Synthesis and pharmacological evaluation of pyrazole derivatives containing sulfonamide moiety

Article abstract of DOI:10.1007/s00706-016-1694-x

A new series of N-[4-[N-[4-[5-[4-(benzyloxy)phenyl]-1-(substituted phenyl)-1H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acetamide derivatives were synthesized and elucidated by spectral data. All the compounds were subjected to in vitro evaluation for anti-inflammatory (BSA anti-denaturation assay), antioxidant (DPPH radical scavenging assay) and in vivo screening for anti-inflammatory (carrageenan induced rat paw edema inhibition)?activities. Selected active compounds were evaluated for ulcerogenic, lipid peroxidation, and LPS induced TNF-α production inhibition potential. The most active compound in the series showed an in vivo anti-inflammatory efficacy of 83.1?% when compared to diclofenac sodium (81.6?%). Evaluation of ulcer index and biochemical estimation for oxidative stress also revealed that this compound was safe on gastric mucosa and did not induce oxidative stress in tissues. When further tested for LPS induced TNF-α production inhibition in mice, it showed a better inhibition (ID₅₀?=?6.23?mg/kg) when compared to standard inhibitor, SB 203580 (ID₅₀?=?28.40?mg/kg). The p38α MAP kinase docking score of this active compound was also found to be better than that of SB 203580. Graphical abstract: [Figure not available: see fulltext.]

Full text of DOI:10.1007/s00706-016-1694-x

Monatsh Chem
DOI 10.1007/s00706-016-1694-x
ORIGINAL PAPER
Synthesis and pharmacological evaluation of pyrazole derivatives
containing sulfonamide moiety
1
1
2
2
Kanagasabai Somakala
•
Mohammad Amir
•
Vidushi Sharma
•
Sharad Wakode
Received: 30 October 2015 / Accepted: 7 February 2016
Ó Springer-Verlag Wien 2016
Abstract A new series of N-[4-[N-[4-[5-[4-(benzyloxy)-
phenyl]-1-(substituted phenyl)-1H-pyrazol-3-yl]phenyl]
Graphical abstract
H
sulfamoyl]phenyl]acetamide derivatives were synthesized
and elucidated by spectral data. All the compounds were
subjected to in vitro evaluation for anti-inflammatory (BSA
anti-denaturation assay), antioxidant (DPPH radical scav-
enging assay) and in vivo screening for anti-inflammatory
N
H
N
H3C
S
O
O
O
N
N
O
(
carrageenan induced rat paw edema inhibition) activities.
R
Selected active compounds were evaluated for ulcerogenic,
lipid peroxidation, and LPS induced TNF-a production
inhibition potential. The most active compound in the
series showed an in vivo anti-inflammatory efficacy of
Keywords Pyrazole Á TNF-a inhibition Á Docking Á
Anti-inflammatory Á Ulcerogenicity
8
3.1 % when compared to diclofenac sodium (81.6 %).
Evaluation of ulcer index and biochemical estimation for
oxidative stress also revealed that this compound was safe
on gastric mucosa and did not induce oxidative stress in
tissues. When further tested for LPS induced TNF-a pro-
duction inhibition in mice, it showed a better inhibition
Introduction
Non-steroidal anti-inflammatory drugs (NSAIDs) are
therapeutically important class of compounds widely used
in the treatment of various inflammatory diseases particu-
larly arthritic pain [1]. Chronic NSAID therapy effectively
reduces the symptoms of many painful arthritic syndromes,
but invites adverse gastrointestinal (GI) complications
ranging from stomach irritation to life threatening GI
ulceration and bleeding [2]. NSAIDs exert their anti-in-
flammatory effect mainly through inhibition of
cyclooxygenases (COXs), key enzymes in the pros-
taglandin (PG) biosynthesis from arachidonic acid. There
are at least two COX isoforms, COX-1 and COX-2. Con-
stitutive COX-1 is responsible for providing cytoprotection
in GI tract, whereas inducible COX-2 mediates inflam-
mation [3]. Traditional NSAIDs such as aspirin, diclofenac,
ibuprofen, and naproxen are non-selective; however, they
show greater selectivity for COX-1 than COX-2 [4].
Consequently, their long-term clinical use is associated
(
ID50 = 6.23 mg/kg) when compared to standard inhibitor,
SB 203580 (ID50 = 28.40 mg/kg). The p38a MAP kinase
docking score of this active compound was also found to be
better than that of SB 203580.
Electronic supplementary material The online version of this
article (doi:10.1007/s00706-016-1694-x) contains supplementary
material, which is available to authorized users.
&
Mohammad Amir
mamir_s2003@yahoo.co.in
1
2
Department of Pharmaceutical Chemistry, Faculty of
Pharmacy, Hamdard University, New Delhi 110062, India
Department of Pharmaceutical Chemistry, University of
Delhi (Delhi Institute of Pharmaceutical Sciences and
Research), New Delhi 110017, India
123

K. Somakala et al.
with GI toxicity [5]. Thus the discovery of COX-2 pro-
vided the rationale for the development of drugs devoid of
GI toxicity while retaining clinical efficacy as anti-in-
flammatory agents. But the recent report showed that
selective COX-2 inhibitors could lead to adverse cardio-
vascular side effects [6]. Therefore development of novel
compounds having anti-inflammatory activity with an
improved safety profile is still a necessity.
presence of anhydrous K CO in dimethyl formamide
2
3
(DMF) at room temperature. Treatment of 1 with
4-aminoacetophenone in presence of aqueous NaOH in
methanol at room temperature yielded 1-(4-aminophenyl)-
3-[4-(benzyloxy)phenyl]prop-2-en-1-one (2). This chal-
cone was then refluxed with different aryl hydrazines in
ethanol to give 4-[5-[4-(benzyloxy)phenyl]-1-(substituted
phenyl)-1H-pyrazol-3-yl]anilines 3a–3o. Synthesis of tar-
get compounds 4a–4o was achieved by stirring pyrazoles
3a–3o with p-acetamidobenzenesulfonyl chloride in the
presence of tetrahydrofuran (THF) and pyridine. Synthesis
of these compounds is outlined in Scheme 1.
p38 mitogen activated protein (MAP) kinase is a
member of the intracellular family of MAP kinases
implicated in the phosphorylation cascade leading to the
release of tumour necrosis factor-alpha (TNF-a) and other
cytokines [7]. TNF-a is a multifunctional cytokine which
participates in the pathogenesis of various diseases
including autoimmune, inflammatory, and neurodegenera-
tive diseases. TNF-a also mediates many of the
inflammatory processes in rheumatoid arthritis (RA)
including immune cell activation, proliferation, apoptosis
and regulation of leukocyte movement [8, 9]. Therefore
TNF-a has emerged as an attractive target to reduce the
inflammatory conditions.
1
H NMR spectra of titled compounds 4a–4o showed a
singlet for CH CO and CONH protons at d = 2.06–3.01
3
and at 8.32–8.90 ppm, respectively. The singlets of SO NH
2
and OCH protons were observed at d = 10.18–10.31 and
2
1
3
5.15–5.22 ppm, respectively. In the C NMR spectra of
compounds 4a–4o, CH3 and OCH2 carbon peaks were
observed at d = 39.49–23.78 and 72.26–70.24 ppm,
respectively. Furthermore, CONH carbon peaks were
observed downfield at d = 172.47–169.92 ppm. Mass
spectra of all the compounds showed a molecular ion peak
Pyrazoles and their derivatives are widely known for their
excellent effectiveness as analgesics and antipyretics. Anti-
pyrin was the first pyrazoline derivative used in the
management of pain and inflammation. Phenylbutazone and
its derivative, oxyphenbutazone, were potent anti-inflam-
matory agents. The marketed selective COX-2 inhibitor
?
M at an m/z corresponding to their molecular formula.
In vitro anti-inflammatory activity
The synthesized compounds 4a–4o were screened for anti-
inflammatory activity by inhibition of albumin denatura-
tion technique. Compound 4a showed the maximum anti-
inflammatory activity of 89 % when compared to the
standard diclofenac sodium (83.6 %). Compounds 4c
(87.5 %) and 4b (84.5 %) also exhibited higher anti-in-
flammatory activity in comparison to diclofenac sodium.
Other compounds were found to have anti-inflammatory
activity in the range of 62.9–82.2 % (Table 1). From the
results it can be stated that synthesized analogs are capable
of controlling the production of auto antigens due to pre-
vention of denaturation of proteins and the findings
suggested that these compounds may show good in vivo
anti-inflammatory activity.
(
celecoxib) having a pyrazole moiety also showed high anti-
inflammatory activity with reduced GI toxicity [10]. Besides
these, many pyrazole derivatives are also reported in litera-
ture having potent anti-inflammatory activity [11–15].
Furthermore, sulfonamide derivatives are well known
pharmaceutical agents since this group has been a part of
most of the drug structures due to stability and tolerance in
human beings [16]. This group is also the key constituent of
COX inhibitors such as celecoxib [17] and valdecoxib [18].
Encouraged by these observations and in continuation of
our research program on the synthesis of 5-membered hete-
rocyclic compounds as anti-inflammatory agents [19–22], it
was attempted in the present study to synthesize and evaluate
anti-inflammatory activity and TNF-a inhibitory properties
of some new pyrazole derivatives bearing sulfonamide
pharmacophore. The most potent derivative was also docked
into the active site of p38 MAPK so as to explore the inter-
action mechanism between the inhibitor and the receptor.
Antioxidant activity
The free radical scavenging activity of the compounds 4a–
4o was evaluated by DPPH colorimetric method at 1 mM
concentration. The degree of change in absorbance with
respect to control was calculated as antioxidant potential.
Compounds 4a (76.0 %), 4c (74.2 %), and 4b (74 %)
showed antioxidant activity within the same range in
comparison to the standard, butylated hydroxyl anisole
Results and discussion
Chemistry
(BHA) (73 %). All the other compounds also exhibited
4-(Benzyloxy)benzaldehyde (1) was prepared by treating
4-hydroxybenzaldehyde and benzyl chloride in the
good antioxidant activity which is in the range of
71–40.4 % (Table 1). The study revealed that electron
1
23

