Synthesis and evaluation of in vitro antimicrobial activity of novel 2,3-disubstituted-4-thiazolidinones from fatty acid hydrazides

Article abstract of DOI:10.1007/s00044-012-0326-1

The efficient cyclocondensation reaction of fatty acid hydrazides with thioglycolic acid (TGA)/mercapto acetic acid and Thio lactic acid under anhydrous condition in the presence of catalyst anhydrous ZnCl₂ in dimethyl formamide solvent is desc

Full text of DOI:10.1007/s00044-012-0326-1

MEDICINAL
CHEMISTRY
RESEARCH
Med Chem Res
DOI 10.1007/s00044-012-0326-1
ORIGINAL RESEARCH
Synthesis and evaluation of in vitro antimicrobial activity of novel
2,3-disubstituted-4-thiazolidinones from fatty acid hydrazides
•
Himani Varshney Aiman Ahmad Nida N. Farshori
•
•
•
Anis Ahmad Asad U. Khan Abdul Rauf
•
Received: 17 July 2012 / Accepted: 6 November 2012
Ó Springer Science+Business Media New York 2012
Abstract The efficient cyclocondensation reaction of
fatty acid hydrazides with thioglycolic acid (TGA)/mer-
capto acetic acid and Thio lactic acid under anhydrous
condition in the presence of catalyst anhydrous ZnCl2 in
dimethyl formamide solvent is described. The synthesis
provides the product, substituted thiazolidinone, in appro-
priate yield. The synthesized compounds were character-
ized by IR, 1H NMR, 13C NMR, and mass spectra and these
compounds were tested for in vitro antibacterial activity
against the representative group of Gram-positive and
Gram-negative bacteria such as Pseudomonas aeruginosa
(ATCC-27853), Escherichia coli (ATCC-25922), Strepto-
coccus pyogenes, and Klebsiella pneumonia (Clinical iso-
late), Methicillin resistant Staphylococcus aureus (MRSA
?ve), and also tested for antifungal activity against
Aspergillus fumigates, Trichophyton mentagrophytes, Pen-
icillium marneffei, Candida albicans. Minimum inhibitory
concentration, minimum bacterial concentration, and min-
imum fungicidal concentration were determined for the test
compounds along with the reference standards used.
Abbreviations
TGA
TLA
TLC
DMF
Thioglycolic acid
Thio lactic acid
Thin layer chromatography
Dimethyl formamide
DMSO Dimethyl sulfoxide
Introduction
Thiazolidinone derivatives have vast medicinal importance
in the field of pharmaceutical chemistry (Verma and Saraf,
2008). 4-Thiazolidinone, a saturated form of thiazole with
carbonyl group on fourth carbon, has been assumed as a
magic moiety (wonder nucleus) with all types of pharma-
cological activities such as anti-HIVagent (Pareek et al.,
2011), antidiarrheal, anticonvulsant, antidiabetic, antihista-
minic, anticancer, Ca?2 channel blocker, PAF antagonist,
antitubercular (Dubey et al., 2010), antioxidant, hypo-
glycemic, inhibition of gastric H? K?-ATPase, CFTR
(cystic fibrosis transmembrane conductance regulator pro-
tein) inhibitor (Ma et al., 2002), antiplatelet-activating factor
a-antagonist, antifungal (Mistry and Desai, 2004), and
antituberculosis activity. 4-Thiazolidinone derivatives also
have the good antimicrobial activity. Morbidity and mor-
tality are the most common enteric bacterial infections
mostly in Indian subcontinent areas, tropical part of Africa
(Qadri et al., 2005) and part of South America (Devasia et al.,
2006). The world’s most common and serious infectious
diseases like dysentery and diarrhea (Zhang et al., 2006;
Shaheen et al., 2004) are caused by Escherichia coli,
symptoms of these diseases are stomachache, cramps,
bloating, or tenderness (Yoder et al., 2006). In spite of these
Keywords Substituted thiazolidinone ꢀ
Thioglycolic acid ꢀ Cyclocondensation reaction ꢀ
Antimicrobial activity
H. Varshney ꢀ A. Ahmad ꢀ N. N. Farshori ꢀ A. Rauf (&)
Department of Chemistry, Aligarh Muslim University,
Aligarh 202 002, India
e-mail: abduloafchem@gmail.com
A. Ahmad ꢀ A. U. Khan
Interdisciplinary Biotechnology Unit, Aligarh Muslim
University, Aligarh 202 002, India
123

Med Chem Res
diseases, abscess of the brain is a major problem of E. coli
infection (Sonntag et al., 2004). Amoxicillin, norfloxacin,
and ciprofloxacin are generally used in case of E. coli
infections, but have harsh side effects (Puerto et al., 2006).
