Synthesis and evaluation of in vitro antimicrobial activity of novel 2-[2-(aroyl)aroyloxy]methyl-1,3,4-oxadiazoles

Article abstract of DOI:10.1134/S1068162014030066

Synthetic pathway of the ten novel 2-[2-(aroyl)aroyloxy]methyl-1,3,4- oxadiazoles as new potential antimicrobial agents is illustrated. Intramolecular cyclization of 2-(2-aroylaryloxy) aceto hydrazides to 2-[2-(aroyl)aroyloxy] methyl-1,3,4-oxadiazoles was

Full text of DOI:10.1134/S1068162014030066

ISSN 1068ꢀ1620, Russian Journal of Bioorganic Chemistry, 2014, Vol. 40, No. 3, pp. 330–335. © Pleiades Publishing, Ltd., 2014.
Synthesis and Evaluation of in vitro Antimicrobial Activity
of Novel 2ꢀ[2ꢀ(Aroyl)aroyloxy]methylꢀ1,3,4ꢀOxadiazoles1
V. Girisha, Noor Fatima Khanumb, H. D. Gurupadaswamya, and Shaukath Ara Khanuma,
2
a Department of Chemistry, University of Mysore, Yuvaraja’s College, Mysore, 570005 India
b Department of Food Science and Nutrition, Maharani’s Science College, Mysore, 570006 India
Recevied July 22, 2013; in final form, October 21, 2013
Abstract—Synthetic pathway of the ten novel 2ꢀ[2ꢀ(aroyl)aroyloxy]methylꢀ1,3,4ꢀoxadiazoles as new potenꢀ
tial antimicrobial agents is illustrated. Intramolecular cyclization of 2ꢀ(2ꢀaroylaryloxy) aceto hydrazides to
2ꢀ[2ꢀ(aroyl)aroyloxy]methylꢀ1,3,4ꢀoxadiazoles was achieved with triethyl orthoformate in good yields. The
1
compounds were characterized by IR, H NMR, mass spectra and by means of CHN analysis. The target
compounds were tested for their in vitro antimicrobial activity against representative strains by disc diffusion
method and micro dilution methods. Several compounds showed antimicrobial activity comparable with or
higher than the standard drugs.
Keywords: benzophenones, 1,3,4ꢀoxadiazoles, antimicrobial activity, MIC
DOI: 10.1134/S1068162014030066
2 1
INTRODUCTION
change in the drug molecule, which may not be
detected by chemical methods can be revealed by a
change in the antimicrobial activity and hence microꢀ
biological assays are very useful for resolving doubts
regarding possible changes in the potency of drugs and
their preparation. The microbial assay is based upon
the compulsion of inhibition of growth of microorganꢀ
isms by marked concentration of the synthetic or natꢀ
ural compounds to be examined with that produced by
the known concentration of a standard drug having a
known activity [10].
The problem of increasing antimicrobial resistance
among bacterial and fungal pathogens is of growing
concern to physicians, microbiologists, research sciꢀ
entists and professionals of the pharmaceutical indusꢀ
try [1, 2]. Even though pharmacological industries
have produced a number of new antibiotics in the last
three decades, resistance to these drugs by microorꢀ
ganisms has increased [3]. In general, microorganisms
have the genetic ability to transmit and acquire resisꢀ
tance to drugs, which are utilized as therapeutic agents
[4]. This is a cause for concern, because a number of
patients have suppressed immunity, and due to new
bacterial and fungal strains, which are multiꢀresistant
[5]. Consequently, new infections can occur resulting
in high mortality [6, 7] Therefore, action must be
taken to counter this problem. For example, measures
such as the controlled use of antibiotic, development
of research to a better understanding of the genetic
mechanisms of resistance and to continue studies to
develop new drugs, either synthetic such as sulfonaꢀ
mides nitrofuranes, penicillins, cephalosporins, tetraꢀ
cycline’s macrolides, and oxazolidinones [8, 9], or
natural. The ultimate goal is to offer appropriate and
efficient antimicrobial drugs to the patient.
