Synthesis and biological evaluation of some pyrazolylpyrazolines as anti-inflammatory-antimicrobial agents

Article abstract of DOI:10.1016/j.ejmech.2010.01.059

A new series of pyrazolylpyrazolines (5a-k) was synthesized by the reaction of appropriate chalcones (3a-k) with 4-hydrazinobenzenesulfonamide hydrochloride (4) in ethanol. All the newly synthesized target compounds (5a-k) were screened for their anti-inflammatory activity using carrageenan-induced rat paw edema assay. Compounds 5g and 5j showed pronounced anti-inflammatory activity comparable to the reference standard nimesulide, whereas, compounds 5b, 5d and 5h displayed good anti-inflammatory activity. Additionally, the synthesized compounds were evaluated for their in vitro antimicrobial activity against two Gram-positive bacteria and two Gram-negative bacteria. Four compounds 5c, 5h-5j showed good broad spectrum activity against all the tested Gram-positive and Gram-negative bacterial strains. Compound 5j could be identified as the most biologically active member within this study with an interesting dual anti-inflammatory and antibacterial profile.

Full text of DOI:10.1016/j.ejmech.2010.01.059

European Journal of Medicinal Chemistry 45 (2010) 2650–2655
Contents lists available at ScienceDirect
European Journal of Medicinal Chemistry
journal homepage: http://www.elsevier.com/locate/ejmech
Short communication
Synthesis and biological evaluation of some pyrazolylpyrazolines as anti-
inflammatory–antimicrobial agents
,
a
a
b
b
Pawan K. Sharma a , Satish Kumar , Pawan Kumar , Pawan Kaushik , Dhirender Kaushik ,
*
Yogita Dhingra c, Kamal R. Aneja c
a Department of Chemistry, Kurukshetra University, Kurukshetra-136 119, Haryana, India
b Institute of Pharmaceutical Sciences, Kurukshetra University, Kurukshetra-136 119, India
c Department of Microbiology, Kurukshetra University, Kurukshetra-136 119, India
a r t i c l e i n f o
a b s t r a c t
Article history:
A new series of pyrazolylpyrazolines (5a–k) was synthesized by the reaction of appropriate chalcones
(3a–k) with 4-hydrazinobenzenesulfonamide hydrochloride (4) in ethanol. All the newly synthesized
target compounds (5a–k) were screened for their anti-inflammatory activity using carrageenan-induced
rat paw edema assay. Compounds 5g and 5j showed pronounced anti-inflammatory activity comparable
to the reference standard nimesulide, whereas, compounds 5b, 5d and 5h displayed good anti-
inflammatory activity. Additionally, the synthesized compounds were evaluated for their in vitro anti-
microbial activity against two Gram-positive bacteria and two Gram-negative bacteria. Four compounds
5c, 5h–5j showed good broad spectrum activity against all the tested Gram-positive and Gram-negative
bacterial strains. Compound 5j could be identified as the most biologically active member within this
study with an interesting dual anti-inflammatory and antibacterial profile.
Received 21 October 2009
Received in revised form
20 January 2010
Accepted 21 January 2010
Available online 29 January 2010
Keywords:
Chalcones
Pyrazolylpyrazolines
Antimicrobial activity
Anti-inflammatory activity
Ó 2010 Elsevier Masson SAS. All rights reserved.
1. Introduction
a potential template for dual antimicrobial–anti-inflammatory
agents. It must be noted that this scaffold provides vicinal diaryl
Five-membered heterocycles with a conserved vicinal 1,2-diaryl
substitution pattern are ideal representatives of a recurring core
structure that is found in numerous biologically active compounds,
including cyclooxygenase inhibitors, kinase inhibitors, GPCR
antagonists and even agonists, phosphatase inhibitors, and dopa-
mine transporter inhibitors [1]. 1,2-Diaryl substituted heterocycle
system occurs in so many diverse classes of biologically interesting
low molecular-weight compounds that it would be an under-
statement to link it to the ease of synthesis of the vicinal diaryl
system [2]. Drugs based on a pyrazole ring bearing two adjacent
aryl groups in a vicinal relation have often been occupying a posi-
tion in the list of best selling pharmaceutical products since the
beginning of this decade [2]. Many pyrazole derivatives have been
reported to possess diverse pharmacological activities such as anti-
inflammatory [3–5], antimicrobial [6–8], antihypertensive [9], etc.
However studies investigating the potential of pyrazole derivatives
as dual antimicrobial–anti-inflammatory agents have only recently
been initiated [10–13]. Appreciation of these findings motivated us
to synthesize a novel series of pyrazolylpyrazolines (5a–5n) as
substitution pattern on both the pyrazole as well as pyrazoline
nucleus.
2. Results and discussion
2.1. Chemistry
The synthetic route used to synthesize the target 4-[5-(1-
phenyl-3-aryl-1H-pyrazol-4-yl)-3-aryl-4,5-dihydro-1H-pyrazol-1-
yl]benzenesulfonamides (5a–k) is outlined in Scheme 1.1-Phenyl-3-
aryl-1H-pyrazole-4-carbaldehydes (1a–d), prepared by the
Vilsemeier–Haack reaction of the corresponding hydrazones [14,15],
were subjected to base catalyzed Claisen–Schmidt condensation
reaction with appropriate acetophenones (2a–2d) generating the
required 1-aryl-3-(1-phenyl-3-aryl-1H-pyrazol-4-yl)prop-2-en-1-
ones (3a–k). Finally pyrazolylpyrazolines (5a–5k) with a conserved
vicinal diaryl substitution pattern bearing benzenesulfonamide
moiety were obtained by the condensation of appropriate chalcones
(3a–3k) and 4-hydrazinobenzenesulfonamide hydrochloride (4) in
ethanol containing catalytic amount of glacial acetic acid. 4-Hydra-
zinobenzenesulfonamide (4) was prepared via diazotization of
sulfanilamide followed by reduction of the corresponding diazo-
nium salt with stannous chloride [16]. The purity of compounds was
* Corresponding author. Tel.: þ91 11 9416457355; fax: þ91 1744 238277.
E-mail address: talk2pawan@gmail.com (P.K. Sharma).
0223-5234/$ – see front matter Ó 2010 Elsevier Masson SAS. All rights reserved.
doi:10.1016/j.ejmech.2010.01.059

P.K. Sharma et al. / European Journal of Medicinal Chemistry 45 (2010) 2650–2655
2651
Ph
N
N
R
R1
SO2NH2
Ph
MeOH/NaOH/THF
N
N
+
+
O
R
COCH3
R1
NHNH2
HCl
CHO
1
2
3
4
R = H, CH3, F, Br
R1 = H, CH3, F, Br
Ph
N
N
H2NO2S
R
C2H5OH/AcOH, reflux, 6-7h
N
N
R1
5
Compound 3,5
a
b
c
d
e
f
g
h
i
j
k
R
H
H
H
H
CH3 CH3 CH3
CH3
F
F
F
Br Br
R1
H
CH3
F
Br
H
F
H
H
F
Scheme 1. Synthesis of 5a–5k from formylpyrazoles (1).
checked by TLC and elemental analysis. Analytical and spectral data
(1H NMR, 13C NMR, IR and mass) of the newly synthesized
compounds were in full agreement with the proposed structures. In
general, the characteristic signals in 1H NMR of target pyr-
azolylpyrazolines are the three pyrazoline protons which displayed
a typical ABX type pattern of doublet of doublet. Methine proton of
doublet at around d 3.30 merging with the water signal from DMSO-
d6 in most of the cases.