The main causes of treatment failure (Nolan et al., 1979) are
the toxicity and resistance to the drugs. Human’s mucosal
health is affected by the different parasitic bacteria such as
S. aureus, S. pyogenes, S. typhimurium, E. coli. The Infection
with these microorganisms may have significant impact on
huge demolition of host tissues and severe diseases (Chee-
seman et al., 1979, Porter et al., 1984). Above 90 % of the
cases of vaginitis are of candidiasis, trichomoniasis, and
bacterial vaginosis (Cruickshank et al., 1975).
ZnCl2 provided the product in minimum reaction time and
appropriate yield. The synthesized compounds were char-
1
acterized by IR, H NMR, 13C NMR, and MASS spectral
data. In the IR spectrum of the compound, the disappearance
of peak of the free NH2 group supports the formation of
intermediate product by the condensation of fatty acid
hydrazides with aromatic aldehydes. The purity of the
compound was monitored by TLC and the synthesized
compounds were screened for antimicrobial activity, and
found to be the potent antibacterial and antifungal agent. The
synthetic pathway for the synthesis of substituted 4-thiazo-
lidinone is presented in scheme 1. A detailed spectral
description for compound 2b and 3b is discussed below.
IR spectrum of compound 2b shows a characteristic
peak at 3,220 cm-1(N–H stretching), 1,664 cm-1 (corre-
The thiazolidinone derivatives have been found to be the
inhibitor of HCV-NS5B (Hepatitis C virus-non structural
protein) polymerase (Rawal et al., 2008), reported by
Kaushik-Basu et al., therefore the thiazolidinone deriva-
tives were now used as potent anti-NS5B agent (Kaushik
Basu et al., 2008).
1
sponding to carbonyl C–O stretching). H NMR was more
informative in assigning the structure. Diagnostic peaks at
d 5.42 and 2.54 were observed for the thiazolidinone ring
system. A multiplet at d 7.68 was observed for the aromatic
ring. A singlet at d 8.80 was observed for NH proton. The
methine proton of C-9 showed a signal at 5.79. The C-10
methylene protons designated as HE and HZ displayed two
distinct d values when coupled with adjacent C-9 methine
Because of the biological importance of thiazolidinone
derivatives as calcium agonists and HIV-RT inhibitors, the
solid phase and solution synthesis of 4-thiazolidinone have
been widely described usually in an one-pot reaction
involving an amine, an aldehyde, and a a-mercapto acid
(Colombo et al., 2008). An alternative to the use of
a-mercapto acids is via a Mannich reaction with formalde-
hyde and an amine. The above-mentioned cyclocondensa-
tion can be either run in one pot or in two steps with
prolonged heating in toluene or benzene (Pratap et al., 2011).
This cyclocondensation reaction can be accelerated by the
use of catalysts like N,N0-dicyclohexyl carbodiimide (DCC)
(Srivastava et al., 2002), O-(benzotriazol-yl)-N,N,N0,
N0-tetramethyl uronium hexafluoro phosphate (HBTU)
(Rawal et al. 2008), ferrite (Sadashivaa et al., 2009), zinc
chloride (Srivastava and Srivastava, 2000), sodium sulfate
(Sharma and Kumar, 2000), [bmim][PF6] (Yadav et al.,
2009; Zhang et al., 2009), and activated fly ash (Kanaga-
rajana et al., 2009). On the basis of above-mentioned facts, in
this paper, we report the synthesis of 2,3-disubstituted-4-
thiazolidinone derivatives of fatty acid hydrazides as good
antibacterial and antifungal agents.
1
protons. Thus, the H NMR showed two doublets of dou-
blet at d 5.01 and 4.94 for HZ and HE protons respectively.
In the 13C NMR peaks at d 169.84, 139.18 and 123.3 were
observed for the ring carbon atoms. The mass spectrum
was also consistent with the assigned molecular formula.
IR spectrum of compound 3b shows a characteristic
peak at 3,222 cm-1 (N–H stretching) and 1,641 cm-1
1
(carbonyl C–O stretching). H NMR spectra of the com-
pound show the following characteristic peaks, a singlet at
d 8.79 for NH proton and doublet at d 1.93 for CH3 proton.
