Aromatic sulfonyl hydrazides and sulfonyl hydrazones: Antimicrobial activity and physical properties

Article abstract of DOI:10.1007/s00044-012-0104-0

A series of novel aromatic sulfonamide derivatives (1-7) were synthesized and characterized by elemental analyses, FT-IR, ¹H NMR, ¹³C NMR, and LC/MS techniques. The compounds' quantum mechanical descriptors, surface area, and molecul

Full text of DOI:10.1007/s00044-012-0104-0

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res (2013) 22:1330–1338
DOI 10.1007/s00044-012-0104-0
ORIGINAL RESEARCH
Aromatic sulfonyl hydrazides and sulfonyl hydrazones:
antimicrobial activity and physical properties
•
H. Gu¨zin Aslan Nurcan Karacan
Received: 29 September 2011 / Accepted: 22 May 2012 / Published online: 14 June 2012
Ó Springer Science+Business Media, LLC 2012
Abstract A series of novel aromatic sulfonamide deriv-
atives (1–7) were synthesized and characterized by ele-
mental analyses, FT-IR, 1H NMR, 13C NMR, and LC/
MS techniques. The compounds’ quantum mechanical
descriptors, surface area, and molecular volume were cal-
culated. All the synthesized compounds were evaluated in
vitro as antimicrobial agents against representative strains
of gram-positive and gram-negative bacteria and as anti-
fungal agents for yeast by both the disc diffusion and
minimal inhibition concentration methods for comparison.
All the bacteria and fungi studied were screened against
some commercial antibiotics to compare with our chemi-
cals’ zone diameters. Quantitative structure–activity rela-
tionship studies with multiple linear regression analysis
were applied to find the correlation between the different
calculated molecular descriptors of the synthesized com-
pounds and biological activity.
immunocompromised patients (HIV infection, antitumoral
treatments, organ transplant-associated immunosuppressive
therapy) as well as patients undergoing more invasive
medical procedures (extensive surgery, prosthetic implants),
among others (Georgopapadakou and Walsh, 1996).
A major associated problem is that the incidence of
drug-resistant isolated bacterial strains in the community
has become quite alarming. For these reasons, the demand
for new and better chemotherapic compounds has
increased, and nowadays the search for new chemicals with
antimicrobial activity has become an important field of
research (Chohan et al., 2006; Bruggraber et al., 2004;
Mills, 2006).
Sulfonamides, and their different derivatives are exten-
´
sively used in medicine (Borras et al., 2004) due to their
pharmacological antibacterial activity. They interfere with
the use of p-aminobenzoic acid (PABA) in the biosynthesis
of tetrahydrofolic acid, which is an essential growth factor
for the bacteria’s metabolism (Anand, 1996). In spite of the
fact that the emergence of drug-resistant strains is one of
the principal constraints of sulfonamide therapy, it has
continued unabated. For instance, sulfonamides are con-
sidered as the drugs of choice for the treatment of nocar-
diosis and are mainly used for the treatment of urinary tract
and methicillin-resistant bacteria infections (Anand,1996;
Foye et al., 1995).
Keywords Antimicrobial activity ꢀ Sulfonamides ꢀ
Aromatic sulfonamide ꢀ MIC
Introduction
During the past two decades, the frequency and types of
life-threatening infections has increased. In addition to the
usual kinds of cases, there have been increasing numbers of
Amsacrine (C21H19N3O3S) is used in cancer chemo-
therapy (Lomax and Narayanan, 1988; Jensen et al., 1990;
Finlay et al., 1990; Rutner et al., 1974). Some sul-
fonylhydrazines are known to have antineoplastic effects
which prevent malignant cells from growing and spreading
(Shyam et al., 1990).