2.2. Biological evaluation
2.2.1. In vivo anti-inflammatory activity
pyrazolines resonates at around
coupling constants of nearly 12 Hz and 6 Hz. The two methylene
protons displayed two signals; a doublet of doublet at around 4.00
d 5.65 as a doublet of doublet with
All the newly synthesized pyrazolylpyrazolines (5a–k) were
evaluated for their in vivo anti-inflammatory activity by
carrageenan-induced paw edema method [17]. The protocol of
animal experiments has been approved by the Institutional Animal
d
with coupling constants of nearly 17 Hz and 12 Hz and a doublet of
Table 1
Anti-inflammatory activity (AI) of the compounds 5a–5k in carrageenan-induced rat paw edema assay (acute inflammatory model).
Compounda
Volume of edema (mL)b
0 min
30 min
60 min
90 min
120 min
Control
5a
5b
5c
5d
5e
5f
5g
5h
5i
5j
5k
0.38 ꢀ 0.03
0.32 ꢀ 0.01
0.30 ꢀ 0.01
0.37 ꢀ 0.02
0.34 ꢀ 0.03
0.32 ꢀ 0.02
0.35 ꢀ 0.01
0.36 ꢀ 0.04
0.30 ꢀ 0.01
0.29 ꢀ 0.02
0.34 ꢀ 0.01
0.36 ꢀ 0.04 (5)
0.27 ꢀ 0.10
0.53 ꢀ 0.04
0.67 ꢀ 0.09
0.84 ꢀ 0.04
0.93 ꢀ 0.01
0.51 ꢀ 0.01 (5)c
0.50 ꢀ 0.06 (5)
0.41 ꢀ 0.01* (22)
0.49 ꢀ 0.06 (7)
0.42 ꢀ 0.08 (18)
0.44 ꢀ 0.01 (18)
0.37 ꢀ 0.04** (30)
0.45 ꢀ 0.05 (18)
0.43 ꢀ 0.07 (18)
0.44 ꢀ 0.01 (16)
0.46 ꢀ 0.04 (13)
0.33 ꢀ 0.01** (37)
0.64 ꢀ 0.02 (4)
0.75 ꢀ 0.02 (10)
0.65 ꢀ 0.01**(27)
0.64 ꢀ 0.02** (23)
0.65 ꢀ 0.01** (22)
0.64 ꢀ 0.04** (22)
0.70 ꢀ 0.03 (16)
0.55 ꢀ 0.01** (34)
0.66 ꢀ 0.01** (22)
0.70 ꢀ 0.03 (16)
0.52 ꢀ 0.01** (38)
0.69 ꢀ 0.03* (17)
0.48 ꢀ 0.08** (45)
0.92 ꢀ 0.01 (1)
0.55 ꢀ 0.02* (17)
0.50 ꢀ 0.06** (25)
0.51 ꢀ 0.01** (23)
0.57 ꢀ 0.15 (14)
0.54 ꢀ 0.01** (19)
0.38 ꢀ 0.10** (43)
0.58 ꢀ 0.02 (14)
0.61 ꢀ 0.02 (8)
0.68 ꢀ 0.01**(26)
0.79 ꢀ 0.01* (15)
0.71 ꢀ 0.04** (23)
0.78 ꢀ 0.03* (16)
0.84 ꢀ 0.01 (9)
0.63 ꢀ 0.01** (32)
0.71 ꢀ 0.03** (23)
0.84 ꢀ 0.02 (9)
0.48 ꢀ 0.06** (26)
0.49 ꢀ 0.09** (26)
0.38 ꢀ 0.10** (43)
0.64 ꢀ 0.01** (32)
0.79 ꢀ 0.01* (15)
0.59 ꢀ 0.06** (36)
Nimesulide
*Significantly different compared to respective control values, P < 0.05.
**Significantly different compared to respective control values, P < 0.01.
a
Dose levels: test compounds (50 mg/kg body wt), nimesulide (4 mg/kg body wt).
b
Values are expressed as mean ꢀ SEM and analyzed by ANOVA.
c
Values in parentheses (percentage anti-inflammatory activity, AI%).

2652
P.K. Sharma et al. / European Journal of Medicinal Chemistry 45 (2010) 2650–2655
Table 2
In vitro antibacterial activity of compounds 5a–5k by using agar well diffusion method.
Compound
Diameter of zone of inhibition in mma
S. aureus
B. subtilis
P. aeruginosa
E. coli
5a
5b
5c
5d
5e
5f
5g
5h
17.66 ꢀ 0.57 (32)
16.33 ꢀ 0.57 (32)
23.66 ꢀ 0.57 (8)
17.00 (32)
18.66 ꢀ 0.57 (32)
17.00 (32)
23.33 ꢀ 0.57 (8)
11.33 ꢀ 1.15 (64)
11.00 (64)
10.66 ꢀ 0.57 (64)
16.33 ꢀ 0.57 (32)
11.66 ꢀ 0.57 (16)
10.66 ꢀ 0.57 (16)
23.66 ꢀ 0.57 (8)
11.00 ꢀ 1.00 (64)
29.2 (2)
10.66 ꢀ 0.57 (64)
11.66 ꢀ 0.57 (64)
17.66 ꢀ 0.57 (32)
9.66 ꢀ 0.57 (128)
9.33 ꢀ 0.57 (128)
9.33 ꢀ 0.57 (128)
11.00 (64)
17.33 ꢀ 1.15 (32)
19.33 ꢀ 0.57 (32)
17.33 ꢀ 1.15 (32)
9.33 ꢀ 0.57 (128)
28.0 (4)
11.66 ꢀ 0.57 (64)
11.66 ꢀ 0.57 (64)
21.66 ꢀ 0.57 (16)
11.66 ꢀ 0.57 (64)
11.66 ꢀ 0.57 (64)
10.66 ꢀ 0.57 (64)
11.66 ꢀ 0.57 (64)
17.33 ꢀ 0.57 (32)
10.66 ꢀ 0.57 (32)
17.33 ꢀ 1.15 (32)
9.33 ꢀ 0.57 (128)
28.6 (4)
16.00 (32)
16.66 ꢀ 0.57 (32)
16.66 ꢀ 0.57 (32)
23.33 ꢀ 0.57 (16)
21.00 ꢀ 1.00 (16)
24.00 (8)
5i
5j
5k
10.00 (64)
32.6 (2)
Ciprofloxacin
a
Values in parentheses; MIC (
m
g/mL) ¼ minimum inhibitory concentration.