13C NMR spectra also show characterization peaks at d
10.98 for methyl carbon along with the peaks at d 170.5,
138.1, and 129.4 for ring carbons.
Pharmacology
In literature, 4-thiazolidinone derivatives were found to be
the good antimicrobial agents (Mahyavanshi and Marjadi,
2011), therefore the synthesized long chain hydroxyl and
non-hydroxyl fatty acid derivatives of 4-thiazolidinone
were tested against the bacterial and fungal strains.
Result and discussion
Antifungal and antibacterial activity of the synthesized
compounds was performed by the disk diffusion method and
measured by Halo Zone Test. The minimum inhibitory
concentration (MIC) of synthesized compounds against
bacterial and fungal strains was performed by micro dilution
test. The results of these tests were observed visually and
spectrophotometrically. The investigation of antibacterial
screening data revealed that all the Screened compounds
Chemistry
In this paper, we report the synthesis of substituted 4-thia-
zolidinones, 2a–e and 3b by the reaction of mercapto acetic
acid and thiolactic acid with various saturated and olefinic
(internal and terminal) fatty acid hydrazides (1a–e), under
anhydrous condition in the presence of catalytic amount of
anhydrous ZnCl2 in DMF. The use of catalytic amount of
123

Med Chem Res
+
CHO
Cl
NH2
R
1a-e
Dry benzene
Reflux
Cl
CH
N
R
Anhydrous
Anhydrous
ZnCl2,
ZnCl2,
DMF,
DMF ,
Thiolactic acid
Thioglycolic acid
Cl
N
C
CH
S
R
CH
N
R
Cl
S
C
O
O
CH3
3b
2a-e
COMPOUND
R
O
NH
11
1a, 2a
HZ
HE
NH
3
O
1a, 2b, 3b
O
NH
5
5
1c, 2c
OH
O
NH
5
3
1d, 2d
1e, 2e
NH
5
2
O
OH
Scheme 1 Synthesis of 2,3-disubstituted-4-thiazolidinones 2a-e and 3b
showed moderate to good antibacterial activity. All the
synthesized compounds showed good inhibition against
E. coli, S. aureus, S. pyogenes species. Compound 2a, 2b,
and 2c showed good antibacterial activity nearly equivalent
to that of chloramphenicol.
The antifungal screening data showed only moderate
activity. Among the screened compounds, compounds 2a,
2b, and 2c showed excellent antifungal activity, nearly
equivalent to the control compound against C. albicans,
A. fumigatus, and P. marneffei fungal strains. The MFC of
123

Med Chem Res
few compounds was found to be the same as MIC and in
few compounds it was two- to fourfolds higher than the
corresponding MIC results. Compounds 2a, 2b, and 2c
exhibited stronger fungicidal activity against A. fumigates,
P. marneffei, and Candida albicans fungal strains.
presence of catalytic amount of anhydrous ZnCl2. After
completion, the reaction mixture was then cooled and
filtered. The solid was removed and the filtrate was
poured into crushed ice, filtered, and the crude product
was recrystallized with ethanol. The characterization data
of compounds 2a–e is given below.
Experimental
3-(Amido hexadec)-2-(40-chlorophenyl)-
4-thiazolidinone (2a)
Chemicals and instruments
White powder; Yield: 85 %; m.p. 129–130 °C; IR (KBr):
3217 (N–H amide), 2919 (C–H asymm.), 2851 (C–H
symm.), 1664 (C=O amide), 1600 (N–H bending), 1173
Undec-10-enoic acid (purity 98 %) and cis-9-octadecanoic
acids (purity 97 %) were purchased commercially from
Fluka Chemicals, Switzerland. (9Z, 12R)-12-Hydroxy
octadec-9-enoic (Ricinolic) and (9R, 12Z)-9-Hydroxy
octadec-12-enoic (Isoricinolic) acids were isolated from
seeds, laboratory grade (LR) chemicals were used for
extraction of oil from naturally occurring seeds. For the
synthesis, anhydrous conditions were required which were
achieved by drying the flask and other equipments in the
oven. The reactions were monitored by TLC (glass plates
coated with silica gel 60–80 mesh for thin layer chroma-
tography). The purity of the compounds was also determined
by thin layer chromatography (TLC) glass plates (20 9
5 cm) coated with a layer of silica gel-G (Merck, Mumbai,
India, 0.5-mm thickness) in the solvent system petroleum
ether : diethyl ether : acetic acid (50:50:1 v/v). The spots of
the compounds were observed on exposure to iodine vapor.