Synthesis and pharmacological evaluation of pyrazole derivatives containing sulfonamide moiety
Scheme 1
O
O
Anhy. K CO3
H
2
H
DMF
O
OH
1
Cl
4
-Aminoacetophenone,
H N
2
aq. NaOH, CH OH
3
O
Aryl hydrazines,
C H OH
2
5
N
2
H N
2
O
3
a-3o
N
O
R
R = a: 2-chloro, b: 3-chloro, c: 4-chloro, d: 3,4-
dichloro, e: 4-fluoro , f: 4-fluoro-3-chloro, g: 4-
bromo, h: 4-nitro, i: 2,4-dinitro, j: 2-methyl, k: 3-
methyl, l: 4-methyl, m: 2,6-dimethyl, n: 2-
methoxy, o: 4-methoxy
p-Acetamidobenzenesulfonyl
chloride, pyridine, THF
H
N
H C
3
HN
S
O
O
O
4a-4o
N
N
O
R
withdrawing groups increased the antioxidant potential
which may be due to intensification of positive charge
associated with negative inductive effect of these groups.
The positive charge intensification may lead to free radical
quenching. Moreover, electron withdrawing groups are
themselves good free radical quenchers. On the other hand,
substitution with electron releasing groups was found to
decrease the radical scavenging potential possibly due to
their positive inductive effect.
Compound 4a having the 2-chlorophenyl group attached
to position 1 of the pyrazole ring emerged as the most
promising compound of the series showing anti-inflam-
matory activity (83.1 %) slightly higher than diclofenac
sodium, whereas compound 4c and 4b having 4-chloro-
and 3-chlorophenyl groups showed anti-inflammatory
activity 82.1 and 80.5 % respectively which is within the
range of standard drug diclofenac sodium. When the
chlorophenyl group was replaced by a 4-fluorophenyl
(
4e), 4-bromophenyl (4g), or 4-nitrophenyl (4h) group,
the activity was found to be decreased (74.0, 72.6, and
2.1 %, respectively). A further decrease in activity was
In vivo anti-inflammatory activity
7
The anti-inflammatory activity of the synthesized com-
pounds 4a–4o was evaluated by carrageenan induced paw
edema method of Winter et al. The compounds were
tested at equimolar oral dose relative to 10 mg/kg
diclofenac sodium. The tested compounds showed anti-
inflammatory activity ranging from 52.3 to 83.1 %,
whereas standard drug diclofenac sodium showed 81.7 %.
observed for compounds 4d (71.4 %) and 4f (70.7 %)
which possessed two electron withdrawing groups i.e. 3,4-
dichlorophenyl and 4-fluoro-3-chlorophenyl, respectively.
Compounds 4i–4o showed moderate to low activity
ranging from 52.3 to 66.1 % which possessed -2,4-dinitro,
–CH , and –OCH groups attached to position 1 of the
3
3
pyrazole ring (Table 2).
123

K. Somakala et al.
Table 1 In vitro anti-inflammatory activity (BSA denaturation inhibition assay) and % DPPH activity
a
b
Compounds
BSA denaturation inhibition assay
% DPPH activity (at 1 mM)
Mean absorbance
% Inhibition of denaturation ± SD
Mean absorbance
% Antioxidant activity ± SD
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
a
b
c
d
e
f
0.353
0.345
0.351
0.333
0.341
0.325
0.337
0.334
0.311
0.307
0.315
0.319
0.305
0.305
0.318
0.343
89.0 ± 2.53
84.5 ± 2.83
87.5 ± 1.88
78.1 ± 1.85
82.2 ± 4.32
73.8 ± 3.51
80.0 ± 2.53
78.8 ± 2.16
66.1 ± 0.82
64.4 ± 0.31
68.3 ± 2.64
70.8 ± 4.85
63.3 ± 2.74
62.9 ± 4.97
70.1 ± 3.25
83.6 ± 2.03
0.197
0.214
0.207
0.282
0.239
0.286
0.253
0.26
76.0 ± 0.64
74.0 ± 0.56
74.8 ± 0.49
65.7 ± 0.93
71.0 ± 0.8
65.3 ± 0.98
69.2 ± 0.56
68.3 ± 0.49
54.7 ± 0.51
53.6 ± 0.81
59.7 ± 0.73
61.8 ± 0.61
52.1 ± 0.98
40.4 ± 0.61
60.3 ± 0.73
73.0 ± 1.27
g
h
i
0.372
0.381
0.331
0.314
0.394
0.49
j
k
l
m
n
o
0.326
0.222
Standard
SD standard deviation (average of three determinations)
a
Diclofenac sodium
b
Butylated hydroxyl anisole
Ulcerogenicity
lipid peroxidation (6.53 ± 0.22) whereas the control group
displayed a lipid peroxidation of 3.23 ± 0.10 (Table 3).
Thus this study showed that the synthesized compounds
have inhibited the induction of gastric mucosal lesions and
the results further suggested that their protective action
might be related to the inhibition of lipid peroxidation in
the gastric mucosa.
Compounds 4a–4c, 4e, and 4g exhibiting significant anti-
inflammatory activity were further screened for their
ulcerogenic potential. It was observed that the tested
compounds exhibited a better GI safety profile, severity
index ranging from 0.42 ± 0.38 to 0.92 ± 0.66, whereas
standard drug diclofenac sodium showed a high severity
index of 1.83 ± 0.25. Most potent compounds 4a and 4c
showing high anti-inflammatory activity showed a reduc-
tion in severity index (0.42 ± 0.38 and 0.67 ± 0.52,
respectively) in comparison to standard drug (Table 3).
From the results obtained it may be suggested that due to
decrease in the ulcerogenic activity the compounds may
cause less gastric ulceration and disruption of gastric
epithelial cells.
LPS induced TNF-a production inhibition
Subsequently we evaluated the inhibitory activity of the
derivatives 4a–4c, 4e, and 4g in the LPS induced TNF-a
production in mice. Compound 4a showed the most
effective activity (ID50 = 6.23 mg/kg), which was more
than the standard compound, SB 203580 (ID50 = 28.40 -
mg/kg). Analogs 4c and 4b also demonstrated good TNF-a
inhibitory efficacy with ID50 values of 10.96 and 10.83 mg/
kg, respectively. Compounds 4e (52.8 %) and 4g (52.8 %)
showed activity more or less similar to that of the standard
inhibitor, SB 203580 which showed 52.1 % inhibition
(Table 3).
Lipid peroxidation
The compounds screened for ulcerogenic activity were also
analyzed for lipid peroxidation. Lipid peroxidation is
measured as nmol of MDA per 100 mg of gastric mucosa
tissue. The tested compounds 4a–4c, 4e, and 4g exhibited
significant reduction in lipid peroxidation (in the range of
Molecular docking study
4
.69 ± 0.87–6.12 ± 0.50). Most active anti-inflammatory
The glide score value of SB 203580 was found to be -9.163,
while that of compound 4a was -10.058, indicating the
strong interaction of the molecule with the enzyme
(Table 2). The best binding mode of SB 203580 (p38a
compounds 4a and 4c showed reduction in lipid peroxi-
dation (4.69 ± 0.87 and 5.26 ± 1.50, respectively). The
standard drug, diclofenac sodium showed the maximum
1
23