Compounds containing benzophenone [11, 12]
and 1,3,4ꢀoxadiazole moieties [13, 14] find a unique
place in medicinal chemistry. 1,3,4ꢀOxadiazole is
associated with potent pharmacological activity due to
the presence of toxophoric –N=C–O– linkage. Conꢀ
siderable evidences have been gathered to reveal the
efficiency of 1,3,4–oxadiazole including antimicroꢀ
bial activity [15]. The synthesis of oxadiazoles continꢀ
ues to attract interest, due to its interesting structural
implications of the biological systems. In view of these
observations and our continued interest in the syntheꢀ
sis of biologically active heterocyclic compounds [16],
it was worthwhile to synthesize integrated 1,3,4–oxaꢀ
diazole moiety to a benzophenone framework.
The inhibitors of microorganism growth under
standardized conditions may be utilized in demonꢀ
strating the therapeutic efficacy of drugs. Any subtle
RESULTS AND DISCUSSION
The reaction sequence for the title compounds is
outlined in scheme. Compounds (1a–j) to (3a–j) have
been prepared as previously reported by our group
[16–18]. Compounds (3a–j) with triethyl orthoforꢀ
1
The article is published in the original.
Corresponding author: phone: 9901888755; eꢀmail:
2
shaukathara@yahoo.co.in.
330

SYNTHESIS AND EVALUATION
331
mate underwent intramolecular cyclization, to yield phenyl ring and the meta position in benzoyl ring,
substituted 2ꢀ[2ꢀaroylaroyloxymethyl]ꢀ1,3,4ꢀoxadiaꢀ respectively showed good activity against both Gramꢀ
zoles (4a–j). The antimicrobial activities of syntheꢀ positive and Gramꢀnegative bacteria among all synꢀ
sized compounds were screened against eight bacteria thesized compounds compared with the standard.
and four fungi using in vitro disc diffusion method. Among the compounds (4g–j) in which chloro group
The results revealed that most of the synthesized comꢀ is substituted in the phenyl ring compounds (4g), (4h
pounds exhibited antimicrobial activities against Staꢀ and (4j) show good activity against S. aureus. Comꢀ
phylococcus aureus St. aureus (MRSA), Enterobacter pound (4a) showed significant antifungal activity
aerogenes Micrococcus luteus Klebsiella pneumonia against B. cinerea and C. krusei. In contrast, comꢀ
Salmonella typhimurium S. paratyphiꢀB Proteus vulꢀ pounds (4b) and (4i) with bromo and (4c) with methꢀ
garis Candida albicans Botyritis cinerea Malassesia oxy groups exhibited lowest activity and this can be
)
,
,
,
,
,
,
,
,
,
pachydermatis, and C. krusei organisms. The results attributed to the bulkiness of bromo and methoxy
are summarized in Table 1 and 2. Compounds (4a), groups which might render the molecule to penetrate
(4d), (4g), (4h), and (4j) showed good activity more through the cell wall. The MIC values of active comꢀ
than standard drug against S. aureus. Compound 4a pounds (4a), (4d–h) and (4j) against bacteria and
with methyl and chloro groups at the para position in fungi are given in Table 3 and 4 respectively.
R2
R2
R3
R1
R3
R1
ClCH COOC H
2 5
2
K CO /Acetone
2
3
O
O
O
OH
O
O
R
R
1a–j
2a–j
R2
R2
R3
R1
R1
R3
N
H NꢀNH H O
2
HC(OEt)
N
2 2
3
C H OH
5
2
H N
O
O
2
O
NH
O
O
O
R
R
3a–j
4a–j
a
b
: R = CH3, R1 = R3 = H, R2 = Cl
: R = CH3, R1 = Br, R2 = R3 = H
c
: R = CH3, R1 = R2 = H, R3 = OCH3
: R = CH3, R1 = R2 = R3 = H
: R = CH3, R1 = R2 = H, R3 = Cl
: R = R3 = CH3, R1 = R2 = H
d
e
f
g
h
i
j
: R = R2 = Cl, R1 = R3 = H
: R = Cl, R1 = R2 = R3 = H
: R = Cl, R1 = Br, R2 = R3 = H
: R = Cl, R1 = R2 = H, R3 = CH3
Scheme 1. Synthesis of 2[2ꢀ(aroyl)aroyloxy]methylꢀ1,3,4ꢀoxadiazoles.