The melting points of the synthesized compounds were
determined by the open tube capillary method and are
uncorrected. The IR spectrums of the synthesized compound
were obtained on FT-IR spectrometer (KBr-pellets). The 1H
and 13C NMR spectra of compounds were recorded on a
Bruker Advance II 400 spectrometer and 1H NMR spectrum
of the compounds were recorded at 400 MHz and 13C NMR
spectrum were recorded at 100 MHz, Chemical shifts (d) are
quoted in ppm.
1
(C–C aliphatic), 639 (C–Cl aromatic ring substition); H
NMR (400 MHZ, CDCl3, d, ppm): 8.81 (s, 1H, NH), 7.83
(d, 2H, J = 8.80 HZ, Ar 2H), 7.37 (d, 2H, J = 8.52 HZ,
Ar 2H), 5.37 (s, 1H, NCHS), 2.74 (unresolved dd cen-
tered at, 2H, COCH2S), 2.52 (t, 2H, J = 4.14 HZ,
CH2CO), 1.70 (m, 2H, CH2CH2CO), 1.37 (br.s, 22H,
(CH2)11), 0.87 (dt, 3H, CH2CH3); 13C NMR (100 MHZ,
CDCl3, d, ppm): 169.6, 139.2, 129.3, 114.1, 77.3, 77.2,
77.0, 76.8, 34.1, 32.5, 30.0, 29.2, 28.9, 25.3, 23.1, 13.9.
MS (ESI): m/z = 489.120 [M ? Na]?, Calculated =
490.065.
3-(Amido undec-10-enyl)-2-[40-chlorophenyl]-4-
thiazolidinone (2b)
White powder; Yield: 74 %; m.p. 99–100 °C; IR (KBr):
3220 (N–H amide), 2923 (C–H asymm.), 2852 (C–H
symm.), 1664 (C=O amide), 1601 (N–H bending), 1540
(C=C aromatic), 1185 (C–C aliphatic), 666 (C–Cl aro-
1
matic ring substitution); H NMR (400 MHZ, CDCl3, d,
ppm): 8.80 (s, 1H, NH), 7.83 (d, 2H, J = 8.40 HZ, Ar
2H), 7.52 (d, 2H, J = 8.48 HZ, Ar 2H), 5.79 (tdd, 1H,
JHꢁ CH
9
= 6.70 HZ, J
CH2 = CH), 5.42 (s, 1H, NCHS), 5.01 (dd, 1H, J
= 10.10 HZ, JHꢁH = 16.90 HZ,
H–Hz
E
2
=
Hz–H
10.10 HZ, JH ꢁH = 2.80 HZ, HC=CH), 4.94 (dd, 1H,
z
E
Z
Chemistry: synthesis of fatty acid hydrazides (1a–e)
J
H ꢁH= 16.90 HZ, JH ꢁH = 2.80 HZ, EHC=CH), 2.54
E
E
Z
(unresolved dd centered at, 2H, COCH2S), 2.26 (t, 2H,
J = 7.62 HZ, CH2CO), 2.04 (m, 2H, CH2CH2CO), 1.66
(m, 2H,=CHCH2), 1.65 (m, 2H,=CHCH2CH2), 1.28 (br.s,
8H, CH2(CH2)4); 13C NMR (100 MHZ, CDCl3, d ppm):
169.8, 139.8, 123.3, 114.1, 77.3, 77.2, 77.0, 76.7, 34.1,
33.8, 30.9, 29.3, 25.3, 22.8, 14.2; MS (ESI): m/z =
417.021 [M ? Na]?, Calculated = 417.926.
The fatty acid hydrazides (1a–e) were prepared from the
literature reported method (Rauf et al., 2007).