Synthesis and pharmacological evaluation of pyrazole derivatives containing sulfonamide moiety
Table 2 Anti-inflammatory activity and XP docking glide score of compounds 4a–4o and standards
3
Increase in paw edema/cm ± SEM
d
Compounds
% Inhibition
Activity relative to diclofenac
Glide score
a
4
4
4
4
4
4
4
4
4
4
4
4
4
4
4
a
b
c
d
e
f
0.29 ± 0.08
83.1
80.5
82.1
71.4
74.0
70.7
72.6
72.1
56.4
54.1
60.7
66.1
53.3
52.3
65.5
–
102.1
98.9
100.5
87.7
90.9
86.8
89.2
88.6
69.3
66.4
74.5
81.2
65.4
64.2
80.5
–
-10.058
-10.107
-9.678
-11.324
-10.083
-10.581
-8.706
-8.240
-10.211
-11.192
-11.124
-9.320
-10.202
-10.636
-6.038
–
c
0.33 ± 0.14
c
0.30 ± 0.06
a
0.48 ± 0.06
a
0.44 ± 0.17
a
0.50 ± 0.15
a
g
h
i
0.46 ± 0.13
b
0.47 ± 0.12
b
0.74 ± 0.17
a
j
0.78 ± 0.24
c
k
l
0.66 ± 0.11
a
0.57 ± 0.23
a
m
n
o
0.79 ± 0.10
a
0.81 ± 0.27
a
0.58 ± 0.13
a
Control
1.69 ± 0.04
a
Diclofenac sodium
SB 203580
0.31 ± 0.01
81.7
–
100
-4.875
-9.163
–
–
Anti-inflammatory activity of the compounds were compared with respect to control. Data were analyzed by unpaired student’s t test for n = 6.
All data are significantly different from control
a
p \ 0.05
b
p \ 0.01
c
p \ 0.001
d
Glide score denotes g score obtained for docking with p38a MAP kinase (PDB ID: 3D83)
Table 3 Ulcerogenic, lipid peroxidation, and LPS induced TNF-a production inhibition potential
Compounds
Mean severity index ± SEM Lipid peroxidation nmol MDA/100 mg tissue % TNF-a inhibition ± SEM ID50/mg/kg
b
b
a
b
a
b
a
a
a
a
a
a
a
b
a
b
b
4
4
4
4
4
a
b
c
e
g
0.42 ± 0.38
0.83 ± 0.26
0.67 ± 0.52
0.83 ± 0.41
0.92 ± 0.66
0.00 ± 0.00
4.69 ± 0.87
5.89 ± 0.44
5.26 ± 1.50
5.69 ± 0.67
6.12 ± 0.50
3.23 ± 0.10
6.53 ± 0.22
–
64.1 ± 1.24
56.4 ± 1.80
57.3 ± 0.66
52.8 ± 1.00
52.8 ± 0.85
–
6.23
10.83
10.96
–
–
Control
–
a
Diclofenac sodium 1.83 ± 0.25
SB203580
–
–
b
–
52.1 ± 1.96
28.40
Relative to standard and data were analyzed by unpaired student’s t test for n = 6
a
p \ 0.01
b
p \ 0.001
prototype inhibitor) and compound 4a in the active site of
p38a MAP kinase are presented in Fig. 1a, b, respectively.
The docked pose of compound 4a showed a hydrogen bond
Conclusion
We have reported here a simple synthesis of a series of new
pyrazole derivatives bearing a sulfonamide pharma-
cophore. The compounds having chloro, fluoro, and bromo
groups in position 1 of the pyrazole ring have exhibited
very good anti-inflammatory activity in comparison to
diclofenac sodium along with good antioxidant potential.
˚
interaction with Met 109 (OÁÁÁHN, 2.13 A) and Thr 106
˚
(
OÁÁÁHN, 2.40 A) (Fig. 2). Interestingly, compound 4a also
forms a p–p interaction between a phenyl ring and Phe 169.
Standard SB 203580 was found to show two hydrogen bond
interactions with Glu 71 and Asp 168.
123

K. Somakala et al.
Compound 4a showing high anti-inflammatory activity was
found to be safe on gastric mucosa with least ability to
induce oxidative stress in tissues. The LPS induced TNF-a
production inhibition for compound 4a was found to be
superior in comparison to that of SB 203580 in mice.
Furthermore, molecular docking study of compound 4a
revealed favorable orientation within the active binding site
of p38a MAP kinase with a comparatively higher docking
score than SB 203580. Therefore, compound 4a represent a
fruitful matrix for development of anti-inflammatory
agents that would deserve further investigation and
derivatization.
carbonate (7.5 mmol) in 10 cm of DMF. The solution was
then poured into ice cold water and the precipitate thus
3
obtained was filtered, dri ed and crystallized from ethanol.
Yield: 92 %; m.p.: 74–75 °C (Ref. [23] 65–66 °C).
1
-(4-Aminophenyl)-3-[4-(benzyloxy)phenyl]prop-2-en-1-
one (2)
-(Benzyloxy)benzaldehyde (1, 5 mmol) and 4-aminoace-
4
3
tophenone (5 mmol) in the presence of 10 cm aqueous
3
sodium hydroxide (4 %) and 25 cm methanol was stirred
at room temperature for 4h. Yellow crystalline precipitate
was obtained which was filtered, washed with water, and
dried. Yield 89 %, m.p.: 182–183 °C; IR (KBr): vꢀ = 3487,
-
1
1
3209 (NH), 3021 (CH), 1665 (C=O) cm
; H NMR
(
CDCl ): d = 5.09 (s, 2H, OCH ), 5.72 (s, 2H, NH , D O
3
2
2
2
Experimental
exchangeable), 7.05 (d, J = 7.6 Hz, 2H, ArH), 7.42 (d,
J = 7.6 Hz, 2H, ArH), 7.43–7.64 (m, 5H, ArH), 7.67 (d,
J = 8.4 Hz, 2H, ArH), 7.72 (d, J = 15.2 Hz, 1H, CO–
CH=CH), 7.91 (d, J = 15.2 Hz, 1H, CO–CH=CH), 7.97
All reagents and solvents were of laboratory grade and
were procured from Merck (Darmstadt, Germany) and S.D.
Fine chemicals (Delhi, India). Melting points were deter-
mined in open capillaries using Digital Auto Melting Point
Apparatus (Labtronics). Purity of the compounds was
ascertained by TLC using precoated aluminium TLC plates
1
3
(
d, J = 8.4 Hz, 2H, ArH) ppm; C NMR (CDCl3):
d = 70.1 (OCH ), 113.9 (CH=CH), 115.3 (CH=CH),
1
1
2
19.9, 127.5, 128.2, 128.3, 128.6, 128.7, 130.0, 131.0,
36.5, 142.9, 151.2, 160.5 (C=O) ppm.
(
Merck) visualized in a UV/Visible chamber. Infrared
spectra were recorded on Perkin-Elmer 1720 FTIR spec-
General method for the synthesis of 4-[5-[4-(benzyloxy)
phenyl]-1-(substituted phenyl)-1H-pyrazol-3-yl]anilines
3a–3o
1
13
trometer. H NMR spectra (400 MHz) and C NMR
spectra (100 MHz, with complete proton decoupling) were
recorded on a Bruker Avance-400 instrument. Chemical
shifts were reported in ppm downfield from tetramethyl-
silane (TMS) as the internal standard. Mass spectra were
recorded on Jeol SX-102/DA-6000 spectrometer. Elemen-
tal analyses (C, H and N) were undertaken with a CHNS
Vario EL III (Elementar Analysen systeme GmbH, Ger-
many) and the results are within ± 0.4 % of theoretical
values.
Amixture of 1-(4-aminophenyl)-3-[4-(benzyloxy)phenyl]
prop-2-en-1-one (2, 5 mmol) and different aryl hydrazines
3
(7.5 mmol) in 25 cm of ethanol was refluxed at 60 °C for
3–7 h. The solution was cooled at room temperature and
poured into ice cold water. The precipitate thus obtained
was filtered, washed with water, dried, and recrystallized
from ethanol.
4
-[5-[4-(Benzyloxy)phenyl]-1-(2-chlorophenyl)-1H-pyra-
zol-3-yl]aniline (3a, C28 22ClN O)
Yield 88 %; m.p.: 126–127 °C; IR (KBr): vꢀ = 3522, 3351
4
-(Benzyloxy)benzaldehyde(1)
H
3
Equimolar quantities of 4-hydroxybenzaldehyde and ben-
zyl chloride (5 mmol each) were stirred at room
temperature for 8 h in the presence of anhydrous potassium
-
1 1
(NH), 3035 (CH), 1596 (C=N), 736 (C–Cl) cm ; H NMR
(DMSO-d ): d = 5.12 (s, 2H, OCH ), 5.17 (s, 2H, NH
,
2
6
2
Fig. 1 A suggested binding mode of (a) SB 203580 and (b) compound 4a into p38a MAP kinase (PDB code: 3D83)
1
23