H. G. Aslan (&)
Department of Chemistry, Science Faculty, Erciyes University,
38039 Melikgazi, Kayseri, Turkey
e-mail: guzina@erciyes.edu.tr
This prompted us to synthesize a series of novel sym-
metric aromatic sulfonamides. Therefore, we obtained a
series of new sulfonamides (1–7) and characterized their
N. Karacan
Department of Chemistry, Science and Arts Faculty,
Gazi University, 06500 Ankara, Turkey
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Med Chem Res (2013) 22:1330–1338
1331
structures by FT-IR, 1H NMR, 13C NMR, and LC/MS
techniques. We investigated their antibacterial activities
against gram-positive (Staphylococcus aureus ATCC
25923, Bacillus cereus ATCC 11778, Enterococcus
faecalis ATCC 29212, Bacillus subtilis ATCC 6633,
Staphylococcus epidermidis ATCC 12228, Enterobacter
aerogenes ATCC 13048) and gram-negative bacteria
(Pseudomonas fluorescens ATCC 49838, Klebsiella pneu-
monia ATCC 13883, Escherichia coli ATCC 25922,
Pseudomonas aeruginosa ATCC 27853) and as antifungal
agents against Candida albicans (ATCC 90028), by both
the disc diffusion and minimal inhibition concentration
(MIC) methods for comparison. All the bacteria and fungi
studied were screened against proper antibiotics to com-
pare them with our chemicals’ zone diameters.
Results and discussion
Figures 1 and 2 illustrate the general synthetic route used to
prepare our compounds. The exothermic nucleophilic sub-
stitution reaction of benzene sulfonyl chloride with various
amines was employed to form benzenesulfonicacid-1-
methylhydrazide (1) (Dauban and Dodd, 2000; Newcombe,
1955, Kloes, 1959; Hrubiec et al., 1986), N-(3-amino-
2-hydroxypropylbenzenesulfonamide (2) (Karacan et al.,
2011) and N-(2-hydroxyethylbenzene sulfonamide (3).
Compound 1, which was treated with various aldehydes
and ketones, was synthesized to form salicylaldehyde benzene-
sulfonylhydrazone (4), 2-hydroxy-1-acetophenone benzene-
sulfonylhydrazone (5), 2-hydroxy-1-naphtaldehydebenzene-
sulfonylhydrazone (6), and thiophene-2-carbaldehyde
Fig. 2 Preparation of aromatic sulfonyl hydrazones
benzenesulfonyl hydrazone (7) (Aslan et al., 2011; Aslan,
2008). Among the different solvents used, THF helped to
facilitate a better nucleophilic substitution reaction for the
series of compounds.
NMR spectra
The NMR spectra of compounds were recorded in DMSO-
1
d6, using TMS as an internal standard. The H NMR and
13C NMR data for half of the compounds are given in
1
Tables 1, 2, 3, and 4, respectively. The H NMR spectrum
of compound 4 showed that the OH proton had a broad
peak at 10.30 ppm, and the N=CH proton at 8.60 ppm.
Based on these data, the structure of the phenol-imine
compound of 4 was found in the DMSO solution. On the
other hand, the IR spectrophotometric observation of (C–
O) stretching vibration at 1,258 cm-1 and (C=N) stretching
vibration at 1,659 cm-1 convinced us to propose that the
structure of the phenol-imine compound was in the solid
phase (Aslan et al., 2011). The 1H NMR spectrum of
compound 6 showed that OH protons had a broad peak at
12.85 ppm, a singlet at 9.98 around the imine zone and a
doublet at 8.66 ppm. Based on these data, the structures of
the ketone-amine and phenol-imine compounds of 6 in
DMSO solution indicate the presence of tautomeric forms.
On the other hand, the IR spectrophotometric observation
of (C–O) stretching vibration at 1,265 cm-1 and (C=N)
stretching vibration at 1,636 cm-1 convinced us to propose
that the structure of the phenol-imine was in the solid phase
(Aslan, 2008). The 1H NMR spectrum of the 1, 4, 5, 6, and
7 compounds showed that –N–CH3 protons had peaks at
Fig. 1 Preparation of aromatic sulfonyl hydrazides
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Med Chem Res (2013) 22:1330–1338
Table 1 The 1H NMR data (ppm) of the sulfonyl hydrazide
compounds
Table 4 The 13C NMR data (ppm) of the sulfonyl hydrazone
compounds
Assignment
Bsmh
Bsea
Bsdiap
Assignment Hsalbsmh
Hafbsmh
Covered
Hnafbsmh T2kbsmh
N–NH2
N–CH3
Ar H
4.85 (s)
–
–
N–CH3
Ar C
35.22
37
37
2.6 (s)
–
–
158.22–117.13 130.45–119.81 164–112
15.55
122–144
6.96–8.89 (s)
7.26–8.11
–
7.15 (w, 8H)
7.15 (w, 8H)
C–CH3
N=CH
NH–CH2
NH–CH2
–
–
141.61
133.60
161
156
2.83 (t, 2H,
8Hz)
2.88 (d, 2H,
3.36 Hz)
Table 5 Wave number (cm-1
compounds
) of selected vibration of the
NH–CH2–CH2
–
3.59 (t, 2H,
7.36 Hz)
–
CH2–NH2
–
–
–
7.15 (w, 8H)
3.87 (s, 1H)
Comp.