Significant MIC values were observed against
(4d), the presence of chloro group in the benzoyl ring
Gramꢀpositive and Gramꢀnegative bacteria. Comꢀ in compounds (4a) and (4e) increased the potency
pounds (4a), (4e), (4g), (4h) and (4j) showed good against S. aureus by one fold. Interestingly the presꢀ
activity against S. aureus. In comparison to compound ence of chloro group in the phenyl ring in compounds
RUSSIAN JOURNAL OF BIOORGANIC CHEMISTRY Vol. 40
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2014

332
GIRISH et al.
Table 1. In vitro antibacterial activity of compounds (4a–j
)
Zone of inhibition in mm
gramꢀpositive bacteria
gramꢀnegative bacteria
Compounds
S. aureus
(MRSA)
S. aureus
E. aerogenes M. luteus K. pneumonia S. typhimurium S. paratyphiꢀB P. vulgaris
(4a)
19
10
11
18
16
15
23
21
12
23
17
17
9
22
9
23
10
10
14
14
13
19
18
11
16
26
25
11
9
26
10
10
16
13
17
20
20
12
23
25
29
9
27
9
(4b)
(4c)
9
9
7
11
11
10
12
16
13
9
(4d)
9
11
14
15
13
17
10
18
24
14
14
11
13
11
10
14
22
10
12
9
(4e)
13
11
14
13
10
15
22
(4f)
(4g)
11
9
(4h)
(4i)
7
(4j)
9
17
24
Streptomycin
19
(
4g), (4h) and (4j) increased the potency against plated with visualization by UVꢀlight. Melting points
S. aureus by two fold. In comparison to compound 4d were determined on a Thomas Hoover capillary meltꢀ
in 4a the potency is increased by two fold against bacꢀ ing point apparatus with a digital thermometer. IR
teria S. paratyphiꢀB, three fold by S. aureus (MRSA) spectra were recorded in Nujol on FTꢀIR Shimadzu
and S. typhimurium, four fold by M. luteus and K pneuꢀ 8300 spectrometer and 1H NMR spectra were
monia and fivefold by E. aerogenes. Besides, the recorded on a Bruker 400 MHz spectrometer in
potency of compound (4a) is increased by two fold CDCl3. Chemical shifts were recorded in parts per
against fungi B. cinerea, three fold by C. krusei and four million downfield from tetramethylsilane. Mass specꢀ
fold by C. albicans compared to compound (4d). In tra were obtained with a VG70ꢀ70H mass spectromeꢀ
general, compound (4a) showed better activity than ter and the elemental analysis (C, H, and N) was perꢀ
standard drugs for most of the tested bacteria and formed on Elementar Vario EL III elemental analyzer.
fungi.
The synthesis of the hitherto unreported title comꢀ
pounds is as outlined in scheme in 65–73.5% yield.
Hydroxy benzophenones (1a–j) on reaction with
ethyl chloroacetate affords ethyl (2ꢀaroylaryloxy)aceꢀ
EXPERIMENTAL
tates (2a–j) in excellent yield, which on treatment
with hydrazine hydrate yields corresponding 2ꢀ(2ꢀ
Chemicals were purchased from Aldrich Chemical
Co. TLC was performed on aluminumꢀbacked silica
aroylaryloxy)acetohydrazides
(3a–j
)
[16–18].