Synthesis of 3-substituted-4-thiazolidinone (2a–e)
The fatty acid hydrazide (1a–e) (0.1 mmol) and 4-chlo-
robenzaldehyde (0.1 mmol) were refluxed in dry benzene
for 10 h and collecting the generated water in an azeo-
tropic collector, progress of the reaction was monitored
by TLC. The solvent was removed under reduced pres-
sure and solid left was dissolved in N,N-dimethyl
formamide (15–20 ml) then added 0.1 mmol thioglycolic
acid (mercapto acetic acid) and refluxed for 15 h in the
3-(Amido octadec-9-enyl)-2-[40-chloro phenyl]-4-
thiazolidinone (2c)
Light yellow powder; Yield: 76 %, m.p. 124–125 °C;
IR (KBr): 3223 (N–H amide), 2922 (C–H asymm.), 2851
(C–H symm.), 1663 (C=O amide), 1599 (N–H bending),
123

Med Chem Res
Synthesis of 3-substituted-5-methyl-4-
1548 (C=C aromatic), 1116 (C–C aliphatic), 605 (C–Cl
1
aromatic ring substition); H NMR (400 MHZ, CDCl3, d,
thiazolidinone (3b)
ppm): 8.80 (s, 1H, NH), 7.83 (d, 2H, J = 8.56 HZ, Ar 2H),
7.52 (d, 2H, J = 8.40 HZ, Ar 2H), 5.37 (s, 1H, NCHS),
2.74 (unresolved dd centered at, 2H, COCH2S), 2.54 (t, 2H,
J = 7.54 HZ, CH2CO), 1.70 (m, 2H, CH2CH2CO), 1.32
(br.s, 10H, (CH2)5), 1.30 (m, 2H, CH3CH2), 0.87 (dt, 3H,
CH2CH3); 13C NMR (100 MHZ, CDCl3, d, ppm): 168.7,
138.9, 123.1, 115.0, 77.3, 77.2, 77.1, 76.5, 34.0, 30.3,
25.2, 23.1, 14.1; MS (ESI): m/z = 515.011 [M ? Na]?,
Calculated = 516.101.
The fatty acid hydrazide (0.1 mmol) and 4-chlorobenzal-
dehyde (0.01 mmol) were dissolved in dry benzene and
refluxed for 10 h and collecting the generated water in an
azeotropic collector. After the completion of the reaction,
excess solvent was removed under reduced pressure. Now,
to the reaction mixture added (0.1 mmol) thiolactic acid
(2-mercapto propanoic acid) and 40–50 mL of N,N-dime-
thyl formamide and refluxed in the presence of catalytic
amount of anhydrous zinc chloride for 16 h. The reaction
was monitored with the help of TLC; after the completion
the reaction mixture was cooled to room temperature and
then work up with diethyl ether-water, a brownish oily
liquid was obtained, which was further purified by column
chromatography. The characterization data of compound
3b is given below.
3-[Amido-(9Z, 12R)-12-hydroxyl octadec-9-enyl]-2-[40-
chlorophenyl]-4-thiazolidinone (2d)
Yellow powder; Yield: 65–70 %; m.p. 119–120 °C; IR
(KBr): 3219 (N–H amide), 2922 (C–H asymm.), 2851
(C–H symm.), 1671 (C=O amide), 1599 (N–H bending),
1548 (C=C aromatic), 1188 (C–C aliphatic), 546 (C–Cl
aromatic ring substitution); 1H NMR (400 MHZ, CDCl3, d,
ppm): 8.61 (s, 1H, NH), 7.68 (d, 2H, J = 7.76 HZ, Ar 2H),
7.34 (d, 2H, J = 7.92 HZ, Ar 2H), 5.44 (s, 1H, NCHS),4.16
(m, 2H, CH = CH), 3.41 (m, 1H, CHOH), 2.95 (t, 2H,
J = 7.40 HZ, CH2CO), 2.69 (unresolved dd centered at,
2H, COCH2S), 2.27 (m, 1H, CHOH), 2.12 (m, 4H,
CH2CH=CHCH2), 1.82 (m, 2H, CH2CHOH), 1.46 (br.s,
18H, (CH2)9), 0.87 (dt, 3H, CH2CH3); 13C NMR (100
MHZ, CDCl3, d, ppm) 175.5, 169.7, 145.0, 134.9, 132.4,
129.3, 127.6, 77.7, 77.5, 77.3, 77.0, 40.2, 34.5, 31.3, 28.8,
25.1, 22.0, 13.6; MS (ESI): m/z = 531.091 [M ? Na]?,
Calculated = 532.100.