Synthesis and pharmacological evaluation of pyrazole derivatives containing sulfonamide moiety
Fig. 2 Docked pose of
compound 4a in the binding site
of p38a MAP kinase showing
hydrogen bond interaction
D O exchangeable), 6.65 (s, 1H, pyrazole H-4), 7.05–7.82
2
(OCH ), 115.7 (pyrazole C ), 118.3, 120.5, 128.4, 128.9,
2 4
1
3
(
m, 17H, Ar–H) ppm; C NMR (DMSO-d ): d = 68.9
129.1, 129.3, 130.1, 130.3, 131.2, 131.6, 132.0, 132.1,
132.4, 132.8, 132.9, 136.1, 143.6, 144.1, 160.5 (pyrazole
C ), 163.5 (pyrazole C ) ppm.
6
(
OCH ), 115.6 (pyrazole C ), 118.7, 120.1, 128.2, 128.9,
2
4
1
1
29.1, 129.3, 130.0, 130.2, 131.1, 131.8, 132.0, 132.2,
32.4, 132.8, 132.9, 136.2, 143.6, 144.0, 160.5 (pyrazole
5
3
4-[5-[4-(Benzyloxy)phenyl]-1-(4-fluorophenyl)-1H-pyra-
C ), 163.4 (pyrazole C ) ppm.
5
3
zol-3-yl]aniline (3e, C H FN O)
2
8
22
3
4
-[5-[4-(Benzyloxy)phenyl]-1-(3-chlorophenyl)-1H-pyra-
Yield 85 %; m.p.: 132–133 °C; IR (KBr): vꢀ = 3565, 3350
(NH), 3032 (CH), 1585 (C=N) cm ; H NMR (DMSO-
-1 1
zol-3-yl]aniline (3b, C H ClN O)
2
8
22
3
Yield 89 %; m.p.: 120 °C; IR (KBr): vꢀ = 3520, 3340
d ): d = 5.18 (s, 2H, OCH ), 5.22 (s, 2H, NH , D O
6
2
2
2
-
1 1
NH), 3035 (CH), 1592 (C=N), 741 (C–Cl) cm ; H NMR
(
exchangeable), 6.66 (s, 1H, pyrazole H-4), 7.05–7.82 (m,
1
3
(
DMSO-d ): d = 5.16 (s, 2H, OCH ), 5.19 (s, 2H, NH ,
17H, Ar–H) ppm;
(OCH ), 115.7 (pyrazole C ), 118.4, 120.2, 128.2, 128.8,
C NMR (DMSO-d ): d = 68.5
6
2
2
6
D O exchangeable), 6.62 (s, 1H, pyrazole H-4), 7.02–7.81
2
2
4
1
3
(
m, 17H, Ar–H) ppm; C NMR (DMSO-d ): d = 68.3
129.2, 129.4, 130.0, 130.3, 131.2, 131.7, 132.2, 132.4,
6
(
OCH ), 115.9 (pyrazole C ), 118.3, 120.8, 128.2, 128.8,
2
132.8, 136.3, 143.5, 144.2, 160.4 (pyrazole C ), 163.3
5
4
1
1
29.1, 129.2, 129.8, 130.4, 131.1, 131.7, 132.1, 132.3,
32.4, 132.7, 132.8, 136.4, 143.5, 144.2, 160.4 (pyrazole
(pyrazole C ) ppm.
3
4
1
-[5-[4-(Benzyloxy)phenyl]-1-(3-chloro-4-fluorophenyl)-
C ), 163.3 (pyrazole C ) ppm.
5
3
H-pyrazol-3-yl]aniline (3f, C H ClFN O)
2
8
21
3
4
-[5-[4-(Benzyloxy)phenyl]-1-(4-chlorophenyl)-1H-pyra-
Yield 88 %; m.p.: 145–146 °C; IR (KBr): vꢀ = 3560, 3354
(NH), 3026 (CH), 1591 (C=N), 729 (C–Cl) cm ; H NMR
-1 1
zol-3-yl]aniline (3c, C H ClN O)
2
8
22
3
Yield 80 %; m.p.: 163 °C; IR (KBr): vꢀ = 3496, 3348
(DMSO-d ): d = 5.16 (s, 2H, OCH ), 5.22 (s, 2H, NH ,
6
2
2
-
1 1
NH), 3029 (CH), 1599 (C=N), 749 (C–Cl) cm ; H NMR
(
D O exchangeable), 6.70 (s, 1H, pyrazole H-4), 7.06–7.84
2
1
3
(
DMSO-d ): d = 5.15 (s, 2H, OCH ), 5.19 (s, 2H, NH ,
(m, 16H, Ar–H) ppm; C NMR (DMSO-d ): d = 68.3
6
2
2
6
D O exchangeable), 6.62 (s, 1H, pyrazole H-4), 7.09–7.83
2
(OCH ), 115.4 (pyrazole C ), 118.6, 120.2, 128.4, 128.9,
2
4
1
3
(
m, 17H, Ar–H) ppm; C NMR (DMSO-d ): d = 68.9
129.2, 129.3, 130.1, 130.3, 131.1, 131.7, 132.0, 132.1,
132.4, 132.7, 132.9, 136.4, 143.5, 144.2, 161.5 (pyrazole
C ), 163.6 (pyrazole C ) ppm.
6
(
OCH ), 115.3 (pyrazole C ), 118.2, 120.4, 128.3, 128.8,
2
4
1
1
29.1, 129.3, 129.8, 130.2, 131.3, 131.8, 132.1, 132.3,
5
3
32.5, 136.2, 143.5, 144.2, 160.6 (pyrazole C ), 163.7
5
4-[5-[4-(Benzyloxy)phenyl]-1-(4-bromophenyl)-1H-pyra-
(
pyrazole C ) ppm.
3
zol-3-yl]aniline (3g, C H BrN O)
2
8
22
3
4
-[5-[4-(Benzyloxy)phenyl]-1-(3,4-dichlorophenyl)-1H-
Yield 87 %; m.p.: 132–133 °C; IR (KBr): vꢀ = 3520, 3368
(NH), 3033 (CH), 1585 (C=N) cm ; H NMR (DMSO-
-1 1
pyrazol-3-yl]aniline (3d, C H Cl N O)
2
8
21
2 3
Yield 89 %; m.p.: 127–129 °C; IR (KBr): vꢀ = 3570, 3345
d ): d = 5.16 (s, 2H, OCH ), 5.17 (s, 2H, NH , D O
6
2
2
2
-
1 1
NH), 3037 (CH), 1596 (C=N), 742 (C–Cl) cm ; H NMR
(
exchangeable), 6.65 (s, 1H, pyrazole H-4), 7.05–7.82 (m,
1
3
(
DMSO-d ): d = 5.13 (s, 2H, OCH ), 5.22 (s, 2H, NH ,
17H, Ar–H) ppm;
(OCH ), 115.8 (pyrazole C ), 118.6, 120.8, 128.3, 128.8,
C NMR (DMSO-d ): d = 68.7
6
2
2
6
D O exchangeable), 6.71 (s, 1H, pyrazole H-4), 7.15–7.82
2
2
4
1
3
(
m, 16H, Ar–H) ppm; C NMR (DMSO-d ): d = 68.6
129.1, 129.2, 129.7, 130.2, 131.3, 131.8, 132.1, 132.3,
6
123