Ethics Committee (IAEC). Each test compound was dosed orally
(50 mg/kg body weight) 30 min prior to induction of inflammation
by carrageenan injection. Nimesulide was utilized as a reference
anti-inflammatory drug at a dose of 4 mg/kg, i.p. The anti-
inflammatory activity was then calculated 30–120 min after
induction and presented in Table 1 as the mean paw volume (mL) in
addition to the percentage anti-inflammatory activity (AI%).
A comparative study of the anti-inflammatory activity of test
compounds relative to the reference drug at different time intervals
indicated the following: after 1 h, compound 5g was nearly as
effective in inhibiting the paw edema with percentage activity of
43% when compared with that of nimesulide (43%). Five other
compounds 5c, 5d, 5f, 5j and 5k showed distinctive pharmacoki-
netic profiles as revealed from their potent and rapid onset of action
with percentage activity of 19–26%. After 90 min, seven compounds,
5b–5e, 5g–5j showed significant anti-inflammatory activity ranging
from 22 to 38% inhibition as compared to nimesulide (45%). Taking
the anti-inflammatory activity after 2 h time interval as a criterion
for comparison, it can be concluded that compounds 5g and 5j
showed potent anti-inflammatory activity (32%) comparable with
nimesulide (36%), whereas, compounds 5b, 5d and 5h displayed
a good anti-inflammatory activity (23–26%), however, none of them
was found to be superior over the reference drug.
3. Conclusion
The objective of the present study was to synthesize and
investigate the anti-inflammatory and antimicrobial activities of
a new series of pyrazolylpyrazolines with the hope of discovering
new structure leads serving as dual anti-inflammatory–antimi-
crobial agents. The substitution pattern of the pyrazole ring as well
as the pyrazoline ring was rationalized so as to be correlated to the
vicinal diaryl heterocycles template. Amongst the tested
compounds, 5g and 5j showed pronounced anti-inflammatory
activity (32%) that was comparable to nimesulide (36%), whereas,
compounds 5b, 5d and 5h displayed good anti-inflammatory
activity (23–26%). However, none of the newly synthesized
compounds was found to be superior over the reference drug. On
the other hand, some of the newly synthesized pyrazolylpyrazo-
lines containing F or Br as a substituent (5c, 5h–5j) were able to
inhibit the growth of both the Gram-positive as well as Gram-
negative bacteria. Finally compound 5j could be identified as the
most biologically active member within this study with an inter-
esting dual anti-inflammatory and antibacterial profile. Conse-
quently pyrazolylpyrazolines represent a class that needs further
investigation with the hope of finding new dual anti-inflamma-
tory–antimicrobial agents.
2.2.2. In vitro antimicrobial activity
4. Experimental protocols
All the target compounds were evaluated for their in vitro
antimicrobial activity against Staphylococcus aureus (MTCC 3160)
and Bacillus subtilis (MTCC 121) representing Gram-positive
bacteria, and Pseudomonas aeruginosa (MTCC 741) and Escherichia
coli (MTCC 51) representing Gram-negative bacteria (Table 2). The
microdilution susceptibility test in Mu¨ller–Hinton broth was used
for the determination of antibacterial activity. Ciprofloxacin was
used as the reference drug. The results were recorded for each
tested compound as the average diameter of inhibition zones of
bacterial growth around the disks in mm. The minimum inhibitory
concentration (MIC) measurement was determined using a micro-
plate dilution method [18,19] (Table 2).
Results revealed that in general, most of the tested compounds
showed better activity against the Gram-positive rather than the
Gram-negative bacteria. Four compounds 5c, 5h–5j showed good
broad spectrum activity against all the tested Gram-positive and
Gram-negative bacterial strains. Based on these preliminary results,
it can be seen that all the four compounds 5c, 5h–5j showing good
antimicrobial activity have a halogen, F or Br, as one of the
substituents. It can further be noted that the presence of haloge-
nated benzene ring on the pyrazole moiety of the pyrazolylpyr-
azolines is better for activity as compared to the presence of
halogenated benzene ring on the pyrazoline moiety.
Melting points were determined in open glass capillaries in an
electrical melting point apparatus and are uncorrected. The
infrared (IR) spectra were recorded on Shimadzu-21 FT-IR or Per-
kin–Elmer IR Spectrophotometer using the KBr pellet technique. 1H
NMR and 13C NMR spectra were recorded in CDCl3/DMSO-d6 on
a Bruker NMR spectrometer at 300 MHz and 75.5 MHz respectively
using tetramethylsilane (TMS) as internal standard. Chemical shifts
are expressed in d, ppm. Mass spectra were recorded on a Waters
Micromass Q-Tof Micro instrument in ESþ mode. The purity of the
compounds was checked by thin layer chromatography (TLC) on
silica gel plates using a mixture of petroleum ether and ethyl
acetate. Iodine was used as a visualizing agent.
4.1. General procedure for the preparation of chalcones 3a–k
To a cold, stirred mixture of methanol (20 mL) and sodium
hydroxide (12.09 mmol) was added appropriate acetophenone (2,
4.03 mmol). The reaction mixture was stirred for 10 min. To this
was added appropriate formyl pyrazole (1, 4.03 mmol) followed
by tetrahydrofuran (30 mL). The solution was further stirred for
2 h at 0 ꢁC and then at room temperature for 5 h. It was then
poured into ice cold water. The resulting solution was neutralized

P.K. Sharma et al. / European Journal of Medicinal Chemistry 45 (2010) 2650–2655
2653
with dil. HCl. The solid so separated was filtered, washed with
4.1.11. (E)-3-[3-(4-Bromophenyl)-1-phenyl-1H-pyrazol-4-yl]-
water, dried and crystallized from ethanol to afford product
3a–3k (60–80% yield).
1-(4-fluorophenyl)-2-propen-1-one (3k)
m.p. 134–136 ꢁC, yield 65%; IR (KBr) 1661 (s, C]O stretch), 1290
(m, CH]CH) cmꢂ1 1H NMR (CDCl3, 300 MHz):
; d 8.36 (s, 1H,
4.1.1. (E)-3-(1,3-Diphenyl-1H-pyrazol-4-yl)-1-phenyl-2-propen-
1-one (3a)
pyrazole-H), 8.10–7.92 (m, 2H, Ar), 7.87 (d, 1H, J ¼ 15.6 Hz, ]CH),
7.80 (d, 2H, J ¼ 8.7 Hz, Ar), 7.66–7.62 (m, 3H, Ar), 7.57 (d, 2H,
J ¼ 8.7 Hz, Ar), 7.36 (d, 1H, J ¼ 15.6 Hz, ]CH), 7.28–7.16 (m, 4H, Ar).
m.p. 133–134 ꢁC; lit. [20] m.p. 134 ꢁC.