3-(Amido undec-10-enyl)-2-[40-chlorophenyl]-5-methyl-
4-thiazolidinone (3b)
Oily liquid; Yield: 72 %; IR (KBr): 3222 (N–H amide),
2925 (C–H asymm.), 2854 (C–H, symm.), 1641 (C=O
amide), 1541(N–H bending), 1149 (C–C aliphatic), 665
(C–Cl aromatic ring substitution); 1H NMR (400 MHZ,
CDCl3, d, ppm): 8.79 (s, 1H, NH), 7.81 (d, 2H, J = 8.56
HZ, Ar 2H), 7.38 (d, 2H, J = 8.52 HZ, Ar 2H), 5.27 (s, 1H,
NCHS), 5.17 (tdd, 1H, JHꢁ CH
9
=5.80 HZ, J
= 9.10
H–Hz
2
HZ, JHꢁH =15.40 HZ, H2C = CH), 5.01 (dd, 1H,
E
J
H ꢁH=9.10 HZ, JH ꢁH =1.20 HZ, ZHC = CH), 4.15 (dd,
Z
Z
E
1H, JH ꢁH=15.40 HZ, JH ꢁH = 1.20 HZ, HC = CH), 3.08
E
E
E
Z
(q, 1H, J = 6.42 HZ, COCHS), 2.23 (t, 2H, J = 5.06 HZ,
CH2CONH), 1.93 (d, 3H, J = 5.44 HZ, CHCH3), 1.77
(m, 2H, CH2CH2CO), 1.59 (m, 2H, = CH2CH2), 1.33
(m, 2H, = CHCH2CH2), 1.25 (br.s, 8H, CH2(CH2)4); 13C
NMR (100 MHZ, CDCl3, d, ppm): 170.5, 138.1, 130.9,
129.4, 128.8, 77.3, 77.0, 76.7, 68.1, 38.7, 31.8, 30.3, 29.7,
22.6, 14.1, 10.9; MS (ESI): m/z = 431.024 [M ? Na]?,
Calculated = 431.925.
3-[Amido-(9R, 12Z)-9-hydroxyl octadec-12-enyl]-2-[40-
chlorophenyl]-4-thiazolidinone (2e)
Yellow powder; Yield: 72 %; m.p. 121–122 °C; IR
(KBr): 3224 (N–H amide), 2923 (C–H asymm.), 2852
(C–H symm.), 1673 (C=O amide), 1600 (N–H bending),
1544 (C=C aromatic), 1186 (C–C aliphatic), 557 (C–Cl
1
aromatic ring substitution); H NMR (400 MHZ, CDCl3,
d, ppm): 8.63 (s, 1H, NH), 7.79 (d, 2H, J = 8.52 HZ, Ar
2H), 7.43 (d, 2H, J = 8.52 HZ, Ar 2H), 5.43 (s, 1H,
NCHS), 4.11 (m, 2H, CH = CH), 3.71 (m, 1H, CHOH),
2.82 (t, 2H, J = 7.40 HZ, CH2CO), 2.63 (unresolved dd
centered at, 2H, COCH2S), 2.28 (m, 1H, CHOH), 2.17
(m, 4H, CH2CH = CHCH2), 1.70 (m, 2H, CH2CHOH),
1.25 (br.s, 18H, (CH2)9), 0.87 (dt, 3H, CH2CH3); 13C
NMR (100 MHZ, CDCl3, d, ppm): 175.4, 169.7, 145.0,
135.0, 132.3, 129.3, 127.6, 77.6, 77.5, 77.3, 77.0,
40.1, 34.4, 28.8, 22.0, 13.9; MS (ESI): m/z = 532.002
[M ? Na]?, Calculated = 532.100.
Pharmacology
Antibacterial studies
The newly synthesized compounds were screened for their
antibacterial activity against Pseudomonas aeruginosa
(ATCC-27853), Escherichia coli (ATCC-25922), Strepto-
coccus pyogenes and Klebsiella pneumonia (Clinical iso-
lates), Methicillin resistant Staphylococcus aureus (MRSA
?ve), bacterial strains by disk diffusion method (Collins,
1976; Khan, 1997). A sterile glass spreader was used for
123

Med Chem Res
even distribution of the inoculums, a standard inoculums
(1–2 9 107 c.f.u./mL 0.5 McFarland standards) was
introduced onto the surface of sterile agar plates. 6 mm
diameter disks were prepared from Whatman no. 1 filter
paper and that was sterilized by dry heat at 140 °C for 1 h.
The sterile disks were placed in a nutrient agar medium
that were previously soaked in a known concentration of
the test compounds. Positive control used was chloram-
phenicol (30 lg); the structure of the chloramphenicol is
given below. While the disk poured in DMSO was used as
negative control. The plates were inverted and incubated
for 24 h at 37 °C. The susceptibility was estimated on the
basis of the diameter of zone of inhibition against Gram-
positive and Gram-negative strains of bacteria; inhibition
zones were measured and compared with the controls. The
bacterial zones of inhibition values are given in Table 1.