K. Somakala et al.
1
32.8, 136.2, 143.5, 144.2, 160.3 (pyrazole C ), 163.7
5
d ): d = 2.32 (s, 3H, CH ), 5.22 (s, 2H, OCH ), 5.25 (s,
6
3
2
(
pyrazole C ) ppm.
3
2H, NH , D O exchangeable), 6.65 (s, 1H, pyrazole H-4),
2 2
1
3
.05–7.82 (m, 17H, Ar–H) ppm; C NMR (DMSO-d6):
7
4
3
-[5-[4-(Benzyloxy)phenyl]-1-(4-nitrophenyl)-1H-pyrazol-
d = 16.1 (CH ), 68.8 (OCH ), 115.6 (pyrazole C ), 118.5,
3
2
4
-yl]aniline (3h, C H N O )
2
8 22 4 3
1
1
20.8, 128.4, 128.9, 129.0, 129.5, 130.1, 130.3, 131.2,
31.7, 132.3, 132.7, 132.9, 136.1, 143.7, 144.2, 160.5
Yield 88 %; m.p.: 182–183 °C; IR (KBr): vꢀ = 3560, 3341
-
1 1
NH), 3039 (CH), 1597 (C=N) cm ; H NMR (DMSO-
(
(pyrazole C ), 163.7 (pyrazole C ) ppm.
5 3
d ): d = 5.09 (s, 2H, OCH ), 5.17 (s, 2H, NH , D O
6
2
2
2
exchangeable), 6.69 (s, 1H, pyrazole H-4), 7.07–7.81 (m,
3
4-[5-[4-(Benzyloxy)phenyl]-1-(2,6-dimethylphenyl)-1H-
1
1
7H, Ar–H) ppm;
C NMR (DMSO-d ): d = 68.6
pyrazol-3-yl]aniline (3m, C H N O)
30 27 3
6
(
OCH ), 115.6 (pyrazole C ), 118.4, 120.6, 128.3, 128.8,
2
Yield 87 %; m.p.: 132–133 °C; IR (KBr): vꢀ = 3525, 3328
(NH), 3024 (CH), 1590 (C=N) cm ; H NMR (DMSO-
4
-1 1
1
1
29.2, 129.3, 129.8, 130.2, 131.1, 131.6, 132.2, 132.5,
32.9, 136.1, 143.6, 144.1, 161.2 (pyrazole C ), 163.6
5
d ): d = 2.10 (s, 6H, 2 CH ), 5.12 (s, 2H, OCH ), 5.17 (s,
6
3
2
(
pyrazole C ) ppm.
3
2H, NH , D O exchangeable), 6.68 (s, 1H, pyrazole H-4),
2 2
1
.01–7.88 (m, 16H, Ar–H) ppm; C NMR (DMSO-d6):
3
7
4
-[5-[4-(Benzyloxy)phenyl]-1-(2,4-dinitrophenyl)-1H-
d = 15.6 (2 CH ), 68.8 (OCH ), 115.6 (pyrazole C ),
3
2
4
pyrazol-3-yl]aniline (3i, C H N O )
2
8 21 5 5
1
18.3, 120.5, 128.2, 128.8, 129.1, 129.5, 130.1, 130.4,
31.2, 131.8, 132.1, 132.3, 132.6, 132.8, 132.9, 136.3,
43.6, 144.1, 161.7 (pyrazole C ), 163.5 (pyrazole C )
Yield 89 %; m.p.: 148–149 °C; IR (KBr): vꢀ = 3525, 3352
1
-1 1
NH), 3085 (CH), 1584 (C=N) cm ; H NMR (DMSO-
(
1
5
3
d ): d = 5.11 (s, 2H, OCH ), 5.17 (s, 2H, NH , D O
exchangeable), 6.68 (s, 1H, pyrazole H-4), 7.10–7.86 (m,
1
1
6
2
2
2
ppm.
3
6H, Ar–H) ppm;
C NMR (DMSO-d ): d = 68.8
4-[5-[4-(Benzyloxy)phenyl]-1-(2-methoxyphenyl)-1H-pyra-
6
(
OCH ), 115.9 (pyrazole C ), 118.4, 120.8, 128.4, 128.8,
2
zol-3-yl]aniline (3n, C H N O)
29 25 3
4
1
1
29.1, 129.4, 130.0, 130.4, 131.2, 131.8, 132.0, 132.1,
32.5, 132.7, 132.9, 136.3, 143.6, 144.0, 160.2 (pyrazole
Yield 88 %; m.p.: 158–160 °C; IR (KBr): vꢀ = 3520, 3344
(NH), 3019 (CH), 1592 (C=N) cm ; H NMR (DMSO-
-1 1
C ), 163.8 (pyrazole C ) ppm.
5 3
d ): d = 3.52 (s, 3H, OCH ), 5.18 (s, 2H, OCH ), 5.27 (s,
6
3
2
2
H, NH , D O exchangeable), 6.71 (s, 1H, pyrazole H-4),
2 2
4
-[5-[4-(Benzyloxy)phenyl]-1-(2-methylphenyl)-1H-pyra-
1
.09–7.82 (m, 17H, Ar–H) ppm; C NMR (DMSO-d6):
3
7
zol-3-yl]aniline (3j, C H N O)
2
9 25 3
d = 53.2 (OCH ), 68.5 (OCH ), 115.8 (pyrazole C ),
1
3
2
4
Yield 86 %; m.p.: 134–135 °C; IR (KBr): vꢀ = 3562, 3347
18.6, 120.4, 128.3, 128.9, 129.1, 129.3, 129.8, 130.2,
31.1, 131.6, 132.0, 132.2, 132.4, 132.7, 132.8, 136.3,
43.6, 144.1, 160.6 (pyrazole C ), 163.8 (pyrazole C )
-1 1
NH), 3021 (CH), 1585 (C=N) cm ; H NMR (DMSO-
(
1
d ): d = 2.18 (s, 3H, CH ), 5.10 (s, 2H, OCH ), 5.27 (s,
6
3
2
1
5
3
2
7
H, NH , D O exchangeable), 6.61 (s, 1H, pyrazole H-4),
2 2
ppm.
1
.06–7.82 (m, 17H, Ar–H) ppm; C NMR (DMSO-d6):
3
d = 16.3 (CH ), 68.5 (OCH ), 115.8 (pyrazole C ), 118.5,
4-[5-[4-(Benzyloxy)phenyl]-1-(4-methoxyphenyl)-1H-pyra-
3
2
4
1
1
1
20.7, 128.3, 128.8, 129.2, 129.3, 129.8, 130.1, 131.1,
31.7, 132.1, 132.3, 132.6, 132.8, 132.9, 136.2, 143.7,
44.0, 160.4 (pyrazole C ), 163.9 (pyrazole C ) ppm.
zol-3-yl]aniline (3o, C H N O)
2
9 25 3
Yield 88 %; m.p.: 184–185 °C; IR (KBr): vꢀ = 3525, 3353
(NH), 3028 (CH), 1575 (C=N) cm ; H NMR (DMSO-
-
1 1
5
3
d ): d = 3.71 (s, 3H, OCH ), 5.21 (s, 2H, OCH ), 5.25 (s,
6
3
2
4
-[5-[4-(Benzyloxy)phenyl]-1-(3-methylphenyl)-1H-pyra-
2
H, NH , D O exchangeable), 6.68 (s, 1H, pyrazole H-4),
2 2
13
.05–7.84 (m, 17H, Ar–H) ppm; C NMR (DMSO-d6):
zol-3-yl]aniline (3k, C H N O)
2
9
25
3
7
Yield 84 %; m.p.: 200–202 °C; IR (KBr): vꢀ = 3560, 3355
d = 53.2 (OCH ), 68.4 (OCH ), 115.7 (pyrazole C ),
-
1
1
NH), 3037 (CH), 1593 (C=N) cm ; H NMR (DMSO-
3
2
4
(
1
1
1
18.2, 120.4, 128.2, 128.8, 129.2, 129.4, 129.7, 130.1,
31.3, 131.6, 132.2, 132.8, 132.8, 136.3, 143.5, 144.0,
61.7 (pyrazole C ), 163.9 (pyrazole C ) ppm.
d ): d = 2.29 (s, 3H, CH ), 5.11 (s, 2H, OCH ), 5.20 (s,
6
3
2
2
7
H, NH , D O exchangeable), 6.75 (s, 1H, pyrazole H-4),
2 2
1
.05–7.85 (m, 17H, Ar–H) ppm; C NMR (DMSO-d6):
3
5
3
d = 16.1 (CH ), 68.4 (OCH ), 115.9 (pyrazole C ), 118.2,
General method for the synthesis of N-[4-[N-[4-[5-[4-
(benzyloxy)phenyl]-1-(substituted phenyl)-1H-pyrazol-3-
yl]phenyl]sulfamoyl]phenyl]acetamide 4a–4o
3
2
4
1
1
1
20.7, 128.2, 128.9, 129.0, 129.4, 130.0, 130.2, 131.1,
31.8, 132.0, 132.2, 132.4, 132.7, 132.8, 136.2, 143.7,
44.1, 161.5 (pyrazole C ), 163.6 (pyrazole C ) ppm.
Equimolar quantities of 3a–3o and p-acetamidobenzene-
sulfonyl chloride (5 mmol each) were stirred at room
temperature for 6–9 h in the presence of pyridine (5 mmol)
5
3
4
-[5-[4-(Benzyloxy)phenyl]-1-(4-methylphenyl)-1H-pyra-
zol-3-yl]aniline (3l, C H N O)
2
9
25
3
3
in 30 cm of THF. The mixture was then poured into
Yield 87 %; m.p.: 178–179 °C; IR (KBr): vꢀ = 3520, 3370
crushed ice and the precipitate obtained was filtered, dried,
and crystallized from ethanol.
-1 1
NH), 3032 (CH), 1590 (C=N) cm ; H NMR (DMSO-
(
1
23