Intramolecular cyclization of (3a–j) with triethyl
orthoformate resulted substituted 2ꢀ[2ꢀaroylaroyꢀ
loxymethyl]ꢀ1,3,4ꢀoxadiazoles (4a–j).
Table 2. In vitro antifungal activity of compounds 4a–j
Zone of inhibition in mm
Comꢀ
Synthesis of 2ꢀ[2ꢀ(3ꢀchlorobenzoyl)ꢀ4ꢀmethylpheꢀ
noxy]methylꢀ1,3,4ꢀoxadiazole (4a): A suspension of
compound (3a) (0.52 g, 1.6 mmol) in triethyl orthoꢀ
formate (10 mL) was refluxed until (3a) disappeared.
A solid was separated on cooling, which was filtered
off, dried and recrystallized from ethanol to afford
compound (4a). Compounds (4b–h) were synthesized
analogously starting with derivatives (3b–h), respecꢀ
tively.
pounds
C. albicans B. cinerea M. pachydermatis C. krusei
(4a)
(4b)
(4c)
(4d)
(4e)
(4f)
(4g)
(4h)
(4i)
19
9
9
11
9
15
11
11
10
13
22
16
11
7
10
10
11
9
12
9
12
10
20
10
10
12
13
13
12
11
10
11
26
22
12
8
14
14
12
10
13
7
(4
a): Yield 72%. Mp 116–118
°
C; IR (Nujol): 1648
2.3
1
(C=N), 1665 cm–1 (C=O); H NMR (CDCl3):
δ
(3 H, s, CH3), 4.55 (2 H, s, CH2), 6.85–7.7 (7 H, m,
ArꢀH), 9.4 (1 H, s, oxadiazoleꢀH). MS: 328.5
+, 78), 301.5 (49.5), 299.5 (52.5), 285.5 (36.5), 139.5
(4j)
Ketoꢀ
15
16
m/z
conazole
(M
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SYNTHESIS AND EVALUATION
333
Table 3. MIC (
µ
g/mL) of compounds 4a–j against tested bacteria
Minimum inhibitory concentration (
gramꢀpositive bacteria
µ
g/mL)
Comꢀ
gramꢀnegative bacteria
pounds
S. aureus
(MRSA)
S. aureus
E. aerogenes M. luteus K. pneumonia S. typhimurium S. paratyphiꢀB P. vulgaris
(4a)
(4d)
(4e)
(4f)
(4g)
(4h)
(4j)
31.25
62.5
62.5
500
15.62
500
15.62
250
63.5
250
15.62
250
63.5
500
15.62
125
250
62.5
125
500
125
500
500
500
500
<15.62
500
31.25
250
125
125
250
250
250
500
500
15.62
15.62
15.62
6.25
250
32.5
62.5
250
550
250
31.25
31.25
15.62
30
62.5
125
250
62.5
125
62.5
25
125
6.25
125
Streptoꢀ
mycin
>100
6.25
ni
6.25
Ciproꢀ
<0.78
>100
<0.78
>100
<0.78
>100
6.25
<0.78
floxacin
ni = no inhibition.
(100), 111.5 (13.5). Anal. calcd. for C17H13ClN2O3 (CDCl3): δ 2.25 (3 H, s, CH3), 4.53 (2 H, s, CH2),
(328.5): C, 62.11; H, 3.99; Cl, 10.78; N, 8.52. Found: 6.8–7.8 (7 H, m, ArꢀH), 9.36 (1 H, s, oxadiazoleꢀH);
MS: m/z 328.5 (M
+, 76.5), 301.5 (46.5), 299.5 (50.5),
C, 62.09; H, 3.95; Cl, 10.75; N, 8.50%.
285.5 (35.5), 139.5 (100), 111.5 (12.5). Anal. calcd. for
C17H13ClN2O3(328.5): C, 62.11; H, 3.99; Cl, 10.78;
N, 8.52. Found: C, 62.06; H, 3.91; Cl, 10.72; N,
8.51%.