The broth dilution technique was used to determine
minimum inhibitory concentrations (MICs). The nutrient
broth, which contained logarithmic serially twofold diluted
amount of test compound and controls were inoculated
with approximately 5 9 105 c.f.u./mL of actively dividing
bacteria cells. The cultures were incubated for 24 h at
37 °C and the growth was monitored visually and spec-
trophotometrically. The MIC was the lowest concentration
(highest dilution) required to arrest the growth of bacteria.
To obtain the minimum bacterial concentration (MBC),
0.1 mL volume was taken from each tube and spread on
agar plates. The number of c.f.u. was counted after 18–24 h
of incubation at 35 °C. MBC was defined as the lowest
drug concentration at which 99.9 % of the inoculums were
killed. The MIC and MBC are given in Table 2.
Antifungal studies
O
OH
N+
The disk diffusion method was also used for screening of
antifungal activity. For assaying the antifungal activity,
Aspergillus fumigates, Trichophyton mentagrophytes,
Penicillium marneffei, Candida albicans were recultured in
DMSO by agar diffusion methods (Varma, 1998; Ahmad
and Khan, 2009). Peptone (1 g), D-glucose (4 g) and agar
(2 g) was dissolved in distilled water (100 mL) to prepare
O
O-
Cl
N
H
Cl
OH
Chloramphenicol
Table 1 Antibacterial activity
of 2,3-disubstituted-
4-thiazolidinones
(2a–e)
Diameter of zone of inhibition (mm).
Gram-positive bacteria Gram-negative bacteria
Compounds
S. Pyogenes
MRSAa
P. aeruginosa
K. pneumoniae
E. coli
2a
20.4 0.3
20.2 0.4
18.1 0.3
17.1 0.2
15.5 0.4
23.0 0.2
–
20.5 0.4
19.7 0.3
17.5 0.2
15.9 0.3
15.4 0.4
22.0 0.2
–
27.2 0.4
22.3 0.2
19.9 0.3
14.1 0.4
13.9 0.6
32.0 0.3
–
21.4 0.4
18.3 0.5
17.1 0.3
14.5 0.4
13.9 0.2
19.0 0.2
–
23.3 0.4
19.8 0.4
17.6 0.2
14.5 0.3
14.3 0.2
27.0 0.2
–
2b
Positive control (standard);
Chloramphenicol and negative
control (DMSO) measured by
the Halo Zone Test (Unit, mm)
2c
2d
2e
a
Methicillin resistant
Staphylococcus aureus
(MRSA ?ve)
Standard
DMSO
Table 2 MIC and MBC results of 2,3-disubstituted-4-thiazolidinones (2a–e) positive control Chloramphenicol
Compounds
Gram-positive bacteria
Gram-negative bacteria
S. Pyogenes
MRSA
P. aeruginosa
K. pneumoniae
E. coli
MIC
MBC
MIC
MBC
MIC
MBC
MIC
MBC
MIC
MBC
2a
12.5
25
25
12.5
25
25
12.5
12.5
50
25
12.5
25
50
12.5
25
25
2b
50
50
25
50
50
2c
25
100
100
100
12.5
50
100
100
100
12.5
100
100
100
25
25
50
25
100
[100
100
12.5
2d
50
50
50
25
100
[100
25
50
2e
25
50
50
50
50
Standard
12.5
6.25
12.5
6.25
6.25
MIC (lg/mL) minimum inhibitory concentration, that is, the lowest concentration of the compound to inhibit the growth of bacteria completely,
MBC (lg/mL) minimum bacterial concentration, that is, the lowest concentration of the compound for killing the bacteria completely
123

Med Chem Res
the Sabourands agar media and adjusting pH to 5.7. Nor-
mal saline was used to make a suspension of spore of
fungal strain for lawning. A loopful of particular fungal
strain was transferred to 3 mL saline to get a suspension of
corresponding species. Twenty milliliters of agar media
was poured into each petri dish. Excess of suspension was
decanted and the plates were dried by placing in an incu-
bator at 37 °C for 1 h using an agar punch, wells were
made and each well was labeled. A control was also pre-
pared in triplicate and maintained at 37 °C for 3–4 days.