Synthesis and pharmacological evaluation of pyrazole derivatives containing sulfonamide moiety
-
1 1
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(2-chlorophenyl)-
H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acetamide
4a, C H ClN O S)
788 (C–Cl) cm ; H NMR (DMSO-d ): d = 2.73 (s, 3H,
6
1
CH ), 5.21 (s, 2H, OCH ), 7.08 (s, 1H, pyrazole H-4),
3
2
(
7.23–8.10 (m, 20H, Ar–H), 8.45 (s, 1H, CONH, D O
2
3
6
29
4
4
Yield 78 %; m.p.: 193–194 °C; IR (KBr): vꢀ = 3332, 3321
exchangeable), 10.30 (s, 1H, SO NH, D O exchangeable)
2
2
1
3
(
NH), 3031 (CH), 1640 (C=O), 1610 (C=N), 1184 (SO ),
1 1
ppm;
(OCH ), 107.3 (pyrazole C ), 113.2, 114.1, 114.6, 115.3,
C NMR (DMSO-d ): d = 31.5 (CH ), 71.3
2
6
3
-
7
43 (C–Cl) cm ; H NMR (DMSO-d ): d = 2.76 (s, 3H,
6
2
4
CH ), 5.15 (s, 2H, OCH ), 7.05 (s, 1H, pyrazole H-4),
3
115.7, 118.7, 119.6, 120.1, 123.8, 125.9, 127.5, 128.0,
128.5, 129.4, 130.8, 131.9, 133.6, 134.1, 136.2, 137.3,
139.7, 142.4, 152.3 (pyrazole C ), 160.0 (pyrazole C ),
2
7
.07–8.38 (m, 21H, Ar–H), 8.40 (s, 1H, CONH, D O
2
exchangeable), 10.26 (s, 1H, SO NH, D O exchangeable)
2
2
5
3
?
1
3
ppm;
C NMR (DMSO-d ): d = 29.8 (CH ), 71.4
171.8 (CONH) ppm; MS: m/z = 684 ([M ? 1] ),
?
685([M ? 2] ).
6
3
(
OCH ), 105.3 (pyrazole C ), 113.7, 114.0, 114.8, 115.3,
2
4
1
1
1
1
18.7, 119.8, 121.2, 122.3, 123.5, 125.1, 128.4, 128.6,
28.7, 128.9, 130.1, 130.4, 134.7, 136.4, 136.9, 138.2,
44.6, 151.5, 159.0 (pyrazole C ), 160.8 (pyrazole C ),
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(4-fluorophenyl)-
1H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acetamide
5
3
(
4e, C H FN O S)
36 29
?
4 4
71.8 (CONH) ppm; MS: m/z = 650 ([M ? 1] ), 651
Yield 77 %; m.p.: 226–227 °C; IR (KBr): vꢀ = 3340, 3221
NH), 3025 (CH), 1669 (C=O), 1613 (C=N), 1192 (SO )
?
(
[M ? 2] ).
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(3-chlorophenyl)-
H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acetamide
4b, C H ClN O S)
(
2
-
1 1
cm ; H NMR (DMSO-d ): d = 2.92 (s, 3H, CH ), 5.21
6
3
1
(s, 2H, OCH ), 7.06 (s, 1H, pyrazole H-4), 7.11–7.86 (m,
2
(
21H, Ar–H), 8.51 (s, 1H, CONH, D O exchangeable),
2
3
6
29
4
4
1
3
Yield 71 %; m.p.: 185–186 °C; IR (KBr): vꢀ = 3328, 3217
10.24 (s, 1H, SO NH, D O exchangeable) ppm; C NMR
2 2
(
NH), 3037 (CH), 1630 (C=O), 1612 (C=N), 1191 (SO ), 792
1 1
(DMSO-d ): d = 31.5 (CH ), 72.1 (OCH ), 105.1 (pyra-
2
6
3
2
-
(
C–Cl) cm ; H NMR (DMSO-d ): d = 2.89 (s, 3H, CH ),
zole C ), 113.9, 114.3, 114.9, 115.5, 119.8, 120.4, 123.2,
4
6
3
5
.18 (s, 2H, OCH ), 7.13 (s, 1H, pyrazole H-4), 7.29–8.14
2
125.5, 127.7, 128.1, 128.9, 129.1, 130.2, 131.6, 133.4,
134.2, 135.5, 137.8, 139.2, 140.1, 152.6 (pyrazole C5),
(
m, 21H, Ar–H), 8.42 (s, 1H, CONH, D O exchangeable),
2
1
3
0.18 (s, 1H, SO NH, D O exchangeable) ppm; C NMR
1
160.7 (pyrazole C ), 171.4 (CONH) ppm; MS: m/z = 633
3
?
([M ? 1] ).
2
2
(
DMSO-d ): d = 29.3 (CH ), 71.6 (OCH ), 105.2 (pyrazole
6
3
2
C ), 113.6, 114.2, 114.6, 115.1, 118.6, 118.9, 120.8, 122.5,
4
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(4-fluoro-3-chlor-
ophenyl)-1H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acet-
amide (4f, C H ClFN O S)
1
1
23.7, 126.7, 127.3, 128.2, 128.5, 128.8, 130.1, 130.4, 133.4,
36.7, 137.5, 138.7, 142.3, 152.1, 158.8 (pyrazole C ), 159.6
5
3
6
28
4 4
(
(
pyrazole C ), 170.2 (CONH) ppm; MS: m/z = 650
3
Yield 74 %; m.p.: 224–225 °C; IR (KBr): vꢀ = 3343, 3218
NH), 3015 (CH), 1642 (C=O), 1614 (C=N), 1187 (SO ),
?
[M ? 1] ), 651 ([M ? 2] ).
?
(
2
-
1 1
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(4-chlorophenyl)-
H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acetamide
4c, C H ClN O S)
782 (C–Cl) cm ; H NMR (DMSO-d ): d = 2.74 (s, 3H,
6
1
CH ), 5.20 (s, 2H, OCH ), 7.03 (s, 1H, pyrazole H-4),
3
2
(
7.14–7.89 (m, 20H, Ar–H), 8.83 (s, 1H, CONH, D O
2
3
6
29
4
4
Yield 70 %; m.p.: 183–184 °C; IR (KBr): vꢀ = 3337, 3220
exchangeable), 10.27 (s, 1H, SO NH, D O exchangeable)
2
2
1
3
(
NH), 3031 (CH), 1645 (C=O), 1614 (C=N), 1190 (SO ),
-
ppm;
(OCH ), 105.4 (pyrazole C ), 113.1, 113.4, 113.5, 114.0,
C NMR (DMSO-d ): d = 28.5 (CH ), 71.6
2
6
3
1 1
90 (C–Cl) cm ; H NMR (DMSO-d ): d = 2.78 (s, 3H,
7
6
2
4
CH ), 5.19 (s, 2H, OCH ), 7.05 (s, 1H, pyrazole H-4),
3
114.3, 115.8, 119.0, 120.6, 121.2, 125.7, 127.2, 128.9,
129.5, 129.8, 130.6, 131.1, 133.3, 134.5, 135.8, 137.2,
139.2, 140.5, 152.6 (pyrazole C ), 160.2 (pyrazole C ),
2
7
.11–7.83 (m, 21H, Ar–H), 8.90 (s, 1H, CONH, D O
2
exchangeable), 10.22 (s, 1H, SO NH, D O exchangeable)
2
2
5
3
1
3
?
ppm;
C NMR (DMSO-d ): d = 32.1 (CH ), 71.9
172.4 (CONH) ppm; MS: m/z = 668 ([M ? 1] ), 669
?
([M ? 2] ).
6
3
(
OCH ), 105.1 (pyrazole C ), 113.4, 114.1, 114.8, 115.5,
2
4
1
1
1
16.7, 119.5, 120.5, 122.3, 123.5, 125.7, 127.7, 128.7,
28.9, 129.6, 130.3, 130.8, 133.8, 136.1, 137.4, 139.0,
42.5 (pyrazole C ), 160.7 (pyrazole C ), 171.8 (CONH)
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(4-bromophenyl)-
1H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acetamide
5
3
(
4g, C H BrN O S)
36 29
?
?
4 4
ppm; MS: m/z = 650 ([M ? 1] ), 651 ([M ? 2] ).
Yield 72 %; m.p.: 220–222 °C; IR (KBr): vꢀ = 3345, 3320
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(3,4-dichlor-
ophenyl)-1H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acet-
amide (4d, C H Cl N O S)
(NH), 3018 (CH), 1645 (C=O), 1608 (C=N), 1183 (SO )
-
2
1 1
cm ; H NMR (DMSO-d ): d = 2.76 (s, 3H, CH ), 5.21
6
3
(s, 2H, OCH ), 7.06 (s, 1H, pyrazole H-4), 7.11–7.89 (m,
2
3
6
28
2
4
4
Yield 77 %; m.p.: 220–222 °C; IR (KBr): vꢀ = 3342, 3219
NH), 3028 (CH), 1665 (C=O), 1609 (C=N), 1187 (SO2),
21H, Ar–H), 8.72 (s, 1H, CONH, D O exchangeable),
2
1
3
(
10.19 (s, 1H, SO NH, D O exchangeable) ppm; C NMR
2 2
123