2ꢀ[2ꢀ(2ꢀBromobenzoyl)ꢀ4ꢀmethylphenoxy]methylꢀ
1,3,4ꢀoxadiazole (4b). Yield 73%. Mp 131–133
°
C; IR
(Nujol): 1655 (C=N), 1674 cm–1 (C=O); H NMR
(CDCl3): 2.3 (3 H, s, CH3), 4.51 (2 H s, CH2),
6.8–7.75 (7 H, m, ArꢀH), 9.3 (1 H, s, oxadiazoleꢀH);
MS: 373 (
+, 77), 345 (49), 346 (53), 330 (37),
1
δ
2ꢀ[2(4ꢀMethylbenzoyl)ꢀ4ꢀmethylphenoxy]methylꢀ
m/
z
M
1,3,4ꢀoxadiazole (4f). Yield 72%. Mp 120–122
°
C; IR
(Nujol): 1650 (C=N), 1670 cm–1 (C=O); H NMR
(CDCl3): 2.32 (6 H, s, 2CH3), 4.55 (2 H, s, CH2),
6.85–7.7 (7 H, m, ArꢀH), 9.4 (1 H, s, oxadiazoleꢀH);
1
184 (100), 157 (13). Anal. calcd. for C17H13BrN2O3
(373): C, 54.69; H, 3.48; Br, 21.44, N, 7.50. Found:
C, 54.67; H, 3.45; Br, 21.84, N, 7.30%.
δ
2ꢀ[2ꢀ(4ꢀMethoxybenzoyl)ꢀ4ꢀmethylphenoxy]meꢀ
thylꢀ1,3,4ꢀoxadiazole (4c). Yield 73%. Mp 135–138
IR(Nujol): 1650 (C=N), 1670 cm–1 (C=O); 1H NMR
(CDCl3): 2.25 (3 H s, CH3), 3.8 (3 H, s, OCH3), 4.5
(2 H, s, CH2), 6.9–7.8 (7 H, m, ArꢀH), 9.35 (1 H, s,
oxadiazoleꢀH); MS: 324 (
+, 77.5), 297 (49.5),
295 (54), 281 (38), 135 (100), 107 (14). Anal. calcd. for
C18H16N2O4 (324): C, 66.66; H, 4.97; N, 8.64. Found:
C, 66.63; H, 4.92; N, 8.61%.
°C;
Table 4. MIC (
fungi
µ
g/mL) of compounds 4a–j against tested
δ
Minimum inhibitory concentration (
µ
g/mL)
krusei
31.5
Comꢀ
pounds
m/z
M
M. pachyꢀ
dermatis
C. albicans B. cinerea
C
.
(4a)
31.5
500
125
500
250
500
500
250
250
ni
250
250
125
250
250
500
500
12.5
(4d)
(4e)
(4f)
(4g)
(4h)
(4j)
250
2ꢀ(2ꢀBenzoylꢀ4ꢀmethylphenoxy)methylꢀ1,3,4ꢀoxꢀ
adiazole (4d). Yield 73.5%. Mp 112–114
°
C; IR (Nujol):
1653 (C=N), 1675 cm–1 (C=O); H NMR (CDCl3):
2.28 (3 H, s, CH3), 4.52 (2 H, s, CH2), 6.85–7.8
(8 H, m, ArꢀH), 9.3 (1 H, s, oxadiazoleꢀH); MS:
294 (
+, 76), 267 (47), 265 (52), 251 (35), 105
250
125
125
125
250
250
250
1
δ
500
500
m/
z
M
250
(100), 77 (12). Anal. calcd. for C17H14N2O3 (294): C,
69.38; H, 4.79; N, 9.52. Found: C, 69.31; H, 4.72; N,
9.48%.