mNHas
mNHs
m(SO2)as
m(SO2)s
mC=N
NH–CH2–CH
CHOH–CH2
Bsmh
3,141
3,100 1,249 (w) 1,157 (m)
– 1,361 (w) 1,151 (m)
–
–
–
2.66 (d, 2H,
3.54 Hz.)
Bsea
3,290 (sh)
OH
–
5.18 (s, H)
5.28 (s, br)
Bsdiap
3,331 (sh) 3,360 1,334 (w) 1,156 (m)
Hsalbsmh
Hafbsmh
Hnafbsmh
T2kbsmh
–
–
–
–
–
–
–
–
1,295 (w) 1,180 (m) 1,612
1,329
1,344
1,316
1,150
1,162
1,166
1,606
1,636
1,617
Table 2 The 13C NMR data (ppm) of the sulfonyl hydrazide
compounds
m Stretching vibration, sh sharp, m medium, w weak
Assignment
Bsmh
Bsea
Bsdiap
N–CH3
40.46
FT-IR spectra
Ar C C1
125.95–148.13
125.77–134.0
45.42
129.07–128.04
42.47
NH–CH2
The selected vibration wave numbers of the IR spectra
compounds are listed in Table 5. The presence of NH2
vibrations in the prepared compounds 1 and 3, which are
observed between 3,100 and 3,250 cm-1 as double bonds
and the presence of a single and strong NH band observed
at *3,275–3,293 cm-1 confirms the presence of secondary
amine groups in our compounds (2–3). mas(SO2) and
ms(SO2) vibrations are observed between 1,249–1,361 and
NH–CH2–CH2
NH–CH2–CH
CHOH–CH2–
60.08
64.87
40.59
Table 3 The 1H NMR data (ppm) of the sulfonyl hydrazone
compounds
1,150–1,180 cm-1
.
Assignment Hsalbsmh Hafbsmh
Hnafbsmh
2.60
T2kbsmh
2.52
LC/MS spectra
N–CH3
2.94
2.69 (d, 3H,
8 Hz)
LC/MS shows that compounds 1 ([M]?), 2 ([M?H?]), 3
([M-H?]), 4 ([M?Na]?), 5 ([M-H]?), 6 ([M?H]?), and
7 ([M-H]?) give ions at 186.1, 231.2, 200.3, 313.1,
303.25, 341.2, and 279.80 m/z, respectively.
Ar H
6.85–7.65 7.67–6.94
8.05–7.31
7.18–8.75
8.84
C–CH3
2.69 (d, 3H,
6 Hz)
N=CH
OH
8.70
9.98 (s, H)
All compounds of the C6H5 group fragment separately
and this was observed at 79.2 m/z.
8.66 (d, H,
3.5 Hz)
10.25
12.93 (s, H)
12.85
Biological activity
In this study, the antimicrobial activities of the synthesized
compounds were screened against various pathogens in
vitro by the disc diffusion and microdilution methods.
Tetracycline, trimethoprim, miconazole, and nystatin were
used as positive controls against both bacteria and yeast
using the disc diffusion method. The results of the anti-
bacterial and antifungal studies of all the synthesized
2.60, 2.94, 2.69, 2.60, and 2.52 ppm, respectively. It is
known that rotation around a single bond brings together
the mixture of conformers in the solution.
NMR chemical shift calculations for the most stable
conformers were performed using the GIAO approach in
order to correctly assign proton and carbon peaks, which
are now widely used in efficient assignment.