4.1.2. (E)-1-(4-Methylphenyl)-3-(1,3-diphenyl-1H-pyrazol-4-yl)-
2-propen-1-one (3b)
4.2. General procedure for the preparation of
pyrazolylpyrazolines 5a–k
m.p. 158–160 ꢁC, yield 70%; IR (KBr) 1659 (s, C]O stretch), 1292
(m, CH]CH) cmꢂ1 1H NMR (CDCl3, 300 MHz):
; d 8.36 (s, 1H,
Pyrazole-H), 8.03–7.27 (m, 16H, Ar, 2ꢃ ]CH), 2.45 (s, 3H, CH3).
To an acidic solution of appropriate chalcone (3, 2.57 mmol) in
ethanol was added 4-hydrazinobenzenesulfonamide hydrochloride
(4, 2.57 mmol). The reaction mixture was refluxed for 5–6 h, cooled
to room temperature. The solid separated was filtered, dried, and
crystallized from ethanol.
4.1.3. (E)-1-(4-Fluorophenyl)-3-(1,3-diphenyl-1H-pyrazol-4-yl)-
2-propen-1-one (3c)
m.p. 175–176 ꢁC; lit. [18] m.p. 177–178 ꢁC.
4.1.4. (E)-1-(4-Bromophenyl)-3-(1,3-diphenyl-1H-pyrazol-4-yl)-
2-propen-1-one (3d)
4.2.1. 1-(4-Aminosulfonylphenyl)-3-phenyl-5-(1,3-
diphenylpyrazol-4-yl)-2-pyrazoline (5a)
m.p. 157–158 ꢁC; lit. [18] m.p. 158–160 ꢁC.
m.p. 184–188 ꢁC, yield 720 mg (72%); IR (KBr) 3395 and 3267 (m,
N–H stretch), 1591 (s, N–H bend), 1337 and 1153 (s, SO2 stretch)
4.1.5. (E)-3-[3-(4-Methylphenyl)-1-phenyl-1H-pyrazol-4-yl]-
1-phenyl-2-propen-1-one (3e)
cmꢂ1; 1H NMR (DMSO-d6, 300 MHz): 8.34 (s, 1H, C50-H pyrazole),
d
7.83 (d, 2H, J ¼ 8.4 Hz, Ar), 7.77–7.79 (m, 4H, Ar), 7.58 (d, 2H,
J ¼ 8.4 Hz, Ar), 7.54 (d, 2H, J ¼ 7.2 Hz, Ar), 7.52–7.44 (m, 6H, Ar),
7.29–7.24 (m, 1H, Ar), 7.04 (d, 2H, J ¼ 8.4 Hz, Ar), 7.00 (s, 2H, ex,
SO2NH2), 5.68 (dd, 1H, J ¼ 6.6 Hz, 12.3 Hz, C5-H pyrazoline), 4.09
(dd, 1H, J ¼ 12.3 Hz, 17.4 Hz, C4-H pyrazoline), 3.35 (m, 1H, C4-H
pyrazoline merged with peak of HOD); 13C NMR (DMSO-d6,
m.p. 130–134 ꢁC, yield 60%; IR (KBr) 1659 (s, C]O stretch), 1292
(m, CH]CH) cmꢂ1 1H NMR (CDCl3, 300 MHz):
; d 8.35 (s, 1H,
Pyrazole-H), 8.00–7.29 (m, 16H, Ar, 2ꢃ ]CH), 2.42 (s, 3H, CH3).
4.1.6. (E)-1-(4-Methylphenyl)-3-[3-(4-methylphenyl)-1-phenyl-
1H-pyrazol-4-yl]-2-propen-1-one (3f)
75.5 MHz):
d 149.9, 149.8, 146.1, 139.2, 133.3, 132.6, 131.9, 129.5,
m.p. 101–103 ꢁC, yield 62%; IR (KBr) 1659 (s, C]O stretch), 1291
129.3, 128.8, 128.7, 128.1, 127.1, 126.4, 122.1, 118.3, 118.2, 112.2, 55.2,
42.6; Elemental analysis Found: C, 69.04; H, 4.52; N, 13.20%:
C30H25N5O2S requires: C, 69.34; H, 4.85; N, 13.49%; LRMS: m/z, 519
(Mþ); C30H25N5O2S requires: m/z, 519.
(m, CH]CH) cmꢂ1 1H NMR (CDCl3, 300 MHz):
; d 8.33 (s, 1H,
Pyrazole-H), 8.00–7.27 (m, 15H, Ar, 2ꢃ ]CH), 2.43 (s, 3H, CH3), 2.40
(s, 3H, CH3).
4.1.7. (E)-1-(4-Fluorophenyl)-3-[3-(4-methylphenyl)-1-phenyl-
1H-pyrazol-4-yl]-2-propen-1-one (3g)
4.2.2. 1-(4-Aminosulfonylphenyl)-3-(4-methylphenyl)-
5-(1,3-diphenylpyrazol-4-yl)-2-pyrazoline (5b)
m.p. 120–122 ꢁC, yield 70%; IR (KBr) 1659 (s, C]O stretch), 1293
m.p. 180–182 ꢁC, yield 57%; IR (KBr) 3372 and 3271 (m, N–H
(m, CH]CH) cmꢂ1
;
1H NMR (CDCl3, 300 MHz):
d
8.36 (s, 1H,
stretch), 1593 (s, N–H bend), 1338 and 1153 (s, SO2 stretch) cmꢂ1
;
Pyrazole-H), 8.03–7.98 (m, 2H, Ar), 7.91 (d, 1H, J ¼ 15.3 Hz, ]CH),
7.82 (d, 2H, J ¼ 8.1 Hz, Ar), 7.62 (d, 2H, J ¼ 8.1 Hz, Ar), 7.54–7.49 (m,
3H, Ar), 7.39–7.27 (m, 3H, Ar, ]CH), 7.17 (t, 2H, J ¼ 8.4 Hz, Ar), 2.45
(s, 3H, CH3).
1H NMR (DMSO-d6, 300 MHz): 7.88 (s,1H, C50-H pyrazole), 7.75 (d,
d
2H, J ¼ 7.2 Hz, Ar–H), 7.68 (d, 2H, J ¼ 8.1 Hz, Ar), 7.64–7.61 (m, 4H,
Ar), 7.53–7.48 (m, 2H, Ar), 7.45–7.35 (m, 3H, Ar), 7.25–7.18 (m, 3H,
Ar, SO2NH2), 6.82 (d, 2H, J ¼ 8.7 Hz, Ar), 5.58 (dd, 1H, J ¼ 5.7 Hz,
11.7 Hz, C5-H pyrazoline), 3.92 (dd, 1H, J ¼ 12 Hz, 16.8 Hz, C4-H
pyrazoline), 3.37–3.20 (m, 1H, C4-H pyrazoline merged with peak of
HOD), 2.17 (s, 3H, CH3); Elemental analysis Found: C, 69.61; H, 5.04;
N, 13.01%; C31H27N5O2S requires: C, 69.77; H, 5.10; N, 13.12%; LRMS:
m/z, 533 (Mþ); C31H27N5O2S requires: m/z, 533.
