrazides I and II with 1-deoxy-
2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl isothiocyanate,
which was obtained in situ from tetra-O-acetyl-α-D-
glucopyranosyl bromide and lead thiocyanate, we
synthesized the corresponding acetylated glucosyl-
containing thiosemicarbazide derivatives of o- and p-
hydroxybenzoic acids (VII, VIII), potentially
possessing a biological activity.
O
S
CH2OAc
CH2OAc
R2
R1
C
R2
N=C=S
H
NCNHNHC
O
O
O
OAc
OAc
+
R1
VII, VIII
R1 = OH, R2 = H (I, VII); R1 = H, R2 = OH (II, VIII).
Ac
O
Ac
O
NHNH2
OAc
OAc
I, II
It is shown that glucosyl isothiocyanate readily
reacts with nucleophilic reagents and forms in a high
yield the N-substituted acetylated glucosyl-containing
thiosemicarbazide derivatives of o- and p-hydroxy-
benzoic acid VII and VIII, respectively. The syn-
thesized compounds are white crystalline substances,
soluble in polar organic solvents.
activity against gram-positive strains (Staphylococcus
aureus, Bacillus subtilis) (Table 2). Compounds IV, V,
and VII exhibit moderately strong antibacterial activity
only against gram-positive (Staphylococcus aureus,
Bacillus subtilis) strains. Compounds II and VII show
moderate activity against gram-negative strain of
Escherichia coli and yeast fungus Candida albicans.
Table 2 lists the numerical values of diameters of the
zones of growth inhibition of the test strains (mm).
In order to reveal the substances with pronounced
biological activity among the synthesized derivatives,
we carried out primary screening test of compounds
II–V, VII for antimicrobial activity against gram-
positive (Staphylococcus aureus, Bacillus subtilis) and
gram-negative (Pseudomonas aeruginosa, Escherichia
coli) strains by the agar diffusion method. The
reference preparations were gentamicin for bacteria
and nystatin for fungi. Antimicrobial activity of
compounds II–V, VII was estimated by measuring the
diameter (mm) of the zone of growth inhibition of the
test strains. As a result of bioscreening it was found
that the studied compound III showed a pronounced
Thus, proceeding from o- and p-hydroxyl-
benzhydrazides we obtained thiosemicarbazide deriva-
tives very promising in terms of biological activity,
among which are found substances with strong and
moderate antimicrobial activity.
EXPERIMENTAL
IR spectra were recorded on a Fourier-transform
AVATAR-320 NICOLET spectrometer from the
1
tablets with KBr. H NMR spectra were recorded on a
Table 2. Antimicrobial activity of samplesa
Compound
III
S. aureus 505
24±0.1
18±0.1
20±0.1
23±0.2
26±0.1
–
Bacillus Subtilis
26±0.1
20±0.1
20±0.2
22±0.1
24±0.1
–
Pseudomonas aeruginosa
E. coli M–17
20±0.1
14±0.1
11±0.2
17±0.1
23±0.2
–
Candida Albicans
14±0.2
18±0.2
11±0.3
–
–
IV
–
V
–
24±0.1
–
16±0.1
–
VII
Gentamicin
Nystatin
22±0.1
a
“–” means absence of the zone of growth inhibition. The diameters of zones of growth inhibition is less than 10 mm and the continuous
growth in the cup was assessed as the absence of antibacterial activity, 10–15 mm is weak activity, 15–20 mm is moderate activity, more
than 20 mm is expressed activity.
RUSSIAN JOURNAL OF GENERAL CHEMISTRY Vol. 82 No. 4 2012

SYNTHESIS AND ANTIMICROBIAL ACTIVITY
671
Bruker DRX500 spectrometer at a frequency of
500 MHz in a solution of DMSO-d6 relative to internal
TMS. Melting points were determined on a Boetius
unit. The reaction progress was monitored and the
purity of the compounds obtained was checked by
TLC on Silufol UV-254 plates in the system isopropyl
alcohol–benzene–25% ammonia solution 10:5:2. The
plates were developed with the iodine vapor.
and the mixture was stirred at room temperature for
about 2 h till the absence of glucosyl isothiocyanate
(TLC monitoring). The solution was evaporated to
give a white powdery substance. The yield of crude
substance 1.72 g (96%). After two recrystallizations
from benzene a crystalline product was obtained, mp
136–137°C.
4-(2,3,4,6-Tetra-O-acetyl-β-D-glucopyranosyl)-1-
(4-hydroxybenzoyl)thiosemicarbazide (VIII) was
obtained similarly to compound VII in 57% yield, mp
145–146°C (from 2-propanol).
4-Allyl-1-(2-hydroxybenzoyl]thiosemicarbazide
(III). To a stirred solution of 1.52 g (0.01 mol) of o-
hydroxybenzhydrazide in 20 ml of ethanol was added
dropwise 1.1 ml (0.011 mol) of allyl isothiocyonate.
The mixture was stirred for 60 min at 50–60°C. The
completion of the reaction was monitored by TLC. At
cooling a powdery white crystalline substance pre-
cipitated. The recrystallization from 2-propanol gave
2.14 g (85%) of compound III, mp 213–215°C.
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216°C (from 2-propanol).
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