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Table 6 The MICs of the tested compounds and reference antibiotics
MIC (lg/mL)
Tested compound number:
1
2
3
4
5
6
7
8
9
10
11
S. aureus ATCC 25923
B. cereus ATCC 11778
E. faecalis ATCC 29212
B. subtilis ATCC 6633
125
125
[2,000
[2,000
[2,000
[2,000
[2,000
1,000
[2,000
[2,000
[2,000
1,000
250
500
500
500
250
500
500
500
250
500
250
500
500
250
250
500
250
500
250
250
0.000
0.000
0.625
0.075
0.125
0.150
0.125
0.150
0.075
0.125
0.000
1.000
0.000
0.500
1.000
0.000
1.000
1.000
1.000
0.000
1.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.150
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.000
0.125
250
1,000
500
1,000
125
125
S. epidermidis ATCC 12228
E. aerogenes ATCC 13048
P. fluorescens ATCC 49838
K. pneumonia ATCC 13883
E. coli ATCC 25922
125
[2,000
1,000
500
125
125
1,000
500
1,000
1,000
1,000
1,000
1,000
500
250
62.5
125
1,000
1,000
125
[2,000
1,000
1,000
1,000
500
125
125
1,000
125
P. aeruginosa ATCC 27853
C. albicans ATCC 90028
62.5
[2,000
1,000
1,000
1,000
500
125
1,000
1,000
1,000
8 Tetracycline (lg/mL), 9 trimethoprim(lg/mL), 10 miconazole (lg/mL), 11 nystatin (lg/mL)
compounds are listed in Tables 6 and 7. As seen in
Table 6, compound 1 is the most potent sulfonyl hydrazide
of this series. It showed good antibacterial activity against
both gram-negative and gram-positive bacteria, and has
also shown antifungal activity against C. albicans at
[2,000 lg/mL. Similarly, sulfonylhydrazone (7) showed
observable activity against gram-negative bacteria and
gram-positive bacteria. As shown in Table 6, the sulfonyl
hydrazones which are part of the –OH group show anti-
fungal activity against C. albicans. Using the disc diffusion
method we found that compound 6 displays good activity
against all tested microorganisms except E. coli.
Table 7 Inhibition zones (diameter) in mm of compound and refer-
ence antibiotic discs against tested microorganisms by the disc dif-
fusion method
˙
Bacteria and fungi
Inhibition zone (mm, 140 lg/disk)
1
2 3 4
5
6
7
8
9
10 11
S. aureus ATCC 25923
B. cereus ATCC 11778
19 0 0 13 10 19 12 0 37
0
0
0
0
0
0
0
0
0
0
15 0 0 11 10 26 25 19
0
E. faecalis ATCC 29212 15 0 0 11 9 17
0
0 27
B. subtilis ATCC 6633
16 0 0
9
9 28 24 26 20
S. epidermidis ATCC
12228
14 0 0 14 12 16 15 19 29
E. aerogenes ATCC
13048
16 0 0
22 0 0 12 11 26 25 19
17 0 0 6 23 21 12
6
5 18 17 0 28
0
0
0
0
0
0
Different quantum mechanical descriptors including
dipole moment, energies of the frontier orbitals (HOMO,
LUMO), refractivity, and dipole moment were calculated
by HyperChem 7.5 software.
P. fluorescens ATCC
49838
0
0
0
K. pneumonia ATCC
13883
6
The log P value definition is an important method used
to determine antimicrobial activity. The octanol–water
partition coefficient (log P value) of a drug substance is an
indicator of compound lipophilicity and solubility. A
higher log P value increases the lipophilic property of
compounds. According to the Lipinski (Lipinski et al.,
2001) and Ghose (Ghose et al.,1999; Brendan et al., 2007)
rules, the compound’s log P value must be between -0.4
and ?5.6. In our study, the activity in the sulfonamides (1–
3) increased with increasing log P value. Similarly, the
antimicrobial activity in the sulfonyl hydrazones (5–6)
increased with increasing log P value. However, com-
pound 7 showed the highest activity and lowest log P value
because this compound was located in the thiophene ring,
instead of in the phenol group.
E. coli ATCC 25922
18 0 0 13 11 17 17 23
19 0 0
0
0
0
0
P. aeruginosa ATCC
27853
6
5 25 24 23 31
C. albicans ATCC 90028 0 0 0 14 12 16
0
0
0 26 18
8 Tetracycline (lg/mL), 9 trimethoprim(lg/mL), 10 miconazole
(lg/mL), 11 nystatin (lg/mL)
properties of molecules. In Figs. 3 and 4, the blue areas on
the maps represent the positive potential values, and the red
regions correspond to the negative potential values.
In this study, QSAR analysis was performed by the
stepwise multiple linear regression method. The statiscially
significant QSAR models are given below.