4.1.8. (E)-3-[3-(4-Fluorophenyl)-1-phenyl-1H-pyrazol-4-yl]-
1-phenyl-2-propen-1-one (3h)
m.p. 110–112 ꢁC, yield 83%; IR (KBr) 1659 (s, C]O stretch), 1290
(m, CH]CH) cmꢂ1 1H NMR (CDCl3, 300 MHz):.
; d 8.35 (s, 1H,
Pyrazole-H), 8.00–7.30 (m, 14H, Ar, 2ꢃ ]CH), 7.12–7.10 (m, 2H, Ar)
4.2.3. 1-(4-Aminosulfonylphenyl)-3-(4-fluorophenyl)-
5-(1,3-diphenylpyrazol-4-yl)-2-pyrazoline (5c)
4.1.9. (E)-1-(4-Fluorophenyl)-3-[3-(4-fluorophenyl)-1-phenyl-
1H-pyrazol-4-yl]-2-propen-1-one (3i)
m.p. 196–198 ꢁC, yield 81%; IR (KBr) 3393 and 3271 (m, N–H
m.p. 168–170 ꢁC, yield 58%; IR (KBr) 1661 (s, C]O stretch), 1291
stretch), 1593 (s, N–H bend), 1337 and 1153 (s, SO2 stretch) cmꢂ1
;
(m, CH]CH) cmꢂ1
;
1H NMR (CDCl3, 300 MHz):
d
8.35 (s, 1H,
1H NMR (DMSO-d6, 300 MHz): 8.36 (s,1H, C50-H pyrazole), 7.84 (d,
d
Pyrazole-H), 7.99 (dd, 2H, J ¼ 9.0 Hz, J ¼ 5.4 Hz, Ar), 7.86 (d, 1H,
J ¼ 15.6 Hz, ]CH), 7.81–7.78 (m, 2H, Ar), 7.69 (dd, 2H, J ¼ 9.0 Hz,
J ¼ 5.4 Hz, Ar), 7.53–7.48 (m, 3H, Ar), 7.33 (d, 1H, J ¼ 15.6 Hz, ]CH),
7.13–7.19 (m, 4H, Ar).
4H, J ¼ 7.5 Hz, Ar), 7.77 (d, 2H, J ¼ 8.1 Hz, Ar), 7.60 (d, 2H, J ¼ 8.7 Hz,
Ar), 7.55–7.42 (m, 5H, Ar), 7.31–7.28 (m, 3H, Ar), 7.05 (d, 2H,
J ¼ 8.7 Hz, Ar), 7.01 (s, 2H, ex, SO2NH2), 5.70 (dd, 1H, J ¼ 6.3 Hz,
12.0 Hz, C5-H pyrazoline), 4.08 (dd, 1H, J ¼ 12.0 Hz, 17.4 Hz, C4-H
pyrazoline), 3.37–3.24 (m, 1H, C4-H pyrazoline merged with peak of
4.1.10. (E)-3-[3-(4-Bromophenyl)-1-phenyl-1H-pyrazol-4-yl]-
1-phenyl-2-propen-1-one (3j)
HOD); 13C NMR (DMSO-d6, 75.5 MHz):
d
162.6 (d, 1JCF ¼ 246 Hz),
149.8, 148.9, 146.1, 139.1, 133.3, 129.5, 128.7, 128.5, 128.2, 128.1 (d,
2
m.p. 162–164 ꢁC, yield 76%; IR (KBr) 1662 (C]O stretch), 1291
3JCF ¼ 8.5 Hz), 127.1, 126.4, 122.0, 118.2, 115.7 (d, JCF ¼ 21.9 Hz),
(m, CH]CH) cmꢂ1
;
1H NMR (CDCl3, 300 MHz):
d
8.36 (s, 1H,
112.2, 55.2, 42.6. Elemental analysis Found: C, 67.12; H, 4.63; N,
13.22%; C30H24FN5O2S requires: C, 67.02; H, 4.50; N, 13.03%; LRMS:
m/z, 537 (Mþ); C30H24FN5O2S requires: m/z, 537.
pyrazole-H), 7.98–7.78 (m, 5H, Ar, ]CH), 7.64–7.35 (m, 11H,
Ar, ]CH).

2654
P.K. Sharma et al. / European Journal of Medicinal Chemistry 45 (2010) 2650–2655
4.2.4. 1-(4-Aminosulfonylphenyl)-3-(4-bromophenyl)-
5-(1,3-diphenylpyrazol-4-yl)-2-pyrazoline (5d)
4.2.8. 1-(4-Aminosulfonylphenyl)-3-phenyl-5-(3-(4-fluorophenyl)-
1-phenylpyrazol-4-yl)-2-pyrazoline (5h)
m.p. 195–198 ꢁC, yield 62%; IR (KBr) 3368 and 3258 (m, N–H
m.p. 216–218 C, yield 69%; IR (KBr) 3370 and 3269 (m, N–H
stretch), 1595 (s, N–H bend), 1336 and 1152 (s, SO2 stretch) cmꢂ1
;
stretch), 1593 (s, N–H bend), 1337 and 1153 (s, SO2 stretch) cmꢂ1
;
1H NMR (DMSO-d6, 300 MHz):
d
8.36 (s,1H, C50-H pyrazole), 7.83 (d,
1H NMR (DMSO-d6, 300 MHz): 8.36 (s,1H, C50-H pyrazole), 7.82 (d,
d
2H, J ¼ 8.1 Hz, Ar), 7.76–7.70 (m, 4H, Ar), 7.65–7.59 (m, 2H, Ar), 7.56–
7.42 (m, 8H, Ar), 7.04 (d, 2H, J ¼ 9.0 Hz, Ar), 7.00 (s, 2H, ex, SO2NH2),
5.70 (dd, 1H, J ¼ 6.3 Hz, 12.0 Hz, C5-H pyrazoline), 4.07 (dd, 1H,
J ¼ 12.0 Hz, 17.4 Hz, C4-H pyrazoline), 3.42–3.34 (m, 1H, C4-H pyr-
azoline merged with HOD peak); 13C NMR (DMSO-d6, 75.5 MHz):
2H, J ¼ 8.4 Hz, Ar), 7.80–7.75 (m, 4H, Ar), 7.59 (d, 2H, J ¼ 8.7 Hz, Ar),
7.46–7.40 (m, 6H, Ar), 7.35–7.25 (m, 2H, Ar), 7.07–7.04 (d, 2H,
J ¼ 9.0 Hz, Ar), 7.00 (s, 2H, ex, SO2NH2), 5.68 (dd, 1H, J ¼ 6.0 Hz,
17.1 Hz, C5-H pyrazoline), 4.07 (dd, 1H, J ¼ 12.0 Hz, 17.1 Hz, C4-H
pyrazoline), 3.42–3.31 (m, 1H, C4-H pyrazoline merged with HOD
d
149.9, 148.8, 145.9, 139.1, 133.5, 131.6, 131.2, 129.5, 129.4, 128.7,
peak); 13C NMR (DMSO-d6, 75.5 MHz):
d
162.3 (d, 1JCF ¼ 246 Hz),
128.1, 127.2, 127.1, 126.5, 122.4, 121.9, 118.2, 112.3, 55.3, 42.4.