The fungal activity of each newly synthesized compound
was compared with greseofulvin as standard drug; structure
of the drug is given below. Inhibition zones were measured
and compared with the controls. The fungal zones of
inhibition values are given in Table 3. The nutrient broth
contained logarithmic serially twofold diluted amount of
test compound and controls was inoculated with approxi-
mately 1.6 9 104–6 9 104 c.f.u./mL. The cultures were
incubated for 48 h at 35 °C and the growth was monitored.
The lowest concentration (highest dilution) required to
arrest the growth of fungus was regarded as MIC. To obtain
the minimum fungicidal concentration (MFC), 0.1 mL
volume, was taken from each tube and spread on agar
plates. The number of c.f.u. was counted after 48 h of
incubation at 35 °C. MFC was defined as the lowest drug
concentration at which 99.9 % of the inoculums were kil-
led. The MIC and MFC are given in Table 4.
Table 4 MIC and MFC of 2,3-disubstituted-4-thiazolidinone (2a–e)
Comp.
CA
AF
TM
PM
MIC MFC MIC MFC MIC MFC MIC MFC
2a
2b
2c
2d
2e
6.25 12.5 6.25
25
25
6.25
12.5
25
25
50
12.5
12.5
25
25
25
12.5 25
12.5 50
12.55
12.5
25
50
50
50
25
25
50
50
50
25
50
25
100
100
25
25
100
25
100
25
25
Standard 6.25 25
12.5
12.5 6.25
12.5
CA Candida albicans, AF Aspergillus fumigatus, TM Trichophyton
mentagrophytes, PM Penicillium marneffei. MIC (lg/mL) minimum
inhibitory concentration, that is, the lowest concentration of the
compound to inhibit the growth of fungus completely; MFC (lg/mL)
minimum fungicidal concentration, that is, the lowest concentration
of the compound for killing the fungus completely
Conclusions
To the best of our knowledge, thiazolidinone derivatives of
fatty acids have been synthesized for the first time. The
thiazolidinone ring is an important pharmacophore in
modern drug discovery. Thus, several substituted 4-thia-
zolidinones have been synthesized from fatty acid hydra-
zides in appreciable yields and these synthesized
compounds were found to be the good antimicrobial
agents. The compound 2a, 2b, and 2c showed good anti-
bacterial activity equivalent to that of standard used
chloramphenicol and compound 2a, 2b, and 2c showed the
excellent antifungal activity against the above-mentioned
fungal strains. The structure activity relationship of the
novel synthetic long chain hydroxyl and non-hydroxyl
fatty acid derivatives of 4-thiazolidinone was studied
by changing the substituents at third position in the
2-(40-chloro phenyl)-4-thiazolidinone of the products. The
antimicrobial results revealed that compounds having
additional functional group and long chain of fatty acid in
the 2-(40-chloro phenyl)-4-thiazolidinone moiety 2 (a–c)
did not show good activity against most of the pathogenic
microbes. Long chain non-hydroxyl fatty acid derivatives
of 4-thiazolidinone 2 (a–c) showed good antimicrobial
Table 3 Antifungal activity of
2,3-disubstituted-
Diameter of zone of inhibition (mm)
4-thiazolidinone (2a–e) Positive
control (greseofulvin) and
negative control (DMSO)
measured by the Halo Zone Test
(Unit, mm)
Compounds
CA
AF
TM
PM
2a
27.3 0.3
27.1 0.4
24.2 0.5
23.7 0.6
21.2 0.4
30.0 0.2
–
22.6 1.2
22.2 0.2
21.4 0.3
20.6 0.2
18.7 0.3
27.0 0.2
–
19.5 0.7
19.2 1.2
17.6 0.5
18.2 0.2
14.8 0.9
24.0 0.3
–
15.2 0.2
14.6 0.3
14.1 0.9
14.4 0.6
13.3 0.2
20.0 0.5
–
2b
2c
2d
CA Candida albicans, AF
Aspergillus fumigatus, TM
Trichophyton mentagrophytes
2e
Standard
DMSO
PM Penicillium marneffei
123

Med Chem Res
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Acknowledgments Two of us, (HV)1 and (AA)1, are thankful to the
DST and CSIR, New Delhi, respectively, for the award of JRF and we
are also thankful to the ICMR for financial support in part by a Grant
from Major Research Project [ICMR-No 80/613/2009-ECD-I (NR)].
Authors are thankful to the Chairman, Department of Chemistry,
Aligarh Muslim University, for providing the necessary research
facilities and also to the SAIF, Punjab University, Chandigarh, for
providing the spectra analyzing techniques.
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