K. Somakala et al.
(
DMSO-d ): d = 32.6 (CH ), 72.1 (OCH ), 105.3 (pyra-
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(3-methylphenyl)-
1H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acetamide
(4k, C H N O S)
6
3
2
zole C ), 113.7, 114.3, 114.8, 115.5, 119.6, 120.2, 121.2,
4
1
1
1
25.7, 127.1, 128.5, 129.0, 129.4, 130.3, 131.2, 133.9,
3
7 32 4 4
34.3, 135.5, 137.3, 138.6, 142.5, 152.5 (pyrazole C5),
Yield 73 %; m.p.: 155–156 °C; IR (KBr): vꢀ = 3337, 3225
60.8 (pyrazole C ), 172.6 (CONH) ppm; MS: m/z = 694
3
(NH), 3021 (CH), 1640 (C=O), 1611 (C=N), 1189 (SO )
-1 1
2
?
(
[M ? 1] ).
cm ; H NMR (DMSO-d ): d = 2.17 (s, 3H, CH ), 3.12
6
3
(
s, 3H, CH ), 5.20 (s, 2H, OCH ), 7.07 (s, 1H, pyrazole
3 2
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(4-nitrophenyl)-
H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acetamide
4h, C H N O S)
H-4), 7.12–7.89 (m, 21H, Ar–H), 8.71 (s, 1H, CONH, D O
2
1
exchangeable), 10.31 (s, 1H, SO NH, D O exchangeable)
2
13
ppm; C NMR (DMSO-d ): d = 17.1 (CH ), 32.9 (CH ),
7
1
1
1
2
(
3
6
29
5
6
6
3
3
Yield 71 %; m.p.: 230–232 °C; IR (KBr): vꢀ = 3338, 3221
NH), 3014 (CH), 1642 (C=O), 1610 (C=N), 1190 (SO )
1.5 (OCH ), 109.3 (pyrazole C ), 112.3, 114.0, 114.1,
2 4
(
2
14.4, 115.3, 115.8, 119.1, 122.0, 124.4, 126.7, 127.6,
28.2, 129.1, 129.6, 130.4, 133.5, 134.6, 135.2, 135.8,
-
1 1
cm ; H NMR (DMSO-d ): d = 2.94 (s, 3H, CH ), 5.22
6
3
(
s, 2H, OCH ), 7.04 (s, 1H, pyrazole H-4), 7.16–7.83 (m,
2
38.2, 138.9, 140.6, 155.7 (pyrazole C ), 160.7 (pyrazole
5
2
1H, Ar–H), 8.87 (s, 1H, CONH, D O exchangeable),
2
?
C ), 171.3 (CONH) ppm; MS: m/z = 629 ([M ? 1] ).
3
1
0.29 (s, 1H, SO NH, D O exchangeable) ppm; C NMR
3
1
2
2
(
DMSO-d ): d = 32.3 (CH ), 71.8 (OCH ), 109.2 (pyra-
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(4-methylphenyl)-
1H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acetamide
(4l, C H N O S)
6
3
2
zole C ), 113.6, 114.2, 114.8, 115.1, 119.4, 120.8, 124.7,
4
1
1
1
25.1, 127.8, 128.3, 129.4, 129.9, 130.1, 133.2, 133.6,
3
7 32 4 4
34.2, 135.7, 137.8, 138.9, 142.2, 155.0 (pyrazole C5),
Yield 73 %; m.p.: 159–160 °C; IR (KBr): vꢀ = 3341, 3220
60.4 (pyrazole C ), 171.8 (CONH) ppm; MS: m/z = 660
3
(NH), 3014 (CH), 1637 (C=O), 1614 (C=N), 1190 (SO )
-1 1
2
?
(
[M ? 1] ).
cm ; H NMR (DMSO-d ): d = 2.04 (s, 3H, CH ), 2.73
6
3
(
s, 3H, CH ), 5.18 (s, 2H, OCH ), 7.03 (s, 1H, pyrazole
3 2
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(2,4-dinitro-
phenyl)-1H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acet-
amide (4i, C H N O S)
H-4), 7.15–7.87 (m, 21H, Ar–H), 8.73 (s, 1H, CONH, D O
2
exchangeable), 10.20 (s, 1H, SO NH, D O exchangeable)
2
13
ppm; C NMR (DMSO-d ): d = 17.4 (CH ), 32.9 (CH ),
7
1
1
1
2
3
6
28
6
8
6
3
3
Yield 71 %; m.p.: 256–257 °C; IR (KBr): vꢀ = 3342, 3220
NH), 3017 (CH), 1647 (C=O), 1607 (C=N), 1188 (SO )
1.1 (OCH ), 110.3 (pyrazole C ), 112.9, 113.8, 114.7,
2 4
(
2
15.2, 119.6, 122.8, 124.1, 126.3, 127.5, 128.2, 129.0,
29.8, 130.2, 133.9, 134.5, 134.7, 135.2, 138.6, 138.9,
40.1, 155.3 (pyrazole C ), 160.1 (pyrazole C ), 172.3
-
1 1
cm ; H NMR (DMSO-d ): d = 2.76 (s, 3H, CH ), 5.19
6
3
(
s, 2H, OCH ), 7.08 (s, 1H, pyrazole H-4), 7.14–7.89 (m,
2
5
3
2
0H, Ar–H), 8.76 (s, 1H, CONH, D O exchangeable),
2
?
(
CONH) ppm; MS: m/z = 629 ([M ? 1] ).
1
0.22 (s, 1H, SO NH, D O exchangeable) ppm; C NMR
3
1
2
2
(
DMSO-d ): d = 31.3 (CH ), 72.8 (OCH ), 105.7 (pyra-
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(2,6-dimethylphe-
nyl)-1H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acetamide
(4m, C H N O S)
6
3
2
zole C ), 112.1, 113.8, 114.2, 114.7, 115.4, 118.7, 119.6,
4
1
1
20.2, 124.3, 125.7, 127.2, 128.1, 129.3, 129.7, 130.5,
33.6, 134.1, 135.2, 135.6, 137.1, 138.2, 141.1, 155.8
3
8 34 4 4
Yield 70 %; m.p.: 262–264 °C; IR (KBr): vꢀ = 3339, 3221
(
pyrazole C ), 160.7 (pyrazole C ), 171.2 (CONH) ppm;
5
(NH), 3013 (CH), 1642 (C=O), 1612 (C=N), 1186 (SO )
-1 1
3
2
?
MS: m/z = 705 ([M ? 1] ).
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(2-methylphenyl)-
H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acetamide
4j, C H N O S)
cm ; H NMR (DMSO-d ): d = 2.18 (s, 6H, 2 CH ), 3.01
6
3
(s, 3H, CH ), 5.19 (s, 2H, OCH ), 7.08 (s, 1H, pyrazole
3 2
H-4), 7.13–7.83 (m, 20H, Ar–H), 8.33 (s, 1H, CONH, D O
2
1
exchangeable), 10.18 (s, 1H, SO NH, D O exchangeable)
2
2
(
3
7
32
4
4
13
ppm; C NMR (DMSO-d ): d = 17.7 (2 CH ), 32.6
6
3
Yield 72 %; m.p.: 150–152 °C; IR (KBr): vꢀ = 3343, 3323
NH), 3019 (CH), 1642 (C=O), 1616 (C=N), 1191 (SO )
(CH ), 71.3 (OCH ), 110.3 (pyrazole C ), 113.2, 114.4,
3 2 4
(
2
1
1
1
1
14.9, 115.4, 119.7, 122.7, 124.3, 127.3, 127.6, 128.8,
29.1, 129.5, 130.9, 133.9, 134.7, 134.9, 135.0, 138.8,
38.9, 140.3, 155.5 (pyrazole C ), 160.7 (pyrazole C ),
-
1 1
cm ; H NMR (DMSO-d ): d = 2.28 (s, 3H, CH ), 2.98
6
3
(
s, 3H, CH ), 5.17 (s, 2H, OCH ), 7.05 (s, 1H, pyrazole
3 2
5
3
H-4), 7.18–7.82 (m, 21H, Ar–H), 8.77 (s, 1H, CONH, D O
2
?
71.3 (CONH) ppm; MS: m/z = 643 ([M ? 1] ).
exchangeable), 10.29 (s, 1H, SO NH, D O exchangeable)
2
2
1
3
ppm; C NMR (DMSO-d ): d = 18.2 (CH ), 32.3 (CH ),
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(2-methoxy-
phenyl)-1H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acet-
amide (4n, C H N O S)
6
3
3
7
1
1
1
1.8 (OCH ), 105.7 (pyrazole C ), 112.4, 113.2, 113.6,
2 4
14.1, 114.5, 115.6, 119.2, 120.4, 124.8, 126.9, 127.8,
28.4, 129.3, 129.8, 130.1, 134.6, 134.9, 135.1, 135.8,
3
7 32 4 5
Yield 74 %; m.p.: 285–286 °C; IR (KBr): vꢀ = 3342, 3217
38.3, 138.5, 141.6, 155.2 (pyrazole C ), 160.3 (pyrazole
5
(NH), 3017 (CH), 1653 (C=O), 1610 (C=N), 1191 (SO )
-1 1
2
?
C ), 169.9 (CONH) ppm; MS: m/z = 629 ([M ? 1] ).
3
cm ; H NMR (DMSO-d ): d = 2.89 (s, 3H, CH ), 3.71 (s,
6
3
1
23

Synthesis and pharmacological evaluation of pyrazole derivatives containing sulfonamide moiety
3
H, OCH ), 5.21 (s, 2H, OCH ), 7.06 (s, 1H, pyrazole H-4),
3
potential as described earlier [25]. Stock solutions of dif-
ferent compounds (1 mM) were mixed with DPPH
2
7
.14–7.89 (m, 21H, Ar–H), 8.32 (s, 1H, CONH, D O
2
3
3
exchangeable), 10.26 (s, 1H, SO NH, D O exchangeable)
2
methanol solution (0.5 cm , 0.3 mM) in 3 cm of total
reaction mixture and allowed to react at room temperature.
After 30 min, absorbance was measured at 520 nm and
converted to % antioxidant activity. For a comparative
study BHA was used as the standard. The percentage
inhibition activity was calculated by using the formula:
2
1
3
ppm; C NMR (DMSO-d ): d = 32.0 (CH ), 56.1 (OCH ),
6
3
3
7
1
1
1
1.3 (OCH ), 110.1 (pyrazole C ), 113.4, 113.7, 114.2,
2 4
14.9, 115.4, 119.7, 122.5, 124.0, 127.4, 127.6, 128.1, 129.5,
29.8, 130.4, 133.1, 133.3, 134.6, 134.9, 135.8, 138.3, 138.5,
40.2, 155.0 (pyrazole C ), 160.1 (pyrazole C ), 171.6
5
3
?
(
CONH) ppm; MS: m/z = 647 ([M ? 1] ).
%
activity ¼ ½1 À ðA =A ފ  100
t
c
N-[4-[N-[4-[5-[4-(Benzyloxy)phenyl]-1-(4-methoxy-
phenyl)-1H-pyrazol-3-yl]phenyl]sulfamoyl]phenyl]acet-
amide (4o, C H N O S)
where A = absorbance of test compound and A absor-
t
c
bance of control compound.
3
7 32 4 5
Yield 74 %; m.p.: 286–287 °C; IR (KBr): vꢀ = 3337, 3215
NH), 3010 (CH), 1653 (C=O), 1615 (C=N), 1186 (SO )
In vivo studies
(
2
-
1 1
cm ; H NMR (DMSO-d ): d = 2.72 (s, 3H, CH ), 3.76
6
3
Twelve-week-old adult male/female Wistar rats
150–200 g) and Balb/c mice (20–30 g) used in the present
(
s, 3H, OCH ), 5.21 (s, 2H, OCH ), 7.08 (s, 1H, pyrazole
3 2
(
H-4), 7.14–7.92 (m, 21H, Ar–H), 8.34 (s, 1H, CONH, D O
2
study were housed and kept in accordance with the Ham-
dard University Animal Care Unit, which applies the
guidelines and rules laid down by Committee for the pur-
pose of control and supervision of Experiments on Animals
exchangeable), 10.20 (s, 1H, SO NH, D O exchangeable)
2
2
1
3
ppm;
C NMR (DMSO-d ): d = 32.6 (CH ), 57.2
6
3
(
OCH ), 71.6 (OCH ), 110.7 (pyrazole C ), 113.2, 113.2,
3 2 4
1
1
1
1
14.6, 115.1, 119.7, 120.7, 122.6, 124.5, 127.3, 127.4,
28.9, 129.0, 129.6, 130.1, 134.5, 134.6, 135.8, 138.3,
38.5, 140.8, 153.7 (pyrazole C ), 160.6 (pyrazole C ),
(
CPCSEA), Ministry of Social Justice and Empowerment,
Government of India. Animals were housed in a ventilated
room at 25 ± 2 °C under a 12 h light/dark cycle in large
spacious polypropylene cages. Animals were allowed to
acclimatize for 1 week before the study and had free access
to standard laboratory feed and water ad libitum.
5
3
?
72.4 (CONH) ppm; MS: m/z = 647 ([M ? 1] ).
Anti-inflammatory screening
The synthesized compounds were screened for anti-inflam-
matory activity by the inhibition of albumin denaturation
technique [24]. Diclofenac sodium (standard drug) and test
compounds were dissolved in minimum amount of DMF and
diluted with phosphate buffer saline (pH 7.4) in such a way
that concentration of DMF in all solutions was less than
Anti-inflammatory activity
The synthesized compounds were evaluated for their anti-
inflammatory activity using carrageenan induced rat hind
paw edema method [26]. The animals were randomly
allocated into groups of six animals each and fasted for
3
3
2
.5 %. Test solutions (1 cm , 100 lg/cm ) were mixed with
24 h before the experiment with free access to water.
3
cm of 1 % albumin solution in phosphate buffer saline
1
Control group received only 0.5 % CMC solution. Stan-
dard, diclofenac sodium was administered orally at a dose
of 10 mg/kg. The test compounds were administered orally
at an equimolar oral dose relative to 10 mg/kg diclofenac
and incubated at 27 ± 1 °C in an incubator for 15 min.
Denaturation was induced by keeping the reaction mixture at
6
0 ± 1 °C in a water bath for 10 min. After cooling, the
turbidity was measured at 660 nm with UV–Vis spec-
trophotometer. Percentage inhibition of denaturation was
then calculated from control where no drug was added. Each
experiment was done in triplicate and average taken. The
percentage of inhibition was calculated using the formula:
3
sodium. 0.1 cm of 1 % carrageenan solution in saline was
injected subcutaneously into the sub plantar region of the
right hind paw of each rat, 1 h after the administration of
the test compounds and standard drug. The right hind paw
volume was measured before and after 3 and 4 h of car-
rageenan treatment by means of a plethysmometer. The
percent edema inhibition was calculated from the mean
effect in the control and treated animals according to the
following equation:
%
inhibition of denaturation ¼ ½ðVt=VcÞ À 1Š  100
where V = mean absorption of test compound and
t
Vc = mean absorption of control.
%
edema inhibition ¼ ðV À V =V Þ Â 100
c
t
c
Antioxidant assay
where V represents the mean increase in paw volume in
t
0
All the compounds were evaluated in vitro for their 2,2 -
rats treated with test compounds and V
represents the
c
diphenyl-1-picrylhydrazyl (DPPH) radical scavenging
mean increase in paw volume in control group of rats. For
123