Elemental analysis Found: C, 60.02; H, 4.00; N, 11.54%;
C30H24BrN5O2S requires: C, 60.20; H, 4.04; N, 11.70%; LRMS: m/z,
597, 599 (Mþ); C30H24BrN5O2S requires: m/z, 597, 599
149.7, 148.9, 146.0, 139.0, 133.2, 131.8, 130.2 (d, 3JCF ¼ 8.6 Hz), 129.5,
129.1, 129.0, 128.6, 127.2, 126.5, 122.0, 118.2, 118.1, 115.6 (d,
2JCF ¼ 21.7 Hz), 112.2, 55.0, 42.4. Elemental analysis Found: C, 66.74;
H, 4.14; N, 12.67%; C30H24FN5O2S requires: C, 67.02; H, 4.50; N,
13.03%; LRMS: m/z, 537 (Mþ); C30H24FN5O2S requires: m/z, 537.
4.2.5. 1-(4-Aminosulfonylphenyl)-3-phenyl-
5-[3-(4-methylphenyl)-1-phenylpyrazol-4-yl]-2-pyrazoline (5e)
m.p. 196–198 ꢁC, yield 64%; IR (KBr) 3374 and 3273 (m, N–H
stretch),1593 (s, N–H bend),1337 and 1151 (s, SO2 stretch) cmꢂ1; 1H
4.2.9. 1-(4-Aminosulfonylphenyl)-3-(4-fluorophenyl)-
5-[3-(4-fluorophenyl)-1-phenylpyrazol-4-yl]-2-pyrazoline (5i)
m.p. 212–214 ꢁC, yield 58%; IR (KBr) 3371 and 3259 (m, N–H
NMR (DMSO-d6, 300 MHz):
d
8.31 (s,1H, C50-H pyrazole), 7.82 (d, 2H,
stretch), 1598 (s, N–H bend), 1337 and 1155 (s, SO2 stretch) cmꢂ1
;
J¼ 8.4 Hz, Ar), 7.79–7.76(m, 2H, Ar), 7.66(d, 2H, J¼ 8.1 Hz, Ar), 7.57 (d,
2H, J ¼ 9.0 Hz, Ar), 7.47–7.23 (m, 8H, Ar), 7.03 (d, 2H, J ¼ 8.7 Hz, Ar),
6.99 (s, 2H, ex, SO2NH2), 5.67 (dd, 1H, J ¼ 6.3 Hz, 12.0 Hz, C5-H pyr-
azoline), 4.08 (dd, 1H, J ¼ 12.0 Hz, 17.4 Hz, C4-H pyrazoline), 3.41–
3.34 (m, 1H, C4-H pyrazoline merged with HOD peak), 2.37 (s, 3H,
1H NMR (CDCl3/DMSO-d6, 300 MHz): 7.80 (s, 1H, C50-H pyrazole),
d
7.70–7.66 (m, 4H, Ar), 7.62–7.58 (m, 4H, Ar), 7.36–7.31 (m, 2H, Ar),
7.20–7.17 (m, 3H, Ar), 7.13 (d, 2H, J ¼ 8.7 Hz, Ar), 7.07 (s, 2H, ex,
SO2NH2), 7.03 (d, 2H, J ¼ 8.7 Hz, Ar), 6.97 (d, 2H, J ¼ 9.0 Hz, Ar), 5.13
(dd, 1H, J ¼ 5.7 Hz, 12.0 Hz, C5-H pyrazoline), 3.50 (dd, 1H,
J ¼ 12.0 Hz, 17.1 Hz, C4-H pyrazoline), 2.92–2.82 (m, 1H, C4-H pyr-
azoline merged with HOD peak); 13C NMR (DMSO-d6, 75.5 MHz):
CH3); 13C NMR (DMSO-d6, 75.5 MHz):
d 149.8, 149.7, 146.0, 139.1,
137.7,133.2,131.9,129.7,129.5,129.3,129.2,128.6,127.1,126.3,122.0,
121.7, 118.2, 112.2, 55.1, 42.5, 20.8. Elemental analysis Found: C,
69.41; H, 4.89; N,12.75%; C31H27N5O2S requires: C, 69.77; H, 5.10; N,
13.12%; LRMS: m/z, 533 (Mþ); C31H27N5O2S requires: m/z, 533.
d
162.3 (d, 1JCF ¼ 246 Hz), 162.0 (d, 1JCF ¼ 245 Hz), 149.0,148.3, 146.2,
130.2, (d, 3JCF ¼ 8.4 Hz), 130.1 (d, 3JCF ¼ 8.3 Hz), 129.5, 129.2, 129.0,
2
127.2, 126.4, 122.1, 118.2, 115.7, (d, JCF ¼ 22.3 Hz), 115.6 (d,
2JCF ¼ 22.3 Hz), 112.1, 55.4, 42.8. Elemental analysis Found: C, 65.17;
H, 4.34; N, 12.69%; C30H23F2N5O2S requires: C, 64.85; H, 4.17; N,
12.61%; LRMS: m/z, 555 (Mþ); C30H23F2N5O2S requires: m/z, 555.