QSAR model for antimicrobial activity against Candida
albicans
Electronic potential maps enable us to visualize the
charge distributions of molecules; charge is related to the
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Med Chem Res (2013) 22:1330–1338
Fig. 3 Frontier molecular
orbitals of sulfonyl hydrazides
and sulfonyl hydrazones (Color
figure online)
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Med Chem Res (2013) 22:1330–1338
1335
Benzenesulfonicacid-1-
methylhydrazide
N-(3-amino-2-
hydroxypropylbenzenesulfonamide sulfonamide
N-(2-hydroxyethylbenzene
Salicylaldehyde
benzenesulfonylhydrazone
2-Hydroxy-1-acetophenon
benzenesulfonylhydrazone
2-Hydroxy-1-naphtaldehyde
benzenesulfonylhydrazone
Thiophene-2-carbaldehyde
benzenesulfonyl hydrazone
Fig. 4 Optimized geometries of aromatic sulfonyl hydrazides and aromatic sulfonyl hydrazones (Color figure online)
spectra were recorded on a Mattson-1000 FT-IR spectro-
ꢁ log MIC ¼ fꢁ2:676ðꢂ0:08Þg þ f V9 3:586ðꢂ0:639Þ
ꢁ V10 0:150ðꢂ0:007Þ g
photometer with samples prepared as KBr pellets. The
NMR spectra were obtained on a Bruker-Spectrospin
Avance DPX-400 Ultra-Shield (400 MHz) using DMSO-d6
and CDCl3 as solvents and TMS as an internal standard.
LC/MS-APC1 was acquired with an AGILENT 1100.
Melting point, were recorded on an Opti MELT 3 hot stage
apparatus. TLC was conducted on 0.25 mm silica gel
plates (60F254, Merck). The synthesized compounds
(except 4) were purified by column chromatography.
All calculations reported here in were carried out with
the GAUSSIAN 03 software (Frisch et al., 2003). Quantum
chemical descriptors (HOMO, LUMO, dipole moment,
heat of formation) were calculated by using the B3LYP/6-
31** method with the same software. The surface areas
(approx) and molecular volumes of the compounds were
calculated by using the HyperChem Release 7.5 package
program (HyperChem 7.5 program, 2002).
n ¼ 7; r2 ¼ 0:955; s ¼ 0:002; F ¼ 42:548;
V9 ¼ HOMO, and V10 ¼ LUMO SPRESS ¼ 0:307:
HOMO and LUMO are responsible for the formation of
many charge transfer complexes. According to the frontier
molecular orbital theory (FMO) of chemical reactivity, the
formation of a transition state is due to an interaction
between the frontier orbitals (HOMO and LUMO) of the
reacting species. Thus, the treatment of the frontier
molecular orbitals separately from the other orbitals is
based on the general principles governing the nature of
chemical reactions (Fukui, 1975).
The statistical analyses were performed with SPSS
software (version 13, 2002). The abbreviations for the
statistical parameters given for each equation were:
n (number of data points), r2 (squared correlation coeffi-
cient), and s (standard error of estimate), SPRESS (sum of
square standard error), and F (Fischer statistics).
General procedure for the synthesis
Statistically, a significant relationship was observed
between the antimicrobial activity of sulfonyl hydrazides
and sulfonyl hydrazones against Candida albicans (ATCC
90028) and HOMO–LUMO.
The nucleophilic substitution reaction of the different ali-
phatic diamines with benzene sulfonyl chloride was carried
out as follows (only compounds 1–3): A THF solution of
aromatic sulfonyl chlorides (Ar-SO2Cl) was added drop-
wise to the THF solution of methyl hydrazide (1:2 equiv),
maintaining the temperature between -5 and -10 °C.
Then, the reaction mixture was stirred for 24 h at room
temperature (completion of the reaction was monitored by
TLC). After the completion of the reaction, the solvent was
Experimental methods
Elemental analyses (C, H, N, and S) were performed on a
LECO-CHSNO-9320 type elemental analyzer. The IR
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Med Chem Res (2013) 22:1330–1338
removed under vacuum and the solid residue was purified
by column chromatography. Compounds (4–7) were syn-
thesized according to the following general procedure: A
THF solution of various aldehydes and ketones was added
to the THF solution of compound 1 maintaining the tem-
perature at -5 and -10 °C. Then the reaction mixture was
stirred for 48 h at room temperature (completion of the
reaction was monitored by TLC). After completion of the
reaction, the solvent was removed under vacuum. The solid
residue was purified by column chromatography (except
for compound 4).