K. Somakala et al.
the ulcerogenic activity the same group of rats which were
used for anti-inflammatory activity were used after a
washout period of 15 days.
Determination of the inhibition of LPS-induced
TNF-a production
TNF-a production inhibition in mice (Balb/c) weighing
20–30 g was carried out by the already reported procedure
Ulcerogenic activity
[
29, 30]. Mice were deprived of food overnight, but not
The ulcerogenic effect of five active compounds and
diclofenac sodium was evaluated by the reported method
water. The next day, these mice were orally administered
with test compounds at an equimolar oral dose relative to
30 mg/kg SB 203580 inhibitor suspended in 0.5 % CMC
[
27]. The rats were allocated into different groups con-
3
sisting of six animals in each group. Ulcerogenic activity
was evaluated after oral administration of the test com-
pounds at an equimolar dose relative to 30 mg/kg
diclofenac sodium. Control group received only 0.5 %
carboxymethylcellulose (CMC) solution. Food but not
water was stopped 24 h before administration of the test
compounds. After the drug treatment, the rats were fed
with normal diet for 17 h and then sacrificed. The stomach
was removed and opened along the greater curvature,
washed with distilled water, and cleaned gently by dipping
in normal saline. The mucosal damage was examined by
means of a magnifying glass. For each stomach the
mucosal damage was assessed according to the following
scoring system: 0.5: redness, 1.0: spot ulcers, 1.5: hemor-
rhagic streaks, 2.0: ulcers [3 but B5, 3.0: ulcers [5. The
mean score of each treated group minus the mean score of
control group was regarded as severity index of gastric
mucosal damage.
aqueous solution at 10 cm /kg. The groups of normal and
control mice were administered only with 0.5 % CMC
3
aqueous solution at 10 cm /kg. Thirty minutes later, LPS
solution dissolved in physiological saline was intra-
3
venously injected at 0.45 mg/10 cm /kg except for the
group of normal mice, which were injected with physio-
3
logical saline at 10 cm /kg. One hour later, blood samples
were taken from the mice from the vena cava inferior into a
syringe containing heparin sodium. After centrifugation of
the blood samples at 13,230g for 3 min at 4 °C, plasma
samples were taken immediately and frozen at -20 °C
until measurement of the concentration of TNF-a. The
concentrations of TNF-a in the plasma samples were
measured with commercially available ELISA kit. The
percent inhibition of TNF-a production was obtained by
the following equation: Percent inhibition = [1 - (con-
centration of TNF-a in plasma sample of mice
administrated test compounds - mean concentration of
TNF-a in the plasma samples of normal mice)/(concen-
tration of TNF-a in the plasma samples in control
mice - mean concentration of TNF-a in the plasma sam-
ples of normal mice)] 9 100.
Lipid peroxidation
Lipid peroxidation in the gastric mucosa was determined
according to the method of Ohkawa et al. [28] method.
After screening the stomach for ulcers the gastric mucosa
of glandular portion was scrapped with the help of two
glass slides. Gastric mucosa was weighed (100 mg) and
Molecular docking study
For the purpose of assessment of docking of ligands to
protein active sites for estimation of binding affinities of
docked compounds, an advanced molecular docking pro-
gramme GLIDE (Schrodinger Inc., USA) version 9.8 was
used. GLIDE, the Grid-based ligand docking with ener-
getics algorithm approximates a systematic search of
positions, orientations and conformations and eliminates
unwanted conformations using scoring in the enzyme
pocket via a series of hierarchical filters. Finally the con-
formations were refined via Monte Carlo sampling of pose
conformation. Glide provides three different levels of
docking precisions viz. High Throughput Virtual Screen-
ing, HTVS; Standard precision, SP and Extra precision,
XP. We carried out our calculations using XP docking
mode as the tool is designed for better refinement in
ligands. X-ray crystal structure of p38a MAP kinase was
3
homogenized in mortar and pestle with 1.8 cm ice-cold
1
0
.15 % KCl solution. The homogenate supplemented with
3
.2 cm of 8.1 % sodium dodecyl sulfate (SDS), 1.5 cm of
3
3
acetate buffer, and 1.5 cm of 0.8 % thiobarbituric acid.
The mixture was then incubated at 95 °C for 60 min on
boiling water bath. The reaction mixture was kept at room
temperature for some time and then extracted with a
3
mixture of n-butanol: pyridine (15:1, v/v; 5 cm ) by
shaking vigorously for 1 min and then keeping in ice for
2
1
min. The organic layer was centrifuged at 3000 rpm for
0 min. Organic layer was separated out and absorbance
measured at 532 nm on UV spectrophotometer. Results
were expressed as nmol MDA/100 mg tissue using
5
extinction coefficient 1.56 9 10 cm
-1
-1
M . Concentra-
9
tion = (absorbance 9 volume 9 10 )/(1.56 9 10 ).
8
1
23

Synthesis and pharmacological evaluation of pyrazole derivatives containing sulfonamide moiety
˚
taken from PDB entry 3D83 having resolution of 1.90 A.
12. Bekhit AA, Fahmy HTY, Rostom SAF, Bekhit AE (2010) Eur J
Med Chem 45:6027
For docking studies protein structures were prepared using
protein preparation wizard in Maestro 9.8. Protein prepa-
ration was carried out in two steps viz. preparation and
refinement. Chemical correctness was ensured and water
molecules in crystal structure were deleted and hydrogen
atoms were added at missing positions and bond order for
crystal ligand and protein was adjusted and minimized up
1
3. Adhikari A, Kalluraya B, Sujith KV, Gouthamchandr K, Jairamc
R, Mahmood R, Sankolli R (2012) Eur J Med Chem 55:467
14. Vijesh AM, Isloor AM, Shetty P, Sundershan S, Fun HK (2013)
Eur J Med Chem 62:410
1
5. Ashour HMA, El-Ashmawy IM, Bayad AE (2015) Monatsh
Chem. doi:10.1007/s00706-015-1549-x
6. Singh D, Bansal GE (2004) J Chem 1:164
1
17. Penning TD, Talley JJ, Bertenshaw SR, Carter JS, Collins PW,
Docter S, Graneto MJ, Lee LF, Malecha JW, Miyashiro JM,
Rogers RS, Rogier DJ, Yu SS, Anderson GD, Burton EG, Cog-
burn JN, Gregory SA, Koboldt CM, Perkins WE, Seibert K,
Veenhuizen AW, Zhang YY, Isakson PC (1997) J Med Chem
40:1347
˚
to 0.30 A RMSD. Ligprep 2.2 module in maestro 9.8 build
panel was used for ligand preparation which produced low
energy conformations of ligand using OPLS 2005 force
field [31].
1
8. Talley JJ, Brown DL, Carter JS, Graneto MJ, Koboldt CM,
Masferrer JL, Perkins WE, Rogers RS, Shaffer AF, Zhang YY,
Zweifel BS, Seibert K (2000) J Med Chem 43:775
Acknowledgments The authors are grateful to Jamia Hamdard for
providing infrastructure and other facilities. The author (Kanagasabai
Somakala) is thankful to Department of Science and Technology
Women Scientists Scheme-A (File No.: SR/WOS-A/CS-84/2013),
New Delhi for providing financial assistance.
19. Amir M, Kumar H, Javed SA (2008) Eur J Med Chem 43:2056
20. Amir M, Saifullah K, Akhter W (2011) J Enz Inhib Med Chem
26:141
2
2
1. Amir M, Somakala K, Ali S (2013) Min Rev Med Chem 13:2082
2. Amir M, Akhter MW, Alam O (2015) Monatsh Chem. doi:10.
1
007/s00706-015-1557-x
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