4.2.6. 1-(4-Aminosulfonylphenyl)-3-(4-methylphenyl)-
5-[3-(4-methylphenyl)-1-phenylpyrazol-4-yl]-2-pyrazoline (5f)
m.p. 206–208 ꢁC, yield 55%; IR (KBr) 3372 and 3260 (m, N–H
stretch),1597 (s, N–H bend),1337 and 1157 (s, SO2 stretch) cmꢂ1; 1H
4.2.10. 1-(4-Aminosulfonylphenyl)-3-phenyl-
NMR (DMSO-d6, 300 MHz):
d
8.29 (s, 1H, C50-H pyrazole), 7.82 (d,
5-[3-(4-bromophenyl)-1-phenylpyrazol-4-yl]-2-pyrazoline (5j)
m.p. 224–226 ꢁC, yield 58%; IR (KBr) 3372 and 3269 (m, N–H
stretch),1592 (s, N–H bend),1336 and 1151 (s, SO2 stretch) cmꢂ1; 1H
2H, J ¼ 7.8 Hz, Ar), 7.67 (d, 4H, J ¼ 8.1 Hz, Ar), 7.57 (d, 2H, J ¼ 9.0 Hz,
Ar), 7.45–7.40 (m, 2H, Ar), 7.32 (d, 2H, J ¼ 7.8 Hz, Ar), 7.28–7.24 (m,
3H, Ar), 7.02 (d, 2H, J ¼ 9.0 Hz, Ar), 6.98 (s, 2H, ex, SO2NH2), 5.63 (dd,
1H, J ¼ 6.6 Hz, 12.0 Hz, C5-H pyrazoline), 4.05 (dd, 1H, J ¼ 12.0 Hz,
17.1 Hz, C4-H pyrazoline), 3.40–3.26 (m, 1H, C4-H pyrazoline
merged with HOD peak), 2.38 (s, 3H, CH3), 2.34 (s, 3H, CH3); 13C
NMR (DMSO-d6, 300 MHz):
d
8.34 (s, 1H, C50-H pyrazole), 7.82 (d,
2H, J ¼ 7.8 Hz, Ar), 7.76 (d, 2H, J ¼ 8.4 Hz, Ar), 7.72 (d, 4H, J ¼ 8.7 Hz,
Ar), 7.59 (d, 2H, J ¼ 8.7 Hz, Ar), 7.46–7.40 (m, 6H, Ar), 7.05 (d, 2H,
J ¼ 9.0 Hz, Ar), 7.00 (s, 2H, ex, SO2NH2), 5.70 (dd, 1H, J ¼ 6.3 Hz,
12.0 Hz, C5-H pyrazoline), 4.07 (dd, 1H, J ¼ 12.0 Hz, 17.4 Hz, C4-H
pyrazoline), 3.40–3.31 (m, 1H, C4-H pyrazoline merged with HOD
NMR (DMSO-d6, 75.5 MHz): d 149.8, 149.8, 146.1, 139.1, 138.9, 137.7,
133.7, 129.7, 129.4, 129.3, 129.2, 129.1, 127.9, 127.1, 126.3, 122.0, 118.2,
112.1, 55.1, 42.6, 20.9, 20.8. Elemental analysis Found: C, 70.43; H,
5.45; N, 13.01%; C32H29N5O2S requires: C, 70.18; H, 5.34; N, 12.79%;
LRMS: m/z, 547 (Mþ); C32H29N5O2S requires: m/z, 547.
peak); 13C NMR (DMSO-d6, 75.5 MHz):
d 149.8, 148.6, 146.1,
139.1133.3, 131.8, 131.7, 130.0, 129.5, 129.3, 128.7, 127.4, 126.1, 122.2,
121.7, 118.3, 112.3, 55.0, 42.5. Elemental analysis Found: C, 60.02; H,
3.71; N, 11.32%; C30H24BrN5O2S requires: C, 60.20; H, 4.04; N,
11.70%; LRMS: m/z, 597, 599 (Mþ); C30H24BrN5O2S requires: m/z,
597, 599.
4.2.7. 1-(4-Aminosulfonylphenyl)-3-(4-fluorophenyl)-
5-[3-(4-methylphenyl)-1-phenylpyrazol-4-yl]-2-pyrazoline (5g)
m.p. 196–200 ꢁC, yield 65%; IR (KBr) 3372 and 3273 (m, N–H
stretch), 1593 (s, N–H bend), 1337 and 1153 (s, SO2 stretch) cmꢂ1
;
4.2.11. 1-(4-Aminosulfonylphenyl)-3-(4-fluorophenyl)-
1H NMR (DMSO-d6, 300 MHz):
d
8.29 (s,1H, C50-H pyrazole), 7.79 (d,
5-[3-(4-bromophenyl)-1-phenylpyrazol-4-yl]-2-pyrazoline (5k)
m.p. 230–234 ꢁC, yield 65%; IR (KBr) 3373 and 3261 (m, N–H
4H, J ¼ 7.8 Hz, Ar), 7.68 (d, 2H, J ¼ 8.7 Hz, Ar), 7.62 (d, 2H, J ¼ 7.8 Hz,
Ar), 7.40 (t, 2H, J ¼ 7.5 Hz, Ar), 7.25 (m, 5H, Ar), 7.01 (d, 2H, J ¼ 9.0 Hz,
Ar), 6.96 (s, 2H, ex, SO2NH2), 5.63 (dd, 1H, J ¼ 6.0 Hz, 12.0 Hz, C5-H
pyrazoline), 4.02 (dd, 1H, J ¼ 12.0 Hz, 17.1 Hz, C4-H pyrazoline),
3.43–3.32 (m, 1H, C4-H pyrazoline merged with HOD peak), 2.34 (s,
stretch), 1595 (s, N–H bend), 1337 and 1153 (s, SO2 stretch) cmꢂ1
;
1H NMR (DMSO-d6, 300 MHz):
d
8.35 (s, 1H, C50-H pyrazole), 7.81–
7.65 (m, 8H, Ar), 7.59 (d, 2H, J ¼ 9.0 Hz, Ar), 7.47–7.41 (m, 2H, Ar),
7.31–7.25 (m, 3H, Ar), 7.05 (d, 2H, J ¼ 9.0 Hz, Ar), 7.00 (s, 2H, ex,
SO2NH2), 5.70 (dd, 1H, J ¼ 6.3 Hz, 12.3 Hz, C5-H pyrazoline), 4.07
(dd, 1H, J ¼ 12.3 Hz, 18.0 Hz, C4-H pyrazoline), 3.43–3.32 (m, 1H, C4-
H pyrazoline merged with HOD peak); 13C NMR (DMSO-d6,
3H, CH3); 13C NMR (DMSO-d6, 75.5 MHz):
d
161.4 (d, 1JCF ¼ 245 Hz),
149.9, 148.9, 146.0, 139.1, 137.7, 129.8, 129.7, 129.4 (d, 3JCF ¼ 11.1 Hz),
128.6, 128.3, 127.1, 126.3, 122.0, 118.2, 118.1, 115.7 (d, 2JCF ¼ 22.1 Hz),
114.1, 55.2, 42.6, 21.9. Elemental analysis Found: C, 67.78; H, 4.99; N,
13.08%; C31H26FN5O2S requires: C, 67.50; H, 4.75; N, 12.70%; LRMS:
m/z, 551 (Mþ); C31H26FN5O2S requires: m/z, 551.