C, 57.931; H, 4.880; N, 9.663; S, 11.022. Found: C,
57.916; H, 4.860; N, 9.648; S, 11.042.
Synthesis of 2-hydroxy-1-acetophenone
benzenesulfonylhydrazone (5)
The general synthetic method described above afforded a
yellow solid from bsmh (3.95 mL, 47.5 mmol) and
1-hydroxy acetophenone (3.0 mL, 23.7 mmol). The product
was purified by column chromatography. The results were as
follows: yield 68 %; mp 194–195 °C; analytical calculation
for C15H16N2SO3: C, 59.201; H, 5.289; N, 9.212; S, 10.525.
Found: C, 59.193; H, 5.298; N, 9.203; S, 10.533.
Synthesis of benzenesulfonicacid-1-methylhydrazide (1)
The general synthetic method described above afforded a
white solid from methylhydrazide (4.4 mL, 0.08 mol) and
benzenesulfonyl chloride (5.05 mL, 0.04 mol). The prod-
uct was purified by column chromatography. The results
were as follows: yield 62 %; mp 67–68 °C; analytical
calculation for C7H10N2O2S: C, 45.090; H, 5.430; N,
15.090; S, 17.190. Found: C, 45.140; H, 5.410; N, 15.040;
S, 17.210.
Synthesis of 2-hydroxy-1-naphtaldehyde
benzenesulfonylhydrazone (6)
The general synthetic method described above afforded a
yellow solid from bsmh (4.7 mL, 47.4 mmol) and
2-hydroxy-1-naphthaldehyde (3.0 mL, 23.7 mmol). The
product was purified by column chromatography. The results
were as follows: yield 70 %; mp 259–262 °C; analytical
calculation for C18H16N2SO3: C, 63.523; H, 4.732; N, 8.234;
S, 9.408. Found: C, 63.512; H, 4.737; N, 8.229; S, 9.419.
Synthesis of N-(3-amino-2-
hydroxypropylbenzenesulfonamide (2)
Synthesis of thiophene-2-carbaldehyde benzenesulfonyl
hydrazone (7)
The general synthetic method described above afforded a
white solid from benzenesulfonyl chloride (5.05 mL,
0.04 mol) and 1.3 diaminopropan-2-ol (7.21 g, 0.08 mol).
The product was purified by column chromatography. The
results were as follows: yield 51 %; mp 155 °C; analytical
calculation for C9H14N2O3S: C, 47.000; H, 6.130; N,
12.104; S, 13.910. Found: C, 46.940; H, 6.127; N, 12.164;
S, 13.922.
The general synthetic method described above afforded a
yellow solid from bsmh (2.0 mL, 16.0 mmol) and thionyl
chloride (1.06 mL, 8.1 mmol). The product was purified by
column chromatography. The results were as follows:
yield 35 %; mp 83.7 °C; analytical calculation for
C12H12N2S2O2: C, 41.428; H, 4.326; N, 9.979; S, 22.851.
Found: C, 41.409; H, 4.314; N, 9.991; S, 22.870.
Synthesis of N-(2-hydroxyethylbenzene sulfonamide (3)
Biological activity: in vitro evaluation
The general synthetic method described above afforded a
white solid from benzenesulfonyl chloride (5.05 mL,
0.04 mmol) and ethanolamine (4.83 mL, 0.08 mol). The
product was purified by column chromatography. The
results were as follows: yield 65 %; mp 79–80 °C; ana-
lytical calculation for C8H11NO2S: C, 47.700; H, 5.610; N,
7.000; S, 15.833. Found: C, 47.740; H, 5.519; N, 6.960; S,
15.931.