Flucoꢀ
nazole
>100
12.5
Ketocoꢀ
nazole
25
25
15
15
2ꢀ[2(4ꢀChlorobenzoyl)ꢀ4ꢀmethylphenoxy]methylꢀ
1,3,4ꢀoxadiazole (4e). Yield 71%. Mp 118–120
°C; IR
1
(Nujol): 1655 (C=N), 1673 cm–1 (C=O); H NMR
ni = no inhibition.
No. 3
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334
GIRISH et al.
MS: m/z 308 (M
+, 78), 280 (50), 281 (53), 265 (38),
119 (100), 92 (14). Anal. calcd. for C18H16N2O3 (308):
C, 70.12; H, 5.19; N, 9.09. Found: C, 70.15; H, 5.15;
N, 9.07%.
Bacterial inoculums were prepared by growing cells
in Mueller Hinton Broth (MHA) (Himedia) for 24 h
at 37°C. These cell suspensions were diluted with sterile
MHB to provide initial cell counts of about 104 CFU/ml.
The filamentous fungi were grown on sabouraud dexꢀ
2ꢀ[2(3ꢀChlorobenzoyl)ꢀ4ꢀchlorophenoxy]methylꢀ
trose agar (SDA) slants at 28°C for 10 days and the
spores were collected using sterile doubled distilled
water and homogenized.
Antibacterial activity was carried out using a disc
diffusion method [20]. Petri plates were prepared with
20 mL of sterile Mueller Hinton Agar (MHA) (Himeꢀ
dia, Mumbai). The test cultures were swabbed on the top
of the solidified media and allowed to dry for 10 min. The
1,3,4ꢀoxadiazole (4g). Yield 68%. Mp 124–126
°
C; IR
(Nujol): 1635 (C=N), 1655 cm–1 (C=O); H NMR
(CDCl3): 4.55 (2 H, s, CH2), 6.85–7.7 (7 H, m, ArꢀH),
9.4 (1 H, s, oxadiazoleꢀH); MS: 349 (
+, 78.5),
1
δ
m/
z
M
322 (50), 320 (53), 306 (38), 139.5 (100), 111.5 (14).
Anal. calcd. for C16H10Cl2N2O3 (349): C, 55.04;
H, 2.89; Cl, 20.31; N, 8.02. Found: C, 55.01; H, 2.85;
Cl, 20.28; N, 8.05%.
tests were conducted at 1000
µ
g/disc. The loaded
2ꢀ(2ꢀBenzoylꢀ4ꢀchlorophenoxy)methylꢀ1,3,4ꢀoxaꢀ
discs were placed on the surface of the medium and
left for 30 min at room temperature for compound difꢀ
fusion. Negative control was prepared using respective
diazole (4h). Yield 70%. Mp 126–128
°
C; IR (Nujol):
1640 (C=N), 1660 cm–1 (C=O); H NMR (CDCl3):
4.55 (2 H, s, CH2), 6.8–7.7 (7 H, m, ArꢀH), 9.2
(1 H, s, oxadiazoleꢀH); MS: 314.5 (
+, 79),
1
solvent. Streptomycin (10
control. The plates were incubated for 24 h at 37
bacteria and 48 h at 27 for fungi. Zone of inhibition
µg/disc) was used as positive
δ
°C
for
m/z
M
°
C
287.5 (51), 285.5 (54), 271.5 (39), 105 (100), 77 (15).
Anal. calcd. for C16H11ClN2O3 (314.5): C, 61.06;
H, 3.52; Cl, 11.26; N, 8.90. Found: C, 61.02; H, 3.56;
Cl, 11.21; N, 8.96%.
was recorded in millimeters and the experiment was
repeated twice.