75.5 MHz);
d
162.7 (d, 1JCF ¼ 247 Hz), 149.0, 148.7, 146.1, 139.0, 133.3,
131.7 (d, 3JCF ¼ 9.5 Hz), 130.1, 129.6, 129.5, 128.2, 126.5, 122.2, 121.7,
118.3, 115.7 (d, 2JCF ¼ 22.1 Hz), 112.3, 55.2, 42.5. Elemental analysis

P.K. Sharma et al. / European Journal of Medicinal Chemistry 45 (2010) 2650–2655
2655
Found: C, 58.12; H, 3.47; N, 11.72%; C30H23BrFN5O2S requires: C,
58.45; H, 3.76; N, 11.36%; LRMS: m/z, 615, 617 (Mþ);
C30H23BrFN5O2S requires: m/z, 615, 617.
a 100 mL volume with concentration of 4 mg/mL of each compound
reconstituted in the dimethylsulphoxide (DMSO). All the plates
were incubated at 37 ꢁC for 24 h. Antimicrobial activity of 11
synthetic compounds was evaluated by measuring the zone of
growth inhibition against the test organisms with zone reader
(HiAntibiotic zone scale). The medium with DMSO as solvent was
used as a negative control whereas media with Ciprofloxacin was
used as positive control. The experiments were performed in trip-
licates. The antimicrobial activities of the compounds were
compared with Ciprofloxacin as standard. MIC of newly synthe-
sized compounds against tested bacteria was determined using
a microplate dilution method [16,17].
4.3. Pharmacological assay
4.3.1. Carrageenan-induced rat paw edema assay
Male Wister albino rats weighing 200–250 g were used
throughout the study. They were kept in the animal house under
standard conditions of light and temperature with free access to
food and water. Food was withdrawn 12 h before and during
experimental hours. The animals were randomly divided into
groups each consisting of six rats. One group of six rats was kept as
control and received tween 80 (95:5). Another group received the
standard drug nimesulide at a dose of 4 mg/kg body weight, i.p.
Other groups of rats were administered the test compounds at
a dose of 50 mg/kg body weight orally. A mark was made on the left
hind paw just beyond the tibiotarsal articulation, so that every time
the paw was dipped up to fixed mark and constant paw volume was
ensured. Paw volumes were measured using a plethysmometer
(model 7140, Ugo Basile, Italy). Thirty minutes after administration
of test and standard drugs, 0.1 mL of 1% w/v of carrageenan
suspension in normal saline was injected into subplanter region of
the left hind paw of all the animals. The initial paw volume was
measured within 30 s of the injection and remeasured again 30, 60,
90, and 120 min after administration of carrageenan. The edema
was expressed as an increase in the volume of paw, and the
percentage of edema inhibition for each rat and each group was
obtained as follows:
Acknowledgements
One of the authors (P.K.) is grateful to the Council of Scientific
and Industrial Research (CSIR), New Delhi for the award of junior
research fellowship. Defence Research and Development Organi-
zation (DRDO), New Delhi is thankfully acknowledged for financial
support in the form of a research project. The authors are thankful
to RSIC, Chandigarh for spectral and analytical data.
References
[1] G. Mu¨ller, Drug Discov. Today 8 (2003) 681–691.
[2] H. Kubinyi, G. Mu¨ller, Chemogenomics in Drug Discovery:
a Medicinal
Chemistry Perspective. Wiley-VCH Verlag GMBH & Co. KGaA, 2004, p. 8, 23.
[3] F.F. Barsoum, H.M. Hosnib, A.S. Girgis, Bioorg. Med. Chem. 14 (2006) 3929–
3937.
[4] P. Rani, V.K. Srivastava, A. Kumar, Eur. J. Med. Chem. 39 (2004) 449–452.
[5] E. Bansal, V.K. Srivastava, A. Kumar, Eur. J. Med. Chem. 36 (2001) 81–92.
¨
[6] G.T. Zitouni, A.O zdemir, Z.A. Kaplancikli, P. Chevallet, Y. Tunali, Phosphorus
% inhibition ¼ ðVt ꢂ V0Þcontrol
ꢂ ðVt ꢂ V0Þtested compound=ðVt ꢂ V0Þcontrol
ꢃ 100
Sulfur Silicon Relat. Elem. 180 (2005) 2717–2724.
[7] M.S. Karthikeyan, B.S. Holla, N.S. Kumari, Eur. J. Med. Chem. 42 (2007) 30–36.
[8] G.T. Zitouni, A. Ozdemir, K. Guven, Arch. Pharm. 338 (2005) 96–104.
[9] G.T. Zitouni, P. Chevallet, F.S. Kiliç, K. Erol, Eur. J. Med. Chem. 35 (2000) 635–641.
[10] A.A. Bekhit, T.A. Aziem, Bioorg. Med. Chem. 12 (2004) 1935–1945.
[11] A.A. Bekhit, H.M.A. Ashour, A.A. Guemei, Arch. Pharm. Chem. Life Sci. 338
(2005) 167–174.
[12] A.A. Bekhit, H.M.A. Rahman, A.A. Guemei, Arch. Phar. Chem. Life Sci. 339
(2006) 81–87.
[13] A.A. Bekhit, H.T.Y. Fahmi, S.A.F. Rostom, A.M. Baraka, Eur. J. Med. Chem. 38
(2003) 27–36.
where Vt ¼ volume of edema at specific time interval and
V0 ¼ volume of edema at zero time interval.
4.3.2. In vitro antimicrobial assay
[14] M.A. Kira, M.O. Abdel-Rahman, K.Z. Gadalla, Tetrahedron Lett. (1969) 109.
[15] M. Bernard, E. Hulley, J. Molenda, K. Stochla, U. Wrzeciono, Pharmazie 41
(1986) 560.
The antibacterial activity of newly synthesized compounds was
evaluated in vitro by agar well diffusion method [21]. All the
microbial cultures were adjusted to 0.5 McFarland standard, which
is visually comparable to a microbial suspension of approximately
1.5 ꢃ 108 CFU/mL [22,23]. 20 mL of Mueller Hinton agar media was
[16] R. Soliman, J. Med. Chem. 22 (1979) 321–325.
[17] C.A. Winter, E.A. Risley, G.W. Nuss, Proc. Soc. Exp. Biol. Med.111 (1962) 544–547.
[18] P.P. Okusa, M. Penge, M. Devleeschouwer, P. Duez, J. Ethnopharmacol. 112
(2007) 476–481.
[19] S.A. Burt, R.D. Reinders, Lett. Appl. Microbiol. 36 (2003) 162–167.
[20] M.K. Bratenko, V.A. Chornoust, M.V. Vovk, Russ. J. Org. Chem. 37 (2001) 556–559.
[21] I. Ahmed, A.J. Beg, J. Ethnopharmacol. 74 (2001) 113–123.
[22] J. McFaraland, J. Am. Med. Assoc. 14 (1907) 1176–1178.
[23] M.J. Andrews, Antimicrob. Chemother. 48 (2001) 5–16.
poured into each Petri plate and plates were swabbed with 100 mL
inocula of the test microorganisms and kept for 15 min for
adsorption. Using sterile cork borer of 8 mm diameter, wells were
bored into the seeded agar plates and these were loaded with