The biological activity of the synthesized sulfonamide
derivatives was individually screened against a panel of
microorganisms, including gram-positive (Staphylococcus
aureus ATCC 25923, Bacillus cereus ATCC 11778,
Enterococcus faecalis ATCC 29212, Bacillus subtilis ATCC
6633, Staphylococcus epidermidis ATCC 12228, Entero-
bacter aerogenes ATCC 13048), and gram-negative bacteria
(Pseudomonas fluorescens ATCC 49838, Klebsiella pneu-
monia ATCC 13883, Escherichia coli ATCC 25922, Pseu-
domonas aeruginosa ATCC 27853) and yeast (Candida
albicans ATCC 90028). The cultures were obtained from
Erciyes University’s Department of Biology. Bacterial
strains were cultured overnight at 35 °C in Triptic Soy Broth
(TSB), and the yeast was cultured overnight at 30 °C in
Yeast Peptone Dextrose Broth (YPDB) in a rotary shaker at
Synthesis of salicylaldehyde benzenesulfonylhydrazone (4)
The general synthetic method described above afforded a
yellow solid from bsmh (3.1 mL, 46.0 mmol) and salicyl-
aldehyde (3 mL, 23.0 mmol). The product was crystallized
from acetonitrile. The results were as follows: yield 59 %;
mp 167–168 °C; analytical calculation for C14H14N2SO3:
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Med Chem Res (2013) 22:1330–1338
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200 rpm. Overnight cultures were then transferred to fresh
media and the turbidity of all broth cultures was adjusted to
0.1 absorbance at 640 nm on a spectrophotometer. These
stock cultures were stored in the dark at 4 °C during the
survey.
adjusted to the OD640 0.1 (equivalent to 0.5 Mc Farland
Turbidity Tubes) was swabbed over the entire surface of the
medium. Sterile 6-mm-diameter disks (1.2 mm thickness
Whatman filter paper) were impregnated with 50 lL dis-
solved extracts (including 1.2 mg total extract per disk) and
placed onto MHA or YPDA. Negative controls were pre-
pared by just using DMSO. For the bacterial cells, ampicillin
(50 lg/disk), trimethoprim (30 lg/disk), and tetracycline
(30 lg/disk) on MHA and for the yeast miconazole (40 lg/
disk) and nystatin (30 lg/disk) on YPDB were used as ref-
erence standards to determine the sensitivity of each strain.
The inoculated plates were incubated at 35 °C for 18–24 h.
The antibacterial activity was measured as the diameter
(mm) of the clear zone of growth inhibition. Four disks per
plate were used and each test was run in duplicate.
Minimal inhibitory concentration
MIC were determined by the micro dilution broth method
following the procedures recommended by the National
Committee for Clinical Laboratory Standards (NCCLS,
2000). Briefly, synthesized sulfonamide derivatives dis-
solved in dimethylsulfoxide (DMSO) were first diluted to
the highest concentration, and then serial twofold dilutions
were made in a concentration range from 31.25 to
2,000 lg/mL in a sterile 2.4 mL 96-well Masterblock dish
containing either MHB for the bacterial strains or YPDB
for the yeast. For the test, the 96-well plates were prepared
by dispensing into each well 195 lL of the serial dilutions
of the synthesized sulfonamide derivatives which were
then transferred into seven consecutive wells in each col-
umn. The last well, containing only 195 lL of MHB or
YPDB without compound, was used as a negative control.
For each column of the plate, 5 lL of the strains to be
tested was inoculated into each well individually. The final
volume in each well was brought up to 200 lL. For the
bacterial cells, ampicillin, trimethoprim, and tetracycline in
MHB and for the yeast miconazole and nystatin in YPDB
at the concentration range of 1,000–7.8 lg/mL were used
as standard drugs for positive control. The contents of each
well were mixed on a plate shaker at 300 rpm for 20 s and
then incubated at 35 °C for 18–24 h. Microbial growth was
determined by measuring absorbance at 600 nm using the
Tecan Sunrise microplate reader and with the presence of a
white ‘‘pellet’’ on the well bottom for visual examination.
Concentrations resulting in no growth were confirmed by
plating 5 lL samples from the clear wells on MHA or
YPDA. The compounds tested in this work were screened
twice against each organism. The MIC was defined as the
lowest concentrations of the antimicrobial agents that
inhibited visible growth of the microorganism.
Conclusion
In conclusion, a series of aromatic sulfonyl hydrazides and
sulfonyl hydrazones (1–7) were synthesized, and their
antimicrobial activities were evaluated. All compounds
demonstrated potent inhibition against all the strains tested.
Further research in this area is in progress in our laboratory.
Consequently, these compounds may be recommended for
industrial application.
Acknowledgments The authors are grateful to the TUBITAK
Research Foundation for financial support under project number 104
T 390.
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