Minimum inhibitory concentration (MIC) studies
of synthesized compounds were performed according to
the standard reference method for bacteria [21] and filaꢀ
2ꢀ[2(2ꢀBromobenzoyl)ꢀ4ꢀchlorophenoxy]methylꢀ
1,3,4ꢀoxadiazole (4i). Yield 65%. Mp 100–102
IR (Nujol): 1655 (C=N), 1675 cm–1 (C=O); 1H NMR
(CDCl3): 4.5 (2 H, s, CH2), 6.9ꢀ7.65 (7 H, m, ArꢀH),
9.1 (1 H, s, oxadiazoleꢀH); MS: 393.5 (
+, 77),
°C;
mentous fungi. Required concentrations (1000
500 g/mL, 250 g/mL, 125 g/mL, 62.5
31.25 g/mL and 15.62 g/mL) of the compound was
µ
g/mL,
µ
µ
µ
µ
g/mL,
δ
µ
µ
m/z
M
dissolved in DMSO (2%), and diluted to give serial
twoꢀfold dilutions that were added to each medium in
96 well plates. An inoculum of 100 mL from each well
was inoculated. The antiꢀfungal agents ketoconazole,
fluconazole for fungi and streptomycin, ciprofloxacin
for bacteria were included in the assays as positive conꢀ
trols. For fungi, the plates were incubated for 48–72 h
366.5 (50), 364.5 (53), 350.5 (38), 184 (100), 157 (16).
Anal. calcd. for C16H10BrClN2O3 (393.5): C, 48.82; H,
2.56; Br, 20.30; Cl, 9.01; N, 7.12. Found: C, 48.72; H,
2.48; Br, 20.39; Cl, 9.11; N, 7.06%.
2ꢀ[2(4ꢀMethylbenzoyl)ꢀ4ꢀchlorophenoxy]methylꢀ
1,3,4ꢀoxadiazole (4j). Yield 65%. Mp 138–140
°
C; IR
(Nujol): 1630 (C=N), 1660 cm–1 (C=O); H NMR
(CDCl3): 2.2 (3 H, s, CH3), 4.6 (2 H, s, CH2), 6.9–7.6
(7 H, m, ArꢀH), 9.2 (1 H, s, oxadiazoleꢀH); MS: 328.5
+, 75), 301.5 (47), 299.5 (51), 285.5 (35), 119 (100),
1
at 28
24 h at 37
°
C
and for bacteria the plates were incubated for
. The MIC for fungi was defined as the
°C
δ
lowest extract concentration, showing no visible funꢀ
gal growth after incubation time. 5 mL of tested broth
was placed on the sterile MHA plates for bacteria and
incubated at respective temperatures. The MIC for
bacteria was determined as the lowest concentration of
the compound inhibiting the visual growth of the test
cultures on the agar plate.
m/z
(M
92 (12). Anal. calcd. for C17H13ClN2O3 (328.5): C, 62.11;
H, 3.99; Cl, 10.78; N, 8.52. Found: C, 62.05; H, 3.88; Cl,
10.68; N, 8.47%.
Materials and methods for the antimicrobial activꢀ
ity: Streptomycin and ciprofloxacin (Sigma) were used
as positive controls against bacteria. Fluconazole and
ketoconazole (Himedia, Mumbai) were used as posiꢀ
tive controls against fungi.
The following Gramꢀpositive bacteria were used
for the experiments; S. aureus (MTCC 7443), S. aureus
(MRSA) (MTCC 84), E. aerogenes (MTCC 111), M. luꢀ
teus (MTCC 1538). The Gramꢀnegative bacteria inꢀ
cluded, K. pneumonia (MTCC 109), S. typhimurium
(MTCC 2488), S. paratyphiꢀB (MTCC 733), P. vulgaris
(MTCC 321). In addition, fungi C. albicans (MTCC
227), B. cinerea (MTCC 2880), M. pachydermatis,
C. krusei (MTCC 231) were also used for the experiꢀ
ments. All cultures were obtained from the Departꢀ
ment of Microbiology, Manasagangotri, Mysore.
ACKNOWLEDGMENTS
One of the authors Dr. Shaukath Ara Khanum
thanks the University Grants Commission, New
Delhi, India for the grant of Major research Project
[UGC No. F.39ꢀ737/2010 (SR) dated 06.01.2011].
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